Diy GF Torani LiBRARY Digitized by the Internet Archive in 2010 with funding from University of Toronto http://www. archive.org/details/annalsoftropical15live ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY — avioiaal .aoioaT. 10" YOO1O TIARA EA @ (THE UNIVERSITY OF LIVERPOOL g he , ANNALS we ee TropicaL MEDICINE AND PARASITOLOGY ISSUED BY THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE Edited by Prorrssor J. W. W. STEPHENS, M.D.Cantab., F.R.S. Prorrssor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.ALS. Proressor WARRINGTON YORKE, M.D. Prorrssor B. BLACKLOCK, M.D. VOLUME XV (April 27, 1921, to December 30, 1921) af 4 ‘ With Frontispiece, thirty-one plates, one hundred and twenty-seven Jigures in text, five maps, and four charts LIVERPOOL : AT THE UNIVERSITY PRESS, 57 ASHTON STREET. 4 = F a an ee, ‘ . AMA amioraa We akg ti avioigdM JA0ITOA T Weg 26eiae ali —. , i ~ sr z ia se FF AR iain, CTA 2 tH} WE ae * RE Jott oe ae he Bee ith Pg : A Set Peed? AHOY fe secs sos : JX -BMUIOV isQ% O€ mdmessG ah whores i X) Pe Vn it : i v? SM fe SW bel - ai) ‘ awit fy Pek ta ge! . % a , i 2 Sogn s ee DM RAY ARAN. TE est mH CONTENTS No. I. April 27, 1921 Aritmanp, Dorotny. PAGE Liverpool School of ‘Tropical Medicine. Scientific Record... at 7% I Brent, A, On the ‘ Arneth Count’ in Hookworm-Infected White Children in North Queensland 5c) Sees (ebey aa epee egress ited in oA AD Macrtz, J. W. S.; and Incram, A. Bronchomoniliasis Complicating Pulmonary ‘Tuberculosis in a Native of the Gold Coast, West Africa ee Rae eae cis ie tis sa 53 Bracktoek, B. Notes on a Case of Indigenous Infection with P. falciparum... = 3. 59 Bracktock, B.; and Carrer, H. F. Observations on Mosquitoes in the Isle of Man ... sas ae oe Bee 73 Hus, G. F. Notes on Some Unusual Breeding-places of Stegomyia fasciata, Fabr., in JACEE ae he cee sete eres Boas ee thay nice ant-wjat) OF Hitt, G. F. Musca domestica, Linn., as a ‘ Bush Fly’ in Australia’... AY oe oo 93 Newsteap, R.; and Evans, Atwen M. New Tsetse-Flies (Glossina) from the Belgian Congo ... FE See ais 95 NEWSTEAD, R.; and Sinton, J. A. On a Collection of Pappataci Flies (Ph/ebotomus) from India... a spty A10F CONTENTS No. 2. July 16, 1921 Sinton, J. A. PAGE Notes on Oriental Sore in Russian Turkestan and the Results of Treatment with Injections of Tartar Emetic Solution ... a5 50 a She in 7) Barret, H. P. A Method for the Cultivation of Blastocystis Fiat nat ae La ae eS Scott, Henry Haroitp The Incidence of Intestinal Parasites, especially with regard to the Protozoa, amongst Symptomless Carriers in Jamaica ... ae ace Sc veg EY, Scotr, Henry Haroip A Study of the Sizes of Entamoeba histolytica Cysts amongst Symptomless Carriers in Jamaica Ss say one 505 o54 500 th rem CE Scorr, Henry Haroip A Study of the Sizes of Entamoeba coli Cysts amongst Symptomless Carriers in Jamaica ... 596 a0 Sn 508 ae ae 400 ae on 2AQ SouTuwett, T. Cestodes from Indian Poultry ... S00 ee 500 40 300 ee aes Soutuwet., T. Cestodes from African Rats... ... 260 00 53 ue a0 ae) wr LO. Soutnwe tt, T. A New Species of Cestoda from a Cormorant... ie ate oes acs G9 StepuHens, J. W. W.; and Apter, S. A Case of Suspected Leprosy ... ... sc 500 36 00 = ian ZS vi CONTENTS No. 3. September 30, 1921 Carter, Henry F.; Incram, A.; and Macriz, J. W. S. Observations on the Ceratopogonine Midges of the Gold Coast with Descriptions of New Species, Part IV Scorr, Henry Harotp. The Prevalence and Character of ‘Tuberculosis in Hongkong. Part I Scott, Henry Haro.p. The Prevalence and Character of Tuberculosis in Hongkong. Part II Kennan, R. H. Multiple Aneurysms in a Child Yorke, Warrincton ; and Soutuwett, T. Lappeted Anoplocephala in Horses ... Gorvon, R. Montcomery; and Younes, C. J. The Feeding Habits of Stegomyza calopus, Meigen Yorke, Warrincton ; and Apter, S. Note on a Case of Leprosy ... Macriz, J. W. S. Notes on some Fungal Infections in West Africa ... Macriz, J. W. S.; and Incran, A. A Fungus of the genus Nocardia Cultivated from Heart Blood Newsteap, R.; and Evans, Atwen M. Report on Rat-Flea Investigation ... Younc, C. J. Natural Enemies of Stegomyta calopus, Meigen PAGE 227 245 249 265 269 271 283 287 301 CONTENTS No. 4. December 30, 1921 Incram, A.; and Macrig, J. W. S. West African Ceratopogoninae Macriz, J. W. S. The Effect of Saline Solutions and Sea-water on Stegomyia fasciata Scott, Henry Haroxp. The Prevalence and Character of Tuberculosis in Hongkong. Part III Ture, J. E. W- On the Genus Cylicostomum Maptestone, P. A. Notes on Australian Cestodes. Part I Maptestong, P. A. Notes on Australian Cestodes. Part II Maptrstong, P, A. Notes on Ulcerative Granuloma MacGrecor, Matcorm E. The Structural Differences in the Ova of Anopheles maculipennis, A. bifurcatus and 4. plumbeus Apter, S. The ‘Trypanocidal Effect of Phenylglycine Amido Arsenate of Sodium on T. brucei in Rats and I. rhodestense in Mice ... Apter, 8S. Note on Bismuth as a Trypanocide... Srepuens, J. W. W. Malaria on a Venezuelan Oilfield Evans, Atwen M. Notes on Culicidae Collected in Venezuela SourHweti, T.; and Mapuestone, P. A. A Note on the Synonymy of the Genus dla ais Poe and of the Species Z. guineensis (Graham, 1908) .. Davey, J. B.; and Newsreap, R. Mosquitoes and other Blood- ene aaah sae of the Up Pper Shiri poe Nyasaland Bracxtock, B. Breeding Places of Anopheline Mosquitoes in Freetown, Sierra Leone... Bracktock, B. Notes on an Apparatus for the Individual Breeding of Mosquitoes Yorke, WARRINGTON. The Treatment of a Case of Rhodesian emis Sickness eh the Preparation known as ‘ Bayer 205’ sae vu PAGE 313 377 381 397 493 497 413 417 427 433 435 445 455 457 463 473 479 “ re AUCTAE r . « ‘ “ . 4 id % e : ’ SA an sie ‘ 7 * : . aE - ; INDEX NEES OF AUTHORS: 5:55 0cc0cscsttsatasccnes REENERATo UNDER: Method for cultivation of 115 Nature of. 7-.225-2--eeeeeee II3 Blood- “sucking arthropods of the Upper Shiri River, Nyasaland... .. 457 Breeding of mosquitoes, Apparatus for 473 5s places of anopheline mosqui- toes in Freetown, Sierra Leone...... 463 Breinl, A. On the ‘ Arneth Count’ in hookworm-infected white chil- dren in North Queensland ......... 49 Brevicapsulatum group of Cylicostomum 401 Bronchomoniliasis complicating pul- monary tuberculosis in a native of the ‘Gold’'Goast:7ssseience sree eeeees 53, 285 Calicatum group of Cylicostomum PAGE Carter, H. F., and Blacklock, B. Observations on mosquitoes in the Esl of Mansi sc90 <0 os003~0 sree eee Carter, H. F., Ingram, A., and Macfie. Observations on the Ceratopogonine midges of the Gold Coast, with descriptions of new species.............. 177 Cellia argyrotarsis in Venezuela......... 439 Ceratophyllus fasciatus on rats at Liver- j POolbe=:..84..--b-.,053 292 5 londiniensis on rats at Liverpool, ,» irregular distribution in AUSEOOL s 2 ixekcnseen or ooee =" histolytica cysts, Sizes of, in symptomless CaLteLs....c0-0ee danger of spread by healthy CATTIEFS. «12.0.8 differentiation from E, coli... Incidence of, in symptomless carriers ...120, 124 an spread by Bee 126 Entomological research by the Tayer: pool SERB fs komt Y Roost Etorleptiomyia mediolineata.............+. 460 Eukratohelea, gen. nov. ....ccceceeeeeeees 347 ” ” vi PAGE Eukraiohelea africana, sp.n., adult ... 347 > versicolor, sp.n., adult ... 351 Euproctus-bicoronatum group of Cyli- COSLOMUM oa rcessvscecssceesevenccccsescces Evans, A. M. Notes on Culicidae collected in Venezuela ............... 5 Evans, A. M., and Newstead, R. New tsetse-flies (Glossina) from the Bel- pian Congo! 'F.. 222 iiis seis eee Evans, A. M., and Newstead, R. Report on rat-flea investigation...... Feeding habits of 8. calopus ...........- Fleas occurring on rats at Liverpool... Flies as carriers of E. Aistolytica......... Filariasis research by the Liverpool School. ......:5s..caneuee eee eee Fungal infections in West Africa Tungus of the genus Nocardia culti- vated from heart blood...............+5 Giardia intestinalis, even distribution in a stool......... Incidence of, in symptomless carriers ...120, 127 Glossina, new, from the Belgian Congo 95 >> fusca, genital armature 102 var. congolensis, 126 283 118 sees ” ” var. congolensis, var. nov., genital arma- ture? (ex eee = schwetzi, sp.n., genital arma- ” ” 102 BA 5» morphology 55 tabaniformis, genital armature Gold Coast,-Bronchomoniliasis in...53, 285 3 »» Ceratopogoninae of......... 177 Gordon, R. M., and Young, C. J. The feeding habits of Stegomyia CALOPUS. ..secscaeds+eceonsgenden eee Granuloma, ulcerative, Notes on Gyrocoelia genus, Diagnosis of Helminthiasis research by the Liver- pool!School. i. <. vss.secemeusestite eee Hemiptera from Nyasaland............... Heteroplus grandis from an Indian jungle fowl Hill, G. F. Alusea domestica as a ‘bush-fly ’? in Australia ............... Hill, G. F. Notes on some unusual breeding-places of Stegomyia fasciata in Australia..5... ated. cssnueeme eee - 93 PAGE Hongkong, Sanitary and housing con- INET TI se oe as se 239 on 219 ss Tuberculosis in ...213, 227, 381 Hookworm-infected children, Arneth ERT ER Era tenicenccns tase tens coca 49 Horses, Lappeted Anoplocephala in ... 249 Hyalomma aegyptium.........cccceeceeveees 462 Thle, J. E. W. On the genus Cy/i- RN Een en css agennec cas 397 India, Pappataci flies from. ............ 103 Indian poultry, Cestodes from ......... 161 Indigenous infection with P. falciparum 59 Ingram, A., and Macfie, J. W. S. Bronchomoniliasis complicating pul- monary tuberculosis in a native of the Gold Coast 53 Ingram, A., and Macfie, J. W. S. A fungus of the genus Nocardia culti- vated from heart blood................. 283 Ingram, A., and Macfie, J. W. S. West African Ceratopogoninae ...... 313 Ingram, A., Carter, H. F’., and Macfie, J. W.S. Observations on the Cera- topogonine midges of the Gold Coast, with descriptions of new Specter eree tect coe sec cenctc sevens sashece 177 Ingramia uniformis ........600..cs00eeeeeee 460 Intestinal parasites among symptom- less carriers in Jamaica ............... 117 Isle of Man, Mosquitoes in ............ 73 Ixodidae from Nyasaland ............... 461 Jamaica, Sizes of E. coli cysts in symp- . tomless carriers in 149 Ba » _ E. histolytica cysts in symptomless car- Miers Ul: sere, tone 133 » Symptomless carriers of in- testinal parasites in ......... 117 BMNMEMATARPLIDIIS<., 0°. 0-cc-0..skaceccesese 446 Kempia ochrosoma, sp.n., adult ......... 340 rr Ps Te ALWAS cccaen 343 Ree PUPA ies cace ts 342 Kennan, R. i. Multiple aneurysms RIMMEL sae hascccenssns secs cvte ise 245 Knapp’s obseryations on Arneth count in hookworm patients .............4. 49 Krusei group of Monilia... sets 279 Lappeted Anoplocephala in 1 horses 249 Leishmaniasis treated with tartar emetic solution.. pees 107 Leprosy, Case of suspected... Abt cessor 173 ae WWote OM #/case Of:..5.A..-0.00- 269 PAGE Leptopsylla musculi on rats at Liverpool 293 Libellulidae destructive to S. calopus... 302 Liverpool, Fleas occurring on rats at... 287 a School of Tropical Medi- cine: Scientific Record 1 Macfie, J. W. S. The effect of saline solutions and sea-water on Stegomyia U le 2 ee i ae S ARN a ee a 377 Macfie, J. W. S. Notes on some fungal infections in West Africa...... 271 Macfie, J. W. S., and Ingram, A. Bronchomoniliasis complicating pul- monary tuberculosis in a native of PHEMGOIECOASE css secceres cen ecvs 53, 285 Macfie, J. W. S., and Ingram, A. A fungus of the genus Nocardia culti- vated from heart blood................. 283 Macfie, J. W. S., and Ingram, A. West African Ceratopogoninae...... 313 Macfie, J. W. S., Carter, H. F., and Ingram, A. Observations on thé Ceratopogonine midges of the Gold Coast, with descriptions of new ROCCE ree see ee ete tes cect ewe non canes@s 177 Macfie’s observations on Arneth count in hookworm patients ................68 50 MacGregor, M. E. The structural differences in the ova of Anopheles maculipennis, A. bifurcatus, and A. UOPIL EE tee aeiccn aver nesses cet eerkowens' 417 Malaria, case of indigenous malignant RONUAL Coactccattcrcssscbesesst 59 » ona Venezuelan oilfield ...... 435 » research by the Liverpool PCHO ar auases secrets - 274. 5 from a case of tuber- culosis, 55, 274, 285 i africana, sp.n., Biochemical reaction of 277 from a case of diarrhoea... 275 Pe enterica, Biochemical reaction Of; 5. 0ckcsennessneseemeschees 57> 274 5 enterocola, sp.n., Biochemical reaction of 279 ” ” ” =p > » from a case of diarrhoea.. 277 +3 insolita, Biochemical reaction @}8) Soonocnobeicnooccnapocooece: Ly ARYL » krusei, Biochemical reaction of 279 53 metatropicalis, Biochemical TEACHONIOL Wee wedscnescsnes 57> 274 » mivea, Biochemical reaction Ob ives ayeingncdiaccs arenes 57, 274 »» parakrusei, Biochemical re- ACHONUOL cee sehen acepneanoseaoe 279 3 paratropicalis, Biochemical TEACLIONJOL 1) 2. cactevinesceeks 57, 274 5 pseudolondinensis, Biochemical FEACHIONYOl..en, acsacssesce aces 277 8 pseudolondinoides, Biochemical MEaCHONsObey reesans ie yaen te 2777 »» pulmonalis, Biochemical re- ACHONNOL gi seagiesset cise acl 57, 274 55 tropicalis, Biochemical _ re- ACEION OL even on ertee scare 57> 274 Monobelea litoraurea, sp.n., female...... 344 Monopylidium gallinarum, sp.n., in Indyantpoultry, . ss... seas sseessn2veee= 164 Montgomeryi group of Cylicostomum ... 401 Mosquitoes, Apparatus for the in- dividual breeding of ... 473 A in the Isle of Man......... 73 ms and other blood-sucking arthropods of the Upper Shiri River, Nyasaland ee toss sen 457 PAGE Mosquito ova, Certain uniform charac- LETS! Oleven ston cacenanepeens sets eee eee 418 Multiple aneurysms in a child ......... 245 Musca domestica as a ‘bush-fly’ in Australia’ 43 secs sc. sisacepe soe eee 93 Newstead, R., and Davey, J. B. Mosquitoes and other blood-sucking arthropods of the Upper Shiri River, Nyasaland .......0.s0 africanus from ” ” Nyasaland 460 x papatasii from India ...... 104 5 sergenti from India ...... 105 Piroplasmosis research by the Liverpool School... scavessccesnedsscceeteneeeetieeten 16 Pistia stratiotes, source of mosquitoes and midges. <:...s.ssssscssuesacconeretaes 314 PAGE Plasmodium falciparum, Indigenous infection Withivigacssss 59 sy is infection, incubation period, 62, 63, 68 “A x infection, incubation period following inoculation, 64, 65 = BS infection, incubation period following bite ieetss. 66, 67 Poultry, Indian, Cestodes in ............ 161 Prevalence and character of tubercu- losis in Hongkong............ 213, 227, 381 Probezzia pistiae, sp.n., adult ......... 364 ra Bs Ree ATV sc isevesess's 368 “A i oo PUpabsccsccees 367 rr stephensi, sp.n., male ...... 369 Protozoa, Symptomless carriers of...... 117 Protozoological research by the Liver- IGEU CUO Ses sc. ssc edecspseucassoecsssot 45 Pseudolondinensis group of Monilia, Biochemical reactions of................ 277 Psorophora posticata ..s.sccsveseceeeeess 446 Psychodidae from Nyasaland............ 460 Pupipara from Nyasaland ............... 461 Queensland, Arneth count in hook- worm-infected children in............. 49 Radiatum-elongatum group of Cylicos- MUI Mertesct Tabanidae from Nyasaland ............ 460 TGBAnUS GPPICOTUS enneacesententtrecseeenes 460 9, taentola. Var. VATIAtUS.....0.000 460 LiGenta MAPN ” cece vocteechvc soeeeanecteak 255 5. mE DULCALG (os cracas vias cesae ocdes eee 255 % eecbraes Collinte-s.cecitrsen tions 256 a gp) WAGER as chtae scan sceaees 255 Taentorhynchus titillans.........cceeeceeees 445 Tartar emetic in treatment of Oriental 70) Hee ERROR OT Se ee 107 Tetracanthum-coronatum group of Cyli- COSCOMUTE paca ncons ca de- Soeh aoe: Tee ena 397 Theobaldia annulata in the Isle of IVEaD sconce cnsctecseeststaseer ee rasealeees 75, 88 Thysanosoma giardi in a new host....... 405 Tonsillar nocardiomycosis, Case of, in West Atricd a cen. ti. arcane 282 Trichiuris in symptomless carriers 123 Trichonema genus, question of nomen- clatnre'(fcotnbte)).....0s.-essctaces ace Trigonotylus brevipes Tropicalis group of Monilia, Bio- Trypanocidal effect of bismuth ...... 433 * effect of phenylglycine amido arsenate of sodium on T. brucet in rats and I. rhodesiense in mice Trypanosome brucei in rats treated with phenylglycine amido arsenate of sodium ... 427 33 rhodesiense, Case infected with, treated with * Bayer 205 © ceseenamene 479 s rhodesiense in mice treated treated with phenyl- glycine amido arsenate 427 of sodium) 2ycc.-s5-eseue 427 ‘Trypanosomiasis research by the Liver- pool Schools... .<.cceussdes-seeeeeeee 16 Tsetse-flies, new, from the Belgian. Congo .;...05c04.--09s sa0ss ene 95 Tuberculosis complicated by broncho- moniliasis in the Gold Coast 5. Seesceseueeres 53, 285 35 Education as a weapon against........ Fepeeae as 224 in Hongkong ...213, 227, 381 = Morbid anatomy in cases Of ccondeceenandeneeeanatee 381 es Portals of entry and mode of spread of, 214, 219, 227 = Possible congenital infec- tion With, \....<-:-facssus Rai Racial susceptibility to... 218 Ulecrative granuloma, Notes on ...... 413_ Unusual breeding-places of Stegomyia fasciata in Australia.,.<.--s-sescssueuns gI Venezuela, Notes on Culicidae col- lected iN... o..:cc-ssayasssaeede eee - 445 Venezuelan oilfield, Malaria on a ...... 435 Water-bugs destructive to S. calopus TAF VAC oc 11. n0ee cee coneseeeeeeee eee eee 303 West Africa, Fungal infections in....... 271 West African Ceratopogoninae ...177, 313 NXenopsylla cheopis on rats at Liverpool 290 »» breeding-places of... 295 Yellow Fever research by the med. School, |. ssseuscentermeee gent > saan 28 Yorke, W. Treatment of a case Rhodesian sleeping sickness by the preparation known as ‘ Bayer 205” 479 Yorke, W., and Adler, §. Note on a case of leprosy Joes Sys Seee see eee 269 Yorke, W., and Southwell, T. Lap- peted Anoplocephala in horses......... 249 Young, C. J. Natural enemies of Stegomyta calopus ......2000cs0eeeeeeeeres 301 Young, C. J., and Gordon, R. M. The feeding habits of Stegomyia RBM Seco sara sacc isso rss vasiedss 265 PAGE Zaitha spp: destructive to S. calopus larvae Zebra, Anoplocephala rhodesiensis in.... 249 Zschokkeella genus, Synonymy of ...... 455 =n gutneensis from an African Eat) 2s; scenes 167 % 55 Synonymy of.... 455 _INDEX OF GENERA, SPECIES AND VARIETIES NEW TO SCIENCE PAGE PI RUIATIA BUSITAIIS .......0000eeceeceseeeee 407 Atrichopogon africanum .....0...ccseveee 334 Atrichopogon chrysosphaeratum ......... 337 Atrichopogon elektrophacum............++ 335 Atrichopogon homoium .........-0000000000+ 338 Atrichopogon perfuscum .....s00.0eceeeees 337 BRENT E cove ccaie ses eec cafe seses sass xes< 361 IMPEDES) COTSONL.«....ce--2s0scesecesnerses 324. Culicoides inornatipennis, var. rutilus... 326 CUIECONAES THZETIGE ......0sccc.0eceoeveeees 325 Cylicostomum labiatum, var. digitatum 401 Dasyhelea boothi .........ccevescceccseseeee 331 OG ETT CG LS EE 196 Dasyhelea flaviformis ..........c000eeeeees 201 TREBLE Z USCA cn .0 sce cocaseesencccceenseste0 204. Dasyhelea fusciformis ...cccccccceseceeeees 209 " Dasyhelea fusctpleurts ...........c.000e0e 200 Dasyhelea fusciscutellata .........0.00000+ 187 Dasyhelea inconspicuosa .......0s00cece00s 191 Dasyhelea luteoscutellata .........0002.0+06 19I WTARVDCIEA BI QETIAL ....c00..eccecaseeescees 329 Dasyhelea nigricans ......0....cccocvesseees 194 PAGE Dasyhelea migrofUuscd ..secrervseereerevees 207 Dasyhelea pallidihalter ......ccccceeeeees 184 Dasyheled retorta ..10..ccsnecenveceensseores 333 Diasy Bele atts canst casccanctagescea nnn’ 189 Dicrobezzia migritibialts ......2s0eeeesees 371 DUEDESREMIEPU | scanvecedesater se se--eeraa= sr 169 Eukraiohelea genus ........00cssseeveeeees 347 Eukraiohelea versicolor ..ssccccseveeseves 351 Glossina fusca, var. congolensis.........++. 99 GIOSSIMAISCHOEAN .. 2ses0p>--0-20-ccnsesceee 95 RET PIAIOCREOSOM A sosotaccanssaees>aon) “< { Yd + mn, \ f at wy thks a “ ad tye ie ie vias. ay ” ize Sierra Saty SAY rus ve Sy 2 ‘ Fey » bebe Ve 2b aL nde twin 4 i w x > = % att 1 gt > ay 9 as ’ a, 5649 = = es - . . . * + » yr cet te has hig a ae diva as1759d Ana, i- Tur Late Dr. A. J. CHALMERS LIVERPOOL SCHOOL OF TROPICAL MEDICINE SCIENTIFIC RECORD COMPILED BY DOROTHY ALLMAND Librarian of the Liverpool School of Tropical Medicine PLATES J—IX The new laboratories of the Liverpool School of Tropical Medicine were formally opened on 24th July, 1920, by Lord Leverhulme, Honorary Vice-President and former Chairman of the School. The building had been placed, in 1915, at the disposal of the Military authorities for the purposes of a hospital, and was used by them until 1919, for the treatment of patients suffering from tropical diseases, mainly malaria and dysentery. The hospital, . which consisted of about 200 beds, was under the charge of Lieut.-Col. J. W. W. Stephens, R.A.M.C., Professor of Tropical Medicine, and during its four years’ existence more than 3,000 cases were admitted. Subsequent to evacuation by the Military authorities, the decoration and equipment of the laboratories were put in hand, and to-day the School affords excellent conditions and facilities for instruction and research. The laboratories are situated in the University grounds, and close to the Royal Infirmary, in which the School has its Tropical Ward and adjoining clinical laboratory. In addition to the basement, which contains the Photographic Department and large storage rooms, there are four floors. The ground floor comprises the Lecture Theatre, with accommodation for about seventy students; the Library, where, besides text-books and miscellaneous scientific publications, over one hundred current medical journals from all parts of the world can be consulted; and the Museum, a spacious room, 80 feet by 69 feet, with preparation room adjoining. 2 On the first floor are the Departments of Tropical Medicine and Entomology, the latter with its library of specialised literature, and six research rooms. The second floor comprises the excellently lighted class laboratory, 69 feet by 58 feet, and four research rooms devoted to the Department of Parasitology, while the third floor has a large research laboratory and two smaller research rooms. On the roof are an Insectarium, a mosquito-proofed house, and the animal houses. The cccasion of the official opening was marked by the issue of a volume* which traces the history of the School from its foundation down to the year 1920. The main purpose of this publication was to perpetuate the names of those who have been closely associated with the School in its varying activities in the past, and this aspect may here be briefly summarised before proceeding to an account of its scientific activities which, in the ‘Historical Record,’ are merely outlined. The Liverpool School of Tropical Medicine was founded in 1898 by the late Sir Alfred Lewis Jones, K.C.M.G., a prominent Liverpool ship-owner, who, fired by an appeal for the study . of tropical diseases issued by the late Rt. Hon. Joseph Chamberlain, then Secretary of State for the Colonies, offered to the President of the Royal Southern Hospital the sum of 4350 for three years, to promote the special study of such diseases. Quarters were obtained in the Pathological Department at the University, and a Dean was appointed in the person of Professor Rubert Boyce, to whose tireless energy much of the early success of the School was due. In February, 1899, Dr. H. E. Annett was appointed Demonstrator in Tropical Pathology ; in April, Major Ronald Ross accepted the post of Lecturer, and the School was officially opened; and in May teaching commenced. : The early history of the Liverpool School is a paies of struggle; it was not founded by the Government, as was the London School; it had no grant, no assured income, nor even, at first, Government recognition. The School early turned its attention to research, and in the summer of 1899 the first of its expeditions was despatched to Sierra Leone. During the next fourteen years funds were * Liverpool School of Tropical Medicine: Historical Record, 1898-1920. Ar the University Press, 1920. viii, 103 pp., 36 plates. 10/6 post free. 3 collected to equip and maintain more than thirty expeditions to various tropical regions, including West and Central Africa, Brazil, and the West Indies. In the meanwhile, at home the School was enlarging its scope and consolidating its position. In 1900 the Government gave it full recognition by placing it on the same terms as the London School with regard to newly-appointed medical officers and their courses of training, and four years later further recognition was received in the form of a financial grant from the Colonial Office, which grant was later increased and has continued to the present day. In Igor a Lectureship and Fellowship were founded, in commemoration of the work of the late Walter Myers, who died of yellow fever in Brazil while serving with the Fourth Expedition. Major Ross was appointed to the former, and the latter was filled by Dr. J. E. Dutton, who, four years later, died in the Congo while investigating tick fever. In 1903 the Alfred Jones Chair of Tropical Medicine was founded, the first holder being Major Ross, while Dr. J. W. W. Stephens, who had joined the staff in the previous year, was appointed to the vacant Walter Myers Lectureship. In 1905 another important Lectureship was founded, namely, that of Economic Entomology, which was accepted by Mr. R. Newstead, of Chester. As the work and influence of the School increased, many new appointments were made, so that now, at the age of twenty-two, the School has three University Chairs,* namely, Tropical Medicine, Entomology and Parasitology, filled by Professors Stephens, Newstead and Yorke, respectively, assisted by a staff of eight Lecturers. In 1903 the School moved into new laboratories, and in the following year a Diploma in Tropical Medicine was instituted. In 1904 the Runcorn Research Laboratories were established for the purpose of conducting investigations on the large collection of trypanosomes and other protozoa, which had been amassed by members of the expeditions on various occasions. These labora- tories were first placed in the charge of Dr. Wolferstan Thomas, and during their ten years’ existence much valuable research was done there. * A fourth Chair, that of Tropical Diseases of Africa, has now (1921) been established, and is held by Professor B. Blacklock. : 4 In 1906 was issued the first number of the Axnals of Tropical Medicine and Parasitology. This publication had been preceded by a series of twenty-one Memoirs, mostly reports of the School’s expeditions, In 1911 the Yellow Fever Bureau came into being with Dr. Harald Seidelin as Director. As the name implies, the Bureau was established for the purpose of promoting the study of the many problems surrounding this disease. A journal was issued, the Yellow Fever Bulletin, of which three volumes were published. Clinical instruction to students of the School had formerly been ~ given in a special ward at the Royal Southern Hospital. This hospital being at some distance from the laboratory, arrangements were subsequently made with the Royal Infirmary for the erection of a new tropical ward in the Infirmary grounds. The ward, which contains ten beds, with an adjoining clinical laboratory, was opened in 1914. On the arrival of the Fifteenth (Yellow Fever) Expedition at Mandos in 1905, it was found necessary for the work of the Expedition to establish a laboratory of a more or less permanent character. The Mandos Research Laboratory remained in existence until January, 1909, when, owing to the return of its Director, Dr. Wolferstan Thomas, to Liverpool, it was closed. In June, 1910, Dr. Thomas returned to Manaos and opened the present laboratory. In 1914 it was decided to extend the activities of this branch of the School; the outbreak of war, however, caused all developments to be deferred until 1919, when three research assistants were appointed to the laboratory. With a view to carrying on research work in tropical medicine, the School desired to establish a permanent laboratory on the West Coast of Africa. Funds were available, through the munificence of the late Sir Alfred Jones, and a suitable site on Tower Hill, Freetown, Sierra Leone, having been placed at the disposal of the School by the Colonial and War Offices, the laboratory is now in course of erection. dnnals. Trop. Med. & Parasitol., Vol. XV 5 PLATE I FIRST LABORATORY OF THE SCHOOL 5 The following brief record of the research work of the School is dealt with under subjects; no attempt at a critical estimate has been made. MALARIA AND SANITATION The first Expedition left England in July, 1899, for Sierra Leone, and consisted of Major Ross, Dr. H. E. Annett, Dr. R. Fielding-Ould, Mr. E. E. Austen, officially appointed by the British Museum, and Dr. Van Neck, delegate of the Belgian Government to the School. The purpose of the Expedition was to study malaria in man, and a report of the work accomplished was published in 1902 as Memozr II. The investigators discovered two species of Anopheles in Freetown, namely, A. /unestus, Giles, and A. costalis, Loew, and by dissection established the presence of blasts (sporozoites) in them. They concluded that both A. costalis and A. funestus are hospitable to the human Haemamoebidae; that A. costalis is hospitable to all three. of the human species, and A. funestus certainly to H. malariae, and probably H. ‘vivax; no observations were made regarding the connection of A. /umestus with H. praecox. They further pointed out the difference between Culex and Anopheles, and studied more especially the bionomics of the latter. ‘Precautions against the bites of gnats’ and ‘ operations for reducing the number of Axzopheles’ were fully considered. Dr. Fielding-Ould continued the work of the Expedition at Freetown after the other members had returned to England, and he also prosecuted his researches at Accra and Lagos, his report of which is included in Memoir Il. At Freetown six of twenty-nine Anopheles dissected showed parasites. Seventeen Culex gave negative results. At Accra fifty-two Culex were negative. At Lagos thirty-seven Anopheles were dissected and zygotes found in one. The reports dealt mostly with the sanitary survey of the districts. * In 1900 a third Malaria Expedition was sent to Northern and Southern Nigeria. This Expedition, which was composed of Drs. H. E. Annett, J. E. Dutton and J. H. Elliott, spent six months in Nigeria, and in Memoirs III and IV reports were published of the work done, the former relating to malaria and the latter to filariasis (see p. 37/). Seven of two hundred and eighty-one Anopheles 6 dissected at Bonny were found to be infected with malaria; it was noted that it was difficult to decide what was the exact type of parasite present. The observation made by Koch and by the Royal Society’s Commission that native children are infected with malarial parasites to a large degree was confirmed ; the incidence of infection is given in the following table. It was also proved that Tapre showing the total number of children examined and found infected throughout Nigeria. Ages Number examined Number infected Percentage infected o-5 220 114 518 5-10 } 108 27 25°0 1o+ 40 4 10°O quartan and simple tertian parasites exist in West Africa, a fact contrary to the experience of the Royal Society’s Commission of the previous year. The bionomics of Anopheles were studied still further and a series of experiments in propagation carried out, confirming and elaborating the discovery of the previous Expedition that the female mosquito requires a meal of blood both for fertilisa- tion and for the development of the ova. In their recommendations concerning prophylaxis they advocated (1) the segregation of Europeans at a distance of about half a mile (a principle already put forward by the Royal Society’s Commission on Malaria also at work in West Africa), and (2) the surface drainage of areas around their quarters. In an appendix they gave charts and descriptions of cases of hyperpyrexial fever, first described by Thompstone and Bennett. The hyperpyrexial stage lasts one to three weeks, and is followed by very extended lysis. An exhaustive account was given in the second report (AZemoir IV) of the mouth parts of the female A. costalis, and, in an appendix by Theobald twenty-five mosquitoes were described of which nine were new species, namely, Stegomyia ivritans, S. nigricephala, Culex duttoni, C. decens, C. pruina, C. invenustus, C. nebulosus, C. vima and C. invidiosus (with figures). In June, 1901, Major Ross and Dr. Logan Taylor went to Freetown to organise an anti-mosquito campaign. Work was 7 commenced by engaging the services of between thirty and forty men, who were divided into two gangs: the Culex gang and the Anopheles gang. ‘The former collected from private houses all the broken bottles and buckets, empty tins, etc., in which S/egomyia and Culex breed ; the latter drained the pools and puddles in which Anopheles breed in the streets and backyards of the houses. Reports on the progress of this campaign were published as Memoir V, Parts 1 and 2, and later Ross issued a small book: ‘Mosquito Brigades, and how to organise them.’ In September, 1901, Dr. Dutton went to the Gambia and inspected the conditions of health there, with the result that the Colony organised measures similar to those in operation at Freetown (Memoir X). About 32 per cent. of the infections examined Tasxe showing the total number of children examined and found infected throughout the Gambia. Ages Number examined Number infected Percentage infected o-5 78 64 820 5-10 22 20 gt’o 1o+ 13 7 53°83 showed quartan parasites, and about a third of the malignant tertian cases showed crescents. The simple tertian parasite was found three times only. Dutton noted the universal infection of canaries with Haltéeridium, which was also found in other birds. Of twenty-four A. funestus dissected, one contained sporozoites and one zygotes. A. costalis was found breeding in boats, street drains, wells, tubs and barrels, and in tidal water containing 1°7 per cent., salt, together with C. thalassius. It would appear that A. fumestus and its varieties are rural mosquitoes, while A. coséalis is essentially town bred and capable of utilising any small collection of water for breeding purposes. In an appendix, descriptions of eighteen species were given by Theobald, including A. costalis, var. melas; A. funestus, var. 8 umbrosus; A. funestus, var. subumbrosus; Stegomyia albocephala, sp.n.; Culex annulioris, var. gambiensis, v.n.; C. anarmostus, sp.n.; C. thalassius, sp.n.; C. euclastus, sp.n.; Lasioconops poicilipes, gen.n., sp.n.; Corethra ceratopogones, sp.n. Eight specimens of G. longipalpis var. tachinoides were taken. Dutton recorded the finding of a filarial embryo in the thoracic muscle of C. anarmostus. It was during his researches into the blood parasites of the Gambia that Dr. Dutton made a discovery of the highest scientific importance, namely, the identification for the first time of a trypano- some in the blood of a man, a patient of Dr. Forde. This parasite was subsequently (see p. 16) shown to be the cause of sleeping sickness, and was named Trypanosoma gambiense, Dutton, 1902. In 1901, Dr. C. Balfour Stewart was sent to the Gold Coast, to conduct operations similar to those carried on at Sierra Leone, while early in 1902 Major Ross again visited Freetown to ascertain by a thorough inspection the results of the previous Expedition. In the autumn of this year, Dr. Logan Taylor proceeded to Cape Coast Castle, Gold Coast, and reported upon the sanitary conditions prevailing ithere, with suggestions as to their improvement. (Memoir VIII). In September, 1902, by request of the Suez Canal Company, Major Ross went to Ismailia to investigate the causes of the prevalence there of malaria, and to recommend measures for its prevention. Axopheles and Culex were found breeding in water containing o'g per cent. of salt. He concluded that the majority of Anopheles which caused malaria in Ismailia came from the marshes in immediate proximity to the town. Prophylactic measures based on his recommendations were commenced immediately (AZemozr IX). In February, 1904, sixteen months later, Professor Boyce si: Ismailia; from statistics furnished in his report (Memoir XII), appeared that the number of cases of malaria had fallen from 1 ‘si in 1902, to 209 in 1903. During 1904, two Sanitary Expeditions were despatched to West Africa: one to Bathurst, Conakry and Freetown, and the other to the Gold Coast. Reports were published in 1905 as Memoirs XIV and XV. In May, 1906, Professor Ross went to Greece in order to advise Annals Trap. Med. & Parasitol., Vol. XV PEA Ronen Ase i” ss LM ib pas cer af Be x Sa oe __ al SECOND LABORATORY OF THE SCHOOL 9 the Lake Copais Co. with regard to anti-malaria measures. In the following year, at the request of the Colonial Office, he visited Mauritius for the same purpose; his report, entitled ‘The Prevention * of Malaria in Mauritius,’ was published in 1908 (Waterlow & Sons). In 1908, the Twenty-first Expedition of the School, consisting of Professor Newstead, Dr. W. T. Prout and Dr. Alan Hanley, was despatched to Jamaica. Reports were published in the Annals of Tropical Medicine and Parasitology, Vol. Ill, pp. 421 and 471, the first, dealing with medical and economic entomology, by Professor Newstead (see p. 39), the second, on malaria, by Dr. Prout. From data furnished by Dr. Neish, of Spanish Town, the frequency of the various species of malaria parasite was found to be approxi- mately: simple tertian 54 per cent., malignant tertian 36 per cent., quartan about 1 per cent., and mixed infections about 8 per cent. The occurrence of several cases of blackwater fever was noted. The enlarged spleen rate in different parishes varied from 0 to 60 per cent. Prout found that the average percentage of malarial deaths to total deaths was nearly 20 per cent., that in four years the total admissions to hospitals from malaria had increased by 55 per cent., and that over 33 per cent. of the total admissions were due to malaria; it was reported that the annual cost of treating malarial patients was over 46,000. Various anti-malarial measures were recommended. In 1909, Boyce visited Jamaica, and reported upon the work done by the Commission appointed by the Governor in October, 1909, to deal with the malaria problem (Azmzals, Vol. IV, p. 233). It was recorded that one-half of the island, namely, those parts above 1,000 feet, was practically free from malaria-carrying mosquitoes. Ross and D. Thomson, working in Liverpool, published a paper entitled ‘Some Enumerative Studies on Malarial Fever’ (Proc. Roy. Soc., B., Vol. LXXXIII, p. 159, and Aznals, Vol. IV, p. 267). Some of the conclusions were: (1) No fever exists unless the parasites exceed from 500 to 1,500 per c.mm.; (2) the parasites tend to remain continuously in the blood in small numbers between the febrile relapses; (3) close correlation between the number of parasites and the amount of fever caused by them; (4) studies on quinine gave a numerical estimate of its effect, a few days’ use of the drug reducing the parasites by from 50 per cent. to 80 per cent. ; 10 (5) crescents apparently require eight to ten days for development; quinine affects their numbers only by destroying the generating cells. Thomson, who subsequently investigated the life history of crescents (Avnals, Vol. V, p. 57), concluded that these do not live ° for more than a few days in the peripheral blood. They are replenished from surviving asexual forms, and quinine has no action on crescents but only on the asexual source of supply. He concluded (Azmals, Vol. VI. p. 223) that administration of quinine in doses of 20 grains daily for three weeks is almost certain to destroy both the asexual and sexual parasites. Ina study of the leucocytes in malarial fever (2éd., p. 83), he stated that malaria could be diagnosed by the leucocytic formula. In 1912, J. G. Thomson and McLellan confirmed Bass’s observa- tions on the cultivation of malarial parasites. In twenty-four hours P. falciparum was found to undergo segmentation, the maximum number of merozoites counted being thirty (Azzals, Vol. VI, p. 449). In the case of P. vivax (Annals, Vol. VII, p. 153), sixteen merozoites were produced. These cultures of P. vzvax differed from those of P. falciparum in that there was no tendency to clumping. Further experiments in the cultivation of P. falciparum and P. vivax were made (2dzd., p. 509), when it was noted that the optimum temperature for cultures was 38° C. Sinton, investigating Uriola’s test of malarial pigment in the urine (Annals, Vol. VI, p. 376), concluded that it is almost” impossible to exclude extraneous pigment. In 1912, Dr. David Thomson was sent to Panama to study certain malarial problems with Dr. James, Chief Assistant Physician to the Ancon Hospital. The first part of his report (Annals, Vol. VII, p. 125) dealt with the sanitation in the Canal Zone, Trinidad and British Guiana; the second (Azmals, Vol. VIII, p. 85) with the origin and development of gametes in malignant tertian malaria. He noted that they develop chiefly in the bone marrow and in the spleen, the period of incubation being about ten days. While examining a malarial blood film sent from the Central Provinces of India, Stephens was struck by the peculiar appearance of the parasite (Proc. Roy. Soc., B. Vol. LXXXVII, p. 375, and Annals, Vol. VIII, p. 119). It exhibited the following peculiarities : II (1) it was extremely amoeboid ; (2) the cytoplasm was scanty ; (3) the nuclear protoplasm was out of proportion to the volume of the parasite. It differed from P. falciparum by its amoeboid activity, and from P. vivax by its smaller size, the delicate nature of its amoeboid processes; the irregularity of its chromatin and the rarity of typical ring forms. Stephens proposed to name this parasite P. tenue. Later he described peculiar forms of a malaria parasite in a blood slide from the Gold Coast (Azmals, Vol. IX, p. 169). In addition to these, large and apparently quite normal quartan parasites occurred, and it was possible to trace a transmission from normal ring forms to those in which chromatin particles or strands without any protoplasm were seen in the red cells. In 1917, the School was asked by the War Office to undertake investigations into the treatment of malaria; the results of this work were published in thirty papers (Aznals, Vols. XI-XIII). No case was considered to be malaria unless parasites were found. The results of treatment were in all cases controlled by daily microscopical examinations combined with the clinical record. Simple Tertian. The investigators established: (1) that intra- muscular injection of quinine bihydrochloride was an effective method of treating a malarial attack, a matter which had been one of considerable dispute in the medical press just previous to this work. (2) That for the palliative treatment of malaria, that is, for keeping a person free from relapses over long periods, it was better to give a certain total amount of quinine on each of two consecutive days than on each of six days in the week: thus, 60 grains administered as 30 grains for two days gives a better result than the same amount administered as 10 grains for six days. (3) The best palliative result was obtained by administration of 45 grains on each of two consecutive days weekly over a period of two months. (4) They found that a certain treatment may give a certain ‘curative’* result on one occasion, while the same treatment repeated on another occasion might give a quite different result. (5) Novarsenobillon was found to be as efficacious as quinine in the treatment of paroxysms, but its curative effect, like that of quinine, was practically nil. (6) The best ‘curative’ result was obtained by * The term cure was used to signify no relapse within an observation period of 60 days after cessation of treatment. Iz the administration of Lzguor arsenicalis, minims 30, daily over a period of eight weeks in combination with two initial intramuscular injections of quinine bihydrochloride, grains 15, on two days only. Owing to the Armistice, these observations were not repeated. The intravenous injection of quinine was found to have no real curative effect. The relapse period after treatment of eight hundred simple tertian cases was recorded (Azmals, Vol. XIII, p. 125). It is essential to recognise that these figures were based on an observation period of sixty days. The incidence is shown in the accompanying graphs. It will be seen that of those cases that relapse the majority do so in the first twenty days after cessation of treatment. Further, the time of occurrence of the paroxysms was noted in one thousand cases (Aznals, Vol. XIV, p. 365). The majority occurred at 2 p.m. with the conditions of life under which the patients (soldiers) were living; over go per cent. occurred during the hours of activity, that is, between the hours of 7 a.m. and 6.59 p.m. Malignant Tertian. Neither a single nor a series of six intravenous injections of quinine bihydrochloride (grains 10 to 15) caused the disappearance of parasites, either trophozoites or gametes, from the peripheral blood, whereas in the case of simple tertian the disappearance was rapid. Under quinine treatment, grains 30 to 45 daily, crescents did not persist in the peripheral blood in the majority of cases for more than three weeks. How long they persist without quinine was not determined. {n July, 1919, Blacklock and Carter recorded the experimental infection, for the first time, of Anopheles plumbeus with P. vivax. Experiments were made (Azmals, Vol. XIII, pp. 187 and 413), with the result that the observers were able to obtain infections of the gut and salivary glands of laboratory-bred A. plumbeus at a temperature of 28° C.; at room temperature gut infection only was obtained. Infection of the gut was also produced with P. vivax in the case of A. difurcatus at 28°C. Later experiments (Axmals, Vol. XIV, p. 275) with A. flumbeus resulted in gut infection with oocysts of P.. falciparum at 28° C. : Percentages, GrapH 1. Percentage of total relapses in each 20-day period. 60 8=—680 Days after cessation of treatment. 8) GrapPH 2. Percentage of cases treated which relapse in each 20-day period. GraPH 3 Percentage of cases treated not having previously relapsed which do so in each 20-day period. 14 BLACKWATER FEVER In 1907, the Nineteenth Expedition, consisting of Dr. J. O. Wakelin Barratt and Dr. Warrington Yorke, was despatched to Nyasaland to study blackwater fever, a report being subsequently published in Anxals, Vol. III, p. 1. The object of the investigators was to trace out some of the internal processes, the terminal event of which is the appearance of blackwater, believing that in that way many obscure points in connection with the causation and treatment of the condition would be cleared up. The first point was to determine the action of quinine, acid and alkali upon the red cells during blackwater fever. They found that haemolysins, present in the blood, played no part in the production of black- water. It was considered that the suppression of urine is due to a mechanical blocking of the renal tubes by the formation of large, firm, coarsely granular casts in the ducts of Bellini. In a later study (Aznals, Vol. V, p. 287), on the suppression of urine in black- water fever, Yorke and Nauss re-investigated the mechanical theory and found that it is considerably facilitated by any factor which tends to lower the blood-pressure, and by that means the secretion of water by the glomeruli, but that if the blood-pressure is kept up by the injection of saline solutions, the tendency to suppression is decreased. Arising out of these latter experiments, the passage of haemoglobin through the kidneys was studied by Yorke (zdid., p. 401), who was led to consider that haemoglobin is excreted by the renal epithelium rather than filtered through the glomeruli, and that the amount of haemoglobin eliminated into the urine is dependent upon the activity of the epithelium lining the renal tubules. It was shown by Simpson (Azmals, Vol. VI, p. 313) that the haemoglobin liberated from the red cells in malaria escapes in larger quantities by the faeces than by the urine. The study of haemo- globin metabolism in blackwater fever was continued, and a report made (io-Chemical Journal, Vol. V, p. 378) on the quantitative estimation of urobilin in the excreta. In later observations on haemolysis in malaria (Azmals, Vol. VI, p. 231), Simpson concluded that the serum of malarial patients may possess the power of haemolysing normal red blood cells. The haemolytic effect could Annals Trop. Med. & Parasitol., Vol. XV PLATE Ill FRONTAGE OF THIRD LABORATORY OF SCHOOL 15 not be obtained at all periods of the paroxysm, nor in every case; it appeared to be produced at the period of sporulation, and rapidly disappeared. Simpson and Edie (ébid., p. 443), observing the excretion of urobilin in animals and man, found that an increase may occur after the administration of quinine in doses of 10 to 30 grains a day, and that a similar result follows injection of blood pigment or haemolytic drugs. Experiments were devised by Barratt and Yorke (Azzals, Vol. VIII, p. 509) for examining the relation of bile pigments to haemoglobin. Experimenting with rabbits, they found that consequent upon intravenous injection of haemoglobin solution there was a distinct and immediate increase not only in the concentration of the-bile pigment, but also in the amount of bile pigment excreted. Two hypotheses were advanced to explain this increase: (1) that the haemoglobin injected is actually converted by the liver into bile pigment; or (2) that it merely stimulates the liver cells to an increased production of bile pigment. Stephens (Thompson-Yates Lab. Reports, Vol. V, Pt. 1, p. 193) recorded that blackwater fever occurred in eleven of twenty-two of the United States. An account was given of the distribution of the disease, and an extensive bibliography appended. An analysis of ninety-five cases showed that when the blood was examined before the onset of blackwater, malarial parasites were present in 95°6 per cent. of cases, whereas on the following day the remarkable fall to 17°I per cent. was the result. On the day of blackwater itself the figure was 61'9 per cent. A series of studies in blackwater fever were made by Stephens (Annals, Vol. VII, p. 479). The subject was considered under the following headings: (1) malarial parasites; (2) pigmented leuco- cytes; (3) post-mortem examinations; (4) influence of malaria; (5) relationship to species of malaria parasites ; (6) effect of period of residence; (7) seasonal prevalence; (8) correlation between malaria and blackwater statistics; (9) second attacks. A schedule for recording cases of blackwater was devised and recommended (Annals, Vol. VIII, p. 639). The results were recorded (Azzals, Vol. IX, p. 201) of a statistical examination of the respective times at which quinine was given and blackwater occurred. Graphs were published showing that correlation existed between the time of 16 taking quinine and the onset of symptoms, but this did not necessarily imply any relationship of cause and effect. Data on the duration of haemoglobinuria were collected (Axmals, Vol. IX, p. 539). In one hundred and sixty-seven records it was found that the duration was not more than twelve hours in a quarter of the cases, not more than one day in half the cases, and not more than two days in three-quarters of the cases. Finally, the importance of furnishing population statistics in connexion with cases of black- water fever was emphasised (Azmals, Vol. X, p. 345). PIROPLASMOSIS At Runcorn, Breinl and Hindle studied the morphology of*Piro- plasma canis (Annals, Vol. II, p. 233), and worked out the nuclear details of the parasite. Later, Breinl and Annett (2é7d., p. 383) concluded that the haemolysis in Pivoplasma canis is not due to a formation of a specific haemolysisin or isolysin, but to the mechanical disintegration of the red blood corpuscles after the escape of the parasites from them. Barratt and Yorke found that in piro- plasmosis the haemoglobinuria was attended with and dependent upon haemoglobinaemia. Of one hundred and forty-three cattle examined at Sierra Leone (Azzals, Vol. IX, p. 418) about 5 per cent. were found to be infected with P. bigemznum, while Theileria mutans was encountered in between 20 and 30 per cent. TRYPANOSOMIASIS In view of the great importance of the discovery of the trypano- some in man by Dr. Dutton* in 1901, described and named by him T.. gambiense (Thompson-Yates Reports, Vol. IV, p. 455), an Expedition was sent out to the Gambia and French Senegal in September, 1902, in charge of Dr. Dutton and Dr. J. L. Todd, for the study of Trypanosomiasis. While these investigators were at work abroad, the first case, that of a European who had returned with Dr. Dutton in 1901, remained in Liverpool, and Dr. Annett infected monkeys and 25 per cent. of tame rats successfully, but did not succeed in infecting tame mice, rabbits or guinea-pigs. One of the infected monkeys died, but the other recovered, and no parasites * For the history of the discovery see Boyce, Ross and Sherrington, Lancet, Feb. 21, 1903. a7 could be found by sub-inoculation into rats. The results of his work were incorporated in the report of the Expedition (Memoir XI). On reaching the Gambia, one thousand and forty-three natives were examined, the majority of whom were children or young adults, and apparently healthy; six were found to be infected. Trypanosomes were also found in the blood of a quadroon. Clinical descriptions were furnished in the report of these first eight cases of trypano- somiasis, of which those of the European and the quadroon terminated fatally after a duration of about eighteen months. Of thirty-six horses examined, ten were found to be infected with trypanosomes. Transmission experiments were made with Glossina palpalis and later with Stomoxys, but with negative results. A series of inoculations were made of the human and equine trypano- somes in experimental animals, the results of which led Dutton and Todd to the conclusion that the parasites were not of the same species; the Gambian horse trypanosome was subsequently named T. dimorphon. Several new species of flagellates occurring in birds, mice, tortoises, etc., were described in this report, including T. johnstoni, T. mega and T. karyzeukton. Prior to the return of this Expedition, the discovery of trypano- somes in the cerebro-spinal fluid of cases of sleeping sickness in Uganda by members of the Sleeping Sickness Commission of the Royal Society, caused the subject of trypanosomiasis to assume great importance. At the invitation of King Leopold, an Expedition was sent in 1903 to study sleeping sickness in the Congo Free State, consisting of Drs. Dutton, Todd and Christy. The results of these investigations were incorporated in Memoir XIII, and illustrated the occurrence and distribution of trypanosomiasis, described the symptoms of the disease in all its stages, both in Europeans and natives, and showed how sleeping sickness, so-called, is related to trypanosomiasis as a symptom of that disease. They first stated that they were unable to find any difference between the trypanosome occurring in cases of sleeping sickness in the Congo and 7. gambiense. There was a very evident clinical connection between cases with-only very slight symptoms (trypano- soma fever) and advanced cases of ‘sleeping sickness.’ In twenty- five of thirty-eight cases they found parasites in the cerebro-spinal fluid, adopting Quincke’s new method of diagnosis by lumbar 18 puncture. They infected rats, mice, rabbits, guinea-pigs, and studied the morphology of the Congo and Gambian trypanosome in these animals. They noted that about 50 per cent. of such inocula- tions failed, and that they did not succeed in infecting two dog-face monkeys (Cynocephalus species). In the combined areas of Leopoldville, Boma, Matadi and the Cataract Region, among a total of 1,172 persons examined, 8°8 per cent. were infected, while of these latter 55 per cent. had been diagnosed as cases of sleeping sickness. In the Gambia, the previous Expedition had examined 1,043 natives, of which six only harboured trypanosomes, and showed no definite symptoms. Numerous lumbar punctures were made, and it was noted that in many cases the trypanosomes never find their way into the cerebro-spinal fluid, and in those cases in which they do they are more likely to be found towards the termination of the disease; if they gain access early in the disease, mania and other cerebral symptoms are more likely to be prominent, but their. entrance is in no way correlated to the commencement of the fever or other symptoms. In two later papers (Memoir XVI, p. 97, and Memoir XVIII, p. 1) on gland puncture in trypanosomiasis, the observers favourably compared this with other methods of demonstrating the presence of parasites. Following the work of Greig and Gray, they concluded that by gland puncture, cases infected with trypanosomes could be recognised at a much earlier period than hitherto. They also, for the first time, observed a phenomenon frequently seen in cases of trypanosomiasis, namely, auto-agglutination of the red cells. The distribution of sleeping sickness in the Congo was subsequently studied (Memozy XVIII), it being concluded that the increase during recent years was due, in a great measure, to the increase in travel following the opening up of the country. In a subsequent report (Azzals, Vol. I, p. 233), the trypanosomiasis of cattle was dealt with. The investigators found that this disease was very widely distributed in the Congo, the infecting parasite being usually 7. dimorphon. It was also observed that domestic animals probably acquire a relative immunity to some strains of trypanosomes, and may even recover spontaneously. Trypanosomes were found in horses, mules and donkeys as well as in cattle, and also in Tvagelaphus scriptus. Pia) & Parasitol., Vol. XV Annals Trop. Med. & ENTRANCE HALL « ut) Thomas, assisted by Linton and Breinl (Memoirs XIII and XIV), established by a long series of experiments that trypanosomes found in (a) the cerebro-spinal fluid of Uganda sleeping sickness cases, (6) the cerebro-spinal fluid and blood of Congo sleeping sickness cases, (c) the blood of Congo ‘trypanosome fever ’ cases, and (d) the blood of Europeans infected in the Congo, were identical in animal reactions and morphology with 7. gambiense, Dutton. It was also found (1) that the periodicity of the parasite is a prominent feature, both in man and beast ; (2) that the passing of a strain from a susceptible into a very resistant animal does not attenuate the organism, and that the morphological character is retained after being passed through many hundreds of animals for nearly three years; (3) that the parasites in an animal may sometimes become more virulent, that such a strain may be particularly virulent for one species of animal, and that the more rapid infection is not due to the inoculation of a greater number of parasites than usual. In addition to 7. gambiense and T. dimorphon, other pathogenic trypanosomes were procured, and comparisons made between the above organisms and TZ. evamsi, T. brucei, T. equinum and T. equiperdum. Cultivation of the different parasites was also undertaken with success. Breinl gave a detailed account of the post- mortem changes in four cases of sleeping sickness (Memoir XVI). Extensive research was conducted into the treatment of trypano- somiasis, with the result that two drugs only were found to be of any value in the disease, namely, arsenic and ‘ Trypanroth’. Thomas introduced atoxyl, a meta-arsenic anilin compound, a drug which causes no pain on sub-cutaneous injection and may be administered over a period of many months.- He stated that it was ‘the only remedy at present giving a prospect of a cure.’ Although atoxyl would almost invariably cause the trypanosomes to disappear from the peripheral blood yet since the parasites frequently reappeared, it seemed possible that they might exist somewhere else in the body of their host in a form uninfluenced by the drug. Series of experiments were, therefore, undertaken by Benjamin Moore and Nierenstein, of the Bio-Chemical Department, and Todd. They found that in the treatment of rats infected with 7. brucei the administration of atoxyl, followed by bi-chloride of mercury, gave better results than treatment by atoxy] alone. 20 Moore, Nierenstein and Todd, continuing their researches on the treatment of experimental trypanosomiasis (Azmals, Vol. II, p. 265), found that in small animals, such as rats and rabbits, infected with T. brucei, the results of treatment by atoxyl followed by mercury salts were far superior to treatment by atoxyl alone; in large animals, as donkeys, on the other hand, the combined treatment was not found to be efficacious enough to be of practical value. These workers also studied the effects of therapeutic agents on trypanosomes in respect to (@) acquired resistance of the parasites to the drug, and (4) changes in virulence of the strains after escape from the drug (¢é7d., p. 221). Further bio-chemical research into the subject was made by Nierenstein, who observed the acidity and alkalinity of the blood in trypanosome infections, and found that whereas the total acidity of the blood serum showed a marked increase, the total alkalinity remained constant (7b7d., p. 227). Later he made an extensive study of the chemo-therapeutics of atoxyl (zbzd., pp. 249, 323 and 329). Breinl conducted some experiments on the combined atoxyl-mercury treatment of monkeys infected with 7. gambiense (2b7d., p. 345), and demonstrated that in five cases out of six, the administration of acetylated atoxyl and sublimate, and Donovan’s solution, to monkeys (Cercopithecus callithricus\, effected a complete cure. As the result of Plimmer and Thomson’s discovery of the trypanocidal action of antimony, Breinl and Nierenstein investigated the action of aryl-stibinic acids in experimental trypanosomasis (767d., p. 365), and showed that both ~. and m. amino-phenyl-stibinic acids are fairly powerful trypano- cides, the former being superior in its action. Owing to the satisfactory results obtained in laboratory animals, a trial of the former compound in sleeping sickness patients was advised, the dose suggested being the same as for atoxyl. In May, 1907, the Eighteenth Expedition of the School, consisting of Dr. Allan Kinghorn and Mr. R. E. Montgomery, was despatched to Rhodesia and British Central Africa to study the trypanosomiases of men and animals (Azmals, Vol. Il, pp. 53, 97, 333 and 387; Vol. III, pp. 259, 277 and 311). It was found that sleeping sickness had already invaded N.E. Rhodesia, the first case being seen in the Luapula division, adjoining the frontier of the Congo. Glossina surveys were made, and the suggestion first 21 advanced that Gl. palpalis and Gl. fusca were not the only transmittors of the disease, Gl. morsitans having also been observed in infected areas. Gland palpations were made in 26,928 natives, of whom 17°05 per cent. were found to have palpable glands. The percentage of positive punctures was 77°7._ The workers confirmed the belief of Dutton and Todd that gland palpation, combined with puncture, is a most useful measure, being a practical method of isolating infected natives, and preventing any rapid extension of the disease. It was found that there were between fifty and sixty known cases of sleeping sickness in the ‘country. The mode of introduction and prophylaxis of the disease were studied, and regulations drawn up by this Expedition were adopted and enforced by the Government. Concurrently with the research into human trypanosomiasis, the Expedition pursued an enquiry into trypanosomiasis of domestic stock in North-Western Rhodesia, in the course of which it was established that the disease was very prevalent in that area, and was due to 7. dimorphon, T. vivax and a trypanosome morpho- logically allied to Z. Orucez. These trypanosomes could be transmitted by Gl. morsitans, by Stomoxys calcitrans and by a species of Lyperosta. The question of association of big game and Gl. morsitans was considered, the opinion being that the distribu- tion of Gl. morsitans is entirely dependent upon the nature of the country and its flora, that the association with the fauna is largely fortuitous, and. that a perpetual supply of mammalian blood is not imperative, at least to its temporary existence. The classification of trypanosomes was attempted, the observers dividing the thirteen named species in the following way :— T. theileri, T. equinum, T. gambiense and T. equiperdum, easily recognised by their morphology or animal reactions; the nine remaining species sub-divided into three groups having as their types:—(1) Z. evansi (with T. brucei and T. sudanense). (2) I. dimorphon (with T. congolense and T. pecaudi, and (3) T. nanum (with T. vivax and T. cazalbout). Research was undertaken into many other points, and included observations on the parasites occurring in the intestine of G/. palpalis and in the intestine and proboscis of Gl. morsitans. The many problems of trypanosomiasis continued to occupy the " 22 energies of the staff at Runcorn. Yorke took up the study of immunity in trypanosome diseases. He was able to prove (Azmals, Vol. III, p. 565) that a diminution of the haemolytic complement only takes place in the last stages of the disease, and that, in an experimental case, the complement had practically vanished shortly before the death of the animal, when the blood was swarming with parasites. He also worked on the protective action of the serum of animals in a state of chronic infection or immune to various kinds of trypanosomes. Later, he investigated the condition of the blood which gives rise to the phenomenon of auto-agglutination of the red blood cells in animals infected with trypanosomes and to sleeping sickness in man (Pvoc. Roy. Soc., Ser. B, Vol. LXXXIII, p. 238, and Annals, Vol. IV, p. 529). Experiments showed that auto-, iso- and hetero-agglutinin exist in the blood of many normal animals, and are frequently present in much greater amount in the blood of infected animals. Apart from infection with trypanosomes, well marked auto-agglutination was found to be an extremely rare phenomenon. Ross and Thomson studied a case of sleeping sickness and demonstrated a regular periodical increase of the parasites (Annals, Vol. IV, pp. 261 and 395). These workers confirmed the claim that there is a life cycle of trypanosomes in the vertebrate host (2dzd., p. 465). In 1910, Stephens observed a marked peculiarity in the morphology of a trypanosome from a rat supposed to be infected with 7. gambiense. This parasite was described by himself and Fantham (Azmals, Vol. IV, p. 343), the animal reactions observed by Yorke (zézd., pp. 351 and 385), and a new species was founded, to which the name TZ. rhodesiense Stephens and Fantham was given, based on the peculiar posterior position of the macronucleus. It was later established that 7. brucez exhibits this peculiarity. In 1911, the Twenty-seventh Expedition of the School, consisting of Dr. J. L. Todd and Dr. S. B. Wolbach, was despatched to the Gambia to investigate sleeping sickness. Reports of the work done were published in the Axznals, Vol. V, p. 245. In the course of this investigation, 12,298 persons were palpated, and the observers put on record their opinion that gland palpation and puncture was by far the best procedure for the diagnosis of trypanosomiasis. It was found that at least o°8 per cent. of the Annal Trop. Med & Parasitol.. Vol. XV PLA TE V Pa (ns 7) te mason! r hs : LIBRARY 23 population of the Gambia was infected with trypanosomes. Strong recommendations were made for the control of the disease in the Gambia, including a continued examination of the whole population, the establishment of villages for isolation, observation and treat- ment of cases, and the appointment of a special staff for the administration and execution of these projects. Stannus and Yorke examined in rats the parasite from_a case of sleeping sickness contracted in Nyasaland (7did., p. 443), and were convinced that the trypanosome in question was not 7. gambiense but probably identical with 7. rhodesiense, which it resembled very closely, having a posterior nuclear form. A study was made by Yorke and Blacklock of the trypanosomes from a horse naturally infected in the Gambia (é:d., p. 413). The parasites consisted of a long form with a free flagellum, and also a short form without a free flagellum. The former was considered, from its morphological appearance and animal reactions, to belong to the 7. vivax group; the latter form was subsequently identified (Azmnals, Vol. VI, p. 107) as T. dimorphon, sensu Laveran and Mesnil. Blacklock measured one thousand examples of this form, and found that the average length was 13°3" (Annals, Vol. VI, p. 287). Measurements of one thousand examples of 7. vivax in goats were made by Blacklock, the average length being 27°7" (Annals, Vol. V, p. 521). He also, using the same strain of 7. vivay, measured fifty trypanosomes drawn from a rat and a rabbit respectively. In the former the average length was 21‘1p, in the latter 20°8m (zbzd., p. 537). In 1911, at the request of the British South African Co., . Dr. Kinghorn and Dr. Yorke were sent to Rhodesia (Luangwa Valley) to study sleeping sickness. It was quickly placed beyond doubt that Gl. morsitans was the carrier of the human trypanosome (Annals, Vol. VI, p. 1). The investigators inoculated rats from twelve cases of human trypanosomiasis, eleven of which occurred in villages in the Luangwa valley. In every instance they observed the posterior displacement of the macronucleus, characteristic of the trypanosome described by Stephens and Fantham; the animal reactions agreed in all respects with those obtained from infection with 7. rhodesiense. Elaborate transmission experiments with both wild and laboratory-bred Glossena morsitans were successfully carried out in rats and monkeys; the duration of the cycle in the 24 fly (approximately fourteen days) was found to be shorter than in experiments of previous investigators with Gl. palpalis and Gl. morsitans. It was observed that an infected fly retains the power of transmitting the disease during its life, and is infective at each meal, but that mechanical transmission does not occur if a period of twenty-four hours has elapsed since the infecting meal. Certain species of buck, viz., waterbuck, hartebeest, mpala and warthog, were found to be infected with the human trypanosome, as well as a native dog. Later (Axmals, Vol. VI, p. 269), it was found that 16 per cent. of the local game were infected with T. rhodesiense, and it was established that the game and fly strains were identical with the human trypanosome. In all, six species of tryanosomes were found in game and domestic stock in the Luangwa Valley (Azmnals, Vol. VI, p. 301), namely, IT. rhodesiense and T. pecorum, transmitted by Gl. morsitans and probably by insects other than tsetse-flies; 7. vivax and T. nanum, probably transmitted by Gl. morsttans,; and two others, one of which was possibly 7. montgomer:. At least 37°5 per cent. of the buck were found to harbour pathogenic trypanosomes. Ina later report (2b:7d, p. 317). a new trypanosome, 7. ignotum, infective to monkeys and a rabbit, was described. The vertebrate host was not discovered. This trypanosome is now known as T. stmiae, having been previously found in the same year by the Royal Society S.S. Commission. Still later (Annals, Vol. VII, p- 254), descriptions were given of 7. multiforme, sp.n., and of T. tragelaphi, the latter closely resembling 7. ingens. In the course of experiments in the development of 7. rhodesiense in Gl. morsitans (Annals, Vol. VI, p. 405), it was observed that the cycle was influenced to a marked degree by the temperature to which the flies were subjected, high temperatures (75° to 85°F.) being favourable and low ones (60° to 70° F.) unfavourable to the development of the parasites. Parasites might persist in the fly at at incomplete stage of their development for at least sixty days under unfavourable climatic conditions. It was found (Azmals, Vol. VI, p. 495) that in every fly capable of infecting animals with 7. rhodesiense the salivary glands were invaded, also that on every occasion on which the salivary glands were infective the trypanosomes in the intestines were virulent. Invasion of the salivary glands was only observed 25 in the case of flies infected with 7. rhodesiense and not in the case of any other trypanosomes met with in the Luangwa Valley or on the Congo-Zambesi watershed. It was calculated that approximately 3°5 per cent. of the flies might become permanently infected and capable of transmitting the: virus (Azmals, Vol. VII, p. 183). The chief reservoir of the human trypanosome was found to be the antelope. Stephens and Fantham (Proc. Roy. Soc., B, Vol. LXXXV, p. 223, and Annals, Vol. VI, p. 131) made a bionomic study of 7. rhodesiense. One thousand specimens were measured, and it was found that the average length was 23°6y, as compared with 7. gambiense 22'1m, and T. bruce: 23°2u. The average length of T. rhodesiense in man and other species of animals was ascertained. Those in the rat were found to be the longest (24°5) and those in the rabbit the shortest (19'4). Of the three species 7. rhodesiense was found to be richest in long and poorest in intermediate forms. Another series of measurements was under- taken (Annals, Vol. VII, p. 27), when each day, for the first ten days of infection, one hundred trypanosomes from the same rat were measured, the results being again compared with 7. gambiense. It was found that the day of infection was of great importance, as there was a great variation in the percentage of ‘stumpy’ forms on different days, ¢.g., 53 per cent. on the seventh day to 5 per cent. on the tenth day. The larger the sample of trypanosomes taken, the smaller the variation in the average length. J. G. Thomson succeeded in cultivating 7. rhodesiense by the use of a modification of the Novy-MacNeal-Nicolle medium. The changes taking place in the trypanosomes were described (Axnals, Vol. VI, p. 103), and it was noted that when development was rapid two distinct types could be distinguished on the fourth day. In cultures which develope more slowly the trypanosomes disappeared about the third or fourth day, reappearing about the sixth, and on the eighth day spirillar forms were seen to be splitting off. Differentiation into the so-called ‘male’ and ‘female’ forms took place during the eighth, ninth and tenth days. Inoculation of animals from the cultures was unsuccessful. In a subsequent investigation with J. A. Sinton (cézd., p. 331), Thomson successfully cultivated 7. gambiense as well as T. rhodesiense, the former for a period of thirty-seven days, the latter for twenty-one 26 days only. The life history of these trypanosomes in culture tubes was similar to that occurring in the gut of the insect host. The cultures lost their infectivity after the third day, and it was suggested that probably their transference to a new medium or environment similar to that of the salivary glands of the tsetse fly might be required to permit the full history of the trypanosomes being completed. Blacklock observed the vitality and changes undergone by trypanosomes in the cadaver of the animal host (tbid., p. 55): He found that 7. gambiense and T. rhodesiense can remain infective in the blood of the dead animal for forty-eight hours. Blacklock also made a study of the posterior nuclear forms of 7. rhodesiense in rats (Annals, Vol. VII, p. 101). He found that they first appeared in the blood from the sixth to the tenth day of the disease, in a count of a thousand trypanosomes, that they increased in numbers in the later stage of the disease, and that they increased relatively to other forms of trypanosomes. They showed definite powers of resistance to disintegration in the cadaver. of the animal host. It was suggested that such forms might occur as a constant constituent of certain strains. Stephens and Blacklock made a morphological study of T. brucei (the Zululand strain) and of the trypanosome of the same name from the Uganda ox (Azmals, Vol. VII, p. 303): They asserted the non-identity of the two strains, and proposed for the latter the name 7. ugandae. Blacklock and Yorke (zdzd., p. 603), studying the pathogenicity of 7. congolense (Broden) and 7. nanum (Laveran), came to the conclusion that they were the same parasite. Yorke and Blacklock studied the characters of the more important mammalian trypanosomes (Azmals, Vol. VIII, p. 1), and compiled a convenient table of the main differential points. The value of the cycle of the trypanosomes in their invertebrate hosts as an aid to differentiation of species was also put forward. Todd re-examined the flagellate found by Dutton in 1902 in the blood of Gambia house mice, and came to the conclusion that it was ZT. acomys (ibid., p. 469). Yorke and Blacklock experimented with antimony trioxide in the treatment of experimental trypanosomiasis (zdzd., p. 55), controlling and extending some of the work of Kolle and others on the use of this drug. Various strains were used in the experiments. dunals rop. ed. & 2arasitol, “ol. XV If, ; dnnals Trop. Med. & Pa I, Vol. XP PLATE VI LECTURE THEATRE C. Tinling & Co., Ltd., Imp. 27 It was found that small animals could withstand a relatively much larger quantity of the drug than large ones. Post-mortem evidence proved that the proportion absorbed during a period of six months was exceedingly small. A certain number of cures would seem to have resulted, as several animals remained negative without relapse for over two hundred days, and sub-inoculated animals were not infected. Most strains appeared very susceptible to the drug, but T. gambiense and T. lewisi proved refractory. Seidelin tested the effect of salvarsan-copper on white rats infected with a strain of trypanosomes of the 7. brucez group kept in guinea-pigs and rats in West Africa (Aznals, Vol. IX, p. 197). The best results were obtained with the injection of a dose of 0°'0064 gm.; in such a case the trypanosomes disappeared from the blood on the following day and remained absent for fifteen days, death cccurring on the twenty-eighth day; in several other cases the life of the animals was prolonged for a few days more. The Thirty-second Expedition of the School, consisting of Drs. Yorke and Blacklock, was despatched to Sierra Leone in 1914. Research was undertaken into the bionomics of G. palpalis in Sierra Leone, with special reference to its pupal habits (Azmals, Vol. IX, p. 249). It was found that the breeding-grounds of G. palpalis are not so strictly limited to the immediate vicinity of water as had hitherto been believed. Mangrove swamps do not constitute a breeding-ground. The pupae do not hatch when subjected to daily flotation cn sea watef. The ground around the trunk of oil palms which have not been stripped of their lower petioles constitutes an excellent breeding-place for G. palpalzs; they can breed in localities in which practically the only tree is the oil palm. A study of the food of G. palpalzs in the Cape Lighthouse Peninsula, Sierra Leone (2bid., p. 363), showed that about 8 per cent. of the wild G. falpalis in that district contained recognisable red blood cells—7 per cent. of mammalian origin and 1 per cent. nucleated red cells of unknown origin. Neither shed blood nor other fluid which 1s exposed (not covered by a membrane) can be imbibed by G. falpalis. Fluids such as solutions of sugar, sodium chloride, etc., protected by a membrane (e.g., thin rubber sheeting), were taken up, but less quickly and readily than blood. It was thought that in nature G. palpalis may, under certain conditions, take up fluids other than 28 blood. The human trypanosome (7. gambiense) was discovered in an ox in Sierra Leone (2did., p. 383), thus demonstrating that domestic stock forms a reservoir for the virus of sleeping sickness. Among other animal parasites found in domestic stock in Sierra Leone (267d., p. 413), TZ. congolense and T. vivax were most commonly encountered. Trypanosomiasis of cattle was common. Of one hundred and forty-three examined, trypanosomes were found in nineteen after a single examination. In 1919, Escomel recorded the discovery of trypanosomes in the blood of a patient coming from Peru, which he described and considered to be Schizotrypanum cruzi. Yorke, examining this description, was led to doubt, on morphological grounds, whether the identification was correct, and proposed for this Peruvian parasite the name 7. escomelz, in recognition of its discoverer. A feature of the symptomatology of the case was the overpowering somnolence from which the patient suffered, a symptom not hitherto noted in Chagas’ disease, although a striking feature of the African trypanosomiasis of man (Azmzals, Vol. XIII, p. 459). Macfie described a trypanosome found in the blood of a snake, Naja nigricollis, in the Gold Coast, resembling in some degree T. primeti, but differmg from it in size and proportion. He proposed for it the name 7. voltariae (Annals, Vol. XIII, p. 23). YELLOW FEVER In 1900, the Fourth Expedition, consisting of Drs. H. E. Durham and Walter Myers, went to Brazil to study yellow fever. Dr. Myers died of the disease in January, 1901, while Dr. Durham, who also contracted it, recovered and published a report of the work of the Expedition (Zemozrv VII) in 1902. The U.S. Yellow Fever Commission in Cuba had not yet established the transmission by Stegoweyia when the Liverpool Expedition set to work at Para; for which reason the course of their investigation was chiefly directed at first to the search for some protozoal parasite. Only seventeen autopsies on yellow fever cadavers were obtainable. An extremely slender filiform bacillus was observed to occur constantly in the tissues and intestines, and much time was spent in isolating it and in attempts at cultivation. 29 Dissection of specimens of Culex fatigans captured in suspected houses showed large numbers of a similar bacillus. Owing to the death of Dr. Myers, the research came to an end. In 1905, Professor Rubert Boyce was despatched by the School to New Orleans, to observe the work of the U.S. Medical Authorities in dealing with the outbreak of yellow fever there. In Memoir XIX Boyce gave an account of the vigorous campaign which was successful in ridding the city of Stegomyza. Subsequently, at the request of the Colonial Office he visited Honduras to make a report on the conditions existing in that Colony with reference to a recent outbreak of yellow fever (Waterlow & Sons, 1906). In April, 1905, a second Yellow Fever Expedition was sent to the Amazon, the members being Dr. Wolferstan Thomas and Dr. Breinl. A permanent laboratory was established so that patients could be kept under continued observation, and experiments maugurated which would have been impossible under other conditions. Series of reports by Thomas were published in the Annals, Vol. IV, p. 1. It was recorded that the most serious disease to which the foreign population was liable was yellow fever, and this disease was made the object of extensive investigation. ‘Chimpanzees have been successfully inoculated ; rabbits and guinea- pigs exhibited certain reactions when inoculated with infective blood from yellow fever cases or subjected to the bites of infected S. calopus.’ (See also Trans. Soc. Trop. Med. and Hyg., Vol. III, p. 59.) The resources of the laboratory in Mandos were placed at the disposal of the State, the doctors, the hospital and the poor of the city. Examinations of blood, agglutination reactions for typhoid and paratyphoid, bacteriological examinations of water and milk, post-mortems and pathological reports were made, and the laboratory so conducted as to be of the greatest possible service to the community. Papers on ‘Oesophagostomiasis in Man’ and the condition known as ‘ Mossy Foot’ were published in the Axzals, as also an article on the ‘Mosquitoes of the Amazon region’ by Newstead jointly with Thomas. In 1910, at the request of the Colonial Office, Professor Boyce was despatched to the Gold Coast and Sierra Leone, to report on the outbreak of yellow fever at those places (Z7ans. Soc. Trop. Med. and Hyg., Vol. IV, p. 33). It was observed that the so-called 30 classical type of yellow fever was comparatively rare amongst native races, and the reason was advanced that natives are partially immunised by being born and brought up in an endemic area (Annals, Vol. V, p. 103). Those removed in childhood from such an area become non-immune, and therefore lable to succumb to an epidemic. In 1911, the study of yellow fever was taken up with energy by Professor Boyce, who suggested and superintended the establish- ment of a Yellow Fever Bureau at the Liverpool laboratories. A Bulletin was issued, in which the Director of the Bureau, Dr. Harald Seidelin, and other investigators published the results of their researches. Abstracts of reports from all over the world were also made, and reviews published of current literature on yellow fever and allied subjects, including pappataci fever and dengue. Research was undertaken into the etiology, diagnosis and treatment of yellow fever. Seidelin observed the occurrence of protozoon-like bodies in the blood and organs of yellow fever patients (/ourn. Path. and Bact., Vol. XV, p. 282). At a meeting of the Society of Tropical Medicine and Hygiene, in January, 1911, these organisms were demonstrated in preparations of the blood and sections of the kidney, and later (Yellow Fever Bulletin, Vol. 1, p. 229), Seidelin described their morphology, and proposed for them the name of Paraplasma flavigenum, g.n., sp.n. In December, 1911, he was despatched to Yucatan, where an epidemic of yellow fever was raging; a report of this Expedition was published in Vellow Fever Bull., Vol. Il., p: 123. The total number of cases officially diagnosed during the outbreak was seventy-three, with thirty-eight deaths. The existence of a ‘microbe carrier’ was put forward as being responsible for the maintenance of the virus during periods when the disease is latent, it being suggested that the fragile infected Stegomyia could not be the lasting reservoir. It was also considered that one attack does not necessarily confer immunity. The usually mild form of the disease in natives was said to be due either to an unrecognised infection in childhood or to the hereditary transmission of anti-bodies from immune parents. Clinical features of the cases observed were given. Pavaplasma flavigenum was seen in fifteen confirmed cases of yellow fever out of sixteen examined. Summing up the results of his blood examinations for P. flavigenum Annals Trop. Med. & Parasitol., Vol. XV PEALE VAL MUSEUM ng & Co, Lid., Imp. 31 in undoubted cases of yellow fever, Seidelin found that on this and former occasions he had approximately one hundred and six positive cases out of a total of one hundred and twenty. Records were made of the examination of four hundred and twenty-one specimens as controls. In one hundred and thirty-six cases malarial parasites were found, and in two hundred and eighty-three no parasites at all, while in two young children under no suspicion of yellow fever P. flavigenum occurred. It was suggested that these, together with two others previously observed, might be ‘ microbe carriers.’ Blood from yellow fever cases was inoculated into four guinea-pigs, but none showed symptoms resembling the disease. A mosquito survey of Merida was made, and recommendations advanced as to methods of extermination. A section of Seidelin’s report was devoted to a reply to those who had criticised his work. In 1915, the Yellow Fever Commission (West Africa) concluded that no proof had been given that P. flavigenum was of a protozoal nature, and that the nature of the virus of yellow fever still remained undetermined. In December, 1912, Seidelin was sent to Jamaica in order to investigate the nature of the disease called ‘vomiting sickness,’ prevalent in that island during the winter months, and responsible for a considerable mortality, chiefly among native children. A report of this Expedition was published in the Azmals (Vol. VII, p. 377) and also in the Yellow Fever Bulletin (Vol. Ill, p. 7). Sixty-two cases were observed, but no causal organism was recog- nised ; it was concluded that ‘ vomiting sickness’ was a local disease and could not be accepted as a form of meningitis, which view had been advanced by Scott two years before the latter’s demonstration of its true origin. RELAPSING FEVER AND SPIROCHAETES By the death of Dr. Dutton from tick fever while serving with the Congo Expedition, the School suffered the loss of one of its most brilliant workers, who, although only twenty-nine years of age, had already won a recognised position throughout the scientific world. Towards the end of 1904, Dr. Dutton and Dr. Todd had reached Stanley Falls, and they were able to demonstrate independently the 32 cause of tick fever in man, a discovery made a few weeks previously by Ross and Milne in Uganda. Further, they were able to prove the transference of the disease from man to monkeys by means of a particular species of tick. In Todd’s report of this discovery (Memoir XVIII) clinical descriptions were given of twelve native and two European cases (the last two being those of the investigators themselves), all of whom were infected with a spirochaete thought at first to be S. obermezert. A number of inoculation experiments with laboratory animals were carried out, it being found that to monkeys alone the parasite appeared to be uniformly pathogenic. Observations were made upon the distribution and bionomics of the human tick in the Congo, and included in the report was a description of Ovnithodoros moubata by Professor Newstead. Research was proceeding at Runcorn upon material brought back from the Congo by Dr. Todd. Dr. Breinl, invalided home from the Fifteenth Expedition, carried out with Dr. A. Kinghorn extensive studies cn ‘tick fever’ and ‘relapsing fever.’ Observations were made on the animal reactions of the spirochaete discovered in the Congo cases of human tick fever, and brought to England in infected monkeys and ticks. In the course of these experiments, infection was produced not only in monkeys, but also in a horse, a dog, rabbits, guinea-pigs, rats and mice. This fact caused the observers to conclude that the organism was distinct from S#irochaeta obermeieri, pathogenic hitherto to monkeys only. Further experi- ments were undertaken, confirming this conclusion; and the new species was given the name of S#ivochaeta duttoni (Breinl and Kinghorn, 1906, Memoir XX). Studies were made of this organism (Memoir XXI), in the course of which a clinical comparison was made between African tick fever and European relapsing fever, and the research into the animal reactions of this spirochaete in various animals amplified and completed. Experiments in immunity were continued, the conclusions reached being: (1) In animals which have recovered from the infection there is a relatively active immunity of comparatively long duration; (2) immune serum cannot produce passive immunity, nor has it any curative action; (3) hyper-immune serum "does not protect a susceptible animal; nor does it prevent relapse, but it mitigates the severity of the infection and occasionally cuts short an attack; (4) there is a slight inborn immunity of short 33 duration. It was also shown that S. duttoni may pass 7m utero from mother to foetus, and extensive studies were carried out to determine the réle played by the spleen in infection by spirochaetes. Attempts were made to transmit spirochaetes by the bites of Czmex lectularius, but without success. Subsequently (Azmnals, Vol. I, p. 435) Breinl studied the morphology and life history of S. duttoni, while Markham Carter (2bid., p. 15) described the multiplication and important changes in form of S. duttoni in eggs laid by infected ticks. Still later (Annals, Vol. V, p. 479), Fantham studied the life cycle of spirochaetes, amongst those considered being S. duttont, S. recurrentis and S. marchouxi. Subsequently (Azzals, Vol. VIII, Pp. 471) he investigated the granule phase of the parasite, a detailed study being given, while serving with the Expedition to Khartoum, to S. bronchialis, in which it was found that the granules formed by the spirochaete were the cross infective stages of the organism (Annals, Vol. IX, p. 391). In 1917, while making microscopical examinations of stained smears from the stools of five hundred and fifty-four patients, admitted to hospital for dysentery, Carter found that 56°5 per cent. were infected with Spivochaeta eurygyrata. A control investigation on one hundred cases free from intestinal disorders showed 41 per cent. to be infected (Annals, Vol. X, p. 391). Repeating this investigation amongst a normal population, Macfie and Carter (Annals, Vol. XI, p. 75) examined eighty-two hospital patients suffering from some surgical condition, and twenty-three normal healthy men. None of the cases had ever resided in the tropics. Of the hospital patients 56°2 per cent., and of the healthy men 43°8 per cent., harboured S. eurygyrata. A second species of spirochaete was discovered in the intestine of one case, which, owing to its larger size and certain morphological peculiarities, was considered to be a new species, and named by them Sfirochaeta intestinalis. Macfie and Yorke examined the morphology of the spirochaetes responsible for European, African and Indian relapsing fevers (Annals, Vol. XI, p. 81), and reached the conclusion that there is at present no means of distinguishing these parasites morphologically. 34 AMOEBIASIS Research into the amoebae parasitic in the human intestine was undertaken by Fantham, and a study commenced of the life history of E. coli as seen in cultures (Aznals, Vol. V, p. 111). Carter, Mackinnon, Matthews and Smith conducted extensive researches into the protozoal findings in cases of amoebic dysentery. In their first report (Azmals, Vol. X, p. 411) they recorded the results of four thousand three hundred and thirty-four examinations of nine hundred and ten patients suffering from this condition. Protozoal infections were discovered in 44°2 per cent.; EZ. Azstolytica was found in 10°3 per cent. of the cases; Z. colz in 25°4 per cent. ; G. intestinalis in 18°6 per cent.; 7. ivtestinalis im 1°2 per cent. ; and C. meSnili in 2°7 per cent. Their second report (Azmals, Vol. XI, p. 27) recorded similar examinations of one thousand seven hundred and thirteen cases of dysentery. Stress was laid upon the necessity for repeated examinations of each patient, as cases found negative the first and second times may prove on further examination to be E. histolytica carriers. The subject of ‘negative periods’ (absence of vegetative forms and cysts) in infected cases was also dealt with. A third report of this investigation appeared in Azmals, Vol. XIII, p. 83. Yorke and the above-mentioned observers examined for intestinal protozoa three hundred and forty-four persons who had never been out of England (Azmals, Vol. XI, p. 87). Of this number, two hundred and six were healthy young men of about 18 years of age who had recently entered the Army. A single examination of each of these cases revealed the interesting fact that 3°9 per cent. were infected with £. histolytica. In an address to the British Medical Association (B.//./., April 12th, 1919), Yorke emphasized the importance of discovering whether the infection in this country is recent or otherwise. He was inclined to believe that it was not recent because (1) carriers must have frequently entered this country before the war; (2) all the necessary factors for the spread of the infection are to be found in this country; (3) there are authentic records of cases of amoebic dysentery and liver abscess before 1914. Stephens and Mackinnon treated eighty-one cases infected with E. histolytica with ‘alcresta ipecac,’ an adsorption compound of Annals Trop. Med. & Parasital., Vol. XV PLATE VJIll MAIN LABORATORY C. Tinling & Co., Ltd., Imp. 35 emetine and aluminium silicate (Azmuals, Vol. X, p. 397), with the result that about two-thirds of the patients were freed from amoebic cysts. Carter and Matthews, using Cropper and Row’s method of ‘concentration (Azzals, Vol. XI, p. 195), found £. histolytica cysts in. five of one hundred and thirty-three apparently negative cases which had already received three ordinary routine microscopical examinations. Smith made a mensurative study of the cysts of E. histolytica and E. coli (Annals, Vol. XII, p. 27), and investi- gated the question of the number of races in the former parasite (Annals, Vol. XIII, p. 1). It was shown that not all infections of E. histolytica remain constant from one day to another in the average size of their cysts. The species can be divided into two races characterised by larger and smaller cysts, respectively. Infec- tions with £. Azstolytzca in healthy carriers who have never been out of England were shown to be characterised by a smaller proportion of the ‘small’ race, and also by a reduced proportion of the larger cysts of the ‘ordinary’ race, as compared with infections from convalescent dysenterics from abroad. Investigating the incidence of amoebic dysentery in asylum patients never out of England (Annals, Vol. XIII, p. 177), Smith found that of five hundred and four patients examined, fifty-nine had acute dysentery, and in three cases vegetative E. Azstolytica were found in the stools. BERI-BERI Simpson and Edie undertook research into’ the relation of the organic phosphorus content of various diets to diseases of nutrition, particularly beri-beri. After a review of the work of Schaumann and others (Azzals, Vol. V, p. 313), the investigators recorded their own experiments with pigeons fed on various kinds of rice, white bread and whole-meal bread. A study was made of the anti- neuritic bases of vegetable origin in relationship to beri-beri, the properties of the yeast extracts investigated, and a method adopted for the isolation of torulin, the anti-neuritic base of yeast (Axmals, Vol. VI, p. 235). 36 HELMINTHIASIS Stephens (Thompson-Yates Lab. Reports, Vol. VII, p. 9), described the morphology of Gastrvodiscus hominis, of two new human cestodes, Dibothriocephalus parvus and Taenia bremnert, of a new linguatulid, Porocephalus pattoni (Annals, Vol. I, p. 549), and of a new human nematode, Stvongylus gibsoni (Annals, Vol. Ul, p. 315). Observations on the hooklets of Cystecercus cellulosae in man (Azzals, Vol. Il, p. 391) showed that there is an irregularity of development affecting both the number of the hooklets and, more especially, the size. A fluke, found by Newstead in the alimentary canal of a Nicaraguan turtle, was described by Stephens (Azmals, Vol. V, p. 497), who proposed for it a new genus and named it Desmogonius desmogonius. The fluke from the liver of native dogs at Kasauli, India, was separated by Stephens from the genus O pisthorchis owing to the existence of a process or pedicle bearing on its summit the genital opening and a ventral sucker, and placed in a new genus as Pavopisthorchis caninus (Annals, Vol. VI, p. 117). Breinl and Hindle described a new Porocephalus, found in the lung of one of their experimental monkeys, and distinguished from the known species by the presence of an appendage on the outer pair of hooks only. They proposed for it the name of Porocephalus cercopithect (Annals, Vol. II, p. 321). Dogs in Freetown were found by Yorke and Blacklock to be heavily infected with Anxkylos- toma caninum and A. ceylanicum, the species being present in about equal numbers (Azmals, Vol. IX, p. 425). In 1916, Stephens contributed the section on Helminths to ‘The Animal Parasites of Man,’ issued jointly with Fantham and Theobald. In 1917, at a Veterinary Hospital attached to a remount depot in the neighbourhood of Liverpool, Yorke and Macfie started an investigation into the parasitic worms causing a heavy mortality amongst herses recently imported from America. The parasites belonged for the most part to various genera of the family Strongylidae, and in the course of their study Yorke and Macfie described eight new species and one new variety, viz., Cylicostomum longibursatum, C. minutum, C. pseudocatinatum, C. pateratum, C. tridentatum, C. -triramosum, Cylindropharynx rhodesiense, 37 Gyalocephalus equi and Cylicostomum nassatum, Looss, var. parvum (Annals, Vols. XI-XIV). Yorke and Southwell described a nematode from the intestine of a zebra, which certain minute characters of the head, and also the position of the vulva, led them to regard as a new species, for which they proposed the name Crossocephalus zebrae (Annals, Vol. XIV, p. 127). FILARIASIS The second part (Memoir IV) of the Report of the Third Malarial Expedition was devoted almost entirely to Filariasis. Eight new species, found during the examination of a large number of West Afrcan birds, were described, namely:—F. cyfselz, F. spiralis avium, F. fusiformis avium, F. spiralis avium major, F. falciformis, F. bibulbosa, F. capsulata and F. shekletoni. Observations were also made on human filariasis in West Africa, it being found that throughout the whole of that area the natives appeared to be infected with F. nocturna, diurna and perstans. With regard to the two first species, the majority of’ the cases encountered were atypical, in that, embryos were either never absent from the peripheral blood, or the maximum did not occur at mid- day and mid-night or thereabouts according to the species. Among the former cases there were many showing decided periodicity, and, among the latter, the hour at which the maximum number was present varied considerably. In some cases, two maxima during the twenty-four hours were indicated. In- the examination (day blood) of three hundred and ninety natives of all ages up to about eighteen years, one case only, aged eleven years, was infected. The observers succeeded in infecting A. costalis (proboscis) with F.. nocturna. They considered that the weight of evidence was on the side of the identity of F. nocturna and F. diurna, but that many points remained to be cleared up before the question could be settled. A study of the periodicity of Microfilaria nocturna was made by Yorke and Blacklock (Aznals, Vol. XI, p. 127). They found that obstruction to the passage of M. bancrofti through the cutaneous vessels occurs at all times of the day and night, but is at a minimum 38 at the end of the period of bodily activity. Although this obstruc- tion aids in the piling up of the larvae in the cutaneous vessels, it is in no way responsible for the nocturnal periodicity,- which is primarily dependent upon periodic variations in the arterial supply of larvae to the cutaneous vessels. By reversing the hours of sleep and activity, cutaneous immigration gradually becomes diurnal instead of nocturnal, the complete inversion of the periodicity being accomplished in from four to eleven days. The number of microfilariae, as judged from the maximum concentration in the cutaneous blood, remained at practically a constant level during the period of observation. The number of microfilariae in the urine varied greatly, the variation giving no indication of either a nocturnal or a diurnal periodicity. The number of microfilariae in the renal and vesical vessels exhibited a nocturnal periodicity analagous to that in the cutaneous vessels. ENTOMOLOGY In 1907, Professor Newstead, jointly with Drs. Dutton and Todd, issued a report (Aznals, Vol. I, p. 1) upon the insects and other arthropoda collected in the Congo Free State. Among the new genera and species described and figured were Evetmapodites inornatus, Stegomyia luteocephala, S. albomarginata, Duttonia tarsalis, D. africana, Culex laurenti, C. par, Mimomyia africana, M. malfeytt, Boycia mimomytajormis, Haematopota duttoni, H. trimaculata, Tabanus billingtoni, Glossina maculata and Stomoxys omega. The habits and structural characters of the larva of Szmulium ornatum (a European species) were described. They occur on the under sides of submerged leaves or blades of grass in those parts of a stream which are rapidly flowing and fully exposed to the sun. Their mode of progression resembles that of looper caterpillars, spinning a network of silken threads along which they travel. The period of pupation varies from two to six days. The imagines escape through a slit in the thorax, and occasionally may be seen completely immersed in water with their wings folded so as to encase an air bubble. The distribution of tsetse-flies in the Congo was recorded and illustrated by a map, together with observations on the bionomics of the flies. The life history of Stomoxys calcttrans was also described in detail. dunals Trop. Med & Parasitol. Vol. XV PIA EX ENTOMOLOGICAL DEPARTMENT 39 In the following year, Newstead made a study of the bionomics of the common house-fly (Azmals, Vol. I, p. 507). The chief breeding-places were determined, and it was established that the life-cycle of the fly, in all kinds of fermenting material, is reduced to the minimum period of ten to fourteen days; and that in the absence of such artificial heat the cycle may occupy a period from three to five weeks or more, according to the temperature of the outside air. It was found that house-flies do not depend entirely upon excessively warm weather for breeding purposes. Methods of prevention were. suggested, and some notes appended on other insects found during the investigation. In 1908, Newstead went to Jamaica to study cattle ticks. In the course of the investigation, twenty-five estates were visited and the cattle inspected in each, and large numbers of ticks were collected in every district (Azmals, Vol. III, p. 421). Methods of treating tick-infected stock were made the subject of enquiry, and whenever possible practical demonstrations were given. The nature and extent of injury caused by insects and other pests to cultivated crops was studied, and methods of control advised. Descriptions were given of the nine species of ticks found in Jamaica, namely, Argas persicus, Margaropus annulatus australis, Rhipicephalus bursa, R. sanguineus, Dermacentor nitens, Amblyomma maculatum, A. cajanense, A. dissimile, Aponomma sp. In 1906, Newstead investigated the life history of Stomoxys calcitrans (Journ. Econ. Biol., Vol. I, p. 157) and, together with Stephens, described the anatomy of the proboscis of Glossina palpalis (Memoir XVIII, p. 53), and subsequently (Axzals, Vol. I, p. 169) that of Stomoxys calcitrans. In 1906, a former student, Capt. R. Markham Carter, I.M.S., forwarded to Newstead a species of tsetse-fly (namely, G. tachinotdes) from Arabia, this being the first observation of the occurrence of Glossina outside Africa. In 1910, Newstead described three new species of Glossina (Annals, Vol. IV, p. 369), namely, G. submorsitans, G. brevipalpis and G. f/uscipes. These new species were founded on an examina- tion of the morphological characters of the male genital armature. A revision of Glossina, based on a study of this structure, was made (Bull. Ent. Res., Vol. Il, p. 9), and later two further new 40 species of this genus were described, namely, G. austeni (Annals, Vol. VI, p. 129, and Bull. Ent. Res., Vol. IM], p. 355) and G. severini (Annals, Vol. VII, p. 331). In 1911, as a member of the Commission of the Royal Society to enquire into the relation of the African fauna to human trypano- somiasis, Newstead proceeded to Nyasaland, and for five months devoted himself to a study of the bionomics of the tsetse-fly (Glossina morsitans, West.), with a view to discovering its breeding grounds and devising means of checking its spread. The results of this investigation were published, jointly with Dr. J. B. Davey, in the Reports of the Commission, No. XV, p. 142. The physical features of the country were first described and an account given of the vegetation of the river and its borders, and the forest or fly area. The vertebrate fauna of the district were then dealt with. It was concluded that mpala antelopes supplied a very large propor- tion of the food necessary for the life and propagation of Glossina. Two species of birds were shown to prey upon G. morsitans. The breeding grounds of G. morsttans were thinly scattered over the whole of the country, and large numbers did not occur in any given spot. It was found by experiment that the average time between each meal was about two and a half days. The period that elapsed between the date of capture of the fly and the production of the larvae varied from two to twenty-nine days. The duration of the pupal period was found to be about twenty-five days. The period of chief activity of G. morsitans was between the hours of 10 a.m. and 4 p.m. In 1919, Miss A. M. Evans made a study of the genital armature of the female Glosszna (Annals, Vol. XIII, p. 31). In 1018, Newstead discovered that the innumerable papillae which form the sculpturing on the exterior of the prominent lobes at the anal extremity of the larvae of Glossina are respiratory openings, and evidently function as such during the inter-uterine life of the larva. Similar structures were found in the Hippoboscidae (Annals, Vol. IV, p. 93). Jointly with Carter, a new genus and three new species of anopheline mosquitoes were described (Aznals, Vol. IV, Pp. 377), namely, Dactylomyia, nov. gen., and Dactylomyia ceylonica, Pyretophorus cardamatisi and Cellia cincta, and later 41 six further new species and varieties were dealt with (Axxals, Vol. V, p. 233). In 1911, Newstead and Carter founded a new genus of Culicinae from the Amazon region, which they named Thomasina. The type species, Thomasina longipalpis, had previously been referred by Newstead and Thomas to the genus Mansonia, but it was now found that the morphological characters of the palpi and tarsi were so markedly different from those of - Mansonia that the species could no longer remain in that genus (Annals, Vol. IV, p. 553). Some mosquitoes of the genera Banksinella and Taeniorhynchus were described by Carter (Axnals, Vol. VII, p. 581) with a view to establishing the affinities of certain species and in reference also to the synonymy adopted by other students of this group of blood-sucking insects. In 1920, Carter gave an account of the male genital armature of the British anopheline mosquitoes (Azmals, Vol. XIII, p. 453). Extensive observa- tions were made by Blacklock and Carter on the bionomics of A. plumbeus (Annals, Vol. XIII, p. 421). It was found to be essentially a tree-hole breeder; larvae were taken from the water in rot-holes of elm, sycamore and other trees, from 2 to 20 feet above the ground. The breeding-places may occur in more or less isolated trees situated sometimes within a few yards of houses. A. plumbeus feeds on man both day and night. The observers obtained larvae from tree-holes in December to February, and they ascertained that thirty-five out of forty larvae survived freezing for five to thirty minutes. (For infection experiments with A. plumbeus, see p. 12). In 1912, Carter described three new species of the genus Tabanus, which he named Tabanus nagamiensis, T. fulvicapillus and 7. donaldsont, and in a subsequent study (Annals, Vol. IX, p. 173) eight* previously undescribed Tabanidae were dealt with. _ He also described three new African midges (Azmals, Vol. X, p. 131), Forctpomyta lefanui, Culicoides cordiformitarsis and Culicoides stephensi, and later (Annals, Vol. XII, p. 289) two others, Culicoides ocrothorax and Forcipomyia ingrami, a species of interest owing to the fact that, given. favourable opportunities for attack, its larvae prey upon the larvae of mosquitoes breeding in rot-holes in trees. In 1920, Carter, Ingram and Macfie commenced an exhaustive study of the Ceratopogonine midges of the Gold Coast, with descriptions of new species. In the first account of 42 these midges (Axmnals, Vol. XIV, p. 187), after a description of the technique employed, the observers dealt with the bionomics of the various genera. The second instalment (zbid., p. 211), which began a systematic account of the Ceratopogoninae, dealt with the genus Culicotdes, and included descriptions of sixteen species, eleven of which were new. The larvae and pupae of several species were described in detail. The third account of this investigation (zbid., p. 309) dealt with six new species belonging to three genera, of which one of the latter is new. In 1919, an article on the blood-sucking Nematocera was contributed by Carter to ‘The Practice of Medicine in the Tropics’ (now in the press). This dealt with the biting flies of the families Culictdae, Psychodidae, Chironomidae and Simuliidae. The account included general considerations regarding structure and bionomics, the species of malaria-carrying Awofheles being arranged in groups according to the nature of the evidence on which they were incriminated, and the classification. In the last section, the diagnostic characters of all the known (one hundred and twenty) Anopheline mosquitoes were given in synoptic tables. Newstead continued his researches on Coccidae, of which the British species formed the subject of a monograph, in two volumes, issued by him among the Ray Society publications, in 1900 and 1902. In 1908, he contributed an article on these insects to the reports of the Swedish Zoological Expedition to Kilimandjaro (Vol. II, Part 12); the same year he wrote of the scale insects and mealy bugs of Egypt (Lev. Univ. Inst. Comm. Res. in the Tropics Quart. Journ., Vol. III, p. 14), and reported upon a collection of Coccidae affecting plants in Java and West Africa (Journ. Econ. Biol., Vol. III, p. 32). In 1910 and 1911, he described two new species of African coccids (Journ. Econ. Biol., Vol. V, p. 18), reported on a collection from Uganda (Bwll. Ent. Res., Vol. 1, pp. 63 and 185), on another from South and South-west Africa, and on a third in the Berlin Zoological Museum (A@7¢t. aus dem Zool. Mus. in Berl., Vol. V, p. 155). Later, he commenced a series of studies on Coccidae, which are still being pursued (Bull. Ent. Res., 1913-1920). In these papers, over one hundred and sixty different species were dealt with, including the descriptions, with illustra- tions, of one hundred and six species and varieties new to science; the major portion of these are serious pests to various crops under 43 cultivation in tropical and sub-tropical countries. In 1gI0, Newstead served on the Special Commission appointed by the Government of Malta to suggest means for stamping out the fluted scale insect (/cerya purchasi), then threatening the orange-growing industry of the island. Regulations for the suppression of the pest were drafted and circulated. Other work of an economic nature undertaken by Newstead included an investigation into the food of some British birds (Suppl. to Journ. of Board of Agric., Vol. XV). This work, which had extended over a period of twenty years, was based upon over eleven hundred records, chiefly post mortem. His tentative verdict was in favour of the birds, the records showing what an important part is played by the majority of British birds in checking the increase and lessening the ravages of many insect pests of plants and crops. In 1910, Newstead dealt with some insects affecting cultivated plants in the West Indies (Journ. Roy. Hort. Soc., Vol. XXXVI, p. 53), and in 1913, jointly with Bruce Cummings, issued a paper on a gall-producing Psyllid from Syria (Ann. Mag. Nat. Hist., Vol. XI, p. 306). In 1910, Newstead went to Malta to investigate sand flies of the genus PAlebotomus, the main object being to discover the chief breed- ing-places of these insects, and to recommend. practical measures for destroying them in the larval stage. It was discovered that four distinct species of Phlebotomus occur in the island, previous investi- gators having notified one species only. The discovery of some important structural details concerning the anatomy of these insects was made and a long series of drawings illustrative of the salient characteristics was prepared, as well as a series illustrative of the internal anatomy (Azmals, Vol. V, p. 139). Later, he described some new species (Bull, Ent. Res., Vol. Ul, p. 361, Vol. V, p. 179, Vol. VII, p. 191, and Vol. XI, p. 305), and in 1913 dealt with three West African species (Bull. Soc. Path. Exot., Vol. VI, p. 124). Besides the special studies noted above, many articles on entomological subjects were contributed to a variety of journals by the Department of Entomology. In addition, innumerable collec- tions of insects submitted by the Imperial Bureau of Entomology, by the Belgian Government, and from other sources, were examined and identified. In 1915, Newstead went to France and Flanders, there to 44 organise measures of fly control. A report of this work was submitted to the War Office. By the request of the Royal Society, Newstead and others commenced in 1916 an investigation into the problems connected with the damage caused to grain and flour during transit and in storage. It was found that wheat and flour are liable to attacks and injury by acarids, of which Aleurobius farinae was most commonly encountered. It was established that mites will not injure wheat and flour in which the moisture content is 11 per cent. or under, whatever the temperature may be. The morphology and bionomics of the infesting mites were studied, and experiments carried out with regard to methods of destruction. Newstead and Morris also reported upon the non-parasitic or forage acari of the family Tyroglyphidae, to which Pillers added clinica] notes derived from veterinary experience (Royal Society: Reports of the Grain Pests (War) Committee, Nos. 2 and 8). David Thomson made feeding experiments with the European bed bug (C7zmex lectulartus) in various diseases (Annals, Vol. VIII, p. 19). He found that protozoa were absent from the gut of this species (one hundred and eighty-four examined). No acid-fast bacilli were found in one hundred and five bed bugs fed on lepers, nor in thirty-five others caught in bed mattresses of leper patients. Nothing abnormal was found in bugs fed on cases of lymphadenoma, carcinoma and malaria. Forty bugs fed on a case of spleno- medullary leukaemia all developed numerous Charcot-Leyden crystals in their intestines. ‘ Dutton, Todd and Christy described the Congo floor maggot, a blood-sucking dipterous larva found in the Congo Free State (Memoir XIII, p. 49). The larva was stated to be semi-translucent, of a dirty white colour, acephalous, amphipneustic, consisting of eleven segments, and to feed mainly, or entirely, at night. The duration of the pupal stage was a fortnight to three weeks. A light- brown fly, caught in many huts infested with the maggot, was subsequently identified by Austen as Awchmeromyia luteola. Newstead (A znals, Vol. I, p. 49) noted that the true larval stage is continued till after the formation of the puparium, and that a large percentage of the flies escape backwards from it. In addition to dealing with the anatomy and bionomics of 45 Ornithodoros moubata, the tick transmitting African relapsing fever (already mentioned above), Newstead, jointly with Todd, described a species of acarid found infecting the lungs of monkeys, namely, Pneumonyssus duttoni (Memoir XVIII, p. 41), and later he described another new acarid, Pxeumonyssus griffithi, found in the lungs of the Rhesus monkey (zdzd., p. 47). PROTOZOOLOGY Dutton, Todd and Tobey gave an account of certain parasitic protozoa observed by them in Africa (Memoir XXI, p. 87, and Annals, Vol. I, p. 285) in mammals, birds and reptiles, including full descriptions and figures of some forms of Leucocytozoon zlemanni, parasitic in a grey hawk of the Congo. Fantham studied the leucocytozoon, L. lovatz, of the red grouse, Lagopus scoticus, and observed the occurrence of schizogony in its life cycle (Azmals, Vol. IV, p. 255). He also studied a flagellate found in the alimentary tract of the body louse, which he named Herpetomonas pediculi (Annals, Vol. VI, p. 25), and of which he demonstrated the complete life cycle, notifying its occurrence in lice in England, as well as in India and Tunisia (24¢d., p. 403). Another Herpeto- monas, H. stratiomyia, sp.n., was discovered by Fantham and Porter (Annals, Vol. VII, p. 609) parasitic on the larvae, pupae and imagines of the flies S/vatiomyia chameleon and S. potamida. Research into induced herpetomoniasis in birds resulted in producing this condition in canaries, sparrows and martins by feeding them on insects containing herpetomonads; in some cases the infection was fatal. It was found that the cycle of the flagellate in the avian host resembled morphologically that in the insect (Azmals, Vol. IX, p. 543). Fantham and Porter studied the effects on their hosts of certain Myxosporidia inhabiting the gall bladders of various fish (Azuals, Vol. VI, p. 467). In 1916, Fantham contributed the section on the Protozoa to ‘The Animal Parasites of Man,’ issued jointly with Stephens and Theobald. Seidelin described some blood parasites in reptiles (Axnals, Vol. V, p. 371), and also some species of Klossiella in the kidney of a guinea-pig (Awmnals, Vol. VIII, p. 553). E.H. Ross observed the development of a leucocytozoon in a guinea-pig, for which he 46 proposed the name Lymphocytozoon cobayae (Proc. Roy. Soc., B., Vol. LXXX, p. 67); Sinton prosecuted research into the morphology and biology of Prowazekia urinaria (Annals, Vol. VI, p. 245); and O’Farrell, in a study of hereditary infection, with special reference to its occurrence in Hyalomma aegyptium infected with Crithidia hyalommae, gave an account of the four periods in the life cycle of this flagellate (Awmals, Vol. VII, p. 545). In 1917, Smith and Matthews investigated the incidence of intestinal protozoa in two hundred and fifty patients admitted to hospital for diseases other than dysentery (Aznals, Vol. X, p. 361). Entamoeba histolytica was found in 8 per cent. of the cases, EZ. coli in 19'2 per cent., G. znfestinalis in 8 per cent., C. mesnili in 2 per cent., and 7. intestinalis in 17 per cent. Of the two hundred and fifty cases examined, two hundred and two were suffering from non-intestinal complaints, and of this number 9°4 per cent. were found to be harbouring cysts of £. hzstolytzca. Among ninety-one men who had been to France only, two were discovered to be ‘carriers’ of £. histolytica. In a further investigation (Azzals, Vol. XI, p. 183), two hundred non-dysenteric patients were examined, and protozoal infections found in 34°5 per cent.; E. histolytica in 7°5 per cent. Matthews described and figured the characteristic morphological features of cysts of the common intestinal protozoa of man (Azmals, Vol. XII, p. 17), and later made a mensurative study of the cysts of £. coli (Annals, Vol. XII, p- 259), and traced the course and duration of an infection with this parasite (Azwals, Vol. XIII, p. 17). Matthews and Smith investigated the spread and incidence of intestinal protozoal infec- tions in the population of Great Britain. The first selected population consisted of four hundred and fifty civilians in the Liverpool Royal Infirmary (Azmals, Vol. XII, p. 349), of which 1°5 per cent. were found to harbour £. histolytica and 6°7 per cent. E. coli. The figures for army recruits, of which one thousand and ninety-eight cases were examined, were Z. /istolyzica 5°6 per cent., E. coli 182 per cent., E. nana 2'4 per cent., G. intestinalis 6°0 per cent., and C. mesnili two cases. In five hundred and forty- eight children, all under the age of twelve (Azmals, Vol. XII, p. 361), G. zvtestinalis was the parasite most commonly found. Of two hundred and seven male asylum patients (Azwals, Vol. XIII, 47 Pp. 91), 9'7 per cent. were found to be infected with Z. Azstolytica, 45°9 per cent. with £. colz, and 23'2 per cent. with C. mesuilt. University and School cadets were also examined; the same protozoa were found as amongst other series, but the number of cases recorded was too small to allow of conclusions being drawn as to incidence amongst this higher social class. In 1906, Drs. Fantham and Porter investigated the Isle of Wight bee disease (Annals, Vol. VI, p. 163). It was found that the disease was due to a minute microsporidian parasite, Nosema apis, sp. n., which gained access by the mouth to the intestines. Experi- mental work proving the pathogenicity of the parasite was carried out. It was found that Nosema apis was harboured by other insects besides the hive bees (Azzals, Vol. VII, p. 569). It was considered that a bee, itself apparently immune, can be a parasite carrier. A morphological study of Nosema apis was made, and the two phases in its life cycle demonstrated: (1) a multiplicative phase, termed merogony, which occurs in the epithelium of the chyle stomach and intestines of the bee; (2) a second phase, termed sporogony, leading to the formation of minute, resistant resting spores, which are shed in the faeces of the bee, fouling the surroundings of the hive and producing infection of fresh bees when swallowed in food or drink. An allied organism, Nosema bombi, sp.n., parasitic in, and pathogenic to, bumble bees, was discovered, and its life cycle, and suggested economic measures of control, set forth in a subsequent paper (Aznals, Vol. VIII, p. 623). . te t “ ; *<: * . ( oouiethk ok dite halite oF Dey aie Sees Ke : ; yn ee Ta Ra rts AS ate jE 38 Tare Sale Dae 2 lect Re ysl ix) > et i faa mela dny AO Fi Bae Bry bksgastt & aby eae hy Worsiot the th sae ‘ Seals tate ea ae hyo Cea 4x5 Shee eke a pee fais Theta e role ow e ie eee 5 Te). OL ee (it Buse 7 : 5 sid Lorre at ; 4 bares FE Vey 1h “ ii ; ee > body Y xp eerESs ; RT ee Cae Ly estet thay Aopen AEF nts (anes Seale Wade eee ai, the Rt {os ae wes £e. a oe he ety eee a) ; s mectin one doukia ras F bate one Kite aba onl lo? sae es See tthe cule . {Ca esCwct: 4 heeTid ieethid lig Vs Se {i} wk. tre 5b VERS fe a wihagere4 ninete totes +1 “et ony f sa viaat nent eed to a pasty fi awrelln. war seo tithe Pook death be fisaanotea as (ie Rete Se Oo Aietth | ageybtaeyal ite argaay 9349) eb ate Dak ¢ c1as0S eHyetiey eal sedate paosline ape 13 tosses tocarhaoes ‘ ce ae QUO AR oN ¢ ? * ° ° . = .$ 4 et oe £ rt - ai » i 4 ae ‘ 4 (= : ' i; bor ak 49 ON THE ‘ARNETH COUNT’ IN HOOK- WORM-INFECTED WHITE CHILDREN IN NORTH QUEENSLAND BY A. BREINL From the Australian Institute of Tropical Medicine, Townsville (Received for publication 31 January, 1921) In the past, work has been carried out by Breinl and Priestly (1914) on Arneth counts of healthy white school children, who had spent their lives in the tropical parts of North Queensland. The observations proved a decided shift of the Arneth index to the left, when compared with that of normal individuals in Europe. Later, this investigation was extended to healthy aboriginal children in Northern Australia (1917), and, furthermore, to native children in New Guinea (1915), living in an area where malaria, yaws and other parasitic diseases were found to be endemic. A shift of the Arneth index to the left was found in healthy aboriginal children of North Queensland analogous to that of children of European descent in North Queensland, but a much more pronounced shift was found in the native children of New Guinea. Taking advantage of recent opportunities, blood smears obtained from white children in North Queensland, suffering from ancylostomiasis were examined in order to determine the Arneth index. A number of the smears were collected by one of the members of the staff of the hookworm campaign, carrying out work at present in North Queensland ; others were obtained from children of various ages, who underwent treatment for hookworm infection in the Townsville Hospital. Previous observations by Knapp (1915) in India showed in the blood of hookworm patients a distinct shift of the Arneth index to Bie) the right, but the results, according to his own statement, ‘were on the whole equivocal,’ and he proposed to carry out further research. Macfie (1916), working on the Gold Coast, confirmed to a certain extent Knapp’s tentative results. Out of seventeen counts made on hookworm patients, about 30 per cent. showed, when compared with those of healthy natives, an actual shift to the right, 41 per cent. had a relative shift to the right, that is, ‘a slighter degree of shift to the left than is found in apparently healthy natives,’ and 29 per cent. had a definite shift to the left. He concluded from his results that ‘there appeared unquestionably a tendency to develop a shift to the right in patients infected with hookworms.’ In the present investigation the same technique was employed as in the previous work; all counts were performed by myself, Dr. Priestly and myself having performed the previous counts. In this way the results, so far as technique is concerned, are comparable with those obtained previously, and the personal source of error has been, so far as possible, excluded. Two hundred consecutive leucocytes were counted in two sets of 100, and only when the two sets of figures differed but slightly were the counts taken into consideration. All the children from whom the blood was obtained lived in areas where malaria 1s practically unknown, and any definite change found must be attributed to the effects of hookworm infection. The figures (Table I) were separated according to age groups, and the table shows that the averages for the age groups between five and fifteen years are fairly constant. The Arneth index for children two and three years old is much lower, and approaches that found in healthy North Queensland children. The disproportionate rise for children four years of age may be due to the small number of observations. The total averages prove conclusively that the average Arneth index in hookworm-infected children shows a decided shift to the left. A comparison of the Arneth index of hookworm-infected white children in North Queensland with that of children living in New Guinea shows a striking similarity, and strengthens the assumption that the comparative increase in the Arneth index in the latter locality was due to the great incidence of latent and active infection $i to.0 LL _ $z.£1 LS.0 TET 0$.0 76.21 08.0 | fz.01 £.0 | Qz.01 gt.o 77.6 $9.0 $9.91 £t.0 bE.11 0S.0 | 2£.01 ob.o | Si-b1 t£.0 zL.o1 19-0 $L.z1 94.0 $2.51 $.0 1.77 9.0 6.21 ZO Ib.z1 sapryd s[]92 3seyy | -omsoq $.6z $.z tb 93-7 gf.1 Ly.z SL.1€ gS-o 00.£ fb.b€ 0$.0 £0.€ o1.S€ 19.0 Zg.£ bl.it gt.o “1.€ 7.6 $S.0 tlc 6L.Sz z£.0 12.2 to.Lz St.0 19.7 LL.gz 38:0 $6.2 $Z.S€ 6£.0 10.£ 66.0£ 19.0 10.£ Qz-rE 25.0 $S.z gZ.zf 04.0 to.£ 9.62 Lo .5 1.1b g:0 S.£ oth g:0 Lz saq49 qeaponu s|euor -oydurhy | -ouopy -1sueL], adie] 1.9S 96.0 8g-18 z9.gh < 6£.6+ zg.£S 62.45 6g.£5 1.6 65.95 1£.9+ ££.$S or.6b 96.9+ g.1+ 1.98 6.g£ aryd -o1jnau aeajonu -oydiour ~4f0d ee eee % SINOOD IVILNaXTsIG 74 | $.0 Sb 9.07 93-£8 1Z.0 £1.2z og-Er g1.£g £1.0 1S.z oz-br 00.£g — $Lx Sz.11 gf.og 6£.0 PE. 63-51 00.£ £1.0 1£.z gS-br S$z.£g to.0 6z.z zbebr 18.£g — 99-7 | €gEr 60.8 t1.0 $8.1 fz.£1 2£.£3 07.0 €S.z 06.£1 zE.Sg $0.0 Ql-z gt.z1 98-78 $0.0 £$.z gS-b1 go-£g 1.0 $6.2 2g.f1 S18 1.0 $.z Z.$1 $.S6 — Z.0 | + $.$2 $.0 ZS 6.31 S.z£ So | o€ o-bz | | xepuy A Al IIL Nf ora \ pandapomitien aa % o.tF NOILVOISVID HLINYY — SS i a ee ae $1 TL * waspyry> jo Jaquinyy sieak $1-Z (aqrym) UaIP[Iy> [ooyps Puefsuvand qUu0N ereak OI-1 page ‘eoumg man jo WaIP[IyD aaneN S1o03z aay §2 amongst children, amongst whom hookworm infection is probably widespread. The significance of the shift of the Arneth index to the left is still uncertain. It is, however, possible that in such diseases as hookworm infection and malaria, where the destruction of red cells goes, for a time at least, hand in hand with an increased activity of the blood-forming organs, the increased activity extends to the new formation of leucocytes, and in consequence a greater number of young leucocytes are met with in the peripheral blood. The differential counts indicate that in younger children— between two and four years of age—the relative number of polymorphonuclear neutrophile leucocytes is decreased, whereas that of the lymphocytes is increased. After the seventh year the relative proportion of the various forms of white blood corpuscles is fairly constant. As is to be expected, the relative increase in the number of eosinophile leucocytes is well pronounced throughout our series of counts. REFERENCES Breint and Prirstty (1914). Ann. Trop. Med. and Parasit., Vol. VI, p. 565. (1915). Ann. Trop. Med. and Parasit., Vol. IX, p. 495 (1917). Ann. Trop. Med. and Parasit., Vol. X, p. 427. Kwaprp (1915). Indian Med. Gaz. Vol. L, p. 65. Macrte (1917). Report of the Accra Laboratory for 1916, p. 44. Churchill, London, 53 BRONCHOMONILIASIS COMPLICATING PULMONARY TUBERCULOSIS IN A MetivE OF “THE GOLD “COAST, WEST AFRICA BY J. W. S. MACFIE AND A. INGRAM (Received for publication 2 February, 1921) Since Castellani discovered the condition in Ceylon in 1905, bronchomoniliasis has been identified in many parts of the world, especially in tropical and sub-tropical countries. In Africa, Chalmers and Macdonald (1920) studied a number of cases in the Sudan and Egypt, and Pijper (1917) has noted the presence of the disease in South Africa, but, so far as we are able to ascertain, no cases have hitherto been recorded from West Africa. For this reason, a short account will be given of a case which has recently come under our notice at Accra, in the Gold Coast, West Africa. The case was not a pure bronchomoniliasis, but occurred in a patient suffering from pulmonary tuberculosis, a form of mixed infection which apparently is not uncommon, and has been observed previously by de Mello and Fernandes, Castellani and Chalmers, and others. History. We are indebted to Dr. J. R. Moffatt for the following history of the case. I.D., a native (Buzaburime), aged about twenty-five years and a member of the Police Force, admitted to the Native Hospital, Accra, on the 29th of September, 1920. The patient stated that he had suffered severely from cough for at least two months previous to admission. Upon béing closely questioned, he further admitted that for at least eighteen months he had experienced attacks of illness, accompanied by cough, at irregular intervals. Physical examination revealed dullness at the apex of the right lung, chiefly supra-clavicular; at the base of the left lung there was a considerable area of dullness extending as high as the 54 angle of the scapula, and at its upper portion, especially on the anterior aspect, bounded by a hyper-resonant area. The cough was frequent and harrassing ; the sputum copious and in appearance like thin flour paste. Sweating was inconsiderable. Although the breathing was rapid, the patient never suffered from dyspnoea. The temperature chart kept whilst the patient was in hospital showed an irregular fever similar to that which might have been expected in a case of pulmonary tuberculosis. The treatment given was cod- liver oil and, for a few days only, potassium iodide. During the last two weeks of his illness the patient showed some improvement and gained weight; on the night of the 8th of November, however, he had a sudden and copious haemoptysis and died within an hour. A specimen of the sputum of this case was forwarded to the laboratory for examination as to the presence of tubercle bacilli upon the 30th of September; none were found on this occasion, but it was noted that the sputum had a curious appearance, suggestive of saliva containing small particles of macerated bread, and accordingly another specimen was asked for which should be taken after thoroughly washing the mouth with a weak antiseptic solution. On the 2nd of October the second specimen of the sputum was examined, and was found to contain numerous yeast-like cells (Monilia sp.) but no tubercle bacilli. Cultures were made from this specimen of the sputum upon Sabouraud’s maltose agar and glucose agar, and within twenty-four hours a copious creamy-white growth of the Monilia had made its appearance on both media. On the 5th of October the sputum again showed yeast-like Monilia cells and also a few coarse hyphae; tubercle bacilli were not detected. As potassium iodide has proved a valuable remedy in broncho- moniliasis, it was suggested that it should be tried in this case, and this was done for a few days, ten grains being given thrice daily. The effect was to reduce the number of Monilia cells in the sputum very greatly, and to reveal the presence of tubercle bacilli which in all probability had been overlooked at previous examinations. A specimen of the sputum examined on the 8th of October showed very numerous tubercle bacilli and no Monilia cells, and as the potassium iodide appeared to be causing the patient some discomfort, owing to the increase in the quantity of his sputum, it was then stopped. Sputum examined on the 14th of October, and 55 later, showed that the Monilia cells had reappeared and that the number of tubercle bacilli appeared to be fewer. The patient died on the 8th of November as the result of an haemoptysis, and a post-mortem examination of the body was made on the following morning. Post-mortem Examination. The following were the notes made at the examination. Body: that of a young native man, rather emaciated. Abdominal cavity and its contents: appeared to be normal. Spleen: not enlarged, weight seven and a half ounces, no visible morbid condition. Kidneys and liver: showed venous engorgement but no other pathological signs. Gall bladder: collapsed, empty. Mesenteric glands: not enlarged. Right lung: adherent to the chest wall at its apex; on removal and section a cavity about the size of a walnut was found at the apex, it contained a blood clot; the whole of the upper lobe and a portion of the middle studded with small tubercles. Left lung: completely collapsed, visceral and parietal pleurae very greatly thickened and of a creamy- yellow colour; the pleura covered in parts with a deposit of the colour and consistence of cream cheese, the pleural cavity contained about four ounces of turbid straw-coloured fluid; the substance of the lung studded with numerous caseating tubercular nodules, and ~ at one point apparently communicating with the pleural cavity. Lymphatic glands at the root of the neck and in the mediastina: enlarged, tuberculous. Smears of the creamy exudate in the left pleural cavity showed numerous tubercle bacilli and yeast-like Monilia cells, also a considerable number of short septate branching hyphae. Sections of the lungs showed that both were the seat of advanced tubercular disease; sections of the thickened pleura of the left lung and of a mass of the pleural exudate showed also the presence of Monilia. Organism. The Monilia found in this case was easily obtained in pure culture by inoculating tubes of Sabouraud’s maltose agar and glucose agar. It was gram-positive but not acid fast. It grew well on most solid media, but especially well on glucose agar, and produced rapidly a diffuse, spreading, creamy-white growth. Under anaerobic conditions its growth was slower. Gelatin and blood serum were not liquified by it, and did not become pigmented. In broth and peptone water it caused a white deposit to be thrown 56 down whilst the media themselves remained clear; in peptone water a slight surface pellicle was formed. It produced a thick white growth on potato. On solid media the growth was almost entirely composed of yeast-like cells; in some fluid media hyphae predominated. Its qualitative bio-chemical reactions may be tabulated as follows :— Arabinose ig ies fa, AO Inulin O Rhamnose (isodulcite) wim O Amygdalin O Galactose ee 55 2. 0AG Helicin Oo Glucose ... aeA A TAG Phlorrhizin O Laevulose 33; Be ... AGs Salicin Oo Mannose nis ae Bere) Glycerol ... O Lactose ... se pe Pr 1,0) Erythrol ... O Maltose ... ue iy ... AGs Adonitol O Saccharose a a oh AG Dulcitol ... O Amylum oe see aoe Inosite O Dextrin ... ser fe Soe slg 0 Mannitol... O Glycogen 2 he vO Sorbitol ... oO The symbols, representing: A—acid; G—gas; s—slight; -and O—neither acid nor gas. : The production of gas in laevulose and maltose was slight. If the cultures were kept for two weeks or longer the acidity produced in the five sugary media indicated tended to be superseded by alkalinity; this was earliest seen and most pronounced in glucose and saccharose. At first no change was produced in litmus milk, but later, after about ten days, alkalinity”developed; no clot was formed and the medium was neither decolourised nor peptonised. Indol was not produced in peptone water. As gas was produced in glucose, laevulose, maltose, galactose, and saccharose, the organism comes into the fifth group of species of Monilia, called the Tropicalis group, according to the classifica- tion of Castellani and Chalmers (1919). In this group are placed (loc. cit. p. 1084) M. tropicalis, Cast., M. paratropicalis, Cast., M. pulmonalis, Cast., M. nivea, Cast., M. imsolta, Cast., and M. enterica, Cast.; but from the table given by the same authors (pp. 1082-1083) it would appear that M. faecalis, Cast., and M. metatropicalis, Cast., should also be included. A somewhat later table given by Castellani (1920) differs slightly from that given by Castellani and Chalmers and omits certain species whilst intro- 57 ducing some additional ones. According to it, acid and gas are produced in the five sugary media mentioned by five species only, namely, M. enterica, M. faecalis, M. metatropicalis, M. para- tropicalis, and M. tropicalis. Reverting to the species given by Castellani and Chalmers, which include the five given by Castellani alone and three others, it will be seen that the bio-chemical reactions of the organism isolated from our case do not agree entirely with those of any of them (see table). The reaction in litmus milk suffices to distinguish it from Table showing the more important biochemical reactions of the species of Monilia of the Tropicalis group. o o 2 2 i igcasis 5 SS eM a lees Spal hes pecies of Monilia ft 3 E £ 3 + g 5 F: B 7 =_ xo Re et a hehehe hbee (5 2p. Or fad 5 | DE ARYEr SCO Miss, cee | oss / Alk AG | AG | AG} AG | AG. As | As (0) oO A WAS faecapis wee, ans. ove / DPs | AG | AG | AG |} AGs| AGs| O (0) oO fo) As M. insolita... cee oss / atk | AG | AG } AG |} AG | AG As oO oO oO M. metatropicalis... eee || AC AG | AG | AG |} AG |} AG O ce) oO oO fc) M. nivea a ae oss / atk AG | AG | AG |} AG | AGs O Oo AG 10) As M. paratropicalis ... / Alk | AG | AG | AG/| AG} AG O | Avs | O | O ’ e) / | M. pulmonalis oa ree AlkD | AG | AG | AG | AGs;} AG Avs ie) A AGs M. tropicalis ... ve on A AG | AG | AG |} AGs| AGs_ O oO oO oO Species isolated from case of | O / | oe alae at Alk AG | AGs| AGs| AG | AG O oO ? Oo cera f Broth A = acid: G = gas 3,8 = slight; ve= very slight; O = neither acid nor gas produced ; Alk = alkalinity ; C =clot; D = decolourisation; C = clear; CTP = clear thin pellicle; P = peptonisation. all the other species of the Tvoficalis group excepting M. enterica and M. nivea. The former of these two produces slight acidity in mannitol and dextrin, reactions which are not produced by our species. As regards the latter, /. mivea, acid and gas are produced 58 in raffinose, and acid but only a small amount of gas in saccharose. We were unable to test the reaction of our species in raffinose, but in saccharose much gas was produced, and only a small amount in laevulose and maltose. It is admitted, however, that many of the species of the Genus JMonilia have not permanent bio-chemical reactions, a point emphasised by Castellani himself, and if they are liable to vary outside the body, it seems not unlikely that they may vary also according to their host. The very slight differences noted between the bio-chemical reactions of the organism recently isolated by us and those of J. nzvea are, therefore, probably unimportant. M. nivea was originally found in sputum, and is considered by Castellani and Chalmers to be of doubtful pathogenicity. It is of interest, therefore, to recall that the organism resembling this species which we have isolated was found not only in the sputum but also, after death, in the body of the patient. SUMMARY A case is recorded in which bronchomoniliasis complicated pulmonary tuberculosis in a native of the Gold Coast at Accra. The patient died of an haemoptysis whilst under observation. At the post-mortem examination both lungs were found to be tuberculous. The left lung was collapsed, and the pleural cavity partially filled with exudate. In this exudate and in the thickened pleura over the lung Monilia was present. The organism, which belonged to the Tvoficalis group of Castellani and Chalmers, closely resembled in bio-chemical reactions M. nivea, Cast. (1910); without raffinose we are unable to state whether the species found at Accra is distinct from /. nzvea, Cast. REFERENCES CasTELtant, A. (1905). Ceylon Medical Reports. (1920). Milroy Lectures on the Higher Fungi. Fourn. Trop. Med. and Hyg., Vol. XXIII, p. 118. and Cnarmers, A. J. (1919). Manual of Tropical Medicine, Third Edition, pp. 1070- 1092. London: Bailliere, Tindall and Cox. Cuatmers, A. J., and Macponatp, N. (1920). Bronchomoniliasis in the Anglo-Egyptian Sudan and Egypt. ‘Fourn. Trop. Med. and Hyg., Vol. XXIII, pp. 1-7. pe Metto, F., and Fernanpes, L. G. (Reviewed in the Trop. Dis. Bull., Vol. XIV, p. 244). Pryper, A. (1917). Med. Fourn. 8. Africa, Vol. XII, pp. 129-130. 59 NOTES ON A CASE OF INDIGENOUS INFECTION WITH P. FALCIPARUM BY B. BLACKLOCK (Received for publication 8 February, 1921) In their preliminary note on this case, Glynn and Matthews (1920) have furnished an account of the findings at the post-mortem examination and also details of the history. For convenience we may recapitulate here briefly the main facts. History. The patient was a girl, aged 18, who had never been out of the British Isles; she was born in Liverpool and had once been South—to London in 1914, from July to September, during which period she spent three weeks at Littlehampton; she had lived for the last ten years in the same house in Liverpool. She died on October 12th, 1920, after a period of illness which commenced (on her return from a holiday in a northern health resort) with ‘feverishness and flushed appearance’ on October 1st, followed on October 2nd and 3rd and several other occasions by vomiting. On October 8th she was overcome with faintness in the street, and had to be helped home. Her condition rapidly became worse with severe headache, thirst; photophobia, delirium, paresis of legs, anuria, and finally coma. The temperature on October 11th, the day before death, was 100°F., and on October 12th, just before death, 102°F. Further investigation elicited the facts that she had always been pale, had suffered from headaches since Christmas, 1919, and that she had vomited on September 29th, 1920. The following notes on the post-mortem examination are quoted from the paper mentioned above :— ; ‘ The post-mortem examination was made twenty hours after death. Blood : anaemic. Brain: apparently normal, no meningitis. Lungs: a moderate amount of oedema and muco-purulent bronchitis. Heart: weight, 8 0z., normal, no fatty degeneration. Mouth: healthy, no trace of pyorrhoea. Ocesophagus, stomach and intestines opened from end to end—normal, no evidence of blocd. Some of thejlumbar lymphatic glands slightly enlarged, and red like haemolymph glands. Liver: weight, 3 lbs., slightly browner than normal, no haemosiderin reaction. Spleen: 1 lb. 7 02., retains its shape, of normal consistency, but dark, rather like 60 malaria. Pancreas, suprarenals, kidneys and bladder normal. Uterus: normal size, but menstruating ; one haemorrhagic corpus luteum. Bone marrow of the shaft of the femur a uniform terra-cotta red, that of the sternum redder and more succulent than normal.” The following is a short account of my observations on the material examined. It is convenient to state here that the specimens from this case, which were demonstrated by me at a meeting of the Royal Society of Tropical Medicine and Hygiene, were by some inadvertence included under Col. James’s exhibits, and erroneously stated to have been derived from a case in Sheerness. (See Lancet, January Ist, p. 26.) Blood film. About a third of the red corpuscles contained young trophozoites, but owing probably to post-mortem changes the parasites appeared small and contracted, giving an appearance similar to that seen in sections. Not only was the total number of red cells infected large, but also multiple infection was common, many cells containing two, three, four, up to five parasites. Gametocytes were present in small numbers most of them being fully developed crescents, some of them presenting a peculiar appearance owing to the red cell being visible all round the parasite and extending some distance beyond it, and not chiefly in the concavity of the crescent; other forms were oval or spherical, and in addition there were seen solid-looking parasites of irregular outline which might be interpreted as developing forms of gametocyte. A small number of segmenting forms with a number of merozoites varying from ten to twenty-four also occurred in the blood, the crescents and fully divided forms being in about equal proportion, Nucleated red cells were numerous, as was observed in the preliminary note, the figure given being 3°8 nucleated reds to 100 leucocytes (500 counted), while my figure is slightly higher, 4°9 to 100 leucocytes (1,000 counted). They presented much diversity in the form of the nucleus, single-nucleated forms predominating, but forms with double and multipartite nuclei also being found. Pigment was present in mononuclear leucocytes and also free in the plasma, some in the latter possibly being due to the breaking of segmenting forms in making the film. Spleen smear. he chief feature to attract attention was the occurrence of very numerous segmenting forms, not all at the same 61 stage of sub-division, the merozoites numbering from eight to twenty-two. There were a few crescentic, oval, and spherical gametocytes. An occasional parasite, even in the segmenting stage, was found ingested by phagocytes. Pigment, both black and golden yellow, was present in considerable amount, ingested by large mononuclear white cells. Bone marrow smear. Here crescentic and oval forms of gameto- cyte were numerous in quite different proportions to the numbers in either the blood or spleen smears. A few segmenting forms occurred and many small trophozoites, of which some were free. Among the large mononuclear cells, many were observed which contained eosinophil granules, and many also containing pigment. Several questions of interest arise in connection with this case, and it is necessary, in spite of the limited character of the observa- tions made on the case during the period of her illness and the small amount of material available for examination, to endeavour to answer some of the questions. 1. Was this a primary acute attack following directly the incubation period, or a secondary acute attack supervening in a person already infected for a considerable time ? 2. By what means did the patient acquire infection ? 3. At what time and where did she become infected ? I. WAS THIS A PRIMARY OR SECONDARY ACUTE ATTACK? (a) Primary acute attack. The absence of any history, previous to October Ist, 1920, of fever, shivering or sweating, is in favour of this being a primary acute attack. Spleen. Support is also lent to this theory by certain appear- ances presented by the spleen. The noteworthy condition seen in section of this organ is the remarkable congestion, with dilatation of the sinuses. These facts are illustrated by photograph I, repre- senting a section from the spleen of the case under discussion. That the large size of the spleen is due to a recent enlargement of the organ is further demonstrated when we observe that the increase of size is due to engorgement with blood and not to increase of fibrotic tissue, nor to the cellular elements of the spleen itself. 62 (6) Secondary acute attack. It is quite possible for a person to harbour and present for considerable periods in his blood P. falczparum without the produc- tion of definite signs and symptoms, provided the person is taking quinine or other drugs. This fact has been observed frequently, and instances will be found in the work of Stephens and his collaborators (1917). Similarly it is recognised that persons suffering from relapses of malignant tertian malaria may have parasites present in the peripheral blood for varying periods before Section of normal spleen. X about 280. Photographs lent by Professor Glynn Puoto.I. Section of the spleen showing the Puoro. II. Malpighian body with vessel cut obliquely and below it the accumulation of red blood corpuscles. x about 280. the next attack occurs. But it is also possible that a person who is infected may, even if untreated, remain without definite signs or symptoms for a long period and then suddenly develop an acute attack; instances of incubation periods up to ‘months’ are mentioned by Craig (1914). It should be observed, however, that in experimental infections in which healthy persons are injected with blood containing parasites or are bitten by anophelines containing sporozoites in their salivary glands, such long incubation periods are not recorded. Ross, whose contribution to the elucidation of 63 this case is dealt with more fully below, does not supply information which is of assistance concerning this point. In the case under consideration the only evidence of old standing symptoms or signs which might possibly be attributed to malaria are that the girl had suffered from headaches from Christmas, 1919, her pallor, and the fact that she vomited on September 29th. With regard to these phenomena, they may be attributed to so many other causes that they can give us no clue. Spleen. The features which may be regarded as indicative of chronicity are the size (644 grms.) and lack of: diffluence. As regards the former, Dudgeon and Clarke (1918-19) give the weight of the spleen in a series of fatal cases of pernicious malaria due to P. falciparum as varying from 250 to 450 grms. ; the highest weight in a chronic case was 960 grms. On the other hand, James (1920) emphasises the great splenic enlargement observed in indigenous cases of simple tertian malaria in children. Previously Osler (1901) mentioned a case of death from accident during the second week of simple tertian malaria where the spleen weighed 800 grms., was very dark red, and diffluent. II. BY WHAT MEANS DID THE PATIENT ACQUIRE INFECTION ? The two known methods by which it is possible to transmit malaria to a healthy person are the inoculation of infected blood and the transmission by the bite of a mosquito with infected salivary glands. The incubation period, using such methods, varies considerably for malignant tertian parasites, and it is necessary to enquire into the periods and how they are estimated. 2 The inoculation of Infected Blood into Healthy Persons Of the numerous records obtained by inoculating blood containing malaria parasites into healthy persons only a small number is concerned with the malignant tertian parasite. Some of the early experiments collected by Thayer and Hewetson (1895) are, for comparative purposes, set out here in tabular form. It will be seen from Table I that in the early experiments the parasite incubation was long. These early experiments are, however, 64 open to several objections. In Gualdi and Antolisei’s first two experiments the inoculation of blood from quartan patients resulted in the appearance, in the subjects of inoculation, of malignant tertian parasites. The authors had to account for this by discovering - later that the patient whose blood was inoculated harboured both forms of parasite. In the third case recorded by Gualdi and Antolisei, and that of Di Mattei, crescents only were visible in the blood injected, while in the last case, that of Sacharow, not only was the injection small in amount and given subcutaneously, but also the blood was taken from leeches which had been fed four days previously on the infected person and which, since feeding, had been kept on ice. Taste I. Plasmodium falciparum. Incubation period following blood inoculation. Early experiments. Day on which, Amount in inoculated person Observer Route of of Blood, —__- Remarks Injection injected Fever Parasites commenced | appeared | C.C. Gualdi and Antolisei | Intravenous +4 3 10 10 Source of blood, quartan cases. Gualdi and Antolisei Intravenous | a I2 12 Source of blood, quartan cases. Gualdi and Antolisei ? | 2 9 Co) Crescents only found in blood | injected. Crescents seen in inoculated person in 18 days. Di Mattei . Intravenous ...| ? 16 | Ina few Crescents only found in blood | | daysafter| injected. Crescents seen in injection | inoculated person in 25 days. Sacharow .-, Subcutaneous I 12 13 Blood kept in leeches on ice 4 days before injection. Experiments were performed later by Bastianelli and Bignami, which are set out in Table II, and from this series a very different idea of the incubation period following inoculation is obtained. Parasites appeared in the peripheral blood in so short a period as two days after injection of 2 c.c. of infected blood, and infection resulted after such small quantities as 0'2 c.c. of blood injected. No mention is made as to the date of crescents appearing in the blood in these cases. 65 It appears, then, that the incubation period after experimental inoculation, whether as regards occurrence of fever or appearance of parasites in the blood, may be exceedingly brief in malignant tertian malaria. . Ross (1920) has suggested the possibility of this case having acquired infection by inoculation of blood. There was no evidence in the history of the case that such a mode of infection could have been the cause. She had cocaine injections for removal of teeth in July, 1919, and in February, 1920; for the removal of the teeth, mentioned as done on 14th September, 1920, the anaesthetic used was ether, and no evidence was obtained either that any such injections as morphia had been given, or that infection could have Taste II. Plasmodium falciparum. Incubation period following blood inoculation. Later experiments. Amount Day on which, : of blood in inoculated person Observer Route of injected |_— ——______ Remarks Injection cc. Fever Parasites commenced | appeared Bastianelli and ..| Intravenous... 2°0 2 2 Bignami 510 2 2 Few parasites seen in injected blood. O75 5 ? Few parasites seen in injected | blood. o2 4 ? Fair number of parasites seen in injected blood. arisen from abrasions of skin or mucous membranes having facilitated transmission. It is regrettable that Ross, who has sources of information inaccessible to others, should not have communicated the facts in such a form as to have added to our knowledge of the various points of interest which must have arisen as the result of his observations. The incubation period of malignant tertian malaria under such circumstances, the dates of the first appearance of symptoms and parasites, and also the important question as to the time of the appearance of crescents, are all matters of sufficient interest to make welcome any additional facts relating to them. Plasmodium falciparum, incubation period following the bites of anophelines previously infected. 66 Incubation period of P. FALCIPARUM following the Bites of Infected Anopheline Mosquitoes The records of experiments of this nature do not comprise numerous observations, and in.some cases the data given are insufficient for our present purpose. In Table III are shown the results of some experiments in which more complete data are given by the observers. Taste IIT. | Healthy individual Tempera- VLG ture Observer Anopheline or Number | Number Fever at which Remarks used | Bred of mos- of days com- | Parasites mos- quitoes | they were | menced appeared | quitoes fed fed in days in days kept Bastianelli and A. maculipennis ...| Wild ... 2 g-12 | 10-13 30° C, Bignami (1899) ) | | Simple Tertian Malaria in 17-19 | days. Schiiffner (1901) ...| A. vagus ... --| Wild ... ? 2 15-16 17-18 Room Large rings found. Jancsé (1908) .| A. maculipennis ...| Wild ... 2 I 12 12 24° C. Quinine, 1 gm. on 7 day after bites. Jancsé (1908) .| A. maculipennis ...| Wild ... I 2 10-15 11-16 20°C. | Quinine given af first bite, 1 | I gm, 15 Mitzmain (1916) ...| A. guadrimaculatus | Bred ... I I 10 | Io | 21-22°C. In these experiments the maximum parasitic incubation period of P. falciparum after infection by means of bites of infective anophelines is eighteen days, the minimum ten days. Before accepting the results of these observers, however, it is necessary to enquire more closely whether they are justified in regarding the experiments as conclusive. In Bastianelli and Bignami’s experiment wild A. maculipennis were fed, for the purpose of infecting them, on a patient suffering from malignant tertian fever; this patient after seventeen to nineteen days from the - commencement of feeding the mosquitoes on him showed in his blood the parasites of simple tertian fever. If the wild mosquitoes 67 infected him with Plasmodium vivax it appears not improbable that they might also infect with P. vivax the healthy person on whom they fed, and, therefore, one would expect that corroborative evidence such as the date of appearance of crescents in this case should be given. Apart from this, there is evidence, in some of these records, that the population used for experiment was subject to multiple infection. Reference has already been made to the fact that Gualdi and Antolisei, using the blood of patients supposed to be carrying P. malariae, obtained in the person injected P. falciparum This may be capable of many explanations, even excluding Laveran’s theory, recently revived by Grassi (1920), of the unicity of the parasite, for example that they conveyed the parasite of malignant tertian fever by the inoculation, or that the person inoculated was himself already suffering from infection with P. falciparum or that he acquired such infection from mosquitoes independently of the inoculation. In the same way, in Bastianelli and Bignami’s case, additional evidence as regards the species of parasite recovered is not available. In Jancso’s experiments the administration of quinine may have influenced the incubation perod. Mitzmain’s accidental case appears to be the most satis- factory. These considerations are mentioned in order to draw attention to the limited amount of reliable information available concerning the incubation period of P. falciparum. Factors such as the use of wild mosquitoes, experimenting among an already infected popula- tion, and the administration of quinine after the infective bite reduce materially the value to be attached to the figures. A further point to be mentioned, which may seriously affect the parasitic incubation periods given above, is that, as noticed previously, parasites are frequently present in relapse cases before a temperature reaction occurs. The importance of remembering this fact is seen when we consider Tables I, [I and III. It will be noted that in these experi- ments, fourteen in number, parasites were always discovered either after the day of fever or on the day of fever, with the exception of one case (di Mattei), in which they were found ‘a few days after injection.’ From what has been said, therefore, it is seen that the incubation period of malignant tertian malaria is a subject which would repay 68 further study. At present the evidence points to a minimum incubation, for inoculation of two days, and for mosquito bites of ten days. In view of the absence of evidence of inoculation infec- tion in this case, we must suppose that an anopheline was the means of infection. There appears to be no sufficient evidence to exclude the possibility of a more brief incubation than ten days, the more so when we consider that a formerly accepted minimum incubation period of ten days for blood inoculation has given place to a later minimum incubation period of two days. III. AT WHAT TIME AND WHERE DID THE CASE BECOME INFECTED? It has been necessary to deal with a portion of this question above in discussing incubation periods, but there are other facts which may be employed to throw light on the subject. Some assistance may be obtained from a study of the crescents and sporulating forms. Thomson (1911) states that, as a general rule, crescents appear in the peripheral blood on the fifth day after the attack of fever. Marchiafava and Bignami (1894) found crescents in the finger blood of thirteen primary cases, usually between the seventh and eighth day of the disease, rarely as soon as on the fifth. Bignami and Bastianelli, who examined cases by spleen puncture, found crescents in the spleen as early as the seventh and eighth day, and exceptionally also in the peripheral blood. It is seen from these observations that the spleen did not contain crescents any earlier than the peripheral blood. This corresponds with the findings in the present case, because an examination of smears of peripheral blood, spleen and bone marrow showed that whereas crescents were scanty in both peripheral blood and spleen, they were very numerous in the bone marrow. From this fact alone it could be deduced that death had occurred too early for crescents to be liberated from the bone marrow in large numbers. It has been shown by Marchiafava and Bignami (1894) that in certain cases of fatal malignant malaria, crescents may be scanty in the peripheral blood, spleen, and other viscera, while a very large preponderance of them is found in the bone marrow, where they may be seen in all stages. They conclude that the bone marrow 69 forms, if not the only, at least the most suitable site for the growth of crescents. They compare the passage of crescents from the bone marrow, where they are formed, with the corresponding passage of nucleated red cells from the bone marrow where they also are usually generated, into the general circulation. Dudgeon and Clarke (1918-19) did not find crescents in eke cases; they state in general of their fifty-one cases that no histological changes worthy of special reference were noted in the thyroid, bone marrow, testicles and pituitary body. They cite a very acute case of the haemorrhagic type, but no special mention of the condition of the bone marrow is made; it is a significant fact, however, that this case died in three days. Applying the crescent periods and incubation periods to the present case we have a sequence, tracing back from the day of death, as follows :—October 12th, day of death, crescents present in small numbers in blood; allowing five days for crescents to appear in peripheral blood after paroxysm gives October 8th as day on which in a normal case the paroxysm should have occurred ; allowing four days before this as the time when sporulating parasites first appeared in the peripheral blood, brings us to October 4th. From September 27th to October 4th is left for parasitic incubation period on the theory that she became infected by mosquito bite in the health resort in which it is known she was exposed to anophelines. This parasitic incubation period (seven days) is, even on the figures derived from the available experi- mental evidence, a probable one, and the history of the case supports it. It should be recalled that in this patient no shivering or sweating occurred and that we are dealing with a case of poor physique and considered to have been anaemic, the effect of which conditions upon incubation and reaction may be highly important, although by no means sufficiently recognised. Incubation will necessarily vary with the rapidity of the cycle of development of the parasite, the special reproductive energy of the particular parasite involved and the capacity of the infected human organism to facilitate or impede such parasitic reproduction. In the light of our present knowledge of incubation periods, the occurrence of a mosquito infection six years ago or an inoculation infection eight months ago seems improbable, and we have further 70 no evidence at present to warrant us in regarding the case as one of endemic malaria. We are, therefore, compelled to decide whether she was infected at Liverpool or in the resort referred to. The evidence appears to me to support the last named as the place of infection. THE SIGNIFICANCE OF CRESCENTS Thomson (1911) considers the development of immunity to be necessary before crescents can be produced, but he thinks that they arise from ordinary merozoites, the period required for full develop- ment from merozoites to crescents being about ten days. He found, in common with previous observers, that crescents appeared in the blood about five days or more after the paroxysm, but while accepting the existence of a condition of immunity as essential to their production, he does not attribute their delayed appearance to the lapse of time required before immunity is established but simply to the length of time required for their development. The idea that the delay was due to the time required for immunity to estab- lish itself had been put forward by Marchiafava and Bignami (1894) partly and also by Stephens and Christophers (1908). From a study of the appearances in this case, it is clear that the crescent- producing process was more advanced in the bone marrow than in the spleen or blood, and if the crescent-producing process is an ‘immunizing’ one, we may safely assume that its effect was first felt by the parasites in the bone marrow. Some evidence which might support the view that the process is connected with immunization may be obtained from a comparison of the relative numbers of crescents and segmenting forms present in the peripheral blood, spleen and bone marrow, respectively. In the peripheral blood the number of crescents and segmenting forms was about equal, in the spleen the segmenting forms far exceeded the crescents (fifteen to one), while in the bone marrow the segmenting forms were far outnumbered by the crescents (one to ten). Unless we assume that segmentation occurs at different hours in the bone marrow and spleen, or that red corpuscles containing young tropho- zoites cannot enter the bone marrow, on neither of which points is there any evidence, we are compelled to conclude that some agency in the bone marrow which causes the production of crescents 7) also prevents the completion of segmentation; that is to say, the bone marrow is the seat of a process which hinders the asexual while it facilitates the sexual development. The number of crescents produced in any area such as the bone marrow which contains only a proportion of the total blood must be so small relatively to the areas where segmentation is occurring freely, that it seems.evident that a certain time must elapse before the crescents reach the visibility point in the peripheral blood. This, in my opinion, is the real cause of the delay in their appearance, not only in primary cases but in relapses, and the delay seems bound to occur, even supposing that the ‘immunizing’ process already exists or becomes operative immediately in the bone marrow or other organs. _It appears that this delay must occur, and that it is not necessary to attribute it to a growth period of the parasite, of the duration of which growth period this very delay is the only evidence. In this case the ‘immunizing’ process had commenced in the bone marrow, but there was no evidence, as indicated by great crescent production, that it had commenced in the other tissues available for examination. It is possible that in other cases it may occur early in other organs also. NATURE OF THE ‘IMMUNIZING’ PROCESS WHICH RESULTS IN THE PRODUCTION OF CRESCENTS Nothing definite can be deduced as to what this process is or what are the factors which call it into operation. But that it is in many cases quickly operative in the bone marrow is obvious, not only from this case but also from the observations mentioned above of the early appearance of crescents in bone marrow. It is also clear that many primary cases in which this process is already established in the bone marrow, nevertheless die, which supports the view that if the process is an ‘immunizing’ process it is focal in origin. That the process is initiated by the mere presence of, or rupture of, segmenting forms, or by the toxins or pigment liberated at sporulation, is unlikely, in view of the observation in this case that in the spleen, where the most numerous segmenting forms occurred, crescents were scanty. 72 SUMMARY 1. A patient who had never been out of the British Isles died of malignant tertian malaria in Liverpool on October 12th, 1920. 2. There is evidence that this was an acute primary attack. 3. There is no evidence to show that this case acquired infection by other means than mosquito bite. 4. From a consideration of the records of incubation period and crescent formation, it is probable that the infection was acquired on or about September 27th, on which occasion she was in a northern health resort where anophelines are plentiful (A. maculipennis, A. bifurcatus and A. plumbeus). 5. Some evidence is given that crescent formation commences in the bone marrow, and that it is accompanied there by a failure of complete development in the asexual forms. 6. The ‘immunizing’ process which causes the above effects in all probability commences in some cases very early in the infection, but the crescents indicative of this process do not appear in the peripheral blood for some time. 7. The late appearance of crescents in the peripheral blood in infection with P. falciparum is explained on the ground that the source from which they arise is limited in extent. REFERENCES Bastranettt, G., and BrGNaMt, A. (1899). Atti della Soc. per gli Studi d. Malaria, Vol. I, p. 28. Cratc, C. F. (1914). Osler and McCrae’s Modern Medicine, p. 73. - Dupceron, L. S., and Crarke, C. (1918-9). Quart. Fourn. Med., Vo). XII, pp. 372-389. Grynn, E. G., and Matruews, J. C. (1920). British Medical fournal, pp. 811-813, Novy. 27. Grassi, B. (1920). Quoted by Sella, Internat. Fourn. Pub. Health, Vol. 1, No. 3, p. 321- James, S. P. (1920). Malaria at Home and Abroad, p. 150. Jancs6, N. (1908). Atti della Soc. per gli Studi d. Malaria, Vol. IX, p. 143. Marcriarava, E., and Brcnamt, A. (1894). Syd. Soc. Monog. on Malaria, pp. 64 and 211. Mrrzmatn, M. B. (1916). U.S.A. Put. Health Rep., Vol. XXXI, No. 6, p. 301. Oster, W. (1got). Clifford Allbutt’s System of Medicine, Vol. IT, p. 730. Ross, R. (1920). British Medical Fournal, Dec. 4, p. 871. Scuiirrner, W. (1901). Zeitschr. f. Hyg. u. Infektionskr, Vol. XLI, pp. 89-122. Stepuens, J. W. W., and Curistopuers, S. R. (1908). The Practical Study of Malaria and other Blood Parasites, p. 52. and Orners (1917). Ann. Trop. Med. and Parasit., Vol. XI, pp. 103-110 and 158-164. Tuayer, W. S., and Hewerson, J. (1895). Yobn Hopkins Hospital Rep., Vol. V, p. 35. Tuomson, D. (1911). Ann. Trop. Med. and Parasit., Vol. V., p. 57- 73 OBSERVATIONS ON MOSQUITOES IN THE ISLE OF MAN BY B. BLACKLOCK AND H.1,F., ,EARTER (Received for publication 22 February, 1921) PLATES X—XIV In recent years much interest has been shown in the mosquitoes of the British Isles, and many new observations have been made which increase our knowledge of the species which occur here. Twenty-two species, including three members of the genus Anopheles, have now been recorded from the United Kingdom, and many of them are also known to occur in Ireland; but, so far as we are aware, no information concerning the mosquitoes of the Isle of Man is available. Having recently had occasion to visit the Island, we took the opportunity of investigating the occurrence and distribution of mosquitoes. As, however, our visit extended over only a portion of November and December, it was impossible to make any detailed investigation of even those mosquitoes of which the over-wintering stages are known and readily found. Our activities were for the most part confined to localities easily accessible by railway or tramway, and were, therefore, mainly limited to the coastal region. The places visited and the records obtained are shown in the appendix and map. GENERAL CONSIDERATIONS I. ANOPHELINE MOSQUITOES. A. bifurcatus, L. The larvae of this species were abundant and widely distributed over the Island, and were found without difficulty in spite of the fact that residual breeding-places only could be discovered at this time of the year. As is sometimes the case with mosquitoes of this kind, larvae were found in certain of the breeding-places only after considerable searching ; this was especially noticeable in the more extensive breeding-places, such as the marsh- 74 land (Photo. No. 1) in the neighbourhood of Ballaugh and Sulby. On the other hand, relatively large numbers were sometimes obtained in a short time in the more circumscribed waters, such as the artificial pond at Groudle Glen (Photo. No. 2) and in the ditch at Castletown (Photo. No. 3). In the latter case, in the small ditch shown on the right of the photograph, over sixty larvae were captured in ten minutes by one person using a small scoop. As will be seen in the photographs, breeding-places of A. difurcatus were found in various types of country, even close to the sea (Photo. No. 4); they occurred also in situations where free water was only discovered by close inspection and was chiefly limited to hoof prints, etc. (Photo. No. 5). They were also found in the near neighbourhood of towns, as shown in Photo. No. 6 (Appendix, No. 15). A. maculipennis, Mg. Hibernating females of this mosquito were not found in such large numbers as (in view of the numerous potential breeding-places observed) we expected. All types of cow-sheds, stables, and other out-buildings from those of modern construction—high-roofed, well-lighted and well-ventilated —to those of primitive form—small, dark and badly ventilated— were systematically examined ; it was naturally impossible, however, to obtain access to houses, and, therefore, it is not possible to give records relating to them. In only one case (Appendix, No. 21, Photo. No. 7) was any considerable number of females found, when one hundred and forty-seven were captured, this representing the great majority of those present in the building. In accounting for the relatively small numbers seen, the following two facts, which came under cur notice, are possibly of some importance : — 1. Cleaning and lime-washing. In several instances we were informed that a thorough sweeping of the walls and roof to remove cobwebs had recently been carried out; this procedure in some cases had been followed by lime-washing. 2. Accidental cleaning of the roofs. Having observed in some cowsheds and stables with low flat roofs that cobwebs existed only in the corners, or over the manger, we made enquiries as to the reason of this. It is to be attributed to the practice of carrying on a fork the hay used as fodder, thus repeatedly brushing the greater portion of the roof. Annals Trop. Med. & Parasitol., Vol. XV PuotoGraPH No. PuorocrarpH No, 2 I PIG AMD y 4. 75 A. plumbeus, Steph. Although the Isle of Man is by no means heavily wooded, we experienced little difficulty in discovering the larvae of A. plumbeus on numerous occasions. The breeding-places were similar in nature to those of which we have given illustrations in our previous papers (1920). We found here a surprising difference in the tendency of the Spanish chestnut (Castanea sativa) to form rot-holes, and although no systematic survey was carried out, yet of nineteen breeding-places discovered, four occurred in this species of tree; whereas in Liverpool and district, of sixteen breeding-places found, none were present in the Spanish chestnut. Photo. No. 8 displays the most interesting breeding- place of this mosquito that we have yet discovered ; a full description of it is given on p. 87 (No. 34). Interesting breeding-places were also found immediately adjacent to an out-door latrine (Photo. No. 9) at a country farmstead, and near the centre of a town as in Photo No. 10 (Appendix, No. 31 f). It is necessary to note here that in no instance were the larvae of Ochlerotatus geniculatus, Ol., discovered. So far as our investiga- tions have gone, although we have commonly found this species in the United Kingdom in association with A. plumbeus, we have entirely failed to find it either in Ireland or the Isle of Man, although A. plumbeus is abundant in both places. II. CULICINE MOSQUITOES. Culex pipiens, L. Females of this species were found in several of the localities in which buildings weré visited. They were frequently found associated with A. maculipennis in occupied cow- sheds, stables and piggeries, but were distinctly more numerous in cooler stituations, z.e., cellars, lofts, out-houses, etc. (Appendix, p. 88). Theobaldia annulata, Schr. Larvae, pupae and adults of this mosquito were discovered in various places. The adults were relatively uncommon, but females were occasionally found in buildings, and, on one occasion (November 18th), both sexes were emerging from pupae contained in a wooden tub. This was standing in the back garden of a cottage at the Nunnery Mill, Douglas; the position of the tub in relation to the cottage is seen in Photo No. 11. The dimensions of the tub were 18 inches in top 76 diameter and 12 inches in depth; it was nearly full of rain water, which, on being stirred, produced a very evil odour due to a mass of putrifying leaves lying on the bottom; no larvae were found. Eighty-four pupae were collected from the tub, and from seventy- eight of these, twenty-seven males and fifty-one females emerged. Pupae of 7. annulata were also found at Ramsey on November 22nd, in a marshy meadow situated behind the town (Photo. No. 6). Larvae of the second, third and fourth instars were abundant, and occurred in several different types of breeding-place (Appendix, p. 88). Culicella morsitans, Theo. Larvae of various sizes were found; they sometimes occurred with the larvae of 7. annulata, but were more definitely limited in regard to habitat. They were always found in open water containing abundant vegetation, and, in marshy land, appeared to favour more particularly the immediate neighbourhood of clumps of rushes (Appendix, p. 89). Culicella fumipennis, Steph. Larvae of this species were found in one locality only, namely, in the extensive peat bog-land or ‘curragh’ in the vicinity of Ballaugh. They were, however, taken in two sites, but the nature of the breeding-places in regard to the types of vegetation present and colour of the water (reddish-brown) appeared identical. These larvae, some of which appeared but half-grown, were of a characteristic yellowish colour, and could immediately be distinguished from those of C. morsitans, which were darker and distinctly brown (Appendix, p. 89). APPENDIX, GIVING RECORDS OF SPECIES OBTAINED, UNDER PLACE NAMES ARRANGED IN ALPHABETICAL ORDER Anopheles bifurcatus, L. (1) BALLAUGH, December 6th. (a) The country immediately north of the railway to the East of Ballaugh Station presents features which are peculiar to this portion of the Island. It is a flat, desolate marshy country, with large expanses of water interrupted by collections of low bush and reeds. In Photo. 1, which gives a good idea of the appearance of much of this region, known as ‘the Curragh,’ the foreground repre- sents an area adjoining the path, completely waterlogged and Annals Trop. Med. & Parasitol., Vol, XV PLATE XI PHotoGraren No. 3 PHotoGRarH No. 4 PuotroGraen No, 5 . ime. oA itl communicating with the open water beyond. Larvae were found among the coarse grass seen in the foreground of the picture, but it was impossible to make any observations on the open waters. Besides this breeding-place, two others were found of quite a different character. (6) One was a pond in a field where the peat formation of the district was observed at the edges of the pond itself, and also in the brown discolouration of the water. Here larvae were obtained after breaking ice one-eighth inch thick. (c) The other was near the railway line close to the station, and was a cutting for water, having a high bank, formed by the railway track, on one side. The water was clear and flowing, but in shallow areas at the margin, where grass, duckweed and watercress provided shelter, larvae were numerous. (2) CASTLETOWN, November 30th. (a) Near the public park, and separated from it by the river (Silver Burn), is a meadow with a stream running through it; this stream at the time of our visit was overflowing at various points, covering a large area with shallow, stagnant water. Only at one end of the flooded area, in a boggy patch, with hoofprints, was a single larva obtained after considerable searching, and none was found in the stream itself, which was moving fairly rapidly. (6) In striking contrast to the absence of larvae from the centre of the meadow, was their abundance in a narrow ditch on the side remote from the river (Photo. 3). In the photograph the ditch in question is on the right, while the flooded area extends to the left for a considerable distance outside the picture. In this ditch, which varied in width from 3 feet to about a foot in some parts, and which was a foot deep with over- hanging banks, sixty larvae were collected in ten minutes by one person using a small scoop. The vegetation was chiefly grass and watercress, and from the abundance of life of various kinds present we were led to conclude that this formed a permanent and very suitable breeding-place for A. defurcatus. (3) DouGLAs, November 18th. In the neighbourhood of Douglas outside the Nunnery estate, about twenty minutes’ walk from the quay, an extensive breeding- 78 ground is present. It is that portion of flat land which lies to the right of the path outside the Nunnery towards the racecourse. The only indication of the marshy nature of this ground was the appearance of reeds, as the water was concealed by the vegetation and was not visible even at a few yards distance. The area is approximately 120 yards by 35 yards, and is continuous with a similar area not examined by us on the left of the path, where it turns towards the racecourse. In the area under consideration larvae were constantly found on testing in various directions at five-yard intervals, very frequently in hoofmarks. Beyond this field, across the hedge towards the racecourse, is what appeared at first sight to be a continuation of the same sort of half marshy ground, but the water present was red and contained a great quantity of flocculent material. No larvae were found here despite prolonged testing. (4) GLEN GARWICK ROAD, November 1oth. No larvae were found actually in this glen, although a swampy patch just in front of the hotel was regarded as promising, but a breeding-place occurs beside the Electric Tramway line from Groudle, just beyond the level crossing past Liverpool Arms Hotel to the left of the line going towards Garwick. Larvae were found here in a field which was marshy, and their distribution appeared to coincide with the occurrence of patches of duckweed. (5) GLEN GREENAUGH, December Ist. In a pond on the property of Balla Vale, just above the weir across the stream, larvae were found in small numbers. The pond contained a large quantity of dead leaves, and at one end where larvae were found grass was present; it is situated about 150 yards from the house. (6) GLEN GROUDLE, October 28th and November 16th. (a) On the occasion of the preliminary visit, a large breeding- place was found in the glen in the shape of an artificial pond which lies between the path and the river, and which is used as a fernery. The breeding-place is 80 yards long by 25 yards across at its widest part, and is situated about 500 yards from the viaduct. The pond is sub-divided into what is practically a series of small ponds by Annals Trop. Med. & Parasitol., Vol. XV ; PATE CXL PuotoGraeH No. 6 PuoroGraru No. 7 C. Tinling & Co., Lid.. Imp 79 means of banks of considerable width. Larvae were found with ease all over the area. The vegetation consists of coarse grass, reeds, waterlilies and, on the banks, ferns (Photo. 2). (4) On the second visit, by passing down the glen, we found a breeding-place at Groudle Creek in close proximity to the sea, separated from it, in fact, only by a bank of shingle and the banks of the river itself; it is seen in Photo. No. 4. From its proximity to the sea and from the presence of much seaweed and flotsam in this collection of water, we concluded that it must contain a considerable excess of sodium chloride, and the water had a brackish flavour; but analysis* showed that, in fact, it contained only 7°63 parts per 100,000, which is stated to be lower than many potable waters of the Isle of Man. This breeding-place, which was about 30 yards from the sea and 100 yards from the nearest croft, did not yield numerous larvae. (7) GLEN Laxey, November 2oth. Close to the entrance to Laxey Glen Gardens there is a boggy- looking field on the left of the path, in which larvae were found. The field is bordered by a wall (seen in Photo. 5), which separates it from the bandstand and pleasure ground. To the right of the position seen in this photograph are various swings and apparatus for the amusement of the numerous children visitors. The hotel lies behind the camera to the right at a distance of about a hundred yards. It may be noted again here that even at a short distance the presence of water could not be perceived, but could only be inferred from the character of the vegetation. (8) GLEN SULBy, December 7th. The Curragh region mentioned above in connection with Ballaugh extends towards Sulby Glen, and can be approached from this point. (a) At the edge of the Curragh larvae were found in a field surrounded by stagnant ditches. The centre of the field was swampy and larvae were found here, and, where the water was clear among the reeds, in one of the ditches bordering the field; in two * Kindly carried out for us by Mr. Fyffe of the Public Analysts Laboratory, Douglas, Isle of Man. 80 other ditches, where the water was loaded with red flocculent material, no larvae were discovered. * (6) Another breeding-place was found in Sulby itself, about 400 yards from the Glen Hotel. It was a marshy piece of ground beside the road, and was bordered on the far side by an area of higher ground where many trees had recently been cut down. The photograph showing this breeding-place of A. dzfurcatus shows also a very uncommon breeding-place of A. plumbeus, and will be found under the latter heading, Photo. 8. (9g) GLEN WILLYN, December 7th. In the terminal part of this glen, about 300 yards from the sea, there are pleasure grounds furnished with bandstand, swings, etc. These abut on the small stream, and just opposite them is a narrow wet strip of ground at the foot of a steep hill. In this wet area larvae were found in that portion adjacent to the fence which runs along the side of the stream, and separated from the pleasure ground by only the width of the stream. (10) GREEBA, December 5th. The railway from Douglas to Peel runs in a valley for a consider- able portion of the way. The land close to the railway is marshy, and has at intervals much free water lying on it. This strip of wet land was examined in the region of Greeba, and there was no difficulty in finding larvae. (11) PEEL, December 8th. Larvae were found in a small narrow drain at the foot of the hill bordering the road along the quay on the way to Peel Castle. The water was sluggish and much grass was present in many places. (12) PorT ERIN, November 26th. Three breeding-places were discovered in this locality. (a) The first was in marshy ground in the valley near the far end of the golf course. (6) The second was a slow-moving stream with high banks and much vegetation, chiefly grass and watercress situated between Port Erin and Ballafesson village. (c) The third was a small piece of marsh land covered with reeds in the centre of the village of Ballafesson. Annals Trop. Med, & Parasitl., Vol. XV JRL ANIME, PIE PHotoGrapH No. 8 Puorocrapn No. 9 C. Tinling & Co., Lid., Imp. 81 (13) PORT SODERICK, December 5th. About half a mile before Port Soderick, on the road from Douglas, beside the road a breeding-place was found. It consisted of a pool of clear water, deep in the centre with a rapid streamlet running through it. The vegetation was considerable, and consisted of a fairly thick layer of duckweed which covered the centre of the pool and reached to within a few inches of the edge in most places. The edges were covered with grass and duckweed. The larvae were present in numbers, and were found chiefly near the edges. (14) Port St. Mary, November 20th. Close to the station, in a meadow belonging to Ballacreggan Farm, just behind the gas works, a small ditch a foot wide, over- grown with grass and not conspicuous, proved to contain larvae. The water was clear and had a few patches of duckweed on it at intervals, and the surface of the water was about a foot below the level of the banks, which were undermined. (15) RAMSEY, November 22nd. An excellent breeding-place was encountered on the outskirts of the town. The extent of this area, a marshy field, is 120 by 120 yards, approximately. Very numerous larvae were found here. Situated about 15 yards away is the playground of a large school, which can be seen in Photo. No. 6. In the whole of this area larvae were found. (16) ST. JOHN’s, December Sth and oth. (a) Larvae were found in a pool in a field adjacent to the station. This pool serves partly as a cesspool for drainage from the station lavatories; it is situated 10 yards from the platform, 6 yards from the lavatories and about 30 yards from the main road. The water was slightly coloured, the margins very boggy, and the vegetation mostly rushes with some duckweed and grass. (6) On the road from St. John’s to Peel, in a field on the right side of the road about half a mile out of St. John’s, larvae were found in the margins of a narrow, rapid stream at the foot of a hill. (c) Further on the road, about a quarter of a mile beyond this ’ breeding-place, a considerable amount of marshy ground exists at 82 the side of the railway. There is an extensive breeding-place between the railway and the St. John’s-Knockaloe Road, the edge of the marshy ground lying 100 yards distant from the nearest farmstead. Anopheles maculipennis, Mg. A number of cowsheds, stables, piggeries, fowl-houses, latrines, outhouses, cellars, lofts, sheds and shelters of various kinds were examined with a view to finding the adults of this mosquito. (17) BALLAUGH, December 6th. (a) Three cowsheds were examined, of which two were small, dark, warm, flat-roofed, with numerous cobwebs present, and the third lofty, well-lighted, with gable roof and almost free from cobwebs. In one of the former, three, and in the last, one A. maculipennis 2 were captured. Total 49 (6) One stable of modern construction, with high gable roof, but rather dark ... is Fer ive ube sa) Hae (18) CASTLETOWN, November 30th. (a) One cowshed, small, dark, warm, low-roofed, situated in the centre of the town 4 eas pat 36 ov MOR? (6) One stable for two ‘horses, in centre of town, low- roofed and warm, with few cobwebs except in one corner ... 3 2 (c) One outhouse adjoining stable, and of same construc- tion, used as a potato store, cooler and draughty ... Sek nil, (2) Two lofts; one warm, dark, slate-roofed, with few cobwebs, situated over (&) and (c), nil; the other situated above (a), dark, slate roof, numerous cobwebs, 19. Total 1 9 (19) DouGLas, November 18th. At the Nunnery Mill the following were examined :— (a) Stable, a large, dark stable on ground floor, just being whitewashed, few cobwebs a hs bet beg iJ) Nggeme (6) Fowl-house, low and dark ie 3: te 62,6 pad (c) Cellars extensive, low-roofed, under mill Fieae fio Sail (d) Loft situated above (a), but not newly whitewashed, cobwebs numerous a eos Ame vil: (e) One stone-built cette “ise asa a towals run? beni Annals Trop. Med. & Parasitol., Vol. XV PEALE XT! PuoroGrarnH No. 10 PuotroGrarpu No. 11 83 At the back of the town:— _ (a) Stable, small, dark, flat-roofed, warm, numerous large cobwebs _.... sie oy ails (6) Outhouse adiownne, enter ae oie as ve) Til (c) Loft above (2) and (4), dark, warm, wood-roofed ... nil. (20) GLEN GARWICK ROAD, November toth. (a) One stable, disused 2 set ahs ac =e hide vi (6) One loft, above (a) ae aor a ae eee tal (21) GLEN GROUDLE, October 30th, November 15th and 16th. (a) One cowshed, situated 15 yards from a cottage (Photo. 7) and 100 yards from a_ breeding-place of A. bifurcatus was examined. It was gable-roofed but dark, low, warm and had many cobwebs. The door was central in the side wall, and at one end two cows were kept, the other end being a coal and wood store. There were captured here, distributed equally over both ends of the shed, but less numerous just at the door ie ise oe e Le. (6) One stable for two horses, dark, flat-roofed, moderately warm, but draughty... a se Ate eo 22n0 (c) Three latrines near stable oe ene (d) Four cellars situated near stable, wae in Robes and two under verandah (e) Three outhouses ... nil. (f) One loft over (4), e Aigunicating Cele the Efile Be an opening two feet square above the manger, and by ladder entrance also, well-lighted, clean and airy, recently renovated roof of corrugated iron, few cobwebs, draughty — nil. (g) Two sheds, corrugated iron, a few yards from stable, nil. containing piles of wood . eet oo. a Fa), Bue (A) One shelter Goalie) in sles se sae ee yy gh Dil. (2) One kennel abe sve vas xf Ae sees, Tbe: (22) GLEN SULBY, December 7th. Two cowsheds :— (a) One very small, two animals, gabled, corrugated iron roof, dark, warm, the roof moist, cobwebs very numerous ... 9 ? (4) Similar but wood-roofed, recently swept, few cobwebs _ nil. 84 (23) KIRKMICHAEL, December 7th. (a) Three cowsheds, flat-roofed wood, dark, warm, clean, to hold twenty, eight, four animals, respectively ais (6) One pigstye, usual type, dark, fairly warm, cae numerous (c) One loft bore frees cowshied, Riek gate coats dark, warm, many cobwebs (24) PEEL TO ST. JOHN’S, December 8th. (a) Two cowsheds :— (a) Large, sixteen animals, flat-roofed, warm, cob- webs over mangers and in corners only (a1) Large, ten animals, flat-roofed, dark, few ae webs, except in corners (6) Two stables : — () Flat wood roof, warm, dark, few cobwebs (ii) Similar but more cobwebs (c) One latrine outside house (d) One loft above (a) (11); high gable a3 col aad airy, containing straw and hay (25) PorT ERIN, November 26th and 20th. (a) Four cowsheds :— G) Two, low flat wood roof, dark, warm ... (i) Two, iron roof, low, warm, moist, dark (6) Three stables, dark, unventilated, cooler than cow- sheds. In one BS (c) One pigsty, low, dane wee poet many eoeane se (d@) Two fowl-houses, thatched low (e) One loft above (a) (1), dark, cool (26) Port St. Mary, November 29th. (a) Two cowsheds, large, dark, gable, slate roof recently limewashed, few cobwebs (6) One outhouse, continuation of cowebed ald, ‘draughts (c) Two sheds, one for straw, one for carts nil. nil. nil. 49 17 9 23 (8) nil. nil. nil. nil. nil. iy nil. nil. nil. nil. nil. nil. 85 (27) RAMSEY, November 22nd, 23rd and 24th. (a) Four cowsheds :— (i) One for six animals, low wood roof, dark, warm dae ate +3 Nae mise Sila wo SO (1) Three, first rather light, cool, corrugated iron roof; second and third warm, low wood roof, dark; inv thands. x2 has e Oe oR 5 ay (6) Four stables :— (i) One large, airy, high corrugated iron roof, few cobwebs, accommodation for about fifty horses we PD (ii) Two, small, wood roof, dark, with cobwebs... _ nil. (a1) One, airy, light,.cool, wood roof, cobwebs numerous... eth bd *. aoe sok sont atl (c) Six pigsties; usual type, in each of two, one female. mhotaks Atay Anopheles plumbeus, Steph. _ (29) BALLASALLA, December oth. Sycamore. A breeding-place was found just outside the churchyard of the Abbey Church, between it and the river. The tree was situated a few yards from the road. The aperture was at a height of 5 feet from the ground and 1} inches in diameter, depth of hole 6 inches; the water was yellow tinged. (30) BALLAUGH, December 6th. Sycamore. In the farmstead of Mr. Kneen a breeding-place of this mosquito was found in a tree situated about 30 yards from the dwelling-house. The aperture, situated 8 feet from the ground, was 6 inches in diameter and the depth of the cavity 12 inches. Small larvae only were found. (31) Douc3as, October 30th, November 18th, December 3rd. Seven breeding-places were found in and near Douglas. On the previous visit, October 30th, two breeding-places, (a) (5), were found in the small glen behind Falcon Cliff Hotel. October 30th :— (a) Sycamore, situated 15 yards from the garage and 27 yards from the main road. Aperture at 1} feet from the ground, diameter 86 about 6 inches, depth 9 inches. The whole trunk was hollow. Large and small larvae numerous. (6) Sycamore, 60 yards from garage and 10 and 12 yards, respectively, from two roads. Aperture 1 foot from ground, hole g inches deep; small larvae. November 18th :— (c) Sycamore in garden of Nunnery Mill Cottage, situated 12 yards from house. Aperture 12 feet from ground, measuring 2 by 14 inches, numerous larvae. (2) Lime in paddock below cottage and mill. Situated 30 yards from house and 15 yards from the mill. ; (e) Sycamore just outside garden of Mill Cottage. Aperture about 4 feet from ground, with a smaller aperture 3 feet from the ground almost level with the water. Owing to difficulties in either siphoning or scooping out the water in this cavity, resort was had to flushing out the larvae by pouring a bucket of water into the upper opening and catching the water as it emerged from the lower opening. December 3rd :— (f) Sycamore situated in garden of Marina Terrace, at the junction of two streets and distant 20 yards from an elementary school (Photo. No. 10). Aperture at 2 feet from ground 5 by 3 inches in size and cavity 6 inches deep. (g) Sycamore in same garden 5 yards from the previous one. Aperture 2 feet from ground 8 by 5 inches, depth 2 feet; small larvae numerous. (32) GLEN GROUDLE, November 19th. Spanish Chestnut. Just where the bridge on the high road crosses the Groudle river, a very large breeding-place was found. The aperture, which is at 10 feet from the ground, appears over the parapet of the bridge and is 9 inches in diameter, the cavity being full of red water to within 2 inches of the lip of the aperture. (33) LAxEy, November 2oth. Spanish Chestnut. \n Laxey Glen Gardens, on the right side of the arena in front of the bandstand, are some trees on a slightly raised terrace; in one of these, larvaé were found ; aperture at 6} feet 87 from the ground, 3 inches in diameter and facing upwards, depth of cavity 15 inches. (34) GLEN SuLBy, December 7th. Ash. Breeding-place found in this locality is of exceptional interest. In an area situated 400 yards from the Glen Hotel, where many trees had recently been cut down, larvae were found in the stump of an ash tree. The aperture is a foot in diameter, but before water is reached at 18 inches from the top of the stump the cavity has widened greatly, so that the surface of the water is very much larger than the aperture of entrance. Not only so, but also the cavity extends downwards until it is much below the level of the ground. That the breeding-place is a true rot-hole, and entirely contained in the tree stump, is shown by the facts that it was impossible to push a stick down through it at any place, and that the water was deep red in colour; it contained numerous larvae of Myiatropa florea and various chironomids (Photo. No. 8). This photograph is further of interest because the marshy ground below, indicated by arrow, is a breeding-place of A. difurcatus. (35) KIRK BRADDAN, December toth. Sycamore. Near Braddan Church, a tree was found with a cavity containing larvae. It stood on a bank behind the Hotel at a distance of 20 yards. Two apertures were present about 4 feet from the ground and about 5 inches in diameter. (36) Kirk MICHAEL, December 7th. Spanish Chestnut. The breeding-place found here was in a tree situated in a field to the right of the road leading to a farm (Mr. Mylchreest), and distant 12 yards from the cowsheds and 80 yards from the White House. The aperture was at a height of 7 feet and was remarkably small, about 1 inch. By means of the siphon, however, a great rush of red water was obtained from a large cavity, with larvae in numbers. (37) PEEL, December 8th. Spanish Chestnut. The nearest point at which A. plumbeus was found was the estate of Ballamore. The tree was situated beside the main road. The aperture was 9 feet from the ground and 8 inches in diameter, depth 6 inches. 88 (38) Ramsey, November 23rd. On the estate of Miltown, outside Ramsey, two breeding-places were found. (a) Sycamore, situated 15 yards to left side of Ramsey Road. Aperture at 10 feet from ground 6 by 7 inches, depth 2 feet, the water surface being about 1 foot from lip of aperture; numerous larvae (6) Sycamore. In close proximity to the foregoing, aperture at 8 feet from ground, 12 inches in diameter, a very large cavity full of a pulpy mass of débris, with a layer of water on the top. (39) St. JOHN’s, December 8th. At a house on the St. John-Knockaloe Road, just beyond Ballamore estate on the left of the road, two breeding-places in sycamores were found. (a) Sycamore. This was situated 20 yards from cowsheds and 30 yards from house; aperture at 4 feet from ground, 9 by 4 inches, depth g inches; trunk above hole also hollow for more than 18 inches up ; numerous larvae, over twenty in the first scoopful of water taken out. ‘ (6) Sycamore. Standing immediately beside a latrine, 15 yards from the house. There was a large cavity with two apertures, each about 5 to 6 inches in diameter (Photo. No. Q). Culex pipiens, L. Hibernating females were found in the following places :— Douglas, in 1 stable, 1 fowl-house, 1 cellar, 1 loft, 2 shelters, 1 latrine; Glen Garwick Road, 1 stable, 1 loft; Groudle Glen, 1 outhouse, 2 latrines, 2 cellars, 1 loft, 2 sheds; Port St. Mary, 1 shed; Ramsey, 3 cowsheds, 2 stables, 3 piggeries, 1 loft, 1 shed. Theobaldia annulata, Schr. Hibernating females were found :—Groudel Glen, 1 cellar, 1 loft, 1 latrine, 1 open shelter; Kirkmichael, 1 cowshed. Pupae. Douglas, Nunnery Mill Cottage, in wooden tub (see photo. 11). Ramsey, in association with larvae of A. dcfurcatus (Appendix, No. 15). MAP SHOWING DISTRIBUTION OF MOSQUITOES IN THE ISLE OF MAN A. bifurcatus A. maculipenms A. plumbeus KC. pipiens ART. annulata AAC. morsitans AC. fumipennis 89 BALLAUGH Oa KIRKMIGHAEL go Larvae. Ballaugh, under }-inch ice in peat bog and pond in field; Castletown, ditch; Glen Garwick, concrete artificial pond; St. John’s, manure pit; Port Erin, marshy field; Ramsey, see above, under pupae. Culicella morsitans, Theo. Larvae. Ballaugh, under }-inch ice in pond in field and peat bog, in association with 7. annulata and C. fumipennis larvae; Douglas, marshy field; Glen Garwick Road, marshy field; Glen Sulby, marsh. Culicella fumipennis, Steph. Larvae. Ballaugh, as above, under C. morsztans. REFERENCES Bracktock, B., and Carter. H. F. (1920). Observations on Anopheles (Coelodiazesis) plumbeus, Stephens, with special reference to its breeding-places, occurrence in the Liverpool district and possible connection with the spread of malaria. Ann. Trop. Med. and Parasit., Vol. XIII, pp. 421-452. Bracktock, B., and Carter, H. F. (1920). On the results obtained from surveys for breeding-places of tree-hole mosquitoes in Liverpool and neighbourhood. Ibid., Vol. XIV, pp. 115-126 gi NOTES ON SOME UNUSUAL BREEDING- PLACES OF STEGOMYIA FEASCIATA, Fasr., IN AUSTRALIA BY GB) HILL, FES. From the Australian Institute of Tropical Medicine, Townsville, Queensland (Received for publication 1 March, 1921) PLATE XV Although generally regarded as a domestic species, there are several records of S. fasczata breeding in rot-holes in trees and in water-retaining plants; this habit, however, appears to have escaped notice hitherto in Australia. On 20th January, numerous larvae and pupae of this species and of Ochlerotatus notoscriptus, Skuse, were collected from a tin containing about 5 inches of water and a quantity of decaying leaves, which was found in the dense scrub 600 yards distant from the nearest of several seaside dwellings on Magnetic Island. Several adult Stegomyia which were captured at the same time (1 p.m.) while attempting to bite, and others which- were bred from the larvae and pupae, were noticed to be distinctly darker and smaller than the forms found throughout the year in the Institute buildings. In order to determine the permanency of this dark form, a number of these males and females were bred from for five generations under the usual laboratory conditions, and concurrently with an equal number of generations from individuals of the lighter coloured form commonly found in dwellings. From both series males and females of each generation were secured for comparison, the experiment being terminated, in the case of the dark form, with the sixth generation, which comprised males only. An examination of the material thus obtained showed that the light form bred true, i.e., the individuals of each generation did not differ from their progenators, whilst the dark form produced, in each generation 92 after the first, a proportion of individuals of both forms, as well as intermediate forms. In both series some variation was noticed in the shape and width of the outer lyre-shaped thoracic ornamentation and in the length and width of the median thoracic stripes, but in none were the latter entirely absent, a character recorded by Taylor (1914). The island referred to above lies in Cleveland Bay, about four miles distant from Townsville, and was formerly utilised as a site for the pert Quarantine Station. On 5th May, a considerable number of mosquito larvae were siphoned from a rot-hole in a poinciana tree (Plate XV), growing in the Hospital grounds and distant about 70 yards from the nearest dwelling. In addition to Macleava tremula, Theobald, and Ochlerotatus quasirubrithorax, Theobald,* this collection produced a large number of S¢egomyzia fasciata, similar in size and coloura- tion to the Magnetic Island form. On 7th September, another batch of larvae and pupae were collected from this hole, and from them about one hundred adults were bred. In this case the males were of about the average size, but quite as dark as any I have seen, whilst the females were of the light form and unusually large. REFERENCE Taytor, F. H. (1914). Proceedings of the Linnean Society of New South Wales. Vol. XXXIX, p. 455. * Identified by Mr. F. W. Epwarps, as a variety of this species. ROT-HOLE IN POINCIANA TREE. BREEDING PLACE OF OCHLEROTATUS QUASI RUBRITHORAX, MACLEAYA TREMULA AND STEGOMYIA FASCIATA 1. & Parasitol., Vol. XV PEATE XV a r a . 1 ] Syeaat™ 1 AG 4 - s - a ; ' i = 7 ay ® oo % , : \ = — —-, : ‘ A re 4 ¥ - vel u a ‘ oe ' A ‘ - : ‘ ; ¥ . 33 MUSCA DOMESTICA, Liyy., AS A “BUSH FLY’ IN AUSTRALIA BY G, Ea HILL; F.E.S, From the Australian Institute of Tropical Medicine, Townsville, Queensland (Received for publication 1 March, 1921) In the literature dealing with the etiology of M/. domestica, I can find no record of this species in the réle of a ‘bush fly,’ z.e., a fly which lives and breeds normally beyond the range of human habitations, although I understand that Major E. E. Austen is of opinion that this species originated in the tropics and has thence spread to temperate climates, where it is only able to maintain itself by the fact that it has taken to living in houses. In two widely separated localities in North Australia, evidence has been gathered which .strongly suggests in one case, and proves conclusively in the other, that this ubiquitous species is not dependent upon human habitations and environments for its existence. ; During the period 1913-1917, specimens were frequently captured in the Darwin District (Northern Territory) on stock and carcasses at distances up to a couple of miles from dwellings, but for some time it was considered probable that these flies were bred under the usual conditions; later, however, examples were captured in company with Musca ventrosa, Wied. (M. nigrithorax, Stein.), and M. humilis, Wied. (M. vetustissima, Walk.) on the carcasses of freshly skinned buffaloes which had been shot in scrub country from three to six miles distant from the nearest habitation—a cattle station sixteen miles distant from the next nearest dwelling and about thirty miles from Darwin. During the summer months, M. domestica were very numerous at the station, especially in the kitchen and adjacent living room, but they were not seen on men or horses after leaving the homestead for the haunts of the buffaloes. In April, 1919, an officer of the Stock Department, Townsville (N. Q.), brought in for identification a large number of flies which had been captured in the vicinity of a slaughter-yard and upon 94 stock grazing in the locality, in some cases more than a mile from the nearest dwelling. This and later collections mvariably included a large proportion of M@. domesézca. During the months of April to October, flies of the same species were frequently captured upon grazing horses and cattle, and upon my face, hands and clothing in the Town Common at distances up to two and a half miles from habitations. On these occasions they were generally associated with J. lusoria, Wied.’ (M. australis, Macq.,? M. fergusoni, J. & B.3), and M. nebulo, F.4, (M. hilli, J. & B.5), the last named being less aggressive than the others. From October to about the end of March, M. humilis, Wied.°® (MM. vetustissima, Walk.) is the predominant species, and is certainly the most widely distributed Musca found in Australia, being as plentiful in the outer suburbs of Melbourne (Victoria) as it is in Central Australia, N. Territory, and the N.W. and Kimberley Divisions of W. Australia. In the bush or open grazing country near Townsville, M. domestica oviposits on fresh horse-droppings, but they will also oviposit and rear their progeny on decaying vegetable matter, as shown by the fact that upon two occasions I have bred adults from full-grown larvae taken in nests of the Black-throated Grebe (Podicipes novae-hollandiae), which had become stranded upon the margin of a swamp, and in which the eggs had not yet hatched. The same nests and, also, small accumulations of drift, z.e., leaves, horse- and cow-droppings, etc., blown up upon the margins of swamp, served as breeding-places for Stomoxys calcitrans and Sarcophaga sp. Major E. E. Austen, who kindly examined the specimens of M. domestica from bush localities in N. Territory and N.Q., determined the latter as a variety of the typical form. Evidently the distincion is a very fine one, since recently I have examined a much longer series than was available to that. worker, and I have compared them and their larvae with typical forms (from town dwellings) and their progeny, without being able to detect any variations peculiar to one series. (1), (4), (6) Identified by Professor M. Bezzr. (2) Identified by Major E. E. Austen. (3) Jounston, T. H. and Bancrort, M. J. Proc. Roy. Soc. Queensland. Vol. XXXI, No. 12. (5) Jounston, T. H. and Bancrort, M. J. Mem. Queensland Museum.~ Vol. VII, pt. 1. 1920. 95 NEW TSETSE-FLIES (GLOSSINVA) FROM THE BELGIAN CONGO BY 4 ProFessor R. NEWSTEAD, F.R.S. AND Miss ALWEN M. EVANS, M.Sc. (Received for publication 14 March, 1921) We have just received from our esteemed colleague, Dr. J. Schwetz, a small collection of tsetse-flies, captured by him in the Belgian Congo. Included in this collection are eleven examples (6 33, 5 29) of a new and hitherto undescribed species, all of them taken in the region of the River Kwango, on the frontier of the Portuguese territory. In his letter, dated 20.xii.20, Dr. Schwetz says that he had just visited this region for the first time, and that in returning down the River Kwango by boat he came to a region abounding in this species and Gl. palpalis, R.D. This new species belongs to the ‘Fusca Group’! of tsetse-flies, and is described below as Glossina schwetzi, sp.n., in honour of its discoverer, who has devoted long years of research into the bionomics of this important group of insects, and their relation to human trypanosomiasis. In a former communication in these Annals? attention was called to a variety or race of Glossina fusca, Walker, from the Belgian Congo, in which the female armature exhibits a marked deviation from the form of signum found in typical examples from the Gold Coast. The male genital armature of the Congo examples also differ in the form of the harpes from those found elsewhere. We are convinced, therefore, that we have to deal with a well-marked race of Glossina fusca: this we have given varietal rank under the name congolensis, var. n. GLOSSINA SCHWETZI, sp. nov. Hairs of the third antennal segment about one-sixth lo one-seventh the width of the segment. Wings of the female with the thickened portion of the anterior transverse vein darker in colour than the rest. Harpes of the 96 male divided into three processes, the proximal process short and spine-like. Female with signum of the uterus consisting of a single chitinous plate, the long axis transverse and widest in the distal third. Mate: Length, 10-11-9 mm. ; proboscis, 2-3-2°8 mm. ; width of head, 3°2-3°3 mm.; front at vertex, 0-6-0-7 mm. ; wing, II-12-4 mm. Fic. 1. Male armature of Glossina schwetzi, sp. n. s.¢., superior clasper; i.c., inferior clasper ; m.p., median process ; .p./., inferior median process ; v., vesica ; 4., harpes. : FEMALE: Length, 12:5 mm. ; proboscis, 3: mm.; width of head, 3°5-3°6 mm. ; front at vertex, 0-75-0°8 mm.; wing, 13-2-13°6 mm. Mate: Head with the posterior ‘surface ‘mouse grey’ (Austen), antennal cavity pearl-grey, sometimes with a pale vinous tinge; ocellar Vol. XV, p. 96. Paper by Newstead and Evans. Add to explanation of Fic. 1: ‘2: Ventral view of the armature of Glossina schwetzi. 3: Harpes of Glossina tabaniformis in profile.’ 97 spot and frontal stripe unicolorous pale brown ; antennae with the distal two-thirds of the third'segment infuscated, the rest pale buff ; outstanding hairs on third segment, short, about one-sixth to one-seventh the width of the segment. Proboscis bulb pale translucent buff-yellow, the upper lateral margins brownish or orange-brown, ventral median suture proxi- mally, dusky to orange-brown. Thorax with the usual distinctive colour and markings ; sterno-pleurae more or less infuscated ; scutellar bristles long. Abdomen: Dorsum of first and second segments light brown; the remaining segments, together with the lateral margins, dark brown (‘mummy-brown,’ Austen’), the distal angles either unicolorous with the rest of the abdomen or slightly paler. Legs: Light or dusky ochraceous- buff; leg I with the femora infuscated on the upper half of the inner surface, tips of the last two’segments dark brown or black ; leg II with the tips of the last two segments generally more strongly marked than in I ; leg III with the last two segments entirely dark brown or black, paler beneath, proximally. Wings with the thickened portion of the anterior transverse vein scarcely darker in colour than the rest. Genital armature : (fig 1) Harpes (h) with three bi-lateral processes; the proximal process short and distinctly spine-like ; second and third pairs long, slender, and only very slightly widened proximally ; median process scarcely projecting beyond the inferior claspers (ic) ; the inferior median process (mp) narrowly elongated and projecting backwards considerably beyond the inferior claspers. Superior claspers (s.c.) bluntly bifid. FEMALE: Colour and pattern similar to that of the male; but the distal segment of the abdomen usually paler, and the thickened portion of the anterior transverse vein of the wing darker in colour than the rest. Genital armature : (fig. 2) External armature possessing no distinctive characters, Signum of the uterus consisting of a single symmetrical chitinous plate of the form shown in the figure, the greatest width being 0-38 mm. Laterally, it bears a pair of black curved thickenings (a.th), and sometimes a second pair (f.th) lies behind the anterior pair. That portion of the plate posterior to the level of the thickenings is much more heavily chiti- nised than the anterior portion, in which a large transparent space occurs medianly. The upturned anterior processes (a.p) which are a marked feature of the signum tend to slight variation, and are much smaller in one individual than those shown in the illustration. Belgian Congo: River Kwango, Kasongo Lunda, 24.x.1920, 2 99; River Kwango, near Cuango, 24.x.1920, 4 99 (‘ Le soir par terre ’) 25.x.1920 98 3 22; River Kwango, near Kundi, 1.xi.1g20, 2 33 (‘ Le soir sur chemin ’) Dr. J. Schwetz. ‘ In its general external facies the male of this species bears a striking resemblance to that of Gl. tabaniformis, West ; the female, on the other Fic. 2. Glossina schwetzi, sp.n. 9. Signum x c.155. a.p., anterior process; a.th., anterior thickening ; p.tb., posterior thickening. (Genital fossae not shown.) hand, owing to the darker colour of the thickened portion of the anterior transverse vein, might easily pass as a specimen of Gl. brevipalpis, Newst. Both sexes may, however, be readily distinguished from any other members of the “ Fusca Group’ by the strikingly characteristic genital armature. Cc of Pics: Glossina schwetzi, sp. n., 4., antennal fringe x c. 325; B., third segment of antenna x c. 40. _ Glossina tabaniformis, C., antennal fringe x c. 325}; D., third segment of antenna X c. 40. Furthermore, the antenna of Gl. schwetzi can be distinguished from that of Gl. tabaniformis by the shorter fringe of fine hairs on the anterior edge of the third segment (fig. 3). | The ratio of the length of the longest fringe-hairs to the greatest width of the segment was determined oF in four specimens of each of the above-named species. They are tabulated below : ; GI. schwetzi, sp. n. ... es Nan 6) 1:62 berg To 75 Gl. tabaniformis ... cos VBE Ry Tdok Es As3 1:43 COMPARATIVE TABLE OF THE MORPHOLOGICAL CHARACTERS OF THE MALE GENITAL ARMATURE GI. schwetzt Gl. tabaniformis Harpes : Fig. 1 Fig. 1, no. 3 Proximal process ... A stout spine, one-fourth Slender and equal in length of second. length to the second. Second process... ... Slender. The same. Third process (distal) ... Long, slender and simple. Long and strongly bifur- cate. COMPARATIVE TABLE OF THE MORPHOLOGICAL CHARACTERS OF THE FEMALE GENITAL ARMATURE Gl. schwetzi Gl. tabaniformis Signum of uterus... Boe Fig. 2 With a bilateral pair of Lyriform. No bilateral black curved thicken- curved thickenings. ings. Long axis... oe ... Transverse. Longitudinal. GLOSSINA FUSCA var. CONGOLENSIS, var. nov. Colour and pattern generally as in typical Gl. fusca, Walker, but the wings are usually more heavily infuscated. — Harpes of male (fig. 4) with the proximal process reaching to the tip of the second (serrated) process. Bilateral portions of the signum of the uterus of the female sub-rotund. MALE : Genital armature (fig. 4). | Harpes (A) with the narrow proximal process reaching to the tip of the second, serrated process; the latter relatively narrow, and more or less suddenly truncate distally ; the third, a somewhat sickle-shaped appendage, with the distal arm very much shorter than the proximal one. FEMALE: Genital armature. In the signum of the uterus (fig. 5) the width is greater than the length; the bilaterally symmetrical plates forming the main portion of the signum sub-rotund, and separated medially by only a very shallow depression. 100 The foregoing description, and also the illustrations which accompany it are based upon fourteen examples, representing both sexes, captured in Fic. 4. (1) and (2) Male armature of Glossina fusca var. congolensis, vat. n. (3) Harpes of Glossina fusca. sc. superior clasper ; 7.¢., inferior clasper ; ., vesica; 4., harpes ; i.o., intromittent organ ; h.1., sickle-shaped process of harpes. the Katompe, and in the Lomami-Kisengwa districts of the Belgian Congo by Dr. J. Schwetz. IOI Fic. 5. Glossina fusca var. congolensis, vac.n. @. Signum X c.155. ¢.p., anterior plate ; cr., crescentic thickening ; f., petaloid area. (Genital fossae not shown.) pea ae See me ee Fic. 6. Glossina fusca, Walker. 9. Signum x c. 155. cr., crescentic sclerite ; ¢., half of main portion of signum; g./., genital fossae ; p., posterior plate. 102 COMPARATIVE TABLE OF THE MORPHOLOGICAL CHARACTERS OF THE MALE GENITAL ARMATURE G. fusca Fig. 4, no. 3 ... Reaching to the middle distance of the serrated process. Harpes : Proximal process Serrated process ... Distally broadly lanceo- late; shaft relatively broad. ... Distal arm of sickle- shaped process as long as the proximal one. Distal process ... G, fusca var. congolensis Fig. 4, no. 1, 2 Reaching to the tip of the serrated process. Distally suddenly trun- cate; shaft relatively narrow. Distal arm less than half the length of the proximal one. COMPARATIVE TABLE OF THE MORPHOLOGICAL CHARACTERS OF THE FEMALE GENITAL ARMATURE Gl. fusca Signum Fig. 6 Halves of main portion ... Sub-conical. Very mark- edly convex, sepa- rated anteriorly by deep V-shaped depres- sion extending to nearly one-third of their depth. Chitinisation ... Generally complete, the colour varying from deep ochraceous to black. Anterior plate (c.p.) . Absent. Greatest width . Average 0°56. Extremes 0°60, 0°46. REFERENCES 1.—Newsreap, R. (1911). Bull. Ent. Research, Vol. 11, pp. 9-36. 2.—Evans, ALWEN M. (1919). Gl. fusca var. congolensis Fig. 5 Sub-rotund. Slightly convex, separated an- teriorly by very shal- low depression. Generally incomplete, ochraceous _ petaloid area (p.) often sur- rounded by consider- able almost colourless area. Generally present. Average 0:42. Extremes 0°44, 0°40 Ann. Trop. Med. Parasit., Vol. XIII, pp. 31-56. 3-—Ausren, Ernest Epwarp (1911). A handbook of the Tsetse-Flies (genus Glossina). 103 One A; COLLECTION | OF, PAPPATACI FLIES (PHLEBOTOMUS) FROM INDIA BY PrRoFEsSOR R. NEWSTEAD, F.R.S., AND Major J. A. SINTON, V.C., I.M.S. (Received for publication 14 March, 1921) The collection of insects which form the subject of this communication was made by one of us (J. A. Sinton) in the North-west Province of India, with the exception of the single capture of a specimen of Phlebotomus major, Annandale, at Simla. It embraces the records of a very large number of individuals (664) representing three species and one variety, and, as the pappataci flies of the region in question have not: hitherto been investigated, it seemed desirable that a complete list of the captures should be recorded, the subject being a cognate one to that of pappataci fever. The occurrence of Phlebotomus sergenti, Parrot, is of much interest, the species being a new addition to the fauna of India. ‘In the examination of the long series of examples of Phlebotomus minutus, Rond., it was found to be impossible to draw any hard and fast lines for the separation of this species from P. antennatus, Newst.,' since we found so many intermediate forms ranging between typical examples of each. It may be recalled that the erection of P. antennatus was based chiefly upon the remarkably short antennal segments, and the relatively shorter legs; the male genital armature on the other hand being absolutely identical with that of typical P. minutus. It seems, therefore, that the differences in the antennal segments can no longer be considered as of specific importance ; P. antennatus has therefore been placed as a variety of P. minutus. We tender out thanks to Miss Alwen M. Evans, M.Sc., for her assistance in the preparation and determination of the material. 104 Phlebotomus papatasti, (Scop). In the males twelve examples were found to possess an additional spine on the inferior clasper of the genetalia; in some instances the spine was bilateral, in other cases unilateral. One example also possessed a supernumerary spine on the left superior clasper. ; INDIA: NORTH-WEST FRONTIER PROVINCE. DERA ISMAIL KHAN, 2 33, 6 29, bedroom, bungalow, September and October, 1919; 6 33, 7 92, ditto, 14.iv.1920 to 14.v.1920; 40 33, 52 99, cowshed, 8.111.1920 to 31.iii.1920 ; 35 3d, 74 29, ditto, I.iv.1920 to I4.iv.1920; 3 dd, 10 29, ditto, 15.iv.1920 to 15.v.1920 ; 14, tents near I.M.S. bungalow, 16.iii.1920 ; 4 3d, 11 29, ditto, 12.iv.1920 to 15.v. 1920. TANK, I 6, 5 29, inside tents with floors sunk 3 feet, 29.viii.19Ig, I.ix.1919, 16.ix.19g19; 2 29, ditto, 6.iv.1920. BANNU, 4 3d, I 9, bedroom, bungalow, I1.vili.19g19 to I2.viii.t9tg. IDAK, TocHI VALLEY, I 9, mud barracks, 26.vili.1919. Saccu, near DERA IsmMAIL KHAN, 19, stable, 14.iii.1g20. JATTA Post, I 9, mud barracks, 7.iv.1920. Murtaza, I dg, 4 99, mud barracks, I.ix.I9gIg ; I d, 6 99, ditto, 7.iv.1920. HATHALA, I dg, I 9, mud barracks, 29.Vill.I9Ig ; I Q, ditto, 6.1v.1920. Phiebotomus minutus, Rondani. The specimens showed a marked variation in colour; every form intervening between the pale typical examples and distinctly dark or melanic forms were observed. The left superior clasper in one male possessed’ a supernumerary spine. The wings of this species also showed a marked tendency to variation, both as regards size and contour, and also in the relative length of the anterior forked vein. INDIA: NORTH-WEST FRONTIER PROVINCE. DERA IsMAIL KHAN, 118 36, 34 29, bedroom, bungalow, September and October, 1919 ; 6 33, 2 29, ditto, 14.iv.1920 to 14.v.1920 ; 3 3d, 7 99, cowshed, 8.111.1920 to 31.iii.1920; 15 dd, 8 99, ditto, I.iv.1920 to I4.1v.1920; I 6, 3 929, tents near I.M.S. bungalow, 16.iili.19g20 ; 36 33, 13 9, ditto, I2.iv.1920 to 15.v.1920. TANK, 3 33, 6 99, tents with floors sunk 3 feet, 29.vlll.19I19g, I.ix.I919, 16.ix.1919. BANNU, 37 36, 37 99, bedroom, bungalow, II.viii.Ig1g9 to 12.viii.1g19. IDAK, Tocut VALLEY, 2 3d, mud barracks, 26.viii.1919. KuHIRGI, 2 99, tents, 30.viii.tgtg. HATHALA, 3 99, mud barracks, 29.viil.1919. 105 Phlebotomus minutus var. antennatus, Newst. Phlebotomus antennatus, Newst. Bull. Ent. Res., Vol. ILI, p. 365. The variation in colour from light to dark or melanic forms was similar to that found in P. minutus, but none possessed the intense lapis-lazuli colour found in the West African forms.° InpIA: NORTH-WEST FRONTIER PROVINCE. DERA ISMAIL KHAN, II 33,3 22, bedroom, bungalow, September and October, 1919; 2 34, ditto, 14.iv.1920 to 14.v.1920 ; 2 3d, 1 9, cowshed, 8.11i.1920 to 31.11.1920 ; I 3, 2 92, ditto, L.iv.1g20 to 14.iv.1920; I 3, 2 99, tents near I.M.S. bungalow, 1I2.iv.1920 to I5.v.1920, TANK, 3 99, tents with floors sunk 3 feet, 29.vili.1919, I.ix.I9Ig, 16.1x.1919. BANNU, 2 33, 7 29, bedroom, bungalow, IlI.viii, 1919, I2.vili. 1919. Phlebotomus sergenti, Parrot. Parrot? first described this species from Algeria ; subsequently it was found by Marzinovsky? at Tiflis, by Sinton in Persia, and by Buxton‘ in Mesopotamia. Hitherto it does not seem to have been recorded from India. INDIA: NORTH-WEST FRONTIER PROVINCE. DERA IsMAIL KHAN, I 3, cowshed, 8.iii.1920 to 31.il1.1920; I 3, tents near I.M.S. bungalow, 12.1v.1920 tO 15.v.1920; 3 dd, I 9, stable, TANK Roap, 7 miles from Dera IsMaIL KHAN, 6.iv.1920. Mur?TAzA, I 3, mud barracks, I.ix.1gIQ. Phiebotomus major, Annandale. Records Indian Mus., Vol. V, p. 46 (I9I0). INDIA: SIMLA, I 9, 30.v.1g20. Colour and general facies resembling those a the female in P. papatasii, but the wings are relatively longer and narrower. Position of abdominal hairs doubtful owing to the rubbed condition of the dorsal surface, but the hairs on the venter more or less erect. Palpi with the 2nd, 3rd, and 4th segments sub-equal in length ; the 5th twice the length of the 3rd. Total length of head and body, 3:5 mm. ; length of wing, 3:3; greatest width, 1-05 mm. ; length of leg ii, 4-8 mm.; femur more than half the length of the tibia, the latter equal in length to the first three proximal segments of the tarsus. This, as may be gathered from the above details, is an exceptionally large species, and in its general facies and certain structural characters agrees with Annandale’s description of his P. major (I.c.). There are some 106 important discrepancies however : i.e., in the arrangement of the abdomi- nal hairs, and the length of the body as compared with that of the legs. We feel, however, that these may be considered as possible mutations within the range of variation of the species, and have therefore placed it here. REFERENCES 1.—NeEwstTeaD (1912). Bull. Ent. Res., Vol. III, p. 365. 2.—Parrot (1917). Bull. Soc. Path. Exot., X, p. 564. 3-—Marzinovsky (1917). Medical Review, Moscow, LXXXVII, p. 612. 4.—Newsteap (1920). Bull. Ent. Res., Vol. XI, p. 307. 5.-———— (1920). Bull. Ent. Res., Vol. XI, p. 305. _ 107 NOTES ON ORIENTAL SORE IN RUSSIAN TURKESTAN AND THE RESULTS OF TREATMENT WITH INJECTIONS OF TARTAR EMETIC SOLUTION BY J. A. SINTON, M.D., Major, I.M.S. (Received for publication 27 April, 1921) The cases reported below seemed worthy of record on account of the rapidity with which they reacted to treatment with intravenous injections of solutions of tartar emetic, a treatment which does not seem to have been tried previously in Turkestan. This disease is very widely spread in Russian Turkestan, and is said to be found in all the towns on the Trans-Caspian Railway from Askhabad to Tashkent. Yakimoff and Schockov (1915) described it at Askhabad, Boukhara, Samarkand and Terméze, and in the latter place it seemed especially prevalent. Locally it is said that practically every person residing in Turkestan for five years has the marks of at least one of these sores. At Tedjen station, on casual inspection, about 50 per cent. of the people seemed to have marks of old sores on their faces, but the authors quoted above state that only 58°2 per cent. of the sores examined by them were leishmaniasis. The same authors report two cases of cutaneous leishmaniasis in dogs in Turkestan. In all the six cases noted below Leishmania tropica were found. The treatment used was intravenous injections of 2 per cent. solution of antimonium tartaratum (tartar emetic) in normal saline solution; the sores received no special local treatment. Except for a little vomiting immediately after the injection, in two cases, no constitutional effects were caused by the injections. 108 _ NOTES ON THE CASES Case 1. Russian Nursing Sister at Kaakha, Turkestan. History. Resident in Turkestan for some years. Duration of sores about three months. Condition. (a) A small raised nodular sore }-inch in diameter at the angle of the left jaw, with slight ulceration in the centre. (>) Three similar sores on the left forearm. Treatment. 5.10.18. Intravenous injection of 3 c.c. T.E. solution. 7.10.18. Injection of 4 c.c. T.E. solution. Result. On account of military operations the case was not seen again until 14.11.18, at which time all the sores were healed and the patient reported that they had all healed within three weeks after the injections. Casz 2. Sepoy A.S. History. Patient has been in Turkestan about three months. Duration of lesion uncertain. Condition. Sore about $-inch diameter on right hand. Warty and ulcerated. Treatment. 19.:1.19. Intravenous injection of 2 c.c. of T.E. solution. 22.1.19. Injection of 3 c.c. of T.E. solution. Result. Completely healed by 27.1.19. Case 3. Sepoy M.K. History. Had a small abrasion on the back of the left hand about 1.10.18, at Meshed, N. Persia, which gradually got bigger. He arrived in Turkestan on 10.10.18. Condition. On 7.1.19, at Bairam Ali, Turkestan. A large sore about 1} inches in diameter on back of left hand with inflamed margins and a warty base. No enlarged glands or thickening of the lymphatic vessels. Treatment. 14.1.19. Injection of 2 c.c. of T.E. solution. 16.1.19. E 4 ees GAS loi - 19.1.19. ss “ihgspatciorg) & 3 22.1.19. a 9» 5 c.C. ” » Result. The warty growth gradually shrivelled and the ulceration disappeared. Completely healed on 28.1.19. Case 4. Sepoy S.S. History. Got an abrasion of the left hand at Dushak, Turkestan, on 14.10.18, which gradually increased in size. Duration three months. Condition. (a) A large circular lesion about 2} inches in diameter on back of left hand with infiltrated edges and a warty base with slight ulceration in the centre. (4) A few small non-ulcerated nodules in the adjacent skin. Treatment. 14.1.19. Injection of 2 c.c. of T.E. solution. 16.1.19. Ek dla eho ehh uf 19.1.19. sib Lg iqncie: Ure; = 21.1.19. sot uae SECO. kage es Result. After the second injection, a firm nodule the size of a pea was discovered in a lymphatic vessel about 24 inches below the elbow at ‘the back of 109 the left forearm, and a soft swelling about 2 inches by 1 inch on the inner side of the same forearm. This swelling was on the opposite side to that on which the injections had been given. Marked improvement was noted at the time of the third injection, the sore being dry in the centre and the warty growth shrivelling. The sore was completely healed on 29.1.19. Case 5. Sepoy C.D. History. Said to be of two weeks’ duration. No history of wound or abrasion. Patient has been several months in Turkestan. Condition. At the beginning of January, 1919. (a) A circular lesion with thickened edges and a warty base about 1 inch in diameter on front of left wrist. (6) Two small nodules in the lymphatic vessels about two inches above the sore. Treatment. 14.1.19. Injection of 2 c.c. of T.E. solution. 16.1.19. i a BCLs | 55 4 19.1.19. les LE igs = 22.1.19. ime asStG:Cuiss A Result. The warty growth rapidly shrivelled and the nodules disappeared from the lymphatics. Completely healed on 27.1.19. Caszt 6. Sepoy N.S. History. He has been in Turkestan four months. He states that the lesions began with great itching and appeared more or less simultaneously, Duration about one month. Condition. On 1.1.19. (a) On the front of the left wrist was an oval sore about 1 inch by 3 inch, with thickened edges and a warty centre, but no ulceration. (6) On the front of the right forearm : (1) A circular lesion, 1 inch in diameter, with heaped-up and thickened edges, having an ulcer }-inch in diameter in the middle with a granulating base. (2) A small sore }-inch in diameter, with a thickened raised edge and a minute ulcer in the centre like a boil. (3) There were two small nodules in the lymphatic vessels about two inches below these sores, and there were no enlarged glands. Treatment. 16.1.19. Injection of 1 c.c. of T.E. solution. 19.1.19. ” » 3 CC. »” ” 22.1.19. »” 9» 3CC. 55 ” Result. At the time of the first injection some of the sclution passed under the skin because the patient suddenly moved his arm. At tl e time of the second injection there was marked thickening and some inflammation at the site of the previous injection. Improvement was noticed in the sores on 22,1.19, and this was marked on 25.1.19. There was still some infiltration of the area around the point of first injection on 25.1.19, but there was no suppuration. Completely healed on 30.1.19. T1o These cases may be summarised as follows :— Taste I. | | Duration Amount Number Duration of Number of Case before of T.E. of treatment days till No. treatment —_| injected injections in days cured. —— cgms. I 3 months 14 2 3 ? 21 2 ? Io 2 4 $ 3 34 months 28 4 9 14 4 3 months 28 4 9 15 5 + month 28 4 9 13 | 6 I month | 14 3 im, 14 Average 2:2 months | 20°33 | 31 68 1471 An analysis of the clinical signs in these six cases shows that Cases 2, 3, 4, 5 and 6 correspond to the ‘cutaneous hypertrophic non-ulcerating papillomatous’ type of da Matta’s classification (1916) of the leishmaniases, but in this type in these Turkestan cases it was found thai after a certain time these warty growths tend to ulcerate in the centre. As they bleed very easily, this ulceration seems most probably due to injury followed by a secondary infection. In Cases 1 and 6 the typical ‘Oriental Sore’ of the text-book was found. In Cases 1, 4 and 6 the lesions were multiple, and in Case 6 both the papillomatous type and the typical oriental sore were found. Yakimoff and Schockov (1915) state that a single sore is commonest, but as many as seventeen may occur. In Cases 4, 5 and 6 nodules were present in the lymphatic vessels within a few inches of the sores, those in Case 6 being remarkable in that they were distal to the lesions. The attached table gives a comparison between the results of treatment with intravenous injections of tartar emetic solution only, in the cases reported by myself (1917), in those reported by Greig (1917) in the same year, and in the present cases. IIt Taste II. Sinton, 1917 | Greig, 1917 Sinton, 1919 | Where contracted... ... ...| N.W. Frontier, Mesopotamia Russian Turkestan India and Mesopotamia Number of cases treated ... “a 6 | 18 6 Pn ee Average duration of the disease before treatment Rast ry tess 2 months 26 months 2:2 months Duration of the treatment in days :-— Average... see “ee 20 68 MAXIMUM sss sca ves 37 : 9 Minimum ... ace 4 8 ose 3 Number of days from commence- ment of treatment till cured :— . Average... usd A 28-3 28-2" IfT MaximUmi ss, een cos sr car 21 (2) Minimum ... ... x 12 16* 8 Amount of T.E. given intravenously :— AMETARED: tee. Tutt Pas 38-2 cgms. 71-5 cgms. 20°33 cgms. Maximum ... ou ses 89 cgms. 150 cgms. 28 cgms. Minimum ... ae a5 12 cgms. 20 cgms. Io cgms, Number of injections given :— Average... ery ae 53 ? 3 Maximum ... Se =A 9 ? 4 Minimum s..7 nat q) 2 2 Result :-— Cured... Ar yan a 6 17 6 Not cured axe ae oan ° I ° *Note.—These figures are probably excessive, as they represent the ‘ number of days under observation in hospital.’ 112 From this table it will be seen that the amount of tartar emetic needed to produce a cure in the Turkestan cases was only half to one-third of that needed in the other cases, and the rate of cure was almost twice as rapid. Although it is not possible to draw any definite conclusions from so few cases, yet it would seem probable that the variety of sore found in Turkestan is more amenable to treatment with antimony than the Mesopotamian type. I am indebted to Dr. Minkavitch, of the Russian Medical Service, for the following details of the treatment adopted for the cure of this disease by some medical practitioners in Turkestan. This treatment is a modification of the local native treatment. A piece of Emplastrum Cantharidis is cut slightly larger than the sore; this is placed over the sore and completely covered by a larger piece of adhesive plaster to hold it in position. This dressing is renewed daily, and the blisters which have formed are opened. This treatment is continued for four days, after which the sore is treated with some simple ointment. Some very good results are said to be obtained by this treatment, but the scar left is large. REFERENCES Da Marta, A. (1916). Tableau synoptique de la classification des Leishmanioses. Bull. Soc. Path. Exot., Vol. IX, pp. 101-102, quoted in Trop. Dis. Bull., Vol. IX, p. 231. Greic, E. D. W. (1917). Summary of the results of the observations on the treatment of Oriental Sore by Antimonium Tartaratum. Ind. Fourn. Med. Res., Vol. V, pp. 394-400. Sinton, J. A. (1917). The treatment of cutaneous Leishmaniasis with intravenous injections of tartar emetic. Jud. Med. Gaz., Vol. LII, pp. 239-241. Yaximorr, W. L., and Scuocxov, N. F. (1915). | Leishmaniose cutanée (bouton d’orient) au Turkestan Russe. C. R. Soc. Biol., Vol. LXXVIII, pp. 107-109, quoted in Trop. Dis. Bull., Vol. VI, p. 225. 113 A. METHOD FOR THE CULTIVATION OF BLASTOCYSTIS BY HARVEY P. BARRET From the Laboratory of The Charlotte Sanatorium, Charlotte, N.C. (Receved for publication 13 April, 1921) Quite a good deal of uncertainty has existed as to the true nature of blastocystis. Prowazek (1904) and Ucke (1907) first described this organism as a cyst of trichomonas. Alexeieff (1911) was the first to describe blastocystis as a distinct organism of a vegetable nature. Others have described this organism and discussed the possibility of its being a cyst of trichomonas or other intestinal parasite, or some degeneration form of these parasites. Bohne and Prowazek (1908), Benson (1909), Wenyon (1910), Prowazek (1911), Brumpt (1912), Chatton and Lalung Bonaire (1912), Macfie (1915), Swellengrebel (1917), Chatton (1917), have all given various descriptions of this organism. Wenyon (1920) gives a very comprehensive review and digest of the Sage Parts of his article are quoted below :— _ ‘What then is the blastocystis which occurs so commonly in the human intestine and that of other animals? Prowazek (1911) maintained that they were cysts of trichomonas, but there is no evidence to support this view. Swellengrebel (1917) has suggested that they are degenerate forms of various intestinal protozoa, while .Jepps and Dobell (1918) have noted that certain degenerate forms of Dientamoeba fragilis resemble dead blastocystis. I myself have, for want of evidence to the contrary, always regarded them as of a vegetable nature, and this may be the case in spite of the resemblance to the cysts of Prowazekella lacertae. Swellengrebel’s conclusion is that blastocystis “ peculiar form of degeneration to which representatives of different genera of intestinal protozoa may be lable.”’ Wenyon concludes, ‘It is evident, therefore, that there is a difference of opinion is not the name of a zoological genus but of a as to the true nature of blastocystis, and we must await further information.’ 114 The writer has had under observation for the past few years a case of baJantidial infection, whose stools are loaded with blasto- cystis as well as balantidia. While attempting to cultivate the balantidia, the blastocystis was found to multiply readily on the medium used. Since then numerous cultures from different individuals have been made. This work was begun in 1919, and the first successful culture made in September of that year. The universal occurrence of this organism may be noted from the articles by Lynch (1917) in the United States, Wenyon and O’Connor (1917) in Egypt, Haughwout (1918) in the Philippines, and Maplestone (1921) in Australia. METHOD The culture medium used in this work is very simple, being made up of human blocd serum and 0’5 per cent. of sodium chloride solution. Various concentrations of the serum in salt solution have been tried. The most favourable strength has been Io per cent. Walker (1913) found 0’5 per cent. salt solution to be the proper tonicity for Balantidzum coli, and this concentration has been used in the work with blastocystis. The salt solution is sterilized in the autoclav and the serum added after inactivation at 55° C. for one half-hour. The pooled serum of several individuals has been used instead of that from a single individual, although no work has been done to show whether or not one serum may be inhibitory while another is favourable to the growth of blastocystis. The sera of animals other than man have not been tried. This medium is faintly alkaline to litmus. The medium 1s distributed in narrow _ test tubes in quantity sufficient to give a column of fluid at least 100 mm. high. No growth takes place at the surface of the tube, and the parasites multiply best at the lower portion of the tube, evidently needing little free oxygen for their growth. In making the initial inoculation, a small portion of stool or an emulsion of faeces in salt solution is placed at the bottom of the tube containing the culture medium. The culture is incubated at 37° C. for twenty-four to forty-eight hours and then examined for blastocystis. It 1s best to examine early cultures every twenty-four 115 hours in order to note the degree of growth of blastocystis and to make transfers before bacterial contamination is too heavy. If good growth is obtained after twenty-four hours, a transplant is then made into fresh culture medium, using the sediment in the original tube. When the organisms have established themselves in the new medium, it is best to make transfers every forty-eight hours. Cultures seventy-two hours old, or older, will hardly ever give good growth in succeeding transfers. As stated above, the optimum growth takes place in the lower portion of the tube. In making all subsequent transfers the sediment from the cultures is used, and is introduced into the bottom of the tubes of the fresh medium. Using this method, the writer has carried blastocystis through more than twenty-five sub-cultures, covering a period of about fifty days. Cultures have been abandoned, usually because of pressure of other work or because of accident or carelessness in making transfers, although they often die from bacterial overgrowth. With proper care a culture can probably be carried on indefinitely. The writer has made no attempt to classify different strains of blastocystis though the gross appearance of organisms from different individuals would suggest strongly that several strains of blastocystis have been encountered. Nor has any attempt been made to trace out a developmental cycle for blastocystis. This task is properly left to one proficient in systematic mycology. In cultures, budding, as described by Alexeieff, and binary division are the methods of multiplication exhibited by this organism ; both of these processes have been observed in all cultures. In the examination of hundreds of preparations the so-called multiple division form of Alexeieff, pictured by Wenyon and O’Connor (1917), has been encountered on only two occasions. This form would seem to be some freak in the development of the organism rather than a true developmental phase. In cultures, as in the intestinal contents, a great variation in the size of the organisms is observed. It may be said with a reasonable degree of certainty that the large vacuolated forms are the result of degeneration, as these forms are commonly found in old cultures, and successful transfers cannot be made from cultures made up of these forms. No flagellate forms, and no forms showing amoeboid or other motion have been encountered. Blasto- cystis retains its original form after twenty-five successive transfers . 116 in artificial culture medium, and no forms were seen in the twenty- fifth transfer that were not seen in the first. _ This werk is submitted as proof that blastocystis is a distinct zoological genus, and not a cyst of protozoa nor a form assumed by protozoa undergoing degeneration. REFERENCES Avexeierr, A. (1911). Sur les ‘ Kystes de Trichomonas intestinalis’ dans l’intestin des Batraciens. Bull. Sci. France et Belgique, Vol. XLIV, p. 333- (1911). Sur la nature des formations dites ‘ Kystes de Trichomonas intestinalis.’ C. R. Soc. Biol., Vol. LXXT, p. 296. Benson (1910). Untersuchungen tiber Trichomonas intestinalis und vaginalis des Menschen. Arch. f. Protistenk., Vol. XVIII, p. 116. Boune unp Prowazek, S. (1908). Zur Frage der Flagellatendysenterie. Arch. f. Protistenk., Vol. XII, p. 1. _ Brumpt, E. (1912). Colite a Tetramitus mesnili (Wenyon 1910) et colite a Trichomonas intestinalis Leuckart 1879. Blastocystis hominis, n. sp. et formes voisines. Bull. Soc. Path. Exot., Vol. V, p. 725- Cuatron, E. (1917). Les ‘ Blastocystis’. stades du cycle évolutif de flagelles intestinaux. C. R. Soc. Biol., Vol. UXXX, p. 555. Cuatton, E. er Latunc-Bonnarre (1912). Amibe limax (Vahblkamfia n. gen.) dans V’intestin humain. Son importance pour l’interprétation des amibes de culture. Bull. Soc. Path. Exot., Vol. V, p. 135- Haveuwotr, F. G. (1918). The tissue invasive powers of the flagellated and ciliated protozoa, with especial reference to Trichomonas intestinalis. A critical review. Philippine Fourn. of Science, Sec. B, Vol. XIII, p. 217. ‘Jeres, M. W., and Dozext, C. (1918). Dientamoeba fragilis n.g., n. sp. a new intestinal amoeba from man. Parasitology, Vol. X, p. 352. Lyncu, K. M. (1917). Blastocystis hominis ; its characteristics and its prevalence in intestinal content and faeces in South Carolina. ‘Fournal of Bacteriology, Vol. II, p. 369. Macriz, J. W. S. (1915). A case of dysentery in a monkey in which amoebae and spirochaetes were found. Ann. Trop. Med. and Parasit., Vol. IX, p. 507. Maptestone, P. A. (1921). Human Intestinal Protozoa in North Queensland. Ann. Trop. Med. and Parasit., Vol. XIV, p. 283. Prowazek, S. (tg04). Untersuchungen iiber einige parasitische Flagellaten. rb. a. d. Kais. Gesundbeitsamte, Vol. XXI, p. 1. —— (1911). Zur Kenntnis der Flagellaten des Darmtraktus. Arch. fur Protistenk., Vol. XXIII, p. 96. ~ SweLteNGREBEL, N. H. (1917). Observations on Blastocystis hominis. Parasitology, Vol. IX, , Pp: 451. Waker, E. L. (1913). Quantitative determination of the balantidicidal activity of certain drugs and chemicals as a basis for treatment of infections with Balantidium coli. Philippine Fourn. of Science, Sec. B, Vol. VIII, p. 1. Wenyon, C. M. (1910). A new flagellate, ‘ Macrostoma mesnili’ n. sp., from the human intestine, with some remarks on the supposed cysts of ‘ Trichomonas.’ Parasitology, Vol. III, p. 210. (1920). Observations on the intestinal protozoa of three Egyptian lizards, with a note on a cell-invading fungus. Parasitology, Vol. XII, p. 350. Wenyon, C. M., and O’Connor, F. W. (1917). Human intestinal protozoa in the Near East. Wellcome Bureau of Scientific Research, London. 117 THE INCIDENCE OF INTESTINAL PARA- SITES, ESPECIALLY WITH REGARD TO THE PROTOZOA, AMONGST SYMPTOM- LESS CARRIERS IN JAMAICA. BY HENRY HAROLD SCOTT, Miao Mon... Logd.), © .o.E.,, PE. GOVERNMENT BACTERIOLOGIST, HONG KONG; LATE GOVERNMENT BACTERIOLOGIST, JAMAICA, B.W.I. (Received for publication 13 April, 1921) The objects of this investigation were, firstly, to find out the incidence of infection with various species of intestinal parasites in patients admitted to hospital for conditions totally unconnected with dysentery or other intestinal complaints, z.e., symptomless carriers, and, secondly, to see what strains of amoebae were common. That three successive examinations should indicate freedom from infection was a purely arbitrary standard, and is shown later to be erroneous in some at least of the cases in this series. We must bear in mind that a ‘negative examination’ does not mean that no cysts are present; it, of course, merely implies that none have been found, and, when we consider how small a proportion of the whole stool is submitted to examination, we can understand that 1f the cysts are few in number they may easily be missed in such a limited examination. Again, if only one or two are found after thorough examination of several specimens, though the results will be erroneous from the statistical point of view if put down as negative, owing to none being found in the first three specimens, nevertheless the infection is probably so slight that for all practical purposes the stool is negative. In this connection, however, it is also important to remember that the value of a negative report on examination will differ somewhat according to the species of parasite. Thus, a negative examination in respect of Giardia intestinalis is more likely to be correct, as 118 regards inference, than one in respect of Entamoeba coli, owing to the more general and even distribution of the former in a stool; while the latter are often so irregularly distributed through the faeces that one sample taken for examination is by no means representative of the whole stool. Under conditions such as the latter, two observers examining different portions of the same stool might give diametrically opposite reports: the one that he could not find any, the other that coli cysts were abundant. In cases of Giardia infection this is less likely to occur. In order to minimise this difficulty as far as was practicable, instead of taking these specimens from a stool and examining direct, { have adopted the concentration method of Cropper and Row (1917). By this means, not only was the examination facilitated, but one was able to employ larger quantities for examination, and, by the prolonged shaking and consequently more thorough emulsification, to obtain specimens more representative of the general state of the stool. A second branch of investigation was also undertaken, as the opportunity to add something to the knowledge of the size of cysts in different persons infected was too good to be lost. All who have undertaken the measurement of a large number of cysts of the various parasites will appreciate the tediousness and labour involved in doing this day after day, but it appeared to me that it would be a matter of interest and possible utility to see whether the researches of Matthews and Malins Smith (1917) in Liverpool cases were corroborated by the findings in patients in Jamaica. With a pressure of routine work, and having no trained assistants, I have been able to deal with a limited number of cases only, very limited indeed as compared with the large numbers examined and reported upon by the two authors mentioned, and in putting forward my results, I do so with a feeling of diffidence, since, owing to the limited time and resources at my disposal, I do not intend them as a comparison with their brilliant work. The remarks which follow, however, will serve as a good basis for further investigation which can be continued at any time, and also as a basis for comparison with similar cases in other tropical countries. The method which has been employed is as follows :—Cases for 119 examination were selected from among patients in the hospital to which the laboratory is attached. Those patients who had been admitted for any intestinal condition were excluded. It was desired to find out whether any of those admitted to hospital for complaints other than intestinal were harbouring parasites. Since these patients would leave hospital as soon as they recovered, they would then become healthy carriers, ‘as regards the intestinal parasites present. A certain number of male and female patients were selected, and fresh stools from them were sent to the laboratory on alternate days for three weeks ; on the intermediate days a similar series of an equal number of patients was sent. Two series of patients could thus be dealt with at the same time, or rather during the same period, the stools from one series being brought to the laboratory on Mondays, Wednesdays and Fridays, and those from the other on Tuesdays, Thursdays and Saturdays. Since no specimens were sent up on Sundays, cach case was examined on the first, third, fifth, eighth, tenth, twelfth, fifteenth, seventeenth, and nineteenth days. As nene of the patients showed any signs of intestinal infection they were not under any treatment for the condition even when protozoal infection was detected, so that the natural course and variations could be followed during the period of examination. As Dobell (1917) has stated: ‘A minimum of six examinations per case should be adopted (it is an arbitrary number), no case being regarded as free from infection [he is speaking of Entamoeba histolytica| unless it has been examined six times with negative results. Untreated cases may be examined on any six days convenient to the examiner, since there is no evidence of periodicity in the occurrence of positive or negative examinations. Since the distribution appears to be at random, the chances of finding the infection, if present, are as great for any one day as any other.’ The case J.T. affords an example in which infection with E. histolytica was not detected until the last examination of all; six examinations would have failéd to find this one. The period of three weeks being decided upon, each patient’s stools, if the actions were regular, were examined on nine occasions. Cropper and Row’s concentration method was adopted, and was found to facilitate matters greatly when later we came on to the 120 mensuration of the cysts. There is no need to detail the method; suffice it to say that after concentration three separate specimens were examined from each before the stool was pronounced negative for the day. Examination for helminth ova was also made on the first day that the stools were sent; if these were found, no further search was made for them on subsequent occasions, as the percentage of cases infected with these ova has been given in detail in my reports from Jamaica for several years, totalling over 40,000 specimens. If, however, none were found on the first day, their presence was noted when one came across them later in searching for protozoal cysts, The tables appended show the distribution of the various parasites found, the cases being arranged according to age. It will be seen that the stools from one hundred and two patients have been submitted to examination, of which fifty-three were males and forty-nine females. In twenty-one of the forty-nine female cases the Entamoeba coli was found during these examinations, and in six the &. histolytica. It will be noticed that in only one of these was the latter found without the former. Cysts of Gzardia intestinalis were found alone in four instances, and in combination with other forms in eleven. Chilomastix mesnili was met with seven times and Balantidium. coli once only. Comparing these figures with the numbers found among the fifty-three male cases: E. zstolytica was found in nine, &. colz in twenty-seven, G. inéestinalis in twenty-one (in four of these they were present alone), C. mesnilz was found in six, and B. colz in one. Thus, out of the total of one hundred and two patients whose stools were examined, £. coli was found in forty-eight, or just over 47 per cent. ; E. histolytica in fifteen, or 14°7 per cent. ; G. intestinalis in thirty-six, or 35°29 per cent.; C. mesnzlz in thirteen, or 12°74 per cent.; and B. coli twice. Cercomonas sp. occurred fairly frequently, but no record was kept of this. It is seen in a considerable number of the ordinary routine specimens sent to the laboratory for examination for ankylostome ova. These proportions, when we consider that none of the patients gave any history, at all events recent, of intestinal troubles, are very 12! high, and it was my intention to continue the investigation to see whether subsequent series maintained this high percentage, but my transfer to Hong Kong has prevented this. : Before considering each of these protozoa in greater detail, it will be well to note the criteria made use of in distinguishing the coli and the histolytica cysts ; those of Giardia and Chilomastix need no description. As regards the points relied upon in differentiating coli from histolytica cysts, as is well known, it is the general summation of various characters which enables a definite diagnosis to be made in almost every case; no single character considered alone will suffice for a decision. The points are :— I, size. _ 2. Nuclei: (a) number, (4) character and arrangement of the chromatin. 3. Cytoplasm. 4. Inclusions: (@) chromatoid bodies, (2) vacuoles. 5. Cyst wall. 6. Shape. 1. Szze. The vast majority of Azstolytica cysts have a diameter between 54 and 15m, and of these there are said to be two chief strains met with, namely, ‘small,’ between 64 and gy, with an average diameter of 7°7u, and the ‘ordinary,’ between 10m and 14,4, with an average of 12'6y. Coli cysts vary between wider limits, namely, 11 to 35m, the commonest being between 14m and 22,4, so that difficulties under this head are only likely to arise in differentiating between a large histolytica and a small colz cyst. 2. Nuclei: (a2) Number. In histolytica they may vary from one to four. Malins Smith states that more than half are quadrinucleate, while one-third are uninucleate. In contradistinction to this, more than four-fifths of coli cysts contain eight nuclei, and only about 1O per cent. are quadrinucleate, or less; occasionally a cyst with sixteen nuclei may be met with. It would appear that the consistence of the stool has some influence; thus, it is stated that in loose stools about 60 per cent. of the cysts contain eight nuclei, in semi-formed between 80 and go per cent., and in formed stools between go and 95 per cent. Binucleate cysts are found in about 7 per cent., and quadrinucleate in 3 per cent. 122 (6) Character and arrangement of the nuclear chromatin. The situation of this is, in either case, peripheral, but in Aéstolytzca it consists of smaller granules and is distributed fairly evenly, whereas in coli it is in larger masses or small blocks less evenly distributed, and therefore outlines the nucleus more definitely than is usually the case in histolytica. 3. Cytoplasm. This is of a greenish hue in a fresh specimen of histolytica and is not uniform in appearance, whereas in colz the uniformity is greater, the colour is pale and more of a greyish tint. 4. Inclusions: (a) Chromatoid bodies. These are better seen in fresh saline specimens than in preparations put up with Weigert’s iodine. In Azstolytica they are found in about 30 per cent. of the cysts, are rod-shaped, with blunt, square or rounded ends; whereas they are less commonly seen in colz, only about one in twenty containing them, they are more irregular, are often pointed at the end, or ‘splintered.’ The value of chromatoid bodies in diagnosis is variable, for they may be absent from two-thirds of the cysts. (6) Vacuoles. These are fairly often seen in histolytica, may be multiple, and usually have an ill-defined contour and stain faintly with iodine; in contradistinction the vacuoles of coli are rarely multiple, have a sharply defined edge, and stain more deeply with iodine. 5. Cyst wall. This is usually thinner in Aisfolytica than in col, and the latter sometimes appears to have a thin second contour; these points, however, are far from distinct, and are of minor importance as aiding diagnosis. 6. Shape. Coli is stated to be less frequently asymmetrical than is histolytica, but one comes across cases in which quite a considerable proportion of the colz cysts are asymmetrical; this was particularly noticed in one of the patients in my series. When the majority of these points are in evidence there is no difficulty in diagnosis, but one may meet with cysts which have two or four nuclei, are ill-defined as to colour and uniformity of the cytoplasm (as stated by Matthews and Malins Smith), have no chromatoid bodies, no vacuoles, and not a sufficiently distinctive arrangement of the peripheral chromatin; in such cases certain diagnosis is not possible, but fortunately these are rare. 123 ENTAMOEBA COLI CYSTS These were present, as already stated, in 47 per cent. of the one hundred and two cases examined; there was a slightly larger proportion found among males, viz., twenty-seven out of fifty-three (in the females twenty-one out of forty-nine). Since at least five hundred cases should be examined before any reliable inferences can be drawn as to incidence, it is very probable that further examina- tions would alter this preponderance. As shown in the table, it was the only one found in five cases of the males and in eight of the females; in conjunction with others it was more often met with. In some instances the infection was multiple; thus, in two of the males and one of the females there were Anvkylostoma, Ascaris, Trichiuris, coli, histolytica, Giardia and Chilomastix, without causing any apparent abdominal or intestinal disturbance. * No age seems to be exempt; thus, the cysts were found in a boy ‘of g years of age and in a man of 68 years, in a girl of 13 anda woman of 56. None were found in the five women whose ages exceeded this, probably a mere coincidence. It is interesting to remark that the nearest approach to the proportion of co/z infection found in this series of natives examined (namely, 47 per cent.) was noted by Matthews and Malins Smith in England among asylum patients (45°9 per cent.). For those who are acquainted with the West Indian native, the obvious inference would form a subject of acrimonious if not fruitful debate. Mention has already been made of the irregularity of distribution of the cysts of coli in the faeces, resulting in the finding of several in one specimen and none in others in the same stool; but, by examining larger amounts and concentrating, as-I tried to do, the effects of this irregularity have been reduced. The fact must also be borne in mind that the cysts may appear intermittently and be found in nearly all the preparations made one day, but absent on another in spite of the careful examination of many preparations. Several of the cases given in the table bear out this point. Thus, in No. 52 of the males no cysts were found until the middle of the third week of examination, and in No. 16 they were seen on the first day in conjunction with Aéstolytica, but not again, although the latter was found once subsequently. Other instances of intermittency 124 among the males are Nos. 4, 6, 15, 18, 49. More cases of a single finding of this variety of cyst occurred among the women, namely, Nos. 18, 25, 9, 41, 27, 3, when they were seen at the first, third, fifth and sixth examinations only. In Nos. 42 and 24 they were not seen until the seventh and eighth examinations respectively, and, although there was infection in No. 22 with coli, histolytica, Giardia and Chilomastix, the first-named was found on one occasion only. It has been stated by Dobell that ‘in a series of cases in which the mean number of examinations per case is three, not more than two-thirds, and possibly not more than half, will be detected.’ Taking the first three examinations in this series of cases, there were detected fourteen among the females and eighteen among the males, a total of thirty-two out of the forty-eight in whom these cysts were present—exactly two-thirds. Matthews and Malins Smith found that six examinations give for histolytica about three times the number of positive results obtained by one examination (23 per cent. in place of 7°8 per cent.), and for coli 20°6 per cent. from one examination became 59°8 per cent. by subsequent ones. In my series, five only of the twenty-two positive cases among the females were detected at the first examination (less than one in four), and ten of the men, giving a total of fifteen out of forty-eight in spite of the concentration methods. ENTAMOEBA HISTOLYTICA CYSTS The cysts of Azstolytica are usually more regularly distributed than those of col?, though less so than those of Giardia. A negative record may, as Dobell states, arise either from their having a localised distribution and the part containing them not being submitted to examination, or to their being few in number and so overlooked. He offers an explanation as to the uniformity or localisation of distribu- tion in the site of infection; if the upper part of the large intestine is affected, the faeces here being more or less fluid the cysts are fairly evenly distributed, and when solidification of the faeces occurs lower down the cysts remain scattered through the mass, but, if the lower part is affected, the faeces are already practically solidified and the cysts from the ulcerated surface are thus more superficially distributed 125 and also localised to those parts of the contents which have been in contact with the infected, ulcerated surface. There would seem to be no periodicity in the appearance of cysts of histolytica in the faeces. Owing to the scarcity or to the localised distribution a negative record must not be taken to mean non- infection, unless a large number of examinations are made. Three negative examinations certainly do not warrant a negative report. Thus, only three of the cases among the females were discovered in my series in the first three examinations and not a single one at the first, in spite of the concentration method, showing most emphatically the uselessness of a negative single finding, Among the males only two were found at the first examination, and only six in the first three. Of the men and women together, only nine out of fifteen were discovered in the first three examinations. At the fourth one more was found, another at the fifth, three more at the sixth, and one not until the last time of examination. The tables show also the irregularity in the days of appearance of the cysts of Azs/olyzzca in the faeces; thus, amongst the men they were noted at the first and seventh; first, second, eight and ninth; second, third, fourth, fifth, sixth and ninth; second, fourth and sixth ; second and third only, second and sixth; and so on. Among the women on the second only; the fourth only; the fifth and ninth; the second, third and seventh; the second, fifth, sixth and seventh. These patients were not being treated for the presence of these cysts, for none of them showed any clinical symptoms attributable thereto, and the mere presence of them does not imply amoebic dysentery either present or past, and certainly does not warrant specific treatment. The danger is more, of course, for others, for, though causing no symptoms in the host, they may nevertheless produce acute dysentery in another. In this small series of one hundred and two cases it is seen that a larger number of éstolytica carriers was found amongst the men than the women, in the proportion of three to two. Age appears to have no significance ; thus, of the females one was aged 13, two were 28, one each at 30, 50 and 56 years. Of the men the youngest was 23, then came one at 25, one each at 29, 30, 31 and 32, two at 35 and one at 52 years; the only point which may be mentioned is that seven of the fifteen were found in patients during the third decade. 126 The importance of this infection with histolytica is great. From an examination of a large number of cases, Dobell stated that the incidence of infection does not appear to be conspicuously greater among the cases arriving at the general hospitals for treatment than among those reaching their final depots after such treatment. It would seem, therefore, that the treatment which most of the dysenteric patients had received had not been sufficient to rid them of their infections. This refers to patients who suffer or have suffered from an attack of dysentery. Seeing that the infection is high in my series of patients who had not been abroad (2.e., outside Jamaica) and who showed no clinical symptoms of intestinal mischief, there is every likelihood of greater spread now that large numbers of the contingents sent to the war from that island have returned, and that several of them suffered from dysentery in Egypt and elsewhere. Further, ‘Egypt and its neighbouring lands are notorious centres of amoebic infection.’ Dobell has also shown that passage through an infective area, by affording an opportunity of exposure to infection with fzstolyizca, resulted in the acquisition of infection by a large number of individuals. The incidence of infection in men returning from Egypt and Gallipoli was not conspicuously higher among those who were classified as ‘dysenterics’ than among those invalided for other reasons. This is very applicable to the men who went from Jamaica, for many (if not most) of them served in Egypt, and some, at least, must have brought back infection with them. As a matter of fact, the danger of spread is greater there than at home, for many of those harbouring the cysts appear to be in perfect health, are able to undertake their regular duties, associate freely with their fellowmen, and may spread the disease. Also the duration of carriage in a healthy carrier is not known, and would appear to be almost indefinitely prolonged; though there are intervals when the condition clears up, still this disappearance is only temporary. The two main safeguards against spread are a good sewerage system and abatement of the fly nuisance. Except in Kingston itself, and in parts only of the city, there is no proper sewerage system, the street gutters constituting the sewer for many. As regards the fly question, this is always a difficult problem in the tropics, and since we know that flies can take up /?stolytica cysts, pass them unchanged through their bodies and so infect food, the danger is clearly a very real one. 127 Wenyon and O’Connor’s (1917) remarks on the fly question may be briefly quoted here :— 1. Flies readily take up encysted and other forms of protozoa into their intestines. 2. Encysted forms remain in the intestines of the fly as long as there is any faecal matter. If prevented from feeding they may retain the cysts for as long as forty-two hours; if feeding is allowed, they do not retain the cysts as long, and the flies may deposit material (and with it cysts) ingested only five minutes previously. 3. Cysts do not degenerate to any extent in the intestines of the fly, but readily pass unaltered. GIARDIA INTESTINALIS There is no proof that Giardia is pathogenic, at any rate in adults. It has been found in a considerable number of instances in this series, sometimes in a free state, sometimes encysted and in very large numbers. Its presence has been noted in thirty-six of the one hundred and two cases, or 35°29 per cent.; of these twenty-one were amongst the fifty-three males and fifteen amongst the forty-nine females. This, again, is a higher percentage than was found in cases recorded by Dobell, Wenyon and O’Connor. The first-named found 27°3 per cent. with a minimum of six examinations, but states that this was probably not more than three-fourths of the real number. This parasite is usually found more frequently in patients suffering from diarrhoea, but it is probably nearer the truth to say that the diarrhoea or loose action is the ‘cause’ of the detection of the infection than vice-versa, that the infection is the ‘ cause’ of the diarrhoea. Two reasons may be offered to account for the high percentage of infection in my series; firstly, that in order to ensure the procuring of specimens many of the patients were given salines, thus producing a loose action, and secondly, the concentration method allowed of the discovery of the organism in cases where they would otherwise have been so few as to be overlooked. In reports of examinations carried out in England, it has usually been found that Giavdza was much more common in children, so 128 much so that Matthews and Malins Smith suggest that this flagellate is mainly a parasite of children and that it may disappear from the intestine in course of time. In my series it was found in all cases examined (six) below the age of 15 years, but was also present in others of 20, 56, and 60 years of age (five out of eight in the sixth decade) ; among the females it was found in two of the three under 15 years of age, but also in seven out of seventeen between 21 and 30 years, and in one each at 37, 48 and 56 years. CHILOMASTIX MESNILI Cysts of this parasite were found six times among the males and seven among the females, giving a total percentage of 12°74. In some instances they were exceedingly numerous, occurring not only in every field, but sometimes more than one in the field, the concen- tration method accounting in part for this. They are probably of no pathogenic significance. Lastly, it may be mentioned that Balantidium coli was found twice: in a woman of 56 and a man of 22 years of age, in the latter as the only parasite detected , in the former associated with &. colt. The appended tables give succinctly the chief points in connection with this investigation. The patients have been arranged in order of age for ease of reference. REFERENCES Cropper, J. W., and Row, R. W. H. (1917). A method of concentrating Entamoeba cysts in stools. Lancet, Vol. CXC, pp. 179-182. Dosett, Crirrorp (1917). | Amoebic Dysentery and the Protozoological Investigation of Cases and Carriers. Med. Res. Committee, Special Report Series, No. 4. Smitn, A. Matins, and Matruews, J. R. (1917). Further Records of the Occurence of Intestinal Protozoa in non-dysenteric cases. Ann. Trop. Med. & Parasitol., Vol. 1X, pp- 183-193. Wenyon, C. M., and O'Connor, F. W. (1917). 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GbE = = = WD HD = ‘DH’ x ‘OH = ae + + 4+ ae de ise oi orp “+ qoearqeg | SE WL | ££ = > = ‘H = a cd = — - + s+ (Teo9090u03) siytatjounfuog | SE ‘da | ze = = = pa = = ~ ne = ps = con ects ++ guog yo sisoss0Ny | SE ‘of } 16 — = any = = ao =< se = ae = eee ese ore “+ BIIRTEYAL + ‘a‘o of 0'9 3:9 a 7) Do) <4 es 7) ies ay, do a tee aes “+ — . - ‘ ‘ . -® ¢ Ae 4 } 5 s v : “ee i a d 2 ‘ ey am ser . ‘ , va . = a a . . : € a) { 173 A CASE OF SUSPECTED LEPROSY BY J. W. W. STEPHENS AND S. ADLER (Received for publication 22 June, 1921) PLaTE XVI X.Y., age 16. Born in England, 1904. Has resided in Capetown, S. Africa, from 1905 until September, 1920. History of Illness :-— April, 1920. Wasting noticed on dorsal side of left hand between Ist and 2nd metacarpals, and a stinging sensation in the little finger. June, 1920. Three spots noticed on left side of face and brownish patches on both eyebrows. Later brownish patches noticed on left forearm and hand, and on both lower limbs. August, 1920. Noticed deformity and impairment of movement in left hand. Examination of the patient :— 1. A number of dry, scaly, brownish, thickened areas in part cir- cumscribed up to three-eights of an inch in diameter, in part somewhat diffuse. None of these areas was anaesthetic except for those involved in the areas of anaesthesia, to be described later. The distribution of these is shown approximately in figs. 1 and 2, 2. A condition of claw hand (left). The actual condition can be appreciated from the photographs (Plate XVI). 3. Thickening of the left ulnar nerve which in the ulnar groove was about three times as thick as on the right side. The left internal cutaneous nerve was palpable and about a quarter of an inch thick. 4. The left forearm and hand were anaesthetic over an area shown approximately in figs. and 2, 5. The left hand was colder to the touch than the right. 174 CY, Fics. 1 and 2. The small circles indicate the affected circumscribed areas, the stippling the diffuse areas, and the oblique lines the anaesthetic areas. 175 From 30.12.20 to 14.4.21 during his stay in the ward there was no rise of temperature. 27.1.21. The Wasserman reaction was completely negative. On 7.2.21 and 21.2.21 the patient burned his left hand on the hot water-pipes of the ward without being aware of it. Examination for Leprosy Bacilli :-— I. One of the affected areas on the left forearm was excised at the London School of Tropical Medicine, November, 1920, and examined by Dr. Low for leprosy bacilli with negative result. 2. Numerous scrapings and punctures of the various skin lesions, e.g., on the eyebrows, face, and of the nasal mucosa, were negative. 3. 2 c.c. of blood from a vein were haemolysed and centrifuged ; the deposit was examined for leprosy bacilli with negative result. 4. A piece of the internal cutaneous nerve was excised, and sections and smears were stained for leprosy bacilli with negative result. The excised nerve was examined for us histologically by Professor E. Glynn, who reported that ‘there are three caseous areas apparently corresponding to large nerve bundles, surrounded by fibrous tissue in process of formation. No nerve fibres were seen. The appearance is quite consistent with fibro-caseous leprosy.’ On re-examination of the patient on 14.6.21 the only change noticed was a disappearance of affected areas on the dorsum of the hand and a diminution in extent of some of the areas on the face. In view of the fact that no evidence of the presence of leprosy bacilli was obtained no treatment was adopted. 176 EXPLANATION OF PLATE XVI Fig. 1. Ventral aspect of right and left hands. The left hand shows (a) its smaller size; (6) ill-defined cutaneous furrows ; (c) wasting of the thenar and hypothenar eminences—flatness of the hand; (d) flexion of the interphalangeal joints of the 2nd, 3rd, 4th, and 5th fingers ; (e) indication of the thickened rough skin areas at the base of 3rd, 4th, and 5th fingers ; (f) the scar of a burn at the base of the hand on the ulnar side. Fig. 2. Lateral view of the right and left hands. The left hands shows (a) wasting in the interspace between the 1st and 2nd meta- carpal bones; (5) extension of the metacarpo-phalangeal joints and flexion at the interphalangeal joints ; (c) wasting of the forearm. Annals Trop. Med. & Parasitol., Vol. XV PLATE XVI Photographs by Miss M. Brown C. Tinling & Co., Ltd., Imp 77 OBSERVATIONS ON THE CERATOPOGONINE MIDGES OF THE GOLD COAST WITH DESCRIPTIONS OF | NEW SPECIES PART IV BY HENRY F. CARTER A. INGRAM AND J. We S. MACFIE (Received for publication 7 April, 1921) Genus DASVHELEA, Kieff. Ceratopogon, Wtz. (pro parte) (nec. Mg. Edws.). © Linnea Entomologicae.- Vol. VI, » 1852. Culicoides, Kieff. et auct. (pro parte) (nec. Latr.). Dasyhelea, Kieft. Bull. Soc. Hist. Nat. Med. Vol. III, 1911. Prokempia, Kieff. Rec. Ind. Mus. Vol. IX, 1913. Pseudoculicotdes, Mall. Bull. Ill. Sta. Lab. Nat. Hist. Vol. X, 1915. Dasyhelia, Brunetti. Rec. Ind. Mus. Vol. XVII, 1920. This genus is closely related to Culccoides, Latr. It was erected by Kieffer in 1911 for the Indian species D. halophila, and subsequently (1913) divided by him into three sub-genera— Dasyhelea (sens. stric.), Prokempia and Kempia—according to the nature of the wing hairs and the empodium. Later (1917) the last two sub-genera were given generic rank by this author, who then associated Prokempia with Dasyhelea but removed Kempfia to the Atrichopogon group. Still later (1919) Kieffer again placed Prokempia as a sub-genus of Dasyhelea. Prokempia apparently differs from Dasyhelea only in the absence or relative scarcity of the longer wing-hairs, and was separated for this reason; but although the density of arrangement of these hairs is much more uniform in Dasyhelea than in Culicoides, this character cannot, in our opinion, be considered of more than specific value. 178 Malloch, in 1915, founded the genus Pseudoculicoides for Ceratopogon mutabilis, Cog., especially distinguishing it from Culicoides by the absence of thoracic cavities and the antennal structure of the male. The description and figures of this species given by Malloch indicate that it is a typical Dasyhelea, and Kieffer (1919) accordingly placed Pseudoculicoides as a synonym of the latter. Malloch, however, made no reference to the eyes—the hairiness of which was given by Kieffer as one of the diagnostic characters of the genus—either in his generic definition or specific description; but Mr. F. W. Edwards, in a private communication, informs us that the eyes of specimens (named by Malloch) of C . mutabilis in the British Museum collection are shortly but distinctly hairy, and therefore Kieffer’s decision in regard to this question must be accepted. EXTERNAL MorPuHotocr. The detailed anatomy of Dasyhelea differs from that of Culicoides (see Part II of this study, 1920) as follows :— ApuLts. Head. Eyes in both sexes densely clothed with microscopic hairs. Mouth-parts (fig. 1). Proboscis somewhat shorter than that fig : pA c d Fic. 1. Mouth parts of D. pallidibalter(Q): a—labrum; 6—maxilla; c—mandible ; d—hypopharynx. (xX 490 circa.) of Culicoides, the component organs, except the labium, differently formed and apparently not adapted for piercing in either sex. The labrum is strongly chitinised, broad at the base gradually tapering 179 to a rounded, hairy apex. The hypopharynx is similar in shape to the labrum, but tapers more rapidly and ends in a pointed apex; the distal fourth bears numerous hair-like marginal processes. The mandibles and maxillae are poorly developed and devoid of teeth; the former are closely applied to (and not easily separated from) the labrum and hypopharynx, and are enclosed in membranous sheaths which extend considerably beyond their extremities ; the maxillae are rudimentary and are reduced to small, thinly chitinised, blade-like structures. The palpi in both sexes appear to be composed of four segments owing to the first being very rudimentary and often almost indistinguishable; the third segment is usually distinctly longer than the fourth or fifth and is scarcely swollen, with the minute sensory hairs scattered over the inner surface from the base almost to the apex—not ccncentrated in a relatively deep and sharply defined depression, The above description of the mouth-parts is based upon dissections of two of the new species (D. pallidihalter and D. flava) described herein, and of the European D. obscura, Wtz. The structure of the various organs is apparently the same in males and females, and, in our opinion, is such that these midges would be incapable of piercing the skin. Malloch (1915), however, states that D. (Pseudoculicoides) griseus, Coq., ‘was taken biting, on the bank ~ of Sangamon River,’ and that D. cimctus, Coq., has been ‘ recorded as biting human beings.’ In Accra, specimens of Dasyhelea spp. occasionally alighted on the arm, but, so far as observed, they made no attempt to bite; possibly, therefore, the statement regarding D. griseus may have been due to a similar occurrence, and may subsequently require modification. At least, until the feeding habits of the midges of this genus have been carefully studied, the above statement must be regarded with reserve. Antennae: segments of the flagellum in the female shorter and broader than. in Cudicotdes, gradually lengthening towards the apex, but with the apical ones not distinctly differentiated from the basal ; last four segments (twelve to fifteen) in the male, elongate, the twelfth to fourteenth inclusive cylindrical, binodose,* the whorls of * The apical node is usually less well-developed than the basal node, and the hairs are shorter and more slender ; but there is distinct variation in this respect, and in D. flava, sp. n. (see p. 196), the apical whorl is obsolete. 180 hairs arising from nodal rings situated on the basal and apical thirds of each segment. In both sexes, but more conspicuously in the males, the segments of the. flagellum exhibit a tesselated or sculptured appearance, due to the presence of small chitinous plates, from the upper portion of each of which a hair arises; on the basal segments (four to eleven) in both sexes this sculpturing appears to be limited to the proximal portions (¢.e., the area below the whorls) of each, where the plates are sharply defined and radially arranged ; on the distal segments the plates are arranged as shown in fig. 2. Fic. 2. Terminal segments of antennae of a—D. fusciformis (gd) and b—D. inconspicuosa (Q). (X 475 circa.) Thorax without anterior or posterior pits or depressions; scutellum with a varying number of strong marginal or sub-marginal bristles and usually a few (one to six) short hairs. Wings hyaline, unspotted; the venation very similar to that of Culicoides, but with the first and third veins less distinctly separated, so that normally only one interspace can be distinguished ; in some species these veins appear to be completely fused. Legs. Tibiae without apical spurs, but the fore and hind pairs with oblique or transverse rows of bristles distally; first tarsal 181 segments of all legs sub-equal, those of the middle legs not appreciably elongated. Claws small, equal, simple in the female, divided at the tips in the male; in both sexes the inner margin may be somewhat prominent, tooth-like, at the base, and agulate near the middle. : Abdomen. Dorsum covered with minute flattened spines; segments one to seven each with two small, clear, thinly chitinised, rounded areas. Spermatheca (fig. 9, 4-f) single, heavily or moderately chitinised, usually sub-spherical, the commencement of the duct sometimes chitinised for a considerable distance. External genitalia of the male.* Ninth segment: tergite, apical lobe-like processes well developed, often projecting posteriorly beyond the tergite and sometimes bearing a median dorsal triangular chitinised plate; sternite not excavated, sometimes produced posteriorly in the middle line (c.f. figs. 10, 11). Forceps: side-pieces similar to those of Culicoides ; claspers simple or divided, if the latter the branches variously formed, dissimilar. Harfes with the proximal portions greatly developed, highly chitinised, and in some species connected medially to form an uninterrupted, or almost uninterrupted, transverse bar. The distal portions show varying degrees of development and chitinisation, and in all the species, except one, examined by us, the distal portion—bent sharply and directed posteriorly—of only one harpe is developed, thus causing asymmetry; in the exception referred to (D. similis, sp.n.) the distal portions of both harpes appear to be developed, but are fused in the middle line forming a conspicuous median structure. Aedoeagus strongly chitinised, usually broad but of variable form. * In regard to the nomenclature of the hypopygial structures adopted in this and previous studies, Parts II and III (1920-1921), on the Ceratopogonine midges of the Gold Coast, Mr. F. W. Edwards writes us as follows :—‘ I now think that your harpes are the same structures which de Meijere has called gonapophyses in the Limnobiidae, and I was probably right in homologising them with the parameres of Culicidae. There is one difference, at first sight fundamental, in that in Culicoides and Limnobiidae (c.g., Eriocera) the structures are dorsal to the aedoeagus, adjoining the tenth sternite (which I consider to be represented by your spicular strip of membrane), whereas in the mosquitoes they are morphologically ventral to the aedoeagus. However, it would seem likely that this is connected with the atrophy of the anal chitinisations in Culicoides and the Limnobiidae, the parameres moving dorsally and fusing in the mid-dorsal line in order to replace, in function, the tenth sternite. In other insects the parameres are said to be normally lateral in position, so that it is easy to understand that they might pass either dorsally or ventrally. In any case the important idea to lay hold of is that the parameres form with the basal plate the first (morphologically innermost) ring of the genital tube, the second ring being the mesosome which is more or less invaginated into the first. In Culicoides I believe the structure you call the aedoeagus is really the basal plate only, and that there is no mesosome. Properly speaking the term aedoeagus should include in this case also the “harpes.”” The two small chitinous appendages of the ninth tergite, of course, represent the tenth tergite.’ 182 PupA. The pupae of Dasyhelea much resemble those of Culicoides, the chief point of difference being in the reduction of the first and elongation of the second abdominal segments, the shape and structure of the respiratory trumpets and the arrangement of tubercles on the body. The structure of the trumpets, as shown in the species examined by us, is of especial interest since forms occur which represent transitional stages between the simple type, as exemplified by Culicoides, and the complex type as found in at least some species of Forcipomyia (e.g., F. ingrami, Cart., 1919). Such forms are shown in fig. 3, 6-d; the last type figured evidently represents ARC. d ee Fic. 3. Pupal trumpets of L—D. flaviformis ; c—D. ingrofusca ; d—D. fusca. (b—c X 360 circa; d X 220.) a stage in development immediately preceding that occurring in Forcipomyia, for by a simple folding back of the distal leaf-hke portion an organ very similar in structure to the trumpet of F.. ingrami could be produced. 183 The chief differences in the arrangement of the body tubercles ‘is as follows:—Cephalothorax: anterior dorsal tubercles absent, anterior dorso-median tubercles present (as in Szilobezzia), dorsal, postero-dorsal, and ventro-median tubercles absent but sometimes represented by minute hairs; the dorsal surface of the thoracic portion is thus entirely devoid of distinct tubercles, but is, on the other hand, usually pigmented or striated in circumscribed areas. Abdomen: tubercles flattened and flange-like, the dorsal antero- submarginal tubercles small or rudimentary, one only on each side; the postero-marginal tubercles are situated, as in S/ilobezzia, near the middle transverse line of each segment, around which they form an almost continuous (interrupted in the middle line of the dorsal and ventral surfaces) band ; on segments three to seven they are nine in number, of which the three central ones (ventro-lateral tubercles) are the most highly developed. In some species small, rounded, pigmented areas, arranged in the form of a triangle, are present on both dorsal and ventral surfaces. The terminal processes are broad and flattened, and are directed dorsally and laterally; at the base of each are two prominent tubercles bearing hairs and spines. LarvA. The mature larva differs from that of Culicoides chiefly in regard to the size and shape of the head, the position of ~ the eyes and in the possession of a small hooked pseudopod at the posterior extremity of the body (fig. 9, g). The ead is distinctly larger (almost twice as large as in Culicoides) and tapers more strongly towards the apex, being obovate rather than sub-conical; it bears minute hairs only. The eyes are reniform and are situated far forwards, near the anterior third. The mouth-parts are similar in structure to those of Culicoides, but, in the three West African species observed, appear to be rather more highly developed in so far as the hypopharynx is larger and more complex, the mandibles are tridentate and the labium is more highly chitinised and more strongly serrated. The body is elongate and scantily clothed with very minute hairs; the anal pseudopod is retractile, small and membranous, and bears two dorsal and two ventral groups of small but strongly chitinised, recurved hooks which are directed anteriorly. The anal. papillae are short, bifid, with slightly blunted extremities. 184 Speciric DESCRIPTIONS. Dasyhelea pallidthalter, sp. n. MEASUREMENTS. Female. Mate. Length of body* ... pe es 560 so Bpedbaice arin I°3 mm. Length of wing... un a 28 ... o8 mm. o-8 mm. Greatest breadth of wing ... 0-3 mm. 0-3 mm. ° Head dark brown, clothed with dark hairs. Eyes narrowly separate in both sexes. Proboscis and palpi brown, the latter with the third segment uniformly and slightly swollen and almost twice as long as the fifth; the second, fourth and fifth short, sub-equal. Antennae dark brown with dark brown hairs; in the female the segments of the flagellum are sub-spherical at the base, gradually lengthening, becoming oval, towards the apex, the fourth to eleventh each bearing a pair of large, strong, bluntly rounded spines; in the male, segments four to eleven are sub-spherical to ovoid in shape, the length from two-thirds to one and one-third times the greatest breadth, segments twelve to fifteen are elongate, about three times as long as wide, the last broader and pointed distally, the twelfth to fourteenth sub-cylindrical, each distinctly binodose. Thorax dark brown with small yellowish-brown areas in the humeral angles and over the roots of the wings. Scutellum dark brown, becoming brownish-yellow centrally, bearing four centro-marginal and two lateral bristles, and a median transverse row of about six short hairs. Post-scutellum and pleurae dark brown. W2zmgs in the female well clothed with decumbent hairs which extend almost to the base between the fourth and fifth veins and are numerous in the anal angle and fork of the fifth vein. Costa and first and third longitudinal veins not extending to the middle of the anterior border, terminating above or slightly beyond the bifurcation of the fifth vein and enclosing a small but distinct cell or interspace; in the male less hairy (no hairs at the base of the wing, and very few, if any, in the anal angle or fork of the fifth vein), the costa terminating slightly before the bifurcation of the fifth vein. Halteres white, or almost white. Zegs uniformly dark brown. Claws in both sexes with the inner margin at the base somewhat produced, and near the middle with a small, angulate prominence. Addumen: dorsum * In all cases taken from the anterior margin of the thorax to the tip of the abdomen of specimens mounted in carbolic. 185 dark brown, sparsely clothed with dark brown hairs, venter rather paler brown. Spermatheca highly chitinised, spherical (diameter 50) with a conical posterior extension (length about 20) leading to the duct. Hyrorycium (fig. 4). Nnth segment: tergite short and broad, relatively strongly chitinised and sparsely clothed with moderately long, strong hairs, posterior margin straight, produced on each side into a small, rounded, hairy knob, apical lobe-like processes well developed, often extending as a centrally indented membrane considerably beyond the extremity of the tergite; sternite broad, Fic. 4. D. pallidibalter, sp.n., male hypopygium (ventral view). X 400 circa. i produced centrally and posteriorly into a relatively large, more or less oval, lobe-like process. Forceps: side-pieces well chitinised and with long hairs, as in Culzcoides; claspers long and slender, rather strongly chitinised, the basal half pubescent with a few strong hairs, the apex pointed, slightly depressed, bearing two or three short, delicate hairs. Harfes as shown in fig. 4, the proximal portions very strongly chitinised, the distal portion—only that of the left* harpe is developed—very feebly chitinised, narrow and bent almost at a right angle to the proximal portion. Aedoeagus somewhat V-shaped, relatively short, the lateral arms with outwardly directed, * In this and all subsequent species in which the harpes are not symmetrically developed, the side of the insect from which the distal portion arises is stated. The figures of the hypopygium, however, show the ventral aspect and therefore in them the right side of the insect lies to the left of the reader. 186 pointed, beak-like, distal processes and connected anteriorly by a ~ slightly curved, centrally thickened chitinous bar. Pupa. Length 1.9 mm. Integument pale brownish-yellow, not infuscated in any part or heavily shagreened except on the operculum and on the posterior abdominal segments. Resfiratory trumpets narrow and strongly curved, almost semi-circular; length about o'3 mm. The trumpets are obliquely and irregularly ringed, the tracheal trunk in each gives off a number of short lateral branches, which lead to small elevations on the surface, and terminates distally in five or six short, blunt processes. Cephalothorax: dorsum of thoracic portion not pigmented, smooth except on anterior border, and with a few puckered or striated areas. Anterior marginal tubercle large, shagreened, bearing a short bristle; dorso-lateral tubercle rounded, smooth, bearing two or three small hairs, anterior dorso- median tubercle small, bearing a short hair; ventro-lateral tubercle moderately large, rounded, bearing a short spine and a short hair; ventro-median tubercle represented by two minute hairs. Between the anterior marginal tubercles are two (one dorsal and one ventral) rounded, unarmed median humps. Dorsal tubercles absent, but anteriorly represented by one or two minute hairs. Addomen: integument reticulated and slightly shagreened at the sides and base of each segment, more heavily shagreened on the distal segments, the last entirely shagreened. Tubercles arranged as mentioned on page 183, the antero-submarginal tubercle small armed with a short spine, the dorsal and ventral postero-marginal tubercles apparently unarmed, low, broad and flange-like, the ventro-lateral postero-matginal tubercles larger and conical, the middle bearing a short hair, the others a short spine. Terminal processes broad and flattened, directed dorsally and laterally, each with two posterior tubercles, bearing respectively a hair and a spine. Larva. Length about 3 mm. Head obovate, convex dorsally, flattened ventrally; length o'2 mm., greatest breadth o'112 mm. Hairs scanty, minute. Mental plate crescentic, moderately chitinised, apparently bearing nine teeth on each side, the last large and broadly rounded, the others small, narrowly rounded and closely apposed. Mandibles large, highly chitinised, the distal portion bearing three large teeth on the inner side. Hypopharyngeal sclerite very highly chitinised posteriorly, the posterior portion 187 armed with three rows of teeth and with a delicate, serrated, fringe- like expansion on each side. Body scantily clothed with minute hairs, the last segment with apparently eight pairs of very small hairs near the extremity. Posterior pseudopod (fig. 9, g) small, the dorsal groups each consisting of two hooks, the ventral groups each of four hooks. HABITAT: Nsawam, Gold Coast. Reared from rotting material collected from the base of a banana plant; March to May, 1920. Dasyhelea fusciscutellata, sp.n. MeasureMENTS. Female. Male. - Length of body (two specimens) ... ae .. TOmm. I-I mm. Length of wing tee = ae re .. O8 mm. o:8 mm. Greatest breadth of wing ... ... a Seng O74) 0TH o-3 mm. Head dark brown with almost black hairs. Eyes narrowly separate in the female, very narrowly separate in the male. Clypeus, proboscis and palpi brown; third segment of the latter cylindrical, slightly and uniformly swollen, about as long as the fourth and fifth segments together.- Atennae brown with black hairs and, except on the last five segments, long spines; in the female the segments ovoid and sub-equal in size gradually elongating towards the apex, but the thirteenth and fourteenth segments slightly longer than the eleventh and twelfth, last segment the longest, not produced distally into a stylet: in the male, segments four to eleven spheroidal, gradually becoming narrower and more elongate apically, the last four segments longer, the twelfth and thirteenth sub-equal in length, each slightly longer than the fourteenth, the last segment about as long as the penultimate and without a terminal stylet, segments twelve to fourteen distinctly binodose. Thorax dark brown with black hairs, the anterior lateral angles sometimes of a pale brown colour. Scutellum dark brown with four centro-marginal bristles, two lateral bristles, and two small hairs. Postscutellum and pleurae dark brown. Waéings sparsely clothed with decumbent hairs which extend basally between the fourth and fifth veins beyond the anterior cross vein. In the female the costa terminates near the middle of the anterior border but extends well beyond the bifurcation of the fifth vein, and the first and third veins form distally a somewhat indefinite cell; in the male, the costa ends before the bifurcation of 188 the fifth vein and the first and third longitudinal veins form a distinct cell. Halteres with tawny-yellow knobs and dark brown stems. Legs brown, uniformly coloured; claws with a basal hairy extension, of the female simple, each with a slight sub-central projection, of the male bifid distally, each with a distinct sub-central projection. Adédomen dark brown with black hairs. Spermatheca (fig. 9, 7) heavily chitinised, somewhat reniform, the anterior portion ovoid (length 46, breadth 49), the posterior portion (length 18) narrower and sharply curved. : Hyrorycium (fig. 5). Nzuxth segment: tergite broad, slightly tapering posteriorly and bearing a few moderately long hairs dorsally, posterior margin without a central notch and with the lateral finger-like processes small, apical lobe-like processes well Fic. 5. D. fusciscutellata, sp.n., male hypopygium {ventral view). X 400 circa. developed and with a median triangular dorsal chitinised plate between them; sternite with a somewhat rectangular median posterior projection. Forceps: side-pieces strongly chitinised, with long hairs; claspers well chitinised, the basal halves pubescent and bearing one or two relatively stout hairs, the distal extremities shghtly depressed. Harges: basal portions strongly chitinised, broad laterally, narrowing towards the middle line; from the left basal portion a feebly chitinised, slightly curved, pointed, blade-like process projects posteriorly as far as the middle of the side-pieces. Aedoeagus : median transverse basal portion not very broad, heavily chitinised and slightly twisted and bevelled laterally, lateral portion 189 strongly chitinised, the distal extremities hook-like and directed dorsally. HaBitaT: Nsawam, Gold Coast. Reared from materials collected from the bases of banana plants, March, 1920. Dasyhelea similis, sp. 1. MEeasurEMENTS. Female. Male. Length of body... a phen Hr egae eons r mis I-o mm. Length of wing + a: os oad -. O7 Mm. o-7 mm. Greatest breadth of wing ... ae a .. 0-3 mm. o-2 mm. Head dark brown with dark brown hairs. Eyes narrowly separate in both sexes. Clypeus, proboscis and palpi brown, the third palpal segment slightly but distinctly swollen at the base and attenuated distally, in the female about as long as the fourth and fifth segments together, in the male relatively shorter, about as long as the fifth. Ax/ennae brown with brownish-black hairs and, except on the last five segments, long, clear spines; in the female the segments, with the exception of the last, which is the longest but carries no stylet, are all sub-equal in size and ovoid in shape, any one of the apical segments (twelve to fourteen) being, if anything, slightly shorter than any one of the five (seven to eleven) preceding ; in the male, segments four to eleven spheroidal gradually becoming narrower and longer towards the apex, the last four segments elongate, twelfth and thirteenth sub-equal, longer than the others, twelfth to fourteenth distinctly binodose. Thorax dark brown, the anterior lateral angles sometimes paler brown. Scutellum dark brown with four* centro-marginal and two lateral bristles, and two or more small hairs. Post-scutellum and pleurae dark brown. Wings sparsely clothed with decumbent hairs which scarcely extend basally beyond the cross-vein; in the male the costa extends slightly beyond the middle of the wing, its end lying above the fork of the fifth vein, and the first and third veins form a distinct cell; in the female the costa extends beyond the middle of the wing and terminates distally to the fork of the fifth vein, forming with the first and third vein a somewhat indefinite cell. Halteres with pale yellow knobs and dark brown stems. Legs uniformly brown. Female claws simple, male claws bifid with a slight indication of a sub-central projection; in both sexes with a hairy extension’ * The two middle centro-marginal bristles may be replaced by a single central bristle in this and other species of Dasyhelea. 190 at the base. Addomen dark brown clothed with dark hairs. spermatheca (fig. 9, e) sub-spherical (diameter 46), moderately chitinised, the duct scarcely chitinised at its commencement. Hyporycium (fig. 6). Ninth segment: tergite scarcely tapering posteriorly, the margin straight with rudimentary finger-like processes each bearing two setae, the apical lobe-like processes large ; sternite not produced posteriorly. Forceps: side-pieces clothed with long hairs, claspers strongly chitinised, the basal halves pubescent, the apical halves bearing one or two short hairs near their distal extremities which are depressed. Harges: basal portions curved, ribbon-like, symmetrical, joined medially, forming a 490 —_— Tic. 6. D. similis, sp.n., male hypopygium (ventral view). X 400 circa. transverse bar from the centre of which a heavily chitinised structure (representing the fused distal portions) extends posteriorly and ends in arather blunt point. Aedoeagus: median transverse basal portion strongly chitinised, narrow, somewhat thickened in the middle (triangular in some views), and but slightly curved, lateral portion terminating in moderately sharp points and bearing dorsally a strongly developed barb which projects transversely and is curved in an anterior direction. HaBitaT: Nsawam, Gold Coast. Reared from rotting material collected from the bases of banana plants, March, 1920. D. similis closely resembles D. fusciscutellata but is of a rather lighter colour, generally with hghter yellow halteres; the species may be readily distinguished by the form of the spermatheca in the females and the structure of the hypopygium in the males. IgI Dasyhelea luteoscutellata, sp. n. MEASUREMENTS. Female. Length of body (one specimen)... foe ads ae es. Tel mm. Length of wing... Bs te #c B5 a ... O8 mm. Greatest breadth of wing ... sf es on as .-- 0°73 mm. Head dark brown with dark hairs. Eyes narrowly separate. Clypeus, proboscis and palpi brown, the third palpal segment longest, slightly swollen basally, the fourth and fifth segments together longer than the third. Azztennae brown with dark brown hairs and long spines, curved and somewhat hair-like, on segments four to ten, inclusive; the latter segments sub-spherical to ovoid, elongating towards the apex, the terminal five segments distinctly more elongate, together longer than segments three to ten united, the last segment without a stylet. Toray dark brown with dark hairs. Scutellum bright yellow with four centro-marginal and two lateral bristles, and a small central hair. Post-scutellum and pleurae dark brown. W2zzgs similar to those of D. pallidihalter. Halteres with white knobs and dark brown stems. Legs brown, uniformly coloured. Claws simple without a sub-central prominence. Abdomen dark brown clothed with dark brown hairs. Spermatheca sub-spherical (diameter 43); the duct chitinised for a short distance only. HABITAT: Nsawam, Gold Coast. Reared from material collected from the base of a banana plant; the pupa and larva were not identified. This species is a small, dark brown or almost black midge, resembling D. pallidihalter but with the scutellum entirely bright yellow, not darkened laterally; the long spines on the basal segments of the antennae are more slender and hair-like than in D. pallidthalter. Dasyhelea inconspicuosa, sp. n. MEASUREMENTS. Female. “Male. Length of body (two specimens) ... a ome OM. I'l mm. Length of wing... ai oe te .. O7 mm. o-9 mm. Greatest breadth of wing ... et oa .. Of Mmm, o-5 mm. Head dark brown. Eyes broadly contiguous in the female, narrowly separate in the male. Proboscis and palpi brown, the 192 latter relatively long, the third segment elongate, cylindrical, as long as the fourth and fifth segments together, the second segment about half the length of the third. Aztennae brown with dark brown hairs; in the female, segments four and five sub-spherical, six to fifteen oval, gradually becoming longer towards the apex, the last segment broad without a stylet (fig. 2, 6); in the male, segments four to eleven spheroidal to ovoid, segments twelve to fifteen elongate, the twelfth and thirteenth sub-equal, longer than the fourteenth and fifteenth, the last segment rather shorter than the fourteenth, without a stylet. Thorax dark brown with dark hairs. Scutellum yellowish brown with four centro-marginal and two lateral bristles in both sexes, small hairs absent. Post-scutellum and pleurae dark brown. Wings densely clothed with decumbent hairs, which in the female extend to the base between the fourth and fifth veins. In the female the costa extends to about the middle of the anterior margin, its extremity distal to the bifurcation of the fifth vei, forming with the first and third veins a narrow, slit-like cell; in the male the costa scarcely reaches the middle of the anterior margin and terminates above the bifurcation of the fifth vein. Halteres with pale brown knobs and dark brown stems. Legs brown. Male claws bifid at their extremities and with a minute sub-basal hairy extension on their inner sides; female claws simple, with a sub-basal extension similar to that of the male, but smaller. Addomen: dorsum dark brown with brown hairs, venter paler brown. Spermatheca strongly chitinised, spherical (diameter 32), the chitinised portion of the duct straight (length 5,). Hyporycium (fig. 7). Ninth segment: tergite scarcely tapering towards its posterior extremity and scantily clothed with long, stout hairs, the finger-like processes moderately small and somewhat pointed, each bearing a relatively long hair near the apex and four near the base, apical lobe-like processes well developed and bearing between them a small triangular chitinised plate (not shown in the figure), sternite prolonged centrally and posteriorly into a long narrow process which covers the aedoeagus and ends in what looks like, but actually is not a strongly chitinised spine. Forceps: side-pieces well chitinised, with very long hairs, claspers with the basal halves pubescent and the tips depressed, and bearing a few minute hairs. Harfes: basal portions heavily chitinised, broad 195 laterally and somewhat L-shaped; distal portion of the left harpe developed but small, slender and feebly chitinised. Aedoeagus: median transverse proximal portion slightly curved and tapering laterally; lateral portions look as if they were twisted, apparently each composed of two plates, between them a tapering median gutter-like process with a blunt end (not shown in the figure); the junction between the plates on each side is very thinly chitinised. PupaA: Length (two specimens) 2 mm.; dorsum of the cephalo- thorax somewhat infuscated anteriorly, the anterior margin and the Fic. 7. D. inconspicuosa, sp.n., male hypopygium (ventral view), X 400 circa. posterior end of the abdomen conspicuously shagreened. ResPira- tory trumpets: \ength about o'2 mm.; curved, and conspicuously ringed except at the extremities. The main tracheal trunk gives off laterally, at fairly regular intervals, a few processes which open on elevations slightly larger than the ordinary corrugations of the trumpet, and apically about eight short blunt processes arranged in a fan-like manner. Cephalothorax without strongly armed tubercles. Anterior marginal tubercles divergent, dark coloured, 194 and covered with coarse dark spicules bearing a minute terminal hair; between the anterior marginal tubercles are two small dark elevations, one behind the other, covered with coarse dark spicules; anterior dorso-median tubercles triple, each with small setae internally ; dorso-lateral tubercles each showing two slightly rounded elevations bearing small delicate hairs. Dorsal tubercles represented by a few minute hairs, and on each side of the middle line are three dark puckered macules, forming an irregular row across the dorsum; in front of this row on each side are three similar, if less distinct, macules. Addomen: the arrangement of tubercles and form of the terminal processes are similar to those of the preceding species. Larva. Length 2°6 mm., greatest breadth o'2 mm. (two specimens, apparently not quite mature). Head: length about o'2 mm., greatest breadth 0°14 mm.; well chitinised, conical. Mental plate resembling that of D. fallidihalter, but apparently with five rather large coarse teeth on each side instead of eight. HABITAT: Oblogo; reared from pupae found in collections of water in the bottoms of canoes tied to the bank of the river Densu, also from rotten wood taken from the sides and ends of the same canoes; December, 1919, to March, 1920. Also larvae, pupae and adults in the collection of the Liverpool School of Tropical Medicine obtained from the rotting vegetation at the sides of a water hole at Christiansborg, Accra, March, 1918. Dasyhelea nigricans, sp.n. MEAsUREMENTS. Male. Length of body... 2: onc ane Pec sae .-. I°3 mm. Length of wing... oes ae = 593 eee .. IOmm. Greatest breadth of wing ... oes ; : ag .. O93 mm. Head very dark brown, almost black. Eyes narrowly separated. Clypeus and proboscis dark brown. Palpi brown with dark hairs, third segment the longest, but not so long as the fourth and fifth together and not appreciably swollen, the last segment distinctly longer than the fourth. Awztemnae very dark brown with a plume of black hairs. Segments four to eleven spheroidal, the last four segments more elongate, sub-equal in length, without long spines, the last slightly shorter than the penultimate and without a stylet; segments twelve to fourteen distinctly binodose. 195 Thorax datk brown, almost black, with small yellowish-brown patches at the anterior lateral angles and over the bases of the wings. Scutellum crange brown in the middle, dark brown at the sides, bearing four centro-marginal and two lateral bristles, and one or two median short hairs. Post-scutellum and pleurae dark brown. Wings sparsely clothed with decumbent hairs. The costa terminates about the middle of the wing and proximal to the bifurcation of the fifth vein; the first and third veins form distally a single small cell. Halteres with yellowish-brown knobs and dark brown stems. Legs brown, almost uniformly coloured, the femora and two terminal tarsal segments slightly darker. Claws bifid distally without a sub-basal projection. Addomen: dorsum very dark brown, scantily clothed with black hairs, venter distinctly lighter in colour. Hyrorycium (fig. 8). Ninth segment: tergite long, reaching beyond the ends of the side-pieces, tapering posteriorly and showing Le - 1 me ay TREAD 4 Cc J SISSY \ ; . ‘ 1 : o BSS) ' - i> =" CU SORSu ae Li Shp, eS VS ROE. ; SN . AIC. Fic. 8. D. nigricans, sp.n., male hypopygium (ventral view). X 400 circa. a slight concavity at its posterior margin, clothed dorsally with a few moderately long hairs, finger-like processes well developed, dark, each with a rounded apex bearing a small terminal hair; sternite prolonged posteriorly into a cone-like process. Forceps: side-pieces prolonged distally on their inner sides into sub-conical, thinly chitinised lobes which are clothed with short, stout hairs—the 196 inner margin from the base of each side-piece as far as the origin of the lobes is very heavily chitinised; claspers double, the inner piece more heavily chitinised and shorter than the outer, pubescent, ending in a strongly pointed process which carries a relatively stout hair, and showing a secondary barb on its convex margin at about the apical third, the outer portion less heavily chitinised, pubescent on the basal half or two-thirds and of normal form, with the apex depressed and bearing a few small hairs. Harfes: basal portions rather broad, strongly chitinised, asymmetrical; from the right basal portion there arises a curved, heavily chitinised, blade- like structure which tapers to a point distally, and is prolonged posteriorly as far as the distal end of the main part of the side-piece ; laterally the basal portions are produced into strongly chitinised bars which run diagonally backwards and join the end of the chitinised inner margin of the side-pieces. Aedoeagus: strongly chitinised, broad; the median transverse proximal portion with wing-like prolongations, the lateral portions each composed of a dorsal and ventral plate, the extremities divergent, pointed, resembling a pair of shears. HaBITAT: Accra, Gold Coast. Two males taken in the evening upon the windows of the laboratory, January and February, 1920. Dasyhelea flava, sp. n. MEAsuREMENTS. Female. Male. Length of body (three specimens) aan ... 1:2 mm. 1-3 mm. Length of wing... Se os a .. o8mm. o-9 mm. Greatest breadth of wing ... ace 30 +. 02mm. o-3 mm. Head yellow, the middle of the occiput brown. Eyes separated, narrowly in the female, more widely in the male. Clypeus yellow. Proboscis and palpi brown, the first segment of the palpi very small, the third segment inflated, about one-third longer than the fifth in the female; third segment of palpi in male cylindrical, less inflated than in the female. Awzennae brown with dark hairs; in the female, segments four to ten bear stout sensory spines and are sub-spherical to oval in shape, the length varying from one to one and one-fifth the width, segments eleven to fourteen slightly longer, the last segment tapering to a narrowly rounded apex and almost one and three-quarter times 17 the length of the preceding segment; in the male, segments four to eleven oval to sub-oval in shape (almost hexagonal in transverse section), varying in length from two-thirds to one and a half times the width, last four segments gradually decreasing in length, twelve to fourteen, not binodose but exhibiting the usual sculpturing, with a basal whorl of hairs and shorter hairs distally, the last segment longest, longer than the twelfth, basally very broad and tapering to a pointed apex. Thorax canary yellow with three broad longitudinai brown bands: a median, which runs from the anterior margin to the posterior third of the dorsum and is frequently divided down its middle by a narrow yellow line, and lateral bands extending from the anterior fourth of the dorsum almost to the scutellum. In the male the thoracic pattern is less conspicuous, the yellow markings being contracted so that the general colour appears brown with yellow markings, rather than yellow with brown markings. Scutellum canary-yellow with four centro-marginal and two lateral black bristles, in both sexes; small hairs absent. Post- scutellum dark brown. Pleurae almost entirely canary yellow with small brown markings. Wags hyaline without spots, clothed with decumbent hairs which extend basally well beyond the cross-vein. Halteres canary-yellow in both sexes. Legs brown, rather lighter coloured in the male, the knees infuscated. Claws simple, bifid at the tips in the male. Addomen: dorsum dark brown, venter somewhat tawny, especially towards the apices of the segments. Arising from the centre of the anterior margin of the eighth sternite in the female and extending forwards into the seventh segment is a blunt, chitinous and apparently tubular process (fig. 9, a); the lower laminae appear to arise from the anterior edge of the eighth sternite, but the upper laminae extend backwards internally almost to the edge of the segment. This structure is conspicuous in specimens which have been treated with caustic potash, but is largely obscured by pigment in carbolic preparations; even in dry specimens, however, a small portion of the lower plate can usually be detected near the middle of the anterior margin of the eighth segment. Spermatheca (fig. 9, 6), single, moderately chitinised, sub-spherical to oval (44 by 40) in shape with the beginning of the duct chitinised. Hyrorycium (fig. 10). Ninth segment: tergite broad basally, tapering distally, sparsely clothed with strong hairs, posterior margin 198 not notched and without finger-like lateral processes, sternite relatively narrow, with a conspicuous but not heavily chitinised median pincer-like process projecting backwards ventral to the aedoeagus. Forceps: side-pieces short and stout, broader. apically than basally, rather scantily clothed with moderately long hairs; claspers bifid, highly chitinised, antler-like, the inner branch pointed and bearing two setae a little beyond the middle, the outer branch strongly curved, pointed, with a prominent seta near the base. Harpes: basal portions articulating with the base of the side-pieces, AFC. e Pea WZ, Fic. 9. a—Terminal segments of abdomen of female D. flava, sp.n. (ventral view) showing the chitinous median process arising from the posterior margin of the eighth segment; b—D. flava, spermatheca and median process (side view) ; c—D. fuscipleuris, spermatheca ; d—D. flaviformis, spermatheca ; e—D. similis, spermatheca ; f—D. fusciscutellata, spermatheca ; g—D. pallidibalter, posterior extremity of larva showing the small pseudopod. (a xX 123; bf X 360; g X 490.) strongly chitinised; from the right basal portion arises a lightly chitinised sickle-like process which is very long and _ tapers gradually towards the tip—it is first directed dorsally, but later curves strongly in a ventral direction and distally is covered with delicate spicules. Aedoeagus: a broadly-ending gutter-like structure tapering but slightly from base to apex, the median transverse portion and the lateral portions heavily chitinised. Pupa. Length (two specimens) 1°'7 mm. Respiratory trumpets long, narrow and curved, arising from small tubercles; length about 199 o'2 mm. Distal extremity not infuscated, slightly broadened and without rings, middle portion irregularly ringed, base slightly constricted ; lateral openings of the tracheal trunk occur from base to apex, about six being situated near the tip. Cephalothorax: anterior marginal tubercles large, conical, heavily shagreened, bearing a very short bristle. In the middle line between the anterior marginal tubercles is an antero-posterior row of three Fic. 10. D. flava, sp.n., male hypopygium (ventral view). 400 circa, unarmed somewhat heavily shagreened tubercles, the most anteriorly placed being slightly ventral, the middle one slightly dorsal, to the marginal tubercles. Anterior dorso-median tubercle represented by two or three small hairs; dorso-lateral tubercle some- what prominent, irregularly shaped and bluntly rounded, bearing a moderately long and a short hair; ventro-lateral tubercle bearing a minute hair; ventro-median tubercle represented by a small hair. The ventral surface also bears small, highly chitinised, unarmed tubercles, one on each side of the middle line near the centre of the cephalic region. Abdomen more heavily shagreened than, but otherwise similar to, that of D. pallidihalter. Larva. Adults of this species were reared from larvae obtained from banana fibre which had been isolated in small tubes; the larval 200 pelts were not subsequently found, but as no other species of Dasyhelea was reared from this particular sample of banana fibre in the course of some weeks’ observation, the Dasyhelea larvae found therein have been ascribed to D. flava. Length (three specimens) 2°9 mm.; greatest breadth o°2 mm. Head: length 0°18 mm., greatest breadth 0°12 mm. These larvae are almost indistinguishable from those of D. pallidzhalter, the only apparent difference being in the size and form of the teeth on the mental plate, in D. flava larva the most lateral tooth on each side is smaller, while the others are distinctly larger and coarser than in larvae of D. pallidihalter. HaBITAT: Nsawam and Dodowah, Gold Coast. Reared from rotting vegetable material collected at the bases and roots of banana plants, January to May, 1920. Dasyhelea fuscipleuris, sp. n. MEASUREMENTS. Length of body (one female) ot aA ae fee .. I°2 mm. Length of wing bos aoe oc — “00 Sn .. IOmm, Greatest breadth of wing ... Ee aa ace ae ..» 0-4. 1m. Head brown. Eyes narrowly separate. Proboscis and palpi brown, the fourth and fifth palpal segments short, sub-equal, third segment slightly inflated and rather longer than the fourth or fifth. Antennae brown; segments four to ten inclusive sub-spherical to broadly oval in shape, in length from four-fifths to one and one- quarter times the greatest width, segments eleven to fourteen oval and in length about one and a quarter times the greatest width, the fifteenth slightly longer and about twice as long as wide. Thorax yellow, becoming yellowish-brown posteriorly in front of the scutellum, with broad brown or blackish-brown median and lateral bands; the median band, which has a shining appearance in certain lights, is of almost uniform width and extends from the anterior margin to about the middle of the thorax, the lateral bands are broad and rounded anteriorly, gradually narrowing posteriorly, and extend from the anterior fourth of the thorax almost to the scutellum ; each lateral band near the middle of its outer border gives off a narrow dark stripe which is directed forwards and ends a short distance behind the antero-lateral angle. Scutellum entirely pale 201 yellowish-brown with ‘three or four centro-marginal and two lateral bristles; short hairs absent. Post-scutellum dark brown. Pleurae almost entirely yellowish-brown with a few small darker brown markings. Wezxgs hyaline, sparsely clothed with decumbent hairs which scarcely extend basally beyond the cross-vein and in the anterior apical portion are almost confined to the margin and to the fold above the upper branch of the fourth vein. The costa extends slightly beyond the middle of the wing and distinctly beyond the bifurcation of the fifth vein; the singie cell between the first and third veins distinct, rather longer than broad. MHalteres yellow-brown. Legs: distinctly light brown with the extreme bases of all the tibiae and the apices of the hind tibiae very dark brown; the fifth tarsal segments of all the legs are somewhat infuscated. Claws simple. Abdomen: dorsum dark brown, with the hind margins of the segments narrowly but distinctly yellowish; venter of a rather pale brown colour. Spermatheca (fig. 9, c) single, oval to pyriform (average about 614 by 42), highly chitinised, the commencement of the duct chitinised for a short distance (about 8). HABITAT: Accra, Gold Coast. Two females taken cn the arm, one in a bungalow near a light at 9 p.m., April 18th, 1920, and the other in the laboratory in May, 1920. Numerous specimens (Q 9) bred later. A yellowish-brown midge somewhat resembling D. flava in appearance, though less brightly coloured. Dasyhelea flaviformis, sp.n. MEAsuREMENTS. Female. Male. Length of body... Ere oa er .. I-Omm. I-o mm. Length of wing... fe et Seam .. o-6mm. o-7 mm. Greatest breadth of wing ... ee nat .. O2mm. 0-2 mm, Head brown with lighter brown borders round the eyes and occiput. Eyes narrowly separate. Proboscis and palpi brown, third segment of the palpi not swollen, longer than the fourth and about as long as the fifth segment. Antennae brown with dark brown hairs; segments four to ten in the female with curved spines, sub-spherical, as broad as long, segments eleven to fourteen rather longer than broad, the terminal segment the longest, being two and a half times as long as broad and almost twice as long as the 202 preceding segment: the last four segments of the male elongate, segments twelve to fourteen binodose. Thorax tawny-yellow with broad, longitudinal, brown bands arranged in much the same pattern as those of D. flava and of D. fuscipleuris: a broad median band, divided by a narrow, median tawny stripe extends almost from the anterior margin to the posterior third of the dorsum, and laterai bands, one on either side, extend from the anterior third of the dorsum almost to the scutellum. Scutellum tawny-brown with slightly darker brown sides, carrying three central and two lateral bristles in both sexes; short hairs absent. Post-scutellum dark brown. Pleurae tawny with two or three dark brown markings. Wings hyaline; decumbent hairs rather more scanty than in D. flava, extending basally between the fourth and fifth longitudinal veins to about the point of separation of the first and third longitudinal veins, the anal cell bare and the anterior apical region rather more sparsely clad. Halteres with brown stems and cream-coloured knobs. Legs Fic. 11. D. flaviformis, sp.n., male hypopygium (ventral view). x 400 circa. almost uniformly light brown, with dark knee spots. Claws simple and equal. A&domen dark brown; in the female there are semi- lunar tawny markings on either side of the middle line on the dorsum of the fifth, sixth and seventh segments, and the apex of the abdomen is pale; in the male entirely dark, except for small paler lateral spots on the first and second segments. Spermatheca (fig. 9, d) pyriform (length 43, breadth 35), moderately highly chitinised, the commencement of the duct chitinised. Hyprorycium (fig. 11) resembling that of D. flava. Ninth segment: tergite broad basally, narrowing slightly towards the 203 apical margin which is not notched, finger-like lateral processes rudimentary but bearing a single rather long stout hair; sternite with the median, pincer-like extension well developed. Forceps: side-pieces relatively longer than in D. flava, claspers more slender and less curved, and bearing a rudimentary, conical, second branch on their inner sides basally, the basal portion only pubescent. Harpes similar to those of D. flava but relatively larger and stouter ; the sickle-like structure arising from the right basal process is very long and strongly curved, and apically is devoid of spicules. Aedoeagus differs from that of D. flava in that the distal extremity gives off two curved chitinous processes, one on either side of the middle line, resembling a pair of forceps. Pupa. Length about 1°5 mm.; somewhat infuscated at the anterior end, integument shagreened. Resfiratory trumpets (fig. 3,5): length about 015 mm.; breadth about 15m. They are infuscated apically, and are long, narrow and curved, arising from slight papillae; the main tracheal trunk begins to give off lateral branches just above its base, the first four widely separate but those near the tip more closely situated, together forming a series of nine or ten branches. The trumpets are irregularly ringed in the middle and show no definite tubercles. Cephalothorax infuscated anteriorly; operculum dark, highly chitinised and _ coarsely shagreened, with two low and rounded, but relatively large, elevations in the middle line. Anterior marginal tubercle well- developed, conical, bearing a very small seta; dorso-lateral tubercles well-developed, bearing several minute hairs; anterior dorso-median tubercle almost indistinguishable, being represented by a few minute hairs; ventral tubercles undeveloped, represented by minute hairs. Dorsum of thoracic region infuscated, without tubercles but with numerous macules. Addomen exhibiting no peculiarities, of the usual pattern but rather poorly chitinised. HABITAT: Oblogo, Gold Coast. Reared from rotten wood taken from a canoe in the river Densu, May, 1920. The pelt of a larva which had been isolated in a tube with a piece of the rotten wood in which it was found and from which a pupal pelt and an adult of this species were obtained, was not recovered. A small brown midge resembling in markings D. flava, but with the lighter coloured areas a dull tawny-yellow in place of brilliant canary-yellow. 204. Dasyhelea fusca, sp. n. MEeasurEMENTS. Female. Male. Length of body... He ne a seouse ame 1-6 mm, Length of wing 335 S05 ae sce foe Usha) 1-3 mm. Greatest breadth of wing ... 5 o-4 mm. o-3 mm. Head brown with brown hairs. Eyes broadly contiguous in the female, narrowly separate in the male. Proboscis pale brown. Palpi brown, third segment slightly swollen, distinctly shorter than the fourth and fifth segments together. Azz¢enxnae in the female brown, torus much darker than the segments of the flagellum, with dark hairs and large, curved spines (in addition to short blunt spines) on all the segments; segments four to eleven sub-spherical to oval, constricted apically, the last segment elongate, at least one and a half times as long as the penultimate, with a distinct stylet. In the male, strongly plumose, the whorl hairs dark brown; segments three to eleven spheroidal, segments twelve to fourteen elongate, strongly binodose; last segment stout and cylindrical, with a conspicuous stylet. Thorax dark brown with paler brown to yellowish areas at the anterior lateral angles and bases of the wings (in fresh specimens the dorsum is greyish-pruinose), sparsely clothed with dark brown hairs. Scutellum pale yellowish-brown with five or six centro-marginal and two lateral bristles, and about six (four to seven) short median hairs. Post-scutellum and pleurae dark brown. Wezxgs almost unicolourous but slightly infuscated at the junction of the first and third veins and costa; surface rather thickly clothed with long hairs, which extend basally beyond the cross-vein. First and third veins fused, extending for a considerable distance parallel to the costa and joining the latter well beyond the middle of the anterior border and the bifurcation of the fifth vein. Halteres with cream-coloured knobs. Legs light brown, the knees slightly darker, the apices of the tarsal segments infuscated, fore and hind femora each with a dark spot -above the apex. Claws simple, bifid distally in the male. Addomen dark brown with brown hairs, which are most numerous at the sides and on the distal segments. Spermatheca more or less pyriform (length 61m, breadth 45) in shape and feebly chitinised. Hyporycium (fig. 12). Ninth segment: tergite with moderately long hairs dorsally, broad, gradually tapering towards the posterior 205 margin, the finger-like processes well developed, dark, each with a stout hair at the apex and a group of five or six shorter hairs at the base ; sternite relatively broad, without a central posterior projection. Forceps: side-pieces short and broad, clothed with short, stout hairs; claspers pubescent, rather longer than the side-pieces, the tips slightly rounded and bearing ~a few minute hairs. Harfes: basal portions somewhat asymmetrical, large, strongly chitinised, curving forwards towards their junction near the middle line; distal portion Fic. 12, D. fusca, sp.n., male hypopygium (ventral view). x 400 circa, of the right harpe in the form of a stout, strongly chitinised, blade- like structure with a pointed, slightly re-curved apex, extending posteriorly almost as far as the posterior margin of the ninth tergite. Aedoeagus large, projecting posteriorly to about the level of the middle of the side-pieces, broad basally, narrowing slightly apically as shown in fig. -12. Pupa. Length 2°3 mm.; relatively large and well chitinised, with the cephalo-thoracic portion slightly infuscated and with 206 characteristic, loofah-like, trumpets. Integument shagreened, especially over the middle of the anterior part of the cephalothorax and on the last abdominal segment. Respiratory trumpets (fig. 3, d): length about 0°3 mm., broad and flat, arising from small tubercles; the basal third cylindrical, dark coloured, annulated, the apical two- thirds expanded and flattened, with very numerous tracheal branches arranged somewhat like the leaflets of a long pinnate leaf which has been doubled upon itself so that the fold is at the distal end and the base and tip at the proximal end—the branches are apparently double processes arising from the main tracheal trunk. Cefhalo- thorax: anterior marginal tubercles prominent, conical, divergent, covered with coarse dark granulations and ending each in a double- pointed process bearing a delicate hair; anterior dorso-median tubercles somewhat irregularly shaped and bearing small and delicate setae on their inner sides; dorso-lateral tubercles highly granular, bearing one small and one minute seta. On each side of the anterior marginal tubercles is a dark, conical, coarsely granulated nodule, and immediately dorsal to this a smaller and flatter granulated nodule. Dorsal tubercles apparently represented by socket-like marks (which may be surrounded by a dark zone and may show centrally minute points resembling small spines) and by small dark puckered macules—there are about eighteen of the latter, of which fourteen (seven on each side of the middle line) are arranged in an irregular transverse row extending across the dorsum a short distance posterior to the trumpets, and the others are situated anteriorly two on each side of the middle line. Large dark areas, which appear to be slightly raised, also occur over the bases of the wing sheaths and on that part of the cephalothorax immediately posterior to them, and other macules may be distinguisned on the posterior portion of the dorsum. Aédomen: tubercles poorly developed ; on both surfaces of each segment, but more conspicuous on the dorsum, are three rounded and darkened macules arranged in the form of an equilateral triangle with its apex posteriorly directed. HABITAT: Oblogo, near Accra. Pupae and pupal pelts were found in stagnant water contained in canoes which were tied to the bank of the river Densu; and pupae in the rotten wood forming the sides and ends of these canoes. Larvae were not obtained. 207 Dasyhelea nigrofusca, sp. n. MeasurEMENTS. Male. Length of body _... ee a ie ows Soe eee bay nial Length of wing... ae ae ae Ste Re ses, 7 LO In. Greatest breadth of wing ... se ace Sec te ... O'3 mm. Head dark brown, almost black. Eyes broadly contiguous dorsally. Proboscis and palpi dark brown, the third segment of the palp distinctly swollen and rather longer than the fourth or fifth, which are sub-equal in length. Aw‘ennae dark brown with almost black hairs ; segments four to eleven more or less hexagonal, in length from three-fourths to rather more than once the width, segments twelve to fourteen elongate, binodose, decreasing in length from about three times to two and a half times the width, terminal segment broad, rather shorter than the twelfth segment and with a small apical stylet. Zhovax dark greyish-pruinose with sepia markings, the lateral margins narrowly yellowish; scantily clothed with dark hairs. Scutellum dark brown in the middle, yellow laterally, with a row of eight sub-marginal bristles and about half a dozen median short hairs. Post-scutellum dark brown. Pleurae yellow with dark brown markings. Wemgs hyaline, well clothed with decumbent hairs, First and third veins apparently completely fused, the bifurcation of the fifth vein lying immediately below the apex of the costa. Halteres brown with paler coloured but slightly infuscated knobs. Legs conspicuously banded, femora dark, slightly paler basally and with a distinct pale band before the apex, knee joints dark, tibiae with dark bands at the base, middle and apex, tarsal segments paler, with indications of dark bands at the apices; clothed with dark hairs. Claws simple and equal, bifid at the tips. Abdomen sepia-coloured, clothed with dark brown hairs; venter yellowish. Hyropycium (fig. 13). Ninth segment: tergite scarcely tapering, the posterior margin straight, the finger-like processes well developed, each bearing a single hair and with a prolongation of the tergite connected with them dorsally; sternite without a central posterior extension. Forceps: side-pieces of the usual form; claspers as long as the side-pieces, the basal halves pubescent and carrying two short hairs near the middle and three or four short hairs near the apex. Hares: basal portions strongly 208 chitinised and bent in an anterior direction, that of the right side being shorter and more abruptly bent and articulating with a double curved, blade-like distal process; the latter is very long and reaches posteriorly as far as the apices of the side-pieces, it is broader in the middle than at either extremity, and is serrated at the apex. Aedoeagus broad basally, narrowing somewhat apically, the distal portion consisting of two highly chitinised hook-like rods on either side connected by a ventral wall of thin chitin; these lateral rods are not on the same plane, and the ventral pair are longer, less sharply bent medially, and notched at the ends. Fie. 13. D. nigrofusca, sp.n., male hypopygium (ventral view). X 400 circa. Pupa. Length 26 mm.; shagreened but not highly chitinised. Respiratory trumpets (fig. 3, ¢): length about o'11 mm., short, broad and constricted at the base, arising from small tubercles and with the basal half covered with minute squamose spines; lateral processes (numbering about twenty) of the tracheal trunk given off at regular intervals—at first in a single, later in a double row, extending from a short distance above the base to the apex. Cephalothorax: spines and tubercles poorly developed ; operculum 209 wanting. Anterior marginal tubercle small, bearing a small bristle ; dorso-lateral tubercle bearing two small hairs; anterior dorso-median tubercle very small with two hairs; ventro-median tubercle minute, bearing a small hair; ventro-lateral tubercle represented by two or three small hairs. Dorsum of the thoracic portion without tubercles or spines. Addomen: integument with transverse cellular sculpturing and minute spicules basally and laterally ; these spicules are most numerous and most highly developed on the last segment and its terminal processes. The segments are furnished with the usual backwardly projecting shelf-like tubercles, those forming the ventro-lateral series being the most prominent. HABITAT: Dodowah, Gold Coast (north-east of Accra), reared from materials collected from a rot-hole in a mango tree, March 7th, 1920. The larva was not identified. Dasyhelea fusciformis, sp.n. MeasurEMENTS. Female. Length of body... Pe GES ae SEE eas diego Length of wing... “ee aS Ae “AE a con Lode Inter. Greatest breadth of wing ... Bes se ie es .-. 0°73 mm. Head brown with brown hairs. Eyes very narrowly separate, almost contiguous. Clypeus, proboscis and palpi brown; all segments of the latter somewhat swollen, the third cylindrical, longer than the fourth and fifth segments together, the fifth much shorter than the fourth. Ax/ennae brown with brown hairs and long curved spines on all the segments; segments of the flagellum sub-spherical to oval in shape, each with an apical constriction, the last. segment elongate with a distinct stylet (fig. 2,@). Thorax dark brown with dark hairs, the anterior-lateral angles and areas at the bases of the wings paler brown. Scutellum light brown with eight centro-marginal - and two lateral bristles, and three short hairs. Post-scutellum and pleurae dark brown. Wzngs: decumbent hairs extending to the base of the wing between the fourth and fifth veins; costa reaching the middle of the anterior border and terminating beyond the bifurcation of the fifth vein, the first and third veins distally forming a small cell. Halteres with cream-coloured knobs and dark brown stems. Legs uniformly brown, the claws simple. Ad&domen dark brown, lighter coloured distally, with dark brown _ hairs. 210 Spermatheca sub-spherical (length 53“, width 48), moderately chitinised, the duct wide at its origin, tapering gradually and chitinised for a very considerable portion (50) of its length. Pupa. Length 2 mm., moderately well chitinised, the integu- ment shagreened and the last abdominal segment with small spicules. Respiratory trumpets: length about 0°17 mm., breadth 0°03 mm., rather short, broad and almost straight, with a slight basal constriction and with the surface covered with minute, imbricated, flattened spines. The tracheal trunk gives off three or four lateral branches at regular intervals in the basal three-fourths of the trumpet, and a series of about a dozen close together in the apical fourth. Cephalothorax: anterior marginal tubercles wanting —the operculum missing from the single pelt examined; anterior dorso-median tubercle poorly developed, bearing a single small hair ; dorso-lateral tubercle small, carrying two short hairs; ventro-lateral tubercle indistinguishable; ventro-median tubercle represented by two small hairs. Integument of the dorsum of the thoracic portion with puckered macules, but without tubercles. Abdomen with tubercles of the usual type and arrangement. The pupal stage is apparently short, as the adult insect appeared on the fourth day after isolating the larva. The pupal pelt was found with the cephalothorax protruding from a piece of rotten wood. Larva. A single larva of this species was collected and isolated in a small tube; it was rather large, with well-formed hooks at the posterior end of the abdomen, Its movements were slow and deliberate and it was observed to crawl up the sides of the glass vessel in which it was contained, but afterwards it quickly buried itself in the rotten wood and was not seen again. After the insect had hatched the pupal pelt was secured, but the only portion of the larval pelt found was the head; this was dark-coloured and heavily chitinised, and measured 0°3 mm. in length and o'2 mm. in greatest width. HABITAT: Oblogo, Gold Coast. Reared from materials collected from a rot-hole in a tree (Cynometra sp., probably C. megalophylla, Harms.), May, 1920. D. fusciformis is a brown midge closely resembling D, fusca, but smaller in size and with the colouring of the scutellum and the lighter 21f patches on the thorax. pale brown rather than yellow. The spermathecae and pupae of these two species are quite distinct. -_ . The species of Dasyhelea described above may be separated as follows :— FEMALES Antennal segments sub-spherical or oval, not constricted apically, ‘the last segment not produced into.a stylet ; scutellum with at -Most four centro-marginal bristles Antennal segments constricted apically, the last with a distinct stylet ; scutellum with at least five centro-marginal bristles ... Species mainly dark brown in colour ... Species yellowish with dark brown markings ... Scutellum dark brown, at most with the central portion yellowish... Scutellum entirely yellowish Scutellum yellowish in the middle ; halteres white, padlidihalter, sp. n. (p. Scutellum entirely dark brown ; halteres yellowish ... Spermatheca reniform; claws with a distinct sub-central pro- - jection ar fe aia ave ... fusciscutellata, sp. n. J » Sp Spermatheca spherical ; claws without a distinct sub-central pro- jection es Hceve) We Re cs w= similis, sp. n. Scutellum bright yellow ; halteres white ..._ Jutcoscutellata, sp. n. Scutellum yellowish-brown ; halteres yellowish-brown as sua «+. INconspicuosa, sp. Species bright yellow ; the eighth abdominal segment with a ventral backwardly directed chitinous process... .» flava, sp. n. Species dull yellow ; the eighth abdominal segment without such a process So RR aa hse Larger species (wing length 1-o mm.) ; halteres yellowish-brown ; hind margins of abdominal segments narrowly yellowish ... fusctpleuris, sp. n. Smaller species (wing length 0-7 mm.) ; halteres cream-coloured ; semilunar tawny markings on dorsum of fifth, sixth, and seventh abdominal segments oe an + Haviformis, §p. n. Spermatheca pyriform, feebly chitinised ; scutellum with five or six centro-marginal bristles... ar er .. fusca, sp. n. Spermatheca spherical, moderately chitinised, with a very long posterior extension ;. scutellum with eight centro-marginal bristles ee occ ee rE «. fusciformis, sp. n. (p. (p. - (p. (p. (p. (p. (P. 212 Mates (separation on hypopygial characters) 1. Claspers single 50 Claspers double ... fos so 2. Ninth sternite with a central posterior extension > Kw Nn vd Ninth sternite without a central posterior extension see see 3. Posterior extension of ninth sternite large, ob-ovate ane “6 so) Se w+ eee pallidihalter, sp. n. (p. 184) Posterior extension of ninth sternite smaller, somewhat rectangular xe sas bes ee bas ... fusciscutellata, sp. n. (p. 187) Posterior extension of ninth sternite cone-like — imcomspicuosa, sp. n. (p. 191) 4. Distal portions of harpes fused, forming a single median process ... ees ace Bec nae ae occ .-. similis, sp. n. (p. 189) Distal portion of one harpe only developed ... sau Uiseaain pees a) 5. Aedoeagus relatively short, reaching about one-third the length of the side pieces ; distal fourth of the harpal process attenuated, the apex serrated... ee eee ..- nigrofusca, sp. n. (p. 207) Aedoeagus long, reaching beyond the middle of the side pieces ; harpal process not attenuated distally, the apex pointed — oe Ges aoe see Aas co w. fusca, sp. n. (p. 204) 6. Inner portion of clasper small, conical w- _ flaviformis, sp. n. (p. 201) Both portions of clasper large and well developed ... “ oe 7 7. Central posterior extension of the ninth sternite cone-like ... eee S55 es =e =e os = nigricans, sp. n. (p. 194) Central posterior extension of the ninth sternite pincer-like ae 55 $e: oe sor ... flava, sp. n. (p. 196) REFERENCES Carter, H. F. (1919). New West African Ceratopogoninae. Ann. Trop. Med. & Parasit., Vol. XII, pp. 289-302. Carter, H. F., Incram, A., and Macrtr, J. W. S. (1920). Observations on the Ceratopogonine Midges of the Gold Coast with descriptions of new species. Part IL: Ibid., Vol, XIV, pp- 211-274. Part III: Jbid., pp. 309-331. Kuerrer, J. J. (1917). Chironomides d’Amérique conservés au Musée National Hongrois de Budapest. Ann. Mus. Nat. Hung., Vol. XV, pp. 292-364. (1919). Chironomides d’Europe conservés au Musée National Hongrois de Budapest. Ibid., Vol. XVI, pp. 1-160. (1919). Observations sur les Chironomides (Dipt.) décrits par J. R. Mailoch. Bull. Soc. Ent. de France, No. to (May), pp. 191-194. Mattocs, J. R. (1915). Some additional records of Chironomidae from Illinois and notes on other Illinois Diptera. Bull. Ill. State Lab. Nat. Hist., Vol. XI, pp. 305-363. 213 THE PREVALENCE AND CHARACTER OF TUBERCULOSIS IN HONGKONG BY HENRY HAROLD SCOTT GOVERNMENT BACTERIOLOGIST, HONGKONG (Received for publication 1 July, 1921) I. GENERAL CONSIDERATIONS The statement has been made that not only is tuberculosis a common disease in the tropics, but that it appears to be increasing in its ravages in the East. Seeing that the treatment of tuber- culosis, both prophylactic and curative, consists largely in fresh air and sunshine, it seems strange that this disease should be rife in tropical countries where sunshine is so prevalent and living in the open air is the rule. A very brief experience as medical officer in charge of the mortuary sufficed to show that tuberculosis was a frequent cause of death in Hongkong, and I deemed it an investigation well worth undertaking to determine the varieties of the disease as met with here, the portals of entry, the mode of spread, and, if possible, to determine the primal cause or causes responsible for the condition and those aiding its dissemination, with a view to elucidating measures for its prevention. I have excluded those cases which showed merely signs. of old-standing tuberculosis, for previous investigations have proved that after middle life nearly all bodies yield evidence of some healed focus of the disease. A particularly noticeable feature in my series is the large percentage of children of tender age who die from tubercn- losis. Another point worthy of note is the fact that whereas at home intestinal tuberculosis as a primary condition is fairly common in children and is ascribed to the drinking of infected milk, in tropical countries primary intestinal tuberculosis is met with although milk 214 is not taken as a regular food; this holds good here also, though it is only right to add that primary intestinal infection appears to be comparatively rare in Hongkong. Before proceeding to speak in detail of the cases dealt with in this investigation, it will not be out of place to make a few general remarks on the disease. As is well known, the bacillus may persist for a long time in dust, in dried sputum, in urine, and so forth. When one sees how large a number of patients exhibit signs of tuberculosis after death, several of whom showed no marked signs during life, and when, as we know, the bacilli are shed broadcast not only in the sputum but in the fine spray of coughing and sneezing consumptives, the opportunities for the organism to gain entrance and to propagate are many, Thus, the bacillus has been found in the sweepings of rooms (Manfredi at Naples, Marpmann at Leipzig), on fruit exposed for sale (Schnirer), in the wards of hospitals (Strauss, Le Noir), in books, in the bodies of flies (Nuttall, André), on clothes (Josephson, Nétel, Chaussé), as quoted by Etienne Burnet: ‘Ou trouve comme une quintessence des poussiéres atmospheriques dans ces poussiéres fines que l’on pompe par le vide (vacuum cleaner); avec elles s’est déposé ce qu'il y a de plus léger dans ce que soulévent nos semelles et l’intense circula- tion de nos rues, et ce que crache une population malheureusement trop insouciante.’ As stated by Delépine, the difficulties of elucidating the causes of an infectious disease depend on the fact that we must take account of the various ways in which the predisposing and determining factors have combined in their action, and the difficulties are increased when we attempt to assign to each its appropriate share. He groups these factors under the following heads: (1) Distribution and habits of the pathogenic organism. (2) Conditions influencing its number. (3) The pathogenicity and toxicity of it, or, in other words, the conditions influencing its virulence. (4) The oppor- tunities which the organism may have of gaining access to one or more channels of infection. (5) The conditions influencing the resistance which any possible host may be capable of opposing to the attacks of the organism. (6) The frequency and degree of completeness of recovery from occult or manifest infection. This comes into play more with regard to the question of adult tuber- 215 culosis, whereas the present investigation is concerned with the disease as found in children. Of these factors those of special importance are the number and virulence of the organism on the one side and the degree of resistance which the subject attacked may be able to oppose on the other. There appears to be no doubt that the resistance set up, if not adequate for the destruction of the organism, even contributes indirectly to an increase in the virulence of the latter. Thus, an organism, ¢.g., the Bacillus tuberculosis of reduced virulence in milk, after inoculation was found to gain in virulence according as the distance from the site of inoculation increased. The resistance offered to the invading organism being insufficient to destroy it, trained it, as it were, to develop more inimical powers of attack, so that, as the bacilli overcame. successively the barriers placed in their path, they increased not only in virulence but in number also, just as the Virus five of rabies was obtained by Pasteur by means of passage of the street virus through a series of animals. This process has been designated a ‘reinforcement by relays.’ The overcoming of this resistance by an organism of feeble aggressive powers of course takes time whether the delay is caused by the small number of organisms, or by their being of reduced virulence, or by the greater degree of resistance set up leading to a ‘relatively reduced virulence.’ One or other of these, acting singly or in combination, may be adduced to account for latency. Where cases are met with in which death occurs at such a tender age as 22 days (No. 128),* 24 days (No. 214), 29 days (No. 180), and 7 weeks (Nos. 63 and 114), one is naturally led to enquire into the question of congenital and -hereditary tuberculosis. The possible modes of congenital infection are, of course, either by the ovum or spermatozoon, or via the placenta from a tuberculous mother. As regards the first of these, as Adami states, ‘ the microbe of an infectious disease cannot be a constituent of the biophore. At most it can be an accidental inclusion in the surrounding non- heritable matter of the cell.’ The human ovum, being practically * The serial numbers refer to those contained in a table showing the post-mortem findings of the 300 cases of tuberculosis under consideration. The Editors regret that it is not possible to reproduce this table, ‘ 216 free from yolk and not being phagocytic, cannot take up the bacilli, while it is still more improbable that the minute spermatozoon could carry them. Repeated observations go to show that the semen of a phthisical patient (Rohlff, Gaertner) does not contain as many as ten bacilli, and as the average human seminal ejaculation is said to contain over 200 million spermatozoa, the chances that a single spermatozoon which fertilises the ovum should carry a bacillus are about one to twenty million. For practical purposes this mode of infection may be regarded as impossible. The second method, via the placenta of a tuberculous mother (or perhaps through the walls of the foetal sac, or passing to the foetus before the sac forms), has been shown to be possible; in fact, intra-uterine acquirement of disease as distinct from inherited disease is proved. Such trans- mission of the tubercle bacillus from parent to child im wtere, is however, undoubtedly very rare. Thus Schluster, who examined into the records of the reported cases, as mentioned by Latham, was able to collect only twelve showing clinical evidence of tuberculosis at birth. Furthermore, in the majority of instances, the organs of foetuses born of tuberculous mothers yield only negative results when inoculated into guinea-pigs. The theory of diathesis—the question as to whether the children of tuberculous parents suffer from a greater liability to tuberculosis from enhanced susceptibility resulting from ‘ paratuberculous lesions ” due to toxic action on the germ cells—the hereditary transmission of constitutional predisposition to tuberculosis, is being relegated more and more to the realms of improbability as modern knowledge, with its insistence on accuracy of detail, demands facts and proofs in place of nebulous hypotheses. The dominating factor in the incidence of tuberculosis is the opportunity for infection; in other words, the greater the oppor- tunity for infection the greater is the incidence and the mortality from this disease. Can we argue that the reverse of this also holds good, namely, that a high incidence and mortality mean greater opportunities for infection? Probably we can, as will be shown later when the local conditions are mentioned. It would appear to be established that under certain circum- stances, no matter what the ‘ diathesis’ may be, infection can always be effective. To prove that some children inherit a predisposition 217 to tuberculosis, the incidence of the disease amongst theoffspring of tuberculous parents and amongst equal numbers of non-tuberculous parents would have to be compared, and, furthermore, it would be essential that both should be subjected to the same environment from birth, as McFadyean has pointed out. Even then we must bear in mind the fact that the former will have been exposed to infection during the intra-uterine period, so that the conditions for comparison will not be strictly impartial. To compare, as has been’ done, the death-rate from tuberculosis amongst members of the two categories—the children of healthy parents on the one hand and those of tuberculous parents on the other—is fraught with fallacy, as the after conditions are so diverse. To make them more equable it would be necessary for the children of healthy parents to be born in infected houses or be placed there at birth, clearly an impossible condition except by mere accident. The children of tuberculous parents are thus handicapped in several ways, each of which plays a part in rendering fallacious the comparison of their death-rate with that among children of healthy parents. The chief of these disturbing factors would be: (1) Intra- uterine exposure. (2) Birth in infected surroundings, and, therefore, greater liability to infection while young. (3) The dose of infecting material is likely to be large. (4) This dose is probably frequently repeated. That the intra-uterine period may be dangerous to the child from the point of view of tuberculosis has been definitely proved, though the cases on record are very exceptional. Thus Schmorl and Birch-Hirschfeld, and Schmorl and Kockel, have described cases in which the bacilli were found in the placentae and the foetuses of tuberculous mothers. More recently Friedmann reported the finding of two cases of tuberculous infection by the placental route. So much for the question of hereditary transmission of the Bacillus tuberculosis. As regards environment no one now disputes the im:portance of this factor in producing the disease, but though comparison is difficult in human beings owing to the variety of disturbing factors, as has already been stated, in animals we can better arrange conditions to suit our purpose. The experiments of Trudeau may be referred to in this connection. He inoculated 218 rabbits With tuberculosis and allowed some to run wild, whereas others were kept in a damp, dark place. Most of the former recovered, while the latter rapidly succumbed. In cattle an analogous condition is found. As McFadyean states: ‘ All breeds and strains of cattle are susceptible to tuber- culosis, and when the environment is the same the incidence of the disease is the same in all breeds and strains.’ He found that the conditions under which the cattle are bred and reared constituted the most important contributory factor according to the opportunity supplied by the environment for the transmission of the bacilli. Regardless of breed as a separate factor, the proportion of cases of tuberculosis furnished by any breed is high or low according as the cattle are or are not in close association with other diseased animals. Calmette’s experiments lend additional support to the same fact. He found that cattle which had been once infected nearly always recovered if they were carefully isolated, but if kept in prolonged contact with tuberculous animals they themselves became actively tuberculous. So in man no race is exempt; in other words, there is a racial susceptibility to tuberculous infection, but there is not sufficient evidence at present to enable one to say that any special predisposition is handed on by tuberculous parents to their offspring. We may summarise the matter by saying, firstly, that hereditary transmission of the bacillus is so rare that for practical purposes it may be declared negligible; secondly, that the incidence of tuber- culosis depends on two main factors, namely, the dose received and the virulence of the strain inoculated; in other words, the degree of exposure to infection and the resistance which the inoculated subject is able to put forward, this, in turn, being dependent largely on his environment; thirdly, that there is not sufficient evidence, at present at all events, to afford support to the theory that there is such a thing as inherited predisposition to tuberculosis. Before passing on to a more detailed description of some of the 300 cases of death from tuberculosis, the morbid anatomy of which forms the basis of these studies, it will be advantageous to say a few words on the extent to which the foregoing points are exemplified in the conditions prevailing in Hongkong. 219 Generally speaking, the prevalence of the disease is closely connected with social.and economic conditions—overcrowding and slums, poverty, insanitation, and squalor. The ingestion of tuberculous milk, which is supposed to play a large part in the production of tuberculosis, especially intestinal tuberculosis, in» children in England, has no influence here, for the Chinese children do not drink milk, nor does it arise here from the use of tuberculous meat. The problems of tuberculosis in Hongkong are really social problems, and are, therefore, intimately connected with those of public health. The main causes of the prevalence of the scourge are the predisposing ones of overcrowding of the poor and the fact that the rooms inhabited by them are dark and the sunlight rarely enters them. They are still further darkened by gratings and shutters. With the first of these there is little if anything to be done; the population is great and the space for their accommodation relatively small; not only is floor space inadequate, but window space is less than it should be, and the windows are often closed and made of opaque or coloured glass, so that the penetration of light is reduced toa minimum. This latter question of the darkening of the rooms can only be improved by education. In an interesting paper on “Sanitary Progress in Hongkong,” Dr. W. W. Pearse, the Medical Officer of Health, states with reference to the housing of the poorer Chinese that, the area being limited and the Chinese population large and constantly increasing, building sites have become very expensive so that the streets inhabited by this class of person are narrow, and the houses fronting on them high; in some cases the height is five times the width of the street. ‘It is usual,’ writes Dr. Pearse, ‘to speak of the Chinese houses as being of one or more storeys, but as each storey was generally let separately, and as the occupiers of each storey used it as they would have used their single storey house in their native village in China, in effect these Chinese houses in Victoria were piled one on another, three, four and five high! “Another factor has contributed curiously to produce the ill- designed Chinese house of Hongkong. The floors of these houses are supported by China fir poles. A pole of more than fifteen feet 220 long of sufficient strength for a floor joist is not readily procurable. This has limited the width of storeys to fifteen feet. “In order to provide as much accommodation per floor as possible, z.e., to make building pay, the area of a floor has been obtained by increasing its depth out of all proportion to its width. ‘Hence a Chinese house in Hongkong has been a veritable tunnel fifteen feet wide and forty, fifty and even sixty feet deep. ‘Excepting corner houses only, windows to light and ventilate these tunnels were possible only at the front and in front of these were verandahs. ‘Windows at the rear of houses were not at first considered necessary, and many houses were built back to back, with no yard or ventilating shaft between them. On each floor the rear portion was cut off by a partition wall to form a kitchen. Such kitchens were small, generally under 150 square feet in area, very dark, and often with no means of ventilation other than the door communt- cating with the main portion of the floor. These kitchens were drained by vertical earthenware waste pipes which, in the case of back to back houses, were carried down through the building to discharge over a gully trap in fhe kitchen on the ground floor.’ From time to time laws have been passed to improve the housing conditions by providing open spaces of a defined minimum area at the rear for ventilation. Later, all new houses were required to have larger back yards, and also scavenging lanes. Further, regular systematic cleansing of the Chinese houses was instituted (in 1903) and certain repairs effected. These measures were really directed towards the eradication or limitation of plague, and were aimed at affording protection from infestation with rats. The method of cleansing the houses is carried out in the following manner four times a year: the furniture 1s turned out, the bed-boards and such like are dipped in 1 per cent. kerosene in water, in order to get rid of vermin; the floors also are sprayed with the same liquid. All rubbish and dirt generally are removed, and an inspector makes a careful examination to see if any floors need repair, if any gratings are missing, and so on. The accompanying diagram, kindly drawn for me by Dr. W. J. Woodman, late Acting Medical Officer of Health for Hongkong, and now M.O.H., Kewleen, shows the arrangement and mode of 221 occupation of a typical ‘floor’ of a modern Chinese house in these districts. The foregoing constitute the main predisposing causes, but the direct cause of the prevalence of tuberculosis in Hongkong is, facile princeps, the expectoration habit. Nearly all writers on tuberculosis in the tropics note this peculiar trait. In the houses of the poor a person will expectorate anywhere on the floor, or, if he is ill in bed, on the bed clothes. Owing to the unavoidable over- crowding in the poorer districts the children play about on the floor, putting everything into their mouths, breathing a vitiated UE ECONE SIE ECR RO EERE. 11 Egan aan Sepa aerate > = B BT} 1h. G ’ g ca i Qrca nai’ less 3} ae : 2} é Ce ’ a} a § Kanbuyt | 83 PF yard : ar e- ks aledonay ding « ohdmag nF do door, door Wieden ie g 1% 6" bpaiy a? = zm Y $ 2 2 Ay Passage Kitchen (wawalty vuuprcd) atmosphere and one often tubercle-laden. These are the reasons for the extensive prevalence of tuberculosis among the native population here and for the relatively high proportion of cases met with among children. A subsidiary but analogous cause is that manner of feeding their infants which many mothers, themselves tuberculous in ‘some instances at least, indulge in, of first chewing the food and then placing it in the mouths of their babies. Further, experiment has shown that there is considerable danger of transmission of tubercle bacilli from one person to another by means of eating utensils, if these are uncleaned. In China the eating together in common, the using of the same general dish, the insertion of individual chopsticks into the food supplied for the meal, the transference of food with the chopsticks of one person to the mouth or dish of another-—all these afford a ready means of conveyance of the organism, but readily preventable when it once comes to be recognised. These, however, are not so important as the one first named—the 222 expectoration habit—for intestinal tuberculosis, as shown in this series, is not the common mode of infection of children out here. Dr. Chaussé compared the infection arising from dried sputum and that from droplets of sputum and saliva directly inhaled, and laid stress on the fact that it is exceptional for those living with tuberculous subjects to receive directly the cough of the latter, and the droplets are very soon diluted with a large quantity of air, and thus danger is lessened. Further, these droplets dry very quickly and their virulence is probably much diminished in ten days. Deposition occurs in a few hours at most, and, owing to the deposition and the drying, the droplets become reduced to the condition of infective dust. ‘Les gouttelettes bacillaires elles-méme, comme facteurs de contagion de la phthisie jouent la plus grande partie de leur réle aprés leur dépét et sous la forme de poussiéres séches, de nouveau mobilisées dans |’atmosphére.’ When no precautions are taken, and this is the rule, the clothes, bedding, etce, are soiled by the virus in large quantity. Then the making up of the bed, brushing of clothes, sweeping the floor, distribute the organisms so that healthy contacts become exposed to infection every day and all day long, for this dust may remain suspended for several hours. Moreover, currents of air, ntovements on the part of the patients, continually put the dust in motion. Then, when the colder months come on the Chinese take out their old clothing which has been stored away in some cupboard or dark corner and wear that over their other garments. When the house cleansing takes place these clothes are not disinfected, so that they are nearly, if not quite, as infectious as when they were laid aside at the termination of the previous cold weather. Nuttall has estimated that a patient with open tuberculosis may expectorate four thousand million bacilli daily. In a dark, shut-in room the chances of infection for a child brought in contact with such a case are enormous. During the crawling age these chances are still greater. It will be seen on perusal of the age table appended that of the two hundred and twenty-five cases under ten years of age, one hundred and sixty-three, or 72’8 per cent., were three years old or less, and one hundred and eighty-five, or 82°2 per cent., under four years. Further, Dieudonné of Warzburg (1903) has proved that the nasal secretion and dirt on the hands of children 223 during the crawling age, say nine months to three and a half years, in a very large proportion showed the presence of tubercle bacilli. The opportunities for the repeated infection of a child of the crawling age in contact with a person suffering from open tuberculosis, or even living in a room previously occupied by such a person, are enormous. The danger of infection from tuberculous members of a family is not, according to the weight of evidence now available, due to the mere ‘tendency to infection,’ or even to the increased probability of infection fev se, but is to be ascribed to the probable large size and frequency of the infecting doses. It has been shown experimentally that small repeated infections may be protective, but large ones overwhelm. Thus is explained the apparent antagonism of Calmette’s and of Bryant’s experiments. The former have already been mentioned with reference to single or repeated infection of cattle, either directly or by keeping them in contact with tuberculous animals. Bryant, after injecting guinea-pigs with a small number (eight) of bacilli daily for a time and then every three days for a period of four months, found lesions indicative of great resistance. The conditions in Hongkong, as has been pointed out, are analogous to the experiments of Calmette. It is needless to labour this point further, but in conclusion mention may be made of ' Theobald Smith’s statement in the Journal of the American Medical Association: ‘The resistance of the tuberculous animal to super- infection is readily broken down by slightly increased dosage, and is successful only when very minute doses come into play.’ As already mentioned, the prevalence of tuberculosis is closely connected with social and economic conditions—overcrowding and slums, poverty, insanitation and squalor. With regard to over- crowding, little, if anything, can be done; the space available is limited, and the population is relatively large and is increasing. The question reduces itself, for practical purposes, to the finding of a remedy for the spitting habit in the first place. The use of food utensils, bowls, chopsticks, etc., in common probably plays a very subordinate part, as has been stated above. To exact, or rather to inflict fines for spitting in public would be useless for several reasons : firstly, the offenders cannot afford to pay fines; secondly, the native police are every bit as bad offenders in this respect as the rest; thirdly, I am convinced that it is the expectoration on the floors of the rooms in which the people live that is the chief source of the bacilli, 224 so that, even if expectorating in public out of doors were stopped— ‘a consummation devoutly to be wished’ on other accounts than the risk of disseminating tubercle bacilli—there would be no deterrent to a man exercising the privilege in his own rooms, unless his wife objected, and this she would not do seeing that she also freely avails herself of a iike privilege. We are, therefore, reduced to the slower but more certain method—Education, and the question naturally follows: How and by whom? European doctors would be of little, if any, use directly. They do not come into contact with the poor to any extent, and they would not be listened to even if they could succeed in making themselves understood. The Chinese doctors would have a better chance, and, therefore, much may be hoped from the locally qualified practitioner who comes more into contact with the people, both at the hospital during his years of training and also after he graduates and goes into practice. Better still would it be to attempt to instruct the people in their houses, and this, I venure to suggest, could be accomplished in the following way :—The Y.M.C.A. and the Y.W.C.A., to which many of the better educated classes belong, have courses of lectures from time to time on various subjects. Like all the laity they are interested in medical questions, and there is no reason why popular (or, to use Huxley’s preferable term, people’s) lectures and — demonstrations should not be given at these institutions, and such lectures cn medical subjects never fail to draw. This would form a nucleus, and from among the members, the women more than the men, some would certainly be found to spread the glad tidings of the gospel of health, at first in their own homes among their servants, and later tactfully to promulgate the doctrines they had been taught. After a time a regular system of district visitors (in the medical sense) might be inaugurated; these would not only instruct the mothers in child welfare and the dangers of feeding their children in the way indicated earlier, but would instil into the adults, and, through them, the growing children health principles generally. The teachers of the Chinese attending British schools might also with advantage incorporate elementary hygienic principles as object lessons for their pupils. Then the question of the establishment of a tuberculosis dispensary would come up for consideration. The Chinese are very fond of their children, and the appalling mortality from tuberculosis must be ascribed to ignorance and not to perversity. 225 +1 1S di ot F1 sivak oz IdA0 [30 IAQ sivad | sivad | “i Lop § ee I oS-oF | of-0f I! sivak O%-O1 (2301, savak oz-S1 val a a gzI 66 6£ Ze 1+ zt ha of sivat O1 0} dn TR30.L It a4 op rai +£ Zr di It Si LE Lt ley gt 6z Lr Il Sz $1 ol “a soxXos 40g “[eI0], _ mn] ** peipuny aasyi FOU S| J | eee paapuny piigy, | SS —* porpung pooseg sivad S-+ *] qavy, sread $-£ sivad -c savak t-f1 sivak Fi-1 *x98 puv ase 0 Jurpsooe saseo OOf ay} fo UONNQUAsIp Surmoys o1 syjuour zI-9 japun 10 syuour 9 ‘Ww X9§ “* paapuny qsity 226 REFERENCES Apvamt, J. G. (1909). Principles of Pathology. Vol. I, p. 184. Burnet, Er. (1915). Annales de l'Institut Pasteur. Vol. XXIX. Cuausse, P. ‘ Nouvelles Recherches sur la Contagion de la Tuberculose par !’air expiré pendant la toux.’ Ann. del Institut Pasteur. Detepine, S. ‘Contribution to the Study of delayed or “latent”? tuberculous infection.’ Annales de I’ Institut Pasteur. FRIEDMANN (1905). Vuirchow’s Archiv. CLXXXI, p. 150. Latuam, A. (1908). Discussion on ‘ Heredity and Disease.’ Royal Society of Medicine. McrapyEaNn, J. (1908). Discussion on ‘ Heredity and Disease.’ Royal Society of Medicine. Nurratt (1911). Quoted by Cautley in Discussion on ‘ Portals of Entry in Phthisis.’ Proc. Roy. Soc. Med. Pearse, W. W. ‘ Sanitary Progress in Hong Kong.’ Scumort, and Bircn-Hirscurerp (1891). Ziegler’s Beitrage. Vol. IX, p. 428. and Kocxet (1894). Ziegler’s Beitrage. Vol. XVI, p. 313- Smiru, THEOBALD (1917). ‘Fournal of Amer. Med. Assoc. Vol. LXVIII. 227 THE PREVALENCE AND CHARACTER OF TUBERCULOSIS IN HONGKONG BY HENRY HAROLD SCOTT (Received for publication 1 July, 1921) II. THE PORTALS OF ENTRY AND MODE OF SPREAD OF TUBERCULOSIS The skin route as a portal of entry for tuberculosis which becomes widespread is negligible; the nose and mouth are the only ones worthy of serious consideration, and from the bacillary point of view a certain amount of exchange between the respiratory and alimentary tracts is possible; inhaled bacilli may be returned by ciliary action of the epithelial cells and swallowed, while the latter | may in turn rise to the level of the larynx and be inhaled. That bacilli when ingested can pass°through the intestinal walls and affect the mesenteric glands without leaving any trace of their passage is a well-recognised fact, and they may also traverse or grow in and be transmitted from these glands to the lungs and elsewhere. Experimental evidence goes to show conclusively that a very much larger dose of bacilli is needed to set up the disease by way of the alimentary tract than by the respiratory. In fact, there is considerable difficulty in infecting animals by feeding. As Cobbett states: ‘The intestine is well guarded. It is otherwise with the lungs. While not open to the attack of common bacteria in anything like such numbers as is the intestine, it (ie., the lung) has not developed equal powers of defence, and tubercle bacilli when they - enter the bronchi can effect an entrance (z.e., penetration) and cause tuberculosis in numbers which would be powerless for harm if swallowed.’ Thus, Gebhardt, working with diluted sputum, found that eight hundred bacilli sufficed to infect by inhalation, while ten to twenty millions failed when swallowed. Kossell, Weber and Heuse found that 1 milligramme would infect calves by inhalation, while one thousand times this dose given by feeding only caused 228 minimal lesions. More recently, Weber and Titze showed that it requires at least 10 mgrms. of bacilli to infect a calf by feeding, while 1/100 mgrm. may succeed when given by inhalation. Findel infected dogs by inhalation with doses down to 014 mgrms., while feeding with doses up to 63 mgrms. produced no effect whatever. Also sixty-two bacilli would constantly infect guinea-pigs by inhalation, even twenty sufficing in some cases, and in very young animals even five. He gave other guinea-pigs doses up to twenty thousand bacilli by feeding, but failed altogether to infect. Many species of animals, including calves, goats, dogs and guinea-pigs, are vastly more susceptible to infection by way of ae air passages than by way of the alimentary canal. The same investigator, Findel, again showed that five million tubercle bacilli when inhaled by a dog caused extensive tuberculosis of the lungs, while one thousand two hundred and twenty times that dose when given by the stomach produced no effect. As regards guinea-pigs, in some experiments as small a number as twenty bacilli were sufficient to cause pulmonary disease when inhaled, whereas the animals were able to withstand three hundred and eighty-two thousand times that dose when introduced into the alimentary canal. Passing from these animal experiments to human cases, an interesting and instructive example was brought forward by Cautley in which one hundred and fifty-one children and two hundred and nine adults were unconsciously subjected to feeding experiments by consuming milk from tuberculous cows. Of these three hundred and sixty individuals ‘ only two of the children were affected, and they merely had a mild adenitis. . . They had taken the milk for a year and a year and a half respectively, and the milk was from cows with virulent disease of the udder. Such cases imply that to secure human infection by tuberculous milk the requirements are: youthful age, a badly infected milk, and the prolonged ingestion of such milk.’ Judging from this and the foregoing experimental work, we may say that the probabilities, when one finds what is apparently a primary lung infection, are that the condition has actually arisen by inhalation and not by way of the digestive tract. A more detailed consideration of the site of origin and the mode of spread in individual cases is given later with illustrations from this series. The tonsils have been regarded by some as the portal of entry 229 for the bacilli, and they quote tuberculosis of these glands in support of their view. “But as there is usually a focus in the lungs as well, it is rarely possible to say whether the primary site of infection was tonsil or lung, or whether both may not be secondary to another portal. As Austin says: ‘Examples of apparently isolated tuber- culosis of the tonsils should be regarded with reserve, as the presence of latent foci in the lungs can never be excluded.’ Excised tonsils of forty-five children (aged 2} to 15 years) examined for tuberculosis by inoculation into guinea-pigs, by histological examination of sections, by cultures and by smears, all proved negative except one inoculation; we can say, therefore, that infection of the tonsils is uncommon alone. Jn fact, in most cases of tuberculosis of the tonsils lesions are found elsewhere, especially in the cervical glands, and it is rare to find tubercle bacilli in tonsils of children without clinical evidence of tuberculosis. ‘By those who hold that pulmonary infection arises through the cervical glands,’ says Dr. Whipham, ‘various routes by which the tubercle bacilli may gain access to the lung have been suggested : first, that from the cervical glands they enter the lymph stream, and by means of the lymphatic vessels enter the venous system, and so are conveyed to the lungs; secondly, that their path is from the cervical to the supraclavicular glands, and thence to the apex of the lung; thirdly, that from the cervical they pass to the bronchial glands, and thence are conveyed to the blood-stream and the pulmonary tissues, though in this connection it must be remarked that ne communicating channels between the cervical and bronchial glands have been shown to exist.’ The usual modes of entry and extension are, of course, the following :— (1) Mechanically with air, food, secretions, and so on, as for example when inhaled bacilli lodge in some spot and produce a focal lesion or are returned by the action of the ciliary epithelium, as has been mentioned already; or a focus having formed, the material bearing organisms finds its way into a bronchus, the bacilli pass out and are swallowed or are drawn into another bronchus to set up the condition afresh there. (2) By direct extension through the tissues with caseation of the parts first affected and involvement of the new tissues by contiguity. (3) By way of the lymphatic channels which become invaded by 230 this contiguous extension. The bacilli thus pass to the next lymph- node, and so on by the lymphatic vessels till the thoracic duct conveys them to the left subclavian vein and the lymphatic mode of spread then becomes (4) Dispersion or distribution by way of the blood-stream ; thence they spread by the right side of the heart to the pulmonary arteries, and so to the lungs generally. Or, by the extension of a focus and erosion of the wall of a blood-vessel, they may pass by way of the pulmonary veins to the left side of the heart and thus to the whole systemic arterial distribution. Or yet again, by opening into a smaller artery will invade the area supplied by that vessel. Cobbett, in his summing up, expresses his opinion in these words: ‘While I have no doubt that tuberculosis is frequently of intestinal origin, especially in children, inhalation is the common mode of infection, not only in phthisis, but in other forms of tuberculosis, especially those in which the bronchial glands seem to be the parts first affected.’ In temperate climates isolated primary tuberculosis of the intestines, rarely seen in adults, is not uncommon in children (estimated at 25 per cent. according to some authorities) ; in tropical countries, judging from my experience in the West Indies (Jamaica) and the Far East (Hongkong), this route, or rather portal of primary entry, is not proportionately so common. Moreover, in temperate climates primary intestinal tuberculosis in children is usually ascribed, if not always traced, to the swallowing of the bacilli in quantities in infected milk. This is certainly not the case here. It is probable that the intestinal infection is in the majority of cases secondary to lung tuberculosis from swallowing the sputum, even in children, and when definitely primary is due to direct ingestion of the bacilli from dried sputum in infected dwellings. With this preliminary clearing of the ground we are in a position now to examine the extent to which these conditions as regards the portals of entry are borne out in my series. Of the whole three hundred cases there were two hundred and nine in which the primary portal of entry appeared to be by the respiratory tract, 7.e., 69°66 per cent., and in seven others which are discussed later there was considerable evidence in support of the 231 same portal, bringing the total to two hundred and sixteen, or 72 per cent. Only thirty-two have been definitely determined as being of alimentary origin, z.e., 10°66 per cent.; five more, discussed below, were very likely alimentary; this would bring the total to 37, or 12°33 per cent. In four other casés there was a possibility of almost simultaneous entrance by way of the respiratory and alimentary tracts. In the remaining cases the primary portal was uncertain. A few remarks are given subsequently on each of these. The number of cases occurring in adults in this series is proportionately small, insufficient at present to form a useful basis for study in detail, and the main objects of this investigation have been the prevalence and character of the disease in children. Of the three hundred cases there were two hundred and twenty-five of ages up to ten years, and each consecutive hundred contained very similar proportions; thus there were seventy-six in the first, seventy-three in the second, and seventy-six in the third. “As regards the sex and ages of those comprising the three hundred, the table (see p. 225) shows that one hundred and fifty-five were males and one hundred and forty-five females. It will be noted that after adult life the cases in males preponderate. Whereas in children up to the age of ten years the proportion of males to females is about as 4 : 5, after the age of twenty years the proportion is as 36: 1. This does not mean that the males are more subject to tuberculosis in adult life than are females. When ill the Chinese exhibit strongly the homing instinct, and make every effort to return to their birthplace to die. The females can more easily do this; the males would if they could, but they in many cases have to remain at work until it is too late. Of the two hundred and twenty-five children under ten years of age there were one hundred and twenty-six females (Z.e., 56 per cent.) and ninety-nine males (44 per cent.). This is not of much significance when we bear in mind that the female child in China is not considered of much account, and among the poorer classes from which these records have been made, the females are less well looked after. This is shown by the relative proportions of sexes for the early months of life. 232 Of the two hundred and twenty-five cases under ten years of age, then, one hundred and forty-eight, or 65°77 per cent., were found in whom the portal of entry was respiratory, and in five others (discussed later) there was a strong probability of a respiratory — origin, in which case the percentage increases to 68. Only thirty-one of these two hundred and twenty-five cases appeared to be definitely of alimentary origin, z.¢c., 13°77 per cent., while among those of ‘ uncertain portal’ were three others in whom the evidence for the alimentary route had considerable weight; if these be included the total percentage would be 15°11. Cases of isolated primary tuberculosis of the intestine are very rare here in my experience. In fact, only four were met with, namely, Nos. 7, 8,81, and 112. The great majority, then, have a respiratory portal of entry, the proportion to alimentary being as great as between four and five to one. In the following the:primary portal was not determined with certainty. It will be of interest to give a brief note on each of them. No. 15. Female, aged 22 months. The extensive spread of miliary tubercles in the lungs, the meninges, liver, spleen, etc., indicate spread by the systemic blood-stream, possibly after the pulmonary circuit, but the only focus found was the caseating right broncho- pulmonary gland; how this became affected without some preceding lung focus I cannot say, but minute search failed to find any. Except for the gland all the tubercles appeared to be of the same age and recent. No. 27. Female, aged 1 year. There were two foci in the lower lobe of the right lung and miliary of the same lung with corresponding glandular affection, the gland at the left hilus probably by extension, the left lung not being affected. The intestine showed tuberculous ulceration and the mesenteric glands were in large caseous masses ; the spleen was the only viscus with tubercles. If the primary portal were alimentary and the spread to the lungs via the thoracic duct, one would expect both lungs to be involved. In favour of the respiratory origin of the lung condition are the foci in the right lower lobe and the glandular involvement. The ages of the pulmonary and the intestinal conditions would appear to be about the same, and there is the possibility of the simultaneous entry by both routes. No. 33. Female, aged 4 years. A few miliary tubercles were scattered throughout the right lung and over the right visceral pleura; the superior tracheo-bronchial gland on the right was enlarged and contained creamy pus; the hilus glands were swollen. The left lung and glands showed no involvement. ‘There were a few miliary tubercles over the spleen surface ; miliary also at the base of the brain, especially about the interpeduncular space. Minute search failed to find any focus, unless the glandular. abscess constituted it 233 No. 34. Male, aged 1 year. In this case the broncho-pulmonary glands and some of the tracheo-bronchial were enlarged and caseous; minute search, however, failed to find any focus in either lung. The mesenteric glands were also, some of them, enlarged and caseating, but not adherent. The state of the mediastinal glands would warrant one in expecting to find a lung focus. In this connection may be quoted the results of experimental work by Calmette, Guérin, and Breton. They found that in guinea-pigs dying in two to four weeks after being fed on the bacilli the mesenteric glands (especially the superior deriving from the small intestine) were enlarged and inflamed, although no trace of any intestinal lesion could be deter- mined. After 6-7 weeks these glands were caseous in greater or less degree and the lungs showed involvement by miliary tubercles with affection of the corresponding tracheo-bronchial glands. Hence in the present case the probability might seem to be primarily alimentary setting up mesenteric adenitis and mediastinal glandular affection secondary to this. N. 35. Female; aged 2 years. Infection extensive: broncho-pneumonic phthisis of the right lung with cavitation in the middle lobe ; the left lung contained miliary tubercles all through, but no focus; mediastinal glands, as would be expected, were more affected on the right. The intestines showed numerous ulcers, one having perforated, and the mesenteric glands were large and caseous; the liver and spleen showed miliary tubercles, the latter more than the former, but the kidneys and brain showed none. The intestinal ulceration being so extensive, together with the condition of the mesenteric glands, favour the infection being primarily alimentary whence extension took place via the thoracic duct to the lungs, though the right lung was more affected than the left. On the other hand there had been time for cavitation to arise so that the intestine might have become involved through the swallowing of infected sputum. Here again the respiratory and intestinal conditions did not appear to differ greatly in age, and it is quite possible that the two-fold route was taken. One must always remember that to infer the relative ages of two tuberculous lesions from the stages present is a proceeding liable to fallacy. No. 39. Male, aged 8 years. This case showed widespread infection though the course of the extension is difficult to trace. The oldest sites would appear to be: (i) The left mastoid which was caseous and necrosed. As there was middle ear disease this may have been a condition apart from the tuberculosis; although caseous and necrotic, there was no actual proof of its being tuberculous in nature. (ii) Chain of large caseous cervical glands, more on the right than on the left. (iii) Thick caseous adherent peritoneum and mesentery as a vast cheesy sheet. (iv) Chain of large caseating mediastinal glands in relation to both lungs. (v) Focus in upper lobe of left lung. (vi) Mesenteric glands in large caseated masses. Other serous membranes—pleura and meninges—showed merely small miliary tubercles, in the latter limited to the base and not very numerous ; the viscera—liver and spleen— showed none on section, merely a few on the surface; the kidneys none, and the intestines none, except miliary on the peritoneal surface. The disease would, therefore, not seem to have been a generalised blood-stream infection. Was the primary portal of entry the intestine, affecting the mesenteric glands and the peritoneum without leaving any trace in the intestine itself, and thence spreading via the thoracic duct to the lungs? Or was the mastoid primary, thence 234 affecting the cervical glands (but the mastoid affected was the left while the glands were more marked on the right), and so by the blood-stream the lungs? Or were the cervical glands affected via the tonsils (they showed no signs themselves), and thence the ear and lungs? Or, lastly, was the primary route respiratory, thence to the mediastinal glands and to the blood-stream, and by the sputum to the alimentary canal ? No. 50. Male, aged 3 years. Focus in the left upper lobe and miliary throughout both lungs ; extensive intestinal affection—ulcers in both large and small intestines, mesenteric glands in caseous masses. Miliary tubercles in liver, spleen, and kidneys, but not in the meninges or brain. The alimentary appears a little older than the respiratory. The question is whether there was infection by double route, respiratory from the focus in the upper lobe of the left lung, and the intestine separately, or whether intestines first, thence by the mesenteric glands to the thoracic duct and lungs and then generalised ; but, if so, why should the meninges not show any involve- ment? Or, again, the lung focus may have been the primary condition, the intestines have become affected from that by swallowing the sputum, and from the intestines the mesenteric glands, the lymphatic system, thoracic duct, superior vena cava, and again the lungs to produce the miliary condition there. N. 52. Female, aged 4 years. The distribution in this case was fairly general but showed certain peculiarities. There were a few minute tuberculous ulcers in an early stage in the small intestine with tubercles on the corresponding serous surfaces. The mesenteric glands, though not large nor adherent, were caseous. The peritoneum and omentum were thick and studded all over with tubercles varying from miliary to the size of large hempseed. As regards the viscera, the liver and still more the spleen showed miliary and grey tubercles, but on the surface only, none were seen on section. A remarkable thing, however, was that the ovaries were both enlarged, almost to the size of adult ovaries, and contained caseous masses, or rather consisted of a caseous mass in each case, the left a little larger than the right. The right lung contained two foci in the lowest lobe, the size of a pea, miliary tubercles throughout, and a more closely-packed mass of grey tubercles subapical in position ; the superior tracheo-bronchial and tracheal glands were enlarged and caseous. The left lung showed miliary tubercles throughout, with closely-packed milia on the somatic pleura; none such were seen on the right. The pericardium also showed several miliary tubercles. There were a few scattered tubercles along and just above the right Sylvian fissure and some at the interpeduncular space. Which was primary is largely a matter of conjecture. For a single organ the ovary was the most extensively involved, each being converted practically into a cheesy, almost creamy, mass. The two lung foci had a fairly hard caseous consistence and were well defined. The alimentary tract tuberculosis and the serous mem- brane involvement, particularly the latter, were extensive, of the miliary type, though, as stated, a few of the glands were caseous. No. 54. Female, aged 7 months. Distribution widespread ; from appearances the respiratory and alimentary involvements seemed to be of about the same age. There was a focus at the lower edge of the right upper lobe, extending a little into the middle lobe; miliary and grey tubercles were present elsewhere in the lungs, and the glands were caseous. Alimentary : ulcers in the small intestine, mesenteric glands adherent and caseous, 235 but not fused ; liver and spleen showed a few miliary tubercles. The right kidney had a small mass occupying the pyramid at the lower pole. The meninges at the . base contained several miliary tubercles. A cervical gland on the right side was enlarged and beginning to break down in the interior. The question is whether the lung focus was the primary (the enlarged gland on the same side would support this) from whence extension occurred to the blood-stream to be generalised and by swallowing of the sputum the intestinal condition ; or, whether from the lung primarily to the intestine and mesenteric glands, and by the lymphatic system to the pulmonary circulation to set up the miliary tubercles there ; or, lastly, Jungs and alimentary tracts together, then generalisation from the lungs to the systemic vessels and from the alimentary tract by the glands to generalisation in the lungs again. No. §5. Male, aged 3 years. The right lung showed tuberculous broncho-pneumonia of the middle and lower lobes ; the upper appeared to be quite unaffected. The middle contained at the lower part a cavity the size of a cobnut. The left lung consisted of three lobes and showed small ulcerated cavities throughout, the average the size of a pea, but rather larger in the middle lobe. Against the lung being primary is the fact that in spite of the extensive tuberculous condition of the organ the glands, though a little swollen and congested, showed no obvious tuberculous infection. On the other hand, the mesenteric glands were enlarged and three of them were caseous, almost chalky, though the intestines showed no change to the naked eye. The possibility is that the alimentary route was primary and thence affecting the lungs by the lymphatic system, the upper lobe escaping, and the hilus and other mediastinal glands not having become tuberculous before death supervened ; or that, since the pulmonary condition was broncho-pneumonic (rather than miliary by way of the pulmonary circulation), the disease had originated by the respiratory _route and the alimentary tract had become involved by swallowing the sputum. No. 58. Male, aged 14 months. In spite of the age of this child the conditions were such that the general distribution of the disease makes it a matter of conjecture as to the primary portal of entry. There were signs of tuberculous broncho-pneumonia throughout both lungs, with ulceration in the left lower lobe. Generalisation by way of the blood- stream may have taken place from this, causing the deposition of tubercles in liver, spleen, kidneys, and brain. The intestinal ulceration and the masses of caseous mesenteric glands may have arisen from swallowed sputum, but the intestinal condition appeared to be equally advanced as the pulmonary and may have arisen independently. The pericardium also was studded with miliary to pin-head tubercles, No. 59. Female, aged 4 years. There was marked kyphosis from extensive involvement of the vertebrae. The spinal site was probably the oldest. From the number of ulcers in the intestines, the large and small both being involved, and the mesenteric glands being in large caseous masses, the alimentary tract would appear to have been attacked prior to the lungs. The latter showed miliary tubercles throughout, but there was also a fair-sized focus (cobnut) just below the apex of the right lower lobe in an early stage of caseation. 236 No: 65. Female, aged 8 months. The peculiar limitation of the disease in the lungs to the middle lobe of the right, which contained a distinct focus the size of a pea and small miliary and grey tubercles throughout that lobe, would be in favour of a primary (or at least independent) lung involvement, the alimentary tract being affected secondarily from swallowing the sputum. The mesenteric glands, however, were in large caseating masses and appeared to be of older standing than the pulmonary condition. No. 66. Female, aged 20 months. In some respects similar to the last. In this case the lung affection was limited to the right upper lobe in which there was a subapical focus, and in addition generalisation of small tubercles throughout that lobe. The disease was much more advanced in the alimentary tract—intestinal ulceration and large caseous masses of mesenteric glands—and this would point to an alimentary portal of entry. It is not improbable that the lung infection, though later, was independent of the intestinal. The condition of the brain (cerebellar mass) and meninges (miliary to hempseed on vertex and base) point to dissemination by the blood- stream, but no other viscera showed involvement, to the naked eye at least. No. 68. Male, aged 3 years. This case is remarkable in that, except for a hilus gland and a paratracheal gland on the right side as large as a cobnut and filbert respectively and caseous, no focus was found after careful search. There were sparse miliary tubercles throughout both lungs; in the meninges they were numerous, especially at the base, the interpeduncular space, the ependyma, along the Sylvian fissures, and a few on the vertex. ‘The spleen showed also a few milia on the surface. There was no evidence of tuberculous affection of the intestines. No. 70. Female, aged 7 years. In this child there was a mass of caseating glands on both sides, involving the inferior and superior tracheo-bronchial and the paratracheal, more on the right than the left ; the broncho-pulmonary were not apparently affected. No focus was found in the lungs. The tonsils were not affected, nor the cervical glands, and the spine was examined without success. The alimentary tract was free. In the kidneys, however, there were in the left miliary and grey tubercles, not numerous ; in the right more, and arranged almost focally as a group at the base of a pyramid towards the lower pole. No. 92. Male, aged to years. The primary portal in this case was probably alimentary. There were several ulcers, more in the large than small intestine, however, and a thickened tuberculous mass like a collar at the ileo-caecal valve. Mesenteric glands were enlarged, in masses, and caseating. The lungs showed tubercles varying in size from pin- point to small pea throughout, but the mediastinal glands did not show any tubercles visible to the naked eye, and only one was a little congested and swollen. Neither brain, liver, nor spleen showed any signs, but the kidneys were severely diseased. The right had the pyramidal areas hollowed and caseous; they had run together and were lined by thick tuberculous matter. The left was in a similar condition but to a less degree; calyces opening into each other with a lining of tuberculous pus. The tuberculosis was, therefore, widespread, but if by way of the blood-stream it is peculiar that the kidneys should have been 237 extensively affected while the brain, liver, and spleen escaped. The lung condition appeared to be more recent than the intestinal and may have arisen by way of the pulmonary circulation as previously pointed out. No. 95. Femaie, aged 8 years. : In this case one of the inferior tracheo-bronchial glands was caseous and one of the superior in an earlier stage, both on the right side, but minute search failed to find any tubercles in the lungs. The intestines were not affected, and no focus was detected anywhere. The meninges showed miliary tubercles in fair numbers at the base, fewer along the Sylvian fissures, and only an occasional one at the convexity. No tonsillar or retro-pharyngeal affection was detected. If the meningeal condition arose by inhalation, via the ethmoidal canals (as meningococcus may pass), then we have no explanation of the mediastinal gland affection. Anyway _these do not appear to be accounted for, though the spread may have been from them by the blood to the meninges. No. 96. Female, aged 9 years. In this case both the respiratory and alimentary tracts showed such extensive involvement that the only doubt is whether the lungs, which were in a state of tuberculous broncho-pneumonia throughout, with cavitation in the right upper lobe and considerable ulceration in the middle lobe, were the primary seat and the intestine was involved later through swallowing the sputum, or whether the alimentary tract formed the primary portal of entry. The small intestine contained tuberculous ulcers; the mesenteric glands were in large caseated masses and adherent, and the peritoneum was studded with tubercles. Of course, there may have been infection by both routes nearly simultaneously. No. 100. Male, aged 5 years. In this child there was a cavity in the right upper lobe the size of a medlar, with grey tubercles surrounding it and smaller ones at a greater distance. No other part of the right lung was affected and the left showed nothing. There were pleural adhesions at the apex and direct communication with a tuberculous ulcer the size of a crown-piece above the right clavicle. Whether a supraclavicular gland had become infected from the lung direct and had then opened on the surface, or whether there was a primary affection of the skin which ulcerated and extended to the lung is doubtful. “There was a tuberculous skin affection with early ulceration on the right cheek also. No other viscera showed any involvement, and no cervical glands were found enlarged. This may be an example of the mode of spread already mentioned. No. 104. Male, aged 16 months. In this the disease was widespread and the primary portal and mode of extension doubtful. The left lung showed two foci, one subapical in the lower lobe, the other nearer the lower edge, and each. was surrounded by a zone of small caseating tubercles. The right upper and lower lobes contained miliary tubercles; the ‘glands, hilus, and tracheo-bronchial were caseous and enlarged, In the alimentary tract there was nothing, but one or two mesenteric glands contained small foci. Milia were present in the liver, more and larger in the spleen, The meninges had masses of miliary tubercles at the base, and along the Sylvian fissures and in the interpeduncular space; a few on the vertex and internal surfaces of the cerebral hemispheres. The spine showed a large abscess the size of a Tangerine orange, involving the 11th and 12th dorsal and the 1st lumbar vertebrae. Sug- 238 gested method of spread: Lung (left), then by inhalation the right, by the blood- stream to the liver, spleen, spine (injury possibly determining this site), and also by the sputum to the intestine and mesenteric glands; or, from the left lung foci by the sputum to the intestines and mesenteric glands, thence by the lymphatic system to the heart and pulmonary circulation, causing the miliary tuberculosis -of the right lung, and also by way of the blood-stream from the lung to the liver, spleen, and meninges. But the spine focus must have been of considerable standing, for there was marked caries and an abscess formed containing caseous matter. No. 116. Female, aged 10 months. The right lung showed two distinct foci in the upper lobe : the larger subapical the size of a cherry and caseous, the smaller the size of a pea, near the interlobar fissure ; the rest of the lung contained miliary tubercles. The left lung showed miliary to pin-head tubercles, sparse in the upper lobe, more in the lower. The mediastinal glands were enlarged, with caseating foci. The mesenteric glands (the upper set) were in a similar condition. The question is whether both the respiratory and alimentary tracts were involved simultaneously, or whether the lung focus was primary, the alimentary secondary, passing to the glands without leaving signs in the intestine itself. Thence by the lymphatic to the pulmonary circulation to set up the miliary tuberculosis there, and from the lungs via the blood-stream to the meninges, liver, and spleen. But, if so generalised, one would hardly expect the kidneys to escape altogether. No. 118. Female, aged 3 years. In this case the disease was exceedingly widespread and the portal of entry is a matter of conjecture. There were tuberculous ulcers of the right cheek, jaw, and neck; the right parotid, the submaxillary and cervical glands on both sides, were caseous. There was tuberculous broncho-pneumonia throughout both lungs, the pleura showed many tubercles at the upper part of the somatic layer and all over the visceral ; the pericardium was also affected. The peritoneum was studded with tubercles, miliary to pea, and the intestines contained a few tuberculous ulcers, both in the large and the small; the mesenteric glands, both upper and lower series, were enlarged, adherent, and caseated. The liver showed a few miliary, the spleen more and somewhat larger. In the right kidney was a small mass of aggregated tubercles at the lower pole, while in a corresponding situation in the left was a definite caseous focus the size of a cherry. There were aggregated miliary tubercles on the basal meninges, a few also on the vertical. No. 137. Male, aged 10 months. Disease was widespread and severe in this child, especially when one considers the age. The lungs showed a cavity in the lower lobe, a caseous focus in the middle, and miliary tubercles throughout the remainder of the right; in the left there were two caseous foci the size of a pea in the upper lobe, a caseous mass as large as a walnut in the lower, and grey tubercles through the rest of the lung. All groups of the mediastinal glands were involved. The mesenteric were enlarged, caseous, and matted, but appeared to be more recent than the thoracic. There were two tuberculous ulcers in an early stage in the small intestine. The femoral glands on each side were the size of a small walnut and caseous (the origin of these could not be discovered). ‘The cervical glands on each side were caseous, especially on the right. The right kidney contained a small aggregation of tubercles at the lower pole ; the liver a few miliary tubercles, the spleen more and larger. The meninges were not affected. As the respiratory and alimentary tracts were both 239 involved, and also some glands unconnected with either, it is difficult to say which was the primary portal, but the lungs would seem to be most involved and of oldest standing. No. 141. Male, aged 10 years. . The oldest site in this case was to the right of the sixth cervical vertebra. The bone was carious, and there was a sac with pus discharging from it along the spine into the right pleural cavity (it may possibly have started as a retropharyngeal abscess). The right lung showed miliary tubercles throughout, not very numerous, but more in the upper lobe than in the other two. The left lung had none. There was a caseous gland at the right hilus, Tuberculous sinuses were present in both femora and in the left tibia. No. 145. Female, aged 2} years. Judging from the appearance, degree of caseation, etc., the primary portal -may have been alimentary, but the disease Was very widespread, the brain showing several foci. ‘There were tuberculous ulcers in both-large and small intestines, and the mesenteric glands were in adherent, caseous masses. The cervical gland infection was of fairly long standing also, those on the right containing creamy pus, those on the left being in a caseated stage. Both lungs. showed grey tubercles, and the mediastinal glands were in caseous masses at the hilus and along both ‘sides of the trachea. The brain showed: three distinct foci in the left lobe of the cerebellum, another in the right hippocampus, another in the left superior frontal, all these about the size of a pea or a little larger, and in the right supra- marginal gyrus one as large as a cherry—an extensive series of brain foci. No. 149. Female, aged 3 years. It is possible that the alimentary portal was the primary as this tract showed the greatest involvement (not a very safe indication, it is true). There were numerous ulcers in both large and small intestines, the mesenteric glands, both upper and lower series, were enlarged, adherent, and caseous. The lungs contained closely aggregated miliary and grey tubercles throughout, but no focus, although the mediastinal glands on both sides were caseous and the paratracheal on the right contained creamy pus. The brain was considerably involved: there were a few miliary tubercles in the basal meninges, many along each Sylvian fissure, and a few on the opposed surfaces of the cerebral hemispheres ; in the cerebellum at the fore part of the left hemisphere on the upper surface there was a ‘ solitary ’ tubercle, as large as a cherry, and towards the posterior another focus the size of a haricot: on the under surface of the right hemisphere was one the size of a filbert situated anteriorly, and at the posterior extremity another the size of a pea. As regards the course, it does not appear likely that the large tubercles in the cerebellum would be of the same standing as the miliary in the meninges. The intestinal appeared to be the oldest, the lungs probably secondary to this via the pulmonary circulation, for the distribution was equal over both lungs which were granular everywhere from miliary and grey tubercles. General blood infection was supported by the presence of a few tubercles in the liver, spleen, kidney, and meninges, and probably an earlier blood infection set up the various brain tubercles. No. 157. Male, aged 5 years. Lungs were granular in appearance from the presence of numerous grey tubercles. A gland at the right hilus was swollen and congested, but showed no signs of tuberculosis init. Intestines nil. One mesenteric gland, the size of a pea, 240 contained small grey points. Both kidneys showed miliary tubercles, the left a little more than the right. There were milia also in the liver, spleen, and meninges. No focus could be found. The universality and evenness of the pulmonary distribution point to a haemic infection of the lungs rather than a respiratory. No. 159. Female, aged 8 years. The condition in the lungs and in the abdomen appeared to be of about equal development. There were two foci in the left lung, one in the upper lobe the size of a small cherry, caseous with a fibrous border, another in the lower lobe slightly larger. The right upper lobe showed tuberculous broncho-pneumonia, the middle and lower many grey tubercles. The mediastinal glands on both sides were enlarged and caseous. The alimentary tract showed several tuberculous ulcers in the small intestine, and the corresponding mesenteric glands were in caseous adherent masses. There is apparently equal evidence for respiratory and alimentary portals, and the stage in each is about the same. Possibly the double route was taken nearly simultaneously. No. 160. Male, aged 3 years. Miliary tuberculosis of the lungs and meninges, a few also in the spleen. Two paratracheal glands were swollen and contained two small caseous foci, and one at the hilus was enlarged and caseous. No focus was found anywhere. No. 164. Female, aged 6 years. The difficulty in this case is to decide, in view of the widespread condition, which was the primary portal of entry. There were old-standing ulcers beneath each ear, with caseated and breaking-down cervical glands. The right lung showed several foci, certainly not of recent production, one the size of a pea, deep in the upper lobe, another similar at the base of the lowest ; a small cavity in the middle, and, at the upper part of the lowest, a large focus the size of a cherry, caseous, with surrounding tubercles and early ulceration. In addition to these there were milia throughout the lungs. There were tuberculous ulcers in the intestines ; the mesenteric glands were enlarged, caseous, adherent. The meninges showed numerous tubercles at the base and along the Sylvian fissures, more again below the cerebellum. The vertex and inter-hemispherical aspects showed tubercles in fair numbers. In spite of the lung condition the mediastinal glands, though containing tuberculous foci, were not nearly as much affected as the mesenteric. The respiratory and alimentary appeared to be of nearly equal age, as regards the foci; the miliary state of the lungs may be a secondary result of the intestinal. Doubtful whether: (i) Focal pulmonary, then intestinal, mesenteric, generalisation in lungs and thence the brain; or (ii) Primarily skin, then glands, blood-stream and generalisation; or (iii) Skin, lungs, intestines (the last two independently and simultaneously, as regards the focal state), and subsequent generalisation. No. 178. Female, aged ro years. Miliary tuberculosis of both lungs, fairly numerous and evenly distributed, but rather more in the right lung. A gland at the right hilus and one paratracheal caseous. Nothing in the alimentary tract. A few miliary in the meninges, more at the base, but some also at the vertex. No focus detected anywhere. 241 No. 181. Female, aged 4 years. Meninges showed general involvement, more aggregated at the base, in the interpeduncular space, along the Sylvian fissures, and below the cerebellum ; few on the convexity. The lungs showed widespread miliary tubercles, more numerous in the right than the left. Mediastinal glands on the right were enlarged and caseous, but no focus was found. No, 203. Male, aged 5 years. Miliary tubercles scattered in moderate profusion and evenly through both lungs, but the source of these was not detected. The alimentary tract did not show any involvement, but one mesenteric gland was the size of a pea, and on section contained a minute caseous ‘point. There was extensive tuberculous meningitis ; at the base they were closely aggregated and pus was forming; there were also many along the Sylvian fissures and only a little less on the convexity. The ventricles contained more than the normal amount of fluid. No focus was found. No. 206. Male, aged 16 months. Grey tubercles in the upper lobe of each lung and less in the right middle lobe. The spleen showed a considerable number of miliary tubercles. The meninges were extensively affected ; there was pus at the base with very numerous tubercles, also along the Sylvian fissures, in the velum interpositum and below the ~ cerebellum. No focus was found, the other viscera yielding no evidence. No. 210. Male, aged 5 years. Here again no focus was found. The right lung contained miliary tubercles in the middle and lower lobes, and a gland at the hilus was caseated. The upper lobe of the left lung also contained miliary tubercles ; there was no affection of the glands on the left; a few miliary were seen also in the spleen, while the meninges were extensively involved : tubercles were numerous at the base and along the Sylvian fissures, and there were one or two on the convexity and on the opposed surfaces of the cerebral hemispheres. No. 211. Female, aged 5 years. Apparently a haematogenous infection, but the primary focus was not found. The lesions were miliary tuberculosis of both lungs, the spleen, and the basal meninges. ‘No. 215. Female, aged 13 months. The disease in this child was extensive and the decision as to the primary portal of entry a matter of opinion. There was a caseous focus as large as a filbert in the right lung, and grey tubercles throughout both lungs, while all groups of the mediastinal glands were enlarged and caseous. The cervical glands were a little affected. The mesenteric were in adherent caseous groups of about the same degree of involvement as the mediastinal ; there were miliary tubercles of the liver and spleen, and a terminal meningitis. The mesentery and peritoneum generally were studded with tubercles. From the state of the glands in the thorax and abdomen there is no evidence for priority. The lung condition with a caseous focus of such a size must be of considerable standing, whereas the intestine itself showed one small ulcer only. On the other hand, the extensive affection of the peritoneum indicated that the abdominal condition was not very recent. On the whole, unless we regard the respiratory and alimentary as about coequal 242 in age, the most satisfactory explanation would be, the lung focus primarily, alimentary secondarily, thence the peritoneum and mesenteric glands, and via the thoracic duct to the pulmonary circulation and so setting up generalisation there, and via the blood-stream (systemic) to liver, spleen, and meninges. No, 222. Female, aged 18 months. Though the mediastinal glands on the right side were enlarged and caseous, no true focus was present in the lungs. The middle and lower lobes showed grey tubercles of the minute bronchioles, while the right upper and both lobes of the left contained miliary tubercles by the blood-stream. Some of the mesen- teric glands were the size of a small pea and had caseous points. The liver, spleen, and kidney contained tubercles varying from small miliary to hempseed, and the base of the meninges showed them also. On the whole the sequence was probably : Respiratory first, causing the broncho-pneumonia of the lower lobes of the right lung, then alimentary to the mesenteric glands, without leaving signs in the intestine itself ; thence by the lymphatics to the pulmonary circulation and so miliary generalisation in the lungs, and from there to the liver, spleen, kidney, and meninges. No. 223. Female, aged 4 years. Extensive tuberculous meningitis, but no focus detected anywhere. No. 227. Female, aged 3 years. Lungs extensively affected with miliary and small grey tubercles, the right rather more than the left. There were numerous tubercles in the meninges, and a few in the liver and spleen, but no definite focus or indication of the primary portal of entry. No. 230. Male, aged 3 years. Tuberculous meningitis most severe, but the origin of this could not be traced. No focus was found, but there was miliary tuberculosis of the lungs, liver and spleen. No. 239. Female, aged 3 years. In this case the alimentary, as evidenced by the state of the mesenteric glands, would appear to be the oldest ; on the other hand the caseous focus in the left lung would be of older standing than the miliary and grey tubercles scattered generally throughout the lung. Also the mediastinal gland corresponding to the lung focus was completely caseous, like the mesenteric glands. The cerebellar’ focus was not very recent, less so than the miliary in the lungs and meninges. There are three possibilities: (i) Primary lung focus, alimentary secondarily, then generalisation in the lungs and from there to the meninges (if so, whence would the ‘ solitary’ tubercle in the cerebellum arise, unless by separate earlier metastasis from the lung focus ?). (ii) Primary intestinal, secondary focus in lung and spread from there by inhalation to other parts of the lung, and by blood- stream to the cerebellum and meninges (but the cerebellar is a caseous node, while the meningeal affection is miliary and apparently recent). (iii) Lung and alimentary routes about the same time (judging from the glandular infection those in the thorax and those in the abdomen appear to be about the same stage), then from the lung to the cerebellum (focus), and from the mesenteric glands to the right side of the heart for the production of the miliary and grey tubercles in the lungs. 243 No. 275. Female, aged 6 years. The lesions found were a general miliary infection of the right lung, and a few milia in the upper lobe of the left; an enlarged and caseated paratracheal gland on the right; sparse miliary tubercles in the spleen, and fairly thickly- studded miliary infection of the meninges, more aggregated at the base. No focus was discoverable, unless the mediastinal gland be regarded as such, but no evidence of the-communication of this with a pulmonary vessel could be made out to account for the distribution in the lung, and there was certainly no focus in the lung to which this gland could be attributed. No. 283. Male, aged 4 years. The lungs in this case presented miliary tubercles, rather more in the right than the left, but not very numerous in either; the hilus on the right showed a large caseous gland, and one paratracheal on each side was caseated, that on the right being the larger. No focus was found in the lungs to account for these glands. There were a few miliary tubercles in the liver, more in the spleen. The meninges were severely affected, at the base and over the right hemisphere. In the latter, just below the cortex of the angular gyrus, was a mass of tubercles focally arranged, aggregating to the size of a large pea or small marble. The cerebral condition had probably arisen by vascular spread from the mediastinal gland, but the source of the latter was not found. No. 291. Female, aged 4 years. The only focus detected in this case was a caseous mass occupying nearly an entire pyramid at the lower part of the left kidney. There was miliary tuber- culosis of the lungs, but no focus. No infection of the intestines; only one mesenteric gland showed anything and this was not so large as a haricot, but had a small caseous point. There were also a few milia in the liver. The kidney lesion was to all appearances the oldest, but whence this arose could not be discovered. REFERENCES Austin, R. S. (1919). American Fourn. of Children’s Diseases. Vol. XVIII, pp. 14-19. Carmette, Guérin, and Breton (1907). Ann. de I’Inst. Pasteur. Vol. XXI, No. 6. Cautey (1gt1). Discussion on ‘ Portals of Entry in Phthisis.’ Proc. Roy. Soc. Med. Cozsetr, L. (1911). Discussion on ‘ Portals of Entry in Phthisis.’ Proc. Roy. Soc. Med. Finver. Zeitschr. f. Hyg. Vol. LVII, p. 83. Kossett, Weser, and Heuss (1905). Tuberkulose-Arbeiten aus dem kaiserlichen Gesundheit- samte, Berlin. 5th Heft. Weser and Tirze (1910). Tuberkulose-Arbeiten aus dem kaiserlichen Gesundheitsamte, Berlin. toth Heft. Wuirnam, T.R. (1g11). Discussion on the ‘ Portals of Entry in Phthisis.’ Proc. Roy. Soc. Med. 7 £48 - ors *) | ’ HHA! tga oud ke s Oar fes0235 wae mat fray 28 SA 3M ody dy otal or Gan sms), att, sid 0! 7yclliens sina oe Tv a4 = see s } ly } # - aah ey 2 TM APRA 4? Io fonoptal W tiem r is ine 7 " ‘ p 4 Pedftaelisat oA} pest wilh types t ia an fe! Auiiwainn ieite® ’ jwit d Gf abr toats mt 4 * pe bi t - Fide } rer TA oa Re hanes Cay eareiae fe ' siat , F A 93, gts ge ewece dh Io wy sdf weekend oF 3h Osgoraee t coe yindoty her? apie ~ 4 ens : bowt hiakawe ' Jieted) i aig aad aoe 4 beye phtmaae waa ifs. of hbiesta cst aithidatel adactor suey iment B 1 bie a Rp? inh ove gycel aaa “yest a » é aa , n ee | ryerd aire hy f im: wal 4 04 neve oral see - ai Eis ads ioe’ as b.4 u . 7B Tas, > SAL ai Verdes | lo elaine be =: aaa Ay et mal Seer Setat) weet fall On Se a ‘winery nid ’ ty GR, Med PE, woh SES a ar > pa i > i ae i ee is uedtist ‘ eictty¥ow) os ; e (fh Ated eteY “ 3.) » “ y F ‘mag es 094 Me taee 245 MULTIPLE ANEURYSMS IN A CHILD BY R. H. KENNAN, M.D. (Received for publication 3 January, 1921) The patient is a well-grown, intelligent, active but anaemic boy of four years old; the son of observant, healthy white parents, American Missionaries on leave from Sierra Leone. The boy had had about six attacks of fever, each of short duration, in Freetown, and the latest of these occurred in October last. His tongue was clean and moist and without tremor; his gums healthy; his pulse regular and of good quality, with frequency of ninety-two beats per minute while he was in the erect posture. His heart was free from murmur and normal, and no enlargement of his liver or spleen or lymph glands could be detected. ‘There was no sign of rickets and his urine (acid, 1,022) was free from albumen and sugar. His parents suggested that there was oedema of his eyelids, but at the time of examination (about 4 p.m.) I could not satisfy myself that oedema was present. The upper end of the left hypothenar eminence was slightly enlarged, and prominent, and was the site of a pulsating tumour which could be ‘emptied’ by pressure over it, and which then filled again ‘per saltum.’ Pressure over the ulnar artery greatly diminished, but did not entirely obliterate pulsation in the tumour. The surface tissues over the tumour were unaltered in colour and texture. Just above this tumour, and separated from it by a narrow surface depression, was a second one of similar character. There was no history of injury antecedent to the first hypothenar tumour, which was first noticed in March, 1920; the second swelling appeared in October, 1920: the parents state that it came quickly two days after a fall, which, however, did not cause bruising or damage to the skin. Subsequently a small pulsating little vessel, slightly bluish in colour was noticed about the middle of the right 246 side of his neck, and some time after this, while his mother was examining his neck, she found another larger pulsating, uniformly oval swelling a little lower down, occupying the greater portion of the base of the anterior triangle of his neck above the clavicle. ~The skin over it was freely moveable, as was also the tumour from side to side, but it was not capable of displacement vertically. Vigorous pulsations were easily observabl® by sight and touch in the little vessel; the pulsations could be stopped by slight pressure below the point where it lay immediately beneath the skin. The larger, lower tumour could not be emptied by pressure over it, but the pulsations, synchronizing with the ventricular systole, appeared to be distensile in character, and with a stethoscope a loud bruit was audible over it. The radial pulses were of equal volume, and no difference in colour or temperature of the hands could be detected. It could not be decided whether the wrist and hypothenar tumours were separated by normal vessel or whether the surface depression was due merely to compression by fascial bands. None of the tumours caused any pain, and only when the larger neck tumour was firmly pressed in an effort to empty it did the patient show any sign of distress. Examination of the blood showed: Plasmodium falciparum infec- tion and also eosinophilia (about 12 per cent.). Several very careful and prolonged examinations of the faeces yielded negative results. There appears to be no doubt that the two distal tumours are arterial aneurysms, and that both the abnormal conditions in his neck are of the same kind. The alternatives of ‘venous pulsation,’ and ‘enlarged lymph gland with transmitted pulsation’ were borne in mind at the time of the examination. Macfie and Ingram (1920) have described ‘ Three cases of Cardiac Aneurysms in Native Boys of the Gold Coast,’ and discuss the question of aetiology, and the suggestion is made that malaria may have been a factor in causation of the aneurysms. Aneurysms of the large arteries have been frequently reported as having occurred in young children, even as early as in foetal life, but in the present case not only was the child four years old, but the aneurysms were several and their etiology obscure. Stress has been laid by many on the effect of septic emboli in weakening the artery wall by inflammation spreading from the infective clot in causing 247 aneurysms in children suffering from septic endocarditis, but no such cause was operative in this case. Syphilis is also commonly considered a cause, but though no Wasserman blood test was made on this boy, it is extremely improbable that syphilis was present. Though there was a history of a fall antecedent to the appearance of one of the tumours in this case the relationship of the two things as cause and effect was not definite, while in the cases of the neck tumours any competent injury must have been obvious at the time it occurred. As causes, it appears that septic emboli, syphilis and injury were not present in this case. At one time, the presence of four arterial dilatations in a child of four years old would have been held to justify the non-illuminating diagnosis of ‘aneurysmal diathesis.’ REFERENCE Macrig, and INGRAM (1920). Three cases of Cardiac Aneurysm in Native Boys of the Gold Coast. Ann. Trop. Med. and Parasitol., Vol. XIV, p. 147- ts a : ! cart | eesal Pa Ua “ay maobe Te cp mol Prag yo Nile ere. x? ailiday adt of % Sa i Var ff ny ened 4nd ota Bi , } i de ul) ion mHK9, oF it 08 i“ ' ¥3I0 i 5 A fy ( ” A »,, 0%, 9436 wit > Pitipest CTA 3 od pieiloh) som are Lal bas eri 42s yale 1 1OAfS ORE 9 ae yes Le, § . r. : Seuid ei/ht ik tee Mh, Aap 153 Dap Mer’ 7 0 vastae al | oly Ri rd Let naod wud Fulani, : ite wii re beds (Ay ing +> yr is. ATG riety : 1 «ld Q BRE ecb a by: 7 a om mt nee Tae Wi ey en weal bf Ju¥ ‘ween Gee Doll nt ooh : oP aah ye hei ry J - ; ‘ : ‘ bie 2 wrt ¢ . y he - ' , ' & ee ks att werk We Sy eee 249 LAPPETED ANOPLOCEPHALA IN HORSES BY WARRINGTON YORKE AND T. SOUTHWELL (Received for publication 22 July, 1921) PLATE XVII. I. ANOPLOCEPHALA RHODESIENSIS, nom. nov. About two hundred and fifty specimens of this parasite were collected by one of us in 1912, from eight zebra (Eguus burchelli) in North Eastern Rhodesia. TECHNIQUE. Most anatomical details were easily elucidated by the following procedure :— The werms were stained, ez masse, for several days in dilute acetic-acid-alum carmine, then washed and taken through the alcohols into clove oil. As soon as the worm was clear, it was placed under a binocular microscope and segments were detached, one at a time, by means of a surgical needle or cataract knife, beginning with the posterior segment. The segments were then mounted serially with the anterior surface upwards. This procedure was quite simple until about segments 10 to 15 were reached, when it was found that the segments were so small and close together that the detachment of single ones was difficult and tedious, and also often unsatisfactory. Anterior to this point sections were therefore cut with a microtome, when required. Horizontal sections were necessary, however, to determine the structure of the uterus, e.g., the presence, or absence, of anterior and posterior outpocketings, and certain other details. EXTERNAL ANATOMY. The largest specimen measured 114 mm. long, 22 mm. broad and 3°25 mm. thick, and the smallest 14 mm. long, 4°25 mm. broad and 250 1°75 mm. thick. Specimens less than 30 mm. long were immature. Most of the larger worms approximated the following dimensions :— Length 90 mm., breadth 20 mm., thickness 3 mm.; number of segments, about two hundred and fifty. The following measurements show the size of our largest worm as compared with measurements made by Fiihrmann and Collin of the same species :— Long Broad Thick Fihrmann ... see .. 50mm. I8mm. 3mm. Collin as xs .. 7Oomm. 26mm. 5-6mm.? Our specimen Bee .. 114mm. 22mm. 43.25 mm. The smallest specimen contained ninety segments, and the posterior extremity was much narrower than the middle of the worm, and was rounded; the last few segments were longer (0'25 mm.) than the posterior segments in full-sized worms. A considerable number of Oestrid larvae (Gastrophilus sp.) were found firmly attached to many of the worms. Head. The head is always conspicuous; it is cuboid, with a truncate anterior extremity; the breadth is considerably greater than the length. In large worms measuring go mm. the head is about 2 mm. long and 3°25 mm. broad. Suckers. The suckers, which are situated on the anterior surface of the head, are directed straight forward. They are about 0’7 mm. in diameter, and their cups are surrounded by a definite muscular rim having a diameter of about 200”. The suckers were separated from one ancther by more or less well marked grooves. Lappets. Immediately behind each sucker is a large lappet. The length and breadth of the lappets varies considerably, possibly owing to contraction during fixation, but when they are fully extended they measure about 1°25 mm. long, 1°75 mm. broad, and the distance between the bases of the two dorsal or two ventral lappets is about 0°6 mm. In some specimens the lappets were globular and filled with liquid. Segments. The segments are very shallow and are imbricated. Posteriorly, the worm increases gradually in size, the greatest breadth being usually about 1 cm. from the posterior extremity; it must be remarked, however, that the general shape of the worm varies considerably in different individuals, possibly owing to different degrees of contraction during fixation (Plate XVII). 251 INTERNAL ANATOMY. Muscular system. Both the longitudinal and transverse muscles are strongly developed, and in transverse sections of mature segments the thickness of the former is about 130n, and that of the latter about 60m (fig. 3). The dorso-ventral muscle bands are not so well developed. A short but powerful muscle connects the internal extremity of the cirrus pouch with the transverse muscles of the ventral surface. Nervous system. There are three longitudinal nerves on each side, the median being far the most prominent. The other two, dorsal and ventral, are slightly lateral to the median nerve and close to the transverse muscular layer (figs. 1 and 3). Excretory system. The water vascular system is enormously developed. On each side there are two vessels, a very large ventral vessel having a diameter of about 35m, and a much smaller dorsal vessel which appears to be interrupted from time to time. The ventral vessel is internal to the dorsal vessel which lies over the median nerve trunk. The remarkable development of the water vascular system is one of the most striking features in sections of the _worm (figs. 2 and 3). Genitalia. At least the whole of the posterior half of the worm is sterile, exhibiting no trace of genitalia, and, as in none of the specimens examined was there any évidence that segments had been shed, it is probable that the worm is passed entire in the faeces. Testes. These first appear about segment 4 or 5, and they disappear about segment 25. They attain their maximum develop- ment between segments 11 and 15, where each testis measures from about 55 to gou by 30n to 70n. They occupy the entire medullary parenchyma between the ventral excretory vessel on one side, and the cirrus pouch on the other; they never cross the ventral excretory vessel. They usually lie three or four deep in the dorso-ventral direction, but the larger ones may be only one or two deep (fig. 1). Vas deferens. After running laterally from the testes for a short distance, the vas deferens dilates into the outer seminal vesicle, which usually lies ventral to the cirrus pouch, although it was observed occasionally to lie dorsal or median to the latter structure. It then narrows and enters the cirrus pouch, where it again dilates to form the inner seminal vesicle. The cirrus pouch is remarkably 252 long, and passes dorsal to the ventral water vessel and median nerve to reach the-edge of the segment (figs. 1 and 2). The cirrus is long, slightly coiled, and is armed with very minute spines. Packets of spermatozoa of various shapes and sizes, but usually oval with pointed extremities, are abundant in both the outer and inner seminal vesicles. In the majority of mature segments, the measurements of these various structures are approximately as follows : — Length of outer vesicula seminalis 66 56 eee 550u Greatest breadth of vesicula seminalis ... ie ee 150" Length of inner vesicula seminalis wid Sp ae 700" Greatest breadth of vesicula seminalis ... oa se 20044 Total length of cirrus pouch... Ms a ... 1,000/ Greatest breadth of cirrus pouch a re B% 100m DORSAL VENTRAL Fie. 1. A. rhodesiensis. | Segment, viewed anteriorly, showing male genitalia. _c.s., cirrus sac; e..S., external vesicula seminalis ; i.v.s., internal vesicular seminalis ; /.m., longitudinal muscles ; i.., lateral nerves ; m.n., median nerve; 7.S., receptaculum seminis; 1#., testes; v., vagina; ¥.é.0., ventral excretory vessel; v.g., vitelline glands. 40. Ovary. This first appears about segment 24 and disappears about segment 34; it attains its maximum development about segment 30. The poral wing has a lateral diameter of about 1 mm. and the aporal wing of 2°2 mm. The median axis of the ovary is slightly cn the pore side. The ovary consists of a series of vertical, club-shaped columns arising from a ventral horizontal base. The largest column measures about 170 dorso-ventrally and 60,2 laterally. The columns decrease in size towards the periphery of the ovary (fig. 2). In the antero-posterior direction they are never more than two deep. 253 DORSAL tm. VENTRAL Fic. 2. A. rhodesiensis. Segment, viewed anteriorly, showing female genitalia. c., cirrus ; ¢.5.) Cirrus sac; é.v.s., external vesicula seminalis ; /.m., longitudinal muscles ; /.7., lateral nerves ; m.n., median nerve ; 0., ovary; .s., receptaculum seminis; ¢.m., transverse muscles ; v., vagina ; v.e.v., ventral excretory vessel ; v.g., vitelline glands. x I5. Receptaculum and vagina. In segment 16 the vagina is well defined. From the pore it runs as a narrow tube ventral to the cirrus pouch for a short distance, then crosses it posteriorly to reach the dorsal surface. At this point it dilates and runs inwards just below the dorsal transverse muscle fibres, to enter the large receptaculum seminis. The receptaculum seminis 1s roughly club- shaped, with a greatly dilated internal extremity which is bent upon AM.B.del. u. VAY. In. Fic. 3. A. rbodesiensis. Aporal extremity of segment, viewed anteriorly, showing uterus, longitudinal muscles, nerves and excretory vessels. /.m., longitudinal muscles ; /.7., lateral nerves ; m.n., median nerve ; ¢.m., transverse muscles; w., uterus; ©.¢.v., ventral excretory vessel. X 40. 254 itself and almost reaches the ventral transverse muscle fibres. The vagina and receptaculum are always loaded with packets of spermatozoa (figs. 1 and 2). From its median surface there arises a small tube—the fertilisation canal—which runs dorsally, receiving the ducts of the shell gland, vitelline glands and ovary, and finally opens into the uterus (fig. 4). The genital pores are all dextral. Vitelline glands. Appear for the first time about segment 7 and are mature about segment 40. They le ventrally, close to the internal extremity of the receptaculum seminis (fig. 2), and consist of two wings which do not appear to be lobular as in some species of Anoplocephala. Shell gland. As far as could be ascertained, it lies dorsal to the vitelline glands and is almost always obscured by them. Uterus. Visible for the first time about segment 15 as a cell- string running across the segment. In segments 40 to 70 or 80 it is DORSAL u. fe rs. vd od. ~- A.M.B. del. eee VENTRAL Fic. 4. A. rhodesiensis. Diagrammatic representation of fertilisation canal and connecting ducts. f.c., fertilisation canal; o0.d., oviduct; 7.5. receptaculum seminis; w., uterus; 2.d., vitelline duct. X 250. well developed and contains ova; at this point it is a straight wide tube devoid of outgrowths, it does not occupy the whole of the dorso-ventral diameter of the segment, nor does it extend laterally beyond the ventral excretory vessel (fig. 3). Further development beyond this stage was not seen, all the posterior segments being sterile. Eggs. Notwithstanding the fact that about sixty worms were examined, no mature eggs were found. Eggs varying in size from 12 to 25m were often found in the same uterus. The largest eggs 255 seen had apparently three envelopes, the outer measuring about 25, the middle about 20#, and the inner, which was completely filled with the embryo, about 114. These were, however, very rare, and by far the largest number of eggs seen measured about 12s to 15“ in diameter. They contained large oil and yolk globules. All attempts to discover a pyriform body failed. Scattered amongst the eggs in the uterus there occurred large numbers of other cellular structures apparently of a nutritive character (fig. 5). They varied in size, within wide limits, and many seemed to be in process of degeneration. AMB.del. Fic. 5. A. rhodesiensis. Eggs and nutritive cells. ¢., egg; 1.c., nutritive cells, x 730. DIAGNOSIS. The following are the chief diagnostic characters : — 1. Presence of lappets. 2. Its large size and numerous segments. 3. The enormous development of the water vascular system and longitudinal musculature. 4. The large number of sterile segments. Abildgaard, in 1789, described a cestode from a horse, to which he gave the name 7aenia magna; this worm had no lappets behind the head. Zeder, in 1800, also described an equine tapeworm, which had no lappets behind the head, to which he gave the name T. plicata. Rudolphi, in 1808, referring to 7. zedrae, Sander, collected from a zebra, placed it amongst the species dubiae, and stated that it had affinities with 7. plica¢a. Presumably, therefore, 256 T. zebrae of Sander did not possess lappets behind the head. Later, Cobbold expressed the opinion that 7. glzcata was a synonym of 7. magna, Abildgaard. The genus Axoflocephala was established by E. Blanchard in 1848. Collin (1891) gave a general description of a worm collected from an African zebra which resembled A. Ze7folzata in possessing lappets behind the head. He named the species 7aenza zebrae. Gough (1908) states that A. magna (Zeder), var. pediculata, Railliet, were found by him in the horse, donkey and zebra, and that this species is not so rare in S. Africa as elsewhere. He points out that as Collin’s worm possessed lappets, it must be distinct from IT. zebrae, Sander, and is, therefore, a species without a name. Fiihrmann (1910) gave a careful description of Collin’s species of T. zebrae from material obtained by the Kilimandjaro expedition, but did not deal with the synonymy, nor did he refer to Gough’s paper. Fiihrmann has been kind enough to send us a specimen of his A. zebvae, which we have examined, and which is undoubtedly the same species as the one described by us. It should be noted, however, that Fiihrmann in his description fails to mention the interesting fact that the greater part of the worm is sterile (at least the posterior half), and furthermore, that he describes and figures ripe eggs furnished with a pyriform apparatus. These have never been seen by us, notwithstanding the fact that we have examined, minutely, the uterus from the ripest segments of over twenty of our specimens and also that of the specimen Fiihrmann was good enough to send us. Douthitt, in 1915, stated that A. plicata (Zeder) and A. zebrae are synonyms of A. magna (Abildgaard), and draws no distinction between Sander’s 7. zebvae and Collin’s 7. zebrae. He was apparently unaware of Fiihrmann’s paper, as he makes no reference to it. It is clear that the worm which Collin named 7. zebrae differs from Sander’s 7. zebvae, which is apparently identical with T. magna, Abildgaard, in that Collin’s species is lappeted, whereas Sander’s is not. TZ. zebrae, therefore, is a synonym of 7. magna, and Collin’s worm is, as Gough says, a species without aname. We propose to designate it A. rhodeszensis. 257 II. ANOPLOCEPHALA PERFOLIATA (Goeze, 1782), Blanchard, 1848. The museum of this School contains the following collection of lappeted Axoplocephala from horses and mules :— Horse A. Three specimens from Chesterfield, collected by A. W. Noel Pillers, December, 1909. Horse B. Eight specimens from Chesterfield, collected by A. W. Noel Pillers, July, 1910. Horse C. Four specimens from Sheffield, collected by A. W. Noel Pillers. No date. Horse D. Two specimens from Manititlan, Mexico. No further informa- tion. Mute E. Ten specimens from Argentine, collected in 1917. As a result of preliminary examination of the anatomy of specimens from the above sources, we reached the conclusion that a number of different species were represented. On further examina- tion, however, we were impressed with the fact that great variations occurred not only in worms from the same source, but even in different segments of the same specimen. This led us to the conclusion that the differences which at first we considered to be of specific value were of inconstant occurrence, and consequently that A. perfoliata (Goeze, 1782) is a species which exhibits considerable variation. In this paper we propose to re-describe the worm, drawing attention to the variations which may occur. EXTERNAL ANATOMY. The worms from the different sources were approximately the same size, varying from about 30 mm. to 45 mm. in length, except in the case of specimens from Horse B, which were fixed in a very extended condition and were of a gelatinous consistency. These were considerably longer, varying from 44 mm. to 70 mm. Asa rule, the worms attain a maximum breadth of about 12 mm. Head. This is prominent and almost cubical in shape; the length (2°5 mm.) is nearly equal to the breadth (3 mm.). In the specimens from Horse A the head was, however, distinctly broader (2°75 mm.) than long (1°5 mm.). The suckers and lappets resemble closely those of A. rhodesiensts. 258 Segments. The number of segments in an adult worm varies from about ninety to one hundred and‘thirty. The shape of the worm varies enormously, as will be seen in the photographs. Except that the worm is much less massive and consists of fewer segments than A. rhodesiensis, it exhibits no constant external difference from the latter species (Plate XVII). Fic. 6. A. perfoliata. Ventral view of anterior portion of a worm from Horse B; cleared in carbolic acid. c.s., cirrus sac; /., lappets; m.n., median nerve; 0.) ovary; 7.S.. receptaculum seminis; ¢., testes; u., uterus; v.e.v., ventral excretory vessel; v.g., vitelline glands. X 6. INTERNAL ANATOMY. Muscular system. As in A. rhodesiensis, except that the iongi- tudinal fibres are not so strongly developed. 5 Nervous system. “The number of longitudinal nerves on each side varied. As a rule, there were three on each side, but in specimens from Horse B only the. median nerve was constantly present, although the dorsal and ventral nerves were also seen in some segments. This variation was observed in the different 259 segments of the same worm, with the result that some segments exhibited only one nerve on each side, whilst in others three nerves were found on both sides, and in still others three nerves on one side and one on the other. Excretory system. This differed from that of A. rhodesiensis only in that the vessels were not so large. Genitalia. The segments become increasingly ripe towards the posterior extremity, and only a few sterile segments were found scattered here and there. This is in striking contrast to the long chain of sterile segments forming the posterior half of A. rhodeszensis. Moreover, examination of the posterior extremities of worms from various horses left no reason to doubt that segments are shed. DORSAL VENTRAL Fic. 7. A. perfoliata. Segment, viewed posteriorly, showing male genitalia. c.s., cirrus sac ; é.v.5., external yesicula seminalis; 7.v.s., internal vesicula seminalis; /.m., longitudinal muscles ; i.n., lateral nerves; m.n., median nerve; 0., ovary; 7.s., receptaculum seminis; ¢., testes; ¢.m., transverse muscles; v., vagina; v.e.v., ventral excretory vessel. X 20. Testes. These first appear about segment 12 and disappear about segment 30. They attain their maximum development between about segments 17 and 22. In all the specimens examined, excepting those from Horse B, the fully developed testes occupied the whole parenchyma between the aporal excretory vessel and the cirrus pouch (fig. 7). In Horse B, whilst this state of things was occasionally found, the testes, as a rule, were fewer in number and more limited in distribution, being congregated mainly in the dorsal 260 portion of the parenchyma; they did not extend laterally nearly so far as the aporal water vessel. There appears to be no doubt that in old worms the testes degenerate and entirely disappear. No trace of testes were found in the worms from Horse A. Vas deferens. The appearance of the internal and external seminal vesicles varied considerably, and they did not exhibit any constant relationship one to the other. The outer seminal vesicle was usually ventral ‘to the inner seminal vesicle, but this wads not invariably the case; sometimes the former lay directly mternal to the latter, or even dorsal to it. Ovary. This first appears about segment 25 and disappears about segment 45. It attains its maximum development about segment 37. The poral wing has a diameter of about half that of the aporal wing. In structure it resembles that of A. rhodesiensis DORSAL VENTRAL Fic. 8. A. perfoliata. Segment, viewed posteriorly, showing female genitalia. e.v.s., external vesicula seminalis ; 7.v.s., internal vesicula seminalis ; /.m., longitudinal muscles ; /.7., lateral nerves > m.n., median nerve ; 0., ovary; 7.5., receptaculum seminis ; t.m., transverse muscles; v., vagina ; v.e.v., ventral excretory vessel; v.g., vitelline glands. x 20. (fig. 8). As in the case of the testes, the ovaries had entirely disappeared in worms from Horse A. Receptaculum and vagina. ‘The vagina appears early and is well defined in about segment 8. These structures resemble in all respects those of A. shodesiensis. Vitelline and shell glands. Similar to those of A. rhodesiensis. Uzerus. The uterus appears very early, about segment 12, as a delicate cell-string running across the segment. It gradually 261 enlarges and attains its full development only in the last few segments, where it is a wide tube completely filling the medullary parenchyma, but only occasionally crossing the ventral excretory vessels. In the fully developed uterus there are numerous anterior and posterior.outpocketings, so that in anterior or posterior views of whole segments the uterus appears to be composed of a number of separate compartments (fig. 9). Sections, however, showed clearly Fic. 9. A. perfoliata. Segment, viewed posteriorly, showing fully developed uterus. ., egg ; 9 Ap gment, P } g fully P » BB; I.m., longitudinal muscles ; t.m., transverse muscles; u., uterus. X 40. that the organ consisted of a central cavity with numerous anterior and pesterior bulges between the dorso-ventral muscle fibres (fig. 10). To a certain extent there are bulges on the dorsal and ventral surfaces of the uterus, but they are not so prominent as those on the anterior and posterior surfaces. The degree of outpocketing varied in A.M.B.del. Fic. 10. A. perfoliata. Horizontal section of segment showing fully developed uterus. X 40. different worms, sometimes being well marked and in other cases less obvious. Of the worms examined by us, the outpocketing was most marked in those from Mule E, and least marked in those from Horse B. Occasionally a well-developed uterus was seen which contained few or no eggs and a mass of debris. Eggs. Fully mature eggs were found only in the last few segments. The diameter of the outer envelope was about 80”, and that of the embryo about 16”, whilst the length of the horns of the pyriform apparatus is about 184. Yolk granules about 7m in 262 diameter occurred plentifully (fg. 11). As the egg matures, the middle envelope gradually shrinks, and its measurement is therefore of no value. In quite ripe eggs the horns of the pyriform apparatus A.M.B.dek Fic. 11. A. perfoliata. Eggs showing pyriform apparatus. X 360. are prolonged into very long, slender filaments which eventually unite with one another. DIAGNOs!Is. Fihrmann, as a result of a comparison between his own observations on the lappeted Axoplocephala from the zebra (called by him A. zeérae), and Kahane’s description of A. ferfolzata, gives the following points of difference between the two worms. Cirrus pouch. In A. zebrae it extends beyond the longitudinal nerve and ventral water vessel, whilst in A. perfoliata it scarcely reaches the water vessel. Vesicula seminalis. In A. zebrae, it is dorsal or ventral to the cirrus pouch, and in A. ferfolzata it is posterior. Ovary. Much more strikingly asymetrical in A. zebrvae than in A. perfoliata. We have carefully examined these points, and have reached the conclusion that they have no specific value. In the specimens of A. perfoliata examined by us, the cirrus pouch extended over the longitudinal water vessel and nerve to the edge of the segment, just as it does in the worm from the zebra. The relative position of the outer seminal vesicle and the cirrus pouch is inconstant, and finally 263 the aporal wing of the ovary is about twice the size of the poral wing in both worms. In our experience, the only constant points of difference between the lappeted Axofplocephala of the zebra and _ horse, viz., A. rhodesiensis and A. perfoliata, are as follows :— (1) A. rhodestensis is much more massive than, and has almost twice as many segments as, A. ferfolzata. (2) The posterior half of A. rhodesiensis is entirely sterile, whereas in A. perfoliata the segments become increasingly ripe up to the posterior extremity of the worm. REFERENCES Bepparp, F. E, (1911). Contributions to the Anatomy and Systematic Arrangement of the Cestoidea. I: On Some Mammalian Cestoidea. Proc. Zool. Soc., London. ——— (1911). Contributions to the Anatomy and Systematic Arrangement of the Cestoidea. II: On Two New Genera of Cestodes from Mammals. Proc. Zool. Soc., London. (1912). Contributions to the Anatomy and Systematic Arrangement of the Cestoidea. IV: Ona species of Inermicapsifer from the Hyrax, and on the Genera Zschokkeella, Thysanotaenia, and Hyracotaenia. Proc. Zool. Soc., London. Biscnorr, C. R. (1912). Cestoden aus Hyrax. Zool. Anz., 39. Brancuarp, R. (1891). Notices helminthologiques (deuxiéme séries). Mém. Soc. Zool., France, 4. Cottin, A. (1891). Parasiten aus dem Darm des Zebra. Sitz. des Ges. Nat. Fr. xu Berlin. Deiner, E. (1912). Anatomia der Anoplocephala latissima (nom. nov.). Arb. Zool. Inst., Wien., 19. Dournirr, H. (1915). Studies on the cestode family Anoplocephalidae. Illinois Biological Monographs, University of Illinois. Fiiurmann, O. (1902). Die Anoplocephaliden der Vogel. Centralbl. f. Bakt., 1, Abt., Orig. 32- (1910). Vermes. Wissenschaftliche Ergebnisse der Schwedischen Zool. Exp. nach dem Kiliamndjaro dem Meru, 1905-6. Band 3, Abt. 15-22, Stockholm. Goucu, H. L. (1908). Notes on Some South African Parasites. South African Association for the Advancement of Science, Grahamstown. Hatt, M. C., and Hoskins, P. H. (1918). The occurrence of Tapeworms, Anoplocephala spp-, of the Horse in the United States. Cornell Veterinarian, October. Kanane, Z. (1880). Anatomie von Taenia perfoliata, Goeze, als Beitrag zur Anatomia der Cestoden. Zettschr. f. wiss. Zool., 34. Lerpy, J. (1855). Notices on some Tapeworms. Proc. Acad. Nat. Sc., Philad., 7. Neveu-Lemaire, A. (1912). Parasitologie des Animaux Domestiques, Paris. von Linsrow, O. (1gor). Helminthen von den Ufern des Nyassa-Sees—ein Beitrag zur Helminthen-Fauna von Siid-Afrika. ‘fen. Zeitschr. f. Naturw., 35- MacCattum, G. A., and MacCatium, W. G. (1912). On the Structure of I. gigantea. Zoolog. Fabrb. Syst., 32. Neumann's Parasites (1905). Macqueen’s translation, 2nd edition, London. Rartuier, A. (1893). Traité de Zoologie médicale et agricole, 2nd ed., Paris. Henry, A., et Baucue, A. (1914). Sur les Helminthes de L’Eléphant d’Asie. Bull. Soc. Path. Exot., Tome VII, Nos. 1, 2, and 3. Ransom, B. H. (1909). Taenoid Cestodes of North American Birds. Bull. U.S. Nat. Mus., 69. Suiprey A. E. (1900). A Description of the Entozoa, collected by Dr. Willey during his sojourn in the Western Pacific, (Jn Willey, Zool. Results, Part 5, Cambridge.) Stires, C. W. (1895). Notes on Parasites, No. 38. Preliminary Note to ‘A revision of the adult leporine cestodes.’ Vet. Mag. (1896). A Revision of adult tapeworms of hares and rabbits. Proc. U.S. Nat. Mus. and Hassatr, A. (1902-1912). Index-catalogue of Medical and Veterinary Zoology. Author’s Index. Bur. An. Ind. Bull., 39. (1912). Index-catalogue of Medical and Veterinary Zoology. Subjects: Cestoda and Cestodaria. Hyg. Lab. Bull., 85. Zscuoxxe, F. (1888). Recherches sur la structure anatomique et histologique des Cestodes, Genéve. 264 EXPLANATION OF PLATE XVII Fig. 1. Photcgraph of Axoplocephala rhodesiensis showing bots attached. Natural size. Fig. 2. Photograph of Axofplocephala perfoliata from Mule E, Horse B and Horse C, and of Anofplocephala rhodesiensis frem a Zebra. (In order from left to right.) Natural size. Annals Trop. Med. & Parasitol., Vol. XV PLATE Xp Brees 265 THE FEEDING HABITS OF STEGOMYIA CALOPUS, MeicEn BY R. MONTGOMERY GORDON AND C. J. YOUNG From the Laboratory of the Liverpool School of Tropical Medicine, Mandaos (Received for publication 28 July, 1921) It has been stated by Marchoux and Simond (1906) that Stegomyia calopus, under normal conditions, that is, while at liberty, ceases to bite man during the day after the first six or eight days of its existence in the perfect state, but that after the eighth day it is sometimes observed to bite towards 6 p.m. before nightfall. They also state that yellow fever is not contracted during the day, but only ‘a la chute du jour ou pendant la nuit.’ ‘Nous avons en effet constaté expérimentalement qu’a la période de sa vie ot il possede le pouvoir infectant, le Stegomyia fasciata en liberté ne cherche pas a piquer homme entre 7 heures du matin et 5 h. 4 du soir. La transmission est donc nocturne.’ This they held to account for the fact that the inhabitants of Petropolis when visiting Rio only during the day did not contract yellow fever. In their experiments, however, the mosquitoes were confined in a screened room. Seidelin and Connal (1914) and Macfie (1915-16) showed that S. calopus would bite at any hour irréspective of age, when in captivity. According to Marchoux and Simond (1906) the minimum interval between the date on which S. calopus acquires the infection and that on which it becomes infective is twelve days. The following experiments were undertaken with a view to investigating the feeding habits under as natural conditions as possible. They were carried out in Manaos. All the mosquitoes were hatched in the laboratory and kept in wire gauze cages, males being always present. Each female was removed from the stock cage in a glass tube and allowed to feed to repletion on one of us during the hours of daylight. After feeding they were ‘marked’ by amputating the hindlegs through the tibiae, 266 but in a few instances some part of the femur was accidentally removed. This method of ‘marking’ 1s considered sufficiently distinctive as wild specimens of Stegomyia, observed by us at other times, have never shown this mutilation. After feeding they were kept for not less than fourteen days, sugar being available as food, and during the last two or three days of this period were again given an opportunity to bite, also during daylight. They were then released under the conditions described below and during the four succeeding days, in the place where they had been released, the mosquitoes biting us were observed for one hour each during the day and night. The observations at night were made by electric light. No unmarked females were released along with the ‘ marked.’ Three experiments were carried out. Experiment I. Date: 23rd March to 16th April, 1921. Sixty female Stegomyia were fed a first time and ‘marked’ between 23rd and 28th March. Of these thirty-three survived, and were offered a second feed on the 11th and 12th of April; thirty fed and three refused to feed. Two escaped after feeding. The remaining thirty-one females along with twelve males were released at 7 p.m. on the 14th April in a first-floor bedroom of about 14 ft. by 12 ft. by 10 ft. This room was in use, the occupant using a mosquito net at night; it had two unscreened windows which were ccnstantly open, and the mosquitoes also had free access to other parts of the house. A glass jar containing water was placed in the rcom, but no eggs were deposited in it up to the 23rd April. ‘Marked’ mosquitoes were released after feeding. Observations were made as follows :— | | Date ... ...| April i 15 | 16 | 17 18 | 12.30 | 8.30 | 12.30 | 8.30 | 12.30 | 8.30 12.30 8.30 Time ... ..| P.M. “| to to | to to to to to to | 1.30 9-30 | 1.30 9-30 1.30 9.30 1.30 | 9.30 | | “Marked’...) 3 ° 2 ° 0 ° 0 ° S. calopus biting Unmarked...) 3 he |e 2) ° | ° ° ° ° | NoTE.—In addition, one of us was bitten by a ‘marked’ mosquito at 7.30 a.m. on the 15th. 267 Experiment II. Date: 5th to 29th April, 1921. Sixty female Stegomyzia were fed a first time and ‘marked’ between the 5th and 11th April. Of these twenty-three survived. “They were offered a second feed on the 23rd and 24th April; twenty-one fed and two refused. They were released with about twelve males at 5 p-m. on the 25th in a first-floor bedroom in a different part of the town frem-Experiment I. This room was about 20 ft. by 14 ft. by 12 ft. in size, was in use, the occupant using a mosquito net at night, had four large windows constantly open, and the mosquitoes had free access to other parts of the house. Observations were made as follows :— Date ... .--| April = 25 26 27 28 29 Time ... ...| P.M. --.| 8—g | 12—1 | 8—g | 12—1 | 8—g | 12—f | 8—g | 12—1 ( ‘Marked’...| 0 0 ° OA.) ac 0 ° 0 S. calopus | biting... a et Unmarked...| 0 ° 2 ° ° ° ° ° } Experiment III]. Date: 1st May to 3rd June, 1921. Seventy female Stegomyta were fed a first time and ‘marked’ between ist and 14th May. Of these fifty survived. Offered second feed on 28th, 29th and 30th May; forty-three fed and seven refused. These fifty were released along with about twelve males at 3 p.m. on the 30th May in a ground-floor room behind the laboratory, about 17 ft. by 1oft. by 15 ft. in size and opening directly to the outside. This room had one window, the upper part of which was imperfectly screened and the lower part had wooden slats, the apertures between these allowing free passage to mosquitoes. The door was open all day and for one hour at night when observations were made. Otherwise the room was not used at night. A jar of water was placed in the room, and some eggs were found on 31st May. .‘ Marked’ mosquitoes were released after feeding. 268 Observations were made as follows :— Date ... ---| May =| 30 31 June 1 2 3 ‘ | : 8.30 | 8.30 8.30 8.30 Time ... ..-| P.M. eel ta: I—z | to I—2 to I—2 to I—2 | 9-30 | | 9-30 9-32 9:30 ‘ Marked ’... 3 7 c} 2 ° 1 ° 0 S. calopus biting | —— Unmarked... I 4 4 I 3 ° ° 5 Note.—Each of us was bitten in the laboratory by a ‘marked’ Stegomyia on the 7th June, one at 9 a.m. and the other at 4 p.m. SUMMARY Experiment I. Thirty-one ‘marked’ female S/egomyia were released at night not less than fourteen days after their first blood meal. During the succeeding four days, five ‘marked’ and three unmarked fed during daylight. None were observed to feed at night. Experiment II. With the exception of two unmarked Stegomyia, none were observed to feed. Experiment III. Fifty ‘marked’ female Stegomyia were released during daylight not less than fourteen days after their first blood meal. During the succeeding four days, ten ‘marked’ and ten unmarked fed during daylight, and six ‘marked’ and eight unmarked fed during the night. CONCLUSION Stegomyia calopus females will bite either by day or night, over fourteen days after their first blood meal, while under no artificial restraint and having opportunities of selecting day or night for feeding. REFERENCES Macrir, J. W. S. (1915-16). Bulletin of Entomological Research, Vol. V1. Marcuovx and Simonp (1906). Annales de I’ Institut Pasteur, Vol. XX. SerpeLtn and Connat (1914). Yellow Fever Bulletin, Vol. II, No. 3. 269 NOTE ON A CASE OF LEPROSY BY WARRINGTON YORKE AND S, ADLER (Received for publication 2 August, 1921) PLATE XVIII Native of Hong Kong, age 45. Came to the outpatient department of Tropical School, 29th July, 1921, complaining of feeling ill, ulceration of face and generalised skin eruptions. PRESENT CONDITION. The appearance of the face at once suggested leprosy. Both alae masz were swollen and ulcerated, and there were fairly well developed nodules on the chin, forehead and cheeks. There was an extensive raised, ham coloured, rash on the arms, forearms, trunk and legs. The affected areas of skin were not anaesthetic except for small patches on the inner side of the left leg and foot and on the outer side of the right leg. No thickening of. the superficial nerve trunks was observed. Scrapings from the skin lesions and from the alae nasi showed very numerous lepra bacilli. History. The most interesting feature of this case is its history. The patient arrived in England in 1912 in good health, and has been employed until a short time ago as a laundryman in Cardiff. It was not until 1916—four years after coming to England—that he first noticed small spots on the left side of the face. No further change was observed until twelve months ago, when the ham- coloured eruption appeared on the trunk and upper and lower limbs. Five months ago he noticed for the first time the swelling of the alae nas. There is no evidence that the case had been diagnosed before we saw him: the patient and his friends were quite unaware of the fact that it was leprosy. 270 EXPLANATION OF PLATE XVIII Photograph of a case of leprosy showing swelling and ulceration of alae xasi and nodules on various parts of face. Annals Trop. Med. & Parasitol., Vol. XV PLATE XVill C, Tinling & Co., Ltd., Imp. 271 NOTES ON SOME FUNGAL INFECTIONS IN WEST AFRICA BY Jj; WW. 2. | MACE IE (Received for publication 24 August, 1921) CONTENTS PAGE I. A Funeous or tHe Genus Monilia RecovereD FROM A PaTIENT SUFFERING FROM DysENTERIC DIARRHOEA ... wee aa2 ae a oes cet napl t L2R2 II. Two Funer or tHe Genus Monilia RecoverED FROM AFRICANS SUFFERING FROM A Pecutrar Form or DIARRHOEA ste tee oe si ae sete 25 Ill. A Note on a Cask oF Oromycosis ... tee a ns Are ras ree a) IV. Yonstttrar Nocarpiomycosis ... bx rh aD ae ee #.; anu: py Fungal infections are abundant in West Africa, but hitherto comparatively little attention has been devoted to them. Brief notes are given in this paper on a few cases which have recently come under our notice at Accra in the Gold Coast. The part played by the fungi of the Genus J/onzlza in the diseases from which the patients were suffering is obscure. Such fungi are undoubtedly abundant in the tropics, and contamination with them must be guarded against; it is also true that they are not infrequently present in small numbers in the faeces of apparently healthy individuals.. Their occurrence in great quantities 1s, however, a different matter. No evidence is brought forward in support of the view that the species of Monilia described in this paper were the sole cause of the dysentery and diarrhoea with which they were associated, indeed there was evidence that the cases referred to in Section II were due primarily to a dietetic deficiency, but it is maintained that they were the cause of some of the symptoms. For this reason, and because the intestinal disturbances which permit of their multiplication may be of profound significance, their recognition and investigation is of importance. 272 I. A FUNGUS OF THE GENUS MONILIA RECOVERED FROM A PATIENT SUFFERING FROM DYSENTERIC DIARRHOEA Towards the end of the year 1920 there occurred at Accra a considerable number of cases of dysenteric diarrhoea, the cause of which was obscure. From one such case, a European man about forty years of age, the Mozzlza was isolated, of which the following is a Short description. The sample of faeces from which the fungus was recovered contained much blood and mucous, but no amoebae and no other parasite to which a pathogenic réle could be assigned ; it contained, however, a considerable number of yeast-like cells. From the same case, a week later, Dr. J. F. Corson isolated a bacillus which gave the reactions of Morgan’s bacillus. Several species of the Genus Moznzilia have been isolated from human faeces in various parts of the world, but especially in the tropics, and some of them have been regarded as being the cause, either directly or indirectly, of intestinal disease. Similar fungi are frequently found in small numbers in specimens of faeces in West Africa, and in diarrhoeic stools are sometimes very numerous. With regard to the species isolated from this case of dysenteric diarrhoea at Accra it is not possible to say if it was pathogenic, but it may be noted that it closely resembled d/. ¢vopzcalzs, one of the species which has been found to be present in sprue (Castellani, 1920). The Organism. The Monilia found in this case was isolated in a culture of the faeces on neutral-red lactose bile-salt agar. It was Gram-positive and not acid fast. It grew well on most solid media, and on glucose agar produced within twenty-four hours an abundant creamy-white growth. Under anaerobic conditions growth was less abundant and less rapid. It grew freely both at 37°C. and at the temperature of the laboratory (about 26° C.). Gelatin and blood serum were neither liquefied nor stained. In broth and peptone water a whitish deposit was thrown down, whilst the media themselves remained clear; in both a surface pellicle was formed. It produced an abundant white growth on potato, which later developed a white efflorescence; the medium was not stained. On solid media the growth was mainly composed of yeast- like cells, but a few branched septate hyphae were usually present; in fluid media the hyphae sometimes predominated. 273 Its qualitative bio-chemical reactions may be tabulated as follows, the symbols representing:—A = acidity; G = gas; s = slight; O = neither acid nor gas. - Arabinose te oe O Inulin O Rhamnose (isodulcite) ... O Amygdalin O Galactose te: ase -AGs Helicin ... O Glucose ... “te ws) AG Phlorrhizin O Laevulose 2h ey. \ eC; Salicin O Mannose... bi ae O Glycerol O Lactose ... oe aa O Erythrol O Maltose ... Ba un JAG Adonitol O Saccharose Ra . AGs Dulcitol ... O Raffinose... bs 20 O Inosite O Amylum be. See O Mannitol O Dextrin ... aM be O Sorbitol ... O Glycogen = ead O If the cultures were kept, the acidity produced in the five sugary media indicated tended to be superceded by alkalinity ; this change began to occur early, usually in less than a week. No change was produced in Litmus milk at first, but after a week or ten days a shght alkalinity was sometimes noticed; no clot was formed, and the medium was neither decolourized nor peptonized. Indol was not produced in peptone water. As gas was produced in Glucose, Laevulose, Maltose, Galactose, and Saccharose the organism comes into the fifth group of species of Monilia, the Tropicalis group, according to the ‘classification of Castellani and Chalmers (1919). The more important bio-chemical reactions of the species in this group are shown in Table I. It will be noted that the species here described differs in its reactions in Mannitol and Dextrin from M. enterica, M. insolita, M. para- tropicalis, and M. pulmonalis. From M. faecalis it may be distinguished by the fact that it does not decolourize Litmus milk, and does not stain blood serum brown; and from M. metatropicalis by the fact that it does not clot milk. As regards M. nivea, the fact that neither acid nor gas is produced in Raffinose is a point of distinction, and it may be noted also that our species produces only a slight amount of gas in Galactose and forms a pellicle on the surface of broth, reactions which are not found in the case of M. nivea. Our organism closely resembles M. tropicalis, the greatest divergence being in the reaction in milk. It is said that by M. ¢ropicalis ‘Litmus milk is generally rendered acid but is not 274 clotted,’ a statement which would seem to imply that the production of acid is not constant nor characteristic. species appear to be identical, excepting that in the case of our organism a pellicle is formed in broth. Taste I. If this is so, the two Species of the Genus Monilia belonging to the Tropicalis group. a m= o & 2. “4 FS > o & Species of Monilia g 2 3 2 2 5s bs = 8 g is] 3 = zs S 3 fo] x ne re} Se = S S “s S = 2 GH = 4 2) 4 = (e) a a fa) % < M. enterica O/Alk | AG | AG | AG | AG | AG | As As oO oO M. faecalis A/DPs | AG | AG | AG | AGs| AGs| O 0) na M. insolita As/Alk |} AG | AG |} AG | AG | AG] As fe) oO M. metatropicalis AC AG | AG | AG | AG |} AG oO oO 10) M. nivea O/Alk | AG | AG | AG | AG |} AGs| O AG oO M. paratropicalis As/Alk | AG | AG | AG | AG | AG oO Avs O oO M. pulmonalis -|O/AlkD | AG | AG | AG | AGs | AG | Avs oO A | AGs M. tropicalis A AG | AG | AG | AGs} AGs|] O oO oO oO Accra case : sputumand pleural | O/Alk | AG | AGs | AGs | AG | AG oO O 10) cavity (M. accraensis) t . Accra case: dysenteric faeces ...| O/Alk | AG | AG | AG | AGs | AGs| O oO oO CEP A = acidity; Alk = alkalinity ; C = clot (milk), clear (broth) ; CTP = clear, then pellicle ; D = decolourized ; G= gas; O = neither acid nor gas; P = peptonized; s = slight; vs = very slight. Finally it may be noted that this organism closely resembles a species of Menzlza (M. accraensis) recently found by us in the sputum and pleural cavity of a patient suffering from tuberculosis at Accra (1921). The chief points of distinction are that the intestinal species produces only a slight amount of gas in Galactose and Saccharose, and forms a pellicle on broth; whereas the pulmonary species produces much gas in Galactose and Saccharose but only little in Laevulose and Maltose, and does not form a pellicle on broth. SUMMARY From the faeces of a European with dysenteric symptoms a fungus of the Genus Monzlia was isolated. This organism belongs 275 to the T7vopicalis group, and appears to resemble most closely M. tropicalis (Castellani, 1909). REFERENCES Casveiant, A. (1920). Milroy Lectures. ‘fourn. Trop. Med. and Hyg., XXIII, p. 120. and Cuarmers, A. J. (1919). Manual of Tropical Medicine. Bailliere, Tindall and Cox, London. pp. 1082-1083, and p. 1086. Macriz, J. W. S., and Incram, A. (1921). Bronchomoniliasis Complicating Pulmonary Tuberculosis in a Native of the Gold Coast, West Africa. -dnnals Trop. Med. & Parasitol., XV, pp. 53-58. Il. TWO FUNGI OF THE GENUS MONILIA RECOVERED FROM AFRICANS SUFFERING FROM A PECULIAR FORM OF DIARRHOEA There occurs among the natives at Accra a well defined form of persistent diarrhoea which has been’ considered by some of the medical practitioners in the Gold Coast to be akin to sprue. The stools are bulky, frothy, and generally canary-yellow coloured; the tongue is red, fissured, and eroded; and the patients are wasted— a condition which is often masked by oedemas of the limbs and by ascites. The faeces in such cases usually teem with yeast-like cells which on cultivation are found to be stages of fungi of the Genus Monilia. The following are short descriptions of the organisms isolated from two such cases. FIRST CASE. The patient, a native man about twenty-seven years of age, was suffering from an obstinate diarrhoea, and was weak and wasted. His tongue was red, irregularly fissured, indented by the teeth, and partially eroded ; the throat also was red. The faeces were canary- yellow coloured and frothy ; yeast-like cells were extremely abundant in them, and Entamoeba coli and Blastocystis enterocola were also present. The Organism. The Monilia in this case was isolated from a culture of the faeces on neutral-red lactose bile-salt agar. It was Gram-positive and not acid fast. It grew well on most solid media, and on Glucose agar produced within twenty-four hours an abundant white growth, very fluid in consistence, and with a dull pellicle-like surface. The yeast-like cells, which composed the greater part of the growth, were oval and somewhat elongated; the average 276 measurements of ten cells were, length 4°54, breadth 16”. Under anaerobic conditions growth was less abundant and less rapid. Gelatin and blood serum were neither liquefied nor stained. In broth and peptone water a whitish deposit was thrown down, and the medium remained clear; in broth a surface pellicle was formed. On potato a dull white growth was developed which later showed a white efflorescence; the medium was not stained. On solid media the growth was mainly composed of yeast-like cells, but a few branched, septate hyphae were also present; in fluid media hyphae sometimes predominated. The qualitative bio-chemical reactions of this M/onzlia may be tabulated as follows :— Arabinose : O Tnulin O Rhamnose (isodulcite) ... O Amygdalin O Galactose 3 es O Helicin ... As Glucose ... we a ING Phlorrhizin Oo Laevulose OS soe en AG Salicin O Mannose... O Glycerol O Lactose ... ae 366 O Erythrol O Maltose ... ae +. ot AGs: Adonitol O Saccharose a 121 4 AG Dulcitol ... O Raffinose... eae con, AGS Inosite O Amylum ee Le O Mannitol Oo Dextrin ... sh 8 TAG Sorbitol ... Oo Glycogen so 56 O The symbols representing : A = acidity ; G= gas; s= slight ; O = neither acid nor gas. In the media in which acidity was produced the reaction tended after a few days to be succeeded by alkalinity. No change was produced in Litmus milk. Indol was not produced in peptone water. As gas was produced in Dextrin as well as in other media the organism comes into the ninth group of species of Monzlia, the Pseudolondinensis group, according to the classification of Castellani and Chalmers (1919). Two species belong to this group, namely, M. pseudolondinensis, Cast., and M. pseudolondinoides, Cast., which appear to differ only in their actions on Litmus milk. The more important bio-chemical reactions of these species and of the organism just described are given in Table II; it will be noted that 277 ; the latter species differs from the other two in its actions on Galactose, Saccharose, and Raffnose. These differences are important, and since the classification of these fungi is at present Taste II. Species of the Genus Monzilia belonging to the Pseudolondinensis group. 5 | swe oe Species of Monilia 2 | 3 g g e 2 / = Bp) Sl Eels |e] €] & ea) = Gi i is) aay as | Oo la ioe | A M. pseudolondinensis, Cast. ap 51410 AG | AG | AG | AG o | ac M. pseudolondinoides, Cast. oe -.| AC | AG | AG | AG | AG oO O | AG M. africana, sp.n. w= | «se eef O.| AG | AG | AGs| O | AG | AGs | AG A=acid; G= gas; s =slight; O = neither acid nor gas; C = clot. based on such bio-chemical reactions, the organism described here must be regarded as a new species. For it, therefore, the name Monilia africana is proposed. SECOND CASE. The patient, an adult native man, a Moshi, was suffering from persistent diarrhoea accompanied by ascites and oedema of the face, especially the left parotid area, the lumbar region, and the penis. There was no oedema of the legs. The faeces were semi-fluid, pale canary-yellow coloured, and frothy; they contained very numerous yeast-like cells. The Organism. The Monilia in this case was isolated from a culture of the faeces on neutral-red lactose bile-salt agar. The colonies of yeast-like cells were rather slow in appearing, being detected first after forty-eight hours’ incubation. It was Gram- positive and not acid fast. It grew well on most solid media, spreading as a thin film, and on Glucose agar produced within twenty-four hours an abundant white growth. The yeast-like cells which composed the greater part of this growth were, as in the previous case, somewhat elongated. Under anaerobic conditions growth was slower. Gelatin and blood serum were neither liquefied nor stained ; the growth on gelatin was slow. In broth and peptone water a whitish deposit was thrown down, the medium remained 278 clear, and no pellicle was formed. On potato an abundant whitish growth was produced, and the medium was not stained. On solid media the growth was composed mainly of yeast-like cells, but a few branched, septate hyphae were also present; in fluid media hyphae sometimes predominated. The qualitative bio-chemical reactions of this J/onilia may be tabuiated as follows :-— Arabinose gts Sas O Inulin... O Rhamnose (isodulcite) ... O Amygdalin O Galactose A Helicin ... As Glucose ... AGs Phlorrhizin O Laevulose AGs Salicin As Mannose... O Glycerol O Lactose ... O Erythrol O Maltose ... A Adonitol oO Saccharose O Dulcitol ... O Raffinose... O Inosite O Amylum O Mannitol O Dextrin ... O Sorbitol ... O Glycogen O The symbols representing ; A= acidity; G= gas; s= slight ; O = neither acid nor gas. In the media in which acidity was produced the reaction tended after a few days to be succeeded by alkalinity. No change was produced in Litmus milk. Indol was not produced in peptone water. As gas was produced in Glucose and Laevulose only, the organism comes into the second group of species of Mowilia, the Krusei group, according to the classification of Castellani and Chalmers (1919). Two species are included in this group, namely, M. kruser, Cast., and M. parakrusei, Cast., which appear to differ only in their actions on Litmus milk. The more important bio-chemical reactions of these species and of the organism just described are given in Table III; it will be noted that the latter species differs from the other two in its actions on Maltose and Galactose besides in several minor points. These differences, according to the system of classification at present in vogue, are sufficient to justify the erection of a new species, and, therefore, although the number of species of the Genus Mozilia is already embarrassingly large, we propose for this organism the name Mowilia enterocola. 279 Taste III. Species of the Genus Monilia belonging to the Krusei group. —| ‘2 2 2. = Species of Monilia 3 2 s 2 8 5 E rs) > z=) 3 oO = = S Ss z ss) 16) 4 = 1e) n M. krusei, Cast. ... aed Re me oO PAG AG Oo M. parakrusei, Cast. eed nA eh SAG AG AG oO M. enterocola, sp.n. oth oe ne Oo AGys AGs A A=acid; G= gas; s=slight; vs = very slight; O = neither acid nor gas; C = clot. SUMMARY From the faeces of two Africans suffering from a peculiar form of diarrhoea two fungi of the Genus Monilia were isolated, both of which appear to be hitherto undescribed species. The one belongs to the Pseudolondinensis group, the other to the Krusez group of Castellani and Chalmers. The names Monilia africana and Monilia enterocola, respectively, are proposed for these organisms. Ill A NOTE ON A CASE OF OTOMYCOSIS Otomycosis is said to be not uncommon in the tropics. It may be caused by a considerable number of different fungi, and according to Castellani and Chalmers (1919), ‘if they grow superficially, they cause no symptoms; but if they penetrate into the mucous membrane, they give rise to itching, and sometimes to pain.’ The patient whose case is the subject of this note was a European lady who consulted Dr. C. V. Le Fanu at Accra in the Gold Coast Colony, West Africa, on account of pain and irritation in the ear. The symptoms had existed for at least a month, but although they caused no little discomfort, there was no deafness. On examination Dr. Le Fanu found that the external auditory meatus was unusually narrow, but widened out at its inner end so as to form a small chamber immediately external to the drum of the ear. From a patch on the vault of this chamber, close to the drum of the ear, the fungus 280 was growing and hanging down like a veil. It may be said at once that the condition was rapidly relieved, and apparently cured, by an application containing as its principal constituent salicylic acid. The fungus, which grew readily on Sabouraud’s maltose agar and was easily obtained in pure culture on this medium, appeared to belong to the Genus Séerzgmatocystis, Cramer 1859. Two species of this genus have been found in cases of otomycosis, namely, S. antacustica and S. nidulans, but the characters of both, as given in the descriptions to which I have access, differ slightly from those of the fungus isolated at Accra. The exact determination of the species of such a fungus is a matter for a mycologist. I shall, therefore, restrict myself to recording certain characters which may enable those who have made a special study of these fungi to place it more exactly. Cultures. On Sabouraud’s maltose agar the growth of the fungus was rapid and spreading; at first yellowish-white and felt-like, then slightly fluffy with the development of upstanding conidiophores, then speckled with dark brown points when the conidiophores began to develop their spores, and finally dark brown or almost black all over, resembling a mass of soot. On Glucose agar the initial growth at a temperature of 28°C. was almost white, with a somewhat puckered and wrinkled surface, but within twenty-four hours became yellowish, and began to show upstanding, white conidiophores. On the following day the whole surface of the medium was covered by a yellow, wrinkled, felt-like growth, on which were very many conidiophores, some already dark brown in colour. On the third day the whole surface of the medium looked as if it had been thickly dusted with soot. Growth, therefore, was rapid and abundant at 28° C., the temperature of the laboratory ; it was, however, even more rapid at 37°C. In peptone water the fungus grew mainly on the surface, but detached fragments in the fluid produced delicate networks of hyphae which sometimes resembled puff-balls. Dark spores were formed on the conidiophores at the surface. In glucose peptone water acid was produced, but no gas. On blood serum growth was rather slow, the colonies were white, and the black spore- bearing conidiophores did not develop for a long time and were relatively scanty. Eventually the medium was liquefied. In a 281 gelatin stab culture there was no deep growth, and no liquefaction. A white surface growth developed with dark sooty grains, and later, just below the surface, there formed compact cerebriform whitish masses of the fungus. On potato growth was rapid and abundant; at first a yellowish felt-work, finally a sooty mass. In Litmus milk acid and clot were produced. The growth was mainly at the surface, was yellowish, and developed the usual dark brown sooty appearance. Mycology. All the growths showed septate, branched hyphae of varying diameter. The hyphae appeared colourless when seen with a microscope. The lengths of the interspaces were very variable. In fluid media, such as glucose peptone water, chlamydospores were numerous in the surface felt-like growth. The conidiophores were erect. In cultures they were raised about 1 mm. above the level of the medium, but in the ear of the patient they appeared to be considerably longer. They were white on first appearance, but darkened later. The diameter of the stem was variable, but averaged about 14m. The head was almost spherical, slightly broader than long, its diameters averaging 40m and 374 respectively ; it was covered almost completely by sterigmata, only a very narrow zone at the proximal end, at the insertion of the stalk, being free from them. Both primary and secondary sterigmata were present; the primary sterigmata were about 12 long (average of ten), the secondary about 8» long (average of ten). There were usually four secondary sterigmata on each primary sterigma. The spores were dark brown in colour, and spherical in shape; diameter 4p to 5'2y, average 45h. Animal inoculations. Two wild Mus rattus were given intra- peritoneal inoculations of an emulsion of a small fragment of the original culture in normal saline. No ill-effects were observed to follow. A little of the same culture was applied to the external auditory meatus of a pouched rat (Cricetomys gambianus), a sheep, . and a small monkey (Cercopithecus patas) after scarification, but no otomycosis was produced. REFERENCE Castriiant, A., and Cuarmers. A. J. (1919). Manual of Tropical Medicine, Third Edition, p- 2012. London: Bailliere, Tindall and Cox. 282 IV. TONSILLAR NOCARDIOMYCOSIS One case of tonsillar nocardiomycosis has been met with at Accra. The patient, a European man, about thirty-two years of age, showed a number of small white concretions in crypts of both tonsils. The throat was not inflamed, and there was neither pain nor discomfort ; but the presence of the concretions had been detected by the patient at least a month previously, and he was worried about them as they did not show any signs of disappearing. Scrapings from one of the white patches consisted mainly of fungal hyphae in short lengths. They were about 1 in diameter, branched, and either Gram-positive or composed of Gram-positive coccoid bodies and rods connected together by Gram-negative strands. Inoculations made on agar and glucose agar were unsuccessful. Further particulars with regard to the fungus cannot, therefore, be given. This condition is referred to by Castellani and Chalmers (1919) in their Manual of Tropical Medicine. The above case is recorded here merely to draw attention to the fact that it occurs in West Africa. REFERENCE Casretant, A., and Cuarmers, A. J. (1919). Manual of Tropical Medicine, Third Edition, pp- 1747-8. London: Bailliere, Tindall and Cox. 283 A FUNGUS OF THE GENUS WOCARDIA CULTIVATED FROM HEART BLOOD BY J. W. S. MACFIE AND A. INGRAM (Recezved for publication 24 August, 1921) The fungus here briefly described was cultivated from blood withdrawn from the heart at the autopsy on a patient who died in May, 1920, at Accra, of an obscure complaint. The notes of the case are as follows :— The deceased was a native man, about twenty-five years of age, regarded clinically as possibly suffering from encephalitis lethargica, and said to have been ill six days before death. He was stated to have shown paretic symptoms of the arms, and to have had convulsive seizures during which there was opisthotonus. Consciousness was lost temporarily. during the illness, but was recovered before death. At the autopsy, which was made on the 14th of May, 1920, eight hours after death, the body was found to be well nourished ; post- mortem rigidity was present in the legs and arms but absent from the neck. There was an old scar over the external aspect of the right upper arm immediately above the elbow joint, an old circular cicatrix about the size of a sixpenny piece over the left malar bone, and a small punctured wound at the back of the neck on a line with the thyroid cartilage: the latter wound was oozing blood, and may have been inflicted after death when placing the body in the coffin. On examining the internal organs, both lungs were found to be affected with broncho-pneumonia: the left lung was adherent to the 284 parietal wall by recent fibrinous formations. Brain: meninges somewhat congested; blood vessels in sulci engorged; no pus on the surface nor at the base. Heart: blood fluid; no abnormality noted. Cultures were made with blood obtained from the heart by puncturing the organ after searing its surface and taking every precaution to avoid contamination: in these cultures the Nocardia was grown. Digestive track, spleen, kidney, and liver: congested, but otherwise showed no apparent pathological condition. The organism cultivated from the heart blood of this case grew well on blood agar and ‘nasgar’ at 37°C. and at the temperature of the laboratory, about 26°C., producing a somewhat slowly spreading growth which was very firmly adherent to the medium. The colonies were at first smooth and dome-shaped, but later became puckered and opaque in the middle and radially striated and semi-transparent at the periphery. After a few days an efflorescence appeared on the older parts of the growth which in the earliest cultures was abundant and blue, and in later sub- cultures was scanty and grey or white. The medium sublying the growth was not stained. The organism grew both aerobically and anaerobically, but slowly and rather feebly under the latter conditions. The cultures had no distinctive odour. When examined microscopically the growth was seen to be composed of freely branching non-septate hyphae about 1p or less in diameter. In old cultures many of the hyphae were more or less fragmented, and the ends of some of the filaments were slightly thickened and club-shaped. The organism was Gram-positive but not acid fast. It grew well on blood-agar and ‘nasgar,’ as already stated. When first isolated it grew scantily on agar, but subcultures were not successful. It did not grow on glucose agar or maltose agar. On potato it produced an abundant whiteish growth which developed a brown or grey-brown efflorescence and stained the medium dark brown. On blood serum it grew well, causing no liquefaction but staining the medium dark brown. It did not grow on gelatin, nor in ordinary broth and peptone water. Its qualitative bio-chemical reactions were tested, and no change was produced in any of the following:—Arabinose, Rhamnose (iso-dulcite), Galactose, Glucose, Laevulose, Mannose, Lactose, 285 Maltose, Saccharose, Raffinose, Amylum, Dextrin, Glycogen, Inulin, Amygdalin, Helicin, Phlorrhizin, Salicin, Glycerol, Erythrol, Adonitol, Dulcitol, Inosite, Mannitol, Sorbitol, and Litmus milk. A guinea-pig inoculated intraperitoneally with an emulsion of a culture appeared to be unaffected. This experiment, however, was not made until seven months after the organism was isolated. The organism showed the characters of a fungus of the Genus Nocardia, but so far as we are able to ascertain, does not correspond with any of the numerous species already described. Although it was obtained in cultures made from blood aspirated from the heart, it is not possible to say if the organism is pathogenic to man, but the fact that its growth was almost restricted to potato and media containing either blood serum or ascitic fluid is perhaps significant. We propose for this fungus the name Nocardia cruoris. SUPPLEMENTARY NOTE ON A CASE OF BRONCHOMONILIASIS IN A NATIVE OF THE GOLD COAST A short time ago (1921) we described a fungus of the Genus Monilia which we had isolated from a case of bronchomoniliasis at Accra. The fungus belonged to the Tvopicalis group of Castellani and Chalmers, and closely resembled in its bio-chemical reactions M. nivea, but we could not say at that time whether it was actually the same or distinct, because we were unable to test its action on Raffinose. We are now able to fill in this gap in our description, and to state that neither acid nor gas is produced in Raffinose. The fungus, at the time this reaction was tested, had been isolated and maintained in cultures in the laboratory for about eight months, but a re-examination of its other bio-chemical reactions showed that they had not appreciably altered. The organism, therefore, cannot be regarded as M. nivea, and as previously pointed out, it differs from all the other members of the TZvoficalis group; it must, therefore, be regarded as a new species, for which we propose the name Monilia accraensis. 286 In support of the view that the failure of the fungus to ferment Raffinose was not due to the loss of this property after isolation from the human host, it may be added that we have recently obtained the same organism from the sputum of another native patient at Accra, and that when tested immediately after isolation it produced neither acid nor gas in this medium. REFERENCE Macrir, J. W. S., and Incram, A. (1921). Bronchomoniliasis Complicating Pulmonary Tuberculosis in a native of the Gold Coast, West Africa. Annals of Trop. Med. & Parasitol.. XV, pp. 53-58. 287 REPORT ON RAT-FLEA INVESTIGATION’ BY R. NEWSTEAD, F.R.S. AND ALWEN M. EVANS, M.Sc. ‘Received for publication 25 August, 1921) INTRODUCTION The investigation was carried out in conjunction with Dr. E. W. Hope, Medical Officer of Health for the City of Liverpool, and his Assistant, Dr. W. Hanna, whose material assistance has made it possible for us to carry it to a successful conclusion. The object of the investigation was to determine the distribution of the various species of fleas occurring on the rats in the Port and City of Liverpool, with special reference to those species which are responsible for the transmission of plague from rat to rat and from rat to man. The period during which the investigation was carried out extended from April 12th, 1920, to April 12th, 1921, rats being examined daily five times each week, except during the month of August and from December 22nd, 1920, to January 12th, 1921. TECHNIQUE As a preliminary to these investigations, field observations were conducted as to the general methods adopted by the professional rat catchers in the capture of the animals on board ship and in the city. It was then decided that the rats sent to the laboratory for examination were to be taken alive from the traps, and placed singly in strong calico bags. The bags were subsequently placed in a lethal chamber, where they remained until the rats and the fleas upon them were dead. This method ensured that all the fleas living on each rat at the time of capture were secured. It was further * Reprinted from The Annual Report of the Medical Officer of Health (Liverpool) to the Port Sanitary Authority, for the year 1920, by permission of Prof. E. W. Hope, M.D., with additional charts, etc. 288 decided that, as far as possible, only rats caught singly in traps were to be sent to the laboratory, because it was seen that, when a> number of rats are together in one trap, sweating is apt to occur, with the result that the fleas leave the rats before the latter are captured. LABORATORY METHODS The fleas were collected from each bag and rat, over a sheet of white paper, and placed in a numbered watch-glass containing glycerine. In this medium they were arranged in rows on a slide, and identified by microscopical examination. DEFINITION OF ZONES ZONE I.—SHIPs. The number of ships from which rats were received was one hundred and twenty-five. The ports from which the ships sailed are named in Table I (wede infra). ZONE II.—Docks, ETC. This Zone comprises that part of the Port of Liverpool lying west of a certain line, which defined as accurately as possible the eastward boundary of the warehouse area of the Port. (a) Docks. This section of Zone II is limited to the wharves themselves and the sheds actually situated on the Docks. (6) Warehouses. This section includes the rest of Zone II; the majority of the buildings, from which rats were received, were warehouses. ZONE II].—City. All of the City east of the line bounding Zone II is included in this Zone. THE SPECIES OF RATS No attempt is made to separate the varieties of the species Mus rattus and Mus norvegicus. Mus rattus and its varieties are included under the heading ‘black rat,’ and Mus norvegicus and any varieties are referred to as ‘ brown rats.’ As will be seen from the following table, all the rats received from ships were black. In Zone IA the black rats outnumbered the brown Showing the number of rats received from ships sailing from the various ports. 289 Taste I. Region Mediterranean , India, Ceylon, E. Indies, Burma East Africa... West Coast of Africa West Coast of S. America Brazil Argentine ... North America * Within the Tropics. Port No. of rats -| Alexandria %C 64 Constantinople ... 5 Costanza 2 Genoa 5 Salonika ... 4 hfe} -| Bombay*... 5 Calcutta* 18 Colombo* 4 Rangoon* 47 Java* I 75 -| Beira* I Mombasa* 3 Various Ports I 5 -| Various Ports 59 -| Valparaiso oa 5 Callao .| 13 Coronel ... 5 18 Pisco* 3 Talcahuano 3 Various Ports 33 75 -| Bahia* I Pernambuco* 18 Rio de Janeiro*... . 3 Rio Grande 5 37 -| Buenos Aires 53 R. Plate ... 17 Rosario 15 85 -| Galveston 14 New Orleans 40 New York I 55 290 rats by nearly five to one. In Zone IIB, the black rats were much more numerous than the brown rats. In Zone III, however, the position is more than reversed, the brown rats being nearly nine times as numerous as the black rats. Taste II. Showing the distribution of black and brown rats. Zone Black.rats Brown rats Total I 50 a5 a cee ote 469 — 469 PIA, <:.- nee 23 a5 one 24 5 29 Tip) =.-- 355 ano oes os 442 274 716 Lit Bae aor Sac aa ay 20 179 199 Torals ... 2A: 955 458 1,413 Tance IIT. Showing distribution of the various species of fleas. Zone Xenopsylla | Ceratophyllus| Ceratopbyllus| Leptopsylla | Ctenocephalus| Totals cheopis fasciatus londiniensis muscult Canis 1B tas oo 489 219 — 8 — 706 IIa fe) 75 _ 9 _ an IIs 60* 1,510 3 326 = 1,899 Tl acc ane 3 Pi eos A 12 i Io I 346 Torats AW 2,124 15 353 I 3,045 * This figure includes 56 specimens from No. 32 T- Street. OBSERVATIONS ON THE VARIOUS SPECIES OF FLEAS FOUND Xenopsylla cheopis, Rothsch. The Indian Plague flea is ‘the common rat flea of the tropics’ (Rothschild, 1910). It is prevalent also in sub-tropical countries, and ‘is common during summer and autumn in some of the warmer parts of the temperate zone, more especially in ports which have maritime intercourse with the tropics’ (Chick and Martin, 1911). It has been demonstrated experimentally by Bacot (1914) that temperatures below 40° F. are fatal to all but the imaginal stages 291 of this species. Its almost complete absence from more northern latitudes is therefore attributed to the low winter temperatures which prevail there. In England, X. cheopis has been recorded from a brown rat at Plymouth (one specimen) (Rothschild, 1905); from a plague-infected brown rat at Bristol (two specimens), 1916, taken in a rag factory that was the seat of an outbreak of plague; and in 1911 from Guy’s Hospital, London, where the species had established itself on a colony of brown rats. In the last case, it was found that the infested rats were living in an artificially heated environ- ment, beneath the laboratory, and the presence of this, the first flourishing colony of X. cheofis to be found in England, was attributed to this fact. SEASONAL VARIATION (Chart II and Table VII). Zone I is the only Zone from which a sufficient number of specimens of X. cheopis have been found to allow of any deductions as to seasonal variation. The curve, Chart II, has no very well marked characteristics; the highest point reached, June, 1920, rises from low figures in both May and July. The months September, October, November, show a second but not so great elevation, falling to the lowest figure in December. In view of the prophetic statement put forward by Chick and Martin (1911, p. 125}, it is interesting to observe that we have found a number of X. cheofizs on the ship rats during every month in the year, the lowest being 0°30 in December, 1921, and the highest 2°18 in June—not in September, as might have been expected. Owing to the large numbers of Xenopsylla cheopis which occur on ship rats, it would appear likely that this species might be present on some of the rats found in the dock sheds. This was found to be the case; out of the twenty-four rats received from Zone IIA, three were found to carry X. cheopis. The data were as follows :— Taste IV. Rat No Date nS Se Building X. cheopis Black | Brown 127 3I.V.20 I = S.H Dock Z 148 7-Ni.20 I _ S.C Dock 7 192 23-Vi-20 — \ I CG Dock I ToTar a 2 I 10 292 In Zone IIB and Zone III, apart from the colony of X. cheopis found in T—— Street, which is dealt with in a separate section, isolated specimens of this species were found in a few cases. The data are as follows :— Tape V. Zone ITz. Rat No. Date Building X. cheopis Black Brown 101g 21.121 _ I 14 C—— Street I 1127 10.11.21 _ I 29 C—— Street I 1135 IL.U.21 -- I 27 K—— Street I 1417 12.1V.21 — I M— Mills I | Totar fo) 4 4 Zone III. Rat No. Date | Building | X. cheopis Black Brown | | 1068 | 314.21 — I 5B Road | I | | 1gor* | S.iv.21 I — W— Village 2 i) . Tora I 1 | 3 Ceratophyllus fasciatus, Bosc. This is the common rat flea of temperate countries. In the rural districts of Suffolk and North Essex, Strickland and Merriman (1913) found this species to comprise 60 per cent. of the flea population of rats. Our figures show a much higher percentage of C. fasciatus for the city of Liverpool :—78 per cent. in Zone IIB and 92 per cent. in Zone III. » SEASONAL VARIATION (Chart III and Table VII). The curve of frequency for Zone IIB reaches a markedly high level during the summer months, with elevations in May-June and again in as September. A distinct depression occurs in July. The temperature curve (Chart I), on the other hand, is considerably higher in this latter month than in May. In September it is lower than in July. The frequency curve for Zone III is based on a much smaller number of records than that for Zone IIB, but with one exception it possesses the same characteristics as the latter. The exception occurs in January, when it rises to 4:11. This high figure is due to the occurrence of two very heavily infested rats among a total of nine. The average infestation of the other seven rats was only 0°7. It is apparent that there are two periods of maximum prevalence of the species, one in early summer, and a second in September. (The latter may possibly begin during August.) These periods both occur during the warmer half of the year, but the lack of detailed correlation between the frequency and temperature curves makes it clear that in Liverpool the prevailing atmospheric temperature has not such a direct influence on the prevalence of the species, as results obtained elsewhere in this country have suggested. There is no. correlation between the curves of average humidity and frequency of C. fasciatus. It must be noted, however, that the former is based on records taken in the open, and that the atmospheric humidity probably differs considerably in the buildings, sewers, etc., frequented by rats. Hosts. All the rats from Zone I were black, therefore no comparison between the number of fleas found on the different species of rats can be made for this Zone. In Zone II the infestation of brown rats with C. fasciatus was rather heavier than that of the _ black rats. Leplopsylla musculi, Dugés This is a widely distributed species naturally parasitic on mice (Mus musculus), but frequently found on rats where the latter inhabit buildings, etc., frequented by mice. This species is a proved potential carrier of plague bacillus, and ‘a small percentage’ will, according to Bacot (191y), ‘bite man under certain conditions.’ Strickland and Martin (1913) found it to comprise a very small percentage of the flea fauna of rats in East Suffolk and North Essex, only three specimens being taken in all. Bacot (1919), however, considers that this species may be more prevalent in England than 294 this figure suggests. As the following figures show, this is the case in the city of Liverpool :— } per cent. L. muscult. Zone IIA Hee ate ie 0°7 per cent. of total. Zone IIB nce she As 172 oe, % Zone III = ae ee 2°8 ” ” SEASONAL VARIATION (Chart ITV and Table VID). The curve shows that the species is most prevalent in Zone II during the months of June-September, after which the numbers fall away to almost zero during December-April. This curve agrees largely with that for C. fasciatus in Zone IIB, but the rise takes place a month later, and the disappearance during the winter months is more complete. As in the case of C. fasciatus, the curve rises to a high level during September in spite of the drop in temperature during this month. Hosts. The numbers of L. musculi per rat were almost equal on the two species, the averages were :— Zone I].—Brown rat ... Number of fleas perrat 0°41 Blackrates on: - Be sts 0°48 We —Brownbateses ey = a 0705 Black rat+« ... fe 5 a 0705 Ceratophyllus londiniensis, Roths. This species is said to be ‘A rare Mediterranean species, probably introduced by port rats, has occurred in the house mouse (Mus musculus), and possibly on the brown rat (E pimys norvegicus) in London, Dover and Aberdeen.’ (Rothschild, 1911.) Its distribution in Liverpool is interesting; apart from two isolated cases in Zone II, the records, four in all, came from a certain area, where the species appears to have established itself. Ctenocephalus canis, Curtis The common dog flea. This species has frequently been recorded from rats, sometimes in large numbers. It was therefore surprising to find only one specimen, which occurred on a brown ‘rat from Zone III, 295 Ctenopthalmus agyrtes, Heller Strickland and Merriman, and Nuttall and Strickland, working in rural districts of East Anglia, found this species to comprise a large proportion of the rat fleas in this part of England. Our records, however, show a complete absence of this species. Rothschild (1915) states that this species occurs on the ‘ brown rat jiving in the fields, and on the bank vole .'. . the common ° shrew . . . and others.’ It is not, therefore, surprising that it is not found on rats in such an urban district as the City of Liverpool. Further, ten other species of Pulicidae, which are recorded by the above-named authors, from rats in East Anglia, and which are naturally parasitic on birds and small wild mammals, are likewise absent from our records. PERMANENT BREEDING PLACES OF XENOPSYLLA CHEOPIS On the last day of January, 1921, a specimen of X. cheopis was found on a brown rat from No. 32, T Street. We requested that more rats should be sent from this building, and altogether we received during the first half of February thirteen drow rats from which no fewer than fifty-six X. cheopis were taken. The records were as follows :— ‘Tasre VI. | No. Date Brown : X. cheopis C. fasciatus 1067 314.21 I I _ 1087 3.1.21 I 5 2 1094 441.21 I 5 7 1095 441.21 I 5 4 1096 4.1.21 I 9 4 1097 4.11.21 I 7 2 1103 7-li.21 I 6 ° 1104 7-ii.21 I 2 8 1105 7-ii.21 I 6 4 1106 7.21 I I ° 1107 7-li.21 I 3 4 1108 7-11.21 I ) 5 2 1133 11.20 I I _ Torar 13 56 37 Average X. cheopis per rat = 4°3. 296 There can be no doubt that here at T——\ Street is a second ‘ flourishing colony of X. cheopis in the British Islands.’ As in the case of the first such colony to be discovered (Rothschild, 1911), its existence 1s probably to be attributed to the fact that its hosts were living in an artificially heated environment. Beneath the middle of the roadway adjoining the premises T—— Street runs a steam culvert. The excavations, which it was possible to make, revealed that the rat burrows lead in the direction of the culvert. It was, therefore, thought probable, that the nests Tasre VIL. Showing the average frequency during each month of the common species in the zones in which they occur. F Zonr I Zone II (b) Zone II Average Mean Average Date Tempera- | Humidity Xexuiop- Cerato- Cerato- Leptop- Cerato- ture sylla phyllus phyllus sylla phyllus cheopis | fasciatus | fasciatus | muscult | fasciatus 1920— April or 46°3° 73 0°33 1°75 12°66* oro No rats received May 4) 52309 74 0°59 o"90 5°38 0°32 ny June Foren Asa Th 2°18 o*40 3°28 1°84 3°27 July ee| PS see 79 o°51 o16 2°70 o89 l'o7 August ...) 564° | 80 Investigatijon suspend/ed. September Age 81 1°58 folze) S17 1°83 BEY October es ersaers 84 rz5 0°22 1°82 0°46 10g November...|_ 45°8° 83 118 0°37 161 o'4l 0°68 December 40°5° 86 0°30 0°09 129 0°06 o58 1921— | January ...| 46°0° 84 | 048 0°03 119 0°04. 4qu* | | February -..) 401° | 84 0°93 O75 1°33 o"o4 ry Marchiiiieccs|| so 45c5- 79 0°90 0'70 108 0°07 1°00 April pe-| | 46°77” 74 1°00 _— o61 0°09 o'40 | * Based on a very small number of records. 257, 4 of the rats were situated in the near neighbourhood of the steam culvert, and as the temperature of the latter was found on 21.2.21 to be 105° F., the area surrounding it for a considerable distance must be maintained at a temperature higher than the normal. Unfortunately it was impossible to trace the burrows far enough to discern whether the nests were actually within the area influenced by the steam culvert, and, therefore, the suggested explanation of the presence of this colony of X. cheopzs can only be regarded as an extremely probable one. SUMMARY 1. Five species of fleas were found to occur on rats from the ships, Port, and City of Liverpool. They were :—Xenopsylla cheopis, Ceratophyllus fasciatus, Leptopsylla musculi, Ceratophyllus londiniensis, and Ctenocephalus canis. 2. Xenopsylla cheopis occurred freely on ship rats throughout the whole period of the investigation. It was also found on three rats from the dock sheds, and isolated specimens were found on four rats from Zone IIB, and on two rats from Zone III]. A permanent breeding place of the species was discovered in certain premises in Zone IIb. 3. Ceratophyllus fasciatus was universally prevalent during the whole course of the investigation. The number of fleas per rat was greatest during the summer months, but the curve of frequency could not be correlated in detail with that of the average temperature. 4. Leptopsylla musculi was most prevalent on rats from Zone II. It occurred very rarely upon ship rats. Ceratophyllus londiniensis was found rarely in Zones II and III, and of Ctenocephalus canis one specimen was taken in Zone III. 60'0 April May June July Aug. Sept. Oct. Nov. Dec. Jan. Feb. Mar. April Black line :—Average relative humidity. Dotted line :—Average temperature. Cuarr Il. Xenopsyila cheopis 0-0 April May June July Aug. Sept. Oct. Nov. Dec. Jan. Feb. Mar. April Average number of fleas per rat in Zone I. 299 Cuart Ill. Cerarophyilus fasciatus 5-0 4:0 3:0 2:0 1:0 0:0 April May June July Aug. Sept. Oct. Nov. Dec. Jan. Feb. Mar. April Black line :—Average number of fleas per rat in Zone IIs. Dotted line :—Average number of fleas per rat in Zone III. 300 Cuart IV. Leptopsylla musculi April May June July Aug. Sept. Oct. Nov. Dec. Jan. Feb. Mar. April Average number of fleas per rat in Zone IIs. REFERENCES Bacot, A. W. (1919). Fleas and Rats in relation to Plague. “fourn. Roy. Sanit. Inst., Vol. XL, No. 1, 1919, pp. 53-60. ——-—- (1914). A study of the bionomics of the common rat fleas and other species associated with human habitation, with special reference to the influence of temperature and humidity at various periods of the life history of the insect. Fourn. Hygiene, Plague Supplt. IIT, Jan. 14, 1914. Cuicx, H., and Marrin, C. J. (1911). The fleas common on rats in different parts of the world, and the readiness with which they bite man. Fourn. Hygiene, Vol. XI, 1911, pp- 122-136. Martin, C. J., and Rowranp (1914). Rat Plague in East Suffolk. Reports to Local Govern- ment Board on Public Health and medical subjects. Nurtatt, G. H., Srrrcxranp, C., and Merriman, G. (1913). Observations on Pritish Rat- fleas. Parasitolozy, Vol. V1, No. 1, April 17, 1913, pp. 1-19. Rotuscuirp, N. C. (1905). Occurrence of Pulex cheopis Rothsch, at Plymouth. Ent. Mo. Mazg., Ser. 2, Vol. XVI, p. 139. (1910). A synopsis of the fleas found on Mus norvegicus decumanus, Mus rattus alexanérinus, and Mus musculus, Bull. Ent., Res. 1, pp. 89-98. — (1911). A further note on Xenopsylla cheopis, Rothsch. Ent. Mo, Mag., Ser. 2, Vol. XXIL, p. 113. (1915). A synopsis of the British Siphonaptera. Ent. Mo. Mag., Ser. 3, Vol. I, pp- 49-112, pls. VII, XIV. ——— (1916). The occurrence of Xenopsylla cheopis, Roths., in Bristol. Ent. Mo. Mag., Ser. 3, Vol. II, p. 279. 301 NATURAL ENEMIES OF STEGOMYIA CALOPUS, Mezicen BY CF ive YOUNG From the Laboratory of the Liverpool School of Tropical Medicine, Mandos (Received for publication 25 July, 1921) PLATES XIX AND XX During the examination of various collections of water in Manaos for the larvae of Stegomyia calopus, their absence from many apparently suitable breeding-places attracted attention. Larvae were found capable of developing in water taken from some of these places, except when certain insects were present. A collection of various aquatic insects from ponds, igarapés, etc., was therefore made and placed in jars of water together with the larvae of S. calopus. Under these conditions the following were found to destroy Stegomyza larvae with varying degrees of efficiency: larvae of dragonflies, water bugs (Hemzftera) and their larvae, larvae of water beetles, and two others not identified. Of these the larvae of dragonflies and water bugs were found to be the most destructive to the larvae of S. calopfus in captivity. The habits of S. calopfus larvae, which constantly range about in search of food, render them particularly liable to attack by predaceous insects, as compared to certain Culex and Anopheles larvae which may remain stationary at the surface for long periods. The commonest mosquito larvae found in Mandaos were those of Siegomyia calopus and Culex fatigans; these two species were frequently discovered breeding together in wells, barrels and other collections of water. The movements of both larvae and pupae of C. fatigans differ from those of S. calopus in being much quicker, thus rendering them less easily caught by predaceous insects. The larvae of C. fatigans spend most of their time at the surface, but 302 descend occasionally. On the other hand, the larvae of S. calopus, although occasionally searching the surface film for food, while taking in air, spend most of their time ranging over the bottom and sides of their habitat and are thus particularly exposed to attack by the dragonfly larva, which hes in wait at various depths below the surface. DRAGONFLY LARVAE Dragonflies are present in Manaos throughout the year, there being no cold season, and larvae were readily found at all times. Although representatives of the AGRIONIDAE have been found in Mandaos, members of the AESCHNIDAE and LIBELLULIDAE have been used exclusively in this work. Two species of each of these families are shown (Plate XX, figs. 4 to 7). The larvae of the LIBELLULIDAE (Plate XX, figs. 4 and 5) differ from those of the AESCHNIDAE (Plate XX, figs. 6 and 7) in having the abdomen shorter than the hind legs. According to Miall (1912), the larvae feed on insects, snails, tadpoles, and small fishes. In the laboratory they were observed to attack other aquatic insects, including members of their own species, water bugs and also tadpoles. I have never observed them attempt to seize any object which was not moving. Larvae have been observed resting on plants and on the sides and bottoms of .artificial ponds and fountains, igarapés and other natural collections of water. They usually remain motionless and await the approach of suitable prey, which they seize by suddenly shooting out the labium. This is carried on a jointed arm which lies below the head and thorax when not extended.’ The victim is then broken up rapidly by the jaws and swallowed, only three or four seconds being required for disposal of a Stegomyza larva. When the moving object is out of range of the labium they sometimes spring towards it by squirting water from the rectum. When disturbed they also use this method of moving to a greater depth. Respiration is carried out by drawing water into the rectum, but the more fully developed larvae are said to be able to take in air through the thoracic spiracles and, in the case of Aeschnid larvae, directly into the rectum (Miall, 1912). They are able to live in comparatively foul water. A dragonfly cog deposited eggs in a barrel containing thick greenish water with traces of oil on the surface and sides of the barrel and decomposing vegetable matter at the bottom. Larvae of the type shown on Plate XX, fig. 5, developed and lived for at least thirty-nine days, by which time they had reached a size of 2 cms. in length. Observa- tions could not be continued. No dragonfly larvae were found in water which did not receive rain. One of the dragonfiy larvae selected for experiment was identified as Pantala flavescens, F., by Dr. G. A. K. Marshall, who describes it as a ‘migratory and almost cosmopolitan species.’ It was found in several of the artificial ponds and fountains in public squares in Manaos (Plate XIX, figs. 1 and 2), in the igarapés, and by Dr. R. M. Gordon in a broken drainpipe under the pavement of one of the public thoroughfares (Plate XX, fig. 3). In the pond shown (Plate XIX, fig. 1) they were constantly present along with various other species during the period of ten months that they were observed. They destroyed S. calopus larvae and pupae of all sizes, usually disposing of the larger ones first. Six undetermined species of dragonfly larvae were also experimented with, all of which proved destructive to the larvae of S. calopus. WATER BUGS, HEMIPTERA The distribution of the water bugs (Zaitha spp.) used in the experiments is somewhat similar to that of the dragonfly larvae, the former having been found occasionally in the pond shown in Plate X1X, fig, 1, and in the igarapés. They are not, however, so plentiful, and difficulty was experienced in obtaining a supply at times. They fed on aquatic and other insects, such as grasshoppers, etc., when placed on the surface of the water, but only when living and showing movement. They also attack their own species. They suck the body juices of their victims, a specimen measuring 1°3 cms. in length taking anything from five to fifteen minutes to dispose of a fully grown S. calopus larva. The water bugs are thus consider- ably slower than the dragonfly larvae, but can continue feeding for a longer period. They usually rest on plants or sides of ponds with the posterior end of the body at the surface, the head downwards 304 and the front pair of legs hanging free. When a suitable victim approaches it springs on it by a quick movement of the hind legs and seizes it with the fore legs which are adapted for clasping. When the captured insect is large all the legs may be used for gripping, and at least three mosquito larvae can be held at a time. Both adult and larval water bugs fed freely on Culicine larvae. FEEDING EXPERIMENTS Experiments were carried out to test the relative efficiency of dragonfly larvae and water bugs under artificial conditions. Larvae of P. flavescens, measuring from 1°4 to 271 cms., and water bug larvae of 1°1 to 1°9 cms. in length, were used. Large cylindrical glass jars containing two litres of tapwater were employed, the depth of water in the jar being 10 cms. Twenty fully grown mosquito larvae were used in each experiment and were introduced before the insects to be tested, of which two were always employed. The temperature of the water varied between 28°5° and 32°C. The jar was observed continuously for the first half hour, and at three-quarter, one, one and a quarter, one and a half, two, two and a half, three and five hours, and the number of larvae destroyed during each period recorded. A control jar was used containing twenty mosquito larvae similar to those used in the experimental jar. Control larvae died in one experiment only, and this is not included below. In the graphs shown in text-figs. I and 2 the ordinals represent larvae destroyed and the abscissae time in hours. The curves in text-fig. 1 represent the averages of two Fic. I. 16 12 Hours 1 2 3 1 5 Larvae seseeeeeee = 2 dragon fly larvae. —-=2 water bug larvae. ----=1dragon fly + 1 water bug larva. S. = 20 larvae of S. calopus. C. = 20 larvae of C. fatigans. 395 experiments in each case, but in fig. 2 each curve is constructed from one experiment. It will be observed in text-fig. 1 that fastest time was made by dragonfly larvae with S. calopus and second fastest by a combination of dragonfly, water bug and Stegomyza larvae. The other curves show only slight differences. In fig. 2 a mixture of Fic. 2. 8.+C. 8.+C. §.+C. ie] Larvae w Hours 1 2 3 saacseaee = 2 dragon fiy larvae. = 2 water bug larvae. ---—-=1 dragon fly + 1 water bug larva. =, pared ay ey equal numbers of S. calopus and C. fatigans larvae were used, and here a combination of dragonfly with water bug larvae was the most effective. In the dotted curve the two dragonfly larvae destroyed all the S. calopus in three minutes, but took five hours to the remaining Culex larvae. So far as rate of destruction was concerned, therefore, under these artificial conditions, dragonfly larvae alone were the most effective with S. calofus larvae, and a combination of water bug and dragonfly larvae with those of C. fatigans. As regards quantity, the largest number of fully grown S. calopus larvae destroyed by a larva of P. flavescens in twenty-four hours was one hundred and fifty-six. This larva during five days, being fed on the first, third and fifth days only, consumed three hundred and seventy-nine larvae. The largest number destroyed by a water bug was one hundred and six in twenty-four hours, but only half of these were killed for feeding purposes; a commoner figure is about sixty. When about to moult these insects cease feeding, so that the numbers consumed in twenty-four hours varied from o to the maxima mentioned. 306 Some experiments with dragonfly larvae were carried out under natural conditions. An uncovered cement tank, measuring I metre square by 78 cms. deep, in a backyard was filled with water to within 10 cms. of the top. Between one thousand one hundred and one thousand two hundred mosquito larvae in all stages of development were introduced. The great majority of these were S. calopus, but a number of C. fat2gans were also included. Pupae and eggs were also present. As a control, a glass jar was filled with water from the tank and a few of both species of larvae added. At zero (five hours later) both species of larvae could be detected by a few moments inspection of the surface of the tank. Seven larvae of P. flavescens, varying in length from 2°3 to 1°3 cms., were then introduced. At sixteen hours both species of larvae could be detected. At twenty-four hours no S¢egomyza larvae could be found after fifteen minutes search, but Culex larvae were still present. At forty hours a few newly hatched Stegomyia and three Culex larvae cnly were found. One dragonfly larva was found dead. At forty-eight hours no larvae of any kind were found. At fifty hours the tank was emptied by syphoning out the water through several layers of gauze. No mosquito larvae were found. In the control jar, which had been kept beside the tank throughout the experiment, the larvae remained alive and active. The temperature of the water in the tank varied between 29'5° and 3315 Ge In the following experiment two barrels kept in an enclosure behind the laboratory were used. Although frequently receiving rain the water in these barrels was somewhat foul, particularly in one which had recently contained oil. In size they measured 85 cms. by 56 cms. at the widest part and the depth of water in each was 65 cms. Sétegomyia calopus was breeding naturally in both, large numbers of pupae and larvae in all stages of development being present. At zero (1 p.m.) five larve of P. flavescens were introduced into one barrel. At eighteen hours only small larvae were present. At twenty-four hours no larvae were found. 307 During this period larvae and pupae continued to be present in the control barrel to which the five dragonfly larvae were then transferred. ? At three hours pupae and larvae were still present. At seventeen hours only small larvae were present. At twenty-four hours a few small larvae were present. At forty-two hours no larvae were found. Thus two barrels were cleared of S¢egomyia larvae in twenty-four hours in one case, and in forty-two hours in the other, after the _ introduction of five dragonfly larvae. The latter were allowed to remain in the last-mentioned barrel, which contained the larvae of various other insects. During the succeeding twenty-six days, five dragonflies hatched, no Stegomyia larvae were observed, but on three occasions newly hatched Culex larvae, which did not reach the pupal stage, were found. During the last nineteen days of this period Stegomyia and Culex larvae were present in the other barrel. No Stegomyia \arvae were found in Mandos in any water containing dragonfly larvae. A certain artificial well which contained dragonfly larvae and was free from mosquito larvae, after being cleaned out was found to be breeding Culex fatigans and Stegomyia calopus, but no dragonfly larvae were discovered. Five weeks later Stegomyia larvae were absent and dragonfly larvae were again found. A few Culex jatigans larvae were also present. THE DRAGONFLY AS AN ENEMY OF THE ADULT MOSQUITO It was thought probable that the adult dragonfly would prey upon Stegomyia calopus if. opportunity offered. In fine weather dragonflies are to be seen in the open places and larger streets of Manaos, and particularly around their breeding-places, as well as in the forest. During dull weather they are not so active, resting most of the time on the tops of shrubs and other objects, and rising only in pursuit of insects appearing close at hand. Both the dragonfly and Stegomyia calopus are, therefore, active in Manaos in bright warm weather, but the mosquito is also active at 308 night. Except for a few attracted into houses by lights, dragonflies were not observed at night. An attempt was made to see if they pursued S. calopus when given the opportunity. It was found possible by moving slowly to approach closely to the resting dragonfly without disturbing it, and S. calopus adults were then released from a glass tube at a distance of about a metre from it. The dragonfly nearly always pursued and caught the mosquito, usually returning to its perch consuming it. It was rot always possible, owing to the rapidity of its flight, to observe whether the dragonfly actually caught the mosquito, but in most cases it was seen to occur. Five S. calopus were released one at a time near a resting dragonfly; four at least were caught, and probably all five. The dragonfly was then caught, and the contents of the alimentary canal teased up in saline and examined microscopically. The mosquitoes were found to be broken up into such small particles as to be unrecognisable, the only indication of Stegomyia being the large number of flat scales present. Five different species of dragonfly, varying in length from 3 cms. to 6 cms., were tried against S. calopus, and all pursued and caught the mosquito. The larger ones were also observed to prey upon bluebottles and houseflies. SUMMARY Dragonflies and their larvae have been found to be destructive to Stegomyia calopus and their larvae respectively. Several other aquatic insects, including water bugs, have also been found to be inimical to S, calopus larvae. My thanks are due to the Director of the Laboratory, Dr. Wolferstan Thomas, for assistance and advice, and to Miss A. M. Evans and Dr. G. A. K. Marshall, Director of the Imperial Bureau of Entomology, for identification of the insects mentioned. REFERENCE Mratt, L. C. (1912). The Natural History of Aguatic Insects, Macmillan & Co. . c ’ . ARC «| «feos paps q pet ue m “9% Abie dG (i> ei4 . ) i 4 on “shill = ol TAS pen 310 EXPLANATION OF PLATE XIx Fig. 1. Gardens in public square in Manaos. Habitat of larvae of Pantala flavescens, F., and other dragonflies and water bugs. Pips: Habitat of larvae of Pandala flavescens, F. Annals Trop. Med. & Parasitol., Vol. XV PLATE XId iiges ii aie J” - ro : MA MIAN. 1 Vic aA NS t= N EXPLANATION OF PLATE XX Broken drain in street, Manaos. Habitat of larvae of Pantala flavescens, F. Larva of Pantala flavescens. Actual length 1°8 cms. Libellulid nymph. Actual length 1°8 cms. Aeschnid larva. Actual length 271 cms. Aeschnid nymph. Actual length 4°7 cms. Annals Trop. Med. & Parastol., Vol. XV PLATE Bich 4 Fic. Fic. Fic. F cal Tenlsne feo Ca. Fad _ Fond 313 WEST AFRICAN CERATOPOGONINAE BY A. INGRAM AND Jo Wr St MACFIE (Received for publication 24 August, 1921) PLATE XXI This paper is a continuation of the series of papers on the Ceratopogonine Midges of the Gold Coast already published in the Aynals of Tropical Medicine and Parasitology, and contains descriptions of additional new species. To some extent it is also supplementary, since we are now able to fill in a number of gaps in previous descriptions ; for example, particulars are given of the early stages and of the natural habitat of several of the species of Culicoides and Dasyhelea, facts which are of interest, because they point the way to means of control. It differs from the former papers, however, in that a few species are described which were collected, not in the Gold Coast, but in other parts of British West Africa. Many of the species belong to genera which were not considered in the previous series; but the number of species referable to each genus is small and insufficient to warrant the inclusion of detailed generic descriptions. For the same reason, keys to the species found in West Africa are not at present necessary. We have found it difficult in some cases to appraise at their proper value the characters employed by Kieffer in separating his smaller genera, and, where we have found only slight divergencies to occur, we have been in doubt whether it would be advisable, as would appear to be strictly necessary, to erect new genera. We regret that in dealing with these species we have not had the advantage of collaboration with Mr. Carter, whose recent appointment as Malariologist in Ceylon has unfortunately made it impossible for us to look to him for the assistance he was ever so ready to give. Some apology must be made for the figures which compare so unfavourably with those in the previous papers. In the present 314 paper they are mere outlines traced with the aid of a camera lucida. In figuring the hypopygium of the male we have not often attempted to show it as a whole, but for the sake of clearness and in consideration of our technical shortcomings, have drawn the various organs more or less separated, showing them, unless otherwise indicated, as they appear when seen in a ventral view. Since our earlier notes on the bionomics of these insects were published (1920) we have located another abundant source of Ceratopogonine midges, namely, the water lettuce P7stia stratiotes. This weed, which is exceedingly abundant in rivers, swamps, pools and lagoons in West Africa, and frequently covers large expanses of water, has for many years been recognised as a troublesome mosquito nursery and one very difficult to deal with, not only on account of the association with it of Mansontoides africanus but also because it provides protection between its leaves to the larvae of several species of Anopheles (e.g., A. costalis, A. mauritianus, A. nil), The very small number of observations which we have made hitherto prove beyond doubt that it is also a fruitful source of midges, for we have reared from it Cudécoides austeni, C. distincti- pennis, Dasyhelea inconspicuosa, Prionognathus pseudomaculipennis, and the species described in the following pages under the names Kempia ochrosoma, Eukraiohelea africana, E. versicolor, Probezzia pistiae and P. stephensi. Large numbers of Kempia ochrosoma, Eukraiohelea africana and Probezzia pistiae were procured from this source, so that it’ appears that these species, at any rate, are peculiarly associated with the plant. The types and co-types of the new species described have been deposited in the museum of the Liverpool School of Tropical Medicine. Genus CULICOIDES, Latr. Culicoides austeni, Carter, Ingram and Macfie. Five larvae of this species were found together with numerous larvae of Ochlerotatus irritans in a sample of water from a crab-hole. Three of the larvae were reared to the adult stage, the other two were killed and preserved. The larval and pupal pelts of the 315 specimens reared through to the adult stage were recovered. The following descriptions of the pupa and larva are based on these materials. PupaA. Length about 27 mm. Respiratory trumpets short and straight, tapering slightly towards the apex, raised on long stalks; rather strongly chitinised, middle third covered with large squamose spines; length of the trumpet about o'15 mm., length of the stalk about o'(05 mm. There are no knob-like processes, but the main tracheal trunk gives off in its distal third a continuous row of about nine short blunt processes. Cephalo-thorax: anterior marginal tubercle double, highly chitinised, the inner portion large, conical, and bearing a long strong bristle, the outer portion small, unarmed ; anterior dorsal very large, highly chitinised, irregularly conical, bearing two stout bristles; dorso-lateral smaller, bearing a long and a short hair; ventro-lateral an irregularly shaped tubercle bearing a long and a short hair and an apparently unarmed socket; ventro- median represented by a moderately long and a very minute hair. External to the ventro-median tubercle and a little posterior to the ventro-lateral is a small, unarmed, nipple-like tubercle which projects prominently outwards. Dorsal tubercles small: anterior single, bearing a minute blunt spine; posterior and lateral each bearing a hair. Immediately in front of, and slightly external to, the the anterior tubercle is a small tubercle which is apparently unarmed ; immediately behind, and slightly internal to the posterior, is a small tubercle bearing a very minute spine. Postero-dorsal tubercle small, bearing a hair, and behind it two unarmed sockets, Abdomen of the usual form. Anal segment terminating in two sharply pointed, divergent processes which are very highly chitinised, especially at their tips. Tubercles on the abdominal segments small, strongly chitinised ; arrangement and armature as in C. accraensis. LARVA. Length about 5'7 mm., greatest breadth about o°3 mm. Head: \ength about 02 mm., greatest breadth about 0°14 mm. Eyes large, bilobed. Bristles small, apparently arranged as in C. accraensis. Mental plate with a large, pointed central tooth. Hypopharyngeal sclerite moderately chitinised, bearing on each side usually eleven pointed, finger-like processes which are nearly equal in-size, excepting the fifth from the inner margin, which is slightly larger than the others. Mandibles well chitinised, pointed, with a 316 well-developed central tooth. _Body: hairs minute, terminal hairs on the anal segment small; anal gills of the usual form. — | GoLp Coast: Accra, April, 1921, reared from mud from pools and puddles near the station for the Weshiang Line (Pl. XXI, fig. 2); June, 1921, larvae found in water from a crab-hole. Christiansborg, July, 1921, reared from plants of the water weed_ Pistia stratiotes. Oblogo, June, 1921, reared from banana fibre. Culicoides distinctipennis, Aust. Although numerous specimens of this species have been collected at Accra, it is only recently that we have reared it from the early stages. In the samples from which it was reared were also. the early stages of several other species of Culicoides, and we were unable to identify with certainty the larvae. The pupa was, however, obtained, and is briefly described here. Pupa. Description based on a single pelt from which a male had emerged. Length 1:7 mm. Respiratory trumpets, raised on moderately long stalks; length of the trumpet about o'2 mm., length of the stalk about 0°04 mm. Distal extremity somewhat darkened, middle portion covered with squamose spines, proximal two-thirds bearing three small knob-like processes. Main tracheal trunk terminating distally in a fan-like arrangement of seven short, blunt processes. Cefhalo-thorax dark, operculum rather sparsely clothed with large, dark, squamose spines. Anterior marginal tubercle small, bearing a rather short, stout spine; anterior dorsal bearing a short, stout spine and a minute spine; dorso-lateral, small, bearing a hair and a short spine; ventro-lateral, small, bearing two hairs, one of which is quite short and spine-like ; ventro-median represented by a moderately long and a very small hair. Dorsal tubercles: anterior double, the two halves separated, the one situated anterior and slightly external to the other, each bearing a short, stout, dark- coloured spine; posterior, small and flat, bearing a minute spine; lateral, small, bearing a hair. Posterior to the lateral of the dorsal tubercles is an unarmed socket, and on the dorsum are several ill-defined darkened patches resembling flat, unarmed tubercles. Postero-dorsal tubercle small, bearing a hair and an unarmed socket. Abdomen of the usual form. Anal segment terminating 317 in two sharply pointed, somewhat’ divergent, dark-tipped processes which are not closely covered by squamose spines. Dorsal, ventro- lateral, and ventral tubercles as in C. accraensis. GOLD COAST: Accra, February to April, 1921; numerous specimens reared from moist soil and mud taken from the margins of pools and puddles near to the railway station on the Weshiang Line (Plate XXI, fig. 2). Oblogo, March, 1921; reared from plants of Pistia stratiotes taken from the river Densu (Plate XXI, fig. 1). Culicoides ertodendroni, C., 1. and M. In a previous paper a description was given of the female of this species. We are now in a position to add certain points in regard to the male. MeasurEMENTs (average of two). Length of body* ©... Sin, CES Se eeteewees whoo (73) SI, Length of wing or Sa xn rin one — 7 1 Oomme Greatest breadth of wing ... usd aa se at =. Of; mms In general, the description of the female applies also to the male, but the following points may be especially mentioned. Eyes separated. Antenna: last three flagellum segments sub-equal, the fifteenth being, however, slightly the longest and the fourteenth slightly the shortest. Scutellum bearing two central and two lateral bristles, and a few (six) short hairs. Wing: pale spot covering the anterior cross-vein rather diffuse and spreading almost to the costa, pale spots at the apex of the wing and along its posterior border almost or entirely absent; decumbent hairs scanty, but more than a single row in the basal portion of the wing between the fourth and fifth veins. Hyrorycium (fig. 1). . Ninth segment: tergite, posterior margin ‘slightly nctched in the middle and ending on each side in a well- developed conical process; sternite, deeply excavated. Forceps: side-pieces rather long and narrow, covered with relatively short hairs; claspers with a broad, hairy, basal portion, constricting abruptly to meet the terminal portion, which is of the usual form. Harpes (fig. 1 ay: moderately chitinised, somewhat strap-like, ee — = =e == * In all cases taken from the anterior margin of the thorax to the tip.of the abdomen of specimens mounted in carbolic. 318 tapering only slightly, with a short piece at the distal end bent ventrally or ventro-laterally lke the end of a cleek. dAedoeagus (fig. 1 6): form rather unusual; stem very short, highly chitinised, in ventral view shaped like the letter T; limbs long, moderately highly chitinised; ventral wall chitinised for only a short distance from the apex of the arch, membranous portion not spiculated. Fic. 1. Culicoides eriodendroni, C. 1. & M., outlines of male hypopygium, ventral view. a—forceps and harpes; b—aedoeagus. GOLD Coast: Nsawam, August and October, 1920; reared from larvae obtained from rot-holes in the stump of a silk-cotton tree and of ancther tree. Culicoides grahami, Aust. Several specimens of this species were reared from material taken from the base of a banana stump. The pupal pelt of one female was recovered, and as it was in some respects peculiar and showed several characteristic features, a brief description of it is given. Pupa.~ Length 1°6 mm., moderately well chitinised. Respira- tory trumpets long and curved; length about o' 17mm. _ Stalk or pedicle moderately long. The trumpets are of almost uniform width throughout their length, and are irregularly ringed. The main tracheal trunk gives off during its course through the trumpet a few short lateral branches which terminate in small tubercles scarcely projecting above the general surface level, and ends distally in a fan-like arrangement of the usual form. Cephalo-thorax: 319 anterior marginal tubercle large, covered with squamose spines, bilobed, and formed of an inner rounded portion and an outer conical portion, the latter bearing at its apex a long, strong bristle; anterior dorsal, double, each part bearing a long, stout bristle; anterior dorso-lateral, an irregular tubercle bearing two hairs; ventro-lateral, an irregular tubercle bearing two delicate hairs, ventro-median, apparently absent. The operculum is sparsely covered with squamose spines, most strongly developed along its periphery ; near its posterior margin, in the middle line, is a small elevation or tubercle covered with rather coarse squamose spines, but otherwise unarmed. Dorsal tubercles small; anterior, double, bearing a delicate hair internally and a small spine externally ; posterior, almost obsolete, bearing a small spine; lateral, bearing a hair. Postero-dorsal tubercle almost obsolete, bearing a delicate hair. Aédomen: anal segment bearing two transverse rows of relatively large spicules, the one near the anterior margin and the other about the middle of the segment; these spicules are developed most highly on the dorsal and lateral aspects. A group of similar spicules is present dorsally at the roots of the terminal processes. The terminal processes are short and pointed at their tips, they diverge almost at right angles, and their ends are turned dorsally. Tubercles on the abdominal segments poorly developed, and shaped like large spines. Dorsal tubercles: antero-submarginal, the inner bearing a short. spine and the outer a longer hair; postero-marginal, only a single tubercle present, situated posterior to the outer antero-submarginal tubercle, bearing a short spine. Ventro-lateral tubercles small, but little larger than the dorsal and ventral: antero- submarginal bearing a short spine; postero-marginal, the middle one bearing a hair, the other two short spines. Ventral tubercles : the middle one bearing a hair, the outer and inner each a short spine. Each abdominal segment (excluding the last, which has been referred to already) bears a transverse row of relatively well- developed spicules near its anterior margin; these spicules are most highly developed and most numerous on the more posterior segments. GoLp Coast: .Nsawam, 24th July, 1920; reared from material taken from the base of a banana stump. 320 Culicoides neavei, Aust. Several specimens of this species were reared from soft mud taken from the edges of pools at Accra. The larvae, together with those of several other species of Culicoides, frequented the mud which was very soft and almost semi-fluid. They were of the usual form, but we are unable to give details of their structure because, although we reared one adult from a larva isolated in a small tube, we did not succeed in recovering the larval pelt, and were, therefore, unable to distinguish amongst the numerous larvae in the sample those belonging to this particular species. The pupae also frequented the mud: the description which follows is based on an examination of the pelts of two pupae which had been isolated singly, and from which adult insects were procured. Pupa. Length about 1'9 mm. Operculum densely covered with dark brown squamose spines. Kesfiratory trumpets similar to those of Culecoides inornatipennis, short and raised on relatively long stalks; length of the trumpet about o'19 mm., length of the stalk about 0°05 mm. The trumpet is infuscated at its distal end, and also, slightly; at its base; it bears three or four quite small knob-like processes, the most distal of which is situated rather far anteriorly. The main tracheal trunk terminates distally in a hand-like group of about six short blunt processes. Cephalo-thorax: anterior marginal tubercle small, dark coloured, conical, bearing a relatively long stout spine which is directed ventrally, anterior dorsal well developed, conical, bearing a short stout spine and a minute spine; dorso-lateral prominent, bearing a hair and a short spine; ventro- lateral a rounded hump, bearing a short and a moderately long hair; ventro-median small, bearing a moderately long and a short hair. External to the ventro-median tubercle, and a little posterior to the ventro-lateral, is a small nipple-like tubercle, apparently unarmed. Just in front of and internal to the base of the stalk of the trumpet is a small hair. Dorsal tubercles: anterior double, the two parts being separate and well developed but not large knobs, the one situated anterior to the other, each bearing a short, stout, dark-coloured spine; posterior, poorly developed, bearing a minute spine; lateral, feebly developed, bearing a hair. In front of the anterior tubercle, and a little external to it, is an inconspicuous unarmed tubercle; posterior to the lateral tubercle is a socket-like 321 mark, apparently unarmed, and there is another similar mark situated more posteriorly. Postero-dorsal tubercle small, bearing a hair and two apparently unarmed, socket-like marks. Adédomen: anal segment with sharply-pointed terminal processes infuscated at their tips. Dorsal tubercles of the normal form: antero-submarginal, the inner bearing a short spine, and the outer a hair; postero- marginal, five in number, the outermost bearing a hair, the next a short spine, the innermost a minute spine, and the other two apparently unarmed. Lateral tubercles larger than the others and each with two sharp points, between which are the hairs or spines: antero-submarginal, bearing a spine; postero-marginal, the middle one bearing a hair, the other two, spines. Traces are visible of a second, rudimentary, antero-submarginal tubercle in a more dorso- lateral position. Ventral tubercles of normal form: the middle one bearing a hair, the other two, spines. GoLp Coast: Accra, February, 1921; reared from soft mud taken from the edges of pools and puddles near the station for the Weshiang railway (Pl. XXI, fig. 2). From the same material were reared also Culicoides austeni, C. distinctipennis, C. similis, C. schultzei and Stilobezzia spirogyrae. Culicoides similis, C., I. and M. At the time when this species was described (1920), the early stages were not known. They have since been collected, and are here briefly described. Pupa. Length about 1.9 mm. Operculum densely covered with dark brown squamose spines. Kesfiratory trumpets short and raised on rather long stalks; length of the trumpet about 0°19 mm., length of the stalk about 0:03 mm.. The trumpet bears on its proximal half three or four small knob-like processes which are infuscated. The distal end of the trumpet is dark brown: in it the main tracheal trunk terminates in a fan-like group of short, blunt processes. Cefhalo-thorax: anterior marginal tubercle dark brown, rather small, conical, bearing a relatively long, stout, dark-coloured spine; anterior dorsal prominent, irregularly conical, bearing a stout spine and a minute spine; dorso-lateral small, bearing a hair and a minute spine; ventro-lateral a rounded hump, bearing a small and a minute hair; ventro-median represented by two hairs, 322 one minute. Dorsal tubercles: anterior usually double (in one specimen single on one side), the two parts contiguous or separated ~ but almost side by side, well developed but not large knobs, each bearing a short, stout, dark-coloured spine; posterior, poorly developed, bearing a minute spine; lateral, poorly developed, bearing a hair. In front of the anterior tubercle, and a little external to it, is an unarmed tubercle; posterior to the lateral tubercle is a socket-like mark, apparently unarmed, and there are usually two similar marks situated more posteriorly. Postero-dorsal tubercle small, bearing a small hair and one or two apparently unarmed, socket-like marks. Aédomen: anal segment with sharply-pointed terminal processes infuscated at their tips; in the middle line, dorsally and posteriorly, is a small elevation covered with dark squamose spines. Dorsal tubercles of the normal form usually, but on some segments the outer two postero-marginal tubercles tend towards the form of the lateral tubercles: antero-submarginal, the inner bearing a short spine, and the outer a hair; postero-marginal, five in number, the outermost bearing a hair, the next a spine, the innermost a minute spine, and the other two apparently unarmed. Lateral tubercles larger than the others, and each with two sharp points between which arises the hair or spine: antero-submarginal, bearing a spine; postero-marginal, the middle one bearing a hair, the other two, spines. Traces are visible .of a second, rudimentary, antero-submarginal tubercle in a more dorso-lateral position. Ventral tubercles of normal form: the middle one bearing a hair, the other two, spines. : Larva. Although we did not succeed in recovering the larval pelt of any individual specimen isolated and reared through from the larval stage, we secured from the materials collected at Accra a number of larvae which we believe to be those of Culicoides similis. The larvae were found, together with pupae of C, szmlzs, in the soft, semi-fluid mud in the specimen jar at a time when this species was the only one emerging from the sample. Moreover, in three of the larvae, apparently almost ready to pupate, the pupal structures, including the respiratory trumpets and many of the cephalo-thoracic and abdominal tubercles, were clearly visible through the cuticle, and appeared to be identical with those of the pupa of C. simzlzs. 323 It is of interest to note the situations in which certain of the pupal structures were seen in the larvae. The trumpets lay in the first body segment, their proximal ends situated dorsally and laterally at the posterior end of the segment, and their free ends situated ventrally on each side of the middle line a little posterior to the head. The dorsal tubercles of the cephalo-thorax were situated dorsally at the anterior end of the second body segment. The terminal processes of the anal segment were turned so that they projected anteriorly, one on each side, with their tips directed -dorsally and situated at the anterior margin of the segment. The following description of the larva is based on the examina- tion of the three larvae alluded to as being apparently ready to pupate. Length about 3°5 mm. when fully grown, greatest breadth about 002 mm. Head, length about 0713 mm., greatest breadth about 008 mm. Eyes small. Bristles mostly small; on the ventral surface one pair, admedian, a little posterior to the hypopharyngeal sclerite, two pairs, admedian, almost contiguous, anterior to the hypopharyngeal sclerite, and one pair ventro-lateral; on the lateral surface two pairs, the one anterior and the other central; on the dorsal surface one pair admedian, anterior, two pairs sub- central, slightly separated, dorso-sublateral, two pairs, almost contiguous, posterior, dorso-lateral. Palpi and antennae well developed. Labium broad, blunt, dark coloured, apparently without teeth. Posterior margin.of hypopharyngeal sclerite bearing on each side ‘seven or eight sharply pointed, graded, teeth, the middle ones being the longest; these teeth are not very highly chitinised. Mandibles simple, pointed, without teeth. Body: appearing almost hairless, but actually bearing a few very small hairs; terminal hairs on the anal segment very small; anal gills of the usual form, deeply cleft distally into two pointed processes. GOLD CoAsT: Accra, February to April, 1921; numerous specimens of both sexes reared from soft mud taken from the edges of pools and puddles near the station for the Weshiang railway (Pl. XXI, fig. 2). Oblogo, February, 1921; a few specimens reared from sandy mud taken from the washing-place in the river Densu. 324 Culicotdes corsoni, sp.n. This insect, of which we possess at present only a single male, resembles in wing markings Culicoides similis and C. citroneus. From the former it differs in having no pale area covering the middle of the lower ramus of the fourth vein and-in the whole of the cross-vein being enveloped in a pale area; from the latter in having only a single pale area between the branches of the fifth vein, the small pale spot in the angle being absent. MEAsuUREMENTS, Length of body (one male) 588 Bhs as ee .. I'Omm, Length of wing... 398 ane bi he oo .. o8 mm. Greatest breadth of wing ... wl Eke te a .. 073 mm. Head : occiput dark brown. Eyes narrowly separated. Proboscis and palpi dark brown; the latter with the third segment somewhat inflated, especially in its basal two-thirds. Awtennae: infuscated; the thirteenth segment slightly longer than the terminal segment; the plumes only moderately developed. Thorax dark brown, with paler brown markings. Owing to the fact that the species was not recognised until the hypopygium had been examined in carbolic, exact details of the thoracic adornment cannot be given, but there appeared to be three large paler areas on each side of the anterior two-thirds of the dorsum, one in front and two behind. Scutellum yellowish-brown, somewhat darker at the sides; bearing one central and two lateral bristles, and a few (three) short hairs. Post- scutellum dark brown. Pleurae dark brown. Wexgs with pale markings resembling those of Cudicozdes citroneus. The pale spot in the middle of the anterior border covers the tip of the distal cell; there is no pale spot covering the middle of the lower ramus of the fourth vein, and the pale spots in the neighbourhood of the bifurcation of the fifth vein are arranged as in C. similes. Decumbent hairs scanty, not extending beyond the middle of the wing, and none being present in the anal angle, between the rami of the fifth vein, or at the base between the fourth and fifth veins. Halteres with creamy-yellow knobs. Legs infuscated, knees scarcely darker; tibiae with narrow pale bands basally and less distinct pale bands on the femora apically. A&domen dark brown. Hyrorycium. Ninth segment: tergite broad, bearing only a few stout hairs, the most notable of which are four arranged in a 325 transverse row near the posterior margin; posterior margin notched in the middle, and bearing at each lateral angle a long, slender, finger-like process about five times as long as it is broad at its base; apical lobe-like processes well developed. Sternite deeply excavated centrally. Forceps of the normal form; claspers highly chitinised, with rather blunt ends. Harpes resembling those of C. grahami, but with shorter distal portions. Aedoeagus Y-shaped, the limbs and the proximal end of the stem highly chitinised, the distal end of the stem more delicate and ending in a broad lip. The ventral wall of the aedoeagus is highly chitinised from the apex of the arch to about the middle of the limbs, but is not extended anteriorly and centrally as a spine; the membrane joining the aedoeagus to the ninth sternite is without spicules. The hypopygium resembles that of C. cztroneus, but the following differences, amongst others, may be noted. The long hairs at the posterior end of the ninth tergite are less numerous than in C. cztromeus, the lateral finger-like processes more slender, and more pointed; the claspers are more highly chitinised; the harpes resemble more closely those of C. grahami than those of C. c2zévoneus; and the proximal ends of the limbs of the aedoeagus are inverted. GoLp Coast: Koforidua, April, 1921 (Dr. J. F. Corson); one male taken in a bungalow, on the wall near a lamp. We have pleasure in dedicating this species to the collector, Dr. J. F. Corson. Culicoides nigeriae, sp.n. MEAsuREMENTS. : Length of body (three females)... ies Bs ao .. 2mm. Length of wing... ae oe not sy a .. O'9mMmM. Greatest breadth of wing ... Sear teat ae re SPORCEE piney Head dark brown. Eyes separated, internal chitinous thickening well developed; the eyes are more widely separated than in C. inornatipennis, tne length of the internal chitinous thickening being about 20m instead of about 13. Proboscis dark brown. Palpi brown, third segment moderately inflated. Aztenna dark brown; segments four to ten from once to once and a half as long as broad. Thorax uniformly dark brown, pollinose, sparsely clothed with dark brown hairs. Scutellum uniformly dark brown; bearing two admedian and two lateral bristles, and a few short hairs. Post- 326 scutellum dark brown. Pleurae dark brown. Wzxgs unspotted; distribution of decumbent hairs similar to that in C. znxornatipennis, but wings rather less hairy. -Halteres darkish brown. Legs rather dark brown, almost unicolourous. Addomen dark brown. Spermathecae two, dark brown and very highly chitinised, sub- spherical to oval, measuring about 35 by 30 on an average; only the very commencement of the duct is chitinised. NIGERIA (Northern provinces): Gimi, Zaria Province, 27th October, 1920 (Dr. W. B. Johnson); ‘collected whilst biting.’ Duchi-n-wai, about forty miles from Zaria, at an elevation of 2,000 ft. approx., 25th November, 1920 (Mr. L. E. B. Pearse), taken in the act of biting. Torin Province, Kaduna river, 21st December, 1920 and 23rd December, 1920; Pategi, 24th December, 1920 (Dr. J. R. C. Stephens); some of the specimens taken on the arm, and undoubtedly biting. Several specimens obtained from each locality, all females. This species, besides being a much darker brown insect, may be distinguished from C. ¢xornatipennis by the following points amongst others: the absence of the characteristic thoracic adornment, the colour of the scutellum and the-colour of the halteres. Culicoides inornatipennis, C., |. and M., var. rudilus, var. nov. A small rufous variety of C. inornatipennis, C., I. and M. Length less than 1 mm., usually about 08 mm.; length of wing about 0'7 mm., and greatest breadth about 0.3 mm. Hypopygium of the male and spermathecae of the female as in C. zxornatipennis ; other morphological characters also similar. Fourth segment of the palp very small, about half the length of the fifth; scutellum bearing three or four stout bristles, one or two central or admedian and two lateral, and a variable number (about half a dozen usually) of smaller hairs. Colouration notably different from that of C. znornatipennis. Head dark brown. Proboscis and palpi pale brown. Antenna with a dark brown torus and lighter brown flagellum segments, bearing pale brown hairs. Thorax unicolorous, light brown or almost golden-brown. Scutellum light brown; post-scutellum darker but not very dark brown. Pleurae brownish-yellow. Halteres pale 327 yellowish-brown. Legs pale brown, almost unicolorous, but with knee spots a slightly deeper yellow-brown. Abdomen dark brown. Pupa. Two pupal pelts were examined, and were found to be indistinguishable from those of C. inornatipennis. They were very small, length about 1°3 mm. GOoLp Coast: Nsawam, May to August, 1920; numerous specimens reared from rotting fibrous material taken from the bases of banana stumps. October, 1920; one specimen reared from material taken from a rot-hole in a silk-cotton tree. Genus DASVYHELEA, Kieff. Dasyhelea fuscipleuris, C., I and M. - In a previous paper, Part IV of this study (1921), this species was described. At that time we possessed only two females, taken in buildings, and had identified neither the early stages nor the habitat. Recently we have reared a large number of specimens, and are able to supplement our previous description by giving an account’ of the pupa. It is interesting to note that all the specimens reared were females and to compare this observation with those previously made on C. clarkei. and C. ertodendroni (1920). Pupa. Length about 2°1 mm., delicately chitinised and rather slender. Respiratory trumpets very long and slender, length about o'6 mm., breadth about 19, raised on small tubercles; the main tracheal trunk is narrow, gives off throughout its length, beginning at its very base, numerous (about fifteen) quite short lateral branches, and ends distally in a cluster of about six short processes. Cephalo- thorax not very strongly chitinised, operculum coarsely shagreened. Anterior marginal tubercle large, conical, bearing a small spine-like hair; dorso-lateral irregularly shaped, bearing two delicate hairs; anterior dorso-median small, bearing two very short, straight hairs; ventro-lateral almost obsolete, bearing two minute hairs, ventro- median represented by a small hair. Dorsum of the thoracic region not infuscated, without tubercles, but with several brownish macules. Abdomen feebly chitinised, of the usual form; tubercles small and poorly chitinised, terminal processes widely divergent. Larva. The larva is of the usual form. Only a single pelt actually correlated with an adult was obtained, and in it no 328 characteristic points could be made out. Unfortunately, it was a practical impossiblity to isolate the larvae of this insect from the materials in which they were living, because the sample contained also numerous other species (Culicoides schultzei, C. similis, C. austeni, C. distinctipennis, Stilobezzia spirogyrae and Dasyhelea ¢ncons picuosa). The single larva reared through to the adult stage was isolated in a small tube containing nothing but a little damp filter paper. The larva buried itself immediately but a day or two later the pupa was observed worming its way to the surface through the mass of filter-paper. The trumpets could not be distinguished with the aid of a hand lens. Two days later a female C. fusczpleuris emerged. The pupal pelt was found on the side of the tube half an inch above the filter-paper, a situation which the pupa must have reached by its own unaided efforts. The larval pelt was found embedded in the filter paper near the spot where the pupa was first observed. GoLp Coast: Accra, April, 1921; numerous specimens (all females) reared from mud taken from the edges of pools and puddles near the station for the Weshiang railway line (Pl. XXI, fig. 2). Dasyhelea nigricans, C., 1. and M. This species was originally described from two males taken in the laboratory at Accra. The halteres of these specimens had yellowish-brown knobs and dark brown stems. More recently we have received from Dr. W. B. Johnson a number of specimens of a species of Dasyhelea taken at Kaduna, Nigeria, in August, 1920, which resembles in every respect Dasy/elea nigricans, excepting that in some the halteres are white, and in others yellow. We conclude, therefore, that in this species the colour of the halteres must be variable. A few of the specimens collected by Dr. Johnson at Kaduna were females, and we are, therefore, able to supplement our previous account by describing the characters of this sex. MEASUREMENTS. Female. Length of body uf; a & es an 3, i regimmt Length of wing et £4 ie is aan ie .. T'omm, Greatest breadth of wing ... an Gs ade ae ... 0°35 mm. 329 The female resembles the male in most respects, but the following points, including the more important differences, may be mentioned. Head eyes separated. Antennae: hairs dark brown, short and scanty ; segments of the flagellum gradually elongating from base to apex in a continuous series, that is without an abrupt change of form between the tenth and eleventh segments; segments four to ten from once to nearly twice as long as broad, segments eleven to fifteen from twice to a little over three times as long as broad, the last segment ending in a stylet; long spines present on all the segments excepting the last five. Thorax: scutellum almost entirely yellowish-brown; armature of bristles and hairs as in the male. Wings rather densely hairy, the hairs extending basally beyond the cross-vein ; bifurcation of the fifth vein at about the same level as the termination of the costa; cell formed by the first and third veins at their junction with the costa larger than in the male. Halteres yellow. Legs: claws small, simple, equal. Addomen clothed with yellowish-brown hairs; spermatheca single, highly chitinised, sub- spherical (diameter about 42), the commencement of the duct chitinised for a considerable distance, about 15“, most strongly at its end of origin from the spermatheca. NIGERIA (Northern provinces): Kaduna, August, 1920 (Dr. W. B. Johnson). Dasyhelea nigeriae, sp.n. MEASUREMENTS. Male. Female. Length of body... ae da sas .. II mm. I'l mm. Length of wing... ae Be a .. o8 mm, o'8 mm. Greatest breadth of wing ... ee, oa 2 07% mm. o°'3 mm. Head dark brown. Eyes separated in both sexes. Clypeus and proboscis dark brown. Palpi dark brown; third segment inflated, in the male especially basally, about the same length as the second ; fourth segment short, about half the length of the third. Awzdennae unusually short, dark brown, with dark brown hairs; in the female segments four to fourteen gradually lengthening towards the apex, the length varying from two-thirds to once the breadth, the last segment rather larger and longer and ending in a conical tip, but without a stylet, in the male segments four to eleven short, those at the base breader. than long and those at the apex sub-spherical, 330 length varying from about two-thirds to once the breadth, last four segments sub-equal, about four times as long as broad, not binodose but exhibiting the usual sculpturing (compare D. flava), the last segment longer and stouter but without a stylet. Zorvax uniformly dark brown, with paler, somewhat yellowish, humeral patches. Scutellum dark brown, bearing in both sexes one central, two admedian, and two lateral bristles and no-short hairs. Post- scutellum dark brown. Pleurae dark brown. Wings hyaline, without spots; clothed with decumbent hairs which do not extend basally beyond the cross-vein. Venation as in D. flava; interspace very small in the male, more well developed in the female. Halteres with dusky orange-coloured knobs and infuscated stems. Legs brown, femora and tibiae dark brown, but lighter coloured in the male than in the female. Claws short, equal, simple; in the male bifid at the tips. Empodium rudimentary, but in comparison with the size of the claws appearing to be larger than usual. Addomen dark brown, venter slightly paler than the dorsum. Spermatheca single, highly chitinised, pyriform (about 42" by 38); chitinised part at the commencement of the duct conical, about 11 long. Hyropycium (fig. 2). Ninth segment: tergite tapering slightly posteriorly, sparsely clothed with strong hairs, posterior margin not Fic. 2. Dasyhbelea nigeriae, sp.n., outlines of male hypopygium, ventral view. a—ninth segment and forceps; b—harpes; c—aedoeagus. (x 375 circa.) 331 notched, and without finger-like lateral processes ; sternite prolonged posteriorly on each side of the middle line into a highly chitinised, pointed, finger-like process. Forceps: side-pieces short and stout, broader apically than basally, rather scantily clothed with moderately long hairs, and bearing on the inner aspect a highly chitinised conical projection; claspers bifid, both parts well developed, pubescent on their basal halves and bearing also a few longer hairs. Harfes: basal portions unequal, highly chitinised ; from the right basal portion, which is the broader, arises a rather lightly chitinised posterior extension (shaped as shown in the figure), the distal end of which is twisted and covered by minute hairs. Aedoeagus broadly-ending, the processes on each side of the horizontal band highly chitinised, finger-like, and curved ventrally at their tips. NIGERIA (Northern provinces): Kaduna, August, 1920 (Dr. W. B. Johnson). This insect resembles D. flava in some respects, but is of an entirely different colour; the hypopygium of the male is characteristic. Dasyhelea boothi, sp. n. MEASUREMENTS. Length of body (one male) ae vas ie a .. T'omm. Length of wing... a im wae a aa ... O08 mm. Greatest breadth of wing ... seg Es coy) oer ... 0°25 mm. Head dark brown. Eyes narrowly separated. Clypeus, proboscis and palpi brown. Third palpal segment not inflated, about as long as the fourth and fifth together, fifth about as long as the fourth. Antennae: torus dark brown, flagellum brown; segments four to eleven sub-spherical to ovoid, length from about once to once and a third the greatest breadth; segments’ twelve to fifteen elongated, sub-equal, but the fourteenth slightly the shortest, the twelfth, thirteenth and fourteenth about three to four times as long as broad, binodose, the fifteenth broader, not ending in a stylet. Thorax dark brown. Scutellum pale brown, slightly darker at the sides; bearing two lateral and four centro-marginal bristles, and one small central hair. Post-scutellum dark brown. Pleurae brown. Wéangs sparsely clothed with hairs which extend between the fourth and fifth veins to the level of the cross-vein; venation as usual. Halteres 332 with brown stems and yellow knobs. Legs uniformly brown; claws small, equal, simple, bifid at the tips. Addomen dark brown. Hyrorycium (fig. 3, a to d). Ninth segment: tergite sparsely clothed with relatively short hairs, tapering distally, posterior margin not notched and bearing double lateral processes; sternite bare, prolonged posteriorly in the middle as a triangular process. Forceps not very highly chitinised or hairy: side-pieces rather short Fre. 3. @to d—Dasyhelea boothi, sp.n., outlines of male hypopygium ; ventral view : a—ninth segment and forceps; c—harpes; d—acdoeagus; b—lateral view. e—Dasyhelea retorta, sp.n., spermatheca. (X 300 circa.) and broad, with a hairy internal apical process; claspers moderately chitinised, with somewhat squared ends, the proximal two-thirds pubescent, bearing three short, stout, hairs about the middle of the inner border. Harpes (fig. 3, 6 and c) moderately well chitinised, almost symmetrical, composed of a large and irregularly shaped basal plate on each side, and a median common posterior extension which is complex, expanded distally, and with a terminal ventral 333 barb-like process. Aedoeagus (fig. 3, 4 and d) very highly chitinised in parts, apparently composed of a pointed median process which is bent ventrally at its tip and in ventral view is pyriform, two short and broad limbs, and external to them somewhat T-shaped extensions. NIGERIA (Cameroons): Victoria, April, 1921 (Dr. L. H. Booth). We have pleasure in dedicating this species to the collector, Dr. L. H. Booth. Dasyhelea retorta, sp. n. MEASUREMENTS. Length of body (one atid txt a ac See ees Gunn. Length of wing... A nC Oe as ae ... O7>E9mMmM. Greatest breadth of wing SA Rs Ost de +c .. 03% mm. Head dark brown. Eyes very narrowly separated dorsally. Clypeus, proboscis and palpi dark brown. The third segment of the palp cylindrical, not inflated, about as long as the fourth and fifth® segments together; the fifth segment shorter than the fourth, widest at its distal end and rounded. Axtennae dark brown, with dark brown hairs and long curved spines on all the segments of the flagellum ; third segment slightly broader than the fourth; segments four to fourteen forming a continuous series, gradually elongating and becoming more flask-shaped, their length varying from about once to once and a half the greatest breadth; the last segment broader and longer, about three times as long as broad, ending in a stylet. Zhorax dark brown with small yellowish humeral patches. Scutellum almost entirely yellow, sides only slightly infuscated ; bearing two lateral and four centro-marginal bristles, and a single central hair. Post-scutellum dark brown. Pleurae brown, lighter than the dorsum. Wesgs with decumbent hairs extending to the base between the fourth and fifth veins. Costa reaching the middle of the anterior border, and terminating beyond the bifurcation of the fifth vein. First and third veins forming a small cell. Halteres with yellow knobs with brown basal infuscation, and brown stems. Legs light brown, with dark knee spots and slight infuscation of the distal tarsal segments; hind femora slightly infuscated in the middle dorsally. Claws simple, equal. Addomen dark brown, venter paler than the dorsum, yellowish pigmentation (which 334 disappears in caustic potash) visible laterally and between the segments. Spermatheca single, highly chitinised, shaped something like a chemical retort (fig. 32); distal portion sub-spherical, length 44, greatest breadth 38”; proximal portion, the chitinised commencement of the duct, curved, arising obliquely, length about 30m, width at its middle about IIp. SIERRA LEONE: Freetown, May, 1920; one female taken about noon upon a window in the Royal Hotel. Genus ATRICHOPOGON, Kieff. Atrichopogon africanum, sp. nov. MEASUREMENTS. Length of body (one female) st 550 aoe 30 .. 4mm. Length of wing =e oe sé ae oes aa sos MEA: Greatest breadth of wing ... on 500 506 ae .. O74 mm. Head dark brown, with dark brown hairs. Eyes broadly contiguous above, bare. Clypeus, proboscis and palpi dark brown. First and fourth palpal segments short, second and third longer, fifth somewhat dilated at its distal end; third segment slightly inflated and furnished with a moderately large sensory pit in its distal third. Axtennae dark brown: segments four to ten short and broad, the tenth sub-spherical; last five segments (eleven to fifteen) elongate, sub-equal, four or five times as long as broad, the fifteenth terminating in a relatively long stylet. Zhorax uniformly dark brown; clothed with a few short, dark-brown hairs. Pleurae dark brown. Scutellum dark brown, but not so dark as the mesonotum, bearing two sub-median and two lateral bristles and a few (about half a dozen) short hairs. Post-scutellum dark brown. Wangs (fig. 4) generally similar to those of A. xanthoaspidium, unspotted Fic. 4. Atrichopogon africanum, sp.n., outline of wing of female. (x go circa.) 335 but slightly infuscated, anteriorly somewhat darker than posteriorly ; anterior veins brownish. Surface uniformly covered with micro- trichia, and bearing decumbent hairs, which, however, are more scanty than in A. xvanthoaspidium, none being present between the branches of the fifth vein or in the anal fold, and only a very few between the lower ramus of the fourth vein and the upper ramus of the fifth. Petiolate portion of the fourth vein about the same length as in A. xanthoaspidium. Walteres with dark brown knobs. Legs almost uniformly brown, but distal segments slightly darker. First tarsal segment of hind legs rather over three times as long as second. Claws simple, equal, about half the length of the fifth tarsal segment. Empodium well developed, as long as the claws. Abdomen dark brown, ventral surface slightly paler. Spermatheca (fig. 5) single, Fic. 5. Atrichopogon africanum, sp.n., spermatheca. (x 375 circa.) large’(length 107, greatest breadth 84); pear-shaped, heavily chitinised ; a short portion (about 15) of the duct, which is narrow, is chitinised. GOLD Coast: Accra, August, 1920, one female taken in the evening upon a window in the laboratory. Atrichopogon elektrophaeum, sp. n. MEAasuREMENTS. Length of body (one female) ay 30 aoe “ta oa. Diamine Length of wing... wes a ~. AS ae APs oe yeni Greatest breadth of wing ... os so oo a .. O§ mm. Head yellowish-brown. Eyes contiguous above. Clypeus, proboscis and palpi yellowish-brown. First segment of the palp 336 ; short; second, fourth and fifth sub-equal, about twice the length of the first ; third rather shorter than the combined lengths of the fourth and fifth segments, slightly inflated in the middle, furnished with a sensory cup opening at the junction of the middle and distal thirds; fifth segment somewhat infuscated and tapering distally. Antennae brown: first segment poorly chitinised, bearing several short hairs; torus rather a light brown colour, sub-spherical, bearing a few short hairs; third segment wider than any of the other basal segments; segments four to ten short and broad, their lengths varying from about three-quarters to a little over once the breadth; segments eleven to fifteen elongate, cylindrical, sub-equal, four or five times as long as broad, the last the longest and ending in a nipple-lke process. All the basal segments of the flagellum bear long, curved, pointed spines. The ratio of the combined lengths of segments three to ten to the combined lengths of segments eleven to fifteen as 06 to 1. Thorax yellowish-brown, unicolorous, clothed with small brown hairs and bearing larger and darker hairs above the wings. Scutellum pale yellow-brown, bearing two sub-median and twe lateral bristles, and about half a dozen small hairs which are dark brown in colour. Post-scutellum yellowish-brown, darker than the scutellum. Pleurae yellowish-brown. Waénxgs with a pale yellowish tint, similar to those of A. xranthoaspidium but with fewer hairs, none being present between the fourth and fifth veins nor between the rami of the fifth vein, and only a row of eight along the false vein in the anal angle. Halteres with pale yellow-brown knobs. Legs almost uniformly yellowish-brown, terminal segments of the tarsi, however, slightly darkened. First tarsal segment on all the legs about three times as long as the second. Claws simple, equal, about half the length of the fifth tarsal segment. Empodium well developed, as long as the claws. Addomen yellowish-brown, rather darker than the thorax; venter paler than the dorsum. Spermatheca single, highly chitinised, oval, large, length 106”, greatest breadth Son; the duct not chitinised. GOLD Coast: Accra, 1920; taken in the evening upon a window in the laboratory. 337 Atrichopogon perfuscum, sp. nov. MEasurEMENTS. Length of body... ane ef is pe oc eS Glen ihine Length of wing... aa a us es ae eof E38 mine Greatest breadth of wing one ae Pas ote ie .-. O'4 mm. Head dark brown, clothed with dark brown hairs. Eyes broadly contiguous above, smooth. Clypeus, proboscis and palpi dark brown. Antennae dark brown; segments four to ten short and broad, the tenth sub-spherical the others broader than long, the length varying from two-thirds to three-quarters the breadth; the last five segments (eleven to fifteen) elongate, about three times as long as broad. Thorax uniformly dark brown. Pleurae dark brown. Scutellum dark brown, but rather lighter coloured than the mesonotum, bearing two sub-median and two lateral bristles and several (six to eight) short hairs. Post-scutellum dark brown. Wings clear, unspotted; venation and decumbent hairs as in A. xanthoaspidium, but fewer hairs present between the rami of the fifth vein and in the anal angle (only two being present in the former situation and one or two in the latter in one of the specimens examined, rather more in the others). Halteres with buff-coloured knobs and dark brown stems. Legs almost uniformly yellowish- brown, but tarsal segments slightly infuscated. Claws equal, about half the length of the fifth tarsal segment, each with a very small sub-apical tooth. Empodium well developed, as long as the claws. Abdomen dark brown. Spermatheca single, highly chitinised, oval; length from 7oz to 87m, greatest breadth from about 57, to 65m, the duct chitinised for only a short distance (about 5) at its commencement. GOLD COAST: Accra, October, 1920; three females collected in the evening upon the windows of the laboratory. Atrichopogon chrysosphaerotum, sp. nov. MEAsuREMENTS. Length of body (two ae I'l5 mm. Length of wing i & o’9 mm. Greatest breadth of wing ... 0°35 mm. Head dark brown, clothed with dark brown hairs. Eyes broadly contiguous above, bare. Clypeus, palpi and proboscis dark brown. 338 First palpal segment short; second, third and fourth longer, sub- equal; fifth rather shorter; third segment moderately inflated and furnished with a well developed sensory pit. Antennae dark brown, segments four to ten short and broad, the tenth sub-spherical, the others broader than long, the length varying from half to about two-thirds the breadth; last five segments (eleven to fifteen) elongate, from two and a half to three times as long as broad, the fifteenth terminating in a relatively large stylet. Thorax uniformly dark brown, scantily clothed with brown hairs. Pleurae dark brown. Scutellum dark brown, bearing two sub-median and two lateral bristles and no short hairs. Post-scutellum dark brown. Wings clear, unspotted, the anterior veins brownish; venation as in A. xanthoaspidium, but first cell larger; decumbent hairs absent, wing surface covered by microtrichia. Halteres with yellow knobs and pale straw-coloured stems. Legs almost uniformly yellowish- brown, but tarsal segments slightly infuscated. Fourth tarsal segments not cordiform; first tarsal segment of hind legs nearly four times as long as second; dorsal hairs on the tarsal segments rather long. Claws equal, apparently simple or with a very minute sub- apical tooth, about half the length of the fifth tarsal segment. Empodium well developed, as long as the claws. Addomen dark brown with a yellowish tint, pigmented with a substance which does not clear in carbolic acid but which is removed by caustic potash. Spermatheca single, highly chitinised, oval; length 57, greatest breadth 46m, only the very commencement (about 4) of the duct is chitinised. GOLD Coast: Accra, November, 1920; a single female, collected in the evening upon a window in the laboratory. Oblogo, May, 1920, reared from rotten wood from a canoe in the river Densu. Atrichopogon homoium, sp. nov. This species, of which we possess only a single female, agrees with the foregoing (A. chrysosphaerotum, sp. nov.) in size, coloura- tion, and apparently in every other particular excepting in the distribution of the hairs on the wings and the scutellum. On the scutellum are four short hairs, two on each side, in addition to the bristles. On the wings are a few decumbent hairs; seven to ten at 339 the tip of the wing, one near the periphery between the rami of the fourth vein, and a few along the distal portion of the anterior ramus of the fourth vein (fig. 6). Yn, U- Tips ae TI OTT rere Fic. 6. Atrichopogon homoium, sp.n., wing of female. (x 120 circa.) The distribution of the hairs on the wings and the scutellum appear to be important specific characters, and therefore, notwith- standing the similarity of this insect to A. chrysosphaerotum and the paucity of our material, we feel compelled to regard it as a separate species. GOLD CoAsT: Oblogo, September, 1920; a single female reared from material taken from a canoe. Genus KEMPIA, Kieff. This genus, originally described by Kieffer (1913) as a sub-genus of Dasyhelea and subsequently raised to generic rank by its author and removed to the Az7zchopogon group, appears to be characterised chiefly by the presence of a well developed empodium on the legs and of pubescence on the eyes, and the absence of the longer hairs from the wings. We have referred the insect described below to the genus Kemfza, notwithstanding the fact that it bears on its wings a few decumbent hairs, because, as stated elsewhere, we are inclined to regard this character as of specific rather than generic value. It may be noted, however, that in the case of Prokempia the absence of the longer wing hairs appears to have been considered by Kieffer sufficient justification for its separation from Dasyhelea. 340 The larvae and pupae resemble those of Forcipomyia. The larvae were found living upon the water lettuce Pistia stratiotes, but were normally seldom seen alive, probably because they were buried in the substance of the plant. The pupae, the posterior portions of which are enveloped by the larval pelts, were found partially embedded in the leaves and between the papillae which cover densely the surface of the leaves. They appeared to be sedentery. The genus, therefore, shows affinities to three different types of the Ceratopogoninae, the characters of the wing linking it to Atrichopogon, those of the eyes to Dasyhelea, and those of the early stages to Forcipomyta. Kem pia ochrosoma, sp. nov. MEasuREMENTS. Length of body... oe 353 re $5 bets <2) gem, Length of wing ae on OAL 436 Sor 506 -, Iagmme: Greatest breadth of wing ... aes SS ae a -s) 0°35 mm. Head straw-yellow, clothed with yellow hairs. Eyes pubescent : in the female contiguous, but with the facets narrowly separated ; in the male separated. Clypeus, palpi and proboscis pale straw- yellow, with similarly coloured hairs. First palpal segment smal!, second and fourth sub-equal, fifth short and somewhat expanded at its distal end, third longer than any of the others, slightly inflated in its middle third, and furnished with a deep sensory cup. Antennae: first segment and torus straw-yellow; in the female, flagellum segments slightly infuscated, especially the more distal ones, segments eleven to fifteen dark» brown; in the male, segments three to eleven pale yellow, segment twelve dark distally, and the last three segments dark brown; whorls of hairs pale straw-yellow. In the female, segments four to ten sub-spherical, the length varying from a little less than to about the same as the breadth, segments eleven to fifteen elongate, from twice to three times as long as broad, the last segment terminating in a stylet. In the male, the last three segments elongate, sub-equal, the thirteenth slightly the longest and the fifteenth terminating in a stylet. Thorax uniformly coloured, ochraceous, almost the same hue as the head; clothed with short, curved, yellow hairs dorsally, and with a few longer, spine-like, 341 yellow hairs, posteriorly. Pleurae ochraceous. Scutellum slightly paler yellow than the mesonotum, bearing two admedian and two lateral bristles and a few (ten in the female, eight in the male) short hairs. Post-scutellum ochraceous. Wzxgs unspotted, the anterior veins yellowish ; venation as shown in the figure (fig. 7). Decumbent “Fic. 7- Kempia ochrosoma, sp.n., wing of female. (x 90 circa.) hairs scanty: in the female, limited to the distal third of the wing and a few near the posterior margin; in the male, almost entirely wanting. Surface of the wing closely covered with microtrichia. Halteres with pale straw-yellow stems and white knobs. Legs almost uniformly pale straw-yellow; unarmed. Claws in both sexes equal, about half the length of the fifth tarsal segment ; in the female, each with a very small, sub-apical tooth, in the male, deeply bifurcated at the tips. EEmpodium well developed, hairy; at least as long as the claws. Abdomen ochraceous, rather paler yellow than the thorax. Hypopygium of the male of a similar colour and feebly chitinised. Spermatheca single, rather feebly chitinised, obovate and very large; length about 230, greatest breadth about 150z, the duct chitinised at its commencement for a short distance (about 10“). The abdomen, and also the thorax and the knobs of the halteres, containing a yellowish pigment which is not cleared by carbolic acid but which dissolves in caustic potash. Hyrorycium (fig. 8 a and b). Nenth segment feebly chitinised ; tergite rather short and hairy, posterior margin rounded, without lateral, finger-like processes. Forceps: side-pieces normal, feebly chitinised; claspers also feebly chitinised, about as long as the 342 side-pieces, tapering distally, hairy almost to their tips. MHarpes apparently absent. Aedoeagus feebly chitinised, a large ventral median structure with a central and two lateral processes ; when fully d Fic. 8. Kempia ochrosoma, sp.n. a and b—Male hypopygium, ventral view of aedoeagus ; c—posterior end of abdomen of pupa; d—dorsal gill-like tubercle of cephalo- thorax of pupa. (a, b, and c x 250 circa; d X 400 circa.) — expanded as shown in fig 8 a, but perhaps more frequently appearing as in fig. 84. We are not able at present to suggest what are the homologies of the median structures. Pupa. Length about 21 mm., feebly chitinised; the posterior half of the abdomen (from the fifth segment) enclosed within the larval pelt, which is much shrivelled and discoloured. Integument irregularly covered with rather sparsely scattered spicules. Cephalo- thorax relatively large and broad, somewhat of the Forcipomyia type but with no dorsal extension over the middle of the first abdominal segment. Respiratory trumpets almost smooth, short and nearly straight, length a little less than 0°2 mm. ; they arise from 343 small tubercles and are without stalks. Main tracheal trunk does not give off any lateral branches, and ends distally in a double row of rather long, blunt processes. Operculum feebly chitinised, sparsely spiculated ; at its posterior angle is a small tubercle covered with long spicules. Anterior marginal tubercle an irregularly conical elevation covered by long spicules, and bearing a long bristle ; anterior dorsal situated posterior and external to the anterior marginal and anterior to the base of the trumpet, a small tubercle bearing a rather long hair; dorso-lateral apparently unarmed, except by spicules, ventro-lateral apparently absent or obsolete; ventro-median very small, bearing a short hair. Dorsal tubercles remarkable, six on each side, each giving rise to a delicate, branching, dendritic process resembling a sort of gill (fig. 8d); these processes are well developed on all the tubercles excepting the small admedian pair situated a little anterior to the most posterior pair; two of these tubercles are armed, the two situated most anteriorly and internally, and bear long spines. Addomen directed straight backwards, tapering rather rapidly, the last segment terminating on each side in an almost straight process which is slightly dilated at its distal end, and is covered with long spicules directed anteriorly, which no doubt function as hold-fasts (fig. 8c). The segments, which are enclosed within the larval pelt, bear only rudimentary tubercles; on the other segments (one to four) are a pair of dorsal admedian, and a pair of lateral tubercles similar to those on the dorsum of the cephalo-thorax—that is, dendritic processes resembling gills—the dorsal ones bear large spines. LarvA. Length about 3 mm. when fully grown, brownish in colour. Mead moderately chitinised, more or less conical, bearing a few relatively large hairs or bristles. Horn-like appendages on the dorsurn (which are connected with the distal ends of the antennae of the adult) small, narrow, about 0°06 mm. long, straight, arising from quite small tubercles. Eyes large, bilobed. Mandibles densely chitinised, black, terminating in three or four teeth. Hypopharyngeal! sclerite very highly chitinised, the posterior part armed on each side with a comb-like row of about ten short, pointed teeth. Body composed of twelve visible segments; cuticle covered with coarse spicules. Armature of bristles, spines, etc., modified at the anterior and posterior ends, but general arrangement as follows. 344 A pair of finger-like, admedian dorsal tubercles, bearing long spines, which are mere or less barbed at their bases; three pairs of very long (about o'8 mm.) and delicate dorso-lateral processes; four pairs of ventral spines, two central, the one admedian and the other ventro- lateral, and two postero-marginal, both ventro-lateral, these spines are relatively short and are freely barbed. The long dorso-lateral processes are the most conspicuous features of the larva, they bear lateral spines or bristles, are directed backwards, and trail behind the larva. The pro-thoracic pseudopods are partially fused, highly spiculated, each armed with a group of about ten large, well developed hooks, and anteriorly with numerous small hooks. The anal pseudopods are armed with similar large hooks arranged in two transverse rows. GOLD Coast: Oblogo, February to March, 1921; reared from plants of the water lettuce (Pistza stratiotes) taken from a swampy pool, and from backwaters of the river Densu (Pl. XXI, fig. 1). Genus MONOHELEA, Kieffer. This genus is stated by its author to possess the characters of Stilobezzia, Kieff., but the petiole of the fourth longitudinal vein is very short, the fourth tarsal segment is long and cylindrical, the claws of the fore and middle legs of the female are simple, equal; two-thirds the length of the last tarsal segment, and the claws of the hind legs single, longer than the last tarsal segment. The species described below, of which at present we possess only a single female, appears to belong to this genus, although not conforming exactly to the generic description given above (for example, the claws of the fore and middle legs are not simple). The insect superficially resembles S/zlobezzia, and when at rest holds its wings in a manner similar to S. spzrogyrae, that is, diverging slightly, and not folded one on top of the other on the dorsum of the abdomen. Monohelea htoraurea, sp. n. MEasuREMENTS. Length of body (one female) Ae a ee Se nos ™ 73 TH Length of wing 30 b eis 5 ie Ate .. T'Omm. Greatest breadth of wing ... a aad = he ae .. O74 mm. Head: occiput dark grey to brown, clothed with dark brown hairs. Eyes smooth, contiguous dorsally. Clypeus and proboscis 345 dark brown. Palpi dark brown, third segment slightly inflated and furnished with a sensory cup; fifth segment rather long and narrow, with a terminal group of four hairs. Axdennae darkish brown: first segment rather large, dark brown, bearing a few dark hairs; torus dark brown, bearing a few dark hairs; flagellum segments paler, gradually elongating and deepening in colour towards the distal end of the antenna; fourth to tenth segments sub-cylindrical, from about one and a third times to twice as long as broad; segments eleven to fifteen elongate, from-about three to a little over four times as long as broad, the last segment being the largest and the longest and tapering at its extremity, but not terminating in a definite stylet. Whorls of hairs small and the constituent hairs short; there is a hair just before the tip of the last segment. Zhorvax: dorsum dark grey with many dark brown spots and patches, and with the antero- i), nergy ny Fic. 9. Monobelea litoraurea, sp.n., wing of female. (x 105 circa.) lateral angles brownish-yellow. The arrangement of the spots and patches is somewhat like that on the thorax of Culicoides schultzet. The mesenotum is clothed with rather short, dark hairs. Pleurae dark brown. Scutellum darkish brown, but not so dark as the mesonotum, paler in the middle than at the sides, bearing two admedian and two lateral bristles and a few (six) short hairs. Post- scutellum dark brown. Waéngs grey, with large white patches; the arrangement of the patches and the venation as shown in fig. 9. Lower ramus of fourth vein obsolete at the proximal end. The surface of the wing is covered by microtrichia, but without larger decumbent hairs. Halteres pale, with white knobs. Legs brown, femora and tibiae more or less infuscated. Fore and middle 346 femora brown, somewhat infuscated, especially at their apices, not swollen and without strong spines; hind femora uniformly very dark brown, slightly swollen, without strong spines. Fore and middle tibiae brown, infuscated; hind tibiae uniformly very dark brown, somewhat swollen. Tarsal segments brown, paler than the proximal segments: fore and middle tarsi without regularly arranged rows of spines, fore tarsi with a weli developed black spine at the apex, the base, and sometimes the middle of the first segment and somewhat smaller but similar spines at the apex of the second and third segments, middle tarsi with several similar but less well defined spines, on the first segment and at the apex of the second and third segments, hind tarsi (fig. 10@) with a regularly arranged ventral Fic. 10. Monobelea litoraurea, sp.n. a—hind tarsus of female (x 190 circa); b—spermatheca of female (x 375 circa). row of small spines on the first segment, and with large black spines, one at the apex and one at the base of the first segment, two at the apex of the second segment, one at the apex of the third segment, and two at the apex of the fourth segment. Fourth tarsal segment on all the legs cylindrical. Claws on the fore and middle legs small, about half the length of the fifth tarsal segment, equal, each with a small basal tooth and a bifid extremity; claws on the hind legs single, with a very long tooth (as long as the fourth and fifth tarsal segments together), and a short tooth apparently fused with the large tooth at the base. Empodium rudimentary. Aédomen very dark brown, excepting at its extreme proximal end, where it is yellowish-brown; distal end almost black. Dorsal surface very sparsely clothed with short, dark hairs, which are almost entirely restricted to the sides. Spermathecae (fig. 106) two, unequal, 347 highly chitinised, pyriform (measurements about 80” by 55m, and 65 by 55m); the commencement of the duct, which is narrow (about 4), is chitinised for some distance (about 20). GOLD Coast: Accra, 27th March, 1921; collected on a window in the labcratory at 6 p.m. Genus EUKRAIOHELEA, nov. The two following species show kinship to the genus S/dlodezzia, Kieff., the chief generic characters of which are, according to Kieffer (1919)—wings glabrous, the first and third veins forming two radial cells of which the second is the longer, the fourth vein petiolate, the fourth tarsal segment cordiform in both sexes, and the claws long, simple, and very unequal in the female, short and equal in the male. Thy possess the above characters excepting that the first radial cell of the wing is obsolete, the first and third veins forming only a single large cell, but they show certain other divergencies from earlier descriptions given by Kieffer (1917), for example, the fore femora are armed but not swollen, and the hind tibiae bear spines. Moreover, the hypopygium of the males, whilst closely resembling one another, diverge from the type found in Stilobezzia spirogyrae. These morphological differences are, in our opinion, of such a nature as to necessitate the separation of these two species from the genus Séi/odezzia, and, therefore, notwithstanding their close similarity in other respects to species of that genus, we propose to regard them as belonging to a new genus, for which we suggest the name Eukraiohelea. Lukraiohelea africana, sp. n. MEasuREMENTS. Male. Female. Length of body... ve tet “a << wns) mM. I'¢ mm. Length of wing... a aa ae .. 2mm. I°3 mm. Greatest breadth of wing ... ee Seared tuacas) Pp Otd, MM. o'5 mm. The ground colour of this insect is olive green, on which are superimposed the brown markings. Head brown. Eyes smooth; narrowly separated in the female, more widely in the male. Clypeus and proboscis dark brown. Palpi dark brown, moderately long, 348 the third segment slightly inflated distally and furnished with a shallow sensory cup near its apex. Mouth-parts well developed in the female. Awtennae: first segment small; torus yellowish-brown, sub-globular, very large in the male, bearing a few short hairs; flagellum pale brown proximally, the last three segments in the male and five in the female dark brown. Plume of the male moderately well developed, pale brown; hairs of the flagellum of the female very short, brown. In the male, the last three segments of the flagellum elongated, the thirteenth and fourteenth about five and twelve times as long as broad respectively, the fifteenth much longer, about thirty times as long as broad, and ending in a short, stout process. In the female, segments four to ten pale brown proximally, dark brown distally, and segments eleven to fifteen all dark brown; segments four to ten sub-cylindrical, rather wider in the middle than at either end, sub-equal, about three times as long as broad; Fic. 11. Eukraiohelea africana, sp.n., venation of wing of male. (x go circa.) segments eleven to fourteen elongated, about ten times as long as broad ; the last segment much longer, about twenty times as long as broad, ending in a short, stout process. Zovax blue-green, with brown infuscation on the dorsum; hairs large, scanty, dark brown. Pleurae blue-green; a dark brown spot on the coxae of the fore and middle legs. Thoracic pits absent. Scutellum blue-green in the middle, and brown, with a blue-green tint, at the sides; bearing two admedian and two lateral bristles and a few (two to four, but often absent in the male) short hairs. Post-scutellum dark brown with a greenish-blue tint. W2zxgs unspotted, without decumbent hairs. Wing surface covered by microtrichia. Venation as shown in the figure (fig. 11); first radial cell obsolete; in the female, the fourth 349 vein forks distally to the fifth, in the male, they both fork at about the same level. Fringe very short on the distal part of the wing. Halteres with greenish-blue knobs; stalks almost white, bases of the knobs infuscated. Legs pale brown, almost colourless, with dark knee spots; the infuscation on the hind legs extending beyond the knee nearly half way down the tibia. Fore femora armed with two short, stout, ventral spines, middle and hind femora unarmed ; hind femora slightly swollen, fore and middle normal. Fore tibiae with the usual long apical spine; hind tibiae with a ventral row of spines, three long ones on the middle third and four short ones more distally. On all the legs the first tarsal segment at least twice as long as the second, third small and almost cordiform, fourth very small and strongly cordiform, fifth slightly infuscated, about as long as the second and longer than the third and fourth together. Rows of bulbous spines present on the first and second tarsal segments of all the legs; a stout basal spine on the first tarsal segment of the middle and hind legs; on the proximal half of the fifth tarsal segment of the fore and middle legs are two pairs of stout spines, on the hind legs a single pair. Claws alike on all the legs: in the female, single, with a long tooth (as long as the fifth tarsal segment), and a tooth about half this length apparently fused with it at the base, which is, moreover, somewhat extended as a process; in the male, shorter (about two-thirds the length of the fifth tarsai segment), bifid at the tips, composed of two equal parts fused at their bases. Empodium absent. Addomen green, with brown infuscation (which is most marked in the female) at the sides and posterior dorsal margins of the segments. First segment with conspicuous lateral tufts of hairs. Spermathecae two, highly chitinised, pyriform (about 46 by 35); the commencement of the duct chitinised for a short distance (about 7) only. Hypopycium (fig. 12). Ninth segment: tergite rather feebly chitinised, moderately hairy; posterior margin broad, not cleft, without finger-like lateral processes; apical lobe-like processes moderately well developed, hairy ; sternite apparently not excavated in the middle posteriorly. Forceps: side-pieces moderately long and hairy, distal ends infuscated ; claspers rather broad, not strongly chitinised, tips infuscated, clothed with very delicate minute hairs and bearing a few longer hairs. Harpes in form somewhat 55o resembling those of species of Dasyhelea, but with the distal extensions bilateral and separate. Basal portions irregularly shaped, broad laterally; posterior extensions long, lath-like processes directed almost straight backwards, with bluntly pointed tips, reaching posteriorly as far as, or a little further than the margin Fic. 12. Eukraiobelea africana, sp.n., outline of male hypopygium, ventral view. (x 375 circa.) of the ninth tergite. Aedoeagus apparently composed of two separate chitinised lateral rods corresponding to the limbs of the arch in other genera, and a delicate membranous portion enclosing them and prolonged posteriorly in the middle line as a conical process. Ventral wall not spiculated. GOLD Coast: Swamp between Koforidua and Tafo, a little north of Accra, April, 1921; Weshiang, June, 1921; reared from plants of the water-weed Pistia stratzotes. 351 Eukratohelea versicolor, sp. n. MEasurEMENTs. Male. Female. Length of body oe te hen RRS Seeger akan’, 1°3 mm. Length of wing... ue eee ate “ieromm: I'l mm. Greatest breadth of wing ... san a ... 0°73 mm. o*4 mm. This insect resembles the last species, but the ground colour is white instead of blue-green. In the following account, only the more important points of difference between it and Eukratohelea africana will be given. Head dark brown; occiput dark brown in the middle, paler at the periphery; median occipital hairs long, dark brown. Eyes during life metallic green; smooth, separated in both sexes. Proboscis dark brown, well developed in the female. Palpi dark brown, as in &. africana. Antennae: torus yellowish-brown ; flagellum very pale brown at the base, last five segments in the female and three in the male completely dark brown, apical portions of the fourth to the tenth segments in the female infuscated. Plume in the male moderately well developed, pale brown, hairs in the female short, pale brown. In the male, the last three segments elongated, about five, twelve, and twenty-two times as long as broad respectively, the last segment tapering to a conical tip. In the female, segments four to ten as in £. africana; segments eleven to fourteen elongate, sub-equal, about eight or nine times as long as broad ; segment fifteen longer, about twelve times as long as broad, ending in a conical tip. Thorax: ground colour white; anterior half of the dorsum almost entirely dark brown in the male, in the female more or less infuscated and dark brown, but with median pale areas on each side of the middle line; in front of the scutellum is a triangular dark brown mark with its base directed posteriorly. Pleurae white. Over the bases of the coxae is a small, oval, dark brown patch. Scutellum dark brown, armature of bristles and hairs as in EZ. africana Post-scutellum dark brown. Wings as in E. africana. Halteres white. Legs: ground colour white; distal half of the hind femora and apical sixth of the hind tibiae dark brown; fifth tarsal segments hardly infuscated. Coxae infuscated. Femora shaped and armed as in E. africana. Tibiae shaped and armed as in £. africana, but hind tibiae bearing only three long spines. Tarsal segments as in £. africana; spines 352 on the fifth segments, however, less well developed, arrangement in the female as in £. africana, in the male only one pair present on all the legs. Claws as in E. africana. Abdomen white, with dark brown markings arranged as follows: small lateral patches on the first, fourth, and seventh segments, large dorso-lateral patches, reaching almost to the middle line dorsally, on the second, third, fifth and sixth segments. Lateral hair tufts on the first segment not so prominent as in £. aj7icana. Spermathecae similar to those of E. africana; two, highly chitinised, pyriform (about 35” by 30), the commencement of the duct chitinised for a short distance (about 4) only. Hyrorycium (fig. 13). Hypopygium darkish brown, rather small. Nenth segment: tergite moderately hairy, tapering slightly distally, posterior margin nearly straight, with a trace of a median Fic. 13. Eukraiobelea versicolor, sp.n., outline of male hypopygium, ventral view. (x 375 circa.) cleft, but without lateral finger-like processes; apical lobe-like processes well developed, hairy; sternite apparently prolonged posteriorly as a delicate, median, cone-shaped process. Forceps: side-pieces rather short and stout, moderately hairy; claspers long, stout, feebly chitinised, especially at their distal ends, entirely 353 covered by very delicate minute hairs and bearing at the posterior extremity a few rather larger hairs. Harpes similar to those of E. africana, but basal portions more expanded laterally, and posterior extensions longer and bent sharply in an anterior direction at about their middles. Aedoeagus somewhat similar to that of E. africana, delicate membranous part apparently with large lateral folds; ventral wall not spiculated. GOLD Coast: Swamp between Koforidua and Tafo, a little north of Accra, April, 1921; reared from plants of the water-weed Pistia strattotes. Genus SCHIZODACTYLUS, nov. This genus is allied to Xylocry pra, Kieff., and Xenohelea, Kieff., genera which have been separated by Kieffer (1917) from Sphaeromias (Stephens), Curtis, and Palpomyza, Mergele, by the characters of the -fourth tarsal segments, which are cylindrical in both sexes, and the antennae of the males, only the last three segments of which are elongated. From the former it may be distinguished by the facts that the eyes in the male are separated and the body is not squat; from the latter by the fact that the claws of the female are equal. The chief generic characters are as follows :—Eyes smooth, separated in both sexes ; widely in the male, narrowly in the female; last three segments of the antenna of the male elongate; wings covered by microtrichia but without longer decumbent hairs, costa reaching beyond the middle of the wing, first and third veins forming two radial cells, the distal of which is the longer, cross-vein not very oblique and not twice as long as the base of the cubitus, fourth vein sessile; femora armed, fourth tarsal segments cylindrical, claws in the female long and equal, those on the fore legs with long basal barbs, empodium rudimentary. Schizodactylus telmatoscopus, sp. n. MEASUREMENTS. Length of body (two males) sa Hs i ose .. 28mm. Length of wing... os Fi ig a3 ve Pulp tg yp Greatest breadth of wing ... = ses Sad ae ... O05 mm. Head dark brown, large, wider than the thorax. Eyes glabrous, widely separated. Proboscis dark brown, very short. Palpi 354 brown, very small; third segment not inflated, sensory pit small or rudimentary. Aw/ennae rather dark brown, especially the torus and the three terminal segments. First segment without hairs; torus sub-spherical, very dark; third segment rather larger than the following segments; fourth to twelfth segments almost cylindrical, from one and a third to two and a half times as long as broad, sharply separated from one another; last three segments elongated, the ‘fifteenth being the longest and not ending in a stylet. Hairs not very long, pale coloured, arranged somewhat irregularly and not forming a single whorl. Tovar uniformly dark brown; small pro-thoracic Icbes present; no tubercle on the front margin of the thorax in the middle; dorsum almost devoid of bristles, and without either anterior or posterior pit-like depressions. Scutellum dark brown, bearing a few (twelve to fourteen) hairs, but no large bristles. Post-scutellum dark brown. Pleurae dark brown. Wangs unspotted and without long decumbent hairs; surface closely covered with minute upright spicules; fringe short; stronger hairs on costa scanty. Venation as shown in the figure (fig. 14); first cell Fic. 14. Schizodactylus telmatoscopus, sp.n., venation of wing of male. (x 70 circa.) rather large and long. Halteres with pale brown knobs. Legs: femora and tibiae dark brown; first four tarsal segments pale coloured with slightly infuscated apices, last segment entirely dark. Trochanters with small, paired, curved spines. Femora not unusually broad, bearing on all the legs two or three stout, short, dark spines on the under surface near the apex. First tarsal segment about twice the length of the second on all the legs; last segment rather elongated, fourth not cordiform. Regular rows of small spines are situated ventrally on the first tarsal segments of 355 the middle legs and on the first and second tarsal segments of the hind legs; those on the first tarsal segments of the hind legs arranged in a double row. Apically the fore tibiae bear a long, pale-coloured, ventral spine, the middle tibiae a strong, dark-coloured spine, and the hind tibiae the usual double row of bristles. First four tarsal segments of the middle and hind legs with a pair of strong, dark spines apically, those on the fourth segments being more slender. Claws about half the length of the fifth tarsal segment, equal, simple, but with bifid ends; empodium rudimentary. Abdomen dark brown; venter slightly paler than the dorsum. Hypopycium (fig. 15). Dark brown, well chitinised, relatively rather small. Ninth segment: tergite bearing few bristles, terminating distally on each side in a large hairy process bearing two relatively long bristles; sternite deeply excavated in the middle, a Fic. 15. Schizodactylus telmatoscopus, sp.n., male hypopygium. a—dorsal view ; b, c and d, ventral views. a—ninth tergite (small hairs not shown); b4—harpes; c—aedoeagus ; d—forceps (hair on side piece not shown). (X 150 circa.) reduced to a narrow band of chitin. Forceps: side-pieces rather narrow, hairs not very long; claspers short, bearing at the distal end a small claw. Harfes fused into a median, strongly chitinised rod ; proximal extremity bifurcated, each half with two processes; distal end expanded, rounded in ventral view but slightly spoon-shaped in lateral view, not highly chitinised, closely covered with minute hairs. Aedoeagus: Y-shaped, chitinised portion with lateral flaps of spiculated membrane on each side; distal end broad, with a small 356 spine at each side, and an irregularly chitinised fringe extending anteriorly ; ventral wall between the limbs of the Y well chitinised, spiculated; membrane connecting the aedoeagus with the ninth sternite with a very few spicules at its distal border only. FEMALE. The following morphological details of the female were made out in a specimen extracted from a pupa of the same sample, and which was apparently identical with those of the males. Head: eyes narrowly separated. Palpi longer than in the male, and segments relatively more slender; first segment small, second, third, and fifth longer and sub-equal, fourth somewhat shorter. Third palpal segment not inflated, without a definite sensory cup but with a slight anterior depression from which arise a number of sensory hairs, the extremities of which are only slightly dilated. Antennae : first segment bearing four hairs; torus somewhat pyriform and bearing a considerable number of short hairs; segments four to ten cylindrical, from about twice to three times as long as broad; segments eleven to fifteen elongate, sub-equal, about six times as long as broad ; last segment not terminating ina stylet. Thorax: scutellum bearing numerous (about thirty-three) long bristles of somewhat unequal sizes and arranged more or less in three rows in the middle and in two rows laterally. Legs: fore femora with ventral row of twelve to fourteen short, stout, dark spines on its apical half; middle and hind femora with about a dozen similar spines, some of those situated nearest the apex being paired. Fore tibiae with a long, feebly chitinised, ventral spine apically; middle tibiae with short, dark, strongly chitinised spine in a similar position. Fifth tarsal segment of all legs with two pairs of stout, dark spines at the base; middle legs, and hind legs less distinctly, with a pair of somewhat similar spines at the apex of the first, second, and third tarsal segments. Double row of small spines on the first tarsal segment of the hind legs complete. Claws long, equal, almost as long as the fifth tarsal segment; those of the fore legs (fig. 16) with a rather large basal barb, which is not present on the other legs. Empodium rudimentary. Aédomen: spermathecae two, highly chitinised, oval; length 123 to 133, greatest breadth 1064 to 1214; only the very commencement (about 4) of the duct chitinised. Pupa. Length about 4mm. to 5mm. Form similar to that of Culicoides, and, therefore, the description will be given on the same 357 lines as was done in the case of that Genus. The pupa is very dark coloured and highly chitinised, especially at the anterior part of the cephalo-thorax. Respiratory trumpets short, broad, and straight, arising from rather depressed tubercles and without definite stalk; length about 260, breadth in the middle about 654. The main tracheal trunk is very broad, straight, without lateral branches, at its distal end terminating in a semi-circular, fan-like arrangement of about fifteen short processes. Cephalo-thorax. Anterior marginal tubercle small, bearing a small bristle; anterior dorsal small, bearing a small bristle; dorso-lateral small, bearing two bristles; ventro-lateral small, bearing two or three short bristles; ventro- median obsolete, represented by two small hairs. Dorsal tubercles Fic. 16. Schizodactylus telmatoscopus, sp.n., fifth tarsal segment and claws of foreleg of female. practically obsolete: anterior represented by two bristles slightly separated from cne another; posterior by a single bristle; and lateral by a bristle and a socket-like mark, apparently unarmed, a little external to it. The dorsum is highly chitinised and very dark, the integument closely covered by small dark granulations; there is a transverse band of dark patches on the anterior third similar to that seen on many species of the Genus Dasyhelea. The postero-dorsal tubercle apparently obsolete. The posterior margin of the dorsum is rounded, not prolonged backwards as a median process. Abdomen: first segment, very short, second, long and broad; third to eighth, sub-equal but, tapering towards the posterior extremity, 358 almost square; ninth, short and small, with two short, sharply- pointed, divergent terminal processes. There are a few dark spots on the integument of most of the segments: dorsally, two sub-lateral anterior, two admedian central, and a central a little farther back; ventrally, two sub-lateral anterior, and a central somewhat heart- shaped spot between and behind them. The tubercles are all small, and bear only small spines or hairs. The following may be distinguished on each side of a typical segment. Dorsal tubercles: antero-submarginal, two, very small, the outer being situated almost laterally; postero-marginal, three, small, the inner one situated posterior to the inner antero-submarginal tubercle, the outer two contiguous, situated almost laterally. Lateral tubercles: antero- submarginal, single, very small; postero-marginal, two, rather larger than the other tubercles, situated close together and rather more anteriorly than the corresponding dorsal and ventral tubercles, the dorsal one appears to be composed of two fused tubercles and bears a small spine and a short hair. Ventral tubercles: three, small, postero-marginal in position and situated almost laterally ; the inner two are the larger and are contiguous, the outer one is very small and is situated slightly more anteriorly than the others. GoLtp Coast: Accra, December, 1920; numerous pupae (of which only two, males, hatched), found in puddles of dirty water near a stand-pipe beside the Weshiang railway line, about two and a half miles from Accra. Genus SPHAEROMIAS (Stephens), Curtis. This genus includes those midges in which the eyes are bare, the wings bare except for minute, point-like hairs visible only with a microscope, third vein extending beyond the middle of the wing, second radial cell longer than the first, fourth vein forking almost under the cross-vein, femora unarmed and not swollen, fourth tarsal segments cordiform, fifth not swollen, empodium absent or rudimentary. It is apparently the same as the Genus /ohannsenomyia erected by Malloch to include those species, which he had previously included in /ohannseniella, ‘ which have the media furcate proximad to the cross-vein.’ One male of a single species of this genus was obtained near Accra. 359 Sphaeromias litoraurea, sp. nov. MEASUREMENTS. Length of body (one male) Bec bac oa 406 wes MO" Length of wing... Af fe 263 if ane scold eae van, Greatest breadth of wing ... e ve “pr: oe ... O'4 mm. Head dark brown. Eyes widely separated, smooth. Clypeus and proboscis dark brown. Palpi dark brown: all five palpal segments short, but the third and the fifth rather longer than the fourth ; third palpal segment slightly longer than broad, not inflated, with a few long, knob-ended sensory hairs on its inner side anteriorly which arise from a very shallow depression. Axtennae dark brown: plume poorly developed, composed of relatively few and short hairs which are not arranged in distinct whorls; segments four to eleven progressively longer, from once and a half to a little over twice as long as broad; segment twelve about three times as long as broad ; segments thirteen to fifteen longer, sub-equal, bearing only short hairs, the terminal segment tapering distally but not ending in a stylet. Thorax uniformly dark brown. Pleurae dark brown. Scutellum dark brown, bearing two admedian, two sub-median, and two lateral bristles. Post-scutellum dark brown. Wéxgs clear, unspotted, the anterior veins brownish. Third vein extending some distance beyond the middle of the wing; two radial cells, both well formed, the first rather large, rectangular, the second longer than the first; fourth vein bifurcated, sessile, the bifurcation taking place a little proximal to the cross-vein. Wing surface covered by micro- trichia but otherwise bare, without decumbent hairs. Halteres with dark brown knobs. Legs almost uniformly brown, but with indications of darker knee-spots and with the tarsal segments slightly infuscated. Trochanters with usual pair of stout, curved spines. Femora unarmed, not swollen. First tarsal segments much longer than second, fourth cordiform. First and second tarsal segments of hind legs with conspicuous ventro-lateral row of small spines. Tibiae of fore legs with a long ventral spine at the apex. Tibiae and first two tarsal segments of middle legs with small, paired, apical spines; first tarsal segment bears also several similar spines on its ventral border. Claws equal, about half the length of the fifth tarsal segment, bifid. Empodium absent. Aédomen dark brown. 360 Hyropycium (fig. 17). Ninth segment: sternite reduced to a narrow strip of chitin; tergite not highly chitinised, bearing apparently only two long bristles dorsally near its middle, the chitinisation of the tergite interrupted a little posterior to them, the posterior margin with a short hairy process on each side, the ventral surface thickly covered by short hairs, the lobe-like processes well developed, covered by short hairs, and bearing one or two longer bristles. Forceps: side-pieces rather long and narrow; claspers short, terminating in strong, pointed hooks, basal two-thirds hairy. s d Fic. 17. Sphaeromias litoraurea, sp.n., outlines of male hypopygium. a, 6, and c— ventral views; d—dorsal view. a—forceps (bristles and hairs of side-piece not shown) ; b—aedoeagus; c—harpes; d—ninth tergite (small surface hairs not shown). asia a Harpes dark brown, highly chitinised, with a double dorso-ventral curve; distal extremities fused to form a blunt, rather broad, process. Aedoeagus conical, proximal portions of the limbs narrow and highly chitinised, distal portions less highly chitinised; distal extremity broad, with a slight ventral lip; ventrai wall slightly | chitinised to about the level of the middles of the highly chitinised portions of the limbs; membrane joining the aedoeagus to the ninth sternite not spiculated. GOLD Coast: Odorkor, a small village near Accra, November, 1920; one male, obtained from a drain situated near a stand-pipe. Weshiang, near Accra, June, 1921; one male, reared from plants of the water-weed Pistia stratiotes, taken from the river Densu. 361 Genus BEZZIA, Kieff. The chief characters of this genus, according to Kieffer (1919), are—eyes glabrous, the last three or four antennal segments in the male elongated, the wings bare or covered by microscopic setae, the first and third veins not united by a cross-vein and forming a single cell, the bifurcation of the fourth vein scarcely proximal to or under the cross-vein, the femora on the fore legs at least armed with one or more ventral spines in both sexes, the fourth tarsal segment cordiform, the fifth unarmed in both sexes, and the claws small, not half the length of the fifth tarsal segment, simple in both sexes, or sometimes with a small median tooth in the female. Up to the present we have not collected at Accra any specimens referable to this genus, but we have received from Lagos a single female of one species, and of this a description is given here. Bezzia foyt, sp. n. MEASUREMENTS. - Length of body (one fone) er anthro cre oe .. 21mm. Length of wing... ae as ue mas .. I°4mm. Greatest breadth of wing ... m. a aN =e .. O'5 mm. Head dark brown, clothed with rather short dark brown hairs. Eyes smooth, narrowly separated. Clypeus, proboscis and palpi dark brown. Mouth-parts well developed, mandibles particularly highly chitinised, and bearing strong teeth on their inner edges. First palpal segment small, second short and broad, third, fourth, and fifth sub-equal, about twice as long as broad; the third segment scarcely at all inflated and without a sensory cup, but bearing a patch of sensory hairs on its inner aspect; the fifth segment pyriform, its distal end broad and rounded, bearing a few rather long hairs. Antennae dark brown, the terminal segments rather paler at their bases; hairs short, dark brown. First segment small; torus sub-spherical, bearing numerous short hairs; third rather longer than the fourth, with a short stem; segments four to ten oval, slightly constricted at their bases, their lengths varying from once and a half to twice the greatest breadth; segments eleven to fifteen elongate, about three or four times as long as broad, the last segment the longest and ending in a conical tip without a stylet. Zhorax 362 uniformly very dark brown, clothed with short, dark brown hairs, and bearing above the wing bases a few longer, strong hairs. Pleurae dark brown. Scutellum dark brown, not so dark as the dorsum; bearing two lateral and two centro-marginal bristles and very numerous short hairs. Post-scutellum dark brown. Wéngs clear, slightly infuscated near the anterior borders, the stronger veins brownish. Venation as shown in the figure (fig. 18a). The costa does not extend as far towards the tip of the wing as in the female of Probezzia pistiae (p. 365); the fork of the fourth vein under the cross-vein. The surface of the wing covered by microtrichia and devoid of longer decumbent hairs; fringe very short on the apical third of the wing. Halteres with pale brown stems and dark brown knobs; knobs bearing a few short hairs. Legs dark brown, conspicuously banded. Femora dark brown, especially those of the Fic. 18. Bezzia foyi, spn. a—venation of wing of female (x 50 cirea.); b—spermatheca (x 210 circa.) hind legs, slightly paler basally, and in the case of the fore and middle legs with a narrow pale band near the apex; knees dark brown; tibiae dark brown, with a narrow pale band near the base, and a less distinct, pale, sub-apical band; tarsi paler, last two segments infuscated. Femora not swollen; fore femora armed with two short, dark, ventral spines on its apical third, middle and hind femora unarmed. Tibiae unarmed, not swollen. First tarsal segment about twice as long as the second on the fore legs, relatively longer on the middle and hind legs; bulbous hairs conspicuous on the hind tarsus, forming two rows on the first and second segments, and one on the third, on the middle legs are single rows of similar hairs on the first and second segments. Fourth tarsal segment cordiform; fifth unarmed. Claws simple, equal, short, less than one-half the length of the fifth tarsal segment. Empodium 363 rudimentary. Adédomen dark brown, venter paler than the dorsum, scantily clothed with short dark brown hairs. Spermathecae two, highly chitinised, unequal, oval or egg-shaped and _ slightly constricted sub-apically (fig. 18 4); lengths about 103@ and 84m in the single female examined, and greatest breadths 53“ and 49 respectively ; the duct narrow (about 4m), and chitinised for some distance (about 25). NIGERIA (Southern provinces) : Lagos, July, 1921 (Dr. H. Andrew Foy); a single female taken in the evening upon the white lining of a lamp-shade. We have pleasure in dedicating this species to the collector, Dr. H. Andrew Foy, to whom we are also indebted for numerous other specimens of Ceratopogoninae from Lagos. Genus PROBEZZIA, Kieff. The chief generic characters of Prodezzia, according to Kieffer (1919), are—eyes glabrous, last four antennal segments in the male elongated, wings bare or covered with microscopic setae, first and third veins separated for their entire length, bifurcation of the fourth vein scarcely proximal to or under the cross-vein, femora unarmed, fourth tarsal segment cordiform, fifth unarmed in both sexes, and claws small, about one-third the length of the fifth tarsal segment, equal and simple in both sexes, or with a small median tooth in the female. The two species which we have assigned to this genus differ slightly in two respects from the above description, namely, in having only the last three antennal segments definitely elongated, the twelfth segment being but slightly longer than the eleventh, and the fcurth tarsal segment short and broad but not definitely cordiform. These differences are very slight, or may even depend on the manner of interpretation of the terms elongated and cordiform. . Both the species described here were reared from plants of the water lettuce Pistia stratiotes, but we were successful in procuring for examination the early stages of the first species only. The larvae are slender, almost white, eel-like organisms similar to those of Culicoides, but larger and with relatively longer and narrower heads. They resemble the figure of the larva of Palpomyia longipennis 364 given by Malloch (1915). They appear normally to inhabit the basal portions of the roots of the Pzs¢za plants, and were reared to the adult stage from plants taken from the water and brought, or sent through the post, to the laboratory, and subsequently kept merely moist. They are, however, capable of leading an aquatic existence, and move very rapidly in water, swimming about in a manner similar to the larvae of Culicoides. At the posterior end of the body are long, stout hairs, which are an aid'to progression. The pupae are similar in form to those of Cudicotdes. They are able to survive in water, but when placed in it quickly make for the side and wriggle themselves above the surface. In the latter situation they remain practically sedentary if undisturbed. The duration of the larval stage was not determined, that of the pupal stage was two to four days. Probezzia pistiae, sp. n. MEASUREMENTS. Male. Female. Length of body... eee ae nes won, J-O mmm. 2°5 mm. Length of wing AM soe oar 20 .. 2 Temi. I'9 mm. Greatest breadth of wing ... ou 39 .. O4mm. o6 mm. The male is a much smaller, and much darker brown insect, than the female. Mead: occiput dark brown, with brown hairs. Eyes glabrous, separated in both sexes. Clypeus and proboscis brown, with brown hairs. Palpi dark brown, rather slender; the first segment short, segments two, three and five sub-equal, about twice as long as broad, the fourth segment rather shorter; third segment not inflated, without a sensory cup but with a few sensory hairs situated distally on its inner side. Mouth-parts somewhat similar to those of midges of the Genus Pvionognathus. Wabium soft and hairy. Labrum rather strongly chitinised, the proximal two-thirds broad, the distal third tapering, fringed with delicate hair-like processes. Hypo-pharynx broad, tapering distally to a rounded apex, and fringed. Mandibles (fig. 21 d), in the female, similar to those of P. marmoratus, and similarly situated, but without teeth and with only a few delicate hair-like processes on the outer edge; the teeth on the inner edge are seven, large and strong, and proximal to them is a row of delicate hair-like processes; in the male, mandibles smaller, less highly chitinised and without strong teeth, but with about five delicate hair-like processes on the inner side. 365 Maxillae rudimentary: Antennae: dark brown, the last five segments in the female, and the last three in the male, darker than the rest. First segment small, bearing a few short hairs in the female. Torus yellowish-brown in the female, dark brown in the male; bearing a few short hairs. Flagellum segments sub- cylindrical: in the female, segments four to ten from twice to nearly two and a half times as long as broad, segments eleven to fifteen elongate, from nearly five to seven times as long as broad, the fifteenth segment being the longest and ending bluntly ; in the male, the twelfth segment about two and a half times as long as broad, the last three segments elongate, from three to five times as long as Fic. 19. Probexzia pistiae, sp.n., wing of female. (x 50 circa.) Fic. 20. a—Probezzia pistiae, sp.n., wing of male. (x 50 circa); b—Probexzia stepbensi, sp.n., head of male. (x 60 circa.) broad, the fifteenth segment being the longest and ending bluntly. Hairs short and scanty in the female; plume of the male poorly developed. Thorax: dorsum uniformly dark or darkish brown, clothed with short dark brown hairs and bearing laterally and posteriorly a few dark brown bristles. Pleurae darkish brown. Scutellum darkish brown, bearing a transverse row of six to eight bristles, and in the female numerous scattered short hairs; in the male are usually only six bristles, and the hairs are more scanty. Post-scutellum dark brown, without a pit. Wzexgs unspotted. Venation as shown in the figures (figs. 19 and 20a); costa in the 366 female extending further towards the tip of the wing than in the male. Surface of the wing closely covered with microtrichia; no macrotrichia present. Halteres pale brown, with dark brown knobs. Legs almost uniformly infuscated in the male; in the female, more or less banded, femora with a dark band before the apex, tibiae with a dark apical band and a dark band a little beyond the base, distal ends of first four tarsal segments and whole of fifth infuscated. Femora not inflated and without strong spines. Tibiae moderately hairy ; fore and middle legs with a dark apical spine. Tarsus with first segment more than twice as long as the second, fourth short and broad, almost (especially in the male) but not definitely cordiform, fifth unarmed. In the female, the first two tarsal segments of the hind legs with a double row of ‘bulbous’ hairs, third segment and first, second, and third segments of the middle legs with only a single row. Claws equal, short, not more than half the length of the fifth tarsal segment; in the female, each with a small but well developed basal tooth, in the male, smaller, bifid at the tip. Empodium small, hair-like. Addomen brown, clothed with short dark brown hairs, which are most numerous on the posterior segments ; in the female, eighth sternite bearing two highly chitinised plates, one on each side of the vulva, clothed distally with numerous short hairs. Spermathecae two, moderately chitinised, sub-spherical, unequal, in one instance measuring 57“ by 50m and 46p by 4oz, respectively; commencement of duct chitinised for only a short distance (about 4m to 7). Hypopycium (fig. 21 a-c). Hypopygium highly chitinised and d Fic. 21. Probexzia pistiae, sp.n. ato c—male hypopygium. a—ninth tergite ; b—harpes ; c—forceps and aedoeagus. d—mandible of female. 367 dark brown, rotated so that the forceps lie dorsally.* Ninth segment : tergite short, bearing at its posterior end two large, hairy, lateral lobes, between which the spiculated lining membrane of the tergite projects as a blunt process; sternite deep, slightly excavated in the middle. Forceps: side-pieces short and broad, about as broad as long, tapering slightly distally; claspers reduced to a small knob bearing a few long hairs. Harfes highly chitinised, fused in the middle line to form a stout chitinised rod, with a rounded, somewhat expanded distal extremity. Aedoeagus highly chitinised, tapering gradually to a narrow distal extremity; membrane joining the ventral wall of the aedoeagus to the ninth sternite studded with spicules. Pupa. Length, female, about 3 mm., male, considerably smaller, about 2°77mm. Well chitinised; male very much darker than female. Respiratory trumpets usually bent posteriorly, rather short and broad, length about o'2 mm., ratio of length to breadth about 7 to 1, smooth and without knob-like processes, the distal end infuscated. The main tracheal trunk is broad, devoid of lateral branches, and terminates in a number (about eighteen) of short, blunt processes which lead to the surface and are arranged in the form of an inverted U. Cefhalo-thorax of the usual form, but the separation of cephalic and thoracic portions is ill-defined. Anterior marginal tubercle very small, bearing a long bristle; above the highest part of the antennal case (the beginning of the flagellum part) are two socket-liké marks, apparently unarmed; a little further back, and in front of the base of the trumpet, is a small tubercle bearing a minute spine; anterior dorso-median tubercle very small, bearing two small sockets, the inner unarmed, the outer bearing a short, spine-like hair, dorso-lateral situated very close to the base of the trumpet, rounded and irregular in form, bearing a long and a shorter hair and an apparently unarmed socket; ventro-lateral ill-defined, bearing a short hair and an unarmed socket; ventro- median represented by two moderately long hairs. Dorsal tubercles much reduced : anterior double, each half bearing a moderately long hair, lateral bearing a similar hair, posterior bearing a rather smaller one. Anterior to the dorsal tubercles is a transverse row of * It will be convenient, however, for descriptive purposes to continue to refer to the surface on which lies the tergite as the dorsal, and to that on which lie the sternite and the aedoeagus as the ventral. 368 highly chitinised rugae, and posterior and external to the lateral tubercle is an unarmed socket. Postero-dorsal tubercle obsolete. Posterior margin of dorsum rounded, not extended backwards as a median process. Addomen: first segment small and narrow, second large and broad, the others decreasing progressively in breadth towards the apex. Integument spiculated, especially in the male, and with pigmented areas similar to those of Séélobezzia spirogyrae. Anal segment with acutely pointed, slightly divergent, dark-tipped processes. Dorsal tubercles: antero-submarginal, small, situated close together and almost contiguous, each bearing a hair, the outermost the larger ; postero-marginal, four, the inner small, bearing a minute hair, the next merely an apparently unarmed socket, the outer two larger, contiguous, each bearing a hair. Ventro-lateral tubercles: all arising from a central projection of the segment; antero-submarginal, small, bearing a hair, and a little dorsal and shghtly posterior to this tubercle an apparently unarmed socket; postero-marginal, two, well developed, the ventral bearing a hair, the dorsal double and bearing two hairs. _ Ventral tubercles: postero-marginal, three, almost contiguous, the inner bearing a short hair, the other two longer hairs. Larva. The larva is eel-like and slender, pale coloured or nearly white; length 7 mm. to 9 mm., greatest breadth about 0-2 mm. to 03 mm. Head yellowish-brown, long and narrow, length about o'4 mm., greatest breadth about 011 mm. Eyes black, bilobed or reniform, situated laterally a little anteridr to the middle of the head. Antennae and palpi small. Hairs quite small, and some apparently unarmed tubercles also present: on the dorsal surface, one pair anterior admedian situated about the level of -the mandibles, two pairs (the more anterior with a divided hair) and a pair of small, apparently unarmed, tubercles anterior dorso- lateral situated at about the level of the comb-like part of the hypopharyngeal sclerite, one pair of apparently unarmed tubercles central dorso-lateral, two pairs posterior dorso-lateral, and two pairs bearing small spines posterior admedian at the extreme posterior end of the head ; on each lateral surface, one anterior, two about the level of the eye, and a small tubercle, apparently unarmed, a little more posteriorly ; on the ventral surface, two pairs anterior admedian, and one pair central ventro-lateral. Mental plate with a strong, pointed, 369 central tooth, and two more delicate teeth on each side. Hypo- pharynx not very strongly chitinised, the posterior sclerite comb-like, bearing about a dozen pointed teeth. Mandibles large and highly chitinised, base expanded, distal portion a powerful hook. Body cylindrical, composed of twelve elongated segments each bearing a few minute hairs. On the distal end of the anal segment are fourteen stronger hairs arranged as follows: dorsally and ventrally two pairs of long, stout, dark hairs about half the length of the anal segment, and two shorter hairs; laterally, on each side a single short hair; these hairs are usually turned anteriorly, and in life appear to be of assistance in progression. Anal gills of the usual form, rather short, being about one quarter the length of the anal segment, deeply cleft into two pointed processes. GOLD Coast: Oblogo, near Accra, December, 1920, to June, 1921, numerous specimens reared from plants of the water lettuce (Pistia stratiotes), taken from a swamp and from the river Densu (Pl. XXI, fig. 1). The larvae, while quite capable of leading an aquatic existence, appeared normally to frequent the roots of this plant, and were frequently reared to the adult stage in plants kept merely moist. Probezzia stephensi, sp. n. This insect, of which at present we possess only a single male, resembles Probezzia pistiae, and, indeed, was originally included among some examples of that species in our collections on account of its almost identical colour markings. Subsequently certain morphological differences were observed, particularly in the hypopygium, which warrant its separation as a distinct species. Only the chief differences between this species and P. fistiae are here given. MEasurEMENTS. Length of body (one male) Au vod be a: .. I'7mm. Length of wing... are “ee ad ae S .. I'Omm. Soreapent DreadtnGh WIP cc ssc igs een) see a: "4 IML, Head (fig. 20b) facets of eyes very widely separated (by about 105) dorsally; anterior median angle of the occiput broad and obtuse. Thorax with rather fewer hairs on the dorsum, and some 379 of them larger than those in P. #zstzae; scutellum bearing two sub- median and two lateral bristles, and a few (about a dozen) short hairs. Legs: hind legs with a single row of ‘ bulbous’ hairs on the first tarsal segments only. Claws with terminal fork deeper. Abdomen bearing shorter and more scanty hairs. Hyporycium (fig. 22). Nznth segment: sternite reduced to a narrow strip of chitin; tergite short, tapering, bearing only two large bristles which are situated one on each side near the posterior margin, continued posteriorly as two large processes, bearing hairs and a few bristles, which are separated from each other in the middle, and with a large, hairy, lobe-like process on its under surface. Forceps: Fic. 22. Probezzia stephensi, sp.n., outlines of male hypopygium. a—ventral view ; 5—lateral view. (% 250 circa.) side-pieces of usual form, moderately hairy; claspers of usual form, rather short (about half the length of the side-pieces), broad and without large hairs at the base, narrowing abruptly in the middle, and terminating in a somewhat spoon-like end. Harfes in lateral view bent in the middle at a right angle (see fig. 22 6), the distal halves somewhat beak-shaped; in dorso-ventral view the distal halves appear as two contiguous chitinous plates (see fig. 22a). Aedoeagus broad, tapering, bearing on each side at the distal end a barb-like process; ventral wall chitinised almost to the base; membrane joining the aedoeagus to the ninth sternite devoid of spicules. 371 GOLD CoAsT: Oblogo, near Accra, March, 1921; reared from some plants of the water lettuce (Pzstza stratiotes), taken from the river Densu. (Pl. XXI, fig. 1). We have much pleasure in dedicating this species to Dr. J. R. C. Stephens, to whom we are indebted for numerous interesting collections of biting midges from the Ilorin Province, Nigeria. Genus DICROBEZZIA, Kieff. The chief characters of this genus, which otherwise resembles Bezzia, according to Kieffer (1919), are—wings with the third vein reaching as near to the wing apex as the anterior branch of the fourth vein, femora unarmed, fourth tarsal segment cordiform, fifth tarsal segment in the female armed with several pairs of black ‘batonnets,’ in the male unarmed, and the claws in the female equal, bifid, the large branch two-thirds the length of the fifth tarsal segment, the small branch two-thirds the length of the large branch, in the male small and simple. We have referred the following species to this genus largely on account of the fact that the fifth tarsal segment in the female is armed with short, dark spines, but it should be pointed out that in other respects it does not entirely agree with the generic description given above. The differences are considerable, and would, perhaps, warrant the erection of a new genus. The species closely resembles Probezzia pistiae, both as regards the general morphology of the adults, including the structure of the hypopygium of the male, and the characters of the pupa. In the following description, therefore, only the chief points of difference are noted. Dicrobezzia nigritibialis, sp. n. This insect, of which at present we possess only a single male and a single female, resembles closely Probezzia pistiae; the chief differences between it and the latter species are as follows :— MEASUREMENTS. Male. Female Length of body... “ie aad on _cclie Se yrrG 2°7 mm. Length of wing I°7 mm. 2°2 mm. Greatest breadth of wing w me te -. O74 mm. o6 mm. 372 The female is darker than the male. Head very dark brown. Eyes separated in both sexes. Palpi in the male small, stumpy, tapering distally, all the segments very short; in the female, longer, as in P. pistiae. Antennae: torus almost black in both sexes: middle segments of the flagellum paler brown in the male, in the female, antenna entirely dark brown. Thorax: dorsum uniformly dark brown, almost black; hairs scanty. Pleurae very dark brown. Scutellum very dark brown, bearing in the male, two sub-median and two lateral bristles and a few short hairs; in the female four sub- median and two lateral bristles and numerous short hairs. Wzmgs very delicate and with a white appearance, venation as in P. Pisézae ; micro- trichia extremely small and delicate. Halteres with pale yellowish- brown knobs, and darker stems. Legs: femora and tibiae very dark brown, first four tarsal segments pale, slightly infuscated at. their apices, last tarsal segment entirely dark. Fourth tarsal segment in the male longer than in P. #zstiae, cylindrical; in the female, shorter, especially on the fore legs. Claws of the male small, equal, bifid at the tips; those of the female large, each with a well developed basal tooth. Fifth tarsal segment in the female armed with numerous (twelve on the fore legs, fourteen on the middle and hind legs) strong, black spines; in the male unarmed. Addomen dark brown, but not so dark as the thorax, and in the female paler proximally; in the female, eighth sternite’ bearing a few long, stout hairs on each side of the vulva. Spermathecae two, highly chitinised, sub-spherical and unequal (diameters about 76% and 604, respectively); commencement of the duct chitinised for a short distance (about 104). Hyrorycium (fig. 23). Hypopygium very dark and_ highly chitinised. Nznth segment: sternite deep, excavated in the middle posteriorly; tergite not very highly chitinised, bearing at its posterior end two prominent, hairy, lobe-like processes, spiculated portion of its lining membrane prominent. Forceps: side-pieces moderately developed, hairs not very long, distal extremity conical ; claspers obsolete. Harfes very densely chitinised, fused in the middle line, and projecting backwards as a process which does not expand at its end, but appears to be double; in dorso-ventral view the posterior projection appears to be straight but not all in the same focus, in lateral view it is seen to be bent sharply towards the 373 tergite near its distal end. Aedoeagus: form somewhat similar, to that of P. stephensz, but more densely chitinised. Membrane joining the aedoeagus to the ninth sternite studded with spicules. Pupa. The pupa is very highly chitinised, especially that of the female, and coarsely spiculated. Length, male 36 mm., female 43mm. It differs chiefly from the pupa of P. pistiae in the following points. Respiratory trumpets not turned backwards, relatively shorter and broader than those of P. fvstiae, terminal branches of the tracheal trunk only about ten in number. Cefphalo- thorax: dorso-lateral tubercle bearing, apparently, only a single ? b Fic.23. Dicrobezzia nigritibialis, sp.n., outlinesof male hypopygium. a—ventral view of forceps and aedoeagus; 4—lateral view, middle focus showing harpes and aedoeagus. (X 190 circa). hair, ventro-lateral composed of two small nipple-like processes, each armed with a stout hair; ventro-median apparently represented by a very minute hair. Dorsal tubercles very small: anterior double, the one part anterior to the other, each bearing a minute spine; lateral bearing a long hair; posterior bearing a minute spine. Abdomen: cuticle coarsely shagreened ; large, dark, lateral macules at the anterior margin of the segments dorsally, in addition to the macules noted in P. fistiae. Anal segment: cases for forceps small and dark coloured. Dorsal tubercles: antero-submarginal not contiguous, a small hump with a socket-like mark posterior to the outer one; postero-marginal, three, the innermost small, bearing a small spine, the other two situated much more laterally, the inner rather large, bearing a short spine and having on its inner side an 374 _unarmed socket, the outer small, bearmg a hair. Ventro-lateral tubercles: antero-submarginal two, the dorsal one bearing a small spine, the ventral a hair; postero-marginal, two, well developed, each bearing a small spine. Ventral tubercles, three, practically contiguous, the inner bearing a short spine, the middle a long hair, and the outer a short hair. GOLD Coast: Weshiang, near Accra, June 29th, 1921; two pupae found in a sample of the algae growing in one of the reservoirs of the Accra waterworks. We are indebted to Mr. R. Simmons for bringing these specimens to the laboratory. REFERENCES Carter, H. F., Incram, A., and Macrig, J. W. S. (1920 and 1921). Observations on the Ceratopogonine Midges of the Gold Coast. Parts I to IV. Annals Trop. Med. & Parasit., Vol. IV, pp. 187-210, 211-274, 309-331; Vol. V, pp. 177-212. Kierrer, J. J. (1913). Voyage de Ch. Alluaud et R. Jeannel en Afrique Orientale (1911-1912). Insectes Diptéres I. Chironomidae et Cecidomyidae. Paris. (1917). Chironomides d’Amérique conservés au Musée National Hongrois de Budapest. Ann. Mus. Nat. Hung., Vol. XV, pp. 292-364. (1918). Chironomides d’Afrique et d’Asie conservés au Musée National Hongrois de Budapest. Ann. Mus. Nat. Hung., Vol. XVI, pp. 31-136. (1919). Observations sur les Chironomides (Dipt.) décrits par J. R. Malloch. Bull. Soc. Ent. France, No. 10, pp. 191-194. (1919). Chironomides d’Europe conservés au Musée National Hongrois de Budapest. Ann. Mus. Nat. Hung., Vol. XVII, pp. 1-160. Mattocn, J. R. (1915). The Chironomidae or Midges of Illinois with particular reference to the species occurring in the Illinois River. Bull. Ill. State Lab. Nat. Hist., Vol. X, Article VI, May. gee 9.6 Lenin's Ls ae. anol taahy AL. . —_ ee Ripe < ae any . WUSRAGBE aed) Pt Shi aes lest F Mar “gay ollot pas be ifoT z DD yeinew Daher) 3 nd ae sam ce real ona molsivweo\ fi a Siri. » os hn ; ~ aah iti : aeNe Z 376 EXPLANATION OF PLATE XXI. Fig. 1. The river Densu at Oblogo, near Accra, showing the Pistia plants from which were reared the following midges :—Culicoides distenctipennis, Dasyhelea incon- spicuosa, Prionognathus pseudomaculipennis, Kempia ochrosoma, Probezzia pistiae, and P. stephenst. Fig. 2. Pool at Accra, from the marginal mud of which were reared the following midges:—Culicoides austeni, C. distinctipennis, C. neavei, C. schultzet, C. similis, Dasyhelea fuscipleuris, D. inconspicuosa, and Stilo- bezzia spirogyrae. ol. XV ISAT XEXT. Annals Trop. Med. & Parasitol., Vol. XP P Fic. 2 377 THE EFFECT OF SALINE SOLUTIONS AND SEA-WATER ON STEGOMYIA FASCIATA BY Jj. W. S. MACFIE (Received for publication 24 August, 1921) Stegomyia fasctata is relatively intolerant of salt (NaCl). In some experiments carried out in 1915 it was found that the larvae died rapidly in 2 per cent. salt solution, that the gravid adult females were relucant to lay their eggs on this medium, and that if, faute de mieux, they did so the eggs were killed and did not even harden and darken. Moreover, it was found that normal eggs of S. fasciata placed in salt solution of about the same strength (2°3 per cent.) failed to produce living larvae. From these and some previous observations (1914) it was thought that sea-water might be found to be of service in the campaign against this mosquito. More recently these experiments have been repeated at Accra actually using sea-water. It was found that undiluted sea-water killed the larvae in a few hours (two to four), and that when diluted with tap-water 50 per cent. (equal to about 1°6 per cent. NaCl) or over was fatal within twenty-four hours. As regards the influence on the gravid females and the action on the eggs, the results were similar to those obtained with salt solution as is shown by the two following experiments. Experiment I. 20th May, 1919. Stegomyta fasciata, two females and one male in a jar containing sea-water. Females fed this day. 22nd May. Male dead. 23rd ,, Noeggs. Females fed at 10 a.m. 2nth® §5, 4 p.m.—many eggs on the water, all white. sth June. No larvae have hatched. Experiment II, 20th May, 1919. Stegomyia fasciata, one male and one female in a large jar containing sea-water in which stands a small beaker containing tap-water. 23rd May. Female fed. 24th ,, Male dead. 27th ,, Many eggs on the tap-water, a few only on the sea-water: the latter are white. Ist June. Many larvae in the beaker containing tap-water, none in the sea-water, 378 Sea-water, in fact, was found to act in a similar manner to a solution of common salt of equivalent strength. An attempt was made to determine the highest percentage of salt that the larvae could tolerate. Such experiments are not easily devised because normally the larvae pupate after a few days, and it is therefore only possible to determine the percentages which prove fatal rapidly. Two series of experiments were carried out with the object of obtaining information on this point. In the one series larvae were used which were in the state of arrested development, to which reference has been made elsewhere (1915), and showed no inclination to pupate. Such larvae were placed in jars containing 100 ¢.c. or 50 c.c. of water to which each day o71 gm. of salt was added in a 10 per cent. solution; at the end of each experiment the amount of salt present in the medium was determined by titration. In four such experiments in which the strength of the saline medium was increased by o'1 per cent. daily, the larvae did not survive more than 0°8, 09, 0°77, and 0°87 per cent. NaCl respectively. In four other experiments in which the strength of the saline medium was increased by o'2 per cent. daily, they did not survive beyond 1:6, 1°8, o'9, and 171 per cent. NaCl respectively. The results of these experiments were not quite satisfactory because in the control jars containing only tap-water some of the larvae died, showing that either the larvae in this state were delicate or that they were suffering from the lack of suitable or sufficient food. It was thought, therefore, that a better practical test of the amount of salt tolerable to the larvae of S. fasctata would be obtained by starting a culture of these insects in a medium containing a low percentage of salt, allowing it to concentrate naturally by evaporation, and noting the point at which the larvae died. Such experiments it was thought would also show if the larvae were able to become habituated to high degrees of salinity. An experiment was, therefore, started with a natural medium, rich in organic material, in a large jar on the sides of which were very many ripe eggs of S. fasctata. Sufficient common salt had been added to the medium to bring the percentage of NaCl up to 11. The jar, covered only by a piece of gauze, was then placed on the laboratory bench and allowed to concentrate gradually by natural evaporation. The larvae which hatched from the eggs developed 379 rapidly at first, then more slowly, and after about a fortnight appeared to be dying off. On the nineteenth day, when it was clear that they were rapidly diminishing in number, a small quantity of the medium was withdrawn for analysis. It was found to contain 1°3 per cent. NaCl. From this time onwards the larvae steadily dwindled, the last individual dying on the thirty-sixth day of the experiment, when the salinity of the medium was found to be 1'45 per cent. NaCl. During the experiment only a very few of the larvae pupated, and all that did so, excepting the first (which pupated on the ninth day, when the salinity was estimated to be 1'2 per cent.), died in the act. A second experiment on the same lines may be summarised as follows :— 1st Day. Salinity of medium equals 1°1 per cent. NaCl. Multitudes of little larvae which have just hatched. 8th ,, Larvae fewer and not growing much. 16th ,, Very few surviving larvae. No pupae yet. 18th ,, Only two surviving larvae. zoth ,, Last larva dead. Salinity of the medium found to be 1°38 per cent. NaCl. No pupae have appeared. The four experiments of this sort (see table) that were carried out showed that the percentages of salt in the media at the times of the deaths of the last larvae were 1°45, 1°38, 1°45, and 1°45, Percentage of NaCl Number of days for which the experiment lasted respectively. Percentage of NaCl in the medium at the beginning of the experiment in the medium at the end of the experiment, namely, when the last larva died Number of pupae 1, and a few which died. None. None. The larvae had, of course, begun to die off some considerable time before this concentration was reached. Pupation was very seldom attempted, and was usually fatal. The experi- ments furnished no evidence that the larvae could be habituated to such degrees of salinity. 380 The inference from these experiments would appear to be that a lo to 1°4 per cent. solution of common salt, or an equivalent strength of sea-water, would effectually prevent the larvae of this mosquito from developing to the adult stage. It would seem probable that sea-water, if used for such purposes as flushing drains and gutters, scowering market-places, etc., would kill both the larvae and the eggs of S. fasciata, and that even if puddles were left the adult females would be reluctant to deposit their eggs on them, but that if they did so the eggs would be killed immediately. REFERENCES Macriz, J. W. S. (1914 and 1915). Bull. Ent. Res., IV, p. 339, and VI, p. 225. 381 THE PREVALENCE AND CHARACTER OF TUBERCULOSIS IN HONGKONG BY HENRY HAROLD SCOTT, M.D, ,.M-R.C.PaLond:, F.RSik; D. Pd. GOVERNMENT BACTERIOLOGIST, HONGKONG; LECTURER ON SPECIAL PATHOLOGY, HONGKONG UNIVERSITY (Received for publication 12 October, 1921) III. THE MORBID ANATOMY AS MET WITH IN CASES AMONG CHILDREN It is a well-known fact that the primary portal of entry of the bacilli in cases of tuberculosis and the mode of spread of the disease are by no means always easy to determine: in some instances, indeed, one can hardly do more than hazard a conjecture. When we remember that the bacilli may pass through a mucous membrane and even through the walls of vessels and circulate as foreign bodies without setting up any immediate injury, but only more remotely causing changes at some distant site where they finally settle, we must always be cautious against employing too freely the anatomical distribution of the lesions as found in the post-mortem room for the interpretation of their genetic relations. In some instances, again, none of the ordinarily described routes seem to explain the method of spread, as was indicated in some of those mentioned in a previous paper. The extent and distribution of tuberculous lesions in an animal inoculated experimentally depend upon several factors, namely, the number and virulence of the bacilli, the resistance set up against infection by the inoculated animal, the seat of inoculation, and the time which has elapsed since infection. It was found as a result of several experiments in which the same dose of bacilli from the same source was inoculated at the same site (namely, subcutaneously into the left hind leg) into animals of the same species and as nearly as possible of the same weight, that in ten days the adjacent gland was involved; in ten to twenty days the left superficial and deep 382 inguinal and the sacro-lumbar glands, and also, perhaps, the spleen and the retro-hepatic glands. In another ten to fifteen days the liver, the lungs, the bronchial, suprascapular, and cervical glands on both sides showed involvement (Delépine). Since in my series the respiratory portal of entry was that most frequently encountered, these cases will first be dealt with. Albrecht, Ghon, and others hold that there is a special form of tuberculosis in children, consisting of a primary lung focus and resulting from the entrance of the bacilli by inhalation. This focus, they stated, was usually the size of a pea, but might be quite small and was rarely larger than a cherry. In nearly three-fourths of the cases the focus was single. The focus is believed to arise ‘in aggregations of lymphoid tissue in the neighbourhood of small bronchi.’ Around this focus small tubercles are seen, and perhaps reactionary fibrous tissue. In the case of larger foci, hard, dry caseation is usual, and occasionally there is actual cavitation. As regards the situation of the focus, the order of frequency was: right upper, left upper, right lower, left lower, right middle, the first- named site being four times as common as the last. An examination of the mediastinal glands showing involvement will, in many cases, enable one to predict the situation of the lung focus. The lymphatics, deep and superficial, discharge into the broncho-pulmonary glands situated between the branches of the main bronchi and at the hilus; those from the middle and lower portions of the lung into the inferior tracheo-bronchial at the bifurcation of the trachea; those from the upper into the superior tracheo-bronchial in the angle between the trachea and bronchus. There is also a chain of glands each side of the trachea, the para- tracheal glands. Infection across from one side to the other is frequently met with. ‘ The original focus may open into a bronchus, and thus by inhalation a tuberculous broncho-pneumonia is set up; a like result would, of course, follow the perforation of a bronchus by an inherent gland. Again, there may be direct extension from a gland adherent to the pulmonary tissue, while, lastly, by communicating with a blood-vessel, miliary tuberculosis may ensue. It used to be held that gland infection was primary and the lung condition secondary to it, but this would leave unexplained the 383 fact of apparently arbitrary selection of a site remote from the primary gland infection, while the intermediate tissue remained free from disease. Canti examined the bodies of eighty-four children under ten years of age. Of these, there were thirty-three under one year, and sixteen (19°05 per cent.) showed tuberculous lesions. Of these sixteen, ten had foci in the lungs, the largest was the size of a cherry, the average that of a pea. In eight a single focus only was found ; in one case two foci of practically the same age were seen, and one showed several; in this instance, however, one of the foci was cavernous and appeared older than the remainder. In the eight with a single focus, this was found three times in the left lower lobe, twice in the right upper, twice in the left upper, and once in the right middle. In other words, the findings in this series of Canti’s agreed in the main with those of Ghon as affording evidence in favour of the common existence of pulmonary tuberculosis in children. The chief points to which attention is directed are the following, and, in discussing the series of cases considered here, I do not think one can improve upon the lines taken by Canti, and for purposes of comparison it is advisable to deal with the points in this order.: — 1. The almost constant finding of a lung focus when tuberculous mediastinal glands are present, and the close relation of these glands to the lung focus. . The frequent singleness of the lung focus. 3. The constant finding of tuberculous mediastinal glands when a lung focus is present—a corollary of the first. 4. The almost constant absence of a lung focus when the portal of entry appears to be elsewhere. 5. The almost constant absence of evidence that the portal of entry may be elsewhere when a lung focus is present—a corollary of the last. Work at the mortuary here has afforded me _ exceptional opportunities for studying these questions, the number of bodies to be examined is great, the proportion of children very high, and a few weeks’ experience sufficed to drive home the fact that tuberculosis forms a large percentage of the causes of death. The differences between the post-mortem findings in children N 384 dying from tuberculosis and those in adults are considerable. Amongst the first three hundred consecutive cases with which this and the two previous papers are concerned there were two hundred and twenty-five under ten years of age. The remaining seventy-five are insufficient for a study of adult tuberculosis, so this paper will be restricted to dealing with the disease as it occurs in children here. The number (two hundred and twenty-five cases) will provide sufficient basis for argument as to whether the conditions of tuberculosis in the tropics, as exemplified at least in Hongkong, resemble those at home, and, if not, in what the differences consist. For purposes of discussion, it will be well to take the points in the above order. 1. The almest constant finding of a lung focus when tuberculous mediastinal glands are present, and the close relation of these glands to the lung focus. The truth of this statement has been substantiated in the majority of the present series. In twenty-nine instances, however, caseous mediastinal glands were found without any focus being detected in the lungs. Eleven of these showed strong evidence of being primarily alimentary and the mediastinal glands may have become involved secondarily to the mesenteric, a condition the occurrence of which was proved by the experimental work of Calmette, Guérin and Breton (1907). They found that in guinea-pigs dying in two to four weeks after being fed on the bacilli the mesenteric glands (especially the superior deriving from the small intestine) were enlarged and inflamed, although no trace of any intestinal lesion could be determined. After six to seven weeks, these glands were caseous in greater or less degree and the lungs showed involve- ment by miliary tubercles with affection of the corresponding tracheo-bronchial glands. Furthermore, these glands, as shown in several of the cases detailed in this series, become caseous more rapidly than the pulmonary lesions preceding their involvement. In one other of my series there was a tuberculous abscess of the sixth cervical vertebra discharging into the right pleural cavity, which might account for the involvement of the mediastinal glands without a focus in the lung. Putting these aside, there were still seventeen which did not conform to the statement relative to the 385 presence of a lung focus when mediastinal glands are found and the relation of the glands to the focus. Of these seventeen, there were fifteen which showed miliary tubercles in the lungs, occasionally in considerable numbers, but in twelve only a few; nevertheless, mediastinal glands on one or both sides were found enlarged and caseated. In none of them was any sign of tuberculosis found in the tonsils or cervical glands. Where the involvement of the lung with miliary tubercles is fairly extensive, it may be argued that the gland constituted the focus whence the lung became infected, but then we are still in the dark as to the source whence it became itself tuberculous. Apropos of some of these cases, the remarks of Bushnell in the Military Surgeon (1918) may be recalled. He states that from a hilus infection the tubercle bacillus is described as travelling by the peribronchial lymph spaces in a direction opposite to that of the normal lymph flow to a region in the upper. lobes where the lymph motion is most sluggish. This might be aided by a reversal of the lymph current, and such a reversal might in turn occur as the result of a block at the hilus. The tubercle bacilli travelling by these spaces to the parenchyma are resisted by the tissue cells and a type of peribronchial tuberculosis results. Caseation through the bronchus may take place, although peribronchial tuberculosis is more often of the ‘closed’ type, as is evidenced clinically by the frequency with which the bacilli are found in the sputum and the rarity of haemorrhages in such cases. In two others the difficulty is increased by the fact that although the mediastinal glands were enlarged and caseous, in one case the tracheo-bronchial, in the other the paratracheal, forming adherent and caseous masses, there was no involvement of the lungs at all. In the former there were meningeal tubercles mostly at the base and along the sylvian fissures, whereas in the latter the only organs found affected were the kidneys, where at the base of a pyramid towards the lower pole of each were several minute tubercles focally arranged. So much for instances in which caseated mediastinal glands were found without a corresponding focus in the lung. As regards the second point—the close relation of the affected glands to the lung focus—there were four cases in which the two did not 386 correspond, in other words, the expectation of localising the lung focus from the gland involved was falsified. In three the focus was in the left lung, as large as a haricot bean, and in one case there was a cavity as large as a filbert; in the fourth the focus was in the right lung. In each case there was a caseated gland, but on the oppesite side. The related glands on the affected side were, in three, not involved at all, and in the fourth there was a little congestion only. It is well known that communication between the glands from one side to the other may be free, and this may be offered as an explanation of the passage of infection from one side to the other; nevertheless, it is less likely that in children, in whom caseation of the mediastinal glands occurs early and readily, infection should pass to the opposite side and apparently miss those on the side affected. One example may be quoted briefly:—In a boy, three years of age, there was.a sub-apical focus as large as a pea in the right lung and a few scattered miliary tubercles in both; the hilus glands were enlarged on both sides, but whereas on the right they showed merely a small caseous point on section, on the left the gland was completely caseated. This differs from the four previously mentioned in that there was a spot of caseation in the gland on the side of the focus, whereas in the others the related glands had apparently escaped altogether. 2. The frequent singleness of the lung focus. Among the sixteen cases described by Canti there were eight with a single focus, while Ghon states that in 72°35 per cent. this is the case. Of the two hundred and twenty-five children under ten years of age among my series of three hundred cases, there were one hundred and thirty-seven showing focal conditions in the lungs. Of these, there were ninety-five in which the focus was single, 7.é., in 69°34 per cent.; in forty-two, or 30°66 per cent., there was more than one. Ghon found that when only one lung focus was discovered the various lobes were involved in the following order of frequency :— Right upper 30°08 per cent., left upper 23°24 per cent., right lower 22°54 per cent., left lower 15°49 per cent., and right middle 775 per cent. 387 Of the ninety-five cases in my series in which only a single focus was found, the numbers in which each lobe was concerned were :— Right upper twenty-six, or 27°37 per cent. ; right lower twenty-three, or 24°21 per cent. ; left upper eighteen, or 18°95 per cent. ; left lower fifteen, or 15°79 per cent. ; right middle thirteen, or 13°68 per cent. The main differences, it will be seen, are that in his series the left upper and right lower were affected in about an equal number of times, the former slightly preponderating, whereas in mine these were reversed, and, secondly, in mine the proportion in which the right middle lobe was involved was much higher. ‘ As regards those which contained more than one focus, there were thirty-two with two, five with three, and the same number with several. The following was the distribution in cases with two foci :— Both in the upper lobe of the left lung ... Both in the lower lobe of the right lung Both in the lower lobe of the left lung .. Both in the upper lobe of the right cr One each in the lowest and middle lobes of the right ares One each in the upper and lower lobes of the left lung One each in the upper and middle lobes of the right lung One each in the upper and lowest lobes of the right lung One each in the upper lobe of each lung “ wb NN Ea Pp WORN One each in the lower lobe of each lung Of the five instances in which three foci were found, in one case all were in the upper lobe of the right lung; in another all were in the lower lobe of the left. Of the remaining three, all were in the right lung, viz., one in the upper and two in the lower in two cases, and the reverse of this, two in the upper and one in the lower in one, Finally, in the five cases in which there were more than three foci, the following was the distribution :—All in the left lower lobe in one; in another, three foci were present in the upper lobe of the left and one in the upper of the right; in a third, foci were present in all lobes except the right upper; in a fourth, in all except the left upper ; in the fifth, in all except the left lower. ' Taking all the cases in which foci were found in the lungs, 2.e., in one hundred and thirty-seven instances, the right upper lobe 388 was involved forty-two times, the right lower forty-one, the left upper thirty-two, the left lower twenty-eight, and the right middle twenty-two. Thus, whereas Ghon found that the left upper was that most frequently involved after the right upper, in my series these were reversed; the right upper and right lower were involved almost an equal number of times, whether we consider merely cases with a single focus or whether we have regard to all instances in which focal lesions were present. 3. The constant finding of tuberculous mediastinal glands where a lung focus was present. In adult cases this has certainly not been my experience here, but it appears to hold good with nearly all cases in children. In twelve of this series, in spite of focal affection of the lung, and even of considerable advancement of the disease, there was no naked-eye involvement of mediastinal glands. In four of these the glands showed microscopically giant cells and a few bacilli. In three others, owing to the early age at which death occurred (twenty-two, twenty-four and twenty-nine days, respectively), although foci were present and the disease was of considerable extent in the lungs, an explanation of the absence of tuberculous affection of the corresponding mediastinal glands may be tendered by suggesting that death took place before there was time for gland involvement to arise. ‘This hypothesis finds a certain measure of support from two others, each of seven weeks old, in whom there was a lung focus and the corresponding mediastinal gland was congested and slightly swollen, not macroscopically tuberculous, but on sections being made giant cell systems and bacilli were seen. On the other hand, the findings in two others appear to deprive this hypothesis of its value. In one of these, a child of three years, there was typical phthisis— tuberculous broncho-pneumonia with ulceration—and in the other, four years of age, the disease had been in existence long enough to produce caries of three vertebrae in addition to a lung focus and several caseating tubercles ; nevertheless, no corresponding gland involvement was found. 389 4. The almost constant absence of a lung focus when the portal of entry appeared to be elsewhere. This is a more difficult matter on which to pass an opinion out here, where the chances of infection by a double route, respiratory and alimentary, are so great. In a previous paper, when we were discussing cases in which the primary portal was uncertain, several of such were dealt with. To take these as instances for considera- tion as to the verity of the dictum of this section would be to beg the question. The statement would apply rather to places where one sees either respiratory or alimentary cases, or at least cases in which the primary portal is undoubtedly one or the other, not those in which the dual route is not only possible, but, as in many here, highly probable. Apart from these, there were five in which a definite lung focus was present, although the primary portal of entry was probably, in fact, one might say certainly, not via the respiratory tract. In two there was a condition cf tabes mesenterica, the glands in the abdomen being in aggregated caseous masses; in one of these cases, an infant of eight months, there was a focus the size of a pea in the right lung; in the other, a child of twelve months, a focus as large as a marble, also in the right lung. In two others, aged twenty months and four years respectively, there was extensive tuberculous enteritis with numerous ulcers and caseated mesenteric glands, and in the second marked tuberculous peritonitis also; in each of these there was also a lung focus. In the fifth, a girl of four years, there was caries of three vertebrae, and several tuberculous ulcers were present in the intestine; in the lower lobe of the right lung was a focus aS large as a cobnut. It is a difficult matter to discuss this question apart from. the next, namely :— 5. Lhe almost constant absence of evidence that the portal of entry might be elsewhere when a lung focus was present. In the previous points, my findings have been to a great extent in agreement, but in this, if I understand it correctly, my experience is distinctly at variance. Several of the cases reported in this series would find an explanation in a dual route of infection, whereas the statement above 502 would appear to rule out such an occurrence, not merely as a more or less simultaneous infection, but even after an interval. So it would seem to put forward the claim that, given a primary lung focus due to respiratory route infection, there is little, if any, likelihood of the intestine becoming affected unless secondarily to the pulmonary focus. This cannot be ascribed to an increased resistance or immunity owing to the presence of the lung tuberculosis, because it is a well-established fact that intestinal tuberculosis can arise from the swallowing of infected sputum. One explanation which, however, in my opinion, savours rather of evading the difficulty than explaining it, would be to say that whenever a lung focus is present together with definite alimentary tuberculosis, the former was the primary site, and the intestinal, though perhaps more advanced, arose from the swallowing of the infected sputum. We may account for some of the cases in this way: the respiratory route gives rise to the lung focus primarily, the intestine becomes infected secondarily, and the lungs are again involved by miliary tubercles spreading via the lymphatics to the thoracic duct, and so to the pulmonary circulation. A considerable number in this series may be thus explained, and several have been mentioned in the previous paper. In connection, however, with these points—the ‘almost constant absence of a lung focus when the portal of entry appears to be elsewhere,’ and the corollary of this, the ‘almost constant absence of evidence that the portal of entry might be elsewhere when a lung focus was present’—there are twelve which did not appear to conform. There is no need to describe them all; two examples will suffice: —(1) A girl of four years with tuberculous caries ‘of the spine. In the discussion on this case, it was stated: ‘The spinal site was probably the oldest; from the number of ulcers in the intestine, from the fact of the large and the smal! both being involved, and from the mesenteric glands being in large caseous masses, the alimentary tract would appear to have been involved prior to the lung.’ There was a sub-apical focus in the right lung the size of a cobnut, becoming caseous. (ii) A female infant of eight months; the mesenteric glands were in large caseous masses, whereas the mediastinal were not much enlarged and contained merely small caseating points. The tuberculous infection of the lung 39% was limited to the middle lobe of the right, in which there was a distinct focus the size of a pea. It has been stated by MacCallum that in children one may find, instead of the apical lesion so common in adults, a caseous softening of bronchial lymphatic glands and erosion through a bronchus to produce wholesale tuberculosis of a lung, or a large section of it. Several of the cases in this series might be looked upon as examples of this; we must bear in mind, however, that, though this may explain generalised infection of part or even the whole of a lung, we still leave unaccounted for the source whence the gland became involved. In those instances in which we find a focal lesion, the subsequent more generalised condition in that lung, or part of it, may be ascribed to reinfection from the gland, which itself was due to the primary focal lesion. In cases where no focal lesion is found to which the mediastinal gland could be traceable, and especially in cases where both lungs are attacked by miliary or grey tubercles, the spread may have occurred by the blood-stream; if general, by the pulmonary circulation, if localised, either by a branch of this or of a bronchial artery. In one case, in which there was caries of . the right side of the sixth cervical vertebra and pus discharging into the right pleural cavity, into a loculus shut off by pleural adhesions, the glands at the hilus and along the trachea were caseous and may have arisen secondarily to the lesion above, and the widespread miliary affection of the right lung may then have resulted by the method which MacCallum describes. Passing on to the question of abdominal and alimentary tuber- culosis, there is not much to be said in this paper dealing with the aspect of the morbid anatomy. As already stated, cases of isolated primary tuberculosis of the intestines, not uncommon at home, are comparatively rarely met with out here. In only four instances was the disease confined to the abdomen; three of these were infants, aged respectively ten, eighteen and twenty-two months, the fourth was a child of seven years. From the intestines with, or more often without, any local lesion, the bacilli are arrested for a time in the mesenteric glands and thence spread either by way of the lymphatics to the blood and so to the lungs (of this, several instances have occurred in this series, and have already received sufficient mention), or else by the portal blood to the liver. This has been 392 very rare in these cases. In fact, cases in which the liver- was extensively involved have been few. The spleen in nearly all was more affected than the liver, and in two only was the liver infected and not the spleen. When tubercles were found in the liver, in the large majority of instances they were small, miliary to pin-head, and confined to the surface. The rarer forms of intestinal tuberculosis where the disease is localised to the region of the ileo- caecal valve to produce a mass of tuberculous cicatricial tissue constricting the lumen, I have met with once among these cases. The following is worthy of mention while dealing with alimentary forms. The subject was a child of two years; there were tuberculous ulcers in the small intestine, and the mesenteric glands were enlarged and caseated. A tuberculous meningitis was the only other lesion detected ; there were numerous pin-head tubercles at the base and a few on the vertex. The mode of extension in this case is obscure, One. only evades the question by saying that it was probably metastatic by way of the blood-stream, analogous possibly to cerebral abscess in cases of haemorrhoids and in liver conditions. The meninges and brain were found involved in a considerable number of cases, particularly the former, and usually as miliary tuberculosis affecting mostly the base and the Sylvian fissures. Definite focal masses in the brain or cerebellum were comparatively rare. There were several cases in which the lungs and meninges were the only parts in which tubercles were found. In seven instances the meninges and one lung only were affected ; in six of these it was the right lung which was attacked. In one other case, in addition to the lung and meningeal involvement, there was a tuberculous focus the size of a small marble in the right cerebellar hemisphere. In one case, a girl of eight years, the mediastinal glands and the meninges were the only parts found affected, there being no tubercles, either focal or miliary, in the lungs. The mode of extension to produce the peculiarly limited distribution of tubercles in lungs and meninges is obscure, and is, perhaps, analogous to brain affections secondary to other pulmonary conditions, bronchiectasis, for example. There is not, as far as I am aware, any lymphatic connection between the lungs and the base of the brain, and if the extension occurred by way of the 393 blood-stream, why should the secondary infection be so limited ? One would almost certainly expect to find signs in other organs. I am also unable to suggest any significance for the fact that, in the combination of lungs and meninges, of the seven cases in which one lung only was affected, in six it was the right; and of the five cases in which both were involved, in three the right was more affected than the left. Again, in the case already referred to in which the mediastinal glands and meninges were tuberculous, but in which no signs were detected in the lungs, the glands were those of the right side only. The combination of lungs, meninges and spleen was a little more common, fifteen of such being met with. In two of them only one lung was involved, in each case the right. In this connection, it may be worth noting that the involvement of the spleen with miliary tubercles as the only abdominal viscus affected has not been very infrequent, and may find explanation, perhaps, if one regards the spleen as the meeting point of lymphatic and blood terminals. I may refer here to the observations of Dumas upon what he calls an unusual form of tuberculosis met with in Salonica amongst Senegalese and Arab troops. He noted that the mediastinal and peribronchial lymphatic systems were first affected, and later the pleura and pericardium. In the early stages the glands were merely enlarged, but later they suppurated or became caseous. At the onset there were no lesions of the parenchyma of the lung, and even later not the ulcerating and caseous form, only a few scattered tubercles. He considered that it was in the spleen that the lesions passed from the lymph to the blood stream. When the spleen was normal no tubercles were found in the lungs, but when the former was invaded the latter were also affected. Examples of this in children, 7.e., cases in which the spleen is the only abdominal viscus involved and the connection referred to between this and the pulmonary findings seem to find support in some of the instances in this series. One may be given: a male child, three years of age, showed miliary tubercles scattered through both lungs, the spleen contained similar tubercles, as did also the meninges. The hilus and paratracheal glands on the right side were enlarged to the size of a cobnut and were caseous. Again, as was pointed out above, in the peculiar limitation at times to the 394 lungs and meninges, if haematogenous metastasis occurs, the first part to suffer is the meninges. In two other instances, however, the lungs were affected with small tubercles (no focus) and the hilus and paratracheal glands were caseated, but the spleen was not involved ; in one of these blood infection was apparent, since the meninges revealed basal tubercles. On nine occasions were focal tubercles found in the brain or cerebellum. The commonest site was the latter, for cerebellar were found in eight. Of these, the focus was present in four cases in the left hemisphere, in three in the right, and once in both. In this last there were four distinct foci, two in each lateral lobe; two others had a second, extra cerebellar, focus, namely, in the right cerebral hemisphere in one, in the left crus in the other. Finally, one subject, a female child of two and a half years, exhibited multiple foci: three distinct ones in the left lobe of the cerebellum, two in the left cerebral hemisphere, and one in the left hippocampus. A striking feature of the tuberculous conditions as met with here is the rarity of bone, joint or skin affections. In this series there were five with tuberculous ulcers of the skin, four of these on the face and neck and one on the forearm. Not a single instance of tuberculous joint disease was encountered, and only three with bone lesions. In two of these there were caries of the lower dorsal and upper lumber vertebrae; a third had caries of the sixth cervical vertebra, and tuberculous disease also of the left tibia and both femora. A fourth had widespread tuberculous disease—lungs, liver, spleen, peritoneum and meninges—and extensive caries of the left mastoid, but there was no absolute proof that this last was due to tubercle. Tuberculous cervical adenitis, so frequently met with at home, is a comparative rarity in the post-mortem room here. Of the whole series there were only twenty-eight instances, and in three of these the enlargement was very slight, so slight that there was not sufficient macroscopical evidence to determine its nature; on section, however, giant cells and tubercle bacilli were seen. In one case there was almost universal involvement of the glands—cervical, submaxillary, supraclavicular, axillary, media- stinal, mesenteric and femoral—together with widespread pulmonary, intestinal, and some renal tuberculosis. This was a child of only 395 three years. Sections did not show any of the usual conditions characteristic of Hodgkin’s disease. In contrast to this, may be incidentally mentioned a little girl of seven months, showing but one cervical gland enlarged, with very extensive tuberculous disease—lungs, pleura, mediastinal glands, mesentery, intestines, liver, spleen, kidneys, meninges. Cases in which the genito-urinary tract has been involved have been comparatively few in my experience here. The majority of these showed a few miliary tubercles as emboli in the glomerular capillaries, consituting part of a general haemic infection. There were ten instances in which the kidneys were found focally affected in conjunction with widespread tuberculosis. One, a girl of three years of age, merits further mention on account of the peculiar distribution. The extensive alimentary infection—tuber- culous ulceration of both large and small intestines, caseated, adherent mesenteric glands, extensive infection of the peritoneum— and the equal distribution of tubercles throughout both lungs would point to the alimentary canal as the primary portal of entry. The condition of the kidneys, however, does not support the idea of a haematogenous origin from entrance of the bacilli into the general circulation from the lungs. Each kidney showed a large focus in a similar situation at the lower pole, the size of a cobnut and caseous, most advanced at the margin between the cortex and a pyramid, and passing in as if later it would discharge into the pelvis. No indication of involvement of the ureters was detected. The foci gave the impression that they had arisen from affection of the tubules in course of excretion (as mentioned by Aschoff and Israel). The foci were each of them.in a more advanced state than a few smaller, pin-head and miliary, tubercles in the cortical area of the right kidney, which had more likely arisen by haematogenous infection. The kidney foci appeared to be of considerably older standing than the pulmonary condition, and showed more advanced caseation than the mediastinal glands, but less than the mesenteric. There were no indications of an ascending infection from lower down the urinary tract. Other viscera, liver and spleen, showed sparse tubercles only, and these were, in the former at all events, confined to the peritoneal surface. Briefly, the age of the kidney condition appeared to be less than that of the alimentary but 396 more than that of the pulmonary, and the mode of involvement is obscure. Two others are of sufficient interest to warrant brief mention :— G) An infant of nine months, showing generalised tuberculosis— lungs, intestines, liver, spleen, kidney—and, in addition to miliary infection of the last, one calyx in the left was hollowed out and lined by tuberculous material; (41) a girl of four years, with miliary tuberculosis of the lungs and of the liver surface, but the only focus found in the body was a caseated mass occupying practically the whole of a pyramid in the left kidney. Finally, while speaking on the subject of genito-urinary affection, mention must be made of two cases of exceptional interest. Both exhibited extensive disease, lungs, meninges, intestines, peritoneum and ovaries. In one, a girl of four years, both ovaries were enlarged, and had become converted practically into caseated masses; this case has been described in a previous paper when discussing the primary portal of entry. In the other, an infant of only eight months, the Fallopian tubes were swollen and caseous on both sides, while the ovaries were in a condition similar to the last. In this case a possible (or, rather, probable) source was by contiguity from the tuberculous peritoneum, but in the former the tubes did not appear to be affected. REFERENCES Ascuorr and Israzr’(1g1g). Cited by MacCallum. Text-book of Pathology, p. 616. BusuneLt, G. E. (1918). Military Surgeon. Catmette, Guerin, and Breton (1907). Ann. de l'Institut Pasteur, Vol. XXI, No. 6. Cantr, R. G. (1919). Primary Pulmonary Tuberculosis in Children. Quarterly Fourn. Med., Vol. XIII, No. 49, pp. 71-81. Deering, S. Contribution to the Study of Delayed or ‘latent’ Tuberculous Infection. Ann, de V Institut Pasteur. Dumas, A. (1919). Adenopathie trachéo-bronchique 4 marche rapide. Lyon Méd., pp- 180-187. Guon, A. Die primare Lungenherd bei der Tuberkulose der Kinder. MacCuttum, W. G. (1919). Text-book of Pathology, p. 607. 397, ON THE GENUS CYLICOSTOMUM* BY AD. Hic WEEE (From the Zoological Laboratory, Veterinary College, Utrecht) (Received for publication 29 October, 1921) Last year (1920 a, p. 268) I published an enumeration of the species of the genus Cylicostomum, and divided this genus. into eight groups, of which three had been distinguished by Looss (1902, pp. 130-132). Since that time some new species have been described. Moreover, I think it necessary to modify somewhat the groups, so that I now divide the Cylicostomum species as follows :— I. Letracanthum-coronatum group. The external leaf-crown is composed of eighteen to twenty-four elements ; those of the internal leaf-crown are thin, triangular plates, the place of origin of the latter extending in a backward direction to some distance from the anterior margin of the mouth-capsule. Mouth-capsule rather short. Posterior extremity of the 9 straight or slightly bent in a dorsal direction. * Of late, some authors (Railliet, Travassos) use, instead of the name Cylicostomum, the genus-name Trichonema, which ought to be used according to the law of priority. In 1874, * Cobbold (Veterinarian, Vol. XLVII, p. 85) described under the name Trichonema arcuata larvae of Cylicostomum. One year later (Veterinarian, Vol. XLVIILI, p. 241) he established the fact that this Trichonema is the larva of ‘ Strongylus tetracanthus, the present genus Cylieostomum Railliet. As, however, the law of priority must be applied ‘ when any stage in the life-history is named before the adult,’ and the name Cylicostomum (Railliet, L’ Echo V étérin., "Vol. XXXI, p. 40) is used in 1901 for the first time, Trichonema ought to be used. It does not seem advisable to me in this, nor in the like cases, to adhere to the rules of zoological nomenclature, when, owing to this, a completely familiar name would be replaced _ by one still entirely unknown. Neither did I always adhere to the law of priority in my revision of the Trematodes, Cestodes and Nematodes of domestic animals, in the third edition of Sluiter and Swellengrebel, ‘ De dierlijke parasieten van den mensch en yan onze huisdieren.’ 398 . tetracanthum (Looss), 1902, p. 124. . labratum (Looss), 1902, p. 124. H.* . ornatum, Kotlan, 1919, p. 10 . labtatum (Looss), 1902, p. 125. . labiatum (Looss) var. digitatum, n. var. H. . Sagittatum, Kotlan, 1920, p. 4. . coronatum (Looss), 1902, p. 125. H. Bwh a. aN eG Sy SH) Sorry ion Own Il. Alveatum-catinatum group. The external leaf-crown consists of twenty to twenty-nine elements; those of the internal leaf-crown are similar to those of the first group. Posterior extremity of the Q strongly bent in a dorsal direction, with a swelling before the vulva, so that it reminds one of a human foot, seen laterally. 7. C. alveatum (Looss), 1902, p. 127. 8. C. catinatum (Looss), 1902, p. 128. Sane. Psa las (Looss), var. Oe Yorke and Macfie, 1920, p. OnenGe 5 edi Yorke and Macfie, 1919, p. 273.. H. 10. C. pateratum, Yorke and mae Ree Pp. 57 (= C. cyma- tostomum, Kotlan, 1919, p Hi: 11. C. gold, Boulenger, 1917, p. 210. H. 12. C. tridentatum, Yorke and Macfie, 1920, p. 153. 13. C. mettami, Leiper, 1913, p. 460. C. tridentatum and C. goldi are evidently closely allied species, as I found the three teeth of the oesophageal funnel of C. tridentatum, described by Yorke and Macfie, also in C. goldz. They are visible only in thoroughly transparent specimens with clean mouth-capsule. It seems to me that C. pseudocatinatum must be considered a variety of C. catinatum. The former does not differ more from C. catinatum than does C. catinatum var. litoraureum, Yorke and Macfie. Both differ from C. catinatum in the appendages of the genital cone. Moreover, the appendages of C. catinatum litoraureum are more like those of C. pseudocatinatum than like those of the typical ca¢znatum form. So when we consider litoraureum a variety as Yorke and Macfie do, this also applies to pseudocatinatum. * The species occurring in Holland are marked H. 399 Ill. Radiatum-elongatum group. Mouth-capsule with a hoop-like thickening at the posterior margin. Elements of the internal leaf-crown generally small and numerous, originating close to the anterior margin of the mouth- capsule. Posterior extremity of the 2 straight, or slightly bent in a dorsal direction. ¢ 14. C. radiatum (Looss), 1902, p. 129. 15. C. triramosum, Yorke and Macfie, 1920, p. 175. 16. C. elongatum (Looss), 1902, p. 129. H. 16a. C. elongatum (Looss) var. Kodldani, thle, 1920, p. 269 (= C. elongatum var. macrobursatum, Kotlan, 1920, p: 9)jo8. 17. C. imsigne, Boulenger, 1917, p. 207. H. 18. C. zebrae, Boulenger, 19204, p. 102. 19. C. adersi, Boulenger, 1920, p. 30. 20. C. nassatum (Looss), 1902, p. 128. 20a. C. nassatum (Looss) var. parvum, Yorke and Macfie (1918, p. 400). 21. C. leptostomum, Kotlan, 1920, p. 3. H. 22. C. auriculatum (Looss), 1902, p. 130. The @ of the species last mentioned forms a transition to the alveatum-catinatum group by the form of its posterior extremity. In this-group I now also include C. leptostomum, Kotlan, a species about which Kotlan published a description in Hungarian only; the author was so kind, however, as to send me a German translation. C. leptostomum is closely allied to C. nassatum, from which it is distinguished by the posterior extremity of the 9 being straight. IV. Calicatum group. Mouth-capsule mostly cylindrical or trapezium-shaped in optical section. Elements of the internal leaf-crown mostly short and implanted in the immediate neighbourhood of the anterior margin of the mouth-capsule. Posterior end of the Q mostly straight, 23. C. calicatum (Looss), 1902, p. 127. H. 24. C. minutum, Yorke and Macfie, 1918, p. 405 (= C. cali- catum var. minus, Kotlan, 1920, p. 6). H. 400 25. C. longibursatum, Yorke and Macfie, 1918, p. 400 (= C. calicatiforme, Kotlan, 1919 a, p. 559 = C. nanum, Thle, 1919, p. 720). H. 26. C. hybridum, Kotlan, 1920, p. 5. 27. C. poculatum (Looss), 1902, p. 126. H. The last mentioned species differs considerably from C. calicatum (e.g., in the bursa, which is closed all round and has a finely . denticulated border) so that Looss (1902, p. 132) did not group them under one head. V. Euproctus-bicoronatum group. The external leaf-crown mostly consists of numerous slender elements; the elements of the internal leaf-crown are extremely large. The posterior extremity of the 9 is straight. 28. C. euproctus, Boulenger, 1917, p. 204. H. 29. C. bicoronatum (Looss), 1902, p. 125. H. 30. C. chlei, Kotlan, 1921, p. 300. H. 31. C. ultrajectinum, Thle, 1920 a, p. 269; 1921, p. 372. H. For the present, some heterogenous elements are taken together in this group, characterized by the possession of very large elements of the internal leaf-crown. C. ihlei is closely allied to the genus Poteriostomum in structure of mouth-capsule and leaf-crowns. The fact, however, that the bursa. copulatrix of this species shows the typical characteristics of Cylicostomum and differs from that of Potertostomum, supports the opinion, shared by Yorke and Macfie (1920, p. 159), that Poterio- siomum must be considered an independent genus contrary to Kotlan’s view (1921, p. 299). Formerly, I considered C. wultrajectinum to belong to the vadiatum-elongatum group, with which it agrees in the thickened posterior margin of the mouth-capsule, from which it differs, however, in the size of the elements of the internal leaf-crown. It is, however, also considerably different from the other species of the euproctus-bicoronatum group in size and number of the elements of the external leaf-crown. Yorke and Macfie (1920, p. 162) expressed the opinion that C. w/rajectinum might belong to the genus 401 Poteriostomum. It has become evident, however (thle, 1921, p. 372), that the bursa copulatrix resembles that of the other Cylicostomum species in every respect, so that it is certain that this species belongs to the genus Cylzcostomum. VI. Brevicapsulatum group. Mouth-capsule extremely short. Posterior extremity of the 9 straight. 32. C. brevicapsulatum, thle, 1920, p. 562. H. 33. C. prionodes, Kotlan, 1921, p. 305. These two species differ considerably, but they can be grouped together for the present. VII. Montgomeryi group. The dorsal and ventral walls of the mouth-capsule are much longer than the lateral walls. 34. C. montgomeryi, Boulenger, 1920 a, p. 104. I will conclude by describing a new variety of C. labiatum (Looss), of which I found a small number of specimens in the large intestine of the horse in Holland. Cylicostomum labiatum (Looss) var. digitatum, nov. var. The new variety agrees in the main (mouth-collar, mouth- capsule) with the typical form. 36 7 to $8 mm., the immature 2? 8 to 83 mm. long. Oesophagus 345 to 410m long. The bursa copulatrix has a dorsal lobe, which varies in length and is mostly longer than in the typical form. The distance from the extremity of D3* to the point of origin of the postero-external ray varies from 310” to 380”. The posterior part of the dermal collar shows the processes occurring in the typical form. The chief difference from the typical form is that the new variety shows finger-like processes on both sides of the posterior part of the genital cone, which are absent in the typical form. The number and form of these processes perhaps fluctuate. (In one case I counted three on each side.) The posterior extremity of the ? is like that of the typical form. The distance from the vulva to the * The short stem which the dorsal rays have in common, I name D®, the two main trunks D, the side branches, from before backwards, D1, D2, and D8. In Cylicostomum D3 forms the immediate continuation of D, 402 anus is smaller however (ca. 95“) than in the typical form; the distance from the anus to the posterior extremity is 86m to 110p, ant Fic. 1. Cylicostomum labiatum Fic.2. Cylicostomumlabiatum Fie. 3. Cylicostomum labiatum var. digitatum. Dorsal lobe of the var. digitatum. Processes of the var. digitatum. Posterior extremity bursa. genital cone, lateral view. of female, lateral view. measured along the axis of the body. The conical end of the body is bent somewhat in a ventral direction. REFERENCES Bourencer, C. L. (1917). Sclerostome parasites of the horse in England. II: New species of the genus Cylichnostomum. Parasitology. Vol. IX. ——— (1920). Sclerostomes of the donkey in Zanzibar and East Africa. Parasitology, Vol. XII. — (1920a). On some Nematode parasites of the Zebra. Parasitology, Vol. XII. Tue, J. E. W. (1919). Cylicostomum nanum, een nieuwe Strongylide van het paard. Tijd- schrift voor Diergeneeskunde. Dl. 46. ——— (1920). Cylicostomum brevicapsulatum, n.sp., eine neue Strongylide aus dem Darm des Pferdes. Centralblatt f. Bakt. Abt. 1. Orig. Bd. LXXXIV. ——— (1920a). Bemerkungen iiber die Gattungen Cylicostomum, Poteriostomum und Craterostomum. Centralbl. f. Bakt. Abt. 1. Orig. Bd. LXXXY. ——— (1921). Das Mannchen yon Cylicostomum ultrajectinum. Centralbl. f. Bakt, Abt. 1. Orig. Bd. LXXXV. - KotrAn, A. (1919). Beitrag zur Helminthologie Ungarns. I. Neue Sclerostomiden aus dem Pferd. Centralbl. f. Bakt. Abt. 1. Orig. Bd. LXXXIII. — (1920). (Hungarian paper). Allatorvosi Lapok 1920. (1921). Two new Cylicostomum species of the horse, Ann. Trop. Med. © Parasit., Vol. XIV. Lerrer, R. T. (1913). A new Cylicostome worm from the horse in London. Veterin. Fourn., Vol. LXIX. Looss, A. (1902). The Sclerostomidae of horses and donkeys in Egypt. Rec. Egypt. Govern. School of Med., 1901. Yorxr, W., and Macriz, J. W. S. (1918). Strongylidae in horses. I. Cylicostomum longi- bursatum, sp.n.; II. C. minutum, sp.n.; II. C. nassatum Looss var. parvum. Ann Trop. Med. & Parasit., Vol. XI. (1919). Idem. VWI. C. pseudo-catinatum, sp.n. Ibid, Vol. XII. ——— —— (1919a). Idem. VII. C. pateratum, sp.n. Ibid, Vol. XIII. (1920). Idem. IX. Cylicostomum: tridentatum, sp.n.; X. On the genus Poteriostomum (Quiel). XI. Species found in West Africa and Jamaica; XIII. Cylicostomum triramosum, spn. Ibid, Vol. XIV. 493 NOTES ON AUSTRALIAN CESTODES BY .P. A. MAPLESTONE (Received for publication 17 November, 1921) INTRODUCTION Whilst on the staff of the Australian Institute of Tropical Medicine, Townsville, a systematic examination was made of the large collection of Cestodes in the Museum. In addition to a considerable number of new species, the description of which will appear in subsequent papers, a number of previously described worms were found in new hosts. I. PREVIOUSLY DESCRIBED CESTODES IN NEW HOSTS Cittotaenia tachyglossi, Johnston, 1911 = Cittotaenia, sp. nov., Nicoll, 1914. Nicoll (1914) recorded a new species of Cittotaenia from the Echidna, Tachyglossus aculeatus, Shaw, but without furnishing any description of it. Detailed re-examination of the same material proves it to be Czttotaenia tachyglossi, Johnston, but the dimensions are all somewhat greater than Johnston gives in his description, notwithstanding the fact that the worms are in an immature state similar to those from which the original description is given. Diorchis flavescens (Krefft), Johnston, 1912. The original description of this worm was published by Krefft (1873), who named it Taenia flavescens, but H. Johnston (1912)! re-examined Krefft’s material and placed it in the genus Diorchis. Up to the present, this cestode has been found in Anas superciliosa, Gmel., the Black Duck; Spatula rhynchotis, Lath., the Blue-winged Shoveller; Netéion castaneum, Eyton, the Teal; and Aythya 404 australis, Gould, the White-eyed Duck. A few badly preserved fragments of apparently thé same species have been found by the writer in material taken from Dendryocygna arcuata, Cuvier, the Whistling Duck. O phiotaenia longmani, Johnston, 1916. Johnston (1916) originally described this species from the snake Asfidotes ramsayi, which wasitaken at Yarelba, Western Queensland. The same species was recently found by the writer in the intestine of Python spilotes var. variegatus, Gray, the Carpet Snake, which was killed near Townsville, North Queensland. Ophiotaenia hylae, Johnston, 1912. O. hylae was described by Johnston (1912)? with the frog, Hyla aurea, as its host, which was captured near Sydney, New South Wales. A scolex and several proglottides of this species were found in the intestine of D. arcuata, the Whistling Duck, which was shot near Townsville. This is strange, as the family Proteocephalidae, to which the above species belongs, has hitherto been recorded only in Amphibia, Reptilia, and the dog. The most probable explana- tion of this occurrence is, that the duck had swallowed a frog infected with the cestode, and the frog had been sufficiently digested to liberate it into the bird’s intestine, without itself having undergone digestive changes up to the time the duck was shot. Acanthotaenia gallardi, Johnston, 1911. This cestode. was originally described by Johnston (1911)? under the name Pvoteocephalus gallardi, and subsequently placed in the genus Acanthotaenia. On this occasion it was found in the intestine of Pseudechis porphyriacus, Shaw, the Black Snake. Since then it has been recorded by Johnston (1912)? as occurring in Pseudechis australis, Gray, the Northern Black Snake; Notechzs scutatus, Peters, the Tiger Snake; and Denisonia superba, Gunth, the Copper-headed Snake. To this list of hosts the writer is now able to.add Dipsadomorphus fuscus, Gray, the Brown Tree Snake, 405 a specimen of this snake harbouring this worm being killed near Townsville. Moniezia alba, Blanchard, 1891. Johnston has recorded the presence of this cestode in Australia in sheep in New South Wales, and the writer has recently obtained specimens of the same worm from a bullock, slaughtered in Townsville, and which came from near Hughenden, Western Queensland. This is the first record of this cestode in the above host in Australia. Thysanosoma giardi, Stiles, 1893. Johnston has noted the occurrence of this worm in sheep in New South Wales, but no note of its frequency and distribu- tion has been given. Over a period of several months, the writer has had the opportunity of examining many worms taken from sheep in Townsville, and they all proved to be of this species. The sheep came from districts representing a wide area in Western Queensland, so it seems that it is widely distributed, and very common, but as far as could be ascertained from inquiries among pastoralists and butchers, it does not give rise to the serious patho- logical condition among sheep, that it does in other parts of the _world. REFERENCES Jounston, T. Harvey (1911).1 Cestoda and Acanthocephala. Report Aust. Inst. Trop. Med. (1g11).2 A new Cestode from the Black Snake. Annals of the Queensland Museum, No. Io, p. I. : — (1912) Re-examination of the types of Krefft’s species of Cestodes. Rec. dust. Museum, Vol. TX, No. 1. — (1g12).2 A Census of Australian Reptilian Entozoa. Proc. Roy. Soc., Queensland, Vol. XXIII, p. 233. — (1912).3 Notes on some Entozoa. Proc. Roy. Soc., Queensland, Vol. XXIV, p. 63. (1916). Helminthological notes. M. Queensland Museum, Vol. V, p. 186. Krerrt (1873). Trans. Entomological Soc. N.S. Wales, 11, pp. 206-227. Nicott, W. (1914). Remarks on the worm Parasites of Tropical Queensland. Med. Fourn. of Aust., p. 244. \o v ‘ , . : 2 ‘ « : 2 ie |" ad. hi aheeres. NY a*? i ee ve Mea: 1: OFF J wh ie j t D1 Ap ry as So MAA . ii ie a> te , aud ' iat - iy * 407 NOTES ON AUSTRALIAN CESTODES BY P.. A. MAPLESTONE (Received for publication 25 November, 1921) II, ANGULARIA AUSTRALIS, sp. nov. This cestode was found in considerable numbers in the intestine of a Stone Curlew (Burhinus grallarius, Lath.), shot near Townsville, North Queensland. It is probably rather rare, this being the only occasion on which it was encountered. EXTERNAL ANATOMY. The worms are very small; the largest individual measured only about 3 mm. long; from its appearance the strobila was complete, but it had not quite reached full development. The number of proglottides in a chain varies between fourteen and twenty (fig. 1). Fic. 1. A. australis. Appearance of worm asa whole. X 35. On account of its small size, there are no macroscopic characters by which it may be recognised, so the following description of the appearance as a whole had to be determined under the low power of the microscope. i 408 Head. The head bears a relatively long, thin rostellum, which was unfortunately broken off in most-cases. From base to tip the rostellum is about 150” to 200m long, and it is about 14m thick. It arises from a bulbous muscular structure, which lies i a deep fossa. It projects directly forwards as a long proboscis-like organ of uniform diameter, except at the tip where it expands into a globular enlarge- ment, about twice the diameter of the stem (figs. 2 and 3). AMG, det. Fic. 3. A. australis. Tip of rostellum, shewing disposition of hooks. X 225. Fic. 2. A. australis. Head, and Fic. 4. A. australis. Hooks. X 450. anterior parts of strobila. x 70. The hooks, which measure about 25 in length, are arranged in a zigzag line which forms four acute angles anteriorly and four acute angles posteriorly. Between an anterior and a posterior angle there are six or seven hooks. Thus, the total number of hooks is forty-eight or fifty-six. Their disposition and shape are shown in figs. 3 and 4. 409 The scolex is about as broad as it is long (250). The four suckers are relatively large oval structures, with their: long axes antero-posterior; they are not exactly oval, being slightly narrower anteriorly. They measure about 130 in length and 60, in breadth, and their lips are provided with cuticular expansions, which project beyond the surface of the scolex (fig. 2). Segments. For about the anterior half of the strobila the segments are quite narrow and rudimentary, the only change being a slight increase in length. But from about the mid-point of the strobila, the proglottides rapidly increase in size, and the reproductive organs reach complete maturity in the course of three or four segments. There are only three or four mature segments in each strobila, the genitalia undergoing a sudden atrophy when the uterus begins to develop, so that the two or three terminal segments contain, beside the uterus, only remnants of the reproductive organs (fig. 1). The shape of individual segments is somewhat uncommon. Each one is like a truncated cone with the narrow end anterior, and the sides, which are concave in the anterior immature proglottides, become slightly convex in the posterior mature and gravid segments. The posterior surface of each proglottis is oval and slightly depressed in the centre, and into the middle of this depression the narrow anterior margin of the succeeding segment fits; the result of this is that the posterior borders project all round beyond the anterior portion of the following proglottis. There is no neck, but posteriorly the scolex narrows considerably to pass directly into the segmented chain. The first two or three proglottides are distinctly broader than long, and the next three or four increase in length but not in breadth, so that the sixth or seventh segment is longer than broad. Mature and gravid segments are slightly broader than long, their dimensions are about 160. antero-posteriorly, 270m across the anterior, and 460m across the posterior borders respectively. But these dimensions are only approximate, on account of the rapid development of the worm. The terminal segment is nearly globular, with an invaginated pore at its posterior end. 410 INTERNAL ANATOMY. Muscular system. The muscle layers are not conspicuous, and as sections were not cut, their detailed arrangement cannot be given. Nervous system. The nervous system was not investigated. Excretory system. The excretory canals, of which only a single one could be seen on each side, lie well towards the lateral borders, and pass ventral to the cirrus pouch and vagina. Genitalia. Testes. The testes are arranged in the two lateral fields of the medulla with the female organs between them. They number about twenty in each segment, and there are usually one or two testes more on the aporal than on the poral side. They lie towards the posterior part of the segment on each side of the ovary (fig. 5). The cirrus pouch is relatively long and runs slightly Fic. 5. A. australis. Mature segments. e.v., excretory vessel; c.p., cirrus pouch ; c., cirrus; t., testes; v.d., vas deferens; r.s., receptaculum seminis; 0., ovary; v.g., vitelline gland; v., vagina. X 90. anteriorly towards the median line. The genital pores are situated about the centre of the lateral border, and are regularly alternating in most cases, but in one or two specimens there were three openings in succession on the same side, which was the greatest irregularity observed. There is no external seminal vesicle, but the vas deferens 411 is thrown into many coils in front of the ovary, before it enters the base of the cirrus pouch (fig. 5). The cirrus is a long tubular structure, and in the one instance observed, where it was extruded, it measured 120” long and 18m thick. It is thickly covered with spines, and has a slightly swollen tip. Ovary. The ovary lies in the median axis towards the posterior part of the proglottides. It is a compact, oval body; its long axis is transverse, measuring about 75y (fig. 5). Vitelline glands. The vitellarium is small and lies transversely behind the ovary, at the extreme posterior part of the segment; in full development it measures about 40 long (fig. 5). The ovary and vitelline glands disappear very suddenly, being apparently in full development in one segment, and almost totally absent in the succeeding one. Vagina. The vagina is a narrow tube running posterior to the cirrus pouch. It expands in front of the ovary into a small receptaculum seminis. Details of the shell gland could not be made out. Uterus and eggs. The uterus was not fully developed, and its mature characters cannot be detailed, but it had the appearance of an irregular sac loosely packed with eggs, and is only visible in at most the three terminal segments. No free eggs were seen, consequently their size is not known. DIAGNOSIS. This interesting cestode closely resembles Angularia beema, Clerc (1906), but differs from the latter in the following points :— (1) Its smaller size (A. beema measures 45 mm. long), and (2) the presence of cuticular expansions on the suckers (absent in A. deema), It is, therefore, proposed to name this species Axgularia australis. The discovery of this cestode is of considerable interest, because the only other member of this genus hitherto described is A. beema, which, according to Liihe (1910), is very restricted in its distribution, only having been recorded from Russia. Type specimens of this new species are in the Museum of the Liverpool School of Tropical Medicine. The genus Angularia, Clerc (1906), resembles closely the genus 412 Gyrocoelia, Fiihrmann (1899), so far as the head is concerned._ Apparently, Linstow’s genus Brochocephalus (1906) is a synonym of Fiihrmann’s Gyvocoelia. The points in which the two genera differ may be summarised as follows :— Angularia. Gyrocoelia. Vagina present. No vagina. Testes 20 to 25. Testes few. Uterus without dorsal and Uterus ring-like, opening by ventral pores. a pore dorsally and ven- trally in gravid segments. REFERENCES Crerc, W. (1906). Notes sur les cestodes d’oiseaux d’Oural. Centralbl. Bakt., Orig. Abt. 1. Vol. XLII, No. 5, Oct. 12, pp. 728-730. Jena. Liystow von (1905). Helminths from the collection of the Colombo Museum. Spolia zeylanica, Colombo. : Line, M. (1910). Die Siisswasserfauna Deutschlands. Cestoda. Heft. 18, p. 115. Jena. 413 NOTES ON ULCERATIVE GRANULOMA BY P. A. MAPLESTONE (Liverpool School of Tropical Medicine) (Received for publication 17 November, 1921) The following notes on cases of Granuloma among Australian aborigines may be of interest, as they present some rather unusual features. The patients were treated at the Australian Institute of Tropical Medicine, Townsville. CASE 1. Male, aet. about 50. The disease was first noticed about two years before admission to hospital; it began on the usual site, viz., the penis, and spread slowly and continuously. After this lesion had existed for some months, an ulcer appeared under the chin in the fold of the neck. When first examined, the case presented the following appearance. The ulcer, which began on the penis, had spread until it reached from one anterior superior iliac spine to the other, and the penis itself had sloughed completely away, leaving the urethra to open in the middle of the ulcerated surface. The ulcer on the neck was about one and a half inches wide, and extended across under the chin, from one angle of the jaw to the other, the sub- maxillary lymphatic glands on both sides were considerably swollen, with consequent difficulty in opening the mouth. The two lesions were similar in appearance, having clear-cut edges and shallow bases; they did not exhibit any granulomatous masses on their surfaces. Diagnosis in both ulcers was established by the discovery in them of the bodies described in Ulcerating Granuloma by Aragao and Vianna (1913). This case presented the uncommon condition of a secondary focus of disease at a site distant from the primary, whither it had been most likely carried by the patient’s hands. CASE 2. Female, aet. 22. About three months prior to 414 admission to hospital the woman noticed a small nodule on the inner surface of the right upper lip; the nodule soon ulcerated, and the swelling and ulceration spread fairly rapidly. When first seen, there was a swelling of considerable size, which extended from the right upper lip, over about the anterior half of the inner side of the right cheek; there was more tumefaction and less ulceration than is generally seen in genital granuloma of the same duration. Smears made from scrapings of this growth were found to contain very numerous organisms, similar to those found in Case 1. Typical infected large mononuclear cell from Case 2. X 1125. There was no sign of a lesion in the genital region, so the case is apparently one of primary infection in a most unusual position. CASES 3 and 4. These were both young native women. They both had small typical granulomatous lesions on the vulva, which they stated had only been present for two or three weeks. On examining the hospital records, it was found that both these women had been treated for similar conditions previously, and that one had been discharged fourteen months and the other fifteen months previously, both being apparently cured. A further striking fact, was that on the second occasion the lesions were on exactly the same site as on the first occasion, in both cases. These two may be cases of reinfection after cure, but the similarity in time of the second manifestation of the disease, coupled with the fact that the lesions are both in exactly the same place as on the first occasion, makes it appear that they are recurrences after apparent cure. 415 TREATMENT The routine treatment adopted in all cases was the application of a simple, non-irritant, antiseptic dressing to the lesions, and the intravenous administration of tartar emetic; I grain in 10 c.c. of normal saline twice weekly. Larger doses were unnecessary, improvement being observed after the first or second injection, and healing continued without signs of relapse during the course of treatment. The time required for complete disappearance of the lesions was proportionate to the extent of the ulceration present; e.g., the first case cited above, with two large ulcers, took about three months to heal, whereas Case 2, with a small amount of ulceration, healed in about a month. On account of the tendency of the disease to recur, five or six injections were given after the ulcer had quite healed. REFERENCE Aracao, H. de B., and Vianna, G. (1913). Pesquizas sobre o Granuloma venereo. Mem. do Instit. Oswaldo Cruz, Vol. V. Rio de Janeiro. ie ed SOirG3 ¥ ‘ST } bo rgeinal atiot enigent “i beyenge reat) x lo:.o:c2priastoming 7 oer vasa eae! oe ture (elesogenail ora sheoh jews t a acetal aiiw Pe hag ganas at brew 12 10 tay ork) yailie heute oils hx aettior of ah -¢syales Tapia tieoitaew, i. cn » tore Jigar a vitor (© se. ened pied i on + : . THIMTARRE te See 1 Kqqm sdfiieuewna fle at basing eBloat + feds, Rolo eeu th ebigees th bipst sat Lintaslo. dae hy les EnMae Stenhoaere sais sek owl alte. aoda - bolts oil ad) {inbre a aee a sot 2 DIM _ ISS OF) SSRSA halaod oats a Wi eee 4 + voalke Goeior Poles <> f A bates” vw my, J ree (a ae Lanne . % ; ‘ * ju! Leaereearties 417 THE STRUCTURAL DIFFERENCES IN THE OVA OF ANOPHELES MACULIPENNIS, A. BIFURCATUS AND 4A. PLUMBEUS BY MALCOLM E. MacGREGOR Wellcome Field Laboratory, Wisley, Surrey. (Received for publication 19 November, 1921) PLATE XXII CONTENTS i. InTRopUcTION ae fi = eae 535 eA ee ave 08 417 - ii. Certarn CHaracters oF Mosquito Ova aie A tee es ac 418 iii, Tue Ova or Anopheles maculipennis 0. ese nee nes tay biases 419 iv. Tue Ova or Anopheles bifurcatus... ... nos MES Pea Ie ie tne 421 vy. Tue Ova or Anopheles plumbeus ... AF sah oe Bae ¥en Hh 421 vi. FAcTs CONNECTED WITH OviposiTIOoN oes aus we see oes wes 422 1. Anopheles maculipennis. 2. Anopheles bifurcatus. 3. Anopheles plumbeus. vii. REFERENCES ... fe ai ees oe nt Sia A ie ose 424 INTRODUCTION During investigations which entailed the breeding and rearing of the three species of British Anophelinae at the War Office Laboratory at Sandwich, Kent, in the summer of 1919, I noticed the quite different appearances between the eggs of Anopheles maculi- pennis, A. bifurcatus, and A. plumbeus, perceptible even to the unaided eye. There are numerous illustrations of Anopheline ova in the English mosquito literature, but I have seen none indicating the markings on the dorsal surface of the ova of A. maculipennis which make it so easy a matter to identify the ova of this species from those of A. bifurcatus. 418 CERTAIN CHARACTERS OF MOSQUITO OVA The eggs of mosquitoes, although showing much difference in form among the various species, nevertheless have certain characters that are uniform. Generally they are elongated, roughly cigar- shaped, or oval objects. The living elements in the eggs of insects are protected by three separate external coats: (1) a delicate innermost coat termed the vitelline membrane; (2) the hard, shell-like material termed the chorion; and (3) a thin outermost membrane which I shall refer to as the enveloping membrane. All mosquito ova have the egg-shell proper (chorion) covered or partially covered with the delicate semi-transparent and water-proof enveloping membrane. This membrane envelopes the egg but is not closely adherent to the shell, a layer of air often existing between the shell and membrane.* Eggs may be easily freed from the membrane by means of pressure with the point of a blunt needle, or by boiling the eggs in water for a moment or two. This membrane may either conform to the surface of the egg and he so close to the egg-shell as to be only visible when special methods are adopted to demonstrate its presence, or it may, by being ‘ballooned’, form structures around the egg. Such structures are seen in the ‘ floats’ of Anopheline ova. The eggs of mosquitoes may be divided into two classes: (1) those that are laid on the water singly, and (2) those that are laid so that the eggs are adherent to each other and float in the form of a raft. In the case of the single eggs these float with their length parallel to the water-surface, and in the case of the raft-eggs they float in a vertical position. In both classes the individual eggs often have a higher specific gravity than water, and consequently not being particularly buoyant, if submerged and the adherent air detached, they sink to the bottom. The membrane, owing to its water-proof properties, acts as an attachment to the water surface film, causing the egg to float as a dry sewing needle may float when it is placed so gently upon the water that the surface tension of the film is not overcome by the weight of the needle. Naturally, the substance of the egg weighs far less than a proportionate bulk of steel, and the stability of the * According to Nicholson the membrane has a highly specialised attachment to the chorion. 419 egg at the surface is thus far greater even in those eggs unprovided with ficats. Nature has apparently recognised that in certain species a greater stability is needed than that afforded to single floating eggs and has developed the plan either of massing the eggs together to form a boat-shaped raft, or by ballooning the enveloping membrane in the single eggs to form the so-called ‘ floats’ so commonly found among the ova of the Anophelinae. The ‘ floats,’ however, do not act to any great extent as true floats giving buoyancy to the eggs, but support them on the surface film securely by presenting a larger area on which the weight of the eggs may be distributed against the surface tension. This fact may be demonstrated by forcibly submerging the eggs, when it will be found that they will either sink or very slowly rise to the surface, generally failing to regain their original position. Mosquito eggs that are laid in raft-formation are almost impcssible to submerge without breaking up the raft form and detaching the eggs separately, on account of the air cells formed between the adjacent sides of the eggs. All mosquito eggs are submerged with difficulty, but submersion can be accomplished with the aid of a fine camel’s hair brush by simply pressing on the eggs with the brush until each egg is driven below the water surface. THE OVA OF ANOPHELES MACULIPENNIS The average dimensions of the ova of A. maculipennis are, length 0775 mm., breadth across the widest part of the float 027 mm. Eggs from different batches vary in their dimensions slightly, but the average variation is remarkably small. When the eggs are viewed from above while they are floating normally on the water it will be seen that the longitudinal outline of the dorsal surface is roughly cigar-shaped and the floats:which lie on each side give the egg a slightly waisted appearance. The dorsal surface is comparatively flattened, while the lower or ventral surface of the egg is evenly rounded. One end of the egg is distinctly blunter and broader than the other. On the ventral side, just below the extremity of the broader end, the micropyle is situated, and it is from the broader end that the larva emerges. The floats are placed 420 on either side of the egg almost at the middle point, though they are actually a little nearer the more pointed end and extend for rather more than a third of the egg’s total length. The float membrane is finely striated transversely, but the striations do not indicate septa dividing the floats into compart- ments. The membrane which forms the float is superimposed upon the main enveloping membrane, being attached to the main envelope only along the ventro-lateral edges. This separate sheet of membrane which forms the float is ballooned outwards and curved upwards until it touches the dorso-lateral surface of the egg, to which, however, it is not adherent. By submerging an egg it can be viewed from several aspects. Laid on its side it presents the outline shown in Plate XXII, fig. 1 (6), and upside down it has the outline shown in fig. I-(¢). It will be noticed that the under surface of the egg is somewhat boat-shaped and draughts progressively less towards the extremities of the egg; also that the enveloping membrane on the under surface of the egg shows hexagonal markings. The enveloping membrane embraces the egg completely except at the micropyle, where a small, round, black area is visible on the under surface of the blunter end of the egg. When the upper surface of the egg of A. maculipennis is examined it will be seen that the enveloping membrane spreads out in areas of unequal thickness, presenting, in consequence, dark and light markings as shown in fig. 1 (a). In these markings I find a very easy method of distinguishing the eggs of A. maculipennis from those of A. 4z/urcatus (compare figs. 1 (@) and 2 (@)). The markings are easily seen. with a hand lens x 6, and to the unaided eye give the eggs of A. maculipennis an ash-grey hue, whereas the eggs of A. dzfurcatus, having no mottled markings on the dorsal surface, appear almost black in colour. The advantage of being able to distinguish the eggs of these two species from each other is considerable, especially when the species are being bred in the laboratory. Only one cage need be used, in which both species may be kept together; and oviposition may be allowed to take place in the same dish. The ova are then easily separated. 421 THE OVA OF ANOPHELES BIFURCATUS Average length 0°61 mm.; breadth across the widest part of the float o'19 mm. In general outline and structure the ova of A. bifurcatus closely resemble those of A. maculipennis, but the enveloping membrane which cradles the egg to the water line does not extend over the dorsal surface, and the dense black egg-shell is naked. The floats in this species overlap the lateral edges of the egg and give it a distinctly waisted appearance between the floats ; moreover, the floats are often of unequal size, and asymmetrical. The two ends of the egg taper to a nearly equal degree, and the curvature of the ventral surface is greater than in the case of the eggs of A. maculipennis, causing the ends of the eggs to be more conspicuously upturned. THE OVA OF ANOPHELES PLUMBEUS The average measurements are, length 056 mm.; greatest breadth 0°17 mm. It has been found that to get A. plumbeus to tolerate laboratory conditions is far more difficult than with A. maculipennis and A. bifurcatus. It was only after numerous attempts that I finally succeeded in getting ‘wild’ specimens to Oviposit in the laboratory. These specimens were captured in Epping Forest by exposing ourselves to attack and allowing the mosquitoes to feed. When replete, the mosquitoes were secured by placing a glass tube over them as they rested upon the skin of the arm. They were then transferred to the laboratory and placed in large insect cages containing an almost moisture-saturated atmosphere at a temperature of 30°C. Porcelain photographic developing dishes containing the water from beech tree-holes on which a few dead beech leaves floated were placed in the cages. Four or five days afterwards the ova of this species were found floating on the water contained in one of the dishes. Owing to the colour of the tree-hole water and the form of the eggs they were exceedingly difficult to see, and when viewed as they floated on the deep brown-coloured water the ‘ floats’ were quite invisible. The eggs were carefully transferred to clean tap-water, and were then seen to be particularly beautiful objects (Plate XXII, fig. 3). _Unlike the ova of A. maculipennis and A. bifurcatus,; the ova of 422 A. plumbeus are somewhat like the ‘diamonds’ of playing cards in outline, and are completely surrounded by the floats. The eggs are more pointed at the ends, both ends being alike, and bend slightly upwards. The central part of the upper surface is also raised, and the lower surface is evenly rounded. It will readily be seen from the illustrations accompanying this paper that the ova of A. maculipennis, A. bifurcatus, and A. plumbeus are so conspicuously different from each other that the differences afford an effective means of identifying the species. FACTS CONNECTED WITH OVIPOSITION Anopheles maculipennis. There is no difficulty in getting the females of this species to oviposit in the laboratory. Specimens captured in pigsties and placed in a large insect cage in the laboratory live well, and will lay several batches of eggs if a guinea-pig is placed in the cage overnight two days after the first and subsequent batches of eggs are laid. The eggs are always laid during the hours of darkness; most often, I think, in the early morning hours before sunrise. I have once obtained a batch of ova at 12 o'clock noon by placing the cage in a dark room. The eggs when first laid are white, but they usually darken rapidly. Sometimes individual eggs in a batch will, however, remain white and fail to hatch, due, I think, to non-fertilisation. This condition is most readily seen in the ova of females that have been kept in the laboratory for some time and have laid numerous batches of eggs, and it is probably to be explained by a diminished supply of spermatozoa in the spermathecae. The average batch of eggs laid by A. maculipennis in the laboratory has been found to be about one hundred and fifty. Anopheles bifurcatus. The eggs of this species are more difficult to obtain, for two reasons. Firstly, the adults are only numerous in many localities between the months of March and the end of May; and secondly, while I have found it easy to get oviposition to take place in the laboratory in the early months of the year, yet specimens captured 423 with difficulty after the end of May and placed under similar conditions cannot be induced to lay, even after they have fed and the ovarian eggs are fully developed. Differences in atmospheric temperature seem to account for this, as.I find that as soon as the weather becomes cooler in the autumn and the mosquitoes become more numerous again, the females will readily oviposit. The average batch of eggs from A. 0z/urcatus in the laboratory has been found to be smaller than that of A. maculipennis, comprising about one hundred and twenty. Anopheles plumbeus. This species has been the most difficult from which to obtain ova. As mentioned previously, repeated attempts had to be made with fed specimens captured in Epping Forest before I was finally successful. So far, I have only managed to obtain three batches at different times from two females. In each case the batch was quite small, consisting of from fifteen to twenty-one ova. With one of the females the second batch was laid a day later than the first, and this is probably a usual practice with the species to avoid overstocking any one breeding-place, since the majority of tree-holes have only a small water-holding capacity, and comparatively few larvae are to be found in each. I have recently extended my observations to the ova of the British culicine mosquitoes, and find that in the ova of the species that I have so far examined there are remarkable specific differences sufficiently marked to be seen with the aid of a hand lens x 6, and to enable the identification of the species to be easily made. On this _ subject a further publication will be made shortly. Probably owing to the difficulty in getting the majority of the species of mosquitoes to oviposit under laboratory conditions, there is a distinct scarcity of information on the specific differences in the ova. Most of the work on the biology of a species in the existing literature begins with descriptions of the larvae in the first instar, and ends with descriptions of the adult and its life history. The egg stage is, nevertheless, quite as important as any of the life stages—in many ways, from a practical point of view, a particularly important stage. Nuttall and Shipley (1901 and 1903), James and Liston (1911), and Howard, Dyar and Knab (1919) are noteworthy 424 in having given special attention to the description of certain Anopheline ova, but considering how numerous are the species of the Culiczdae, it may be said that study and description of their ova has been almost neglected. REFERENCES Gitrs, Geo. M. (1902). ‘A Handbook of the Gnats or Mosquito.’ J. Bale Danielsson & Sons, London. Howarp, Dyar and Knas (1919). ‘The Mosquitoes of North America and the West Indies.’ Smithsonian Institute, Washington, D.C., U.S.A. Vol. I, p. 140, Vol. II, Plates 145-7. James, S. P., and Liston, Gren W. (1911). ‘A Monograph of the Anopheline Mosquitoes of India,’ p. 33. Mrrcuett, E. C. (1907). ‘ Mosquito Life.” Putnam & Sons, New York and London. Nicnorson, A. J. (1921). ‘On the Development of the Ovary and Ovarian egg of a Mosquito Anopheles maculipennis, Meig. Quarterly Fournal of Microscopical Science. Vol. 65, P. III, pp. 395-448. Nurratt and Surprey (1go1 and 1903). ‘Structure and Biology of Anopheles.’ ‘fournal of Hygiene, pp. 49-81 and Plate II. Vol. XV, p. 424, to List of References add : Curisropuers, S. R. (1916). An Indian tree-hole breeding Anopheles, A. barianensis, James = A. (coelodiazesis) plumbeus, Haliday. Ind. Fourn. Med. Res., Vol. TU, p- 489. Eysetr, A. (1912). Cyclophorus (Anopheles) nigripes, Staeger, nov. gen. Archiv. fur Schiffs- und Tropen-Hyg., Vol. XVI, p- 421. ew 426 EXPLANATION OF PLATE XxXIl Fig. 1. Ova of Anopheles maculipennis: (a) dorsal, (6) lateral, (c) ventral aspects of ova. Fig. 2. Ova of Anopheles bifurcatus: (a) dorsal, (6) lateral, (c) ventral aspects of ova. Fig. 3. Ova of Anopheles plumbeus: (a) dorsal, (4) lateral, (c) ventral aspects of ova. Annals Trop. Med. & Parasitol., Vol. XV PLATE XXII a) Z| 2 ee ; — = = = = S cS SS . \Y 2 Fic 3 Fic, 1 Fic. c i aa 427. THE TRYPANOCIDAL EFFECT OF PHENYLGLYCINE AMIDO ARSENATE OF SODIUM ON T. BRUCEI IN RATS AND T. RHODESIENSE IN MICE BY S. ADLER,’ M.B. (Received for publication 23 November, 1921) This drug* is a white amorphous powder readily soluble in distilled water, and yielding in 5 and 10 per cent. concentration a perfectly clear, colourless solution. On standing, however, for some days, a yellow colour develops, whether the solution is kept exposed to light or in the dark. A 5 per cent. solution when exposed to light became yellow in seven days; when kept in the dark, in six days. Daily sterilizing for ten minutes did not prevent the development of the yellow colour, but rather accelerated it. Rats. Minimum lethal dose. In Table I are set out the experi- ments performed in order to ascertain the minimum lethal: dose for rats; in all cases the drug was used in a freshly prepared solution of 5 or 10 per cent., and was injected intraperitoneally ; it produced no irritation. Healthy and infected animals were used in this experiment. It will be seen from the table that no dose was toxic to the animals injected until the amount of 1°2 gms. per kilo was attained. Relationship of Toxicity to age and change of colour of solutions. The number of experiments bearing on this point is rather limited, but serves to show that the toxicity increases on standing, e.g-, although the minimum lethal dose of freshly prepared solution for rats proved to be 1'2 gms. per kilo body weight, an animal which was injected with 018 gms. per kilo of a twenty-four hour old solution followed on the next day by a dose of 0°36 gm. per _ kilo of a forty-eight hour old solution, died after severe symptoms of poisoning on the sixth day after the second injection. In the * Kindly put at our disposal by Messrs. May & Baker, Ltd., Battersea, London. 428 Taste I. a Showing the minimal lethal dose for rats. No. of ob] | Experiment | Dose in gms., perkilo | Infected or healthy Remarks I 0°33 Healthy | No toxic effects 2 0°46 Healthy No toxic effects i 3 0°43 Healthy | No toxic effects rt 0°67 Infected | No toxic effects 1 5 068 Infected No toxic effects | 6 o'7 Infected No toxic effects 7 ; O71 r Healthy No toxic effects 8 o'75 Infected | No toxic effects 9 o'82 Infected ' No toxic effects 10 o"go Infected No toxic effects Bites 1'o _ Healthy | No toxic effects | 12 vi Healthy | No toxic effects 13 12 Infected Animal died in 22 hours Hil | 14 1'4 Healthy | Died on 4th day 429 above solution, although highly toxic, no change in colour was evident. Further experiments proved that in still older solutions the toxicity diminished, e.g., on the eleventh day, when the solution was deeply yellow, three animals were each given a dose, approximately equal to the combined doses given above, without producing any symptoms. The first of these three animals was injected with a solution which had been kept in the daylight and boiled each day for ten minutes, the loss from evaporation being made up by addition of distilled water to the original volume before injection. The second was injected with a solution which was unboiled and kept in daylight; the third with solution unboiled and kept in the dark. The increase in toxicity which was evident in the forty-eight hour old solution, was accompanied by a definite increase in trypanocidal power, for after injection of 0°36 gm. per kilo, trypanosomes disappeared from the peripheral blood within twenty hours, and were absent from the blood until the time of death. Examination for trypanosomes of the organs by smears proved negative, as did also the injection of emulsion of organs and blood into healthy rats. A similar sterilizing effect could only be produced by much larger doses of the freshly prepared drug, e.g., a dose of 0°67 gms. per kilo in freshly prepared solution injected into a rat at approximately the same stage of infection as the previous one, only caused the disappearance of the parasites from the peripheral. blood in forty- eight hours, and did not prevent their re-appearance twenty-four hours later, Toxic effects of freshly prepared solutions. No toxic effect was observed below the dose of 1'2 gms. per kilo; a heavily infected animal died within twenty-four hours of receiving this dose. The signs of poisoning noted before death were blindness and refusal to take food. No gross haemorrhages were found after death, but the whole intestinal tract showed numerous minute haemorrhages. No trypanosomes were found in the peripheral blood, either immediately before or after death. Loxic effects of old solutions. Toxic effects were noted after a dose of 0°36 gm. per kilo, but not until four days had elapsed. On the fifth day the 430 animal appeared ill, lying curled up, breathing irregularly and spasmodically , it refused food, was blind, and staggered in its gait when moved. Haemorrhages were observed from the conjunctiva, anus and urethra. Post-mortem, the whole intestinal tract was found to be haemorrhagic, the liver was enlarged and soft, and showed yellow mottling; the kidneys were very soft, but not enlarged nor dark in colour; the spleen showed yellowish patches. Minimum curative dose. In Table II are sct out the experiments performed in order to demonstrate the minimum curative dose in rats. Taste II. Shewing effect on rats infected with T. brucet. Day of Trypanosomes disappearance of Experi- per field. Dose in gms., | trypanosomes | Day of Remarks ment Ocular 4, per kilo from peripheral | re-appearance Obj. 4 blood I 40 243 - 5 2 Swarming 0°67 2 3 3 Swarming 0°68 I | 2 4 Swarming o'7 I | ade No relapse after 7 months 5 36 O'75 I “ee No relapse after 7 months 6 14 o'82 I ee No relapse after 7 months 7 25 o'gl I one No relapse after 7 months 8 Swarming 12 74 7 Died in 22 hours. No. trypano- somes Control Swarming O17 I 17 (atoxyl) It will be seen from the above table that while the atoxyl control animal was rendered free from trypanosomes in its peripheral blood 431 Taste III. Showing effect on mice infected with T. rhodestense. Io il 12 Trypanosomes per field. Ocular 4, Obj. 4 Swarming Swarming Numerous Numerous Swarming Numerous Swarming Numerous Numerous Numerous Swarming Control | Dose in gms., per kilo o's 06 "74 0°83 o"9 o"9 I’o Vl 1°25 16 20 Day of disappearance of trypanosomes Day of Remarks from peripheral | re-appearance blood 2 4 ° 2 6 2 5 I 8 2 5 I 4 = 5 2 Alive and well 5 months later 2 4 I 6 I 12 2 Z Died after 3 days 432 by o'17 gms. atoxyl per kilo, but relapsed in seventeen days, the experimental animals were rendered free from trypanosomes when a dose of 0'7 gms. per kilo of phenylglycine amido arsenate of sodium was reached, and did not relapse. Action in Vitro. No trypanocidal action in vitro was observed, either by the drug itself or by the serum of animals, twenty-four hours after they had been rendered free from trypanosomes by injection of the drug. Mice. Minimum Lethal Dose. In Table III are set out the experiments performed in order to demonstrate the minimum lethal dose for mice and the effect on T. rhodesiense. It will be seen from the table that the minimum lethal dose was 3 gms. per kilo; the effect on the trypanosomes was negligible up to a dose of 2°5 gms. per kilo, only one animal failing to relapse. SUMMARY Phenylglycine amido arsenate of sodium can be used in freshly prepared solutions in distilled water for intraperitoneal injection into rats and mice; the solutions on standing become toxic, and later become yellow in colour. For rats, the minimum lethal dose of the freshly prepared drug proved to be 1:2 gm. per kilo of body weight. For rats infected with 7. d7wcez, the minimum curative dose is o'7 gm. per kilo of body weight. In vitro the drug has no appreciable action on trypanosomes, nor has the blood of treated animals immediately (twenty-four hours) after becoming trypanosome-free. The drug has no curative effect on mice infected with T. rhodesiense. A remarkable feature of this drug is its relatively high minimum lethal dose. Although the drug contains 26 per cent. arsenic, the minimum lethal dose was found to be 1:2 gms. per kilo for rats and 3 gms. for mice. I am indebted to Professor Blacklock for carrying out the necessary inoculations for the experiments. 433 ‘ NOTE ON BISMUTH AS A TRYPANOCIDE BY S. ADLER, M.B. (Received for publication 23 November, 1921) The effect of bismuth in the form of soluble bismuth sodium tartrate in solutions of various strengths was tried on animals infected with ZT. rhodesiense and T. brucei (Nagana ferox) respectively. Using this drug, the minimum lethal dose for healthy mice was found to be 0'047 grammes of bismuth, and for healthy guinea-pigs 0'062 grammes of bismuth per kilo body weight. In animals which died after injection of bismuth sodium tartrate deposits of bismuth were found in all cases in the liver, frequently in the spleen, and less frequently in the kidneys. T. rhodesiense. The minimal lethal dose cleared the blood of trypanosomes in a mouse within twenty-four hours, but the animal died in two days. Any dose below this failed to clear the blood of trypanosomes. T. brucei (Nagana ferox). Although the drug cleared the blood of trypanosomes in guinea- pigs, yet relapses occurred in a few days, thus :— Dose per kilo, body weight Injected | Trypanosomes re-appeared | oro18 20.4.21 | 28.4.21 07022 26.4.21 | 2.5.21 o°024 | 9.5.21 | 18.5.21 0°032 9.5.21 | 21.5.21 In no case was a cure obtained. I am indebted to Professor Blacklock for carrying out the necessary inoculations for these experiments. ee = Bp eficcion: so one name em Nee ee a 4 fo ‘@ - OO eer . Le EPO ae = t et Tay @ t ih) b egpol 4th 4p tani hi chi (youve aos iay AOE a h f “1 ae 7 + A") Diener Siew bon iontacot) aii + Poni iat . pr ‘ f rasa | gtit rah oie } ves rey Gol fifi en 11 er ¥ 142) ok sine” wash U at efieapent ash Baas pk ——s iy i tay io ee riety geoky (adaah te 7 \ ae iid ‘ath wD ahaa clay ey at 3m (Tat Battal alr : At yg Seo et \> beckon i pare VaR Glare Daten P: ~ ie ~ - a ~~ oe ty « v@i os aed a : . i = hfe ie t wal +): ae ee : Fs Wie: |g Ge ae . Te a - ‘4 fh lat Le eiaalts “ iid _ See YET: ie Beniatte Ks Oa dt A oiticls. eagdon oh De oie ; rr, ingen raed: \ pe 4 435 MALARIA ON A VENEZUELAN OILFIELD WW STEPHENS, UMD!) F.R.S: (Received for publication 5 December, 1921) PLATES XXIII AND XXIV The oilfield is situated on the eastern side of Lake Maracaibo, Venezuela, about 80 miles from the head of the lake, and 10 miles inland from its shores. The following observations were made in the month of August, 1921. MALARIA IN THE NATIVE POPULATION The extent to which malaria prevailed in the native population of various villages in the district is shown in the following table : — Age Period... ec aes o—I0 II—20 21— No. |Percentage| No. Percentage | No. | Percentage exam. | infected | exam. | infected | exam. | infected Old Village snail Waxes Io if) 8 APS chic? _ New Village ay oe 32 468 39 23°0 | 49 Ser San Pedro ... ae a 9 66°6 6 oro I oro Los Barrosos ae ona 9 oo 9 o'0 ° -- San Timoteo gee sso 27 Oy) | 4 o’o ° -- The relationship of malaria in the Old Village and New Village respectively to malaria among the white employees on the oilfield we shall discuss later. The other three villages were not in the immediate neighbourhood of the oilfield, but it may be pointed out here that the absence of infection in Los Barrosos was probably due to the fact that anophelines were only found at a distance of half a mile, and then only in one small clay-pit about 436 12 feet square. San Timoteo is a village built on piles over Lake Maracaibo. Its low endemic index, 3°7 in children under ten, is thus probably due to its position ovér the lake and to the fact that only one small anopheline larva was found after prolonged search on the mainland. San Pedro is four or five miles from the oilfield, and its high rate is probably due to its being situated close to extensive anopheline swamps. SPECIES OF PARASITES FOUND In over two hundred blood specimens examined, malignant tertian malaria (crescents) was only found twice, simple tertian and quartan parasites were about equally common, a peculiarity of infection which I had not previously seen in Africa or India. The frequency with which pigmented leucocytes were observed in the ‘positive’ films was a noteworthy feature. MALARIA AMONG THE WHITE STAFF No precise information was forthcoming as to the nature of the sickness among the white employees (about thirty in number). An examination of the blood~of sixteen employees in apparent good health revealed nothing. In the case of one employee suffering from fever, simple tertian parasites were found. In the absence of definite records, one had therefore to assume that the sickness prevalent among the staff was due to malaria. POSITION OF THE STAFF HOUSES They are situated (vzde Map), (1) on two ridges a few hundred yards from the first source of Anopheles, viz., the small anopheles swamp in the Asphalte creek, the houses on the easterly ridge actually overlooking the swamp. (2) From the second source of Anopheles the ‘Anopheles Swamp’ which begins due west and even north-west ‘of the New Village, they are situated less than half a mile. (3) They are situated less than a quarter of a mile from a source of malaria infection, the Old Village, and (4) since the year 1920 half a mile from a second source of infection, the New Village. We have here then an example of the conditions so frequently er OO HTS yi PLATE Annals Trop. Med. & Parasitol., Vol. XV 2niyooan IAyOOaN (ysis) So}090)Q4, 10d Anijisod xajmy RYO joo ear (ouu i) enrposau a *Pa AAoS au QAI} obeu ve 5 usu) $108. (9 F Ys) SPA" IK(yOOSU ea an \JoSay 5/09) AAIpOOaU aMiyosau yee (not YL) sjoqy DAIyOSeU = asuoyu § uvis BPLOL ui Ajaaay Buijiq x9[7) AP S\BINAKL e DAt|\Sod x0\r) jood \ows fot 2 Duijiq x oe ey eee AAI ~ BAIOOeU Canine ‘Ss ANi}osay y (aiiaye AAINOSN 7 (QuAIWNVO NOOO! 7. Peony) J Fa p yaod payor ear — WNWEWS IN14 AUG ANIAWE kun anihisod xaja5 He MOL LOET Ypsres $ yNyev / j yin ad WNvavs iv1a Aug JNIAWH AUT PAINE ier HPD Alpydsy TIA axyodau duo pete Larue EIR anilisod xajay xv eed Se sjood. may 2 710M ay) Auanyy r nN Ss afoudsy Sniysod Bojan j No sadois apoudsy _jduroms (plus, Yoos [nego a tas hap sadojs aoydsy sadojg, ehjoydsy f sadojs a\oudsy >/ s Ss AQTAld-10 NVT3NZANSA irae OOOO! :1-d1VvoS / Vv JO C. Tinling & Co., Lid., Imp ari ynhat = LO POR Tepe AN\\ eee cde ie mh 1 437 founds in the tropics, namely, native villages and a source of Anopheles in close proximity to a white population, and here, as elsewhere, the conditions would completely explain the existence of malaria among the white employees. THE ANOPHELINES The district under investigation can be divided into four areas :— : A. The Jungle. B. The cleared area. C. The residential and Old Village area. D. The New Village area. The relative positions of these will be seen on the map. AREA A. THE JUNGLE. Numerous examinations were made of all discoverable water in the jungle so far as it was penetrable. In many cases, pools and streams and ‘tatucales,* apparently favourable breeding-places, were examined with negative result, often, no doubt, due to the presence of abundance of small fish (in some cases due to a covering of Lemma (duckweed). In other cases, however, jungle pools were ‘Culex’ positive. One interesting example was encountered, viz., that of a borrow- pit, where the snouts of dozens of small fish could be seen voraciously hunting the surface for food, and yet ‘Culex’ larvae in abundance were found in the protecting weed and rubbish at the sides, but none away from there. Again where the jungle was apparently dry over large areas, yet ‘ Culex’ was present and bit freely in the daytime when one stood still. Anopheles (mosquitoes or larva) was, then, absent from the jungle at least in August, and mosquitoes caught in the jungle at long distances from the oilfield were invariably ‘ Culex.’ AREA B. THE CLEARED AREA. Here and there a few ‘Culex’ breeding-grounds were found, generally in small numbers. A swamp a hundred yards or so long, * Tatucales: Diminutive islands or mounds intersected by water, and in which progress could only be made by a series of jumps. 438 passing in front of the native mechanics’ quarters 1 to 6 and close to the next area, is of interest in that repeated search showed it to be Anopheles negative, but it contained a small ‘Culex’ area near its origin. It had been already partly, but incompletely drained, and fish abounded. AREA C. STAFF HOUSES AREA. The Staff Houses are situated for the most part on two ridges themselves separated by a dry ravine. These ridges are bounded on the N.W. and N.E. by the Mene Grande and Asphalte creeks respectively. These creeks merge at their origin northwards, and are bounded by the slopes of the ridge on which the Star Water tank is situated. Again further north to the west and the east this ridge descends in sloping ground, dry for the most part and covered with asphalte until in a N.W. direction the creek ‘ Piedritas blancas’ is reached. The whole of this large area is for the most part dry. What little water there is, is found in the slopes draining to the road from the Old Village to Zo, but frequent examinations of all collections of water in this area were negative. As regards the Asphalte creek itself, this was also negative except for a small swamp some fifty yards long in its upper portion. Here Anopheles larvae in scanty numbers were found. The rest of the creek had already been drained; the ditches contained numerous fish and considerable quantities of oil. The swamp above referred to was caused by a small depression in the surface, and was easily abolished by cutting the long grass and filling with earth from the adjacent higher ground. AREA D.—oR NEW VILLAGE AREA. The New Village is situated on the Sabana de Matajey de Raya on elevated ground (thirty metres) a hundred yards or more from the creek El Mene or water creek. From the elevated flat ground on which the village lies a number of dry ravines lead down to a swamp some fifty yards or so in width, bordering the stream formed by the junction of the streams from the Asphalte and El Mene creeks. This swamp extends along the edge of the Sabana in a curved course (probably as far as San Pedro), and was found to contain Anopheline larvae—always, it is true, scanty in number—for a distance of, 439 roughly, abcut three-quarters of a mile. Although fish were plentiful in the two drains that had already been cut in part of it, but which were blocked when examined by me, yet Axopheles were present always in small numbers, but from its extent this swamp and its tributary swamps (vide infra) formed the main source of Anophelines. YVhe Tatucales, out in the Sabana, contained fish, and was negative. The ravines below the village to which reference has been made, extend in a south-easterly direction, becoming less steep and with flatter bottoms so that they are no longer dry but are occupied by swamps; it is these secondary tributary swamps and the main swamp whick they eventually join, which formed the great Anopheline breeding-ground. In the four areas examined, Axopheles were found breeding in two only, and in one of these only in a swamp of small dimensions. The breeding-grounds were thus in August definitely restricted ; Anopheles were not breeding in the jungle nor in the open cleared area, nor in the waters that existed on the asphalte slopes of Area C. The swamp in which Anopheles were found breeding was over- grown with a variety of grasses, some up to 6 ft. high; in other parts it more resembled marshy ground with short rushes in part trodden by cattle; in other parts again ‘tatucales’ formation. In no case were Anopheles found amidst overhanging trees. Anopheles (C. argyrotarsis) were also found breeding in one of half a dozen clay borrow-pits some three miles away from the camp. ADULT ANOPHELINES. The search for anophelines in the native huts in the daytime was completely fruitless, and culicines also were very scanty. This condition was in marked contrast to those observed by me in the neighbourhood of Lake Valencia, which I visited on my way home, where in the daytime, in the verandah of a hut, it was easy to collect numerous anophelines, embracing three different species. On the oilfield itself I was only able to secure by capture at night in the native village sixteen specimens of C. avgyrotarsis, and during my stay in the camp no anophelines and very few culicines were seen by 440 me in the house I occupied. It should be stated that the windows of all the houses in the camp were wire-screened, and mosquito-nets were in general use. PROPHYLAXIS The existence of infection in the Old Village and the absence of Anopheles from any area nearer than an isolated small, grass-grown_ swamp in the Asphalte creek and the great Anopheline Swamp adjacent to the New Village proves, I think, that we have a normal flight range of about half a mile. We may consider prophylactic measures under the following headings :— (a) DRAINAGE OF ANOPHELINE SWAMP. Towards the end of my visit, 100 yards or so had been cleared of grass and existing choked drains cleared and additional ones constructed, with the result that the swamp rapidly became dry and free from larvae in this section. I suggested that the swamp should be drained for about a mile. The efficacy of this measure can be estimated by observing the effect (1) on the number of anophelines caught in the New Village month by month; (2) on the endemic index of the New Village; (3) on the sickness rate of the white staff. If the drainage results in a complete or almost complete suppression of anophelines, further measures would hardly be necessary, but in the event of this not being so, other measures would have to be adopted. (6) REMOVAL OF THE OLD VILLAGE. This source of infection in close proximity to the white staff quarters should be abolished, and if further protection is sought various stray native huts which exist here and there should also be removed. (c) THE NEW VILLAGE. If drainage of the anopheline swamp is unsuccessful, it would be necessary to consider the question of removal of the New Village to an area free from anopheline breeding-places; such a one existing 441 a mile or so from its present position. For elimination of malaria from the village itself, quinine administration is advisable; and further, if it remains in its present position wire screening of the huts could be advantagéously employed, without prohibitive cost. (dz) A more fundamental and expensive procedure would be the removal of the white staff houses. An ideal site for these exists on the Star Tank ridge, at an elevation of about 300ft. and at a distance of about a mile from the New Village and Anopheline _ Swamp. One sees, accordingly, that the existing conditions have arisen from a lack of appreciation of the fundamental principles responsible for the infection of the white man amidst a native population, namely, close propinquity to that population, with a supply of anophelines also adjacent. Had one been laying out the camp de novo, the following arrangement would, I believe, have led to the protection not only of the white but also of the native population :— (1) The white staff houses should have been placed on the Star Tank ridge; (2) the native village should have been placed a mile from this site and also a mile from the source of anophelines. The conditions prevailing were particularly favourable for such an arrangement, namely, (1) the complete absence of anopheline breeding-grounds over a large part of the area considered, and (2) the large tract of ground in which collections of surface water of any sort were absent (in August). NOTES The oilfield was situated in Lat. 9°.75/ N. and Long. 71°.15/ W. The indoor temperatures in August ranged from 85°-89°5° F. maxima to 75°-81'5° F. minima. Heavy rain lasting about half an hour fell towards evening on five out of twenty days. The only species of Anopheline found in the area was Cellia argyrotarsis, distinguished by having three and three-quarters hind tarsi almost completely white. The palmate hairs of the larva of this species are variable. Usually there are palmate hairs on segments two to seven, that on segment two being small and hard 442 to see. Examples were also found with well developed palmate hairs on segments one to seven, together with a well developed thoracic palmate hair. Examinations of sixteen specimens of C: a7gyrotarsis for sporo- zoites proved negative, but Tovar states that this species transmits in Venezuela. The small fish which abounded in the small streams and in the swamps of the district belonged to the following species :— Gambusia (Poecilia) tridentigera, Hapfplochilus sp., and Chromides (Acara) dorsigera. he Ve 4 5 7 = -wolsone to ‘say rebolt gyre a. at e AS oy ; igre ; : yo j Sige} ; © t Fim aa vos 4 tC sais . eta 4 Fk i. 7) he a te at Z mbes od T soe Rave do wis Ses bid t iy . Se [ek bit ord a =p osttosehs Sis mie Toad Yl ‘ - OF Iniog ewann scl saeogolemenace oC sy: L mee } % - Fig. Fig. EXPLANATION OF PLATE XXIV Old type of Bungalow. Modern type of Bungalow. The Peons’ Cock-pit. Bird’s-eye view of Staff houses. The ridge with Star Water Tank in the distance. The Asphalte slopes. The arrows point to two asphalte mounds. Annals Trop. Med. & Parasitol., Vol. XV s PLATE XXIV Fic. 4 Fic. 5 C. Tinling & Co., Ltd., Imp. 445 NOTES ON CULICIDAE COLLECTED IN VENEZUELA BY Ae MS VE VANS: “M:Se (Received for publication 25 November, 1921) PLATES XXV AND XXVI During the course of the investigations in Venezuela, recorded in the foregoing paper, a number of mosquitoes were collected by Professor Stephens at Mene Grande, which is situated about ten miles inland from Lake Maracaibo. Specimens were also caught at Maracay near Lake Valencia, which is separated from Lake Maracaibo by a range of mountains, and on the Island of Curagao. The following is a list of the species collected at each of the three places :— LAKE MARACAIBO Anopheles argyrotarsis, R.D. Hatched from larvae taken in swamp 3¢ 3, 226. Mene Grande, 22.8.2, 99 5. ; Aedes serratus, Theo. Mene Grande, 3.8.21, 99 6. Biting in jungle, Mene Grande, 5.8.21, 99 6. Caught in jungle, biting horse and man, 5.8.21, 21; 11.8.21, 92 11. ey Aedes scapularis, Rond. Mene Grande, jungle, 1.8.21, 29 3. Taeniorhynchus titillans, Walk. Mene Grande, biting in jungle, 29 3. Culex (Neomelanoconion) chrysothorax, Newstead and Thomas. Reared from pupa taken in swamp, Mene Grande, 7.8.21, 9 I. 446 *Culex (Culex) coronator, D. and K. Mene Grande, 6.8.21, 33 4, 22 6. Hatched from larvae, Mene Grande, 8:21, 63 2; EL 6:2, 292.) dd 3) Oe *Culex (Culex) nigripalpus, Theo. Mene Grande, ¢ I. Psorophora posticata, Wied. Mene Grande, 11.8.21, 29 2. Joblotia digitatus, Rond. Jungle, Mene Grande (about 20 miles inland from Lake Maracaibo), 20.8.21, 22 2. MARACAY (about 10 miles from Lake Valencia) Anopheles argyrotarsis, R.D. Maracay outskirts, 8.9.21, 2 I. Anopheles albimanus, Wied. Maracay outskirts, 8.9.21, S I, QI. Anopheles albimanus var. tarsimaculatus, Goeldi. Maracay outskirts, 8.9.21, ¢ I, 2 1. Maracay outskirts, 5 specimens. Anopheles pseudopunctipennis, Theo. Maracay outskirts, 7 specimens. Aedes scapularis, Rond. Maracay outskirts, 8.9.21, 9 I. Aedes trivittatus, Coq. Hoan, (eh 7. *Culex (Culex) quinguefasciatus, Say. Maracay outskirts, 8.9.21, 33 3. *Culex (Culex) virgultus, Theo. Maracay outskirts, 8.9.21, 3 I. Psorophora posticata, Wied. Maracay outskirts, 8.9.21, 2 I. * Determined by the male genitalia (See Dyar, 1918). 447 CuRACAO Aedes (Stegomyia) fasciata, Fabr. Breeding in tub, Ice Factory, Curagao, 1.9.21, 2 I. *Culex (Culex) quinquefasciatus, Say. From larvae breeding in tub of ice manufacturer, 30.8.2I, HIS Oy - -@ LEL; Leger odrs14,) | SY LZ e THE MORPHOLOGICAL CHARACTERS OF ANOPHELES ARGYROTARSIS, R.D., AND A. ALBIMANUS, WEID. Larvae. In their Monograph, Howard, Dyar and Knab (1917), state that palmate hairs occur only on abdominal segments two to seven in A. argyrotarsis, but that in A. albitmanus there is an additional small pair on the first abdominal segment. The larvae of A. argyrotarsis, which Professor Stephens collected from the swamps around Mene Grande, however, all possessed these structures on the first segment of the abdomen. In some specimens these were in a reduced and incomplete condition, but in a certain number they were well developed, and in these latter specimens a pair of palmate hairs was also present on the thorax, a position in which they do not appear to have been recorded hitherto in either of these species. The distribution of these structures is thus shewn to be’ variable in the two species, and cannot be used as a specific character for determination of the larvae. Male hypopygium. In view of the close resemblance between the adults of these species, it appeared desirable to compare the detailed structure of the male hypopygium. The figures in the Monograph of Howard, Dyar and Knab (1912) tend to exaggerate the differences present, and it is obvious that the morphology was not clearly understood. Christophers (1915) described the hypopygium of A. albimanus, and placed in one group with this species A. argyrotarsis, A. tarsimaculata, and A. bellator. This group was based on the number and arrangement of the large spines arising from the side-piece, but Christophers stated that it was imperfectly studied. Edwards (1920) referred to the structure of the mesosome (theca) of A. argyrotarsis, saying that it approached nearest to the simple form seen in Ochlerotatus. “Determined by the male genitalia (see Dyar, 1918). 448 Before comparing the hypopygium of the two species, it is necessary briefly to discuss the structure of certain of the constituent parts. The nomenclature employed is that proposed by Edwards (1920). The mesosome (theca) of A. albimanus has been described and figured as ‘ massive and clubbed.’ This appears to have been due to confusion of the mesosome with a wide median membraneous lobe (fig. 2 A and B, m.l.) which arises from the membrane at the base of the side-pieces on their upper (by rotation) sides. The position of AME. O-| millimeters Fic. 1. ¢ Hypopygium of A. argyrotarsis from below; tenth sternite not shown. cl., claspette ; m., mesosome ; 5.p., side-piece ;_s.s., stalked spine. this lobe in relation to the side-pieces is shewn in the drawings (fig. 2 A and B). Distally it becomes bilobed, and the two halves are produced downwards (towards the tenth sternites) so as to embrace the distal half of the mesosome. So intimately are the distal portions of the lobe associated with the mesosome, that in mounted preparations it is often impossible to distinguish the separate structures. When examined floating in oil under a binocular microscope, however, and arranged so that a terminal view 449° is obtained the mesosome can be seen lying ensheathed on three sides by the median lobe, and its downward processes. The separate parts can then be dissected away and isolated. The form of the mesosome and median lobe afford what appear to be reliable a \ fA dt TG: a, 44 fan f tases by ea ls, SU yuen ty (ud i ae \ Dh) a f i) Bi | O-1 millimeters | Fic. 2. Base of g Hypopygium from above; mesosome and tenth sternite not shown; drawn from specimens macerated in K.0.H. A—A. argyrotarsis. B—A. albimanus var. tarsimaculata. m.l., median lobe; s.p., side-piece. characters for the separation of the species A. argyrotarsis and A. albimanus. The latter species and its variety ‘¢arsimaculata appeared not to differ to any marked degree in the characters of the hypopygium. 75° The Mesosome (fig. 3 Aand B). It was found that the mesosome can be best studied in specimens stained with carbol fuchsin. In both species the halves of the mesosome are elongate plates, each articulating basally with the chitinous pieces (g.6.f.), which represent the parameres and basal plates of other Culicidae. The halves (4) approach each other on the upper side, but below are connected by thin membrane which distally becomes chitinised, forming a rounded apical plate (2.~.) concave above, and membraneous at its margin. O1 millimeters Fic. 3. Mesosome; drawn from specimens stained with carbol fuchsin. A—A. argyrotarsis. B—A. albimanus var. tarsimaculata. a.p., apical plate; 4., half of mesosome ; /., leaflet; p.b.p., plate representing parameres and basal plates. In A. argyrotarsis the halves of the mesosome may be actually contiguous for part of their length, causing the mesosome to be tubular in this region. At the distal extremity of each half arises a flat recurved spine (/.), toothed on its outer side, which is obviously equivalent to the Jeaflets of the more specialised Anophelini. Although the leaflets of other species are usually directed more or less distally, Swellengrebel (1921) figures some recurved ones in A. (Myzorhynchus) barbirostris, vy. d. Wulp var. pallidus, Sw., and in A. (Myzorhynchus) umbrosus, Theo. The greatest breadth of the apical plate is greater than its height above the apices of the halves of the mesosome. In A. albimanus and its variety ¢arstmaculata, the halves of the mesosome (fig. 3B) have only a slight tendency to form a tube. Leaflets are absent, and the greatest breadth of the apical plate is 45% equal to or less than its height above the apices of the halves of the mesosome. The Median lobe. In A. argyrotarsis (fig. 2A), the distal portions of this structure are composed of a number of parallel divisions, the exact form of ewhich is exceedingly difficult to determine in macerated specimens, owing to the extreme thinness of the membrane, and its lability to distortion. There are no long hairs or setae present. In A. albimanus and its variety (fig. 2B), the surface of the median lobe is thrown into a large number of shallow folds distally, these folds involving the lateral descending portion as well as the upper surface. The sides are thickly clothed with long hairs, and a number of very delicate hairs occur on the surface of the basal portion of the median lobe. Side-pieces. No well marked specific characters could be found in the large spines borne on the internal surface of the side-pieces. The pedicels of the stalked spines (fig. 1, s.s.) are slightly shorter in A. argyrotarsis than in A. albitmanus. The claspettes are in the former species broad and not well differentiated from the surface of the side-piece. In this they differ from the separate finger- like condition in which they sometimes occur in A. albimanus (Christophers, 1915). In some specimens of the latter species, however, they occurin the same condition.as in A, argyrotarsis. The ¢enth sternite is almost identical in the two species. The appearance varies according to the position of the paired chitinous arms. The main differences, then, between the male hypopygium of A. argyrotarsis and A. albimanus lie in the form of the mesosome and the membraneous structure here referred to as the median lobe. The character of the mesosome of A. argyro¢arsis, with its single pair of leaflets and tendency to form a complete tube, suggests a transitional stage between the generalised condition seen in A. albimanus and the completely tubular form with numerous leaflets which occur in most species of the genus. REFERENCES Curisroruers (1915). Ind. Yourn. Med. Res. Vol. III, p. 379. Dyar (1918). Ins. Ins. Mens. Vol. VI, p. 94. Plates III and IV. Epwarps (1920). Ann. Trop. Med. & Parisitol. Vol. XIV, p. 23. Howarp, Dyar and Knap (1912), (1917). Mosquitoes of North and South America and the West Indies. Vol. II, Figs. 258, 263, 264. Vol. IV, Table on p. 967. SweLtencreset (1921). Overged, u.b. Tijds. v. Ent., Deel LXIV, 1921, pp. 39, 40. 452 EXPLANATION OF PLATE XXV Fig. 1. Wing of Anopheles albimanus var. tarstmaculata, Fig. 2. End of hind tarsus of A. albimanus var. tarsimaculata. Fig. 3. End of hind tarsus of A. argyrotarsis. Annals Trop. Med. & Parasitol., Vol. XV Sie REATE XEN | O°5 millimeter | Iter 9 Bic. 3 1 millimeter Bie. A.M. Evans, ad. nat. del. D Penltan ft Ga Tad tank — — - TAA ae At A Fe ae ” = 7 vp ae rik be i) * . 71 es x ‘ ' # ale ' ’ q Ven ‘ ‘ _ ” 5 % pf 4) 454 te R: by hi 4 al Nie = \ - . = . EXPLANATION OF PLATE XX) Fig. 1. Wing of Axopheles punctipennis. ale ‘’ & ee Fig. 2. Wing of A. pseudopunctipennis. = vy . = al are . a % »~ Me PLATE XXVI XV Rey LE Parasitol Annals Trop. Med. & 1 millimeter IG. A.M. Evens, ad. nat. 455 A NOTE ON THE SYNONYMY OF THE GENUS ZSCHOKKEELLA, Ransom, 1909, AND OF THE SPECIES Z. GUINEENSIS, (GraHaM, 1908) BY T. SOUTHWELL AND P. A. MAPLESTONE (Received for publication 29 November, 1921) In Simpson’s Report on Plague in the Gold Coast in 1908, Graham briefly described a parasite from Cricetomys gambianum, to which he gave the name Davainea (guineensis, n.sp. ?). We have obtained from the Gold Coast on several occasions, through the kindness of Dr. J. W. S. Macfie, large collections of worms, both from C. gambianum and Epimys (Mus) rattus. The examination of this material has led us to make the following observations :— Beddard (1911, a), apparently unaware of Graham’s paper, redescribed the same worm under the name Thysanosoma gambianum. Beddard (r911, b) changed this name to Thysanotaenia gambiana, and again (1912) to Zschokkeella gambianum. Baylis (1915) described Z. muricola, n.sp., from Epimys (Mus) rattus, and Meggitt (1921) described a new pra of Inermicapsifer (I. zanzibarensis), from C. gambianum. We have been unable to find any constant or adequate points of difference between our material and the above three species; this is clearly indicated in the table below. For these reasons we have come to the conclusion that the worm Z. guineensis has the following synonymy, viz.: Davainea (guineensis, n. sp. ?) Graham, 1908 ; Thysanosoma gambianum, Beddard, 1911 ; Thysano- taenia gambiana, Beddard, 1911; Zschokkeella gambianum, Beddard, 1912; Zschokkeellamuricola, oe 1g15; and Inermicapsifer zanzibarensis, Meggitt, 1921. We agree with Meggitt (1921) that the genus Thysanotaenia should lapse, and that no valid distinction has yet been made between the genera Zschokkeella and Inermicapsifer. We therefore suggest that the genus Inermicapsifer, Janicki, 1910, be likewise suppressed, and all the species in it included in the genus Zschokkeella, Ransom, 1909, on account of its priority. 456 .| About the middle. Note—Shown anterior in figure. Genital Pore ... Z. gambianum, Beddard Z. muricola, Baylis Anterior. Testes ... In two separate groups. (This is only implied in the description and is not definitely stated). Vesicula seminalis ? Present. If so, repre- sented by a pouch near the ovary. Ovary ... -| May be regarded paired; partly separ- ated by yolk gland. as | In two groups, but almost continuous. I, zanzibarensis, Meggitt In anterior quarter. Continuous; but fewer in centre of segments. Not seen. Small and inside cirrus pouch. Single, fairly compact, | Single, made up of strag-_ lobulated, crescentic. gling lobes, long, slender and only slightly connected. Receptaculum seminis | Long and not swollen. No. of eggs in capsules | A few. was kind enough to send us. Bayuis, H. A. (1915). A new Cestode of the Genus Zschokkeella. Ann. & Mag. Nat. Hist., Ser. 8, Vol. XVI, London. Bepparp, I’. E. (1gtta). I. On some Mammalian Cestoidea. (1911b). much Small rounded organ near ovary. Large and thin walled. Note—This structure appears to be what is usually considered as the vagina. About 20. REFERENCES II. On two new Genera of Cestodes from Mammals. —— (1912). taenia and Hyracotaenia. Vol. XIII, No. 3, Cambridge. One, but g to ro eggs in clusters. * Diagnosis compiled from observations on our material and from specimens which Dr. Meggitt Contribution to the Anatomy and Systematic arrangement of the Cestoidea. Proc. Zool. Soc., London. Contribution to the Anatomy and Systematic arrangement of the Cestoidea. Proc. Zool. Soc., London. Contributions to the Anatomy and Systematic arrangement of the Cestoidea. IV. Ona species of Jnermicapsifer from the Hyrax, and on the genera Zschokkeella, Thysano- Proc. Zool. Soc., London. Mecorrt, F. J. (1921). Ona New Cestode from the Pouched Rat, Cricetomys gambianum. Parasitology, Z, guineensis* Varies from the ant to about the middle, the same strobila. May be in two sepa groups or these may united by a bridge varying numbers testes, in the si strobila. pouch. glands posterior. As in Z. muricola From 3 to about 20. Note—Apparently egg capsules first tain about 20 e, 457 MOSQUITOES AND OTHER BLOOD- -SUCKING ARTHROPODS OF THE UPPER SHIRI RIVER, NYASALAND BY . DR, J.B. DAVEY AND PROFESSOR R. NEWSTEAD (Received for publication 23 November, 1921) This paper deals with the mosquitoes and other sanguivorous Arthropods observed in a relatively small area on the banks of the Shiri River, a little south of Lake Malombe, in the Nyasaland Protectorate, British Central Africa. Our captures were made during the dry season, from July to the beginning of November, in ° the year:1911. No special effort was made by us to collect such material, and no search for the breeding-places of mosquitoes was undertaken. Our camp occupied a position about 200 yards from the river, the intervening space being comparatively free from ‘bush’ or other forms of vegetation. NHereabouts the river banks were -low, and partly or wholly submerged during the rains. Nearby the banks were elevated and clothed with tall forest trees, and beneath them there was a dense and almost impenetrable undergrowth. Two-thirds of the river was rendered impassable by the ‘ sudd,’ which was composed of a dense floating platform of aquatic plants, serving as a retreat for many species of birds, the crocodile and the hippopotamus. The sudd consisted largely of grasses, and in places extensive colonies of papyrus; the fringe being composed chiefly of the beautiful ‘water caltrops’ (T7afa bispinosa) and the equally troublesome ‘calbsbage’ or ‘ duckweed’ (Pistia stratiotes). MOSQUITOES (CULICIDAE). Anopheles (Myzorhynchus) mauritianus, Grandpré. This fine Anopheline was by no means common in our camp, as in all only ten examples were captured ; nearly all of these came into our dining-hut 458 while we were sitting at the table after dark. It is just possible that they may have been attracted by the artificial light; but of this we could not be certain. It did not seek shelter in the tents, as in the case of Anopheles funestus, and, therefore, does not appear to be a strictly ‘domestic’ species. With one exception, the specimens were of the dark form such as has been recorded from south of the Zambesi, and were characterized by the absence of spots either upon the costa or the fringe; the palpi presented three very narrow bands, the tips were in most cases black. The dates of capture were :— July, 1911; 1st August, 1911; 3rd August, 1911; 16th September, 1911; 18th September, 1911; 21st September, IQII. Anopheles (Cellia) pharoensis, Theobald. One specimen only. This was taken in the dark-room tent, 3rd August, IQI1. Anopheles (Pyretophorus) costalis, Loew. Only one example of this mosquito was captured, during the month of August. Anopheles (Myzomyia) funestus, Giles. This malaria-carrying mosquito was the most abundant of all the Anophelines observed by us. On July 31st, we decided to fix our permanent camp on the banks of the Shiri, opposite Matutas village, but our tents became so badly infested with this mosquito that we decided to abandon the site, selecting a more open spot a short distance away. Our first camping ground in this locality was surrounded by tall and, for the most part, densely foliaged trees, and was distant from the river about 80 yards. The permanent site of the camp was 200 yards from the water in practically open country. We found, however, that this Anopheline was equally abundant in both places: on August 2nd, one hundred and eleven specimens of this species were counted in cne of the tents, and there were certainly an equal number present in the other tent. The temperature on this occasion at 3-a.m. was 46° F., at midday 86°F. in the shade, and at 8.30 p.m. 55°F. On August oth, one hundred and sixty-two specimens of this species were captured in one of the tents. These consisted of thirty Gd, twelve unfed 99 and one hundred and twenty 2 92 which contained blood. This total does not, however, represent the actual number present in the tent, as large numbers escaped and many remained uncaptured, so that there were probably. half as many again as the total captures, and there was certainly an equal number present in the other tent. This was the only occasion on 459 which we attempted to catch as many individuals as possible with the view of ascertaining the ratio of sexes, the proportion of fed and unfed females, and the approximate number present in the tent. Further captures were made on August 14th and 31st, when twenty-three 9 9 and three dd, sixty-eight 9 9 and eleven dd were captured respectively. We consider that the figures given for August 9th may be taken as representing the approximate numbers present daily in either tent at that period; but we noticed even larger numbers on several occasions. During this time the tents were opened daily at each end, and as many mosquitoes as possible driven out, but as already stated we found no diminution in the numbers. The largest numbers observed were always in the darker portions of the tent, such as the angles of the roof and sides, between the boxes and on various garments. We also observed that they settled freely upon freshly skinned birds, especially guinea-fowls. We also observed on many occasions large numbers flying into the tents in the early morning between dawn and sunrise, the chief point of entrance being at the upper portion of the opening, through which they passed in more or less continuous flight until the rising sun put an end to their movements. Our native employees, about fifty in number, slept in the open about 15 to 25 yards from our tents. The nearest native village being on the opposite side of the river and about half a mile distant, we came to the conclusion that the principal food supply of these mosquitoes was obtained from the natives in our camp. Mansonioides uniformis, Theobald. It was by far the most abundant of all the mosquitoes, and also the most vicious and persistent biter. It attacked one at any time of the day or night, but was most troublesome shortly after sunset. It simply swarmed along the river and its immediate vicinity, but occurred also in the bush in places far remote from water. It was not, however, ‘domestic’ in its habits, though a few specimens were taken in our tents during the day. It was equally obnoxious on the lower Shiri; but almost entirely disappeared in the Zambesi where the margin of the river was free from aquatic plants. It is highly probable, therefore, that it breeds chiefly in those portions of the Shiri where the ‘sudd’ is extensive, and where rootlets and stems of aquatic plants suitable for the attachment of the larvae abound. 460 - Culex tigripes, Grandpré. One female only was taken in one of the tents in our camp during the month of August. -Ingramia (Mimomyia) uniformis, Theobald. One female of éhis rather rare mosquito was taken at night in our camp on August 11th, i9t1.. The beautiful pale blue reflections.on the thorax were very marked in this example. ~ Etorleptiomyia mediolineata, Theobald. One female was takes at the camp. ‘Taeniorkynchus aurites, Theobald. Examples of this species were taken in the camp, the date for which is now lost. PSYCHODIDAE. Phlebotomus minutus var. africanus, Newst. Three examples of this species were captured during the daytime, while at rest inside the tents; and one at artificial light, at 7.30 p.m. Other specimens, presumably of the same species, were also seen inside the tents at various times generally resting upon the canvas roof. On two occasions late at night examples attempted to bite one of us while under the mosquito -net; the familiar high-pitched note, somewhat resembling that produced by a mosquito but much fainter, was heard distinctly as the insects hovered round the face of its would-be victim; on the first occasion we were sleeping under canvas; the second time in our large mud-house. The first example was taken in the middle of July; the others were seen during August, and the last one towards the end of September. It is evident, therefore, that this insect occurs in Nyasaland during the dry season as it does also in Malta, and possibly also in other parts of Africa. TABANIDAE. Tabanus taeniola var. variatus, Walker, occurred very sparingly towards the end of August, four specimens only being seen and captured. These were all females. One was captured inside one of our tents, one was caught on a dead hippopotamus, and two were attracted by the fresh, moist mud on the walls of our hut. Tabanus africanus, Gray. Two females of this handsome species were also attracted by the fresh mud which covered the walls of the hut. None was seen after the moisture had evaporated from the mud. 461 PUPIPARA. Echestypus sepiaceus, Speiser. A number of specimens were taken from a young Kudu bull which was killed five miles west of our camp on September 14th, IQII. Olfersia ardeae, Macq. Two examples of this Hippoboscid were taken from a freshly killed Goliath heron (Avdea goliath), August 22nd, 1911. It is an extremely active insect, and the specimens were caught with difficulty. HEMIPTERA (Family CAPSIDAE). Trigonotylus brevipes, Jak. This minute green bug was first observed in our camp an hour after sunset, on August 1oth, 1911, when it bit one of us severely on the back of the hand; the proboscis being driven so firmly into the skin as to prevent its immediate escape, so that one was enabled to examine it carefully with a pocket-lens. Subsequently this insect occurred in large numbers at irregular intervals during August and September, always apparently attracted by artificial light, and always annoying by its persistent efforts to bite. It cannot, however, be considered a true blood- sucking insect, and we believe that its bites are made out of mere curiosity rather than to obtain blood. In life, it is of an almost uniform grass-green colour, with a narrow, pale streak below the costa of the elytra, antennae dull crimson-red, basal portion of terminal segment with an indistinct greyish ring; eyes black; abdomen vivid green; legs slightly paler, especially the hind femora; terminal segments of tarsi black. TIcKs. IXODIDAE. Though a number of animals were examined during our stay in the Shiri Valley, comparatively few of them were found infested by ticks. The following is a list of all the species observed. Rhipicephalus neavei, Warburton. One male and four females ‘from a Kudu, five miles west of camp, IQII, many specimens from a buffalo, 3rd August, 1911; two males and two females off Nswala or Mpala antelope, Upper Shiri Valley, 8th August, 1911 ; one female off an eland, Upper Shiri Valley, 20th August, 1911. 462 Rhipicephalus falcatus, Neumann. Off buffalo, 3rd August, IQII. Rhipicephalus stmus, Neumann. Three males from buffalo, Upper Shiri Valley, 3rd August, 1g11. Rhipicephalus maculatus, Neumann. One male from buffalo, Upper Shiri Valley, 3rd August, 1911. Hyalomma aegyptium, Linn. Off buffalo, Upper Shiri Valley, 3rd August, IQII. 463 BREEDING PLACES OF ANOPHELINE MOSQUITOES IN FREETOWN, SIERRA LEONE BY B. BLACKLOCK (Received for publication 29 November, 1921) PLATES XXVII TO XXXI AND THREE Maps Stephens and Christophers (1900) stated ‘in Freetown we found that during the dry season the streams were the main source of Anopheles,’ Daniels (1901), in a letter to Sir Ronald Ross bearing on the anti-mosquito operations then proceeding in Freetown, expressed the opinion that in the dry season, while the surface collections of water would have disappeared, new breeding-places would arise, ‘mainly the streams, small and large, which remain, possibly some of the other wells and artificial collections of water in tubs, etc.’ Boyce, Evans and Clarke (1905) mention ‘the great disadvantage which attaches to the streams in the dry season, viz., that they. constitute in Freetown, at that period, the chief sources of Anopheles supply.’ With the object of ascertaining whether the streams referred to -above, which traverse Freetown, still act at the present day as breeding-places of anopheline mosquitoes, I took the opportunity of examining them at the end of the dry season of 1921. The search for larvae was carried out in the month of May, at which time the stream beds contained comparatively small amounts of water. The plan adopted in making the survey was to start at the mouth of the stream at the point of entry into the sea and to work up stream through the town until the places of origin of the streams was reached in the high ground at the foot of the hills behind the town; the various tributaries were followed in turn. The results obtained showed that in each stream in which larvae ' were found, breeding-places were present in largest numbers at the 464 lower end of the stream; as one proceeded upwards through the town the breeding-places and the numbers found became scanty, while on emerging from above the town breeding places were again found, but still in small numbers in comparison with those at the lower end of the streams. The reasons for this distribution are doubtless numerous, but some of them are clearly connected with the conformation of the ground. The streams, in some cases, by the time they reach a point several hundred yards from the sea have produced wide, steep-sided gorges through which they pursue a tortuous, irregular course; the numerous sheltered bends, overgrown with grass and weed, afford excellent breeding-grounds and shelter for the larvae. The process of erosion in the wet season appears to have more than compensated for the additional volume of water to be carried, so that the advantage of flushing is to a certain extent lost in the lower portions of the streams. In the town proper the streams are passing through a rocky formation, with the result that there is slower erosion; this and the great amount of canalisation of tributaries which has been carried out in recent years permit of more complete washing out of the stream bed in its passage through the town. Above the town the prevalence of larval breeding-places was to be anticipated from the more diffuse nature of the water courses, their extensive area, and the plentiful vegetation through which the water slowly percolates before forming definite streams. The conformation of the ground, while it favours the presence of Anopheles by providing suitable breeding-places, acts also in their favour, especially in the gorges mentioned above, by rendering it difficult to obtain access to these breeding-places for the purpose of detecting them, and still more by rendering the application of the measures necessary for their eradica- tion very laborious. The maps appended show (Nos. I and IJ) the places in which anopheline larvae were discovered and the manner of their distribu- tion in 1921 and 1900. It was observed that the residual breeding- places in the streams at the end of the dry season are of two kinds. The first is the edge of the winding and eroded bed of the stream just before its entrance to the sea, the second is the shallow water, well protected by vegetation and extending over a large surface, which is found at the places of origin of the streams. These residual Annals Trop. Med. & Parasitol., Vol. XV L PLATE XX] Puorocrapu No. I Anopheline breeding place, in the town. The man is standing in the water at the point in which A. costalis larvae were found. Note native house in the background, water sluggish. 465 Seale mile | FRE E TOWN soe m Non WV aii fy Ui, Mar 1. @ A. costalis larvae or pupae found 1921. mile ! FREETOWN - —_— Mig Y i, Va 2 il Coins ln, Me 5 = ily pri Anopheles Map 2. Anopheles larvae found 1900. 466, breeding-places doubtless contribute very materially to the spread of Anopheles when the rains commence, and other breeding sites become available. Some of the more interesting breeding-places found are seen in photographs 1 to 5. SPECIES OF ANOPHELES FOUND Larvae and pupae collected from the various breeding-places were taken to the laboratory and allowed to develop ; anophelines which emerged belonged in all cases to the species A. costalis. This appears at the present time to be by far the commonest anopheline breeding at the end of the dry season in Freetown. It is deserving of note that in the early days of mosquito investigation in Freetown this species predominated at the end of the wet season also. Daniels (1901) mentions Anopheles costalis as a common mosquito in Freetown, and further states that ‘A. fumestus was found near but not in Freetown.’ Ross, Annett and Austin (1902) stated that while of over two hundred anophelines obtained from Wilberforce Barracks all were A. coséalis, they obtained from Dr. Berkeley at Kissy both this species and also A. fumestus, the latter ‘by far the more numerous.’ They add that A. fumestus was restricted entirely to the eastern part of the town, and that they never found a larva or an adult of this species west of Government House. It is not stated, however, how near to this dividing line A. funestus existed on the east side at that time. Thus, while there is no evidence from the foregoing and the present survey that the proportion of A. funestus to A. costalis has undergone a change during this period of years, there is considerable evidence to show that A. costalis was then, and is to-day, the commonest anopheline in Freetown. Owing to lack of time, little could be done in the way of dissections of adults, either caught wild or experimentally fed on malaria carriers; this will be reserved for a further date, when it will be of great interest to discover whether there exist to-day .any rates of mosquito infection comparable with the Wilberforce Barracks figures of Ross and collaborators in 1902, namely, twenty-seven infected Axopheles costalis in one hundred and nine captured females dissected. Annals Trop. Med. & Parasitol., Vol. XV “\PLATE XXVIII Puotocrapu No. 2 Anopheline breeding place, above the town. The water is sluggish and almost concealed by vegetation. Numerous 4. costalis larvae found to left of man. Sous 467 lt may be said then that the results obtained in 1921 at the end of the dry season were such as to confirm the predictions of twenty years ago, and in view of the great amount of careful and pains- taking work which has been steadily carried out in the interval with a view to abating mosquito breeding, it is clear that in these streams we have a problem which will require a little more detailed investigation. METHODS OF DEALING WITH THE STREAMS Previous suggestions as to how such an obvious source of Anopheles could be dealt with have been of various kinds. Some of these may be- mentioned. Daniels (1903) wrote: ‘Two possible methods which are most obvious are the formation of a central channel in the bed of the stream, with larger. collections of water in sufficient numbers of places for drinking purposes, and, lower down the stream, other places for washing, etc. The second, which might be cheaper but certainly less effective, would be to dam up the streams so as to obtain a sufficient head of water to flush out the whole channel at intervals.’ Boyce, Evans and Clarke (1905) summarised their views on this matter as follows :— (1) Reconstruction of the bed of streams. (2) Diversion of water for flushing of town drains in dry weather. (3) Construction of dams in streams and flushing of beds at intervals. It will be seen that schemes one and three here correspond to the two suggested by Daniels above. The town drains referred to, in number two, are the surface drains in the streets. (1) Reconstruction of the bed of streams. If one follows such a stream as Nicol’s Brook from its origin above Foulah town to the point where it enters the sea, it is realised that the operations involved in this scheme would be of great magnitude. The process of erosion has been so irregular in its action that the - water does not flow on a simple slope from above down to the sea 468 level. It forms at places considerable falls, and also has cut very large cavities in the bottom of the stream bed. Laterally also it extends in different places to very different dimensions. In order to rectify the levels and build a channel capable of carrying all the rainy season water, it appears that a very large expenditure would be involved. If when this expenditure had been made the streams could then be regarded as safe, it might be a plan worthy of consideration: but it appears probable that constant attention would be required to ensure that the lateral tributaries discharged properly into the central channel and that pools did not form outside this channel itself. This would involve one of two things. Either the channel would have to be so levelled and sloped at the sides that no pools could possibly form—a vast operation—or else such pools would have to be treated regularly with larvicide, as at present. The other streams would require similar measures. (2) Diversion of water for flushing of town drains in dry weather. This scheme would, it appears to me, fail to be effective in preventing anopheline breeding in the stream beds, for two reasons. One is that water oozing out of springs between the strata in the bed of the stream would still form pools suitable for breeding anophelines.. The other is that many of the drains referred to discharge into these streams and would thus carry the water back to the bed of the stream after having flushed out the town street drains. (3) Construction of dams in streams and flushing of beds at intervals. This scheme assumes that the bed of the stream is capable of being effectually flushed, but the observations which were made above in connection with the conformation of the streams go to prove that Nature has already by the process of excavation and erosion effectually prevented such a flushing action being successful. Further, it cannot be said that even a large volume of water passing occasionally down a river bed which is a honeycomb of pools will ensure the sweeping out of anopheline larvae from these pools. This scheme, therefore, does not appear to guarantee success. Annals Trop. Med, & Parasitol., Vol. XV PLATE XX1X Puorocrapn No. 3 Anopheline breeding place, in stream bed in the town. A. costalis \aryae found near bank to left of where the woman is standing. C. Tinling & Co., Ltd., Imp. 469 SEGREGATION METHOD '- Stephens. and Christophers were among the pioneers of the segregation method of dealing with the malaria problem, and there is ample evidence of the.efficacy of this method where it is vigorously and thoroughly carried out. It is chiefly of value where we. are dealing with a small non-immune community which has to exist side by side with an overwhelming preponderance of immunes and semi- immunes. It is based on the fact that there always has been, and always is, a large proportion of infected children among the native population. If we believe such conditions will always remain in the future, it is clear that the readiest and most inexpensive way of dealing with such a small white community will be by strict segregation; in this way a great degree of safety from infection can ' reasonably be insured. In places which are being newly developed this method can be used advantageously, as also in places where the white population is small and circumstances favour their settle- ment away from the native. But in old established towns it is not by any means an easy method to apply, more especially where the white population is growing, and likely to grow greater year by year. It is also a method which leaves the natives to themselves and frankly regards them as so infected that no immediate and inexpensive mode of dealing with this infection is available; to this extent it is a method which is a tacit acknowledgment of defeat. Segregation occupies in respect to the malaria problem much the same place from the point of view of medical prophylaxis as evacuation of fly areas occupies in respect to the trypanosomiasis problem. Each may be excellent as a temporary expedient, but neither can be regarded as a final and satisfactory method of dealing with these problems. The bearing of segregation on the question of how to deal with the streams of Freetown is clear when we consider the results obtained by Stephens and Christophers in their investigation on the anopheline content of these streams and the native houses situated close to them. I have obtained the permission of these authors to reproduce a spot plan (Map III) which they made in 1900, showing the distribution of adult anopheles in houses near such a stream. It will be cbserved that ‘houses in which anopheles are present in enormous numbers’ are found immediately beside the stream, that 470 ‘houses in which search in early morning reveals a few anopheles’ extend back from the stream a short distance, while as we recede further from the stream we reach ‘houses in which anopheles cannot be found.’ The conditions depicted here would, I feel no hesitation in saying, reproduce themselves automatically immediately any considerable relaxation of the present sustained effort on the part <8 Houses in which anopheles are present ‘in enormous numbers B Houses inwhich search in early morning reveals afew anopheles 1 Houses inwhieh anopheles cannol be found Map III. of the sanitary authorities of Freetown took place. In Freetown it does not appear to me practicable now to adopt a method of rigid segregation, and moreover, even if it were practicable, it would still be advisable to deal with the problem of the streams with a view to ameliorating the condition of the native. Annals Trop. Med. & Parasitol., Vol. XV PLATE XXX Puotocrapn No, 4 Anopheline breeding place, in the town. A few A. costalis larvae were found at points along the edge from where the man is standing; water fairly rapid. C. Tinling S Co., Ltd., Imp. 471 IS A PERMANENTLY EFFECTIVE SCHEME POSSIBLE? The only permanent scheme which will be effective in dealing with the stream beds appears to be one which obliterates them entirely. Whether any such scheme can be devised is a matter for engineers to determine. It might involve the construction of a canal or canals capable of carrying the water to east and west from above the town and reclamation of the present stream beds. The fact that the outlay required, if the scheme were practicable, would be very large indeed dces not enter at present into the discussion. Whether the effective method will prove too costly need and can only be decided after considerable investigation over a period as to whether an effective scheme is practicable from the engineering point of view. It deserves investigation, however, even if apparently excluded on account of prohibitive cost, for we are not justified in assuming that an effective method which would be too expensive at present must necessarily prove so in future years. REFERENCES Boyce, R. Evans, A., and Crarxg, H. H. (1905). Report on Sanitation in Freetown. Liv. Sch. Trop. Med. Memoir. Vol. XIV, pp. 32 and 33. Daniets, C. W. (1gor). Liv. Sch. Trop. Med. Memoir V. Pt. 1. Appendix, p. 15. Ross, R. Annett, H. E., and Austen, E. E. (1902). Report of the Malaria Expedition. Liv. Sch. Trop. Med. Memoir II, p. to. Stepuens, J. W. W., and Curisropuers, S. R. (1900). ‘The Native as Agent in the Malarial Infection of Europeans.’ Reports to the Malaria Committee of the Royal Society, p. 6. ae ‘areezoa Swuuoe AVITISSS ¥iS FHA Moe ia) Gre tee i x t iz bhi if tiw i ‘ +e a3 2 i) svads-mot Mow Bue teks So sspeeraee a feted th) vebor eiende tooresg. ot ta. nO . te S20 4RRT IS 1 ord end Bea ‘ tence Relea i oft «3 iat ae ¥ sf} * fom f “+t agjtoolis od [fed rlonlw Sides Irorgarmed . 5 eylgrolii ‘sw ano ad of emaqas AbSdi : oi borivib al seo-smedor done outaana o@2 svlovar migiat a prow i feaTiaa wht ayy? agraioe lt EY a ceeih atid ehak seseste Fe Ie i am ute Dae Beart ‘eliaad bor arog: Nii oy ot ae bore al pre adiisgriearnt ae fer} area uid tie Sul? iaotald festa say ‘a 1 £9 herve: ii as >... 3945 qh Jove reer om 6 am tote are nur ye}, daca > da svivasqes Go) a7) AIRGwe Hide I amn'e cnet ae re hor cE ae ve ny) y wien 3 ct TS Sisesith co A A r® eitlelh ira, -.tRoga) A hot baat cm: ; re ive ihre + Yeast, Va wd atk Qa a weap . " - * = ; - 4 ; ie 7 wee, an, oa Annals Trop. Med. & Parasitol., Vol. XV PEATE SOXCEE PuotoGraPH No. 5 Anopheline breeding place, in the town. Behind the rock in front of which the man is standing is the pool in which little vegetation was present ; a few A. costalts larvae. C. Tinling FS Co., Ltd., Imp. 473 NOTES ON AN APPARATUS FOR THE IN- DIVIDUAL BREEDING OF MOSQUITOES BY B. BLACKLOCK (Received for publication 29 November, 1921) The apparatus here described is designed to facilitate the task of breeding out mosquitoes individually from any stage in which they happen to be at the time when they are collected, and in this way obtaining a record of the association of the various stages. It is a development of the breeding apparatus which Carter and Blacklock (1920) described, and its small size makes it easily portable and convenient for use in the tropics. By means of it, with a little trouble, a complete record of the stage originally obtained, the adult stage, and the intervening stages is possible. The parts of which the apparatus consists are glass capsules of suitable size, embedded in a wooden base, glass cylinders of which one end is covered with mosquito netting, and glass tubes for the collection of the casts at the various stages of development. It will be simplest to give the dimensions of the model actually in use; this has proved, in so far as mosquitoes are concerned, satisfactory, the mortality, whether of larvae, pupae or adults, being comparatively small. Modifications could easily be introduced which would render the apparatus suitable for observing the development of other insects in their aquatic stages. The capsules. These are made from glass tubing, and are of the following dimensions :—Internal diameter, 11 mm. ; internal height, 18mm. They are of stout glass, about 1 mm. thick, rounded at the bottom like a test tube (fig. 1), and they fit into holes drilled in the Fic. 1. wooden base at measured intervals. The depth of the hole bored is such that when a capsule is placed in it the edge of the capsule lies 474 flush with the surface of the board, and the width of the -holes 1s. such that it is possible with fine bladed forceps to lift the capsule out of the board. A small ring of plasticine can be used to fit the junction of the holes that are too large for the capsules. The board. The wood used in making the board is hard and well seasoned so that it will not warp. The length is 30 cm., the width 11 cm., the depth 3 cm.; the holes are bored in three rows, the distance between the holes from each other in any row or between the rows being 2 cm. (fig. 2). In this manner it is possible to have in each board twenty-seven capsules arranged in three rows of nine capsules each. In boring the holes, the first row is bored near the edge of the wood, leaving a rim of only about o'5 cm. in front of them; the object of this arrangement will be explained later when describing the manner of using the apparatus. The cylinders. These are made of glass, of similar thickness to that used for the capsules. The dimensions are :—Internal diameter, 15mm.; height, 25mm. It is essential that one end at least of the cylinder should be so cut that it will stand upright and make fair contact all round its edge with the board on which it stands, if one end is irregular, it should be used as the upper end; this end is in all cases covered with a small piece of mosquito netting, kept in position by a piece of string or two thin rubber bands (fig. 3). 475 As rubber is apt to deteriorate in the tropics and a band may break, it is necessary when using such bands to use two to ensure safety ; string, however, answers the purpose quite satisfactorily, merely taking a little more time to fix on and take off. Fig. 4 gives a section of the cylinders in position covering the capsule embedded in the board. Tubes for casts. These are made from glass tubing of an internal diameter of not less than 5 mm. and about 6 cm. in length, drawn to a blunt point at one end (fig. 5). When the larval and pupal Fic. 5. pelts have been placed in them they can be corked and waxed, or better, if so long as to permit of it, they can be drawn out in a small flame and sealed. Method of use. The stages collected in the field, and from which it is desired to breed adults, are placed in a dish of such a size as to render it easy to ensure the isolation of individuals, and one by one these are transferred to the capsules, each into one capsule; this is filled with water and replaced in its position in the board, as evaporation occurs the fluid lost must be replaced daily to keep the water at the surface level. This is especially necessary in the case of anopheline larvae. The capsule is marked by a number written 476 on the board in front of it in pencil, the same number being entered in the book in serial order. The front of the board is that margin on which the narrower rim is left; this serves to identify the front, and is useful in practice also by leaving standing room at the back of each capsule for the accommodation of the cylinders when they are removed from their capsules. The capsule having been filled with water and replaced, a small portion of dried, powdered cockroach, or other suitable food material, is deposited on the surface of the water with a needle. Each day the capsule is examined at intervals, and when a cast skin is observed it is picked out with a needle or fine camel-hair brush and transferred to one of the tubes which have been described above, this being filled half full of preserving fluid, or the whole contents of the tube, including the larva, may be pipetted out with a wide bore pipette into a small porcelain dish and the cast skin recovered, the larva being replaced. Transfer by means of a pipette is apt to lead to difficulties owing to the fact that whereas the larval pelts usually fall to the bottom, the cast pupal skins usually float and may become stranded on the side of the pipette used. The date of the stage is noted in the record, the tube is numbered with the same number as the capsule and is retained for all material which is derived from that capsule. As soon as the pupal stage is reached one of the cylinders is placed in position over the capsule, and when the adult emerges into the cylinder this may be pushed carefully back off the capsule on to the board immediately behind it, in order to give the adult time to dry and expand. At the same time the pupal cast is removed and placed in its tube, which is then corked and waxed or sealed over a flame. When the adult is ready for pinning it is removed from the board by slipping a strip of thin, stiff paper underneath the cylinder containing it, chloroformed by sliding the cylinder on to a piece of paper moistened with chloroform, while the upper end of the cylinder is covered by the finger; when pinned it is labelled in the collecting box with the same number as the capsule from which it was derived and the tube containing its other stages. Before use again after one experiment the capsule is carefully swabbed out with a pledget of cotton wool, first with water, then with alcohol, and dried. In this way it is possible to obtain in the majority of cases a very complete record of the stages from the time the individual was 477 collected ; the apparatus is of value, not only as a simple means of associating stages and noting their duration in the laboratory, but also for isolating mosquitoes as they emerge if required for experi- mental purposes. Precautions in use. Care is required with reference to the following points :— (1) Proper entry in the record of the number given to the capsule, pelt-tube and adult. (2) That one individual only is introduced into the capsule; this is especially necessary when dealing with young larvae or eggs. (3) That each pelt is removed as soon as it is shed. (4) That the tube containing pelts is well sealed if intended to be kept for future examination. ae h yw , used Ohare Oa vied: Tae oN Ue. i ‘glothgge & SN i his al adit: Neowh ete ‘ide pth Se ct Dt a 4 i or a ok ie ’ at «% A Pte 1: er i} Asi 45080 tt ttt ah 45 3 A Pac? Been Yervaxyt be oy ¢ wert "ay | ae hey $4 ae ' : ‘Huha Ban fy tes “lis ee ; : wont : «iy Oa ES . Es aheans 4 Gint Bas fete ee whit: pet ty ieit it i ns. Shc if } ot t's ri] aly aed 1 * aie tis ee, suri ae ; : ¥ a rf ; 2 ior) oe bvOuts = if, 3 8h. tyatree “sy ab sy eetea ING Oe 3 Dee he Mea nt EF _ — ™“ <. ~ 5 , . “ ney J Pe P ‘iste m ig ett bee oa “oo ris v coun) =h. dai} & x veg’, 4 f 4 ‘as ee j i a 5 ae ¥ Wh aS a x ie (nape . i ' , ln deal ' in ae o = “ Suey he ry Se . ES g wed fsind it ore. vse ou te al, jo hs ye gaa — oe | od hak). i Lp ae . r ‘ - , j ’ - . 7 p 7 - nt f . ¥ a ae 479 THE TREATMENT OF A CASE OF RHODESIAN SLEEPING SICKNESS BY THE PREPARATION KNOWN AS ‘BAYER 205’ BY WARRINGTON YORKE (Received for publication 6 December, 1921) The patient (G. J.) contracted the disease in the Seringe district of North Eastern Rhodesia on the Congo Zambesi Watershed. He was suddenly taken ill on 17th September, 1920, with severe headache, pains in the back and neck, great prostration and a temperature of 105° F.; during the first eight days of the illness the temperature varied between 104°F. and 106°F. Trypanosomes were found in the blood on 27th September, and the patient was thereupon sent to Broken Hill, where he arrived on 24th October, and was treated by Dr. Wallace with tartar emetic, intravenous injections (2°5 to 3 grains) being given every other day. A little later intramuscular injections: of antimony oxide (4, grain) and soamin (2 grains) were also given. Between 24th October and 26th December 3°28 gm. of tartaric emetic, o'1 gm. of antimony oxide and 2°47 gm. of soamin had been administered, with only a moderate degree of benefit to the patient: trypanosomes were frequently found in the blood and there were several severe febrile disturbances. Between 26th December and 15th January, 1921, thirteen injections of stibenyl (in all 3°05 gm.) were given. These injections were, however, badly borne, and as trypanosomes were present in the blood on 16th January, the original treatment, with tartar emetic, antimony oxide and soamin, was resumed. Things improved somewhat, and on the 3rd February the patient was well enough to leave for England. At Cape Town, and on the voyage, he had further injections of tartar emetic. When the patient reached Liverpool on 12th March, he was in fairly good condition; trypanosomes were not present in his blood 480 (inoculated rats did not become infected), there was well-marked - autoagglutination of the erythrocytes, and the weight was 144 lbs. Daily examinations of the blood were negative until 18th March, when a few trypanosomes were found. Inoculation of rats showed the parasite to be 7. rhodesiense. Intravenous injections of tartar emetic in doses of from 2 to 3 grains were then given on alternate days. Neither these nor injections of atoxyl, however, seemed to exert any influence on the course of the disease. Violent febrile disturbances accompanied by severe headaches and great prostration, and the appearance of trypanosomes in the peripheral blood in considerable numbers, occurred with the utmost regularity every five or six days, and the patient went steadily down hill. In view of the fact that the infection appeared to be antimony and arsenic resistant, I endeavoured to obtain from Professor Mayer, of the Hamburg School of Tropical Medicine, a small quantity of a preparation called ‘ Bayer 205’, manufactured by the firm of Friedr. Bayer & Co., Elberfeld. Although this drug—the constitution of which was not disclosed on account of the state of the Germany chemical industry—had not been used in any case of human trypanosomiasis, it had been employed with remarkable results in the treatment of experimental trypanosomiasis of animals, records of which had been recently published by Haendel and Joetten (1920) and by Mayer and Zeiss (1920). Professor Mayer informed me that he was unable to comply with my request to let me have a supply of the drug for trial, but stated that if I sent the patient to Germany they would be glad to treat him at the Hamburg School. This, unfortunately, resulted in some loss of time, as I did not like the idea of sending the patient to Germany to be treated with a proprietory article, the composition of which was not disclosed and which could not be sent out of the country. However, as notwithstanding the most rigorous treatment the patient’s condition got steadily worse, I went critically through the above mentioned papers again and was so impressed by the results recorded, that I overcame my own scruples and those of the patient, with the result that he left for Hamburg on 5th July, 1921. During his eight and a half months illness the patient had received, in addition to the other drugs mentioned, 22°7 gm. of tartar emetic, and at the time he left for Hamburg his weight had fallen to 481 132 lbs., he was suffering from frequent violent febrile disturbances, antimony and arsenic preparations were without any effect on the course of the infection, and the condition seemed almost hopeless. The following details of the case are taken from a recent paper by Miihlens and Menk (1921), under whose care the patient was when in Hamburg. On oth July, two days after his arrival, he had a rigor, accompanied by headache and fever, and trypanosomes were found in the blood. Treatment was immediately commenced, ‘Bayer 205’ 05 gm. in 5 per cent. solution being given intra- venously. The injection was well tolerated; the fever subsided in six hours instead of in the customary thirty-six hours, and trypano- somes had disappeared from the blood within sixteen hours. The next day a second injection of 1 gm. in 10 per cent. solution was given, and this was repeated on the following day: both injections were well borne and the urine remained free from albumen or casts. The blood was examined twice daily during the following seven days, with negative results, and a.further injection of 1 gm. was then given. As a small quantity of albumen and occasional red and white blood corpuscles appeared in the urine, no further treat- ment was administered. The patient improved remarkably, rapidly regained his sense of well-being and put on weight. Almost daily examinations of the blood were made, until 29th August, but with negative results: there were no further febrile attacks or any other untoward symptoms, and on 13th September the patient returned to Liverpool. When I saw him on 15th September, I was greatly impressed with - the remarkable improvement in his general condition: he had lost the anxious and weary expression which had been so characteristic for some menths before he went to Germany, his weight had increased to 145 lbs., the blood was negative and sub-inoculated rats did not become infected, and the autoagglutination of the erythrocytes had disappeared. The urine contained a trace of albumen, otherwise the patient seemed perfectly well. The following day he left Liverpool for his home in South Wales. On 5th November, 1921, he returned to Liverpool for further examination. He appeared in excellent health, and stated that he never felt better in his life and had played two rounds of golf daily without the slightest distress. The blood was examined microscopically and also inoculated into three rats, 482 with negative results: there was no autoagglutination of the erythrocytes or any other sign of the disease: the weight had increased to 150 lbs. (patient’s normal weight), the urine, however, still contained traces of albumen. On 11th November he sailed for South Africa. The above record is of peculiar interest, as it relates to the first case of human trypanosomiasis treated with adequate doses* of ‘Bayer 205’. Whilst recognising that hasty and premature claims of success in the treatment of a disease like sleepmg sickness are to be deprecated, and that the further history of the patient alone can decide whether a real therapia s¢evzlasans magna has been effected, the facts that the disease was of at least eight and a half months standing at the time of treatment with ‘ Bayer 205’; that the patient was extremely ill and had been getting steadily worse for months, that the infection was completely resistant to the ordinary prepara- tions of antimony and arsenic; that since the administration of ‘Bayer 205’ all symptoms of the disease have disappeared, the general condition of the patient has improved enormously, his weight has increased by 18 Ibs., his blood has been free from parasties and non infective to rats for an observation period of four months—these facts, considered in the light of the usual history of cases suffering from T. rhodesiense, warrant the hope that in ‘ Bayer 205’ we have a drug of exceptional trypanocidal power. REFERENCES HaenpbeEt, and Jorrren (1920). Berlin Klin. Woch., Vol. LVII, p. 821. Mayer, and Zetss (1920). Arch. f. Schiffs- u. Trop.-Hyg., Vol. XXIV, p. 257. MUutens, and Menx (1921). Miinch. Med. Woch., Vol. LXVIII, No. 46, p. 1488. * Miblens and Menk (loc. cit.) give details of the administration of the drug to a case of gambiense infection in March, 1920, but only two small doses (0"2 gm.) were given at an interval of a week. A relapse occurred and the patient was then treated with other drugs. ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY THE UNIVERSITY OF LIVERPOOL ANNALS TROPICAL MEDICINE AND PARASITOLOGY ISSUED BY THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE Edited by Proressor J. W. W. STEPHENS, M.D.Cantab., F.R.S. Prorrssor R. NEWSTEAD, M..Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S. Proressor WARRINGTON YORKE, M.D. Proressor B. BLACKLOCK, M.D. VOLUME XVI (March 31, 1922, to December 30, 1922) With Frontispiece, eighteen plates, one hundred and thirty-one jigures in text, and nine charts LIVERPOOL : THE UNIVERSITY PRESS OF LIVERPOOL LIMITED LONDON: HODDER AND STOUGHTON LIMITED eri. TUL OST oe ae VTA t pf of = 2 + ft (1 oe - ayaa =a ; Fel » , ; Sy woe } OTRYMITRAYW LAMA A © :t 9 re a 5 ¥ 2 TOCIMAYIT AHO 7 AEOIONS clea me CONTENTS No. I. March 31, 1922 Zammit, T. Undulant Fever in the Goat in Malta STEPHENS, J. W. W. Undulant Fever in the Naval, Military and Civilian Populations of Malta Moopy, L. M. Alastrim ; or, Kaffir Milk Pox Newsteap, R. A New Species of Phlebotomus from ‘Trinidad Newsteap, R.; and Evans, ALwen M. A New Tsetse-Fly from the South Cameroons Maptestong, P. A. Notes on Australian Cestodes. Part III ... Maptrstong, P. A.; and Souruwett, T. Notes on Australian Cestodes. Part IV Waterston, JAMEs. A Contribution to the Knowledge of the Bionomics of Sand-Flies Young, C. J. Human Intestinal Protozoa in Amazonas ... Bracktock, B.; and Apter, S A Parasite Resembling Plasmodium falciparum in a Chimpanzee PAGE 21 47 51 55 61 69 93 99 CONTENTS No. 2. July 20, 1922 Bracktock, B. PAGE The Signs of Filarial Disease 290 =e ane we a0 ae Bo he7/ Incram, A.; and Macrig, J. W. S. Two Further Cases of Cardiac Aneurysm ... ake 00 Se So sos, EG SouTHWELL, T. Cestodes in the Collection of the Indian Museum aoe ses 503 Bee 1 37/ Maptrstong, P. A. Cryptocotyle lingua (Creplin, 1825), Fischoeder, 1903, in a Dogin England ... 153 Macriz, J. W. S.; and Incram, A. On the Genital Armature of the Female Mosquito ote wes S00 coh CYHESF, Mapusstone, P. A.; and Sournwett, T. Notes on Australian Cestodes. Part V ... sce aes ze abe esa) AESG StepHens, J. W. W. The Incidence of a Disease in Population Groups, the Number of People in which is known or unknown ... A Ae a2 33 ws sce) eekQG Cawston, F. G. The Experimental Infestation of Physopsis africana we ses 200 are 27/ Evans, Atwen M. Notes on Culicidae in Venezuela, with Descriptions of New Species. Part II 213 Gorvon, R. M. Ancylostomes recorded from Sixty-seven Post-mortems performed in Amazonas 223 CONTENTS No. 3. October 18, 1922 Gorpvon, R. M. PAGE The Susceptibility of the Individual to the Bites of Stegomyia calopus ... or 220) ArcuiBaLp, R. G. Tuberculosis in the Sudan, with Notes on a Case of Breast Tuberculosis in a Sudanese ... ee Sor oe ae éch im ee re os MBS235 Incram, A.; and Macrir, J. W. S. West African Ceratopogoninae. Part II ... es = See see ceo 243 Bracktock, B.; and Apter, S. The Pathological Effects Produced by Strongyloides in a Chimpanzee ... a e258 Bracxtock, B.; and Apter, S. Pulmonary Lesions in Dogs and Cats Naturally Infected with Nematodes .... 291 Apter, S. Ancylostomes in Animals in Freetown _... in oa wie sis att 6203 Gorpon, R. M. The Occurrence of Ancylostomes Resembling Necator americanus amongst Domestic Pigs in Amazonas... a Ae sig san Fc me) 205 Gorpon, R. M.; and Young, C. J. Parasites in Dogs and Cats in Amazonas ... We sa wee “ae ase 2297, Incram, A.; and Macrie, Jj. W.S. A Note on the Prevalence of Ceratopogonine Midges on the Windows of the Accra Laboratory during a Completed Year oe ae om soa AFOUL Maptestong, P. A. Notes on Australian Cestodes. Part VI ... fas = op bon fre 305 Macriz, J. W. S. The Ascaris of Cattle ae va 55 po ies a Fes eee Gorpon, R. M.; and Evans, A. M. Mosquitoes Collected in the Manios Region of the Amazon _ ... ack SG ArcuiBazp, R. G. Trypanosoma rhodesiense in a Case of Sleeping Sickness from the Sudan 4h (339 ( vii | CONTENTS No. 4. December 30, 1922 ArcHIBALD, R. G. An Unusual Type of Nodular Leprosy in the Sudan Crayton LANE. Ancylostoma brazsliense Avtrr, S.; and Crark, E. J. Intra-uterine Infection with Ancylostoma caninum in Dogs SourHwett, T. Cestodes from Indian Birds with a Note on Ligula intestinalis... STEPHENS, J. W. W. A New Malaria Parasite of Man Youne, C. J. Notes on the Bionomics of Stegomyia calopus, Meigen, in Brazil Corson, J. F. The Occurrence of the Larvae of Onchocerca volvulus (Leuckart), 1893, in the Skin of Natives of the Gold Coast Srepuens, J. W. W.; and Yorke, WARRINGTON. A Case of Sleeping Sickness (I. gambiense) Treated by ‘ Bayer 205 ’ Gorvon, R. M. Notes on the Bionomics of Stegomyia calopus, Meigen, in Brazil, Part II Macrig, J. W. S. Observations on the Role of Cockroaches in Disease Evans, Atwen M, The Occurrence of Xenopsylla astia, Roths,. in West Africa Patrick, ADAM. Notes on a Case of Blackwater Fever Mackenzie, A. J. Case of Trypanosomiasis Macrig, J. W. S.; and Corson, J. F. Observations on Onchocerca volvulus Macriz, J. W. S.; and Corson, J. F. A New Species of Filarial Larva found in the Skin of Natives in the Gold Coast viii ) pr 4 PAGE 341 347 353 355 383 389 407 42 I 425 441 449 451 457 459 465 4 Pr Ayrice i ; H ae gt j 7 pe - 8 > of ‘ F ot esrecrs, Seaten coy Vaio tee oe ——SRDES OF pil THORS Aart a a, nh a “hy Pal ' ; J H i su ‘ é ‘ 1S, 0%. St << 7 wale i { - ‘ : - : " e ; Pe Ciy 0 FS ) wm Mie a | or 5 5, De ' % } 5 Ma. ay 2 i . ost. on? .é +4 GERTRAL 'KDZa Bk ; Z : err i ee ..c, aon - ar le tage gg «fi -_- er on | ee ee Po fst “alge ic ‘Jie @ hi senate a ied: tary eens LAN 4, a ee DP arrinesti . a akon th wes: seh Sear i? Nid, &, ae Goh. Ff, tar my oteriee fei wok 4, eemre iiies o58 51 # 9 125 (We Mairi ir, mee TDS ; bei Inpex or AuTHors GENERAL INDEX INDEX OF AUTHORS PAGE COTES QR a ae 293 Adler, S. ; and Blacklock, B ...99, 283, 291 Padlerine sand Clark, Wet) .2.cecs pseudo-punctipennis ......... 214 5 ceylanicum, alleged dis- 5 punctimacula ........seeeerene 216 tinction from A. 95 rufipes, female genitalia...... 163 braziliense ......... 225, 347 » Squamosus, female genitalia 163 s ceylanicum from cats and 3 venezuelae, sp.n., morph- dogs in Africa 227, 292, 293 ClO gy aeeacaes Wace oaeee eee 214 iv PAGE Anoplocephala perfoliata .......0.0..0000++ 193 _ DUI AT Sims from amphibians.............. 152 Us », the black swan........... 189 a Oli GUOWS Wd -ceseeee's maps ucsae 146 x Jp HAA GES 3 wots sesbaaetanes 145 = ae Undiay: birds: ascends. 355 os dedi pammals}* Lies wevees. os 127 5 Rn PIGEONS mcetaneezsenties = 136 BS Br PEE PUUES ipeeasaakiowtee ss 150 PAGE Chilomastix mesnili in Amazonas and elsewhere! “ES bea atc antomeeseces 94; 97 Chimpanzee infected with parasite resembling P. falci- PATUMIN Bacietn os eeeee es 99 5 infected with parasite resembling P. malariae 101 x infected with parasite resembling P. vivax 101 5s Pathological effects pro- duced by Strongyloides Tak) Wesaceaccneeeacccncn 283 Choanotaenia decacantha .......1.c10e00e 367 x microsoma, M.Sp., mor- phology ...... a ? octocantha 5 ? ungulifera Cholera possibly transmitted by cock- TOAChES s2:22 sfansaseastecmeeee rete toes 442 Gittotaenta -auicolaiests.cnctssenvecetse sees 147 5 bursaria Ss MOSAICa .. schtaen Civilian population of Malta, Un- dulant fever*in' > ©..iss.eeetesesceere II Clark, E. J., and Adler, Intra- uterine infection with 4. caninum IN GOQS «ssccsicesscsssncassscawsssassesseaes 353 Cleobonnea occulta ........cseeceeeeeeeeneees 321 Cockroaches, Réle of, in disease......... 441 Colouration, Influence of, on biting of MOSQUITOES He -ceeeeee merece coenee ee aenens 232 Corson, J. F. The occurrence of the larvae of O. volvulus in the skin of natives of the Gold Coast ............ 407 Corson, J. F., and Macfie, J. W.S. A new species of filarial larva found in the skin of natives in the Gold Coast 465 Corson, J. F., and Macfie, J. W. S Observations on Onchocerca volvulus 459 Cotugnia ? bifaria ... - 145 » fastigata 366 43 MAN BATE ainiondesissstietin wstoeene 147 53 oligorchis, sp.n., morphology 55 Craw-craw associated with infection with, OQ} voles esecesssdineseese soos 408 Grows; 'Cestodesiinemn) 22....0..2<...+- 146 Cryptocotyle lingua in a dog in England 153 Ctenocephalus canis from dogs and cats in) AMAZOMASHI Nee Sec cetlist weccsscossiess 298 Culex annulioris, female genitalia ...... 177 9). CBT SCROLUMIN Teteeicteesin ... ROBic canoe se eee es campanulata Be x centropi, 0.Sp., morphology 363 = ceylonica 3 CODM 1. ccsvewussuesccsedeetenetettee Pr columbae 3 CONOPOPhiled — .i.eeevsseceeneeee 144 5 COTWING sp coussSeeeteadutene oeeeee 146 i CTASSUIAS)... Sueeen onaneen ee eee 142 3 CTUCIALa Fisted Berea ooeenenee 358 > cryptacantha ...... a F dubius, diagnostic points sess 362 5 dublanensis, diagnostic points 362 5s fithrmanni, n.sp., morph- OLOGY! 15. .e be. onthe meen 137 ¥. GOUT Asa coves essav ees cne- thee eeeeee 142 Davainea himantopodis, diagnostic points insignis micracantha microscolecina minuta, diagnostic points ... MUMLGDELIS vou anne seca teats tek PAPATAGISED Se staat thabes ” ” Dipylidium caninum from dogs in Amazonas ...... ‘5 » from dogs and cats in India and Egypt...... 5 > as host of 4. cani- Teds aes voxer 9 35 WARUUL SE. ear eare 33 wos. «xs Dirofilaria immitis from dogs in Pana OAs sreatiaen dens sstaathi aca s asians Disease incidence in population groups Dog, Cryptocotyle lingua in a .....000. Dogs, Ancylostomes from, in Freetown Intra-uterine infection with 4. Cominune in), ee eee Parasites from, in Amazonas ... Pulmonary lesions in, naturally infected with nematodes Drepanidotaenia fasciata ” ” ” PWD SPU orn sans «sneenakidewaaes «ai 3 Bev acasccnmeslte 60, 146, PAGE 358 362 142 356 362 359 142 378 PAGE Drepanidotaenta gracilis .....2..000e0000s 145 mMegalorchis .......000+ 149 Ducks, Cpxtaitles fecha il deceit ta 8. 145 Duthiersia fimbriata ..cccccccsseseecevccees 150 Echinocotyle uralense ..1..cccsceecseeeeees 377 Echinorhynchotaenta nana, n.sp., morphology ... 193 tritesticulata ...... 196 Elephantiasis associated with infection with O, volvulus ...... 408 6 in cases with and without MicroMlariasseccres. +e 110 Emetine, Degenerative effect of, on Sa BASH ALO SUITS ccasin sarge dnans aos Seeaaie 209 Entamoeba blattarum from cockroaches 443 coli in Amazonas and else- Where My iratess desc aes 94, 97 »» cysts carried by cock- TOaChes< sdesades t-te histolytica in Amazonas and elsewhere 94, 97 cysts carried by cockroaches ... Relative portion ‘ordinary ’ and ‘small ’ strains nana in Amazonas and else- WHELE) menses esciadeadecgsss 94, 97 Eretmopodites chrysogaster, female geni- talia quinquevittatus, female genitalia Eukraiohelea foyt, n.sp., morphology... Eumelanomyia inconspicuosa, female Genitaliagumese tas. pt fo-once annette aztee Evans, A. M. Notes on Culicidae in Venézuela, with descriptions of new Species amu bact Ny infectioMIn: +25-6...-<-.5 460 n » New species of filarial larvakiromie™:.. .ccvecseees 465 Gordon, R. M. Ancylostomes recorded from sixty-seven post-mortems per- formed in Amazonas ..........-+.000+ 223 Gordon, R. M. Notes on the biono- mics of S. calopus in Brazil............ 425 Gordon, R. M. The occurrence of ancylostomes resembling Necator americanus amongst domestic pigs invA mazOnass. c.sadeeese facto tease tees Gordon, R. M. The susceptibility of 205 the individual to the bites of Stegomyta cAlopus ...........eceeeeecneees 229 Gordon, R. M., and Evans, A. M. Mosquitoes collected in the Mandos region of the Amazon ............... 315 Gordon, R. M., and Young, C. J. Parasites in dogs and cats in Amazoniasssscanseetaw secon somaces.Reeeae 207 Gregarina blattarum from cockroaches Gymnobhelea, alleged generic character of eyes Gyrocoelia australiensis, morphology... vill Haemoglobinaemia in case of black- WARES PALL Goo geonnas 333530330 seAC oo 453 Haemagogus €QuUinus .......00ccececeeceenes 327 Hairiness, Influence of, on biting of MOSQUITOES. -.0.5<. cceve ces eee eee 232 Hernia in cases with and without microfilaria,, ..s<.%0.22.9:cse eee eee Ill Heterodoxus longitarsus from dogs in AmaZONaS:\35a..-.0e-sedsce senate eeee eee 298 Hookworm disease possibly spread by cockroaches)... ..cssesasteeee eae ee eee 446 Human intestinal protozoa in— Amazonas... <: Neumanni », oblongiceps....... trigonophora Monobelea nigeriaé, 0.sp., morphology Moody, L. M. Alastrim; or Kaffir TO MPO KS eee eee see eeece essere seceereces 21 Mosquito, female, Genital armature of 157 0 repellents, Native ............ 438 Mosquitoes collected in the Manaos region of the Amazon = Individual susceptibility to the bites of Mucidus scatophagoides, female geni- ttalias ats Aswosdosnngnsdeaspsousenstohs feeds Nationality, Influence of, on biting of TMOSGUItOES aoe epee a heeas aa eeneaer Naval population of Malta, Undulant IRELZEIR a Bea -css85 9950005251070 TOIL IOS II Necator americanus amongst domestic pigs in Amazonas Eggs of, carried by cockroaches from 315 A 53 man, in Nematodes, dogs and. cats naturally infected with, Pulmonary lesions in Nematoparataenia paradoxa, 0.g., N.Sp., TOrpholo py cea ceeeeeet-ing vein ecunsieens Nematotaenia dispar — .r...0eeeeeeees 152, 192 Newstead, R. A new species of Phle- botomus from Trinidad - PAGE Newstead, R., and Evans, A. M. A new tsetse-fly from the South Cameroons 51 Nodular leprosy in the Sudan .......... 341 Notes on Australian cestodes 55, 61, 189, 305 » the bionomics of S. calopus iny Brazil iy) ,..s-xseneeeaes .-389, 425 » acase of blackwater fever ... 451 5 Culicidae in Venezuela, with descriptions of new species. Part, [3-004 Sota ceteeeeee 213 Ochlerotatus albocephalus, female geni- talia/...::).c.ceceaeetpeeeeee 169 38 apicoannulatus, female genitalia ...2c-:-..5ssseee 169 5 domesticus, female geni- talia.$;.::.<:-a.c0caeeeeeeee 169 = irritans, female genitalia 170 33 punctothoracis, female genitalia Oil as a larvicide : Onchocerca caecutiens, inability | to live in blood | 2oees. ss sseeeeseee 410 5 volvulus, experimental feed- ing to tsetse-flies 462 95 » infection, Diag- nosis Of ........-++ 459 25 » infection, incid- ence in Gold Coast. .vszita. ste a0 55 a5 larv ae, “ise tion in body 460 a oo. latwae;y gin athe blood sstscpeeeaxces 410 3 55 larvae in the skin of natives of the Gold Coast ...... 407 Pr sa larvae, Vitality of 410 Ophidotaenia natae ....... be See eGe Ophiotaenia ? calmetti Ps PUNICA! wsacesdssuea eae . 134 Oxyuris blattae from cockroaches ...... 443 Palpomyia pistiae, n.sp., morphology... 274 Parabezzia poikiloptera, n.sp., morph- ology 276 Ps 55 5) 9 lara gee pupa... 280 Parasite resembling P. falciparum in a chimpanzee: ....s.s.+..se First record of, from West Indies ............ sy habitstotl} tee. .ceeket. i282. é, habitatsiol. Ss2eii 2c a New species of, from iMninidads sezkeee Bees 33 preventive measures ...... = Seasonal prevalence of ... “3 minutus, inefficient vector of sand-fly fever ........ 3 papatasii, Importance of, as carrier of sand-fly fever 5g 2 in North of rance}2 ss. — : preponderance over other species in Macedonia... 3 perniciosus, as carrier of sand-fly fever ............ aS trinidadensis, 1.sp., mor- j2)(0) Cofy ARP Bere seca. Physopsis africana, Experimental infes- ARTO Glen seseers core ieceer otad-et aeas> Pigeons, Cestodes from Pigs, Necator americanus from, in BNINAZOWASs=scsacccsansusacteeeeareeset owes. ROACHES s lop 65055cee...cecdosvasteabentnes as Plasmodium falciparum, Parasite resem- bling, in a chimpanzee % inui, Parasite resembling, in a chimpanzee ......... ¥ malariae, Parasite resem- bling, in a chimpanzee.. s ovale, n.sp., morphology a vivax, Parasite resembling, in a chimpanzee ... = var. minuta Population groups, Incidence of a PISCASENIT cw soscvisnsies SESE CII a Sees Pott’s disease, evidence of occurrence, ZOOO B.C. vecccscesscccscrccccevcccescucces Prevalence of Ceratopogonine midges on the windows of the Accra Labora- tory during a completed year PAGE | xi Prionognathus, pre- Ocanpied Iereieats. tacs.tod.weeh osetia: Protozoa, Human intestinal, in— Amazonas generic name ipayyith aeatete toscctee 2 oocoscentesnanncase England........ Vic leeds eeaceee neki onet Ge ay estas Ca. <6 Queensland IESHAL Seets pos mlseteieteces ee bre etasee ote. Psorophora ctliata.......ccccseveeseeeeerecees 35 dubia eerste OSE 218, es posticatus posticatus.........+- $5 a AY Ieetaeces > SAERU scaler se edenics ue onesstee Soe tovart, n.sp., morphology... Pulmonary lesions in dogs and cats naturally infected with nematodes Repellents, Native mosquito Reptiles, Cestodes from Rhabdometra tomica Rhipicephalus sanguineus from dogs in ANTAL ODA! RSME Saeee Teese ov eweceee Sabethes amazonicus, sp.n., morphology 3 LR eee Sabethinus undosus..........010+c0ecseereseee 3 > $5 aE Wate a eaten 3 pupa Sande ‘Ay, First record of, from West LG SS | oe Renee eR Re eae ae 3 fever, Relative importance of carriers of Sand-flies, Bionomics of ..............0++ Schistosomes infesting fresh-water (Siti cra day 22 eno ee Schistosoma haematobium, Degenera- tive effect of emetine Ove: Seas 35 + Eggs of, carried by cock- roaches... Schistosomum Bovis ..ccciccecevececeesececes Schizodactylus, generic name _pre- OCCU PLSHrnee nei Bece « .eeIentetacs ovens Schizotaenia cacatuae, sp.n., morph- OB YrasedadesesescasacecsesAstehvoasssenase 7 Sex factor in cases with and without microfilaria or signs of filarial disease Sierra Leone, Filarial disease in Natives of PAGE 207 209 Skin, Larvae of O. volvulus in the...... 5, New species of filarial larva found an ees Pee Sleeping sickness, Case of ............... Fe 3 » treated with ‘ Bayer 205” i 3 T. rhodesiense in a case of, from the Sudan Messen Southwell, T. Cestodes in the collec- tion of the Indian Museum ......... Southwell, T. Cestodes from Indian birds, with a note on Ligula intes- EIMQVIS, ccwsicazs saps osadcsodeeneee ess =6en Southwell, T., and Maplestone, P. A. Notes on Australian cestodes ...61, Spirochaeta sp. from cockroaches ...... Spirochaetosis, Bronchial, simulating pulmonary tuberculosis...............+. Staphylococcus aureus carried by cock- TOAChES:aeensc 2s eee fi citreus carried by cock- TOACHESissecka tacos eee Stegomyia apicoargentea, female penitalia@eeeetess x calopus, bionomics of, in Brazilesccse 389, cr ,, biting of cadavers... s » breeding during WOVAVEMEL cseeten ss 5 », conditions of laying and hatching of CRE S heey ae yasteee! A », development of larvae and pupae es 5, Individual suscepti- bility to the bites Ole th RAS EF », larvae absent from natural waters ... - ;, Natural enemies of “ 5, order of hatching of males and females aS 57) CONMLatIOnN Ss hetesnes 3 dendrophila, female genitalia we fasciata, female genitalia... 53 luteocephala, female geni- Pali ae ere cs cseraceee eee = metallica, female genitalia i simpsont, female genitalia ... 35 unilineata, female genitalia 3 vittata, female genitalia...... 425 435 391 399 229 425 436 429 431 168 166 168 169 169 169 169 xii PAGE Stephens, J. W. W. The incidence of a disease in population groups, the number of people in which is known (OT UK O Wilts eeescacce sce seeceeeeee eee naem 199 Stephens, J. W. W. A new malaria pavasite Of anal ess... seddeensee eee 383 Stephens, J. W. W. Undulant fever in the naval, military, and civilian populations of Malta.............sece0es II Stephens, J. W. W., and Yorke, W. Case of sleeping sickness (J. gam- biense) treated by ‘ Bayer 205” ...... 421 Stilesia globipunctata ........s.ccs.ceeeeee 132 Stilobezzia limnophila, n.sp., mor- phology. seso-2ecceesceecce eeee ns eeeeeeee 267 Strongyloides larvae in faeces of chim- PaDZeesc, » situberculosis ini recesses receee 235 Susceptibility of the individual to the bites! of S.. calopus\scesstavncasneeneenanere 229 Sweating, Influence of, on biting of INLOSG WL OSs. oeneaee sere ees eee sseeereee ene 232 Tenia AMMONILIPOTMIS .....00.ceeeeeeeeees 128 jo OGAT ALE x vcs escwacewseseeeeneeeee eee 145 - COLUG. o ccsiaava sidatoak se estheneaen eee 147 330 Carolts....ciee eee 148 55 CL ASSUCOLLES. csc sca 5s cn deo eee eee 127 39, > we Oligartlriztsc..ececsek- -eeereae eee 128 5, 0 delafondt.-... nesacenste ane eee 148 9). Mn PISU/OrMmisA s.ceseseesteeeeeeeees 127, 129 gn polycalcartic hn. .eeteeeenceeeereeeee 128 » saginata, Eggs of, carried by cockroaches) ts.3. ahsoeeeeaeeees 447 £5 8 SEPPGLGON ks. aoe sees 127, 128 5; se Sphenoceph alas wensenteneeseeeeees 148 ‘ TO AL DA tns nees oe Peete nee 147 Taeniorhynchus aurites, female geni- alia’ Wissasaceecee conean ese seen apee eee 174 Testis, Enlarged, in cases with and without microfilaria.............sseee0e 110 Tetrabothrius rostris .....cccceceeeseeeees 356 Thysanognathus, new generic name ... 244 +. albopictus, n.sp., mor- phology ...........06 244 5 melanostictus, N.Sp., morphology.......... 248 Toxascaris from cats and dogs in Bxeetopyniccs.coss. scenes 292 SS canis from dogs in Ama- ZONAS\ . sccscresessensenarenens 298 Toxorhynchites brevipalpis, female, genital armature Of) ...........000000 164 Trichodectes latus — dogs in Ama- © cL) ocagen Sa Saacsbecodososceessssorumeene 298 Trichomonas hominis in Amazonas and elsewhere 94, 97 Trichuris ova from a dog in Amazonas 299 trichiura, Eggs of, carried by cockroaches 3; Trinidad, New species of Phlebotomus REN Ss sstupn cwwcswearsaas Mute Te. Sit ose Trypanosoma gambiense, Case infected with, treated by ‘ ——— 205° eeecncctberseedeessus rhodesiense in a case of sleeping sickness from the Sudan Trypanosomiasis, Case of ..........:.00004 Tsetse-fly, New, from Cameroons ... Tsetse-flies, experimental feeding with O. volvulus Tuberculosis, Pulmonary, simulated by bronchial spirochaeto- ” 447 inthe; Sudan | sccecascseeces 3 Young, C. J. Notes on the bio- nomics of S. calopus in Brazil... 389 Young, C. J., and Gordon, R. M. Parasites in dogs and cats in Ama- DIG AS Mea eat oesione sc saasen swelas tar 207 Zammit reaction for milk.............006 3 Zammit, T. Undulant fever in the BOMB Wiad) .vsomeacigneecwavasconeerc I INDEX OF GENERA, SPECIES AND VARIETIES NEW TO SCIENCE A édes oswaldi var. braziliensis ......... Agamofilaria streptocerca......1...s...000+ Ankistrodactylus par .......e0eseoseeceennee Anopheles (Arribalzagia) venezuelae ... A trichopogon acan EOCOI UAT Rceeatenee se te A trichopogon ACOSMELUM ....0..c0e0esee00ee A trichopogon hespertum ...cccecerseeeeeees A trichopogon kelainosoma ....1.....00.000- Choanotaenia Micr0soma ...cececececeveeee 3 Cotugnia oligorchis .........0cc0seeeeeeenees Culex originator . Dasyhelea flavipic Dasyhelea omoxan Davainea centropi URS ABA RDA CECE RE HOHCAtIO tha basse eaaaviedeR Davainea fubrmannt ......0000.c00eseeeeee Davained tragopant ........00ecerseceneeene 7 Echinorhynchotaenta nana .....1..e00ee00s Eukraiohelea foyi xiv PAGE Glossina hamingtont ....00...s0cceeeseceernes 51 Hymenolepis annandalet........10c0000000 374 Megarhinus boret .....c0.cssscssscvessesnsve 330 Monobelea nigeriae ............0+.er0seore 268 Nematoparataenia .....sceccecvecevecreeeees 193 Nematoparataenta paradox ......+00006+ 189 Palponiy iG Pisti ese an cneeoaneeese sa seeeneee 274 Parabezzia potkiloptera.....s.s.secceeeeees 276 Phlebotomus trinidadensts.....1..s0ss0e00e+ 47 Plasmodium ovale ........ecescoweseenseee 384 Psorophora (fanthinosoma) tovari ....... 218 Sabethes AMAZONACUS.....0...0eeeeseeeeseeens 316 Schizotaenta CACAtUAE ....4..0..0..00eeeeee 305 Stilobexzta limnophila.......c0.cceeeereeees 267 Thysanognathus albopictus ........0000006 244 Thysanognathus melanostictus ......+++0+ 248 Uranotaenia calosomata var. albitarsis 335 WY Comytd NERTENSIS.....ce0eccnnssnnercnsses 319 Proressor A. RAILLIET UNDULANT FEVER IN THE GOAT , IN MALTA BY ae (ZAMMEE ( COM,G. oN. D. GOVERNMENT ANALYST, PUBLIC HEALTH DEPARTMENT, MALTA (Received for publication 17 October, 1921) PLATES I AND II Undulant fever, which, notwithstanding the decisions of Inter- national Boards, is still by many called Mediterranean fever or Malta fever, is, as heretofore, a disease which causes much suffering and anxiety. Since the year 1905, when it was demonstrated that the infection was caused by the drinking of milk, more especially that of goats, the Public Health Department of Malta has been endeavouring not only to minimise the occurrence of the disease, but to find a way to stamp it out. For this purpose it has been instructing the public as to the prophylactic measures it should follow, and at the same time keeping watch over the milch animals on the Islands. In the ten years (1894-1903) preceding the appointment in 1904 of the Mediterranean Fever Commission, the average number of cases in the Maltese Islands was Malta 3:2, Gozo 1'9 per 1,000 of estimated mean population per annum. During the years 1901, 1902 and 1903, among the ships of the British Mediterranean Squadron constantly at Malta, with an average crew of 8,230, there was an average of 28°55 cases per 1,000; whereas from 1897 to 1903, amongst the British Garrison on the Islands, there was an average of 25°6 cases per 1,000 per annum. The apparently greater rate of infection among the Services, as compared with that of the civil population, may be explained by the fact that in the Services every case of illness comes under the notice of the Medical authorities, whereas in the civil population slight cases pass unnoticed, and other, possibly numerous cases, are either incorrectly reported or are not reported at all. This happened more especially before 1904, when, owing to insufficient knowledge, the fever was incorrectly diagnosed, and sufficient importance was not given to it. 2? < When it was found out that the fever resulted from the drinking of goat’s milk, and that the micro-organism causing the infection could be destroyed at a temperature below 100°C., the Maltese Sanitary Authorities at once made it known that the heating to boiling point of fresh milk would free it from infection. The Naval and Military Authorities were also prompt to take action. Thus in June, 1906, goat’s milk was banished from the dietary of the Garrison (A./.D. Report for 1906, Vol. XLVIII, p. 78), while the C.-in-C.’s General Order to the Fleet, dated 4th August, 1906, prohibited the use of unboiled milk (Navy (Health) Statistical Report for 1906, p. 119). By these means a disease, the etiology of which had not only baffled the skill of medical men for about a century, but had also affected the efficiency of the Mediterranean Fleet and the Malta Garrison, was arrested ; probably a unique case in medical history. The use of fresh milk had no sooner been tabooed in the Services than the number of cases of undulant fever dropped as if by magic, and both the Navy and the Army on this station have since been almost entirely free from it. English people, whose duty compelled them to reside in Malta for some time, obtained a like relief by adopting the simple precaution of boiling the fresh milk or abstaining from it. Before 1904, few of the employees of the Electric Telegraph Company, the dockyard, etc., or their families, escaped infection; but it is now an exception for a foreign resident, who takes the necessary precautions, to fall a victim to undulant fever. The bulk of the Maltese population, who thought they knew better, have not heeded the caution repeatedly given out by the Sanitary Office as to the danger of using unboiled milk, with the result that there has been hardly any decrease in the number of cases of undulant fever. THE GOATS “The Maltese goat is the hardiest, the tamest, the best milking goat in existence. It bears a resemblance to the Theban or Egyptian goat, from which it probably originated. Like the Theban goat, it is generally beardless and frequently hornless, has spreading and slightly pendulous ears, though shorter and narrower, has a convex profile though not so marked as in the Theban goat, 3 has very often a pair of lappets on the throat, and like it is often of a reddish colour, but it has larger hair and the udders are very large, in relation to its remarkable milking qualities. Maltese goats milking at the rate of 5} litres (about 9} pints) in twenty-four hours are not uncommon. White haired goats were formerly preferred by goatmen, but it was found that they are less hardy than the reddish or black-haired ones.’ (Dr. J. Borg, in ‘Malta and Gibraltar,’ compiled by Allister Macmillan, p. 237, London, 1915.) From time immemorial, goats in Malta have been milked at people’s doors, and it is impossible by legislation to compel house- holders to boil their milk, but it has been made unlawful for hotels, restaurants, coffee-houses, etc., to serve other than boiled milk. Such a measure should have helped to educate the people in the matter, but the general public has yet to be convinced that an apparently normal beverage drawn straight from the familiar goat can be productive of a deadly fever. One must also bear in mind that the conclusions of the Commission have not remained unchallenged ; apologists have come forward offering negative evidence in defence of the offending goat. Others, who could see only the. financial side of the question, have pleaded the cause of the poor milkman. The Sanitary Authorities, seeing how their efforts were being thwarted, devised other means to protect the sceptical public. Periodical inspection of goats by trained sanitary officers was instituted, and samples of milk, or blood, were taken. The Widal and the Zammit* tests are applied to the blood or milk respectively on the day of their collection. If the goat is found to react it is sent to the Lazaretto under escort, and is there examined by a veterinary surgeon, who assesses its value as a dry goat. The * Widal reaction. Dilutions of the serum of 1 in 80 and 1 in 100 are made with salt solution. The emulsion of AM. melitensis consists of one to which formalin has been added. Onc drop of the emulsion and 1 drop of the diluted serum are mixed. The mixture is placed on one of the 4-inch spaces ruled with a diamond on a glass slide about a foot long. The slide is rocked to and fro for about a minute and placed in a wet chamber at room temperature. Results are read after about two hours, but the reaction is usually obtained, in a positive case, after one minute’s rocking of the slide. Naked-eye appearance is sufficient, though a hand lens is sometimes used. In this way about too specimens of blood can be examined in a day. Zammit reaction for milk, Loopfuls of diluted milk and emulsion of culture are mixed on a slide so as to give resulting dilutions of 1 in 20 or 1 in go. The mixture is drawn into a capillary tube, which is sealed at both ends and stood on end in sand. The result is read next day, though the reaction is sufficiently clear in a couple of hours, the naked-eye appearance of the precipitate being quite characteristic. In doubtful cases the previous test is applied. 4 owner has the right of appeal on the question of value. The goat is slaughtered at the Lazaretto. , The average annual amount paid as compensation to owners for the destruction of infected goats is about £500. In the estimates for 1921 the sum of £700 1s allocated. The average cost of an infected goat in pre-war time was 20s. It was primarily intended to inspect the twenty thousand odd goats spread over the two islands twice yearly; but the limited special staff available could not cope with the work, and the inspection of goats, sheep and cows is consequently restricted. As the inspections are made periodically only, animals that become infected in the interval remain undetected for some time, and consequently a number of infected goats always exist; hence it is not surprising that undulant fever is still prevalent among the civil population. The systematic, but limited purification of the herds effectually reduced the disease among the animals, and although the frequent examination of every milch animal, with the consequent slaughter of those found infected, is costly, it should be made continuous if the fever is to be eliminated. A much larger staff should be organised; that available has been unable to inspect yearly more than about six thousand, out of about twenty thousand goats. According to the reports of the Public Health Department, the number of goats examined and the rate of infection found were as follows :— Taste I Showing the rate of infection with AZ. melitensis of goats in Malta ! Year | No. of goats .|-. No. infected Percentage | examined ae | | 1907-08 a. oo non 300 rod| 1,203 170 | I4'I 1go8-09 ... ed ae 2 <3] 1,099 2 2'9 1g0g-IO ks a sr ont one 9,924 461 | 46 IQIO-I1. « sar aA 250 oe 13,372 402 30 IQII-12; «.- ‘= bon = ae 13,756 386 28 LQUZ-13) ine aes * a aI 11,453 414 3°6 Igl3-I14 we “os ocd a as: 6,896 381 5°9 IQI4-15 + see oe cos Ae ee 45965 385 77 195-16... ote cen ooo “ee 6,630 | 598 go 1916-17 ~ «.. o “on ote cect 75768 536 6-9 1917-18 sn evs se oes 59921 287 48 1918-19. ..- ee ate aie See 4,613 187 4°70 I9IQ-20 «se sas tes ve core 5,690 | 341 59 | 5 The infection rate is consequently about 5 per cent., a dangerous percentage, for a single goat may infect hundreds of persons during its milking activity. Sheep are less liable to infection than goats, probably owing to the smaller size of their udders with a consequent smaller chance of abrasion, but there are no reliable statistics as to the number of sheep infected. . In the light of modern treatment of infective diseases, prophylactic inoculations with a smelitensis vaccine have been suggested on various occasions. In 1906, Dr. Eyre, one of the members of the Mediterranean Fever Commission, used a vaccine in fifty-one cases; of these, twenty-two received one injection and twenty-nine received two injections of 200-400 million cocci. Two of the cases vaccinated contracted the disease. Professor M. H. Vincent (1918), of Paris, carried out vaccination experiments on goats and published his results in a paper, in which he claimed to have solved the problem of the melé/ensis infection. - The Maltese Government, wishing to utilize Professor Vincent’s vaccine, asked the writer to report on the matter. It was agreed, therefore, to repeat Vincent’s experiments as described by him, on a number of local goats. The writer, who was no longer in a position to conduct the experiments himself, had the honour to be entrusted with their supervision. The Technical staff of the Public Health Department carried out the experiments in a most conscientious manner. They all had long experience, both with the micro-organism and with infected goats, and followed Vincent’s directions in all particulars. A full report of the work will eventually be published, so that I will only mention the broad conclusion arrived at, that is, that the bright hopes built on the French savant’s paper have been dashed to the ground. The immunisation of the vaccinated animals did not occur, and a minimal dose of virulent culture of M/. melitensis infected both the experimental animals and the controls. The question remains therefore, 77 s¢a/w guo, and either another vaccine will have to be devised or vigorous direct action be taken to free the island from the fever. At this point, one cannot allow to go unchallenged an assertion that Professor Vincent made in the above-mentioned paper to the 6 effect ‘that an infected goat recovers spontaneously after a period of time more or less long.’ This is a bold assertion, which is intimately connected with the whole prophylactic question of the fever. I do not believe that Professor Vincent is justified in making such an assertion, which is contrary to our experience. The writer who has a very long experience of goats, both normal and infected with 1/. melitensis, has never known an infected goat to recover. The animal may feed well and look bright, its blood may completely lose its agglutinating power, but a careful post- mortem examination shows, as a rule, the micrococcus lurking in one or another of the glands. After kidding, a goat, that has for about two years appeared healthy and free from an infection it had previously contracted, yields a milk teeming with melitensis. It is satisfactory to read in the Chief Government Medical Officer’s Report for 1918-19, that ‘the notified number of cases (three hundred and sixty-three) points again this year to diminished incidence of the disease, with deaths—sixteen—representing a case mortality of only 4°4 per cent. ‘This is the smallest on record, and is equal only to a death rate of 7°1 per 100,000.’ In the report of 1919-20, however, the disease shows a slight recrudescence (six hundred and nineteen cases with a case mortality of 51 per cent.), which reduces the hope for a progressive amelioration. The civil population, too, can protect itself, by using no milk that is not previously boiled. Were this simple procedure to be strictly followed, the fever would disappear from the civil population as it did from the Navy and Army. As, however, most of the people remain obdurate, or careless, it is our duty to eradicate the disease by vigorously acting against the main cause: by destroying every animal found to be infected. Goats are not susceptible to a cure, and even were a cure possible, their treatment would, in the happiest event, be long and costly; more costly than the animal itself. An adequate inspection staff should be provided, and no expense spared. Were this done, I am confident that the fever would disappear from the island in a short time. REFERENCE Vincent, M. H. (1918). Sur la prophylaxie de la Fiévre de Malte par l’immunisation active des animaux vecteurs du germe. Comptes Rendus de l' Academie de Science, Feb. ' ce STs: WO YVONTAVYVAINZSA , . ul Hi mira aT std ie) duphyeertyy 263) ; ape fe eG aitiahy biewien f 4 EXPLANATION OF PLATE I Fig. 1. Maltese Goat, presented to the Museum of the Liverpool School of Tropical Medicine by Prof. T. Zammit. Photo by Miss M. Brown. Fig. 2. Group of goats, Malta. Maltese Goat, presented to the Museum of the Liverpool School of Tropical Medicine by Prof. T. Zammit. Photo by Miss M. Brown. Fig. Ge Annals Trop. Med. & Parasitol. Vol.:XV 1 PLATE 1 ; C. Tinling & Co., Ltd., Imp. * , ‘ 2 a ae) i oe me — be RV rite _ pels 7 Io EXPLANATION Fig. 1. A milch goat, Malta. Fig. 2. Group of goats, Malta. Fig. 3. A milch goat, Malta. OF PEATE it Annals Trop. Med. G& Parasitol., Vol. XVI PLATE J] C. Tinling S Co., Lid., Imp. II UNDULANT FEVER IN THE NAVAL, MILITARY AND CIVILIAN POPULATIONS OF MALTA BY J. W. W. STEPHENS (Received for publication to February, 1922) In the previous paper Zammit has considered the prevalence of undulant fever* in the goat in Malta. As complementary to that paper I thought it would be of interest to present afresh the data as to the prevalence of the fever in the Mediterranean Squadron, Army (Malta Garrison), and civilian population of Malta respectively previous to 1906, and from that time onwards so far as data are available. 1906 is the critical year in regard to the relationship of undulant fever and goats’ milk, for it was mainly in the latter half of that year that orders affecting the use of goats’ milk came into force. I have prefaced each section of the paper by some remarks, with the object of elucidating the meaning of the figures presented; for it is difficult, in the case of many, if not all, vital statistics, to ascertain whether they really represent what they are supposed to do. In the present connection it is essential to know whether a case ‘ returned’ as undulant fever is that fever or not. Undulant fever is among the select class of infections that can be diagnosed with certainty. It is probable not only that certainty has not been attained in many cases, but that the approach thereto is a variable one. NAVY MEDITERRANEAN SQUADRON 1. NOMENCLATURE : The following terminology is used in the ‘ Statistical Reports of the Health of the Navy ’ :— Other Continued Fevers is used from 1900-1906 and signifies fevers other than enteric and Mediterranean, influenza appearing under its own heading. Pyrexia replaces ‘ Other Continued Fevers ’ in the tables from 1907-1914. “In the ‘Nomenclature of Diseases,’ Royal College of Physicians, London, 1897, 3rd Edition, appears the entry, Mediterranean Fever, synonym Malta Fever. In 1918, 5th Edition, the entry, Mediterranean Fever, synonym Undulant Fever is used. 12 Simple Continued Fever is also used synonymously with Pyrexia in the text in 1909. Sand-fly Fever. In the report for rgto0, p. 50, it is stated that of the 98 cases of ‘ Pyrexia’ in that year 58 were ‘ Sand-fly Fever.’ Cerebro-spinal Fever has a separate heading in 1912. Mediterranean Fever is used in the tables from 1900-1906 and from IQIO-1914. Malta Fever is used in the tables and text from 1907-1909. 2. DIAGNOSIS: 1g00. ‘ Agglutination adopted as a routine practice in the Army and shortly after in the Navy.’ (Report of the Commission on Mediterranean Fever, Pt. II, 1905, p. 12.) 3. Goats’ MILK: (a) Naval Hospital. 1g06—April 9. ‘ More stringent measures taken at the Royal Naval Hospital for the sterilisation of goats’ milk.’ 1906—July 23. ‘ Preserved milk substituted for goats’ milk.’ (Statistical Report of the Health of the Navy, 1906, p. 120.) (@) lane ilaee. 1906—May 23. ‘ The Commander-in-Chief promulgated a general memorandum to the effect that as a guarantee of sterilisation the ortol and peroxide of hydrogen test should be used in all ships.’ 1906—Aug. 4. ‘ The Commander-in-Chief repeated the order that all milk obtained in Malta was to be boiled. It was then to be tested by the ortol test.’ (Commission Report, Pt. VII, 1907, pp. 72 and 73.) 4. EstIMATED STRENGTH : The figures for the average strength refer to the Mediterranean Squadron and not simply to Malta, so that the rate per 1,000 is not comparable with that of the garrison or civilian population of Malta. . These figures are corrected for time, i.e., if 1,000 men have been in the Mediterranean Squadron for six months and 500 for one year, the average strength per‘annum is 1,000: or again, if 365 men have been in the Squadron for one day and one man for 365 days, the average strength per annum is 2. ; ; 13 ‘Taare I Showing prevalence of Undulant and certain other fevers in the Mediterranean Squadron, 1900-1914 Average Cases Rate per 1000 per annum strength Sa ee Year corrected Other Other for time | Mediterranean) continued | Mediterranean | continued fever feyers fever fevers TQOO a oe 14250 | 317 351 22°2, 24°6 IgOl bec 14070 | 252 323 17°9 22°9 BGO2: «<5 ae 18470 | 354 433 1g't 23°4 1903 |. S8gq0 339 287 184 15°5 1904 19590 | 333 401 1770 20°4 1905 | 14360 270 174 18°8 121 1906 | 12130 145 99 11g Sr | | | Pyrexia Pyrexia 1907... «es | 10530) | 14 110 V3 10° 1908... | 9780 6 119 o6 / 12'T 1gog ... An | gg20 | 1t 69 Vi 69 I9lO ... ap 9850 3 98 o'3 9°9 1QI .. “| 9770 | 5 144 o'5 17 T9QL2 ise Seal 7870 | 3 49 o°3 Oz 1913 + ao 7580 | 2 38 o'2 570 BOTH. 6. we 10220 | 6 34 o'5 33 ARMY ! ! MALTA GARRISON _ I. NOMENCLATURE : ; The following terminology is used in the Army Medical Department Reports :— Other Continued Fevers is used in the statistical tables for the years 1897- 1907. Simple Continued Fever is used as one sub-division of ‘ Other Continued Fevers’ in the text from 1897-1907, the other sub-division being Mediterranean fever from 1897-1903, and Malta fever from 1904- 1907. Ve Pyrexia of Uncertain Origin (P.U.O.) replaces ‘ Simple Continued Fever ’ in the statistical tables and in the text for the years 1908-1914. Sand-fiy Fever appears in the tables and text for 1910-1914. Mediterranean Fever is used in the text from 1897-1903 and also from IQI0-IQI4. Malta Fever is used in the tables and text from 1904-1909. 2. DIAGNOSIS: 1g00. ‘ Agglutination adopted as a routine practice in the Army and shortly after in the Navy.’ (Commission Report, Pt. II, 1905, p. 12.) Major-General Sir William Leishman, K.C.B., F.R.S., has informed me that the diagnosis of undulant fever is always based on the agglutination reaction. 3. Goats’ MILK: 1905. It is stated that in 1905 ‘ orders were issued by commanding officers that all goats’ milk for the use of the men in barracks was to be boiled.’ (Commission Report, Pt. VII, p. 168.) 1g05—September. The attention of officers commanding was called to the fact that ‘in some corps goats’ milk had not been boiled before use ’ (p. 169). 1g00—May 16. The ortol test for detecting unboiled milk was in use and ‘during the next three weeks neglect of boiling goats: milk was detected on six separate occasions ’ (p. 170). 1g00—May 12. ‘Orders were issued for the discontinuance of the use of goats’ milk in the military hospitals as a tentative measure, and for its replacement by condensed milk.’ ‘This change came into operation in the various hospitals between May 18 and 22, and at the same time the use of goats’ milk by the various detachments of the Royal Army Medical Corps also ceased ’ (p. 172). 1g06—June. ‘ By the end of the first week in June all the units of the garrison were using condensed milk, with the single exception of the ist Battalion Rifle Brigade, which continued to use goats’ milk up to October ’ (p. 173). 15 Tasie IL Showing prevalence of Undulant and certain other fevers in the Malta Garrison, 1897-1914 Cases / j;-————__—_ ——————_|_ Rate per 1000 Year Average Simple | per annum strength Mediterranean continued | Mediterranean fever fever fever 1897 ..- i343 420 8023 279 1275 34°7 1898... oa cr 739° 199 1510 27°! 1899... see se 7425 275 1107 37°09 1900 ... = as 8140 158 1158 1g} Igor ... a ve $136 253 1205 } 3rI 1902 ... oe a $758 155 | 1029 17°7 1903... ‘8 as 8903 404 786 45°4 Other continued fevers Tgog ... “8 i gloz 320 1350 3571 1905... os oe 8294 643 1199 | 77°5 1906... ane =H 6661 161 508 24°1 1907... a2 rc 5700 Ir 5 323 19 Pyrexia of uncertain origin | 1908... ee aan 6030 5 303 } o$2 1909 | 6392 I | 285 O's Sand-fly | fever IgQlO ... oF aan 6769 ° . 26 124 | oo I9gtl ... sex oe 6686 ° 1y 125 | o'o 1gI2 ... ten one 6593 | 3 5 10g | O"45 1913 « eee see 6336 3 25 72 O'47 1914 (7 months only) 3487 I ° 5t 0°28 16 CIVILIAN POPULATION OF MALTA I. NOMENCLATURE: “ All fevers lasting more than a week are notifiable by law.’ ‘ Mediterranean fever is generally notified under the name of remittent fever.’ (Commission Reports, Pt. 11, 1905, p. 15.) It appears from the above Report that in the civil official notification returns, cases notified under the name ‘ continuous fever’ are included in the Annual Public Health Reports under heading ‘ Mediterranean fever.’ We find the following terminology employed in the Public Health Reports : Remittent Fever is used (for Mediterranean fever) in the Reports for 1897 and 1902-03. Continued Fever, in addition to remittent fever, or Mediterranean fever, is used in the Reports for 1897, 1906-07, and (apparently synonymously with febricula) 1907-08, and then disappears. Febricula, in addition to remittent, or Mediterranean, fever, is used in the Reports from 1898 to 1g06-07 and then disappears. The number of cases for the five years, 1902-03 to 1906-07, was 66, 35, 20, 22, and 42 respectively. Undulant Fever (for Mediterranean fever) first appears in the Report for 1912-13. The terms ‘Simple continued fever,’ ‘ Pyrexia of uncertain origin,’ and ‘ Sand-fly fever ’ do not appear in any of the reports. 2. DIAGNOSIS: That the agglutination test is in use for purposes of diagnosis appears from the Public Health Reports, for in the Report for 1912-13, p. 30, it is stated that ‘ 636 samples of blood were submitted by private medical practitioners for the agglutination test of cases of fever.’ 3. Goats’ MILK: 1907-08. The use of boiled goats’ milk adopted in the Central General Hospital. (Annual Report, Public Health Depariment, 1908-09, p. 5.) Igog—June. Regulations were issued apparently at this time requiring that all milk sold in shops, restaurants, etc., be boiled, but it appears from the Public Health Reports (1911-12, p. 43) that ‘it is very seldom that the law is complied with.’ a7: Taste III Table showing prevalence of Undulant fever and Febricula in the civilian population of Malta, 1902-3 BIOS 4: 1904-5 1905-6 1906-7 1907-8 1908-9 1909-10 IQIo-11 IQII-12 1912-13 1913-14 1914-15 1915-16 1916-17 1917-18 1918-19 1919-20 1902-03 to 1919-20. Year | Population 193,315 197,079 202,134 205,059 206,689 209,974 212,888 215,879 213,395 2155332 216,617 216,879 218,542 220,968 223,741 224,326 BA 224,655 224,859 Cases Undulant fever | 589 573 663 822 714 sor 463 463 297 275 37° 338 321 473 495 429 | 363 619 Febricula ** Continued fever.’ Rate per 1000 per annum Undulant fever 18 Per Mille. 20 10 1900 1901 1902 1903 1904 1905 1906 1907 1908 1909 1910 1911 1912 1913 1914 Cuart I. Showing incidence of Undulant Fever in the Mediterranean Squadron. N.B.—One division of this scale represents 5 per 1000. rg NILA PATNI WATT, 1897 1898 1899 1900 1901 1902 1903 1904 1905 1906 1907 1908 1909 1910 1911 1912 1913 1914 Per Mille. 40 Cuart II. Showing incidence of Undulant Fever in the Malta Garrison. N.B.—One division of this scale represents 20 per 1,000. 20 1902 1903 1904 1905 1906 1907 1908 1909 1910 1911 1912 1913 1914 1915 1916 1917 1918 1919 038 -04 -05 -06 -07 -08 -09 -10 -1l -12 -13 -14 -15 -16 -17 -18 -19 -20 Cuarr III. Showing incidence of Undulant Fever in the Civil Population of Malta. N.B.—One division of this scale represents 2°5 per 1,000. REFERENCES Army Medical Department: Report, 1897-1908 continued as Report on the Health of the Army, 1909-1914. Navy (Health) Statistical Report of the Health of the Navy, 1900-1914. Malta. Public Health Department: Report, 1902-03—1903-04 continued as Annual Report on the Working of the Public Health Department, 1904-05—1915-16 continued as Report on the Health of the Maltese Islands, 1916-17—1919-20. Reports of the Commission appointed by the Admiralty, the War Office and the Civil Govern- ment of Malta, for the Investigation of Mediterranean Fever, under the supervision of an advisory committee of the Royal Society. Parts I-VII. London, 1905-1907. 21 ALASTRIM; OR, KAFFIR MILK POX BY L. M. MOODY, M.D., B.S., M.R.C.P. (Lond.) GOVERNMENT BACTERIOLOGIST, KINGSTON, JAMAICA (Received for publication 19 December, 1921) Pirates III-VII For some months before my arrival in the Colony an epidemic of an eruptive fever described as Alastrim, or Kaffr Milk Pox, broke out in Kingston, and from Kingston spread to the other parts of the Island. From about May, 1920, to the end of March, 1921, two thousand nine hundred and twelve cases have passed through the Isolation Hospital at Bumper Hall, Kingston, and about six thousand have occurred throughout the Island. In the following paper I propose to consider :— I. The clinical aspect of the disease (p. 21); II. Its occurrence in the foetus (p. 29); III. Its relation to vaccination (p. 32); IV. Its morbid anatomy (p. 34). I, CLINICAL ASPECT Incubation period. Owing to the difficulty of getting cases in which exposure occurred only once, and that for a short time, it has been impossible to determine the exact period of incubation; but in those in which I was able to get some definite history of exposure the incubation period varied from ten to about fourteen days. The evidence on which this conclusion is based is as follows :— Case 1. A.H. Not vaccinated. He was in Kingston for three days during July when the epidemic was limited to Kingston only, and returned to his country district, where no cases had hitherto occurred. The symptoms developed to days after his return. Cast 2. Photographer E. Not vaccinated. He attended with me at the Isolation Hospital and took his first set of photographs on July zoth, 1920. During this visit he placed his focussing cloth, before using it, on a chair which had been previously occupied by a patient. Ten days later he again took a photograph, and 3 days after the 2nd photograph he developed symptoms. He was positive 22 that these were the only-two occasions on which he was exposed. Fortunately, his case proved to be mild in character. Case 3. Nurse at the Isolation Hospital. Not vaccinated. She complained of headache and pain in the back 14 days after she took up duty. So far as she knows she had not previously been exposed. The disease ran its usual course. This case had been exposed to infection several times before symptoms developed and is only useful as determining the upper limit of the incubation period. Cast 4. A boy at school in Kingston. He developed symptoms at Annotto Bay 11 days after leaving his school in Kingston. He stated that a number of cases of Alastrim had occurred at his school, but he did not know when he had himself been exposed to infection. He was the first case which occurred in Annotto Bay, and like Case 1, was directly traceable to Kingston. The remaining cases are not so definite. Casrs 5-7. During their stay at the Isolation Hospital a number of pregnant women suffering from Alastrim gave birth to children. Three of these cases I saw. The infants at birth were free of all signs of the disease. They were breast-fed by their mothers, and the rashes appeared on the roth, 11th, and 12th days after birth respectively. Not much weight can be attached to this evidence because intra- uterine infection could not definitely be excluded. I say this because later on in this paper I shall instance such cases in which infants were born with the rash well developed. Failing more definite evidence, the period of incubation can therefore be placed provisionally at from ten to fourteen days. Onset. The onset of the disease is sudden. There is a rise of temperature accompanied by headache and backache, and occasionally pains in the limbs and vomiting. The rise of tempera- ture was constant. Of two hundred and two cases of both sexes (one hundred and thirty-three males and sixty-nine females), the incidence of the various symptoms of onset were as follows :— Headache ... ... 172 cases, or about 85 per cent. Backache” =<. Sisley gm TOTS Donte Moi os tg 4 ar Pain in limbs a AN pt ap BH EB 55 Vomiting ... a. Cy LAWTON Oh, — 50 The headache, when present, was generally severe, and often either frontal or vertical. Of the one hundred and eleven cases in which backache occurred, only forty-five described their pain as severe ; the remaining sixty-six described it as moderate. Backache was relatively far more frequent 23 among the women than among the men; 70 per cent. of the former complained as compared with only 45 per cent. of the latter, and twenty-seven of the forty-five severe cases were among the sixty-nine women. The greater incidence of backache among the women is probably due to the fact that many of them were victims of chronic endometritis, and magnified their usual backache symptoms. The combination of headache, vomiting and pain in the back occurred in only twenty-one of the two hundred and two patients, and of these only six vomited more than once, and only one more than three times. In the majority of cases the tongue was furred and constipation was present. Other manifestations. The characteristic eruption appeared with about equal frequency on the third or fourth day after the onset of the symptoms. The actual figures are as follows :— In 21 cases the rash occurred on the 2nd day. In 75 x” a”) ” a) 3rd a”) In 79° ” ” vy a) 4th 3) In 36 ” ” ” a) 5th 29 Either shortly before or after the appearance of the rash, the temperature falls and the constitutional symptoms disappear. The patient is then quite at ease until maturation begins, when for two or three days there is a great deal of pain from the tension under the skin. Jn a number of cases there is also secondary fever. No prodromal rashes were seen. Delirium was never observed. The deep depression which occurs at the onset of true smallpox was uniformly absent. . Menstruation did not appear in the women unless a period was due, and even then no one complained of more than her usual loss of blood. Odour. There was an absence of odour such as is produced by smallpox. A few cases, however, developed a distinctly putrefac- tive smell, which was due to decomposing discharges. Pain in the throat and dysphagia, accompanied in some cases by aphonia and enlargement of the glands of the neck, were noted as occurring in a number of cases. These symptoms were due to the presence of the eruption on the fauces, and presumably in the larynx and trachea. 24 Sputum. Three cases had bronchitic signs in the chest, and for a few days coughed up blood-stained sputum. Bowels. In two cases there was profuse diarrhoea at the onset, but the majority were constipated. Urine. In fifty cases whose urines were examined albuminuria was absent, unless due to some other cause, such as urethral or vaginal discharge. Unfortunately, no urines were obtained before the eruption appeared, and in none of the cases was the examination performed more than once. In one case of diabetes, the eruption ran the usual course, but was followed by a large number of boils. Eruption. Patients do not usually come under observation until the rash 1s well developed ; but in two cases which were admitted to the Isolation Hospital in the pre-eruptive stage the rash appeared in the form of small papules, which to the touch were superficially situated : the papules becoming vesicular in about thirty-six hours. The vesicles are circular in shape, and when fully mature are from 4 to 5 mm. in diameter. The summit is either dome-shaped or flattened, and frequently shows a darkened central area. In the early stages the vesicles, if pricked, yield a clear serum quite free from cells, but polynuclear leucocytes begin to appear in the fluid on the second or third day, and gradually increase in numbers until turbid fluid, or even sometimes thick pus, is formed. At this stage the lesion is very tense, hard and shotty. In the lighter coloured skins a definite red areola surrounds each pock. Primary umbilication is not often, if ever, seen, but on about the eighth or ninth day a secondary umbilication or flattening takes place, and is due to resorption of fluid. The eruption is subject to variation, but, broadly speaking, two main types are distinguishable; the one type being finer and more closely set, and the other being larger and more distinct. Some- times both types are found in the same patient, the vesicles then presenting a very unequal appearance. The finer eruption has far less tendency to.form thick pus, but the general course was similar to that of the larger variety. A number of confluent and two haemorrhagic cases occurred in this series (altogether four cases of haemorrhagic rash have been brought to my notice, all occurring in women six to seven months pregnant, and all fatal). 25 Distribution of the rash. The rash makes its appearance or, at all events, is first noticed in certain positions. These are in order of frequency, the face, especially the forehead, and the dorsum of the wrist or forearm. Of the two hundred and two cases, the location of the onset, as noticed by the patient, was as follows :— Pace~ 5. et oo yeh ee Bt PET 20 Wrist and forearm ... 2 a7 e . 52 Both arm and face ... =~ Mee Ad. 27 Scrotum (2 eB . ae ds wee I Inner side of knee... ae ra oe I Elbow ... ef 2E fh i au I Although in severer cases, as in Plate III, the whole body may be covered, the rash shows a predilection for certain areas. It especially tends to affect the face, the lower half of the back, and the arm and forearm, especially towards the wrists. Scalp. The rash was present on the scalp in all the cases examined ; the lesions, however, were often few in number. Mouth. Pocks were frequently seen on the hard and soft palate, and to a less extent on the pillars of the fauces and the inside of the cheeks. In four cases the fraenum linguae was also affected. Larynx. UHoarseness of voice and sometimes aphonia were present in the majority of the severe cases, and in a fair proportion of the other cases. Laryngoscopy was not possible, but in three of the cases pocks were present in the larynx and trachea—post- mortem. . Palms and soles. In all the two hundred and two cases pocks were seen on the palms and soles. In some these were abundant and caused much pain and discomfort. No lesions under the nails were noticed. Genitalia, especially the prepuce, were often affected, and there was in a few cases much swelling and pain and difficulty of micturition. Plates IV and V show that the rash is present on the area between the knee and the ankle. In true smallpox this area is described as being often free from rash. The parts on which the distribution of the rash is often compara- tively slight are :— 26 Tq dhe neck: 2. The upper part of the trunk, and the abdomen. 3. The inner side of the thighs. 4. The circumorbital area. In this latter situation there is frequently no rash at all, even in severe cases, though pocks are often seen on the edge of the lids. No pocks were seen on the conjunctiva. The effect of irritation appears to be to determine a plentiful outcrop of rash (Plate V). The course. The rash does not appear in crops, but it is often two or three days before the full extent of the eruption is obvious. The order in which it affects the various parts of the body is similar to that of true smallpox. After its appearance it gradually passes through the vesicular stage, already described, until it reaches maturity at about the sixth or seventh day. There is no tense shotty feeling until the rash is nearly matured. This maturation is accompanied by oedema of the subcutaneous tissues. In the majority of cases this oedema is slight, but in others it is so great as sometimes completely to close the eyes. The oedema appears on the third or fourth day of the rash, reaching its height on the seventh or eighth day, and rapidly disappears (Plate V1). Resorption of fluid begins to take place on about the eighth day, and convalescence is so rapid in many cases that by the twelfth day nearly all the scabs have fallen off the face. The rash disappears in the same order in which it appears, and in uncomplicated cases all the scabs have fallen at latest by the end of the third week. In yet cther cases the pigmentation is around the scar, the scar itself being achromic. In yet other cases, in fair skins there has been no subsequent pigmentation. In my opinion, the pigmentation is not of much import, in that the normal negro tends to deposit excess of pigment in and around scars. At first I thought that the pigmentation was the result of local treatment, but changed my mind when I saw the same thing in the scars of two infants born alive after intra-uterine alastrim. The Temperature. The onset of the disease is marked by a rise of temperature, which may reach 104° or even 105°, but in most cases 1s about 103°. This temperature persists with but slight variations for three or four days, then rapidly falls to normal as the rash appears. Sometimes the fall of temperature completely 27 precedes the appearance of the rash, at other times both take place co-incidently. The temperature then remains down for four or five days, to rise again as the rash matures. In mild cases there is no Cuarr I.—Mild case of Alastrim. No secondary fever. Acute onset. DAY oF Ae Af NCCCECCCETT “syed le tl Hin OI Aa) etne aH \cBasesucavaen aay E9Esen GIES ETETETET 2) SHITE: ics NG EP ean al ee secondary rise of temperature, and in the severe cases the rise appears to be in some way dependent on the extent of the vesicles and the amount of infection with skin organisms. There is only a ouge Bie a 28 slight rise when the pus is practically free from organisms, and a greater rise when there are many. In milder cases secondary fever is absent. A curious point about this secondary fever is that the patient is not conscious that he has a temperature, though his temperature may be as much as 102°. After persistence for a few days the temperature returns to normal, and stays there unless complications, such as boils, occur. The secondary fever is as a rule very mild. Cuart III.—Case of Alastrim exhibiting ‘ Typhoid’ type of chart. fest [3 [ie] i) 6 [resliaa aaip [ovaliad oleae re) MESH@PAH SABBRBORBe SL. Peat ne Bena eeee eee Sot 21 ES Hina ROsuhr alia) obeaun: EG : | . ee aE ae ; SFA epee een ese see Bm ea eee ae ei ei There is, however, another type of temperature which has been noted in a few of the very severe cases, and this approximates to the typhoid type, persisting for fourteen or fifteen days before falling to normal. The temperature of onset is no indication of the severity of the disease, high temperatures being often succeeded by a scanty rash. Complications and Sequelae. Broncho-pneumonia is the most serious. It occurred in some of the fatal cases in which much rash was present in mouth and respira- tory tract, and was probably due to aspiration of septic material. Laryngitis and aphonia occur in the severer cases, but disappear as the rash disappears. Conjunctivitis of a mild type develops in a number of cases, and is due to infection from the discharges of lesions on the eyelids. Impetigo. Six cases developed impetigo when the rash was disappearing. In severe cases large areas of skin are apt to be a) stripped off, leaving raw surfaces which are very painful and trouble- some to treat. Boils are the most frequent sequel. They appear at about the fifteenth day after the onset of the rash, and may persist for weeks. Eczema of an intractable character of the external auditory meatus has also been noted. Prognosis is good, except in the newly born and in the haemorr- hagic type of rash. Of two thousand nine hundred and twelve cases which have passed through the Isolation Hospital up to the end of March, 1921, there have been only thirteen deaths, an average of 4°5 per 1,000. In eight of these cases the condition was as follows :— Two women who were 6-7 months pregnant with a haemorrhagic rash. One bled profusely from nose and mouth, vagina and bowel; and both post-mortem showed internal haemorrhages. One man who was 56 years old and died during convalescence. One man who died after admission, but showed no signs of alastrim. One man admitted in a dying condition. He had extensive confluent lesions with skin stripping and leaving large raw surfaces. He could hardly breathe. The mouth was very septic, and the smell from putrefying discharges was very offensive. Three children all within the first month of life and manifesting the disease within the first fortnight of birth. II. OCCURRENCE IN THE FOETUS The virus passes fairly readily through the placenta into the foetal circulation. Up till February, 1921, of twenty cases admitted, after attacks of alastrim, to the Jubilee Maternity Hospital, eight cases of abortion at about the sixth month have occurred, and in each case the macerated foetus was marked with scars of the disease. Two of the cases I was able fully to investigate, and in these the abortion occurred eight weeks after the onset of the disease in the mother. All the organs were searched for spirochaetes without result, and the Wassermann reaction of the blood of the mothers was negative. The scars in the foetus were slightly depressed. In addition, two children were born alive with marks of alastrim. The first child was born at full term with marks present as follows :— 30 Hace. ae Lith aoe Me 9 marks. Trunk (front) ... 26 st 52 alO hess Trunk (back) ... wth s8 rh. deoare oils 3 A few on each leg and arm. Two on each sole and one on each palm. These scars were depressed and surrounded by pigmentation. The mother of this child was alone in the world and developed eclampsia, and died soon after the birth of the child, so that it was impossible to obtain an accurate history. Scars and pigmentation such as occur after alastrim were, however, present on her body. The second child was born at the seventh month and had no sign of disease on the face, but six spots on the left arm and seven on the right, with three on each leg. These scars were pigmented, as is the case in adults. The mother’s attack occurred eight and a half weeks previous to the birth of the child. The mother's Wassermann reaction was negative. In none of these cases of foetal alastrim was the disease very severe in the mother, judging by the amount of scarring and pigmen- tation present. The remaining ten labours yielded normal, full term children, two of which developed alastrim a day-after labour. In addition, two remarkable cases have occurred in which mothers who had been vaccinated, who have never had alastrim, gave birth to children covered with an alastrim rash. Both cases present a very similar history, save that one mother was vaccinated six weeks and the other four weeks before labour. I give the details of the second case. *P.C.,’ age 23. Sailed from Cuba, January roth, for Jamaica ; was vaccinated on the day of sailing. She landed in Jamaica January 12th, and had fever on 14th and 15h January. So far as she is aware she has never come into contact with any active cases of alastrim. On February roth (day of examination) the scab had not yet fallen off her vaccination mark and covered an area a little larger than that of a threepenny-bit. She had no signs either in the way of scars or pigmentation of having had alastrim. The child was born at full term with a pustular eruption (Plate VII), and died five days after birth. Mother and child gave negative Wasserman reactions. These are cases either of generalised vaccinia occurring in utero 31 or of alastrim transmitted to the foetus by a mother rendered immune by vaccination. If they are cases of generalised vaccinia they demonstrate that ordinary vaccination can be so transmitted ; if they are cases of alastrim, it would appear that just as the diphtheria bacillus grows readily in diphtheria antitoxin so the organism of alastrim can flourish in the blood of one who has, by vaccination, been rendered immune to its toxin, can retain its virulence, pass through the placenta and affect the foetus. There is finally the possibility that the disease may have existed in the mother, but was so mild as to have been completely over- looked even by herself. If the incubation period be regarded as twelve days, and if the mother were infected on the twelfth day, the day of landing in Jamaica, she ought to have manifested symptoms on January 24th, at a time when, on general principles, she would have been completely protected by vaccination. The child at birth had a rash of at least five days’ duration, and if another three days are allowed before the rash appears, must have been manifesting symptoms in utero by the 2nd February. If the child were infected twelve days previous to the manifestations of symptoms, the time relations would be about right. But as against this conclusion, the mother maintains that, apart from slight fever on the fourth and fifth days after vaccination, she was perfectly well. Moreover, I have not seen a mild case of alastrim in which there has not been some malaise. Of the three mildest cases which came under my notice one had four and two had two pocks each, yet in each of these cases the eruption, such as it was, was preceded by fever and malaise. Therefore, pending further evidence, I am of opinion that these cases illustrate the possibility of an immune mother transmitting the disease to her unborn child. In connection with the question of foetal alastrim, I am impressed by the relatively high frequency with which it occurred among the cases of labour admitted to the Jubilee Hospital. Of twenty cases, ten produced alastrim foetuses, two produced infants developing the disease one day after birth, and only eight produced normal infants. 32 III. ITS RELATION TO VACCINATION The following two tables summarize the facts in two hundred and six adults and eighty children taken at random :— ADULTS (206) Cases | Mild Medium Severe pa yi a ee Yo % % With vaccination scars... oo ae 72 | 47°2 30°5 22'°2 Without vaccination scars aes sos] 134 28°3 3453 37°3 | Total “ss ee 206 3570 33°0 32°0 CHILDREN (80) Cases | Mild Medium Severe 0, of of 0 /0 oO With vaccination scars... 288 ace 26 | 84°6 15°4 o"o Without vaccination scars = ao ca) | 44°4 | 25°9 209°6 | 23°0 20°0 Total ate se) 80 570 It will thus be seen that : — 1. There is a tendency to a mild type of case in children, due partly to vaccination and partly to some other factor. 2. No severe case occurred in the vaccinated children of this series, and twenty-two of the twenty-six (84°6 per cent.) vaccinated were mild cases. Professor MacCallum and myself were continually exposed to infection for hours at a time, but never contracted the disease. We were both vaccinated, he recently and myself five years ago. One of the helpers at the Isolation Hospital was vaccinated by me before taking up duty. I frequently watched her handle the patients and 33 then either put her unwashed hands to her mouth, or wipe them in a handkerchief which she subsequently used to wipe her face. She never manifested any symptoms. The Medical Officer of Health, Kingston, has vaccinated more than five hundred contacts, and he states that no cases of alastrim have occurred amongst them. I have vaccinated .twenty contacts, and these were also completely protected. On the other hand, five cases occurred in vaccinated infants, three in infants two years old, and two in infants four years old. Goldsmith and Loughnan (i921) give clinical notes of three cases occurring in the vaccinated, three years, one and a half years and one year after successful vaccination. Re-vaccination after alastrim. Sixty cases were vaccinated by me during convalescence, the time of vaccination varying from the fifteenth day after the onset of the rash up to the twelfth week. O} these, fifteen showed no sign of ‘take’ in two weeks, but twelve of these fifteen were subsequently vaccinated with three ‘takes.’ The method used was that of simple incision through the surface layers of the skin. Of the forty-five ‘takes’ none was typical as compared with normal controls vaccinated with the same batches of lymph. Course. The incision healed over, and no sign of ‘take’ was visible before the seventh or eighth day, when a few papules were seen in the line of the incision. The papules increased but slowly in size, and by the fourteenth day were raised about 2 mm. above the surface of the arm. No induration or swelling of the arm was noticed, and in only a few cases was there adenitis, temperature, malaise or an areola more extensive than }-c.m. around the lesion, and even in these few cases the symptoms were hardly noticed. On pricking the early vesicles a small quantity of clear fluid was obtained. The lesion was multilocular, and on removing the top of the vesicles in a few cases it was seen that the base was composed of exuberant granulations rising up above the level of the skin, accounting for the fact that sometimes on pricking a small amount of blood came out with the fluid. ~ The scabs fell off in four weeks or more, and with one exception the. resulting scar was not the depressed, pitted scar of typical 34 vaccinia, but on the contrary slightly raised (hypertrophy) above the surface, subsequently contracting to the level of the surface. The points of difference between this and typical vaccinia appear to be: — 1. Tardy development and course of lesion. 2. Small size of vesicles and exuberant base and imperfect umbilication. 3. Insignificance of both local and constitutional reaction. 4. Resultant scar. The evidence of the nature of vaccination after alastrim seem to show that vaccine lymph contains something else beside the factor which protects against alastrim, and it is this something else which gives the reaction after alastrim. The vaccine lymph on examination was found to contain a large amount of Staphylococcus aureus, an organism which is present in many specimens of calf lymph, and it may be this organism which causes the reaction, but the lesion produced did not suggest a septic process. I inoculated four rabbits with the fluid from the vaccine lesion of one of the post alastrim cases, but the results were negative. My opinion is that alastrim and vaccinia belong to the same group but present slight individual differences, the one disease affording almost complete immunity to the other. IV. MORBID ANATOMY Case 1. Post-mortem performed 16 hours after death. Death took place on the 12th day of disease. Patient thickly covered with rash, which was confluent in parts and presented inequality of size of vesicles. Small petechial haemorrhage was seen on the sides of the abdomen. Patient was 6 months pregnant. She was admitted bleeding from nose, mouth, uterus. The mouth and throat were filled with a mass of necrotic material, and the entrance to the larynx and the vocal cords was similarly covered with necrotic material. The trachea contained pocks in its entire length. The right lung showed a haemorrhagic condition. There was a broncho-pneumonia with small spots of scattered haemorrhage visible on the cut surface of the lung. The left lung was apparently affected in the same manner, but to a less extent. The heart showed petechial haemorrhages under the epicardium, especially at the root of the aorta, and gross haemorrhage into the muscle substance of the left ventricle. Valves normal. The kidneys presented gross haemorrhage in the medulla and the pelvis was filled with blood. The bladder also showed haemorrhage under the mucous membrane, especially around the outlet. The liver was enlarged, but to the naked eye not obviously abnormal. Spleen not enlarged, firm. Stomach filled with ‘coffee grounds’ material, 35 petechiae present under mucous membrane. Intestines: colour dark red, with much haemorrhage and oedema. This condition affected the duodenum and first 3 feet of the jejunum. No lesion seen in the foetus. Case 2. Post-mortem six hours after death. Patient well covered with rash, which, on the face, was beginning to crust. Some confluence present on left side of abdomen. Patient 7 months pregnant. The throat was filled with necrotic material as also was the entrance to the larynx. Trachea showed the presence of pocks in its entire extent, and a similar condition obtained in the bronchi. Right lung showed some small haemorrhages and broncho-pneumonia. Left lung apparently normal. No haemorrhage into pleura or pericardium. Heart: A few spots of haemorrhage over the right ventricle under the epicardium. No gross haemorrhage into the substance of the muscle. Valves normal. Liver much enlarged and fatty. Spleen normal. Kidneys normal in size, but pale and fatty. Capsule stripped easily. No gross haemorrhage seen. Bladder normal. Uterus : A large subperitoneal haemorrhage on the right side just below the fallopian tube. Uterus contained a 7 months’ foetus, which, on examination, showed a few sub- epicardial haemorrhages. Stomach normal. Intestines: The first 2 feet of the jejunum presented a remarkable condition; they were dark, haemorrhagic, and very oedematous. The whole lumen was in parts a solid mass. Caszt 3. Death on 11th day of rash. Scabbing on face and in upper part of chest. There were areas on the arms, abdomen, and particularly on the legs, in which absorption of the fluid in the vesicles was taking place leaving the vesicles lax. There were also large areas of confluent eruption about the size of half-a- crown on the outer portion of both legs, and on the sides of the abdomen. Some of the fluid from these areas was blood-stained. On other parts of the skin where the rash had been confluent the epidermis was stripping off leaving the surface raw. A putrid odour was observed. The trachea had small, ulcerated lesions extending down to the bronchi. The lungs were apparently normal except for a small portion of the upper lobe of the right lung. This appeared to be con- solidated. Heart firmly contracted with somewhat excessive fat under the epicardium, otherwise normal. Aorta: Some athero-sclerosis. Liver large, somewhat congested, and on section looked like a nutmeg liver. Spleen smaller than normal, firm and fibrous. Kidney normal in size. Capsule stripped easily, substance rather pale, vessels more prominent than normal. Suprarenals normal. Bladder normal. Uterus: 2 small subserous fibroids. Intestines normal, but filled with a very large amount of faeces, especially the sigmoid and rectum. Cas 4. Infant born with scabbing eruption; died 4 days after birth. General appearance normal save for eruption. Mouth, larynx, trachea normal. Lungs presented an unusual appearance; they were reddish-brown in colour, and on the surface was scattered small, white areas about 2 mm. in diameter. These areas extended for a short distance into the surface of the lung. The lungs were partially solid. Liver was dark red in colour and presented an appearance similar to that of the lung. Spleen apparently normal. Kidneys: There were small haemorrhages under the capsules and extending into the cortex. Suprarenals normal. Heart and circulatory system normal. Cast 5. Infant born with alastrim. Mouth showed rash on the inside of the cheek. Larynx and trachea normal. Lungs and liver: condition similar to those of Case 4. Spleen and other organs normal. Inoculations. Fluid was collected from the skin lesions of patients in various stages of the disease, and twenty-six rabbits and 36 four calves were inoculated by Professor MacCallum and myself. The skin was shaved and scarified, in some cases drawing blood, and the material was well rubbed in. These all gave negative results. Blood from early cases and from a few patients who after contact with cases had developed headache and fever, but none of whom subsequently had an alastrim rash, was also used, but with negative results. REFERENCE Go.psmitH and LouGunan (1921). Fourn. R.A.M.C., Vol. XXXVI, p. 66. 4 4 ts yaly iipited) “bolnnia oe “slhitentiige ie dew pab ats aly KSI Doms ca S iw - sie al) to ewiae SwilAtiaoey iT « © hh en epi (3) IS 38 EXPLANATION OF PLATE III Severe case. Patient vaccinated twelfth day after exposure to infection. Rash developed on eighteenth day and ran concurrently with vaccination. Some of the lesions show secondary umbilication due to resorption of fluid. Annals Trop. Med. S Parasitol., Vol. XVI PLATE Ill — 7 gi MOTAVS siege : Bids bic peri kyt 40 EXPLANATION OF PLATE IV Showing distribution of rash in a moderate case. Annals Trop. Med. & Parasitol., Vat. XVI { PEAGE LF C. Tinling & Co., Lid., Imp. 42 EXPLANATION OF PLATE V Showing effect of irritation. Note patches on inner side of left thigh above knee. This crop came out around a septic cut. Annals Trop. Med. & Parasitol., Vol. XVI BEATE C. Tinling & Co., Lid., Imp. AVA 8 eet) 6 We ae ar > } ; <8 ibs bent 7 nit ake Ww a] ries 7 -* ee) , ba x - 44 EXPLANATION OF PLATE VI Eighth day of rash. Left eye almost closed. Annals Trop. Med. & Parasitol., Vol. XVI : 12) ay: i a Da La Gre Tinling & Co., Lid., Imp. i , a _— 1} — Pei : ; r : rf mY res . el * s >t < : sp © a ; : b ' ’ , ” FAG AO MOL eA es iy id a= a a a _ — - ‘ Agen aks rls lige iiw die) Tin? ie cod iid’) ioleony inet Cin: beetanieos a s- ‘a eal tae i wits AOS yer Iilivin f 46 EXPLANATION (OF “PEATE Vit Child born at full term with well developed rash. Mother vaccinated one month previous to birth of child. No history of illness in mother. Wassermann reactions of mother and child negative. Annals Trop. Med. & Parasitol., Vol. XVI PEAR lL C. Tinling & Co., Lid., Imp. 47 A NEW SPECIES OF PHLEBOTOMUS FROM TRINIDAD BY Prof. R. NEWSTEAD, F.R.S. (Received for publication 23 January, 1922) Phlebotomus trinidadensis, sp. n. A relatively small species. CG genital armature with five large spines to the superior claspers: three terminal or distal, and two slightly beyond the middle distance, arranged with their bases on opposite sides of the segment; no tufts of non-deciduous hairs, proximally. Colour of both sexes similar. Pale ochraceous. Wings with the costa sometimes distinctly infuscated, and in certain lights with intense iridescent blue. Legs silvery grey. Male. Abdominal hairs of the medio-dorsal line arranged in small, sparse groups on all of the segments ; those of the venter dense, some of them semi-erect, others procumbent. Hairs of the proximal segments of the superior claspers very long and dense. Palfz of five segments: second, third, and fourth equal in length; the third and fourth broadened distally ; fifth, two and a half times longer than the fourth. Axdennae with the third segment projecting slightly beyond the tip of the proboscis; geniculated spines relatively very small, and apparently bilateral, those on the third about one-eighth of the entire length of the segment; those on the sixth and seventh a little less than one-fourth the entire length of the segments respectively. Wzxgs (fig. 1 @) moderately narrow, the fork of the fourth vein generally in advance of the proximal fork of the second. Genital armature (fig. 1 6), relatively large; superior claspers each with five long, stout spines: three distal and two slightly beyond the middle distance, the latter arranged with their bases on opposite sides of the segment, the two outer, distal ones and the inner, O:? mm. Fic. 1. Phlebotomus trinidadensts, sp.n. 3: a, wing; 4, genital armature. Q: c, wing; d, terminal segments with the paired leaf-like appendages. a, ¢ and d to same magnification. 49 lateral one longer than the others; the middle, distal one shortest ; the segment about half the length of the proximal one. Inferior claspers slightly shorter than the proximal segment of the superior claspers. Length 2°3 mm.; length from front of thorax to end of armature 1'8 mm.; wing, 1°3 mm.; leg III, 2°6 mm.; internal genital armature, o'6 mm. Female. More robust and generally larger than the 5. Addominal hairs more or less erect. Palpi similar in form to those of the ¢. Third segment of antennae shorter than the corresponding segment in the g, and not reaching the tip of the proboscis. Wings (fig. 1c) much more broadly lanceolate than in the d ; curvature of the borders similar; venation similar to that of the @. External genitalia (fig. 1d): the superior leaf-like appendages relatively exceptionally large and, in macerated specimens, widely separated from the inferior pair; both appendages strongly hairy; the inferior pair with the finer hairs on the distal two thirds arranged in distinct, equidistant rows. Length, 2°1 to 2.9 mm.; length to front of thorax, 1°7 mm.; wing, 1°6 mm.; leg III, 3:1 mm. Two American species: P. vexator, Coquillet (1907) and P. brumpti, Larousse (1920), resemble this species in regard to the number of spines on the superior claspers. In db7amfti, however, the armature generally resembles that of P. papatasii, and is, therefore, markedly distinct. P. vexator also differs in having the spines arranged as follows: two apical, ¢wo sub-apical, and one in the middle of the segment. In P. ¢rinidadensis, sp. n., the formula is three apical and ¢wo near the middle of the segment. TRINIDAD. Six 3d, seven 9 ? (four of which contained blood), 1921. Major W. F. M. Loughnan, R.A.M.C., D.A., D.P., West Indian Command, with the assistance of Captain D. A. MacDougall, M.C., R.A.M.C. In his letter from Kingston, Jamaica, dated 22nd August, 1921, Major Loughnan states that he, together with his fellow-officer, had a good deal of trouble in finding the specimens, and further that the species appeared to be very sparse in its distribution in the Island of Trinidad. This is the first authentic record of the occurrence of a species SO of Phlebotomus from the West Indies; and the captors are to be congratulated on their interesting discovery. Possibly other new and undescribed species await the hunters of these small midges in that region. REFERENCES Cogquittet, D. W. (1907). Entomological News, Vol. XVIII, p. 102. Laroussk, F. (1920). Bull. Soc. Path. Exot. Vol. XIII, pp. 659, 662. gi A NEW TSETSE-FLY FROM THE SOUTH CAMEROONS BY Professor R. NEWSTEAD, F.R.S. AND Miss ALWEN M. EVANS, M.Sc. (Received for publication 10 February, 1922) In the course of his investigations in the South Cameroons, during the past year, Dr. J. Hanington, a former member of the Staff of this School, made several small collections of tsetse-flies and other blood-sucking Arthropods. These he has generously presented to this Institution for the Museum collections. In the last consign- ment, which reached us towards the end of January of this year, were many examples of Glossina palpalis, R. D., G. pallicera, Bigot, and four specimens of a large species which, on microscopical examination of the morphological characters of the genital armatures, proved to be new and undescribed. In the letter accompanying the collection of flies, Dr. Hanington submitted a sketch-map of the districts through which he had passed, and gave the following brief account of the nature of the country in which the captures were made: ‘The tsetse-flies were collected on my just completed tour of inspection N.W. over our border to Obudu. The country is hilly, forested, with many swift, shallow rivers, and full of tsetse. The large species is found only in the neighbourhood of Basho, where the ground begins to rise to the north into fly-free mountain-plateaux. The greatest number of tsetses were on the Mbilesi-Mateni Road, which runs along a wooded and rocky river valley.’ The commonest species in this region would appear to be Glossina pallicera, of which twenty-six specimens were sent. We append a description of the new species, and have ventured to dedicate it to Dr. Hanington, the discoverer, in recognition of his devotion to the science of tropical medicine. 52 Glossina haningtoni, sp. n. A large dark-coloured species, with infuscated wings, belonging to the ‘Fusca Group. Hairs of the third antennal segment relatively short. Proboscts (palpi) 0-7 to o'9 mm. shorter than in G. FuscA. Waedth of front in both sexes similar. Harpes of male each with three processes, the distal one angular and emarginate in front. Signum of female with height slightly exceeding width and paired crescentic folds almost continuous behind. Male: Length, 11 mm.; proboscis, 4 mm.; front of head, 075 mm.; wing, 11 mm. Female: Length, 11 to 12 mm.; pro- boscis, 4°2 mm. ; front of head, 0°75 mm. ; wing, 12 mm. Male: Head with the posterior surface ‘mouse-grey’ (Austen), with a narrow black streak on the upper surface bordering the narrowly pale margin of the eyes. Vertex immediately behind the ocelli with a narrow black area. Front pale brown with a much paler area surrounding the ocelli. Antennal cavity greyish below, sides a little paler than the front. Ax¢ennae with the first two segments dark brown; the third pearly-grey, the tip of the segment moderately prominent, with the outstanding hairs forming the fringe in front from one-seventh to one-eighth the width of the segment. Pvoboscis relatively short, bulb uniformly pale buff- yellow. Thoracic markings very dark and pronounced, suture and ground colour forming the trident-lke marking immediately in advance of the scutellum, pale ochraceous, the rest darker. Abdomen: Dorsum of first and second segment brown; the rest very dark, glossy sepia-brown, distal angles of last three segments ochraceous-grey ; venter orange-ochraceous. Legs orange- ochraceous: leg I, with the femur infuscated along the dorsal half, tibia infuscated externally, tips of last two segments of tarsus dark brown or black; leg II, similar to the first but lhghter in colour; leg III, with the third and fourth segments of the tarsi dusky, the last two all dark brown or blackish; all the ventral hairs dark golden. JWexgs rather strongly infuscated. Genital armature (fig. 1): Harpes (4.) with three bi-lateral processes; proximal pair long and spine-like, the first slightly shorter than the second; distal process angular, and when flattened by pressure shows a deep emargination on the distal margin (#.1), but with the lower, 5S angular projection folded inwards the emargination almost entirely disappears (4. 2). Ventral chitinous sclerites long and projecting almost as far as the distal processes of the harpes. Inferior claspers (z. c.) normal, a few of the marginal hairs of great length. Median process with its distal edge rounded, and projecting slightly beyond the inferior claspers. Superior claspers (s. ¢.) relatively rather long, and as usual, bluntly bifid. Fic. 1. Glossina haningtoni, Newstead and Evans. ¢ genital armature: s.c., superior claspers ; i.c., inferior claspers; i.c./., two hairs from the inferior clasper, one of them malformed ; b, harpes ; 4.1, distal process of harpe, with lower arm extended, internal aspect ; 4.2, the same with the lower arm curved inwards, external aspect. Female: Third antennal segment pale ochraceous proximally, the distal three-fourths infuscated. Colour of legs, abdomen and plurae slightly darker than in the male. The ‘ black streak’ on the posterior surface of the head absent. The space between the eyes (front) as inthe male. Genital armature: External armature of the type found in Glossina fusca but the dorsal plates rather broad, the 54 width exceeding one-third of the length. Internal armature, signum of uterus (fig. 2) measuring 0°41 mm. in height and 038 mm. in greatest width. Median portion of signum (#7. ~.) a thin plate of the form shown in the figure, ochraceous brown behind, becoming straw-coloured towards the anterior margin; postero-lateral portions (p~. l. p.) laminar, pale ochraceous, connected with the median plate by deep crescentic folds of black chitin (c.c.) These folds almost continuous posteriorly, and forming a striking feature of the signum. SOUTH CAMEROONS: Basho, Mamfe (Ossidinge) Division, 14th December, 1921, 2 99;2 66. Dr. J. Hanington. ee A.M.E. Fic. 2. Glossina haningtont, Newstead and Evans. Q Signum: c.c., crescentic fold; m.p., median plate; p./.p., postero-lateral plate. Closely related to Glossina fusca, but differing externally by the relatively much shorter palpi (proboscis), and the slightly more robust appearance. But the most marked morphological differences can be seen only in the genital armature of both sexes. A careful study of these organs at once reveals the strikingly distinctive features of this species, and its affinities with other members of the ‘Fusca Group’ of tsetse-flies. 55 NOTES ON AUSTRALIAN CESTODES BY P. A. MAPLESTONE (Received for publication 10 February, 1922) III. COTUGNIA OLIGORCHIS, n. sp. On four occasions specimens of the cestode about to be described were found in the intestine of the Whistling Duck (Dendrocygna arcuata, Cuvier), shot a few miles from Townsville, North Queensland. EXTERNAL ANATOMY. The largest specimen measured 80 mm. long and 8 mm. broad at its widest part; these dimensions were taken from fixed material. The scolex is relatively small and there is no neck. In well fixed specimens the worm is of almost uniform breadth for the greater part of its length, but tapers fairly rapidly and evenly both anteriorly and posteriorly. The posterior end is not unlike the anterior, except that it is not so finely pointed, owing to the absence of a scolex. Head. The scolices were not well fixed, being in all cases more or less shrunken, so the detailed characters of this structure cannot be accurately given. However, it is seen to bear a very small retractile rostellum armed with a single row of minute hooks measuring about 10m long, but unfortunately their exact number could not be determined, because all dvailable specimens were imperfect. The four small suckers are situated quite near the anterior extremity, and measure about 0°65, in diameter (fig. 1). Segments. The proglottides are from first to last much broader than long. INTERNAL ANATOMY. Muscular system. On examining transverse sections it is seen that the muscle layers are disposed in the same way as in Diploposthe laevis (Bloch, 1872); Jacobi, 1896. That is, there are a few diagonal fibres externally, with a layer of transverse fibres internal to them. Next in order from without inwards is the main longitudinal layer, which consists of a large number of closely set 56 bundles oval in cross section, with their long diameters running dorso-ventrally, and about o'50m in thickness. This layer is evenly developed and encircles the segment, except where it is pierced by ae ao ET 6 AM.B. del. Fic. 1. C. oligorchis, n. sp. Scolex and anterior portion of strobila. we wa AM B. eel eXyv. Fic. 2. C. oligorchis, n.sp. Mature segments. ., cirrus; ¢.p., cirrus pouch ; ex.v., excretory vessels; ov., ovary; #., testes; ut., uterus; v., vagina; v.g., vitelline gland; v.s.. vesicula seminalis. % 20. a cirrus pouch. Internal to this is another thinner layer of trans- verse muscle with a few scattered bundles of longitudinal muscle irregularly placed on the dorsal and ventral surfaces respectively, and about fifteen to twenty in number on each surface. Internal to these bundles are a few fibres of transverse muscle. It should be noted that the transverse muscle layers consist of hoop-like strands of fibres discontinuous with each other antero-posteriorly, so that in transverse sections they are only seen here and there. (Figs. 3 57 and 4 do not show the transverse fibres for this reason.) The dorso- ventral fibres are most marked in sections through the anterior and posterior of a segment. Nervous system. The nervous system is poorly developed and consists of a small main nerve lying well to the outer side of the excretory canals, and ventral to the cirrus pouch and vagina. Excretory system. The two lateral excretory canals on each side lie in the anterior portion at some distance from each other; the smaller dorsal vessel lies to the inner side of and dorsal to the ventral vessel. In this part of the worm the dorsal canal pursues a dm. | m. t Fic. 3. C. oligorchis, n.sp. Transverse section through a mature segment. c.p., cirrus pouch; d.m., diagonal muscle; ex.v., excretory vessels; 1.m., longitudinal muscle ; ov., ovary; ¢., testes; ut. uterus. X 20. SOS RE EO ACT Vers cp. eXN. cp Fic. 4. C. oligorchis, n.sp. Transverse section through a gravid segment. ¢.p-, cirrus pouch; ex.v., excretory vessels; w., uterus. 20. wavy course, each wave extending over two or three segments, so that the width of the medulla varies slightly in different segments (fig. 1). More posteriorly it straightens out like the ventral vessel and pursues a direct antero-posterior course, and at the same time comes to lie close to the inner side of the latter. Throughout their whole length the canals are a considerable distance from the lateral borders of the worm. Genitalia. The male and female organs lie in the medulla in two separate groups, one on each side of the mid-line. Testes. The testes vary from three to five in number on each side and are situated posterior to the ovary, close to the posterior border of the segment in a transverse line. They measure about 58 Sou in diameter, but this dimension is only approximate, for when five are present they are smaller than when only three are present on each side. Their number and position in relation to the ovary (whether lateral or mesial to it) vary-in different proglottides of the same chain, or even on the two sides of the same segment. Thus when there are three testes they may all lie external to the ovary, or there may be one internal and two external; if four in number they may lie, two internal and two external, or three external and one internal; and lastly, if five in number they may lie three external and two internal, or four external and one internal (fig. 2). They come to full development far in advance of the ovary, and are beginning to atrophy before this organ is fully developed. Vas deferens. There is a small but distinct vesicula seminalis, which lies just internal to or overlapping the excretory canals; from its outer side a narrow, lightly coiled tube leads to the base of the cirrus pouch which it enters. It runs on the dorsal side of the excretory canals. The cirrus pouches are long and relatively thick saccular organs lying transversely near the anterior margins of the segments. In the early stages of development, the cirrus pouches on each side lie to the inner side of the excretory canals, but they soon pass to their outer sides, which relation they then maintain to the end of the chain. They are about 6304 long and rI1o0yu broad, and open into small chambers which in turn open in distinct pores situated on the lateral borders, not far from the anterior lateral angles. The cirri are often seen partly extruded through these pores, and they are relatively thick and straight, being of the same diameter for their whole length, with slightly rounded tips. They are about 460u long and 45 in cross section, and their outer surfaces are thickly covered from base to tip with small straight spines set perpendicular to the surface and about 7# long. These organs are the same in appearance and have the same relations with the other organs on both sides of each segment (fig. 2). Ovary. The paired ovaries are large and are situated one on each side of the mid-line, about mid-way between the anterior and posterior borders of the segments. Each consists of four or five lobes, which radiate forwards and laterally from a central point; the small compact vitellarium lies close behind them. The distances of the ovaries from one another, and consequently from the lateral 59 borders on the corresponding sides, vary slightly in different segments. The shell gland, as a rule, is not clear; but in some segments it can be seen lying between the ovary and vitelline glands (fig. 2). Receptaculum and vagina. The vagina is a relatively wide tube; running from the ovary it first curves forwards and outwards, then turns and runs directly outwards, and crossing ventral to the seminal vesicle, but dorsal to the nerve and excretory canals, it finally runs ventral to the cirrus pouch to open at the genital pore on the ventral side of this organ. The final part of its course can only. be determined in sections. Uterus. The uterus is visible at an early stage as a thin trans- verse tube crossing the proglottis almost from one side to the other, about mid-way between the anterior and posterior borders of the segment (fig. 2). As it develops, it throws out numerous branches in every direction, which gradually increase in size, so that eventually the uterus appears as a broad saccular organ occupying nearly the whole of the segment, with a few trabeculae representing AMBedet Fic. 5. ©. oligorchis, n.sp. Uterus in intermediate stage of development. ¢.p. cirrus pouch. X 20. AMB. del. Fic. 6. C, oligorchis, n. sp. Fully developed uterus. c¢.p., cirrus pouch. X 17°5. the remains of the original branches (figs. 5 and 6). When the uterus is fully developed, the eggs lie singly in capsules. In sections showing the early stages of the uterus, it is seen to pass between the testes (dorsal) and the ovaries (ventral), and at the sides it crosses the excretory canals dorsally and runs almost to the edge of the segments (fig. 3). Later, it pushes the canals ventrally, and tends somewhat to disturb the contour of the muscle layers 60 (fig. 4). No fully developed eggs were seen, the most mature ones measured about 43 and the oncosphere 26 in diameter. DIAGNOSIS. Up to the present, ten species of the genus Cotuguia have been recorded (Meggitt, 1920). All except C. d7ownz, Smith, possess numerous testes. C. d7owni has six to seven testes, but these lie anterior to the female glands. The present species possesses only from three to five testes on each side, and these lie posterior to the female glands; it is thus obviously new, and is accordingly named Cotugnia oligorchis on account of the few testes. The type specimens are in the museum of the Liverpool School of Tropical Medicine. NotEe.—Dizfploposthe laevis, Bloch, was first recorded in Australia by Krefft under the name Zaenia tuberculata; this material was re-examined by Johnston (1912), who assigned it to the above species. The host, in this case, was Aythya australis, Gould, the White-eyed Duck or Widgeon. Later on, Johnston (1913) recorded the same cestode in Queensland ; this time the host was Dendrocygna arcuata, Cuvier, and his specimens came from the Australian Institute of Tropical Medicine. The writer, on examining the slide of this cestode, placed in the Institute museum by Johnston, found that beyond doubt it is a worm of the above described species with two ovaries, and is not D. laevis. Therefore the record by Johnston of D. laevis in the host D. arcuata is not correct. However, D. laevis does exist in Queensland, for the writer has recently examined some material at the Australian Institute which proved to be D. laevis; these worms were taken from A. australzs, the original host in which Krefft found it in New South Wales. REFERENCES Jounston, T. Harvey (1912). A Re-examination of the types of Krefft’s Species of Cestodes. Records of the Aust. Mus. Vol. IX, No. 1, p. 4. (1913). Cestodaand Acanthocephala. Aust. Inst. of Trop. Med. Report for 1911, p. 91. Mecerrt, F. S. (1920). A Contribution to our knowledge of the Tapeworms of Poultry. Parasitology. Vol. XII, No. 3, p. 306. 61 NOTES ON AUSTRALIAN CESTODES BY Pp. A.) MAPLESTONE AND T. SOUTHWELL (Receved for publication 10 February, 1922) IV. GYROCOELIA AUSTRALIENSIS, Johnston This cestode is evidently fairly common, as it was found in the intestines of several members of the species Spur-winged Plover (Lodivanellus lobatus, Lath.) shot in the neighbourhood of Townsville, North Queensland. EXTERNAL ANATOMY. Fixed worms measured 167 mm. in length, with a maximum breadth of 4 mm. The worm is very narrow anteriorly, and widens fairly rapidly and evenly posteriorly. The most striking character is its dorso- ventral diameter, which is very great, especially towards the posterior end. The segments are thick in the centre and thin at the edges, so that in cross section they are bi-convex. The large regularly alternating cirrus, extruded in most mature segments, can be easily made out with the naked eye. Head. The scolex is flat anteriorly, and measures 0°315 mm. in breadth and 0°22 mm. in length. From the centre of the anterior surface arises a thin rostellum about 1204 long and 40, broad, tapering anteriorly and ending in a bluntly rounded tip; there is very little muscle in this organ. Unfortunately, in all our specimens the hooks had been lost. The four suckers are placed, two on the dorsal and two on the ventral surface of the scolex, and look directly dorsally and ventrally respectively. They are circular in outline, and measure about 130” in diameter. Behind the scolex there is no true neck, but a short unsegmented portion of about the same width (fig. 1). 62 Segments. Segmentation begins at a distance of 4°8 mm. from the anterior end, and about the first ten segments become successively narrower, thus giving rise to the appearance of a neck. The minimum breadth is about 1704; from this point the segments progressively increase in width to the posterior extremity. The dimensions of mature segments are 1°25 mm. across the anterior, and 1°5 mm. across the posterior borders, with a length of 08 mm. It is thus apparent that the posterior angles are slightly projecting (figs. 2 and 3). Fic. 1. G. australiensis. Scolex and anterior portion of strobila. xX 56. INTERNAL ANATOMY. Muscular system. In transverse sections the relatively great thickness of the cestode is seen to be due chiefly to the longitudinal muscle fibres, which are arranged in two distinct layers (fig. 4). From without inwards the structures are arranged as follows. Furst there is the cuticle, which is about go# thick, then a layer of transverse muscle, and next to it the outer layer of longitudinal 63 muscle. This measures about 60 in thickness, and is composed of oval, discrete bundles of muscle fibre lying with the long axis of the bundles dorso ventral. On the inner surface of this layer is another thin band of transverse muscle, which has on its inner surface the A.M. B. del. AMB. del. Fig. 2. G. australiensis. Young segments Fic. 3. G. australiensis. Older segments showing showing male genitalia, c., cirrus; #., testes; female genitalia. c., cirrus; ov., ovary; v.g., vitelline v.S. vesicula seminalis. X 35. glands. X 35. tm1.tm2.tm3. esos Taasdel 4 1 Bee Lm.1. 1m2° eX (e- AMB det Fic. 4. G. australiensis. Transverse section of mature segment. ¢., cirrus ; ex... excretory vessels; l.m.1, outer layer of longitudinal muscle; /.m.2, inner layer of longitudinal muscle; ov., ovary; ¢.m.1, outer Jayer of transverse muscle; ¢.m.2, middle layer of transverse muscle ; #.m.3, inner layer of transverse muscle. X 20. second and thicker layer of longitudinal muscle. The dorso-ventral diameter is about 140” in the mid-line, and gradually decreases towards the sides; like the outer layer it is composed of oval bundles of fibres, but in many cases these are broken up into smaller 64 subsidiary bundles. Dorso-ventral fibres can be made out running between the bundles of longitudinal muscle. On the inner, side of the second layer of longitudinal muscle are a few scattered trans- verse fibres, and these with the two outer layers of the same fibres end in the outer coat of the cirrus pouch where this organ is present. Nervous system. The main longitudinal nerve is situated well to the outer side of the ventral vessel, and ventral to the cirrus pouch. Further details of this system were not investigated. Excretory system. The dorsal-lateral excretory vessel is smaller in diameter than the ventral, and lies directly dorsal and close to it, except when the cirrus pouch passes between them, where they become more widely separated. Testes. The testes are found only in young worms in which there is no trace of the female genitalia, even in the terminal segments. They first appear about the thirtieth segment, and from this point posteriorly they gradually become more developed, and again dwindle, until at about the hundredth segment they have quite disappeared. The testes number about fifty in full develop- ment, and measure 45 on an average. They occupy the central portion of the proglottides (fig. 2). Vas deferens. The vasa efferentia appear to unite in a small globular structure, evidently a vesicula seminalis, which lies in the anterior of each proglottis, near the middle of the segments and just at the mesial end of the cirrus pouch. They are in no instance conspicuous, and do not appear to function for storing the spermatozoa, except in the earliest stages; as the cirrus develops, the vesicula atrophies and finally disappears. That portion of the vas deferens within the cirrus pouch becomes increasingly coiled as the seminal vesicle atrophies, and it appears to take on the functions of the latter. Cirrus pouch. The cirrus pouch is a relatively large, thick- walled sac, lying diagonally across the antero-lateral angle of the segments on the pore side, and it opens with absolutely regular alternation on the lateral borders of the segments, slightly in front of the middle. It measures about 450 long and 150 broad, and as the worm is at this time only about 1 mm. broad, it is a conspicuous organ. It persists in older worms after the testes are gone and the female glands are well developed; but at this stage 6s the worms are about 2 mm. broad, and as the cirrus pouch does not increase above the dimensions given previously, its size in relation to that of the proglottides is not so great. The cirrus is nearly always extruded; it measures about 400” long and gop thick at the base. In the early stages it is a long, fairly thick tubular structure, slowly tapering from base to bluntly rounded tip. Its external surface is thickly covered with backward- curving spines about 4m long (fig. 2). It runs posteriorly as a rule, and in many cases is recurved, so that the tip points towards the lateral border of the same or the succeeding segment. In worms in which the female genitalia are developed, the cirrus is generally stouter and more conical in shape, and it tapers much more rapidly (fig. 3). Ovary. The ovaries are only found in older worms. They first appear about the seventy-fifth segment, and have reached full development by about the ninety-fifth segment. In these worms the seventy odd small segments in front of the one in which the ovary is first seen are devoid of all traces of genitalia, either male or female. Apparently therefore, in attaining a certain age, the worms lose their power of developing reproductive organs, any further segments being sterile. Fully developed ovaries are about 700“ broad and consist of two lobes each composed of lobules running for the most part laterally. The two lobes are united across the mid-line by an isthmus, they are unequal in size, the one on the cirrus side being only about half the size of the one on the opposite side. The result is that they present a regularly alternating asymmetry (fig. 3). In addition, there are three or four small masses of ovarian tissue lying anterior to the isthmus of the gland, and more or less directly connected with it. The ovaries lie across the centre of the proglottides about mid-way between the anterior and posterior borders. Recepltaculum and vagina. Both these structures are absent. Vitelline glands. The vitelline glands consist of a small horseshoe-shaped mass of tissue, with the concavity facing forwards. They lie in the mid-line behind the ovaries and towards the posterior margin of the segments; ducts can be seen running forwards from their concavities, and these have the follicles of small shell glands grouped around them. 66 Uterus. The uterus first appears as an oval tubular ring nearly completely surrounding the ovary and vitellarium, which rapidly atrophy (fig. 5). Outpocketings soon appear on the tubular uterus (ig. 6). They become progressively larger and more complicated as development proceeds, until the ring-like structure is nearly lost, and in full development the uterus is represented by a large lobulated sac occupying almost the whole of the proglottides, both laterally and antero-posteriorly (fig. 7). Fic. 5. G. australicnsis. Segments showing first stage of uterus. ov., ovary undergoing atrophy; wf., uterus. 35. If A.M_B.del Vig. 6. G. australicnsis. Segment showing uterus more fully developed. p., uterine pore. X 35- Another remarkable development is that of uterine pores; these open externally on the centres of the posterior borders of the segments, one on the dorsal and the other on the ventral surface. From examination of horizontal sections, we are of the opinion that these pores arise from a single central opening on the posterior of the uterus, from which two canals run, one to each pore (figs. 6 and 7). 67 Eggs. The eggs are slightly oval with blunt extremities. They measure about 65 long and 52, broad, and the contained embryo, which is enveloped in an albuminous covering, is also oval, and measures about 36” by 26”. The hooks on the embryo are about 16m long. Fig.7 A.M.B del. Tic. 7. G. australiensis. Yully developed uterus. ., uterine pore. x 35. DIAGNOSIS. Although the hooks had been lost from our specimens there seems no reason to doubt that the worm is Gyrocoelia australiensis, Johnston. It is, however, necessary to point out that Johnston (1912) figures five minute testes lying immediately anterior to the ovary. Our specimens present a similar appearance, but these structures seem to us to be detached ovarian acini. Further, Johnston (1914) recorded a Gyrocoelia sp. from L. lobatus, which worm he obtained from the Australian Tropical Institute, the same source as our material, so it is practically certain his unnamed species is also G. australiensis. Clausen (1915), in his description of G. paradoxa (von Linstow) (= Brochocephalus paradoxus), figures a bi-lobed or double receptaculum seminis enclosed in the uterine ring. Our specimens present a somewhat similar appearance, but the structure is obviously the degenerate ovary and vitellarium. The occurrence, in our worm, of the male and female sexual organs at different times, which results in a strobila being male when young and female when middle aged, raises the point as to whether 68 this condition does not likewise exist in the genus Déoicocestus, Fiihrm., 1900, in which case its characters would be limited to the possession of double male genitalia, and an irregularly alternating vagina. REFERENCES Crausen, E. (1915). Recherches Anatom. et Histolog. sur quelques Cestodes d’O’iseaux. These. pp. 63-75. Neuchatel. Jounston, T. Harvey (1912). On a re-examination of Krefft’s species of Cestoda. Records of the Aust. Museum, Vol. LX, No. 1, pp. 28-32. —— (1914). Second report on the Cestoda and Acanthocephala collected in Queensland. Annals Trop. Med. & Parasitol., Vol. VIII, No. 1, p. 108. 69 A CONTRIBUTION TO THE KNOWLEDGE OF THE BIONOMICS OF SAND-FLIES BY Capt. JAMES WATERSTON, R.A.M.C. (T.) (Received for publication 17 February, 1922) PLATE VIII During the late campaign in Macedonia, members of the B.E.F., Salonika, suffered more or less continuously during the hot season from sand-fly bites and sand-fly fever. As seems to be always the case, the trouble was very definitely localised, the usual foci being either dilapidated villages in or near which units were stationed, or entrenched positions which had been long occupied. In the summer of 1918 there was a severe outbreak of the fever in and round Janes (where in one unit from mid-June to mid-August there was an incidence of 150 per cent. of the total strength), and I was instructed in the third week of June to investigate the conditions as regards sand-fly in the district. Owing to an attack of dysentery I was unable to proceed to Janes until the second week in August, when much valuable time had unavoidably been lost. The following pages reproduce almost verbatim a report on my investigations between August 12th and September 26th based on copious notes made on the spot. There have been incorporated also various observations made during 1917-1919 on PAlebotomus in Macedonia. It may not be irrelevant to mention in passing that on my way to Salonika in July, 1917, I kept a constant lookout for Phlebotomus wherever the frequent stoppages of a troop train offered an opportunity of searching any likely spot. A species of the genus (probably fapatasi?) was first encountered at a small station near Beaune (Cote d’Or), and other examples were noted later at Orange (Vaucluse). At the time I was not prepared to find the genus so far north, but Major Joyeux has subsequently told me that during the war special search was made for Phlebotomus in France, with the result that P. papatasii was met with amongst other places near Beaune and as far north as Paris itself. In Macedonia the species of sand-fly (Phlebotomus) investigated 7O were three in number, viz., P. papatasii, Scop.; P. minutus, Rnd., and P. perniciosus, Newst., the first named occurring, at Janes, in by far the greatest numbers. Owing to the difficulty of discriminating between the females of these insects no attempt was made to determine specifically all the material collected, but in some hundreds of cases in which identification was effected P. minutus formed about 1 per cent. and P. perniciosus 2 to 3 per cent. of the total. Probably one is safe in saying that 95 per cent. of the sand-flies observed belonged to P. papatasiz. Melanic forms of all three species were observed and in long series of papatasii many colour variations were noted, but the proportion of these to the total was not worked out. Really dark examples were, however, uncommon. From various sources it appears that by the date on which observations were commenced sand-flies were much less common than earlier in the season (June). Nevertheless, up to September 1oth they were numerous wherever sought for, and in certain localities very abundant. Between the 1oth and the 14th of the month there was a great falling off in the numbers found daily, but even so a good many could be taken up to the end of the month, and these later captures were still persistent in their attacks. In connection with the association together and _ seasonal prevalence of these sand-flies, it is interesting to note that in 1917 (August to September) I did not certainly meet with P. minutus, while P. papatasizi and P. perniciosus (though one or other occurred in numbers at Kalamaria, Salonika, Karasouli, Lahana, Nigoslav) were never taken together. In the present season, 1918 up to June 24th, only perniciosus occurred sparingly at Kalamaria. Again between July 24th and August toth perniciosus (with one or two minutus latterly) was so abundant in the same place that in a single evening (9 to 11 p.m.) two hundred could be taken at light in one latrine. P. papatasii, though carefully sought for, was not detected. Newstead, in describing perniciosus, records a similar experience in Malta. ‘Two examples of P. minutus were found in association with this species, but strange as it may seem, not a single example of P. papatasit was either captured or seen on these occasions.’ In 1917 sand-flies (P. perniciosus) were still common at Karasouli in the third week of October. 71 As to the part played by these three species respectively in carrying sand-fly fever in Macedonia, little evidence is to hand. Assuming that all are potential vectors, it is by no means certain that each carries the fever in proportion to its numbers. In any case P. minutus seems hardly likely to be of much importance in this réle. As regards papatasii and perniciosus, the former is in my experience not only more active but more voracious and incessant in its attacks. In three localities where sand-fly fever was reported (Karasouli, 1917; Salonika, Janes, 1918) it was present in numbers, and in two instances was the only sand-fly taken. My present impression is that ferniciosus is if anything a less efficient vector than fafatasii, but the point is one which can be settled only by direct experiment. The work done at Janes has included the following :—(a) A search for the early stages of sand-flies im situ. (6) A study of the habitats and habits of the imago. (c) Breeding and rearing of the flies. (d) Preventive measures. A. SEARCH FOR THE EARLY STAGES During the whole of my stay at Janes search has been made more or less continuously for ova, larvae, or pupae of sand-flies. In the first three weeks practically nothing else was attempted, but the result of protracted examination has been negative. Search has been made (1) in the soil itself to a depth of 6 inches, particularly near any indication of moisture; (2) along the sides of earth cracks and fissures so far as they could be followed; (3) beneath loose stones and in the superficial layer of cracked rock at or near out- crops; (4) between sandbags and loose earthy rubbish thrown up in erecting tents, marquees, etc. ; (5) between mud bricks or masonry in the lower tiers of buildings; (6) in the dampest looking recesses of dug-outs and among débris beneath hay stacks. This failure to find any early stages is parallel to the experience of many other investigators, and does not necessarily imply that search was made in wrong directions. Subsequent experience of ova and larvae from captive females indicated that the earth, etc., previously examined was much too dry, and that larvae, if they were to be found, should have occurred at greater depths. It is 72 possible, however, that larvae are absent or much less plentiful while the adults are numerous. Newstead, who worked in Malta in July, August and the first week of September was under the impression that larvae would be more numerous in autumn about a week after the adult had disappeared. As has already been stated, sand-flies began to decline in numbers at Janes between the roth and 14th of September, and it was on the 20th that I found newly-hatched larvae for the first time in my breeding-boxes (see below, App. III, No. 1). B. HABITATS AND HABITS OF FLIES Habitats. (1) In mid-August, and for some time afterwards, sand-flies occurred practically everywhere in and near the C.C.S. They were more numerous, however, in tents and wards. In dug- outs they were comparatively scarce, which was at first rather surprising since at the same time one had a report that up the line the flies swarmed in dug-outs. The explanation appeared to be simply that the flies gathered as near as possible to their hosts. The hospital dug-outs were unoccupied, while the others were regularly used. In the same way where two wards, one occupied the other empty, adjoined, the former yielded many more flies than the latter. Again in a ward which, as sometimes happened, had only a corner bed occupied, the flies were most numerous in that corner. In large marquees or wards the distribution of the flies varied from day to day and with the hour of the day. After a windy night few could be found, and those that occurred were in sheltered corners. They were most abundant after still, damp nights. In the morning one found them mainly beneath the flaps running round the top of the sides of the tent, and the flies so taken showed a large percentage of newly-fed, gorged females. Later in the day, from roughly 11 or 12 a.m. to about 2 p.m., the flies became temporarily scarcer, while during the afternoon they were more frequently seen in numbers on the lower half of the side, males being numerous and often predominating. Pairs 7z cop. were common in the morning. Where the sides of marquees, closed during the night, were left standing by day more flies were found. Invariably, too, where the sides were rolled up into a corner, flies 73 were found within the roll. When the ground at the corner was cracked, flies were observed emerging from the earth and entering the folds of the roll, so that even after clearing such a roll in the forenoon one might later in the day take many flies from the same place. (2) Earth cracks, in fact, proved to be effective day-shelters for Phlebotomus. The most important proved to be those occurring on the exposed surface of the soil where it had been dug out to make a level floor for tentage. The presence of flies in the cracks was easily demonstrated by blowing tobacco smoke or by squirting into the lower portion a little paraffin or even plain water. Within a tent twelve to fifteen flies might thus be driven from a crack less than a foot long and a few inches deep (August); when so expelled the flies generally came back after a short flight, and they had to be pretty thoroughly disturbed before they would travel any distance. Sometimes when driven from one crack they merely hopped along the cut surface and entered another. (3) Sandbags employed to raise the level of the sides of tents, etc., are also a fertile source of trouble. The flies rest not only in the crevices between the sacks, but also enter the loose earth inside through the interstices of the coarse sacking. Where dug-outs are near wards, and their entrance is reinforced with sand bags, many flies will be found. In the tent which I occupied during my stay at Janes were several courses of sandbags. The following experi- ment was many times verified. When one opened up the tent and allowed strong light to fall so that one side of the person was illuminated and the other side in the shade, as one stood with the hand extended towards the sandbags, then though no flies had previously been noticed, bites began to be received in a very short time. After one or two such trials it was possible to see the sand- flies emerging from their retreat to the attack, which was always made on the shaded side. Besides the situation indicated, sand- flies shelter by day in hanging clothes, cupboards, blankets, beneath pillows, etc. Habits. In studying the habits of Phlebotomus, much difficulty was at first experienced in keeping the flies alive. In test-tubes many died within two days, and few survived the third day. Not even when the air was kept humid could oviposition be induced. 74 In the end, suitable apparatus for handling the flies was improvised, and a list is appended. Various devices for keeping the flies alive were tried during the first four weeks, and ultimately earthenware pots for single or smaller lots and cages for larger numbers proved most useful (see Appendix I). In the earthen pots—which were first suggested to me by Lieut.-Col. C. M. Wenyon—single specimens were easily kept and handled as follows. The fly was caught in a test-tube over the end of which a piece of cotton was loosely tied so as to form a small bag. The test-tube was then inserted through the sleeve of the cover. A little water was next poured into the pot and the cover tied on, the test-tube being fixed at any desired height above the water by a piece of string passed round the sleeve (fig. 4). Flies so kept showed an interesting periodic movement, staying all day in the cool, humid air below and appearing at the top of the tube during darkness. They were generally restless—particularly papatasii— about dusk. In cages provided with a tray filled with moist earth, stones, etc., a similar movement was observable, the flies hiding in crannies by day and emerging by night. A considerable proportion, however, remained by day crowded together in corners of the cage. They were frequently observed drinking from the wet earth. In collecting sand-flies a pot was taken and covered and the flies introduced into the sleeve from the test-tube in which they had been caught. The sleeve was then tied and the pot stood in a bowl of water (fig. 5). To feed them it was necessary, first, to turn the flies loose for a time into a fly-proof box fitted with a sleeve to admit the arm (fig. 3). It was noticeable that flies kept in solitary confine- ment would seldom feed if the tubes were merely inverted over the arm. In pots, unfed, a variable proportion of the flies—up to 50 per cent.—lived a week, 2 to 3 per cent. lived nine days, one @ (out of a batch of six hundred) lived exceptionally thirteen days. No © was observed to live more than eight days under these conditions. All the females that lived nine days or over were examined as to the condition of their ovaries, and were found to be spent or practically so. In no instance could they be induced to feed in this state, though offered repeated opportunities—in the case of the last extending over the eleventh, twelfth and thirteenth days. 75 (1) Biting, etc. No males were detected in this act. The large number of this sex found with the females (as compared with Culicidae, where often the total catch from a ward will not include a single ¢ ) seems to be due to the fact that mating takes place after the female has had a blood feed. In all the couples examined the female had recently gorged herself. While feeding Phlebotomus is easily disturbed, the slightest movement of the skin being sufhcient to put the fly to flight. It is thus difficult to study the process of biting in detail. The insect settles, pitches forward slightly, thrusting the somewhat stout rostrum downwards while the palpi (maxillary) diverge a little. I have not seen more than about one-third of the rostrum enter the skin, and the labium does not buckle up as in Anopheles when engaged in the same act. Only the labella are flattened out above and behind the piercing part of the associated organs. The wings are meanwhile poised ready for instant flight. Blood can be seen in the sucking stomach within sixty seconds. A full feed on an empty stomach occupies from four to four and a half minutes, at the end of which the fly suddenly withdraws the rostrum and makes off. For about forty-eight hours after a meal blood can be seen in the ‘sucking stomach’ over-lying the mid-gut (whose contents are brown or blackish) on the left side, and the whole gut may be cleared in five days, but the process generally takes longer. The females will feed at two-day intervals (possibly at shorter periods), taking less copious feeds, but attempts to find how many feeds could intervene before the completion of oviposition were abandoned. The digested blood is passed out ultimately in the form of small, dark, sticky drops; one or two specimens when captured had the stomach gorged with pale fluid and the gut hardly darker. (2) Movements. In cracks and on rough earth Phlebotomus ordinarily proceeds by short runs varied by jumping to one side or another; on smooth walls or fabric more by jumping and _ short flights. Confined in a test-tube the males commonly mount in seeking an outlet. Females, if recently fed, on the other hand settle downwards, but after the gut has cleared and the eggs laid they behave much as do the males. When first introduced into a cage or confined space sand-flies make determined efforts to escape. They pass through astonishingly small openings, using the proboscis 76 apparently as a lever and emerging sometimes with a considerable proportion of the scales and hairs missing. (3) Drinking. As Phlebotomus runs over the surface of a piece of earth it may sometimes be observed to plunge the rostrum into the earth. If there is moisture present the fly may remain for some time still. To observe what was happening, specimens were isolated in plaster of paris cells and watched under a Zeiss binocular. After they had been imprisoned for twelve hours the block was placed in clear water, which at once mounted to the floor and sides of the chamber containing the flies. Practically all the insects commenced to drink, some of them running about excitedly for an instant before settling. As was noted before, the tip of the rostrum (labrum, epipharynx and hypopharynx, etc.) was thrust distinctly into the plaster, and the labellac, flattened out, closely appressed to the porous surface. The short superiorly ensheathing maxillae generally moved backwards and forwards for a short time in front of the clypeus, and the maxillary palpi—rather widely divergent, nearly at right angles to one another—moved tremulously. But during drinking the mouth parts were still. Swelling of the abdomen could be traced as the drink proceeded. Flies, both sexes, were found to drink readily in this way about twice daily. They were tried with drops of fresh human blood on paper slipped into the cell, but without result. This can hardly be regarded as conclusive, as the blood dried rapidly. Defribrinated diluted blood was next allowed to soak into the cell containing flies, but did not prove specially attractive. The flies appeared to be able to extract only diluted serum—the corpuscular débris being retained by the plaster. In the same plaster cells cut deeper Axopheles was also induced to drink water. C. BREEDING AND REARING OF FLIES (1) Oviposition. While the conditions in pots standing in water were congenial to the adult flies, there was apparently insufficient moisture within the pot to induce egg laying. Nor did females kept in a tube over moist paper oviposit. Complete success was, however, attained by the following methods. A thick ‘tray’ was partly filled with fragments. of earth over which was lightly 77 sprinkled some crushed faeces of lizard, rabbit or man. The tray was then soaked in water and placed in a pot into which water had been poured to a depth of a quarter of an inch (fig. 5). The flies were then put in through the sleeve as usual, and the pot stood in water. After an interval the pot was opened; on washing out with as small a quantity of water as possible (20 c.c. was sufficient as a rule) a number of eggs were got from the sides. They were collected as follows. The liquid was centrifuged and the clear part used to rinse the pot again. This was done three times. Finally the sediment containing the ova was pipetted off and the ova themselves separated out under the microscope. The percentage of fertile eggs was extremely high, and their viability was not appreciably affected by their being for some time in water. Besides the ova taken from the sides of the pot many were to be noted on the tray or among the earth, etc. To recover eggs so laid was an extremely tedious process. Some were picked up with a brush by direct observation, others recovered after carefully washing the contents of the trays. An improvement in gathering ova from the sides of the pot was effected by lining the inside with a single piece of cotton pressed down to fit exactly. The pot was then loaded as before. The eggs (which are at first pale and then darker like those of mosquitoes) showed up well against the cotton which was cut into strips and so passed below the microscope. A further advantage was that the cloth showed definitely the average position of the eggs laid on the sides of the pot to be along a band about three- quarters of an inch above the water surface (fig. 5, 0.) This seems to indicate that there is a fairly precise optimum as regards moisture for oviposition. Eggs on the sides of the pot occurred singly, but the majority were found below the earth close to one another in bunches. The females had crawled into the smallest crevice possible and appeared to have projected the eggs still further. Beside all the larger collections of eggs in these cracks the mother could be found within two millimetres. She lay, as a rule, flattened between the surface of the tray and the overlying earth, with sometimes one or two eggs between the extremity of the abdomen and the rest of her laying. Sometimes an egg was seen attached to the terminal bristles of the genital appendages, and once or twice females had died with the egg between these appendages or blocking the genital 78 atrium. With a little care, it was easy to clear the earth from round each dead female and count the eggs laid. The larger batches contained from twenty-seven to thirty-four eggs. Sand-flies are not prolific insects, and the total ovarian content in ?. fapatasit runs from forty to fifty eggs. Assuming that these females had previously deposited one or two isolated ova on the sides of the pot, the proportion of eggs actually laid to the total possible is a high one. In some hundreds so secured there were practically none that failed to hatch. This high fertility and the large number of eggs laid may reasonably be held to indicate that natural conditions for oviposition had been secured. Newstead, who in 1910 watched egg laying of P. papatasz in Malta, states (2) that the egg is projected some distance from the abdomen of the female; (@) that the process is so exhausting that the female may die after it; (¢) that most of the eggs were laid below the moist blotting paper supplied to induce oviposition. With these notes the foregoing observations are quite in harmony, and it seems probable that the violent ejection of the ova serves to insert them into crannies where the abdomen of the mother cannot penetrate. Newstead saw the ova thrown about three times the length of the female abdomen, and this again agrees closely with the distance noted by myself. In cages where the earth, etc., was placed on a shallow tray no excessive moisture ran up the sides (the water being completely absorbed), with the result that eggs occurred, so far as could be seen, only in pockets with dead females close by. An attempt to determine whether the sand-flies in ovipositing showed any preference for one kind of faeces was inconclusive, owing to the development of microfungi, whose ramified mycelia enmeshed the eggs so that a complete count was impossible. The fungus growth was slight on lizard faeces either by themselves or mixed with earth, but very abundant on human faeces. So far as one could judge, however, plain earth or earth and lizard faeces held more eggs after some hundreds of females had been allowed to oviposit on plaster trays giving a choice of several kinds of larval food. No female was noted to oviposit till four or five days after capture, and the eggs did not hatch till at least nine days more. The incubation period probably varies seasonally and specifically. 79 Ova belonging to at least two species were secured. Of these, one, the larger, was by its size, abundance, texture and resulting larva plainly P. papatasii. The other (probably P. perniczosus) was shorter and a little stouter. Apart from size, the two could easily be told towards the end of the period of incubation by two dark parallel lines which appeared shining through the shell laterally in each. In papazasii the lines at the caudal end of the ovum were rather broader and tapered towards the head. They could also be traced round the head and back towards the tail for a short distance. In the second species the calibre of the lines altered little between tail and head, and they extended nearly half- way back towards the tail on the other side. After hatching, these lines could be recognised as the caudal bristles, which are relatively short in papatasii, All through the first instar there is a kink in the caudal bristles, indefinite and near the extremity in papatasii, and much more decided and further back in the second species. The kink is, of course, at the point where the bristle is bent within the egg. For the health and ultimate hatching of the egg a considerable degree of moisture is necessary—rather less, however, than is required to induce oviposition. Excess of moisture for a limited time has no effect on the viability of the eggs, for they may remain immersed a day or over and yet hatch. Eggs resting on earth surrounded by a thin film of water for three to four days also hatched. Drying, on the other hand, even a short time is fatal. A batch of eggs exposed in the shade overnight shrivelled before I1 a.m. next day, while in the daytime a few hours brought about the same result. (Time, 20th-25th September.) (2) Larvae. When the egg is about to hatch the caudal bristles may be seen to move. There is a rippling movement also of the body from the tail towards the head on which, high up, almost vertical in position, is the well-developed dark egg tooth. Dehiscence of the shell is affected by a cut extending sometimes to half the length ventrally (?) and backwards for a short distance dorsally (?). The eyeless and legless maggot emerges slowly, and is at first entirely pale save for the egg tooth and bristles. The head, however, darkens in a few hours. In emerging the larva seems to be coming from two valves, but in some cases the line of 80 dehiscence is cut laterally as well as dorso-ventrally so that a relatively large oval piece of chitin falls from one side of the shell. Excessive moisture retards the process of hatching, always a slow one, and when the surface on which the egg rests has a thin covering film of water the larva may be found barely clear of the shell twenty-four hours after hatching began. The newly hatched larva is sluggish, and, indeed, during the whole of this instar little activity is shown. It lies either flat on the supporting surface with the caudal bristles extended in the same line, or resting on the ventral caudal third to one-half of the abdomen, with the rest of the body raised and the last segment with its bristles slightly upturned. In this pose the larva is U-shaped. In a modification of this attitude the head is again thrown forward and the whole creature in profile S-shaped. The larva’s progress is undulating. The head is first raised and the anterior segments stretched forward. The mandibles now press (or grip ?) firmly some inequality of the surface, and with this purchase the body is dragged slowly forward. Folds in the skin, and possibly the peculiar hairs of the body, aid in the process. The tiny larvae begin to feed almost as soon as the head has darkened, 7.e., the mandibles are hardened. In feeding on lizard faeces they select the rough portion consisting of chitinous fragments mixed with partially digested fibre, and reject the more homogeneous limy part. Three individuals watched settled down respectively to the mandible of an ant, the head capsule of some small hymenopteron and the leg of a beetle, and ate the half- digested muscle fibre inside these structures. In the same way they enter and feed on the dead bodies of the parent flies. Older specimens of the first instar will also attack and devour larvae immediately after hatching. Fungus was so trcublesome (being rightly or wrongly blamed for the loss of many young larvae) that some substitute food was sought. Finely ground mixed blood and earth has proved satis- factory for this purpose. It is spread in lines on shallow plaster trays with the eggs near, and the whole covered with rough pieces of earth. The young larvae feed readily and appear to thrive, and no fungus has as yet developed. Larvae in the first instar can survive excess of moisture for some 81 time. They sink readily in water, and are not sustained for any length of time by the surface film. When dried after lying twenty- four hours on wet soil they are lethargic but soon recover, yet like ova they are extremely sensitive to thorough drying and shrivel if exposed a few hours in the shade. The first ovum to hatch did so on 20th September, and the first example in the second instar was seen on 26th September. In this stage the egg tooth has, of course, gone; the caudal bristles have increased from two to four, with a dark chitinous saddle connecting them. There is also a considerable increase in size. In India, according to Howlett, there are separate broods of P. papatasii in August and September, while the wintering brood begins in late October or early November. At Janes I have seen no trace of the September brood, and the slow growth of these larvae hatched from late September eggs suggests that in Macedonia there may be one less brood than in India, and that the wintering brood commences here at least a month earlier. Subsequent experience confirmed my expectation that the September hatched brood would hibernate. It was impossible to give much time to observing the larvae during the last week of September and the first fortnight of October, owing first to the congestion of the C.C.S. after the push, and later to the severe epidemic of influenza. But about the middle of October a consider- able number of larvae, mostly in the second instar, were successfully taken to the Base and installed in the laboratory at 52 General Hospital. They were by this time very sluggish, and, in spite of being kept in moistened earth in a room which was heated at least during the daytime, latterly ceased to feed, while some died. One lot which had been left as it came from Janes was allowed inadvertently to dry up completely. On examining this tray in the third week of November (by which time it must have been quite dry for a month) I could find no larvae moving on the surface, but on breaking up the earth as a precautionary measure before throwing out the contents of the tray, I found several larvae in little pockets or blisters in the earth. They lay quite straight out but appeared stout and contracted, the integument wrinkled slightly towards head and tail, the gut empty or practically so. The only movement exhibited was a slight twitching from side to side of the extremities 82 when the animal was gently touched. By bringing larvae in this condition into moist, warm surroundings it was possible to revive them so that they recommenced to feed. Unfortunately the few experimented with were accidently destroyed. The bulk of my larvae had not revived naturally at the end of March, 1919, and the attempt to bring them to London failed. While it is not suggested here that Phlebotomus hibernates naturally under such conditions as have just been described (the opposite, indeed, is much more likely), it is evident that these larvae, once the critical stages of hatching and the first instar have been over- come, have unexpected powers of resisting dessication which must be of considerable help to the species in its natural breeding haunts in cracks, etc., where the conditions as regards moisture are variable. D. PREVENTIVE MEASURES (1) Nets. During my stay I used the sand-fly net issued to the troops with satisfactory results. With a flash-light at night one could see the flies settling on the net, yet none were noted to pass through. Once or twice sand flies (up to four) were found within the net in the morning, but these, I believe, came not from the outside but from blankets or below pillows, having got there during the process of bed-making. They also find harbour inside sleeping bags, and all bedding should be thoroughly shaken or beaten just before the net is adjusted for the night. In this connection, however, it is worth recording that of thirty to forty mixed sand-flies placed in a small bag of the material from which the nets are made and hung up loosely, all escaped within half an hour. The flies behaved as they do when confined in a cage, pushing in all directions to find an outlet. I have found in the same way that a certain number of mosquitoes in a brood covered over with ordinary netting will manage to struggle through. The species in which this was noted were Culex pipiens, Anopheles palestinensis and A. bifurcatus. In spite of this the net in use appears to be efficient, though possibly where P. minutus pre- ponderates a finer material might be necessary. 83 (2) Repellents. (a) Ordinary Paraffin is, if liberally applied, effective in keeping off the flies. It might be used as a stop-gap. M.T.C. drivers reported that they had employed it with good results, but I do not regard its regular use as advisable, and made no tests personally of the time for which an application remains effective. (6) Mosquito Pomade and (c) Paraguit were both tried. The former was issued to various members of the personnel of the C.C.S., who reported favourably on its use. Of the two, Paraquit is perhaps more pleasant to use. Well rubbed in, I found it effective for about three hours when one was sitting still or not moving actively. I found it sufficient to rub the Paraquit into the back of the hand, wrist and halfway up the arm towards the elbow, the neck, ears and round the scalp, but not into the hair. For the bites themselves I found appreciable relief by dabbing on a little rectified spirit with cotton wool. (3) General. The breeding-sites of Phlebotomus hitherto recorded have been varied. Larvae have been found in soil along the sides of the embedded portion of the lower corners of old masonry; in rubbish in cellars, in earth cracks, etc.—the common feature in each case being darkness, moisture, and the presence of organic débris. While it is possible that at 31 C.C.S. the flies came in part from farm buildings near by, I think it most probable that the main breeding-places were in or round the hospital itself, in the cracks where the adults were themselves found. There was nothing in the distribution of these adults to suggest that their breeding-sites were other than generally distributed. Where there were more flies there were more men sleeping. In such circum- stances prophylactic measures are difficult to carry out, and the only remedies available are probably palliative rather than radical. The following suggestions are given for the treatment of tents and marquees. The floor should be levelled and cracks filled up with a mixture of cvesol and sand or sawdust. Clay is unsuitable, as it is apt to crack in turn (this was repeatedly noted) and the most minute fissure will harbour many flies. The floor should then be liberally watered with a strong solution of cresol, and if possible covered with a ground sheet. Periodically, according to the severity of the plague, the tent should be closed and sprayed with a solution of formalin or fumigated with cresol. 1 per cent. 84 formalin has been recommended for spraying, but I believe this is too weak. In corrugated iron huts a 1 per cent. mixture of cresol in paraffin emulsion might usefully replace formalin for spraying. Around the tent all cracks (to a breadth of two feet from the tent) to be filled up and the soil to be sprinkled to the same width with cresol. In latrines where crude cresol was experimentally sprinkled ° round the drums, sand-flies were reduced in number for two to three days, and ceased to bite on the exposed legs though bites were still received on the neck. A similar reduction was noted in a store where cresol was regularly sprinkled. As far as possible, shallow soil with frequent outcrops of loose friable rock should be avoided in choosing a camp site. Loose soil removed in pitching tents should not be allowed to remain in camp. Building up with sandbags round tents, etc., is to be deprecated. Spilling of water or any liquid containing organic matter should be carefully avoided. The appearance of cracks in the soil should be watched for and counteracted by filling up when possible and spraying with cresol when the cracks are more extensive. From ten to fourteen days after the disappearance of the successive waves of sand-flies that occur from June to September a thorough sprinkling of cresol over suspected breeding-sites is to be recommended. I have to thank heartily Lieut.-Col. Ievers, D.S.O., O.C., the 31st C.C.S., for his kindness in affording facilities for work during my stay at Janes, and very specially Lieut.-Col. C. M. Wenyon, C.M.G., who first suggested the investigation. In conclusion, I should like to record my indebtedness to Captain Beer and Corporal Gibson, R.E. (143 Field Company) for carrying out my design for the breeding-cage (fig. 1), and to Captain Morrell, Dental Officer to the C.C.S., who supplied the plaster trays and cells. 85 APPENDIX I AppaRATus UsED IN StupyING THE Lire History or Phlebotomus spp. Caczs may be made to suit requirements. (a) The following dimensions housed comfortably 600 sand-flies, 13 in. broad 14 in. high x 6 in. deep, made of 1 in. wood, back of tin driven into the wood and nailed, on inside faced with cotton glued down and whitewashed. All joints glued and rabbeted. One or two panels of cotton fixed closely by strips of wood to afford ventilation. Tin tray to hold water in bottom. Flies admitted by tightly-corked circular opening at side. Front glazed to depth of about ro in.—the glass set in putty. Narrow panel door below glass 12 in. x 2 in., made fly-proof by cloth edging. Allround the door the joint is rabbeted and felt lined. The door may be Fic. 1. Large breeding and feeding cage (11} in. X 15}in. X 14}in.) front view. Glazed in front. b—Rabbet-fitting door giving access to tray for disposition of plaster receptacles of earth, etc., on which flies oviposit. The mid-partition cut down to give access of flies to host. a—Detail of ventilation showing the inner screen of stout wire-gauze and the outer cotton cloth. secured byaclip. The middle surface of the rabbet pressing against the felt makes the joint fly-proof. (4) The above cage (no fig. given) is suitable when ova and larvae only are wanted. When the adult flies have to be fed for some time in numbers a more elaborate cage is required (figs. 1 and 2). It consists of two chambers. In the right-hand one the flies oviposit on material in tray. In the left chamber a rabbit or other small mammal can be accommodated for a time, but should not remain continuously with the flies. The animal sits on a wire grating and its urine and faeces are caught in a tray. Ventilation at top and side. In the latter 86 case the ventilator is double—cotton outside and strong wire-gauze towards the rabbit. Inside the door and fitted to the sides is a large sleeve through which the animal is introduced to the cage. In the middle the sleeve is held closed by an elastic belt to prevent the egress of the flies. The rabbit is introduced by forcing its head through the confined middle of the sleeve and sliding back the band over its body. The flies are put in as in cage (a) and gain access to their victim over and below the mid-partition, which is cut back for this purpose. Fic. 2. The same from behind, showing the grid on which the rabbit is accommodated. Below, next to the door, is a wooden bar with a felt extension to make the faeces and urine tray fly-proof. The space above the grid is sleeved. Frepinc Cacr. P= = wa - ie 4 6 2 SMALL CYSTS OF &£. HISTOLYTICA Small cysts (below ton) of E. histolytica have been noted by James (1914), Woodcock and Penfold (1916), Wenyon and O’Connor (1917), Dobell and Jepps (1917), and others. A detailed account of them, and evidence of their differentiation from the larger cysts of E. histolytica, are given by Smith (1918 and 19109). Most observers hold that these small cysts constitute a separate “strain ’ of E. histolytica, but Woodcock and Penfold state that it is quite likely that this form is either a distinct species or distinct variety. Morphologically they are similar to the ‘ordinary strain’ of E. histolytica, except only in size, and it has been generally assumed that they belong to the same species. No work appears to have been published on their pathogenicity to animals. Dr. R. M. Gordon and I endeavoured to infect kittens with this small cyst without success, but as failure to infect controls with the large cyst also occurred, no conclusions could be drawn. As some doubt, therefore, exists regarding this so-called ‘ small strain’ the findings of the two sizes have been recorded separately in Table I. In Table IV these figures are combined for comparison with the findings of other observers. Little difficulty was experienced in ascribing infections to their respective groups as very few cysts in the neighbourhood of Io were encountered. In eight cases cysts belonging to both‘ strains ’ were present. 96 Table III has been compiled from a paper by Smith (1919) with the addition of the present series and shows the relative proportions of ‘ small ’ and ‘ordinary strains’ among the total E. histolytica infections. The figures for the two Mandos groups—troops and children—have been added together as the distribution of the two sizes is similar in each group. Attention is drawn by Smith to the small percentage of the ‘ small strain ’ in persons who had not been out of England. Taste III. Size of Cysts in E. histolvtica Infections. England only England and Abroad _ | Amazonas Matthews | | Dobell | Matthews and Mackinnon |M ackinnon and and Present Smith (1918) Jepps Smith Series (1917) | Dysenteric | Non- Chronic | and non- | Dysenteric Dysenteric, Troops dysenteric | dysenteric | dysenteric | _ cases cases and cases cases cases | Children | | Cases 98 56 | 209 200 306 125 Infections ... 99 59 225 215 325 133 ; “ Ordinary’ % 85 64 47 65 66 60 “Small” % 15 36 53 35 34 40 Table IV shows the findings of intestinal protozoa by various workers in different parts of the world. The figures represent percentages, and are all based on the results of one examination per case. The figures for E. histolytica include all ‘ strains’ above and below roy, excepting those for Queensland where, Dr. Maplestone informs me, no cysts below rou were found. In the latter instance the stools were three to fourteen days old when examined. Professor Kofoid has kindly supplied me with the figures for the United States of America. He states that they are probably higher than normal in the population as they contain large numbers of foreign immi- grants and negroes from Florida. Figures published by Kofoid, Kornhauser and Plate (1919) for overseas troops of the United States Army are somewhat higher than those for home service troops. 97 *sasvo PHI uy |. ssasvo tZ Uy y z.gh 0.0 +. sdoory, satag quasar spuozRuly 41.98 L.0€ 0.0 Z.0 48.2 ZT o.F1 +.9 Ale £.6z oe 6.98 I.br S.zz 6.£ 6bz 9£S aay sdoory, yooyss ADTAIOS jo awoH worPTYO PIOjOM ‘v's'n + -- + aoe aoe wee ee susho01sv] gg aoe fe Oo PR 4 wee aoe aes aoe - stuimoy SA ca 1.1 Z.0 LS Z.0 Sur g.1 ire eee 1piusou *) Qelt 4 9.0 Pa WA 2.9 1.b1 oe “* seppungsaqus “T OT o£ gti “ed t.o Z.0 Z.0 Fd as 1ygasing "7 2.0 0.0 0.0 ares 1.71 $.S tz Lz rep . “pup 'y t.9z L.oz 9.gh OL 6.54 Z.QI 4.9 SRS ee [etsy 9. S.11 L.E1 S.dz £6 9.8 S.1 “+ poasapoisig “y 00S 4g $28 00% Loz goor osh “ pauruexa ‘oN sdoo1y, |sivak gZ~Z1 uaIpytyD sivak og-1 | syoog | sxouostg | uostwg | squateg | szns9oy | puL sy[npy| sivad z1-0 Sody tv | oanen aaneny | asaqeyy | wnypdsy Away sqyuaeg | uerpytyo SYD ANE yeatdsopy (oz61) (9161) (oz6r) (6161) (eg161) (egr61) — (qg161) auoqsaydey]jsouu0g,O pur uodua A) Weyuog atu pure saoyayeyAy purjsuoane ra 4dhag Bae purpiugy Al @1av.L . rewaieml 98 SUMMARY Five hundred persons living in Mandos were examined for intestinal protozoa with the results tabulated above. The percentage of E. histolytica cysts recorded was somewhat higher than those reported from other countries for which figures are available, excepting Malta. I am indebted to Dr. H. W. Thomas for allowing me to make use of the material collected for hookworm examination for this investigation. REFERENCES BentuaM, T. (1920). Parasitology. Vol. XII, p. 72. Dosett, C., and Jerrs, M. W. (1917). Brit. Med. Fourn., May 12, p. 610. James, W. M. (1914). Ann. Trop. Med. & Parasit., Vol. VIII, p. 133. Kororp, C. A., Kornuauser, S. I., and Prats, J. T. (1919). ‘Fourn. Amer. Med. Assoc., Vol. LXXII, p- 1721. Mackinnon, D. L. (1918). Lancet, Vol. CXCV, p. 386. Maptesrone, P. A. (1920). Ann. Trop. Med. & Parasit., Vol. XIV, p.283. ss Mattuews, J. R. (1918). Ann. Trop. Med. & Parasit., Vol. XII, p. 17 Marttuews, J. R., and Smitu, A. M. (1918a). Ann. Trop. Med. & Parasit., Vol. XII, p. 349. (1918b). Anz. Trop. Med. G& Parasit., Vol. XU, p. 361. ——-— (1919). Ann. Trop. Med. & Parasit., Vol, XIII, p. 91. Soitn, A. M. (1918). Ann. Trop. Med. & Parasit., Vol. XII, p. 27. ——— (1919). Ann. Trop. Med. & Parasit., Vo). XII, p. t. Wenyon, C. M., and O’Connor, F. W. (1917). ‘Fourn. R.A.M.C., Vol. XXVIII, p. 1. Wooncocx, H. M., and Penrotn, W. J. (1916). Brit. Med. Fourn., Maxch 18, p. 407. 99 A PARASITE RESEMBLING PLASMODIUM FALCIPARUM IN A CHIMPANZEE BY B. BLACKLOCK AND S. ADLER From the Sir Alfred Lewis Jones Research Laboratory, Freetown, Sierra Leone (Received for publication 20 March, 1922) PLATE IX The following observations were made by us on a chimpanzee, Anthropopithecus troglodytes, at Freetown, Sierra Leone. The animal, according to the statement of the owner, had suffered from an attack of dysentery lasting from January ist to January 16th, 1922. It was examined by us on the 11th and 12th of January, at which time it was passing blood, pus and mucus. The only organisms found in the faeces by microscopical examination were large numbers of Blastocystis resembling Blastocystis hominis; no bacteriological examination of the faeces was made; the cutaneous blood was examined, and no parasites were found. From January 16th to January 31st the animal was well. On January 31st and February Ist it refused food ; its condition improved on the 2nd and 3rd, but the next day it became worse, and the owner handed it over to us. The chimpanzee was very thin, its hair was coming out and it was obviously ill; on February 5th it had an attack of diarrhoea; no blood was passed; Strongyloides larvae were present in large numbers in the faeces. Malaria parasites were found in the blood on February 4th. They increased in number and the animal’s condition became worse. On the 8th of February, in the afternoon, it was somnolent, and on the oth it refused food and drink, lay motionless, was not easily 100 roused, and remained in any attitude in which it was placed without attempting to change it. At 8.30 a.m., after considerable retching, it vomited. As the animal’s condition was grave and appeared to be associated with the increasing number of parasites in its blood, 5 grs. of quinine bisulphate in solution were administered orally at 10.30 a.m. From February rith till 19th its condition improved ; the number of parasites in the peripheral blood was reduced rapidly by the action of the quinine, but the blood was never free. On the 19th the blood was again heavily infected, more so than on any previous occasion; the animal was ill, but its condition was not as grave as it was on the oth, and by the 21st its appetite returned and it began to recover without any quinine. On the 22nd the animal was lively and eating well, and the parasites in its peripheral blood were decreasing. On February 23rd, at 8.30 a.m., the animal appeared well and made a good meal. At noon the same day it was found lying in its cage in a condition of collapse and breathing with difficulty ; it had vomited a large quantity of bile-stained material. Death occurred in half an hour. Post-mortem examination. The immediate cause of death appeared to be innumerable small haemorrhages which were uniformly distributed over the whole surface of both lungs; these haemorrhages were very recent, and on examination proved to be caused by the presence of Strongyloid larvae. An account of the changes produced by the larvae and the sites in which they were found will be given in a future communication. The trachea contained a small quantity of regurgitated food, but this was not sufficient to cause obstruction. The vessels on the surface of the brain were dilated; there were no haemorrhages on the surface or in the substance of the brain; there was no meningitis. The spleen was not greatly enlarged; it was very dark in colour and somewhat harder than normal. The liver was dark and congested. The bone marrow was dark red. The kidneys and the heart appeared normal. EXAMINATION OF SMEARS AND SECTIONS OF THE ORGANS Bram. A few trophozoites and gametocytes were found; the capillaries were not blocked with parasites; pigment was present in small amount. Iol Spleen. Trophozoites and schizonts were found, but were not numerous; masses of pigment were present; there was considerable fibrosis. Liver. This contained pigment in very large amount; it occurred in granules and in coarse masses; some of the smaller granules were found in the liver cells. Bone marrow. Trophozoites and gametocytes were present, and coarse pigment was plentiful. Blood. rophozoites and gametocytes were present, but were not very numerous; very heavily pigmented leucocytes were common. TYPES OF PARASITE FOUND IN THE BLOOD 1. Large amoeboid trophozoites resembling P. vivax, in pale enlarged red cells. 2. Large heavy looking trophozoites more or less band-shaped and equatorial, coarsely pigmented, resembling P. malariae. 3. Trophozoites resembling small rings of P. falciparum. The red corpuscles were not enlarged and retained their colour. 4. Gametocytes were found, indistinguishable from those of P. falciparum ; they were never present in large numbers throughout the course of the disease. No schizonts were found in the blood. The P. vivax and P. malariae forms were scanty ; they were found on the 4th and 5th of February, but were not seen subsequently. After the 5th of February the parasites seen were invariably of the P. falciparum type; they showed a certain amount of pleomorphism, but this was not more notable than in the case of the human parasite. The pleomorphism consisted in the appearance of slightly amoeboid and ¢enue forms. Crescents appeared in largest numbers on the 12th and 13th of February, but even then were not numerous; no exflagellating forms were found. IDENTITY OF THE PARASITE The few parasites of the P. vivax type corresponded to P. inui, Halberstaedter and Prowazek, 1907, in that the host cell was enlarged and pale, and did not present Schiiffner’s dots. As they and the P. malariae forms were not found in the blood on or after 102 the 6th February, the conclusions drawn from the experiments detailed below cannot be considered strictly applicable to them. Reichenow (1920) records the discovery of parasites identical morphologically with P. falciparum in chimpanzees and _ gorillas. These parasites were always found by him in association with P. vivax and P. malariae forms. He concluded that anthropoid apes are as sure a source of danger to Europeans living in West Africa as are negroes. The conclusion of Reichenow as to the identity of the parasite he found in gorillas and chimpanzees with the human parasite is interesting. The establishment of this identity would necessarily lead to important inferences. It would mean that anophelines which had fed on infected anthropoid apes could acquire salivary gland infection. Such anophelines, in parts remote from human habitation, would be capable of infecting any human being who came within their range. In this way they would constitute a permanent danger to persons employed in opening up new areas. Reichenow’s conclusion appears to us too far-reaching in view of the fact that it is based on morphological grounds only. If his conclusion is correct, it becomes difficult to understand why inocula- tions from human beings infected with malaria into chimpanzees should fail. The only successful inoculation of malaria from a human being into an animal is that performed by Mesnil and Roubaud (1920). These authors succeeded after several attempts in inducing a transient infection with P. falczparum in one of two chimpanzees. The incubation period was ten days, and the animal recovered spontaneously after another ten days. It is significant that in the two experiments using as vector A. maculipennis which had been infected from a case of P. falcifarum, transmission to the chimpanzees failed entirely. This alone would suggest that P. falciparum is not easily transmissible to the chimpanzee, in view of the ease with which infective anophelines transmit P. falciparum to human beings in laboratory experiments. The failure of Mesnil and Roubaud to transmit malaria to chimpanzees by the bite of infected anophelines raises the question as to whether the P. falciparum forms observed by them in the chimpanzee were really due to the inoculation or were a relapse of the parasite which occurs naturally in the chimpanzee. 103 In order to determine whether the parasite resembling P. falciparum found by us in the chimpanzee was capable of infecting human beings, we performed the following experiments. EXPERIMENTS WITH LABORATORY-BRED A. COSTALIS Laboratory-bred A. costalis were allowed to feed on the chimpanzee on two successive nights. After a lapse of from four to fourteen days from the first feed, forty mosquitoes were dissected, and in no case was infection found either in the gut or salivary glands. EXPERIMENTS WITH INJECTIONS OF INFECTED BLOOD Two Europeans were given subcutaneous and intravenous injections of blood from the chimpanzee. The first subject had never had malaria, and had taken prophylactic doses of five grains of quinine bisulphate daily from January 1oth to February 6th, 1922. The last dose was taken at 7 a.m. on February 6th. On February the 7th, at 5 p.m., he received subcutaneously 1 c.c. of the blood of the infected animal. An hour later slight nausea ensued, which lasted two hours. The local reaction was slight. On the goth of February, at 10 a.m., the same subject received an injection of 0°4 c.c. of the animal’s blood into his right median basilic vein. At this time the animal’s infection was heavy, 2z.e., four rings to the field (Obj. 1/12, Oc. 0, Leitz). Slight nausea followed a quarter of an hour after the injection, and lasted a few hours. The subject’s blood was examined twice daily from the date of the first injection, but no parasites were found. During an observation period of twenty-eight days, no infection occurred. An interesting fact was observed, namely, that from the 12th to the 14th of February transient urticarial patches occurred, localised round the site of the first inoculation. These patches appeared and disappeared several times during the course of the day. The second subject had previously suffered from malaria, and recently, within a year, from a P. falciparum infection, but had been free from relapse during the last six months. He was taking two grains of quinine bihydrochloride daily until the 7th February. 104 He received on February 19th, at 7 p.m., I c.c. of the animal’s blood subcutaneously; at this time the animal’s blood showed as many as nine rings toa field. No local or general reaction followed. On the 20th February, at 5 30 p.m., he received o'2 c.c. of the animal's blood intravenously. Examination of the subject’s blood before the first inoculation was negative, as were also subsequent examinations. No infection occurred during an observation period of seventeen days after the second inoculation. The results of the above experiments lend themselves to two explanations, viz., that the parasite is P. falczparum which has lost its infectivity for man by passage through the chimpanzee, or that it belongs to a new species of the genus Plasmodium. In view of the limited number of experiments performed, we consider it premature at present to decide definitely between these two interpretations. Our experiments so far certainly do not confirm Reichenow’s conclusion that chimpanzees as reservoirs of P. falciparum are a source of danger to Europeans in West Africa. SUMMARY A parasite morphologically indistinguishable from P. falczparum was found by us occurring naturally in a chimpanzee in Freetown, West Africa. This parasite appears to be the same as that described by Reichenow in chimpanzees and gorillas, and stated by him to be the human parasite. Laboratory-bred A. costalis fed on this chimpanzee failed to become infected, but, as stated above, crescents were few and exflagellation was not observed. We have failed to transmit the infection to two human subjects by subcutaneous and intravenous inoculation. REFERENCES Rercuenow, E. (1920). Ueber das Vorkommen der Malariaparasiten des Menschen bei den Africanishen Menschenaffen. Cent. f. Bakt. I. Abt. Orig. Vol. LXXXV, No. 3, pp: 207-216, ext. Trop. Dis. Bull., Feb. 1921. Mesniz, F., and Rousaup, E. (1920). Essais d’inoculation du paludisme au chimpanzeé. Ann. Inst. Past. Vol. XXXIV, No. 7, pp. 466-480, ext. Trop. Dis. Bull., Feb., 1921. jo™ inde > . isteegrerttha quit Yo ty = VS tees ae J 4 —* ; road} 7 fe 7 ’ \ ra ar 1 <= VBI ih wad t d an ‘ > oo ey aA He? 4 >) i Wie pT M4 Atel ad 5 7 7s x a cant). tity ‘rn / tit - } i7—* t ‘ ' aber), teh ; { c. 185. ¢ vill and ¢ ix—tergites of eighth and ninth segments; s viii and s ix—sternites of eighth and ninth segments; c—cerci; v.p.—ventral process of the tenth segment. concavity is anterior and is expanded laterally, and a more delicate bar of chitin in front of it, which is broadest in the middle line. Cerci prominent, short and broad, hollowed out on their inner aspects, and inserted obliquely, that is with their broad surfaces converging dorsally; in the ten specimens measured the length 168 ranged from 152 to 198m, average 183, and the breadth from 77” to 103, average $5. Ventral process of the tenth segment (fig. 7 @) with a moderately well developed notch in the middle of its posterior border, and bearing on each side several (about eight) long setae. Spermathecae three, highly chitinised, sub-spherical, the length being usually slightly greater than the breadth. The spermathecae are unequal and somewhat variable in size; the right and left ones are approximately the same size and in the ten specimens measured ranged in length from 61 to 80, average 694, and in breadth from 50” to 76”, average 63; the middle one is larger and ranged in length from 84 to 95m, average QIm, and in breadth from 70 to Qin, average 82u. A few pale spots similar to those in Anopheles costalis are sometimes present round the base. The ducts of the spermathecae are scarcely at all chitinised, at most for a distance of 24 to 4m at their commencement. La Fic. 7. Ventral process of the tenth abdominal segment, ventral view, of a—Stegomyia fasctata, F.; b—S. unilineata, Theo.; c—S. vittata, Bigot., and d—Ochlerotatus albocephalus, Theo. X c. 250. S. apicoargentca, {heo. One specimen. Very closely resembling S. fasciata. In the single specimen examined the only differences noted were that the setae on the posterior angles of the ninth tergite appeared to be longer, that the ventral process of the tenth segment was less deeply notched, and that the spermathecae were rather large, measuring 84m by 80n, 110% by gif, and 80n by 76x. These slight differences may be merely variations, and without confirmation from more materials are insufficient to distinguish the species. S. dendiophila, Edw. Two specimens. Apparently indis- tinguishable from S. /asczata. S. luteocephala, Newst. Four specimens. Very similar to S. fasciata, the only difference noted being the absence of the notch in the posterior border of the ventral process of the tenth segment. 169 S. metallica, Edw. One specimen. Apparently indistinguish- able from S. fasciaéa, but notch in posterior border of the ventral process of the tenth segment shallow, as in S. unzlineata. S. simpsoni, Theo. One specimen. Apparently indistinguish- able from S. fascia¢a, but the notch in the posterior border of the ventral process of the tenth segment is very shallow in the single specimen examined. S. unilineata, Theo. Six specimens. Apparently indistinguish- able from S. fascia¢a, but the notch in the posterior border of the ventral process of the tenth segment very shallow (fig. 7 0), as in S. simpsoni but cerci shorter. S. vittata, Bigot. Three specimens. Very similar to S. fasczata, but the ventral precess of the tenth segment long, tongue-like, without a notch (fig. 7 ¢). Genus Ochlerotatus Five species were examined; in all of them the genitalia were somewhat of the same type as those of species of Stegomyia. Two of the species, however, showed a remarkable divergence, inasmuch as they possessed only a single, large, spermatheca. O. albocephalus, Theo. Two specimens. Similar to S. fasczata, but eighth segment usually more or less retracted within the seventh, eighth sternite more widely notched, cerci rather longer and narrower, length about 1904, breadth about 63s, ninth tergite smaller, less highly chitinised, notch in the posterior border of the ventral process of the tenth segment much deeper (fig. 7d), and chitinised portion of the ducts of the three spermathecae a little longer, about 4 to 7H. : O. apicoannulatus, Edw. One specimen. Similar to O. albo- cephalus, but cerci relatively shorter and broader, length 134,p, breadth 65 in the single specimen examined, and spermathecac (especially the middle one) a little larger but still well within the range of variation found in S. /asciata. O. domesticus, Theo. One specimen. Similar to O. albo- cephalus, but eighth sternite more deeply notched, cerci longer, length about 2804, breadth about 68, and notch in the ventral process of the tenth segment less deep. 170 O. irritans, Theo. (fig. 8). Seven specimens. General characters similar to those of O. albocephalus. Eighth segment usually partially withdrawn within the seventh, but capable of complete protrusion, disclosing a wide membranous junction between the two segments. Eighth sternite shghtly longer than the tergite, notch rather shallow. Cerci of usual form: average length 186, average breadth 83,. Fic. 8. Ochlerotatus irritans, Theo., posterior extremity of abdomen of female, lateral view. X c. 185. Ventral process of the tenth segment deeply notched posteriorly, as in O. albocephalus. Spermatheca single, large, sub-spherical ; average length 93”, breadth 89”. There are at the base a number of pale spots, as in A. coséalis. Duct chitinised for only a short distance, about 5m, at its commencement. 171 O. punctothoracis, Theo. One specimen. Apparently almost indistinguishable from QO. ér7vitans, but the cerci are rather smaller in the single specimen examined and are more pointed at their tips. Genus Mansonioides M. africanus, Theo. (figs. 9 and 10). Eight specimens. Posterior extremity of the abdomen bluntly conical. Eighth segment may be partially retracted within the seventh, tergite narrow, posterior margin armed with a row of strong recurved teeth arranged as shown in the figure (fig. 9), the middle group composed of seven, or more commonly nine teeth, the central one being the longest, the two lateral groups of from five to seven teeth; sternite much longer than the tergite, and prolonged on each side posteriorly as a wide flap which is deeply notched. Ninth segment much reduced, the tergite represented by a narrow arch of chitin, and the sternite by the usual transverse sclerites, which are rather poorly developed. Cerci rather short with their narrowest diameter directed dorso- ventrally, slightly concave dorsally, and ending in a rather sharp tip; length variable, average about 192m, middle lateral breadth about 90”. Ventral process of the tenth segment very deeply cleft in the middle line posteriorly. Spermathecae three, two large and very highly chitinised, and one very small and feebly chitinised. The two large spermathecae are sub-equal and_ sub-spherical ; average length 137“, breadth 125m. They have a slight bulge near the point of origin of the duct (fig. 13 A), and there are numerous small pale spots at the base. The ducts are chitinised for only a short distance (about 10”) at the commencement. The small spermatheca is sub-spherical, length about 29, breadth about 28, it is usually feebly and incompletely chitinised, its base being membranous, and is difficult to find if the abdomen is incompletely cleared, and may for this reason be overlooked. Its duct joins the duct of one of the large spermathecae, so that it clearly represents an ill-developed Jateral spermatheca. M. uniformis, Theo. Five specimens. As in WM. africanus, but the lateral flaps of the eighth sternite are not notched. In the five specimens examined it was also noted that the teeth in the lateral groups on the dorsum of the eighth segment were rather more variable and numbered from three to six, and that the small } pee * aca yr Fic. 9. Mansonioides africanus, Theo., posterior extremity of abdomen of female, dorsal view. X ¢. 150. Fic. 10. Mansonioides africanus, Theo., posterior extremity of abdomen of female, lateral view. X c. 150. 173 spermatheca was sometimes rather large, in one instance measuring 65 by 53m; these latter differences are probably not specific. Genus Aedomyia Aedo. africana, Nev.-Lem. (figs. 11 and 12). Two specimens. Similar to Anopheles. Posterior extremity of the abdomen blunt, Fic. 11. Aedomyia africanus, N. L., posterior extremity of abdomen of female, dorsal view. X c. 185. Fic. 12. Aedomyia africanus, N. L., posterior extremity of abdomen of female, lateral view. xX c. 185. P. 173. Legends beneath figs. 1 and 12, for Aedomyia africanus read Aedomyia africana. 174 cerci not prominent. Eighth segment not withdrawn within the seventh; sternite with a shallow notch. Ninth segment much reduced, feebly chitinised. Cerci short and broad, with blunt, rounded ends; length 118, breadth 65. Ventral process of the tenth segment short and broad with a wide notch in its posterior border, bearing on each side several stout setae. Spermatheca single, very highly chitinised, resembling that of A. funestus ; length 106“, breadth 97, length of the chitinised portion of the duct 45,4 ; the whole spermatheca is sparsely dotted with pale spots, which, however, are small and are most numerous at the base. Genus Taeniorhynchus T. aurites, Theo. (fig. 13, B toD). One specimen. In some respects similar to Mansonioides. Eighth segment only slightly withdrawn from the seventh and capable of complete protrusion; sternite long, not notched, tergite short, without teeth. Ninth segment reduced, much as in Mansonioides. Cerci (fig. 13 Band C) D A B Fic. 13. d—Mansonioides africanus, Theo., spermatheca; x c. 375. B—Taentorhynchus aurites, Theo., outline of one of the cerci in ventral view, and C—in lateral view; and of D—ventral process of the tenth segment; X c. 185. curved dorsally, short, broad, with rounded extremities; length 1824, breadth Sou. Ventral process of the tenth segment hardly at all notched in the middle line posteriorly (fg. 13D). Spermathecae three, rather poorly chitinised, sub-spherical, large, unequal; in the single specimen examined they were not fully expanded, but, so far 175 as could be judged, their diameters were respectively about roop, 115m, and 122u. A short portion of the commencement of the ducts is feebly chitinised. Genus Culex Twelve species were examined, all of which possess genitalia of a very similar form, so that points of distinction, when found, are but slight and sometimes difficult to detect. In all the species the posterior extremity of the abdomen is blunt, the eighth sternite notched posteriorly, and the cerci relatively small, short, broad, and obliquely set. On the lining membrane, just below the posterior border of the eighth sternite, is a tuft-like group of more or less stout setae. The U-shaped structure surrounding the vulva is well chitinised. The ventral process of the tenth segment is short, occasionally notched, and not very hairy. There are three spermathecae, which are usually oval, and their ducts are chitinised for only a short distance. Points of distinction between. species appeared to be furnished by all the above structures. It may be mentioned here that the species belonging to the Genera Culiciomyia, Eumelanomyia, and Muicraedes, which we have examined, also possess genitalia of the same type. C. fatzgans, Wied. (figs. 14 and 15). Tenspecimens. Posterior extremity of the abdomen blunt, cerci not very prominent. Eighth segment not withdrawn within the seventh, sternite prolonged posteriorly beyond the tergite, and shallowly notched. From the middle of the membrane lining the posterior border of the eighth sternite arises a tuft-like group of about ten rather stout setae. Ninth segment, as usual, much reduced; tergite a narrow strip, broadest laterally and rather feebly chitinised. Ventrally there is a horseshoe-shaped strip of chitin, open anteriorly, enclosing the vulva, and just posterior to it a wider arch of chitin, from the lateral portions of which rather broad but feebly chitinised plates project inwards. Cerci set slightly obliquely, concave internally, short and broad, with truncated ends; in the ten specimens measured, length from 137m to 170m, average 150m, and breadth from 80,4 to giz, average 85. Ventral process of the tenth segment (fig. 17 6) without a notch, bearing at its apex a few stout setae, and on the ventral aspect a few (two or three on each side) smaller ones. 176 Fic. 14. Culex fatigans, Wied., posterior extremity of abdomen of female, dorsal view. X c. 185. Fic. 15. Culex fatigans, Wied., posterior extremity of abdomen of female, lateral view. X c. 185. 177 Spermathecae (fig. 16) three, very highly chitinised, sub-equal, the middle one being slightly the largest ; in the ten specimens measured the length ranged from 72m to 9g, average 84, and the breadth from 55 to 76m, average 634. They are somewhat variable in shape but are usually oval, sometimes almost sub-spherical, and commonly the base is rather broad and the apex narrowed so that they resemble a bee-hive. At the base there are a few ‘ pale spots.’ The chitinised portion of the ducts is short, conical, and in the specimens measured ranged in length from 6y to IIp, average 8p. O00 one Fic. 16. Outlines of spermathecae of a—Culex insignis, Cart. ; b—C. annulioris, Theo. ; c—C. fatigans, Wied.; d—C. consimilis, Newst., and e—C. duttoni, Theo.; and of f—the ventral process of the tenth segment, and g—the chitinous hoop round the vulva, of C. annulioris, Theo. All X c. 375. C. annulioris, Theo. One specimen. As in C. fatigans, but the inner chitinous bar enveloping the vulva (fig. 16g) is narrower, almost V-shaped, and the ventral process of the tenth segment 1s shallowly notched and bears four or five small setae on each side on its ventral aspect (fig. 17d). The spermathecae (fig. 166) are 178 highly chitinised, sub-equal, oval; length 99”, breadth 68, the chitinised portion of the ducts very short, about 2. C. consimilis, Newst. One specimen. As in C. fatzgans, but the spermathecae are rather larger (fig. 10d). They are highly chitinised, a rather long, oval shape, and not narrowed at the apex; in the specimen examined the middle one measured 137, in length by 84 in breadth, and the chitinised portion of its duct was about 7m long, and in the other two spermathecae the corresponding Se / VO eae oN F A NVA b c d Fic. 17. Ventral process of the tenth segment, ventral view of a—Culex consimilis; b—C. fatigans ; c—C. decens; d—C. annultoris; e—C. duttoni ; f—C. insignis; g—C. pruina; b—C. quasigelidus; i—C. rima; k—C. thalassius; I—C. tigripes var. fuscus; and m— C. tritaentorhynchus. X 250. measurements were 120“, 80, and 4 respectively. The ventral process of the tenth segment (fig. 17@) is more hairy than in C. fatigans, and bears about nine small setae on-each side on its ventral aspect. C. decens, Theo. Twelve specimens. Similar to C. fatigans, but in the specimens examined the cerci were rather more prominent, length about 160, breadth about 70,4 ; the tuft of setae on the lining 179 membrane of the posterior end of the eighth sternite usually rather larger, composed of about a dozen setae ; chitinous loop enclosing the vulva not so wide, more U-shaped; the ventral process of the tenth segment (fig. 17¢) with a shallow notch, small setae on ventral aspect rather variable, from three to nine on each side; and the spermathecae not so highly chitinised, a little larger, average length gop, breadth 68x. C. duttoni, Theo. Two specimens. Generally similar to C. fatigans. Tuft of setae on the lining membrane of the eighth sternite rather larger, composed of twelve setae; and ventral process of tenth segment (fig. 17) more hairy, bearing about six to nine small setae on each side on its ventral aspect. Cerci short and broad; length about 175, breadth about 115“. Spermathecae (fig. 16e) very highly chitinised, sub-spherical, the middle one slightly the largest; average length about 85, average breadth about 77, the chitinised portions of the ducts conical, rather long, average length about 15. C. insignis, Carter. One specimen. Generally similar to C. fatigans. Eighth sternite more deeply notched posteriorly ; and ventral process of the tenth segment (fig. 17 /) more conical and more hairy, bearing about six or seven small setae on each side on its ventral aspect. Chitinised bar encircling the vulva rather strong and thick, and omega-shaped. Tuft of setae on the lming membrane of the eighth sternite rather small, composed of eight setae. Cerci rather small and curved dorsally; length 114, breadth 57». Spermathecae (fig. 16@) very highly chitinised, oval, the middle one measuring about 95 by 68m, and the lateral ones 84 by 574; the chitinised portion of the ducts is short (5” to 6“) and narrow (44). C. pruina, Theo. One specimen. Apparently almost indis- tinguishable from C. /atigans, but in the single specimen examined the chitinous loop enclosing the vulva was rather narrower, as in C. annulioris, and the spermathecae were not so heavily chitinised, more regularly oval, and longer, having a length of about 105z, breadth about 80, and the chitinised portion of the ducts about 6p. Small setae on the ventral aspect of the ventral process of the’ tenth segment (fig. 17g) rather more numerous, about six or seven on each side. 180 C. quasigelidus, Theo. One specimen. As in C. fatigans, but in the single specimen examined the loop of chitin enclosing the vulva is more V-shaped, and the spermathecae are rather larger, the middle one measuring 103# by 722, the lateral ones g1# by 69p, and the ducts being short, 4“ and 2. respectively. C. vima, Theo. One specimen. Closely resembling C. zzszgnis. In the single specimen examined the cerci were small and curved dorsally, as in C. zzsignis, and the apices of the spermathecae were broad and not narrowed as they often are in C. fatzgans. The eighth sternite also appeared to be more deeply notched than in C. fatigans, and the ventral process of the tenth segment (fig. 17 7) more hairy, bearing a row of about five small setae on each side on the ventral aspect. C. thalassius, Theo. Seven specimens. Similar to C. fatzgans, but loop of chitin enclosing the vulva rather narrower posteriorly, and spermathecae more regularly oval, and in some specimens a little larger (103# by 68 in one). Small setae on the ventral aspect of the ventral process of the tenth segment (fig. 17) rather more numerous, about five on each side. C. tigripes, Grp., var. fuscus, Theo. Three specimens. Very highly chitinised. Generally similar to C. fatzgans, but larger. Cerci about 180” by 105m. Spermathecae very highly chitinised, shaped as in C. fategans, the middle one the larger, about 110” by 85, the lateral ones about 97” by 76; the chitinised portion of the ducts is about 84 long. The tuft of setae on the lining membrane of the eighth sternite is rather larger than in C. fatzgans. The ventral process of the tenth segment (fig. 17 Z) is not notched, and is rather more conical and hairy than in C. fatigans, there being about six small setae on each side on the ventral aspect. C. tritaeniorhynchus, Giles. Two specimens. Apparently indis- tinguishable from C. /atigans, but the spermathecae are, perhaps, a little more regularly oval, and the ventral process of the tenth segment (fig. 17 #) rather more hairy, having four or five small setae on each side on its ventral aspect. The genitalia of the twelve species examined were so much alike that they could be distinguished, if at all, only by means of minute differences, which in some cases cannot be accepted as of specific value owing to the materials being insufficient to exclude the error 181 due to the natural range of variation. . Judging solely from the specimens we have examined, however, points of distinction appeared to be present in the size of the cerci, the shape and size of the spermathecae, the shape of the chitinised hoop round the vulva, the shape of the ventral process of the tenth segment, and the number of small setae (not including the larger setae near the apex) on the ventral aspect of this process. From all the other species examined C. duttoni is readily distinguished by the sub-spherical shape of the spermathecae and the relatively long chitinised portion of the ducts, and C. consimilis by the large size of its oval spermathecae and the relatively numerous small setae (about nine on each side) on the ventral aspect of the ventral process of the tenth segment. Two other species, C. annulioris and C. decens, may, perhaps, be separated by the fact that in them the ventral process of the tenth segment is notched; and C. tigripes may be recognised by. its size. Other points that may be of systematic value, such as the small size of the cerci in C. rima and C. imsignis, and the scantiness of the hairs on the ventral process of the tenth segment in C. fatigans and C. quasi- gelidus, can be confirmed only by further experience. Genus Culiciomyia C. nebulosa, Theo. Five specimens. Genitalia of the same type as in the genus Culex, and very similar to those of C. fatigans, from which they appeared to differ only in having shorter cerci (about 115m by 72), and in the ventral process of the tenth segment being shallowly notched and slightly more hairy, having about five small setae on each side on the ventral aspect. The spermathecae are also rather larger and less highly chitinised; the average measurements of the middle one being, length 994, breadth 69 (one specimen measuring 110“ by 72,), and the lateral ones, length 85, breadth 63; the chitinised portions of the ducts measure about 7» in length. Genus Eumelanomyia E. inconspicuosa, Theo. Three specimens. Genitalia of the same type as in the genus Culex. Posterior extremity of the abdomen blunt, cerci not prominent, seldom projecting beyond the 182 eighth sternite. Tuft on the lining membrane of the eighth sternite small, composed of eight not very strong setae. Ninth segment as in C. fatigans, but loop enclosing the vulva very feebly chitinised. Cerci small, broad, extremities directed inwards; length about 105, greatest lateral breadth about 50@. Ventral process of the tenth segment not notched, bearing a few hairs, none of which are very strong. Spermathecae three, oval, rather feebly chitinised, the middle one slightly the largest; average length about 88, breadth 65, the chitinised portion of the ducts short, about 7x. Genus Jlicraedes M. inconspicuosus, Theo. One specimen. Genitalia of the Culex type. Posterior extremity of the abdomen blunt, cerci not prominent. Eighth segment not retracted within the seventh; sternite with a shallow notch posteriorly. Ninth segment reduced as usual. Ventrally there is a small U-shaped bar of chitin enclosing the vulva, and more posteriorly a second strip of chitin forming a transverse arch. From the lining of the posterior part of the eight sternite, in the middle line, there projects backwards a tuft-like group of eight (four pairs) stout setae. Cerci short and broad, with truncated extremities; length 76, breadth 45z. Ventral process of the tenth segment projecting a little beyond the cerci, broad, very slightly notched, and bearing on each side a few rather feebly chitinised setae, two of which, one apical and one slightly dorsal, are rather large. Spermathecae three, relatively large, moderately well chitinised, oval, and sub-equal; length about 57m, breadth about 46”. The chitinised commencements of the ducts are conical and rather long, about 104. Genus Mzmomyza M. splendens, Theo. (fig. 18, A and B). Three specimens. Posterior extremity of the abdomen bluntly conical, cerci rather prominent. Eighth segment not withdrawn within the seventh. Ninth segment much as usual; no U-shaped loop of chitin enclosing the vulva. Cerci of the usual form, obliquely set, rather small, length about 115, lateral breadth about 65. Ventral process of the tenth segment reaching posteriorly as far as the cerci, broad, very 183 hairy, apically and ventrally, and deeply notched. Spermatheca single, highly chitinised, sub-spherical, and relatively very large, diameter about 105m to 115; there are numerous pale spots at the base, and almost no part of the duct is chitinised. M. mimomyiaformis, Newst. Two specimens. Similar to M. splendens, but in the specimens examined the cerci were very short and broad, length 95, lateral breadth 72m, and the ventral process of the tenth segment projected posteriorly beyond the cerci and was only feebly notched (fig. 18 Cc). — —/-svii Sem bd ee Fic. 18. Mimomyia splendens, Theo., posterior extremity. of abdomen of female. A— ventral view; B—lateral view. X c. 185. Mimomyia mimomyiaformis, Newst.; C—ventral process of the tenth segment, ventral view. X c. 185. M. plumosa, Theo. (fig. 19). One specimen. Genitalia unlike those of the two preceding species. Chitinisation of the ninth sternite rather strong, but there is no loop enclosing the vulva. Cerci obliquely set as usual, appearance varying greatly with the position: in a ventral view they are truncated, in a lateral view they are cone- shaped with a rather pointed extremity, and in sub-lateral view (the lateral aspect of the cerci) they are short and broad, about 150# by 115m, with their dorsal extremities prolonged into a 184 process. Ventral process of the tenth segment large, very hairy, deeply notched posteriorly (ig. 19B). Spermathecae three, highly chitinised, sub-spherical to oval, the middle one the largest and measuring about 148 by 1374, the lateral ones smaller, about 1222 by 1064; practically no part of the ducts is chitinised. B Gc Fic. 19. Mimomyia plumosa, Theo. A—posterior extremity of abdomen of female, lateral view ; B—ventral process of the tenth segment, ventral view ; and C—one of the cerci, gub-lateral view. X c. 185. Genus Uvanotaenia U. balfouri, Theo. One specimen. Very small, posterior extremity of the abdcmen bluntly conical, the terminal segments not so far retracted as usual. Cerci (fig. 20 B) very short, broad; length 185 about 60n, breadth about 45. Ventral process of the tenth segment about as long as the cerci, broad, without a notch and bearing a few but no very large setae. Spermatheca single, sub- Fic. 20. A—Uranotaenia annulata, Theo., posterior extremity of abdomen of female, lateral view. X c.185. B—Uranotaenia balfouri, Theo., one of the cerci, lateral view. Xc. 185. spherical, length abcut gia, breadth about 84; only the very commencement of the duct is chitinised. U. annulata, Theo. (fig. 20A). One specimen. Generally similar to U. balfouri, but larger. Cerci rather long, with bluntish ends; length about 1704, breadth about 68”. Ventral process of the tenth segment nearly as long as the cerci, without a notch, bearing numerous setae, those at the apex being large. Spermatheca single, sub-spherical, length Son, breadth 76m; practically no part of the duct (which is long and narrow) 1s chitinised. Tribe SABETHINI Genus Eretmopodites E. chrysogaster, Grah. (figs. 21 and 22). Six specimens. Very highly chitinised. Posterior extremity of the abdomen blunt, cerci Fic. 21. Eretmopodites chrysogaster, Grah., posterior extremity of abdomen of female, dorsal view. X c. 185; Fic. 22. Eretmopodites chrysogaster, Grah., posterior extremity of abdomen of female, lateral view. X c. 185. 187 projecting slightly. Eighth segment not withdrawn within the seventh, sternite projecting slightly further back than the tergite and with its posterior margin deeply notched in the middle. Ninth segment as usual much reduced, chitinised plates rather strong, arranged as shown in the figure. Cerci with blunt or truncated ends; average length about 200, breadth about 100. Ventral process of the tenth segment shorter than the cerci, deeply notched, bearing on each side numerous setae, one pair very strong. Spermathecae three, highly chitinised, sub-spherical; the middle one is the largest and has a diameter of about 110 or more, the lateral ones are a little smaller, and are usually, but not always sub-equal, and have a diameter which in the specimens examined ranged from QIm to 114m, average 97m. The commencement of the ducts is chitinised for only a short distance, about 6; and there are a few pale spots round it at the base of the spermathecae. E. quinquevittatus, Theo. (fig. 23). One specimen. Apparently Fic. 23. Eretmopodites quinquevittatus, Theo., posterior extremity of abdomen of female, lateral view. X c. 185. 188 almost indistinguishable from Z. chrysogaster; but in the specimen examined the cerci were slightly smaller, about 170” by 87, and so were the spermathecae, the diameters of which were about 724, QI, and 68, respectively, and no part of the ducts appeared to be chitinised. 189 NOTES ON AUSTRALIAN CESTODES BY P. Ay MAPLESTONE AND Lf. SOUTHWELL (Receiwed tor publication 19 May, 1922) V. THREE CESTODES FROM THE BLACK SWAN The three following species of Cestodes were found in the intestine of Chenopsis atrata, Lath. (the Black Swan), several of which were examined at Townsville, North Queensland :— (1) Nematoparataenia paradoxa, n. g., N. sp. (2) Echinorhynchotaenia nana, n. sp. (3) Hymenole pis lanceolata (Bloch, 1782), Weinland, 1858. (1) Nematoparataenia paradoxa, n. g., n. sp. On a single occasion about twenty specimens of this worm were obtained. EXTERNAL ANATOMY. The worm measures about 9 mm. in length and 4 mm. in breadth except at the posterior extremity, where it expands into an oval saccular portion measuring about o'8 mm. in length by 06 mm. in breadth. . The cuticle exhibits no trace of segmentation, even under high magnification. In cross-section the worm is circular with a ventral indentation (figs. 3 and 4). Head. ‘The head is armed with four suckers measuring 804 to 100m” in diameter. They are borne on short pedicles about 1004 long, which are situated about 200 from the anterior extremity. The anterior end of the head is occupied by a wide cup-shaped cavity about 400m deep, bearing round its margin twelve flattened tentacular processes with ‘minute spines about 2 long closely arranged around their borders. These tentacles are similar to those seen in the various species of the genus Parataenia, Linton; they 190 measure about 120” long and 40m broad. There is a well marked neck about 300% in length, which narrows to about 300 in Fic. 1. Nematoparataenia paradoxa, n.g., n.sp. Complete worm. a—position of fig. 2 ; b—position of fig. 3; c—position of fig. 4; m.c—mouth cavity; ov—ovary; ¢.p—tentacular processes; ut—uterus. X 17. . diameter. The remainder of the worm is cylindrical with a longitudinal groove running along its ventral surface (fig. 1). IgI INTERNAL ANATOMY. Muscular system. ‘This consists of a series of separate longi- tudinal fibres arranged in an irregular double row immediately beneath the cuticle. No transverse or dorso-ventral fibres were seen (figs. 2 and 3). Fic. 2. Nematoparataenia paradoxa n.g., n.sp. Transverse section at a—fig. 1; /.m.— longitudinal muscle fibres; ov—ovary; t—testes. % I40. Fic. 3. Nematoparataenia paradoxa n.g., n.sp. ‘Transverse section at b—fig. 1; /.m.— longitudinal muscle ; ov—ovary ; ut—uterus. X I40. Nervous and excretory systems. No details of these could be made out. 192 Genitaha. Testes. The testes are small and extremely numerous, they le in the dorsal and lateral fields (fig. 2); towards the middle of the worm they begin to become fewer in number. Vas deferens. No vas deferens, cirrus, or genital pore was seen. Ovary. he ovary is situated ventrally and occupies the middle threequarters of the worm’s length. In whole mounts the ovary shows no trace of segmentation, except that the lateral margins are serrated (fig. 1); in cross-section it is fan-shaped, the lobes radiating dorsally and laterally from a central point opposite the ventral groove; towards the posterior it gradually atrophies. Fic. 4. Nematoparataenia paradoxa n.g., n.sp. Transverse section at c—fig. 1; ov— ovary ; “t—uterus. X I4o. Vagina and receptaculum. ‘These structures were not seen. Uterus. The uterus begins about the junction of the middle and posterior thirds of the worm. It first appears at each side close under the cuticle, and as the ovary atrophies the two lateral limbs of the uterus gradually increase in size until they unite, and finally it occupies the whole of the body. Eggs. The eggs are circular and measure about 10” in diameter ; further details could not be determined. DIAGNOSIS. This worm resembles Pavataenta medusia, Linton (1899), only in its possession of tentacular processes on the head. It also bears a superficial resemblance to Nematotaenia dispar, Liihe (1899) in being unsegmented. Apart from these slight resemblances to the 193 above two species, this worm has characters entirely different from any known worm; this necessitates its being placed in a new genus, which we have named Nematoparataenia, and of which the following is the definition :— Nematoparataenia, n.g. Cylindrical worms with four suckers, and a number of digitate processes on the head. No trace of internal or external segmenta- tion.* Type species Nematoparataenia paradoxa. The type species is in the Museum of the Liverpool School of Tropical Medicine. (2) Echinorhynchotaenia nana, n. sp. About twenty specimens of this worm were obtained. Unfortu- nately the material was in very poor condition, so a full description is not possible. EXTERNAL ANATOMY. The largest worm measured about 2 cm. in length and including the cuticular expansions, which occur on the posterior borders of the segments, 1°77 mm. in breadth; the breadth of the worm without these expansions is about I°3 mm. Head. The head is about 1°5 mm. broad and 2°3 mm. long. Viewed anteriorly it is square, with rounded corners; each corner is occupied by a very strongly developed sucker looking almost directly forwards, and with a diameter of about 4504. In the centre of the anterior surface there is a small pit. When viewed from the side, the anterior surface is bluntly rounded, and the central pit, which is almost 300” deep, is seen to lie anterior to the suckers. Behind each sucker is a lappet, as in Axofplocephala perfoliata (Goeze, 1782), Blanchard, 1848. Behind the lappets the the head narrows gradually to a width of about 6004, at which point it is sharply marked off from the narrower anterior segments, which it tends to overlap, by a cuticular collar-like ring. There is no neck (fig. 5). * Although we were unable to see definite signs of segmentation, it should be noted that all our specimens were fully gravid and, therefore, old, and it is quite possible that in younger worms there would be segmentation in the internal organs. 194 Segments. ‘The segments-are broader than long, the most fully developed being 2 mm. broad and 200m long. They are like a number of saucers placed one within the other with the concavity facing posteriorly. This appearance is caused by the whole circumference of the posterior borders of the segments being provided with cuticular expansions about three times as long as the segments themselves. In cross-section the segments of the anterior two-thirds of the worm are nearly circular, whilst those of the posterior third are oval. The genital pores are unilateral and open on the right side. Fic. 5. Echinorhynchotaenia nana, n.sp. Scolex. /—lappet ; p—fragment of proboscis ; s—beginning of strobila. X 17. INTERNAL ANATOMY. Muscular system. The longitudinal muscle is disposed in two layers, an outer feebly developed layer consisting of a few small bundles, and a relatively enormously developed inner layer measuring 300m in thickness (fig. 6). External to these are a few transverse fibres. No dorso-ventral fibres were seen. Four strands from the internal longitudinal layer run one to each sucker; the latter organs are extremely muscular, and in some specimens had actually fallen out of the scolex and appeared as almost spherical solid bodies. Nervous system. ‘There is a single lateral nerve on each side of the body lying external to the excretory vessels. Excretory system. A number of excretory tubes can be seen in the head, and these unite to form two lateral vessels on each side. The two lateral vessels are of about the same diameter, and one lies directly dorsal of the other. 195 Genitalia. Testes. The testes are three in number and they lie behind the ovary in the same transverse plane, two being on the aporal side. In full development they measure about 604 in diameter. Imi. |.m.2. ex. seat = me —SS= 5 Tir an 3 GRae A.M.B.,del. Fic. 6. Echinorbynchotaenia nana, n.sp. Transverse section towards posterior part of worm. ex.v.—excretory vessels; /.m.1.—outer longitudinal muscle layer; /.m.2.—inner longitudinal muscle layer. X 70. Vas deferens. ‘The vas deferens expands into a fairly large vesicula seminalis lying anterior to the mesial end of the cirrus pouch and connecting with the latter organ by a narrow duct. The cirrus pouch is 5004 long and 80u broad, extending almost half-way across the segment. The cirrus is as long as its pouch and ends in a club- shaped extremity, the extreme end of which is surrounded by a small sphincter muscle, The external surface of the cirrus 1s closely Fic. 7. Echinorbynchotaenia nana, n.sp. Termination of sex ducts. c—cirrus; ¢.p.— cirrus pouch; cu—cuticle ; g.a.—genital atrium; g.p.—genital pore ; r.m.—retractor muscle ; s.m.—sphincter muscle at tip of cirrus; v.—vagina; v.p.—vaginal plug. 4o. covered with minute spines. From the lateral border of the segment the male duct extends into the cuticle as a thin-walled tube, and it ends at its junction with the vagina which occurs about the centre of the cuticular expansion. From this junction the genital atrium runs laterally to open on the anterior surface of the cuticular prolongation about the junction of its inner and middle thirds (fig. 7). 196 Ovary. The material was in such a bad state of preservation that details relating to the ovary and vitelline glands could not be made out. ‘The ovary is centrally situated in front of the testes, and all that could be seen was a number of acini, each measuring about 30pm in diameter. Vagina and receptaculum. The vagina opens into the genital atrium immediately ventral to the male pore, and lying in its open end is a solid conical plug with a broad base (fig. 3). This plug is inserted into the slightly funnel-shaped opening of the vagina, and around the opening is a strongly developed muscle, which from the radial arrangement of its fibres probably acts as a retractor, drawing the walls of the vagina away from the plug. From the pore the vagina passes inwards anterior to the cirrus pouch, narrowing slowly until just internal to the excretory vessels it expands into a club- shaped receptaculum seminis, which runs as far as the median plane. Uterus. The uterus is a simple transverse sac loosely packed with eggs. Eggs. The eggs are circular and measure 40m in diameter, and the oncosphere measures 324. DIAGNOSIS. Fihrmann (1909) erected the genus LEchinorhynchotacnia to accommodate a species which possessed a proboscis-like rostellum armed with spines. Our worm closely resembles Fiihrmann’s species in its general anatomy except that the characteristic proboscis had been apparently torn out in all our specimens, but the appearance of the head, with a few ragged fibres protruding from the central pit, leaves no room for doubt that a proboscis has been present. The points in which our species differs from Fiihrmann’s £. ¢rz¢esticulata are the following :— E. tritesticulata E. nana, n.sp. Length ee 06 sie 4te otk 30 cm. 2cm. Breadth se tee oes eos oo 4-5 mm. °'7 mm. Lappets co: ane co oa ut absent present Apparatus at vaginal pore ... aa a absent present Genital atrium x0 a “8 ac absent present Position of genital pore... nc .-| On anterior of lateral | On anterior surface of border of segment. cuticular expansion 197 We, therefore, consider ours a new species, and name it Echinorhynchotaenia nana. The type specimen is in the Museum of the Liverpool School of Tropical Medicine. (3) Hymenole pis lanceolata (Bloch, 1782), Weinland, 1858. This cestode was found on four occasions. Many hundreds of specimens were obtained, and as they showed a wide variation in size and development, it is proposed to discuss these variations, since apparently they have not been noted in previous descriptions of the species. The largest specimen was 55 mm. in length with a maximum breadth of 5 mm., and from these dimensions there were worms of every gradation in size down to specimens only 11 mm. in length by o°3 mm. in breadth; that this difference in size is not altogether due to different ages of the specimens is shown by the fact that many of the smallest worms had a fully gravid uterus in their posterior segments. Some of the smaller worms have only a part of the genitalia present. That is, either the male or female organs may be completely absent, but in no case was a worm seen in which both sets of glands were absent. In some without testes the uterus contains eggs; probably this is brought about by cross-copulation between different individuals. It may be held that the testes were originally present and have atrophied, but this is unlikely, as in the larger normal worms testes and ovaries are present together in all of the mature segments. In these small varieties the muscular system is poorly developed, with the result that the worms are very thin and diaphanous when compared with the larger ones. Another abnormality which was frequently observed was that the segments immediately behind the scolex rapidly increased in breadth in the normal manner, but after about the twentieth segment, instead of continuing to increase they becaine successively narrower for about a similar number of segments, after which the usual gradual and continuous increase took place. At first glance it would appear that worms of different species were included under the one head, but that this is not the case is shown by the fcllowing points :— 198 (1) The scolex and the few segments immediately following it are the same in all cases. (2) The cirrus, when present, is always of the same relative length and shape, no matter what the size of the worm. (3) The male and female genitalia occupy the same relative positions in the segments, whether present complete or only in part. (4) When a long series of material is examined, a regular sequence from the largest to the smallest worms can be obtained. As normally developed worms departed in no particular from previous descriptions of the species H. /anceolata, detailed anatomy has not been given. REFERENCES Fiinrmann, O. (1909). Die Cestoden der Vogel des weissen Nils. Results of the Swedish Expedition to Egypt and the White Nile, No. 27. Neuchatel. Lryton, E. (1899). Notes on Entozoa of Marine Fishes of New England, with Descriptions of several new species. U.S.d. Fish. Comm. Report for 1897. Line, M. (1899). Zur Anatomie and Systematik der Bothriocephaliden. M. Verh. Deutsch. Zool. Ges. Leipzig. 199 THE INCIDENCE OF A _ DISEASE IN POPULATION GROUPS, THE NUMBER OF PEOPLE IN WHICH IS KNOWN OR UNKNOWN BY Jae? WiseSTERHENS (Received for publication 25 May, 1922) As an example of the ‘ incidence,’ ‘ occurrence,’ or ‘ distribution’ of cases of a disease in one or more groups, such as age-groups of a population, the number of people in which is uxknown, we may take the following. Of a total of twenty cases of influenza, let us suppose that ten occurred in Group A and ten in Group B, then the respective incidences ten and ten are equal, and the number that occur in each group per one hundred cases, viz., fifty and fifty, are also equal. As an example of the ‘ incidence,’ ‘ occurrence’ or ‘ distribution’ in age-groups, the number of people in which is known, we may take the following. Of a total of twenty cases of influenza, let us suppose that ten occurred in Group A, containing one hundred people, and ten in Group B, containing fifty people, then the incidences are IO per cent. and 20 per cent. respectively (and the ratios of the incidences per one hundred cases 33 per cent. and 66 per cent. respectively). It will be evident that the term ‘incidence’ has been used here in two different senses. In the first sense of the term, ‘ incidence,’ it is only the number of cases that is known. In the second sense, when not only the number of cases but also the number of people among whom the cases occur is known, the term is applied to a figure expressing the number of cases that occur per one hundred people in each group. To emphasise the distinction in meaning between these two uses of the term ‘incidence,’ it would seem advisable to confine the term ‘incidence’ to the use of the term in the first sense, and the term ‘incidence rate’ to the use of the term in the second sense. 200 In practice, however, certain deductions are often made when the number of cases alone is known, which can, as we shall see, be only justifiably made when the number of people in the groups is also known, 7.e., when the ‘zzczdence rate’ can be calculated. In regard to ‘incidence,’ the larger the group, the larger (ceteris paribus) is the incidence. In regard to ‘incidence rates,’ the factor of unequal size of the groups, if it exists, is eliminated, as the rate is calculated for one hundred people in each group. The above examples may be tabulated as follows, using the words incidence and incidence rates in the sense defined above. Taste I. Shewing distinction between incidence and incidence rate. I | 2 3 4 5 6 7 | Incidence Total | Number Incidence, Number | rate, 7.c., Ratios of number | of people 1.¢., total of cases number Ratios of the of people that occur, number that occur! of cases the incidence Group that occur} in each of cases ineach | occurring incidence | rates to in each group | observed group = among rates to one group per 100 (that occur) per 100 | 100 people one another (Census) | people in each cases in each another | per cent. (Census) group | group | Ex. 1. A Co) 50 | B Io 5° Ex. 2 \ 100 66°6 10 50 Io I 33°3 B 50 33°3 Io 50 | 20 2 66°6 From the second example in the table we see that the ‘ liability to attack’ of a person in Group A 1s 10 per cent. and in Group B 20 per cent., z.e., it is twice as great in Group B as in Group A. This fact cannot, however, be deduced from the figures in the first example, because, although the number of cases is the same as in the second example, nothing is known as to the number of people among whom the cases occurred. It is the incidence rates (actual or relative) that are of importance if we are studying what may be termed the ‘real incidence’ of the disease on a group. 201 DIABETES We find recorded in Osler and Macrea, System of Medicine, second edition, p. 675, the age-group incidence of three hundred and thirty-five cases of diabetes in Baltimore (column 3) from which can be readily calculated the age-group incidences per one hundred cases (column 4). The figures for the age-group, distribution or incidence of the population of Baltimore per one hundred people are not given, so that for purposes of illustration I have used those of Liverpool as deduced from the 1911 census (Table II, column 2). Taste II. Showing incidences and ratios of incidence rates in Diabetes. | | 1 perp 13 so) | ee | 6 7 | | Incidence | | Total | Number | Incidence,) Number | rate, i.e., | Ratios of number | of people | i.e., total, of cases | number | Ratiosof | the of people | that occur; number (that occur, of cases | the | incidence Age-Group that occur} in each of cases | ineach | occurring incidence | rates to ineach | group observed group | among ratesto | one | group | per roo |that occur, per 100 | 100 people one | another | (Census) people | ineach | cases | in each another | per cent. (Census) | group | group I-10 ae oe 23°3 8 FAL Ws 070935 | 0°86 11-20 fea aay) 18"9 | 25 7°34. one 0°3883 3°59 } | 21-30 5 me 16°7 | 44 Taorieae tt o724s. | 6°71 31-40 ie ie 158 | 61 18'2 | re 11519 10°67 41-50 nes ae “ne | 6y 20°6 se 18392 | 17°04 51-60 23s se 73 «| 89 26°5 cess 3°6301 33°64 61-70 zee oo $5 33 g'8 | “55 21111 19°47 71-80 20 | 6 rey 0°8500 7°87 81- o°%3 ° ~ 10 | be oo foyfe) 100°0 335 99°42 | res 10°7886 99°85 If we were dealing with the total number of people in each group (column 1) instead of the number per one hundred of the population, and divided a figure in column 3 by the corresponding figure in column 1 and multiplied the result by one hundred, the figures obtained would represent the incidence rates, 7z.¢., the incidence per 202 one hundred people (column 5). But, in the present case, where we have divided the percentage figures in column 4 by the percentage figures in column 2, the resulting figures (column 6) represent simply the vatios* which the incidence rates bear to one another, and from these we can easily calculate the ratios, when the sum of the ratios 1s one hundred (column 7). Thus, to refer to Table II (column 7), we see that of one hundred cases of diabetes about thirty-three would occur among so many people in the age-group 51-60, while about half that number (17°04) would occur among the Same number of people in the age-group 41-50, whereas, considering the incidence merely (column 4), it 1s about the same in the two groups, viz., 20°6 and 26°5 respectively. INFLUENZA The followimg example (Table III) is taken from Nothnagel s Encyclopedia of Practical Medicine, English Edition. Article ‘Influenza,’ p. 571. The actual figures for the case incidence and the population imcidence in the various groups are not given, but only the percentage incidences in each case, in the form of graphs. The figures are only approximately correct, as it was not possible to calculate them exactly from the graphs. As in Table II, by dividing the percentages in column 4 by the corresponding ones-1n column 2, we get a series of figures (column 6) which represent. the ratios which the incidence rates bear to one another, and in column 7 the ratios of these rates per cent. Thus, the ‘lability to attack’ (column 7) in the age period 21-30 is slightly more than twice as great as in the age-period 51-60, but what the actual figures for liability to attack are it is impossible to say, as it is only the percentage and not the actual number of people in the groups that is known. The figures in column 7 are not comparable with those in column 4; strictly speaking, no conclusions as to ‘liability to attack’ can be based on the figures in column 4 by themselves. It is only if we assume some knowledge of the number of people in the groups that the case incidence figures have any value in this respect. *The ratios, but of course not the same actual figures, in this column could equally well be got by dividing the figures in column 3 by those in column 2. 203 Thus, we could probably infer that the liability to attack was greater in the 21-30 period than in the 11-20 period, because we assume that the population of the 21-30 period is probably not twice that of the Taste III. Showing incidences and ratios of incidence rates in Influenza. eee is a Incidence Total | Number | Incidence,, Number | rate, i.e., | Ratios of number | of people | i.e., total | of cases | number | Ratiosof the of people that occur} number | that occur| of cases the | incidence that occur, in each of cases ineach | occurring | incidence rates to Age-Group in each group observed | group among ratesto | one group | per 1oo | that occur| per 100 | 100 people one | another (Census) | people in each cases in each another | per cent. (Census) group group aay arr | I-Io0 19 8 o42t | 5°86 | II-20 16 15 0°937 | 13°05 21-30 21 32 1°523 | 21°07 | 31-40 | 16 20 1°250 17°40 41-50 | 12 | 14 | 1166 4 6| 16°23 | 51-60 8 | 6 0°750 10"44 61-70 | 5 | 4 | o*800 | Il'I4 | | 71-80 | 3 | Buncw 0°333 | 4°63 | | | 100 | 100 7180 | 99°82 | | | 11-20 period; but we can only make accurate deductions, giving relative or actual figures, when we base them on the number of people, relative or actual, in the groups. BLACKWATER FEVER It has been commonly stated that the liability to an attack of blackwater fever is greater in persons infected with malignant tertian parasites than in those infected with simple tertian or quartan parasites. These statements are based on the particular parasites present in so many cases of blackwater fever, but, as we have shown 204 above, no conclusions can be drawn as to liability to attack unless we have population data as well. The case before us is parallel with the two examples we have already considered, though here, instead of age-groups, we have groups of persons (malaria cases) infected with the malignant tertian and simple tertian parasites respectively (Table IV). Taste IV. The data are Showing relative liability to an attack of Blackwater fever of persons infected with malignant tertian and simple tertian parasites respectively. I 2 3 4 5 6 7 Incidence Number Total Number | rate, 1.e., Total of cases of |} number | of cases of | number | number | Malaria of Blackwater of cases of | Ratios of | of case of | that occur | Blackwater| that occur Blackwater| incidence | Ratios of Parasite Malaria in each cases ineach that occur| rates to | incidence Group that occur) group observed group among one rates in each per 100 |that occur} per roo too cases | another | per cent. group cases of in each cases of of Malaria Malaria group | Blackwater in each group Ex. 1 | Malignant tertian 74 764 1°032 53°2 Simple | tertian 26 } 23°6 | o"908 46°38 100 | 100 1°94 100° Ex. 2 | Malignant tertian 68°45 54°03 o'789 351 Simple tertian 31°56 | 45°96 1°456 64°9 | | | soe 99°99 2°245 99°9 taken from a paper in the Aznals Parasitology, Vol. VII, December, of Tropical Medicine and 1913, p. 487, in which I have summarised the data of Deeks and James, and Lovelace, respectively . 205 As before, the figures in column 6 are got by dividing those in column 4 by the corresponding ones in column 2. The figures in column 7 are then calculated for cne hundred cases. In the first example, the incidence rate of blackwater fever in malignant tertian infections is only slightly greater than that in simple tertian infections. In the second example, the incidence rate in simple tertian infections is nearly twice as great as that in malignant tertian infections. We are not concerned here with the discrepancy between the results, but with the fact that in each case deductions based solely on the incidence, 7.e., occurrence of malignant tertian or simple tertian parasites in the blackwater cases, would have led to different but erroneous conclusions. A reference to the current text-books of Tropical Medicine would afford many other examples of a similar kind, where conclusions are drawn from a knowledge of the number of cases only, in the absence of any knowledge of the number of people among whom the cases occur. 207 THE EXPERIMENTAL INFESTATION OF PHYSOPSIS AFRICANA BY F. G. CAWSTON, M.D., Cantab. First Streatfeild Research Scholar (Received for publication 25 May, 1922) Some notes on the experimental infestation of Physopsis africana in Natal may prove of use for reference to workers in other parts of the world who are engaged in the study of the life-history of the schistosomes. It was some time before I succeeded in keeping this common fresh-water snail alive for any length of time under artificial conditions. The glass jars in which I observed the growth of young examples proved unsuitable for more than a few days; but wooden tubs, kept out of doors in a shady place, answered the purpose very well, and I was able to secure all I needed whilst the experiments were in progress. I did not find it necessary to change the water in the tubs, which contained a few water-plants and an increasing amount of decomposing leaves and small pieces of wood which fell in occasionally. The snails had, therefore, plenty of shade, whilst the water never became too hot, as it tends to do in a glass jar if placed in the sun. As ‘millions’ had been observed feeding on young snails, and as one did not wish to interfere in any way with the free ventilation of the water, nothing was done to prevent the breeding of mosquitoes. The surface area of the water was approxi- mately double that of the bottom of fhe tubs. It is possible that the rate of growth was handicapped by the food supply—and I have not succeeded in getting the common variety of water-lily to thrive in wooden tubs—but, even under what appear to be very favourable conditions, I do not find it possible to obtain examples large enough for experimental purposes under five months in Natal, and I should gather that by far the majority of 7 Fic, 1. Ova from urine of Natal boy. Note solid, long, rounded extremities. Average length 0:22 mm.; exceptional length 0.2625 mm.; abnormally bent end in I. Living miracidia in 1,2 and 5. Degenerative effects of emetine in 3, 4 and 6. 209 infested examples that I have found in the rivers and pools of South Africa were at least a year old. I have noted the presence of apparently mature cercariae in very small specimens from Natal rivers, and in some which were experimentally infested forty-six days before and the shells of which measured only 6°5 and 7:0 mm. in length; but it is rare to find such small specimens infested. When required for the experiments, a number of well-developed examples, about 12 mm. in length, were selected and placed in a glass jar containing fresh water. The urine of a Bilharzia patient was then secured, and the ova collected by centrifugalising the specimen. The ova were examined microscopically, identified by Fic 2. Ova of S. haematobium from same urine, showing (B) living miracidium about to hatch, and (4) miracidium degenerated under the influence of emetine. means of their shape, size and spine as those of Schistosoma haematobium, and, as soon as the contained miracidia were seen to be ready to hatch, were emptied into the jar containing the snails and placed in a good light for a few hours. At the end of twenty- four hours the snails were then generally placed in a small wooden tub. Some of the snails which had been thus exposed to infestation were placed in some dark glass jars containing a few decomposing leaves. The water in these jars were continually replaced by drops 210 from a glass tubing connected with a large tub containing water- weeds. Whilst the snails were thus continually receiving fresh water laden with food, the water was gradually escaping through a regulated syphon tube. This arrangement answered well for a few snails at a time. Long spindle-shaped ova resembling those of Schzstosomum bovis were found in the urine of two Natal-born Indian boys, associated with the typical ova of Schistosoma haematobium. Both varieties were added tc water containing Physopsis which had been kept free from all other possible chances of infection by muiracidia. O < Cmmia ture ) Fic. 3. Cercariae present in Physopsis africana 63 days after being exposed to the above ova. (X2.) To imitate natural conditions, as far as possible, it is necessary to wait until cercariae are escaping into the surrounding water before using them for the experimental infection of animals. Although I have found apparently mature cercariae in Physopsis which has been exposed to miracidia only a fortnight before, I have never found the cercaria outside a snail until the development has been allowed to progress for thirty-five days, and it is probably better to keep the infested snails living for several months before dissecting them. To ascertain whether the experimental infestation has been 211 successful in specimens one does not wish to destroy, it is best, as Dr. J. G. Becker once pointed out to me, to place individual snails in clean test-tubes in a good light, or even break off a minute portion of the shell over the liver. There may be certain conditions in the surrounding medium that encourage the mature cercariae to work their way out of the infested snail; but I have carefully examined specimens for several days, up to the sixty-fourth day, without any sign of free-swimming cercariae, when dissection revealed the presence of a number of mature cercariae within the liver substance. Among about thirty individuals that I have found infested with schistosomes within one or two months after being exposed to the ova of Schistosoma haematobium and those resembling S. bovis, I have never seen any cercaria which shows eye-spots, development in rediae or possessing the long prongs that some of the schistosomes that I have found in Physogsis in the Natal rivers occasionally do. In every instance, when mature, the experimentally produced cercaria in Physopsis exposed to infection from the urine of a Bilharzia patient was 0525 mm. in total length, possessed prongs which were about a quarter the length of the tail, and in other respects resembled the cercaria of S. haematobium. REFERENCES Cawston, F. G. (1921). Experimental Infestation of Fresh-water Snails. Trans. Royal Society of South Africa, Vol. IX, Pt. 4, p. 301. 7 (1921). Three Schistosomes in Natal which possibly attack Man. ‘Yourn. Trop. Med. and Hyg., Vol. XXIV, No. 18, pp. 242-244. . 213 NOTES ON CULICIDAE IN VENEZUELA, WITH DESCRIPTIONS OF NEW SPECIES PART II BY ALWEN M. EVANS (Received for publication 31 May, 1922) PLATE XI Since the completion of a previous paper (1921) we have received, through the kindness of Dr. Chacin and Dr. M. Nufiez Tovar, a number of consignments of mosquitoes collected, during the Autumn of 1921, from the regions surrounding Caracas and Maracay. Most of the species represented are common, and of wide distribution, but among them are a new species of the Avribalzagia group of Anopheles and a very distinctive new species of the Janthinosoma group of Psorophora. Anopheles albimanus, Wied. La Cabrero, Estado Carabobo, 21; Tapatapa, near Maracay, ¢ I, 293; La Barraca, Maracay, 993. Dr. M. Nunez Tovar. A. albimanus var. tarsimaculata, Goeldi. San Francisco, near Maracay, 9? 3 ; La Cabrero, Estado Carabobo, 924; La Barraca, near Maracay, ?1; near Maracay, 332, 21. Dr. M. N. Tovar. A number of specimens occurred in which the condition of the palpi was intermediate between the type A. albimanus and the variety tarsimaculata. Some specimens have the palpi with the penultimate joint with basal white ring ; beyond, black scaled with a number of white scales scattered among the black ones about half-way between base and apex of segment. In other cases the-penultimate joint with basal white ring and most of scales on outer side of a much paler brown than the rest of the dark palpal scales. 214 A. argyrotarsis, R.D. San Francisco, near Maracay, 21. Dr. M. N. Tovar. A. pseudopunctipennis, Theo. San Francisco, near Maracay, 33 3, 22 2; La Cabrero, Estado Carabobo, 9 1; Tapatapa, near Maracay, ¢ 1, 9? 3; La Barraca, near Maracay, 9? 2. Dr. M. N. Tovar. A. maculipes, Theo. San Francisco, near Maracay, 21. Dr. M. N. Tovar. Anopheles (Arribalzagia) venezuelae, sp.n. (Plate XI). FEMALE. Proboscis slightly more than 3 mm. Curved ventrally in distal half ; labellae conical, dark ochraceous distally, shading to brown at base with a few fine dark hairs; vestiture of dark coppery brown scales roughened basally beneath. Palpi shorter than proboscis, clothed with black, spatulate scales sub-erect on basal third, yellow scales forming very narrow rings at apex, at base of last segment, and at middle of long segment ; also a few scattered pale scales on distal half. Apex with a tuft of pale yellow hairs. Antennae with long segments densely clothed with fine decumbent hairs, setae of whorls sparse, pale. Third segment with a large patch of flat decumbent scales on upper and inner side on distal half, white proximally, ochraceous golden distally. Tori moderate, dark brown with white scales externally. Clypeus large, surface minutely punctate, olive brown proximally, ochraceous pollinose distally. Eyes deep black. Occiput with median grove ; integument brown, light grey at margins of eyes and medially. Vestiture of dense upright forked scales, ochraceous in front, with a few whitish ones in middle, dark ones behind; posteriorly and laterally scales very dense, black. Space between eyes with pale narrow scales at borders, and many long creamy forwardly-projecting hairs. A few brown setae projecting forwards from posterior margins of eyes. Prothoracic lobes with erect black scales and a few brownish hairs above ; whitish scales below. Mesonotum greyish pruinose mottled with brown spots ; three large black ocellar spots : two lateral and one posterior involving the scutellum. Vestiture of straw-coloured hairs, many of them arising from pigmented spots. Median area bordered by two 215 narrow longitudinal yellowish depressed bare stripes, and two wider bare depressions extending from the lateral ocellar spots to the posterior border of the mesonotum. At anterior border a group of strongly curled, pale yellow hairs. Scwutellum greyish pruinose at sides, along posterior border a median and two lateral groups of yellow hairs and a continuous row of long dark setae. Postnotwm nude, dark tawny ochraceous at sides with broad brown median stripe. Pleurae pale greyish pruinose with five large spots unicolorous with ocellar spots on thorax, dark brown basally. A few flat, white scales medially. Abdomen dorsum uniform mouse grey, vestiture of fine yellow hairs. Distal margins of segments two to seven with a few black scales, and lateral projecting tufts of black spatulate scales with a few white ones near them. Eighth segment with pre-apical band of flat ochraceous scales, and border of flat black scales, and pale yellow hairs. Last segment black scaled. Venter greyish pruinose, with long yellowish hairs at sides of segments, and shorter dark hairs medially. Segments two to seven with broad, white semi-decumbent scales thinly scattered over surface ; apical third with dense patches of sub-erect black scales. Venter of eighth segment with golden-yellow, appressed scales. Terminal segment black scaled. Wings. First fork cell one and a half times as long as its petiole ; second fork cell about as long as its petiole. Basal cross vein very narrowly separated from anterior cross vein. Three large black scaled patches bordered with white scales, with membrane beneath deeply infuscated, the largest median involving the costa, sub-costa and first and second veins ; the smallest basal, involving the costa, sub-costa and first vein ; the distal one involving the costa, first vein, and both branches of the second vein. Pale scales of wings mostly light yellow except white spots bordering large black patches. Costa mostly black scaled with ten small pale spots, four proximal to large median black patch ; scales at apex of costa grey. Sub-costa mostly black scaled with small pale spots opposite those on costa, some decumbent pale scales on basal quarter, largely pale scaled beyond median black patch, apex pale. First vein, in addition to the three large black patches, with black decumbent scales on proximal eighth, between proximal and distal black patch, outstanding scales mostly pale, decumbent scales black ; between median and distal black patches, vein mostly pale scaled, with one small black spot and a few scattered black scales. Beyond distal dark patch two grey spots ; 216 apex of vein pale. Second vein: stem beyond median black patch mostly pale scaled, upper branch of fork black scaled on proximal half, beyond with pale scales and a few grey ones; apex grey. Lower branch of fork with a large proximal and apical dark patch, and two smaller dark spots separated by a pale patch. Third vein dorsally mostly pale scaled, with four black spots, two near the base, one apical, and one sub-apical ; one or two black decumbent scales on central, pale area. Third vein from below appearing dark scaled. Fourth vein mostly black scaled, stem with many pale decumbent scales between the black ones. Proximally, rest of stem with seven pale spots, one at base of fork ; branches of fork each with three pale spots; on upper branch the middle white spot elongate. Fifth vein mostly pale scaled, stem with three black spots at base, and many decumbent black scales on distal half. Upper branch of fork with four small and one large black spot, apex black; lower branch with two black spots distally, apex pale, both branches with a few black scales scattered among the pale ones. Sixth vein with seven black spots ; apex black. Fringe with twelve pale spots, largest at apex of lower branch of fifth vein. Outstanding scales varying greatly in size and shape ; on distal portions of veins lanceolate to narrowly ovate ; on basal portions ovate with many very broadly ovate ones on fourth vein and sub-costa, some of those on sub-costa obliquely truncate. (Plate XI, fig. I). Halteres above densely white scaled with sub-circular dark median area ; beneath densely black scaled, stem nude, ochraceous. Legs. Very long and slender. Vestiture black with many white spots and bands. Femora and tibiae densely mottled with white spots. Hind tarsi with ten white rings or spots on first joint, seven on second, four on third ; fourth and fifth white ringed at base and apex and in middle (Plate XI, fig. 2). Front tarsi with eleven white rings or spots on first joint, five on second, four on third and two on fourth and fifth. Third tarsi with ten white spots on first joint, five on second, four on third, three on fourth ; fifth white with two narrow black rings. Length 6-5 mm. Wing 5°5 mm. One @ taken at’La Cabrera, Estado Carabobo, Autumn, 1921, by Dr. M. Nunez Tovar. This large and beautiful species approaches closely in the markings of the wing to Anopheles (Arribalzagia) punctimacula, Dyar and Knab, as described by Howard, Dyar and Knab (1917), under the name A. malefactor. The chief differences are tabulated below. 217 A. punctimacula, D. and K. A. venexuelae, sp.n. Length ax Ss os ... About 5:omm. ... ae -- 65 mm. Wing . About 475mm. ... oe «| 5°5 mm. Vein III vide ee ae --- ‘Two small spots at and near _ Base pale, two small black spots base and two others at and near base, two others at and near apex, a few black scales near apex, only one group of scattered along its whole length.’ | two small black scales on rest | of upper surface of vein. Distance between anterior and About equal to length of basal | Less than a quarter of the length basal cross veins cross vein | of the basal cross vein Hind tarsal segments 3 ax .-- Lapical, 1 basal, and 1 median, 2 white rings between white white ring apical and basal rings Hind tarsal segments 4 <5 ... As third segment Sas ... With 1 apical, 1 basal, and I median ring Hind tarsal segments 5 20 ... Entirely white (or with @ black With two black bands band)* *Dyar, 1918 Limatus durhamii, Theo. La Barraca, near Maracay, @ I. Culex quinquefasciatus, Say. Houses and buildings in and around Caracas, 33 275, 22 738. Dr. Chacin. About 100 3¢ were determined by the genitalia. Near Maracay, 33 8, 99 4. Dr. M. N. Tovar. Culex declaratory, D. and K. : Near Maracay, 1, 91. Dr. M. N. Tovar. Culex corniger, Theo. Caracas, d I, 21. Dr. Chacin. Aédomyia squamipennis, Theo. Near Maracay, 99 2. Dr. M. N. Tovar. Mansonia titillans, Walker. Near Maracay, 99 2. Dr. M. N. Tovar. Psorophora posticatus posticatus, (Wied.) Dyar. Near Maracay, dd 2, 995. Dr. M. N. Tovar. 218 Psorophora posticatus sayi, D. and K. Near Maracay, 91. Dr. M. N. Tovar. Psorophora lutzii, Theo. Near Maracay, 99 2. Dr. M. N. Tovar. Psorophora saeva, D. and K. Near Maracay, 99 2. Dr. M. N. Tovar. The specimens differed from the description of P. saeva in the mono- graph of Howard, Dyar and Knab (1917), in having the scales of the proboscis sub-erect. Psorophora ciliata (Fab.) R.D. Near Maracay, 9? 2. Dr. M. N. Tovar. Psorophora (Janthinosoma) tovari, sp.n. FEMALE. Pyroboscis uniform; labellae small, conical; vestiture of dark scales with violet reflections. Palpi about one-fifth of the length of the proboscis, curved, with partially erected black scales with violet reflections, and a few rather long, coarse setae. Antennae: long segments very dark brown, with delicate white decumbent hairs, setae of whorls blackish brown. Tori ochraceous externally, dark brown internally, dark area with a row of pale flat scales and a number of dark setae. Clypeus large, very shining black above, ochraceous at sides above, the colours separated in a distinct line at sides below shading to dark brown. Eyes large. Occiput wide; integument dark shining blackish-brown above, tawny below. Vestiture of broad very much curved (much more strongly curved than the pale scales on the mesonotum of P. posticatus, Wied.) creamy white scales scattered over entire surface, intermixed with creamy upright forked scales on median third, and with broad curved yellowish scales and dark brown setae on lateral thirds; coarse dark setae anteriorly, and a tuft of ochraceous setae projecting between eyes. Posteriorly, upright forked scales black. Prothoracic lobes with silvery, much curved scales, and numerous very coarse, black setae. Mesonotwm: integument very dark brown, dull. Vestiture on centre of disc of narrow curved, brown scales with brassy reflections, these scales extending laterally behind almost to wing roots. 219 Sides of disc with broad much curved, creamy scales resembling those on occiput intermixed with a smaller number of bronzy, broad, curved scales. A median band, narrowed behind, of broad, much curved, creamy scales, on anterior sixth. Posteriorly mesonotum partially denuded, antescutellar space dotted with spindle-shaped ochraceous and creamy white scales intermixed. Plewrae: integument dark, shining, sepia ; above with creamy scales as on sides of mesonotum ; below with numerous broad, flat white scales. Scwutellwm with broad, flat, creamy and yellowish white scales. Postnotum nude, dark brown, shining. Abdomen. First segment with a median broad patch of creamy white, flat scales, scattered pale ones at sides and numerous fine pale hairs ; laterally a conspicuous patch of creamy scales not visible from above. Segments two to six with dark bronzy scales with brilliant metallic blue reflections, and conspicuous apical bands of creamy yellow scales, continuous at sides with pale scales of venter. Segments two and three with creamy band narrowed at sides, and produced backwards medially forming a wide triangle with apex approaching within a third of the base of the segment. On segment four, median backward extension of band broader, truncated. Segments five and six with band broadest in middle, gradually narrowed at sides. Segment seven mostly pale scaled above. Venter entirely clothed with pale golden and silvery scales, but proximal half of second segment denuded. Wings : Membrane deeply infuscated. First fork cell one and a half times the length of its petiole ; second fork cell slightly longer than its petiole. Basal cross vein separated from anterior cross vein by nearly its own length. Outstanding scales sepia, ligulate. Halieres: knobs brown, stems ochraceous. Metatarsi of hind legs with no sub-erect scales. Front and mid-femora pale straw coloured with dorsal broad stripe and apical narrow ring of bronzy scales with metallic violet reflections. Hind femur with bronzy metallic scales. Tibiae and tarsi clothed with dark bronzy metallic scales, a stripe of scales with ochraceous reflections on under sides of front and middle tibiae, lines of scales with brassy reflections on under sides of the metatarsi and tarsi. Hind legs with segments three, four and five of tarsi missing. Claws of first and second tarsi with teeth :—1-I—1r-1. Length c.5 mm. Wing c. 4 mm. Two 992 taken in region of Maracay, Venezuela. Dr. M. Nujez Tovar, 1921. Aédes argenteus, Poiret. Houses and buildings in and around Caracas, 33 28, 92 327. Dr. Chacin. Near Maracay, 33 18, 996. Dr. M. N. Tovar. Aédes trivittatus (Coq.), D. and K. Near Maracay, 91. Dr. M. N. Tovar. Aédes (Finlaya) oswaldi, Lutz. Near Maracay, 91. Dr. M. N. Tovar. REFERENCES Dyar (1918). Ins. Ins. Mens. Vol. VI, p. 147. Evans (1921). Ann. Trop. Med. & Parasit., Vol. XV, p. 445. Howarp, Dyar, and Knas(1917). The Mosquitoes of North and Central America and the West Indies Vol. III. While the present paper was in the press an extensive collection of Arvibalzagia sp. from the Panama Canal Zone has been obtained. A preliminary examination of these specimens leads me to consider that the characters on which Dyar (1918) defines the species of Arvribalzagia in his tables may be extremely variable. It seems probable that, although the type of A. venezuelae does not fall under any of the species in this table, and does not agree in detail with any of the descriptions of the existing species, it is in reality a variety of A. punctimacula, D. and K. A detailed examination of the material is being made, and a further note on the subject will be published shortly. 222 EXPLANATION OF PLATE XI Anopheles (Arribalzagia) venezuelae, sp.n. Fig. 1. Wing. Fig. 2. Last three segments of hind tarsus. Both figures drawn with camera lucida. Annals Trop. Med. & Parasitol., Vol. XVI PLATE X1 223 ANCYLOSTOMES RECORDED FROM SIXTY-SEVEN POST-MORTEMS PERFORMED IN AMAZONAS BY R. M. GORDON From the Research Laboratory of the Liverpool School of Tropical Medicine, Mandos, Brazil (Recezved for publication 12 June, 1922) This paper deals with the ancylostomes collected at sixty-seven autopsies, performed in the Santa Casa Hospital, Mandos, during 1921 and the beginning of 1922. With very few exceptions, all the subjects had resided for the greater part of their lives in the State of Amazonas, Brazil. They divided themselves into two natural classes :—(1) The ‘ Town-dwellers’ and (2) the ‘ Country-dwellers,’ the latter mostly agriculturalists, rubber-workers, etc., who either came into the town to be treated for sickness, or else who were taken ill when temporarily residing there. METHOD OF COLLECTION. The gut having been opened, all ancylostomes, attached or lying loose in the lumen, were removed. The contents of the bowel were then distributed in large, flat, white dishes and examined for ancylostomes during three washings. All worms were washed in normal saline, killed with hot 75 per cent. alcohol, and stored in lacto-phenol (Leiper). METHOD OF EXAMINATION. In the first part of the investigation an attempt was made to estimate the accuracy of a hand-lens (x 8) examination of the worms in order to determine sex and species. For this purpose, the worms obtained from fifteen post-mortems were examined as follows :—First with a hand-lens and a tentative diagnosis made as to sex and species (Z.e., whether Necator americanus or A. duodenale). They were then re-examined with a microscope, using the half-inch and the one-sixth. The points of distinction noted, during the hand-lens examina- 224 tion, were (1) the general fineness, and (2) the sharply defined head curve, of Necalor americanus as compared with A, duodenale. In this manner six hundred and sixty-two worms were examined ; these consisted of eighty-five A. dwodenale and five hundred and seventy- seven Necator americanus. The result was as follows: —One worm was diagnosed wrongly (Necator americanus male, mistaken for A. duodenale male); three other worms necessitated microscopical examination, but two of these proved to be so damaged that the head curve was destroyed ; the remaining six hundred and fifty-eight worms were found to have been correctly diagnosed with the hand- lens. With a view to testing whether A. caninum or A. braziliense could be distinguished from A. dwodenale and A. necator, one male and one female A. caninum and one male A. braziliense (all from a cat) were mixed with sixty Necator americanus and eighteen A. duodenale. The worms were then separated into their species by the aid of a hand-lens, the result checked by a microscope and found to be correct. The distinctions between A. xecator and A. caninum or A. braziliense were based on the characteristic head curve of A. necator, between A. duodenale and A. caninum or A. braziliense on the smaller size and general fineness of the latter two species. As this method appeared sufficiently accurate, the worms were sorted with a hand-lens in all subsequent examinations, any doubtful specimens, and these averaged one in sixty, being placed on one side and subsequently examined microscopically. RESULTS. These are published in the form of a table for comparison with Darling and Smillie’s (1921) figures for Brazil. Apparently their results are drawn from Southern Brazil, chiefly from Rio, Pernambuco, Sao Paulo, and a few from the State of Matto Grosso. They state that ‘the groups studied were all more or less similar in that they were composed largely of agriculturists. The average hookworm count of 136°1 per case, therefore, does not represent the degree of infection of all Brazil, but of rural Brazil.’ As my results are drawn from two classes, a second table is published showing a comparison between town and country infec- tions. It must be noted that a few of the cases recorded had at one 225 time or another been in hospital, and a certain number of those had undoubtedly received Chenopodium. On examining Table I, it will be seen that the most striking difference between the figures for Amazonas and South Brazil lies in the proportion of Necator americanus to A. duodenale, and, on examining Table II, that this difference is mainly due to the high average number of A. duodenale occurring in the country dwellers. Whereas Darling’s rural dwellers for South Brazil show a proportion of Necator americanus to A. duodenale of 45 to 1, rural dwellers in Amazonas show a proportion of only 3°2 to I. Ancylostoma braziliense in human beings. Four worms belonging to the species A. draziliense were found among the six thousand eight hundred and fifty-seven ancylostomes, collected from the sixty-seven post-mortems. There were two males and two females; the males measured about 7 mm. in length and the females 7°5 mm. Each worm was found in a separate host. Two were found in native Amazonians who, so far as is known, had never left the State of Amazonas; one in a patient who originally came from Ceara, and one in an American of the ‘beach comber’ type who had lived some twenty years in North Brazil. I can find no previous record of A. brasiliense being found as a human parasite in America. De Faria (1916) states that he examined children in Rio for this infection without success. Darling and Smillie (1921) do not record it among the sixty three thousand nine hundred and twenty-three hookworms they examined in South Brazil; but Darling (1920) writes :—‘ The ancylostomes encountered in man are A. duodenale, A. ceylanicum, A. braziliense, Necator americanus. 1 cannot, however, find the authority on which A. braziliense is included. According to de Faria (1910 and 1916) and Clayton Lane (1916), the distinction between A. braziliense and A. ceylanicum depends on the following two points :— (1) The inner ventral tooth. This is smaller and finer in A. braziliense than in A. ceylanicum. (2) The bursa of the male. De Faria (1916) states that in A. braziliense the rays, especially the dorso-external, are characterised by their great length, fineness and delicacy, whilst those of A. ceylanicum are shorter and thicker. 226 Taste I Comparing Ancylostome Infections for Amazonas and South Brazil. Amazonas July, 1921 to February, 1922 South Brazil (Darling) April, 1918 to January, 1920 Number of cases examined 67 469 Number of Ancylostomes found 6,857 63,923 Number of Necator americanus 5,660 62,554 Number of 4. duodenale 1,193 1,369 Number of 4. brazxiliense 4 — Proportion of Necator americanus to A. duodenale ... 47:1 45:1 Average number of Ancylostomes to each individual 10273 136°1 Average number of Necator americanus to each individual 8474 133°2 Average number of 4. duodenale to each individual 178 2°9 Taste IT. Comparing Ancylostome Infection of Country and Town Dwellers in Amazonas. Country Town Dwellers Dwellers Number of cases examined 39 28 Number of Ancylostomes examined 4,144 2,713 Number of Necator americanus 3,157 2503 Number of A. duodenale 985 208 Number of A. braziliense 2 2 Proportion of Necator americanus to A. duodenale ... Bee 12:1 Average number of Ancylostomes to each individual 106°2 96°8 Average number of Necator americanus to each individual 80°9 89°3 Average number of A. duodenale to each individual 25°2 74 227 The distinction between the two was disputed by Leiper (1913). I have had the opportunity of comparing the following ancylostomes :— (1) A. braziliense from cats and dogs in N. Brazil. (2) A. braziliense from human subjects in N. Brazil. (3) A. ceylanicum from cats and dogs in Bengal, India. (Material kindly supplied by Lt.-Col. Clayton Lane.) (4) A. ceylanicum from West African dogs and South African cats. As a result of careful examination of many specimens, I was unable to confirm the specific differences mentioned by de Faria and Clayton Lane. No constant difference could be detected in the size and shape of the inner tooth of A. dvazilense and A. ceylanicum, nor could any difference be discovered in the length and fineness of the dorso- external ray in the two worms (v7de table). Taste III, Comparing Measurements of the Dorso-external Ray in A. ceylanicum and A. braziliense 5 | Ratio | Ratio Average length | breadth | Number | length | Average | Average | D.E.R. | D.E.R. As named Locality | Host | examined of breadth | length to to worm | D.E.R. | D.E.R. | length | length worm | worm mm. 7 Le A. ceylanicum | Berhampore, Cat 3 So. 14 «|- 176 1:31 | 13400 Bengal A. ceylanicum | Berhampore, Dog ... I 75 21 217 Teme Melgs7 Bengal A. ceylanicum | Accra, Dog ... 3 70 20 270 1:25 | 1:350 West Africa A. braziliense | Manaos, DOR, po. 4 6°5 17 17I 1:38 | 1:382 North Brazil A. braziliense | Manaos, | Cate ea. 3 61 17 186 Yisg2) (wsges North Brazil A. braziliense | Manaos, Human 2 70 14 162 1:43 | 1: 500 North Brazil 228 SUMMARY Six thousand eight hundred and fifty-seven ancylostomes collected from sixty-seven autopsies performed in Manaos, Amazonas, were examined, with the results recorded. A far higher proportion of A. duodenale to Necator americanus (1 : 4°7) occurred in Amazonas than recorded by Darling for South Brazil (1 : 45). This high proportion of A. duodenale was shown to be chiefly due to the country dwellers in Amazonas, whose A. duodenale to Necator americanus ratio was 1 : 32, while that of the city dweller was it 9 1D A. braziliense was found in four of the post-mortems. The comparison of these worms and other two-toothed ancylo- stomes from dogs and cats in N. Brazil and India, and also from cats in South Africa and dogs in West Africa, failed to show the difference claimed to exist by de Faria between A. ceylanicum and A. braziliense. My thanks are due to Dr. Thomas for much of the post-mortem material. REFERENCES Daruine, S. T. (1920). Observations on the Geographical and Ethnological Distribution of Hookworms. Parasitology, Vol. XII, No. 3, p. 217. Darwin, S. T., and Smitti, W. G. (1921). Studies on Hookworm Infection in Brazil (First Paper). Monographs of the Rockefeller Institute for Medical Research, No. 14. Gomes pe Farra (1910). Contribution towards the Classification of Brazilian Entozoa. Mem. Inst. Os. Cruz, Vol. II, No. 2, p. 286. ——— (1916). Nota Sobre Agchylostoma braxiliense (G. de Faria, 1910). Mem. Inst. Os. Cruz, Vol. VIII, No. 2, p. 71. Lanz, Crayton (1916). The Genus Anchylostoma in India and Ceylon. Ind. Fourn. Med. Res., Vol. IV, No. 1, p. 87- Lerrer, R. T. (1913). The apparent identity of Agchylostoma ceylanicum (Looss, 1911) and Agchylostoma biaxiliense (Gomes de Faria, 1910). ‘Journ. Trop. Med. and Hyg., Vol. XVI, p. 334- 229 (PPE *-SUSGEPTIBILIPY: “OP” “THE INDIVIDUAL _.LO,...c HE. BrrEs OF STEGOMYIA CALOPUS BY R. M. GORDON From the Research Laboratory, Liverpool School of Tropical Medicine, Manaos, Brazil (Received for publication 27 May, 1922) The usual belief amongst Europeans residing in the Tropics, with regard to the susceptibility of the individual to the biting of mosquitoes, would appear to be that the new-comer receives propor- tionately more bites than the old resident, but that the native of the country receives less than either. Marchoux, Salimbeni and Simond (1903), writing of S/egomya calopus, state ‘ . .. Il a une prédilection marquée pour la race blanche.’ And later in the same article, ‘Il s’attaque beaucoup plus avidement aux individus jeunes, vigoureux, qui ont la peau fine et le teint coloré, qu’aux individus anémiés ou agés.’ It appeared of interest to test the truth of this idea and, at the same time, to investigate the following points with regard to their influence on the biting of mosquitoes: (1) Sweating ; (2) hairiness of skin exposed to bites; (3) colouration; (4) age. Attention was also paid to the subsequent local reaction to the bites. Nature of Experiment. All experiments were performed with Stegomyia calopus, owing to its being a day-feeder and the commonest mosquito in the locality. Sixteen experiments were performed, at each of which a number of male persons, usually six, of various nationalities and different lengths of residence in Brazil, were exposed, under the same conditions, to the bites of a number (usually forty-five to fifty) of 230 hungry Stegomyza calopus females. The number of completed feeds performed on each individual during 30 minutes were noted. Apparatus used and Method of Recording Results. The feeding- box consisted of a large mosquito cage measuring 24 by 15 by 12} inches, and fitted with six sleeves. All experiments were performed in daylight at approximately the same hour, and the box placed in such a position that it was as far as possible evenly illuminated. Mosquito cage used for the experiment. In practice, the sleeve was fitted closely to the forearm, at its junction with the cage wall. Forty to fifty female mosquitoes, which had been kept unfed for at least four days since their date of emergence, were released in the cage. The individuals to be tested then introduced one of their hands through the sleeves so that each had the same amount of forearm and hand exposed to bites. It was found in practice that the female S/egomyza, unless disturbed, never bit twice, and counts could be made easily and accurately. Name L.O. After the first two experiments it was noted that mosquitoes that bit on the under surface of the wrist and hand were hard to count, and, at Dr. H. Wolferstan Thomas's suggestion, in all subsequent 231 experiments, cardboard shields were used to protect this surface. An example of an experiment is given to show the data recorded. Sweating: Sweating : | EXPERIMENT 3. Date: 3.10.21. Number: Stegomyia calopus = 50. | | | | | | Residence | Hairiness | Number in other Age Colouration | of exposed | Nationality | of years countries | first arm in Brazil | with service | 15 mins. : . il . h . 5 . : in each 30 | Dark ° British 9 England + / only mee) 96 gold Dark ° Portuguese | 16 Portugal + 20 years Pleas Fair ° British 2 England ° ; and Canada only 20 Dark ° Brazilian 18 Nil re) 47 | Fair + Canadian 15k England re) | | and | | | Canada | only eles Dark + | Brazilian 23 Nil - ; REsuLts. I. Length of Residence. Total Total number of | number of | | individuals — bites tested received Persons above 5 years’ but under 30 years’ residence in Brazil. (Majority 10-15 years) A =| 62 462 Persons under 2 years’ residence in Brazil. (Majority under 1 year, some a few weeks) . wee see 26 157 second 15 mins. + + Average | number of bites received by each individual 74 Number of bites received 232 II. Sweating. Average Total Total number of | number of | number of bites individuals bites received by tested received each individual Persons sweating on the exposed forearm and hand | 30 212 70 Persons not sweating on the exposed forearm and hand. 58 407 70 Ill. Hairiness. Average Total Total number of number of | number of bites individuals bites received by tested received each individual Persons showing a considerable amount of ‘ hairiness’ on the exposed forearm and hand | 40 286 71 Persons not showing any marked ‘hairiness’ of exposed forearm and hand 48 333 6°9 IV. Colouration. Average Total Total number of number of | number of bites | individuals bites received by tested received each individual Persons of a dark colouration (‘ dark’ being used in the accepted sense of dark eyes and hair) ee 62 441 71 Persons of a fair colouration 26 178 68 V. Age. Average Total Total number of number of | number of bites individuals bites received by tested received each individual Persons of 30 years and under 29 194 6°6 Persons of more than 30, and less than 40 36 277 76 Persons of more than 40 23 148 64 VI. Nationality. Average Total Total number of number of | number of bites individuals bites received by tested received each individual Persons of British, Canadian or American extraction ... 57 352 61 Persons of Portuguese extraction ... 23 193 83 Persons of Brazilian extraction 6 59 98 Persons of Chinese extraction I 7 770° Persons of African native extraction I 8 8:0 Local Reaction. It was found impossible to record this in figures for lack of standard comparisons, but the impression of all observers during the experiment was that, as a class, the new-comers reacted most, the long-resident Europeans less and the native Brazilians least; in the case of the last named, as a rule, no reaction whatsoever could be detected. 234 SUMMARY Before any conclusions can be drawn from these figures, two fallacies must be considered :— (1) To obtain true results each of the sixteen experiments should be considered on its own merits, but this would demand too much space; as, however, it was arranged that as far as possible at each experiment approximately the same proportion of variable factors (z.e., hairiness, nationality, etc.) should be present, and as the proportion of sweating to non-sweating individuals remained nearly constant, it appeared legitimate to add together the number of bites received in the sixteen experiments. (2) The number of individuals tested and the number of bites recorded are so small that no definite conclusions can be drawn; they merely suggest what follows :— Eighty-eight male persons of various nationalities and ages were tested with regard to their susceptibility to the bites of S/egomyia calopus. Six hundred and nineteen bites were received in all. The following factors were recorded :— (1) Length of residence in Brazil; (2) sweating of surfaces exposed to bites; (3) hairiness of skin exposed to bites; (4) coloura- tion; (5) age; (6) nationality. The resulting figures would seem to show that none of these factors exert any marked influence on the number of bites received by the individual. The theory that the number of mosquito bites received by the new-comer is greater than those received by the old resident, both being greater than those received by the native of the country, would appear to be, in part at any rate, attributable to the local reaction immunity displayed by the native, and to a less extent by the old resident. I am indebted to Dr. H. W. Thomas for much help and suggestion. REFERENCE Marcnoux, Sarmsent, and Srmonp (1903). Annales del’ Institut Pasteur. Vol. XVII, p. 694. 235 TUBERCULOSIS IN THE SUDAN, WITH NOTES ON A CASE OF BREAST TUBER- CULOSIS IN A SUDANESE BY BR. 5Ge ARCHIBALD: {D:.S:©:.5M_D. DIRECTOR, WELLCOME TROPICAL RESEARCH LABORATORIES, KHARTOUM (Received for publication 17 June, 1922) PLATE XII It would appear from the number of papers recently published dealing with the subject of ‘ Tuberculosis in the Tropics’ that interest in this disease has been awakened, and in view of its importance, the writer feels little apology is needed for offering some general observations on ‘ Tuberculosis in the Sudan.’ Unfortunately, such observations can in no sense be considered complete, as they are based on a limited amount of clinical and pathological material obtained during the past fourteen years from almost every district of the Sudan. Limited though this material has been, it is in the main representative of a disease which, happily, has not assumed the proportions prevailing in Western countries or even in some of the towns of the Far East. The reason for this is not far to seek. At the present time the Sudan lacks the thickly populated centres of other countries in the West and East. Vast tracts of desert waste and swamp still await to be linked up by means of railways, and although inter- tribal trade and communications have been encouraged under British administration, there still exists in certain parts a conservatism fostered by racial and religious differences, which will take years to break down. Once these obstacles are removed, the seeds of tubercle will assuredly grow and be disseminated on what can only be considered a virgin soil. Any opinion offered as to how and when the disease was first introduced into the Sudan can merely be hypothetical. 236 In the days of Mahdism and up to the time of the British occupation, the country certainly enjoyed a comparative seclusion from the tide of civilization. On the Northern frontier, little inter- communication cccurred with Egypt. The perils attendant on the long desert wastes of this region acted as a deterrent to intimate relations between the two countries, and it is safe to infer that little encouragement was offered to the pagan tribes of the South and West, while on the Eastern Abyssinian frontier, the racial and religious differences of the two countries were sufficient reasons for keeping them aloof. It is, however, to the earlier history of the country that one must turn for information regarding the introduction of tuberculosis; this history, as will be seen from archaeological records, is intimately connected with the ancient history of Egypt. As far back as 2600 B.c. the Northern Sudan was invaded by the Egyptians, and from 2000-1000 B.c. this portion of the Sudan appears to have been occupied by them and regular colonies established as far south as Kerma in the Dongola Province. That tuberculosis was existent among the Egyptians during these periods was established by the late Sir Armand Ruffer (1921), whose work on the ‘ Palaeopathology of Egypt’ is well known. In an admirable ccllection of his studies on the subject, edited by Prof. Moodie, of Hlinois University, there are plates illustrating Pott’s disease in figures discovered in the tombs of Beni Hassan, 2000 B.c. Two other plates also depict graphically Pott’s disease, and a large psoas abscess in a mummy of a priest of Ammen of the XXIst Dynasty, 1000 B.c. Derry’s (1907-08) investigations recorded in the ‘ Archaeological Survey of Nubia,’ apart from representing the first record of tuberculosis in the Sudan, afford circumstantial proof of the intro- duction of the disease from Egypt into Lower Nubia, and it appears reasonable, therefore, to infer that tuberculosis obtained a footing in the Sudan synchronously with the tide of settlers from Egypt, but did not spread throughout the country for reasons already mentioned. At the present time there is little doubt that the disease is practically confined to the larger towns of the Northern Sudan. It is here that the factors concerned with stress, and resultant to a great extent on civilization, play no small part. Overcrowding, 237 intestinal parasitism, malaria, venereal disease, alcoholism and the “hasheesh habit’ undoubtedly predispose to infection by lowering the resistance of individuals peculiarly susceptible to the virus otf tubercle. Amongst the hardy, simple-living nomadic tribes of the desert and the comparatively secluded tribes of the South, the disease is practically unknown; with increasing facilities for inter-communica- tion, however, such a state of things is unhappily not likely to continue. For obvious reasons, more especially when one is dealing with a Mahommedan population, it is impossible to obtain statistics regarding the incidence of, or death rate from, tuberculosis. Racial and religious prejudices often interfere with the calling in of qualified medical aid, and post-mortems are rarely obtained except in cases presenting a medico-legal aspect. Consequently one is compelled to admit that figures obtained from hospitals and dispensaries do not represent the true incidence of tuberculosis in the country, and the writer is of the opinion that such incidence is higher than is suspected. Unfortunately von Pirquet’s test has not been carried out on a sufficiently large scale to permit of any deductions being made. The tribes of the Sudan are very susceptible to such respiratory diseases as bronchitis, broncho-pneumonia and pneumonia, and their predisposition to tuberculosis was referred to many years ago by Balfour (1904). The Sudanese or ‘black’ appears peculiarly susceptible, and it is stated that the Hadendowa, a black tribe inhabiting the hills of the Red Sea, shares this susceptibility. Other observers, notably Bushnell (1920) and Cummins (1920), have called attention to the susceptibility of coloured races to tuberculosis. Moreau (1919) and Roubier (1920) have pointed out the difficulties in detecting the disease among black troops even when the patients are greatly infected, and they prove the value of radiological examination in such cases. The same difficulties are experienced in the Sudan, especially as regards pulmonary tuberculosis, and it may not be amiss to mention here that the disease is at times simulated by bronchial spirochaetosis and a bronchitis of streptococcal origin. The predisposing causes to tuberculosis in the Sudan have already been referred to, and there is no doubt that overcrowding 238 and the filthy habit of expectoration are the determining factors concerned with the spread of the disease, more especially in the cold winter months when overcrowding to the exclusion of light and air favour the possibilities of ‘massed infection.’ Scott’s (192!) observations equally emphasize the rdle played by overcrowding and expectoration as causative factors in tuberculosis among the Chinese in Hong Kong. The view that infected milk is a cause of tuberculosis in the Sudan may readily be dismissed; it 1s true that goats’ milk, cows’ milk, and to a less extent camels’ milk, represent an important feature in the dietary of the natives of the country; nevertheless, tubercular disease of these animals is unknown. Many years ago the writer (1910) recorded a case in which acid-fast bacilli were found in lesions of the lung of a camel simulating miliary tuberculosis, but it should be stated that the possibility of these lesions being caused by an organism of the streptothrix or xocardia group could not be excluded. From the evidence obtained it would appear that inhalation is the common method of infection, such infection arising from dust- infected particles. Once tuberculosis is established in the lung, dust appears to be an irritating factor favouring the progress of the disease ; incidentally it may be mentioned here that the practice of recommending cases of early tuberculosis to a country such as the Sudan is one to be deprecated inasmuch as they invariably become worse, With regard to sex and age, the disease appears to be more prevalent amcng adult males, but allowance should be made for the fact that racial customs, more especially in some parts of the Sudan, do not encourage the female population to seek medical advice; however, having due regard to this, it would appear that the disease 1s more prevalent among the itinerant male population, a fact which is not in accordance with Lankester’s (1920) observations in India. The children of the Sudanese appear to be rarely affected. Of the varieties of tubercular disease in the Sudan, adenitis is perhaps the commonest; with lung tuberculosis, and a pleurisy of tubercular origin next in frequency ; general miliary tuberculosis also occurs probably more commonly than is suspected, presenting with its pyrexia, cachexia, and splenomegaly, a clinical picture often 239 difficult of diagnosis and readily confused with other diseases. Pott’s disease 1s exceedingly rare, and tubercular meningitis more so. It is doubtful whether skin tuberculosis exists. Cases labelled as such have, on bacteriological examination, proved to be early tubercular leprosy. A few cases of joint tuberculosis have been observed by the writer, but are uncommon. Intestinal tuberculosis occurring as a primary affection of the intestines is exceedingly rare, as would be expected in a country where animal tuberculosis is non-existent. Recently a case of breast tuberculosis came under the writer’s observations, and as the disease is of sufficient rarity even in Western countries, a few detailed notes regarding this case are appended. The patient was a Sudanese woman, about 4o years of age, hailing from the remote hilly districts of Kordofan, where she had spent the greater part of her ‘life. She was married, and had a grown-up daughter who was in good health. According to her statement, her Ulness commenced some sixteen months ago with a painful swelling of the breast, which was not attributed to any injury received. The symptoms lasted for a period of twelve months and then subsided ; however, about three months ago, she had recurring attacks of pain, and decided to come to Khartoum for treatment. On admission to hospital her general condition was good, and during the few days prior to operation she showed a slight rise of temperature in the evenings. On examination of the affected left breast, there was apparent a marked retraction of the nipple (Plate XII, fig. 1), but no evidence of ulceration or scar formation. On palpation, a nodular condition of the breast was detected. The nodules appeared to be located in the breast substance, were firm in consistency, and freely movable over the subjacent muscle tissue. The axillary lymphatic glands on the left side showed no appreciable enlargement, and were painless on palpation. The right breast appeared to be perfectly healthy. Apart from the breast pain, the patient complained of no other symptoms. Examination of the lungs, heart and abdominal viscera revealed no abnormalities, nor were any enlargements of the cervical, subclavicular, mesenteric or groin glands detected. A total excision of the left breast was carried out, and some enlarged lymphatic nodes encountered during the operation were cleared away. 240 On sectioning the breast, numerous greyish-white, irregular- shaped nodules of various sizes were found scattered throughout the breast tissue (Plate XII, figs. 2 and 3). At the base of the nipple many of these nodules had coalesced and appeared to be fibrous. The majority of the nodules were firm in consistency ; some, on the other hand, had broken down to form soft caseating masses, which could be readily shelled out of a capsule composed of dense fibrous tissue. Subsequent histological examination of some of these nodules revealed their lymphatic structure. The gross pathological appearances of the breast suggested tuberculosis in which fibrosis was a marked feature. Film preparations of the broken down connecting debris were stained for the purpose of demonstrating tubercle bacilli, but with negative results. Portions of the nodules with adjacent breast tissue were excised, fixed, embedded and_ sectioned for histological examination. Sections showed almost a complete absence of normal breast tissue. Necrotic foci of various sizes composed of granular amorphous material in which only a few nucleated cellular elements could be seen were scattered throughout the section. The larger foci, repre- senting advanced caseous degeneration, were sharply demarcated by a zone of dense fibrous tissue. The smaller foci showed a pericellular reaction composed chiefly of lymphocytes and connective tissue cells, while scattered irregularly throughout the tissue were giant cells of Langerhams, containing six or more nuclei (Plate XII, fig. 4). The blood vessels showed a periarteritis and also some thickening of the tunica media. Sections of the lymphatic nodules showed well marked caseation with separative fibrotic changes and typical giant cell systems. The vessels here also showed a periarteritis and mesarteritis. Sections of the nodules were also stained by special methods to demonstrate tubercle bacilli, but with negative results. REMARKS There is little or no doubt that the case represented one of tuberculosis of the breast in which reparative changes of a fibrotic nature were a feature. Such changes probably accounted for tubercle 241 bacilli not being found in the sections and film preparations, and were also responsible for the marked retraction of the nipple. It is to be regretted that no inoculation experiments were carried out, but in view of the reparative changes noted it is doubtful whether they would have led to a successful issue. In all probability, the breast was secondarily infected via the lymphatics, although no primary focus of infection could be detected. In view of its rarity, even in Western countries, the case appears worthy of record, and no similar case appears to have been previously reported from the Sudan. I am indebted to Dr. Hodson, M.V.O., Director, Khartoum and Omdurman Civil Hospitals, for furnishing the clinical notes of the case and for providing the material for examination. KHARTOUM, June 1, 1922. REFERENCES Arcursatp, R. G. (1910). Acid-fast Bacilli in a Camel’s Lung, the gross lesions of which closely simulated Miliary Tuberculosis. ‘fourn. Comp. Path. G Therapeutics. Vol. XXIII, p. 56-57. Batrour, A. (1904). First Report of the Wellcome Tropical Research Laboratories. Busunett, G. (1920). A study in the epidemiology of Tuberculosis with special reference to Tuberculosis in the Tropics and of the Negro Race. Cummins, S. L. (1920). Tuberculosis in primitive tribes and its bearing on the tuberculosis of civilized communities. Internat. Fourn. Public Health. Exxior Situ, G., and Woop Jones, F. (1907-1908). Archaeological Survey of Nubia. LanxesteR, A. (1920). Tuberculosis in India, its Prevalence, Causation, and Prevention. Morea, L. (1919). Fréquence de la tuberculose pulmonaire chez les représentants des races colorées importés en France. Observations radiologiques. Bull. Acad. Med. Feb. 25. Rovusier, C. (1920). Les formes cliniques de la tuberculose thoracique chez les troupes exotiques importées en France pendant la guerre. Gaz. des Hopit. Oct. 2. Rurrer, Sir A. (1921). Studies in the Palaeopathology of Egypt. Edited by Prof. Moodie. Scorr, H. (1921). The Prevalence and Character of Tuberculosis in Hong-Kong. Annals Trop. Med. & Parasit. Fig. Fig. 242 EXPLANATION) OF, PEATE, oxi Anterior view of the affected breast showing in the centre the marked retraction of the nipple. Section of the same breast showing the tubercular nodules demarcated by fibrous tissue. Showing a large caseating lymphatic node at the breast margin. ; Microphotograph of a section showing a single tubercle. In the centre is a giant cell sending protoplasmic processes into the surrounding epithelioid cells. The marginal portion of the tubercle shows the lymphoid cell infiltra- tions es70: XII PLATE Annals Trop. Med. & Parasitol., Vol. XVI Fics I 4 ‘ ™ my PA ON Ltd., Imp. e S Co., T inling G 243 WEST AFRICAN CERATOPOGONINAE PART II BY A. INGRAM AND Ton Wee Sa MACEIE. (Received for publication 28 June, 1922) The majority of the species described in this paper were collected at or near Accra, in the Gold Coast. A few, however, were sent to us from Nigeria, and for these we have to thank Dr. E. C. Braithwaite, of Calabar, and, once again, Dr. H. Andrew Foy, of Lagos. With regard to the species which in this and our previous papers we have assigned to the Genus Aévichopogon, it should be noted that the eyes are not bare. In his paper on the ‘Chironomidae of America’ (1917), Kieffer associated his Genus Kemfia with Atrichopogon, separting it by the pubescence of the eyes. Later (1921), in a brief note, the same author erected the new Genus Gymnohelea, the characters of which also agree with those of A ¢richo- pogon excepting that the eyes are pubescent. Kieffer appears therefore, to recognise two genera (Kempza and Gymnohelea) closely allied to Advichopogon but differing from it in having the eyes pubescent, but he has not stated what are the differences between them, nor indeed, so far as we can ascertain, has he fully detailed the generic characters of either. In the species which we have . described, we have found every gradation between those in which the eyes are practically bare and those in which they are densely hairy. In the former, the pubescence may be restricted to the ’ anterior margins of the middle thirds of the eyes and may be visible clearly only after treatment with caustic potash, so that it might be overlooked (as was done by us in some cases) unless, by rolling the specimen from side to side as is possible by our carbolic 244 technique, the whole eye were carefully examined.* In our opinion, the hairiness of the eyes cannot, therefore, be considered of more than specific value, and accordingly we have referred our species to the Genus A?richopogon. The species which on a previous occasion (1921) we described as Kem pia ochrosoma should also, we consider, be referred to this genus. The figures illustrating the specific descriptions are in most cases mere outlines, drawn with the aid of a camera lucida, omitting such structures as the hairs on the hypopygium, and the fringe and the stronger setae on the costa and basal veins of the wings. The unit of measurement referred to is 3°8p. The types and co-types of the new species described have been deposited in the Museum of the Liverpool School of Tropical Medicine. Thysanognathust (Prionognathus) albopictus, sp. nov. MEASUREMENTS. Male. Female. Length of body| (one male and one female) .... 1-2. mm. 1-3 mm. Length of wing sa 500 500 508 .. O9 mm. o-9 mm. Greatest breadth of wing ... SSE ef .. 023mm. 04mm. Head dark brown. Eyes narrowly separated above in the female, more widely in the male; in both sexes the space between them wedge-shaped, broadest at the vertex. Clypeus and proboscis dark brown. Palpi dark brown: in the female, the second and fourth segments sub-equal but the fourth the more slender, third rather longer, slightly inflated, with a large pit containing very long sensory hairs, fifth longer than the third, slightly dilated at its end; in the male, second, third, and fourth sub-equal, third not inflated, fifth longer and slightly dilated at its end. Axdennae: in the female, first segment brown, bearing a few hairs, torus brown, rounded, bearing a few hairs; flagellum pale brown basally and ‘darker brown apically, its segments somewhat flask-shaped, all * Referring to this question Mr. F. W. Edwards has written to us as follows : ‘ Ceratopogon fusculus, Cog., which I believe is the type of Atrichopogon, certainly has the eyes entirely bare, ~ but like you, I have found species of this group which have the eyes only very slightly hairy on the upper part, and bare below. In consequence of this, I have long been of the opinion that Atrichopogon and Kempia ought to be united.’ + Mr. F. W. Edwards has kindly informed us that a new name is required for this genus as Prionognathus, C. 1. and M. is preoccupied. See Scudder’s Nomenclator. In all cases this measurement is taken from the anterior margin of the thorax to the tip of the abdomen of specimens mounted in carbolic. 245 about twice as long as broad, and forming a continuous series from base to apex, the last segment being slightly longer and broader, not flask-shaped, ending bluntly without a stylet. In the male, first segment a mere ring of chitin; torus brown, large, bearing a few hairs; flagellum pale brown basally, bearing a pale brown plume, and dark brown distally, the twelfth segment slightly produced distally, length about twice the breadth, the last three segments elongated, nearly five times as long as broad, the last segment ending bluntly without a stylet. Zorax dark brown with pale, almost silvery, markings. Dorsum dark brown, with a broad median pale stripe, and on each side of it two small pale spots anteriorly and a larger, more diffuse pale area at the root of the wing. These pale markings are larger and more distinct in the male than they are in the female. Pleurae brown. Scutellum greyish-brown, containing an almost white pigment, bearing in both sexes four central bristles, two anterior and two marginal, the latter close together. Post- scutellum dark brown with two large pale, grey, patches anteriorly. Wings (fig. 1) hyaline, with two small, blackish spots, one covering Fic. 1. Thysanogaathus (Prionognathus) albopictus, sp.n., wing of female to show distribution of decumbent hairs, adornment, and venation. c. go. the extremities of the costa and first and third veins, the other smaller about the middle of the fused portion of the first and third veins, covering only the anterior half of the vein. The anterior half of the basal portion of the wing proximal to the anterior cross-vein 1s also infuscated. These dark markings are much paler and less distinct in the male. Decumbent hairs fairly numerous in the female on the distal third of the wing at the tip, between the rami of the fourth vein, and (to a lesser extent) between the fourth and fifth veins; in the male they are entirely wanting. Halteres with greyish-brown, 246 almost white, knobs, paler in the male than in the female. The knobs contain a whiteish pigment similar to that present in the scutellum and in the abdomen. Legs in the female darkish brown, with pale bands; femora with pale sub-apical bands, tibiae with pale sub-basal bands, and on the middle and hind legs pale sub-apical bands also, first three tarsal segments pale, with slightly infuscated apices excepting the first tarsal segment of the hind legs which is entirely dark brown, fourth and fifth tarsal segments of all iegs infuscated ; in the male the legs are much paler, but similarly marked. Claws in the female unequal, one very large, about as long as the fifth tarsal segment, the other small, on the middle and hind legs about a quarter the length of the segment, on the fore legs longer, about half the length; in the male, claws equal, small, less than half the length of the fifth tarsal segment, with bifid tips. Aédomen dark brown with pale grey, almost silvery, markings on the sides and posterior margins of the segments. Jn the female the pale markings are somewhat broken up into small spots, and are most conspicuous on the fourth to the sixth segments; the tip of the abdomen is white. In the male the pale markings are larger or smaller marginal patches on each side of the middle line, and are most conspicuous on the fifth to the seventh segments; the tip of the body (excluding the claspers) is dark brown. Spermathecae two, highly chitinised, more or less pyriform and slightly unequal; in one specimen the lengths and breadths were about 80” by 75u, and 65 by 6S» respectively. The chitinised parts of the ducts in the same specimen measured about 10 and 8» respectively, but as they merged insensibly with the bodies of the spermathecae the measurements are not exact ones. HypopyGium (fig. 2). Highly chitinised, dark brown, excepting the claspers and the posterior end of the ninth tergite. Nznih segment: tergite long, broad and highly chitinised at the base, narrow and feebly chitinised at the apex, very sparsely clothed with hairs dorsally, the posterior margin straight, without either notch or lateral finger-like processes; sternite very deeply and widely excavated. Forceps: side-pieces well developed, highly chitinised, tapering distally; claspers very feebly chitinised and very pale coloured, curved slightly inwards, covered all over with minute hairs, and with the ends divided into two small processes, the ventral 247 one the larger and spine-like. Harpes very highly chitinised, distal portion directed posteriorly, broad at the base, tapering towards the ‘) B Fic. 2. Thysanognathus (Prionognathus) albopictus, sp.n., male hypopygium. a—ninth tergite, dorsal view ; b—ninth sternite, side pieces, and aedoeagus, ventral view ; c—aedoeagus and harpe, lateral view; d—harpe, ventral view ; e—clasper, lateral view. All x c. 375. apex, and ending in a short hook. Aedoeagus with a very wide basal arch and a long posterior process which is bent sharply at its 248 end in a ventral directicn. Membrane joining the aedoeagus to the ninth sternite covered all over with spicules. GOLD Coast: Dodowah, 18th February, 1922; two females and one male, reared from material taken from a rot-hole in a mango tree. Thysanognathus (Prionognathus) melanostictus, sp. nov. MEaAsuREMENTS. Length of body (one male) a or whe ak .. TOmm. Length of wing “ie sae S00 aa ise sr ... Og mm. Greatest breadth of wing ... see sae oar oss 2 O-3emaims Head brown. Eyes separated, the space between them being wedge-shaped, broadest at the vertex. Clypeus, proboscis, and palpi brown. Second, third, and fourth palpal segments sub-equal, fifth longer, slightly dilated distally. Aw¢exnae: torus very large, brown; flagellum unfortunately missing. Zovax greyish-brown with small dark brown spots. Scutellum greyish-brown, dark brown mesially, bearing four central bristles, two anterior and two marginal, the latter close together. Post-scutellum dark brown with two large pale grey areas anteriorly. Pleurae brown. Wzxgs hyaline with small blackish spots as shown in the figure (see fig. 3). _Decumbent Fic. 3. Thysanognathus (Prionognathus) melanostictus, sp.n., wing of male to show adornment and venation. X c. 90. hairs practically absent, only one or two being present at the periphery near the tips of the wings. Halteres with greyish-brown, almost white, knobs. Legs greyish-brown with dark markings similar to those of P. marmoratus. Claws similar to those of P. marmoratus. Abdomen dark brown with pale grey markings. Hypopycium (fig. 4). Somewhat similar to that of P. marmoratus, not very highly chitinised. Nnth segment: tergite moderately long, feebly chitinised especially posteriorly, dorsal surface with six 249 long, strong hairs, three on each side in an oblique row, posterior margin rounded, not notched, with the lateral finger-like processes reduced to small elevations each bearing a short hair; sternite deeply excavated in the middle line posteriorly. Forceps: side-pieces well developed, highly chitinised especially at the base; claspers long, poorly chitinised, of almost uniform width throughout, basal three- quarters clothed with minute hairs. Harpes highly chitinised ; basal Fic. 4. Thysanognathus (Prionognathus) melanostictus, sp.n., male hypopygium. A—ventral view; B—lateral view of aedoeagus and harpe. X c. 375. portion directed laterally; distal portion a long, almost straight, rod-like structure directed posteriorly, reaching to the posterior margin of the ninth tergite, and tapering to a point. Aedoeagus highly chitinised basally, curved, shaped as shown in the figures. Membrane connecting the aedoeagus to the ninth sternite covered with spicules. GoLp Coast: Accra, February, 1922; a single male, collected upon a window in the laboratory. Dasyhelea flavipicta, sp. nov. MEASUREMENTS. Length of body (two pas Bes se a a .. O gmm. Length of wing... x Se its Sc Be .. O 7mm. Greatest breadth of wing ... ‘x x re bes ... 026mm. Head brownish-yellow, the middle of the occiput brown. Eyes narrowly separated. Clypeus brownish-yellow. Proboscis and 250 palpi brownish-yellow; third segment of palp cylindrical, not inflated, nearly as long as the fourth and fifth segments together, sensory hairs very few. Antennae: torus dark brown, flagellum paler brown, with conspicuous short and long spines on all the segments, and rather short brown hairs, First segment small, hair- less ; segments fcur to ten oval to elongate-ovoid, slightly constricted at the apex but not flask-shaped, the length varying from one and three-fifths to one and four-fifths the width; segments eleven to fifteen slightly longer and more flask-shaped, length from about twice to two and a half times the width, the last segment not ending in a stylet but tapering to a blunt pomt. Torax bright yellow with dark brown dersal bands similar to those of D. flava but darker and not distinctly separated. Scutellum (fig. 5) bright yellow, very slightly darker at the sides, with two lateral and four centro- marginal bristles and a single central-sub-marginal, small hair. Fic. 5. Dasyhelea flavipicta, sp.n., scutellum of female. x c. 375. Post-scutellum dark brown. Pleurae yellow or yellowish-brown. Wings clear, without spots, rather thickly clothed with long decumbent hairs which extend to the base between the fourth and fifth veins. The bifurcation of the fourth vein before the middle of the wing, that of the fifth vein at about the same level as the end of the costa. Halteres with bright yellow knobs and brownish stems. Legs almost uniformly light brown; claws short, equal, simple. Abdomen: dorsum dark brown, venter paler, with yellow pigment (soluble in caustic potash) visible laterally and, when the abdomen is distended, between the segments. Spermathecae similar to that of D. flava, single, highly chitinised, pyriform, length about 45; the commencement of the duct is chitinised. Chitinous plates on the ventral aspect in the neighbourhood of the vulva unlike those of D. flava, and the tubular process present in that species apparently not developed (fig. 6). 251 ~six Oe within svint Sere awe Fic. 6. Dasyhelea flavipicta, sp.n., posterior extremity of abdomen of female. d—lateral view ; B—ventral view. xc. 280. i—tergite; s—sternite ; c—cerci. GOLD Coast: Accra, 8th March, 1920; collected in the evening upon windows in the laboratory. This species closely resembles D. flava, but is darker, and differs from it in the form of the antennal segments as well as in other respects. Dasyhelea omoxantha, sp. nov. MEAsuREMENTS. Female. Male. Length of body (one specimen of — .. 1:2 mm. I-2 mm. Length of wing... 35 Sea .. O9mm. o-9 mm. Greatest breadth of wing ... on see seo (Oley 0-3 mm. Head dark brown. Eyes narrowly separated in both sexes. Proboscis dark brown. Palpi paler brown; fifth segment swollen at the end, fourth slightly shorter than the fifth, third shghtly longer and only feebly inflated in its lower half. Aztennae dark brown, bearing dark brown hairs and, on segments four to ten of the flagellum at least, short and longer, curved, spines; in the female, segments four to ten sub-spherical to ovoid, length from a little over once to once and two-thirds the width, segments eleven to fourteen rather more elongate, length from a little over once and two-thirds to nearly twice the width, the last segment broad, without a stylet ; in the male, segments four to eleven ‘spheroidal to ovoid, segments twelve to fourteen elongated, sub-equal, about three times as long as broad, binodose, the last segment slightly shorter, broader at the base and tapering to a conical end without a stylet. Thorax dark 252 brown with large, yellow, humeral patches. Scutellum almost entirely yellow, but slightly darker at the sides, bearing in both sexes two lateral and three centro-marginal bristles and no small hairs. Post-scutellum dark brown. Pleurae yellow above, dark brown beneath. Wzzgs without spots. Decumbent hairs in the female fairly numerous and extending as a row aimost to the base between the fourth and fifth veins, in the male fewer, not extending basally beyond the level of the cross-vein, and absent from the anal angle and the fork of the fifth vein. Costa not reaching as far as the middle of the wing in either sex; terminal cell well developed in the male but almost obsolete in the female. Fork of the fourth vein proximal to the middle of the wing in both sexes, that of the fifth vein in the female at about the level of the end of the costa, in the male slightly more distal. Halteres with pale yellow knobs. Legs brown, often a reddish colour, the proximal segments and the joints slightly darker; claws small, simple, equal, with a slight basal extension, and in the male with bifid tips. Aédomen dark brown, venter paler than dorsum. Spermatheca single, highly chitinised, pyriform, length 32, greatest breadth 27, the duct chitinised for only about 2» or less at its commencement. Hypopycium (fig. 7). Nznth segment: tergite broad, tapering only slightly, and scantily clothed with long, dark hairs, especially A Fic.7. Dasybelea omoxantha, sp.n., outlines of male hypopygium, ventral views. 4—ninth segment, forceps, and aedoeagus; B—harpes. X c. 375. 253 in the middle line on the posterior quarter, posterior margin not notched, lateral angles squared, each with a single small hair and, more ventrally, a small process directed somewhat inwards and covered with short, stiff, hairs; sternite apparently prolonged posteriorly in the middle line as a delicate, more or less conical process. Forceps: side-pieces well developed, hairy; claspers single, rather highly chitinised; pubescent all over, and bearing in addition a few larger hairs at the base and apex. Harfes: basal portions very unequal and highly chitinised, appearance very variable in different specimens; from the right basal portion arises a long posterior projection which ends in a long pointed tip. Aedoeagus forming a broad, highly chitinised arch with a short posterior projection on each side. NIGERIA (Southern Provinces): Calabar, February, 1922; one female and three males (Dr. E. C. Braithwaite). This species resembles in some respects both D. luteoscutellata and D. incon- Spicuosa, especially the former, but may be distinguished by the colour of the halteres and by the large, yellow, humeral patches. The hypopygium of the male is characteristic. Atrichopogon africanum, Ingram and Macfie MEAsurREMENTS. Length of body (one ete 4 id oe ae eee? Tom. Length of wing... ve cr Sie be sis aco, ME gyautiael Greatest breadth of wing ... ii mee a ae ... O4mm. Head dark brown, clothed with dark brown hairs. Eyes pubescent, rather sparsely and apparently only on the upper halves; contiguous but with the facets narrowly separated. Clypeus, palpi, and proboscis dark brown. First palpal segment small, second about as long as the fifth, third longer, somewhat inflated, and bearing a well developed sensory cup at about its middle, fourth about half the length of the third, and fifth rather longer than the fourth and only slightly expanded at its end. Axéennae: first segment and torus dark brown, the former a mere ring of chitin, the latter large, very dark, hairless. Flagellum paler brown, the terminal segments somewhat darker than the rest, with a well developed plume of brownish hairs. The twelfth segment is slightly prolonged at its distal end, length rather more than four times the 254 breadth, the last three segments elongated, about seven times as long as broad, the fourteenth being the shortest and the fifteenth the longest and ending in a long (about 20), pointed, stylet (fig. 8a). The combined lengths of segments twelve to fifteen is rather greater than the combined lengths of segments four to eleven, namely (excluding the stylet, which measures about five units) 125 to 111, or 112 to 1. Thorax uniformly dark brown. Scutellum dark brown, bearing two admedian-and two lateral bristles and Cc B A Fic. 8. Atrichopogon africanum, I. & M., Ad—outline of the last segment of the antenna of the male; B—hypopygium of male, aedoeagus in lateral view ; C—side-piece to show the dorsal root-like process. All X c. 375. about six small hairs. Post-scutellum dark brown. Pleurae dark brown. Wings clear, unspotted; surface covered by microtrichia but without Jonger decumbent hairs. Venation as in A. (K.) ochro- soma. Halteres with rather dark brown knobs. Legs almost uniformly brown, tarsal segments rather darker than the others, unarmed. Claws equal, small, about half the length of the fifth tarsal segment, bifid at the tips. Empodium well developed, hairy, 255 at least as long as the claws. Addomen dark brown, but not so dark as the thorax. Hyvopycium (fig. 8, B and C, and fig. 9). Generally similar to that of A. (K.) ochrosoma. Ninth segment well chitinised : tergite long, bearing (especially on its posterior fourth) a number of strong hairs, posterior margin rounded, without lateral, finger-like processes; sternite deeply notched, bearing a few hairs. Forceps highly chitinised, normal in form; side-pieces with large, curved, dorsal root-like processes which articulate with the proximal ends of the aedoeagus ; claspers rather strongly chitinised, entirely covered Fic. 9. Atrichopogon africanum, I. & M., part of male hypopygium, ventral view, showing the ninth sternite and the aedoeagus. x c. 375. ; by pubescent hairs, and bearing in addition a few longer hairs. Harpfes apparently wanting. / 073 mm, Head dark brown. Eyes sparsely hairy, the pubescence, as in A. kelainosoma, most distinct at the sides near the anterior margin; contiguous but with the facets narrowly separated. Clypeus, palpi, and proboscis brown. Second and fourth palpal segments sub- equal, third small, only slightly longer than the fourth (twelve to nine units), slightly inflated, and with a small sensory pit. Antennae: first segment a mere ring of chitin; torus large, dark brown, bearing about six hairs; flagella unfortunately missing. Thorax dark brown. Scuatellum darkish brown, bearing two lateral and two admedian bristles and no small hairs. Post-scutellum dark brown. Pleurae brown. Wéngs clear, unspotted, without decumbent hairs; venation as in A. africanum. Ualteres with white knobs. Legs almost uniformly brown; claws unfortunately missing. Abdomen darkish brown. Hypropycium (fig. 15). Ninth segment well chitinised: tergite moderately long, sparsely clothed with hairs, posterior margin rounded, without lateral finger-like processes; sternite not notched, Fic. 15. Altrichopogon hesperium, sp.n., outline of male hypopygium, ventral view. X C. 375+ 267 but with a group of a dozen stout setae in the middle line posteriorly, Forceps normal, dorsal root-like processes of the side-pieces long, claspers short, with broad, hairy bases and strong, sharply pointed, almost claw-like extremities. Aedoeagus rather highly chitinised and characteristic in form (see figure). GOLD Coast: Accra, 24th December, 1921; collected in the evening upon a window in the laboratory. This species is described here, although at present we possess only a single, damaged specimen, on account of the characteristic form of the hypopygium. Stilobezzia limnophila, sp. nov. MEASUREMENTS. Length of body (one male) sat oe or soc ... O° 9mm. Length of wing... ae oa ee: Avs ssa ... Oo: 8mm. Greatest breadth of wing ... : 0-23 mm. A small, delicate, greenish-brown midge. Head dark brown. Eyes bare, contiguous above but with the facets rather widely separated. Clypeus, proboscis, and palpi brown. Second and third palpal segments sub-equal, third not inflated and without a sensory pit, but with a small anterior depression from which arise a few sensory hairs, fourth slightly shorter than the third, cylindrical, fifth longer than any of the other segments, widely dilated at its end, pyriform. Aztennae: first segment moderately large, brown; torus large, rounded, yellowish-brown, bearing a few hairs; flagellum unfortunately missing. Tfovax almost uniformly dark greenish- brown. Pleurae dark greenish-brown. Scutellum dark greenish- brown, bearing two admedian and two (smaller) lateral bristles, but no small hairs. Post-scutellum dark greenish-brown. Wemgs clear, without dark markings; surface covered by microtrichia but without longer, decumbent hairs. Venation as in S. spivogyrae. Halteres _ with dark, greenish-brown knobs. Legs long, almost colourless, but the knees and the apices of the tibiae are slightly yellowish-brown. Rows of small spines on the first and second tarsal segments as in S. spirogyrae; apical pairs of spines on the tarsal segments poorly developed, and other spines apparently absent. Third tarsal segment on all the legs bell-shaped, fourth cordiform. First tarsal segment of hind legs about twice the length of the second. Claws equal, small, less than half the length of the fifth tarsal 268 segment, bifid at the tips. Empodium rudimentary. Abdomen dark greenish-brown dorsally, excepting the first two segments which are pale; venter paler. Hypopycium (fig. 16). Ninth segment: tergite sparsely clothed with long hairs, rather short, tapering shghtly, the posterior margin rounded, notched, without lateral finger-like processes; sternite very Fic. 16. Stilobezxia limnopila, sp.n., hypopygium of male, ventral view. c. 375. short, slightly excavated. Forceps feebly chitinised: side-pieces sparsely clothed with long hairs, each with a broad, inwardly- projecting, basal process, and a highly chitinised, beak-like, dorsal root-like process; claspers blunt, clothed with minute hairs and a few rather longer, delicate hairs. Harvfes long, slender, highly chitinised, crossing in the middle line; basal portion foot-like, distal portion long, tapering gradually. Aedoeagus V-shaped, the membrane joining it to the ninth sternite spiculated. GOLD Coast: Accra, 26th December, 1921; reared from mud from the margin of a pool near the station for the Weshiang Railway. Monohelea nigeriae, sp. nov. MEASUREMENTS. Length of body (one saan os as es as 2) mim: Length of wing ose a Sec ae 308 Ss .. o8 mm, Greatest breadth of wing ... ee 5 408 a sh .. O3 mm. This insect is generally similar to Monohelea litoraurea. Head dark brown. Eves above widely separated posteriorly and just 269 touching anteriorly. Clypeus, proboscis, and palpi dark brown. Stylets of the proboscis highly chitinised, mandibles strongly serrated at their ends. First palpal segment very small, second and fourth sub-equal, third about once and a half the length of the fourth, inflated about its middle, bearing a large sensory pit, fifth slightly longer than the fourth, somewhat dilated at its end. Axtennae brown: first segment rather large, darkish brown, bearing a few hairs; torus rounded, rather dark yellowish-brown, bearing a few hairs; flagellum paler brown, the distal halves of the basal segments and the whole of the last five segments darker. Segments four to ten almost sub-equal, about twice as long as broad or a little longer; segments eleven to fourteen elongated, about four times as long as broad, the last segment slightly longer (24 units by §), tapering at its extremity. Combined lengths of segments eleven to fifteen rather greater than the combined lengths of segments four to ten, namely, about 106 units to 93, or 1:14 to 1. Yhorax darkish brown. Scutellum darkish brown, darker in the centre and at the sides, bearing two admedian and two lateral bristles, and six short hairs. Post-scutellum dark brown. Pleurae dark brown. Waemgs (fig. 17) Fic. 17. Monobelea nigeriae, sp.n., wing of female (fringe not shown). c. go. brownish, markings somewhat similar to those of 4. litoraurea, surface covered by microtrichia but without longer, decumbent hairs. Venation as shown in the figure. Halteres with dark brown knobs and pale stems. Legs: coxae, trochanters, femora, and tibiae on all the legs dark brown, with the exception of the knees which are pale, yellowish; tarsal segments pale brown, the first tarsal segments of the hirid legs, however, rather darker than the others. Form of 270 the segments and armatures of spines as in M/. litoraurea; fourth tarsal segments cylindrical. First tarsal segment of the hind legs two and a half times as long as the second, and with a double bend at the base. Claws on the fore and middie legs equal, rather long, about three-fifths the length of the fifth tarsal segment, each with a small basal tooth; on the hind legs single, longer (but not so long as in M. litoraurea), about as long as the fifth tarsal segment, with a basal tooth. Empodium rudimentary. Addomen grey-brown with dark markings in the fresh state, almost uniformly dark brown after preservation; tip of the body pale, almost white. Spermathecae two, highly chitinised, oval or sub-spherical, unequal, diameters in the single specimen examined approximately 57” by 45m, and 50m by 50. the commencement of the duct chitinised for only a very short distance. NIGERIA, Southern Provinces: Lagos, 26th November, 1921 (Dr. H. Andrew Foy); collected in the evening upon a lamp-shade. Eukraiohelea foyt, sp. nov. MEASUREMENTS. Length of body (one sac 506 oe ae ee so. /a-7emme Length of wing “03 c ces 500 200 so er) OLS sane: Greatest breadth of wing ... 0 —_ se a . \ . e a —— adit to mouriehs amon Sab ory etonlionia a Z fit + IMEIGE sit dinth mits ii ‘ : Py “iis 4 i yee ioe olf ft Dade Z Ls on seis 44. Did 2 < +s be . trodzatai Inte—eng og faitieene, Slt to Wace Zs : P = \ # \ 1 s ife--. rd es - \ \ : ) . ‘ : & ’ \ 44 2," ; 7 © i iar P ko — AS he a — y ~ y, rn, } P 7 - ® } is ty, & . 7 ars bit ‘Shiea'd to Seams, OARS 1 3 “ee ‘? To ae “a Qing Ineo. 7p wt < eS sore fie e pt eeatolaine cite pst(eqpay ant 125 Sette Fos ett’ aoe Loney rit jit repre h fer tonnet wis b c: Pudi ook Soptads OE glee AGRE ae ath 3 315 MOSQUITOES COLLECTED IN THE MANAOS REGION OF THE AMAZON BY R. M. GORDON AND A. M. EVANS (Received for publication 11 August, 1922) PLATE XIV The culicidae recorded below were collected by one of us (R. M.G.) at Manaos Amazonas during 1921 and the beginning of 1922. A few of the species were taken in the town or its outskirts, but the great majority were obtained in the forests surrounding Macapa, a small saw-mill about fifteen miles from Manaos on the Rio Negro. In this region only a dim light is present in the deeper parts of the forests. Here certain mosquitoes bite freely at all hours of the day, so a good deal of collecting was done by the party walking in single file, each individual ‘ bottling’ mosquitoes as they lit on the person in front. Owing to lack of proper lighting facilities, little or no work could be done at night. The breeding-places of these forest mosquitoes were difficult to locate, open pools are rare in the forest, and almost devoid of larvae when found. The most common breeding-places encountered were (1) reservoirs of water in natural crevices in the bark of trees; (2) rot-holes in trees ; (3) water reservoirs in plants. The food supply of these mosquitoes is doubtful, their chances of biting man are negligible, and animal and bird life seems extremely scarce. Particular attention was paid to searching for Anophelines, none were discovered in the forest, the only ones recorded Anopheles (Cellia) albimanus being taken in the town or outskirts of Manaos. Newstead and Thomas (1910) suggested that it was ‘highly probable that other mosquitoes await the discoverer in a region so rich in insect life .’; among the present collection are many 336 species not recorded hitherto, of which four are new and two appear to be well marked varieties of existing species. Sabethes amazonicus, sp.n. FeMALe. Head. Proboscis long and slender, gradually enlarged apically. Clyfeus and forz black with grey pruinosity. Scales of occiput with deep blue, violet and green reflections‘ above, white beneath. Prothoracic lobes covered with metallic scales with bright blue and green reflections varying according to the light; a row of coarse black bristles along the margin. Mesonotum largely denuded, the scales present similar to those of prothoracic lobes. Scztellum with lateral lobes metallic green scaled, mid lobe denuded. Aletanotum with four coarse black setae. Pleurae and coxae with flat white scales. Abdomen: Tergite of first segment with bright metallic green scales; white at sides. Scales of rest of tergites metallic with deep blue, pale blue, and green reflections according to the direction in which they are viewed. Sternites white scaled. Wings with strong reddish-brown infuscation. Scales on knobs of halteres metallic yellowish-green. Legs long and slender. Hind legs with paddles of long, out- standing scales involving distal half of tibia, metatarsus, and most of second tarsal segment; the longest scales about 1'2mm, _ Front legs with tufts of outstanding scales on the distal half of the tibia and a few slightly raised scales at base of metatarsus; longest scales of tufts about o'5 mm. Hind legs entirely without raised scales. Vestiture dark brown, with bronzy, coppery and violet reflections, femora, tibiae and metatarsi without white. Front tarsi with segments three. four and basal third of five white ventrally, segment four with dark spot at middle; mid tarsi with second, third and basal half of fourth segments white all round, except narrowly at the joints. Hind tarsi with segments three, four and five ventrally white, except narrowly at the joints. Length: c. 70mm. Wing: c. 5°5 mm. Type. One female taken about three hundred yards deep in the forest, Macapa, 22nd December, 1921. 317 This species evidently comes very near S. farvsopus, D. & K., with which it agrees in having tufts of outstanding scales on the front and mid legs only. It differs from that species in the entire absence of white scales on the femora and tibiae and in the details of the tarsal markings. Sabethoides nitidus, Theob. Two larvae taken from a rot-hole in a ‘ Breau’ (native name) tree in the forest near Manaos were brought to JLiverpool alive. They were kept in an incubator at a temperature from 70° to 80° F., and one of them pupated, the pupa giving rise seven days later to a female Sabethoides. Although the specimen differs in certain details of coloration from Theobald’s (1901), Howard, Dyar and Knab’s (1915), and Dyar’s (1919), descriptions of S$. zzézdus, it is referred to this species in the absence of male specimens from this region. The specimen is more brightly coloured than typical S. nitzdus, many of the head scales having brilliant pink and imavve tints, the scales of the prothoracic lobes and mesonotum are brilliant peacock- blue, not greenish-blue as in S. zz/¢dus. The abdomen (fig. 1), seen A.M.E. Fic. 1. Sabethoides nitidus, Theo., from Manaos region, female abdomen from above. X C. 30. 318 from above, is coppery with violet reflections, and there are irregular basal patches of brassy scales on segments three and seven; broad, paired, dorsal patches, almost united in the middle, on segments four and five, and on segment six a complete broad, basal band. These brassy scales are quite conspicuous to the naked eye. Lateral basal white spots only present on last segment, not in all segments as in S. 27/2zdus. The mid legs are white scaled above on the apical three-quarters of segment two and on segment three, four and five, except the extreme tip of five. The larva of this species does not appear to have been described hitherto. Larva. Stage IV (fig. 2). Head broad. Mental plate with a my = S fF 4w = za S O-O05 millimeter F 0-05millimeter A, (C = 0-01 millimeter A.M.E. Fic. 2. Sabethoides nitidus, Theo., from Manaos region. A-E—larva; F—pupa. A—mental plate ; B—dentition of mandible ; C—spines of lateral comb (branched one in lateral view); D—siphon tube ; E—part of surface of siphon tube enlarged ; /—respiratory trumpet of pupa. large median tooth and eight smaller ones on each side. Maxillae resembling those of Sabethinus undosus as described and figured by Howard, Dyar and Knab (1915), but, left maxilla with five teeth on O-5millimeter Be mner margin, right maxilla with four teeth in this position. Mandibles similar to S. wxdosus, but dentition (fig. 2B), six (not four) teeth on a process, the terminal one large and falciform. Comb of eighth segment of twenty spines arising from membranous integument, spines thorn-shaped, some with a secondary pointed process (fig. 2C). Siphon tube three and a third times as long as greatest width, surface with groups of microscopic hairs (fig. 2 E), a row of delicate sub-equal hairs arising from posterior margin for more than two-thirds of its length. Anal segment with plate reaching about half way down segment, dorsal angle on each side with two tufts of two setae; sub-ventrally a tuft of two and a tuft of-three setae ; lateral angles of plate with a tuft of two setae at each side. Anal gills sub-cylindrical, bluntly rounded, about three-fifths as long as siphon tube. Dorsal hooks of seventh segment, if present, so small as to be undetectable in crumpled pelt. PupaA. Multiple tufts present on seventh and eighth segments. Respiratory trumpets moderately short and stout, opening wide (fig. 2F). In life, abdomen with conspicuous dark segmental bands. The coloration of this metallic scaled species appears to be extremely variable and open to a variety of interpretations. The extent of white on the hind tarsi is also subject to a considerable amount of variation in the descriptions of authors. The dorsal aspect of the abdomen was originally described by Theobald (1901) as ‘deep metallic blue with basal coppery bands’; Howard, Dyar and Knab (1915) say ‘dorsal vestiture metallic blue and green’; and Dyar (1919), in his coloration table, states that the abdomen has ‘irridescent whitish, segmental bands.’ A specimen labelled ‘S. confusus’ in the British Museum was examined by one of us (A. M. E.), and the coloration of the abdomen above was found to be dark metallic violet with scattered pale scales, and on last segments pale basal bands. Until a male from this locality is discovered, it must remain undecided whether the range of S. witidus can be considered as extending as far westwards as Manaos, or whether the genus is here represented by a distinct species. Wyeomyia negrensis, sp. n. FEMALE. Metanotum with flat white scales and a few pale setae intermixed, a tuft of dark setae posteriorly. In other respects, 320 also, closely resembling Cleobonnea occulta, B. W. and B., except that scales on disc of mesonotum broadly lanceolate. MALE. Coloration as in C. occulta, but legs differing consider- ably. Mid legs white ventrally and dark above throughout. Hind legs with femora, tibiae, metatarsi and basal quarter of second tarsal segment white ventrally; rest of tarsi brassy beneath. Hypopycium (fig. 3). Side-pieces, tenth sternites and ninth tergites as in C. occulta. Clasper with a slender, recurved, basal ‘lobe’ (Dyar, 191g) (x), with retrorse pointed tip, and a wide dilation (d@.), from which arise three lobes; an outer, rather slender lobe (2), with indications of a row of spines; a long curved lobe (3), with a row of teeth along inner side; an inner broad triangular -———_—_______| 0:05 millimeter Fic. 3. Weyeomyia negrensis, sp.n., male hypopygium, clasper. d—dilation; 1, 2, 3, 4, and 5, lobes of clasper. lobe (4), with coarse teeth along distal edge; and a secondary lobe (5), with teeth on internal surface arising from fourth lobe. Larva. Stage IV. Head wider than long, widest at posterior angles. Mental plate triangular, with a median tooth and eleven sub-equal teeth on each side. Maxilla with a terminal transparent hook-like tooth, and a row of ten transparent teeth along inner side; a sub-apical tuft of delicate hairs, and near them a single seta on a tubercle, a row of hairs internally, and a short, stout spine near outer margin. Zfovax with lateral dense tufts. Spines of comb of 321 eighth segment in a sub-triangular patch. Length of siphon tube about three times its greatest width, false pecten of four spines on distal half, three multiple tufts dorsally and a long multiple tuft ventrally at base. Anal segment with two pairs of dorsal tufts, one of five, one of two hairs; lateral hairs single, sub-ventral tufts of three and two hairs. Pupa. A tuft of two long hairs bent as shewn in fig. 4D near 0:05 millimeter /-——__ O ‘2millimeter OIL millimeter O-1 millimeter AME Fic. 4. Wyeomyia negrensis, spn. A, B, and C—larva; D—pupa. A—mental plate ; B—maxilla; C—siphon tube; D—bent hairs of cephalothorax of pupa. margin of each eye. A pair of sub-median tufts of eight branched hairs and sub-lateral tufts of four simple hairs behind insertions of antennae. Otherwise resembling pupa of C. occulta. Types. One male and one female, bred from larvae living in the stem of Bananeira braba (wild banana) in the forest near Macapa, 20th Decemer, 1921. Co-types, five females from the same source. This species is closely related to Cleobonnea occulta, B. W. and B., but there are marked differences in the coloration of the 322 legs of the male, and in the structure of the clasper. The mid legs of C. occulta are described as ‘ fale’ beneath throughout, and the third, fourth and fifth segments white above. The hind legs are described as white beneath throughout, the last three tarsal segments brassy above. The male hypopygium has the clasper with only three lobes, closely resembling, 1, 3 and 4 of W. negrensis according to Dyar’s (1919) figure of this structure; the lobes corresponding to 3 and 4 are differentiated much nearer the base than in W. negrensis; the dilation (d.) 1s absent. The branch (5) is evidently fused with the inner lobe (4) along its whole length in C. occulta. The quadrilobate con- dition of the clasper excludes W. negrensis from Dyar’s sub-genus Cleobonnea, and it is here placed provisionally in the genus Wyeomyia. Culex (Neomelanoconion) chrysothorax, Newstead and Thomas. This species was frequently taken from a pool at the Bosque, about five miles from Manaos (Plate XIV, fig. 3). Dyar (1918) suggested that it might be synonymous with C. (Choeroporpa) chrysonotum, D. and K.; and Bonne-Wepster and Bonne (1921),examined the types in the British Museum, and came to the conclusion that C. chrysothorax is a distinct species differing from C. chrysonotum ‘by the broad white apices of its femora and tibiae,’ and other characters. In view of the fact that most of the specimens in our series have the apices of the femora and tibiae only narrowly and faintly pale, the male hvpopygium was examined and compared with that of C. chrysonotum described by Dyar (1920). Five specimens were examined; they showed quite distinct differences as follows : — Z Inner branch of upper division of lobe of side-piece with larger appendage (fig. 5 Ai), a long, slender filament with recurved pointed tip, not ‘somewhat flattened and _ blade-like’ as in C. chrysonotum. Halves of mesosome (second plates, Dyar) (fig. 5 B) with a very long horn extending in the same direction as the basal hooks, arising nearer to the apex than the base; not near the base as in C. chrysonolum. It should be stated that, owing to the fact that the apical portions of the plates lie in a different plane from the basal main portion and from the horns, a considerable 323 number of totally different appearances of the whole structure may be obtained by altering the orientation (see fig. 5, CandD). In fig. 5 B the mesosome is drawn as seen when allowed to come to rest O-05millimeter bh ’ AME Fic. 5. Culex chrysothorax, Newstead and Thomas, male hypopygium. 4—clasper and lobes of side piece, 1—appendage referred to in text; B—entire mesosome, ventro-lateral view ; b.4.—basal hooks; /—line of fracture between basal hooks and halves of mesosome ; 4—horn; C and D—vyentral and lateral views of distal portion of half of mesosome. on its side; as the two halves diverge at an angle, neither half is seen in true lateral view. Culex originator, sp. n. MALE. Paltivery short, slightly less than one-sixth of proboscis, slender, pointed. Proboscis swollen distally, bent beyond middle. Occiput with pale brown, narrow curved scales in middle, and whitish scales at sides and margins of eyes. Upright forked scales numerous, black. AM/esonotum: integument dark grey, clothed with very narrow, curved, brown scales with slight greenish reflections, and numerous very long, coarse, black setae; two narrow, bare, dorsal stripes extending almost to ante-scutellar space, and a pair of wider, curved, sub-lateral bare lines extending from before wing roots outwards and backwards to lateral lobes of scutellum. Scutellum 324 O-1 millimeter AME. = Ol millimeter Fic. 6. Culex originator, sp.n., male hypopygium. 4—clasper and lobes of side piece ; i.b, and o.b.—inner and outer branches of outer division ; i.d.—inner division; B—aedoeagus, dorsi-ventral view ; b.b.—basal hooks ; b.p.—basal plate ; m—half of mesosome ; e—paramere ; St.X.—tenth sternite; J./X.—ninth tergite; J.X.—tenth tergite; ¢.1.—ventral tooth of mesosome ; ¢.p.—transparent, triangular plate ; C—half of mesosome and basal hook, lateral view ; 6.4.—basal hook ; ¢.2.—dorsal tooth ; D—tenth segment, lateral view. 4, C, and D to same scale. 325 unicolorous with mesonotum. Pleurae green, with black setae and some pale ones. Abdomen with dark brown scales above, and on segments seven and eight very pale lateral basal spots. Legs clothed with dark brown scales; femora pale beneath; hind tibiae with a line of scales beneath with brilliant yellowish, silvery reflec- tions, except on basal quarter and at distal extremity. Wings as in C. (/sostomyia) conservator, D. and K. Length: c. 35 mm. Wing: c. 2°5 mm. Hyporycium (fig. 6) Szde-piece short, rounded, width more than half the length. An area of dense setae near apex on inner side. Clasper angularly curved at right angles, gradually narrowing from bend to tip as shown in fig. 6A. Outer division of lobe of side-piece with distal half divided. Outer branch (0. 6.) bearing a large filament distally expanded as shown in the figure, and a small spine. Inner branch (Z. 6.) of outer division of lobe of side- piece bearing a stout seta at base, and a pair of expanded filaments distally, one rather more distal than the other. Inner division of lobe of side-piece (7. d.) a stout arm, exceeding the outer division, with a row of setae arising from mner side and with two rod-like appendages with curved, pointed tips, the inner situated proximal to the outer. Tenth sternites with slender stem and expanded, comb- shaped apices, with nine teeth. Aces of tenth tergites with a dense tuft of setae (fig. 5 D), the longest considerably longer than the tenth tergites. Halves of mesosome (second plate, Dyar), lateral aspect (fig. 5 C) distally pointed, with a strong, pointed tooth on upper (true ventral) edge and a blunt tooth on lower edge near basal hooks ; dorso-ventral aspect (fig. 5 B, #.), distal portion spatulate. Basal hooks well developed, strongly curved. Ninth tergites (¢. zr.) rounded, with four setae. ‘Transparent triangular plates,’ Dyar (2. p.), present between basal plates and ninth tergites. FEMALE. Vestiture similar to the male, but upright forked scales of occiput dark brown; dorsal bare lines of mesonotum partially obliterated on posterior half; faint basal lateral, pale spots on abdominal segments three to six, and apical, lateral, pale spots on segment seven. Length: c. 30mm. Wing: ¢. 2°5 mm. Larva. Stage IV (fig. 7). Dorsal head hairs consisting of an inner pair of long tufts (2. 7.) associated with a single long seta, and 326 an outer pair of shorter tufts (0. 7.). Antennae normal, spiny. Mental plate (fig. 7 B) narrow with a very wide median tooth and eight smaller ones on each side, the last one remote. Thorax rounded, wider than long. Siphon tube (fig. 7 C) very long, length nearly eleven times the average width. Pecten not reaching beyond basal quarter, three long hairs beyond. 0-O05millimeter _B. ——— O-5 millimeter pts PL DPD Ls C. AME Fic. 7. Culex originator, sp.n., larva. d—head, dorsal view; B—mental plate; C—siphon tube. Type. Male and female bred from larvae obtained from xatural holes in the bark of the ‘ Cavapana uba’ tree (native name = ‘Home of the mosquito’) about half a mile in the forest at Macapa, 21st December, 1921, emerged Ist January, 1922 (Plate XIV, fig. 2). Co-types, two males and two females from the same source, and one male from larva in rotten tree stump in forest at Macapa. The characters of the male hypopygium readily separate this species from any other described species of Culex, but outwardly it closely resembles Culex (Jsostomyia) conservator, I. and K. It differs from this species as described by Howard, Dyar and Knab 327 (1917) in having a line of brilliant yellowish, silvery scales beneath the hind tibiae, and faint pale segmental lateral abdominal spots. It would appear that the male of C. conservator has the upright forked scales of the occiput brown, not black, as in C. oviginator, and that the bare lines on the mesonotum do not extend more than half way back. Dyar (1922) discusses the hypopygial characters of C. conservator and the other two species of Culex with the male palpi as short as those of the female, C. zsostomyza bifoliata, Dyar, and C. micraedes corrigani, D. and K. From his description of the shape of the clasper in Jsostomyia, it seems probable that C. originator should be put in this sub-genus, although the divided outer division of the lobe of the side-piece, and the presence of conspicuous tufts of spines at the apices of the tenth tergites distinguish it markedly from the other two species. The latter character appears to be unique among American species of Culex. Culex corniger, Theo. A perfect female was taken in low herbage in a garden in Manaos, 7th June, -1921. Mansonia coticula, Dyar and Knab. Two females of this distinctive and beautiful species were caught about one mile deep in the forest at the saw mills, Macapa, 11 a.m. to 3 p.m., 7th December, 1921. The type specimens were taken in the Panama region, and since its discovery the species does not appear to have been found elsewhere. Our specimens, however, agree with Howard, Dyar and Knab’s (1915) description so exactly that we have no hesitation in assigning them to this species. Females of Manosnia tilillans, Walker, and M. amazonensis, Theo., were frequently taken biting man by day in the forest near Macapa. Haemagogius (Stegoconops) equinus, Theob. Three females taken in the forest near the saw mills, Macapa, II a.m. to 3 p.m., 7th, 22nd and 23rd November, 1921, are referred to this species. H. equinus has not hitherto been recorded from the Manaos region, but it has a very wide distribution in South America, Dyar (1921), and in the absence of males the present specimens must be regarded as this species. P. 327. Line 28, for Manosnia read Mansonia. 328 Psorophora lutzi, Theo. In addition to numerous females, a male of this species was caught in the forest near Macapa saw mills, 10 December, 1921. The male of P. Jutzz does not appear to have been described hitherto. It differs from the female in having yellow scales immediately in front of the ante-scutellar space, a character which O11 millimeter Fic. 8. Psorophora lutzii, Theo., male hypopygium, apex of claspette (harpagone) ; S.1, S.2, S.3 types of setae referred to in the text. was confirmed by Mr. F. W. Edwards, who kindly examined the male specimens in the British Museum collection. HypopyGium (fig. 8) with side-pieces, tenth sternites and aedoeagus as in P. posticatus. The claspettes (harpagones) H., D. o29 and K. (1917), apically expanded on inner side; with a large terminal ‘S-shaped leaf, a much smaller curved leaf, and a narrow pointed filament distally curled; internal surface with fourteen (this number may be subject to slight variation) long setae with expanded apices. The setae of three types:—I. (fig. 8, 5.1) with apices slightly swollen, bearing short simple hairs; II. (s. 2) apices considerably expanded, with longer, very delicate hairs, some of which branched ; III. (s. 3) apices produced into large membranous expanses, with fine, filamentous, branched processes. Females of this species and of P. posticatus, Wied., were the commonest mosquitoes biting by day in the forest near Macapa. Aédes (Finlaya) oswaldi, Lutz, var. brazilicnsis, n. var. Two perfect males of the /zzlaya group of Aédes were referred to this species, although they differed from it in certain respects. The differences are tabulated below :— | . A. oswaldi A. oswaldi var. braziliensis Midlegs... ...| 2nd tarsal segments with basal 2nd tarsal segment with basal third white half white. : | jae Hind legs... ...| Metatarsus with apical quarter ; Very narrow white rings at these 2nd tarsal segment with basal places third white Segment VIII of ...| Dorsally silver scaled Dorsally dark scaled abdomen The anterior three-fifths of the mesonotum are covered with very thick, bluish silvery, marrow curved scales, the whitish area being deeply incised behind. The hypopygium resembles that of A. oswaldi, but the clasper is capitate distally, not pointed as in Howard, Dyar and Knab’s (1912) figure of that species. Type and co-tyfe males bred from larvae found in hollow in tree stump, about one and a half miles deep in forest at Macapa, 8th December, 1921. 330 Megarhinus horet, sp. n. MALE. Prvodoscis about nine-tenths of the length of the wing, slender, tapering to a point; palpi slightly longer than proboscis, yvestiture of all but last segment above predominantly peacock-blue, violet towards ends of segment and in front of false articulation ; scales at dilated articulations and false articulations white, with pale mauve reflections; all but last segment with pale scales, appearing brassy or whitish according to the direction of the light. Last segment bronzy scaled with deep purple reflections. Antennae with hairs of whorls blackish-brown, second segment dotted on distal two-thirds of inner side, with metallic scales appearing peacock-blue, purple or whitish in different lights; tori black with silvery pruinosity. Clypeus short, ochraceous brown, darker in centre, with whitish pruinosity. Occzpu¢ mostly covered with olivaceous green scales, pale blue ones in front and at sides, white scales along ocular margins and beneath. Prothoracic lobes with brilliant blue scales above, violet ones towards margin, and white scales beneath, a row of coarse black setae along margin. Mesonotum, viewed without magnification from above, bronze, with a median peacock- blue stripe about one-fifth of the width of the mesonotum at the middle, posteriorly the blue area widens and coalesces with blue patches over the roots of the wings; bronze area bordered by whitish blue at edges of disc. Magnified about fifty times with binocular microscope, the bronze area seen to consist of spindle-shaped scales with brassy, coppery, greenish or light blue reflections, according to the direction in which they are viewed; the scales directed outwards - on anterior, inwards on posterior half; blue area consisting of broad, flat, backwardly directed scales, bronze when viewed from behind, metallic peacock-blue from above; pale scales bordering disc at sides and in front broader than spindle-shaped scales on disc, very transparent, whitish, with azure-blue and pale greenish-blue reflec- tions. Scales forming patches over roots of wings peacock-blue, with lighter blue and greenish reflections. Scutellum, without magnification bright metallic blue, very slightly paler than blue of mesonotum ; with magnification fifty times, scales on mid and lateral lobes similar, appearing peacock-blue with deep violet reflections, pale blue, translucent pale green or translucent brassy, according to 331 the direction of the light; mid and lateral lobes with groups of stout, black setae. Pleurae and coxae with patches of dense creamy- white scales. Spuiracular bristles seven, black; pre-alars seven, pale straw coloured ; upper mesepimerals numerous, very pale. Abdomen above, with segment one metallic pale blue, segments two, three and four with peacock-blue scales, remaining segments and side-pieces bronzy brown, with violet reflections. Sides of segments with apical patches of creamy scales, brilliant blue scales at base, some of scales with whitish and mauve reflections in certain lights. Scales of yenter creamy with silvery reflections, a median dark stripe of bronzy scales with peacock-blue reflections, lateral ciliation short, delicate, pale yellow. 0-2 millimeter AME. Fic. 9. Megarbinus borei, sp.n., male hypopygium. A—side piece ; 6.1.—basal lobe ; B—ninth tergite. Legs. Vestiture of dark scales with deep blue and purple reflections. Femora brassy beneath, knees entirely dark. Hind tarsi with fourth segment white, except at base and apex, and a very narrow line of dark scales on upper surface behind. Hypropycium (fig. 9). Basal lobe of side-piece with three stout setae at apex, of which two very long, reaching almost to insertion of clasper. Ninth tergites short, with about eleven fine setae. Length: c.10 mm. Wing: 7 mm, 332 FEMALE. Palpz: coloration above similar to male, but scales at articulations dark, with paler violet reflections; brassy scales at sides confined to basal third, rest with reddish-purple reflections. Mesonotum entirely covered on disc, except on posterior extremity, with dark bronze spindle-shaped scales, which appear deep blue with purple reflections when viewed in a direction parallel to their long axis; posterior portion between wing roots with flat scales of similar coloration. In the normal position the thorax appearing bronzy-brown, except at posterior extremity, and in irregular patches on middle regions of posterior half which appear deep ultramarine blue, owing to the antero-posterior direction of most of the scales in these regions. Laterally the scales directed more or less at right angles to the Jongitudinal axis, and, therefore, only appearing blue when the thorax is viewed from the side. Abdomen. Similar to male, but blue colour deep ultramarine, and on last two segments above an almost complete apical fringe of brassy scales. Legs. Similar to male, but mid legs with segments two and three white on anterior and dorsal surface, except narrowly at apices and bases; hind legs with segment four entirely whitish scaled, segment five with whitish scales on basal two-thirds anteriorly. LarvA. Stage IV (fig. 10). Mead, sub-quadrate, about as wide as long, insertions of antennae rather prominent, front margin deeply emarginate, produced into large prominent lobes on each side, bearing mouth brushes. Antennae cylindrical, slender, rather long, smooth, hairs sparse, internally a tuft of two hairs, externally two longer hairs on apical fourth; apex with a jointed and an unjointed appendage and a hair. Dorsal head hairs fine, three on each side behind frontal lobes, behind and internal to antennae a row of three on each side and a minute tuft internally; a single hair internal to eyes and a small branched one apparently rising from eyes. Mouth brushes consisting of nine curved blades. Labial structures (fig. 10, E, F,G) consisting of a broad chitinous fold (sub-mentum ?) hairy in middle distally with internal surface with median area heavily chitinised, tuberculate and a large stout tooth (m. ¢.) arising in centre: a mental plate attached to dorsal surface of fold (see fig. 10 G, which shows relative position of parts of labium), having a very shallow median tooth with’a small tooth on each side and Ja, aut [1W 2.9 1Omillimeter D. aN 0:2 millimeter Jayaulty[IwO.1 AME. A—head, dorsal view; B—antenna; C—mandible, to same scale as D; D—maxilla; E—labium, ventral portion ; ¢.f-—chitinous fold ; m.p.—mental plate ;_m.t.—median tooth of chitinous fold; F—secondary plate of labium; G—sagittal section of labium at X-x, $.p.—secondary plate ; H—segments VII and IX; K—respiratory trumpet of pupa. 0:2 millimeter Fic. 10. Megarhinus horei, sp.n. A-H—larva. K—pupa. 334 five large ones beyond on each side; and a ‘‘secondary plate’’ of the form shown in the figure, with the distal portion thickly dentate on dorsal surface, and the teeth tending to form a median, two lateral and inter- mediate groups. Mandible with a pair of sparsely feathered hairs (#.) on cuter side; dentition of five teeth of which two very large, ensiform ; dorsal surface with a row of short fine hairs, and a proximal row of long hairs. MVavilla rectangular, bi-lobed distally, edges of lobes densely setose, inner Icbe with a stout spine on a prominence behind insertions of hairs; outer lobe with a short, stout sensory spine rising from a tubercle almost at edge, palpi with a chitinous plate as shown in fig. 10 D, and three rudimentary jointed digits. Zo7vax rounded, the stout hairs spinulose. Addomen: lateral tufts of hairs not arising from large chitinous tubercles. Siphon tube about two and a half times as long as wide, no pecten, a single tuft near base. Large plate on side of eighth segment with two stout spinulose hairs on its posterior margin. Anal segment about as long as wide, ringed by the plate; dorsal tufts of two long brushes on each side, a single spinulose lateral hair. Anal gills very short, bud-shaped. PupaA. Respiratory trumpets as shown in figure. Length: ¢. 13 mm. Types. One male and one female, bred from Jarvae found in stems of Bananeira braba (wild banana) in the forest near Macapa, 21st December, 1921. The species is dedicated to Mr. A. T. S. Hore in recognition of valuable services, which he rendered during the collecting expeditions that were undertaken. BIONOMICS. The larvae of this mosquito were first discovered together with those of Wyeomyza negrensis, sp.n., im a stretch of forest about four miles from Macapa. As mosquitoes were extremely plentiful at this point, a small tract of forest was carefully searched for breeding-places, the larvae referred to were found by splitting up the fronds at the base of a ‘ Baxaneira braba’ (wild banana tree). As we were shifting camp the same day, the larvae had to be transported some distance in a hot sun, and none of them survived the journey. A few days later a wild banana (Plate XIV, fig. 1) was selected growing at the edge of the forest about ten miles from the spot previously examined, this was cut down close to the roots and transported to camp, where it was placed in a petrol tin, the outer 335 fronds torn off, and finally the base split up with knives. No larvae were found till the base of the tree was reached, those found were lying in the innermost fronds fully six inches from the outer circum- ference of the tree. The larvae were found to be carnivorous and had to be kept in separate tubes, where they were fed on a diet of Culex quinguefasciatus (fatigans) larvae and pupae, of which they readily destroyed two a day. In captivity they spent most of their time at the bottom of the jars, only coming to the surface at long intervals. No eggs were discovered, so the length of larval life is unknown. The average pupation period was found to be six days. Uranotaenia calosomata var. albitarsis, n. var. The specimens agree with typical U. calosomata, D. and K., in the coloration of the head, thorax and abdomen, but the front and mid tarsi have the last three segments creamy-white scaled, not as in U. calosomata, in which they are described as having ‘a brassy lustre particularly apically.’ Hind tibiae with a conspicuous bluish- white stripe extending the whole length behind; in U. calosomata the hind tibiae have only the tips narrowly silvery white. Proboscis with a bluish-white line on basal four-fifths beneath, apparently absent in U. calosomata. Hypopycium with spines on basal lobe of side-piece extending beyond the apices of the side-pieces; they are very short in Howard, Dyar and Knab’s (1912) figure of the hypopygium of U. calosomata. Type. Male and female bred from larvae taken in old iron bath at the saw mills near Macapa, 20th January, 1922; co-type, female from the same source. Other species of Uvanotaenia taken were U. geomntetrica, Theo., 3 1, flying in low herbage, Mandos, October, 1921; and U. lowii, Q 1, Manaos, 15th January, 1922. 330 REFERENCES Bonne-Wepster, J., and Bonne, C. (1919). Four new South American Mosquitoes. ns. Ins. Mens. Vol. VII, p. 105. (1921). Notes on South American Mosquitoes in the British Museum. Ins. Ins. Mens. Vol. IX, p. 20. Dyar, H. (1919). A revision of the American Sabethini of the Sabethes Group by the male genitalia. Ins. Ins. Mens. Vol. VII, p. 134, Pl. V (fig. 9). — (1920). The species of Choeroporpa, a subgenus of Culex. Ins. Ins. Mens. Vol. VIII, jBo Sys (1921) . The genus Haemagogus, Williston. Ins. Ins. Mens. Vol. IX, p. 102. ——— (1921). The species of Finlaya allied to terrens, Walker. Ins. Ins. Mens. Vol. IX, p- 152. (1922). Mosquito Notes. Jns. Ins. Mens. Vol. X, pp. 95, 96. Howarp, L. O., Dyar, H., and Kwan, F. (1912, 1915, and 1917). A Monograph of the Culicidae of North and Central America and the West Indies. Vols. II, II, and IV. Newsteap, R., and Tuomas, H. W. (1910). Mosquitoes of the Amazon Region. Ann. Trop. Med. & Parasit. Vol. 1V, No. 1, pp. 141-149. Tueoxzatp, F. V. (1go1). Mono. Culicidae. Vol. II, p. 347. ariryet! soil - hige F 338 EXPLANATION OF PLATE XIV. Fig. 1. Wild Banana (after having been cut down). Breeding-place of Megarhinus hoeri, sp.n. and Wyeomyia negrensis, sp. n. Fig. 2. ‘Carapana Uba’ Tree. Breeding-place of Culex originator, sp. n. Fig. 3. Breeding-place of Culex (Neomelanoconion) chrysothorax at Boski, Mandos. Annals Trop. Med. & Parasitol., Vol. XVI PLATE XIV C. Tinling & Co., Lid., Imp 339 TRYPANOSOMA RHODESIENSE IN A CASE OF SLEEPING SICKNESS FROM THE SUDAN BY R. G. ARCHIBALD, D.S.O., M.D. DIRECTOR OF THE WELLCOME TROPICAL RESEARCH LABORATORIES, KHARTOUM (Received for publication 25 September, 1922) Three cases of human trypanosomiasis were recently brought to Khartoum by Captain Mackinnon, M.C., R.A.M.C., Medical Officer in Charge of the Sleeping Sickness Camp at Tembura, in the Bahr-el-Ghazal Province of the Sudan. Gland puncture carried out two months previously had proved positive for trypanosomes in all three cases; mm order, however, to minimize the possibility of spreading infection during their journey through fly-infested areas, each patient had received two injections of 05 gramme atoxyl. On arrival in Khartoum gland puncture was again carried out, but trypanosomes could not be found in the several preparations examined; it was decided, however, to inoculate animals with the gland juice obtained from one of the patients. The case selected showed evidence of somnolence with a well marked enlargement of the lymphatic glands of the neck and axilla, as well as a slight degree of pyrexia. An emulsion of the gland juice with a sterile I per cent. solution of sodium citrate was prepared, and inoculated subcutaneously into three healthy gerbil rats. At the end of sixty-six days one of these rats showed an intense infection with trypanosomes in its peripheral blood; stained prepara- tions demonstrated the presence of posterior nucleated forms. Further details regarding this trypanosome and its pathogenicity for various animals will be published later by Captain Whitehead, M.C., R.A.M.C., Government Bacteriologist; suffice it to say that its morphological characters and pathogenicity for animals, justify 340 the conclusion that the trypanosome is 7. rhodeszense, an opinion shared by Professor Warrington Yorke, who kindly examined stained blood films from infected rats, as well as other data submitted. Investigations regarding the insect carrier of this trypanosome remain to be carried out; it is of interest, however, to note that Glossina fuscipes and G. morsitans are ubiquitous in the district of Tembura. The writer is indebted to the Principal Medica! Officer, Egyptian Army, for facilities granted in obtaining the material which forms the subject of this brief paper. JAMES SWIFT & SON LTD. Makers of the highest grade Microscopes and Accessories. Contractors to all Scientific Departments of H.M. Government MICROSCOPES For Pathological and Bacteriological Research. NEW SERIES OF “ TELAUGIC"’ OCULARS Giving increased Diameter and Flatness of Field, increased Brilliancy of Image, increased Distance of Eyepoint. x 7:5, x 10, x I5, X 20, X 25. Price - - £2 5s, each. HOMOGENEOUS }i-in. OBJECTIVE, N.A. 0:95 This is an entirely new objective of superlative performance, designed, in itself, tu replace both the dry 4-in. and the oil immersion ;4-in. Its slightly higher magni- fication renders it more generally useful than the Continental +-in. objectiv es. Price = - £6 15s. 81 TOTTENHAM COURT ROAD, LONDON, W.1. WATSON’S “BACTIL” MICROSCOPE A complete instrument for High Power Work, for Research, and the Laboratory. It is fitted with a Mechanical Stage of new design, giving a long horizontal traverse, Compound Substage fitted to the limb extension, and not suspended from the Stage, Body 2 in. diameter, adjusting screws to compensate for wear and tear, fitted and finished by hand, complete in case, Price: £27 10Os. Full particulars of the above, and Microscopes for all classes of work are contained in Watson’s Catalogue of Microscopes, post free on request. W. WATSON & SONS, LTD., 313 HIGH HOLBORN, LONDON, W.C.1. Established 1837. Works: Bells Hill, High Barnet. Volume XVI December 30, 1922 No. 4 ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY ISSUED BY THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE Edited by Proressor J. W. W. STEPHENS, M.D., Cantab., F.R.S. Prorsssor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S. Proressorn WARRINGTON YORKE, M.D. Proressor B. BLACKLOCK, M.D. THE INCORPORATED LIVERPOOL SCHOOL OF TROPICAL MEDICINE FouUNDED BY SiR ALFRED LEWIS JONES, K.C.M.G (A filiated with the University of Liverpool) Hon. President: H.R.H. PRINCESS CHRISTIAN (Princess of Great Britain and Ireland) Chairman: Str FRANCIS C. DANSON Vice-Chairman: Professor R. CATON, C.B.E. Hon. Vice-Presidents: VHE EarL or Dersy, K.G. VISCOUNT MILNER, G.C.B. VISCOUNT Prrriz, K.P. VISCOUNT LEVERHULME Sir Epwarp MEREWETHER, K.C.V.O. S1R OWEN Puivipps, K.C.M.G. Mr. O. HARRISON WILLIAMS COMMITTEE Sir H. J. Reap, K.C.M.G. Colonial Office Vice-Chancellor J. G. Apamt, F.R:S. University of Liverpool Professor R. CATON, C.B.E. Mr. H. WAvDE DEACON | Council of University of Liverpool Professor J. M. Beattie, M.D. ] b ; Se Professor W. J. DaKIN, D.Sc. : Senate of University of Liverpool Mr. E.G. BuckLEY Mr. T. WooDSEND Mr. T. F. HARRISON } Mr. ENFIELD E. FLETCHER Royal Southern Hospital Steamship Owners’ Association Mr. W. J. B. CHAMBERS ) Oe ak XS a. Vin ee j Shipowners’ Association Mr. R. B. MILLER West African Trade Association Mr. G. BROCKLEHURST Mr. H. D. DickIE Professor E. GLYNN Professor Sir W. HERDMAN, C.B.E., F.R.S. Professor E. W. Hope, O.B.E. Mr. DAvip JONES Mr. J. PICKERING JONES Captain R. RANKIN Professor J. W. W. STEPHENS, M.D., F.R-.S. Professor R. NEWSTEAD, M.Sc., F.R.S. Professor W. YORKE, M.D. Professor B. BLAcKLock, M.D. Mr. J. A. TINNE, Hon. Treasurer Mr. J. L. McCarrny, Secretary J. Mipptemass Hunt, H 24+-25, Exchange Buildings, Liverpool Hon. Dean Staff, 1922 Alfred Fones Professor of JOHN WILLIAM WATSON STEPHENS, M.D., Cantab., Tropical Medicine . : . F.R.S. . u Dutton Memorial eae of Entomology . . ROBERT NEWSTEAD, J.P., F.R.S., M.Sc., A.L.S., F.E.S. Walter Myers eal of Parasitology . = . WARRINGTON YORKE, M.D. Professor of shea Diseases Africa . . BREADALBANE BLACKLOCK,*M.D. Lecturer on Entomology ° . ALWEN M. EVANS, M.Sc. Assistant Lecturer on Entomology . W. H. POTTS, B.A. Lecturer on Protozoology . PHILIP A. MAPLESTONE, D.S.O., M.B., Ch.B. Lecturer on Helminthology . . T. SOUTHWELL, M.Sc., A.R.C.Sc. Hon. Lecturer on Clinical Veterinary Parasitology . . A. W. NOEL PILLERS, F.R.C.V.S. Assistant Lecturer and Demonstrator on Veterinary Parasitology . Vacant. Lecturer on Tropical Surgery . ROBERT ERNEST KELLY, C.B., M.D., F.R.C.S. Lecturer on Tropical Sanitation . R. H. KENNAN, M.D. Hon. Lecturer on Tropical Sanitation WILLIAM THOMAS PROUT, M.B., C.M.G. Hon. Statistician ©. . . WALTER STOTT. Royal Infirmary, Liverpool Physician. . . +. +. JOHN WILLIAM WATSON STEPHENS, M.D,, Cantab., . ERS. Assistant Physician . P . WARRINGTON YORKE, M.D. House Physician and Clinical Pathologist : - RUPERT MONTGOMERY GORDON, M.D.,-Ch.B. Consulting Surgeon. 3 . W. THELWALL THOMAS, F.R.C.S. The Manaos Research Laboratory Director . : . A » HAROLD WOLFERSTAN THOMAS, M.D., C.M. Sierra Leone Research Laboratory Director . . . +. ~~. BREADALBANE BLACKLOCK, M.D. Research Assistants . - . S. ADLER, M.B., Ch.B. EDWARD J. CLARK, M.B. THE MARY KINGSLEY MEDAL This medal was struck in commemoration of the work of the late Miss Mary Kingsley in West Africa, and is conferred in recognition of distinguished scientific achievement. HONORARY RECIPIENTS Her Royal Highness Princess Christian Lord Lister The Right Hon. Joseph Chamberlain Prince Auguste d’Arenberg Mrs. Pinnock Mr. William Adamson Professor William Carter RECIPIENTS 1905— Colonel Sir David Bruce, K.C.B. Geheimrath Professor Robert Koch Dr, A. Laveran Sir Patrick Manson, K C.M.G., 1907— Professor Danielewsky Dr. Charles Finlay Mr. W. M. Haffkine Professor Golgi Colonel Gorgas Professor Theobald Smith 1910— Sir William Macgregor, G.C.M.G. Professor R. Blanchard Dr, Anton Breinl Professor Angelo Celli Dr. C. W, Daniels Surgeon-General Sir Alfred Keogh Colonel W. G. King Professor Nocht Professor G. H. F. Nuttall Major Leonard Rogers Professor J. L. Todd Surgeon-General Walter Wyman 1913— Professor Fred V. ‘Theobald 1917— Dr. Griffith Evans 1919— Dr. J. W. Scott Macfie The Oswaldo Cruz Institute, Rio de Janeiro b 1920— Major E, E. Austen, D.S.O. Dr. A. G, Bagshawe, C.M.G. Dr. Andrew Balfour, C.B. Dr. A. L. G. Broden Mrs. Chalmers, in recognition of the work of the late Dr. A. J. Chalmers Professor B, Grassi Professor R. T. Leiper Professor F. Mesnil Dr. Edmond Sergent Dr. C. W. Stiles Dr. T. Zammit THE ALAN H. MILNE MEDAL This medal was struck to commemorate the late Alan H. Milne, C.M.G., the first Honorary Secretary of the School (1899-1917), and is awarded twice yearly on the recommendation of the examiners for the Diploma in Tropical Medicine. 1921— George Phillip Parmer Allen 1922— Quinton Stewart NOTICE The following courses of instruction are given by the Liverpool School of Tropical Medicine each year :— (1) Two courses for the Diploma in Tropical Medicine, each of three months’ duration, commencing about the 15th September and the 7th January. The D.T.M. examinations are held in December and April. (2) A short advanced course, of one month’s duration, in June. (3) Two courses in Veterinary Parasitology, each of three months’ duration, commencing about the 15th September and the 7th January. DIPLOMA IN TROPICAL MEDICINE The Diploma shall be awarded only to candidates who possess a qualification to practice Medicine recognised for this purpose by the University, and who present satisfactory certificates of having attended approved courses of study, and pass the prescribed examination. FEES Diploma Course _... iF ae as Twenty Guineas Short, Advanced Course ... ee Six Guineas Course in Veterinary Parasitology — oe Fifteen Guineas Diploma Examination BS : 203 Five Guineas Fee for the use of a School microscope deine one term ... One Guinea For prospectus and further information, application should be made to the Hon. Dean, School of Tropical Medicine, University of Liverpool. The following have obtained the Diploma in Tropical Medicine of the University of Liverpool :— Diploma in Tropical Medicine Date of Date of Diploma Diploma 1904 Augustine, Henry Joshua 1905 Macfarlane, Robert Maxwell 1904 Bennett, Arthur King 1g05 Maddock, Edward Cecil Gordon 1904 Bruce, William James 1g05 Moore, James Jackson 1904 Byrne, John Scott 1g05 Nightingale, Samuel Shore 1904 Clayton, Thomas Morrison 1905 Radcliffe, Percy Alexander Hurst 1904 Dalziel, John McEwen 1905 Young, John Cameron 1904 Dee, Peter 1904 Greenidge, Oliver Campbell 1906 Adie, Joseph Rosamond 1904 Hehir, Patrick 1906 Arnold, Frank Arthur 1904 Khan, Saiduzzafor 1906 Bate, John Brabant 1904 Laurie, Robert 1906 Bennetts, Harold Graves 1904 Maclurkin, Alfred Robert 1906 Carter, Robert Markham 1904 McConnell, Robert Ernest 1906 Chisholm, James Alexander 1g04 Nicholson, James Edward 1906 Clements, Robert William 1904 Philipson,” Nicholas 1906 Dundas, James 1904 Sharman, Eric Harding 1906. Faichnie, Norman 1904 Thomson, Frank Wyville 1906 Jeffreys, Herbert Castelman 1904 Walker, George Francis Clegg 1906 Mackenzie, Donald Francis 1906 Pailthorpe, Mary Elizabeth 1g05 Anderson, Catherine Elmslie 1906 Palmer, Harold Thornbury 1905 Brown, Alexander 1906 Pearse, Albert 1905 Caldwell, Thomas Cathcart 1906 Sampey, Alexander William 1905 Critien, Attilio 1906 Smithson, Arthur Ernest 1905 Hooton, Alfred 1906 Taylor, Joseph van Someron 1905 Hudson, Charles Tilson 1906 Taylor, William Irwin 1g05 Illington, Edmund Moritz 1906 Tynan, Edward Joseph Date of Diploma 1906 1906 1906 1907, ez 1907 (907 Abe// 1907 eey/ Ler ey/ 1907 907, L2e7/ BOOT, ushey) 1907 {3 27/ ROOF, 1907 1907; 1907) 27) 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1908 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909 1909) #909 Watson, Cecil Francis Willcocks, Roger Durant Williamson, George Alexander Allan, Alexander Smith Allwood, James Aldred Bond, Ashton Branch, Stanley Collinson, Walter Julius Davey, John Bernard Donaldson, Anson Scott Fell, Matthew Henry Gregson Gann, Thomas William Francis Graham, James Drummond Hiscock, Robert Carroll Keane, Joseph Gerald Kennan, Richard Henry Kenrick, William Hamilton Le Fanu, George Ernest Hugh Mackey, Charles Maddox, Ralph Henry McCarthy, John McDonald Raikes, Cuthbert Taunton Ryan, Joseph Charles Vallance, Hugh Caverhill, Austin Mack Crawford, Gilbert Stewart Dalal, Kaikhusroo Rustomji Dansey-Browning, George Davidson, James Dickson, John Rhodes Dowdall, Arthur Melville Glover, Henry Joseph Greaves, Francis Wood Goodbody, Cecil Maurice Harrison, James Herbert Hugh Joshi, Lemuel Lucas Le Fanu, Cecil Vivian Luethgen, Carl Wilhelm Ludwig Mama, Jamshed Byramji McCay, Frederick William McLellan, Samuel Wilson Pearce, Charles Ross Schoorel, Alexander Frederik Smith, John Macgregor Stewart, George Edward Tate, Gerald William Whyte, Robert Abercrombie, Rudolph George Allin, John Richard Percy Armstrong, Edward Randolph Barrow, Harold Percy Waller Beatty, Guy Carr-White, Percy Chevallier, Claude Lionel Clark, William Scott Cope, Ricardo Fleming, William Hanschell, Hother McCormick Hayward, William Davey Henry, Sydney Alexander Innes, Francis Alexander Jackson, Arthur Frame Kaka, Sorabji Manekji McCabe-Dallas, Alfred Alexander Donald Meldrum, William Percy Murphy, John Cullinan Date of Diploma 1909 1909 1909 1909 1909 Tg909 Ig10 19to 1910 Ig10 IgIo 1gIo 1910 1gI0 1gI0 IgIo 1g10 Ig10 1gI0 I1gIo 1g10 1gto 1910 1910 1910 gto 1gI0 Ig10 1910 IgIo 1910 IgI0 glo 1gIo Ig! IgII IgIt IgIt gtr git IgII IgII Igi! Igtt Igit IgII IgII IQIt IgII IgII IgII IgIt git Igtt IgIt 1912 1912 1gI2 I1g12 1912 1912 1912 1912 1912 1912 1912 1912 Samuel, Mysore Gnananandaraju Shroff, Kawasjee Byramjee Thornely, Michael Harris Turkhud, Violet Ackroyd Webb, William Spinks Yen, Fu-Chun Brabazon, Edward Castellino, Louis Caulcrick, James Akilade Dowden, Richard Haigh, William Edwin Hamilton, Henry Fleming Hefferman, William St. Michael Hipwell, Abraham Homer, Jonathan Houston, William Mitchell James, William Robert Wallace Johnstone, David Patrick Korke, Vishnu Tatyaji Macdonald, Angus Graham Macfie, John Wm. Scott Manuk, Mack Walter Murison, Cecil Charles Nanavati, Kishavlal Balabhai Nauss, Ralph Welty Oakley, Philip Douglas Pratt, Ishmael Charles Sabastian, Thiruchelvam Shaw, Hugh Thomas Sieger, Edward Louis Sousa, Pascal John de Souza, Antonio Bernardo de Waterhouse, John Howard White, Maurice Forbes Blacklock, Breadalbane Brown, Frederick Forrest Chand, Diwan Jai Holmes, John Morgan Tevers, Charles Langley Iles, Charles Cochrane Ingram, Alexander Kirkwood, Thomas Knowles, Benjamin Liddle, George Marcus Berkeley Lomas, Emanuel Kenworthy Mackarell, William Wright MacKnight, Dundas Simpson Mascarenhas, Joseph Victor Murray, Ronald Roderick Oluwole, Akidiya Ladapo Rao, Koka Ahobala Sinton, John Alexander Tarapurvalla, Byramji Shavakshah Taylor, John Archibald Woods, William Medlicott Aeria, Joseph Reginald * Anderson, Edmund Litchfield Borle, James Bowie, John Tait Brassey, Laurence Percival Christie, David Dillon, Henry de Courcy Dunn, Lillie Eleanor Hardwicke, Charles Jagose, Jamshed Rustomji Kochhar, Mela Ram : McGusty, Victor William Tighe Date of Diploma giz giz 1912 1912 1g12 I1g12 1gI2 Ig12 1gI2 1gI2 1g12 1912 1gt2 Igt2 1913 1913 1913 1913 £OX3 1913 1913 1913 1913 T9X3 1913 1913 1913 IQX3 T1913 1913 1913 1913 1913 1913 1913 T1913 1913 1913 1913 1913 1913 1913 1913 1913 1914 1gt4 1914 1914 1gt4 1914 1gi4 1914 1914 1914 1gt4 1914 1914 1gt4 1914 1914 1gi4 1914 1914 1914 1915 Tors 1915 1915 1915 Milne, Arthur James Mitra, Manmatha Nath Myles, Charles Duncan Pelly, Huntly Nevins Prasad, Bindeshwari Prentice, George Ross, Frank Russell, Alexander James Hutchison Ruthven, Morton Wood Sandilands, John Seddon, Harold Smalley, James Strickland, Percy Charles Hutchison Watson, William Russel Austin, Charles Miller Banker, Shiavux Sorabji Becker, Johann Gerhardus Carrasco, Milton Clark, James McKillican Forsyth, Charles Grahame, Malcolm Claude Russell Grieve, Kelburne King Hargreaves, Alfred Ridley Hepper, Evelyn Charles Hiranand, Pandit Jackson, Oswald Egbert Khaw, Ignatius Oo Kek MacKelvie, Maxwell MacKinnon, John MacPhail Macmillan, Robert James Alan Mouat-Biggs, Charles Edward Forbes Noronha, John Carmel O'Connor, Edward Olubomi-Beckley, Emanuel Pestonji, Ardeshir Behramshah Puttanna, Dodballapur Sivappa Reford, John Hope Smith, Edward Arthur Stewart, Samuel Dudley Walker, Frederick Dearden Wilbe, Ernest Edward Wilson, Hubert Francie Yin, Ulg Ba Young, William Alexander Arculli, Hassan el Chohan, Noormahomed Kasembha Connell, Harry Bertram Gerrard, Herbert Shaw Gimi, Hirji Dorabji Gwynne, Joseph Robert z Hodkinson, Samuel Paterson Jackson, Arthur Ivan Kaushash, Ram Chander Kelsall, Charles Luanco y Cuenca, Maximino Misbah, Abdul-Ghani Naguib Naidu, Bangalore Pasupulati Balakrishna Rowe, John Joseph Stephen Roy, Raghu Nath Shiveshwarkar .Ramchandra Vishnu Sur, Sachindra Nath Talati, Dadabhai Cursedji Wilkinson, Arthur Geden Wright, Ernest Jenner Lobo, John Francis Madhok, Gopal Dass Pearson, George Howorth Swami, Karumuri Virabhadra Wood, John Date of Diploma 1916 1916 1916 1916 1916 1916 1916 EOL, 1917 1917 1918 wy Behe) 1929 1919 1919 Re) 19'9 OM) oe 1gIg 1919 IgIg pote ee) 1919 T1919 1920 1920 1g20 1920 1920 1920 1920 1929 1922 1920 1920 1920 1920 1920 1921 1921 1921 1921 1921 1921 1921 1921 1921 1g2t 1921 1921 1921 1922 1922 1922 1922 1922 1922 1922 1922 1922 1922 1922 1922 Barseghian, Mesroob Chaliha, Lakshmi Prasad Lim, Albert Liat Juay Lim, Harold Liat Hin Metzger, George Nathaniel Soderstrom, Erik Daniel Wheeler, Louis Chapman, Herbert Owen Krishnamoorthy, Yedatore Venkoba Lipkin, Isaac Jacob Watts, Rattan Claud Bowle-Evans, Charles Harford Burnie, Robert McColl Celestin, Louis Abel Cummings, Eustace Henry Taylor Darling, Georgina Renington Drake, Joan Margaret Fraser Fraser, William James Gordon, Rupert Montgomery Krige, Christian Irederick Maplestone, Philip Alan Oluwole, Isaac Ladipo Rustomjee, Khusshuyee Jamesidjee Sawers, William Campbell Thompson, Mary Georgina Turner, Gladys Maude Young, Charles James Adler, Saul Anderson, William Jenkins Webb Campbell, George Cobb, Charles Eric Cobb, Enid Margaret Mary Connolly, Evelyn Mary Fernandez, Daniel David Lim. Chong Eang McHutcheson, George Browne van der Merwe, Frederick O'Farrell, Patrick Theodore Joseph Renner, Edowo Awunor Vaughan, James Churchwill Waller, Harold William Leslie Allen, George Phillip Farmer Corfield, Charles Russell Hamid, Abdul Longhurst, Bell Wilmott Macvae, George Anthony Madan, Hans Raj Mulligan, William Percival Nixon, Robert Richmond, Arthur Stanley Shri Kent, Shamsher Singh Skinner, James Macgregor Stewart, Robert Bell Thomson, Marion Bhatia, Jagat Ram Cohen, Morris Joshua Crawford, Andrew Clemmey Gilmore, Edward Raymond Gracias, Cajetan Manuel Jennings, Arthur Richard Lethem, William Ashley Paul, Sachchidananda Hoshen Pinder, John Rieley, Stanley Desmond Rutherford, Gladys Stewart, Quinton ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY EDITORIAL NOTICE Articles for publication should not exceed twenty-five pages of the Annals, and will be understood to be offered alone to this Journal. They should be typewritten and addressed to:—The Editors, School of Tropical Medicine, The University, Liverpool. Illustrations for text figures or charts should be drawn clearly and firmly in Indian ink, if possible on Bristol board. N.B.—Blwue or other coloured ruling in squares or lines cannot be reproduced. All lettering, names or legends on text-figures, charts or maps should be printed sujffczently large to allow of clear legibility on reduction if necessary. Plates and illustrations should be accompanied by short explana- tions. References to authors in the text must be made in the following way :—‘ According to Smith (1900) the spleen is enlarged, but Robinson (1914) says the reverse.’ The references should be collected in alphabetical order of authors’ surnames at the end of the paper, and arranged in the following way :— ROBINSON, S. (1914). ‘The spleen in malaria.’ Annals of Nosology, Vol. XX, pp. 20-25. Sir, J. (1900). ‘Enlargement of the spleen in malaria.’ Yournal of Pathometry, Vol. I, pp. 1-20. Twenty-five reprints are supplied of each paper, free of charge. Additional copies (up to 200) can be supplied at cost price. Subscription: {1 2s. 6d. per volume, post free, payable in advance to the Secretary, The University Press of Liverpool Limited, 177 Brownlow Hill, Liverpool, to whom correspondence concerning advertisements should also be addressed. 341 AN UNUSUAL TYPE OF NODULAR LEPROSY IN THE SUDAN BY Ro GaARGHIBALD aDiS Oo. MD. (DIRECTOR, WELLCOME TROPICAL RESEARCH LABORATORIES, KHARTOUM) (Received for publication 20 August, 1922) PLATE XV Leprosy has a wide distribution in the Sudan, but is by no means the common entity among the native population as in the countries of the Far East. Of the varieties observed, the nodular or tubercular type is apparently the more common, presenting little difficulty in clinical diagnosis, and occurring usually in the form of well-developed nodular or tubercular lesions of the skin and tissues, as illustrated in a recent paper published by the writer. The case, which forms the subject of this paper, differed clinically from the usual type of leprosy observed in the Sudan, and appears worthy of record, inasmuch as it presented certain features which certainly obscured the diagnosis. The patient was an Egyptian, about 30 years of age, who stated he had suffered from an eruption of the skin for a period of one year. According to his history, the eruption apparently commenced on the face in the form of small shotty papules, similar ones eventually appearing on the forehead, ears, trunk and upper and lower extremities. The eruption caused little or no inconvenience, but as it appeared to be getting more extensive and causing some disfigurement, he sought medical advice. His previous medical history contained little of interest. There was no history of syphilis; the patient, however, admitted that his wife had an abortion a few months previously. The case having presented certain clinical features akin to a syphiloderma, and as facilities for proving this by laboratory examination were lacking, he was treated 342 with a course of injections of ‘606,’ but failed to show any improvement; in fact, his condition became progressively worse. When seen by the writer, the patient was well nourished and in fair general health. On examination, it was found that the skin of the face, neck, anterior and dorsal aspects of the trunk, and the. flexor and extensor aspects of the arms and legs showed numerous miliary papules varying from 03 to 0'5 centimetres in diameter. The majority of these were discrete, with a smooth surface, circular contour, pink colour, and of a shotty consistency; some of them showed a slight inflammatory reaction at the base. In certain areas, more especially on the neck and arms, many of the papules showed a circular depression or umbilication in the centre, while others showed simply a pale central area (Plate XV, fig. 1). No pustula- tion was noted. The largest were on the face, and here the majority of them were discrete, whereas those on the ears had coalesced and caused considerable thickening of the tissues, producing an appearance not unlike that of haematoma auris. Papules were also present over both superciliary regions, where a slight degree of madarosis was noted. The skin of the arms was more affected than that of the lower extremities, both flexor and extensor aspects being involved. The intervening portions of the skin presented no abnormalities, except in a few areas on the face where there was a certain degree of erythema. No nodules or ulcers were detected in the buccal mucous membrane; but the posterior fauces and larynx were slightly inflamed, which accounted for the somewhat hoarse voice of the patient. The submaxillary and axillary lymphatic glands were slightly enlarged and firm on palpation. No abnormalities were detected in the heart, lungs, liver and spleen. The patient’s temperature at the time of examination was normal, but he admitted that he suffered occasionally from attacks of pyrexia. Two of the shotty papules from the arm were excised, fixed in picric alcohol, and embedded for sections. Microscopical examina- tion of haematoxylin-eosin stained preparations showed the cytological changes associated with a granuloma, and _ special staining methods demonstrated the presence in the tissues of large numbers of acid-fast bacilli, morphologically resembling leprosy bacilli 343 (Plate XV, fig. 4). These were especially well seen in sections stained by carbol-fuchsin, decolourized in 10 per cent. sodium sulphite, and finally counterstained with an aqueous solution of methylene blue containing I per cent. sodium carbonate. Histopathology of a nodule Sections showed a thinning not only of the horny layer of the epidermis, but also of the rete mucosum, the cells of the latter consisting chiefly of oval and columnar cells (Plate XV, figs. 2 and 3). Beneath the lower border of the rete mucosum there was a marrow zone, poor in cellular elements, which stained feebly with tissue stains (fig. 3). Special staining reagents showed it was composed of fibrous tissue, which apparently had undergone a hyaline or vitreous degeneration. Beneath this narrow zone there was a marked cellular reaction in the upper part of the corium. The cells here were composed chiefly of plasma and lymphoidal cells (fig. 2); but no giant cells were present. This cellular infiltration occurred also to a less degree in the pars reticularis, but it varied in intensity in different areas of the nodules. Where well marked, it encroached on the narrow or vitreous zone, extending almost to the rete mucosum (fig. 2). Where it was slight or hardly present the vitreous zone was wider, and beneath it the corium appeared to be composed of loose, oedematous-looking connective tissue in which dilated lymphatic vessels filled with lepra bacilli were noted (fig. 4). This area was rich in lepra bacilli, dense masses extending throughout the pars reticularis down to the subcutaneous tissues ; they were not found either in the sebaceous glands or in the hair follicles; indeed, these structures, like the blood vessels, appeared to be unaffected. In the vitreous zone, beneath the rete mucosum, only a few single bacilli were found; none were located in the rete or in the horny layer. The infected area appeared to be confined to the corium, the infection reaching that portion of the skin via the lymphatics. Remarks on the Case In considering the condition from a clinical aspect, it must be admitted that the case presented certain puzzling features. The discrete nature of the eruption and the umbilication of some 344 of the nodules, their size and extensive distribution, together with the clinical history of only twelve months’ duration, compelled one to consider and eliminate various skin eruptions that have been studied in this country. Of these may be mentioned Molluscum contagiosum, generalised vaccinia, Lichen hyperkeratosis, cutaneous Leishmaniases, prurigo, yaws and syphiloderma. Most of these could be readily differentiated ; the possibility of the case being one of leprosy, occurring, moreover, in a Government official, did not occur to the writer, nor was it suspected by the various medical men who examined the case. It was left to the histological examination of the excised nodules to throw light on the nature of a condition which might well be termed miliary leprosy. yr a. te att “Aleriir, ‘ im Hees (} eget 1 ‘ "ray : “ly Tia glib atiery ma ineetow Lory p te eb; +2 ‘ee = to os Tyr ar . : aye hoes a) A anos sitileved GR )”. ee . ’ wird " bestvsl - was i} inehsatey Mite - ‘. ¥ a vs Ja She ORE Mw Doriists mititay Sit) to nicntsce & we dqstgorheios 28 a ainie dish SdT © ould snsbedrsin nisdisyi-| ot » iit siqol do qmala jaoesty Fig. Fig. Fig. lee 346 EXPLANATION OF PLATE XV Illustrating the eruption. Photo-micrograph of a section of a papule, showing the cellular infiltration encroaching on the rete malpighii. x 190. Photo-micrograph of a section of a papule, showing the narrow hyaline zone of degeneration subjacent to the rete malpighil. x 190. Photo-micrograph of a section of the corium stained with carbol-fuchsin methylene blue. The dark stained areas represent clumps of lepra bacilli. x 800. Annals Trop. Med, & Parasitol., Vol, XVI PLATE XV C. Tinling & Co., Litd., Imp. 347 ANCYLOSTOMA BRAZILIENSE BY CLAYTON LANE (Received for publication 2 September, 1922) In a recent paper, Dr. Gordon (1922) reports finding Ancylostoma braziliense in man in four out of sixty-four autopsies performed in Manaos, Amazonas, Brazil, and concludes thus :— “The comparison of these worms and other two-toothed ancylo- stomes from dogs and cats in North Brazil and India, and also from cats in South Africa and dogs in West Africa, failed to show the difference claimed to exist by de Faria between A. ceylanicum and A. braziliense.’ It is very desirable that a decision on the matter of identity of these worms should be generally accepted, and the first step necessary seems to be a historical survey rather fuller than that which Gordon supplies. Gomes de Faria (1910) described Axcylostoma braziliense from Felis domesticus and Canis familiaris in Brazil. Looss (1911) described Axncylostoma ceylanicum from the civet cat, Vzverricula malacensis, in Colombo, Ceylon. Leiper (1913), without examina-~ tion of A. braziliense, suggested, from the appearance of the dorsal! ray as figured by de Faria, that the two forms were identical, this ray having, he stated, a pair of digitations only on each of its two branches (Leiper (1915)), a statement which, however, requires alteration (Clayton Lane (1916)). Clayton Lane (1913) first recorded A. ceylanicum as a parasite of man, a fact since amply confirmed from various parts of the world, thereby giving to the question of nomenclature a medical interest. De Faria (1914) published a short paper in which he quotes a letter from Looss, who therein states emphatically that A. braziliense has only a single tooth on each ventral tooth plate; that its bursal rays, especially the externo-dorsal, are remarkable for their length and delicacy; and that the relative thickness of the bursal rays is a definite differential [specific] character, De Faria (1916), after examining abundant 348 Brazilian material and comparing it with specimens of A. ceylanicum sent by Clayton Lane from Bengal, verifies the existence of the inner pair of teeth, which he describes as much smaller than are those of the Indian forms, but holds, nevertheless, that this comparative examination disposes completely of Leiper’s suggestion mentioned above. One of Looss’s specific criteria being thus swept away, the specific differences held to obtain between the two forms rested upon the relative slenderness of the bursal rays. In this relation, Gordon published measurements of the externo-dorsal ray of Brazilian forms and of forms supplied to him from Bengal by Clayton Lane. These measurements provided him with no constant differences, nor could he detect other constant distinctions between worms from these two areas or from Africa. The present intervention is prompted by two motives. The first is that the writer is credited by Gordon with supporting de Faria in his basis of specific differentiation. This is not exactly the case. What he actually did (Clayton Lane (1916)) was to comment upon the complete absence in existing descriptions of measurements of the internal organs; to express disbelief in Looss’s statement that only a single pair of teeth existed, it being inconceivable that de Faria should describe and draw a non-existent tooth; to accept Looss’s and de Faria’s statements that the bursal rays of the Brazilian form were strikingly fine; and to point out that, accepting this as a fact, there emerged the almost certain conclusion that two species were being dealt with. There was this significant addition, ‘It will probably be generally felt that there must be a thorough and independent examination by another experienced helminthologist before the question can be considered as settled.’ This examination has been made by Gordon, but even his published report leaves certain matters doubtful. The receipt of some material furnished by his courtesy, together with the importance of settling definitely, if possible, the specific name of a parasite of man (the second of the motives to which reference was made above) prompts the present note. An examination of the appended Table of Measurements mainly dealing with the internal organs of these forms, published apparently for the first time so far as the Brazilian ancylostomes are concerned, affords no justification for the duality of species. On 349 Fic. 1. Ancylostoma braziliense Brazilian forms : A, B, and C, the bursae of males. D, E, and F, the tooth-plates of males. Indian forms : G, the bursa of the male. H, I, and K, the tooth-plates; H and K of males, J of a female. Scales : L, scale for D, E, F, H, J, and K representing "1 mm. M, scale for 4, B, C, and G, representing ot mm. 35° the other hand, it gives no proof of unity of species, as is clear when one considers, for example, the relative measurements of the various members of the genus TZ7zchostrongylus. This question must under the circumstances be determined by shape, as, indeed, should always be the case. Taking first the male bursa: Figs. A, B, and C are from Gordon’s Brazilian material; fig. G from the dog in Bengal. Fig. B is typical of the stout-rayed condition generally ascribed to A. ceylanicum. In fig. A the rays are much finer. Fig. C shows a condition on the whole intermediate between the other two, although the externo-dorsal ray is short, ending far from the edge of the bursa, while the ventral rays and the internally-terminating lateral ones are pointed. This evidence demonstrates considerable individual variation upon those very points which are held constant within the species. This circumstance led naturally to a re-examina- tion of Indian material. Almost at once the form represented in fig. G was found. Its relatively fine lateral rays are not those associated with the accepted descriptions of A. ceylanicum, and yet they are from Indian material. Turning to the ventral oral plate, figs. D, E and F are from Gordon’s Brazilian material. The first shows a direct dorsal view from a male worm, with the inner, deeper teeth fairly marked. Fig. E is that of another male viewed dorso-laterally ; the obliquity brings into evidence and increases the apparent size of one deep tooth and obscures and minimises the other. In fig. F, clearing in creosote of this worm, which had lain long in lacto-phenol, was unsatisfactory and the deep teeth were invisible in a direct dorsal view. With lateral tilting their points could just be distinguished. Figs. B and F are from one and the same worm, apparently stout bursal rays being associated with apparent absence of the inner teeth. Figs. H, | and K are from Indian material, fig. I being from a female worm. Fig. K corresponds to fig. F. In it the inner tooth on one side is completely, and on the other almost completely, hidden by the large superficial outer tooth, and, had the specimen been imperfectly cleared, these would have been invisible. The evidence which has just been given shows that the bursal rays of these two-toothed forms, from whatever part of the world 351 they come, present great individual differences in length and width, the former partly real, partly apparent, and due to the fore- shortening caused by the incurving of the bursal edge; and that marked variations occur in the apparent size of the inner teeth, variations which can indeed, to some extent at least, be produced at will by the rolling of the worm. Both features are largely independent of the country of origin. Indeed, one must conclude that individual prepossession will play a preponderating part in determining whether any particular two-toothed ancylostome of this type is to be classified as A. draziliense or A. ceylanicum. In other words, there is no evidence offered that acceptable specific differences exist between individuals from the Old and New Worlds. In the absence of such evidence, Axcylostoma (Ceylancylostoma) ceylanicum (Looss (1911)) lapses as a synonym of Axcylostoma (Ceylancylostoma) braziliense (Gomes de Faria, (1910)). TABLE Measurements in millimetres of Males of Ancylostoma braxiliense from Brazil, and A. ceylanicum from Bengal. Brazilian Indian form form Oral cavity, length ... sth eae oe ae ies Ag O14 O'143 Oral cavity, transverse diameter... “3 bi: eas A 0°08 or0g Oral cavity, dorso-ventral diameter “ ate Ao 35 0°07 0°087 Nerve collar from head end ... ous wee tee a3 as o'7 0°57 Cervical papillae We or hee tb 80 ran ase O55 0°57 Excretory pore ” ox wee oe tee vee eee oss 0°57 Width oticuticular stration ec: 9 1s “ve son ses Tans 0°007 00075 Oesophagus, length ... ees Sie tee Sie ae a o'6 o'7 Oesophagus, breadth ... a wee fee Per sos ae ol O'ls GenwthiorMpiculesl cc say) 0) ges pecs) eel ach, | cave) ROB/tO O'9 o'8 Length of accessory piece... ote ars eae aes ay 0°065 0°75 Length of Cement gland... toe nee one neu. ueks 2'0 3°0 352 REFERENCES De Farta, Gomes (1910). Contribution towards the Classification of Brazilian Entozoa. Mem. Inst. Os. Cruz., Vol. I, No. 2, p. 286. — (1914). Ainda sobre o ‘ Agchylostoma braziliense’ (Gomes de Faria, 1910). Braz. Med., Vol. XXVIII, p. 113. (1916). Nota sobre Agchylostomea braxiliense G. de Faria, 1910. Mem. Inst. Os. Cruz., Vol. VIII, No. 2, p. 71. Gorvon, R. M. (1922). Ancylostomes recorded from sixty-seven Post-mortems performed in Amazonas. Ann. Trop. Med. & Parasit., Vol. XVI, p. 223. Lane, Crayton (1913). Agcebylostoma ceylanicum: A New Human Parasite. Indian Med. Gaz., Vol. XLVIII, p. 217. (1916). The Genus Ancylostoma in India and Ceylon. Indian fourn. Med. Research, Vol. IV, No. 1, p. 73. Lerrer, R. T. (1913). The apparent identity of Agchylostoma ceylanicum (Looss, 1911) and Agchylostoma braziliense (Gomes de Faria, 1910). Fourn. Trop. Med. & Hyg., Vol. XV1, P- 334+ (1915). Notes on the occurrence of Parasites presumably rare in Man. ‘Fourn. Roy. Army Med. Corps. Vol. XXIV, p. 569. Looss, A. (1gtr). The Anatomy and Life-history of Agchylostoma duodenale, Part 11. Records of the Egyptian Sckool of Medicine. 353 INTRA-UTERINE INFECTION WITH ANCYLOSTOMA CANINUM IN DOGS BY 5S, ADEER AND E. J. CLARK From the Sir Alfred Lewis Jones Research Laboratory, Freetown, Sierra Leone (Received for publication 3 September, 1922) Intra-uterine infection with hookworms has been noted by Howard (10917), who found ova in the stool of a child fourteen days old. Owing to lack of human material, we examined a number of young animals in order to find whether intra-uterine infection with hookworms is a common occurrence. Thirteen young dogs (from two to fifteen days old), representing eight different litters, were examined for ancylostomes. The results were as follows : — Age | A. caninum Litter | in days of | Number ee ee Remarks dogs examined | Worms Ova I 2 2 negative | negative 2 5 2 negative | negative : 3 5 2 positive negative | One infection was intense. Ancylos- (2) tomes up to 7 mm. long with well developed buccal capsules. No ova | in the uteri of the worms. 4 7 2 | negative | negative 5 13 I | positive | positive 6 I4 2 | positive positive (2) (2) 7 14 1 negative | negative 8 15 1 | positive | positive It thus appears that in Freetown, where intense infections with A. caninum are the rule in dogs, intra-uterine infection is common. 354 It is noteworthy that, although infection with A. ceylanicum 1s common in adult dogs, we have not found evidence of intra-uterine infection with this parasite. Infection of the foetus is possible in two ways :— (1) By larvae passing through the maternal blood stream to the placenta, and through the placenta to the foetus. (2) By larvae finding their way into the peritoneal cavity of the mother and passing through the uterine muscle to the placenta. Yoshida (1920) has shown the possibility of this by observing ancylostome larvae in the peritoneal cavity of experimentally infected guinea-pigs; and we have found ancylostome larvae in the peritoneal cavity of a guinea-pig which had been placed for ten hours in a vessel containing a mixed culture of A. caninum and A. ceylanicum. REFERENCES . Howarp, H. H. (1917). Pre-natal Hookworm Infection. Famaica Public Health Bul., pp: 20-24, Ext. Trop. Dis. Bull., March 15, 1920. Yosurpa, Sapao (1920). A New Course for Migrating Ancylostoma and Strongyloides Larvae after Oral Infection. ‘fourn. of Parasit., Vol. VII, No. 1, pp. 46-48. 355 CESTODES FROM INDIAN BIRDS WITH A NOTE ON LIGULA INTESTINALIS BY T. SOUTHWELL, M.Sc., A.R.C.Sc., F.Z.S. (Received for publication 25 September, 1922) A few species of cestodes dealt with below were presented to the author by Lt.-Col. Clayton Lane, I.M.S. The rest of the collection (except Ligula) were obtained from animals which died in the Zoological Gardens, Calcutta, on which post mortems were made in the Indian Museum. The following species are recorded in this paper :— PARASITE Tetrabothrius erostris Davainea tetragona ” ” a” ” Davainea (? tetragona) Davainea microscolecina Davainea polychalix Davainea cruciata Davainea sp. Davainea urogalli Davainea tragopani, n.sp. Davainea centropi, n.sp. Cotugnia fastigata Dilepis cypselina Dilepis campylancristrota Choanotaenia decacantha Choanotaenia (? octocantha) Choanotaenia microsoma, n.sp. Cyclorchida omalancristrota Rhabdometra tomica Hymenolepis medici ts Jusus ” ” * lanceolata Host Sterna bergit Pavo muticus Pavo cristatus Francolinus vulgaris Pavo nigropennis Eclectus vioratus Eos ricinata Lorius garrulus Pica rustica Crow pheasant Tragopan pheasant Tragopan pheasant Centropus rufipennis Ptistis coccineopterus Dendrocitta leucogaster Herodias garzetta Ardeola grayi Gallinago sp. Snipe Ploceus atrigula Melophus melanicterus Platalea sp. Francolinus pictus Pelicanus philippensis Larus brunneicephalus Hydropogne caspia Chenopis atrata 356 PARASITE Hymenolepis lanceolata ” ” 5, naja ” ” - zosteropis ” r» Hymenolepis farciminalis be stylosa re asymetrica . (? microcephala) “ (2 simplex) ih sp. ” ” ” annandalei usp. Echinocotyle uvalensis H ‘ymenolepis cap illaroides Diploposthe laevis ” ” 5 sp. (? laevis) Dioicocestus novae guineae Cestode sp. Ligula intestinalis Host Cygnus atratus Black swan Copschychus saularis Sitta chinensis Criniger flaveolus Melophus melanicterus Closa (?) chinensis Ploceus atrigula Dendrocitta sp. Pica rustica Brackypternus aurantius Trochalopterum meridionale Pica rustica Urocissa occipitalis Ciconia alba Tadorna cornuta Emberiza luteola Garrulax belangeri Oriolus melanocephalus Liothrix lutia Dendrocitta rufa Tadorna cornuta Limosa belgicae Snipe Gallinago sp. Snipe Netta rufina Nyroca ferina Strepsilas interpres Podiceps albipennis Sterna fluviatilus Danio acquipinnatus Family TETRABOTHRIIDAE, Ransom, 1909 Tetrabothrius erostris (Loennberg, 1889), Fithrmann, 1899 Three specimens without heads from intestine of Sterna bergit. Tamblegam, Ceylon, 6.9.12. the Indian Museum. Lake Numbered Z.E.V. Sony in the collection of / Family DAVAINEIDAE, Fihrmann, 1907 Sub-family Dav aingéINAr, Braun, 1900 Davainea tetragona (Molin, 1858), R. Blanchard, 1891 1. About fifty large specimens from intestine of Pavo muiticus. Zoological Gardens, Calcutta. Collected by the author, 10.12.14. 357 2. About forty specimens, same host and locality. Collected by the author, 3.1.17. 3 About ninety specimens, same host and locality. Collected by the author, 4.4.18. 4. About twenty specimens, same host and locality. Collected by the author, 12.7.18. 5. Several large and complete specimens from Pavo cristatus (common pea-fowl). Zoological Gardens, Calcutta. Collected by the author, 17.4.18. 6. Two specimens without heads from intestine of black shouldered pea-fowl. Zoological Gardens, Calcutta. No date. 7. Several specimens from intestine of Francolinus valent (black Francolin). Zoological Gardens, Calcutta. Collected by the author, 30.12.13. Twelve entire specimens were mounted, and a number of detached heads. In many heads all the hooks had been lost. In others only the hooks on the suckers were missing ; in still others some of the rostellar and sucker hooks were missing. Only in two or three heads were the hooks complete. In no case were the pores irregular, being invariably unilateral. Most of the strobilae were old and full of ripe eggs, but quite a number were ripe but not gravid. These measured from 5 mm. to 3 cms. in length. Davainea (? tetragona) A few fragments without head from intestine of Pavo nigropennis (black shouldered peacock). Collected by Lt.-Col. Clayton Lane, I.M.S., Berhampur, Bengal, 15.5.12. Davainea microscolecina, Fihrmann, 1908 I. Five specimens from intestine of Eclectus vioratus (parrot). Zoological Gardens, Calcutta. Collected by the author, 22.1.14. Previously recorded from Eclectus rosatus. 2. Two specimens from intestine of Eos ricinata. Zoological Gardens, Calcutta. Collected by the author, 6.7.15. Some of these specimens shewed a number of ripe segments strongly impregnated with lime. As a result they would not clear in clove oil, but after decalcifying in acid alcohol for several days they cleared readily. This phenomenon was often noted whilst working out the collection of Indian Cestoda. 358 Davainea polychalix, Kotlan, 1920 1. Four specimens from intestine of Lorius garrulus. Zoological Gardens, Calcutta. Collected by the author, 15.3.17. 2. Two specimens, same host and locality. Collected by the author, 3.3.17. Davainea cruciata (Rud. 1819), Fiihrmann, 1908 One specimen from intestine of Pica rustica (magpie). Zoological Gardens, Calcutta. No date. Davainea sp. A few fragments without heads from intestine of a crow pheasant. Zoological Gardens, Calcutta. Collected by the author, 22.4.15. Davainea urogalli (Modeer, 1790), R. Blanchard, 1891 One specimen and several fragments from intestine of a Tragopan pheasant. Zoological Gardens, Calcutta. Collected by the author, 27.2.15. In these specimens the head was about 380m broad ; its length could not be accurately determined because it passed into the neck, but it appeared to be at least 500”. The suckers have a diameter of about 1504 and are armed with about 17 rows of hooks. The rostellum has a diameter of about 50, and is armed with a double row, each hook measuring 7 or 8. About 50 were counted, but a number of hooks had clearly been lost. The total number is probably less than roo. The muscular system is feebly developed and consists of a few scattered longitudinal fibres, internal to which there occur a few transverse strands. The ventral excretory vessels on each side are very large, having a diameter of about 120”. They communicate with each other transversely by an equally large tube, in the posterior part of each segment. The dorsal vessel on each side is minute and has a diameter of Io only. The parenchyma throughout the worm is greatly developed, and it is very spongy owing to the occurrence of numerous small excretory cavities. The pores are unilateral and are situated in the anterior half of the segment. The testes number about 36-40 ; nine or ten are situated on the pore side and the rest posterior to the ovary, and aporal. Each testis has a diameter of about 55” when mature. In full development they 359 extend from the dorsal to the ventral surfaces and from the anterior to the posterior margins. They lie strictly within the water vessels. The cirrus pouch lies across the antero-lateral angle and extends to the water vessel. The vagina is posterior to the cirrus pouch. Both the genital canals run between the dorsal and ventral excretory vessels. Meggitt (1921) states that a number of eggs occur in each capsule, whilst Shipley (1909) states that the eggs lie singly in the parenchyma. In the Indian species the eggs at first occur in numbers in each capsule, but when fully developed each capsule contains only one onchosphere. Fihrmann states that the eggs lie within the two ventral water vessels. In our specimens, sections shewed that a single discontinuous layer of eggs was closely adherent to the lateral wall of each vessel, but they did not extend beyond that limit. DAVAINEA TRAGOPANTI, usp. Two specimens from intestine of a Tragopan pheasant. Zoological Gardens, Calcutta. Collected by the author, 27.2.15. EXTERNAL ANATOMY Only one of the specimens possessed a head. This worm measured 8-5 mm. in length, and its greatest breadth was 6004. It was composed of 27 or 28 segments; the last segment measured 825, in length and 6004 in breadth. The second specimen (without head) measured 7 mm. in length and its greatest breadth was 600. It contained 27 or 28 segments. Head. This was 1804 broad and about 1251long. Without destroying the head it was impossible to obtain accurate details relating to the hooks, but 23 hooks were counted in what appeared to be half the circumference of the rostellum. It seems, therefore, that the total number of hooks present was about 46. They did not appear to be ina double row. Their exact shape could not be made out, but they appeared to be typical. They measured roy in length. The suckers are armed, but all the hooks had been lost except in a portion of one sucker, where there appeared to be from 4 to 6 rows. The neck measured about 300” in length and was present in both specimens. 360 INTERNAL ANATOMY Owing to lack of material the nervous, muscular, and excretory systems were not investigated. Genitalia. Testes. There are 6 or 7 testes and they first appear in about Segment IV. When fully mature they measure about 7ou. Usually there are four situated aporally, one or two posterior to the ovary, and a single testis on the pore side, posterior to the internal extremity of the cirrus. Vas deferens. The cirrus pouch when fully developed extends half-way across the segment ; in 2 or 3 cases it extends a little more than half-way across. It has very thick (? muscular) walls. In Segment XVII it measures 250s¢ long and 110% broad. The cirrus is peculiar in being AM Biel 0. Vg. VS. V. Fic. 1. Davainea tragopani, n.sp. Ripe segments, mounted whole, showing genitalia. c.—cirrus ; ¢.p.—cirrus pouch; 0.—ovary; t.—testes; v.—vagina; v.d.—vas deferens; v.g.—vitelline gland; v.s.—receptaculum seminis. x 210. a greatly dilated organ densely covered with minute spines, and almost filling the cirrus pouch. The cirrus pouch persists to the last segment. The vas deferens is short and very slightly coiled. No seminal vesicle was observed (fig. 1). The genital pores are unilateral and are situated a little anterior to the middle point of the lateral margin of each segment. 361 Ovary. The ovary, which first appears in about Segment VIII, is definitely bilobed, each lobe being globular, and composed of a number of rounded acini. In full development each lobe measures about 70, in diameter. Receptaculum and vagina. From the pore the vagina pursues a direct course to a point between the two lobes of the ovary where it dilates into a receptaculum seminis. Vitelline gland. This lies posterior to the ovary and is a conspicuous organ. In full development its transverse and anterior diameters measure about 60, (fig. I). c. A.M.B cel Fic. 2. Davainea tragopani, usp. Gravid segment, mounted whole, showing uterus. ¢.—cirrus pouch ; wf.—uterus. X 50. Uterus. This first appears as a small cavity immediately anterior to, and between, the two lobes of the ovary. It enlarges and eventually single eggs become isolated in the parenchyma. In the last few segments no trace of the excretory vessels could be seen in either specimen ; it is, therefore, impossible to say definitely whether the eggs extend beyond them ornot. But as there was a definite area between the edge of the segment and the eggs, it would appear that the latter lie internal to the excretory vessel (fig. 2). Eggs. These have a diameter of about 54 and the onchosphere of about 25/. DIAGNOSIS The species is related to the proglottina type. The following table gives details of the various species described which resemble D. proglottina in being of small size, and at the same time serves to shew the points in which D. tragopani differs from'related species. I have, unfortunately, 362 been unable to procure Kowalewsky’s paper. Fihrmann recently (1919) discussed the relationship of the first four species indicated in the table, and it would appear almost certain that D. vavians, D. dubius and D. proglottina var. dublanensis are synonyms of D. proglottina. The principal points in which D. tragopani, n.sp., differs from them all are :—(I) size; (2) number of segments; and (3) the unilateral pores. The type specimen has been returned to the Indian Museum, Calcutta. Taste I. | No. of No. of | Size of Length | Breadth Segments | Hooks | Hooks Suckers | Pores Testes | | | | mm. Inm. D. proglottina I's O's 2-5 80-95 Ome I row | regularly 22 0n | armed alternate one side D. varians 18 ; 4-6 44-50 ? 4-5 rows | regularly more than | armed alternate fe} | | D. dubius Bs3 CHK || FAL) 2 rows |7'I-8"4j| 4-6 rows | alternate 12-15 | 50-60 armed | | D. dublanensts 4°0 ? | 6 | ? armed ? irregularly ? alternate D. tetraoensis ... 2°3 0°35 g-10 2rows | oft armed | a.ternate about 30 | 120-130 | with | several | | rings D. minuta io) 04 8 ? oft unarmed | alternate 10-12 D. paucisegmentata 570 o'7 5 ? ? unarmed | unilateral 40 D. himantopodis bo) ? 7-8 2 rows 7/4 armed, irregularly 4 50 no neck alternate D. tragopant, n.sp. 8:0 | 06 27 2 rows Tol 4 rows, unilateral 6 | 46 armed DAVAINEA CENTROPI, nsp. Three specimens and two fragments from intestine of Centropus vufipennis (the common Caccal), Lake Tamblegam, Ceylon, October 1911. Numbered Z.E.V. 6103 / in the collection of the Indian Museum. 363 EXTERNAL ANATOMY The specimens measured from 2:5 cms. to 3:5 cms. in length and had a maximum breadth of about 1-5 mm. Head. The head is prominent and presents a truncated appearance ; it measured about 300m broad. Its length could not be determined owing to the fact that it merges into a very short neck. The suckers have a diameter of about 3004 ; each sucker bears on its margin about 15 rows of hooks each measuring about 8. The rostellum is relatively small and is armed with about 300 hooks measuring from gy to 11m in length and arranged in a double row. Segments. The segments are very much broader than long, all except a few at the posterior extremity being quite shallow. Their lateral posterior margins are produced as shown in fig. 3. | The genital pores are irregularly alternate being situated, and directed, anteriorly. AM.B, cet. Vic. 3. Davainea centropi, n.sp. Outline of four segments. 35. INTERNAL ANATOMY Muscular system. This system is poorly developed ; the longitudinal fibres are relatively scanty and consist of small bundles somewhat widely separated ; the bundles decrease in size externally. The transverse fibres lie internal to the longitudinal muscles and are also very scanty. No oblique or dorso-ventral fibres were seen (fig. 4). 364 Nervous system. A small single nerve strand was to be seen lateral to the ventral water vessel on each side. On the pore side the nerve was ventral to the cirrus pouch and vagina. Excretory system. This consists of a single ventral vessel on each side ; on the pore side it lies ventral to the cirrus pouch (fig. 4). V. oO. Fic. 4. Davainea centropi, n.sp. Transverse section showing cirrus pouch, vas deferens, vagina, ovary and muscular system. c.p.—cirrus pouch; /.m.—longitudinal muscle; 0.—ovary; t.—testes; v.—vagina; v.d.—vas deferens; v.g.—vitelline gland; w.v.—water vessel. Xx 130. Genitalia. Testes. The testes are about forty in number; they lie dorsal and anterior on each side of the ovary and extend beyond the ventral excretory vessel. They are somewhat oval in shape and, when fully developed, measure about 85 by 55, Vas deferens. Yhe vas deferens is remarkable in being very long. It extends half-way across the segment and is thrown into a large number of loops which occupy almost the entire field between the internal extremity of the cirrus pouch and the poral wing of the ovary. No seminal vesicle was observed. The cirrus pouch varies in length, extending from about half to three-quarters the distance between the lateral margin and the ventral excretory vessel (fig. 4). Ovary. The ovary is a relatively large bi-lobed organ lying ventral 365 and posterior; in full development it extends almost to the dorsal transverse muscle fibres (figs. 4 and 5). Fic. 5. Davainea centropi, n.sp. Transverse section showing male and female genitalia. X 70. Receptaculum and vagina. From the pore, the vagina runs dorsal to the cirrus pouch ; at the internal extremity of the latter organ, the vagina curves gradually and runs directly to the ovary. It is muscular throughout its length. Its internal extremity is dilated into a muscular receptaculum seminis, which, in full development, measures about 150, in length and 50 in breadth (fig. 4). The oviduct, vitelline duct, and fertilisation canal are noticeable on account of their length. Vitelline gland. This lies ventral to and between the two lobes of the ovary ; it is large and easily seen (figs. 4 and 5)- Uterus. In full development, the uterus extends beyond the ventral excretory vessels and consists of a large number of parenchymatous capsules, each containing a single onchosphere. Eggs. These havea diameter of about 55 ; the onchosphere measures about 36. DIAGNOSIS Up to the present only about fourteen species of Davainea have been recorded which have armed suckers, and irregularly alternating genital pores. The species just described differs very definitely from them all. I therefore consider the species new and have named it D. centropi. 366 Cotugnia fastigala, Meggitt, 1920 Three specimens from intestine of Ptistes coccineoplerus, Gould, 1865. Zoological Gardens, Calcutta. Collected by the author, 13.11.15. The specimens had the following measurements :— Taste II. = —— = | I 2 3 Length mae oo Be. oes oe A. 750mm. | 7o'O mm. 60°o mm. Greatest breadth ... Bee wk = 3°5 mm. 38 mm. 3°73 mm. Number of segments aa0 385 os 212 210 205 As Meggitt was unable to isolate and figure a complete rostellar hook, a drawing of a hook from the Indian example is given below (fig. 6). A.M.B., cel. Fic. 6. Cotugnia fastigata, Meggitt. Diagram of a hook. In our specimens the vagina was almost invariably situated some distance posterior to the cirrus pouch, and as a result it was impossible to determine whether the vagina was dorsal or ventral to the pouch. In 6 or 7 segments examined, however, the vagina was definitely dorsal to the pouch on one side and ventral on the other—a character peculiar to the genus Montezia. Family HYMENOLEPIDIDAE, Railliet and Henry, 1909 Sub-family Dipriipiin 4k, Stiles, 1896 Dilepis cypselina, Neslobinsky, Ig11 One fragment with a head, of what is almost certainly this species, was obtained from the intestine of Dendrocitta leucogaster (tree-pie) ; Zoological Gardens, Calcutta. Collected by the author, 7.12.15. 367 The head was armed with a double crown of about 90 hooks, measuring about 244. The genital pores were unilateral. The cirrus pouch was situated anteriorly, and extended almost to the water vessel. Dilepis campylancristrota (Wedl, 1855), Fihrmann, 1908 1. Four specimens from intestine of Herodias garzetta (paddy bird), Berhampore, Bengal. Collected by Lt.-Col. Clayton Lane, I.M.S., June, Ig12. Numbered Z.E.V. sr in the collection of the Indian Museum. 2. Numerous specimens from Ardeola grayi (pond heron), Zoological Gardens, Calcutta. Collected by the author, 14.12.13, and numbered Z.E.V. a in the collection of the Indian Museum. / Choanotaenia decacantha, Fihrmann, 1913 Four specimens from intestine of a snipe (Gallinago sp.), Berhampur, Bengal. Collected by Lt.-Col. Clayton Lane, I.M.S., 17.12.12. The specimens agreed with Fiihrmann’s description except in the following minor details :— (z.) The hooks measured 23-4; in the type specimen they measured 19:8 to 21-6p. (2.) The type specimen had from 40 to 50 segments; the Indian forms have from 40 to 98 segments. Choanotaenia (? octocantha, Fiihrmann) I. One specimen,, without head, from intestine of a snipe, Berhampur, Bengal. Collected by Lt.-Col. Clayton Lane, I.M.S., 12.3.12. 2. One specimen from same host and locality. Collected by Lt.-Col. Clayton Lane, I.M.S. (219 b), 17.12.12. CHOANOTAENIA MICROSOMA, usp. I. Six specimens from intestine of Ploceuws atrigula (the eastern baya). Zoological Gardens, Calcutta. Collected by the author, 26 10.15. 2. About twelve specimens from intestine of Melophus melanicterus (the crested bunting). Zoological Gardens, Calcutta. Collected by the author, 25.6.15 368 EXTERNAL ANATOMY The worms measure from 4 mm. to 8 mm. in length and have a maximum breadth of about 630". They consist of from 25 to about 50 segments. Head. The head is square and measures about 220”; the suckers have a diameter of about 140. The rostellum measures about 180, in length and has a diameter of about 50. Its anterior extremity is expanded and has a breadth of about gow and a length of 4ou. It is armed with a single row of from 16 to 20 hooks which measure about 35 (fig. 7)- There is no neck. YAMB ae lic. 7. Choanotaenia microsoma, n.sp. Head and anterior segments. x 170. INTERNAL ANATOMY Muscular, excretory and nervous systems. As the material was not sufficiently well preserved details of these systems are not obtainable. Genitalia. Testes. There are from 16 to 20 testes situated posterior to the ovary. When fully mature they have a diameter of about 36 (fig. 8). Vas deferens. The genital pore is situated at the extreme anterior lateral angle of the segment and is very large and prominent. The cirrus pouch is short and narrow, extending to the water vessel to which it is dorsal. It lies anterior to the vagina. The cirrus is remarkable in 369 having its extreme tip armed with short spines set at right angles to its length. Immediately median to the tip, the cirrus is armed with a number of hooks of a different shape which measure 30 in length, and which lie Fic. 8. Choanotaenia microsoma, n.sp. Horizontal section showing male and female genitalia. 0.—ovary; 7.s.—receptaculum seminis; s.v.—seminal vesicle; 1.—testes ; v.g.—vitelline gland; +i.v.—water vessel. x 230. S. Fic. 9. Choanotaenia microsoma, n.sp. Showing spines on cirrus. ¢.—cirrus; .p.—genital pore; s.—spine. X 750. ! 5 parallel to the cirrus (fig 9). The vas deferens dilates close to the median extremity of the cirrus pouch into a small seminal vesicle, and then continues in the median direction as a very fine tube (fig. 8). 370 Ovary. This organ lies quite anterior and is divided into two sets of acini, one on each side, widely separated from each other (fig. 8). Receptaculum and vagina. The vagina is a wide muscular tube running posterior to the cirrus pouch and dorsal to the excretory vessel. Near the centre of the segment it dilates into a globular receptaculum, having a diameter of about 36. (fig. 8). Vitelline gland. This is a compact, deeply-staining organ lying posterior to a line joining the two wings of the ovary. It has a breadth of about 110 (fig. 8). Shell gland. This lies immediately anterior to the vitelline gland. It is somewhat globular and has a diameter of about 30.. Uterus. The uterus appears suddenly as a transverse sac situated in front of the ovary. In the next segment the ovary and testes have entirely and as suddenly disappeared, the whole segment being occupied by the uterus which extends beyond the water vessels. The eggs lie in capsules, one in each capsule. DIAGNOSIS The characters which distinguish this worm from other species of the genus Choanotaemia are: (1) its small size; (2) the small number of segments ; (3) the number, size and shape of the hooks ; (4) the peculiarly armed cirrus. On account of its small size I have named it Choanotaenia microsoma. Cyclorchida omalancristrota (Wedl, 1856), Fiihrmann, 1907 Several specimens from intestine of Platalea sp. (spoon bill). Zoological Gardens, Calcutta. Collected by the author, 21.11.13. Sub-family P4RUTERININAE, Ransom, 1909 Rhabdometra tomica, Cholodovsky, 1906 Two specimens from intestine of Francolinus pictus (painted partridge). Zoological Gardens, Calcutta. Collected by the author, 26.3.14. The number of testes and the arrangement of the transverse and longitudinal muscle fibres left no doubt as to the identification of this species. 371 Sub-family HrMENOLEPIDINAE, Ransom, 1909 Hymenolepis medici (Stoss., 1890), Fiihrmann, 1906 Several specimens from intestine of Pelicanus philippensis. Zoological Gardens, Calcutta, 18.9.19. Hymenolepis fusus (Krabbe, 1869), Fithrmann, 1906. 1. A large number of specimens from Larus brunneicephalus. Zoological Gardens, Calcutta. Collected by the author, 22.1.17. The hooks varied in size from 124 to 18. It is important to note that of five worms examined, all of them shewed three or four segments with only two testes. 2. A large number of specimens from Hydropogne caspia (tern). Zoological Gardens, Calcutta. Collected by the author, 17.2.15. In six of these specimens it was found that the number of testes was not constant, many segments possessing only two. Hymenolepis lanceolata (Bloch, 1782), (Weinland, 1858), Braun, 1903 I. Six small specimens 1 cm. long, without heads, from the Black Australian Swan, Chenopis atrata, Berhampur, Bengal, numbered Z.E.V. ae in the collection of the Indian Museum. Collected by Lt.-Col. Clayton Lane, IMS, 11.4.12. 2. About sixteen small specimens 2 to 3 cms. in length from same host. Zoological Gardens, Calcutta. Collected by the author, 24.4.18. 3. About twelve large specimens, 4 to 6 cms. in length and 1 em. in breadth from same host. Zoological Gardens, Calcutta, 24.4.19. 4. About twenty large specimens, about 6 cms. in length and three small specimens, 2 cms. in length from Cygnus atratus. Zoological Gardens, Calcutta, 22.12.19. 5. Four large specimens, 4 to 6 cms. in length from the Black Swan. Zoological Gardens, Calcutta, 25.5.19. The variability of this species is discussed by Maplestone and Southwell in Ann. Trop. Med. & Parasit., June, 1922. Hymenolepis naja (Duj, 1845), Fiihrmann, 1906 1. Three fragments from intestine of Copschychus saularis (Magpie robin). Zoological Gardens, Calcutta. Collected by the author, 5.8.15. All the fragments were stained and mounted. 2. Two specimens, one with a head, from Sita chinensis (green 372 magpie). Zoological Gardens, Calcutta. Collected by the author, 27.4.15. Both specimens were stained and mounted. Hymenolepis zosteropis, Fiihrmann, 1918 1. A large number of specimens from Criniger flaveolus (white cheeked Bulbul). Zoological Gardens, Calcutta. Collected by the author, 26.12.19. Our specimens measured from 2 mm. to 4 mm. in length ; the hooks were very typical of the species. 2. About ten specimens from intestine of Melophus melanicterus. Zoological Gardens, Calcutta. Collected by the author, 25.6.15. 3. Four specimens from intestine of Closa (?) chinensis (green magpie). Zoological Gardens, Calcutta. Collected by the author, 28.4.15. 4. Five specimens from intestine of Ploceus atrigula (the eastern baya). Zoological Gardens, Calcutta. Collected by the author, 12.10.15. 5. Three specimens from intestine of Melophus melanicterus (the crested bunting). Zoological Gardens, Calcutta. Collected by the author, 25.6.15. 6. Six specimens from intestine of Dendrocitta sp. (tree-pie). Zoological Gardens, Calcutta. Collected by the author, 15.5.13., and numbered Z.E.V. °223 in the collection of the Indian Museum. 7 Hymenolepis farciminalis (Batsch, 1786) (R. Blanchard, 1891), Fiihrmann, 1906 Several specimens from intestine of Pica rustica (magpie). Zoological Gardens, Calcutta. Collected by the author, 10.7.18. A striking feature of our specimens of this species was the fact that a single strobila contained segments with no testes, and segments with one, two, three or four testes, although most segments contained three. Another feature was that the testes in some segments were in line, in other segments there were two testes aporal and one poral, and vice versa. In fact their disposition was quite irregular. Hymenolepis stylosa (Rud., 1810), Volz., 1899 1. Several specimens from intestine of Brackypternus aurantius (golden backed wood-pecker). Zoological Gardens, Calcutta. Collected by the author, 31.12.13. 2. Four specimens (only one with a head) from intestine of Trochalopterum meridionale (laughing thrush). Zoological Gardens, Caleutta. Collected by the author, 9.8.15. 373 3. Two young strobilae (2 cms. long) from intestine of Pica rustica. Zoological Gardens, Calcutta. Collected by the author, 10.7.18. These were mounted. Hymenolepis capillaroides, Fiihrmann, 1906 1. Three specimens (one with a head) from intestine of a snipe. Berhampur, Bengal. Collected by Lt.-Col. Clayton Lane, I.M.S., 21.7.12. 2. Twospecimens, one with a head, same host and locality. Collected by Lt.-Col. Clayton Lane, I.M.S., 12.3.12. Hymenolepis (? asymetrica), Fiihrmann, 1918 Three badly preserved specimens, apparently of this species (only one with a head) from intestine of Uvocissa occipitalis (red-billed blue magpie). Zoological Gardens, Calcutta. Collected by the author, 22.10.19 Fiihrmann obtained the species from Chalcococcyx plagosus, New Guinea. His specimens measured 10 cms. in length and I mm. in breadth. The head was armed with 10 hooks 19 long, and of a peculiar shape. Our specimen was armed with exactly similar hooks of the same size. The Indian specimens measured I cm. only in length and were quite immature. In the posterior segments the testes were developing and the rudiments of the ovary could be seen. Hymenolepis (? microcephala) (Rud, 1819), Fiihrmann, 1906 Numerous specimens from intestine of Ciconia alba (white stork). Zoological Gardens, Calcutta. Collected by the author, 6.6.19. Hymenolepis (? simplex) 1. Two fragments and one head from intestine of Tadorna cornuta (sheldrake). Zoological Gardens, Calcutta. Collected by the author, 26.3.15. 2. Numerous specimens without heads from same host and locality, 18.3.14. Hymenolepis spp. 1. A few fragments of a small worm apparently about 12 mm. in length, from the intestine of Emberiza Iuteola. Zoological Gardens, Calcutta, 11.11.15. The fragments were in a bad state of preservation. No head was present ; there appeared to be three testes in the segments examined. 374 2. Other fragments also without heads from intestine of Garrulax belangert. Zoological Gardens, Calcutta, 1.5.10. 3. Still others from intestine of Oviolus melanocephalus. Zoological Gardens, Calcutta, 12.10.15. 4. One specimen without head from intestine of Liothrix lutia (red- billed Liothrix). Zoological Gardens, Calcutta. Collected by the author, 29.5.16. The specimen measured 30 mm. in length and 2-5 mm in breadth. Two testes were situated on one side and one on the other. 5. Fragments from intestine of Dendrocitta rufa. Zoological Gardens, Calcutta, 13.6.15. 6. Several specimens without heads from intestine of Tadorna cornuta (common sheldrake). Zoological Gardens, Calcutta. Collected by the author, 18.3.14. 7. Two specimens without heads from same host and locality. Collected by the author, 23.6.15. In both specimens the testes were irregular, the conditions being similar to those described for H. farciminalis. HYMENOLEPIS ANNANDALEI, nsp. Two specimens from the intestine of Limosa belgicae (black-tailed godwit). Barkuda, Chilka Lake, Orissa, India. Collected by Dr. N. Annandale, 28.4.28. EXTERNAL ANATOMY The specimens had the following dimensions :— Length Greatest breadth Te 60 mm. one att ste I'5 mm, Be 103 mm. ant 300 508 2mm, The anterior part of the worm is attenuated and whip-like; all the segments are broader than long, the posterior and lateral margins being salient. The genital pores all unilateral, and situated slightly anterior to the middle of the lateral margin. Head. The head measures about 180m in length and is 150 broad ; the suckers have a diameter of about 80. The rostellum is a conspicuous organ armed with a single row of 10 hooks which measure about 32 in length (fig. 10). Both in size and shape they closely resemble those of H. brasiliense, Fiihr. The neck measures about 2 mm. in length. Fic. 10. Hymenolepis annandalei, n.sp. Showing head. 220. INTERNAL ANATOMY Muscular system. This is poorly developed. The longitudinal muscles consist of an inner and an outer series of bundles ; the internal bundles are larger and fewer than the outer bundles, the latter being situated immediately beneath the cuticle. A few circular fibres occur between the outer and inner longitudinal bundles and also internal to the inner longitudinal fibres. No oblique fibres were seen (fig. 12). Nervous system. Details of this system were not investigated. A small ill-defined nerve was observed in transverse sections, running external to the water vessel on each side. Water vascular system. This consists of a single ventral vessel on each side, lying ventral to the cirrus pouch and vagina (fig. 12). Genitalia. Testes. There were three testes; one is situated on the pore side and the other two are aporal, one being anterior to the other (figs. Ir and 12). When fully mature they have a diameter of about 150 and occupy almost the whole of the segment dorso-ventrally. Vas deferens. The cirrus pouch lies dorsal to the vagina ; it is some- what club-shaped, the broader extremity being median. It measures about 180 in length and its greatest breadth is about 4ou. Its median half is occupied by an internal seminal vesicle. In the median direction it continues as a very short, wide, coiled tube and then dilates into a large external seminal vesicle which measures about 160m in length and 30m in breadth (fig. 12) ; the median extremity of the external seminal vesicle is close to the poral testis. 376 Ovary. The ovary is situated ventrally in the middle line, and posterior ; it measures about 300m broad and rooy in the antero-posterior direction, whilst dorso-ventrally it practically fills the segment (fig. 11). Receptaculum and vagina. The vagina is a very muscular organ measuring about 450m in length and is club-shaped. At the pore its breadth is about tow; it gradually widens and attains a maximum diameter of 50 at a point opposite the middle of the external seminal vesicle. It then narrows gradually. The whole vagina functions as a receptaculum. Fic. 11. Hymenolepis annandalei, n.sp. Horizontal section showing genitalia. c.p.—cirrus pouch; #.—nerve; 0.—ovary; /.—testes; v.—vagina ; v.d.—yvas deferens ; v.g.—vitelline gland ; w.v.—water vessel. 60. a Im pt. s ¢ ae AM.B-del. Fic. 12. Hymenolepis annandalei, n.sp. Transverse section showing cirrus pouch, vagina and the great development of parenchymatous tissue. ¢.p.—cirrus pouch; ex.ves.sem.— external vesicula seminalis; /.7.—longitudinal muscle; 0.—ovary; .t.—parenchymatous tissue ; ¢.—testes; v.—vagina; w.v.—water vessel. 72. Vitelline gland. This is a conspicuous bi-lobed organ situated posterior to the centre of the ovary. It is about roo broad (fig. 11). Uterus. The uterus consists of a simple transverse sac extending well beyond the water vessel on each side, and almost to the edge of the segment. The eggs were not mature ; the largest measured 17» in diameter and the onchosphere measured Irv (fig. 13). 377 Fic. 13. Hymenolepis annandalet, n.sp. Whole segments showing fully developed uterus. ut.—uterus ; w.v.—water vessel. 35. DIAGNOSIS The worm bears a very close resemblance to H. brasiliense, Fir. The only difference between them is that in Fiihrmann’s species the testes are in a line, whilst in H. annandalei, n.sp., this is not the case. I have pleasure in naming this species in honour of Dr. Nelson Annandale, Director of the Zoological Survey of India. Sub-genus Echinocotyle, Blanchard, 1891 Echinocotyle uralensis, Clerc, 1902 I. One specimen from intestine of snipe. Potsengbam, near Loktak Lake, Manipur, Assam (2600 feet), Station 1. Manipur Survey, 14.2.20. This specimen agreed with Clerc’s description, except that the hooks when isolated measured up to 74. In the type species they measure from 54 to 66y. In some segments testes were entirely absent ; two other segments contained only one in each, and in four or five other segments the posterior aporal testis was absent. 2. One specimen from gut of a snipe. No further data given. 3. One specimen, without head, from Gallinago sp. (snipe). Berhampur, Bengal. Collected by Lt.-Col. Clayton Lane, 12.3.12. 4. Two specimens (one with a head) from intestine of a snipe. Berhampur, Bengal. Collected by Lt.-Col. Clayton Lane, (No. 219b), 17.12.12) 378 Family TAENIIDAE, Ludwig, 1886 Diploposthe laevis (Bloch, 1782), Jacobi, 1896 I. One complete specimen from intestine of Netta rufina (red-crested pochard). Zoological Gardens, Calcutta. Collected by the author, 29.1.14. The specimen was stained and mounted. 2. Fragments from intestine of Nyroca ferina, Chilka Lake, Orissa, India, 24.11.14. Numbered Z.E.V. eli in the collection of the Indian Museum. Diploposthe sp. (2 laevis) A fragment from intestine of Strepsilas interpres (turnstone plover). Chilka Lake, Orissa, 24.11.14. (Chilka Survey). Family ACOLEIDAE, Ransom, 1909 Dioicocestus novae guineae, Fihrmann, 1914 1. Three specimens from intestine of Podiceps albipennis (the little grebe). Zoological Gardens, Calcutta. Collected by the author, 1.5.17. They had the following measurements :— Taste III. “Number Length Breadth Thickness 1. Male ste = ach ae a8 10"0 ccms. 5,0 mm. about Io mm. 2. Male a= a aa ts Beg 10°O cms. 3°5 mm. about 1°o mm. 3. Female 205 and noe we 1770 cms. 5°9 mm. 1°6 mm. The head of the female worm (No. 3) is armed with at least 12 hooks, 320 long (fig. 14). Possibly a few hooks were missing. In shape these Snr al = CSS ~ AMB aet Fic. 14. Dioicocestus novae guineae, Fuhrmann. Showing hook. So. hooks are similar to those figured by Liihe for D. aspera (Mehlis), but in the latter species they measure only 200, to 218m, and are 14 in number. In the male specimens (Nos. 1 and 2) the hooks were missing, but the Species ronat otylus pera vae guineae 379 impressions made in the parenchyma by these hooks were clearly visible. The only trace of genitalia in these two male strobilae consists of two cirrus pouches in each segment, each of which measures 750, in length and 330 in breadth. No spines were seen on the cirrus although carefully looked for. In the female strobila the ovary had almost entirely degenerated. There were a number of gravid segments; the eggs measured about 50 and the onchosphere 28. Four species of this genus are now known (Table IV). Tasie IV. } | | ‘i } Mate FEMALE | Locality Host Rostellum Suckers | Length Breadth | Length Breadth | mm. mm. mm. mm. -| .-.| Argentine Plegadis 70 40 60 eae | Practically Practically | | guarauna absent absent ---| Jamaica, Podicipes 45-130 2-2°5 100-1g0 | 35-49 | Very small Very small | Brazil | dominicus S\_ >= Sahl (p= a a a) ee [LL cee | .-| Europe | Lophaetbyia 280 Gro-g'o | = 340 | S8:o-11°5 | Well developed) Well developed | cristata and | with ry | L. griseigena } / hooks. : | 200-218 y4 | in length . New Guinea | Podicipes novae 60 aon 5° 45 Moderately Moderately | bollandiae developed developed j | with 18-20 | | hooks As the Indian specimens have well-developed suckers and a large rostellum, they are closely related to D. aspera and D. novae guineae. They differ from the former in size and in possessing larger hooks, and agree with Fiihrmann’s description of the latter genus. No hooks were present in Fiihrmann’s specimens. 2. A-second very young female specimen of what I believe to be this species was obtained from the same host and locality, by Dr. Baini Prashad, 2.2.18. The specimen was strongly contracted and was ripe, but no gravid segments were present. It was sectioned and mounted. Cestode sp. A few fragments from Sterna fluviatilus. Zoological Gardens, Calcutta, Sire 15- 380 Order PSEUDOPHYLLIDAE, Carus, 1863 Family DIPHYLLOBOTHRIIDAE, Lihe, IgIo Genus Ligula, Bloch, 1782 Bothria as well as external segmentation completely absent from the larvae ; both develop simultaneously with the maturation of the sex-organs in the definitive host, where the external segmentation which does not correspond with the internal is confined to the anterior end. Longitudinal and transverse muscles irregularly interwoven in the anterior end, posteriorly separated into an inner transverse and an outer longitudinal layer. Type (and only) species: Ligula intestinalis (L). Ligula intestinalis (Linnaeus, 1758) Three larval forms from the coelome of three specimens of Danio acquipinnatus (McClelland), collected by S. L. Hora, Esq., Indian Museum ; Pung-Ka-Mem-John stream, Cherrapunji, Khasi Hills, Assam, 28.10.21, and numbered W 3 and W 4 in the collection of the Indian Museum. Liihe in 1898 arrived at the conclusion that there is only one species of Ligula, and this conclusion was accepted by Linstow in rgor and by Cooper in 1918. The synonomy of both the larval and adult forms is very extensive, and a complete list is given by Cooper (1918). The larval forms occur in the body cavity of Teleosts and the adults occur in the intestine of wading and diving birds. Our specimens are typical in every respect and call for no comment. The author has previously (1913) recorded the occurrence of this larva in the intestine of the following Indian fishes, viz., Labeo calbasu and Nemachilus rupicola. Another larval form, viz., Schistocephalus solidus, occurs much less commonly in the abdominal cavity (and occasionally in the stomach and intestine) of bony fishes, but the larval form of this species is characterised by the fact that it is definitely segmented, and possesses two bothria, whereas in Ligula intestinalis bothria and all traces of external segmentation are absent. 381 REFERENCES Brancnarp, R. (1891). Notices helminthologiques (z), in Mém. Soc. Zool. France, Vol. 1V, p. 420. 8 fi ig. Brocn, M. E. (1782). Abhandlung von der Erzeugung der Eingeweidewiirmer und den Mitteln wider dieselben. 54 pp., 10 pls. Berlin. Cnotopxovsxy, N. (1905). Cestodes nouveaux ou peu connus, I. Arch. Parasitol., Vol. X PP- 332-345- 3 plates. Crerc, W. (1902). Contribution a l'étude de Ja faune helminthologique de l’Oural, I, II. Zool. Anz., Vol. XXV, pp. 569-575, 658-664. 10 fig. — (1903). Contribution a l’étude de la faune helminthologique de l’'Oural. Rev. Suisse Zool., Vol. II, pp. 241-386, tab. 8-11. —_—— (1906). Notes sur les Cestodes d’oiseaux de !’Oural, let II. Ctrbl. Bakteriol., Vol. XLII, (a) pp. 433-436, 532-537; (b) 713-730. : —— (1907). Notes sur les Cestodes d’oiseaux de |’Oural, III. Quelques observations sur Dioicocestus aspera Fihrmann et sur les organes génitaux de Schistotaenia macrorhyncha Rud. Ctrbl. Bakteriol, Vol. XLIII, pp. 703-708, 2 pl. Conn, L. (1899). Zur Systematik der Vogeltaenien. Cutrbl. Bakteriol., Vol. XXV, pp. 415-422. (1899). Zur Systematik der Vogeltaenien, II. Ctrbl. Bakteriol., Vol. XXVI, pp. 222-227. —— (1899). Zur Systematik der Vogeltaenien, III. Zool. Anz., Vol. XXII, pp. 405-408. —— — (1900). Zur Systematik der Vogeltaenien, IV. Cirtl. Bakteriol., Vol. XXVII, pp. 325- — (1900). Zur Kenntnis einiger Volgetaenien. Zool. Anz., Vol. XXIII, pp. 91-98. ——— (1901). Zur Anatomie und Systematik der Vogelcestoden. Nova Acta. Leop. Carol. Akad., Vol. LXXIX, p. 171, 8 tab. Coorsr, A. R. (1918). North American Pseudophylidean Cestodes from Fishes. JI]. Biol. Mon., Vol. IV, No. 4. Crepuin, F. C. H. (1839). Eingeweidewiirmer, Binnenwiirmer, Thierwiirmer. Allg. Encycl. Wiss, Kiinste (Ersch, u. Gruber), Sect. 1, 32, pp. 277-302. Leipzig. Dramare, V. (1900). Paronia carrinoi n.g. sp. von Taenioden mit doppelten Geschlechtsorganen. Ctrbl. Bakteriol., Vol. XXVIII, pp. 846-850. 4 fig. FUurmann, O. (1896). Beitrag zur Kenntnis der Vogeltaenien, II. Rev. Suisse Zool., Vol. IV, 11-132. 7 only lca Zur Kenntnis der Acoleinae. Cirbl. Bakteriol., Vol. XXVIII, p. 363. 12 fig. —— (1901). Neue Arten und Genera der Vogeltaenien. Zool. Anz., Vol. XXIV, p. 271. Druckfehlerberichtigung, p. 320. — (1902). Sur deux nouveaux genres de cestodes d’oiseaux. Zool. Anz., Vol. XXV, p. 357; 2 fig. —— (1905). Das Genus Diploposthe, Jacobi. Curbl. Bakteriol., Vol. XL, pp. 217-224. — (1906). Die Hymenolepisarten der Végel, I. Cutrbl. Bakteriol., Vol. XLI, pp. 352-358, 442-452. 39 fig. ; — (1906). Die Hymenolepisarten der Vogel, II. Curbl. Bakteriol., Vol. XLII, pp. 620-621, 73°-755° ——— (1907). Bekannte und neue Arten und Genera von Vogeltaenien. Ctrbl. Bakteriol., Vol. XLV, pp. 516-536. 43 fig. — (1907). Die Systematik der Ordnung der Cyclophyllidea. Zool. Anz., Vol. XXXII, pp- 289-297. ; — (1908). Nouveaux Taenias d’oiseaux. Rev. Suisse Zool., Vol. XVI, pp. 27-73- 60 fig. — (1908). Neue Davaineiden. Cuérbl. Bakteriol., Vol. XLVII. — (1909). Die Cestoden der Végel des Wissen Nils. Results of the Swedish Zoological Expedition to Egypt and the White Nile, 1901. —— (1909). Neue Davaineiden. Curbl. f. Bakt , Vol. XLIX, Part 1. (1913). Nordische Vogelcestoden aus dem Museum von Géteborg. Meddelanden fran Goteborg, Musei Zoologicka Afdelning. ——— (1914). Ein neuer getrenntgeschlechtiger cestode. Zool. Anz., Vol. XLIV, No. 13. (1919). Notes Helminthologiques suisses. Rev. Suisse de Zool., Vol. XXVII, No. 11. vy. Janicxr, C. (1906). Die Cestoden Neu-Guinea’s. Rés. Expéd. Sc. Néerlandaise Nouvelle Guinée, Vol. V. Jounston, T. H. (1911). New Species of Avian Cestodes. Proc. Linn. Soc., New South Wales. Vol. XXXVI. 382 Krasze, H. (1869). Bidrag til Kundskab om Fuglenes Baendelorme. Dansk. Vidensk Selsk. Skr., naturvid. math. Afd. (5), Vol. VIII, pp. 249-363, 10 plates. ——— (1882). Nye Bidrag til Kundskab om Fuglenes Baendelorme. Dansk. Vidensk. Selsk. Skr., naturvid. math. Afd. (6), Vol. I, pp. 349-366, 2 plates. Krerrt, G. (1873). On Australian Entozoa. Trans. Entomol. Soc., New South Wales. Vol. II, pp: 206-232. 3 pl. KortAn, A. (1920-21). Wégel-cestoden aus New Guinea. 1. Papagei-cestoden. Annals musei nationalis Hungarict, Vol. XVIII. Linnaeus, C. (1758). Systema naturae. 1oth Ed. Holmiae. Linstow, O. von (1901). Die systematische Stellung von Ligula intestinalis, Goeze. Zool. Anz., Vol. XXIV, pp. 627-634. LOnnnere, E. (1889). Bidrag dill kannedomen om i Sverige forekommende Cestoder. Bih. Svensk. Vet. Akad. Hanflingar, Vol. XIV, Afd. 4, 69 p. 2 pl. Lune, M. (1898). Die Gliederung von Ligula. Cirbl. fiir Bakt. Abt. 1, Part 23. (1910). Die Siisswasserfauna Deutschlands. Heft 18. Cestodes. Marotet, G. (1899). Sur deux Cestodes parasites des oiseaux (note préliminaire). C.R. Soc. Biol., Parts. Vol. 1, pp. 935-937: Maprtestonz, P. A., and SourHweELt, T. (1922). Australian Cestodes, V. Ann. Trop. Med. and Parasitol., Vol. XVI, No. 2. Mecerrt, F. J. (1916). A Contribution to our Knowledge of the Tapeworms of Fowls and Sparrows. Parasitology, Vol. VIII. (1920). A Contribution to our Knowledge of the Tapeworms of Poultry. Parasitology, Vol. XII, No. 3. (1921). On two new Tapeworms from the Ostrich, with a key to the species of Davainea. Parasitology, Vol. XIII. Mutter, O. F. (1776). Zoologicae Danicae prodromus, seu animalium Daniae et Norvegiae indigenarum characteres, nominae, et synonyma imprimis popularium. 309 pp. Havniae. Nestopinsky, N. (1911). Zur Kenntnis der Vogeltaninen Mittelrusslands. Centralb. f. Bakt., Abt. 1, Vol. LVII, Part 5. Parona, C. (1898). Elminti raccolti dal Dott. E. Modigliani, alle Isole Mentawei, Engano e Sumatra. Ann. Mus. Civ. Stor. Nat. Genova, Vol. XIX, pp. 102-124. Rarturrr, A., et Lucet, A. (1892). Sur le Davainea proglottina. Bull. Soc. Zool. France. Vol. XVII, pp. 105-106. Ransom, B. H. (1902).. On Hymenolepis carioca (Mag.) and H. megalops (Nitzsch), with remarks on the classification of the group. Studies Zool. Lab. Lincoln, Nebr. No. 47, pp. 151-172, tab. 23-25. — (1905). The Tapeworms of American Chickens and Turkeys. 21st Ann. Rep. Bureau Anim. Industry (1904), pp- 268-285, 32 fig. Surptey, A. E. (1909). The Tapeworms (Cestoda) of the Red Grouse (Lagopus scotecies). Proc. Zool. Soc. London. Souruwett, T. (1913). Notes from the Bengal Fisheries Lab. Indian Mus., No. 1. Fish Para- sites. Rec. Ind. Mus. Calcutta. Stites, C. W., and Hassarr, A. (1894). A preliminary catalogue of the Parasites contained in the Collection of U.S. Bureau of Animal Industry, U.S. Army Medical Museum, Biol. Dept. of the University of Pennsylvania (Coll. Leidy), and in Coll. Stiles and Coll. Hassall. Veterin. Mag., pp. 245-354- Stites, C. W. (1896). Report upon the Present Knowledge of the Tapeworms of Poultry. Bureau of Animal Industry, Bull. 12, U.S. Dept. of Agric., Washington. pp. 1-79, tab. 1-21. Srosstcu, M. (1890). Elminti della Croazia. Soc. Hist. nat. Croatica Ann. 5, Agram, pp. 129-136, 2 tav. : Sweet, G. (1910). Some new and unrecorded Endoparasites from Australian Chickens. Proc. R. Soc., Victoria. Vol. XXIIL. 383 A NEW MALARIA PARASITE OF MAN BY J. W. W. STEPHENS (Received for publication 3 October, 1922) PLATE XVI During the course of experimental work on the treatment of malaria, carried out at the Liverpool School of Tropical Medicine from 1917 to 1921, it was the practice always to control the clinical results of treatment by microscopical blood examinations. Occasionally—perhaps some half-dozen times—parasites other than “ring ’ forms were found in films, and doubt arose as to whether they were quartan or simple tertian. The present paper concerns the parasites found in one such case. Private J. , 188817. December, 1916... ... * Malaria,’ East Africa. January, 1918 Be ... Left East Africa. 8.4.18 to 1.4.18 ... ... The blood films for these dates, made for the and x purpose of counting the leucocytes, are still 27.7.18 to 4.5.18 in existence and show Simple Tertian parasites. 19.7.18 to 20.7.18 ... ... The films still in existence show Simple Tertian parasites. 21.7.18 to 24.7.18 ... ... The entry made in the Card Index was ‘ Negative.’ 28.7.18 wa bce ... The entry was ‘? Simple Tertian.’ 29.7.18 ad are ... The entry was ‘ ? Simple Tertian, ? Quartan.’ A re-examination of these two latter films, still in existence, show peculiar forms. 30.7.18 to 3.8.18 ... ... Owing to the doubt as to the nature of the parasites found, and from the fact that this was not the first time that such doubt had arisen, a series of films approximately at 4-hourly intervals during the daytime was made on the above dates. They were stained for 1 hour with Leishman’s stain, and show perfectly well at the present time the characters to be described. 384 The Temperature Chart of 28.7.18 to 30.7.18 (fig. 1) shows a tertian periodicity, and the parasite findings for these days are as follows : 2327-0 One) arc eee .... Young forms absent. Incompletely segmented forms with 6-8 chromatin masses. 29.7.18 oe as ... Young forms in successive stages of growth during the day. opts CyAaiNe “Soe ... Incompletely segmented forms. Djpel .... Completely segmented forms and young rings. 1.0500) §) 2) Peo cee ... Completely segmented forms and young rings again present. Tuy August — a - ae laa a Elm - Ej a Go Ejm EjM Eim 5 er ’ ae SEE EEE TEE (hy as SHRRBnBRRHBHAe& || PHREBRBBABR BE se i I\ (SHES EA GG Guna Gane la Clea ete Bianes SAAT TAZ HIM ede dasa EET Ee ae cesites fall ele Neg-|S.7._| Ne The periodicity of the parasite appears therefore to be tertian. Although a cycle of development is passed through from 30.7.18 to 1.8.18 the temperature on the latter date reaches only 98-8° F. THE PARASITE Young forms. Small ‘rings’ indistinguishable from ‘rings’ of other species, or round or oval forms with little or no clear area (‘ vacuole’) around the nucleus. No indication of amoeboid activity as judged by irregularity of form. The red cells in which the parasites occur are not uncommonly oval with irregular margins—jimbriated. At this stage the cells are not enlarged and (generally) show no Schiiffner’s dots. 385 Medium-sized forms. These are the characteristic forms. They resemble rather closely quartan parasites in the appearance they present of ‘ solidity’ or ‘ com- pactness,’ and the amount of chromatin and the distribution of the pigment in a lateral band are appearances that recall quartan, but no band-like or ‘ meridional’ forms, as seen in the case of the quartan parasite, were found. They are globular or oval, and occur so frequently in oval red cells that it can hardly be a matter of chance but one of actual significance. In forms with one chromatin mass this is often lateral and roughly triangular. There is a complete absence of the irregular, fantastic, “straggling ’ parasitic forms occurring in cells of not uncommonly twice the normal diameter so characteristic of simple tertian parasites. Schiiffner’s dots are now well marked. Segmenting forms. The gradual transition from young rings to segmenting forms can be traced with ease and certainty. The maximum number of segments (merozoites) appears to be 12. Forms occur with as few as 6 nuclear masses and with the pigment concentrated into a single mass, but it is impossible to be certain in the absence of complete segmentation of the protoplasm whether division is completed. The cell in which these forms lie is either normal in size or slightly enlarged. A slight margin showing Schiiffner’s dots is often seen, and the cell is clearly decolorized. The characteristics then of this parasite so far as concerns the medium forms are a non-amoeboid, pigmented, compact, round or oval parasite, resembling quartan, in a red cell showing Schiiffner’s dots, which is either normal in size or only slightly enlarged. The pigment, so far as can be judged in stained specimens, appears to be brownish black, and granular rather than spicular. A double infection of a red cell was only seen once, viz., with 2 contiguous quarter-grown oval parasites in an oval cell. No forms that could be interpreted as gametes were seen. Now and then, but it has been a rare occurrence, I have encountered a form which I could not distinguish from simple tertian. This parasite appears to resemble that found by Ahmed Emin in 1914 in the case of six pilgrims at Camaran in the Red Sea, and figured and described by him as Plasmodium vivax, var. minuta. I have been unable 386 to procure Ahmed Emin’s specimens for examination, so cannot come to a decision as to the identity of his parasite with the present one. The characters of this parasite appear to me to be different from any of the usually accepted species and I propose to call it Plasmodium ovale. REFERENCE Aumep Emin (1914). Une variété nouvelle du parasite de Layeran. Bull. Soc. Path. Exot., Vol. VIL., p. 385. WK STAM 40 ‘mOITABAIIXA ORE ape stego wnttgeenT’) 388 ; EXPLANATION OF PLATE XVI Plasmodium ovale, n.sp. X 1800 Figs. 1—5. ‘ Ring’ forms Figs. 6—13. Medium forms. Figs 14—22. Pre-segmenting and segmenting forms. ~ ‘ae —— ae Annals Trop. Med. & Parasitol., Vol. XVI aGZr PLATE XVI 18 19 20 21 22 C. Tinling & Co. Ltd., Imp. A. M, Brookfield, del. 389 NOTES ON THE BIONOMICS OF Pe eEGOMIT IA CALOPUS, MEIGcEN, IN BRAZIL PART I BY C. J. YOUNG From the Research Laboratory of the Liverpool School of Tropical Medicine, Mandos, Brazil (Received for publication 21 October, 1922) The following work, which was carried out in Mandos, does not attempt to deal comprehensively with the bionomics of S/egomyia calopus, but is mainly concerned with various points which attracted attention while breeding these mosquitoes in the laboratory. It was undertaken chiefly owing to the noting of some slight differences in the bionomics of this mosquito, as compared with those observed in certain other countries. The factors influencing the bionomics of S. calopus have been shown to be complex, and, owing to its sensitiveness to environ- mental conditions, it is doubtful if conclusions can be drawn from the comparison of experiments which have not been carried out under similar conditions of time, place, etc. The effects of many of these factors may be studied by comparison with controls, the only difference between the control and the subject of the experiment being the factor under investigation. Where applicable, this method was adopted in the following experiments. METHODS The adult mosquitoes were kept in wire-gauze cages, and fed on sugar solution. When eggs were required they were allowed to feed on human blood. In the experiments, eggs and larvae were kept in glass jars on a bench in the laboratory, and those used in any one experiment in which a comparison was to be made were kept together, so that the temperature and amount of light reaching each one were the same. 399 Except where otherwise stated, larvae which hatched out were removed every twenty-four hours. The eggs used in each experi- ment where comparisons are made were from the same batch of eggs, but not necessarily from the same adult, the eggs being mixed before distribution. They were always less than twenty-four hours old at the beginning of each experiment. TEMPERATURE The temperature range in Mandos throughout the year is small. According to official figures from 1902 to 1914, the average of the annual mean temperature was 28°2°C., the absolute maximum 38°6° C., and the absolute minimum 188°C. During the experi- ments the laboratory temperature varied between 33° C. and 24°5°C. The temperature of the water in the jars was always found to be within 2° C. of the atmospheric temperature, and was frequently the same. The daily range of the laboratory temperature was usually less than 6° C. LAYING OF EGGS Fielding (1919) and Bacot (1916) found that females preferred contaminated water to clear for laying eggs in. A few experiments were carried out to discover if this preference existed in Manaos. Watch glasses containing the waters to be compared were placed in the breeding-cage, losses due to evaporation being replaced from time to time. The results, which are given in Table I, are similar to those of Fielding and Bacot. Tarte I. Laying of Eggs in Different Waters. Duration of Experiment Contents of Glass No. of Eggs laid | Percentage of Total 72 days Tap water 945 ae 560 26 River water... a 1590 74 2 Tap water aoe || 563 10°6 32 days River water... ... 1682 315 Water from cesspool 3092 57°9 39? Bacot states that it is misleading to say that the eggs are deposited on the surface of water, as in the great majority of cases they are to be found on the wet margins of the receptacle or other object, and that no instance of an egg being laid on a dry surface was observed. In the following experiment it was found that in captivity the majority of eggs were laid on a damp surface, when available. A watch glass containing rain water and a piece of blotting paper floating on it, the areas of the blotting paper and the water surface exposed being approximately equal, was placed in the breeding- cage. The results are shown in Table II. Taste IT. The Numbers of Eggs laid on Water and on a Damp Surface. Eggs found Duration of Experiment On Blotting Paper | On Water | On Dry Glass | 44 days 1966 | 568 ! $8 No mosquitces were observed laying eggs on a dry surface, and those found dry were probably stranded by capillary attraction and left above water level by evaporation. This could not occur with the blotting paper, as it was floating. Fielding’s findings in Queensland were similar. So far as hatching was concerned, no difference was found between those laid on the moist blotting paper and those laid on water. Wild Stegomyia have been observed on several occasions laying eggs in barrels standing in the enclosure behind the laboratory. The eggs were always laid on the wet sides of the barrel just above the water surface. In giass jars placed outside, eggs were laid on the water, usually near the sides, and many adhered to the jar as the water evaporated. HATCHING OF EGGS Difficulty was at first experienced in getting eggs which were left floating as laid, to hatch. An attempt was, therefore, made to discover the cause of this, and several factors were found to 392 influence the hatching. Those investigated were, position of the eggs (floating or submerged), presence or absence of disturbance, presence or absence of food, and the nature of the water. Many other factors, such as bacterial action, formalin, temperature, humidity, drying, lysol, petroleum, soft soap emulsion, and soap solution, are stated to influence the hatching of the eggs. Many experiments were carried out, all of which are not given below, but the following illustrate the points mentioned. HATCHING OF FLOATING AND SUBMERGED EGGs Fielding (1919) in Queensland, and Bacot (1916) in West Africa, found no difference in the hatching of floating and submerged eggs. In Mandos the difference was definite. Even under the most favourable conditions, floating eggs only occasionally hatched. A batch of fifty-four eggs laid during the previous twenty-four hours were divided into two equal lots. Each lot was placed in a jar containing tap water (7°4 c.c. per egg), rice was added and the eggs were agitated by stirring daily for one minute. One lot was submerged on the first day, and the other left floating for twenty days. The results are shown in Table III. Taste III. Hatching of Floating and Submerged Eggs. Hatched by 2oth day Eggs floating for 20 days. | 27 Eggs submerged on 1st day 27 4 fe) Hatched by 20th day... oat PT Hatched after submergence wee|)) 290% Hatched by 39th day... --s} 96% This result has been confirmed by similar experiments with and without added food, and disturbance, and in rain water, but occasionally a few eggs hatched. Why some eggs hatched when floating, although the majority did not, is not apparent. Under natural conditions the eggs were observed to be 393 submerged by various agencies, chief of which appeared to be rain. In Manaos rain falls at all seasons. Jars containing eggs were exposed to rain with the following results :— 41 eggs out of 56 were submerged by 15 minutes rain. 25 ” ” 26 7 ” ” 5 ” ” 18 ” ” 27 ” ” ” 5 ” ” Eggs laid or becoming stranded on the sides of the receptacle became attached when dry, and when the water rose again remained attached and were therefore submerged. Fully developed larvae usually, but not always, submerged floating eggs, seizing the eggs with the mouth, pulling them below the surface and releasing them. The following results were obtained : — 4 larvae submerged 89 out of 110 eggs in 2 days. Aven rt ds f 19 ,, remaining 21 eggs in 27 days. (sere if. 34 ,, 34 eggs in 24 hours. aD? af 4 4, 27 eggs in 11 days. The reason for the differences shown is not known. Floating eggs were also found to be submerged by insects falling into the water. EFFECTS OF DISTURBANCE ON HATCHING Mitchell (1907), in the United States, records that Duprée found agitation to be a great factor in the hatching, and that if left undisturbed eggs may remain unhatched for over a year. Bacot (1916), in West Africa, failed to obtain a decisive result on this question of agitation, and also (1918) casts doubt on the value of Mitchell’s records. In Mandos the majority of eggs did not hatch unless disturbed. It has already been stated that floating eggs did not usually hatch, even when disturbed. When submerged before they were ready to hatch and left undisturbed, they also usually remained unhatched when no food was added. This is shown in Table IV- 394 Taste IV. Effects of Agitation on Hatching in the Absence of Added Food. Submerged 1st day Control Nature of water No food added Food added Not Agitated Agitated Agitated No. of eggs ... ase 50 30 17 Tap water Hatched in 1 month PA 10% 100%, No. of eggs ... soe 140 | 30 25 Rain water } : Hatched in 1 month O Bain 96% In this experiment two batches of eggs were used, one in tap water and the other in rain water. The control was merely to demonstrate that the eggs were fertile under favourable conditions. The agitation consisted of stirring for one minute daily. The amount of water per egg was the same for each batch. In the following experiment shown in Table V, some hatching took place in the presence of added food, but less so than among the controls. Rice was added to each jar, and all eggs were submerged on the first day. Tasie V. Effects of Agitation on Hatching in the Presence of Added Food. Water 230 c.c. in each jar | Not Agitated Agitated | No. of eggs oe 23 30 30 Rain water S08 | Hatched in 12 days Sc 40% 100% It may be added that a further 30 per cent. hatched when agitation was provided. Various methods of providing disturbance were tried. The dropping of water into the jar so as to submerge the eggs was usually followed, in the presence of food, by the hatching of the majority when they were four days old. Rain had a similar effect. A jar containing twenty-six eggs, four days old, floating in tap water, was placed in rain for five minutes. Twenty-five eggs 395 were submerged, and within four hours eighteen larvae were removed. A control showed no hatching. Stirring or aerating the water for one minute daily, or the addition of one or more larvae, provided an effective stimulus. In the presence of added food, the disturbance caused by larvae appeared to be only slightly more effective than one minute’s stirring, as shown in Table VI. Taste VI. Larvae v. Stirring in the Hatching of Eggs in Presence of Added Food. Two fully-grown larvae present Stirred one minute daily No. of eggs... ae an for 30 No. of eggs ... a5 ae a 30 1oo% hatchedin ... ... ...| 5 days rooUAhatched ince: <5 eees| 8 Gays Each jar contained 230 c.c. of rain water. The eggs were submerged at the beginning of the experiment. In tap water little difference was observed, the larvae hatching the eggs slightly faster. In the absence of added food, however, larvae were more effective, as shown in Table VII. Tasre VII. Larvae v. Stirring in the Hatching of Eggs in Absence of Added Food. Two fully-grown larvae present Stirred for one minute daily No. of eggs... she ae oe 30 No. of eggs ... aes eee 4 30 | Hatched in 18 days ... -... ...| 76% Hatched in 18 days Bea eee len tOoh Total hatched after addition of rice 96% Total hatched after addition of rice | go WE Each jar contained 230 c.c. of tap water. The eggs were submerged at the beginning of the experiment. A similar result was obtained in rain water. In these experi- ments fully grown larvae which pupated or died were replaced by others. It seems probable from these and other results that the larvae, possibly through their excretions, had an effect on the eggs similar to that of the addition of food. 396 EFFECTS OF Foop ON HATCHING In Table VIII it 1s shown that the addition of rice rendered the conditions more favourable to hatching. Taste VIII. Induence of Addition of Rice on Hatching. Rice added fo) 5° mgms No. of eggs in each jar a8 oor a5 18 30 30 Hatched by 2oth day no Ba: sae aha BBs go % Hatched after addition of rice ths ae 2a 83% — The eggs were floating till submerged on the fourth day, after which they were stirred daily for one minute. Each jar contained 230 c.c. of rain water. In other experiments where rice had not at first been used, but other conditions suitable to hatching were present, its addition was invariably followed by hatching, comparison with controls indicating that the hatching was due to the addition of the rice. EFFECTS OF DIFFERENCES IN WATER ON HATCHING Tap water containing more organic matter than rain water might have been expected, on grounds of possible food supply, to be more suitable for the hatching of eggs than rain water. ‘The latter was, however, found to be preferred by the mosquito for laying eggs on, and to be more suitable for the hatching and development of larvae. Tap water consists, in Mandaos, of sedimented river water, an analysis of which, made by Mr. W. J. Debdin, F.I.C., F.C.S., has been published by Thomas (1910). According to this analysis the water contained a considerable amount of albumenoid ammonia, apparently derived from vegetable matter, no nitrates, but B. coli in O'I c.c., and in other ways, resembles what is usually described as a peaty water. Where conditions were favourable for hatching, all, or nearly all, eggs hatched, whether in tap or rain water. In Table [IX are shown the results of an experiment in which the conditions were favourable, 397 there being 50 mgms. of rice in each jar; the eggs were submerged on the fourth day, and two larvae were added to each jar. Tasre IX. Hatching of Eggs in Different Waters under Favourable Conditions. Nature of water (7 c.c. per larva) Rain water Tap water No. of eggs in each jar Roce ee bes) OR 30 30 Hatched by sth day... a hs 35 oct 100% | 93% Where conditions were less favourable, eggs hatched more readily in rain water, as shown in Table X. Here no food and no larvae were added. The eggs were floating till submerged on the fourth day. TaBie X. Hatching of Eggs in Different Waters under less Favourable Conditions. Nature of water (4°6 c.c. per larva) Rain water Tap water No. of eggs in each jar = Ses ai = 50 50 Hatched by sth day... ne tas cH sé: 52% 22% Similar results were obtained under other conditions, and it will be shown subsequently that larvae developed more quickly in rain water. VIABILITY OF EGGS KEPT IN WATER It is well known that eggs will hatch after being kept dry for many months. Bacot (1916) stated that some eggs when kept continually immersed did not hatch for periods of from two to five months. This was tested, and the results are recorded in Tables XI and XII. The eggs were stored in the water in jars, which were undisturbed as far as possible and to which no food was added. At the end of each month shown, a number of eggs were removed and examined, the split and collapsed ones being rejected and the others submerged in rain water to which rice was added and stirred daily. 398 Taste XI. Viability of Eggs stored in Tap Water. Removed after | No. tested Hatched Pupated | Adults hat | = Eggs stored floating ...} 4 months | 20 45% fo) ° 5 months 40 15% | ° ° 6, 7, 8 months 160 ° | ° ° Eggs stored submerged! 4 months 30 46% CRIS | 66% 5 months 30 20% fo) ° 6 and 7 months 30 ° ° ° Tasre XII. Viability of Eggs Stored in Rain Water. | Removed after | No. tested | Hatched Pupated Adults Eggs Stored Floating 3 months 40 92% 75% 75% | 5 months 30 | 40% * 7 months 25 | ° ) One Eggs stored submerged | 3 months 40 100 % EOSivG 95 % 4+ months 40 | 95 % 95 Yo 95% | 5 months 32 56% S 7 months 15 ° co) oe * These observations were kindly made for me by Dr. R. M. Gordon after my departure from Manaos. From each of the four batches used in the experiments shown in Tables XI and XII controls were taken and placed under conditions favourable to hatching, and were found to be fertile to the extent of 96 to 100 per cent. adults being eventually produced. Rejections on account of splitting or collapse of the eggs amounted to 7 to 11 per cent. of the eggs in each jar. Of the adults produced, sixty-four were males and forty-four females. Comparison of the figures in Tables XI and XII would indicate that the eggs retained their viability longer in rain water than in 399 tap water, but such a comparison is not justifiable as different batches of eggs were used, and the times of the experiments, although overlapping, were not identical. Eggs were, therefore, found to be able to remain alive for five months in water, either floating or submerged. This accords with Bacot’s findings in West Africa. Mitchell (1917) records survival immersed at over a year, but gives no details. THE DEVELOPMENT OF LARVAE AND PUPAE The development of S. calopus larvae is influenced by the nature of the water, its amount per larva, the presence of food and its nature, and other factors which were not investigated. In each of the experiments shown in Tables XIII to XVI the larvae used were hatched from the same batches of eggs during the same respective periods, and were all less than twenty-four hours old at the beginning of the experiments. , NATURE OF THE WATER The only waters compared were tap water and rain water. The result is shown in Table XIII. The larvae hatched in the water in which they were subsequently kept. 0°02 per cent. of rice was added to each jar, and the water was aerated daily for one minute by bubbling air through it. Tasrr XIII. Development of Larvae in Tap Water and Rain Water. Water (11 c.c. per larva) Rain water Tap water No. of larvae ... ots aes tee Wes ee 24 24 Pupation commenced a ean es roth day 22nd day Percentage giving pupae... dae ee oo 79% 8% Percentage giving adults... ids es A 79% ° Average larval life of those pupating ee a 19°9 days 22°5 days In the tap water all the larvae became fully developed, but were undersized. The mortality was probably associated in some way 400 with the water, but larvae were quite capable of developing in tap water when less crowded. Fourteen out of fifteen became adults under similar conditions in tap water where the concentration was 50 c.c. per larva. CONCENTRATION OF LARVAE The effects of overcrowding are shown in the following experi- ments, and indicate that where experiments are carried out to test the values of different foods or waters, the results are not comparable if the concentration of larvae has not been the same in each experiment. Tasie XIV. Resuit of Varying the Amount of Tap Water per Larva. Amount of water per larva §0 c.c. | 250 c.c. No; of larvaevinieach\yare cee) seen nee 15 | 15 No. of pupae produced 386 noo ces S60 15 | 15 No. of adults produced... one off | 14(9G33 599) | 15 (1093; 592) Average duration of larval and pupal stages, Jd ... 14°2 days | 10°8 days Average duration of larval and pupal stages, 99 ... 17°8 days | 13°c days This experiment is complicated by the fact that in the jar with 50 c.c. of water per larva there was only one-fifth of the quantity of rice present in the other jar (0'006 per cent.). As it became used up, therefore, rice was added gradually to the former jar till equal quantities had been placed in both without raising the percentage present at any time much above 0'006. Taste XV. Variation of the Amount of Rain Water per Larva. Amount of water per larva is) Cc: 30 C.c No. of larvae ... 385 Se 305 +0 200 20 10 No. of pupae produced oo ose aoe oe 19 10 No. of adults produced oat 250 occ «| 19 (12 gd3 7 99) 10(53d3 5 92) Average duration of larval and pupal stages, Jo... 7:0 days 70 days Average duration of larva and pupal stages, 99 ... 83 days 7°4 days 401 Here the difference is less, possibly owing to more favourable conditions. It was again considered better to provide equal quantities of food per larva in each jar (0°025 per cent. rice and o'006 per cent. peptone) by gradual addition rather than to commence with a double concentration of food in one jar. The water in each jar was aerated daily for one minute. Assuming that the method of adding food did not introduce a fallacy, these and other experiments indicate that overcrowding may influence the rate of development. NATURE OF LARVAL Foop A large number of organic substances have been found to be suitable as food for the larvae, but some appear to be more so than others. In the following experiment peptone and rice were compared. Two jars, each containing 400 c.c. of tap water, were taken, and rice was added to one and peptone to the other to the amount of o'012 per cent. on the first and fourth days of the experiment. An equal number of eggs hatched in each jar during the same period of twenty-four hours. The water was aerated for one minute daily. Details are given in Table XVI. Taste XVI. Peptone v. Rice as a Larval Food. . Food ... 2 soe He ae = aH Peptone Rice No. of larvae... ... “ne i aoe ace 19 | 19 No. of pupae produced... ae aoe i,| 19 | 19 No. of adults produced... nee ase | 19 (1563 499) | 18 (1286; 6 22) Average duration of larval and pupal stages, Sj ..., 71 days 8°6 days Average duration of larval and pupal stages, 99 ... 8'o days g'8 days Thus under these conditions both male and female larvae develop more rapidly on peptone than rice. DURATION OF LARVAL AND PUPAL STAGES The duration of the larval stages varied enormously under the conditions described above. The shortest time observed was four days in the case of three male larvae, the food used being peptone 402 (0'006 per cent.) and rice (0'025 per cent.) in rain water (30 c.c. to each larvae). The longest period recorded was also in the case of a male larva, which did not pupate till the forty-second day after hatching and became an adult two days later; in this case the food was rice alone, and the concentration 9 c.c. of rain water per larva. Macfie (1915) states that under ‘normal conditions’ the larval stage usually lasts seven to thirteen days, and records an instance where it lasted at least ninety-nine days and produced a healthy adult. Bacot (1916) states that under the most favourable conditions the larval life is passed within four days, but with scarcity of food is prolonged for upwards of seventy days. Table XVII gives the average duration of a _ considerable number of larvae living under various artificial conditions in the laboratory. Taste XVII Duration of Larval Stage. Sex No. of larvae Average number of days 14 days and under 6 77 9 99°9 %e 9 48 14°6 62°75 % | Unrecorded 57 73 92'9% | Total 182 9°9 840% The duration of the pupal stage did not vary to any great extent. Figures are given in Table XVIII. Taste XVIII. Duration of Pupal Stage. Sex No. of Pupae | 1 day 2 days ‘| 3 days 3 96 2 $8 6 g 62 2 53 7 Unrecorded | 28 | ° 17 Ir Total | 186 | 4 158 24 403 Of these figures 85 per cent. took two days, and none took as long as four days. Macfie (1915), in West Africa, records the pupal stage as lasting one to five days, as does Mitchell (1907) in WES.A. Table XIX shows the duration of the combined larval and pupal stages. Taste XIX. Duration of Combined Larval and Pupal Stages. No. of Larvae Average number of days | Sex | : 14 days and under 3 | 105 10°3 85°7 % | 2 ) 73 141 / 698 % / | ° Total 178 11°6 79°24 | As these records are based on observations made once daily only, and always about the same time, fractions of a day were not recorded. Individuals stated to have taken any particular number of days may be more or less than the number stated by just under twenty-four hours. SURVIVAL OF PUPAE OUT OF WATER Fielding (1919) placed five pupae on filter paper which was kept moist. All became adults within four days. Of three pupae placed on wet filter paper, allowed to dry and later removed to water, one. hatched after thirty-two hours on the filter paper and two failed to hatch after forty-seven and a half and seventy-two hours, Alcock (1921) records that pupae left on the floor of a cage in London hatched after some delay. In the following experiments the pupae were placed on blotting paper till dry and then transferred to dry glass tubes. 404 Tasre XX. Survival of Dry Pupae. Durat-on of pupal stage Under 24 hours Over 24 hours , before drying Under 48 hours No. of pupae... cho 2 12 3 Result... ac0 a06 eee All died within 48 hours All hatched within 6 hours The dry bulb temperature varied between 29° and 25° C. and the wet bulb between 25° and 23°C. during this experiment. When, however, two lots of dry pupae, four under and four above twenty-four hours, were kept in a dry tube, placed in a stoppered bottle containing water, three hatched out of each lot. It would, therefore, appear that the development of dry pupae was influenced by their age when dried, atmospheric moisture and probably other factors not investigated. MISCELLANEOUS FACTS; RELATIVE NUMBERS OF MALES AND FEMALES The relative numbers of males and females bred in the laboratory are given in Table XXII. Taste XXI. Relative Numbers of Males and Females. Total Adults | No. of Males Percentage of Total No. of Females — Percentage of Total 255 62°5 % | 194 8735 Yo This preponderance of males appeared to be a constant factor under the various conditions employed in the laboratory. REMOVAL OF Eccs By ANTS As Bacot (1916) emphasises the fact that ants in West Africa did not carry off eggs, it is worth noting that in Manaos dry eggs were readily attacked by ants which were observed actually removing them, and batches of dry eggs left exposed on the bench overnight were frequently partly or wholly removed by the following morning. 495 LAYING OF EGGS WITHOUT FEED oF BLoop Fielding (1919) found that eggs were laid when peptone and sugar were given as food without blood. This was tried in Manaos without success, a thick syrup of sugar and peptone being supplied as food in a cage containing about fifty female S. calopus and a larger number of males during a period of one month. Eggs were laid when they were allowed to obtain blood. Mitchell (1907) states that S. calopus will mate and at times lay without feeding. Ort As A LARVICIDE Macfie (1917), discussing S. fascia/a larvae imprisoned beneath a film of oil, describes the efforts made by the larvae to reach the surface. He writes as follows :—‘ So vigorous does the effort appear to be that it seems not improbable that it would eventually break through a thin film of oil.’ The latter occurrence was observed in Manaos in the case ot larvae breeding naturally in a barrel of water, the surface of which was covered with a thin film of oil. The larvae on ascending for air did not usually succeed in penetrating the surface film at the first attempt, but only after repeated efforts at different parts of the surface. The larvae from this barrel were the largest observed, but of some removed to the laboratory only a few reached the adult stage, the majority dying owing to failure to get clear of their larval or pupal skins. The pupae and adults which developed were also unusually large. SUMMARY Various points in the bionomics of S. calopus in Mandos have been investigated, and in certain minor respects they differed from those described in West Africa and Queensland. Bacot (1918), in discussing the duration of viability of the eggs of S. fasciata, writes, “Tt seems to me possible that the African and American races of S. fasciata—to suggest no smaller division—may differ considerably in constitution.’ It is, however, possible that the differences noted in various countries are due to differences in the conditions under which the experiments were carried out, and, therefore, conclusions cannot be drawn from their comparison. -The following facts were found to apply to S. calopus under the particular conditions described above ;— 406 1. The adults laid more eggs in cesspit water than in river or tap water. 2. More eggs were laid on a damp surface than on the water surface. 3. Eggs did not usually hatch when floating. 4. Floating eggs were submerged by rain and S. calopus larvae. 5. Adults laid more eggs, eggs hatched earlier and in larger numbers, and larvae developed more rapidly and showed less mortality in rain water than in tap water. 6. Some eggs stored in water, floating and submerged, hatched after five months. None hatched after seven months. 7. Larvae developed more rapidly and showed less mortality with peptone as food than with rice. 8. The larval stage varied from four to forty-two days and the average duration in one hundred and eighty-two larvae was approximately ten days, the average for the females being longer than for the males. 10. The duration of the pupal stage varied from one to three days, 85 per cent. of one hundred and eighty-six pupae taking two days. 11. Pupae dried at least twenty-four hours after pupation developed into adults, although kept dry. 12. Throughout the experiments more males than females were produced. 13. Dry eggs were removed by ants. 14. No eggs were laid by adults fed on peptone and sugar only. 15. It was observed that larvae were capable of obtaining air through a thin film of surface oil. REFERENCES Atcock, A. (1921). Trop. Dis. Bull., Vol. XVII, p. 118. Bacot, A. (1916). Yellozv Fever Commission Report, Vol. III. (1918). Parasitology, Vol. X, p. 280. Freipine, J. W. (1919). Ann. Trop. Med. S Parasit., Vol. XIII, p. 259. Macriz, J. W. S. (1915). Bull. Ent. Res., Vol. VI, p. 224. (1917) Bull. Ent. Res., Vol. VII, p. 295- Mircuett E. G. (1907). Mosquito Life, Putnam’s. Tuomas, H. W. (1910). Ann. Trop. Med. & Parasit., Vol. IV, p. 7- 407 THE OCCURRENCE OF THE LARVAE OF ONCHOCERCA VOLVULUS (Levcxart, 1893), IN THE SKIN OF NATIVES OF THE GOLD COAST BY J. Fee GORSON (Received for publication 31 October, 1922) PLATES XVII AND XVIII The presence of larvae of O. volvulus in the skin has been described by Montpellier and Lacroix (1920, 1921) and _ by Ouzilleau, Laigret and Lefrou (1921). The former authors stated that the larvae caused itching, resulting in the development of an eruption with papules, vesicles and pustules, which they called filarial itch. The latter authors, on the other hand, considered that the larvae produced an inflammatory reaction in the skin, not especially associated with itching, but giving rise to pseudo- ichthyosis, elephantizsis of the skin of the genital organs and of other parts, leucodermia and atrophy. Brumpt (1920) was not satisfied that the larvae, found in the skin by Montpellier and Lacroix, were those of O. volvulus. The observations of Ringenbach and Guyomare’h (1914), Dubois (1916) and Clapier (1917) are not all in agreement regarding the association or otherwise of elephantiasis with infection with O. volvilus. Larvae of O. volvulus were found in the lymph glands by Ouzilleau (1913) and by Fiilleborn and Simon (1913). They have since been observed by several others. In the blood they have very rarely been found. Numerous examinations have been made by Ouzilleau, Rodenwaldt, Rodhain and Van den Branden, Clapier and Montpellier, Lacroix and Boutin, with almost completely negative results. Rodhain and Van den Branden state that Brumpt, in 1904, found them very rarely, that Rodenwaldt found them once among many negative 408 examinations, and that Ouzilleau found them once only in two thousand examinations. Simon (loc. cif.) found that if he squeezed the finger powerfully in making blood smears, larvae of O. volvulus were present, and suggested that they were in the lymph that exuded as a result of the squeezing of the tissues. The following observations were made on prisoners at Seccondee, Gold Coast. An inspection of two hundred and ninety men was made in order to note the presence of subcutaneous tumours and also of abnormal conditions of the skin, especially a dry, glistening, wrinkled appearance, with exaggeration of the normal pattern of lines and intervening areas, the condition termed lichenification. Elephantiasis and pronounced thickening of the skin, ‘ craw-craw ’ and signs of scratching were also noted. The results are shown in Table I; in the table, L. = lichenification, E. = elephantiasis, T. = thickening of the skin approaching elephantiasis in degree but not specially localised, C.-C. = craw-craw. Taste I. ABNORMAL Skin CONDITIONS Number Subcutaneous tumours present examined | Scratches | | L. | E. Ds CC. present | | | | | 290 | 16 | 24 | I 3 5 20 Twenty-four cases were selected for investigation; of these thirteen had subcutaneous tumours, and in fifteen the skin showed lichenification. Three of the five cases of craw-craw, the three cases of greatly thickened skin, and the single case of definite elephantiasis of the external genital organs, were also included. METHOD OF EXAMINATION FOR LARVAE IN THE SKIN A piece of skin about half a square centimetre in area was excised from the left lower dorsal region in each case and put into a small tube containing normal (0’85 per cent.) salt solution. A bit of the skin was teased and examined soon after excision, the rest being left in the salt solution for a few hours to allow some of the larvae to escape from the skin. The piece of skin was then removed 409 from the salt solution to 70 per cent. alcohol for subsequent section. The whole of the deposit that formed at the bottom of the tube was put on a slide and examined for the presence of larvae, then fixed by the addition of two or three drops of Ruge’s solution (formalin 2 per cent. containing 1 per cent. acetic acid), dried and stained with warm haemalum solution. Drawings and measurements of the larvae were made with the aid of a camera lucida. The chief clinical features and the results of the examination are summarised in Table II, which shows the occurrence in association of subcutaneous tumours, various clinical skin conditions and the larvae of O. volvulus in the skin. The same letters as in Table I are used to denote the skin condition. Taste II. Skin ConDITION Larvae of Number of Tumours O. volvulus cases present Normal) EE. | Ls&E | Es&GC.| Tf. |.C-c. in skin 7 7 4 | 2 I ih 3 3 3 3 5 I I 2 I 5 1 hes I i 2t 2 6 reo 8 6 as | 24 13 5 12 | I | 2 3 I 15 1 * Aspiration of the tumour failed to show larvae of O. volvulus. + Tumours excised and found not to contain O. volvulus. EXAMINATION OF THE BLOOD In each case a thick blood smear was taken from the finger both by day and by mght. In fourteen of the cases a more thorough examination was also made; six or more fresh blood preparations from the finger and from the back near the place from which the excised piece of skin had been taken were examined. The skin was strongly squeezed also in order to see whether larvae of O. volvulus would be readily squeezed out in this way in cases where they were 410 known to be present in the skin. In three cases about 3 cubic centimetres of blood were withdrawn from a vein and centrifuged, and the deposit examined. Larvae of O. volvulus were found in one case only, Case 6, in blood from the skin of the back. The skin in this case was heavily infected, the excised piece yielding about eight hundred larvae to the saline solution 1n which it was put. This case was one of the three whose blood was centrifuged. In the three cases with greatly thickened skin, and in the case ol elephantiasis, thick smears of blood were taken at night from the finger. Embryos of ‘daria bancrojti, Cobbold (1877), were found in the case of elephantiasis. In the course of the examinations cf the blood and of the excised pieces of skin, larvae closely resembling and probably identical with that of Acanthocheilonema perstans (Manson), 1891, were seen in twelve cases, and embryos of I. bancrofti in three cases. VITALITY OF LARVAE OF O. VOLVULUS (a) In normal (0°85 per cent.) salt solution. he larvae deposited in the tubes of salt solution, when examined a few hours after excision of the pieces of skin, were living and showed active move- ment. In cover-glass preparations of teased skin, kept in a moist chamber at room temperature (about 25° C.), the larvae were seen to be alive eight hours after removal from the body; on the following day all were motionless. In a case not included in this series, actively moving larvae of O. volvulus were found in the skin twenty-two hours after the death of the patient from pulmonary tuberculosis. Larvae obtained by aspiration of a tumour in Case 24 and mounted in salt solution under a cover-glass, ringed with vaseline, showed movements for forty-eight hours. (6) In blood. A drop of blood from the skin of the back of Case 6, containing a few larvae of O. volvulus, was covered and ringed with vaseline and kept in a moist chamber at room tempera- ture (25°C.). The larvae showed fairly active movement for over five days. Staining subsequently with haemalum confirmed the identity of the larvae. It is interesting to note that Robles (1919) found that the larvae of Oxchocerca caecutiens, Brumpt, 1919, rapidly died in blood. 411 IDENTITY OF THE LARVAE IN THE SKIN WITH THE LARVAE OF 0. VOLVULUS (@) Morphology. In the few measurements made of living larvae from the skin, the length varied from 290 to 340m, the breadth from 6 to 7“. Stained prepartions showed that they were sheathless, with the cuticle transversely striated for the whole length. The anterior end was free of nuclei for a distance usually of about 10”; the posterior end was sharply pointed, curved generally at a wide angle, and was free of nuclei for usually the terminal 12 to 15. The nuclei were small, mostly oval, longitudinally arranged and closely crowded together, the terminal one being usually distinctly elongated. Of the fixed points of Fiilleborn’s scheme, the ‘ nerve ring’ and last nucleus were the most easily seen and measured. The Gt cel! could not be definitely distinguished in many specimens; in those measured the most frequent position was between 69 and 70 per cent. of the length from the anterior end. Some of these features are shown in Table III, and Tables TV and V give comparative measurements of larvae from tumours. Taste III. Films from the deposit in the tubes containing pieces of skin in salt solution ; measurements of 168 specimens, 12 from each of 14 cases. Length | Relative position of Relative position of “nerve ring’ last nucleus Microns Number | Percentage Number Percentages Number | 235 3 21 3 92°55 = 250 16 22 26 93°5 It 265 46 | 23 82 945 72 280 66 24 52 95°5 78 295 fe Ce 5 96°5 5 ee eS |S ee er ree eee re =. Anterior end free from nuclei Posterior end free from nuclei Microns Number ae Microns | Number “ 6-8 16 ; 7-11 30 g-12 148 12-16 133 13-16 4 17-21 5 412 Tasce IV. Measurements of 50 larvae from a tumour excised from Case 22. Microns Length Number Percentage 6 21 19 22 17 23 5 24 I 25 I 26 27-29 I 30 31 32 Relative position of “nerve ring” Number Relative position of last nucleus Percentage Number Anterior end free from nuclei Posterior end free from nuclei Microns Number | 6-7 | 10 8-9 | 32 | 10-11 8 | Microns Number 30 I! Taste V. Measurements of 40 larvae from fluid obtained by aspiration of tumours in Cases 2 and 7, 20 from each. Relative position of ‘ nerve ring’ Relative position of last nucleus Percentage Number Percentage : 93°5 6 945 27 Che) 3) 3 Number 413 (6) Relationship to tumours. In Table II it is seen that larvae were found in the skin in ten of the thirteen cases with subcutaneous tumours. Of the three cases where tumours were present and larvae of O. volvulus were not found in the skin, tumours were excised in two and found not to contain QO. volvulus; in the third case aspiration of the tumour failed to show the presence of larvae. Hence larvae of O. volvulus were present in the skin in at least go per cent., and possibly in all of the cases with subcutaneous tumours which might be tumours of O. volvulus. By excision in Case 22 and by aspiration in Cases 2, 6, 7 and 24, the tumours were shown to contain QO. volvulus; in these cases larvae were found in the skin. In five cases larvae were present in the skin, but no tumours could be found. These results confirm the observations of Montpellier and Lacroix and of Ouzilleau, Laigret and [efrou, that the larvae found in the skin are larvae of O. volvulus. SECTIONS OF SKIN Larvae of O. volvulus were seen in sections of the skin in the papillary and sub-papillary layers at all levels. In many sections they could be clearly seen to be quite apart from the blood vessels. No very marked changes in the skin were observed; there was an excess of cells in the papillary layer and around the capillaries. Otherwise the sections appeared to show little departure froin normal. THE SKIN CONDITIONS AND THEIR RELATIONSHIP TO THE LARVAE Lichenification was most evident on the back, buttocks and posterior aspect of the thighs; the shoulders and arms were less affected, the chest and abdomen usually stil! less, and the face, throat and limb flexures hardly at all (Plates XVII and XVIII). Sweating of the skin was tested in six of the cases; they were set to do hard work in the sun for a few minutes, and in each case sweating of the affected areas was observed. Itching does not appear to have been severe in most of the cases. One man whose back showed pronounced wrinkling of the skin, 414 maintained that there was no itching and his skin showed no marks of scratching, yet the excised piece of skin yielded about five hundred larvae to the salt solution in which it was placed. The number of larvae counted in the smears of the deposit in the tubes of salt solution varied greatly, the greatest number being seven hundred and eighty-five and the smallest two. In Cases 1 and 12 (Plates XVII and XVIII) the numbers were respectively one hundred and seventy-two and five hundred and forty-three. No relationship between these numbers and the degree of skin affection was established. There were six cases with well marked lichenification in whom the excised piece of skin showed no larvae of O. volvulus. On the other hand, in three cases with larvae in the skin the latter presented a normal appearance. Larvae were and in the four ’ present in the skin :n the three cases of ‘ craw-craw, cases with greatly thickened skin, including the case of definite elephantiasis. In these various conditions there appears to be nothing to indicate a connection between the presence of the larvae of O. volvulus in the skin and the appearances observed. CONCLUSIONS The following conclusions are drawn :— 1. The iarvae in the skin were those of O. volvzlus. 2. They are present in the skin in all, or nearly all, cases with tumours of O. velvulus. 3. No clear causal relationship between the larvae and the elephantiasis and lichenification was ,’ conditions of ‘craw-craw, shown in these cases. 415 40% 30% 20% Percentage of Specimens measured. 10% 21% 22% 23% 24% 25% 26% 21% 28% 8% 24% 25% 26% 40% Percentage of Specimens measured. 10% 21% 22% 28% 24% 25% 26% 27% 28% 29% 80% 31% 39% 93% Fic. 1. Comparison of relative position of ‘nerve ring’ in larvae from (A) skin, (B) excised tumour, and (C) fluid aspirated from tumours; from Tables III, IV and V respectively. 416 REFERENCES Brunet (19201). Bull. Soc. Path. Exot., Vol. XIII, p. 314. (19202). Au sujet des rapports entre l’Onchocerca volvulus et le gale filarienne. Bull. Soc. Path. Exot., Vol. XIII, p. 535. Cuapier (1917). Les porteurs de Kystes filariens (Onchocerca volvulus) et de Nodosités Juxta- Articulaires en pays Toma (Région militaire de la Guinée). Bull. Soc. Path. Exot., Vol. X, p. 150. Dvusoss (1916). Le réle pathogéne de Onchocerca volvulus, Leuckart. Bull. Soc. Path. Exot., Vol. IX, p. 305. Montrecier and Lacrorx (1920). Le Craw-Craw ou Gale filarienne: son origine dans les Kystes sous-cutanés a Onchocerca volvulus. Bull. Soc. Path. Exot., Vol. XIII, p. 395- Monrretiier, DEGouILton and Lacrorx (1921). Note complémentaire sur la gale filarienne et son evolution. Bull. Soc. Path. Exot., Vol. XIV, p. 211. Montretuer, Lacrorx and Boutin (1921). Note hématologique concernant les sujets infestés par Onchocerca volvulus. Bull. Soc. Path. Exot., Vol. XIV, p. 653- Ovzitreau (1913). Les filaires humaines de la Région du Mbonou (Afrique équatoriale frangaise). Pathogénie de |’Eléphantiasis de cette Région. Role de la Filaria volvulus. Bull. Soc. Path. Exot., Vol. VI, p. 80. Ovziteav, Latcret and Lerrou (1921). Contribution a l’étude de /’Onchocerca volvulus. Bull. Soc. Path. Exot., Vol. XIV, p. 717. Rincensacn, and Guyomarc’x (1914). La filariose dans les régions de la nouvelle frontiére Congo-Cameroun. Observations sur la transmission de Microfilaria diurna et de Microfilaria perstans. Bull. Soc. Path. Exot., Vol. VU, p. 619. Rostes (191g). Onchocercose humaine au Guatemala produisant la cécité et l’érysipéle du littoral (Erisipela de la Costa). Bull. Soc. Path. Exot., Vol. XU, p. 442. Ropuain and Van vEN Branpen (1916). Recherches diverses sur la Filaria (Onchocerca) volvulus. Bull. Soc. Path. Exot., Vol. 1X, p. 186. Smmon (1913). Untersuchungen iiber das Vorkommen der Larven von Onchocerca volvulus in Lymphdrusen und in der Zirkulation. (i) Arch. f. Schiffs u. Trop. Hyg., Vol. XVII, No. 233; (ii) Bethefte x. Arch. f. Schiffs u. Trop. Hyg., Vol. XVII, Bethefte 9. (Filleborn and Simon). Reviewed in Trop. Dis. Bull., Vol. III, 1914. be eet ie = ; zs Suivi 4a edd Qi? = itr neta sito WD iy? cthie Sei J a a 418 EXPLANATION OF PLATE "XVII Case 1. Portion of back, showing lichenification of the skin. Three smali tumours were present: cne behind the great trochanter and two over the lumbar vertebral spines. Larvae of O. volvulus were present in the skin. E XVII PIA Annals Trop. Med. & Parasitol., Vol. XVI C. Tinling & Co., Lid., Imp. Case 12. 420 EXPLANATION OF PLATE XVIII Showing lichenification of the skin. No tumours were found. Larvae of O. volvulus were present in the skin. This case had elephantiasis of the external genital organs; the blood contained embryos of F. bancroft2 and of A. perstans. Annals Trop. Med. & Parasitol., Vot. XV 1 PLATE XVIII C. Tinling & Co., Lid., Imp. 421 A CASE OF SLEEPING SICKNESS (T. GAMBIENSE) TREATED BY 7 BAMER .26G BY J. W. W. STEPHENS, AND W. YORKE. (Received for publication 20 November, 1922) Hikes: Aet. 25. No history of having been bitten by a tsetse fly, but in August, 1921, was for 24 days in a tsetse belt at Wamba, South of Jemaa, N. Nigeria. 21.11.21. Took to bed with fever at Jemaa. Ill for 8 days, then recovered somewhat, but still unwell and had aching in legs. —.1.22. Glands in neck found to be enlarged, and a blood film was reported to contain trypanosomes. Was sent home. 9.3.22. On examination at the Liverpool School of Tropical Medicine the condition was as follows :— Lymphatic glands behind the sterno-mastoid on both sides as large as marbles; axillary glands about the same size; inguinal glands (?) enlarged. A circinate rash over the back; over a V-shaped area on the chest, corresponding to the opening in the shirt, deep erythema, with some indication of pitting, probably due to sunburn. Pulse 112. Spleen not enlarged. Blood examination. Fresh films negative. Centrifuged blood, trypanosomes found, I to 10 fields. Gland puncture. A few trypanosomes found. Animal inoculations. 1. Of 2 mice inoculated intraperitoneally with 0-4 c.c. of citrated blood, one became infected 29.3.22 and died 22.9.22; the other did not become infected. 422 2. A mouse inoculated intraperitonealy with a suspension of trypano- somes obtained by centrifuging 10 c.c. of the patient’s blood showed trypanosomes 17.3.22. The animal is stili alive and infected 6.11.22. 3. Various other mice were sub-inoculated from these and became infected. Most of the animals are still alive after six months, and occasionally show trypanosomes in their blood. 4. The trypanosome shows the morphological characters of T. gambiense. Treatment. 10.3.22. Atoxyl, 0-45 gramme, subcutaneously. 13.3.22. Atoxyl, 0-45 gramme, subcutaneously. 17.3.22. Novarsenobillon, 0-6 gramme, intravenously. 20.3.22. Glands smaller, one on right side as big as large pea. Weight, 124 lbs. 20.3.22. Novarsenobillon, 0-9 gramme intravenously. 23.3.22. Patient ill, temperature 103°, blood negative. 27.3.22. Patient better, blood negative, auto-agglutination distinct. 1.4.22. Patient feels well, glands greatly decreased, blood negative, very little auto-agglutination. Weight, 1254 lbs. 8.4.22. Glands hardly palpable, pulse 130, blood negative. 12.4.22. Pulse 108, blood negative, auto-agglutination distinct. Weight, 124 lbs. 18.4.22. Pulse 112, blood negative. Weight, 1263 lbs. 24.4.22. Glands doubtfully palpable, blood negative. Weight, 127 Ibs. ; 29.4.22. Pulse 120, blood negative. Weight, 132 lbs. 6.5.22. Pulse 96, blood negative. Weight, 1364 lbs. I4.5.22. Temperature 103°, blood negative. Weight, 137 lbs. 19.5.22. Temperature 100°. 26.5.22. Thick blood film negative. Weight, 137 lbs. 27.5.22. Temperature 99°. 28.5.22. Temperature ro1-2°. Thick blood film (stained). 2 trypano- somes found. 30.5.22. Temperature normal. ‘ Bayer 205,’ 0-5 gramme intra- venously. Patient vomited a minute or two after the injection. 31.5.22. Temperature subnormal. Pulse 96. Urine no albumen. 423 1.6.22. Temperature subnormal. Pulse 78. ‘Bayer 205,’ 1-0 gramme intravenously. 3.6.22. Temperature subnormal. Pulse 92, a macular rash external to each mamma. ‘Bayer 205, I-5 gramme _intra- venously. 8.6.22. Urine slightly turbid, no albumen. 27.06.22. Temperature normal. Urine slightly turbid, no albumen. “ Bayer 205,’ I-o gramme intravenously. 20.11.22. Patient states that he has remained quite well without any rise of temperature since 27.6.22. Weight, 142 lbs. On eXamination: an acne-like rash over the back and sternum. Glands in neck not enlarged, but some just appreciable to palpation. Pulse 84-86, a little irregular. Respirations 17. Urine, no albumen. Centrifuged blood (5 c.c.) negative microscopically. SUMMARY The patient was presumably infected in Northern Nigeria in August, tg2i, and had no treatment prior to his arrival in England in March, 1922, although trypanosomes had been found in his blood in January. When first seen in Liverpool on 9 March, trypanosomes were found both in the blood and gland juice. He was given subcutaneous injections of 0-45 gramme Atoxyl on ro and 13 March, and intravenous injections of Novar- senobillon 0-6 gramme and 0-9 gramme on 17 and 20 March, respectively. These injections were attended by considerable rises of temperature which lasted up to 23 March. As a result of this treatment the general condition of the patient rapidly improved, the rashes disappeared, the enlargement of the lymphatic glands almost completely subsided, the weight steadily increased, and trypanosomes could no longer be found in the blood. The pulse, however, remained frequent. Except for tworises to 100° F. on 27 Marchand 9g April, the temperature remained normal until 14 May, when it rose to 103° F. Frequent examinations of the blood during this period were negative. On 28 May the temperature rose to ror-2° F., and trypanosomes were found in the blood. On 30 May an intravenous injection of ‘ Bayer 205,’ 0-5 gramme was given, a second injection of r gramme on I June and a third of 1-5 gramme on 3 June: the temperature fell to normal after the 424 first injection, and has since remained normal. " The blood was negative on 31 May and also on 3, 8, and 27 June, and the general condition of the patient remained good. No albuminuria developed. On 27 June an intravenous injection of ‘ Bayer 205,’ I gramme was given as a © prophy- lactic ’ measure, and since then he has remained in good health. We are indebted to Messrs. Friedr. Bayer & Co., Elberfeld, for kindly supplying us with a quantity of ‘ Bayer 205.’ NOTE. In a previous paper by Yorke (1921), details are given of the treatment by ‘Bayer 205’ in July, 1921, of a case of Rhodesian sleeping sickness. The patient, who is now back in Rhodesia, has remained in excellent health up to the present time. REFERENCE Yorke, W. (1921). The Treatment of a case of Rhodesian Sleeping Sickness by the preparation known as ‘ Bayer 205.’ Ann. Trop. Med. and Parasitol., Vol. XV, p. 479+ 425 NOTES ON THE BIONOMICS OF STEGOMYIA CALOPUS, Mzicen, IN BRAZIL PART II BY R. M. GORDON From the Research Laboratory of the Liverpool School of Tropical Medicine, Manéaos, Brazil (Received for publication 9 November, 1922) THE ABSENCE OF STEGOMYIA CALOPUS LARVAE FROM NATURAL WATERS From December, 1920, to February, 1922, a fairly extensive search was made in the town of Mandaos, its native suburbs, and the sparsely inhabited forest surrounding it, for the breeding-places of various mosquitoes. During this examination Stegomyia calopus larvae were never found, except in domestic waters in the immediate vicinity of a human habitation. Young (1921), writing from Mandos, has drawn attention to the same point ; Howard, Dyar and Knab (1912) state: ‘The larvae are found practically exclusively in artificial receptacles about human habitations. It may be said that the larvae of calopus are never found in swamps, in pools or in temporary puddles, even when these are in close proximity to houses.’ The three experiments that follow were devised to test whether the absence of larvae from such waters was due to the disinclination of females to oviposit in them, or to the inability of the larvae to develop when placed there. - Some small stagnant pools situated on the outskirts of the town, about fifty yards from six native houses and about the same distance from the tram line, were selected for the experiments ; water from pools of this description will be referred to in the text as ‘ natural water,’ in 426 contra-distinction to the term ‘domestic water’ as applied to water in rain barrels, water troughs, etc. Experiment I. A varying number (4 to 11) of S. calopus females were confined in two breeding cages, and in each cage were placed six large watch glasses containing water from various sources, both natural and domestic ; the position of these glasses was constantly varied to prevent any undue influence of light or shade. Males were introduced and the supply kept constant. Sugar solution was supplied for the males; the females were fed on human blood, a feed being usually offered every other day. The resultant eggs were removed and counted every twenty-four hours. The experiment was continued for five weeks, with the results recorded below. Taste I. | Total number of | Percentage deposited Nature of water supplied eggs deposited in each type of water 1 | Distilled water ... we 255 =e oe #50 146 9°6 2 | Barrel water in which wild Stegomyia were freely | breeding cas S28 ree a = zeal 88 58 3 Water from a small pool on the outskirts of the town; | | this pool harboured Culex and dragon-fly larvae ...| 149 98 4 | Deep pool near (3); contained Culex but no dragon- | fly larvae a 6 aoe eae ane ore|| 309 20°4 | | 5 | Same as (4), but algae added ... or aoe Sac 728 48°1 6 | Small pool, same source as (3) and (4). Culex and | dragon-fly larvae, but no vegetation oe caH g2 6o CONCLUSION. Séegomvyia in captivity will oviposit as readily in natural as domestic waters. This conclusion agrees with that of Bacot (1916), and of Fielding (1919), but both these authors used domestic water through- out their experiments, and to this added various organic substances. Experiment II. To ascertain whether Stegomyia ova and larvae can develop in natural waters, when these have been cleared of inhabitants inimical to the life of the larvae. Eight jars were used; six of these contained 400 c.c. of water and two 800 c.c., all the waters being carefully strained through fine wire gauze, before the introduction of the larvae. 427 Taste II. No. of Percentage) Average No. No. of larvae No. of of Ova time of Nature of the water used Ova which Imagoes which taken to Jar added hatched completed | complete cycle cycle days 1 | Tap water, plus 2 grs. of rice act 24 23 8 33 28 2 | Water from a barrel in which Stegomyia were freely breeding ... 24 24 fe) 41 24 3. | Smal pool natural water which contained dragon-fly and water- beetle larvae me on 2c 30 21 3 10 22 4 | Small pool natural water which contained no insect life ... 0... 30 26 25 83 21 5 | Tap water plus 2 grs. of rice pe 30 22 3 fe) 37 6 | Small pool natural water which contained dragon-fly and Culex larvae “AE ee nee 30 25 23 70 25 7 | 800c.c. water asin Jar2 ... Bs 30 28 II 36 17 ——| — ————— SSS _— 8 | 800 c.c. water as in Jar 3, plus well- washed duck weed ae ae 30 hr 13 43 10 Conclusion. Stegomyia ova hatch and the larvae develop freely in natural waters after these have been freed from insects inimical to their development. Experiment III, To ascertain whether Stegomyia larvae can develop in pools of natural water when (1) unprotected from their insect enemies, (2) protected from their insect enemies. A small pool such as is described under Experiment I was selected. A careful netting of the pool showed the following inhabitants :—dragon-fly larvae, tadpoles, a few water bugs (Zaitha sp.), a small water beetle (previously shown to be harmless to mosquito larvae). No culicidae larvae were found, though the neighbouring pools showed large numbers, mostly Culex fatigans. Into this pool were introduced goo dried eggs and a few fresh eggs of Stegomia calopus (average fertility of dried eggs was found to be about 40 per cent.), also 300 larvae of Stegomyia and 200 larvae of C. fatigans, the larvae being on an average 48 hours old. Two glass cylinders, 428 arranged as shown in fig. 1, were fixed to pointed sticks and these sunk into the mud at the bottom of the pool, about three inches of the cylinder being left projecting above the surface of the water. Into one tube were introduced 50 dried eggs, and 24 fresh eggs of Stegomyia. Into the other were placed 20 C. fatigans larvae (not more than 24 hours old), During a ne ry Ly oceeee Ce . . Corn Float Fic. I Fic. 2 Fics. 1 and 2. Glass cylinders, 8 in. x 2 in., used in Experiment III. (a = Mosquito-proof gauze; 6 = Fine wire gauze.) the course of the next seven days the pool was regularly visited, but, unfortunately, a week’s heavy rain interfered with the observations and the experiment was brought to a close on the eighth day by the rising water in the pool completely submerging the tubes, 429 The following results were obtained. In spite of very careful searching no Stegomyia larvae were found in the pool during the eight days it was kept under observation, the first search being made twenty-four hours after the introduction of the ova and larvae; C. fatigans were present during the whole experiment, although on two occasions no larvae could be found ; this was probably due to the muddy condition of the water. Of the larvae in the guarded cylinders, both lots appeared to be doing well, three Stegomyia imagoes emerging during the eight days; many of the C. fatigans larvae reached the fourth stage, but none pupated. A few months later the experiment was repeated with the following modifications :—600 calopus and 255 fatigans larvae at all stages of develop- ment were added to the pool, and in the cylinders were placed respectively, 26 calopus ova (average fertility of a sample found to be go per cent.) and 26 fatigans ova (average fertility 100 per cent). In lieu of fixing the cylinders to sticks they were attached to cork floats (fig. 2) and allowed to float clear in the pool. The results obtained were precisely similar to those in the previous experiment, except that no imagoes were obtained though both tubes contained apparently healthylarvae. As before, calopus larvae disappeared after the first twenty-four hours, while fatigans persisted. The observations were brought to a close on the seventh day by the drying of the pool. SuMMARY. Stegomyia calopus ova and larvae, introduced into a natural pool infested with insect enemies, disappeared after the first twenty-four hours, whereas Culex fatigans larvae, added under the same conditions, persisted for at least eight days. S. calopus ova placed in the same pool, but under conditions protecting them from insect enemies, developed and produced imagoes. ORDER OF HATCHING OF MALES AND FEMALES Rees (1gor) states: ‘When mosquitoes are bred in captivity the males as a rule hatch out first, and in greater numbers than the females.’ Nuttall and Shipley (rg0r) comment on this statement as follows : — “We have found no similar statement elsewhere, and the observations we have made do not tend to confirm his observation. The proportion of males to females has always appeared to us to be fairly equal, and we have counted the sexes on several occasions.’ Bacot’s (1916) observations in West Africa would appear to confirm Rees. Writing of Stegomyia he says : 430 . . the early males being usually a day quicker in their development than the females.’ The following note deals only with the order of hatching, Young (1922) having already dealt with the proportion of males to females. As it appeared possible that the food supply might influence the sexes differently, an attempt was made to breed the larvae on different food supplies, other factors being kept as nearly as possible equal. To do this, mixed batches of eggs were sunk and the larvae within twelve hours of hatching trans- ferred to jars containing one of the following two food supplies :— (1) Minimum food supply, viz., tap water to which was added 0-015 per cent. polished rice and 0-5 grm. well-washed duck-weed to each 300 c.c.’s. water. (2) Maximum food supply, viz., stagnant river water filtered through fine wire gauze, to which was added o-o18 per cent. Peptone (Fairchild) and 0-5 grm. well-washed duck-weed to every 300 c.c. of water. Each larva was allowed 30 c.c. of the prepared water, this amount being regulated every day; thus to start with, ten larvae were placed in 300 c.c. of the food supply ; if two died within twenty-four hours then the amount of water was reduced to 240 c.c. and so on. The results are shown in the following tables :— Taste III. Maximum food supply. Number of larvae = 270. Day of | | emergence ...| 5 6 | 7 8 9 10 II 12 3 14 15 | Total Malesjris.iue we ° 4 ° wv ° | 17 ae 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 | 431 SumMARY. Ina mixed batch of ova, hatched and allowed to develop under food conditions which were either (1) favourable, or (2) adverse to growth, it was found that a much greater number of males than females reached maturity during the first few days of the emergences. This preponderance of males was greater than could be explained by the higher proportion of males to females (142 to 98, and 105 to ror) as observed in the completed experiments. OVULATION Experiment I. Results of Diets other than Blood. Goeldi (1905), in Brazil, after numerous experiments with fruit, sugar, honey, etc., came to the conclusion that blood was necessary for the pro- duction of eggs by Stegomyia calopus. Fielding (1919), working in Australia with the same species of mosquito, obtained fertile eggs on three occasions on which peptone and sugar was given as a food. Ken (r917), in India, fed Stegomyia scutellayis on sugar, milk and sugar, peptone and sugar, with positive results. : S. calopus females were kept under observation for a period of over twelve months, at least twenty being always present in the cages. During this time the ordinary food supplied was sugar and water, and on two occasions a mixture of sugar and peptone was given for several days ; the results were similar to those already published by Young (1922), no instance of egg laying being recorded. Several authors record mosquitoes feeding on plants; thus Theobald (rgor), states: ‘I have frequently seen Culicidae settled on Compositae sucking the juices of the flowers, both males and females,’ and Giles (1902), states: “When mosquitoes are unable or unwilling to obtain blood they suck the juices of plants.’ Knab (1907), quotes other instances. The following two experiments were made to see whether Stegomyia would feed on flowering plants, and if so, whether ova would result. (x) Thirteen females were confined for thirty-eight days in a cage and supplied with water and a variety of flowering plants, representing as nearly as possible all the species growing within a ten-yards’ radius of a heavily infected breeding place (a disused water barrel) ; in all seventeen species of plants were used, and each plant was allowed to remain in the cage for three to four days. In addition to the plants the fruits ‘ Goiaba’ and banana were supplied. Males were always kept present. Both males and females constantly alighted on the flowers, inserted their proboscis in the corolla, and apparently absorbed some fluid. 432 (2) Seven females were observed under the same conditions for twenty- one days, but the following additional fruits were used : Melon, ‘ Mammiao,’ Mango, Orange, ‘ Periba,’ ‘ Caju,’ the results being the same as in the first experiment. A trial was then made of the following native fruits : (3) Three females fed for thirteen days on Mango, ‘Mammiao,’ Melon, Orange, Banana. (4) Five females fed for thirteen days on Mango, Orange, Banana, ‘ Periba,’ “Mammao.’ No eggs were laid in either of the latter experiments. SumMARY. Female Stegomyia were offered and fed readily on sugar, sugar and peptone, flowering plants, and various fruits. No eggs were laid after feeding on any of the above substances. Experiment IT. Results of feeding on animals and birds with special reference to Bats and Wall Geckos. Durham (1902), MacGregor (1915), Bacot (1916), Theobald (1916), and Fielding (1919), record Stegomyia feeding on Dog, Goat, Rat, Bandicoot, Agouti, and Guinea Pig; the results of the author's experiments of feeding S. calopbus on various animals and birds are recorded in the following table. Taste V. { | Whether Whether No. | Animal or bird used and method of feeding adopted. seen to | Ova laid fertile | feed | if 1 | A small finch confined in mosquito cage day and night on eight. occasions aA aoe ae Bs eaH 3 No ° = 2 | A rock-dove confined in mosquito cage day and night on four occasions ... = = oS So5 208 2S No | ° = 3 | Young parrots; the tube containing the mosquitoes was | applied to the host’s body... BAS oe och ee] ees 55 = 4 | Domestic chickens; method of feeding as above... al Yes aa + 5 | A young otter; method of feeding as above. (Only a little blood absorbed, partly due to restlessness of animal) OL Ip pe MACE ° = 6 | Lesser Ant Bear; method of feeding as above. (Only half- hearted attempts made to pierce the skin) ... ee ees No ° = 7 | Monkey; method of feeding as above a = cae Yes =f et 8 | Cotia; method of feeding as above ... ne aa0 pes} wes =e ae g | Young Iguana (Urocentron azureum) confined in mosquito cage for some days and nights oa ern hoe ---| No ° = 10 | Young Wall Gecko, placed loose in cage and also enclosed in tight fitting gauze bag; several trials day and night a3 No | ° - 11 | Bats (Molossus obscurus) left loose in cage; several experi- ‘ ments tried both day and night__.... 208 BNO olin ease + Se 433 The above list requires no comment, except for the last two animals named. In all houses observed in Amazonas, whether deserted or occupied, two animals were constantly found present, viz., the gecko and various species of bats. Special attention was, therefore, devoted to seeing if Stegomyia would feed on these in the absence of human blood. The experiments with the gecko were frequently repeated, using both young and adult specimens. At first it was allowed loose in a cage of hungry mosquitoes, none of which attempted to bite. The gecko destroyed numbers of the mosquitoes, so in subsequent observations it was enclosed in a tight-fitting gauze bag and placed on the bottom of the cage. Though mosquitoes were often seen to alight on the gauze and probe it tentatively, they were never seen to draw blood, nor were any females gorged in the morning if the gecko and bag were allowed to remain in the cage over night. The only record noted of mosquitoes feeding on bats is that of Durham (1902) at Para, who observed a Stegomyia calopus female feed on ‘a small bat (Phyllostoma).’ The following species of bats were found to be common in or around houses in Mandos : Saccopteryx bilineata, Tenum., Hemuiderma perspicillatum, L., Vampyrops zarhinus, H. All., Molossus rufus, Geoff., Uroderma bilobatum, Pet., Molossus obscurus, Geoff. Of these Molossus obscurus appeared to be the commonest in houses, and was used in the following three feeding tests. (1) Eleven offered a feed and nine fed ; (2) three offered, two fed; (3) six offered, six fed. Not only did a far higher percentage of those given the opportunity feed on bats than on other animals, but they appeared to attack their host with a far greater voracity than they were observed to exhibit towards any other creature except man. They usually settled on and pierced the wing membranes, and as soon as they were flicked away returned to the attack, the complete feed being thus performed in a series of interrupted bites. ConcLusIon. Stegomyia calopus feeds with great readiness and voracity on bats; it appears likely that these serve as important food reservoirs in deserted houses, or sparsely inhabited districts. It will also feed, but with less eagerness, on certain other animals and birds. All attempts to induce Stegomyia to bite the common wall gecko failed. Experiment III. Results of feeding on washed red cells, serum, and citrated blood. Otto and Neumann (1905), in Brazil obtained fertile eggs by feeding Stegomyia on blood and salt solution. Bacot (1916), in West Africa, on 434 two occasions obtained single eggs by feeding the mosquitoes, on one occasion on honey and blood, and on the other on syrup and blood, one of these eggs proving fertile. Marchoux and Simond (1906), at Rio imprisoned eight female Stegomyia and fed them as follows: Two on fresh human Taste VI. No. of No. of No. ot No. female days Food: and how offered Ova laid Stegomyiae | observed 1 | 6 15 Washed sheep’s cells in normal saline. 0°5 c.c. in a watch- elas’) 2 = <3 oss ee ° 2 5 17 Washed sheep’s cells in normal saline. 05 c.c. in a watch- glass ° 3 | 7 19 Sheep’s serum. 0°5 c.c. in a watch-glass 4 7 24 Washed human red cells in normal saline. o°5 c.c. in a watch-glass ° 4a 3 9 Washed human red cells in normal saline in tubes; same mosquitoes as in (4) ° 5 6 26 Human serum. 0°5 c.c. in a watch-glass ° 5a 3 5 Human serum, in tubes; same mosquitoes as in (5) ° 6 6 29 Whole human blood in saline. 0°5 c.c. in a watch-glass ° 6a 3 8 Whole human blood in tubes with normal saline; same mosquitoes as in (6) ... one ta 3: as oe ° 7 3 10 Cotton ball soaked in whole human blood plus normal saline, suspended in cage... +5: 595 Bee = aa ° 8 | 3 16 Cotton ball soaked in washed human red cells in normal saline, suspended in cage... ae ae = ees x ° 9 3 13 Cotton ball soaked in human serum suspended in cage aoe ° 10 2 I4 Cotton ball soaked in washed human red cells plus normal saline, suspended in cage... Bee ace i] FAHRENHEITS SCALE Case of Trypanosomiasis: Chart. 459 OBSERVATIONS ON ONCHOCERCA VOLVULUS BY J. W. S. MACFIE AND J. F. CORSON (Received for publication 28 November, 1922) The following brief and somewhat disconnected notes on Onchocerca volvulus are based on observations made at Accra in the Gold Coast, West Africa. DIAGNOSIS. The tumours of QO. volvulus are by no means always large and easily recognisable, but are frequently very small, deep-seated, and difficult to detect. In some cases, indeed, we have been able to palpate them only after they had been located for us by the patients themselves. The diagnosis of volvulosis by the presence of tumours is, therefore, unreliable, and we have found it more satisfactory to examine the skin for larvae. The method we adopt is to remove from the lower part of the back a small piece of skin similar to, but rather larger than, those used in skin-grafting by Reverdin’s method. The skin is raised with the point of a needle, and a piece of the required size snipped off with a pair of sharp scissors. The pieces of skin may be examined immediately by teasing them on a slide with a little normal saline solution, or they may be left for an hour or two in saline solution in small tubes, in which case the larvae will be found to have worked their way out and to be lying at the bottom, or they taay be used for sectioning. The technique is simple and rapid, and as it is not painful and is not objected to by African patients, is capable of wide application. The little wounds heal rapidly. In such pieces of skin removed from patients with O. volvulus tumours we have invariably found larvae. In the one or two apparent exceptions met with, the tumours on removal proved to be juxta-articular nodules, and not O. volvulus tumours. In many other cases in which no tumours could be found, the pieces of skin removed in this way contained QO. volvulus larvae. As regards ‘lichenification’ and the other skin conditions 460 sometimes considered to be due to volvulosis, we have on the one hand observed them in skin in which no larvae were found, and on the other hand found larvae abundantly present in apparently normal skin. Moreover, we have recently found another filarial larva in the skin which, at Accra at any rate, is even more commonly present than that of C. volvulus. In view of this discovery, further observations are necessary before it can be said if either of the two larvae is responsible for the lesions. INCIDENCE. In order to obtain some idea of the prevalence of O. volvulus infection in the Gold Coast, fifty men, taken at random, were examined at Accra, all of whom were adults, between the ages of 25 and 45 years, who appeared to be in good health. The examinations were made on the 24th of October, 1922, between the hours of 9.45 and 10.15 a.m. From each man a small piece of skin, as described above, was removed from the small of the back and placed in a tube containing about 2 c.c. of normal saline solution. The piece of skin was subsequently teased up together with a drop or two of the saline solution from the bottom of the tube, the preparation fixed by heating, dried, and stained with haemalum. In no case was there obvious blood in the specimen. The result of this examination was that larvae of O. volvulus were found in seventeen of the men (equal to 34 per cent.). PERIODICITY. Ten of the men referred to above, in whom larvae had been found, were re-examined two days later at about 9 p.m. No sensible difference was observed suggestive of a periodicity in the prevalence of the larvae in the skin. This observation is in harmony with that of Montpellier and Lacroix (1920). DISTRIBUTION OF THE LARVAE IN THE Bopy. In most of the cases examined we have sought for the larvae of O. volvulus in the skin of the lumbar region or the small of the back only. In a few instances, however, we examined other parts also; for example, in a Kru man with a small tumour in the left inguinal region, larvae were found abundantly in the skin of the left buttock, the right ankle, the right shoulder cap, and the right wrist. Our observa- tions, indeed, showed clearly that even in subjects in whom no tumours could be detected and whose skin was normal, larvae of O. volvulus might be found in the skin of widely separated regions of the body. 461 ‘TABLE The distribution of O, volvulus larvae in the body. I | II IIT Parts of the body examined Kruman, | Ashanti man, | Ashanti man. c. 25 c. 40 c. gO 2S I Skin of scalp : left occipital region nil — = above the right ear —- — nil Skin, behind the left ear ... — | mf. v. numerous —- = | — nil Skin : right wrist ... left wrist Skin, second finger of right hand Skin, small of back Skin, scrotum Skin : right ankle ... left ankle Mucous membrane of mouth, lower lip... Stomach Small intestine Large intestine Rectum Mesentery ... Parietal pleura, 8th interspace Intercostal muscle, 8th interspace Lung Ty Heart, left ventricle Aorta ose PETItONEUNE 1 acc.) sce Liver at Spleen RAGS WM even feds.) ofl ass Bladder... Brain : cerebral cortex cerebellum Lymphatic glands: near aorta ... mesenteric... inguinal | mf.o. m| .v. ! mf.v. numerous numerous numerous hil nil nil nil mf. v. numerous mf.v. few mf.v. numerous nil mf.v. = larvae of Onchocerca volvulus. nil = no larvae found. — = not examined. 462 In order to ascertain more accurately the distribution of the larvae in the body, three natives were examined particularly in the mortuary, the first a Kru man, aged about 25 years, and the second and third Ashanti men, aged about 40 years. The first two men had died from pulmonary tuberculosis, and the third from cerebral congestion. No tumours were found in any of the three, and no definite skin lesions of the types associated with volvulosis, excepting in the third man, who had slight ‘ lichenification’ of the back. The various parts of the body examined, and the results, are shown in the accompanying table. About 0°25 c.c. of each tissue was examined. It will be noted that larvae were found in the skin of widely separated areas, but that they were not found in any of the mucous membranes or organs. During the autopsies a careful search for adult worms was made in the mesentery and the retro-peritonea! tissue in the neighbourhood of the liver, the duodenum, and the aorta, but none were found. The inner surface of the aorta (in view of the fact that O. armillaia is abundant in this situation in cattle at Accra), and a number of lymphatic glands from the mesentery, the inguinal region, and near the aorta were also examined without success. LARVAE ARE NOT FOUND IN SWEAT. Although the larvae of O. volvulus are abundant in the skin, they do not appear in the sweat. One of the laboratory staff, a Mendi, in whose skin larvae - were numerous, was set to work in the sun until he perspired freely. Sweat was then collected from his face, chest, abdomen and back, and examined for larvae. None were found. EXPERIMENTS WITH TSETSE-FLIES. Leiper (1913) failed to trace any development of the larvae of O. volvulus in Stemoxys calcitrans and S. nigra, and Rodhain and Van den Branden (1916) failed with Stegomya fasciata and Cimex rotundatus. Brumpt has suggested that the larvae may develop in a tsetse-fly, but so far as we are aware, no observations have yet been recorded in support or otherwise of this view. A few experiments were, therefore, carried out at Accra, in which wild tsetse-flies were fed on patients in whose skin O. volvulus larvae were abundant, and subsequently dissected and examined for developmental stages of these parasites. Unfortunately for our purpose Accra is situated in an extensive 463 tsetse-free area, and we were, therefore, able to procure only a few living flies for our experiments. Glossina palpalis, R.D. ‘Three flies were fed once only on a case of volvulosis and dissected, two on the sixth day, and one on the seventh day after the infecting feed. No filarial larvae were found. Ten specimens which had not been fed experimentally were also dissected as a control. No filarial larvae were found in them. G. longipalpis, Wied. Six flies were fed once only on a case of volvulosis and dissected, two on the twelfth day, and one each on the second, fourth, fifth and sixteenth days after the infecting feed. No filarial larvae were found. Fifteen specimens which had noi been fed experimentally were also dissected as a control. No filarial larvae were found in them. These few observations do not support Brumpt’s suggestion, so far, at any rate, as concerns G. falpalis and G. longipalpis. The number of flies employed was, indeed, regrettably small, but if volvulosis is as prevalent as the figures we have given suggest, and if the parasites are able to develop in them, it might have been expected that one or two of these tsetse-flies (including the controls) might have shown them. EXPERIMENTS WITH LICE. From the usual position of the larvae in the skin, namely, close under the rete mucosum, we are inclined to think that the intermediate host, if indeed it is a biting insect, will prove to be one which does not probe the skin deeply. Lice at once suggest themselves, but so far as our observations have at present proceeded we are not able to incriminate them. One of us (J. F.C.) dissected about sixty specimens of Pediculus humanus corporis at Sekondi without finding any filarial larvae, and further dissections (forty-six) at Accra have been equally fruitless. Moreover, twenty lice taken from the bodies of two men not infected with O. volvulus and fed on a man who harboured larvae of O. volvulus in his skin, and larvae of Acanthocheilonema perstans in his blood, were dissected an hour later. Larvae of O. volvulus were not found in any of them, but living and active larvae of Ac. perstans were observed in eight. This experiment suggests that the lice, in feeding, draw up the larvae present in the blood of their host, but not those in his skin. Jt may be added that in several of the lice dead and partly digested larvae of Ac. perstans were found 464 (derived, presumably, from some previous host), an observation which confirms that of Low (1903), who failed to trace development of this worm in P. capitis and P. vestimentorum. From the fact that the larvae are particularly numerous in the skin at the base of the trunk (buttocks, scrotum, &c.), Phthirius pubis might be regarded as a likely host. Contrary to expectation, these creatures have proved difficult to obtain at Accra, and we have not yet been able to procure any for dissection and experiment. REFERENCES Leper, R. T. (1913). Report of the Helminthologist, London School of Tropical Medicine, for the Half-year ending April 3oth, 1913. Report to the Advisory Committee of the Tropical Diseases Research Fund. Low, G. C. (1903). Filaria perstans. Four. Trop. Med. S Hyg., Vol. VI, p. 199. Monrpe tier, J., and Lacrorx, A. (1920). Le Craw-craw ou gale filarienne; son origine dans les kystes sous-cutanés a Onchocerca volvulus. Bull. Soc. Path. Exot., Vol. XIII, PP-139553%5< Ropuarn, J., and Van pEN Branpen, F. (1916). Recherches diverses sur la Filaria (Onchocerca) volvulus. Bull. Soc. Path. Exot., Vol. 1X, pp. 194-196. 465 A NEW SPECIES OF FILARIAL LARVA FOUND IN THE SKIN OF NATIVES IN THE GOLD COAST BY J. W. S. MACFIE AND Jj. F. CORSON (Received for publication 28 November, 1922) When investigating the occurrence of larvae of Oxchocerca volvulus in the skin of natives in the Gold Coast, sheathless larvae of another species of Filariidae were found in several cases. So far as we are able to ascertain these !arvae have not previously been noted, and, therefore, notwithstanding the fact that we have not yet discovered the adults, a brief description of them is given here. The larvae were found in the skin of nine out of twenty-four cases selected for examination for O. volvulus larvae either because they had tumours, or because the skin showed the conditions which have been associated with that infection. From each of these cases a piece of skin, about half a square centimeter, was removed from the small of the back and placed in a tube containing normal saline solution. The larvae, which were found in the deposit which collected at the bottom of the tubes, were fixed by adding Ruge’s solution to some of the deposit on a slide, allowed to dry, and subsequently stained with haemalum. ‘The larvae were also found in the skin of the forearm and back in one out of nine unselected autopsies. In this case the larvae were fixed on a slide by heating, dried, and stained with eosin-azur. The description of the larva which follows is based on the examination of specimens from these ten cases. Seventy-two larvae were measured, ten from each case in which this number could be found, and all that were available in the others. It may be noted here that there was a certain degree of variation in the size of the larvae in different cases, in some they were on the average slightly larger than in others. 466 MORPHOLOGY. The larvae are sheathless, slender, tapering both anteriorly and posteriorly, and when fixed assume a characteristic form, the body being straight, or nearly so, excepting at the posterior extremity, which is curved round like the handle of a walking-stick (see fig. 1, A). The cuticle is striated. The nuclei are sssesss ste “so 08000y =, Fic. 1.—A. The larvae, x c. 150, to show the general form; Band C. The anterior and posterior extremities X c. 1375; Dand£E. The anterior and posterior extremities of the larva of Ac. perstans, X c. 1375, for comparison with B and C. rather large, two or three abreast in the middle of the larva, and completely fill the greater part of the body. Length. The lengths of the seventy-two larvae measured ranged from 180m to 240@, average 215°5m. The table shows that nearly 60 per cent. were between 210m and 22Qh. Breadth. The breadth at the widest part of the body is about 3p. Anterior extremity. The body tapers very slightly towards the anterior extremity, and is bluntly rounded at its end. No ‘fang’ could be distinguished. The clear area at the anterior end is about 4# long. The column of nuclei commences with a single row of ten 467 or twelve nuclei, the first four being usually oval and the others somewhat quadrate. The distribution according to lengths, and to the position of the nerve ring, of seventy-two of the filarial larvae. Nerve Ring : Lengths, in microns distance from anterior extremity, in microns. 180m to 189... ae eae “5 5 4outo44u ... we ae oe _ Igo to 199M... om Ace a 6 454 to4gu ... Fi ok es 2 200fL tO 209M... se eee 7 som tos4u ee. 5c Sct ace 17 2104 to 219M... cca vee aa 22 s5uto sou... or ase a3 28 220p4 to 229)... rer sc toe 21 Go to 64fe ae San <5: 22 230 to 239u wks one oe 10 65 tob6gu ... wee wee oe 2 PROTEOMMOIE EAD 422.) Ml cents Wace I OIMLOUZA WE Weoe «0% eee a0 I Other anatomical fixed points. The nerve ring 1s situated about 269 per cent. of the length from the anterior extremity: it is a well marked break, in the middle of which is a single, prominent nucleus. In the seventy-two individuals measured, its position varied from 48 to 7Im, average 58 from the anterior extremity. The excretory pore is small, and is situated about 34°1 per cent. of the length from the anterior extremity; the excretory cell lies slightly more posteriorly. The Gr cell, which is not always easily recognised, is large, with a round nucleus, and situated about 69'2 per cent. of the length from the anterior extremity. The anal pore is a small break in the column of nuclei situated about 862 per cent. of the length from the anterior extremity. A central viscus was not seen. Posterior extremity. The body tapers for a considerable distance towards the posterior extremity, and the extreme tip of the tail, beyond the last nucleus, is abruptly pointed so that the posterior clear area is at most about 1m long. The tail is curved sharply into a crook, and the column of nuclei at its extremity is a single row of rounded, or at most oval, nuclei. SITES WHERE THE LARVAE WERE FOUND. The larvae were found only in the skin. The part examined in nine of the cases was the 468 small of the back, and in one case the right forearm and the back between the blades of the scapulae. In the latter case skin from the abdomen near the umbilicus and from the middle of the outer side of the calf of the left leg were also examined, but no larvae were found in these situations. In nine cases six or more blood films from the finger and from the back (near to the spot where larvae were found in the skin) were examined; and in two of these four thick films taken at night, and in two others 3 c.c. of blood taken during the day, were also examined. In none of these specimens were the larvae found. Larvae of Acanthocheilonema ferstans, however, were found in two. In one post-mortem examination in which the larvae were found in the skin of the small of the back, the following parts of the body were also examined, but without discovering any larvae: skin of scalp above the right ear, skin of right wrist, skin of left ankle, skin of scrotum, mucous membrane of the mouth, rectum, lung, aorta, liver, spleen, cerebral cortex of brain, cerebellum, and lymphatic glands along the aorta, in the mesentery, and in the right inguinal region. Sections of the skin showed the larvae lying in the tissue spaces of the cutis vera or corium, usually close to the rete mucosum. There was present in all the cases examined a slight degree of cellular infiltration, especially round thé blood vessels, but with this exception no definite departure from the normal condition was observed. PATHOGENICITY. Our observations, which were made in the course of an investigation of velvulosis, do not admit of any state- ment being made as to the effects which may be caused by infection with this parasite. It may be noted, however, that the condition of the skin known as ‘lichenification’ was present in six of the ten cases examined, and a definite thickening in two others. Larvae of O. volvulus were present in the skin of five cases, but were absent from four of the six which showed ‘ lichenification.’ It is, therefore, possible that the presence of the larvae in the skin may cause irrita- tion and lead to pathological changes. INCIDENCE. In order to gain some idea of the prevalence of this filarial infection in the Gold Coast, fifty men, taken at random, were examined at Accra. All the subjects were adults between the 469 ages of 25 and 45 years, who appeared to be in good health. The examinations were made between 9.45 and 10.15 a.m. on the 24th of October, 1922. From each man a small piece of skin, similar to those taken for skin-grafting by Reverdin’s method, was removed from the small of the back and placed in a tube containing about 2 c.c. of normal saline solution. The piece of skin was subsequently teased up Epidermis - - - - - - - Rete mucosum - - - - - Posterior end : ip : of Larva - - - - rs cae Sennen \ iy one *s a ann > . ~~ = Vic. 2.—Photo-micrograph of section of skin to show the position of the larvae. together with a drop or two of the saline solution from the bottom of the tube, the preparation fixed by heating, dried, and stained with haemalum. In no case was there obvious blood in the specimen. The result of this examination was that filarial larvae of the species here described were found in twenty-two of the men (equal to 44 per cent.). It may be noted, moreover, that larvae of 470 O. volvulus were found in seventeen (equal to 34 per cent.), and that in eight of these cases the other larva was also present. PERIODICITY. Ten of the men referred to above, in whom the larvae had been found, were re-examined two days later at about 9 p.m. No sensible difference suggesting periodicity in the prevalence of the larvae in the skin was observed.. DiaGNosis. The larva may be distinguished at a glance from that of O. volvulus, which also occurs commonly in the skin, by its slender body, crook-shaped posterior extremity, and blunt tail. In some respects it resembles the larva of Flavia demarquayi, but, apart from the fact that it apparently does not occur in the blood, it differs in that the tail is blunt, not sharply pointed, and that the column of nuclei extends practically to the tip of the tail. The larva from which it has to be distinguished most carefully is that of Ac. perstans, which also is sheathless and striated and has a stumpy tail to the tip of which the column of nuclei extends, and which occurs in the blood, but may also be found in small pieces of skin removed in the manner described. The descriptions of the larva of Ac. perstans which we have been able to find are somewhat meagre, and do not agree iu every respect. For example, Stephens (1916) gives the following measurements, length 160” to 2104, breadth 5 to 6m, nerve ring 34m, excretory pore 49m, genital pore 125#, and notes that smaller larvae occur measuring gomu to I10p by 4#; Rousseau (1019) gives, length 145m to 185m, breadth 3°54 to 5m, nerve ring 25 per cent., excretory pore 32 per cent., G1 cell 60 per cent., and anal pore 84 per cent.; and Johnston (1914) gives, length 83 to 170, nerve ring 232 per cent., excretory pore 32°9 per cent., G1 cell 62°6 per cent., and anal pore 83°5 per cent. In order to obtain comparable data, twenty larvae of Ac. perstans, fixed and stained in the same manner as the other larvae, were measured by us. In these specimens the length varied from 158m to 214m, average 170°4, breadth 2°75" to 5m, and the approximate positions of the nerve ring, excretory pore, G1 cell, and anal pore in a larva of the average length (1794) were respec- tively 22°5, 32°7, 62°3, and 81'1 per cent. of the length from the anterior extremity. The larva of Ac. ferstans is, therefore, shorter than the larva described in this paper, relatively stouter, and the nerve ring, the 471 Gi cell, and the anal pore are situated more anteriorly. When fixed in the manner described, it is, moreover, straight and not crook-shaped at its posterior extremity, and the column of nuclei at the anterior end is not reduced to a row of ten to twelve nuclei in single file. For the new parasite we propose the name Agamoflaria Streptocerca. REFERENCES Jounsron, J. E. L. (1914). Observations on Variations in Form of Microfilariae found in Man. Ann. Trop. Med. & Parasitol., Vol. VIII, pp. 73-79- Roussgau, L. (1919). Filariose au Cameroun. Bull. Soc. Path. Exot., Vol. XII, pp. 35-51- Srepuens, J. W. W. (1916). In ‘ The Animal Parasites of Man,’ p. 416. John Bale, Sons and Danielsson. London. THE UNIVERSITY PRESS OF LIVERPOOL Publications ys ah Liverpool School of Tropical Medicine Memoirs I-XXI, 1899-1906, contained reports of the numerous Expeditions of the School and other papers. Annals of Tropical Medicine and Parasitology. Vols. I-XVI, 1907-1922. Price £1 2s. 6d. per volume, unbound, post free. ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY Vol. XVI. No. 1. March 31, 1922 Undulant Fever in the Goat in Malta. By T. Zammit, C.M.G., M.D. Two Plates. Undulant Fever in the mye’ Military and Civilian Populations of Malta. By J. W. W. STEPHENS, M.D., F.R.S. . Alastrim ; or, Kaffir Milk Pox. By L. M. Moony, M.D., B.S., M.R.C.P. (Lond.) Five Plates. A New Species of Phlebotomus from Trinidad. By Professor R. NEWSTFAD, F.R.S. A New Tsetse-Fly from the South Cameroons. By Professor R. NEWSTEAD, F.R.S., and Miss ALWEN M. Evans, M.Sc. Notes on Australian Cestodes. Part IIJ. By P. A. MapLestoneE, D.S.O., M.B., Ch.B. Notes on Australian Cestodes. Part I1V. By P. A. MAapLeEsTone, D.S.O., M.B., Ch.B., and T. SOUTHWELL, A.R.C.Sc., F.Z.S. A Contribution to the Knowledge of the Bionomics of Sand-Flies. By Captain Jamrs WatersTon, R.A.M.C. (T.) One Plate. Human Intestinal Protozoa in Amazonas. By C. J. YounG, M.B., Ch.B. A Parasite Resembling .P/lasmodium falciparum in a Chimpanzee. By B. BLacklock, M.D., D.P.H. and S. ADLER, M.B., Ch.B. One Plate. ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY Vol. XVI. No. 2. July 20, 1922. The Signs of Filarial Disease. By B. BLacktock, M.D., D.P.H. Two Further Cases of Cardiac Aneurysm. By A. INGRAM, M.D., and J. W. S. Macrir, D.Sc., M.B. One Plate. Cestodes in the Collection of the Indian Museum. By T. SouTHWELIL, M.Sc., A.R.C.Sc. Cryptocotyle lingua (Creplin, 1825), Fischoeder, 1903, in a Dog in England. By P. A. MAPLESTONE, D.S.O., M.B., On the Genital Armature of the aC Mosquito. By J. W. S. Macrig, D.Sc., M.B., and A. INGRAM, M.D. Notes on Australian Cestodes. Part V. By P. A, MAPLESTONE, D.S.O., M,B., Cb.B., and T. SOUTHWELL, M.Sc., A.R.C.Sc. The Incidence of a Disease in Population Groups, the Number of People in which is known or unknown. By J. W. W. STEPHENS, M.D.,F.R.S. The Experimental Infestation of Physopsis africana. By F. G. Cawston, M.D., Cantab. Notes on Culicidae in Venezuela, with Descriptions of New Species. Part II. By Miss ALWEN M. Evans, M.Sc. One Pilate. Ancylostomes recorded from Sixty-seven Post-mortems performed in Amazonas. By R. M. Goxpon, M.D. ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY Vol. XVI. No. 3. October 18, 1922. The Susceptibility of the Individual to the Bites of Stegomyia calopus. By R. M. Gorpon, M.D. Tuberculosis in the Sudan, with Notes on a Case of Breast Tuberculosis in a Sudanese. By R. G. ARCHIBALD, D.S.O., M.D. One Plate. West African Ceratopogoninae. Part Il. By A. INGRAM, M.D., and J. W. S. Macrig, D.Sc., M.B. The Pathological Effects Produced by Strongyloides in a Chimpanzee. By B. BLACKLOocK, M.D., D.P.H., and S. ADLER, M.B., Ch.B. One Plate. Pulmonary Lesions in Dogs and Cats Naturally Infected with Nematodes. By B. BLACKLOocK, M.D., D.P.H., and S. ADLER, M.B., Ch.B. Ancylostomes in Animals in Freetown. By S. ADLER, M.B., Ch.B. The Occurrence of Ancylostomes Resembling Necator americanus amongst Domestic Pigs in Amazonas. By R. M. Gorpon, M.D. Parasites in Dogs and Cats in Amazonas. By R. M. Gorpon, M.D., and C. J. Youne, M.B. Chops A Noteon the Prevalence of Ceratopogonine Midges on the Windows of the Accra Laboratory during a Completed Year. By A. INGRam, M.D., and J. W. S. Macriz, D.Sc., M.B. Notes on Australian Cestodes. Part VI. By P. A. MapLestone. D.S.O., M.B., Ch.B. The Ascaris of Cattle. By J. W. S. Macrir, D.Sc., M.B. Mosquitoes Collected in the Mandaos Region of the Amazon. By R. M. Gorpon, M.D., and Miss ALWEN M. Evans, M.Sc. One Plate, Trypanosoma rhodesiense in a Case of Sleeping Sickness from the Sudan. By R. G. ARCHIBALD, D.S.O., M.D. ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY Vol. XVI. No. 4. December 30, 1922 An Uauenal Type of Nodular Leprosy in the Sudan. By R. G. ARCHIBALD, D,S.O., M.D. One Plate. Ancylostoma braziliense. By CLAYTON LANE. Intra-uterine Infection with Ancylostoma caninum in Dogs. By S. ADLER, M.B., Ch.B., and E, J. CLARK, M.B. Cestodes from Indian Birds with a Note on Ligula intestinalis. By T. SourTHWELL, M.Sc., AIRC ISG, HeZi5s A New Malaria Parasite of Man. By J. W. W. STEPHENS, M.D., F.R.S. One Plate. Notes on the Bionomics of Stegomyia calopus, Meigen, in Brazil. By C. J. Younc, M.B., Ch.B. The Occurrence of the Larvae of Onchocerca volvulus (Leuckart), 1893, in the Skin of Natives of the Gold Coast. By J. F. Corson. Two Plates. A Case of Sleeping Sickness (7. gambiense) Treated by ‘ Bayer 205.’ By J. W. W. STEPHENS, M.D., F.R.S., and WARRINGTON YORKE, M.D. Notes on the Bionomics of Stegomyia calopus, Meigen, in Brazil Part II. By R. M. Gorpon, M.D Observations on the Role of Cockroaches in Disease. By J. W. S. Macrik, D.Sc., M.B. The Occurrence of Xenopsylla astia, Roths., in West Africa. By Miss ALWEN M. Evans, M.Sc. Notes on a Case of Blackwater Fever. By ADAM PaTRICk, M.D. Case of Trypanosomiasis. By A. J. MACKENziIr, M.B., Ch.B. (Edin.) Observations on Onchocerca volvulus. By J. W. S. MACFIE, D.Sc., M.B., and J. F. Corson. A New Species of Filarial Larva Found in the Skin of Natives in the Gold Coast. By J. W. S. Macrie, D.Sc., M.B., and J. F. Corson. Bah vi a ¥ i . \ wid i $ ¢ ia F Q 1 vty at } . ‘ 8 14 j i ai i A; i \ { RC Annals of tropical medicine 960 and parasitology A55 Biological av & Medical A : ( Serials PLEASE DO NOT REMOVE CARDS OR SLIPS FROM THIS POCKET UNIVERSITY OF TORONTO LIBRARY STORAGE