|::i B 52745-1 v}m^-^"' iiiwAixT-^^y ;iF Tim ■ * . .0- ; ^--*i*^f TtlK oraiort/.— Preliminary Report on the Stndy of Cattle and Carahaos in the Philippine Islands. By James W Johling, M. D., Director of the Ir^ernm Laboratory. No. 5, 190$, Biolo(iicol Ljohorafory. — Trypanosoma and Trypanosomiasis, with Special Refer- ence to Surra in the Philippine Islands. By W. E. Mnsgrave, M. D., Acting Director Biological Laboratory, and Moses T. Clegg, Assistant Bacteriologist Biological Labora- tory. No. 6. 1903. — New or Noteworthy Plants, I. The American Element in the Philijtpine Flora. By Elmer D. Merrill, Botanist. (Issued .Tannary 20. 1901.) No. 7, 190$, iJfieniival /.((7>oraior//.~The Gntta Percha and Rubber of the Philippine Islands. By Penoyer L. Sherman, jr., l*h. D., Chemist, (^hemical Laboratory. No. 8, 190$.~A Dictionary of the Phmt Names of tlie Philii)pine Islands. By Elmer D, Merrill, liotanist. No. 9, 190$, Biological Laboratory. —A Report oti Hemorrhagic Septicamiia in Animals in the Philippine Islands. By l^aul (L WooUey. M. D., and J. W. .Jobling, M. D. No. 10, 190$, Biological J Aihoratory. —Two case<^ of a Peculiar Form of Hand Infection (Due to an Organism Resem]>ling the Koch-Weeks Bacillus). By John R. McDill, M. I)., and Wm. B. Wherry, M. D. No. 11, 1903, Biological hahoratory. — I^ntomological Division, Bulletin No. 1: Preliminary Bulletin on Insects of the Cacao, (Prepared Especially for the Bern e lit of Farmers.) By Charles S. Banks, Entomologist, Bureau of Government Laboratories. No. IP., 190$, Biological Laboratory. —Report on Some Pulmonary Lesions I'rodueed by the Bacillus of Hemorrhagic SepticTemia of Carahaos. By Paul (J. W^ooMey, M. D. No. 13, 190f^, LMological LAihoraiory. — A Fatal Infection by a Hitherto Undescribed Chromo- genic Bacterium: Bacillus aureus fa^tidus. By Maximilian Herzog, M. D. No. lU, 190fu — Scrum Laboratory: Texas Fever in the Philippine Islands and the Far East. By J. W. Jobling, M, I)., and Paul G. W^oolley. M. I). Biological Laboratory: Entom- ological Division. Bulletin No. 2. The Australian Tick (Boophilus anstralis Fuller) in the Phili]»pine Islands. By ('harles S. Banks, Entomologist. No. 15, 190!^, Biological and Serwin. Laljoratories.—liQpoTi on Bacillus Violaceus Manilce: A Pathogenic Micro-Grganism. By Paul G. Wool ley, M. D. No. 16, 190ft, Biological LAiboratory. —FTOteviivc Inoculation Against Asiatic Cholera: An Experinu'nlal Study. By Richard P. Strong, M. I). No. 11, 190/t.—^QW or Noteworthy Philippine Plants, II. By Elmer D. Merrill. Botanist. No. 18, 190U, Biological LMborat.ory.~~l. Amebas: Their Cultivation and Etiologic Signifi- cance. By W. E. Mnsgrave, M. D., and Moses T. Clegg. II. The Treatiuent of Intesti- nal Amebiasis (Amebic Dysentery) in the Tropics. By W'. E. Mnsgrave, M. D. No. 19, 190ft, Biological Labor atory.—i-^ome Observations on the Biology of the Cholera Spirillum. By W. B. Wherry, M. D. No. 20, 190ft.— Biological Laboratory: I. Does Latent or Dormant Plague Exist Where the Disease is Endemic? By Maximilian Herzog, M. I)., and Charles B. Hare. Serum Laboratory: 11. Broncho-Pneumonia of Cattle: Its Association with B. Bovisepticus. By Paul G. Woolley, M. D., and Walter Sorrell, D. V. S. HI. Pinto (Pafio Blanco). By Paul G. Woolley, M. 1). Chemical Laboratory: IV. Notes on Analysis of the Water from the Manila Water Supply. By Charles L. Bliss. Serum Laboratory: V. Frambeesia: Its Occurrence; in Natives in the Philippine Islands. By Paul G. W'oolley, M. D. No. 21, 19:)ff, Biological LMboratfjry.—iioiiie Questions Relating to the Virulence of Micro- organisms with Particular Reference to Their Immunizing Powers. By Richard P. Strong, M. D. (Coiitintied on third page of cover.) No. 31.— MAY, 1905 DEPARTMENT OF THE INTERIOR BUREAU OF GOVERNMENT LABORATORIES BIOLOGICAL LABORATORY I. NOTES ON A CASE OF H/EMATOGHYLURIA TOGETHER WITH SOME OBSERVATIONS ON THE MORPHOLOGY OF THE EMBRYO NEMATODE FILARIA NOCTURNA By Wm. B. Wherry, M. D., and John R. McDill, M. D. II. A SEARCH INTO THE NITRATE AND NITRITE CONTENT OF WITTE'S 'PEPTONE" WITH SPECIAL REFERENCE TO ITS INFLUENCE ON THE DEMONSTRATION OF THE INDOL AND CHOLERA-RED REACTIONS By William B. Wherry, M. D. MANILA BUREAU OF PUBLIC PRINTING 1905 31585 LETTER OF TRANSMITTAL Department of the Interior, Bureau of Government Laboratories, Office of the Superintendent of Laboratories, Manila, P. L , May 19, 1905. Sir: I have the honor to transmit herewith and to recommend for publication ^ ^I. Notes on a Case of Hsematochyluria, together with Some Observations on the Morphology of the Embryo Nematode Filaria Nocturna," by Dr. William B. Wherry, Bacteri- ologist, Biological Laboratory, and Dr. John R. McDill, Manila, P. I., and ^*II. A Search into the Nitrate and Nitrite Content of Witte's * Peptone,' with Special Reference to its Influence on the Demonstration of the Indol and Cholera-Red Reactions,'' by Dr. William B. Wherry, Bacteriologist, Biological Laboratory. Very respectfully, Richard P. Strong, Director Biological Laboratory, Acting Superintendent Government Laboratories. Hon. Dean C. Worcester, Secretary of the Interior, Manila, P I. 3 I. NOTES ON A CASE OF H^MATOCHYLURIA. TOGETHEK WITH SOME OBSERVATIONS ON THE MORPHOLOGY OF THE EMBRYO NEMATODE FILARIA NOCTURNA. By Wm. B. Wherry, M. B. , Bacteriologist, Biological Laboratory, and John R. McDill, M. D., Manila, P. I. The diagnosis of many cases of filariasis in which the adult parasites are inaccessible must depend upon the identification of the embryo nematodes. Most of the published pictures of Mlaria nocturna have been drawn apparently from stained preparations, and all the high-power photomicrographs we have seen, depict the parasite in a greatly degenerated condition and do not present the morphological details observed in a fresh preparation. The study of a case of hsematochyluria during the past six months and the accessibility of a Zeiss photomicrographic apparatus have given us the opportunity of presenting the accompanying illustrations together with an abstract of our notes on the case. ABSTRACT OF CLINICAL HISTORY. On May 30, 1904, Saya, a Japanese girl, 22 years old, came under our observation. She came to Manila four years ago from a village near Nagasaki. The patient was emaciated, pale, and weak and complained of passing bloody and milky urine and of attacks of abdominal pain referred chiefly to the right lumbar region. The abnormal urine was first noticed in August, 1903. This disappeared spontaneously in three months and she had no further trouble until the present attack. She claimed to have always drunk boiled water or tea and she gave a history of pre- vious good health. The patient was placed in the Manila Civil Hospital, where during the evening and at night she was pro- tected by a mosquito net. Upon admission she was passing a considerable amount of mill^y, 5 peach-colored urine, sometimes quite bloody, which upon cooling contained large and small clots of reddish and yellowish jelly-like material. These fibrinous clots were sometimes passed through the urethra and occasioned some pain. In the centrifugal precipitate a of number filaria-like organisms were found, but as none could be observed in the peripheral circulation during the day or at night, the patient w^as put on tonics and boric-acid bladder irrigations until the end of June, when urotropin, 1 gram three times a day, was given and the douches changed to bichloride of mercury 1-10,000, while morphine was administered hypodermatically for the pain. At 9 p. m. July 23 filaria, resembling those occurring in the urine, were found in the blood from a finger. The patient was kept in a bed, the foot of which was elevated, and received just enough food, without fats, and liquid to sustain life, and a "very weak solution of adrenalin chloride was injected into the bladder and allowed to remain. From August 1 to 15 the adrenalin was given by mouth, in doses of 10 to 15 drops of a 1-1,000 solution every four hours during the daytime, 40 to 50 drops per day. On August 15 this treatment w^as stopped. At this time some swelling of the right thigh developed but subsided after a few days. The patient remained in the elevated bed until the middle of October. On August 29 methylene blue, 0.12 gram every four hours, was given by mouth. This was stopped on September 4 on account of the occurrence of violent emesis. At the suggestion of Dr. W. E. Musgrave w^e attempted to "sensitize" the adult parasites by the administration of quinine followed by the exposure of the body of the patient, through the lumbar region, to the X-rays. She was given 80 to 90 grains of quinine sulphate during forty- eight hours, followed by X-ray exposures of five minutes, with the tube 18 inches away. Quinine having been administered daily, these exposures were performed from 2 to 3 p. m. on September 8, 10, 11, 12, 14, 16, 17, 18, and again, after cinchonizing as before, at 9 p. m. on September 28 and 30.^ ^Unfortunately we are unable to state the exact hardness of the X-ray bulbs. In order to obtain a clear radiograph of the bones of the pelvis with this apparatus an exposure of fifteen minutes with the bulb at a distance of 5 inches from the skin surface is necessary. Filarial embryos in a thin layer of blood, collected after cinchonizing, exposed to the rays for five minutes with the bulb 16 inches away are not killed, but they squirm about in a very excited manner. On October 2 the skin over the chest and abdomen became reddened and hot. A chill and left pleurisy developed. Para- centesis produced about 600 cubic centimeters of straw-colored fluid on October 8. Skin scarlet all over body. All this time the urine remained thick and bloody, but on October 10 became normal and has remained so. The temperature throughout, except during the attack of pleurisy, remained about normal — 97° to 99.4° F. in the morning and 98° to 99.4° F. in the evening. Until the pleuritic attack the patient had gainetl 25 per cent in weight and general appearance, and, although an evening tempera- ture of 1° to 3° F. persisted until October 22, she regained strength so rapidly that on October 29 and 30 the X-ray was again applied for fifteen minutes, after quinine, with the bulb 5 inches away. Although the patient has been at home and walking about for the past two months, her chyluria has not returned. The living embryos still persist in her blood, and hence it is altogether likely that the treatment had no effect upon the adult parasites. SPECIAL FEATURES OF THE CASE. (1) The urine. — The analyses of the urine have yielded the usual findings in such cases, excepting our failure to extract fatty matters in appreciable quantities. The bloody, milky urine never altered its appearance on prolonged shaking with ether, even after making it alkaline with sodium hydroxide, and the evaporation residue seemed to consist of other than fatty extractives, though in one instance a trace of fat was found by testing for gl3^cerine. Its milky appearance may have been partly due to the considerable number of leucocytes it contained. The amount of albumin varied between 0.33 and 0.6 per cent. For example, an analysis by Mr. C. L. Bliss, physiological chemist of the Bureau, on August 26, gave: Quantity, 675 cubic centimeters; specific gravity, 1,026; reaction acid but turning, kept at 30°; albumin, 0.33 per cent, average of five tests by Esbach' s method; fat, trace, by glycerine test. (2) The blood. — Four days after admission a blood count gave 3, 100,000 reds and 6,000 whites. The ana3mia almost disappeared as the general condition of the patient improved, during the period when she was at rest with the hips elevated. The excessive loss of blood did not continue for a long time and the ana3niia did not reach the grave character of such cases as Herrick (1) described as- due to repeated losses of blood from hemorrhoids. On July 3O5 the number of parasites per cubic centimeter in the patient's peripheral circulation was calculated. In order to obtain drops of blood of known volumCj the method of collecting it and estimating the number of parasites per cubic centimeter used by Lathrop and Pratt was employed. Three equal-sized drops of blood were taken every two hours, beginning at 10 a. m. and ending at 6 a. m. on the following day. The average number of parasites present was then determined by counting the stained fflaria on a mechanical stage. The following chart shows the rise and fall in the number of filaria per cubic centimeter present dur- ing the different times of the day and night: /6O0 t^f-r 76 6 /977» /^0O \ /Zoo \ i / 00 i \ 800 , \ ioo v \ 644 J^Oq V 4fS 2 QO d3 „,-^ 1 Chart showing theYlse and fall in the number of fllaria in the patient's peripheral circula- tion during different times of the day and night. The figures in the left-hand column arbitrarilv represent the number of Alaria per cubic centimeter of blood, while those within the ruled squares indicate the calculated number of filaria per cubic centimeter of blood. Strong^ (2) in 1901^ reported the first case of infection in the Philippine Islands with Filaria nocturna in a European who had resided in Iloilo for two years. A differential count of the leuco- cytes showed 3 per cent of eosinophiles. 9 In 1902, Calvert, (3) working in Manila on Filipino prisoners of war, found four cases of filariasis — ^lymphatic varices and hydrocele. No description of the parasites is given, but they were of the nocturnal variety and probably the embryos of Filaria bancrofti. By means of extensive blood counting in three cases he showed the presence of a decided eosinophilia, which w^as most marked at the time when the embryos were absent from the peripheral circulation. Trichinosis was excluded, but apparently no exami- nation was made for uncinaria. In our case not so many counts were made. The eosinophiles varied from 6 per cent (10 a. m.) to 12 per cent (12 p. m. ) and were most numerous in the peripheral circulation at the time when the embryos occurred in greatest numbers, as follows: July 28, 10 a. m. [Reds, 4,340,000; whites, 11,000.] Per cent. Polj^nuclear and transitional leucocytes 55 Eosinophiles 6 Basophiles „ „ „„^_ 4 Large mononuclears 22 Small mononuclears _. ._ 13 TotaL 100 One hundred leucocytes counted; 1 normoblast and no filaria seen. August 9, 8 p. m. [Reds, 4,290,000; whites, 6,000.] Differ enfifd. Poly nuclear and transitional leucocytes Eosinoi)hiles ^ Basophiles Large mononuclears- _^ Small mononuclear Total -.„__ 8 p. m. 12 p. m. Per cent. Per cent. 72 71.50 10.50 12 1.50 1.50 S.50 5 12. 50 10 Two hundred leucocytes counted. Number of filaria per cubic centimeter. [Calculated from the average number in two slides.] 8p. m „__„ ^_„ ._„_„ 489 12 p. m „ „ 1,079 10 On the morning of August 10' the patient was given an ounce of magnesium sulphate and her stool carefully examined for signs of intestinal parasites. Nothing but a few ova of Trichocephalus dispar was found. We believe that trichinosis can be excluded and are not aware that the eosinophilia can be influenced by the Trichocephalus dispar which the patient harbors/ According to Manson (6) the presence of blood in the urine in such cases is not due to the rupture of blood vessels' but to ''the formation of blood corpuscles in the lymph long retained in the varicose vessels." Our failure to alter the sanguineous character of the urine by the administration of adrenalin, locally and by the mouth, seems to support this idea. On the other hand, the development of anaemia and the presence of a few normoblasts in the peripheral circulation w^oidd seem to indicate that at least a portion of the loss occurred through capillaries torn during the rupture of dilated lymph vessels, as is suggested by Scheube (7). (3) The embryo nematode has been well described by Manson, and our own study has been greatly influenced by his excellent descriptions. A brief description made on July 30 may be inserted here: A fresh preparation was made at 10 p. m., ringed with vaseline and a filaria watched for some time. It underwent the usual movements of coiling, uncoiling, and sliding forward and back- ward within itl sheath. At about the junction of the middle and posterior thirds there could be seen an irregularly elongated viscus-hke organ, which seemed to be composed of a granular ^Bemlinger (4) (Constantinople) has recently noted an eosinophilia of 48 per cent in aj^ase of multiple infection with the Medina worm. He says: "Elle est Igalement k rapprocher de F Eosinophilia observee la Filaria Bangumm hominis^ la Mlarm Loa, la Filaria Imitis, dn Chien, etc. ' ' Manson (5) has expressed the belief that a large number of filarial embryos in the peripheral circulation indicates a multiple infection with adult parasites. Apparently there is no definite information as to the fate of the embryos, but Bancroft computes their life duration at a few months. To our knowledge they have not been observed in the dead condition in the blood of man except when killed by some form of medication, as in Scheube' s case, which was treated with picric-nitrate ot potash. It seems possible that they may accumulate in the blood, and, if so, large numbers would merely indicate that the case was of long standing. The grade of eosinophilia, together with an enumeration of the embryos by the method of Lathrop and Pratt, especially if supported by a post-mortem or post- operative search for the adult parasites, would throw light on this point. 11 tissue that was almost whitish by transmitted light. In about two hours the motions were reduced to very slight squirming movements. The outer contour of the filaria was clear cut, but within its lateral borders the serrated edges of the transverse fibers of the musculo-cutaneous layer could be traced. A little more than halfway between the anterior end and the viscusdike organ there was a refractile V-shaped papilla with its apex turned toward the lateral border of the filaria. With the one-twelfth oil immersion and compensation ocular No. 8, the head end was observed and at its extreme tip a notched retractile lip could be seen. Owing to the rapidity of the retractile movements the number of notches was indistinguishable. In addition there was a short, refractile, needle-like process, which was seen to be pro- jected and withdrawn. A few minutes later, when the movements had become slower, the lip, when retracted, showed at least three refractile tooth-like projections, and the needle-like spicule was seen to be projected at about the level of the middle tooth. (See Figs. B and C, PL II. ) By watching carefully and noticing what was revealed during the contractions and relaxations of the circular muscular fibers, three narrow, refractile, and sinuous duct-like threads could be traced backwards until they united with the anterior end of the viscus-like organ. (See fig. 2, PI. I, and Fig. B, PL II.) The exact manner in which they terminated anteriorly could not be made out. No particular structure could be distingxiished pos- terior to the viscus-like organ, excepting a refractile, V-shaped papilla, like the anterior one, and situated on the same side of the body at a place slightly posterior to a point halfway between the hind end of the viscus and the tip of the tail. The loose, transparent sheath could be seen projecting beyond the posterior but not beyond the anterior end of the parasite. In six hours ecdysis was not complete, the viscus-like organ had disappeared, and refractile granules began to make their appearance in the protoplasm. The parasite was measured just before granular degeneration set in and was found to be: Length, 0.31 millimeter; greatest breadth, 0.0075 millimeter; from anterior tip to anterior end of viscus, 153 /a; length of viscus, 49.5 /a; from posterior end of viscus to tip of tally 114.76 /*. (Zeiss one-twelfth ocular micrometer 3.) The average of four measurements is 0.327 by 0.0074 millimeter. 12 We have no new morphological details to add, excepting the three duct-like threads which connect the viscns-like organ with the head of the embryo. These must be looked for as soon as the motions of the parasite become slow enough to permit the use of an oil-immersion lens, for the granular degeneration, which sets in soon after motion ceases, obscures all finer details. (4) Photographing the embryo.— It is quite difficult to obtain good high-power photographs of the live filaria, and Mr. Martin's success followed only the most |)ersistent eflforts. The ray filter must be dispensed with and the photograph taken very soon after the embryo is exposed to the rays of the electric arc, as it undergoes rapid granular degeneration and its motions cease much sooner than when subjected to ordinary daylight. PREVALENCE OF THE DISEASE IN THE PHILIPPINE ISLANDS. There seems to be little positive information as to the extent to which the disease prevails in these Islands. Strong's case, as mentioned, had resided in Iloilo for two years. Calvert's cases came from northern and southern Luzon. Scheube. in his map showing the geographical distribution of filariasis, included this island but none of the southern islands of the group. We have not had time to go into this side of the subject thoroughly, but inquiry shows that physicians who have been in Manila for from ten to forty years have only rarely encountered cases of chy- luria or chylocele. Elephantiasis seems to be absent and filarial lymphangitis and varicose lymph glands may have been over- looked in the past. One other case of chyluria was seen in a Filipino this year by Dr. Bartels, but the patient left for the provinces before his blood could be examined. Several English physicians say they have seen cases of filariasis at Iloilo.^ It seems that the disease may be imported into localities where the conditions for its transmission are apparently unfavorable. Our patient has been living with four other Japanese during four, three, three, and tw^o years, respectively. An examination of these four and of a number of other Japanese^ living in their neighborhood was made late at night with negative results. This is rather surprising when one considers that Oulex fatiganSj the ^ Since the above was written Mr. Hare of this Laboratory has encountered one case of filarial lymph scrotum in a Filipino residing in Manila. The embryos were found in the peripheral circulation. — R. P. S. 13 mosquito which acts as a favorable intermediate host in many parts of the world, is one of the commonest species of mosquito found in Manila. However, it should be remembered that, notwithstanding the existence of some very strong presumptive evidence, the exact manner in which filariasis is transmitted is still an open question. The brilliant observations of Manson and Bancroft showing the metamorphosis of Filaria nocturna in the bodies of certain mosquitoes, and the further confirmation and extension of their views by the more recent work of Low (8) and James, (9) all tend to convince one that the disease is transmitted by the bite of certain mosquitoes.^ But the facts that a number of persons can live for years with a filariated patient, when apparently a favorable intermediate host is present throughout the year, without their acquiring the disease, as in our case, and the similar cases cited by Maitland, (10) and the '' relative immunity ^ ' of Europeans and others who are careful with regard to their food and drink, raise the old question whether Manson may not have been correct in his original assumption that the filaria escape from the mosquito to some watery medium and then gain entrance to their definitive host. REFERENCES CITED IN THE TEXT. (1) Herrick, Jas. B. Repeated Small Hemorrhages as a Cause of Severe Anaemia. Jour. Anier. Med. Assoc, Sept. 27, 1902. (2) Strong, R. P. Circulars on Tropical Diseases, No 1, 1901, Chief Surgeon's Office, Manila, P. I. Report of Surgeon- Oeneral, U. S. Army. Washington, 1901, p. 212. (3) Calvert, W. J. Eosinophilia in Filariasis. Johns Hopkins Hospital Btdletm, 1902, 13, 133. ^In the older literature on the transmission of filariasis, Cnkx ciliaris L. was named as the intermediate host, but in recent years our knowledge of the Culicidx has been greatly extended, and it has been show^n by Theobald (11) that C. ciliaris L. is identical with (J. pipiens, and further, that in all probability Hanson's original work in China and Bancroft's later work in Australia was not done on C. pipiens L. but on C fatigans Wied. (12). This widespread species is a voracious night feeder and occurs in large numbers in Manila. According to Low, (13) '4t is the chief spreader of filarial disease in the West Indies, acting as an intermediate host for F. nocturna. * * * It is inefii- cient for F. demarquoH.'' That the intermediate host is not restricted to one genus or species of mosquito is show^n by the feeding experiments of James, (9) in w^hich Anopheles rossii Giles and possibly a species of Cnlex--(Mex microannulatm Theob. and Stegomijia scutellaris Walk.— were shown to be suited for the metamorphosis of Filaria nocturna. 14 (4) Rbmlihgek, p. C, R. Soc. Biologie, 1904, 67, 76. (5) Makson, p. Brit Med. Jour,, Sept. 1, 1900, 536. (6) Manson, p. Tropical Dkemes, 1903, p. 581. (7) ScHBUBE, B. Die Krankheiten der Warmen Lander, 1904, in which a very complete bibliography will be found. (8) Low, G. 0. A Recent Observation on Filaria Nocturna in Culex; Probable Mode of Infection in Man. Brit. Med. Jour., June 16, 1900 1466. (9) James, S. P. On the Metamorphosis of the Filaria Sanguinis Hominis in Mosquitoes. Brit Med. Jour., Sept. 1, 1900, 533. (10) Maitland, J. Note on the Etiology of Filariasis. Brit. Med. Jour., Sept. 1, 1900, 637. (11) Theobald, F. V. Monograph Cididdm, 1901, 2, 136. (12) Theobald, F. V. Monograph CuHddm, 1903, 3, 226-26. (13) Low, G. 0. The Development of Filaria Nocturna in Different Species of Mosquitoes. Brit. Med. Jour., June 1, 1901, 1336. DESCRIPTION OF PLATES.' Plate I. Fig. 1. Filaria nocturna (X about S^O, double exposure), showing the general morphology and the viscus-like organ at the junction of the middle and posterior thirds of the parasite. Fig. 2. Head end of Filaria nocturna (X about 880). The sheath, the three duct-like threads connecting the anterior end of the viscus-like organ with the head end of the embryo, and the transverse striations of the musculo-cutaneous layer may be seen. II. (Figures redrawn by T. Espinosa, anatomical artist of the Labora- tory, from originals. ) Fig. A. Represents a dead filaria showing granular degeneration. Fig. B. Draw^n from a filaria just before granular degeneration set in. Proportions about correct as seen with the Zeiss one-twelfth oil immersion compensation ocular No. 8. Length, 0.330 millimeter; breadth, 0.00765 millimeter. The distance between the anatomical markings were as follows: A-B, 97.92 fi) B-C, 53:55 m; C-D, 6L20 m; D-E, 64.26 m; E-F, 53.55 m; total, 330.48 ju or 0.33 millimeter. Fig. C Head end of filaria showing retracted lips and spicule. * Photomicrographs by Chas. Martin, photographer of the Laboratory. 15 j-:'^i. ^ '^?n^- --. ^^ i") '''i w 111 1 ::y. -i ^(, -:i.: ,..:..........; ...y, .:,■■....... .,■ :.. : ,;..,.,:: j',;:; : .,„, '„ .:■.: ':::......,.:iJ^^Z:J^.Sy:il'^i-^i^*^'*^ ■ ■■■■■■ -""--^'M