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ANNALS OF [TROPICAL MEDICINE
AND PARASITOLOGY.

LOI iM. ADITOAT | IO 2
YOOAOTI2 oAtAT aUA

ieee NIVERSIDYS OR SLIVERPOOL

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.
Peomesoren NEWolhADY MSc.) )-P> F:Ris?) ALSy F.E‘S., Hon, F.R.H.S:
Proressorn WARRINGTON YORKE, M.D.
Prorsessor B. BLACKLOCK, M.D.

VOLUME XVII
(April 18, 1923, to December 21, 1923)

With Frontispiece, eighteen plates, eighty-seven figures
in text, two maps, and eight charts

LIVERPOOL:
THE UNIVERSITY PRESS OF LIVERPOOL LIMITED

LONDON:
HODDER AND STOUGHTON LIMITED

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ACES LIBRARY

re CONTENTS

Ye at ;
No. I. April 18, 1923
PAGE

onuson, J. FE.

A Case of Creeping Eruption in a European in the Gold Coast es us I
Macriz, J. W. S.

Depth, and the Larvae and Pupae of Stegomyta fasciata, F.... ft Le 5
Macrig, J. W. S.

A Note on the Action of Lithium Chloride on Mosquito Larvae - 03 9
ApLrr, S.

Malaria in Lacerta in Sierra Leone ... af aa Hd ip oi 13

Epwarps, F. W.
New and Old Observations on Ceratopogonine Midges Attacking other Insects 19

Ture, J. E. W.; and Van Oornt, G. J.

On some Strongylid Larvae in the Horse, especially those of Cylicostomum ... 31

KorLan, ALEXANDER.
Avian Cestodes from New Guinea. II. Cestodes from Casuariformes a. 47

KorLan, ALEXANDER.
Avian Cestodes from New Guinea. III. Cestodes from Galliformes ... ial 59

Curisropuers, S. R.

An Anopheles of the he! CINE ape AMAZINICUS, SP. ae from
South America ... . 71

Buacktock, B.
The Etiology of Blackwater Fever ... By oe se a2 Te he 79

Evans, Atwen M.
A New Species and a New Variety of Culex from the Belgian Congo ... AY 89

Orpen, R. W.
Report on Sleeping Sickness in Eket District, Southern Nigeria ese Se 93

Evans, Atwen M. :
Notes on Culicidae in Venezuela, with Descriptions of New Species ... Ot, Aish

CONTENTS

No. 2. July 12, 1923

PAGE
MaptestoneE, P. A.
A Revision of the Amphistomata of Mammals... pe ay a9: pee
Maptestong, P. A.
Malaria in Australia ... = a - ols re! ie a e213
Wenyon, C. M.
Coccidiosis of Cats and Dogs and the Status of the Jsospora of Man ... eee = t
Gorpon, R. M.
A Further Note on the Occurrence of Ancylostomes Resembling Necator
americanus amongst Domestic Pigs in Amazonas... at pe frenetic

Aptsr, $.; and Crarxg, E. J.
A Case of Acute Ascending Paralysis in a Chimpanzee ... ay ss is OD

CONTENTS

No. 3. October 13, 1923

PAGE
StepHens, J. W. W.
Studies in the Treatment of Malaria—XXXII. Summary.of Studies I—XXXI 303

Maptestong, P. A.; and Souruwett, T.
Notes on Australian Cestodes oe ee i Ae ee 7 eee 37

SouTHWELL, T.; and Apter, 5S.
A Note on Ophiotaenta punica (Cholodovski, 1908), La Rue, I9gII ae EE

Briacktock, B.
A Pyrrhocorid Bug Capable of Biting Man ex ee oP ee oreo

Heutr, Major-General Sir Parrick.
Typhus Fever in Greek Refugees ifr ee ee ys ae en SAg

STUART, GEORGE.
The Value of the Sachs-Georgi Reaction in the Serological Diagnosis of Syphilis 359

Evans, Atwen M.
Note on Aedinus amazonensis, Lutz oe del $a ae se 377

Gorvon, R. M.
The Treatment of Amoebic Dysentery_ ... ay ie ae HM. wersol

Senwyn-Crarke, P. S.; Le Fanu, G. H.; and Incram, A.

Relapsing Fever in the Gold Coast... at oe a : ay hoe 2350
Apter, S.
Carbon Tetrachloride in Filariasis ... ee ah rh i. sie aT 427

Waiter, R. O.
Yellow Fever in the Gold Coast : Its Endemic and Epidemic Character ... 431

MiscELLANEA af = ae a ee $3 oe es A rng 39

vil

CONTENTS

No. 4. December 21, 1923

PAGE
Briacktock, B.; and Tuompson, M. G.
A Study of the Tumbu-Fly, Cordylobia anthropophaga Griinberg, in Sierra
Leone. <. ra a i. i a ane gif A. tefl 443
Duke, H. Lynpuurst
The Transmission of 7. congolense by Glossina palpalis ... eas ie Pam Id
Newsteap, R.
On a New Species of Phlebotomus from Japan... ae an ans een 4
Newsteap, R.
Glossina ziemanni, Griinberg, a Synonym of Glossina palpalis, Sub-species
Fuscipes, Newstead abs sd or Bf *, a # wy 533
Apter, S.; and Cummines, FE. H. Taytor
Malignant Growths in Natives of Sierra Leone ... ae ie as wixy eb S
Wricut, W. Rees
A Preliminary Account of the Results of Surveys for Breeding-Places of
Mosquitoes in North Wales ... a Ae oo e. 7 RA ac
Gorpon, R. M.
The Frequency of Indicanuria ... Fe Mas 6 wel a 475 549
MiscELLANEA ae sa ae er ca aa at a7 Bs Pomel:

vill

THE LIBRARY
OF THE

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THE SCHOOL, OF TROPICGALSMEDICINE
BRUSSELS

A CASE OF CREEPING ERUPTION IN
A EUROPEAN IN THE GOLD COAST

Jape) erat ORSON
(Received for publication 14 November, 1922
PLATE |

This skin affection 1s stated by Crocker (1903) and by Castellani
and Chalmers (1919) to have been first described by Robert Lee in
1875; according to Roubaud (1914) the disease was observed in
Norway by Hoegh in 1869, while Abraham, in a review of a paper
by Knowles (1916), said that cases were recorded in Edinburgh
‘more than sixty years ago,’ 1.e., before 1856.

When a cause has been found, it has usually been a larva of a
fly of the family O6¢estridae, particularly Gastrophilus and
Hypoderma. Castellani and Chalmers (loc. cz¢.) state that larvae
of G. haemorrhoidalis and G. nasalis, of Oestromyia satyrus and of
H. bovis and H. lineata have been found. Looss said that larvae
of Aucylostoma duodenale in their passage through the skin could
cause it. Sakurane (1917) found a Ligula parasite in a swelling of
the skin, and suggested that the parasite of creeping eruption is of
this nature. Ikegami (1919) removed from a case of this disease a
young worm, said to have been probably Lchinorhynchus sphaero-
cephalus, but a structure in it lke an alimentary canal suggested
Gnathostoma. Tamura (1919) removed a male Gurathostoma
resembling G. szamense.

The disease is reported to have occurred in Ireland, Scotland,
the Shetland Isles, Norway, Sweden, Denmark, Russia and Siberia,
Bulgaria, Arabia, Sumatra, China, Japan, the United States of
America, Brazil and West Africa (Senegal, Sierra Leone, Liberia,
Togoland, Nigeria and the Cameroons).

The form of the disease occurring in Senegal, called locally
Oerbiss or Larbish, and for which no cause has been found, is
considered by Roubaud to be of different aetiology from cases due
to myiasis.

The following case showed a close resemblance clinically to the
description of Oerbiss given by Roubaud.

Mr. G., British, living at Seccondee, Gold Coast, noticed, about
the 18th of June, 1922, a small itching spot on the ball of the left
thumb, and thought that it was probably due to a bite of some
insect. A few days aiterwards he noticed that the spot had become
a line, and by the 26th of June there was a curved, raised, blister-
like line about three-quarter inch long and one-sixteenth inch in
diameter. Itching and a burning sensation were considerable,
especially at night. On the 6th of July the appearance was as shown
in Plate I. From then onwards until the beginning of October,
when opportunities of observing the case ceased, the track
progressed irregularly and intermittently round and along the
thumb to near the tip. No parasite was found; microscopic
examinations of serum and blood taken from various parts of the
track and attempts at culture in broth and on agar and blood serum
were without result.

No serious attempt to cure the disease was made; an ointment
of sulphur and ammoniated mercury was used by the patient, who
also opened the tracks from time to time and rubbed in tincture
of iodine with apparent temporary benefit.

REFERENCES

ABRAHAM, P. S, (1882). Med. Press G Cire., April 12th. See Trop. Dis. Bull., Vol. VII,
No. 4, September, 1916, review of Knowles’s paper (see below).
(1897). Trans. Derm. Soc. G. B. & I., Vol. 111. See Crocker’s Diseases of the Skin,
3rd ed., 1903, p. 1309.
CasTELLANI and CHALMeERs (1919). A Manual of Tropical Medicine, 3rd ed., 1919. London:
Bailliere, Tindall and Cox. ‘The following are some of the references given :—
Biancuarp (1901). Arch. de Parasitologie.
Braun-Seirert (1908). Die tierischen Parasiten des Menschen.
Hutcurns (1908). Fournal of Cutaneous Diseases.
Crocker, H. R. (1903). Diseases of the Skin, 3rd ed., 1903. Lond.: H. K. Lewis. The
following references are given :—
Crocker (1896). Folio Atlas of Diseases of the Skin, plate XCIII, figs. 2 and 3.
Clin. Soc. Trans., Vol. VIII (1875), p. 443 Vol. XVII, p. 75.
NeuMANN (1896). Arch. f. Derm. u. Syph., Vol. XXXIV, Heft. 1, p. 905. Abs.
Brit. Four. Dermat., Vol. VIII, p. 145, of Russian papers.
SAMSON-HIMMELSTJERNE (1898). Ein Hautmaulwurf, with Plate of case, and
woodcut of Gastrophilus larva. Short abs. Amer. ‘four. Cut. and Gen. Ur. Dis.,
Vol. XVI, p. 297.
SmitH, WatTeER (1881). Rep. of Int. Med. Cong., Lond., with partial bibliography
and substance of McCalman’s case.
Dusreuitn, W. (1894). Les Diptéres Cuticoles chez 'homme. Arch. de Méd.
Exp., No. 2, with references.
Pucrra, Hrpexazu (1919). A Case of Creeping Disease. Hifu-Ka oyobi Hinyoki-Ka Zasshi
(Fl. Derm. & Urol.), No. 10. Review in Trop. Dis. Bull., Vol. XVUI, No. 2, Aug. 1921.

FUireporn, F., and pa Rocna Lima, H. (1919). Ueber Larbisch und Woloss-jatik (Hautmaul-
wurf). Arch. f. Schiffs u. Trop. Hyg., Vol. XXIII, No. 13. Rev. in Trop. Dis. Bull.,
Vol. XV, No. 3, Mar. 1920.
Gray, C. P. (1917). Larva migrans on the Mexican Border. New York Med. ¥l., Vol. CVI,
No. 1. Rev. in Trop. Dis. Bull., Vol. X, No. 4, Oct., 1917.
Horta, Parrerras (1917). Um caso de ‘larva migrans’ com varias galerias. Brazil Medico,
Vol. XXXI, No. 20. Rev. in Trop. Dis. Bull., Vol. X, No. 4, Oct., 1917.
IkrGami, Yutaka (1919). On the Cause of Creeping Disease. Hifu-Ka oyobi Hinyoki-Ka
Zasshi (Fl. Derm. & Urol.), No. 10. Rev. in Trop. Dis. Bull., Vol. XVII, No. 2, Aug.,
1921.
Kirey-SmitH, J. L. (igts). Extensive Creeping Eruption. New York Med. Fl., Vol. CI,
No. 11. Rey. in Trop. Dis. Bull., Vol. VI, No. 3, Sep., 1915.
(1916). Ausedehnte Creeping Eruption. Dermat. Woch., Vol. LXII, No. 11. Trop.
Dis. Bull., Vol. 1X, No. 9, June, 1917, list of references.
KraupeR, J. V., and Grrenzaum, S. S. (1921). Creeping Eruption (Larva migrans). Report
of a Case contracted at Wildwood, New Jersey. Arch. Derm. and Syph., Vol. II,
No. 4, Part 1. Rev. in Lancet, April 23, 1921, and in Trop. Dis. Bull., Vol. XIX,
No. 2, Feb., 1922.
Know ss, F. C. (1916). Creeping Eruption of the Skin. /. Amer. Med. Assoc., Vol. LXVI,
No. 3. Rev. in Trop. Dis. Bull., Vol. VIII, No. 4, Sep., 1916.
Macrir, J. W.S. (1918). A Case of Larbish or Oerbiss observed in Northern Nigeria. ¥/. Trop.
Med. & Hyg., Vol. XVI, No. 3.
Murakaml, F. (1917). ‘ Creeping Disease’: a Linear Circinate Disease of the Skin. Hifuza
Hitsumyokwa Zasshi (Fapan. Zeitsch. f. Derm. u. Ur.), Vol. XVII, No. 5. Rev. in Trop.
Dis. Bull., Vol. XI, No. 6, June, 1918.
Parnam, J. C. (1916). Creeping Eruption. Report of a Case. U.S. Nav. Med. Bull., Vol. X,
No. rt. Rev. in Trop. Dis. Bull., Vol. VU, No. 4, Sep., 1916.
Pawrovsky, E. N., and Stern, E.R.(1922.) Nouveau Cas de Creeping Disease (M/yiasts linearis)
en Russie. Bull. Soc. Path. Exot., Vol. XV, No.7. The following references are given :—
Cuotopkovsky, M. (1908). Ueber Gastrophilus-larven in der Menschenhaut. Zool.
Anz., Vol. XXXIII, p. 409.
LENGLET ET Desanay (1904). Un cas de larva migrans. Ann. de Derm. et de Syph.,
4¢ série, Vol. V.
PortscHINsky, J. A. (1907). Die Grosse Magenbremse des Pferdes (Gastropbilus
intestinalis). Saint-Petersbourg.
Rovusaup, E. (1914). Les Producteurs de Myiases et Agents Simulaires chez l’homme et les
animaux. Paris: Emile Larose. The following are some of the references given :—
Boas (1907). Monat. prakt. Derm. 44, quoted by Cholodkovsky. Zool. Anz., Vol.
XXXIII, Leipsig, 1908). .
BERENGER-FERAuD (1875). Traité clinique des maladies des Européens au Sénégal.
Fixiezorn (1908). Beibefte Arch. f. Schiffs u. Trop. Hyg., 6.
Horcu. Quoted by Marbitz in Bethefte Arch. f. Sch. u. Trop. Hyg., 6, 1908.
Kutz (1913). Arch. f. Sch. u. Trop. Hyg., Vol. XVII.
Looss (1905). Mense Handbuch d. Tropenkrankheiten, Leipsig, p. 13; new ed.,
1914, p. 406. Also, The Anatomy and Life History of Agchylostoma duodenale
Dub. Rec. of School of Med., Cairo, 1911, Vol. IV.
Mepenszacu (1910). Arch. f. Schiffs u. Trop. Hyg., Vol. XIV.
Ovoreo ve Morass (1913). Norte Medico, Ceara.
ScHOEGEN (1906). Entom. Tijsk., Vol. VII, p. 171.
Topsent (1901). Arch. Parasitol., Vol. IV.
Watpvon (1903). Arch. f. Schiffs u. Trop. Hyg., Vol. XIII.
Roveaup, E. (1914). Le Larbish ou Oerbiss des Ouloffs, pseudo-myiase rampante sous-
cutanée du Sénégal. Bull. Soc. Path. Exot., Vol. VII.
SakurANE, K. (1917). Creeping Disease and its possible relation to Ligula Infection. Hifucwa
Hitsumyokwa Zasshi (Fapan Zeitsch. f. Derm. u. Urol.), Vol. XVII, No. 5. Rey. in
Trop. Dis. Bull., Vol. XI, No. 6, June, 1918.
Tamura, Haruxicut (1919). On Creeping Disease due to ? Gnathostoma. Hifu-Ka oyobi
Hinyoki-Ka Zasshi (Fl. Derm. & Urol.), Nos. to and 11. Rev. in Trop. Dis. Bull.,
Vol. XVIII, No. 2, Aug., 1921.
(1921). On Creeping Disease. Brit. Fl. Derm. & Syph., Vol. XXXIII, Nos. 3 and 4.
Rey. in Trop. Dis. Bull., Vol. XIX, No. 2, Feb., 1922. This paper is stated in the
review to contain a full review of the literature.

EXPLANATION OF PLATE J

Case of Creeping Eruption.

Annals Trop. Med. @& Parasitol., Vol. XVII PRATESL

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DEPTH, AND “THE LARVAE AND
PUPAE OF STEGOMYIA FASCIATA, F.

J. W, 5S. MACFIE
(Received for publication 14 November, 1922)

The larvae of Stegomyia fasciata, as is well known, are usually
found in small collections of water in domestic utensils, old tins,
rot-holes in trees, calabashes, rock pools, etc.; they are to a large
extent bottom- and side-feeders, and are capable of remaining, and
in fact frequently do remain, completely submerged for very consider-
able lengths of time. In many tropical countries where S. fasczata
is prevalent, water is stored in tanks of some size and depth. Even
if efficiently screened, larvae may gain access to these tanks by being
washed in with rainwater as eggs or young larvae, and it 1s a matter
of some interest to know if, supposing they were introduced in this
way, the larvae of S. fasczata would be able to thrive or would be
likely to escape by being drawn off with water from a tap situated
near the bottom.

Iyengar (1920) observed at Calcutta that larvae of S. fasczata,
which are found there in almost all domestic situations in which
larvae of Anopheles stephenst are found, are, however, rarely
encountered with them in wells. He accounts for this difference in
habit by the fact that whereas the larvae of A. stephensi are provided
with hooks at the ends of the dorsal hairs on the ninth abdominal
segment by means of which they cling to the sides, the larvae of
S. fasciata lack these hooks, and, therefore, he assumes, were they
to frequent wells would have to go to the bottom if the water was
disturbed. ‘It is likely,’ he thinks, ‘that mosquito larvae, being
air-breathing organisms, cannot ordinarily stand much pressure at
the depth of a well. Stegomyia is a bottom- and _side-feeder ;
therefore it has to go to the bottom of its breeding-place, unlike
A. stephensi, which feeds on the surface. These facts explain why

Stegomyia has rarely been found in waters which are over three feet
deep.’

These statements set us wondering if it was, indeed, the case that
larvae of S. fasczata could not withstand the pressure of more than
about three feet of water, and if disturbed must inevitably go to
the bottom. The following experiments were carried out to ascertain
the facts.

The apparatus used consisted simply of a wide-mouthed bottle
with sloping shoulders, connected by a stout piece of rubber tubing
with a length of wide-bore glass tubing. The tube and the bottle
were set up vertically, the one above the other, and securely clamped.
When required, additional lengths of tubing were added at the top
with short rubber connexions.

In such a system larvae of S. fasczata lived apparently at ease,
and after a day or two congregated at the top, mostly in the first
foot, a few in the second, and only stray individuals at greater
depths. The successive stages observed in an actual experiment are
shown in the Table. As will be seen, the larvae, which at first were

| |
Day ri i] I 2 | 3 4 5 6
ist foot... ae ++ + +e +--+ +--+ +--+
(Mostly at | (Nearly all | (Nearly all
| the top) at the top) | at the top)
2nd foot ... rs -- | -|- 4--- + — 5 I
STOIGOl, sei oF -|- | -|- + — fe) I | fe)
4th foot ... ey -+|- | +— — I fe) O | fe)
sth foot... - -|- |} +-—— | fe) O ° Oo
6th foot... sat 4- + —— fo) e) ° oO
7th foot ... ve + +- | Oo O ° Oo

+--+ = many Larvae. -+- = several Laryac. + — = few Larvae.

-+-—— = very few Larvae. Tyas pronies five Larvae,

distributed throughout the tube, collected rather slowly at the top,
so that after three days almost all of them were in the first foot of
the column of water, and the majority at any particular moment
actually at the surface. During this process of settling the habits
of the larvae changed, bottom feeding being discontinued.

If then the tube was shaken or tapped, the larvae left the surface
and wriggled down in the usual manner. They did not, however,

sink to the bottom ; indeed, most of them descended only a few
inches and very few more than one foot. Their descent was not
passive, but was effected by active wriggling movements, and when
these ceased they immediately began to float upwards towards the
surface. Under ordinary circumstances, if disturbed the larvae
wriggled downwards a few inches, ceased wriggling and floated
upwards a short distance, and then recommenced active wriggling,
this time towards the surface. They did not attempt to cling to
the side of the tube. It is clear, therefore, that the larvae of
S. fasciata when disturbed do not necessarily go to the bottom.

As the result of a single tap on the tube, it occasionally happened
that one or two larvae descended to greater depths, such as two and
a half feet or even three and a half feet. Larvae were also some-
times observed to descend voluntarily as much as five feet, and
once one was found browsing on the side of the tube at a depth of
6 feet. By repeated tapping on the tube the larvae, could be urged
to descend even deeper, eight feet at least. They did not appear
to be at all incommoded by the pressure of the column of water,
and when the tapping ceased wriggled back to the surface.
Sometimes they rested on the bottom for a short time before starting
the upward journey. It took one larva six minutes to regain the
surface after descending seven feet.

In one experiment the system, consisting of the bottle and a
long glass tube of wide bore of a total length of seven feet, was left
standing until a copious growth of green algae had formed over
the bottorn, from which small bubbles of gas arose in sunlight and
presumably kept the water oxygenated. In this system larvae of
S. fasciata throve better than they did when no algae were present,
and were more frequently seen at greater depths; indeed, both
young and older larvae, but especially the former, were often seen
browsing actually on the bottom. The pressure of the seven-foot
column of water above them appeared to have no injurious effect
whatsoever.

The pupae of S. fasciata, however, are not able to descend
unharmed to such great depths as the larvae. As the result of a
single tap on the tube, they usually descended only an inch or two
and then floated passively back to the surface. By repeated tapping
they could be induced to descend considerably further, but beyond

a certain depth (which in our experiments appeared to be about
three feet to three and a half feet) they showed an unquestionable
anxiety to return to the surface, ceasing to respond readily to
disturbances, such as tapping or shaking, even when violently
applied, descending further only very reluctantly, and sometimes
refusing to move at all or actually ascending in spite of everything.
In one experiment, by means of repeated tapping and shaking, a
pupa was driven down to the bottom, a distance of seven feet.
From this position it struggled upwards, evidently with increasing
difficulty, for a distance of a little more than four feet. At about
this level it managed to maintain itself for several minutes, now
jerking itself up an inch or so, now sinking an inch or so, and then
began to lose ground, at first slowly, then more quickly, and
eventually sank to the bottom. Another pupa was similarly induced
to descend five feet, but it managed to regain the surface. The
inability of pupae to descend without ill-effects to such great depths
as the larvae appeared to be dependent on their diminished
buoyancy at such depths, which caused them to begin to sink the
moment active movement was arrested. This fact should be
correlated ‘with the imperative need of pupae of access to air, for
the strenuous efforts exerted in struggling upwards from an
unaccustomed depth no doubt accelerated the exhaustion of the
supply of air in their tracheal tubes.

REFERENCE

Tyencar, M. O. TrruNARAYANA (1920). Preliminary Report of a Malaria Survey of Calcutta
and Environs. Special Indian Science Congress Number, 1920, of The Indian Fournal
of Medical Research, pp. 15-16.

A NOTE ON THE ACTION OF LITHIUM
CHLORIDE ON MOSQUITO LARVAE

1S a We Cl OTD
(Received for publication 14 November, 1922)

It is well known that larvae of certain mosquitoes, e.g.,
Stegomyta fasciata, are intolerant of common salt. Other chlorides
act similarly, some of them very powerfully, as is shown in Table I,

Tasce I.

The number of hours required to kill all larvae of Stegomyia fasciata in various soloutions of

which summarises a series: of preliminary experiments on the action
of these salts carried out by Mr. R. Simmons, which I am permitted
by him to quote. Additional experiments were made subsequently
with lithium chloride and Séegomyia fasciata. In one of these, five
larvae and one pupa were placed in a 1'2 per cent. solution of LiCl;
within four hours all the larvae were dead, but the pupa appeared
to be unaffected. In three others, twenty-seven larvae were placed
in a 0°3 per cent. solution of LiCl in the afternoon; all were dead by
next morning, that is within sixteen or seventeen hours.

During the experiments it was noted that lithium chloride not
only killed the larvae of Stegomyia fasciata, but also produced a
peculiar effect on them, causing them to writhe about at the bottom
of the jars, apparently unable to rise to the surface, and to become
entangled with one-another, usually by the mouth brushes. These
effects were observed even in the weakest solutions used.

As lithium chloride appeared to have a very powerful effect on
the larvae, further experiments were carried out to determine the
limits of the injurious action.

Culex fatigans. The larvae were placed in glass jars (five in
each) containing 100 c.c of the lithium chloride solution. The jars
were covered with glass plates, stood on the laboratory bench, and
The
solutions used were 0°3, 0°15, 0°00, 0°03, and 0-015 LiCl per cent.
The results are summarised in Table II.

examined morning and afternoon at about 9 a.m. and 5 p.m.

SAREE Ube

The effect of solutions of Lithium chloride on the larvae of Culex fatigans.

Exactly similar experiments were carried out with larvae of
Stegomyia fasciata and Anopheles costalis. Without entering into
details, it may be said that the results also were similar, all, or
practically all, the larvae dying in the o'3 and o'15 per cent.
solutions within twenty-four hours, and in the 0°06 per cent. solution

within two or three days. In the 0°03 and o'o15 per cent. solutions
the larvae, especially when young, were also affected: in an experi-
ment with almost fully grown larvae of S. fasczata, for example, only
two out of ten completed their development in the former solution,
and two out of seven in the latter, whereas in the control jar no
casualties occurred. In the case of S. fasciata, entanglement of the
larvae by their mouth brushes and other setae was repeatedly, but
not invariably, observed.

It is worthy of note that the larvae of Mansonioides africanus,
which live attached to the roots of the water-weed Pistia stratiotes,
do not escape the action of lithium chloride. A small plant of Pzstza
stratiotes with larvae attached to it was placed one afternoon in a
jar containing 100 c.c. of a 0°3 per cent. solution. By the next
morning, that is within eighteen hours, all the larvae had left the
roots of the plant and were dead.

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MALARIA IN CHIMPANZEES IN SIERRA
LEONE

S. ADLER
(Received for publication 15 November, 1922)
PLATES II AND III

Reichenow (1920) working in the Cameroons, found parasites
indistinguishable from human malaria parasites in the blood of
gorillas and chimpanzees. Of eight chimpanzees examined six were
found to be infected, one with Plasmodium vivax ? forms (gameto-
cytes only), two with Plasmodium falciparum ‘forms (crescents
only), one with P. falctparum and P. vivax forms, and two with
P. falciparum and P. vivax forms together with P. malariae forms.

Reichenow found that infections were heaviest in young animals,
and suggested that resistance is acquired after attacks in early life.

Blacklock and Adler (1922), of the Liverpool School of Tropical
Medicine, described a parasite resembling Plasmodium falciparum
in a chimpanzee, and forms resembling P. vivax and P. malariae
also occurred, but the only form of gametocyte found was the
crescent.

I have recently examined thirteen additional chimpanzees, six of
which were caught near Pendembu, and six near Biama, in the
Sierra Leone Protectorate, and one from an unknown locality.

Of these thirteen animals, two were found to be infected with
parasites indistinguishable from P. falciparum. The infected cells
were not enlarged or pale, and many of the delicate rings showed
two bars of chromatin. In both cases crescents were found, but only
after prolonged search, resernbling in this respect human infections
with P. falciparum in West Africa. Parasites resembling simple
tertian or quartan forms were not found.

CASE I. Captured near Blama. The animal was emaciated
and weak. A blood examination on 8th September, 1922, showed
numerous rings and a few crescents. On oth September, 1922,
quinine hydrochloride, 0°5 grains, was administered intramuscularly ;
rings were present in the blood until 14th September, 1922, but
crescents persisted until the animal’s death on 2nd October, 1922.

. 7

The animal’s condition showed no marked improvement after the
disappearance of rings from the peripheral blood; its appetite was
poor and it often passed loose stools containing a large amount of
fat globules. Death occurred after an attack of enteritis, which was
apparently caused by an invasion of Oxyuris sp., of which large
numbers (all immature) were passed in the animal’s stool.

Post-mortem, malaria pigment was found in the spleen, liver,
and bone marrow, and crescents, in small numbers, in the bone
marrow. The liver showed fatty changes. Enormous numbers of
immature Oxyurzs were found in the large intestine.

25th September, 1922. Advantage was taken of the fact that
rings had not been seen in the blood for eleven days, and that
crescents still persisted, to test the theory of parthenogenesis.
o°4 c.c. of the animal’s blood were injected into another chimpanzee
in which malaria parasites had never been found since it first came
under observation on 4th September, 1922.

The injected animal was observed till the 11th November, 1922,
but parasites were not found in the peripheral blood.

CASE II. Captured near Pendembu. The animal was extremely
emaciated. On 12th September, 1922, rings and crescents were found,
but the infection gradually disappeared without treatment, and on
the 19th September, 1922, the blood became negative. The animal’s
condition gradually became worse, it took very little food, the stools
were loose and always contained fat globules.

On 30th October, 1922, the blood examination again showed a
few ring-form parasites.

The animal died on 30th October, 1922. Post-mortem
pigment was found in the spleen and bone marrow, and a small
number of schizonts in the spleen; no crescents were found. The
liver was pale, and on section showed extreme fatty degeneration
and infiltration, the majority of the liver cells being destroyed.

It is interesting to note that both animals were young (under two
years). Older animals, including one old adult, were negative.
This supports Reichenow’s suggestion that in chimpanzees, as in
natives, resistarice 1s acquired after attacks in early life.

THE RELATIONSHIP OF MALARIA IN CHIMPANZEES TO
HUMAN MALARIA IN SIERRA LEONE

Although the malaria parasite in the chimpanzee in Sierra Leone
is morphologically indistinguishable from P. falciparum, there is as
yet no evidence that it is this species.

Mesnil (1920) failed to infect a chimpanzee by intravenous
injections of human blood infected with P. falczparum. He also
failed to infect the same chimpanzee by the bite of Anophelines with
sporozoites of Plasmodium falciparum in their salivary glands.

Blacklock and Adler (1922) failed to infect :—

(1) Two Europeans by intravenous and subcutaneous injections
of heavily infected blood from a chimpanzee.

(2) A chimpanzee by an injection of 3 c.c. of blood heavily
infected with P. falczparum from a patient during his first attack of
malaria, which he acquired in Sierra Leone.

(3) Anopheles costalis by feeding on a chimpanzee; but it should
be noted that crescents were scanty in the animal’s blood.

The existence of a relationship between human malaria and
malaria in chimpanzees cannot be conclusively proved or disproved,
until the insect vector of the latter be discovered and experiments
with the infective vector carried out on human beings.

My best thanks are due to Mr. W. Addison, Provincial
Commissioner of Kennema, and Mr. N. C. Hollins, District
Commissioner of Pendembu, through whose kindness I obtained a
number of chimpanzees.

SUMMARY AND CONCLUSIONS

Thirteen chimpanzees were examined for malaria in Sierra Leone.

Two young animals were found to be infected with a parasite
indistinguishable from P. falciparuzz.

Older animals were negative, and resistance following attacks in
early life is, therefore, suggested.

Blood trom one chimpanzee containing only crescents failed to
infect another chimpanzee.

Both infected animals on post-mortem examination showed fatty
changes in the liver.

REFERENCES

Bracxiock and ApLeR (1922). A Parasite resembling Plasmodium falciparum in a chimpanzee.
Ann. Trop. Med. & Parasit., Vol. XVI, No. 1, pp. 99-106.

Mesnit, F., and Rousaup, E. (1920).° Essais d’inoculation du paludisme au chimpanzé. 4nn.
Inst. Past., Vol. XXXIV, No. 7, pp. 467-479.

REICHENOW, “E. (1920). Ueber das Vorkommen der Malaria Parasiten des Menschen bei den
Afrikanischen Menschenaffen. Centralblatt f. Bakteriologie. Abt. Originale.
Vol. LXXXV, No. 3, pp. 207-216.

EXPLANATION OF PLATE II
Malaria Parasites.

Figs. 1 to 17. Ring forms.

Figs. 18 to 19. Crescents.

Annals Trop. Med. & Parasitol., Vol. XVII

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Fig. 1. Micro-photograph of liver (Case II), showing fatty
degeneration and infiltration. x 250.

lig. 2. On the right, young chimpanzee with malaria, showing
emaciation. (Note absence of paunch.)
On the left, healthy young animal.

PLATE Ill

Annals Trop. Med. & Parasitol., Vol. XVII

Fic. 2

C. Tinling & Co., Ltd., Imp.

FHE (iBRARY
OF THE
NWIVERSITY OF ILLINOIS

NEW AND OLD OBSERVATIONS
ON CERATOPOGONINE MIDGES
PPA CKINGOT HE RIM INSEGYTS

F. W.. EDWARDS

(Published by permission of the Trustees of the British
Museunt)

(Received for publication 29 November, 1922)

During the last two decades a fairly extensive literature has
been published in regard to the attacks made by Ceratopogonine
midges upon other insects. Several cases were placed on record by
Knab (1914), who also reviewed the literature published previously
to this date. More recently the facts have been reviewed by
Peyerimhoff (1917) and Kieffer (1922), the former author adding
some very interesting observations of his own. There are, however,
one or two additional and very interesting cases which have been
overlooked by all the above-named writers, and also some further
unpublished instances which have come to my notice. It may,
therefore, be worth while, in recording the fresh cases, to review
again the literature of the subject.

The attacks made by midges upon other insects fall under two
main heads :—

(1) Cases of predacity, where the midges attack other adult
insects of approximately their own size, or not much larger, and
kill them by puncturing their skin and sucking them dry. A good
many instances of this have been noted, and I have summarised them
in a recent paper (1920).

The following list gives the names of these species and of their

victims : —
PREDATOR VicTIM
NEUROPTERA
Palpomyia flavipes, (Meg.) Ephemerid (Baetis sp.)
» Sp. Perlid
CHIRONOMIDAE
Bezzia annulipes (Mg.) Tanytarsus sylvaticus, v.d. Wulp.
Probezzia multiannulata (Strobl.) Culicoides circumscriptus, Kieffer.
Probezzia ? signata (Mg.) Culicoides pulicarts (L.)
Stilobezzia gracilis (Hal.) Cricotopus pulchripes, Verr.
- A Orthocladius sp.
ie 4s T anytarsus, 2 spp.
of ne Tanypus binotatus, Mg.
Serromyia femorata (F.) Cricotopus pulchripes, Verr.
. e Bezzia ornata (Mg.)
ne + Serromyia femorata (F.) 3
- re Trichocladius sp.
+Ceratopogon candidatus, Winn. Trichocladius sp.
Ceratopogon lacteipennis, Zett. Camptocladius ? gracilis, Goet.
ts a Culicoides arcuatus (Winn.)
f Ceratopogon lacte1pennis Zett., 3

In addition, Kieffer (1922) quotes Loew to the effect that
Macropeza albitarsis, Mg., preys upon other small insects.

This list could no doubt be greatly extended by careful
observation, and it seems probable that all the members of the bare-
winged genera of Ceratopogoninae are normally predaceous in the
female sex. Evidently the various modifications of the legs, such
as swollen and often spiny femora, enlarged claws and spines on the
last tarsal segments, which the females of most of these genera
exhibit, are to be regarded as adaptations for holding their insect
prey. It is probable that these predaceous habits are primitive in
this sub-family, and that they have directly or indirectly led to the
more specialised blood-sucking habits of certain species and genera.

Although it 1s beyond the scope of this paper, attention may be
called in passing to the observations of Ingram, who found in West

* Noted in North Cornwall, June, 1922. This is the only fresh record I have to add to
the list previously published. .

+t Goetghebuer’s review (1922) of the Ceratopogoninae in Meigen’s collection has made
it clear that C. communis, Mg., the type of the genus, belongs to Kieffer’s genus Psilobelea ;
this name, therefore, falls as a synonym of Ceratopogon. As I have stated in a recent paper
(1921) I do not consider the differences between Ceratopogon (Psilobelea) and Isohelea are of
more than subgeneric value, hence I include J. lacteipennis and its allies also in Ceratopogon.

Africa the larvae of Forcipomyta ingrami, Carter (1919), attacking
mosquito larvae. This is, I believe, the only known instance of
predacity in a Ceratopogonine larva.

(2) Cases of blood-sucking, where the attacking midge sucks the
juices of its victim, without as a rule killing it, the victim in such
cases being generally much larger than the attacking species. It is
this class of phenomena with which I wish to deal more particularly
in the present paper. Following the example of Peyerimhoff (1917),
we may consider these midges in several groups, according to the
type of host which they attack.

I. SPECIES ATTACKING MOSQUITOES

A considerable number of observations have been made on the
relations between adult mosquitoes (generally Azopheles) and a
species of Culicoides which is widely spread in the Oriental region.
In a recent paper (1922) I have summarised these observations, and
have described the midge concerned as Culicoides anophelis. It
appears that the object of the Culicoides is to obtain engorged blood
from the abdomen of its host, though it has in some cases been found
to have attacked mosquitoes which were not engorged. At present
only this single species of Culzcoides 1s known to have these very
remarkable habits.

This extremely interesting case may be regarded in one of two
ways. It may be a development directly from a primitive predacity ;
the species having passed from a diet of (say) Chironomidae to one
of mosquitoes, and thence to the mammalian blood contained in the
body of its host. In this case it 1s easy to imagine that the midge
might follow its mosquito host to its feeding ground, and eventually
take to sucking blood itseif directly from the mammal, thus giving
rise to the blood-sucking habits now so general in the genus
Culicoides. The possibility of this having been the course of
development 1s somewhat strengthened by the fact that C. axophelis
appears to show some somewhat primitive characters, such as the
simpie wing-pattern and rather large radial cells. On the other
hand, it may be that the habit of obtaining blood from mosquitoes
is purely secondary, and derives from an ordinary direct method of
blood-sucking; this is, perhaps, most probable, since Lamborn’s
observations seemed to show that a blood meal was essential to the
production of a complete fertile batch of eggs.

22
II. SPECIES ATTACKING ADULT LEPIDOPTERA

One instance has been recorded (by Kryger, 1914, quoted also
by Knab, 1914) of a midge attacking a moth. The host was
Cidaria didymata, ..; the midge was not precisely identified, but
was stated by Knab to be apparently an undescribed species,
‘belonging in the neighbourhood of Ceratopogon murinus, Winn.’
In the hope of obtaining some further information concerning this
species, I wrote to Mr. Kryger in Denmark, and also to Messrs.
Aldrich and Béving in Washington, but only to discover that the
material had been lost.

A second very similar case was discovered by Professor Newstead
in North Wales in 1914, and I am greatly indebted to him for kindly
allowing me to examine and describe the material of this most
interesting find. While collecting at night with the aid of an
acetylene lamp, Professor Newstead came across a cabbage-white
butterfly whose wings were being attacked by nine specimens of a
Ceratopogonine midge. The butterfly was considerably damaged,
and as is shown by the accompanying photograph (fig. 1) the

Fic. 1. Pieris napt (slightly enlarged), victim of Forcipomyia (Euforcipomyta) paptlionivora,
Edwards. The left forewing, just below the costa and along both sides of the large vein, shows
the nature of the damage, caused by the midges

damage would seem to have been caused, at least in part, by the
attacks of the midges.* The latter appeared to be eating the wings

* Blood was seen exuding from the ruptured veins when the insect was captured; and
the scales on either side of the veins are stained russet-brown, due apparently to the exudation.
When first imprisoned the midges left their victim and swarmed over the glass lid of the
collecting box; but on placing them in the dark, they were found, two hours later, to have
resumed their attacks on the butterfly. (R. Newstead).

of the butterfly, though, as in the case of the Danish insects, they
may in reality have been sucking juices from the wing-veins,
especially if the blood was exuding from the broken ends of the
veins.

After a careful examination of the literature, I have come to the
conclusion that the midges collected by Professor Newstead belong
to an undescribed species, and I, therefore, name and describe it as
follows :-—

Lorcipomyia (Luforcipomyta) papilionivora, sp.n.

Head rather densely clothed with golden pubescence. [yes
practically touching, perfectly bare. Axtennae uniformly dark,
flagellum clothed with longish dark hair, nearly twice as long as
the diameter of the segments. First eight flagellar segments
together much shorter than the last five together (proportions 3 : 5).
First flagellar segment nearly globular, the next five slightly
transverse, seven and eight again practically globular, nine to
thirteen each nearly three times as long as broad, thirteen with a
nearly globular, nipple-like tip; one to eight each with rather long
and stout sense-bristles, difficult to observe. Palgz dark, the second
segment oval, broadest in the middle, not quite twice as long as
its greatest breadth, apical part not suddenly narrowed; last two
segments together as long as the second, the fourth a little longer
than the third. Second segment with a globular internal cavity
opening by a small round pore on the inner face. M@andzbles broad,
about three to five times as long as their greatest breadth, tip rather
bluntly rounded, with about twelve to fifteen small, equal-sized teeth
on one side; in the middle is an oval clear spot enclosing an
elongate dark mark, resembling that figured by Carter, Ingram and
Macfie in the genus Prionognathus. Mazxillae almost as long as the
mandibles, with about twenty-five fine regular crenulations, scarcely
teeth, on one margin. Hypopharynx rather elongate, oval, a little
over twice as long as broad, tip smooth. JZorax with the integu-
ment dull blackish, the humeral angles and the whole scutellum dull
yellow. Mesonotum densely covered with short, bright golden
pubescence mixed with longer, but not very long, brownish hair.
Scutellum similariy but less densely clothed, postnotum shining
black. Addomen rather narrow for the genus, dull dark brown,
uniformiy clothed with short blackish hair, S/ermathecae large,

nearly globular, necks practically without chitinisation. Cerct
dark. Legs slender, practically uniform in colour, rather dark
brownish, very hairy, the tibiae with some long hairs which are
about six times as long as the tibial diameter. On all the legs the
first tarsal segment is 2°5 times as long as the second. Empodia
well developed, almost as long as the claws. Wzxgs clothed rather
densely (but somewhat less densely than in most species of the
genus) with close-lying dark hair; most of the hair on the thick
veins golden, but mixed with some dark. Venation normal for the
genus: As in contact with #7, so that the first radial cell is
obliterated; second radial cell about twice as long as_ broad,
trapezoidal; petiole of median fork a little shorter than the very
oblique r-m; cubitus forking below end of costa, which reaches just
beyond the middle of the wing. /Halteres with the stem dark, the
knob white.

Length of body, 1°8 mm.; wing, 1°4 mm.

NORTH WALES: Ty Gwyn Farm, Aberhosan, Machynlleth,
found at 10.15 p.m. feeding on the wings of Pieris nafpi
(R. Newstead). Three 9 co-types in the British Museum, presented
by the collector ; six others in the collection of the Liverpool School
of Tropical Medicine.

This insect seems to have no very close ally among the European
species. By Kieffer’s table it will run down to F. formicaria
(Kieff.), which differs in having the first tarsal segments much
shorter, as well as in the palpal structure and other details. Other
European species which show some points of resemblance are
I’, hirta (Lundst.) and F. murina (Winn.), but none show the
combination of antennal and tarsal characters possessed by this
species. In both these respects the new species resembles Lasiohelea
velox (Winn.), but it has not the venation of the genus Lasiohelea;
it belongs to Malloch’s group Euforcipomyia, and bears a close
resemblance to the North American £. fusicornis (Coquillett) (see
below).

From the above remarks it will be seen that the specimens
collected by Professor Newstead might have been referred to as
‘an undescribed species belonging in the neighbourhood of
Ceratopogon murinus, ‘Ninn.,’ and it, therefore, seems not
improbable that the Danish specimens found by Mr. Kryger may
have belonged to the same species. In any case, it is interesting

to note that the only two records we have of midges attacking adult
Lepidoptera both refer to an insect belonging to the same group of
the genus //orcipomyia.

III. A SPECIES ATTACKING A SIALID

Malloch (1915) states that he has seen a specimen of Lw/orci-
pomyia fusicornis (Coquillett) which was taken attacking a Sualid
(Chauliodes sp.). In view of the large size of the victim, this must
be classed as a case of blood-sucking rather than of predacity. It
is not stated what part of the Sialid was attacked, but it is evident
that we are here dealing with a very similar case to the two last
considered, Chauliodes being a large-winged, rather soft-bodied
insect, comparable with a moth. It is, therefore, of special interest
to note that £. fusicornis, according to Malloch’s description,
bears a very close resemblance to the species just described as
I’. papilionivora. In fact, it 1s not impossible that the two may be
conspecific, though it seems unsafe to identify a European with a
North American form without actual comparison of material.

Although I do not consider Euforcipomyia to be generically
distinct, it may be retained as a sub-genus in the sense in which
Malloch proposed it: 1.e., to include the species of Forcipomyia
which have the first hind tarsal segment markedly longer than the
second, reserving Forcipomyia (s. str.) for those species in which the
first is shorter, or at most slightly longer, than the second. This is
not the sense in which Kieffer has used the name, but seerns to be
the correct one, since the type species of orczpomyia is bipunctatia,
L. (¢richoptera, Mg.), not albipennis, Mg., as stated by Kieffer.

IV. SPECIES ATTACKING CATERPILLARS

A number of cases of midges attacking caterpillars have been
recorded from time to time. Most of these were referred to by Knab
(1914), the cases he mentioned being as follows :—

SPECIES Host OBSERVER
Forcipomyia Melanchrota
propinqua (Will.) eeometroides (Waker) Baker (Cuba)
(Geometridae)
F. squamosa, Lutz.* Sphingid (undetermined) ‘Townsend (Peru)
F. sp. Sphingid (undetermined) Barbiellini (Brazil)
I. crudelis, Knab.t Not stated Urich (Mexico)

F. erucicida, Knab. Erinnyis ello L. (Sphingidae) Mosier (Florida)

* Specific name given by Lutz (1914).

t This specific name is preoccupied by F. crudelis (Karsch), but I refrain from proposing
a substitute because the descriptions appear to indicate that the species is almost certainly
identical with F. tropica, described by Kieffer (1917) from Costa Rica.

All the above-named species of /’orcipomtyia belong to that group
of the genus in which the female tibiae are devoid of scales, and the
second segment of the hind tarsi is at least twice as long as the
first. The same remark 1s true of three other species which were
not known to Knab, and are discussed below. It would seem,
therefore, that the habit of attacking caterpillars is a very special
one, restricted to this group of the genus Forczpomzyia.

lorcipomyia crudelis (Karsch, 1886)

This species was described from a single female found by Karsch
sucking a saw-fly larva in the neighbourhood of Berlin. He remarks
that his attention was called to the larva by the movements which it
made in endeavouring to dislodge its tormentor, and that the latter
had its mouth-parts so firmly fixed in the body of its victim that it
did not loose its hold even when the pair were placed in the cyanide
bottle. /. crudelzs has not been recognised since Karsch described
it, but it is evidently very closely related to /. pallida (Winn.),
I, brevimanus (Lundst.) and F. alboclavata (Kieffer).

Forcipemyia hirtipes (de Meij.)

T'wo females of this species were found by Mr. J. C. F. Fryer
(recorded by me, 1913), at Peradeniya, Ceylon, each sucking a larva
of Papilio clyttia. F. hirtipes, it is interesting to note, closely
resembles the European F. alboclavata, showing only very slight
differences in the proportions of the palpal and tarsal segments.
The Ceylon specimens do not agree with de Meijere’s description,
as regards the middle tarsi; but, as he has informed me, the
description is incorrect. In reality, he says ‘the mid-tarsal
segments have about the same proportions as the hind, viz., in ¢
about 8s Zo fe1giratie Beane labout o% 25 mi. OF: 73/neHefalso
informs me that though the antennae of the type 2 are mutilated,
in another specimen the proportion of the first eight to the last five
flagellar segments is about 27 : 38. These proportions are about the
same as in the Ceylon specimens.

Forcipomyia alboclavata (Kieffer, 1919), (canaliculata,
Goetghebuer, 1920)

The British Museum possesses three 9? 9 of this species from
Taterafuered, Hungary, 1906 (Hon. N. C. Rothschild), on which

the donor sent the following note :—‘ Sitting on the backs of larvae
of Deilephila galii (which were extremely common in a large field
near Taterafuered), and appearing to eat some secretion from their
skins.” The specimens have the second hind tarsal segment 2°5
instead of only twice as long as the first, but otherwise agree with
Keiffer’s description, and I have no hesitation in quoting the
synonymy as above.

Ff’, alboclavata has been found in Scotland (Arran) as well as
Belgium, but its habits in these countries have not been observed.

V. SPECIES ATTACKING OIL-BEETLES

Peyerimhoff (1917) has given an interesting account of the
relations between a Ceratopogonine midge (at present undetermined)
and the oil-beetle Aleloe majalis, L., m Algeria. The flies, he says,
pursue these large beetles in little swarms, and without incon-
veniencing them in any. way, feed upon their yellow blood.
M. de Peyerimhoff informs me that the flies are now in the hands
of Professor J. J. Kieffer, who believes that they represent a new
species. *

More recently, a second similar instance, this time from
Denmark, has been recorded by Hansen (1921). I am indebted to
my friend, Mr. J. P. Kryger, for the following translation of
Mr. Hansen’s note :—

‘A gnat attacking a Meloe—2oth May, 1921. I saw a Meloe
proscarabaeus crawling along a walk in the wood of Ulvlyst
(Denmark). A little swarm of gnats hovered over the beetle and
sometimes attacked it, especially on the soft skin between the first
and second thoracic segments. The beetle was seriously affected by
the gnats biting, and rubbed its sides with its hind legs, but
without getting rid of its tormentors. When I put the collecting
bottle over the beetle two gnats were sitting on its back, but as it
tumbled in twelve gnats appeared in the bottle. The remaining ten
must have been sitting on the underside of the beetle.’

Mr. Kryger has further been so good as to obtain for me the
loan of the specimens captured by Mr. Hansen, which had been

* Since this was written I have received, through the kindness of Professor M. Bezzi, a
number of the specimens originally collected by M. de Peyerimhoff. Without trespassing on
ground to be covered by Professor Kieffer, I may remark that these specimens represent a
species which is extremely nearly related to Atrichopogon rostratus (Winn.), a fact which is of
much interest in view of my determination of the Danish specimens.

presented by the collector to the Zoological Museum at Copenhagen.
Upon examination of the flies I find that they belong to the species
Atrichopogon rostratus (Winn.), all, of course, being females. The
purpose of the formidable proboscis possessed by this species thus
becomes apparent for the first time, for neither it nor any other
member of its genus has been known either to bite warm-blooded
animals or to prey upon other small insects. But, as in the case of
many other midges with strong food preferences, the diet of
A. rostratus is not confined to the blood of Meloe, but consists
partly of vegetable substance (honey, or perhaps pollen). All the
adult specimens of this midge which I have found myself have been
taken on the flowers of umbellifers (Axzgelica and Heracleum),
often in company with great numbers of some other species of
Atrichopogon.

VI. A SPECIES ATTACKING A PHASMID

Williston (1908) mentions a minute fly which was found in the
West Indies ‘closely applied to and apparently sucking the juices
from the antennae of a Phasmid.’ He considered the specimen to
represent a new genus of Szzulzidae, but the figures which he gives
indicate rather a Ceratopogonine midge. The available evidence is
insufficient to place this species generically, though if Williston’s
figure of the wing is accurate, 1t would not seem to fit very well
into any known genus. The specimen is not among the West
Indian collections in the British Museum which were studied by
Williston, and I am informed that it cannot be traced in those parts
of his collection which are now 1n Washington and New York.

REFERENCES

Baxer, C. F. (1907). Remarkable habits of an important predaceous fly (Ceratepogon eriophorus
Will). U.S. Dept. Agr., Bur. Ent. Bull. 67, pp. 117-118.

Carter, H. F. (1919). New West African Ceratopogoninae. Ann. Trop. Med. & Parasit.,
Vol, XU puizo7.

Epwarps, F. W. (1913). New and little-known Diptera Nematocera from Ceylon. Ann. Mag.
Nat. Hist., Vol. XII, (8), p. 201.

——— (1920). Some records of predaceous Ceratopogoninae (Diptera). Ent. Mo. Mag.,
Vol. VI (3), pp. 203-205.

(1921). Diptera Nematocera from Arran and Loch Etive. Scottish Nat., p. 124.

(1922). On some Malayan and other species of Culicoides, with a note on the genus
Lasiohelea. Bull. Ent. Res., Vol. XIII, pp. 161-163.

Gorrcuesuer, M. (1921). Ceratopogoninae de Belgique. Mem. Mus. d’Hist. Nat. Belg.,
Vol. VII, p. 110.

(1922). Etude critique des Ceratopogon de la Collection Meigen. Bull. Soc. Ent.
Belg., Vol. IV, p. 52.

Hansen, J. (1921). En Myg, der angriber en Oliebille. Flora og Fauna, p. 106.

Karscu, F. (1886). Ein neues markisches Dipteron. Berl. ent Zeit., Vol. XXX, Sitzber,
pp. XVII-XVIII.

Kirrrer, J. J. (1917). Chironomides d’Amérique. Ann. Mus. Nat. Hung., Vol. XV, p. 297-
(1919). Chironomides d’Europe. Ann. Mus. Nat. Hung., Vol. XVII, p. 12.
(1922). Observations biologiques sur les Chironomides piqueurs. Arch. Inst. Pasteur.
Afr. Nord., Vol. I, p. 387.

Knap, F. (1914). Ceratopogoninae sucking the blood of caterpillars. Proc. Ent. Soc. Washington,
Vol. XVI, pp. 63-66.

Krycer, J. P. (1914). En Myg, der angriber en Sommerfugl. Ent. Medd., pp. 83-88.
Lutz, A. (1914). Mem. Inst. Oswaldo Cruz, Vol. VI, p. 88.

Mattocn, J. R. (1915). Some additional records of Chironomidae for Mlinois. Bull. Jil.
State Lab. Nat. Hist., Vol. XI, p. 315.

PryeriMuHorr, P. pe (1917). Ceratopogon (Dipt. Chironomidae) et Meloe (Col. Meloidae). Bull.
Soc. Ent. France, pp. 250-253.

Witutston, S. W. (1908). Manual of North American Diptera. 3rd ed., p. 147.

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ON SOME STRONGYLID LARVAE IN
THE HORSE, ESPECIALLY “THOSE -OF
CYLICOSTOMUM

BY
J. E. W. IHLE, D.Sc.
AND

eee Na COIL) ds LPC:
(Zoological Laboratory of the Veterinary College, Utrecht, Holland)
(Received for publication 5 December, 1922)

The adult stage of species of Cylicostomum found in the large
intestine of the horse has been extensively studied by Looss and,
more recently, by Boulenger, and Yorke and Macfie (cf. [hle, 1922),
but very httle is known as yet of the development of these species in
the body of the horse.

The larvae of Cylicostonium are to be found in large numbers
in. the mucosa of the caecum and colon of the horse. They were
first found by Dick (1836) and described by Knox (1836). By
Diesing (1851, Vol. II, p. 332) they were mentioned under doubtful
species as Nematoideum equi caballi. T. Spencer Cobbold
(1874, p. 85) describes and figures Cylicostomum larvae as adult
Nematodes under the name T77ichonema arcuata, but the next year
(1875, p. 241) he states that 77zchonema is only the larval form of
Cylicostomum (* Strongylus tetracanthus’).

Short descriptions and sketches of the larvae, encysted in the
mucosa of the large intestine, are also to be found in Leuckart
(1876, p. 445), Cobbold (1886, p. 288) and Giles (1892, p. 15,
Pl. Ill, figs. 16, 18). The last-mentioned author thought he had
found a free-living Rhabditis generation of Cylicostomum. ‘This
mistake, which has been made repeatedly and also recently, when
the development of different Nematodes was traced, is due to the
fact that the cultures of larvae were infected with free-living
Nematodes.

An investigation of Cuillé, Marotel and Roquet (1913), dealing
with our subject, is of more importance than the older publications
above mentioned. These authors distinguished three types of larvae,
living in the mucosa of the large intestine of the horse and considered
as belonging to Cylicostomum:—({1) ‘La larve oesophagostomi-
forme,’ with mouth-capsule and dorsal tooth and having a length
of 2 to 5 mm.; (2) ‘la larve metastrongyliforme,’ without mouth-
capsule (length 800% to 2 mm.); and (3) ‘embryons,’ without
recognisable internal structure (length 3004 to 800%). They showed
that the ‘larve oesophagostomiforme’ passes over into the juvenile
Cylicostomum by a moult.

Recently a part of the development of Cylicostomum insigne was
shortly described by Boulenger (1921), who figures small larvae
(6 to 7 mm. in length) and Jarge larvae (up to 11 mm. in length),
both agreeing with the ‘larve oesophagostomiforme’” of the French
authors. In the larger larvae the adult mouth-capsule makes its
appearance, which represents the preparation for the last ecdysis.

We ourselves have examined a large number of larvae,
partly collected by the Commission appointed to inquire into
Sclerostomiasis in Holland, and partly by ourselves in the horses
dissected in the Anatomical Institute of the Veterinary College at
Utrecht. All the larvae were found in the mucosa of the large
intestine, though a small number were met with free in the lumen
of the intestine.

The larvae examined by us can be divided into different types,
to be described in subsequent pages. They belong for the greater
part to Cylzcostomum, a few perhaps to 77iodontophorus; others
were not identified. In addition we found a few very small larvae
without recognisable internal structure. They agree with the
so-called ‘embryons’ of Cuillé, Marotel and Roquet (1913, p. 8 of
reprint), and were only obtained by us in a few cases by scratching
the mucosa of the large intestine. We have not yet studied these
forms in detail, but we do not think that these small worms
(according to the French authors measuring 300m to 800, in length)
must be considered to belong to the genus Cylicostomum, because
it follows from the investigations of A. Albrecht (1909) and of
De Bheck and Baudet (not yet published) that the larvae of
Cylicostomum infecting the horse are much more differentiated.

33
CYLICOSTOMUM LARVAE

All Cylicostomum larvae, found by us in the mucosa or in the
lumen of the intestine, show a cup-shaped larval mouth-capsule,
which has been already described by Cobbold (1874, p. 86). In
agreement with this author (1886, p. 288) we will call this larval stage
Trichonema stage. The name ‘larve oesophagostomiforme’ must
be rejected, as these larvae do not in any particular agree with
Oesophagostomum.,

The cuticle is ringed. The cuticle surrounding the circular
mouth-opening may also be called mouth-collar here; this larval
mouth-collar, however, 1s much less developed than the adult one.
The mouth-opening is generally surrounded by six papillae.
External and internal leaf-crown are absent. The mouth-capsule
is either sharply marked off from the mouth-collar or passes
gradually over into it. In the middle the mouth-capsule is mostly
wider, and possesses a thicker wall than posteriorly and anteriorly.
Especially near the mouth-opening, the wall is very thin. The
anterior part of the mouth-capsule is mostly provided exteriorly
with a collar, often strongly developed, and which we will call
mouth-capsule collar. It is divided into six lobes, which have a
crescent shape and are almost perpendicular to the outer surface of
the mouth-capsule. Between every two lobes a head-papilla is to
be found.

An oesophageal funnel, in which the three sectors of the
oesophagus continue, is present. The dorsal sector 1s always
provided with a tooth, more or less protruding into the lumen of
the mouth-capsule. The cuticular lining of the anterior margin of
the oesophageal funnel shows a circular thickening, adjacent to the
mouth-capsule. We will call this thickening the funnel-ring; it is
directed to the exterior.

The oesophagus is cylindrical in shape and somewhat swollen
posteriorly. Where the oesophagus passes over into the mesenteron
three valves protrude into the lumen of the intestine. A nerve-ring,
surrounding about the middle of the oesophagus, is present.

The mesenteron is composed for the greater part of a dorsal and
a ventral row of alternating, polynuclear cells, which are mostly
pigmented. In the anterior part of the mesenteron the cells are
always much flatter than in the posterior part. In the anterior part

the cell-limits run transversely or directed obliquely to the front;
so that the lateral parts of the cell-limits are situated more anteriorly
than the dorsal and ventral parts.

The very short rectum opens into the exterior through the anus,
situated at a small distance from the sharp posterior extremity of
the body. :

Sometimes, but not always, the larvae living in the mucosa are
red in colour. It appears that in the few cases examined by us
the whole body, the pigmented intestine excepted, is red. When
such a larva is pricked, a red fluid is emitted. The juvenile
specimens, living in the lumen of the colon and caecum, may also
show this colour, but, as in the case of the larva, the intestine was
not red in the specimens examined by us. Prof. B. Sjollema and
Miss J. E. van der Zande were so kind as to analyse microchemically
and spectroscopically a few juvenile specimens of Cylicostomum |
wnsigne for us. ‘The fluid appeared to be due to oxyhaemoglobin.

We assume that the larvae living in the mucosa feed on blood
at least during a part of their life. Boulenger (1921, p. 324) found
these larvae in cysts, filled with blood; this is not always the case,
however. As mentioned above, the red colour was not observed in
the cells of the intestine of the larvae. We suppose that the larvae
had fed on blood in an earlier period; consequently the red colour
must have disappeared already from the intestinal wall, but not yet
from the rest of the body. Further, we are of opinion that the red
colour of the adult worm is the consequence of the larvae having
fed on blood, for the adult Cylicostoutum feeds on the contents of
the large intestine of the host and not on blood. After dissection
these worms are never found attached to the mucosa of the host’s
intestine.

The larvae, which we consider to belong to the genus
Cylicostomum, can be divided into two types, to be described
below. Not much importance must be attached to the dimensions
indicated, as larvae of numerous species are brought together which
when adult differ strongly in size. We cannot state to which species
of Cylicostomum these different types belong, because we have not
at our disposal a large enough number of moulting specimens.

35
Cylicostomum Larva. Type A (fig. 1).

To this type the smallest larvae of the genus Cylicostomum are
considered to belong, having a length of 3 to 475 mm. and a
maximum thickness of 110“ to 200#. The mouth-margin is smooth.
Around the mouth-opening the cuticle is thick. Head-papillae
could not be observed. A mouth-capsule collar was not found by
us. The length of the mouth-capsule, including the mouth-collar,
varies from 204 to 28pn.

Fic. 1. Anterior extremity of a Cylicostomum-larva, type A, seen from right side. x 540
(x }#). d.t.—Dorsal tooth; 0es.—Oesophagus; c.—Cuticle; /.m.c.—Wall of the larval
mouth-capsule; f.r.—funnel-rings; /.v.s.—Latero-ventral sector of the oesophageal funnel.

In this type the oesophageal funnel also possesses three sectors.
The dorsal sector always possesses a tooth, varying in size; the two
latero-ventral sectors are rounded or bear an inconspicuous tooth,
which never protrudes as far into the lumen of the oral capsule as
the dorsal, large tooth. The length of the oesophagus varies from
250@ to 350". The distance from the anus to the posterior
extremity of the body is 80 to 130u.

This type 1s very common.

Cylicostomum Larva. Type B (fig. 2).

Another type (B) is also of frequent occurrence. It differs from
Type A in being of a larger size and in possessing a mouth-capsule
collar. The length is 7°5 to 12°5 mm., the maximum thickness

420n to 580. Six head-papillae are present, agreeing as to
arrangement with those of the adult specimens; so there are two
lateral and four sub-median papillae. The oral margin is mostly
somewhat incised near the six papillae. Length of the mouth-
capsule, including the mouth-collar 55 to 65m. At one-third of
the length of the mouth-capsule from the posterior margin the wall
of the oral capsule is thickest. The wall of the mouth-capsule
becomes thinner anteriorly and passes gradually over into the mouth-
collar. The mouth-capsule collar is very well developed and
situated immediately under the cuticle of the anterior part of the
body. Here the cuticle 1s somewhat thickened.

Fic. 2. Cylicostomum-larva, type B, seen from right side. x 435 (x 2). d.t.—Dorsal
tooth; m.c.c.—Mouth-capsule collar; s.—Septum; /.7.—Funnel-ring.

The oesophageal funnel is conspicuous and bears a cuticular
lining of variable thickness. The funnel-ring is well developed.
In some cases the border between mouth-capsule and funnel-ring is
irregular or undulating. Sometimes the funnel-ring possesses a
circular groove at its outer surface, so that in optical section it
appears as a double ring. In this type, too, the dorsal sector of
the oesophagus is continued as a tooth, protruding into the lumen
of the oral capsule; this tooth is relatively not so large as in Type A.

The latero-ventral sectors are truncated anteriorly, or become lower
and lower, to end at the funnel-ring. At the bottom of the grooves
by which the sectors are separated the cuticle is thickened, just as in
the adult worm. The oesophagus measures 550 to 650, in length;
the distance from the anus to the extremity of the body is 190m
to 220.

We consider that the larvae belonging to Type B represent a
more developed stage of Type A, because we have found several
larvae with the rudiments of the mouth-capsule collar (fig. 3); these

Fic. 3. Cylicostomum-larva, intermediate between types A and B, seen from dorsal side.

x 540 (x #). m.c.c—Mouth-capsule collar; d.t.—Dorsal tooth; fr.—Funnel-ring ;
l.o.—Lining of the oesophageal funnel.

larvae are of a size intermediate between those of Types A and B.
Length 5 mm. to 6°5 mm., maximum thickness 350u; length of the
mouth-capsule, including the mouth-collar, 424; oesophagus 540
long.

The last ecdysis (figs. 4, 5).

The Zvichonema stage passes over into the juvenile worm, living
in the lumen of the large intestine, by a moult. In agreement with
the development of other Nematodes, we assume that this moult is
the fourth and last. The ecdysis itself takes place in the intestinal
lumen. |

The moult begins with the formation of a cavity around the larval
mouth-capsule (fig. 2). We consider that one continuous cavity is

present from the beginning. Boulenger (1921, p. 325) mentions a
series of cavities. However, according to Looss (1897, p. 925), two

cavities (a dorsal and a ventral one) are formed in the larva of the

fourth stage of Axcylostoma. Later on these cavities unite to form

a circular lumen.

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Fic. 4. Cylicostomum-larva, type B, seen from left side, with rudiment of the cavity of
the adult mouth-capsule. x 290 (x #). s.—Septum; a.m.c.—Wall of the adult mouth-
capsule; m.c.c.—Mouth-capsule collar; ¢.a.m.c.—Cavity of the adult mouth-capsule.

When the cavity makes its appearance, it is narrow at the front
and a little wider backwards. Later on the anterior part of this

cavity extends to the mouth-capsule collar. In the posterior part,

which extends almost to the oesophagus, we see a granular substance,

which seems to form a thin layer (fig. 2, s.) about at the level of the

posterior margin of the mouth-capsule. This layer corresponds with

the definitive anterior side of the mouth-capsule of the adult worm.
This septum (fig. 4, s.) gradually becomes thicker, possibly formed
by the granular substance mentioned above, while the cavity lying
behind this septum, and in the beginning filled up with this
substance, becomes empty and extends simultaneously backwards.
This cavity, situated behind the septum, is the lumen of the adult
mouth-capsule. At the periphery of the septum the definitive

mouth-collar and the definitive head-papillae develop (fig. 5). The
cavity mentioned gradually widens and peripherally begins to
form the wall of the adult oral capsule. Now this circular cavity
surrounds the anterior part of the oesophagus (fig. 4).

Fie. 5. Cylicostomum-larva, type B, moulting. x 290 (x #). — m.c.—Mouth-collar;
o.f.—Larval oesophageal funnel; m.c.c.—Mouth-capsule collar; 1.m.c.—Wall of the
larval mouth-capsule; (f.7.—funnel-ring; 4.m.c.—Wall of the adult mouth-capsule ;
oes.—oesophagus.

Meanwhile the cuticle of the adult worm is formed below the
provisional one. Before the ecdysis proper the oesophagus loosens
itself from the cuticular lining of its funnel (fig. 5). In earlier stages
the oesophagus tapers to the anterior extremity and ends at the
funnel-ring (fig. 4). But at this stage it becomes truncated in front.
Now the lumen of the mouth-capsule is situated before the
oesophagus, whereas the anterior part of the latter was formerly
surrounded by the adult mouth-capsule. Simultaneously the mouth-
capsule and the funnel-ring, which remains connected with the
cuticular lining of the provisional oesophageal funnel, separate.
The posterior margin of the mouth-capsule and the anterior margin

of the funnel-ring remain connected by a thin membrane, of which
the origin is difficult to trace. Boulenger (1921, fig. 5 4) also figures
it, without describing it. In the moulting specimen sketched by
Cuillé, Marotel and Roquet (1913, fig. 17, 7), in which the adult
mouth-capsule and mouth-collar have developed completely, the
funnel-ring and the provisional mouth-capsule are still connected.
In the specimen sketched by us the external leaf-crown is already
visible, but not indicated in the figure.

OTHER LARVAE

Besides the larvae Types A and B (Cylicostomum), we also
found some other types, which we were unable to identify. Short
descriptions, dealing only with the differences between these types
and the larvae of Cylicostomum, will now be given.

LaRvA. Type C (fig. 6).

This type and the following Type D do not differ essentially
from the larvae of Cylicostomum. ‘Type C was found only once in
the mucosa of the large intestine; but three specimens could be
investigated. The length is 4°5 to 6.6 mm., the maximum thickness

| reieetere?.
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Fic. 6. Larva, type C, seen from right side. x 310 (x 3). 1.b.p.—Lateral head-

papillae; d.t.—Dorsal tooth; m.c.c.—Mouth-capsule collar; 1.m.c.—Wall of the larval
mouth-capsule; f.7.—Funnel-ring.

225m to 380”. The mouth-opening is circular, its margin 1s
delicately denticulated. A mouth-collar is present, of which the side
directed to the body-axis possesses a layer passing over into the
anterior margin of the mouth-capsule. (In the figure the limit
between mouth-collar and mouth-capsule is not indicated.) The
head-papillae are present. The oral capsule is short and very wide;
it is 52m to 65m in length, the mouth-collar included. The mouth-
capsule collar is well developed and implanted in the anterior half
of the mouth-capsule.

The oesophageal funnel 1s wide and bears a dorsal tooth. The
latero-ventral sectors are smooth. The funnel-ring is very long at
the dorsal side, shorter, however, than at the ventral side, where its
length is equal to that of the mouth-capsule. In optical section this
ventral part especially has the shape of an inverted Y, which encloses
-a part of the musculature of the oesophagus. The latter is short
and thick, 435m to 550m in length, consequently measuring one-tenth
to one-twelfth of the total body-length.

The posterior extremity 1s rounded. In one of the specimens
examined, the part of the body situated behind the anus becomes
gradually thinner; the distance from the anus to the posterior
extremity is considerable (380) here. In two other specimens the
thickness of this part diminishes suddenly at some distance behind
the anus; consequently the body ends in an almost cylindrical point.
In these cases the distance from the anus to the posterior extremity
is 155¢ to 1804. Possibly these are sexual differences.

LarvA, Type D (fig. 7),

We found this type five times (only a few specimens) in the
mucosa of the large intestine. We do not know whether this type
and also the former (Type C) belong to Cylicostomum. Length 3°5
to 5°I mm., maximum thickness 120 to 190¢. The head-papillae
are present. The cuticle is swollen around the mouth-opening ; here
the part of the cuticle directed to the body-axis possesses a particular
layer, which passes over into the mouth-capsule. The latter is
25m to 27H long, the cuticle surrounding the mouth-opening
included. Posteriorly its wall increases in thickness. A slightly
developed mouth-capsule collar is present, lying immediately against
the cuticle.

The dorsal sector of the oesophageal funnel bears a large tooth,
protruding far into the lumen of the mouth-capsule. At the dorsal
side its anterior margin possesses a small point and at the ventral
side a large one. Each of the latero-ventral sectors bears a small
tooth with one point. Moreover, the well-developed funnel-ring

Fic. 7. Larva, type D, seen from right side. x 540 (x #). d.t.—Dorsal tooth ;
v.t.—Ventral tooth; /.m.c.—Wall of the larval mouth-capsule; f.7.—Funnel-ring.

possesses medio-ventrally a pointed tooth, being directed anteriorly
(fig. 7, v.¢.). Peripherally the funnel-ring does not protrude
markedly. ‘The oesophagus is long (400m to 435m), being one-ninth
to one-twelfth part of the body-length. The mesenteron agrees with
that of Cylzcostomum larvae. The anus is situated 105m to 115¢u
from the posterior extremity of the body.

DAR WAgS ey pes Pea tro ae

We found this type only once in four specimens in the lumen of
the large intestine. We consider that. these larvae belong to
Triodontophorus. It is, however, very remarkable that we found
this type only once in our comprehensive material, as two
Triodontophorus species are common in Holland, and sometimes
inhabit one host in large quantities.

The length of these four larvae is 7°6 to 8°5 mm., the maximum
thickness 310% to 365“. The mouth-opening is circular and
surrounded by a thin mouth-collar, finely and _ longitudinally
striated, and resembling an extremely little developed external leaf-
crown. The six head-papillae are distinctly visible. The length of
the mouth-capsule (including the mouth-collar) is 654 to 82u. The
mouth-capsule is wide, cup- or barrel-shaped, and sharply marked

Fic. 8. Larva, type E, seen from right side. x 335. (x #). d.t.—Dorsal tooth;
f.r.—Funnel-ring; — m.c.c.—Mouth-capsule collar.

off from the mouth-collar. In some of the specimens it lies
immediately against the cuticle. (The space between cuticle and
mouth-capsule in the specimen figured is possibly due to the
preservation.) The mouth-capsule collar reaches very far posteriorly
beyond the equator of the mouth-capsule. Around the posterior
part of the latter a circular cavity is already visible: the rudiments
of the lumen of the adult mouth-capsule.

The oesophagus is long (750 to $20), being about one-tenth
of the body-length. The oesophageal funnel is well developed. It
bears three large, pointed teeth, which protrude far into the lumen
of the mouth-capsule. The dorsal tooth is a little larger than both
the latero-ventral teeth. A funnel-ring is present. The distances
from the anus to the rounded posterior extremity of the body is in

. 44

two specimens respectively 100” and 110m, in both other specimens
respectively 220 and 240; possibly these are sexual differences.
We suppose that these larvae belong to T7iodontophorus, because
the oesophageal funnel bears three large teeth protruding into the
lumen of the mouth-capsule, this characteristic being present among
the adult Strongylids of the horse, in 77zodontophorus only. More-
over, the great length of the oesophagus of this type agrees with the
long oesophagus in 7. intermedius and T. brevicauda. For the
rest, no other larva was present in our material which could be
considered to belong to 77zodontophorus on better grounds.

LaRVA. Type F (fig. 9).

Besides the larvae described above, which all possess a well-
developed mouth-capsule, we found in the mucosa of the large |
intestine of the horse in one case one larva without mouth-capsule.

Length 3°7 mm., maximum thickness 190m. In front of the
oesophagus, having a length of 350, is a tube-shaped mouth-cavity,
projecting slightly above the level of the anterior extremity of the
body and being spherically swollen posteriorly.

We cannot decide whether this larva is identical with the ‘larve
metastrongyliforme ’ of Cuillé, Marotel and Roquet, which, however,
is shorter (length 800% to 2 mm.) than the specimen found by us.

tL ot¢
=

TTT.

Fic. 9. Larva, type F. x 340 (x #). p.m.co.—Provisional Mouth-cavity ; 0¢s.—Oecesophagus.

Possibly both the larvae observed by the French authors and the
specimen found by us are identical with a larva encountered by
Leuckart (1876, p. 446) in the mucosa, being I mm. long and
differing ‘durch die Abwesenheit des Mundbechers, dessen Stelle
durch einen schlanken und diinnhautigen Chitincylinder vertreten
war, wie bei den ersten parasitischen Jugendzustanden des Dochmius
trigonocephalus. Die Umwandlung in die Form mit Mundbecher
geschieht durch eine Hautung, die schon bei Exemplaren von
1°5 mm. vollendet ist.’ If the supposition above made proves to be
correct, Type F represents the third larval stage of Cylicostomum,
though the differences between the encysted larvae (larvae of the
third stage, enclosed in the cuticle of the second stage) and Type F
are conspicuous.

REFERENCES
Avsrecut, A. (1909). Zur Kenntnis der Entwicklung der Sklerostomen beim Pferde.
Zeitschr. f. Veterindrkunde, Vol. XXI.
Boutencer, C. L. (1921). Strongylid parasites of horses in the Punjab. Parasitology, Vol. XIII.

CopsoLp, T. Spencer, (1874). Observations on rare parasites from the horse. Veterinarian,

Vol. XLVII.

——w— (1875). Epizoéty in the horse, more especially in relation to the ravages, produced
by the four-spined Strongyle (Szrongylus tetracanthus). Veterinarian, Vol. XLVIII.

(1886). Description of Strongylus arnfieldi (Cobb.), with observations on Strongylus
tetracanthus (Mehl.). ourn. Linn. Soc. (Zool.), Vol. XIX.

Curtt#, Marorer et Roguer (1913). Nouvelle et grave entérite vermineuse du cheval: la
cylicostomose larvaire. Bull. Mém. Soc. Sc. Vétérinaire, Lyon.

Dirstnc, C. M. (1851). Systema Helminthum, Vol. II.

Gitrs, G. M. J. (1892). Some observations on the life-history of Sclerostomum tetracanthum
Diesing and on Sclerostomiasis in equine animals, in connection with a so-called out-
break of ‘ Surra’ at Shillong. Scient. Mem. by Med. Officers of the Army of India. Pt. 7.

Inte, J. E. W. (1922). The adult Strongylids (Sclerostomes), inhabiting the large intestine
of the horse. Report of the Commission appointed to inquire into the Sclerostomiasis in
Holland. 1. Zoological part, Vol. I.

Knox (1836). Remarks on the lately discovered Entozoa infesting the muscles of the human
body ; with some observations on a similar animal found beneath the intestinal membrane

of the horse. Edinburgh Med. and Surg. ‘fourn., Vol. XLVI.

Lrucxart, R. (1876). Die menschlichen Parasiten und die von ihnen herriihrenden Krank-
heiten, Vol. II.

Looss, A. (1897). Notizen zur Helminthologie Egyptens. II. 3. Die Lebensgeschichte des
Anchylostomum duodenale (Dub.). Centralbl. Bakt. Parasitenk. 1 Abt., Vol. XXI.

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AVIAN CESTODES FROM NEW GUINEA

II. CESTODES FROM CASUARIFORMES

DR. ALEXANDER KOTLAN
From the Royal Hungarian Veterinary College, Budapest

(Received for publication 5 December, 1922)

As it has been pointed out in the first part of this paper (Kotlan,
1921), the worms described in both the former and present notes, as
. well as those which will be described subsequently, belong to a rather
large collection of parasites which were partly sent, partly brought
back, by the Hungarian naturalist, Lewis Bird, from the formerly
German New Guinea, during the years 1897-1899.

The intestinal parasites of birds belonging to the Casuariformes
are represented in this collection by numerous Cestodes from
Casuarius picticollis, Sclat. A hasty examination of these worms
showed that they all belong to the family of Davaineidae. On
account of external features two species could be distinguished, a
larger and a smaller one, both belonging to the genus Davainea,
R. BI. (s.1.). Until now only one representative of this genus was
known from Casuariformes,* viz., D. australis (Krabbe, 1869),
from Dromacus novae hollandiae. This species, however, is easily
separated from the two above apparently undescribed forms.

La AINA. (S) lo GAS. UA Riles spin:
Host: Casuartus picticollis, Sclat.
Locality: Erima and Sattelberg.

The majority of the worms collected from this host—about
two hundred more or less developed specimens—belong to this new

* Meggitt (1921) quotes in his key to the species of Davainea two ‘ Davainea, sp. nov., Vevers,

1920’ from Casuarius uniappendiculatus Blyth. I have been unable to obtain Vevers’ paper
(Proc. Zool. Soc., 1920.).

species. The strobilae, coming from the two above-mentioned
localities, exhibit in their external appearance a well marked
difference, for those from Sattelberg are much more contracted and
have a shorter, almost cylindrical body, while those from Erima are
more stretched and thus longer in size. The largest specimens
measure 34 cm., the greatest width (3 mm.) occurs in the posterior
part of the strobila. The worms, which are in an expanded
condition, bear a well marked scolex, which is short and approxi-
mately square, its diameter being 1 to 1°2 mm., while in contracted
worms it is not clearly marked off from the strobila. It also happens
sometimes that the anterior end of the strobila bears by means of
unequal contraction a pseudoscolex-like thickening of 3 to 5 mm.
length, with the true scolex at the end, as is shown in fig. 1. The

Fic. 1. Davatnea casuarit, n.sp. Showing the extremely contracted anterior end of
the body with the scolex. x 17.

rather muscular rostellum measures 0'5 mm. in breadth, and is armed
with two hundred and fifty very large hammer-shaped hooks, which
are arranged in two rows. The hooks of the anterior row are 48p
to 54m, those of the posterior row 40m to 46m in length. As far as
I am aware, there is only Houttuynia struthionis (Houtt.) which has
larger hooks, all the other members of the genus Davainea (s. 1.)
bearing smaller ones. In the following table are enumerated some

Davainea species the rostellar-hooks of which are comparatively the
largest ones and measure over 20 in length :—

No. Length

of rostellar-hooks

Species Host

Houttuynia struthionis* (Houtt.) ... —...| Struthio aS

camelus ... : pre bhi dey 65-804?
Davainea (s.l.) casuarit, sp.n. ots .».| Casuarius picticollis e BuLs mn oka
# appendiculata, Fuhrm. ...| Unknown ... en at 130 Jeane
9 infrequens, spn... ...| Casuarius picticollis “yy. 260 21-34
ss fubrmannt, Southwell ...| Crocopus phoentcopterus ... 18 fe) | 5-301
ReeiRitinc (Ransomia) undulata, Fuhrm. ...| Corythaeola cristata re Rae 25-28
“ », campanulata, Fuhrm. | Perdix sp. ... pt: eee] 40-42 ie 27
” 55 vaganda (Baylis) __...| Haliaétus vocifer ... ...| Numerous 25 my
ah - about 1003 oe
(Paroniella) paradisea, Fubrm....| Manucodia chalybeata ...| about 2004 22 |b

s (Skrjabinia) oligacantha, Fuhrm. | Tynamus, sp. cic
rufescens ... at 34 21-2346

Davainea (s.l.) conopophilea, Johnston... 5 23

* According to Meggitt (1921) T. struthionts, as described by different authors, contains
more than one species, and reserving the name Davainea struthionis for the form firstly
mentioned (without proper description) by Houltuyn and described 1885 by Parona, he
separates from this latter the following species: D. linstowi Meggitt (1921) (= T. struthionis
of vy. Linstow (1893) and Hungerbiihler (1g10)) and D. beddardi Meggitt (1921) (= D. struthionis
of Zilluf (1912)). The size of the rostellar-hooks is stated to be different in all the three species.

1. According to v. Linstow (1893). 3. According to Fuhrmann (19009).
2. According to Fuhrmann (1920). 4. According to Skrjabin (1914).

The four suckers are rounded in size, and exhibit a well
pronounced musculature; they measure 0°4 mm. across; their border
is covered with very numerous small (10# to 13) hooks, which are
arranged in six to ten rows. A distinct neck occurs only in stretched
specimens. The segments, in most of my specimens, are much
broader than long. Gravid proglottides are, apart from extremely
contracted specimens, almost square.

ANATOMY.

As has been mentioned above, the worms are in part greatly
contracted, and their aspect is rather thick and compact. Such
conditions are to be found usually in worms which possess a well

§°

developed cortical parenchyma, subcuticular layer and cuticle. In
D. casuarii especially the first is rather wide and exhibits a well
marked longitudinal musculature. This latter consists of many
more or less distinctly separated bundles of various size. The largest
bundles are oval in shape, measuring about 40 to 54m. They are
composed of thirty-five to fifty fibres of various thicknesses.
Towards the subcuticular layer smaller bundles are scattered irregu-
larly, consisting of fewer fibres, or even of but a single one. The

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Fic. 2. Davainea casuarii, sp.n. Longitudinal section of two mature segments
Cp.—cirrus pouch; Ov.—ovary; Z.—testes; w/V.—ventral excretory vessel; /7.—vagina ;
Vd.—vas deferens; Vg.—vitelline gland. x 34.

transversal musculature separates very distinctly the medullary
parenchyma from the cortex. Fine dorso-ventral fibres are present
in both parenchyma layers, being especially well marked at the level
of the transverse excretory vessel. It is worthy. of note that rather
large calcareous bodies are scattered in the subcuticula as well as in
both parenchyma layers; they are mostly oval in shape, 10m to 164
in size, and deeply staining with haematoxylin.

Lxcretory system. The excretory system consists in main part
of a single pair of very large longitudinal vessels, which are
connected at the posterior border of each proglottis by a large
transverse canal. Although these longitudinal vessels run justly on
the transverse axis of the proglottides, having a diameter nearly
equal to the depth of the medullary parenchyma, there is no doubt
that they represent the ventral pair of the longitudinal vessels, for
in the anterior, mostly immature proglottides, I could undoubtedly
distinguish within the two large vessels two narrow, somewhat

dorsally located vessels without transverse commissures. These
dorsal vessels disappear apparently in the mature segments. The
wall of the excretory vessels is bordered by very minute rounded
cells, which seem to be parenchyma cells.

Genital organs. The openings of the genital ducts are
unilateral, the porus genitalis being situated about the centre of the
lateral border of the proglottides. A small atrium genitale is
present.

Fic. 3. Davainea casuarii, n.sp. Longitudinal section, showing the termination of
sex ducts. x 80.

Male organs. The testes are oval or spherical in shape and
67 to 81m in diameter. They occupy the whole free space of the
medulla at the sides of the female glands. On account of the
structure of the vas deferens and vagina, the testes are of course
more numerous on the antiporal medulla-half. Their total number
amounts to nearly fifty to sixty. In the younger, and also in mature
proglottides, the testes exhibit very interesting stages in the develop-
ment of the spermatozoa. These stages agree in many respects with
those described and drawn by Moniez (1881). The vas deferens is
a rather wide, coiled tube, the coils of which occupy dorso-ventrally
nearly the whole space of the poral medulla-half, displacing ventrally
the wide longitudinal excretory vessel just before entering the cirrus
pouch. After entering the cirrus pouch, the vas deferens forms a
rather large, coiled vesicula seminalis interna, which is usually filled
with spermatozoids. The cirrus is short (o'l mm.); it is surrounded
within the cirrus pouch by a dense network of very small cells,
representing, perhaps, prostate cells or merely parenchyma cells.

The thick-walled cirrus pouch is pyriform, and measures 0°25 mm.
in length by o'16 mm. in breadth; it does not extend beyond
the longitudinal nerve-stem, and thus does not reach at all the
longitudinal excretory vessel.

Female organs. The position of the vagina, 1.e., of the poral
portion of the vagina in proportion to the cirrus pouch, varies
according to the state of contraction of the strobila. In stretched
or normally contracted specimens it les immediately behind the
cirrus pouch; in extremely contracted worms, however, it is some-
times ventral, sometimes dorsal to the cirrus pouch. ‘The poral
third of the vagina, extending from the genital atrium just beyond
the poral longitudinal excretory vessel, is rather wide, darkly

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Fic. 4. Davainea casuarit, sp.n. Transverse section of a mature segment. Cp.—cirrus
pouch; Zm.—longitudinal muscles; Ov.—ovary; J.—testes; «w/V.—ventral excretory
vessel; V.—vagina; Vd.—vas deferens. X 42.

staining because of its rather muscular wall and especially on
account of the presence on its inner surface of very fine hairs. A
similar structure of the vagina is found in other worms, particularly
in some members of the family Davaineidae, in the Tetrabothriidae,
and also in certain species of the genera Tvichocephaloides,

Monopylidium, Octopetalum, etc. Just before the vagina opens
into the atrium genitale it bears a distinctly marked sphincter.
Within the poral longitudinal excretory vessel the vagina narrows
suddenly for a short distance and becomes then nearly as wide again
as the vas deferens, forming some coils before reaching the ovary ;
its course from the longitudinal excretory vessel to the ovary is
chiefly ventral, although it passes to the dorsal side of the
longitudinal excretory canal, as is usually the case. A distinct
receptaculum seminis is absent. The ovary is small; it 1s situated
in the middle of the proglottides lying in longitudinal sections
somewhat nearer to the posterior border of the segments and
consisting of fine lobes, which radiate in all directions from the
oviduct. Its diameter amounts nearly to one-fourth of the breadth

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Fic. 5. Davainea casuartt, spn. Transverse section of a gravid segment. xX 34.

of the medullary parenchyma. Dorso-ventrally it occupies the
entire depth of the medulla, being especially well developed on the
antiporal side. The vitelline gland lies quite dorsally in the median
part of the medulla, where it appears as a rounded compact organ,
measuring about o'o1 by 0°06 mm. A small but distinct shell-gland lies
on the main trunk of the oviductus. The uterus appears very early
as a rounded sac, which is situated ventrally in the middle part of
the medullary parenchyma. While enlarging it becomes apparently

divided into two oval parts, both becoming confluent soon after they
reach a larger size. At this stage the eggs segregate into groups
of three to four eggs, which latter eventually become egg-capsules ;
these are mostly oval or rounded in shape; they are bordered by
one to two rows of rounded, larger cells, while the two to four eggs
(in transverse sections there are visible mostly two, seldom three,
eggs in a capsule) are embedded in dense parenchyma containing
somewhat smaller cell-elements. The egg-capsules extend beyond
the longitudinal excretory vessels, and thus fill the whole
parenchyma. About one hundred and eighty capsules can be
counted in a transverse section; they measure 67” by 108 in
diameter.

Systematic comparisons.

As already mentioned above, there has been only one species
of the genus Davainea hitherto known from birds belonging to
the Casuariformes, viz., Davainea (s.1.) australis (Krabbe).
D. casuarii, sp.n., differs from this cestode in many respects, but
especially in the shape and size of the scolex and the rostellar-hooks.
Comparing this new cestode with other members of the genus
Davainea (s.1.), I find that it agrees in general with the type known
in Davainea. There is, however, no doubt that it bears some
characters which are to a certain degree rare or unusual in this
genus; such features are the considerable size of the rostellar-hooks,
the absence of one pair of excretory vessels and the well-developed
longitudinal musculature. Owing to these peculiarities it seems
that there exists a certain relationship between D. casuariz and the
genus Porogynia, Railliet et Henry (= Polycoelia, Fuhrm.). The
arrangement of the genital glands, however, which in our cestode
is of the usual type of Davainea, does not allow it to be assigned
to Porogynia, which latter, moreover, bears three rows of rostellar-
hooks on the scolex. On the other hand, it is not possible to place
this cestode into one of the genera recently established by Fuhrmann
(1920), mainly because of the above-mentioned unusual features.
Among all these new genera it is to the large genus Raillietina,
Fuhrm., sub-genus Raxsomia, Fuhrm., that our cestode should be
assigned, if we do not consider the above-mentioned characters to

be of systematic value, warranting the creation of a new genus or
sub-genus for it. For myself, I am inclined to believe that the
establishment of a new sub-genus might be justified.

The type specimen is in the Parasitological Museum of the Royal
Hungarian Veterinary College, Budapest.

DAVAINEA (s.1.) INFREQUENS, sp.n.
Host: Casuarius picticollts, Sclat.

Locality : Sattelberg.

Only a few specimens of this cestode were found in the same
host as D. casuariit, The worms are much ~ smaller and
narrower than the former species. Unfortunately there were only
incomplete individuals available. From two fragments, which
belong apparently to one,another, one can estimate the total length
of the strobila at about 80 mm. by a greatest breadth of 1°2 mm. in
the posterior third. The scolex is globular, measuring 0°5 mm.
across. It exhibits a fairly well developed rostellum of 0°25 mm. in
diameter, bearing a double row of typical hammer-shaped hooks.
Their number is about two hundred and sixty. They measure in
the anterior row 27m to 34m, in the posterior row 2Ip to 25m in
length. The suckers are spherical, their diameter being 0°13 mm.
They are bordered by four to six rows of hooks, 10m to 15m in
length. There is a well marked neck measuring about 2 mm. in
length. The proglottides are broader than long.

ANATOMY.

The internal anatomy of the worms exhibits the usual characters
of the genus Davainea; the structures seen in transverse and
longitudinal sections were much like those found, especially in some
Davainea species of Psittaciformes.

Excretory system. There is only one pair of large longitudinal
vessels, lying usually nearer to the ventral side.

Musculature. The longitudinal muscles are well developed;
they are composed of an internal layer consisting of about sixty

large oval bundles, and by a distinctly separated external layer,
which exhibits two rings of very small bundles consisting of at
most two to three fibres. Similar arrangement of the longitudinal
musculature occurs also in D. spiralis, Baczynska (1914).

Oval calcareous bodies are present, especially in the cortical
parenchyma.

Genital organs. It seems that the openings of the sex-ducts are
unilateral, lying on the left side; in one segment (of about twenty),
however, I found the opening on the right. The thick-walled cirrus
pouch is 0°18 to o'2 mm, 1n length, 0:06 mm. in breadth; it extends
to the poral longitudinal excretory vessel. The cirrus 1s short,
rather thick at its anterior end and covered with many spine-like
elements. It bears retractor muscles which radiate in all directions
to the wall of the cirrus pouch. Within this organ there is an oval
vesicula seminalis interna measuring 0°054 mm. in length. The
vas deferens forms many large coils in its course towards the middle
of the medulla. The testes are about nine to twelve in number,
lying not only at both sides of the female glands, but also in the
median line of the segments. They measure 0:05 mm. in diameter.

The vagina lies behind the cirrus pouch. Its structure is the
same as, e.g., in D. aruensis, Fuhrm. (1911) or in D. allomyodes,
Kotlan (1921). In the middle of the segments, shortly before
reaching the ovary, it forms a small spindle-shaped receptaculum
seminis. The bilobed ovary, when fully developed, is about o°2 mm.
in breadth; it lies in the middle of the segments. Behind the ovary
is situated the compact vitelline gland, which 1s about 0°08 mm.
broad.

Gravid proglottides are not available, and I am, therefore,
unable to give a complete description of this worm. The above
noted characters are, however, I believe, sufficient to distinguish this
form from other members of the genus Davainea, of which the
following must be considered mostly on account of the similar size
of the rostellar-hooks :—D. fuhrmanni, Southwell (1922), Raillietina
(Kansomia) undulata, Fuhrm. (1909), R. (R.) campanulata, Fuhrm.
(1909), and RX. (R.) vaganda, Baylis (1919).

The new worm in question seems in every way to be closely
related to D. fuhrmanni. Comparing, however, the characteristic
features of our worm with those of D. fuhrmanni, described in

detail by Southwell (1922), I conclude that there are some
differences which do not permit the two species to be united.

Such are :-—
1. The smaller number of the rostellar-hooks in D. fuhrmanni.
2. Larger and, in some respects, better preserved material

would perhaps show that the genital openings are irregu-
larly alternate in D. zxfrequens.

3. It seems that the cirrus pouch in D. znfreguens is longer and
rather narrower in size. ,

4. There is no mention in the description of D. /uhrmanni of
the distinct vesicula seminalis interna.

5. No mention is made of the presence in D. fuhrianni of
retractor muscles of the cirrus.

6. The prostate-cells surrounding the coils of the vas deferens
are inconspicuous in JD. infreguens.

7. Finally, it seems improbable that one and the same species
of worm should be found in Columbiform and Casuariform
birds.

The three other species mentioned above differ from D. infreguens
in the number of the rostellar-hooks and in other anatomical
characters.

The type specimen is in the Parasitological Museum of the Royal
Hungarian Veterinary College, Budapest.

REFERENCES
Baczynska, H. (1914). Etudes anatomiques et histologiques sur quelques nouvelles éspéces
de Cestodes d’oiseaux. Neuchatel.

Baytis, H. A. (1919). Some new Entozoa from Birds in Uganda. Ann. and Mag. Nat. Hist.,
merg, VoL Jit.

Fuurmann, O. (1909). Neue Davaineiden. Centralbl. f. Bact. u. Parasit., Vol. LXXIV.

— (1911). Vogelcestoden der Aru-Inseln. Abbandl. Senckenberg. Naturforsch. Ges.,

Vol. XXXIV. H. Merton, Ergebn. einer zool. Forschungsreise in den siidéstl.
Molukken.

(1920). Considérations générales sur les Davainea. Festschrift fiir Zschokke, No. a7.

KortAn, A. (1921). Vogel-Cestoden aus Neu-Guinea. I. Papagei-Cestoden. Annales
Muset Nat. Hungar., Vol. XVIII.

Krasse, H. (1869). Bidrag til Kundskab om Fuglenes Baendelorme.

Linstow, O. y. (1893). Zur Anatomie u. Entwicklungsgeschichte der Tanien. Arch.
mikrosk. Anatomie, Vol. XLII.

Mecertt, F. I. (1921). On two new Tapeworms from the Ostrich, with a key to the species
of Davainea. Parasitology, Vol. XIII.

Montez, R. (1881). Mémoires sur les Cestodes. Part I. Paris.

Soutuwett, T. (1922). Cestodes in the Collection of the Indian Museum. Am. Trop.
Med. and Parasit., Vol. XVI.

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59
AVIAN CESTODES FROM NEW GUINEA

III. CESTODES FROM GALLIFORMES
BY

Dr. ALEXANDER KOTLAN
From the Royal Hungarian Veterinary College, Budapest

(Received for publication 5 December, 1922)

Only one representative of the bird-group Galliformes has been
examined for parasites, viz., Megapodius brunneiventris, Mey.
The tapeworms which were found in the gut of this bird belong to
three distinct species of the genus Délepis, Weinl; one of these is
smaller and narrower than the two other species, and is, therefore,
easy to separate from these latter. It requires, however, a careful
examination to be able to distinguish the two other species, the
scolex and strobila of which are quite similar to one another. All
three species are, I believe, undescribed; the genus Dzlepis, so far
as I am aware, has not yet been recorded from Galliform birds.

DI PEPISSVCORKRE* spon:
Host: Megapodius brunneiventris, Mey.
Locality : Friedrich-Wilhelmshafen.

This is the smallest of the three species mentioned above; fully
matured specimens measure 15 to 20 mm. in length. The scolex is

Fic. 1 Dilepis yorkei, spn. Scolex. x 50.

very well developed, it is nearly as long (0'7 mm.) as broad
(o0°5 to o'6 mm.). It bears a rather powerful rostellum of conical
shape and of about 0o'5 mm. in length. This rostellum, when
retracted, is surrounded by a double muscular rostellar sac of about
o'7 mm. in length. On the anterior end of the rostellum there 1s a
button-like thickening of nearly 0'17 mm. in diameter, bearing fifty
to fifty-two large hooks, which are arranged in a double row.

Fic. 2. Hooks from the rostellum. A.-——-D. yorker; B.—D. leptopballus ;
C.—D. borvathi. x 230.
They measure in the anterior row 135m, in the posterior row
148m to 151m. The suckers are oval in shape and measure 0°42 to
0°44 by 0°25 to 0°30 mm. in diameter.

Behind the scolex there 1s a very short unsegmented portion,
which is usually broader than the segments of the anterior half of
the worm. The strobila of a fully developed specimen consists of
about one hundred and twenty to one hundred and fifty segments;
these are, as a rule, broader than long, except in macerated
specimens. Mature segments are 0'2 to 0°4 mm. in breadth and
0°05 to o'l mm. in length. ‘The greatest breadth (0°3 to o°5 mm.)
is attained in the last fourth of the strobila with gravid segments.

ANATOMY.

Body-wall and parenchyma. ‘The cuticle, as in other similarly
delicate cestodes, is rather thin and not at all compact. The sub-
cuticular cells are fairly well developed and arranged into two
or three rows. The body-parenchyma is of peculiar structure,
consisting of a loosely arranged reticulum with rather poorly
scattered cell-elements. Calcareous bodies were not found.

Musculature. The somewhat denser cortex is separated from
the very loose medulla by the longitudinal muscles, which are
arranged in two rings, each being composed of a row of small
inconspicuous muscle-bundles. Inside of the interior row there
is apparently a very poorly developed transverse musculature.
Dorso-ventral muscle-fibres were not seen.

Excretory system. In the anterior two-thirds of the strobila
there exist two longitudinal vessels on each side of the segments,
of which the ventral is slightly larger than the dorsal. In segments
in which the uterus reached a more considerable extent, only one
pair of longitudinal vessels can be seen. Transverse commissures
were not observed.

Fic. 3. D. yorkei, spn. Younger segment showing mature male organs and primordial
female glands. Pov.—primordial ovary ; Pog.—primordial vitelline gland ; Rs.—receptaculum
seminis; J.—testes. 170.

Genital organs. The first indication of the sex-organs appears
already in the first distinct segments. As in many other cestodes,
the male organs are markedly more advanced in development than
the female organs. The cirrus pouch attains its largest size by the
twentieth segment ; then follow the testes, which, however, disappear
about the sixtieth to seventieth segment, while the female glands,
which also appear rather early, reach full maturity after and
about the eightieth segment. Here also the uterus appears, and
grows rapidly to a considerable size.

The genital-openings are unilateral, being situated about the
middle of the lateral border of the proglottides.

Male organs. he cirrus pouch, compared with the size of the
proglottides, is a large tube of 0°18 to o'2 mm. length and o'o21I
to 0'°027 mm. greatest breadth. Its position varies according to the

state of contraction of the worm. In somewhat longer segments it
is directed obliquely to the anterior end of the segment. After
narrowing for a short distance it is continued by a very wide
vas deferens, which, forming one or two large coils, runs to the
posterior half of the segment. The cirrus seems to be a fairly
slender canal, which on its anterior end is apparently covered with
minute spines.

There are only four testes in each segment, situated in the
middle of the posterior third; they are 37m by 27m in diameter.

Female organs. The vagina, a fairly short and narrow canal,
runs dorsally to the cirrus pouch. It forms a large (about 0°08 mm.)
receptaculum seminis, which lies immediately within the dorsal
longitudinal musculature extending to, or but little beyond, the
middle of the proglottis. The ovary exhibits a peculiar structure,

Fic. 4. D. yorkei, spn. Older segment showing mature female organs. Cp.—crirus
pouch; Ov.—ovary; Rs.—receptaculum seminis; U.—uterus; Vg.—vittelline gland;
Vd.—vas deferens. "170.

which, though slightly modified, is characteristic of the two other
species also. In the present case it consists of four nearly equal
bodies, which are rounded or mostly oval in shape, measuring about
54m to 67" by 40u. One of these ovarial sacs is situated in the
poral half, while the three others lie antiporal, 1.e., two of them
ventral and one somewhat dorsal. Each ovarial sac sends out a
thin-walled canal; these unite into a larger, very short oviduct.
On the main trunk of the oviduct lies a rounded shell-gland.

A globular vitelline gland of 29” diameter is seen in the mid-line
towards the posterior margin of the segments.

The young uterus is a thin-walled sac, which lies ventrally in
the anterior half of the segments. Growing to a more considerable
size, its walls become more distinct; in this stage the female glands
disappear suddenly, the whole medulla being occupied by the
uterus. In the two or three last segments, however, the wall of the
uterus atrophies, the ripe ova filling up the whole space of the
proglottides. The rounded ova measure 54 in diameter.

I have named this species in honour of Prof. Warrington Yorke,
of the University of Liverpool.

The type specimen is in the Parasitological Museum of the
Royal Hungarian Veterinary College, Budapest.

DIEBLLS,. ERBOPRETAL Cl Seasp. 0.

Host: Megapodius brunnieventris, Mey.

Locality : Friedrich-Wilhelmshafen.

The longest worms, when fully developed, measure 80 mm. in
length. The scolex is rather similar to that of the former species,
its diameter being 0°68 mm. The rostellum is, in its main features,
hike that of D. yorkez, measuring 0°64 to 0°76 mm. in length. It
_ bears on its anterior, knob-like end fifty-two hooks arranged in a
double row, those in the anterior row being slightly smaller in size
(121m to 126m) than those in the posterior row (135m to 143,/).
The four suckers are rounded in shape, measuring 0°3 to 0°34 mm.
in diameter.

Segmentation begins just behind the scolex. The proglottides
are, as a rule, broader than long. The ratio of the length to the
breadth varies according to the different stages of contraction.
This ratio is in my specimens mostly as I : 4-6, so far as concerns
the proglottides of the anterior half of the strobila; backwards
(caudad) the length increases slightly, the ratio becoming as I : 3.
The anterior end of the proglottides is usually much narrower than
the posterior, which in most specimens shows a distinct thickening.

There is a very well pronounced overlapping, especially in the
posterior half of the strobila. Gravid segments are about 2°5 mm.
broad and 1°5 mm. long, and exhibit a considerable thickness.

ANATOMY.

Musculature. The longitudinal muscles consist of bundles,
which are arranged in two layers; the internal layer exhibits
twenty-six to thirty, the external sixty-six to seventy, mostly oval
bundles. The transversal musculature is very poorly developed;
mostly it seems entirely absent. The dorso-ventral muscle-fibres
are likewise but faintly distinguishable.

Fic. 5. D.leptopballus, sp.n. Longitudinal section of three mature segments.
Cp.—cirrus pouch; Ov.—ovary; Rs.—receptaculum seminis; T., testes; V.—vagina;
Vg.—vitelline gland. x 100.

It is worth noting that in none of the specimens examined could
any trace of calcareous bodies be detected.

Excretory system. This consists of two pairs of longitudinal
vessels, of which only the two wide ventral ones form a transverse
canal, this latter being approximately as wide as the main ventral
vessels.

Genital organs. The openings of the genital ducts are unilateral
and lie on the posterior third of the lateral border. The atrium
genitale is marked off by a dense network of very small cells of
rounded or oval shape.

Male organs. The cirrus pouch consists of a very long narrow
tube of about 0°68 mm. length and 0°02 mm. breadth. Its position
and course depend essentially on the state of contraction of the
proglottides, and especially on the progress in development of the
genital organs. According to this it runs in normally contracted
mature segments from the genital pore to the excretory vessels on
the transverse axis, and then turns dorsally, extending a little
beyond the middle of the segments. In longer proglottides
exhibiting fully developed genital glands, the course of the cirrus
pouch is very different from that described above, as (seen in optical
longitudinal section) it runs from about the genital pore to the
anterior third of the segment parallel to the lateral border, and
then turns abruptly to the median part. The cirrus pouch possesses
long retractor muscles, which extend beyond the antiporal ventral

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Fic. 6. D. leptophallus, sp.n. Transverse section of a mature segment. Cp.—cirrus
pouch; Dvm.—dorsoventral muscle fibres; Lm.—longitudinal muscles; Ov.—ovary;
Re.—retractor muscles of the cirrus pouch; T.—testes; d/V.—dorsal excretory vessel;
wW .—ventral excretory vessel; V’.—vagina. x 8o.

vessel. There is a coiled vas deferens, the coils of which lie in
transverse section mainly dorsal, and in a somewhat higher plane
than those of the vagina. The cirrus is a very long and slender
a] 3 . . . e e
(8) canal, forming usually many coils within the cirrus pouch; in
some specimens, in which it was extruded, I could observe that at

least on the anterior half it is covered with very minute spines.

Female organs. TYhe vagina is a strikingly long duct which
runs from the atrium genitale to the mid-part of the medulla, mainly
just within the internal layer of the dorsal longitudinal muscles ;
it forms very large coils, which fill dorso-ventrally the entire central
space of the medullary layer. The walls of the vagina are generally
thin and covered with rounded cells; immediately before entering
the small atrium genitale its walls present a slight sphincter-like
thickening. Just in front of the shell-gland there is a rather large
rounded receptaculum seminis. The ovary is composed of poral and
antiporal lobes, each consisting of distinctly separated groups of
acini, the poral lobe having about five groups and the antiporal
about ten. The acini are all rather similar in size, measuring about
43 across. Each group sends out a thin-walled, narrow canal, all
of which, running into a larger one, form a distinct ovarian bridge
connecting the two groups of the ovary; from the mid-part of this
bridge there arises, at first somewhat dorsally directed, a rather
wide oviduct. A similar structure of the ovary exists also in other
cestodes, of which the following may here be mentioned :—
Choanotaenia porosa (Rud.) (see Cohn, 1901), Ch. gongyla, Cohn
(1901), Anomotaenta platyrhyncha (Krabbe), A. microrhyncha
(Krabbe), and Ophryocotyle herodiae, Fuhrm. (1909). A compact,
somewhat bean-like vitelline gland lies in the middle of the medulla;
it measures O'14 mm. At the junction of the oviduct and vitelline
duct there is a distinct shell-gland of rounded shape.

The uterus appears at first as a rounded, thin-walled sac between
both groups of the ovary. It then grows very rapidly, sending out
oval diverticula laterally, and usually beyond the excretory vessels
as well. All of these sacs then flow together to form a larger one,
which at this point has already a more distinct cellular wall. It
is an interesting feature that the testes and the receptaculum seminis
still persist for a rather long time, the uterus having already
occupied transversely almost the whole of the proglottis. In the last

few segments the wall of the uterus atrophies, and they are entirely
occupied by the ova.

The ripe ova measure 64“ by 54m in
diameter.

The type specimen is in the Parasitological Museum of the
Royal Hungarian Veterinary College, Budapest.

67
DILEPIS HORVATHI, sp.n.

Host: Megapodius brunneiventris, Mey.
Locality: Friedrich-Wilhelmshafen.

Among the cestodes collected from this bird I found only a few
chains belonging to this new species. With the naked eye it is
not easy to distinguish them from D. leptophallus. “The worms are
apparently somewhat shorter than the latter species, the longest
specimens measuring 50 mm. ‘The scolex closely resembles in its
shape and size that of the former species, being 0°8 mm. in width.
The rostellum is still larger, and when retracted it extends with its
posterior end to 0°7 mm. behind the posterior border of the suckers.
On the anterior knob-like thickening there are fifty-two hooks
arranged in a double row; there is but little difference in size
between the hooks of the two rows. I found them to be gg in
length in the anterior, and 102m in the posterior row. The shape,
especially that of the anterior hooks, slightly differs from the type
shown in the two other species. The suckers are rounded in shape
and measure 0°3 mm. in diameter. There is no neck, except in
stretched specimens; the segmentation begins a short distance

Fic. 7. D. horvdthi, sp.n. Mature segment. C.—cirrus; Cp.—cirrus pouch;
Ov.—ovary; Rs.—receptaculum seminis; Sg.—shell gland; T.—testes; V.—vagina ;
Vd.—vas deferens ; Vg.—vitelline gland. x 80.

behind the suckers. The segments in my few chains are usually
broader than long, and show otherwise the same features as in
D. leptophallus. The greatest breadth found in the posterior part
of the strcbila is 1°5 mm.

Owing to the small number of specimens available belonging
to this species, I omitted to sacrifice a chain for the purpose of
cutting sections. The following description of the arrangement of
the sexual organs is, therefore, based mainly upon worms stained
as a whole with boraxcarmin and mounted in balsam.

The genital pores are unilateral, and lie about the middle of the
lateral border. It seems that the atrium genitale is of the same
extent as in D. leptophallus.

The cirrus pouch is a somewhat shorter but wider tube than it
is in the former species, measuring about 0°2 mm. in length and
0o'04 mm. in breadth; it is usually directed with its long axis
obliquely forwards. Within the cirrus pouch is found the rather
long cirrus, which at its extremity is distinctly thickened and
covered with minute spines. The vas deferens forms many coils,
which le mainly in the middle of the anterior end of the
proglottides. The testes lie behind the female organs; it seems
that they are less numerous (about fifteen to seventeen) than in the
former worm; they measure 54m to 64,4.

The vagina rises anterior to the cirrus pouch; it crosses the
posterior end of this organ and then forms apparently as many coils
as that of D. leptophallus. A rounded receptaculum seminis is
similarly present. |

The ovary exhibits the same peculiarities as in D. leptophallus.
If any difference exists in the structure of this organ in both forms,
it might perhaps le in the somewhat fewer number of the ovarian
lobes on both the poral and antiporal side.

The vitelline-gland is similar in shape and size to that of the
former species.

As gravid segments were not at hand, I am unable to give a
suitable description of the uterus and the ripe ova.

The main features which distinguish this species from
D. leptophallus are :—

1. The shape-and size of the rostellar-hooks.
2. The shape and size of the cirrus pouch.

I have named this species in honour of Dr. G. Horvath,
Director of the Zoological Department of the Hungarian National
Museum in Budapest.

The type specimen is in the Parasitological Museum of the
Royal Hungarian Veterinary College, Budapest.

Among the known representatives of the genus Dilefis, there 1s,
I believe, none which exhibits a closer resemblance, so far as the
above-mentioned peculiarities are concerned. The type of the
hooks might in some respects be likened to those of D. macrocephala,
Fuhrm. (1908), the scolex and rostellum of which are likewise
strong.

All the three species described above are closely related to each
other. This is proved by the foilowing features :—

1. The structure of the scolex and_ its integrate parts
(especially the rostellum and hooks).
2. The structure of the male organs, viz., the large cirrus
pouch, the reduced number of testes.
3. Lhe structure of the female organs in general and mainly
of the ovary.

In philogenetic respects 1t seems doubtiess that the three forms,
but particularly D. yorket, are very old representatives of the genus
Dilepis, and might be perhaps interpreted as a distinct group
within this genus.

REFERENCES

Coun, L. (1gor). Zur Anatomie und Systematik der Vogelcestoden. Nova acta, Abb. d.
Kais. Leop.-Car. deutsch. Acad. d. Naturforsch, Vol. LXXIX.

Fuurmann, O. (1908). Nouveaux Ténias d’Oiseaux. Revue Suisse de Zool., Vol. XVL.,
fasc. I.

——— (1909). Die Cestoden der Végel des weissen Nils. Results of the Swedish Zool. Exp.
to Egypt and the White Nile, 1901. No. 27. :

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AN ANOPHELES OF THE
MYZORHYNCHUS GROUP (Anopheles
amazonicus sp.N.) FROM SOUTH AMERICA

BY
Magee sek CURIS TORHERS, Clr I.M.5.
(Received for publication 4. January, 1923)
PLATE IV

Among some material at the Liverpool School of Tropical
Medicine, which owing to the kindness of Professor R. Newstead,
F.R.S., I was able to examine when home on leave, I was fortunate
enough to find a specimen of an anopheline brought by Dr. A. A.
Clark from the River Amazon, which not only seemed to be
new, but which appeared to be the first instance of an undoubted
Myzorhynchus, using this term in its restricted sense, recorded from
South America, or, indeed, from the New World. On looking up
material previously brought by Dr. Clark from this region,
Professor Newstead was able to find two other specimens of the same
species not quite in such good preservation. With Professor
Newstead’s kind permission, I give below a description of this
species under the name A. amazonicus. All three specimens were
females, the first mentioned being selected as the type and so
labelled in the Liverpool School collection.

A. amazonicus closely resembles the Old World species of the
group Myzorhynchus, and it possesses the ventral abdominal scale
tuft on the penultimate abdominal segment which Theobald used to
define the genus Myzorhynchus when he created it, though it is now
known that this character is *not present in all the species of the
group. In one character, however, A. amazonicus approaches the
Arribalzagia group, which normally, so to speak, represents
Myzorhynchus in South and Central America. The character
referred to is a kink or bend in the costa at the subcostal junction
associated with one or more small accessory dark spots in this

position. From the description it will be seen that A. amagzonicus
possesses this feature, though it shows nothing of the other more
salient Arribalzagia characters’ such as inflated wing scales, eye-
spots on the thorax, abdominal scaling, etc.

A very marked specific feature of A. amazonicus is the great
length of the anterior forked cell, which measures one-third of the
wing length and extends so far inwards that the bifurcation 1s at
the level of the junction of the subcosta with the costa.

Anopheles (Anopheles) amazonicus, sp.n.

DIAGNOSTIC POINTS.

An easily identified species characterised by :
(1) The wings with pale interruptions on the costa.

(2) The paips shaggy and without definite bands.

(3) The hind tarsi dark.

(4) The femora and tibiae unicolorous.

(5) The bifurcation of the second longitudinal vein at the

same level as the junction of the subcosta with the costa.

(6) A ventral scale tuft on the penultimate abdominal segment.

DETAILED DESCRIPTION.

Q. <A largish dark anopheline of Afyzorhynchus appearance ;
general coloration rather rusty black. Length of wing, 4°4 mm.

Antennae with the basal segment dark, free from scales; the
second segment with a small tuft of pale and darkish scales on inner
aspect; remaining segments free from scales. Palfz with the
segments, commencing with the rudimentary basal one, measuring
respectively 6, 21, 35°5, 21°5 and 16 per cent. of the whole organ.
Palpal index (measured from unmounted specimen) 0°7. General
appearance of palpi as in A. umbrosus, Theo. ; densely covered with
black erect or semi-erect scales almost to the apex, but with these
longer on the basal rudimentary and succeeding segment; apex
dark and organ without obvious pale bands, though one or two
light scales are present at the apices of segments three and four
difficult to see except in certain lights. Zadzuwm black scaled, the

scales somewhat erect over basal half; labellae darkish. Clypeus
dark, bare. Head with the frons and vertex with small very narrow
white scales, less prolonged than usual. The pale area smailer in
extent than usual; including the area of narrow scales in front and
about the same extent of the ordinary upright scales behind this.
Occiput with dark erect truncate scales of ordinary anopheline type,
extending below level of neck. Some broad white scales beneath,
between the eyes, gular chaetae black.

Prothoracic lobes with dense tufts of black erect scales and
numerous chaetae. Pvosternal hairs about four. Mesonotum of
uniform coloration, dull brown, the bare spaces, etc., not
conspicuous; chaetae inconspicuous and presence of median series
doubtful. The surface clothed with light coloured hairs, scantily
but fairly uniformly distributed over the dorsum, including the
fossae. Anterior promontory with a smallish area medially of long
curved, pale scales, not forming a conspicuous feature; laterally,
and extending about half way to the lateral angular process of the
mesonotum, are rather conspicuous erect pale spatulate scales.
Scutellum with about twenty-four large hairs and a second line of
two additional hairs on each lateral lobe; scattered smaller
impressions (scales or hairs) medially. Sfzracular hairs about two.
Pre-alar hairs about eight.

Wings with the length 444 mm. and the greatest breadth
1:05 mm. Base to subcostal junction 0°67, anterior forked cell
0°33, posterior forked cell o'19 of the length of the whole wing.
Forked cell index 1°8. The anterior forked cell unusually long, and
the bifurcation of the second longitudinal vein so far towards the
base of the wing that it is on a level with the junction of the sub-
costa with the costa.

The wing markings, as a whole, are rather diffuse, the pale areas
not being very distinct, whilst there is an admixture in places of
pale and dark scales. Costa mainly dark, but with the following
pale areas: a minute one near base, a well marked one at about
the junction of the inner with the middle third of the wing length;
a comparatively large one just internal to the subcostal junction,
a somewhat smaller one actually at the subcostal junction and one
at the apex of the wing not quite reaching to the point of junction
of the first longitudinal with the wing margin. The most

characteristic feature of the costal markings is the presence of the
small accessory dark spot, involving the costa only, which hes
between the two pale areas in the region of the subcostal junction.
In this position there is also seen a slight but distinct bend or kink
in the costa, as in species of Avribalzagia. The first vein is marked
as the costa, but with additional pale areas at the base and at the
accessory sector. The second vein has the stem mainly pale scaled,
with dark scales at the cross-vein and just distal to its origin; the
upper branch has a small pale spot just external to the middle, and
the lower branch an indistinct one somewhat internal to this; the
branches are also pale where they join the wing margin, though the
fringe itself here is dark. The ¢hivd vein is mainly dark, but has
light scaled areas, separated by a conspicuous small dark spot, on its
basal portion. The fourth vem stem is dark, with white scaled
areas near the base and proximal to the cross-vein. The branches
are mainly dark with pale scales forming one (or two) small spots
on the upper branch. The fifth vein has the stem with mixed dark
and pale scales, the anterior branch with pale patches near the base
and beyond the cross-vein, the posterior branch with pale (mixed
with dark) scales on its proximal and dark scales on its distal half.
The szxth vetx has alternate dark and light portions (four pale and
four dark areas). The /vimge is dark from the apical costal spot to
the space between the veins 3 and 4'1 where there is a light spot.
There is another somewhat indefinite pale spot between 4°2 and 5°1.
The remainder of the wing fringe is too rubbed in all the specimens
for description.

Except for the spots on the costa and on vein 2°1, the scales of
the under surface are all dark. The wing membrane is stained, but
is lighter at some of the pale scaled areas.

The scaling of the wing shows the normal arrangement, but the
truncated squames of the median series are very inconspicuous
owing to the development of the laterals, whilst these latter and the
plume scales of the reverse side of the veins approach each other in
character so closely that they are scarcely to be distinguished. The
general effect is a heavy scaling with rather uniform large obovate
scales. The squames show from seven to nine striations, the laterals
ten to eleven, and the plumes usually nine striations, but some
shghtly broader plumes are present on the fourth vein (upper

surface), where they may show as many as twelve striations. This
is the position where the large inflated scales of Avribalzagia occur.

The coxae, in the case of the anterior pair, have black scales
basally and anteriorly, also posteriorly and apically. The middle
and posterior pair appear devoid of scales. The anterior /vochanters
with black and white scales, the middle and posterior apparently
devoid of scales. emora of the anterior pair moderately dilated
in inner half. The femora and tibiae of all the legs without definite
markings, except that there is a lightish triangular spot on the mid-
tibia apically. Tarsal segments of all the legs dark, unicolorous,
but with the apices of segments one, two and three narrowly pale,
four and five being dark.

The abdomen with hairs only, except ventrally. Cerci with
hairs only. Ventral surface with hairs, except medially, where on
segments four to seven, somewhat nearer the posterior than the
anterior border of the segment, are small patches of white scales,
the number of scales increasing. up to. the patch on the seventh
segment. On the seventh segment, posterior to the white scales, is
a prominent projecting tuft of black scales.

HABITAT, etc. The specimens were collected by Dr. A. A,
Clark on his journeys up and down the Amazon. The type was
labelled ‘A. A. Clark, River Amazon, June, 1915.’

A. amazonicus is distinguished from A. vestitipennis, Dyar and
Knab, to which it has some resemblance, by the absence of speckling
of the femora and tibiae, by the tarsal markings and by differences
in the wing markings. It is distinguished from A. crucians, Wied.,
by the costal and palpal markings, from the Myzorhynchellas by the
uniform colour of the hind tarsi, and from A. peryassui, Dyar and
Knab, by the absence of eye-spots on the thorax, etc.

The only species about which some doubt must remain is
A. mattogrossensis, Lutz and Neiva. The description of A. matto-
Srossensis given by Lutz and Neiva (1911) corresponds in a number
of respects with the species now described. But the wings are
described as ‘rather dark, especially on the costa, where there are
two spots lighter in colour, greyish yellow; there is a band, whitish-
yellow, transversal and sub-apical, formed by a group of cream-
coloured scales ; there are others distributed in a somewhat irregular
manner upon the longitudinal veins, scarcely distinguishable by the

naked eye.’* As there are three quite distinct spots on the costa of
A. amazonicus, the correspondence here would not seem to hold
good. The description also says nothing about a ventral tuft on

the penultimate segment, though the ventral surface of the abdomen — |

is described as ‘having traces of elongate scales, narrow and rather
long.’t In A. amazonicus the scales, other than those forming the
tuft, are few in number, and would scarcely be described in the
words used in the description of A. mattogrossensis. Unfortunately
the description given of A. madtogrossensis is rather meagre, and
I am unaware of any other reference to this species giving any further
particulars, whilst the type is presumably in South America.

REFERENCE

Lurz and Netva (1gt1). Notas Dipterolojicas. Mem. Inst. Osw. Crux, Vol. I, p. 297.

*'The original passage reads, ‘ Azas bastante escuras, principalmente na costa, onde ha
duas manchas de cor mais clara, amarelo-pardacenta ; ha um risco branco-amarelado transversal
e subapical, formado por um agrupamento de escamas de cér creme; ha outras, distribuidas
de modo um tanto irregular, sobre as*nervuras lonjitudinais, que apenas se distinguem a olho
nu.

tT ‘na ventral ha vestijios de escamas alongadas, estreitas e pouco eompridas,’

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EXPLANATION OF PLATE IV

Fig. 1. Camera-lucida drawing of wing of A. amazonicus.

The
scales are not shown, but the positions of the dark and

pale scaled areas on the veins are indicated by shading.
ig. 2. Truncated squames of the median series (obverse scaling).
a. Costa internal to subcostal junction.

6, First longitudinal, basal portion.
Be do.

at level of subcostal junction.
ad. Stem of vein 5.

Lateral squames (obverse scaling).

a, 0; 6 21S i peewee

Scales zz situ on anterior branch of vein 5.
m. Median.

/. Lateral.
pl.

Plume scales of reverse side of vein seen through the
wing membrane.

Vein 5 is a normal vein, 72.é.,
the squame scales are uppermost.

Fig. 5. Plume scales of reverse aspect of veins.

a. Costa external to subcostal junction.
d. Stem of vein 5.

e. Stem of vein 4, upper surface. Vein 4 is a reverse
vein, 7.€., the squames are beneath and the plume

scales uppermost.

1 dey

Ventral view of terminal portion of abdomen, showing
scale tuft.

Jelb ees” 1H

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Gabe HTIOLOGY'-OF BEACKWATER
FEVER

Shas ded eee Od IN OTLISS
(From the Sir Alfred Lewis Jones Research Laboratory, Freetown)
(Received for publication 5 February, 1923)

Marchiafava and Bignami (1894), in referring to malarial
poisoning, say: ‘We may mention those morbid states which are
developed after the malarial (parasitic) infection has passed away ;
for instance, the post-malarial fevers, the delirium, the post-malarial
haemoglobinuria.’ Mannaberg (1894) emphasises the fact that
Kelsch and Kiener proved that in every severe case of malaria, even
in every malarial cachexia, haemoglobinuria may be observed, and
calls attention to the effects of lesions of the renal epithelium as
pointed out by Bignami. These observers recognised haemo-
globinuria as a relatively common complication or sequela of
malaria, and it is of interest that they recognised it as a fact that,
when haemoglobinuria develops after malaria, the parasites have
disappeared from the peripheral blood.

Later quinine was added as a supplementary cause of the attacks
of haemoglobinuria. Of the numerous more recent theories of the
cause of blackwater fever with which we are chiefly concerned here,
the first is Manson’s, who stated that blackwater fever is a disease by
itself, separate from and not dependent upon malaria; the second,
which is an advance on this. claims for blackwater fever that it is
produced by a living organism. Further suggestions, still largely
in the realm of speculation, have been made by which this or that
form of parasite is stated to be the cause of blackwater fever. Some
of these parasitic theories we shall have the opportunity of mentioning
later..

Although it is thirty years ago since Manson promulgated the
theory that blackwater fever is a disease entity, and although
numerous observers since have attributed to different organisms the

credit of being the cause of it, the specific parasite which gives rise
to blackwater fever is still undiscovered. The older malaria,
malaria-quinine and similar theories have suffered much at the hands
of critics, but to fill their place little of definite value has been
produced. The tendency has been to admit that while haemo-
elobinuria occurs as the result of malaria, and quinine and other
drugs, yet apart from these haemoglobinurias, and separable from
them even when, in the tropics, they occur in chronic malaria cases
who have been taking quinine, there is a definite recognisable
condition of haemoglobinuria which constitutes the main sign of
blackwater fever.

Castellani and Chalmers (1919) differentiate the haemoglobinurias
which may occur in the tropics into three groups: Symptomatic,
Toxic and Specific. Under symptomatic they put haemoglobinuria
occurring in the course of malaria, Raynaud’s disease, acute specific
fevers, and after severe burns. The toxic group includes haemo-
elobinuria resulting from the administration of quinine and its salts,
chlorate of potash, antipyrin, carbolic acid, and naphthol, or from
vegetable substances such as Vzcza faba. The specific group
comprises blackwater fever and paroxysmal haemoglobinuria. At
first glance such a classification appears of assistance to those who
are likely to come in contact clinically with blackwater fever cases,
eliminating from the sphere. of blackwater fever those confusing
elements which are introduced if what is in reality a symptomatic
or toxic haemoglobinuria is erroneously attributed to a specific
disease.

Before accepting this classification, however, it may be well to
consider in some detail the signs and symptoms by which these
varieties of haemoglobinuria are said to be distinguished from one
another. - In Table I, compiled from these authors, are given in
comparative columns the signs and symptoms under each form of
haemoglobinuria, the symptomatic group being represented by
malaria, the toxic by quinine, and the specific by blackwater fever.

Reviewing the table, it is worthy of note that the signs and
symptoms of quinine haemoglobinuria resemble those of an attack of
blackwater fever but are not so severe, and that jaundice is specially
mentioned as being slight or absent in the former condition. The
malaria group 1s allotted six positive signs and symptoms, which

TABLe I

Comparison of signs and symptoms of Tropical Haemoglobinurias.

Symptomatic

Haemoglobinuria in malaria,
Raynaud’s Disease, Acute
Specific Fevers and after
severe Burns

Toxic

Haemoglobinuria caused by
Quinine,
Potash, Antipyrin, Carbolic

Chlorate of,

Specific

Haemoglobinuria in Black-
water Fever and Paroxysmal
Haemoglobinuria

acid, Naphthol and Vicia

faba
Malaria Quinine Blackwater Fever.
1. Haemoglobinuria Resemble those of an attack 1. Haemoglobinuria
2. Fever of Blackwater Fever, but 2. Fever
3. Rigor are not so acute. 3. Rigor
4. Vomiting Jaundice slight or absent. 4. Vomiting
5. Prostration 5. Intense weakness
6. Anaemia 6. Anaemia
Additional.
(a) Anorexia
(6) Headache
Negative.—Rarity of severe (c) Pains Back and Legs
Jaundice (d) Nausea
(e) Diarrhoea}
(f) Thirst

| (g) Constipation
(b) Jaundice

(1) Hyperpyrexia
(7) Coma

Additional
signs and symptoms are enumerated under blackwater fever, and

also occur in the quinine and blackwater fever columns.

these evidently apply also to the quinine group, since the signs and
symptoms of the latter are said to resemble those of an attack of
The value of these additional
signs and symptoms as a means of distinguishing the blackwater and
quinine groups on the one hand from the malaria group on the other,
appears to be entirely discounted by the fact that these signs and
symptoms are all, without exception, adduced by the authors in their
foregoing description of one or other form of the malaria infections.

blackwater fever but are not so severe.

In the analysis of the differential diagnosis we find ourselves
reduced to the following :—
(1) In the malaria group, the rarity of severe jaundice.
(2) In the quinine group, the relative lack of severity of the
symptoms and the fact that jaundice is absent or slight.

Jaundice.

The patient whose chart is given below and who died of black-
water fever presented, some weeks before the date of the commence-
ment of the chart, slight jaundice, which passed off in a day; he had
a similar slight transient jaundice a week before his fatal attack of
blackwater fever. At the time of the second attack of mild jaundice
he had subtertian parasites in his blood in small numbers, and he
had also taken quinine irregularly. To what, then, are we to
attribute these mild attacks of jaundice? To haemolysis from
malaria, or quinine or blackwater fever? It appears legitimate to
assume that they were a manifestation of the same causes of
haemolysis as produced the marked attack of blackwater fever. It
is of importance to note that in this case during the fatal attack the
jaundice was not of an intense kind. Deep jaundice, again, is
known to occur in malaria; in fact, many authors include the ‘ yellow
fever-like type’ of malaria in their description. One of the cardinal
signs of this type 1s deep jaundice.

From a study of this table of differential aids, one must conclude
that although it may in the future be possible to distinguish
accurately between a malaria, a quinine and a blackwater haemo-
globinuria, this cannot by such aids be done to-day; the attempt at |
differential diagnoses on such slender evidence as the degree of
jaundice and the severity of the symptoms is unscientific. It is
commonly stated that blackwater fever is, owing to its severity, a
condition which leaves no doubt in the mind as to the diagnosis.
But if blackwater fever is a disease which presents itself in an acute
form, and in an acute form only, then it is, indeed, a disease
sui generis and incomparable with any other known disease.

Stephens’ views on blackwater fever are quoted by the authors:
‘Blackwater is not a disease fer se, but rather a condition of blood
in which quinine, other drugs, cold or even exertion, may produce a
sudden destruction of red cells. The condition 1s produced only by
malaria, and generally by repeated slight attacks, insufficiently
combated by quinine. In such cases of chronic malaria, 7.e., in those
suffering from anaemia, with repeated attacks of fever and repeated
doses of quinine, blackwater fever sooner or later almost certainly
supervenes, at least in tropical climates.’ The authors’ comment
upon Stephens’ account is as follows:—‘ These statements are too

sweeping if genuine blackwater is meant, otherwise the home of the
disease would be Ceylon, whereas it is so rare that we have never
heard of a genuine non-imported case; for in this island there are
Europeans and natives with just the conditions required by Stephens,
and yet they do not develop blackwater fever, because the only two
cases which we have met with or heard of in Ceylon in twelve years
were most probably cases of quinine haemoglobinuria. On the
other hand, Stephens’ remarks are correct if applied to quinine
haemoglobinuria.’ The last sentence of this criticism 1s important.
If it be a fact that in Ceylon there are Europeans and natives with
just the conditions required by Stephens, and if it be a fact that
Stephens’ remarks are correct if applied to quinine haemoglobinuria,
how are we to explain the low prevalence of quinine haemoglobinuria
in Ceylon, z.e., two cases in twelve years ?

The observation was made in this case of the occurrence of
transient jaundice on two occasions before the severe attack of
blackwater fever, malaria parasites being present on the second
occasion ; these preliminary attacks of jaundice may have represented
the occurrence in the blood of—in a less degree—the same changes
produced by the same cause as was active during the attack. The
probability of such mild haemolytic attacks is great and they are
easily overlooked by the patient, as they were in this case. It is
also unlikely that anything short of a severe attack will attract the
attention of the patient to his urine, and if the attention is not drawn
to the urine, it is certain that under the conditions of life in such
places as Africa a person will frequently fail to notice that his urine
is abnormal. In order to observe even considerable degrees of
haemoglobinuria, it is necessary to examine the urine in a suitable
vessel in a good light, precautions not usually possible for patients
living under the conditions which prevail in places where blackwater
fever occurs. I would suggest, then, that closer investigation will
reveal the fact that haemoglobinuria occurs frequently in the tropics
without being observed, and that still more frequently haemolysis
with slight jaundice occur without noticeable haemoglobinuria, and
that these conditions are in fact frequently due to the same causes
as blackwater fever and are mild forms of the same condition. Even
in England, one has seen a case who was walking about and was
unaware of the fact that he was passing haemoglobin in the urine in
quite noticeable quantity.

Short of ‘blackwater’ fever, which represents a_ gross
haemoglobinuria, there must be many degrees of haemolysis,
haemoglobinaemia and slight haemoglobinuria produced by exactly
the same agencies as produce ‘ blackwater.’ For such cases I would
suggest that the term ‘blackwater’ fever is not sufficiently com-
prehensive. We require a term for such conditions to indicate that
the process of haemolysis has not produced such a degree of
haemoglobinaemia as to result in the passage of haemoglobin in the
urine.

Numerous suggestions as to the nature of the causal parasite of
blackwater fever have been made during the last thirty years.
Protozoa, bacteria, spirochaetes and chlamydozoa are represented
among the suggested parasites. The suggestion of Sambon that
blackwater fever might be due to a piroplasma-like parasite has been
accepted by some, and there are many points of resemblance between
this condition in man and piroplasmosis in animals. Dudgeon
(1920) injected sterilized urine from cases of blackwater fever—
obtained during the period of haemoglobinuria—into animals,
without producing any ill-effects. This observer mentions as a
possibility that the disease may be caused by a filter passer.

Experimental Inoculation of Blackwater Fever Blood

In order to throw some light upon this important question of
whether or not there is a specific parasite or enzyme which causes
blackwater fever, an experimental inoculation was _ performed.
Blood was taken from the patient in the middle of what proved to
be a fatal attack of blackwater fever, and was injected into a healthy
European. ‘The blood was withdrawn from a vein in the arm into
a syringe containing citrated saline solution (2 per cent. Sod. cit.
and 0°85 per cent. Sod. chloride, equal parts) and was injected in
two portions into the recipient. The proportion of blood to citrate
saline solution was three to one, and of this about 10 minims was
injected deep into the region over the deltoid muscle at 3.45 p.m.
and 2 c.c. into the same region at 4 p.m. The recipient had
previously had malaria, the last infection being of the subtertian
variety, but he had been free from relapse for over eighteen months
and had taken no quinine for over eight months.

85.

There was no local or general reaction immediately following the
injections. Quinine bihydrochloride was administered orally in
order to obviate infection with malaria. The dates, times and doses
were as follows :—

December 4 30 IO grs. a3 4 p.m.
5 grs. =! 8 p.m.
ee 5 grs. aos IO p.m.
December § "ee 5 gts. wy 9 a.m.
5 gts. ay I p.m.

30 grs. in 21 hours.

It might be argued that the doses of quinine taken might be
capable of killing the parasite causing blackwater fever. Against
this we have the record of numerous cases of blackwater fever in
which even large doses of quinine failed to abate or ameliorate the
condition. Also in the fatal case in question, quinine was
administered by intramuscular injection on the sixth and seventh
days of the disease, 21 grains in all, without influencing the
temperature or improving the general condition.

Appended is the chart of the case giving the highest and lowest
temperatures recorded each day. It will be observed that at the
Days 1 2 3 a 5 6 7 8 9 10

Temperature Chart of fatal case of Blackwater Fever.

* Time at which injection was made.

time of injection the temperature of this patient was over 102°F.,
and that it remained high till his death five days later. It seems
likely that if there was an infective agent it should have been
present in the blood on the day of inoculation. Sources of fallacy
include the possibility that the parasite of blackwater fever is never
present in the blood at all; that it is present in such small numbers
that the amount injected did not include the organism; that the
parasite is present only for one or two days at the commencement
of the disease; that it has an unusually long incubation period; or
that the subject of inoculation was immune.

Result of inoculation

No immediate nor late effects were noted as the result of the
inoculation. Parasites were not found in the blood, nor was there
any rise of temperature nor haemoglobinuria observable during a
period of two months.

These facts appear to me to militate against the specific parasitic
theory of the etiology of blackwater fever.

SUMMARY AND CONCLUSIONS

1. The term ‘ Blackwater’ Fever, being applicable only to
conditions in which haemoglobin is present in visible quantity in
the urine, is too restricted.

2. The importance of pre- and post-haemoglobinuria states
which are inherent parts of the disease, is apt to be lost sight of
owing to the exclusive use of the term ‘ Blackwater’ Fever. Some
such term as ‘Occuit’ or ‘ Subliminal’ Blackwater Fever might be
used to express these conditions.

3. A differentiation of Tropical Haemoglobinurias into Malaria,
Quinine and specific Blackwater types is not possible merely on the
basis of the presence and degree of jaundice, or on the relative
severity of the signs or symptoms.

4. The existence of a parasitic cause of Blackwater Fever has
been frequently suggested ; an experimental human inoculation, with

blood from a severe case of Blackwater Fever which ended fatally,
elicited no evidence in favour of the existence of such a parasite after
an observation period of two months.

ACKNOWLEDGMENT

I have to thank Dr. J. Y. Wood, W.A.M.S., for performing the

inoculations.

REFERENCES

CasTELLANI and Cuatmers (1919). Manual of Tropical Medicine, 3rd ed.
DupcEon (1920). Fourn. of Hyg., Vol. XIX, p. 208.

Marcuiarava and Bicnami (1894). Monograph on Malaria and the Parasites of Malarial
Fevers. New Syd. Soc.

MannaberG (1894). Monograph on Malaria and the Parasites of Malarial Fevers. New Syd.
Soc.

Manson (1893). Trans. Epidemiol. Soc., New Ser., Vol. XII, p. 111.
STEPHENS (1907). In Osler’s System of Medicine, Vol. I.

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A NEW SPECIES AND A NEW VARIETY
OF CULEX FROM THE -BELGIAN
CONGO

BY,

A. M. EVANS
(Received for publication g February, 1923)

The new species and variety of Culex described in the present
paper occurred among two small collections of mosquitoes from the
Belgian Congo, which were sent by Dr. Severin to this School for
determination.

Culex mouchett, n. sp. (fig. 1).

This species is named in honour of its discoverer, Dr. Mouchet.

Head. Scales bordering the eyes white, flat below, and
becoming gradually narrower as they approach the vertex. Upright
forked scales golden yellow in front, very dark brown behind, inter-
mediate ones golden with very narrow black tips. Narrow curved
scales silvery white. Bristles projecting over vertex golden yellow.
Scales of proboscis, female palpi, and scales and hairs of male palpi
dark sepia.

Lhorax. Prothoracic lobes with whitish scales. Mesonotum
dark brown with bronzy scales, apparently discoloured by dampness,
and blackish bristles. Pale whitish scales on ante-scutellar space,
scutellum, and medially just behind the head. Pleurae with green
integument. Lower mesepimeron of type with one bristle socket
on left side.

Abdomen. ‘Tergites II to VII entirely covered with dark sepia
scales, except at side of tergites IV to VII, where basal triangular
patches of whitish scales may occur; these very small, except on
segment VII. Tergite VIII with large irregular patch of whitish
scales above. Stermites entirely whitish scaled.

Legs. Vestiture chiefly blackish-brown. Femora pale beneath;
middle tibiae and tarsi in some specimens with whitish scales
beneath, extending for the whole part of the length of the segments.

Wings. Plume scales on third vein ligulate, squames on costa
and first vein with six to eight striae. First fork cell slightly longer
than its petiole, its base distal to that of second fork cell.

Fic. 1. Culex moucheti, n.sp. Male hypopygium. 4A—phallosome, ventral aspect with
left parameral plate and part of basal plate; 1, 2, 3, and 4—processes of phallosome, /—left side.
B—left half of phallosome, postero-lateral aspect somewhat distorted, 1, 2, 3, and 4, as in 4.
C—tenth sternite, ventral aspect. D—lobe of side-piece.

Male hypopygium (fig. 1). Lobe of side-piece as in fig. 1, d.
The occurrence of only one moderately stout rod between the leaf
and the pair of very stout rods a constant feature in the five
specimens examined. Phallosome (Christophers, 1922) or mesosome

(Edwards, 1920) (fig. 1, a and 4) of simple structure, chitin of walls
thin and uniform except in region of basal tooth (4). Each half
of phallosome with two long terminal processes (1 and 2), the longer
weakly serrated distally, a short pointed process (3) projecting
dorso-laterally, and a chitinised tooth (4) at basal angle of dorsal
wall. Tenth sternites as in fig. I, ¢.

Meviie sc.,. six ‘cO-type GG, and three co-type 99 from
Stanleyville, Belgian Congo, 1922, Dr. Mouchet.*

This species is one of the pipiens group of Culex, as defined by
Edwards (1922). The hypopygial characters point to a relationship
with Culex pipiens, L., and C. ¢triflatus, Edwards, but the absence
of abdominal bands and of lines of white scales beneath the last two
male palpal segments would seem to indicate affinities with the
decens series of the group.

Culex annulioris var. congolensis, n. var.

Male resembling typical C. anznuliorts, Theo., in hypopygial
characters, banding of palpi, proboscis and tarsi, and scaling of
thorax, but with abdomen entirely dark scaled above, the median
basal and lateral apical white markings characteristic of anxnulioris
being entirely absent.

Type and co-type ¢ ¢ from Leopoldville, 1922, Dr. Duren.

REFERENCES

CurisTopHers, S. R. (1922). The development and structure of the terminal abdominal
segments and hypopygium of the mosquito, with observations on the homologies of
the terminal segments of the larva. Ind. ‘fourn. Med. Res., Vol. X, No. 2.

Epwarps, F. W. (1922). Mosquito Notes, III.p. Bull. Ent. Res., Vol. XIII, Pt. I.

* Since going to press two further ¢ of Culex moucheti have been received from Buta,
Belgian Congo, November, 1922, Dr. Mouchet.

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REPORT ON SLEEPING SICKNESS
IN EKET DISTRICT, SOUTHERN
NIGERIA

R. W. ORPEN

West African Medical Staff
(Received for publication September, 1922)

With the object of ascertaining the present position of sleeping
sickness in the Eket District, Southern Nigeria, a tour of the whole
district was made during April and May, 1922.

The route followed was from Oron to Awa, thus passing through
the centre of the Eket District. The main towns at which I stopped
and made enquiries were Oyubia, Ikorubo (site of old sleeping
sickness camp), Eket and Awa, whilst numerous small, so-called
villages were also inspected.

For the sake of clearness, and to avoid confusion, this report is
divided into three parts :—

(1) Result of cases recorded in the year 1912.

(2) Result of cases not recorded in 1912, but who gave
definite information that they had been inmates of the
camp.

(3) General enquiry into trypanosomiasis in the Eket District
during the months of April and May, 1922.

The whole investigation has been most difficult, the people
showing the greatest reluctance to impart any information on the
subject, and it was only after prolonged interviews, which were most
wearying, that eventually the information detailed below was
obtained. Very great assistance was given by the District Officer,
as well as by members of the Qua-Ibo Mission.

9+
PART I

In Macfe’s Report (1915) on sleeping sickness in the Eket
District in 1913, the following passage occurs :—

‘During the sixteen months in which sleeping sickness has been
under investigation, two hundred and twenty-two cases have been
identified in which the trypanosomes. have been demonstrated. In
addition one hundred and fourteen cases have been met with presenting
some of the clinical features of the disease, but in which parasites
have not actually been found. There can be little doubt that the
majority of these were cases of trypanosomiasis.’

The nominal roll of cases made by Macfie has unfortunately been
lost, but I have assumed that the two hundred and twenty-two cases
noted by him include eighty-nine definitely recorded cases, lists of
which, either in manuscript or typescript, were compiled in 1912 by
Foran and Gray, with a signed statement to the effect that all had
been diagnosed microscopically.

Of these eighty-nine cases I was able to trace thirty-five, details
of which are noted in Tables I and II. The remaining fifty-four
cases have not been traced and nothing is known about them.

Taste I.

Cases recorded in 1912 by Dr. Foran and Dr. Gray, and which are still alive.

| When
Name Age | Sex Residence recorded Remarks
Udo chee Cee ose M_ | Edem Idem 8.8.12 Seen April, 1922. Healthy. No
Ekpot glands, but thickening on sides
of neck which shows scars.

Usundurus 9 F Ikorubo 7.8.12 | Not seen. Is reported to be alive
and well.

Enoesiet ... ...| adult | M_ | Afaha Eket 8.8.12 | Seen 1922. Thickening on neck,
with scars; no glands to be
felt ; strong and healthy.

Obotnt Ekim...) 12 F Edem Idem 8.8.12 | Seen 1922. Strong and healthy ;

Ekpot no glands to be felt.

Etok Awa ...j adult | F Ikorubo 12.8.12 | Seen 1922. Healthy; no glands
to be felt. Slight scars on neck.

FO re mee F Idikpa 15.8.12 | Seen 1922. Healthy; no glands
to be felt. Slight scars on
neck,

TABLE I—continued

Name

Ekanem ...
Usoanwan

Umwa Etok
Amame ...
Mama .,..

Okposen ...

Adiah Ansudo
Akpanitauwen
Owoimaha

Esoena
Peter Nsooyo

Ntanwoo

Samuel Akpanuso

Eya

Ema

Wilson Akpan

Adiaha Esein

AG) §

.| adult

1 fe)

.| adult

adult

Residence

Tkorubo

Ikotesiokong

Ikorubo

Ikotesiokong

Ikotoquot

Ikorubo

Afaha Eket

Tkotesiokong

Ekpenobo

Efrieyo

Ikorubo

Effoe

Akai

Ikorubo

Inoiya

When
recorded

16.8.12

20.38.12

DIepel >

24.8.12

27.8.12

25.60.12

28.8.12

31.8.12

2.9.12

11-9212

13.9.12

24.9.12

28.5.12

Remarks

Not seen. Left the country
apparently well.

Seen 1922. Healthy. One small,
very hard gland on neck.

Seen 1922. Healthy; no glands
or scars.

Not seen. Alive and well, but
refused examination.

Seen 1922. Healthy; scars on
neck; three very small hard
glands to be felt.

Not seen. Said to be alive and
well.
Seen 1922. Healthy; one small

gland on neck freely moveable.

Seen 1922. Scars and thickening
of neck.

Seen 1912. No glands, but con-
siderable thickening both sides
of neck.

Seen 1922. No glands, but scars
both sides of neck. Strong and
healthy.

Seen 1922. Healthy; a few very
small hard glands felt. Gland

puncture refused.

Seen 1922. Healthy; a few small
glands to be felt; gland punc-
ture negative.

Seen 1922. Healthy; no glands ;
some thickening.

Seen 1922. Healthy; scars and
thickening, but no glands to be
felt.

Seen 1922. One small gland felt ;
gland puncture negative.

Seen 1922. Healthy; some scars
and thickening of neck,

Taste II.

Cases recorded in 1912 by Dr. Foran and Dr. Gray, and which have died.

In the twenty-three cases still alive, noted in Table I, the blood
was examined by wet and dry films, but in no case were trypano-
somes found. It must be noted, however, that Gallagher and Macfie
failed to discover trypanosomes in the blood of any of the cases,
diagnosis in every instance being made by gland puncture.

Very few glands were punctured during the present enquiry, as
in practically all cases the glands had resolved, or there was at most
an indefinite general thickening. In a few cases there were one or
two very small, extremely hard glands to be felt, fibrosis evidently
having taken place. Scarring of the neck to a variable degree was
common.

PART II

Apart from these thirty-five vecorded cases, I saw at Eket
twenty-eight wxzecorded cases whom I accepted as being former
patients at the camp from their history and from evidence, especially
from members of the Qua-Ibo Mission, who furnished me with

97
documentary proof of their personal knowledge of the individuals.
These cases were all in good condition, and did not exhibit enlarged
lymphatic glands or any other evidence of trypanosomiasis.

I was also informed of the deaths of seven uxrecorded cases who
were stated to have been inmates of the camp.

It is possible that the above-mentioned unrecorded cases are
included in Macfie’s one hundred and fourteen cases who presented
some of the clinical features of the disease, but in whom. parasites
were not actually found.

PART III

In endeavouring to ascertain if trypanosomiasis is still prevalent
in the Eket District, as many persons as possible were examined in
the towns and villages visited. As already stated, great difficulty
was experienced, as there appeared to be a very great reluctance to
impart information; in fact, so much so, that on several occasions it
was necessary to invoke the assistance of the authorities in order to
get the people of a particular place to come in for examination. At
no place was this more marked than at Ikorubo, the site of the former
sleeping sickness camp. I am informed that this was due to the
fact that the chiefs and headmen of the various surrounding villages
feared being called upon to furnish labour for another and new
sleeping sickness camp.

Twenty-three cases were seen which exhibited signs suggestive
of sleeping sickness, e.g., enlarged glands, but gland puncture and
blood examination were negative.

In all, one thousand eight hundred and six persons have been
examined by gland palpation, and the following table shows the
result. For purposes of comparison with the results recorded by
Macfie and Gallagher (1914), the same age-groups classification of
glands has been adhered to :—

+ = Glands obviously enlarged.
+ — = Sufficiently enlarged to be grasped.
+ —— = Enlarged, but not sufficiently to be grasped,

— = Normal.

Number of individuals of
each class examined wae 568 620 20 310 281

Taste III.

Sex | Male Female

Age |, erga) Saueaks Dpretiias- (O40 Sanh Ghote aia
+ 5 ( 0°88%) | 4 ( 0°64%)| 0 2 ( 064%) | 4 ( 142%) | ©
rar ae 40 ( 7°04%) | 7 ( 113%) ) 1( 50%) | 22 (71%) | 13 ( 4°63%) | ©
ects 372 (65°5%) [210 (33°87%) | 3 (1570%) [171 (55°16%) | 62 (22°06%) | ©

oe 151 (26°58%) 1399 (64°35%) | 16 (80°0%) [115 (37°19) [202 (71°88%.) | 7 (100.0)

As regards the above table, it will be seen that a large
proportion have glands that can be classified as + — — ; those with
4+ — are considerably smaller, whilst the + is a very small figure
to the total. The class —, or normal, equals 49°28 per cent.
of the total examined. In plain words, it is very rare to see
anyone who has enlarged glands to an extent that 1s noticeable.
Enlarged glands are certainly common, and they usually take the
form of small discrete, hard, shotty glands; in many cases they
can be very well compared with buckshot. In a few cases the
glands were observed to be suppurating, with a well marked sinus,
or they were distinctly soft; now and again the post-auricular ones
were enlarged, but this was comparatively rare.

Comparison of the above tables with those of Macfie and
Gallagher shows a striking reduction in the proportion of individuals
with enlarged glands.

The number of ‘ dirty heads’ was most marked, ranging from
a simple dry eczema to the most intensely deep punched-out necrotic
ulceration. There is no doubt that the ‘dirty head’ was more
common amongst males, and in a large majority of the heads
classified ‘ dirty’ the occipital glands were always involved.

Many hundreds of blood slides have been examined, both by
the wet and dry methods, but in no case was a trypanosome

found; similarly, gland punctures performed whenever possible were
invariably negative.

The incidence of enlarged posterior cervical glands among 1806 natives classified according to sex and age,

The people of Eket are of a very poor physique, and many of
them have the appearance of being ill-nourished and on the verge
of starvation. Various diseases

yaws, syphilis, rheumatism—
appear to be common. Eket is densely populated, the inhabitants
living in scattered houses, and not in any definite towns.

Very few biting flies were found, but a few tsetse were captured.

SUMMARY

Of the cases seen and referred to in the Annual Medical Report
for the year 1913, thirty-five cases have been traced out of a total
of eighty-nine recorded by Foran and Gray, and of these twenty-
three are alive and in good health (Table I).

In addition, twenty-eight cases have been traced who gave direct
information as to their having been in the camp, but whose
names were not recorded or cannot be traced. It is possible,
however, that some of these twenty-eight cases were amongst the
one hundred and fourteen noted by Macfie as presenting some
clinical signs of the disease but in whom the condition was not
diagnosed microscopically. In this connection it is gratifying to
be able to state that the names of the various medical officers who
had charge of the camp are well remembered, and on more than one
occasion the name of the doctor in charge was given without seeking.

Sleeping sickness has not been demonstrated during the recent
tour in Eket, although the cases selected for examination were chosen
as presenting clinical signs of possible trypanosome infection. The
natives themselves are of the opinion that the disease has died out,
and this statement is borne out by the members of the Qua-Ibo
Mission; at the same time this must not be accepted as a definite
statement, because, as already noted above, the natives are not
inclined to discuss the subject, and I am strongly of the opinion
that a number of possibly genuine cases were removed from villages
on my approach.

REFERENCES

Macrig (1915). Southern Nigeria Annual Report for the year ending 1913, pp. 56-62. Waterlow
and Sons, Ltd., London.

Macrre and Gatiacuer (1914). Ann. Trop. Med. & Parasit., Vol. VIII, p. 379.

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NOTES ON CULICIDAE IN VENEZUELA,
WITH DESCRIPTIONS OF NEW SPECIES

PART III

Aus M.) EVANS

(Recetved for publication 27 February, 1923)

Anopheles (Arribalzagia) punctimacula, D. and K.
Anopheles (Arribalzagia) venezuelae, Evans

Amongst the A77zbalzagia material from the Panama Canal Zone,
referred to in my previous paper (1922), were twenty-nine specimens
with at least one of the hind tarsi complete. Most of these
agreed with Howard, Dyer and Knab’s (1917) description of
A. punctimacula, D. and K., but two specimens had two hind tarsal
bands as in A. venezuelae, and two others showed a tendency to the
formation of the second band. Further, a very considerable amount
of variation was found among the specimens, with regard to the
spotting of the other segments of the tarsi, the left and right hind
legs of the same insect in one case being markedly different in
this respect. The tarsal characters used by- me to distinguish
A. veneguelae from A. punctimacula (1922, p. 217) are, therefore,
valueless.

I have also been able to examine numerous other examples of
A. venezuelae, kindly sent by Dr. Nufiez Tovar from Venezuela,
among them being seven specimens in which the last hind tarsal
segment has only one dark band. It was also found that the most
perfect specimens among these collections had a number, from two
to fourteen, dark squames scattered throughout the long pale
scaled area of the third vein, thus agreeing with the description of
A. punctimacula, D, and K,

102

A re-examination of the type of A. venxezuelae has revealed the
fact that one of the wings has several dark scales in this position.
I have, therefore, no hesitation in regarding A. venezuelae, Evans,
as synonymous with A. punctimacula, D. and K.

Culex maracayensis, 1. sp. (fig. 1)
MALE.

Proboscis with a narrow whitish band on outer third. Palpz
with scales mostly dark brown, pale scales creamy, forming a narrow
band on basal half and a wide band on proximal half of long
segment, bases and apices of all the segments pale scaled. Occzput
with silvery narrow curved scales in front, brassy ones behind,
upright forked scales blackish. Prothoracic lobes with whitish
narrow curved scales and pale brown bristles. Integument of
mesonotum reddish brown. Dorsum with two broad bare stripes,
narrowing distally. Vestiture of rather sparsely distributed golden
brown and silvery scales, the latter occurring chiefly at anterior
lateral margins, on anterior fourth of median area, around antt-
scutellar space, and in two small oval areas on posterior half of disc.
Bristles, numerous, brown.

Abdomen. ‘Tergites dark brown scaled with narrow irregular
basal bands of whitish scales. Sternites clothed with transparent
whitish scales.

Wings with dark brown scales. Bases of fork cells about equ1-
distant from base of wing. First fork cell about twice as long as
its petiole.

Legs. Femora pale beneath, the pale area being very well
defined on the femur. Apices of front and mid femora narrowly
pale. Front tibia with conspicuous apical white patch above, about
twice as long as the average width of the tibia in dorsal aspect.
Mid tibia with very small pale apical spot, pale scaled beneath
throughout, hind tibia with a well defined stripe of creamy scales
extending along most of its length dorsally and a well defined apical
white ring. Front and mid tarsi with first two segments narrowly
pale apically, other segments of front tarsi without white, those of
mid tarsi with one or two pale scales apically. Hind tarsus, with
conspicuous pale rings apically and basally on all the segments.

103

Hypopygium (fig. 1). Side-pieces (A) with clasp narrowing
gradually towards distal extremity, articulated spine narrow. Lobe
of side-pieces (B) an undivided, distally directed arm, bearing three
stout rods, of which two are sub-equal, and longer and stouter than

0-1 Millimeter

—————|
O 2 Millimeler-

OO5Millrmeler ~° «eee 4

Fic. 1. Culex maracayensis, n.sp. Hypopygium. A.—Side piece. B.—Lobe of side
piece. C.—Phallosome, ventral aspect, semi-diagramatic, to same scale as 4; 1, 2, 3 lobes
numbered as in D.; t.—position of teeth. D.—Half of phallosome under pressure, ventro-
lateral aspect ; ~.—process of lobe 2; t.—teeth on upper side of lobe 2. £.—Tenth segment,

dorsal aspect.

the third. Phallosome (C & D) with each half divided distally into
three lobes; inner lobe (1) ventral, with chitin distally thrown into
ridges, which give rise to a denticulate appearance, particularly at
the margin. Second lobe when flattened out appearing as a thin

104

plate with an external pointed process (p.) and bearing on its dorsal
surface a row of blackish chitinous teeth, five large and three or
more small (some of these teeth are largely obscured by the others
in the figure); four of the large teeth continued proximally as thin
chitinous ribs. Third lobe (3) arising dorsally to second lobe,
elongate, curved and much narrower than the first and second lobes.
Tenth segment a membraneous lobe with curved basal arms, distal
margin spinose, tergal surface with paired chitinous plates, a spinous
area distally, and a group of four setae and a row of seven setae
laterally.

Length, c. 400 mm, Wing, c. 3°0 mm.

Type: One ¢, Maracay, October, 1922; Dr. Nufiez Tovar.

This species appears to be most closely related to C. coronator,
which it resembles in colouration and in the character of the tenth
sternites.

Culex paganus, n. sp.
MALE.

Palpi very short, as short as those of female. Head: Antennae
plumose, hairs brown; proboscis dark brown scaled, expanded
apically ; eyes black, occzput black clothed with white, narrow curved
scales, white flat ones at sides below, and pale yellowish brown
upright forked scales. Clypeus yellowish brown, sub-globular.

Prothoracic lobes whitish scaled. Integument of mesonotum
pale olivaceous, darker where sub-median bare stripes occur and in
posterior lateral areas. Scales whitish and pale yellowish brown,
the whitish ones predominating anteriorly and at sides. Bristles
long, dark brown. Pleurae pale green.

Abdomen with grey integument. Scales of tergites dorsally
very dark brown with sub-metallic bluish lustre, ventrally whitish
with bluish lustre. Sternites whitish scaled.

Legs unbanded, vestiture dark sepia, femora pale beneath.

Wing. Scales of costa and sub-costa dark sepia, on other veins
semi-transparent with obscure bluish tinge in certain lights. First
fork cell almost three times as long as its petiole, second twice as
long as its petiole.

Hypopygium. The main features are illustrated in figure 2.
Tenth sternites slender, comb-shaped distally with about six teeth.

105

FEMALE,

Antennae pilose, hairs brown. Occiput with creamy narrow
curved scales and pale straw-coloured upright forked ones.
Mesonotum with integument uniformly brown, pale scales almost
confined to edges of disc and lateral depressed areas.

O-1 Mituime+ser.

Fic. 2. Culex paganus, n.sp. Hypopygium. .4.—Apical part of side piece, general setac
of vestiture omitted. B.—Half of phallosome, lateral aspect ; 4.6.—basal hooks. C.—Trans-
parent triangular plate.

Legs. Hind tibia and metatarsus with a few brassy scales
beneath.

Type: ¢ and @ from villages, Estada Aragua, Venezuela,
23rd August, 1922; Dr. M. Nufiez Tovar.

This species apparently approaches near to Culex (/sotomyia)
bifoliata, Dyar, from the Panama Canal Zone, in the structure of
the male hypopygium. The leaves on the stem of the upper division
of the side-piece are, however, described as ‘ crooked curved leaves,’
and although the leaves (/.) in C. paganus are apt to be folded in
mounting, they could not appear crooked unless greatly distorted
in this process. There are also a number of other differences in

106

points of detail, but in the absence of a figure of the structures in
C. bifoliata it is difficult to estimate the value of these. In vestiture,
however, C. paganus differs very greatly from the Panama Canal
species, in which the upright forked scales of the head are white,
the vestiture of the mesonotum consists of ‘ fine dark brown hairs,
and the abdomen is entirely black.’ There can be no doubt,
therefore, that C. paganus is specifically distinct from C. (/sotomyiza)
bifoliata, Dyar.

Culex (Neomelanoconion) chrysothorax, Newst. and Thomas

I am now able to confirm the occurrence of this species in
Venezuela, which has hitherto rested on the record of a single female
collected by Professor Stephens at Mene Grande. Iwo males
and two more females were taken at Maracay, 5th October, 1922,
by Dr. M. Nufiez Tovar.

Psorophora tovart, Rear (figs. 3 and 4)

A considerable amount of material of this species has been
received since the publication of its description (1922), which enables
me to give a comprehensive account of the thoracic and abdominal
colouration, as well as a description of the male.

FEMALE.

Mesonotum. The distribution of scales of different shapes and
colours is illustrated in figure 3. The narrow curved, spindle-
shaped, and smaller broad curved scales (fig. 3, C, D & E), which
are usually dull brown or yellowish brown, are in some specimens
dull pale yellow and whitish. The very broad, much curved scales
(B, Bi) are usually pale creamy yellow, sometimes pale yellow.

Abdomen. The broad, pale yellow, apical, dorsal bands which
are complete on segments two to six of the type, may be interrupted
medially by dark scales on segments three to six, four to six, five
to six, or six; or they may be separated from the posterior margins
medially by a relatively small or large dark scaled triangular patch
on these. segments.

107

OC 7 Ae
Vo Ci VFS
(e) J cy yy
SUZ Bf
an t
lp

G3
Ygyeg iT Gy

\ \
yy J \
by:

9)
Q0 O70
9

—_———o
0-5 Miltimeler

Fic. 3. Psorophora tovari, Evans. A.—Mesonotum of female. B., C., D., and E.—Scales
from regions b., ¢., d., and ¢., of mesonotum, mounted in balsaam; B 1.—scales from region b.
as seen in sit.

MALE

Palpi entirely clothed with blackish scales with deep metallic
blue reflections and black hairs, last two segments incrassate.
Hairs of antennae brown, tori shining black. Occiput, mesonotum,
legs and wings as in female.

Abdomen with apical bands usually complete on segment two,
complete or divided on segment three, and generally interrupted
medially either partially or compietely on the other segments.

Hypopygium (fig. 4). Claspettes (harpagones) with nine (this
number may be subject to slight variation) stout filaments (/.)
arising from prominences along distal border, and a row of about
sixteen to twenty very delicate setae with distal portions swollen
and produced into fine filamentous processes as shown in the figure.

Type: do and nine co-type $d ‘from Maracay Region,
Venezuela, 1922; Dr. M. Nufiez Tovar. Co-type 9 from Maracay,

108

10th October, 1922, and others trom Maracay Region, July, 1922,
292 14; Maracay, 5th June, 1922, 9 9 30; San Meteo, 2nd June,
1922, 9 9/27; Guacara, I922,.9 9 4; Lagua, 15th June; 1922)
292. Dr. M. Nufiez Tovar.

0-05 Mucimeter

Fic. 4. Psorophora tovari, Evans. Apical portion of claspette, ventral aspect ; f.—stout
filament ; 4.—expanded hair.

This species is evidently closely allied to P. cyanescens, Coq., and
P. purpurascens, Eds., specimens occurring which resemble one or
other of these species in abdominal markings. The three species
appear to differ chiefly in mesonotal vestiture; P. cyanescens having
‘broad soiied silvery scales intermixed with some narrower brown
ones . . . . especially on centre of disc, but not forming any
defined pattern’ (H., D. and K., 1915), while P. purpurascens,
Eds., has the mesonotum with ‘ flat silvery grey scales, darker, but
not conspicuously so, in the centre of the mesonotum.’

Psorophora ciliata, Fab.

In a previous paper (1922) I recorded the occurrence of two
specimens of this species near Maracay, and Dr. Nufiez Tovar has
subsequently sent further material from this region. In view of

109g

Dyar’s recent study of the species of the czlzata group of Psorophora,
; and their distribution, and also of the fact that they exhibit
considerable differences in thoracic pattern from P. calzata, a further
: discussion of the Venezuelan specimens is necessary.

Dyar recognises four species of the célzata group of Psorophora
in the Argentine region, and states that, apart from Theobald’s
record of it in British Honduras, true célzata has not been recorded

) south of Tampico, Mexico. Further, he separated P. /zbzalis, a
| South American species, from czlzata by the slight differences of
| mesonotal pattern together with the markedly discontinuous
| distribution. Now, in none of the Venezuelan specimens does the
mesonotal pattern conform exactly to that of P. czlzata, and in some
cases (fig. 5, A and B) it differs quite as much as that of P. ¢zdzalzs,

i x
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ee = ; ( f- ;
1 MILLIMETER rae . broad flat scales.

PIC 5, Psorophora ciliata from Venezuela. Mesonotal patterns of three different specimens. A. and B.—Female ; C.—Male.

110

resembling this latter species in the reduction of the long lines of
black scales (fig. 5, C, 6/.). All the five specimens differ from
P. ciliata in having the median line of narrow curved scales not
golden in the antescutellar space. In three of the specimens these
scales are entirely silvery (fig. 5, A & C), while in the other two
(fig. 5, B) they are mostly pale yellow. Owing to the amount of
variation which exists among only five specimens from this region,
and the proximity of Venezuela to Central America, I regard these
specimens as specifically identical with P. czlzata.

Megarhinus trinidadensis, D. and K.

Males, larvae and pupae agreeing with the description of this
species, and females differing in the absence or reduction of white
on the third mid-tarsal segment, have been received from
Dr. M. Nufiez Tovar. This difference does not seem to justify the
separation of these specimens from the Trinidad species.

Bred in laboratory, Maracay, 1st November, 1922, Dr. M. Nufiez
Tovar, ¢ 1, 2 1; Mariara, Est. Aragua, 11th September, 1922,
3 1; Maracay, 4th June, 1922, d 1; Maracay region, $1, 2? 2 20.

Goeldia longipes, Fab.

Five females taken at Tucupido, December, 1922; Furmero,
8th June, 1922, 9 9 2; and Maracay region, June and July, 1922,
2 2 2, by Dr. M. Nufez Tovar, are referred to this species, although
they differ slightly from Howard, Dyar and Knab’s (1915) account
of it. The mesonotal scales have a distinct sub-metallic blue colour,
when the thorax is viewed from behind, and the scales on the scutellum
and ante-scutellar space are peacock-blue and greenish-blue. In
these respects they resemble L. cudicivora, D. and K., but they differ
from this species, and resemble G. longipes, Fab., in the ciliation
and colouration of the hind legs. The female palpi are said to be
equal in length to six antennal segments in G. longipes, and to four
in G. culictvora; in the Venezuelan specimens the female palpi equal

nearly five antennal segments, that portion projecting beyond the
clypeus being equal to four segments.

111
Wveomyta (Decamia) pseudopecten, D. and K.

A male specimen taken at Maracay, 2nd September, 1922, by
Dr. M. Nufiez Tovar, was found to agree closely with this species
in hypopygial characters, but the long paired hairs of the side-piece,
though longer than the clasper were less than twice its length.

REFERENCES

Dyar (1922). The species of Psorophora of the ciliata group. Ins. Ins. Mens., Vol. X, Nos. 7-9.

(1922). Notes on Tropical American Mosquitoes. Jns. Ins. Mens., Vol. X,
Nos. 12-12.

Epwarps (1922). Mosquito Notes, III. Bull. Ent. Res., Vol. XIII, Pt. 7, pp. 1-128.

Evans (1921). Notes on Culicidae in Venezuela, with description of New Species, Part I.
Ann. Trop. Med. & Parasit., Vol. XV, No. 4.

— (1922). Part II. Ann. Trop. Med. & Parasit., Vol. XVI, No. 2.

Howarp, Dyar and Knap (1915-1917). The Mosquitoes of North and Central America and
the West Indies. Vols. III and IV. Washington.

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the death of

H.R.H. Princess Christian

Princess of Great Britain and Ireland

Honorary President of the Liverpool
School of Tropical Medicine

from 1905

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113

A REVISION OF THE AMPAHISTOMATA

OF MAMMALS

Pp. A. MAPLESTONE

(Received for publication 12 January, 1922)

Pirates V-VIII

TABLE OF CONTENTS

PAGE
Amphistomata... af ide SPST TBS Gastrodiscus ve
Balanorchis ss ‘isi ei poner Oe . o aegyptiacus ...
2 anastrophus ... | ee EGS fh secundus
Brumptia bie - act ... 196 Gastrothylacidae
+ gigas ... Ree vee et OG Gastrothylax :
Carmyerius s Te mor 78 ; i crumenifer ...
- cruciformis ... sgt aiot Homalogaster
Peat) 5, exoporus ae ras st 184 ~ paloniae
be os gregarius ay er Tin s4 Paramphistomidae...
ee, Spatlosus Sc ena A agg? Paramphistominae...
* Wwenyoni Ese ee aay LOE Paramphistomum ... eh
MprorcPis ...° ... Sane sas Si LOS 3 buxtfrons
is fabaceus or sale Sem as 5 *, cerv!
Cladorchinae a am se Lien 5S . a explanatum
Cladorchis ... bi a. Aa) wh. #106 ~ gigantocotyle
» ~~ asper 167 x liorchis ...
» giganteus 167 : _ orthocoelium
» — —pyriformis _ Sauk 166 “ , pisum
9» Ssubtriquetrus ... es ae OF, 4 wagandt
Cotylophoron on re seer lis) Pfenderius ... —
cotylophor um are a: » —— papillatu
ie minutum ... te ar se Pseudodiscus
Diplodiscus x oe adh oy iG : 3 collinst
Fischsederius as as Ben meets bis ss hawkestt
‘ ~ elongatus ..3 : obey (2776 ¥ watsont
9 cobbold1... nay “pe eh G25 Stephanopharynginae
Gastrodiscidae... es a. ftth 487 Stephanopharynx ... oe
MetCodeS) 8, ck nae, 198 i 9 compactus
> hominis... Fas Ad eine Taxorchis ... aa
3 schistocotyle

* Species in the Museum, Liverpool School of Tropical Medicine.

114

INTRODUCTION

The publication by Stiles and Goldberger (1910) of their monograph
on the“ PARAMPHISTOMOIDEA ° called forth hostile criticism from many
writers, notably Braun (1911), Odhner (1911), and Looss (1912). The
chief objection of these authors to Stiles and Goldberger’s classification is
that they ignored the recent attempts that had been made to classify the
Trematodes by using affinities in the lymphatic and excretory systems
as a basis. Although not agreeing with the classification of Stiles and
Goldberger, none of the above writers formulated an alternative system,
and although Looss in his article of 1912 says he has been several years
trying to group the Amphistomes on the comparative: features of the
lymphatic, excretory and copulatory systems, this work has not been
published yet, so far as the writer is aware. The result is that the confusion
caused by Stiles and Goldberger’s multiplication of genera, etc., still
exists. Stunkard (1917), in a resumé of the group, makes a “‘ provisional ’
attempt at reclassification, but as he did not examine any material except
three or four species from American fish, for which he makes two new |
genera, he is unable to discuss whether many of the existing species and
genera are valid or not. All that he has done is to remove Stiles and
Goldberger’s family names, leaving some of their sub-family and their
generic names ; to add a sub-family of Looss containing one of his own
genera, one of Looss’, one ‘ (Gen. nov.) spinulosum,’ and ‘ Genera of
uncertain position ’ ; and to recommend a new sub-family for Balanorchis
and another sub-family for his own genus Zygocotyle. Such a procedure
cannot be regarded as improving matters.

It is probable that Looss’ suggestion to classify the members of the
group on the minute anatomy of the lymphatic, excretory, and copulatory
systems is sound, but it appears to be too complicated for practical
purposes.

In view of the opportunity presented by the large collection of
Amphistomes in the Museum of the Liverpool School of Tropical Medicine,
the writer decided to undertake a revision of the group with the object
of providing a working classification. This is not based on minute
histological study but on easily ascertained anatomical characters, and
some of the divisions used by Stiles and Goldberger (e.g., the three families
Gastrodiscidae, Paramphistomidae, and Gastrothylacidae) have been retained
because they serve to divide the group on easily distinguished external

II5

characters, and are hence of practical use. It is realised that the present
attempt at classification does not take cognizance of the standards required
by the more advanced systematists such as Looss and Odhner, but it is
claimed for this system that it is reasonably simple and consistent, and,
so far as the Amphistomes of mammals are concerned, enables a species
to be determined with considerable accuracy.

Looss also draws attention to the fact that Stiles and Goldberger
failed to make any allowance for variation due to difference in age or the
state of contraction of specimens, and have made many species on unsound
data in consequence. Throughout the present work these factors have
been investigated as fully as possible, and an attempt has been made to
show to what extent they influence the appearance of the several species
of the group. In their monographs Fischoeder and Stiles and Goldberger
both deal so fully with the synonymy, that it has not been considered
worth while to go back beyond these authors.

Note.—The reader is referred to Cohn (1904), Daday (1907), Looss
(1912), and Stunkard (1917), for the most recent information regarding
Amphistomes parasitic in hosts other than mammals.

AMPHISTOMATA, Rudolphi, 1801, e.p., Nitsch, 1819

Defimtion.—Digenea : two suckers, the anterior surrounding the
mouth and the posterior terminal or ventro-terminal behind the genitalia ;
gut forked; excretory pore opening dorsally towards the hinder end ;
testes generally in front of ovary; almost always thick worms more or
less circular in section.

Key To FAamI.igs

1. Body usually flattened and divided into anterior and posterior portions ; ventral

pouch absent ... ey Ge a ws nds ant Gastrodiscidae

Body usually conical and not divided into anterior and posterior portions 2

2. Ventral pouch absent... Lee i ae at. Parampbhistomidae
Ventral pouch present ... is at ¥ a3 ... Gastrothylacidae

Family PARAMPHISTOMIDAE, Fischoeder, 190r.

Definition.—Amphistomata : body not divided into two portions ;
ventral pouch absent.

116

Key To SUB-FAMILIES

1. Oral diverticula absent ... + oe: Ane fo Paramphistominae
Oral diverticula present ... = ~b Tt pe: in’ ae sts
2. Oral diverticula double ... ts re tied og 1 Cladorchinae*
Oral diverticula single .. a ae Sas ot Stephanopharynginae

Sub-family PARAMPHISTOMINAE (Fischoeder, 1901), s.str., Stiles and
Goldberger, I9Io.

Definition.—Paramphistomidae, without oral diverticula.

Kry To GENERA

Genital sucker absent aes Bes We he At ... Paramphistomum
Genital sucker present ae io ite 7.) nerd cb: Cotylophoron

Genus Paramphistomum (Fischoeder, Igor), s.str., Stiles and Goldberger,
IgIO.

Definition.—Paramphistominae, without a genital sucker.

Type species Paramp/lustomum cervi (Zeder, 1790), Fischoeder, 1901.

Key To SPECIES

A. ‘Testes in tandem} I
B. Testes diagonal 6
1. Testes lobed eS a i! fen e Si dh Be: a: ae
Testes not lobed ... Bs. val he Es a. Rf ie.
2. ‘Testes with two lobes... i es s. ... P. gigantocotyle
Testes with more than two lobes =f oes i Me a “thea
3. Anterior sucker deeply retracted ey os tnd Pita a ie
Anterior sucker not retracted... a ne oe ane 4
4. Laurer’s canal opens posterior to excretory pore tg ie ¥ P. cervt
Laurer’s canal opens anterior to excretory pore At an P. orthocoelium
5. Laurer’s canal opens posterior to excretory pore me 7 P. liorchis
Laurer’s canal opens anterior to excretory pore we eee =P. wagandt
6. ‘Testes lobed, Laurer’s canal posterior to excretory pore oe P. explanatum
Testes not lobed, Laurer’s canal anterior to excretory pore... P. buxtfrons

* Cohn (1904) created a new sub-family Diplodiscinae, all the members of which are found in
Amphibians and Reptiles. This sub-family has two oral diverticula and has no constant characters
by which it may be distinguished from Cladorchinae.

t Refers to mature worms, because in young specimens of P. cervi, the testes are sometimes found
to be slightly diagonally arranged; by this it is meant that the testes are only slightly out of the
mid-line, so that the antero-posterior axis of the worm always intersects both testes, but one testis
always lies so far in front of the other that a transverse line can be drawn between them without
touching either. In young specimens of P. explanatum, the testes lie one on each side of the mid-line,
so that the antero-posterior axis passes between them without touching either, but one testis is only
slightly in front of the other, so that a transverse line intersects both testes. In mature P. cervi the

slightly diagonal arrangement is not seen, and the testes appear in tandem, whereas in mature
P. explanatum the testes are definitely diagonal.

117
Paramphistomum cervi (Zeder, 1790), Fischoeder, rgor.
SyNONYMY :—

Paramphistomum gracile, Fischoeder, 1901.

Paramphistomum bothriophoron (Braun, 1892), Fischoeder, 1got.
Paramphistomum microbothrium, Fischoeder, 1901.
Paramphistomum bathycotyle, Fischoeder, 1901.
Paramphistomum epiclitum, Vischoeder, 1904.

Paramphistomum papillosum, Stiles and Goldberger, 1910.
Paramphistomum papilligerum, Stiles and Goldberger, 1910,
Paramphistomum indicum, Stiles and Goldberger, 1g1o.

The following is a list of the material available :—

1. ‘len bottles from the stomach of bullocks killed in the military slaughter-
house at Sierra Leone, West Africa.

2. ‘Iwo bottles from the stomach of bullocks killed at Khartoum, Sudan.

One bottle from the stomach of a bullock killed at Yola, Northern
Nigeria.

One bottle from the stomach of a bullock killed at Zomba, Nyasaland.

One bottle from the stomach of a bullock killed at Blantyre, Nyasaland.

One bottle from the stomach of a bullock killed at Accra, Gold Coast.

One bottle from the stomach of a bullock killed at Nairobi, Kenya Colony.

One bottle from the stomach of an antelope (Cobus sp.) shot in the
Northern Territory of the Gold Coast.

9g. One bottle from the stomach of a Roan shot in Nyasaland.

10. One bottle from the stomach of a bullock in England.

11. ‘Two bottles, host and locality ?

Se eae a

Many of the above collections comprised some hundreds of specimens,
and other worms of the same group occurred along with them in several
instances.

EXTERNAL ANATOMY

Size and shape. The size and shape exhibit such infinite ‘variation as

to be of little diagnostic value (Plate V).
’ Cuticular papillae. Fischoeder (1903) states that in some cases he found

a number of small papillae of varying size on the cuticle surrounding the
oral opening of P. cervi, and he does not agree with von Blumberg (1871)
that these papillae are always present. The present investigation is in
accordance with Fischoeder’s view, as they were found in some specimens
and not in others.

Other points usually discussed under External Anatomy will be
included under Internal Anatomy, because their full consideration involves
details that can only be made out in sections or in cleared specimens.

118

INTERNAL ANATOMY

Muscular system. The points requiring special mention in this system
will be referred to under special organs.

Nervous system. This system was not investigated as it has not been
used in specific diagnosis. |

Excretory system. Fischoeder (1903) describes and figures the excretory
bladder of P. cervit as being of a definite shape, with thin smooth walls
and with the excretory pore opening well in front of the bladder.
Examination of a large number of sectioned specimens leads one to the
conclusion that a definite form cannot be assigned to the excretory bladder,
which varies according to its degree of distension. The position of the
excretory pore in relation to the bladder is also found to vary with the age
of the individual ; in those with no eggs in the uterus, the pore opens
dorsally or dorso-posteriorly of the bladder, and as the uterus gradually
fills with eggs the pore appears to pass further and further forward until
it is seen to open well in front of the bladder. This point is fully discussed
under P. explanatum, and as P. cervi is found to vary in exactly the same
manner no further reference to it will be made here.

Anterior sucker. The anterior sucker may be longer than broad,
circular, or even broader than long. When contracted the central canal
of the sucker may be narrow ; on the other hand if its external aperture
is widely open, the canal may be funnel-shaped, becoming narrower towards
its junction with the oesophagus. The internal surface of the oral cavity
may, or may not, be furnished with small papillae, but these vary in
exactly the same way as in the case of the cuticular papillae ; in fact,
owing to their irregular distribution, papillae may be present in some
sections and absent in others of the same worm. Fischoeder (1903) states
that he found papillae in all the sectioned specimens he examined.

Oesophagus. No special mention of the muscle wall of the oesophagus
of P. cervi is made by Fischoeder (1903), but in describing other species,
e.g., P. dicranocoelium and P. cotylophorum, he says that the oesophagus
of these two worms is characteristic, because the muscle wall is thicker
near the gut fork than near its anterior end; therefore the inference is
that the oesophagus of P. cervi does not exhibit this thickening. Careful
examination of five specimens of P. cervi cut in sagittal section gave the
following results.

119

TasBLe I.

Thickness of the muscle wall of the oesophagus of P. cervt.

Near its anterior end Near the gut fork
Specimen 1 ee Ae ae ae se a 324 6ou
Specimen 2 a = = pas Gn eae 324 40
Specimen 3 sas a oe rs: Sas ‘3 20M 4oul
Beeeuriend @y(tt..1\) sel fwd.: yor os HEE «3s 16 36u
Specimen 5 ie ar ti es ey) ie 161 32M

In specimens I and 2, the oesophagus was shorter than in the other
three. From the above it is clear that there is a gradual thickening of the
muscle wall of the oesophagus in P. cervi, but it is not so marked as in the
case of the other two species quoted above, and therefore it is not visible
in whole specimens in P. cervi. Indeed, even in sectioned specimens it is
difficult to appreciate with a low power, because the thickening is very
gradual and not very marked, and it is only when the point is specially
looked for with a high power that one realises its existence. In the
collection of material from the Cobus sp. of antelope in the above list,
some specimens were found which showed the oesophagus to be slightly
bulbous even in whole worms, and on sectioning one of these the muscle
wall of the oesophagus was found to be 20m thick at the anterior and
72u thick at its posterior end. Although the difference in this case is
considerably greater than among the above specimens, the writer does not
consider it sufficiently marked to be taken as a specific difference, and
merely records it as a probable variant of P. cervt.

The intestine. Both the degree of convolution and the precise point
of termination of the gut caeca are regarded by Fischoeder (1903) as
important points in differentiating between species closely allied to P. cervt.
These two points were examined in about 150 specimens cleared in carbolic
and in about 40 sectioned specimens. Fifty specimens were taken from
one bottle, which were chosen on account of their close similarity in
external appearance, as it was thought by this means that variation due to
artificial influences could be almost entirely eliminated. The result of the
examination of these fifty specimens showed that the caeca were nearly
straight in some cases and in others were distinctly convoluted, and

120

between these two extremes there were others with all degrees of convolu-
tion of the gut, which indicates that a variable species is being dealt with.
This view was further supported by noting a different amount of convolu-
tion of the two caeca in the same worm in several instances. The point of
termination of the two caeca also varied considerably ; in some they
ended quite clear of the anterior border of the posterior sucker, and in
others they extended as far as the middle of the posterior sucker. In
collection No. 10 of the above list, the worms were more fully extended
than in any other collection, and in one of these specimens the gut was
found to end 1-6. mm. in front of the anterior border of the posterior
sucker. All gradations between the two extremes were seen, and in a few
worms the caeca ended at different levels on the two sides. The remaining
specimens which were not so uniform in size and shape were found to
vary in the same way. It is also stated that the gut caeca terminate
in a dorsally directed blind end. This was found to be so in most cases,
but three specimens were seen in which the terminal part of the caeca ran
ventrally. It is considered, as a result of these findings, that small
variations in the gut caeca are not reliable points on which to separate
species.

Posterior sucker. The ratio of the diameter of the posterior sucker
to the length of the worm is used in many instances as a distinguishing
feature. For the purpose of examining this character eighty-nine worms
from one bottle were measured ; all of the specimens used were fixed
in a well-extended condition, differences in length from artificial causes
such as shrinking being thus practically eliminated. In these eighty-nine
specimens the ratio of the diameter of the posterior sucker to the length
of worm varied greatly, all intermediate figures between 1:8 and 1: 3°5
being found, and the variations were so gradual that it was found impossible
to draw a dividing line at any point in the series. A large number of
worms from other bottles were also measured in the same way, and as
these were in different degrees of contraction the variation was found to
be even greater than in the above series; in one strongly contracted
example the ratio of sucker to length of worm was only 1: 2:5. The
collection obtained from England (No. 10) consisted of twelve specimens,
which were part of the collection mentioned by Pillers (1922). They were
all fully extended and presented a very uniform appearance externally,
both in size and shape. Five of these specimens were cleared in carbolic
and it was found that the diameter of the posterior sucker in proportion

I2T

to the length of the worm in the five specimens was as follows: I : 9:5,
I:7, 1:57, 1: 5:4, 1: 4:75. All of these specimens were gravid, and
as they only varied from about I1 mm. to Io mm. in length, it is obvious
that the size of the sucker varies quite apart from the age and size of the
worms. According to Pillers these worms are very uncommon in England,
so it is improbable that more than one species is present in this small
collection. All of these worms were cleared in carbolic acid, so that the
general anatomical details were sufficiently clear to render it certain that
only one species was being dealt with, and it was noted that the other
characters used as distinguishing features between species of this group
did not always occur in worms with a corresponding size of sucker. For
instance, a worm with a sucker only one-eighth the length of the worm,
and hence belonging to P. gracile on this account, might have its other
diagnostic points more closely allied to P. cervi or any other of the species
to be discussed below. This mixture of characters of more than one type
could be indefinitely extended until the species became inextricably
confused. It is considered, on account of the above observations, that
the ratio of the posterior sucker to the length of the worm is much too
variable to be used as a diagnostic character.

Genitalia. Testes. The size and shape of these organs and their
relations to one another and to other structures were found to vary some-
what, depending on differences in age and on the degree of contraction in
which the worms happened to be when fixed. One testis always lies
behind the other, in or near the mid-line ; as a rule one testis is directly
behind the other, but in young specimens with small testes they are
sometimes slightly diagonally placed. When the testes have grown,
however, this diagonal arrangement is no longer so obvious. Also in
young worms, and even in fully extended examples with eggs in the uterus,
there is sometimes a distinct interval between the two testes up to as much
asiImm. _ In these instances the testes are circular in outline. On taking
a series of worms of gradually increasing age and degree of contraction, the
testes are found to come closer and closer together until they touch ; after
this they tend to become flattened with consequent extension laterally and
dorso-ventrally, so that they appear oblong in shape and their borders
approach nearer the periphery ; in extreme cases they even cause a bulging
of the external surface of the worm. The relation of the posterior testis
to the posterior sucker also varies considerably ; in young worms and well-
extended adults the hinder testis lies altogether in front of the posterior

I22

sucker, but in worms not so well-extended the posterior border of the
hinder testis reaches, or overlaps, the anterior border of the sucker.
The testes are always diviced into lobes, which are more distinct in young
worms than in older ones. These conclusions are based on the examination
of over forty specimens.

Another point of some interest was that three or four specimens were
found with the uterus quite full of eggs and the testes small and indefinite.
From this it seems possible that the testes atrophy after fulfilling their
functions.

Vas deferens. Fischoeder (1903) made an arbitrary division of this
organ into three principal parts dependent on the anatomical characters
of each part, and as this division is very useful for purposes of description
his nomenclature will be followed in the present paper.

The three portions of the vas deferens are as follows :—

1. Vesicula seminalis. = A thin-walled coiled tube formed by the
junction of the two vasa efferentia.
2. Pars musculosa. = The continuation of the vesicula seminalis,

furnished with a fairly thick muscular wall.

3. Pars prostatica. = A shorter portion of the tube surrounded
by a collection of large cells, the prostatic cells.

The pars prostatica leads into a duct known as the Ductus Ejaculatorius,
which unites with the termination of the uterus within the genital papilla,
and which is known as the Ductus Hermaphroditicus.

The vesicula seminalis varies greatly in size and amount of
convolution in different specimens. These variations depend on
several factors, such as the degree of contraction of the worm, and
whether the vesicula is empty, partly filled, or fully packed with
spermatozoa when the worm is killed. The pars musculosa is liable to vary
from the same causes, and to these must be added variation due to the
state of contraction of its own muscular wall. The muscle wall of the pars
musculosa is composed of two layers, an outer longitudinal layer, and an
inner circular layer, so that contraction or relaxation of one or both of these
layers can cause considerable variation in the length, diameter, amount of
convolution of the duct, or the proportionate thickness of the two muscle
layers. Both the vesicula seminalis and the pars musculosa coil freely on
themselves and on each other, so that their relations with each other vary
greatly in different specimens, and even in different sections of the same
specimen. From examination of a large number of sectioned specimens,

123

the writer has come to the conclusion that the relations of the vesicula
seminalis and the pars musculosa are so variable that it is not possible to
assign any special type of relation between these two parts to any one
species aS Fischoeder does in several instances. The pars prostatica
_ varies considerably in size and shape ; this fact is clearly brought out in
Table II, which is compiled from the measurement of fifteen individuals
cut in sagittal section and mounted serially ; the maximum measurements

are given in every case.
Taste II.

The length and breadth in microns of the pars prostatica of fifteen specimens of P. cervi.

Length Breadth Length | Breadth

I 178 178 9 436 | 257.
2 218 138 10 475 | 297
3 218 218 II 535 ) 396
~ 257 198 12 594 | 317
5 277 198 13 594 396
6 297 178 14 594 475
7 317 138 15 594 | 495
8 | 317 218

This table shows a regular gradation of sizes from the smallest to the
largest, and from this it is concluded that it is not possible to separate
species on this character.

Genital pore. The genital pore usually lies about opposite the gut
fork or a little behind it, the variation in position depending on the
length and course of the oesophagus, which naturally has a direct effect
on the position of the gut fork. But in one sectioned specimen which
was fully grown and very much contracted, and in consequence only
about 4 mm. in length, the genital pore was only 1/4:8 of the body length
from the anterior end of the worm, and it lay opposite the junction of the
middle and posterior thirds of the anterior sucker, that is, far in front of
the gut fork. This is not in agreement with the statement of Fischoeder
(1903) who, in his description of P. cervi, says:

‘Die Geschlechtsiffnung liegt am hinten Ende des vordern Kérperdrittels in

der Hche oder kurz hinter aber niemals vor der Gabelstelle der Darmschenkel.’

124

Genital atrium and genital papilla. These two structures are used
extensively in specific diagnosis both by Fischoeder (1903) and by Stiles and |
Goldberger (1910). The papilla is composed almost wholly of muscle,
and the atrium is surrounded bya special thickening of the subcuticular
layer. Accordingly, both are liable to great alterations in form, and
further, the papilla is capable of being withdrawn deeply into the body of
the worm with resultant deepening of the genital atrium, or else it can be
completely extruded through the genital pore, in which case the genital
atrium disappears. As would be expected, many worms show intermediate
stages between these two extremes. The variations of the genital apparatus
are fully discussed in P. explanatum (see fig. 2), and as exactly the same
type of variations are found in P. cervi the reader is referred to
P. explanatum. The conclusions to be drawn from these facts are that the
presence of one or two chambers in the genital atrium, or even the total
absence of this cavity, or the size and shape of the genital papilla, are
purely a matter of chance and are of no use in distinguishing various
species. If Fischoeder’s figures (1903) of P. cervi are referred to, it will
be noted that in fig. 1 the pore is shown as a small cavity with no sign of
the papilla protruding, whereas figs. 2 and 3 show the papilla protruded
through the pore and there is no sign of an atrium at all. This suggests
that Fischoeder recognised the possibility of variation in these structures
without referring to it in the text. Small papillae are also described as being
found on the internal surface of the genital atrium in some species and not
in others. Examination of a number of sectioned specimens has shown
that these papillae may be present or absent when the atrium is present,
and a further fact which makes these structures still more liable to variation
is that often the atrium itself is not present.

Ovary. It is recognised by all observers that the ovary may vary
considerably in position in the same species. The present investigation
fully bears this. out.

Shell gland. This gland always lies close to the ovary, and so varies
in company with it.

Vitellaria. The vitellaria appear to be the most variable of all the
organs in P. cervi, and the number of gland groups, their size, and their
distribution, undoubtedly increase considerably as age advances. But
even when comparing them in over one hundred specimens in which the
uterus was well filled with eggs they were found to show marked variation.
Among these hundred odd specimens the anterior limits of the vitelline

125

glands were found to lie as far forward as the hinder end of the anterior
sucker in some cases, whereas in others they did not reach as far as the
genital pore, and between these two extremes all degrees were found.
Sometimes in the same worm the glands did not reach the same level on
both sides. The posterior limits of the vitellaria were also found to vary
considerably, but as a general rule they ended a little behind the termina-
tion of the gut caeca; but this was not invariably the case, for in a few
instances the vitelline follicle groups were found extending to the extreme
posterior end of the worm and could be made out in the parenchyma
surrounding the opening of the posterior sucker. The degree of extension
of the vitellaria inwards on the dorsal and ventral surfaces also varied
markedly. In some cases the glands were strictly limited to the area
outside the intestines on each side, and from this limited distribution all
stages of inward extension were found up to a point where the glands of
the two sides were practically continuous with one another all along both
surfaces. It was noted, however, that when the glands were widely
spread they seemed to be sparsely distributed, and when fairly circum-
scribed in their distribution the groups of follicles were much more closely
gathered together. Fischoeder (1903) uses slight differences in distribution
of the vitelline glands for specific diagnosis ; the writer does not consider
this possible, because in his series of over one hundred gravid specimens of
P. cervi a much greater range of variation was found than Fischoeder
describes in his different species, and between the extreme limits of these
variations all possible degrees were found which made it impossible to
separate one species from another.

Uterus. The dorsal antero-posteriorly directed portion of the uterus
is described as being more convoluted in some species than in others.
After a large number of specimens were examined it was realised that the
amount of convolution the uterus shows is subject to a wide range of
variation in P. cervi. This variation was found to be dependent on two
factors, first, the amount of contraction of the worm, and second, the
number of eggs in the uterus. It was found that in specimens with no
eggs the uterus was nearly straight, but as it became more and more filled
with eggs the degree of convolution of the uterus increased also. It is
therefore considered that slight differences in the amount of convolution
of the uterus cannot be taken into consideration in specific diagnosis.

Eggs. Eggs taken from the uterus of several preserved specimens of
P. cervi were found to vary considerably in size, being from 114 in length

126

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127

by 60u in breadth up to 176 in length by gon in breadth. The above
figures represent the extremes of size found after several specimens had been
dissected ; the maximum variation in eggs from a single worm was only
about 10m in length and 7y in breadth. It is considered on this account
that small differences in size of the eggs are not reliable characters for
specific diagnosis, at all events when the eggs are taken from the uterus of
preserved specimens. It is possible they may be more uniform in size
after being laid.

Laurer’s canal. ‘The relations between Laurer’s canal, the excretory
bladder, and excretory pore are used by Fischoeder (1903) in distinguishing
certain species from one another. He evidently attaches great importance
to these relations, because he divides the genus Paramphistomum into
three groups on these characters, and Stiles and Goldberger (1910) have
followed Fischoeder in this respect. Fischoeder’s three groups of the
genus are as follows :—

1. Laurer’s canal crosses the excretory bladder completely. This
means that Laurer’s canal opens behind the excretory pore and in the
mid-line of the worm.

2. Laurer’s canal and the excretory bladder do not cross. This means
that Laurer’s canal opens in front of the excretory pore.

3. Laurer’s canal crosses the excretory bladder incompletely. This
means that Laurer’s canal opens behind the excretory pore, but to the
side of the mid-line, i.e., the same side as that on which the shell gland lies.

On account of the importance attached to these characters, five
specimens of P. cervi all from one bottle and as nearly as possible similar
in external appearance, were cut in transverse sections, 25m thick, and
mounted serially, so as to test the value of the above statements. The
result of this investigation is shown in Table III.

Table III and Fig. 1, which consists of camera lucida drawings of
Specimens A, B, and C in Table III, show that the relations of Laurer’s
canal to the excretory bladder and excretory pore vary considerably, and
in some cases the relations of these structures do not come under any of
Fischoeder’s three headings. It is probable that if more worms were
examined still other arrangements would be found, but it was considered
that the above five specimens sufficiently proved that these relations were
unreliable for diagnostic purposes. Although only the above five specimens
are included in the table, about thirty others were examined; these
were cut in sagittal and coronal sections and the above results were thereby

Length

(1) P. cervt

5-12 mm.

Shape

Posterior end rounded

Relation of
sucker to
length of

worm

T 34 COgtesaS

Genital pore

length from anterior end.

Opposite or behind gut fork, $ of body

128

(z) P. gracile

11-15 mm.

Almost cylindrical.

Well behind gut fork, } of body length

from anterior end.

Vesicula
seminalis

Pars
musculosa

Pars
prostatica

Papillae on

anterior
end and in
* pharynx’

Termination
of gut

caeca

Testes

Very broad, thin walled, in front of

anterior testis.

o°8 to ro mm. long, walls 18/0 to 224

thick, varying with its degree of
fulness. Straight or slightly coiled.

3004 to 6004 in transverse diameter.

On external surface and in anterior
portion of ‘ pharynx.’

Dorsal to sucker.

dorso-
trans-

2 mm. to 2°8 mm.

I°s mm. to 2 mm.
verse.

PosTERIOR: 2°8 mm. to 3°5 mm. dorso-
ventral. 1 mm. to I°5 mm. trans-

verse.

Slightly on opposite sides of middle

line, close to ventral surface.

ANTERIOR:
ventral.

Vitellaria

ANTERIOR: Oval, 1°2 mm. by 0°7 mm.

Round canal in median plane.

5004 to booyt long, walls 184 to 2240

thick. Moderately straight.

50074 to boot long by 250f4 to 350"
broad.

On external surface and not in

* pharynx.’

Anterior to sucker.

slightly dorsal.
Posrertor: More rounded, o'9 mm.
by 1'o mm.,_ slightly
ventral.
Only lie slightly on opposite sides of

median line.

Extend from ‘ Pharynx’ to posterior
border of ventral sucker. Extend on
dorsal and ventral surfaces. In coarse
groups of follicles close together.

Ovary

Uterus

Eggs

Laurer’s
canal

Excretory

bladder

* At the first glance P. botbriophoron appears to have several rather marked points of difference from the other five species, €2
pars musculosa is closely coiled; the pars prostatica is very long; and the genital atrium is large and deep, with no genital pap
musculosa.

Runs dorsally towards anterior and

right or left of mid-line.

Portion dorsal of testes markedly wavy.

145-156 X 75-82.

opens in mid-line about level of
posterior border of the hinder testis.
1-1'2 mm. behind excretory pore.

Flask-shaped, close to dorsal surface.
Pore opens anteriorly in mid-line.
Crossed by Laurer’s canal at junction
of anterior and middle third.

Close behind base of sucker, either to

115-125ft X 72-804.

Extend from hinder’ border of
‘Pharynx’ to anterior border of
sucker ; do not extend markedly on
dorsal and ventral surfaces. In five
groups of follicles somewhat irregu-

larly placed.

Behind posterior testis either to left or
right of mid-line and slightly towards
ventral surface (i.e., anterior to
sucker).

Portion dorsal of testes wavy.

Curves posteriorly and opens at level
of anterior border of ovary 1°5 mm.
behind excretory pore. :

Further from dorsal surface. Pore on a
level with hinder border of posterior
testis. Is crossed by Laurer’s canal
about its centre.

Summarising the chief characters described by Fischoeder (1903

(3) P. microbothrium

Somewhat more flattened dors
ventrally than P. cervi. Only slight
curved ventrally like P. gracile, .

¢§

Ls Attogl 3463 |

Behind gut fork, $ of body length fro
anterior end. Has a genital atriu
and more definite muscular sphinct,

Lies dorsal of pars musculosa, unli

P. cervi and P. gracile where it’
behind. 2 |

Walls 45s0-soys thick: almost excl
sively consisting of circular muse
with a single outer layer of longit
dinal muscle. Strongly coiled. |

5004 to 6oop long.

me aac

me!

Not mentioned.

diene gr ei

ele Fe See Manat Me

Anterior to sucker.

ANTERIOR: 2°3 mm. to 2°5 mm. dort

ventral.

1°5 mm. to 1'7 mm. tral’

verse and longitudina.

Posterior: As a rule somewhat larg

A little more markedly on oppos

sides of mid-line. Close to |
:

surface. |

. . z )
On one side reach from posterior bore
of ‘ Pharynx’ to anterior border |
sucker. On other side from gut fc
to middle of sucker. Similar ;
P. cervi in size and extent on dor
and ventral surfaces.

Further from mid-line than in P. cei

and P. gracile. ;

As in P. cervt. } .
145-150ft X 75-80ft. j

Opens to one side and not in mid-lin
opposite anterior border of ova,

250-30044 behind excretory a. |

Similar to P. cervi. Pore in mid-lt
Laurer’s canal does not cross bladd

———————

(4) P. epiclitum

-|5-9 mm.

~ 1904), as typical of the following six species of Paramphistomum.

129

(5) P. bathycotyle

(6) P. bothriophoron*™

11-15 mm.

Anterior } strongly curved ventrally, the
remainder straight, as in P. cervi: the
greatest circumference at 2nd and 3rd

thirds.

Ventrally curved. Greatest transverse
diameter near posterior end.

tes tO: 4°5.-

At junction of 1st and middle thirds. Well
behind gut fork.

Dorsal and posterior of pars musculosa.
| .

Walls 18 to 22 thick.

6oout-Soou, and 250-3004 thick.

Unlike P. cervi, which is round, it is long | Almost globular,

In text 2? of body length; in drawing 4 of
body length.

About middle of anterior third behind gut

fork. Very little muscle surrounds it.

Lies loosely coiled between intestines.

6-9 mm.

Of 4 specimens: 2 strongly curved ;
slightly curved ventrally.

Not mentioned.

In middle of anterior half. Not so well
developed as in other species. Like
P. microbothrium.

Behind and not dorsal to pars musculosa.
Closely coiled.

Ventral and anterior to vesicula, 0°6-0°75

mm. long and 184-22 thick, not coiled.

4ooul-5004 in diameter.

Strongly developed, not loosely coiled like
others, but closely coiled. Muscle
different from all other species because,
unlike those which have nearly all circular
muscle surrounding a single layer of
longitudinal, the longitudinal is nearly
as thick as the circular.

1'0-1'2 mm. long.

Not mentioned.

a. base of sucker.

1°8-2°2 mm. dorso-ventral.
o*g-1°2 mm. longitudinal.
1°2-1°6 mm. transverse.

2°1-2°5 mm. dorso-ventral.
or7-o0'1 (? 1°0) mm. longitudinal
1°6-2°O mm. transverse.

PosTeRIor :

Of different sizes and very irregular, but
close together. Stretch from beginning
of oesophagus as far as base of sucker.

| Reach further on dorsal and ventral

' surfaces than in P. cervi.

Between posterior testis and base of sucker
and on same side of mid-line as anterior
testis.

As in P. cervt.

——

}

145-155u x 75-80u.

Opens in mid-line about the level of its
_ ‘Origin.

Opens on level of middle of posterior testis
500-6004 anterior to Laurer’s canal.
Laurer’s canal crosses between 1st and

2nd thirds of bladder.

II$-125u X 70-75p.

On both external surface and in ‘ pharynx.’

Not seen.

Close in front of anterior border of sucker.

Longitudinal : 1°0-1°3 mm. \ Both about

Dorso-ventral : 1°5-1°8 mm. { same.
Like P. cervi, shghtly out of mid-line.

Almost from ‘ pharynx’ to opposite anterior
phary pp

border of sucker. Almost confined to
lateral fields only slightly encroaching on
dorsal and ventral surfaces. Single follicles
small.

Similar position to P. cervi, but only

slightly away from mid-line.

Broad and filled with eggs.

Runs directly dorsal and opens on a level
with ovary.

Different from P. cervi because it has a
long anteriorly directed canal which opens
opposite posterior border of anterior testis
about middle of length of worm.

Close in front of sucker.

ANTERIOR: 0°7-0°8 mm. long.

PosTertor : 0°8-1'o mm. long.

Both 2°0-2°3 mm. dorso-ventral. More
deeply lobed. Lie in similar position to
P. microbothrium.

From gut fork almost to middle of sucker
and also extend on dorsal and ventral
surfaces.

Between hinder testis and sucker very near
ventral surface and very much to the side.

Filled with eggs.

125-1354 X 65-70.

Opens opposite side to ovary as in P. micro-
bothrium, opposite anterior border of
shell gland.

More rounded; in front of sucker near
dorsal surface.

_ the longitudinal muscle of the pars musculosa is said to be very thick, and it is figured as being arranged in distinct columns ; the
It is conceivable that all of these conditions could be caused by one factor, viz., contraction of the longitudinal muscle of the pars

130

confirmed. But one point was found to be of value in regard to the
relations of the opening of Laurer’s canal and the excretory pore, and
that is, that in species in which Laurer’s canal is described as opening
behind the excretory pore this is invariably the case, although the actual
distance at which one pore lies behind the other varies a good deal.

BI.

Fic. 1. Paramphistomum cervi, transverse sections. A1 and Az—Sections of one specimen
at different levels. 81 and B2—Sections of a second specimen at different levels. C—Section of
a third specimen passing through the excretory pore. ex.b.—excretory bladder ; ex.d.—excretory
duct; ex.p.—excretory pore; i.—intestine; Z.c.—Laurer’s canal; ov.—ovary; sh.g.—shell
gland; ut.—uterus; v.g.—vitelline gland. x 16.

Careful examination of the descriptions of the six species of
Paramphistomum in Table IV shows that the differences between them
are very minute, and a further comparison of this table with the results of
the extensive examination that has been carried out by the writer shows that
with one exception all the characters used by Fischoeder as of specific

131

value in distinguishing these six species come within the range of what
appear to be variations of P. cervi. The exception is the length of the
pars prostatica; the maximum length observed by the writer for this
organ was 594m, whereas Fischoeder gives it as 800m in P. epiclitum,
and 1-2 mm. in P. bothriophoron. But this is such a small difference that
the author is inclined to disregard it and to consider it probable that the
above species are all one.

There are three species, P. papilligerum, P. papillosum and P. indicum,
described by Stiles and Goldberger (1910) which also appear to the writer
to be identical with P. cervt.

Paramphistomum papilligerum, Stiles and Goldberger, 1gro.

Apparently the material on which this species was made consisted of
a series of frontal sections loaned to the authors by Shipley.

The only difference between this species and P. cervi is that
P. papilligerum is described as having small papillae on the inner surface
of the genital atrium. It has already been shown that papillae in this
position may be present or absent in P. cervi, and further that the presence
of a genital atrium itself is a variable character. There is hence no
justification for the separation of this species.

— Paramphistomum papillosum, Stiles and Goldberger, 1910.

The material at the disposal of the authors is stated to be a single
non-gravid specimen. The species is. characterised by the presence of
papillae on the anterior end of the worm, in the oral cavity, and in the
genital atrium. Papillae in these positions have already been shown to
be variable characters of P. cerv1, being present or absent in one or all of
these positions. In P. papillosum the excretory pore is stated to be dorsal
to the excretory bladder, and in P. papilligerum it is stated to be anterior
to the bladder ; but it must be remembered that in their descriptions of
these two worms, Stiles and Goldberger state that eggs were present in the
uterus of P. papilligerum, and not in P. papillosum. It has already been

shown that in P. cervi the excretory pore appears to pass forward as age
advances ; this fact is in agreement with the difference observed between
the species P. papilligerum and P. papillosum, when it is remembered
that the former is probably a younger specimen than the latter. It is
therefore considered that P. papillosum is synonymous with P. cervi.

tebe
Paramphistomum indicum, Stiles and Goldberger, 1910.

These worms were said to have been found in two bottles along with
other species, and no eggs were observed. The description and drawings
of this species by Stiles and Goldberger agree in all points with many
specimens of young P. cervi without eggs in the uterus, examined by the
writer during the course of the present investigation. That the writer’s
specimens referred to were young P. cervt and not another species is
rendered practically certain, because in the same bottles worms in all stages
of development could be found, so that the gradual change from worms
similar in appearance to P. indicum could be followed in a long series
until a typical P. cervi with uterus full of eggs could be found. The writer
is therefore of the opinion that on its present description P. imdicum
cannot be distinguished from young P. cervt.

In the writer’s opinion P. gracile, P. bothriophoron, P. microbothrium,
P. epichitum, P. bathycotyle, P. papilligerum, P. papillosum and P. indicum
are all synonyms of P. cervt.

Paramphistomum lorchis, Fischoeder, Igor.

This worm is easily distinguished from other species of the genus,
because the testes are not lobed ; it is the only species in which the testes
exhibit this character, and in which Laurer’s canal opens behind the
excretory pore.

Paramphistomum pisum, Leiper, Igro.

The material available for examination consisted of two collections,
about thirty specimens in all. — The location of the parasite in the host
(Hippopotamus) is not given, but Leiper (1910) states that it is found in
the small intestine.

No gravid specimens were observed by the writer, but about half the
material was sexually mature. Leiper says the worms are pisiform when
fresh and somewhat contracted when preserved. This agrees with the
writer's observations for all of his specimens were slightly contracted with
the result that they were almost globular in shape, the largest measuring
about 3mm. in diameter. The worms agreed essentially with Leiper’s
description ; both suckers communicate with the exterior by narrow
canals as Leiper figures them, but in three specimens which the writer

133

sectioned the anterior sucker was found immediately beneath the cuticle
as in other members of the genus, and is not separated from it by a
canal of considerable length as in Leiper’s fig. 34.

Paramphistomum gigantocotyle, Brandes, 1896.

Host :—Hippopotamus amphibius. Location :—Stomach. Locality :—
Africa.

As the original description of this species is somewhat inadequate,
Leiper (1910) redescribed it.

According to Leiper this worm may be distinguished from other
species by the relative size of the posterior sucker and by the shape of the
testes. Although the diameter of the posterior sucker given by Leiper,
viz., 32 mm. in a worm of 8 mm. in length, is relatively great, it is not
by itself a sufficient character for identification, because in P. explanatum
the relative size of the sucker is often more than this. But the testes
appear to be characteristic, as although they are placed one behind the
other as in P. cervt, they differ in that whilst in the latter they are divided
into several lobes, in P. gigantocotyle each testis is nearly completely divided
into two portions by a deep transverse groove, so that in sections they may
appear as four organs, a condition that is never found in P. cervt.

Paramphistomum explanatum (Creplin, 1847), Fischoeder, r1gor.

SYNONYMY :—

Paramphistomum calicophorum, Fischoeder, 1901.
Paramphistomum crassum, Stiles and Goldberger, 1910.
Paramphistomum cauliorchis, Stiles and Goldberger, 1g10°
Paramphistomum fraternum, Stiles and Goldberger, 1910.
Paramphistomum siamense, Stiles and Goldberger, 1910.

The material available consisted of the following collections :—

I. One bottle from the stomach of a bullock killed at Durban, South Africa.

2. One bottle from the stomach of a bullock killed at Townsville, Australia.
(Over 100 specimens.)

One bottle from the stomach of a bullock killed at Khartoum, Sudan.

One bottle from the stomach of a bullock killed at Blantyre, Nyasaland.

Two bottles from the stomachs of two hartebeests shot near Blantyre,
Nyasaland. (Both bottles contained over 100 specimens.)

ite ware:

This worm is readily distinguished from P. cervi, because the testes are
always diagonally situated one overlapping the other, both laterally and
antero-posteriorly, in fully grown worms, whereas in full-grown P. cervt,
the testes lie one directly behind the other.

134

At the beginning of his decription of P. explanatum, Fischoeder (1904):
states that this species most nearly resembles P. bathycotyle on account of
its shape, and throughout his paper he contrasts these two species without
reference to others. Plate VI, fig. A, clearly indicates that shape cannot
be taken as a diagnostic point ; the fact that the testes in P. bathycotyle are
placed antero-posteriorly at once distinguishes it from P. explanatum and
renders further comparison of these species unnecessary. Apart from the
arrangement of the testes the general anatomy of P. explanatum is very
similar to that of P. cervt and the organs are found to be subject to similar
variations. The relatively large size of the posterior sucker is apparently
regarded as an important point in distinguishing between P. explanatum
and other species. Fischoeder (1904) states that the worm varies from
8mm. to 13 mm.in length. But in giving the size of the posterior sucker,
he makes use of a single specimen 8 mm. in length and gives no particulars
of the dimensions of the suckers in larger worms. The sucker of this
single specimen is stated to be 3-5 mm. in its antero-posterior diameter,
and 3-0 mm. transversely. That is, the greatest diameter of the sucker
to the length of the worm is as I: 2:3.

Ten specimens of this worm were cleared in carbolic acid and the
length of the worms and the size of the posterior sucker ascertained ; these

are given in Table V.
TaBLe V.

Measurements of ten specimens of P. explanatum.

Ratio of diameter of

Specimen Length of worm Diameter of sucker sucker to length

in mm. in mm. of worm
I 10°] 35x24 12 RG
2 9°7 2:SPa8 12 3°6
3 9°6 2°9 X 2°7 LBD i:
4 8°5 ie Rot eee us P24
5 7°6 oe te ee Tr tee
6 6°8 B15 Pais fe ES
7 6°8 SEX 315 Lie 39
8 6°6 26 4 2°6 148 2u5
9 6°6 2:0 eo ips
fe) 6°0 2'64% 2°6 L22"3

Average I : 2°5

135

From this table it is clear that the ratio of the diameter of the sucker
to the length of the worm varies above and below Fischoeder’s figure, which
closely approximates to the mean. On the whole the sucker is relatively
larger than in P. cervi, but it is subject to such variations that it cannot
be taken as an absolute guide in diagnosis. Another point brought out in
the above table is that the sucker is not typically oval in shape, being only
occasionally met with in this form.

In describing the genital apparatus, Fischoeder states :

‘Auch das genitalatrium ist nur sehr klein. Die dasselbe umgebende, von dem
ubrigen Korperparenchym wenig abgegrenzte Musculatum ist nur 0:08-o-l mm.

stark, und die in Grunde des Atriums befindliche Papille ist ebenfalls nur ausserst
schwach entwickelt.’

Fig. 2 consists of drawings of the genital apparatus of three specimens
of P. explanatum cut in sagittal section and from these it is clear that
such a precise description as the above of this portion of the worm is not
permissible. Although this point has not been figured in detail in dealing
with other species, the same range of variation in the genital papilla and
genital atrium has been found. Fischoeder says that the genital organs
are displaced towards the anterior end of the worm, and that the testes
reach near to the ventral surface, in which case they are of necessity in
front of the posterior sucker. In some instances this is correct, as is
illustrated in fig. 3, but that the testes are not invariably in this position is
shown in figs. 4 and 5. In these two worms the testes are seen drawn
away from the ventral surface and they he dorsal to the posterior sucker.
In all probability this is an artificial condition brought about by contraction
of the worms at the time of fixation, because both these specimens were
noted to be considerably contracted before they were cut, and this is
borne out by the irregularity of outline shown in the drawings on the
dorso-posterior portion of the worm, especially in the case of fig. 4. This
probability is further supported by the appearance in fig. 6, which is
a sagittal section of an immature well-extended specimen ; in this drawing
the single testis figured is seen to be situated nearer to the ventral than to
the dorsal surface and well in front of the sucker. The four worms from
which the above drawings were made all came from the same bottle, and
specimens in varying degrees of contraction and of different ages are found
with characters intermediate between them.

If fig. 6 is examined, it will be readily understood how by contraction
of the worm the testes are brought to the position seen in the two previous

136

figures. The base of the posterior sucker slopes backwards and towards the
dorsal surface, and antero-posterior contraction of the worm would cause
the testes to impinge on this surface, when they would follow the path of

WO AS
yes uvs . IZA =
S Ta .
FF oS
= io a

Fic. 2. Paramphistomum explanatum. Sagittal section through the genital pore of three
specimens. 4—Genital papilla fully retracted. B—Genital papilla partly retracted.
C—Genital papilla fully extruded. a.s.—anterior sucker ;_d.e.—ductus ejaculatorius; d.b.—ductus
hermaphroditicus ; _g.a.—genital atrium ; g.p.—genital pore 3 g.pp.—genital papilla ;

p-—parenchyma ;_ p.m.—pars musculosa; p.p.—pars prostatica; sc.m.—subcuticular muscle ;
s.m.—sphincter muscle 3 ut.—uterus. X 30.

least resistance and pass up towards the dorsal surface of the worm ; at the
same time the testes exert some pressure on the sucker, causing its base to

Fic. 3. Paramphistomum explanatum. Coronal section of gravid worm. a.s.—anterior
sucker; 7.—intestine ; p.m.—pars musculosa; p.p.—pars prostatica; p.s.—posterior sucker ;
sh.g.—shell gland ; ¢.—testis; ut.—uterus; v.g.—vitelline gland. x 12.

Fic. 4. Paramphistomum explanatum. Sagittal section of gravid worm near the mid-line.
a.s.—anterior sucker ; ex.b.—excretory bladder; ex.c.—excretory canal; ex.p.—excretory pore ;
£-p.—genital pore; i.—intestine ; L.c.—Laurer’s canal; 0.—oesophagus ; ov.—ovary ; p.m.—pars
musculosa ; p.p.—pars prostatica ; .s.—posterior sucker ; sb.g.—shell gland ;_ ¢1.—anterior testis ;
i2.—posterior testis; ut.—uterus; v.g.—vitelline gland ; v.s.—vesicula seminalis. +

Fic. 5. Paramphistomum explanatum. Sagittal section of a partly gravid worm near the
mid-line. a@.s.—anterior sucker ; ex.b.—excretory bladder ; ex.p.—excretory pore; g.p.—genital
pore; L.c.—Laurer’s canal; 0.—oesophagus; p.m.—pars musculosa; p.p.—pars prostatica ;
p.s.—posterior sucker; i1.—anterior testis; /2.—posterior testis; ut.—uterus; v.s.—vesicula

seminalis. x 16.

Sagittal section of posterior part of immature worm

Fic. 6. Paramphistomum explanatum.
L.c.—Laurer’s canal ;

near the mid-line. ex.b.—excretory bladder; ex.p.—excretory pore ;
p.s.—posterior sucker ; t2.—posterior testis; ut.—uterus. x 16.

159

become flatter and also making it look almost directly ventrally instead of
ventro-posteriorly. The full effect of such contraction is shown in fig. 4.

Fischoeder describes and figures the excretory bladder as being of a long
flask-shaped type with a narrow canal running forward from it and opening
about the level of the posterior border of the hinder testis. Figs. 4, 5 and 6
clearly show that the shape of the excretory bladder is very variable and
that the excretory canal runs dorsally from the bladder in young worms,
whereas in older specimens it becomes longer and narrower until, in fully
gravid worms, it opens well in front of the bladder.

Stiles and Goldberger (1910), in their diagnostic key of the genus,
place P. explanatum in the group of worms with the ‘ excretory pore
prevesicular,’ and in the sub-group in which the following characters are
given as diagnostic. ‘ Testicular fields median, coincide or overlap, zones
lobate, testes much smaller than and near the acetabulum; ventral chamber
absent ; excretory vesicle long and narrow ; acetabulum very large.’
Their specific definition is: ‘Genital pore in postbifurcal zone ;
musculosa (?) ; prostatica longer than musculosa ; acetabulum less than half
as long as body ; caeca sinuous, moderately broad, end in acetabular zone ;
body 8 to 13 mm. long ; type host Bos indicus at, Berlin, Germany.’

With the exception of the relation of the testes to each other all of the
above diagnostic characters are so variable that none of them are reliable.

Paramphistomum calicophorum, Fischoeder, Igor.

At the outset of his description of this worm Fischoeder (1903) states :

‘Das Material ist stark geschrumpft, briichig ; trotz Behandlung mit lod und
Wochen langer Einwirkung von Kreosot lisst sich eine zur Untersuchung von ‘Total-
praparaten nur wenig brauchbare Durchsichtigkeit erzielen, mit Ausnahme von
einzelnen unreifen Exemplaren aus dem letzt genannten Glase. Dagegen ist es mir
gelungen, von reifen Exemplaren aus dem Glase No. F’. 659 fiir meine Zwecke einiger
Maassen brauchbare Schnittserien anzufertigen.’

From this it is obvious that he was unable to obtain a dorso-ventral
view of a gravid worm, and, from his figures, he was apparently content
with sagittal and transverse sections of full-grown specimens ; the only
dorso-ventral view he gives is that of an immature worm with the testes
small and wide apart, an arrangement which is quite unlike that found in
gravid specimens. Thus he failed to realise the identical arrangement of
the testes in mature P. explanatum and P. calicophorum.

As the sucker is said to be somewhat smaller in relation to the length

140

of the worm in P. calicophorum than it is in P. explanatum, Table VI has
been drawn up by utilising the figures given by Fischoeder for these two
worms and by summarising the figures in Table V of the present paper.

Tasre VI.
P. explanatum.
P. explanatum, P. calicophorum Present material
Fischoeder summarised from
Table V
Length of worm ... ee oe ...| 8mm. to 13mm. | 1omm. to 15 mm. | 6mm. to Io‘l mm,
No. of worms measured... whe wei p ? fe)
Diameter of posterior sucker... .».| 3°4 mh. X37 mm. 3mm. to 4°3mm. | 2.6mm. to 3.8 mm.
No. of suckers measured 4. isd I ? Io
Ratio of diameter of sucker to Iength : ae F 3 t ,
of worm expressed in fractions th gs We nk ?
2°3 2g 36

It seems clear from Table VI that there is no constant relation between
the diameter of the posterior sucker and the length of the worm.

Fischoeder recognises the variation to which the genital apparatus is
liable and to which attention has been drawn in P. explanatum in the
present paper. He states:

‘Die Hoden liegen fast neben einander (fig. 29, 30), der vordere etwa in der
Mitte des Korpers, mehr dorsal, der hintere mehr ventral (fig. 29, 30 u. 31).’

His references to the figures prove that he is describing the appearance
in young worms, which is borne out by his later statement :

‘ Bei reifen Individuen nehmen sie die ganze, stark verdickte, hintere Hilfte des
Thieres bis dicht zum Saugnapfe ein (fig. 31).’

He makes no reference to the appearance of the testes in immature
P. explanatum.

With regard to the excretory bladder, excretory pore, and Laurer’s
canal, he says:

‘In Bezug auf das Lageverhiltniss des Excretionporus und der Ausmiindungs-
stelle des Laurer’schen Canals sind haufig gewisse Differenzen zwischen den reifen

141

und unreifen Individuen zu verzeichnen, die jedoch auch mit der Entwicklung der
Hoden und der damit verbundenen starkern Ausdehnung der hintern Kérperpartie
in Verbindung gebracht werden kénnen. Wahrend nimlich bei den unreifen
Thieren in der Regel die Excretionsblase etwa im Niveau des vordern Randes des
Saugnapfes und der Laurer’sche Canal nur 0-3-0-4 mm, dahinter ausmiindet (fig. 30,
32 u. Textfig. E), befindet sich der Excretionsporus bei den geschlechtsreifen
-'Thieren meist viel weiter nach vorn, fast in der Héhe des vordern Randes des hintern
Hodens (fig. 31), und die Ausmiindung des Laurer’schen Canals 0-6-0:8 mm. hinter
dem Excretionsporus.’

It is therefore clear that in the case of P. calicophorum, Fischoeder
recognised the variation to which these structures are liable, but, as he
does not seem to have made a detailed examination of immature material
of other species, he failed to realise the general application of this fact.
While in agreement with Fischoeder in the first part of his statement,
viz., that the excretory pore and excretory canal vary in their relations to
the bladder as age advances, the writer cannot support the latter part of
his statement that Laurer’s canal opens further behind the excretory pore
in gravid worms than it does in immature examples. While agreeing
that a variation in this particular can be found, it has been observed to be
quite independent of age.

Stiles and Goldberger (1910), apparently basing their definition of
P. calicophorum on Fischoeder’s paper (1903), write :

‘Excretory pore prevesicular.

Testicular fields separate, not median, zones overlap, testes lobate, much smaller
than acetabulum; . . . excretory vesicle not narrow but swollen iN
From this it is clear they have ignored Fischoeder’s statements quoted
above and have taken their definition mainly from characters found in
immature worms, which makes it useless for diagnostic purposes. Careful
comparison of Fischoeder’s descriptions of the species P. explanatum and
P. calicophorum thus indicates that there are no differences between them,
except such as may be explained by differences in age and normal variation ;
therefore P. calicophorum is a synonym of the species P. explanatum.

Paramphistomum crassum, Stiles and Goldberger, 1910.

Stiles and Goldberger (1910) made the species P. crassum from three
specimens which they state were found with other forms in a bottle from
India, the host being Bos indicus. They also say that no eggs were seen,
so it is probable their material was immature.

On comparing Stiles and Goldberger’s description of P. crassum with

142

Fischoeder’s description of P. calicophorum, the only essential difference
between the two is that the excretory pore is dorsal to the excretory bladder
in P. crassum, and although Stiles and Goldberger assert that the excretory
pore is anterior to the excretory bladder in P. calicophorum, it is not in
agreement with Fischoeder’s statement, quoted above, except in fully grown.
worms, and with this the writer’s observations are in agreement. It has
also been shown that in P. explanatum without eggs the excretory pore is
in the position ascribed to it in P. crassum by Stiles and Goldberger.
When it is recalled that the latter observers saw no eggs it seems probable
that they were dealing with young specimens of the same species as
Fischoeder. ,

Paramphistomum cauliorchis, Stiles and Goldberger, Igto.

In describing this species Stiles and Goldberger (1910) state that
they found three specimens in one bottle and four in another. The host
was Bos indicus and eggs were not observed. In their description of
P. crassum Stiles and Goldberger say that it closely resembles P. cauliorchis;
this is fully borne out by comparing the two descriptions. There is this
difference, however, viz., that in P. cauliorchis they state that Laurer’s
canal “ opens slightly to right or left of the median line, 60 to 3204 cephalad
of the excretory pore,’ whereas in P. crassum Laurer’s canal is described |
as opening ‘ about 0-72 mm. caudad of the excretory pore.’ But there is
some doubt as to the correctness of their observations in P. cauliorchis for
two reasons. First, in their fig. 62, which is a side view of P. cauliorcis,
they show Laurer’s canal running parallel with the excretory canal, and
although the termination of Laurer’s canal is not actually shown, it looks
as if it would open about the same level as the excretory pore. In the
second place, they have made a subgenus Cauliorchis of which P. cauliorchis
is the type, and in their definition of this Stiles and Goldberger’state that
Laurer’s canal is ‘ caudad to caudo-laterad of the excretory pore, the two
pores may be close together . . ... Another point is that in P. crassum the
testicular fields are said to overlap and in P. cauliorchis they are said to be
separate, in the latter case being similar to their definition of P. calicophorum.
The writer has already shown, and is confirmed by Fischoeder, that in
P. explanatum the testes are separate in young worms, and that in older
ones the fields overlap, and although age is the most important factor in
influencing this difference, the writer has found young worms of the same

143

type with the testes overlapping. It is accordingly concluded that there
is no difference between P. crassum and P. cauliorchis.

Paramphistomum fraternum, Stiles and Goldberger, 1gto.

Railliet, Henry and Bauche (1914a) state that P. fraternumis a synonym
of P. explanatum, and they advance such good reasons for their opinion
that it is proposed to accept: their conclusion without further discussion.
In the light of the present investigation, however, there is one point of
interest in the conclusion of Railliet, Henry and Bauche, viz.: Stiles and
Goldberger give as one of the distinguishing characters of P. fraternum that
the genital pore is opposite the anterior sucker in this species, whereas it is
posterior to the gut fork in P. explanatum. From this it may be inferred
that the above observers do not consider the position of the genital pore
of any specific value. Moreover, Fischoeder also seems to be of the same
opinion, as Railliet, Henry and Bauche state that they obtained his
opinion on their material and that he agreed with them as to its identity.

Paramphistomum siamense, Stiles and Goldberger, 19gto.

This species is differentiated from P. cerv1 and P. fraternum by Stiles and
Goldberger and is therefore considered new by them. In their diagnosis
from P. cervi they use the difference in size of the posterior sucker, the
position of the genital pore, and the shape of the worm, but they omit the
one real distinguishing character, viz., the different relations of the testes
to one another in the two species. P. fraternum is distinguished from
P. siamense because the worms are said to be of different shape; the
‘posterior sucker of P. fvaternwm is relatively smaller than it is in
P. siamense ; and the genital pore is proportionately nearer to the anterior
end in P. fraternum. The question of small differences in shape may be
disposed of by reference to Plate VI, fig. A, which is a photograph of
several specimens of P. explanatum, all of which were taken from one
bottle. With regard to the position of the genital pore, this is given as
midway between that described for P. explanatum and P. fraternum, so the
opinion of Railliet, Henry and Bauche (1914a), which has been quoted
under the discussion of the latter species, is of equal value in this case.
The relative difference in size of the posterior sucker in the two species
requires more careful consideration.

144

In the case of P. fraternum Stiles and Goldberger say that only two
specimens ‘in poor condition ’ were available. The length of one of these
specimens is given as 9°75 mm. when in alcohol, and the same worm
measured 8-94 mm. after imbedding and sectioning. The diameter of the
posterior sucker of this worm is given as 3-75 mm. in the antero-posterior
and 3-25 mm. in the transverse direction. In the case of P. stamense the
length of two specimens measured in glycerine alcohol is given as about
6 mm. and g mm. respectively. The posterior sucker was measured in
three “ press preparations, and the diameters of the three are given as
3-5 mm., 4 mm. and 5 mm. respectively. It is not quite clear what is
meant by press preparations, but it is assumed that it means that the
worms were subject to pressure before being measured. The writer has
found that pressure between slides is of great advantage in obtaining a
clear view of the internal organs in carbolic cleared specimens, but it was
found that measurements of worms while under pressure in this way
were quite unreliable. This was especially the case with regard to the
posterior sucker ; because, the worms being much thicker at the posterior
end than at the anterior end, when pressure was applied a considerable
flattening and consequent distortion was caused to the hinder end before
the anterior end was subject to any pressure. It is therefore clear that the
posterior portions of the worms were always relatively more distorted than
the anterior portions, and hence the structures in the hinder part were |
relatively greatly increased. It seems that this is what has occurred in
Stiles and Goldberger’s case, and that owing to their failure to recognise
this fact they have been led to ascribe specific value to what is in reality
an artificial condition. Indeed, from the writer’s experience, it is surprising
that the difference in size of the posterior sucker in P. fraternum and
P. stamense was not greater than they actually found it. It is therefore ©
considered that the small difference between the relative size of the posterior
suckers in these two worms should not be regarded as of any diagnostic
value, and in consequence the two worms are probably identical with one
another and with P. explanatum.

As a result of the above investigation it is concluded that P. calico-
phorum, P. crassum, P. cauliorchis, P. fraternum and P. siamense are all
synonyms of P. explanatum.

In all the species of the genus Paramphistomum that have been dealt
with up to the present the excretory pore is anterior to the opening of

145

Laurer’s canal. Those species in which Laurer’s canal opens anterior to
the excretory pore will now be considered. As the writer has not had an
opportunity of examining the following species, the description is in each
case summarised from the original.*

Paramphistomum orthocoelium, Fischoeder, rgor.

SYNONYMY :—

Paramphistomum dicranocoelium, Fischoeder, 1901.
Paramphistomum streptocoelium, Fischoeder, 1901.
Paramphistomum scoliocoelium, Fischoeder, 1904.
Paramphistomum parvipapillatum, Stiles and Goldberger, 1910.
Parampbhistomum shipleyt, Stiles and Goldberger, 1910.

First found in the stomach of a Bos kerabau that died in Berlin.

The following notes were made from Fischoeder (1903).

Cuticular papillae are present on the anterior end of the worm. The
oesophagus is twice as long as the anterior sucker, and is surrounded by
especially large and numerous cells. The muscle walls of the oesophagus
are not thicker than in most other species of the genus. The gut caeca are
not wavy, they are almost straight ; they lie about midway between the
dorsal and ventral surfaces on each side of the worm, and they end 0-5 mm.
to I mm. in front of the posterior sucker; the diameter of the caeca
is I mm. to 1:2 mm. dorso-ventrally and 5004 to 600 transversely.
With the exception of the vitellaria, the genital organs exhibit no special
characters. The genital papilla is well developed and almost fills the
atrium, being in most cases slightly protruded. The testes lie one behind
the other in the posterior half of the worm ; their size and the distinctness
of their lobulation is variable. The vitellaria, however, consist of round to
oval groups of follicles nearly uniform in size and about 3004 in diameter.
As a rule, they are in a single row along the ventral border of the caeca on
each side except near the posterior end, where they are grouped to the
number of from four to six follicle groups which lie close together. The
eggs are 105m to 115m in length by 60m to 65. in breadth.

* Since completion of this paper the writer has been able to examine a collection of a few
specimens of P. orthocoelium from the stomach of a sheep at Hong Kong. Although it is necessary
to section the worm to see the course of Laurer’s canal, simple clearing in carbolic acid is sufficient
to establish the identity of this species. It is easily distinguished from P. cervi by the vitellaria which
in this species are arranged in very numerous comparatively small groups of follicles, never more
than about 300 in diameter, and showing a marked tendency to encroach on the dorsal and ventral
surfaces of the worm, whereas in P. orthocoelium the number of follicle groups is much less, they are
considerably larger, being up to 7ooy’ in diameter, and are limited almost exclusively to the lateral
fields external to the gut caeca. :

146
Paramphistomum dicranocoelium, Fischoeder, 190t.

The following notes are taken from Fischoeder (1903).

Cuticular papillae were not seen on the anterior end. The worm is
very near P. orthocoelium. The oesophagus is shorter than in P. ortho-
coelium, its extreme length being one and a half times as long as the
anterior sucker, and instead of becoming thicker only quite close to the gut
fork the muscle wall commences to thicken about its middle, and from
this point it gradually increases from 204 to 60-75 at the posterior end.
The gut caeca are straight, but differ from P. orthocoeliwm in being nearer
to the dorsal than to the ventral surface, and they are only 250m to 3504
in diameter; they end about I mm. in front of the posterior sucker.
The genital papilla is well developed, but is strongly retracted as a rule.
The vesicula seminalis is larger and lies farther towards the dorsal surface,
but on the other hand the pars musculosa is not so long as it is in
P. orthocoelium, and the pars prostatica is somewhat shorter also. The
ductus hermaphroditicus may be a fairly broad pear-shaped cavity, or,
when the papilla is strongly retracted, the ductus hermaphroditicus may be
merged in the genital atrium. The vitelline glands are arranged in similar
groups to P. orthocoelium, but they lie as a rule in two rows and show no
special grouping behind the ends of the caeca. The anterior ends of the
vitellaria may be at different levels. The other genital organs are
approximately the same as in P. orthocoelium. The eggs measure 1454
to 150m in length and 75 to 80u in breadth.

Paramphistomum streptocoelium, Fischoeder, 1901.

The following notes were taken from Fischoeder (1903).

Cuticular papillae are present on the anterior end as in the case of
P. orthocoelium. The oesophagus is only about as long as the anterior
sucker ; the gut caeca are strongly convoluted and end opposite the base
of the sucker. The genital atrium is distinguished by the presence of a
ring-like prominence on its inner wall which divides the atrium into
a small ventral and a large dorsal chamber. The testes, as well as varying
in proportion to the size of the worm, may be of different sizes in the
same worm and apparent partial atrophy of these organs has been noted in
gravid worms. The position of the testes is the same as in P. dicranocoelium.
The female genitalia only show slight differences from the previous species.

147

The vitellaria are confined to the sides of the worm and stretch from the
gut fork in front to the posterior sucker behind as in P. dicranocoelium.
The follicles are in groups similar to the previous species (300 to 700” in
diameter), but they are present in larger numbers and are therefore
smaller in size than in the two previously named species. The anterior
limits of the vitellaria may be at different levels on the two sides as in
P. dicranocoelium. The eggs measure 105 to 115 in length by 604 to
65 in breadth.

Paramphistomum scoliocoelium, Fischoeder, 1904.

The following notes are taken from Fischoeder (1904).

The musculature of the oesophagus is similar to that of P. dicranocoelium.
The gut caeca are less wavy than in P. streptocoelium, and they are nearer
to the dorsal than the ventral surface as in P. dicranocoelium. The genital
pore is, as a rule, widely open. The vitellaria are similar to the other
three species ; they are composed of coarse follicles which lie at the sides
of and external to the gut fork, from about the level of the gut fork in front
opposite to the base of the posterior sucker behind. The eggs are 1354 to
145 in length by 65 to 75m in breadth.

Paramphistomum parvipapillatum, Stiles and Goldberger, 1910.

The following notes are taken from Stiles and Goldberger (1910).

The cavity of the anterior sucker is furnished with moderate sized
papillae. The oesophagus is estimated to be not shorter than the anterior
sucker and it exhibits a thickening of its muscle wall in its posterior half.
The gut caeca are slightly wavy and end opposite the middle of the
posterior sucker. The genital papilla is embraced by a ring-like collar
beset with fine papillae. External to this collar is another ring marked off
from it by a groove, and this groove may not be present in cases with the
genital papilla protruded. The vitellaria consist of numerous follicles
lying at the sides of, above, and below the gut caeca ; the gland groups on
the left-hand side extend slightly further inwards than those on the
right hand. The vitellaria extend from the hinder end of the anterior
sucker in front to opposite about the middle of the posterior sucker behind.
The eggs measure 135 in length by 674 in diameter.

148

Paramphistomum shipleyi, Stiles and Goldberger, 1910.

The following notes are from Stiles and Goldberger (1910).

The entrance to the anterior sucker and the sucker itself are lined with
small papillae. The muscle wall of the oesophagus begins to grow thicker
from about the middle of its length towards the posterior end. The caeca
are wavy and end about the level of the anterior border of the cavity of the
posterior sucker. The diameter of the gut varies at different points
along its course, there being marked dilatations followed by constrictions.
The testes are one behind the other, but overlap slightly. The ductus
ejaculatorius opens close to, but separate from and just above the opening
of the uterus. The space into which this ductus ejaculatorius opens is
narrow and slit-like.

‘ From this space a short duct passes ventrad and may be regarded as piercing the
axial region of a mushroom-like structure (figs. 123-126) to open into another slit-like
atrium somewhat larger, however, than the one into which the male and female
ducts open. A duct about 30m in diameter leads from this atrium and apparently

pierces a stout conical papilla, which may be regarded as the genital papilla, to
open into a small genital atrium which connects with the exterior by the genital pore.’

The eggs are about 135 in length by 71 in breadth.

In conducting a critical examination into the specific value of the
above worms, it is in the first place obvious that the points used for
differentiation are all similar to those used in the group of worms that have ~
been considered under P. cervi. But in the present instance the differences
are even smaller than in the former case. For instance, differences in
length of the oesophagus greater than in these six species have been noted
in all species examined. This is only to be expected when it is realised
that the oesophagus is well supplied with muscle, contraction or relaxation
of which can easily account for the differences observed. The amount of
increase of thickness towards the posterior end of this organ has also been
shown to vary considerably in the same species. The point of termination
of the gut caeca only varies very slightly in the above six species ; this
character has been found to have no diagnostic value in other cases, and that
in the present instance it is equally valueless is appreciated, when it is
noticed that in P. scoltocoelium, Fischoeder shows the caeca ending in
front of the posterior sucker in fig. 7, and behind the anterior border of the
sucker in fig. 8, although in the text he says they end in front of the
posterior sucker. It has also been found that slight differences in the

149

amount of convolution of the caeca, as well as in their diameter, are of no
value. A moment’s reflection will explain the reason for this. The caeca
are hollow structures furnished with muscular walls and it will thus be
obvious that the amount of convolution may vary considerably according
to the degree of contraction of the muscular wall of the gut and of the
whole worm. The value of slight differences in the distribution of the
vitellaria has also been shown of no use for specific diagnosis in all other
species, and that the same probably applies in the present series of worms
is borne out by comparison of the statements in regard to these glands in
several of the worms. For example, Fischoeder states that as a rule the
gland groups are in a single row in P. orthocoelium, and in P. dicranocoelium
they are as a rule arranged in a double row; but as he does not state
what the exceptions to these rules are, all that one can infer is that the
vitellaria are subject to some variation in both these species and cannot,
therefore, be accorded any specific value. The lability to variation of the
vitellaria is also borne out by the following contradictory statements by
the same author: in P. orthocoelium he says the vitelline follicles are in
groups, each of which is about 300 in diameter ; in P. dicranocoelium they
are similar to the above ; and in P. streptocoelium they are in groups similar
to the above, but having a diameter of from 300u to 700; whilst in the
next sentence he states they are present in P. streptocoelium in far greater
numbers than in the two previous species and are in consequence of
smaller size. With regard to the genital apparatus, there is no evidence of
any allowance having been made for variations due to protrusion or retrac-
tion of.the genital papilla at the time of fixation. for example, the genital
papilla of P. orthocoelium is described as being well developed and in most
cases filling the atrium ; in P. dicranocoelium the papilla is said to be well
developed and in most cases strongly retracted; in P. streptocoelium
a ring-like prominence divides the atrium into two chambers; in
P. scoliocoelium the genital pore is stated to be, as a rule, widely open ;
in P. parvipapillatum, the papilla is described as being embraced by
a collar-like ring beset with minute papillae ; and in P. shipley: from the
single sectioned example that they studied Stiles and Goldberger describe
a complicated arrangement of ducts and atria which, in the writer's opinion,
is only a description of a worm with the papilla in strong retraction such as
can be found in any species, if enough specimens are examined. All the
various descriptions of genital papilla and atrium described above have
been seen by the writer in all the species in which he has had sufficient

150

material to examine a long series of mature worms. It is therefore
considered that in the present case they cannot be regarded as specific
differences. The only other point relates to the small papillae described
as present in some species of this group of worms and absent in others.
In all other species they have been found to vary to a great extent, and
accordingly it is probable that in the present case they vary in the same
way, and are in consequence of no diagnostic value.

The evidence obtained from the examination of closely allied species,
indicates that the points on which the differentiation of the above six species
is based, are of no specific value. It is therefore considered that
P. dicranocoelium, P. streptocoelium, P. scoliocoelium, P. parvipapillatum
and P. shifleyi are synonyms of P. orthocoeliwm.

Paramphistomum buxifrons, Leiper, 1910.

Host :—Hippopotamus sp. Location :—Stomach. Locality :—Uganda.

According to Leiper, P. buxifrons can readily be distinguished by its
leaf-like shape which resembles the leaf of a box tree. A large example
may measure 5 mm. in length by 3 mm. in breadth and be only 0-4 mm. in
thickness at the middle of the body. The testes also are a distinguishing
character, as they are not lobed and they are placed diagonally in the
posterior part of the worm. The only other species with testes not lobed
are P. liorchis and P. wagandi and in these cases the testes are one behind
the other, but in P. buxifrons, Laurer’s canal opens in front of the excretory
pore, whilst in P. liorvchis Laurer’s canal is behind the excretory pore.

Paramphistomum wagandi, Leiper, 1910.

Host :—Hippopotamus sp. Location :—Stomach. Locality :—Uganda.

The testes are not lobed and are placed one behind the other, and the
worm is therefore very similar to P. liorchis; the two species are
distinguished by the position of the opening of Laurer’s canal, which is in
front of the excretory pore in P. wagandi, whereas it is behind the excretory
pore in P. liorchts.

In his descriptions of these species, Leiper lays down by definite
measurement the exact size and position of the various organs, the appear-
ance of the genital atrium, the shape of the excretory bladder, etc. In most
cases these particulars appear to have been obtained from a single sectioned

151

specimen. This fact is considered unfortunate, because slight variations
from the data given by Leiper are almost certain to be found when these

worms are more fully known, and will in all probability lead to a multiplica-
tion of species on the ground of slight variations, in the same way as seems
to have occurred in most of the other species.

Genus Colylophoron, Stiles and Goldberger, rgrto.

Definition.—Paramphistominae: with a genital sucker distinctly
marked off from the subcuticular muscle layer.

Type species : Cotylophoron cotylophorum (Fischoeder, 1gor), Stiles and
Goldberger, 1910. |

Key To SPECIES

Laurer’s canal opens posterior to excretory pore ... ae ... C. cotylophorum
Laurer’s canal opens anterior to excretory pore... oe Je C. minutum

Cotylophoron cotylophorum (Fischoeder, 1901), Stiles and Goldberger, IgIO.
SYNONYMY :—

Paramphistomum cotylophorum, Fischoeder, 1901.
Cotylophoron indicum, Stiles and Goldberger, 1910.

First found in the stomach and intestines of Bos sp. in German East
Africa.

The material available for study in the present case consisted of the
following collections :— |

1. ‘Ten bottles from the stomach of bullocks killed at Sierra Leone, West
Africa.
2. Four bottles from the stomach of buffaloes (Rubales sp.) in the Upper
Shire River, Nyasaland.
3. Four bottles fromthe: stomathtofitixe inswala (Aepyceros melampus) shot
in the Upper Shire River District, Nyasaland.
4. One bottle from the stomach of a Pagan dwarf bull from Ilorin, Northern
Nigeria.
One bottle from the stomach of a waterbuck (Cobus sp.) from Zeref,
Khartoum.
One bottle from the stomach of a hartebeest (Bubalts sp.) from Nyasaland.
One bottle from the stomach of an antelope sp. (?) from Rhodesia.
One bottle from the stomach of an antelope sp. (?) from Nyasaland.

oo OMe

In some of these bottles were many hundreds of specimens, so ample
material was available for examination.

The species C. indicum was made by Stiles and Goldberger from six
specimens. They state that no eggs were observed, so it is probable that
their material was immature.

152

The differences between C. cotylophorum and C. indicum are summarised
by Stiles and Goldberger as follows :—

‘ Cotylophoron indicum comes close to C. cotylophorum from which it differs
chiefly in the structure of the oesophagus, which is provided with a bulbous thickening
in the latter species, but is without it in the former. The two differ also in the
details of structure of the copulatory apparatus and in the position of the genital
pore. In C. indicum the genital sucker is less sharply delimited, projects less, has
a much smaller genital atrium, and the genital pore is decidedly postbifurcal ; on
the other hand, in C. cotylophorum the genital sucker is sharply marked, with rim
prominently bulging the venter, with a relatively roomy genital atrium and with
the genital pore in the bifurcal zone.’

It should be noted that Fischoeder (1903) in his description of
P. cotylophorum says the uterus is strongly convoluted and it is full of eggs,
showing that his specimens were mature.

The following points have been worked out from the examination of
many specimens of C. cotylophorum, either cut and mounted in serial
sections, or examined whole in carbolic acid.

Oesophagus. In their description of the species C. indicum, Stiles and
Goldberger state that the walls of the oesophagus are thick, but give no
other important details of its characters. In the present instance, the
thickness of the muscle walls of the oesophagus, as well as its length and
direction, were found to be very variable. Although it was much more
distinct in some cases than in others, there was always a gradual increase
in thickness of the muscle wall of the oesophagus from the anterior end
towards the posterior end, in exactly the same way as described in P. cervt.
Fig. 7 represents camera lucida drawings of eleven specimens of
C. cotylophorum cut in sagittal section. It will be noted that in figs. K and A,
the oesophagus is approximately the same as in fig. 45 by Stiles and
Goldberger, which is a drawing of C. indicum. Now, taking the remaining
drawings in fig. 7 in the following order D, B, E, C, L, F, G, H, it will be
observed that as the oesophagus gradually increases in length, its posterior
extremity becomes more and more bulbous. Drawings C, L, G, and F are
very similar to Fischoeder’s fig. 38, which is a drawing of C. cotylophorum
cut in the sagittal plane. In addition to the above characters it was
noted that the worms from which drawings J and K were made contained
no eggs, whilst all the others had eggs in the uterus, those with the longest
oesophagus having the most eggs. It seems probable, therefore, that the
differences in the oesophagus in C. indicum and C. cotylophorum are
really due to differences of age. This is all the more likely when it is
remembered that Stiles and Goldberger’s material did not contain eggs,

153

and that Fischoeder’s did. Two other points are also well illustrated in the
above series of drawings, viz., that the course and length of the oesophagus
are subject to considerable variation, and that the position of the genital
pore varies in relation to the gut fork to such an extent that neither of
these points is reliable for distinguishing between C. indicum and
C. cotylophorum.

Fic. 7. Cotylopboron cotylophorum. Sagittal section through the anterior ends of eleven
specimens to show oesophagus and genital sucker. a.s.—anterior sucker; g.p.—genital pore ;
g-5.—genital sucker ; 1.—intestine ; 0.—oesophagus. 12.

Genital apparatus. The differences in size, and degree of extrusion or
retraction of the genital sucker and papilla, with consequent changes in
the genital atrium, were almost as numerous as the specimens examined.
This is well illustrated in figs. 8 and 9, which show such great differences in
appearance that at first sight it might be considered that different species
are being dealt with, but it should be borne in mind that the above examples
have been chosen for this very reason, and that many specimens have been
examined showing all possible intermediate stages, thus rendering it
practically certain that it is only a question of individual variation.
Fig. 8, Ar and A2, are two sections of the same worm, and they have been
included because Fischoeder (1903) states that the male and female

154

openings have always been noted by him to be separate in C. cotylophorum
as shown in fig. 8, A2, which closely corresponds to Fischoeder’s fig. 38.
But a later section (fig. 8, Ar) showed the male and female ducts uniting
within the substance of the genital papilla and opening on the surface by

\ ae yy,
AAA \ 1}
: \\ \M 4 5 \

y de.
ih ut.

——
—S ee
—

Fic. 8. Cotylophoron cotylophorum. Sagittal section through genital sucker. 41—Section
showing tip of genital papilla with male and female ducts uniting in the usual way. 42—Section
of same specimen with male and female ducts apparently opening separately, because tip of genital
papilla is not seen. B—Section of another specimen with genital papilla lying in a wide atrium.
d.e.—ductus ejaculatorius ; d.b:—ductus hermaphroditicus ; g.a.—genital atrium; g.p.—genital ~
pore; g.pp.—genital papilla; g.s.—genital sucker ; ut.—uterus. X 45.

a common duct. It is therefore clear that there is no essential difference
between this worm and other species with regard to the términation of the
male and female ducts. Fig. 8, B, shows how different the genital pore may
appear if it is relaxed with a large genital atrium and a patulous genital

pore, but in sections not passing through the genital atrium the genital

155
sucker was seen to be just as thick as in other cases. In fig. g two very
different appearances of the genital apparatus are shown. Fig. 9, A, shows
the whole genital sucker protruded beyond the surface of the worm, while
in 9, B, it is deeply retracted within the body. It will be noted that in
fig. 9, A, the subcuticular muscle extends past the base of the genital sucker
in an apparently unbroken column of fibres, and that even opposite its base
the parenchyma is not sharply marked off from the sucker, whereas in
fig. 9, B, with the sucker deeply retracted, this separation is quite distinct.
The conclusions drawn from these two figures are that the demarcation of

Fic. 9. Cotylophoron cotylopborum. Sagittal section of genital sucker of two specimens.
A—Genital sucker fully extruded. x 40. B—Genital sucker fully retracted x 60. cu.—cuticle ;
d.b.—ductus hermaphroditicus; g.a.—genital atrium; g.p.—genital pore; g.pp.—genital
papilla; g.s.—genital sucker; p.—parenchyma; p.m.—pars musculosa; /.p.—pars prostatica ;
s.m.—sphincter muscle; w#f.—uterus.

the sucker from the parenchyma in this genus is more apparent than real,
and that it is probably brought about by the subcuticular muscle and the
contiguous parenchyma being stretched around the sucker when it is in the
retracted condition. Comparison of the shape and size of the atrium in the
drawings will make it sufficiently clear without further comment that
minute descriptions of the shape and number of chambers in this atrium
cannot be regarded as of any value for specific diagnosis. The conclusion
arrived at from consideration of the above facts is that Stiles and
Goldberger, in describing C. indicum from six specimens with no eggs in the
uterus, were in all probability dealing with immature specimens of
C. cotylophorum.

In addition to the above points, which were worked out in sectioned

156

specimens, evidence of variation in other characters was obtained by the
examination of eighty specimens cleared in carbolic acid and examined
whole.

Gut caeca. The amount of convolution of the caeca varies considerably,
as does their point of termination. The most usual position for the caeca
to end in this species is about opposite the middle of:the posterior sucker,
but in some cases they both ended well in front of the anterior border of
the posterior sucker, and in others they nearly reached the posterior border
of the posterior sucker. Between these two extremes all intermediate
stages were found, and in a few instances the caeca on the two sides of
the same worm ended at different levels. As a rule, the final turn of the
caeca was directed dorsally, but this was by no means invariable, as is
clearly shown in fig. Io.

Dorsal.

oO \ wy
ant. (NO) 5 irs | Re Wy Post.

[Ww BS lee >W]

Fic. 10. Cotylophoron cotylopborum. Drawings showing various forms of termination of
gut caeca. ‘These figures are not to scale, and are drawn from carbolic cleared specimens. Those
in brackets are the two caeca from a single worm.

Vitellaria. The distribution of the vitelline glands and the number of
collections of follicles in each gland showed a remarkable degree of
variation. The glands commenced anteriorly anywhere from about opposite
the middle of the anterior sucker to a point slightly behind the genital pore ;
as a rule, they extended posteriorly to a little behind the termination of the
gut caeca, but in a few instances groups of follicles were seen extending to
the extreme posterior end of the worm and surrounding the opening of the
posterior sucker in the same way as in all the other species examined.
The degree of extension inwards on the dorsal and ventral surfaces varied
in the same way as in P. cervi. The anterior extension of the gland was
often different on the two sides, and in one extreme instance the vitelline
gland on the left side began in front of the genital pore and ended just

behind the corresponding caecum, but on the other side the gland was
composed of a closely packed collection of follicle groups, which commenced
opposite about the middle of the hinder testis and ended opposite the
middle of the posterior sucker, being completely confined to the outer side
of the caecum of that side.

Excretory system. The change in the relations of the excretory pore to
the excretory bladder in worms of different age, which has been referred
to in P. cervt and P. explanatum, has been found to hold good in the case of
C. cotylophorum. ‘This is clearly shown in fig. 11, which represents the

Fic. 11. Cotylophoron cotylophorum. Sagittal sections of three specimens near the mid-line
to show the alteration in course of the excretory canal with increasing age. A—Immature worm.
B—Partly gravid worm. C—More fully gravid worm. ex.b.—excretory bladder;
ex.p.—excretory pore; JL.c.—Laurer’s canal; .s.—posterior sucker; sh.g.—shell gland;
ut.—uterus x 18.

posterior ends of three specimens of C. cotylophorum of different ages.
These figures also emphasise the fact that the shape of the excretory
bladder is very variable.

Leiper (1910) described two worms from the hippopotamus, which he
placed in the genus Pavamphistomum, but both of them have a definitely

158

delimited muscular thickening round the genital pore. Following the
classification of Stiles and Goldberger (1910) these worms should be placed
in the genus Cotylophoron. The following are the principal distinguishing
features of these species according to Leiper’s descriptions.

Cotylophoron minutum (Leiper, IgI0).
Synonymy :—Cotylophoron sellst (Leiper, 1910). |

First found in the stomach of a Hippopotamus sp. in 1 Uganda.

The testes are small and oval in shape and they lie one behind the
other, being separated by a space about equal to the diameter of a single
testis. Although it is not stated in the text, it is assumed from Leiper’s
fig. 32 that the testes are not lobed. Laurer’s canal opens in front of the
excretory pore. This worm differs from C. cotylophorum in that the testes
are not lobed and that Laurer’s canal opens in front of the excretory pore.

Cotylophoron sellst (Leiper, I9ro).

This species is very similar to C. minutum, the only differences being
that it is larger and that its testes are relatively larger and closer together
than in C. minutum. Leiper also describes a large genital atrium, but as he
says that he made his observations from a single sectioned specimen this
cannot be regarded as of specific importance.

The only differences of importance therefore between the two species
C. sellst and C. minutum are those of size, and as quite as big a range of —
variation has been found in several series of C. cotylophorum examined by
the writer, both with regard to the size of the whole worms and of the testes,
it is considered probable that C. minutum and C. sellsi are identical.

Sub-family CLADORCHINAE, Fischoeder, Igor.
Definition.—Paramphistomidae : oral sucker with a pair of diverticula.

Key To GENERA

1. Cirrus pouch absent sv al vate ee ant uN Pseudodiscus
Cirrus pouch present... p48 Hy 4% Y ae ap Sa
2. Testessmooth ... — ag: nes SiS eae Balanorchis
Testes lobed or preieeal ~s th ~ de! a: a
3. ‘Testes lobed ma +7 tg Su see Bes “ibs Pfenderius
‘Testes branched . Sse ar Ae pe an
4. ‘Testes each consist inisou branches ike a cross ait +] ... Chiorchis

Testes consist of many long irregular branches
5. Main portion of uterus posterior to testes, terminal portion runs ventral to
Lee oe Ghee a LTaxorchis
Main portion of uterus ‘archés dorsally « over testes, terminal portion anterior to
testes (as in Paramphistomum) AIMEE Wi she Cladorchis

159
Genus Watsonius, Stiles and Goldberger, rgro.

Careful comparison of the definitions of Watsonius and Pseudodiscus,
as given by Stiles and Goldberger, reveal little difference between the two.

In the definition of Psewdodiscus they state the genital pore is behind the
gut fork; a ductus hermaphroditicus is present; the oral sucker is
‘prominently constricted at equator’; the oesophagus has no muscular
thickening ; and the testes lie side by side near the middle of the worm and
are ‘ cauliflower-like,’ which means lobed with lobular subdivisions.

The same points in Watsonius are described as follows :—The genital
pore is in front of the gut fork ; a ductus hermaphroditicus is apparently
absent ; no mention is made as to whether the oral sucker is constricted
or not; the oesophagus has a distal thickening of the muscle layer ; and
the testes are lobulate lying almost one behind the other. All the other
points cited are the same in the two genera, or else are not of any value.

It is repeatedly shown in the present paper that the position of the
genital pore in front or behind the gut fork is not even of specific value.
As the constriction of the oral sucker is only alluded to in one genus, this
point may be discarded. The differential diagnosis thus rests on the
apparent absence of a ductus hermaphroditicus, the presence of an
oesophageal thickening and the tandem arrangement of the testes in
Watsonius, against the presence of a ductus hermaphroditicus, the absence
of an oesophageal muscular thickening, and the lateral position of the
testes in Pseudodiscus. In the sub-genus, Hawkesius of Pseudodiscus,
however, Stiles and Goldberger state that there is a pronounced thickening
of the posterior part of the oesophagus, and the testes are tandem. In
respect of these two characters, the definition of Pseudodiscus as given by
them cannot be correct. Thus, all the differential points between the
genera Pseudodiscus and Watsonius are eliminated, except the doubtful
absence of a ductus hermaphroditicus in the latter. The fact that Stiles
and Goldberger in erecting the genus Watsonius had only a single specimen
cut in transverse section, which they borrowed from Shipley, makes this
point of very doubtful value, because it is so difficult to make out a ductus
hermaphroditicus in sections of this nature, that it is unsafe to rely on them
without confirmation from examination of sagittal sections. Leiper (1913),
however, in a short record of W. watsoni figures the ductus ejaculatorius and
uterus opening separately on the end of a prominent papilla, there being
a narrow tongue-like process between the two ; thus he apparently confirms

160

Stiles and Goldberger’s statement that a ductus hermaphroditicus is absent.
In the same paper, Leiper gives a drawing of Gastrodisco1des hominis in which
the genital papilla and the two ducts are shown to be practically identical
with his figure of W. watsont ; the writer has examined sagittal sections of
three specimens of G. hominis and found a papilla with a long ductus
hermaphroditicus in the first specimen (fig. 29), a much shorter ductus
hermaphroditicus in the second, and in the third there was no papilla at all
and the male and female ducts opened separately at the bottom of a deep
atrium (fig. 28). It seems probable that the same variation could be found
if a number of W. watsont were examined ; this is all the more likely when
it is remembered that the same range of variations has been found in
every species which the writer has been enabled to examine.

This discussion indicates that there are no clear differences between
the genera Pseudodiscus and Watsonius, and that the latter should be
merged in the former.

It should be noted that Railliet, Henry and Joyeux (1912), in recording
Watsonius watson from Cercopithecus callitrichus, draw attention to the
great similarity between this worm and Hawkesius, the sub-genus of
Pseudodiscus created by Stiles and Goldberger (1910).

Genus Pseudodiscus.*
Synonymy :—Watsonius, Stiles and Goldberger, 1910.
Definition.—-Cladorchinae : without a cirrus pouch, and testes lobed.

Type species Pseudodiscus collinsi (Cobbold, 1875), Sonsino, 1895.

Key To SPECIEs
Testes side by side... on: 2 a ae be oh: P. collinst
Testes tandem Lae ae ae _s ee P. hawkesti and P. watsoni

Pseudodiscus collinst (Cobbold, 1875), Sonsino, 1895.
SyNonyMy :—Pseudodiscus stanleyi (Cobbold, 1875), Sonsino, 1895.

First found in the colon of the horse in India.

The material investigated all came from India, and was as follows :—

* Stiles and Goldberger divide the genus Pseudodiscus into two sub-genera, viz., Pseudodiscus,
and Hazwkesius, but as this serves no useful purpose, it is not used in the present paper.

161

Bottie 1. Four specimens from a pony. ‘These were all about the same size,
viz., 7mm. long and § mm. broad, and none contained eggs.

Botte 2, Three specimens from a pony. ‘I'wo of these were about 7-9 mm.
long by 5:4 mm. broad, and both had eggs in their uteri. The third specimen

was 5:7 mm. long by 4:4 mm. broad, and had no eggs.

Bottte 3. ‘Three specimens from a horse. All three were very small, the
largest being only 2-9 mm. long by 1-3 mm. broad.

Bottte 4. One specimen from a mule. About medium size.

Borrie 5. ‘Three specimens from a horse. They were all small and averaged
4:3 mm. long by 2 mm. broad, and were all obviously immature.

Borie 6, Sixty-four specimens from a horse. The maximum length of any
of these worms was 4°8 mm. and the maximum breadth 2:5 mm., the minimum
length was 3:2 mm. and the minimum breadth 1-3 mm., with all gradations between
these two dimensions. But the longest worm was only 2:1 mm. broad, and the
broadest worm was only 4:7 mm. long, therefore, the proportion of length to breadth
is not absolute. ‘The average of the whole sixty-four specimens was 4:4 mm. long
by 1-9 mm. broad,

Borrte 7. Thirty-five specimens from a horse. ‘These worms varied between
8-7 mm. long by 5-7 mm. broad and 5-4 mm. long by 3:2 mm. broad, and several
of the larger specimens contained eggs.

(Bottles 6 and 7 were kindly lent by Mr. A. W. N. Pillers, |e C. VD),
For convenience, the points in which the two worms P. stanley1 and
P. collinsi difter, according to Stiles and Goldberger (1910), are arranged in

tabular form (Table VII). |
Tasre VII.

2. ‘Testes. Actually and relatively smaller. | Actually and relatively larger.

3- Opening of Laurer’s | A little above the posterior border of | A little behind the anterior border

canal. the acetabulum. | of the acetabulum.

4. Size of acetabulum. Relatively larger. Relatively smaller.

5. Position of ovary About level of upper margin of | Distinctly further forward and
and shell gland acetabulum. nearer testes.

It must be noted, however, that Stiles and Goldberger record that
they had only a few specimens of each of the two worms and, as they state
that no eggs were seen, it is doubtful whether their worms were fully
developed.

162

On looking through the Liverpool material, a doubt was raised in one’s
mind whether the differences described by Stiles and Goldberger were
really of specific value, as it appears from our material that all inter-
mediate stages between the two extremes were to be seen. With a view to
examining this more fully, Table VIII was compiled from data obtained
from Stiles and Goldberger (1910), from measurements of their drawings,
and also from those of nine specimens taken from bottles 1, 2, 3, 6 and 7 of
the Liverpool material, as far as possible representing all sizes of the worms.

Comparison of the dimensions given in the table makes it clear that
Stiles and Goldberger’s differences referring to the relative size of the
oesophageal portion of the sucker, and the relative size of testes and
acetabulum, are merely due to the stage of development of the various
specimens that they examined. In all trematodes the digestive and
fixation organs develop earlier than the sexual glands ; therefore, it is to be
anticipated that the former are relatively larger and the latter relatively
smaller in young worms than in older specimens. The differences of
position of the ovaries and openings of Laurer’s canal are very minute,
and the above authors had so little material available, that to give specific
value to slight variations in positions of these organs, as they have done,
does, not appear to be justified. |

Historical. Cobbold (1875a) examined thirty-three specimens of this
worm from a horse in India, which were sent him by Collins, and he
named them Amphistoma collinsi because they were smaller than a similar.
fluke of the elephant (A. hawkesi) from the same locality. A few days
after publication of this paper he received from Professor Simonds another
bottle of flukes from the horse, collected by Stanley in India, and in a paper
(Cobbold, 1875b) he named them Amphistoma stanleyi, but in doing so,
says ‘ This is apparently nothing more than a large variety of the above ? ’
He made no detailed study of the worms in either case, and no adequate
description of the internal anatomy was published until Stiles and
Goldberger (1910) examined some of Cobbold’s original material. The
number of individuals they had for study was not large, therefore they were
led to making specific characters of differences which are really due to
differences in age of the specimens they examined.

Pseudodiscus stanleyi (Cobb., 1875) is therefore only an immature form
of Pseudodiscus collinsi (Cobb., 1875) and must be regarded as synonymous
with the latter.

163

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164
Pseudodiscus hawkesi1 (Cobbold, 1875), Sonsino, 1895.

In a list of Trematodes from the Asiatic elephant, Railliet, Henry and
Bauche (1914b). give the two following species of Amphistomes :—

(a) Pseudodiscus hawkesi (Cobbold, 1875).

SYNONYMY :—
Amphistoma hawkesi1, Cobbold, 1875, non Piana and Stazzi, 1900.

Pseudodiscus hawkest, Sonsino, 1895.
(b) Watsonius ornatus (Cobbold, 1882).

SYNONYMY :—
Amphistoma ornatum, Cobbold, 1882...
Pseudodiscus ornatus, Sonsino, 1895.
Amphistoma hawkest, Piana and Stazzi, 1900.
Hawkesius hawkest, Stiles and Goldberger, 1910.
Watsonius ornatus, Railliet and Henry, 1912.

Stiles and Goldberger (1910) state that their species Pseudodiscus
(Hawkeswus) hawkesii is a synonym of Amphistoma hawkesit, Cobbold, 1875.
It therefore seems clear that Pseudodiscus hawkest (Cobbold, 1875), and
Watsonius ornatus (Cobbold, 1882) are the same. As Cobbold’s original
spelling was hawkesw and not hawkest the correct name of the worm is
Pseudodiscus hawkesu (Cobbold, 1875). This worm is easily distinguished
from P. collinst, because the testes in P. hawkesit are tandem and in

P. collinsi they are placed side by side.

Pseudodiscus watsont (Conyngham, 1904).

SYNONYMY :—

Amphistoma watsont, Conyngham, 1904.

Cladorchis watsoni, Shipley, 1905.

Gastrodiscus watsoni, Verdun, 1907.

Paramphistomum watsoni, Manson, 1908.

Watsonius watsoni, Stiles and Goldberger, 1910.

Watsonius macaci, Kobayashi, 1915.

This species has been found on two occasions, once in man by Watson
in 1904, and once in a monkey, Cercopithecus callitrichus, by Joyeux

aL OL 2s

165

Comparison of Stiles and Goldberger’s description of P. hawkesii and
P. watsoni show that the two worms are practically identical. The size is
somewhat different, however, P. hawkesii being 3-5 to 5 mm. in length
by 2 mm. to 3 mm. in breadth, and P. watsoni 8 mm. to ro mm. in length by
4mm, to 5 mm. in breadth ; but in P. hawkesii no eggs were seen, whereas
in the single specimen of P. watsoni which these observers examined eggs
were present. The difference in size is therefore probably due to difference
in age. The writer is of the opinion that the two species are identical, but in
view of the facts that he has not been enabled to examine either, and that
one worm comes from man and the other from the elephant, he does not
feel justified in merging them.
3 Kobayashi (1915) recorded Watsonius macaci from Macacus cynomolgus
and the same author (1920) gives a description of the worm, adding a foot-
note to the effect that it is probably identical with the fluke identified as
W. watsoni by Railliet, Henry and Joyeux (1912). If this is correct,
W. macaci is a synonym of P. watsont. ;

Genus Balanorchis, Fischoeder, Igor.

Definition.—Cladorchinae: testes not lobed or branched ; cirrus sac
present and protrusible ; genital sucker present.

Type species Balanorchis anastrophus, Fischoeder, 19ot.

Host :—Cervidae sp. Location :—First stomach. Locality :—Brazil.

Only one species described.

Genus Pfendertus, Stiles and Goldberger, Igto.

Definition.—Cladorchinae: testes lobed ; cirrus sac present ; genital
sucker absent. |
Type species Pfenderius papillatus (Cobbold, 1882), Stiles and
Goldberger, 1910.
_ Host :—Elephas indicus. Location :—Colon. Locality :—India.
- Only one species described.

Genus Chiorchis, Fischoeder, 1901t.

_ Definition. —Cladorchinae ; with testes each consisting of four branches
arranged like a cross; genital sucker absent and cirrus pouch present ;
vitellaria in two narrow rows along outer side of caeca, each follicle group

166

has a duct joining direct with the main longitudinal duct ; uterus as in

Paramphistomum.

Type species Chiorchis fabaceus (Diesing, 1838), Fischoeder, rgot.
Host :—Marine mammals. Location :—small and large intestine.

Locality :—?

Only one species described.

Taxorchis* (Fischoeder, Igo1), gen. nov.

Definition.—Cladorchinae ;_ testes branched; cirrus pouch present ;

genital sucker present ; main portion of uterus posterior to testes, terminal

portion runs anteriorly ventral to testes. t

Type species Taxorchis schistocotyle, Fischoeder, Igot.
Host :—Dictocotyle torquatus. Location :—Caecum. Locality :—Brazil.
Only one species described.

Genus Cladorchis,*t Fischoeder, IQOI.

Definition.—Cladorchinae: with branched “testes, genital sucker

usually present ; cirrus sac present ; uterus asin the genus Paramphistomum.

Type species Cladorchis pyriformis (Diesing, 1838), Fischoeder, 1gor.

Key To SPECIFs

Testes in tandem

Testes side by side... Ly di ny zt «5! ie oe oo

Posterior sucker on ventral surface at
junction of posterior and middle thirds,
small and contains no papillae ... C. pyriformis (Diesing, 1838),
Fischoeder, 1901.
Host :—-Lapirus americanus,
S. America

* Fischoeder classified the species T. schistocotyle under the genus Cladorchis, sub-genus Taxorchis.

But the course of the uterus is so different from any other member of the genus that it is considered
advisable to place this species in a separate genus.

t The uterus arises from the shell gland just in front of the posterior sucker; from this point

it runs forward near the dorsal surface to the posterior border of the testes, it then turns posteriorly
and runs diagonally towards the ventral surface to just in front of the posterior sucker ; after reaching
this point it again turns forward and runs close to the ventral surface to the genital pore.

t Fischoeder divided this genus into three sub-genera, viz., Cladorchis, Stichorchis, and Taxorcbis.

The last has already been dealt with, having been raised to generic rank. Cladorchis and Stichorchis
do not present striking anatomical differences, so they will be considered saad the sub -generic
titles being dropped. O

167

Posterior sucker occupies hinder half of
ventral surface ; large (2 mm.-4 mm.)
and beset with papillae with special
structure a i en ... C. asper, Fischoeder, got.
Host :—Tapirus americanus,
S. America
2. Pharyngeal pouches projecting from
sucker ; genital sucker distinct ... (. giganteus (Diesing, 1838),
Fischoeder, tgot.
Host :—Dzicotyles spp.
Pharyngeal pouches so small that they do
not show on external wall of sucker ;
genital sucker not distinct... ... CC. subtriquetrus (Rudolphi, 1814),
Fischoeder, 1go1.
Host :—-Castor fiber, Bos taurus.

Sub-family STEPHANOPHARYNGINAE, Stiles and Goldberger, 1910.

Definition.—Paramphistomidae : with a single oral diverticulum.
Only one genus.

Genus Stephanopharynx, Fischoeder, Igor.

Definition.—That of the sub-family.
Type species Stephanopharynx compactus, Fischoeder, Igor.
Only one species recorded.

Stephanopharynx compactus, Fischoeder, Igor.

Fischoeder found the worm in Bos sp.

The material available to the writer consisted of the following
collections :—

1. About 850 specimens.
2. Over one hundred specimens.
3. Ten specimens.

These collections were found in the stomachs of three waterbuck
(Cobus sp.) at Ngoa, N.E. Rhodesia.

Fischoeder’s material consisted of two collections composed of a single
specimen in one case and two specimens in the other; they were found
in company with many other Amphistomata.

In addition to the single large oral diverticulum, this worm is charac-
terised by the anterior end being hemispherical instead of the usual conical
type, and its maximum transverse diameter is about its middle instead of

168

towards the posterior end which is the ordinary condition in Paramphis-.
tomidae. The posterior sucker looks ventrally on account of the strong

downward curve which the hinder half of the body exhibits, and it is said

to have a surprisingly sharp border around its opening. The testes lie one

behind the other, the anterior one being about the middle of the body,

with the hinder one between it and the posterior sucker. Laurer’s canal

opens in front of the excretory pore in the mid-line of the dorsal surface,

and the genital pore is surrounded by a thickening of the subcuticular

muscle not sharply marked off from the parenchyma. The size of these

worms is given as 4:8 to 5 mm. in length, 3 mm. in breadth, and 2-5 mm.

in thickness. Other anatomical details are of the usual type. The writer

found in his collection No. I eight worms, and in his collection No. 2 one

worm which agreed with Fischoeder’s description in all particulars, except

that they were about 7mm. inlength. The remaining numerous specimens,
however, at first sight appeared to be very different, as they were sharply
pointed at the anterior end and the posterior sucker looked directly
backwards, in many cases being widely opened and occupying the whole
hinder end of the worm. (Plate VII, fig. B). These worms were of all sizes
from tiny specimens about 2 mm. in length up to worms 5-5 mm. in length.

Several of these worms were sectioned and it was then found that they
agreed with Fischoeder’s species, except that the posterior sucker was
relatively larger, and the hinder of the two testes was much nearer the

ventral surface than the anterior one (fig. 12). At first it was thought that

these points indicated a new species, but it was then noted that none of the

worms were gravid, and prolonged search of the ample material failed to

reveal any specimen containing eggs. On this account it is considered that

they are immature specimens of S. compactus, which fact explains the

relatively large size of the posterior sucker. The apparently different

position of the testes in young worms is not without parallel, for Fischoeder

says that in immature P. calicophorum, the anterior testis lies more dorsal

than the hinder one, a character which the writer has found disappears

when maturity is reached. A further fact in support of the view that the

present worms are only immature S. compactus is that this species has only

been recorded on five occasions (twice by Fischoeder and three times by the

writer) and on two of these it has been found in company with worms of

the above closely allied type. ae

ps. ™ "Cem

Fic. 12. Stephanopharynx compactus. Sagittal section near the mid-line. .a.s.—anterior
sucker; | d.e.—ductus ejaculatorius; ex.b.—excretory bladder; ex.p.—excretory pore ;
g.p.—genital pore ; i.—intestine; Z.c.—Laurer’s canal; 0.—oesophagus; o0.d. i
ov.— ovary ; p.m.—pars musculosa ; p.s.—posterior sucker 3 sh.g.—shell gland ;_ t1.—dorsal testis ;
‘2.—ventral testis; wt.—uterus ;. v.g.—-vitelline gland; v.s.—vesicula seminalis. x 14.

Family GASTROTHYLACIDAE, Stiles and Goldberger, rgto.
Definition —Amphistomata : with a ventral pouch.

Kry To GENERA

1. Uterus crosses from one side of body to the other near the middle of the

worm ... se x Gastrothylax
Uterus lies in centre of body for its whole length tr ap cnar 2

2. ‘Testes side by side ip; ie ee Carmyertus
One testis dorsal of the other, Wein e in fmid- line o re Fischoederitus

Genus Gastrothylax, Poirier, 1883.

Fischoeder (1903) placed all the known Paramphistomidae with a ventral
pouch in the genus Gastrothylax. He divided the species in this genus
into five groups, using the shape of the ventral pouch on cross-section as
the distinguishing feature. His list is as follows :—

Genus Gastrothylax.

(a) Transverse section of pouch triangular with apex dorsally directed
and apical angle undivided.
I. G. crumensfer (Crepl.).
2. G. compressus, Brandes.

170

Fic. 13. Gastrothylax crumenifer. ‘Transverse sections through the middle of ten specimens

showing variation in shape of ventral pouch.
v.s.—vesicula seminalis. x 9.

7,—intestine ; “¢.—uterus ; v.p.—ventral pouch ;

171

(b) Transverse section of pouch triangular with apex dorsally directed
and apical angle bifurcated.
3. G. gregarius, Looss.

(d) Transverse section of pouch triangular with apex ventrally
directed.
. synethes, Fischdr.
. elongaius, Poirier.

Sys
AND

. cobbold1, Poirier.
8. G. mancupatus, Fischdr.

“NI

(ec) Transverse section of pouch triangular with apex ventrally
directed. The two basal angles bifurcated.
9. G. minutus, Fischdr.

The writer has carefully investigated the shape of the ventral pouch in
cross section in all the species he has had at his disposal. Fig. 13 illustrates
transverse sections at the mid point of ten specimens of G. crumenzfer.
All the specimens agreed in other anatomical details and all came from the
same bottle. Fig. 14 represents similar sections of six specimens of
G. spatiosus. |

On examining the drawings it will be seen that fig. 13, D, corresponds
with Fischoeder’s group (a), fig. 13, H and K, correspond with his group (8),
and fig. 13, F, is so nearly circular as to correspond with his group (e).
Fig. 13, A, B,C, E and J, do not agree with any of Fischoeder’s five groups.
Therefore, in a series of ten specimens of a single species three of Fischoeder’s
five groups are represented, and the others cannot be classified by this
method. If fig. 13, D, H and K, are further examined, it will be noted
that they are all triangular with the apex dorsal, whereas in fig. 14, B and D,
which approach the triangular shape, the apex is ventral. This at first
seems to indicate that a division between the species might be made on
the fact that when the pouch is triangular, the apex is ventral in some
species and dorsalin others. But fig. 14, A, upsets this view, because here
is a worm considerably distorted in fixation with the dorsal surface drawn
to the right and the ventral surface displaced to the left. The ventral
pouch in this case is a narrow triangle with the base to the right. If this
worm could be straightened up, one would have a specimen with the base

172

of the ventral pouch looking towards the ventral surface, unlike the other
examples of the same species in which the base of the pouch, when triangular,
is dorsally directed. From this it will be seen that the cross section of the
ventral pouch may assume almost any shape and cannot therefore be
regarded as of any use’in diagnosis. :

V.p.

Fic. 14. Carmyerius spatiosus. ‘Transverse sections through the middle of six specimens
‘ showing variations in shape of ventral pouch, 1.—intestine ; ut.—uterus; v.p.—ventral pouch ;
v.s.—vesicula seminalis. xX 9g.

The shape of the ventral pouch when seen from the side also varies
considerably, as reference to figs. 15, 16, and 17 will readily show. For the
most part the changes in shape of the pouch when viewed in this position
are caused by outpocketings from the posterior end along the dorsal and
ventral surfaces of the testes, but in a few cases a narrow prolongation may
also occur from the dorso-anterior part of the pouch (see fig. 17, C). In
addition to differing as a whole, the pouch also exhibits varying form in

173

Fic. 15. Gastrothylax crumenifer. Sagittal sections near the mid-line of three specimens
to show the shape of the posterior end of the ventral pouch. ov.—ovary ; p.s.—posterior sucker ;
t.—testis ; ut—uterus; v.g.—vitelline gland; v.p.—véntral pouch. x 9.

Fic. 16. Gastrothylax crumenifer. ‘Three sagittal sections from a single specimen at different
levels, showing change in shape of posterior end of pouch in one worm. ¢x.b.—excretory bladder ;
i.—intestine ; ov.—ovary ; p.s.—posterior sucker ; ut.—uterus; v-p.—ventral pouch. X 9.

174

different sections of the same worm (fig. 16). It will be noted later that
Stiles and Goldberger use the presence of a ventral prolongation of the
pouch beneath the testes as a specific character in at least one instance.

Fic. 17. Carmyertius spatiosus. . Sagittal sections of five specimens near the mid-line showing
variation in shape of posterior end of pouch. 4, B, D, and E—Gravid worms. C—Immature
worm. 4.s.—anterior sucker ; ex.b- excretory bladder. ; i.—intestine ; 0.—oesophagus ;
ov.— ovary; p.p.—pars prostatica; p.s.—posterior sucker; ¢.—testis; w/—uterus; v.g.—vitelline
gland; v.p.—ventral pouch ;_ v.s.—vesicula seminalis. X 9g.

The above demonstration of the presence or absence of this prolongation
in a single species (fig. 17) is held to prove that this character is of no value
for specific diagnosis.

Genus Gastrothylax (Poirier, 1883), s. str., Stiles and Goldberger, 1910.
Definition.—Gastrothylacidae : with the uterus crossing from one side
of the body to the other about its middle.

Type species Gastrothylax crumenitfer Wha 1847).
Only one species recorded.

175
Gastrothylax crumenifer (Creplin, 1847).
Synonymy :—Gastrothylax compressus, Brandes, 1898.

Stiles and Goldberger (1910) revised Fischoeder’s classification and
‘restricted the genus Gastrothylax to include only those species in which the
uterus crosses from one side of the worm to the other about midway between
the anterior and posterior extremities. The result of this restriction is
that the genus now contains only two species, viz., G. crumentfer and
G. compressus. The writer has not been able to consult the original
descriptions of these species, but Fischoeder (1903) describes them very
fully and use has been made of this work in the present discussion.

The points of difference between these two worms are summarised
from Fischoeder (1903) in Table IX.

‘PABLEW IX.

Points of difference between G. compressus and G. crumentfer.

G. compressus G. crumenifer
Length of oesophagus ...| 4oogs to 500/4 1'2 mm. to I°5 mm.
Gut caeca ae ...| Almost straight. Wavy.
End 300/4 to soot anterior to testes. | End behind anterior border of testes.
mterus ... i ...| Not so convoluted. More convoluted.
Eggs oat ree ...| Not so numerous. More numerous.
115fe to 12544 by 6o4e to 651. 12544 to 1354e by 6544 to Jo".

In the present instance the material available for examination consisted
of two bottles containing several hundred specimens. One collection came
from a bullock in India and the other from a bullock in Hong Kong.
Examination of a large number of these worms has shown that like all other
Amphistomata, they are liable to considerable variation. In our collections
there are many worms which could be identified as one or other of the
above species, and there are just as many which would fit either species
just as well. It is therefore considered that these two species have been
separated from one another on points of individual variation, and that
the differences are so variable in degree that in all probability G. compressus
is merely a synonym of G. crumenifer.

176
Genus Fischoederius, Stiles and Goldberger, Igto.

Definition.—Gastrothylacidae : with the uterus in the centre of the body
for its whole length ; one testis dorsal of the other.

Type species Fischoederius elongatus (Poirier, 1883), Stiles and.
Goldberger, IgIo. |

Key To SPECIES

Gut caeca extend at least to testes ... ; I. cobboldi
Gut caeca never extend further than just beyond Ea of body _ F, elongatus

Fischoederius elongatus (Poirier, 1883), Stiles and Goldberger, 1910. |
SYNONYMY :—

Fischoederius fischoederi, Stiles and Goldberger, 1910.

Fischoederius stamensis, Stiles and Goldberger, 1910.

Fischoederius ceylonensis, Stiles and Goldberger, 1910.

First found in the stomach of Palonia frontalis in Java.

The material available to the writer consisted of about 25 specimens
obtained from a bullock in Hong “Kong.

EKight specimens were cleared in carbolic acid and their examination
showed that the worm closely agrees with Fischoeder’s (1903) description,
with the proviso that this author does not take cognisance of individual
variations. For instance, Fischoeder states that the caeca end within
the beginning of the hinder half of the body, but in the eight specimens
examined, this was found to be the case only in three of them ; in one the
caeca ended exactly at the junction of the anterior and posterior halves of
the body, and in the remaining four specimens they ended at varying
distances up to as much as I mm. in front of it. In three cases the two
caeca were of unequal length ; in the most marked instance the gut on one
side was I mm. longer than on the other. In some of the worms examined,
the caeca were as convoluted as Fischoeder shows in his fig. 59, but in
others they were nearly straight, and furthermore some caeca presented
short dilated portions in various parts of their course, while in others they
were of uniform diameter throughout.

Fischoeder also says that the testes lie one above the other near the
mid-line, the ventral one being slightly to the right*of the mid-line and
slightly posterior to the dorsal, which is rather to the left of the mid-line.
This was true of some of the specimens examined, but in others one testis

appeared to lie directly above the other, and both of them were exactly
in the mid-line. |

eb
Fischoederius fischoedert, Stiles and Goldberger, 1910.

This species is stated to have been made from a single specimen found
in a bottle containing a number of worms which had been determined by
Fischoeder as G. elongatus (Ff. elongatus). They distinguished it from the
latter on the ground that the gut caeca were very slightly longer, and that
the ovary and shell gland lie between the testes in F. fischoederi, which
they state is not the case in F. elongatus. With regard to the position of
the ovary in F’ elongatus, Fischoeder (1903) states :

‘Der fast kuglige Keimstock (0°3 bis 0°35 mm. im Durchmesser) liegt dorsal von
der Schalendriise, etwas seitlich von der Medianlinie dicht hinter und etwas median
von dem meist nach hinten herabhangenden dorsalen Ende des vordern Hodens
(Text fig. L).’

It will be noted that he makes no reference to the other testis, but
Text fig. L, to which he refers, shows the ovary and shell gland lying
between the two testes, but a little further back than Stiles and Goldberger
show them in fig. 2. As it is recognised that the ovary varies slightly in
position in most species, this difference cannot be considered of specific
importance, especially as it relates to an isolated specimen taken from
a collection of F. elongatus which had already been determined by
Fischoeder. It is therefore considered that F. fischoederi is a synonym of —
F. elongatus.

Fischoederius siamensis and F. ceylonensis, the other two new species
made by Stiles and Goldberger, are separated from F. elongatus because
they do not exhibit the prominent bulging round the genital pore which
these authors state is present in [’. elongatus. It is true Fischoeder (1903)
in his description of F. elongatus mentions this bulging, but from the expezi-
ence of the writer, it is concluded that the bulging is a variable character
and is similar to that which may be found in any species of the group
Amphistomata and is accordingly of no specific value. It is therefore
considered that F’. stamensts is synonymous with F. elongatus.

F, ceylonensis was established as a new species on a single specimen
taken from a bottle of G. synethes. Stiles and Goldberger distinguish
between F. ceylonensis and F’. siamensis by two points ; firstly, the testes
in F. ceylonensis are directly one above the other and in F’. stamensis they
are slightly diagonal ; and secondly, the ventral pouch in IL. ceylonensis
extends ventral to the testes, whereas in Ff. stamensis this extension of the
ventral pouch does not occur. The individual variations which may be
found in the ventral pouch have, however, already been fully discussed,

178

and it is considered that the differences between F. ceylonensis and
F. siamensis are not of specific value, more especially as the diagnosis of
one of them rests on a single specimen.

Fischoederius cobboldi (Poirier, 1883), Stiles and Goldberger, 1910.

First found in the stomach of Palonia frontalis in Java.

No material of this species was available to the writer.

From Fischoeder’s description, the distinguishing feature between
F. cobboldi and F. elongatus is the difference in position of the termination
of the caeca. In F. cobboldi they end opposite the base of the sucker, that
is posterior to the testes and near the hinder end of the worm, and in
F’, elongatus they end about the middle of the worm. Although it has been
shown that the point of termination of the caeca is subject to variations in
practically all species of the group, in the present instance the differences
between the two species under discussion are so marked that it is unlikely
that they can be explained in this manner. It is accordingly considered
that F. cobboldi and F. elongatus can be distinguished by the difference
in length of the caeca.

Genus Carmyerius, Stiles and Goldberger, Igro.
Synonymy :—Wellmanius, Stiles and Goldberger, 1910.

Definition.—Gastrothylacidae : with the uterus in the centre of the worm
for its whole length ; testes side by side.
Type species Carmyerius gregarius (Looss, 1896), Stiles and Goldberger,

IgIo.
Key To SPECIES
I. Genital pore lies outside ventral pouch ee ve mS C. ee
Genital pore lies within ventral pouch 2

2. Excretory canal and Laurer’s canal unite before reaching the surface C. wenyoni
Excretory canal and Laurer’s canal do not unite

3. Cross section of ventral pouch shows five angles 1 mh C. cruciformis
Cross section of ventral pouch does not show five angles... AM ae
4. Gut caeca extend to testes aRt 8% bo C. spatiosus
Gut caeca do not extend beyond middle of worm... ~ C. gregarius

Stiles and Goldberger made this genus to include all the species of
Poirier’s Gastrothylax in which the uterus occupies a central position for
its whole length, the testes lie side by side, and the vas deferens is without
a straight portion at its commencement. This included the following five

{79

species: C. synethes (Fischoeder, 1gor), C. gregarius (Looss, 1896),
C. spatiosus (Brandes, 1898), C. mancupatus (Fischoeder, tgor), and
C. minutus (Fischoeder, tgo1). They divide the above five species into
two groups, because the first two worms are said to have a genital atrium
with a large ventral chamber and the last three a genital atrium without a
ventral chamber. These characters have been fully discussed already
and it has been shown that presence or absence of chambers in the atrium,
or even the existence of the atrium itself, are purely matters of chance,
so that it seems reasonable to assume that this point is of no value in the
present instance. Differences in shape of the ventral pouch are also
mentioned in the definitions of the various species. The unreliability of
this character has already been dealt with.

Carmyerius spatiosus (Brandes, 1898).
SYNONYMY :—
Carmyerius synethes, Fischoeder, 1got.
Carmyerius minutus, Fischoeder, 1901. -
Carmyerius mancupatus, Fischoeder, Igo.
Gastrothylax bubalis, Innes, 1912.
Wellmanius wellmant, Stiles and Goldberger, 1910.

First found in the stomach of Bos taurus in Arabia.

Material available :—Two collections from Ngoa, N.E. Rhodesia ;
the host in one case was a roan (Hippotragus equinus) and in the other a
reedbuck (Cervicapra sp.).

As the four species C. spatiosus, C. synethes, C. minutus and C. mancu-
patus only differ in minute points apart from those already dealt with,
they will be discussed together.

According to Stiles and Goldberger’s key, the only point by which these
four species may be distinguished are minute differences in the gut caeca.
They write that in C. synethes the caeca are ‘ corkscrew-like, rather narrow
and long’; in C. spatiosus they are ‘ straight, narrow and rather long’ ;
in C. mancupatus they are ‘rather sinuous, narrow, and long’; and in
C. minutum they are ‘ swollen in their caudal half and are rather long.’
In all four cases the caeca are said to end in the ‘ fourth zone.’ Twenty
specimens were cleared in carbolic acid and it was found that the degree of
convolution of the caeca was so variable as to be of no use for diagnosis.
With regard to differences in diameter of various parts of the caeca, they
were found swollen or contracted in any part of their course or of uniform

180

diameter throughout. Fig. 18, A and B are drawings of the two caeca in
a single worm showing alternate contractions’and dilatations fie TOC
a drawing from another specimen showing the gut of uniform diameter from
end to end. It is therefore clear that dilatation of a special part of the
caeca is of no diagnostic value.

vg.

Fic. 18. Carmyerius spatiosus. Sagittal section to show variation in calibre of gut.
A and B—Showing the two sides of the same worm. C—Section of another worm. a.s.—anterlor
sucker ; 1.—intestine ; ov.—ovary ; p.s.—posterior sucker ; 4.—testes ; ut.—uterus ; v.g.—vitelline
gland ; v.p.—ventral pouch ;_ v.s.—vesicula seminalis. x 9.

Fischoeder (1903) mentions one or two other small points of difference,
viz., the genital pore lies further posteriorly in C. synethes than in any other
species, being about 1 mm. behind the entrance to the ventral pouch ; but
as he gives 0-8 mm.-o-g mm. and 0-7 mm.-o-8 mm. for the same dimension
in C. spatiosus and C. mancupatus respectively, it cannot be considered
justifiable to regard this of specific value, especially as he gives the limits
of variation in length of the worms as being between 6 mm.-12 mm. for
the three species.

- 2Pe

181

Fischoeder states that C. minutus is 4 mm.-5 mm. in length; then he
remarks that some specimens are longer and thinner than this, but gives no
other measurements. This point cannot be regarded of any value when he
says that C. spatiosus, C. synethes and C. mancupatus are 9 mm.-I2 mm.,
7 mm.-II mm., and 6 mm.-11 mm. in length respectively. Another point
of difference claimed between C. minutus and the other three species, is
that the ovary and shell gland are said to be more strongly developed in the
former. But difference in size of the genital organs is so dependent on the
age of an individual that it cannot be regarded as a satisfactory diagnostic
character.

From the above observations it is concluded that Fischoeder’s three
species, C. synethes, C. mancupatus and C. minutus are synonyms of
C. spatiosus (Brandes).

Carmyerius wenyont (Leiper, 1908).

Found in the stomach of Cobus maria at Taufikia, White Nile.

It is not included in Stiles and Goldberger’s (1910) classification.
It is similar in most respects to C. spatiosus, but there is one point by
which it may be clearly distinguished, and that is that the excretory canal
and Laurer’s canal unite before reaching the surface of the worm and
open by acommon pore. This character is very remarkable as it has not
been observed in any other species.

2
Carmyertus cruciformis (Leiper, IgIo).

The material available for study consisted of a single collection ot
over one hundred specimens from a hippopotamus killed in Lake Victoria,
Nyanza.

This worm was first described by Leiper who states that he only had
immature material. Leiper gives the length of this worm as from 0.5 mm.
too’8mm.; this is apparently a mistake for 5mm. to 8 mm. because he
gives the dorso-ventral diameter of the posterior sucker as 0°9 mm.
with a muscle wall of 18mm. in thickness. Some of the writer’s
specimens contained eggs in the uterus and these worms measured 5 mm.
to 7mm. in length with a maximum diameter of about 1°5 mm.; they
agree in general anatomical details with Leiper’s description. The uterus

182

was observed to pursue a slightly wavy course along the centre of the
dorsal surface; the most heavily gravid specimen only contained about
thirty eggs disposed in a single chain and not closely packed into the
uterus in the usual manner. Eggs removed from one specimen measured
about 135-140 in length by 80-84 in breadth; on account of the small
size of the worm they appeared relatively large. Leiper named this
worm ‘cruciformis’ because he says the ventral pouch always shows five
angles in cross section no matter how contracted the worm may be, and
this frequently appears as a cross. The writer cut eight specimens
transversely, and although the pouch of three of them exhibited five
angles none were at all like a cross and the other five specimens
exhibited just as extensive variation as do all Gastrothylacidae in respect
of the shape of the pouch.

This worm is essentially the same as C. spatiosus in details of
anatomy, but is easily distinguished from this species by its minute size
even in the gravid state.

Gastrothylax bubalis, Innes, 1912.

This worm was recorded by Innes (1912) as a new species. It was
found in the stomach of the Hartebeest in Rhodesia.

The general anatomy of the worm is apparently in no way different
from C. spatiosus ; to quote from Innes the identification of the species is
based on the following characters :—

‘The excretory vesicle of this species is unique both in position and outline.
It is situated immediately in front of the posterior sucker and behind the shell gland,
while in other species it has a more or less lateral position. ‘The great irregularity
of its outline is striking.’

No authority is given for the statement that the excretory bladder lies
in a more or less lateral position in other species, and Fischoeder (1903)
states that in G. spatiosus :

‘Die sehr grosse Excretionsblase liegt zwischen der Bauchtasche und dem
Saugnapfe einerseits und den beiden Hoden andrerseits (fig. 52c. u. 54).’

And in all other species of the genus.that he deals with he describes it
in a similar position, only alluding to slight alterations due to change of
shape, and his figures all show it in the same position as it is seen, Innes’
fig. 7. It therefore seems clear that this difference claimed by Innes does
not exist. With regard to his other point, viz., the irregularity of the
excretory bladder, it may be observed that irregularity of outline is

183

characteristic of a partly distended bladder in any species, and is obviously
of no specific value. From this it is concluded that the species G. bubalis
is only a synonym of C. spatiosus.

Carmyerius gregarius (Looss, 1896), Stiles and Goldberger, rgro.

First found in the stomach of Bos bubalis in Egypt.

Material available.—A single collection of about 25 specimens from
a buffalo.

This species is easily distinguished from the other species of the genus,
as the gut caeca are very short. Of ten specimens cleared in carbolic acid
only one was found in which the caeca reached the middle of the worm ;
in all the others they ended at varying distances anterior to this point.
For the most part the caeca were of uniform diameter throughout, but in
one specimen they exhibited a swelling about their middle and in another
there was a club-shaped swelling at their extremities. Although variation
in the length of the caeca has been found unreliable for specific diagnosis
when the range is slight, they are, however, so much shorter in C. gregarius
than in C. spatiosus that this difference is considered of specific value in the
present instance.

Carmyertus exoporus, N.Sp.

Found in the stomach of a Tvagelaphus speket in Nyasaland.
The material available was a single collection of over three hundred
specimens.

EXTERNAL ANATOMY

All stages of growth were represented in the collection, from worms no
bigger than a raspberry seed up to gravid worms measuring Over II mm. in
length. But even in gravid worms the shape and size was subject to great
variation (see Plate VII, A). It will be noted in this plate that in some
cases the worms are fully extended (9) and in others they are in a state of
contraction (7). In view of the great variation in shape that exists, it is not
considered worth while giving a detailed description, but as a rough
indication of what the size of gravid worms is, it may be stated that they
varied from I1-5 mm. in length by 2-6 mm. in breadth to 5 mm. in length
by 3°8 mm. in breadth. In all cases the worms were practically circular
on cross section. As the worms are not curved ventrally, the opening of
the anterior sucker lies at the extreme anterior end and looks directly

184

forwards. The opening of the ventral pouch lies in the mid-line of the
ventral surface close behind and below the oral opening, and between the
two is the opening of the genital pore, which is thus outside the ventral
pouch. This character is sufficient to distinguish the present species from
any other member of the genus Carmyerius, as in all the hitherto described
species the genital pore opens within the ventral pouch. The posterior
sucker, as a rule, looks directly backwards, but in a few cases it is slightly
tilted ventrally.

INTERNAL ANATOMY

The general arrangement of the organs of this species is very similar
to G. spatiosus and on that account only the briefest description is necessary.

Muscular system. This system exhibits no special characters.

Nervous system. This system was not investigated.

Anterior sucker. The anterior sucker is of the typical globular shape,
oval in section.

Intestines. The oesophagus arises from the posterior end of the
anterior sucker and after pursuing a dorsally curving course of about 300p,
it divides into the two gut caeca. These two canals pursue a wavy course
along each side of the worm and terminate about the level of the testes.

Posterior sucker. The posterior sucker is fairly thick-walled and looks
directly backwards or slightly towards the ventral surface (figs. 20, 21 A,
and 25). |

Excretory system. The excretory bladder is of the usual type and its
degree of convolution varies with its state of distension. The excretory
canal runs dorso-posteriorly and opens in the mid-line of the dorsal surface
above the anterior border of the posterior sucker (fig. 20).

Gemtalia. Testes. The testes are large lobed organs lying one on each
side of the mid-line near the hinder end of the worm (fig, 21 A).

Vas deferens. This duct is composed of the usual three portions, the
vesicula seminalis, the pars musculosa, and the pars prostatica. The first
two portions are much convoluted and occupy the anterior third of the
centre of the dorsal part of the worm. The pars prostatica is relatively
long and straight, and runs directly forwards to enter the genital papilla
(figs. 19 and 21, B).

Genital pore. The genital pore is surrounded by a muscular thickening
not marked off from the parenchyma and it opens in the middle of the worm
between the oral opening dorsally and the opening of the ventral pouch

185

ventrally (figs. 19 and 22). It is of the usual type in structure and the
appearance shown in the figures is not the only one met with, as the genital
papilla is capable of complete retraction or extrusion as in other species.
In the figures it is in the intermediate condition.

Ovary and shell gland. These two structures lie between the testes on
one hand, the base of the ventral pouch in front and the posterior sucker
on the other (figs. 20 and 21 A).

Laurer’s canal. Laurer’s canal runs almost directly dorsally from the
shell gland and opens in the mid-line well in front of the excretory pore
(fig. 20).

Vitellaria. For the most part these glands lie in the ventral portion
of the worm, but they extend for varying distances on each side towards
the dorsal surface. As a rule, they extend from the posterior end of the
anterior sucker in front, to opposite the testes behind, but in a few cases
follicles were found close to the posterior end of the worm, and surrounding
the opening of the posterior sucker (figs. 19, 20, 21, A, and 25).

Fic. 19. Carmyerius exoporus, n.sp. Sagittal section. a.s.—anterior sucker; g.p.—genital
pore; 0.—oesophagus ; p.m.—pars musculosa; p.p.—pars prostatica; ¢.— testis; wut.—uterus ;
v.g.—vitelline gland ; v.p.—ventral pouch. X18.

Uterus. The uterus pursues a wavy course along the centre of the
dorsal surface ; anteriorly it terminates by running ventral to the pars
prostatica and uniting in the genital papilla with the male duct (figs. 21 B
and 22).

Eggs. These are oval, operculated and measure about 115m to 130y in
length by 60u to 68 in breadth, but they were only taken from one
specimen, so these dimensions must be regarded as only approximate.

Ventral pouch. The ventral pouch is somewhat characteristic in

186

Fic. 20. Carmyerius exoporus, n.sp. Sagittal section of posterior end of worm.
ex.b.—excretory bladder; ex.p.—excretory pore; L.c.—Laurer’s canal; ov.—ovary; p.s.—posterior
sucker ; sb.g.—shell gland ; ut.—uterus; v.g.—vitelline gland; v.p.—ventral pouch. x 16.

Fic. 21. Carmyerius exoporus. Coronal sections. d—About middle of worm. B—Near
dorsal surface. a.s.—anterior sucker; i.—intestine; ov.—ovary ; p.s.—posterior sucker ;
sh.g—shell gland; t.—testes; wut—uterus; v.g.—vitelline gland; v.p.—ventral pouch;
v.S.—vesicula seminalis. x 8.

187

some cases, in that on cross section about its middle it often shows six points
or angles; but it is subject to so many variations that this cannot be
regarded as a diagnostic character. These variations are well shown
in figs. 23, 24 and 25.

DIAGNOSIS

As this worm is easily distinguished by the fact that the genital pore
opens outside the ventral pouch, the name Carmyerius exoporus, n.sp., is
_ suggested.

Genus Wellmanius, Stiles and Goldberger, Igto.

This genus was established by Stiles and Goldberger to include a new
species Wellmanius wellmani which was obtained from the stomach of
a reed bok (Cervicapra bohor) at Benguella, West Africa. The sole
distinguishing character between Wellmanius and Carmyerius is that in the
former the first part of the vesicula seminalis is straight, and in the latter
it is coiled from the commencement. This seems a very small point on
which to establish a genus, especially when it is found that the beginning of
the vesicula seminalis cannot be made out with accuracy in most gravid
specimens whatever the species may be, because it is overlaid by the uterus.
W. wellmani appears to the writer to be synonymous with C. spatiosus.

Family GASTRODISCIDAE, Stiles and Goldberger, 1910.

Definition. A mphistomata : body usually flattened and divided into
anterior and ot SSE portions.
Key To GENERA

1. Anterior portion large and flat, and posterior portion smaller and spherical

Homalogaster

Anterior portion small and conical, posterior portion large and flat ers 2,

2. Genital pore on anterior portion, ventral surface of posterior portion not
covered with papillae ... 40 ... Gastrodiscoides
Genital pore on posterior portion, ventral surface of posterior portion covered
with papillae ... Ae ae ae ee pe — Gastrodiscus

Genus Gastrodiscus, Leuckart, 1877.
Defimition.—Gastrodiscidae: anterior portion small and _ conical,
posterior portion large and flat; genital pore on posterior portion,
ventral surface of posterior portion covered with papillae.
Type species Gastrodiscus aegyptiacus (Cobbold, 1877), Looss, 1896.
Key To SPECIES

Genital pore less than I mm. from anterior border of posterior portion G. aegyptiacus
Genital pore over I mm. from anterior border of posterior portion G. secundus

showing variation in shape of ventral pouch.

x 9:

rGa22e

Fic. 23.

Carmyerius exoporus, n.sp.
a.s.—anterior sucker; g.p.—genital pore ;

pouch. X 30.

Hi) i
jail al AN

aii

r Li Ani) 1 i i} Dy.
AVANT as Pe h i
\ ys a TAN AY
|
MAAN Wt if Vii
i M iM !
Ze nie ‘ i inh )
f Ey la is ys Me
* oie
NY a

Sagittal section on enlarged scale to show genital pore.
p.p.—pars prostatica; wt.—uterus; v.p.—ventral

A.
ut.

Carmyertus exoporus, n.sp. ‘Transverse section of eight specimens near the middle
1.—intestine ; ut.—uterus ; v.p.—ventra! pouch. °

Fic. 24. Carmyerius exoporus, n.sp. Seven transverse sections of a single worm at different
levels showing alteration in shape of ventral pouch. a.s.—anterior sucker; i.—intestine ; p.p.—pars
prostatica. .—testes; mt.—uterus;]v.e.—vas efferens; v.p.—ventral pouch; v.s.—vesicula
seminalis. x 12.

.
*.*
.

_ _ Fic. 25. Carmyerius exoporus, n.sp. Sagittal sections of four specimens to show differences
in shape of ventral pouch. A, C,and D—Gravid worms. B—Immature worm. ex.b.—excretory
bladder ; p.s.—posterior sucker; ¢.—testis; wt.—uterus; v.g.—vitelline gland; v.p.—ventral
pouch; .s,—vesicula seminalis. x 9.

Igo
Gastrodiscus aegyptiacus (Cobbold, 1877), Looss, 1896.

SyNoNnyMy :—Gastrodiscus minor, Leiper, 1913.

First found in the horse.
The material available consisted of the following collections :—

1. Over one hundred specimens from the large intestine of a pony at Ilorin,
Northern Nigeria.
2. Over one hundred specimens from the large intestine of a zebra (Equus sp.) in
Rhodesia. ;
3. Over one hundred specimens in poor condition passed by a horse in Northern
Nigeria.
About fifty specimens from a horse at Nairobi, Kenya Colony.
Six specimens from a mule at Nairobi.
Three collections from wart-hogs (Phacochoerus sp.) in Ngoa, North-East
Rhodesia.

Sb i Meese

As the first four collections consisted mainly of relatively large worms
(about 15 mm. in length), and the last four mainly of smaller worms
(about 9-12 mm.), it was at first thought that two distinct species were
present, but it was found on detailed examination that no differences in
anatomy could be discovered, and, as in both types gravid worms were
found, it was assumed that the difference is nothing more than a size
variation of the one species.

Looss (1896) in his description of G. aegyptiacus states that the testes
are arranged diagonally, with the anterior of the two on the right side
and the posterior on the left ; the ovary was on the left side behind the
posterior testis. Among specimens examined in the present instance,
it was found that the relative position of the testes varied and that some-
times the left testis was anterior. It was also noted that the ovary was
invariably on the same side as the anterior testis.

Vitellaria. The vitelline glands are described by Looss as composed
of two groups of follicles arranged near the ventral surface, and along the
outer side of the gut caeca in the posterior disc-like portion of the worm,
and confined to these two areas, except that in some cases they may spread

inwards so as nearly to meet on the dorsal side of the hinder ends of the gut |

caeca (this arrangement is shown by the heavy dots in fig. 26). Among the
present collections of worms, the restricted type of distribution of the
vitellaria was noted in two collections only, viz., that from the pony, Ilorin,
and from the zebra, Rhodesia. In all the other collections the vitellaria
were found to be much more extensive, though somewhat variable in
distribution. In fullest development the vitellaria not only extended right

Ig!

across the dorsal surface of the posterior disc-like part for its whole length,
but also into the cephalic portion, where at times they were very thickly
massed and completely encircled this part of the worm (fig. 26). The
number of gland groups varied very much in different individuals, especially
in the cephalic part of the worm. In a few cases the vitellaria in the
caudal part of the worm were almost completely confined to the inter-
caecal field on the dorsal aspect. In these worms the vitellaria extended

Fic. 26. Gastrodiscus aegyptiacus. Dorsal surface uppermost. a.s.—anterior sucker ;
0.—oesophagus; o0.d.—oral diverticulum; g.p.—genital pore; 17.—intestine; 0v.—ovary ;
p.s-—posterior sucker ; #1.—anterior testis ; t2,.— posterior testis; ut.—uterus ; v.g.—vitelline

gland. x 6,

as a single broad band from the anterior end to the posterior end of the
worm. It might be considered that these differences are of specific value,
but in view of the great variation found in the distribution of the vitellaria
in all other species of Amphistomata that the writer has examined, one is
compelled to consider them of no specific value.

With the object of determining the exact position of the genital pore
in relation to the anterior edge of the caudal portion of the worm, nineteen

192

specimens from the various collections were examined (see Table X).
It was not possible to examine more specimens in this particular point,
because the edge of the caudal part is usually incurved, so that it requires
considerable pressure, and hence distortion, to flatten it out sufficiently
to measure the distance between the genital pore and this edge.

TaBLe X

Distance between genital pore and anterior edge of discal portion of worm

Wart-Hog ... a es * Small 572 9°9 mm. Die
6254 I1‘O mm. 1 oii

416u g'I mm. 13203

416u 8°g mm. Is 2

Wart-Hog ... ¥e a Small 468 10°O mm. Li ote
731 I1°4 mm. I: 14°6

572u 10°4 mm. 1.3 18%

625 98 mm. ere si:
400M 8°3 mm. I $2075

Mule, Nairobi ... be Small 468u 8°3 mm. I 186
572 9°6 mm. I: 168

Zebra AM 5% A Large 731M 13°3 mm. Lia
729 13°0 mm. L213

Pony, Ilorin sae ee Large 760M 13°5 mm. Bg ie
Horse, Nairobi... es Large 625 11°3 mm. Lyioee
833 12°5 mm. 1 iit

677 I2°0 mm. t i797

639h 11°25 mm. 1 178

625 13°75 mm. i‘ Pz20

It will be noted from the above table that the distance between the
genital pore and the anterior edge of the caudal part of the worm varies
considerably. Another point which this series of measurements brought
out is that although on the whole the two types are fairly distinct, relatively
small worms may be found in bottles in which the predominating number
are large, and in bottles that contain nearly all small worms a few
relatively large ones occur, and these exceptional specimens tend to
unite the two types into a complete whole. From these observations it is

concluded that the species G. aegyptiacus is subject to considerable size
variation, and that the distance of the genital pore, although further from
the edge of the caudal part of the worm in large examples than it is in small
ones, is variable.

Gastrodiscus minor, Leiper, 1913.

This species is recorded by Leiper (1913) as new, the host being the
pig in Uganda. All Leiper gives in the way of description is the following
passage : |

‘This small fluke resembles closely the African G. aegyptiacus (vel sonsinoi),
which is so frequently met with in horses in Egypt and in West Africa: it differs,

however, in a number of respects, particularly in the nearness of the genital pore
to the edge of the ventral disc-like expansion.’

From this it is rather difficult to deduce in what the differences between
G. minor and G. aegyptiacus consist, especially if reference is made to
Table X. G. minor is therefore regarded as a synonym of G. aegyptiacus.

Gastrodiscus secundus, Looss, 1907.

The material available for study consisted of a single collection of
about twenty specimens. It was from the same collection that Looss
obtained the material he used in his description of the species.

Examination of this material did not reveal any differences from the
original description by Looss (1907), except that the position of the testes
and ovary varied in the same manner as has been described in
G. aegyptiacus. The worm can at once be distinguished from G. aegyptiacus
by the position of the genital pore. Fig. 27 shows the essential anatomical
details.

Genus Gastrodiscoides, Leiper, 1913.

Definition.—Gastrodiscidae: anterior portion small and _ conical,
genital pore on anterior portion, no papillae on ventral surface of posterior
portion.

Type species Gastrodiscoides hominis (Lewis and McConnal, 1876),
Leiper, 1913.

Only one species described.

SYNONYMY :—

Amphistoma hominis, Lewis and McConnal, 1876.
Gastrodiscus hominis, Ward, 1903.

First found in the colon of man in Assam.

NN
SAN

fp
(i

Sa
=

WG.
Ny
q iy 1\\ \
yh

Lie
Zé,

Fic. 27. Gastrodiscus secundus. Dorsal surface uppermost. a.s.—anterior sucker;
g-p-—genital pore; 7.—intestine; 0.—oesophagus; 0.d.—oral diverticulum; 0v.— ovary ;
p-s-—posterior sucker; #1.—anterlor testis; %2.—posterior testis; m#t,—uterus ; v.g.—vitelline
gland. x 12.

Gastrodiscoides hominis (Lewis and McConnal, 1876), Leiper, 1913.

The material available for study consisted of :—

Two collections from Annam consisting of nine whole worms and six
specimens cut and mounted in serial section. This is the same material
as Stephens (1906) used in his description of the worm.

Leiper describes and figures (fig. 35) a prominent genital papilla with
the male and female ducts opening separately near its tip. Figs. 28 and 29
in the present paper were drawn from two sectioned specimens cut by
Stephens. In fig. 28, it will be noted that there is a deep atrium with no
sign of a papilla, the openings of the uterus and vas deferens being widely
separated from one another and lying at the deepest part of the atrium.
This represents a worm with the papilla completely retracted. In fig. 29
there is a bulbous papilla partly protruded, with a small atrium surrounding

Gastrodiscoides hominis.

Fic. 28.
Sagittal section of genital pore,with papilla

fully retracted. g.p.—genital pore;
p-p.—pars prostatica ; wt.—uterus X 30.

195

Fic. 29. Gastrodiscoides hominis.
Sagittal section of genital pore, with papilla
partly extruded. d.e.—ductus ejacula-
torius; d.b.—ductus hermaphroditicus ;
£.pp.—genital papilla; ut.—uterus. x 30.

its base, the male and female ducts uniting within the papilla and opening

at its tip by a common duct, in the usual way. These two drawings taken

in conjunction with Leiper’s figure indicate that the presence or absence of

a prominent genital papilla, or of a genital atrium, are purely matters of

chance, and are of no more diagnostic value in this instance than in any

other species of the group Amphistomata.

Genus Homalogaster, Poirier, 1883.

Definition.—Gastrodiscidae : anterior portion large and flat, posterior

portion smaller and spherical.

Type species Homalogaster poloniae, Poirier, 1883.

Only one species described.

Homalogaster poloniae, Poirier, 1883.

SYNONYMY :—

Homalogaster poirieri, Giard and Billet, 1892.

Homalogaster philippinensis, Stiles and Goldberger, 1910.

First found in stomach of Palonia frontalis, in Java.

The writer has not had the opportunity of examining any material of

_ this species, but Railliet, Henry, and Bauche (1914) discuss its synonymy,
and it is from their paper that it has been taken.

196
Brumptia, Travassos, 1921

Definition.—Amphistomata : with paired caudal appendages containing
most of the vitellaria, cirrus pouch and genital sucker present.
Type species Brumptia gigas.

Brumptia gigas (MacCallum, 1917) Travassos, 1921

SYNONYMY :—

Cladorchis gigas, MacCallum, 1917. |

This worm was found on two occasions in the stomach of a rhinoceros
at Ngoa, North-east Rhodesia.* Each collection consists of about
twenty-five specimens.

EXTERNAL ANATOMY

Size and shape. The worms were of slightly different size in the two
collections, those in one bottle being all about 15 mm. in length by 9 mm. in
breadth, and those in the other bottle about 12 mm. in length by 7 mm.
in breadth. Gravid worms were found in both collections, but as detailed
examination revealed no differences other than size, it is considered that
there was only one species. The worms consist of two portions distinctly
separated, an anterior conical portion and a posterior part consisting of two
crescentic flaps. The anterior part is conical in shape with a definite
ventral curve ; the ventral surface is almost flat from side to side, whilst
the dorsal surface is domed both laterally and antero-posteriorly.

The posterior sucker is slightly in front of the posterior extremity of
the body of the worm, and is situated entirely on the ventral surface, and
directed ventrally. About midway between the two suckers in the mid-line
of the ventral surface is the genital pore. Comparison of figs. 5 and 6,
Plate VIII, will show how the appearance of the genital pore varies with
retraction or extrusion of the genital papilla.

The most characteristic feature of the worm is the presence of two large

* The above description was written before I became aware that MacCallum (1917) had
described a worm which is apparently the same species. MacCallum obtained his material from the
African elephant (Loxodon africanus), he named it Cladorchis gigas, The Liverpool material seems to
be identical with MacCallum’s in all anatomical details, but the two collections are a little different
in size, MacCallum’s worms are 21 mm. in length, and ours are from 12 mm. to 15 mm. in length.
In view of the results obtained in other species this slight difference is not considered to be of
importance. Travassos (1921) created a new genus Brumptia to accommodate MacCallum’s species
C. gigas, leaving it in the sub-family Cladorchinae, but the two caudal flaps containing the vitelline
glands are so strikingly different from any other genus in this sub-family that I consider it preferable
to leave it as a genus of uncertain position.

197
crescentic caudal appendages. These arise from the _postero-lateral
borders, extend laterally as far forward as the anterior border
of the ventral sucker, and are separated from one another behind the
sucker by a deep notch. In full extension they measure about 5 mm. in
length, but as a rule, their borders are incurved towards the ventral
surface, so that they appear somewhat shorter and tend to overlap the

posterior sucker.

INTERNAL ANATOMY

On account of the thickness of these worms, very little could be
ascertained in whole specimens cleared in carbolic acid, so that the following
description is mainly based on a study of serial sections cut in sagittal,
coronal, and transverse planes.

Muscular system. The muscular system, as a whole, is very similar.
to other species, but in certain special structures it departs from the usual
type, and these differences will be dealt with under the appropriate organs.

Nervous system. This system was not investigated.

Excretory system. The excretory bladder is large when in a state of
distension and occupies the whole of the posterior part of the worm
between the posterior sucker and the dorsal surface. The excretory
canal in the specimens examined ran almost directly posteriorly to open
in the mid-line near the posterior end of the dorsal surface (fig. 30, B).

Anterior sucker. The anterior sucker is a thick walled muscular
structure surrounding the oral cavity ; about the junction of the middle
and posterior thirds there is an annular constriction at which point two
large muscular diverticula arise and run in a dorsal and slightly posterior
direction (figs, 31, A and 32, C).

Oesophagus and intestines. The oesophagus is of the usual type, its
muscular wall becoming slightly thicker as the posterior end is approached.
In the specimens examined, it curved at first ventrally, and then turning
abruptly towards the dorsal surface, divided into the gut caeca in front
of the cirrus pouch (fig. 30, B). The caeca pursue a wavy course along each
side of the worm and end in the dorsal part of the caudal flap (fig. 30, A).

Genitalia. Testes. These are large oval organs lying side by side in
the lateral fields somewhat nearer to the ventral than to the dorsal surface.
They lie in front of the posterior sucker, and the posterior part of the cirrus
pouch is between their anterior ends (figs. 30, A, 31, B, and 32, B). No
external lobing is visible, but in sections each testis is seen to be divided

198

up into numerous separate acini, the whole being surrounded by a loose
connective tissue capsule. Each testis is about 3-5 mm. in diameter.
Vasa efferentia. These arise from the antero-mesial aspect of each
testis, and suddenly dilate into broad thin-walled tubes, which, running
upwards and inwards over the posterior wall of the cirrus pouch, enter this
structure on its dorso-posterior aspect by two narrow tubes lying close to
each other. When they reach the inner aspect of the wall of the cirrus
pouch they unite to form the vas deferens (figs. 30, 31, C and 32, A).
Vas deferens. The vesicula seminalis is a dilated, thin-walled sac

4 ‘
> .
t t=
Lae;
ee e>,

“S

e
*

Fic. 30. Brumptia gigas, Sagittal sections. d—To one side of mid-line. B—In

mid-line. @.s.—anterior sucker; c.f.—caudal flap; c¢.p.—cirrus pouch; d.4,—ductus
hermaphroditicus ; ex.b—excretory bladder; ex.p.—excretory pore; g.p.—genital pore ;
g.s.—genital sucker; L.c.—Laurer’s canal; 0.—oesophagus; 0.d.—oral diverticulum ;

0v.—ovary 5 p.p.—pars prostatica ; p.s.—posterior sucker; ¢.—testis; ut.—uterus; v.¢.—vas
efferens ; v.s.—vesicula seminalis. x 6.

which runs along the dorsal wall of the cirrus pouch, being held in place
by some strands of connective tissue (figs. 30, B and 31, B). Near the
anterior end of the cirrus pouch on its dorsal surface, the vesicula seminalis
passes into the pars prostatica. The pars prostatica runs ventrally for
some distance close along the anterior wall of the cirrus pouch, and then
leaving the wall of the cirrus pouch turns sharply towards the dorsal
surface, curving posteriorly until it ends by uniting with the uterus near
the centre of the pouch (fig. 30, B). The pars prostatica is thickly

my)

surrounded by cells for its whole course and no pars musculosa could be
distinguished. The genital papilla appeared as a long muscular tube
running from about the centre of the cirrus pouch towards the ventral
surface (figs. 30, B and 31, B), but in all the specimens sectioned it wasina
state of retraction, and would probably appear quite different in sections of
a worm like that shown in Plate VIII, fig. 5, where it is obviously protruded.

Cirrus pouch. The cirrus pouch is a spherical organ about 4 mm. in
diameter. It lies near the centre of the worm slightly towards its anterior
end. Its wall is composed of loosely laminated muscular fibres. That
part of the cirrus pouch which is not occupied by sex ducts is filled with
loose areolar tissue.

et wy. ce J < !

ea} et

Fic. 31. Brumptia gigas. ‘Transverse sections. d—Near anterior end. B—Through
cirrus pouch. C—Through ovary. D—Through middle of posterior sucker. E—Through
excretory pore. c.f.—caudal flap; c.p.—cirrus pouch ; d.4.—ductus hermaphro-
diticus ; ex.b.—excretory bladder; ex.d.—excretory duct ; ex.p.—excretory pore; g.p.—genital
pore; g.s.—genital sucker; 7.—1intestine; L.c.—Laurer’s canal; 0.—oesophagus; 0.d.—oral
diverticulum ; /.s.—posterior sucker ; ¢.—testis ; ut—uterus; ut1.—ascending branch of uterus ;

utz.—descending brancb of uterus; v.e.—vas efferens; v.g.—vitelline gland; v.s.—vesicula
seminalis. x 4.

Genital pore. This is provided with a definite small sucker surrounding
its opening. This sucker is much more definitely marked off from the
subcuticular muscle than in the case of the genus Cotylophoron, as it is
composed of radially arranged fibres quite distinct from the subcuticular
muscle ; this is shown in figs. 31, B and 30, B, in both of which the genital
papilla is seen lying within the genital sucker and surrounded by a small
atrium, which would obviously disappear if the papilla were extruded.

200

Ovary. This lies towards the dorsal surface between the testes and
slightly to one side of the mid-line. It is a circular organ with no special
characters (figs. 30, B, 31,C, and 32, A). The shell gland lies on the mesial
aspect of the ovary (fig. 32, A).

Laurer’s canal, Laurer’s canal runs dorsally from the shell gland and,
curving posteriorly over the anterior end of the excretory bladder, it
opens in the mid-line above the middle of the bladder and far in front
of the excretory pore (fig. 30, B).

Fic. 32. Brumptia gigas. Coronal sections. _d—Towards dorsal surface. B—About
the middle. C—Near the ventral surface. c.p.—cirrus pouch; d.b.—ductus hermaphro-
diticus; g.p.—genital pore; g.s.—genital sucker; 1.—intestine; 0.—oesophagus; 0.d.—oral
diverticulum ; 0v.—ovary; p.p.—pars prostatica; p.s.—posterior sucker ; sh.g.—shell gland ;
t.—testis ; ut.—uterus; v.e.—vas efferens; v.g.—vitelline gland. x 4}.

Vitellaria. The vitelline glands consist of numerous collections of
follicles which lie nearly entirely within the two caudal appendages
(figs. 30, A, 31, C.D and E, 32, BandC). A few groups of follicles were seen
near the ventral surface of the worm in front of the posterior sucker.

201

Uterus. For the first part of its course the uterus shows no special
differences from the usual type. But after it reaches the anterior border
of the cirrus pouch on its dorsal aspect, it turns posteriorly and runs back,
still close to the dorsal wall of the cirrus pouch, and following the curve of
the posterior wall of this organ comes close to the ventral surface ; it then
turns sharply dorsally and enters the posterior wall of the cirrus pouch
about its middle. From this point, it runs anteriorly through the pouch to
unite near the centre with the end of the pars prostatica (fig. 30, B).

Eggs. The eggs removed from the uterus of one specimen were oval
and operculated and measured 112 to 116m in length by 76m to 7ou in
breadth, but it must be remembered that measurements of eggs taken
from the uterus of fixed worms are only very approximate.

SPECIES INQUIRENDAE

Amplistomum papillatum, Cobbold, 1882.

Found in intestine of Elephas indicus, India.
Amphstomum tuberculatum, Cobbold, 1875.

Found in intestine of Bos taurus, India.
Amphistomum emarginatum, Diesing, 1839.

Found in intestine of Niciipithecus trivirgatus, Brazil.

CONCLUSION

As a result of an exhaustive examination of a very large collection of
material comprising in many instances some hundreds of specimens,
and of a careful and critical study of the monographs of Fischoeder and of
Stiles and Goldberger, the conclusion is reached that many of the species
described by the former, and all except one of those described by the
latter authors, are merely synonyms of earlier species. It appears to the
writer that the authors have fallen in error owing to the fact that they con-
fined themselves to the examination of limited material, in some cases to the
examination of a single non-gravid worm, or even in one or two instances
to that of a series of sections of a single specimen. It is only when a long
series of specimens is examined that one realises to what extent individual
Variations occur.

202

LIST OF AMPHISTOMES ARRANGED UNDER THEIR HOSTS

Host
Manatus exunguts
rf latirostris

Tapirus americanus

Equus caballus

a err
> mulus ; rt
Rhinoceros sp. (Rhodesia)
Phacochoerus sp. 4
(North-East Rhodesia)
Sus sp. (Annam.)
Dicotyles albtrostris
# labiatus
99 torquatus

Hippopotamus amphibius

Bos taurus

Bos taurus indicus

Bos urus

Bos. sp. (Pagan dwarf bull), Torin

Bos bubalus (Bison europaeus)

re $}
... Brumptia gigas
. Gastrodiscus aegyptiacus ...

PARASITE

. Chiorchis fabaceus

g . es eins asper

Cladorchis pyriformis

. Gastrodiscus aegyptiacus ...

Pseudodiscus collins
Gastrodiscus secundus St
" aegyptiacus ...

.. Gastrodiscoides homints
. Cladorchis giganteus

29 eth]

bP) IF
Taxorchis schistocotyle

. Paramphistomum gigantocotyle

. wagandt
buxtfrons
Cotylophoron cotylophorum ?
3 minutum

Carmyerius cruciformis
Paramphistomum pisum

. Paramphistomum cervi

explanatum
orthocoelium
C otylophoron cotylophorum
Stephanopharynx compactus
Gastrothylax crumenifer ...
Fischoederius cobboldt

be elongatus
Carmyerius gregarius
i spatiosus

Homalogaster poloniae
Amphistomum tuberculatum ?

. Paramphistomum cervt

orthocoelium
Cotylophoron cotylophorum
Gastrothylax crumenifer ...
Fischoederius cobboldi

a elongatus

Carmyertus spatiosus
Paramphistomum explanatum

cervt
Cotylophoron cotylophorum
Paramphistomum cervt
Carmyerius gregarius

LocaTION

. Intestine.

> at
. Stomach.

. Intestine.
. Stomach.

. Large intestine
... Intestine.
... stomach.

99
>
29
2?

”

ian si Pas tite
. Stomach.

Bos (bubalus) caffer, Africa
Bos (bubalus) bubalis, Asia

Palonia frontalis ...

Anoa depresstcornts
Capra hircus
»» sp. (India)
» sp. (Northern Territory,
Gold Coast)
Ovis aries

fe ope. OFt Said)

,, sp. (South Africa) ...
» sp. (Hong Kong)

Antilope dorcas ...
Antilope sp. (Kamerun) ..

Cobus sp. (North-east Rhodesia)

Cobus sp. (Z eref)
Cobus maria "
Tragelaphus scriptus

- speket
Hi ippotragus equinus
Aepyceros melampus Se
Bubalus sp. (Nyasaland)

Bubalis sp. (Rhodesia)
Cervicapra sp. (Rhodesia)
Portax tragocamelus
Cervus alces

95 campestris.

a capreolus ...

», dama

» dichotomus

», elaphus

Ee MeX1Canus...

» namby

» rufus
simplicicornis

Elephas indicus

Loxodon africanus

Castor fiber

... Pseudodiscus hawkestt
. Brumptia gigas

203

. Cotylophoron cotylophorum

Carmyertus gregarius

. Paramphistomum cervi

Carmyerius gregarius

. Fischoedertus cobboldi

a elongatus
Homalogaster paloniae

. Fischoederius elongatus
. Paramphistomum cervi
.. Gastrothylax sp. ? (immature)

Paramphistomum sp. ?
(immature)

... Paramphistomum cervi

Cotylophoron cotylophorum .
Gastrothylax sp. ? (immature)

. Paramphistomum sp. ?

(immature)

: Paramphistomum orthocoelium

Gastrothylax crumenifer ..

... Parampbhistomum cervi

. Carmyerius spatiosus
Stephanopharynx compactus
Cotylophoron cotylophorum

. Cotylophoron cotylophorum
. Carmyerius wenyont
. Carmyertus spatiosus

. exoporus

. Paramphistomum cervi
. Carmyertius spatiosus

. Cotylophoron cotylophorum

>) bP)
Paramphistomum explanatum

.. Carmyertus spatiosus

bP) 9

. Paramphistomum cervi

rs cervt

7" liorchis

oS Cervt

>] 29
liorchis

Balanorchis anastrophus ...
Amphistomum lunatum ?...

. Paramphistomum cervt

liorchis

> 9

Amphistomum apillatom | ?

. Cladorchis subtriquetrus ...

. Stomach.

. Caecum.
. Stomach.

... stomach?
. Stomach.

PP]

... Intestine.
. Stomach.

vb}

. Intestine.

: Small and large

Intestine.

204

Callithrix noctivaga oe ... Amphistomum emarginatum? ... Intestine.
Cercopithecus callitrichus ... Pseudodiscus watsont S ... Colon.
Macacus cynomolgus... ... Pseudodiscus watsoni? ... ... Colon.
Homo sapiens... ve ... Gastrodiscoides hominis ... po Litestige.

Pseudodiscus watsont

REFERENCES

Braun, M. (1g11). Review of Stiles and Goldberger (1910) Zool. Zent., Vol. XVIII, p. 705. Leipzig.

Coxszorp, T. S. (1875a). On the Destruction of Elephants by Parasites ; with remarks on Two New
Species of Entozoa and on the so-called Earth-Eating Habits of Elephants and Horses in India.
The Veterinarian, Vol. XLVIII, October, p. 733. London.

——— (1875b). Further Remarks on Parasites from the Horse and Elephant, with a Notice of New
Amphistomes from the Ox. Jbid. November, p. 817.

Coun, L. (1904). Helminthologische Mitteilungen. II. <Archiv. fur Naturg. Seventh Year.
Vol. I, p. 229. Berlin.

Dapay, E. von (1907). In siidamerikanischen Fischen Lebende Trematoden Arten. Zool. fabrb.
Abt. Syst. Vol. XXIV, p. 467. Jena.

FiscHoEpER, F. (1g01). Die Paramphistomiden der Saugethiere. Zool. Anzeig., Vol. XXIV, p. 367.
Leipzig.

—~—— (1902). Die Paramphistomiden der Saugethiere. Jnaug. Diss. K6nigsberg.

——— (1903). Die Paramphistomiden der Saugethiere. Zool. Fabrb. Syst., Vol. XVII, p. 485. Jena.

——— (1904). Beschreibung dreier Paramphistomiden Arten aus Saugethieren. Jbid., Vol.
XX. Jena.

Innes, J. A. (1912). Gastrothylax bubalis, n.sp., with a few notes on the genus Gastrothylax (Poirier).
Parasitol., Vol. V, p. 271. Cambridge.

*Kosayasul, H. (1915). Watsonius macaci, n.sp., from caecum of Macacus cynomolgus. Dobuts. Z.

Tokio.
(1920). Digenetic Trematodes in Japan. Parasitol., Vol. XII, p. 392. Cambridge.

Leirer, R. T. (1908). An account of some Helminths collected in the Sudan. Welcome. Res. Lab.,
Third Report, p. 187. Khartoum.

(1910). The Entozoa of the Hippopotamus. Proc. Zool. Soc. Lond., p. 233. London.

(1913). Observations on certain Helminths of Man. Trans. Soc. Trop. Med., Vol. VI,
p- 265. London.

Looss, A. (1896). Recherches sur la Faune Parasitaire de l’Egypte. Mém. de I’Inst. Egypte, Vol. III,
Pt, 1. ‘Catre.

(1912). Ueber den Bau einiger anscheinend seltener Trematoden Arten. Zool. Fabrb.
Supp. XV., p. 323. Leipzig.

MacCativm, G, A. (1917). A New Trematode Parasitic in Elephants. Bull. Amer. Mus. Nat,
Hist., Vol. XXXVII., p. 866. New York.

Opuner, T. (1911). Zum natiirlichen System der digenen Trematoden: I. Zool. Anzeig., Vol.
XXXVII, p. 181. Leipzig.

Pirters, A. W. N. (1922). Paramphistomum cervi (Zeder) from a Cow in Cheshire. Vet. Fourn.,
Vol. LXXVIII, No. 8, p. 292. London.

Raituiet, A., Henry, A., and Joyeux, C. (1912). Sur deux Trématodes de Primates. Bull. Soc.
Path. Exot., Vol. V, p. 833.

Rarcuter, A., Henry, A., and Baucug, J. (19144). Sur les Amphistomiens des Ruminants domestiques
de PAnnam. Rec. Méd. Vét., Vol. XCI, p. 195. Paris.

* This paper was not consulted.

205

Rariuret, A., Henry, A., and Baucue, J. (1914b).
Bull. Soc. Path. Exot., Vol. VU, p. 79. Paris.

StePHENS, J. W. W. (1906). Note on the Anatomy of Gastrodiscus hominis.
Rep. (new ser.), Vol. VII, Pt. I, p. 9. Liverpool.

Stites, C. W., and GorpBerceR, J. (1908). Observations on Two New Parasitic Trematode Worms :
Homalogaster philippinensis n.sp., Agamodistomum nanus n.sp. U.S.A. Treas. Dept., Hygienic
Lab. Bull., No. 40, Pt. 3, p. 21. Washington.

Sur les Helminthes de I’Eléphant d’Asie.

Thompson Yates Lab.

(1910). Study of the Anatomy of Watsonius (n.g.) watsoni of Man and of Nineteen

Allied Species of Mammalian Trematode Worms of the Super-family Parampbistomoidea.
Ibid., No. 60. Washington.

SrunKarD, H. W. (1917). Studies on North American Polystomidae, Aspidogastridae, and Param-
phistomidae. Ill. Biol. Monographs, Vol. III, No. 3, p. 60. Urbana.

Travassos, L. (1921). | Contribugio para a systematica dos ‘ Paramphistodae,’ etc. Braz.-Med.
Anno XXV., p. 357. Rio de Janeiro.

206

EXPLANATION OF PLATE V

Paramphistomum cervi. Photographs showing variations in size and

shape exhibited by 24 specimens. x 24.

nals Trop. Med. &} Parasitol., Vol. XVII PLATE V

Fic. A

4 Brown C. Tinling & Co., Lid., Imp.

THE LIBRARY
| OF THE
HMIVERSITY ng ILLAMOIS

ABIVFANITY OF fit jeans

208

EXPLANATION OF PLATE VI

Fig. A. Paramphistomum explanatum. xX 2}.

Fig. B. Cotylophoron cotylophorum. x 2%.

Annals Trop. Med. & Parasitol., Vol. XV 11 PLATE V1

C. Tinling CF Con Lid. Imp.

Tht A pu hs

gf THE
QBIVERSITY oF Hb amutd
: a
7

210 &

EXPLANATION OF PLATE VII

Fig. A. Carmyerius exoporus, n.sp. X 2}.

Fig. B. Stephanopharynx compactus. X 2}.

Annals Trop. Med. & Parasttol., Vol. XVII PLATE VI1l

Fic, A

iter ge

M. Brown Ge Leniing ce *.CO gtd. 1 wip

THE WIeGAaY
OF THE
UNIVERSITY OF Hus

212

EXPLANADION, OF PLATHRSVitl
Brumptia gigas
Figs. 1 and 2. Dorsal view.

Figs. 3 and 4. Lateral view.

Figs. 5 and 6. Ventral view. (5) Genital papilla extruded ; (6) genital
papilla retracted. x 24.

4nnals Trop. Med. & Parasitol., Vol. XVII ATO ITT

| val .
M. Brown C. Tinling & Co., Ltd.. Imp

THE LIBRARY
OF THE
TRINERSITY OF WLAMOS

MALARIA IN AUSTRALIA

BY
P. A. MAPLESTONE

(Received for publication 25 February, 1923)

The following is a short account of the history of malaria in Australia
as far as it can be collected from the published records. In compiling
this review all the information prior to the year 1912 has been taken
from Cleland (1914); since 1912 the original articles in the medical
press and the various Government reports have been consulted. This is
not a complete discussion of all the references to malaria in Australia ;
many of the earlier accounts are given by laymen, or are merely the
-expression of an opinion by a medical man without definite proof, and
a considerable number of the later records are purely of local interest,
so they have been ignored. In discussing the subject, the various States
and the Northern Territory of the Commonwealth are considered separately.

QUEENSLAND

According to Cleland (1914), the first account of malaria in Australia
by a medical man is that of White (1867). In all probability this is
the same outbreak as the one to which Elkington (1912) refers, and which
he concludes was introduced to Burketown by a ship from Java. - Cleland
mentions that for some years prior to 1885 there was extensive and
severe malaria in North Queensland. In support of this statement he
quotes the following authorities, viz. :—A’Hearne (1890) for Townsville,
Graham-Browne (1890) for Charters Towers, Hunt (1890) for Hughenden,
and James (1891) for Croydon. But from the quotations taken by

214

Cleland from these authors’ writings it is by no means clear that all the
epidemics included in this series were due, altogether or even partly,
to malaria. For instance, there is no way of finding out if all A’ Hearne’s
cases were malaria ; Graham-Browne’s description of the Charters Towers
epidemic is unlike malaria; Hunt obviously confuses typhoid fever
and malaria, and James under the term ‘ Gulf fever’ describes all his
febrile cases, some of which were probably malaria. For the same period
Jeffris-Turner says he only saw three cases of malaria in children in
Brisbane. Whatever these various outbreaks were, it is a striking fact
that since the advent of more accurate diagnostic methods, malaria has not
been recorded from any of the above towns, but enteric fever is fairly often
encountered there.

From Cleland’s account it appears that O’Brien (1908) was the first
to record finding the malaria parasites in Australia ; nearly all his cases
were simple tertian, but he writes of finding a few quartan and malignant
tertian. O’Brien’s observations were apparently made at Yarabah
mission station near Cairns. Breinl (1911), without quoting an authority
and after only a few months in the country, reported malaria to be
epidemic in ‘parts of Queensland,’ mentioning specifically Innisfail,
Cooktown, and Saxby River. Nevertheless, he evidently saw some
rhalaria, as he adds that the locally acquired cases were simple tertian,
and that the few cases of malignant tertian that he saw were infected
in Papua (=British New Guinea).

Elkington (1912) refers to a localised epidemic of malaria, which
occurred at Kidston on the Einasleigh gold field in 1910. There were
120 cases and 24 deaths in a population of 400. The outbreak was
investigated by Dr. Baxter-Tyrie, who concluded that the disease had
been introduced from New Guinea. From the latter’s report Elkington
concludes that malignant malaria and blackwater fever are endemic ;
there is no further reference to this ‘endemic’ centre in the medical
literature nor in the Government reports, therefore it is clear that
Elkington’s conclusion was premature. Neither is there any evidence
that malaria spread from Kidston to any of the surrounding camps.

Although a few cases of malaria undoubtedly occur annually in the
coastal districts of North Queensland, there is no way of finding out
their numbers. In the Annual Health Report for the State, acute malaria
first appears as a notifiable disease in the period Ist July, 1915, to 30th
June, 1916. The figures up to the present time are given in Table I.

215

Taste I,

Cases of malaria notified in Queensland.

Year | No. of cases

1915-1916 ah Jee so wis E vee) a eee +f ne 79
|

1916-1917 | 213
|

1917-1918 an se 1 7 He ew + cae fe oe 72
|

1918-1919 “eh 45 dea te Be O| date az bok te Te)

IgI1g—1g20 + aM ne: a3. oy ove #22 LAs ee ae 9

1920-1921 9

1921-1922 = oe — nee ae «ant Pe sae at ae 19

Unfortunately there is no way of ascertaining how many of the above
cases were contracted in Queensland, and how many came from elsewhere,
except that in 1916-1917 the Sanitary Inspector of the Northern District
states in his report that 119 cases of malaria occurred in Cairns for that
year. It can also be found indirectly, by comparing the above figures
with those of the Australian Institute of Tropical Medicine for corresponding
periods, that nearly all the remaining cases were returned soldiers who
had become infected outside Australia.

Although this information is very incomplete it is quite obvious
that the Cairns epidemic was short-lived and that not many cases can
be occurring there at the present time. A possible explanation of this
short epidemic in Cairns is that this is the first port of call for boats
coming from New Guinea to Australia. In the period immediately
preceding and during the sudden increase in malaria in this town, large
numbers of soldiers returning from New Guinea were calling there, and
the majority of them were being sent home because they were suffering
from malaria. The extra opportunity for the mosquitoes of Cairns
to become infected soon reacted on the local inhabitants ; but in 1918,
when traffic between New Guinea and Australia returned to normal and
fewer persons with malaria parasites in their blood were calling at Cairns,
the incidence of malaria there suddenly dropped and has remained low
ever since. It is true that Breinl and Taylor (1918), after a malaria
and mosquito survey of the town recommended the filling and draining

216

of various swamps in and around it ; but the drop in malaria incidence
cannot be explained in this manner, because Mr. Hill, Entomologist of the
Australian Institute, visited Cairns in 1g2t and at the writer’s request
examined the mosquito-breeding places recorded by Breinl and Taylor
in 1918. He reported that little had been done in reducing these breeding
places.

Dr. H. H. Willis, in a letter to the writer in May, 1921, informed
him that while on a ‘ hookworm’ survey of the native settlement on the
Palm Islands he had found nine or ten* cases of acute malaria which
he had diagnosed microscopically. Over a month later the writer visited
these Islands ; he examined all the natives (over 300) and found that
five or six* of the cases reported by Willis had crescents in their blood.
All the other natives were negative on blood examination, no palpable
spleens were found although there were numerous children, and no fresh
cases had occurred between the visits of Willis and the writer. The
evidence of the origin of this small outbreak was not satisfactory, but as
far as could be gathered it seemed likely that the malaria had been intro-
duced from the mainland by some recent arrivals. It is remarkable
that the outbreak did not spread further, because the natives were living
closely congregated in unscreened grass huts, and Hill found Anopheline
mosquitoes breeding close to the dwellings (see Table VI).

The history of Townsville during recent years from the point of view
of malaria is of considerable interest, because since the establishment of
the Australian Institute of Tropical Medicine in 1910, more reliable records
are available from there than from any other town in Northern Australia.
Many parasite carriers have been constantly arriving in Townsville
for treatment during the eleven and a half years January, Ig1o to
June, 1921, and for the whole of that time no case of malaria has ever been
discovered which was contracted in the town or its surroundings. There is
also abundant evidence that the same species of Anophelines are found as
in other Coastal towns where malaria occurs. Townsville is well within
the tropics and is by far the largest town in Northern Australia with
a population of about 25,000, nearly all of whom are whites.

* Figures from memory.

217
THE NORTHERN TERRITORY

Again consulting Cleland (1914) it is found that Wood (1889) said
that malaria was very prevalent in the Northern Territory during the years
1879, 1880, and 1881. Holmes (1913) gives the following figures taken
from the official records :—

Taste IT.

Deaths in the Northern Territory.

Year | Total Number of deaths Deaths due to ‘ fever’
1579. ... oe eis at ais ae 166 61
Decne E. te wes i, Fe i 154 61
BOSE fo (0: vT 43: =. tele of 100 SI

It was at this time that gold-mining was at its height and there were
many mining camps in the country with no medical man near them and no
sanitary precautions in force. Although many of the deaths were in all
probability due to malaria, it should be borne in mind that a large number
of the cases could not have been seen by a medical man, so that the
diagnosis of ‘fever’ on the death certificates is not of much use for
accurate record.

The Umbrawarra tin mining field was opened up about the year 1909
and shortly afterwards malaria broke out there. This epidemic was
authenticated by Breinl (1912) who gives an account of it. The record
is of considerable value, because the epidemic is shown to be due beyond
all doubt to malaria, and the conditions at Umbrawarra were, in all
probability, identical with those obtaining on mining fields in earlier
times where similar epidemics occurred. These rushes to new mineral
discoveries attract men from other parts of the world, and all the mining
camps of Northern Australia have contained men from New Guinea,
which is a highly malarious country. In the case of Umbrawarra, Breinl
traced the origin of the outbreak to miners from New Guinea arriving with
parasites in their blood; from this fact it seems most likely that the
earlier epidemics on other mining fields in the same regions of Australia
were due to a like cause. The Umbrawarra epidemic came to an abrupt
end, primarily by the departure of the majority of the miners, for in 1913,
when the writer visited the field, there were only two miners remaining ;

218

but it is strange that the disease did not spread to Pine Creek, a permanent
settlement only thirteen miles distant and which during the height of the
activities at Umbrawarra was in daily communication with it, receiving all
the men who were seriously ill and many of whom must have had malaria
parasites in their blood. The same species of Anopheline has been
recorded from both places.

Breinl and Holmes (1915) visited several districts in the Northern
Territory, including the Daly and Alligator Rivers, and Bathurst and
Melville Islands; they found no signs of malaria among the natives in
any of these localities either on blood examination or spleen palpation.
In the same report it is mentioned that Holmes in 1912 found four out of
twenty natives examined on Melville Island to be suffering from malignant
tertian malaria, from which it is clear that within three years the disease
had disappeared from the island without any special anti-malarial measures
being taken.

The only other published records of malaria in the Northern Territory
are those in the Annual Health Reports and in the Annual Reports of
the Darwin Hospital. A brief outline of the local conditions will indicate
that the figures in Table IV are only very approximate.

The total area of the Northern Territory of Australia is well over
500,000 square miles; there is only one* medical officer in the whole
country and he spends practically the whole of his time in Darwin. For
this reason most of the cases of sickness reported, and of deaths registered,
are not certified by a qualified man, but are furnished by the local police ;
consequently the only reliable returns are those for the Darwin Hospital.
In the year 1918 the highest population since 1910 was recorded, and
this is given in Table III along with the latest figures available.

TaBLe III.

Population of the Northern Territory.

Year Europeans Asiatics Half-castes Total
1918 i ta od og 8 Bis 3767 1177 118 5062
1g21 het nad =p Pe ps 2478 1094 ? 3572

* From 191i to 1915 there were four medical officers in the Northern Territory, and two of them
did considerable travelling.

219

The aborigines consist of numerous small nomadic tribes, hence their
numbers cannot be accurately determined. The most reliable estimate
of the number of natives that the writer has ever been able to obtain, was
given to him about ten years ago by an official who had spent over forty
years in the country and who had travelled practically all over it. This
officer was of the opinion that there were not more than 30,000 natives
in the whole country. Although far from being precise these figures at any
rate indicate that it is very thinly populated.

Tasce LV.

Malaria records for the Northern Territory

Year Total Number of Cases treated in Deaths Remarks
cases reported Darwin Hospital

1897...
1898...

1899...
1900...
1901...
1902...
1903...
1904...
1905...
1906...
1907...
1908...
1909...
1910...
RO1T...
cOl2..
OF
1914... aa oe ee
1915... mee aug As —

1916... 7 Bad aa ‘ Prevalent ’

-_
Qe NN we wr DO CO

my tt tte et
| — |
Om OW CMAN OO DADS An ON |

Pa =e NPN
~v An HFN FW N

For 18 months
ending 30.6.1917

_
as

1917... af A ae =
1918... A a ets 45
191g... ae ots 4% —
1920... ey es oa 59
1921... =F ame ...| © Many cases’

Not available

Rwvl~|
vwl ~|

In addition to the above table the following extracts from the Health
Reports are appended.

1912. Malaria is not as prevalent as it is popularly supposed to be.

The only death ascribed to malaria is registered ‘ kidney troubles and

fever.’ Practically all deaths outside Darwin are registered by the

police, so the accuracy of the diagnosis is extremely doubtful. Malaria

220

is unknown at Pine Creek and Darwin, the two largest settlements.
Some cases of malaria were found among the natives on Melville Island.

1913. The single death registered as due to malaria was diagnosed
by a layman. Two of the medical officers travelled extensively during
this year and only one case of malaria was seen, although this disease
was specially looked for, and no cases were found on Melville Island
where it was seen the year before.

1915-1917. Malaria was ‘very prevalent’ in several localities,
e.g., the Pine Creek railway extension camps and Maranboy mining
field. More than 50 per cent. of the cases were only diagnosed clinically,
and although it is not stated, it is almost certain that a number of the
cases were not seen at all by a medical man. It is considered that
malaria is not endemic.

1918. One case was contracted in Darwin.

Notre.—This is the only record of a case contracted in
Darwin that the writer can find.

1920. All of the 59 cases reported for the year came from the
country districts and were of a mild form. Three more serious cases
were apparently contracted elsewhere.

1921. Many cases have occurred during the past few months, none
of which were contracted in Darwin. The increase of the past few
years is ascribed to the introduction of returned soldiers with parasites
in their blood.

NEW SOUTH WALES

Early in the year Jamieson (1915) reported a case of malaria which
the evidence showed to have been contracted at Gosford not far from
Sydney. Commenting on this case, Cleland (1915) stated that apart
from unreliable records in the comparatively early days of settlement he
only knew of one other case contracted in the State. This was in a baby
a few days after birth, who was born of a mother suffering from malaria
at the time ; he considers this to be a case of direct infection.

On the 17th March, 1915, ‘ Acute malaria’ was made compulsorily
notifiable throughout New South Wales; this regulation continued in
force until 28th November, Ig19, when it was withdrawn. The annual
figures for this period are given in Table V.

221

TaBie V.

Cases of malaria notified in New South Wales.

Year Number of cases
1915 105
1916 61
1917 t7
1918 II
1919 35

It is not stated in these returns whether any of the cases were locally
acquired, and all that can be gathered in this respect is that in 1915 the
Chief Health Officer in his letter of presentation of the annual report
states, that of the 105 cases recorded in that year, all except 14 were
returned soldiers ; it is, of course, possible that all of the fourteen cases
who were not soldiers also acquired their infections in other countries.

There are two other records of isolated cases which seem beyond
doubt to have been contracted in New South Wales; one of these was
reported by Evans (1919) at Wyong, and the other by Clayton and Utz
(1921) near Tumbarumba. This completes the published record of
malaria for New South Wales.

VICTORIA
Doyle (1921) reported a case of malaria at St. Arnaud, which was
locally acquired. As far as can be ascertained this is the only case of
malaria ever recorded in Victoria.

In South Australia and Tasmania there is no evidence that malaria
has ever occurred. The North-west of Western Australia which adjoins
the Northern Territory 1s comparable to the latter both in its malaria
incidence and conditions of living. No references to malaria in this part
of the country can be found in the literature, nor are the Government
reports from this State available, so actual figures cannot be given.

The writer had spent altogether upwards of five years in North
Queensland and the Northern Territory (see map, places underlined),
and during that time he has seen only two cases of malaria contracted
in the country, the small outbreak on Palm Island in 1921 excepted.
Experience has led him to the conclusion, that the inhabitants of Tropical

222

Australia are prone to ascribe all their ills to malaria and that this opinion
is rarely confirmed by microscopic diagnosis.

The tendency of the layman to exaggerate the incidence of malaria
reacts on the police, who in the absence of medical assistance are inclined
to register all deaths not clearly due to violence as due to malaria. These
figures are given in the annual health reports and so the popular and
erroneous opinion of the prevalence of malaria is to some extent supported
in official returns.

ANOPHELINE MOSQUITOES FOUND IN AUSTRALIA

According to Ferguson (1921) only five species of Anophelines have
ever been recorded in Australia. They are :—

1. <A. corethroides, Theobald, 1907.
A. (Pyretophorus) atratipes, Skuse, 1888.
A. (Pyretophorus) stigmaticus, Skuse, 1888.
A. (Nyssorhynchus) annulipes, Walker, 1850.*
A. (Myzorhynchus) barbirosiris, de Wulp, var. bancrofit,
Giles, 1902.

copteiie “Spdb opt

A. corethroides is only found in South Queensland.

A. atratipes was recorded by Bancroft (1908) from South Queensland
and it was also recorded earlier by Skuse at Berowra, New South Wales.

A. stigmaticus has been recorded only once from a single locality
in New South Wales.

These three species are only found in parts of Australia where malaria
does not occur and are so restricted in distribution that they cannot have
any bearing on the spread of this disease at the present time. The other
two species, viz., A. annulipes and A. bancroftt are much more widely
spread and as, from circumstantial evidence which is all that is available,
they seem to be connected with malaria, their occurrence will be considered
in more detail.

Distribution of A. annulipes. Hill (1922) summarises
the distribution of this species as follows :—‘ A. annulipes is undoubtedly
the most widely-distributed Anopheline found in Australia, having been
recorded from Tasmania northwards to Banks Island (Torres Strait), and
from South Australia, Central Australia, Northern Territory, South-west
Australia, and North-west Australia. It is most probable that it does not
occur in the elevated districts of South Australia and North Queensland
(Atherton Tabieland), and possibly not in some of the arid inland districts,

* Until recently, another species Anopheles amictus Edwards, 1921, has been confused with
A. annulipes. A. amictus occurs at Townsville, Palm Island and Port Darwin. [Editors.]

223

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224

although its presence in such localities as Wire Creek Bore and Lander _
Creek (Central Australia), indicates that only the absence of suitable
breeding places would inhibit its existence in the latter.’
Distribution of A. bancroftt. This species is not so
Hill states that it is found only in the
coastal districts of Queensland, some of the Torres Straits Islands, and
He |

widely spread as A. annultpes.

Northern (coastal) districts of the Northern Territory
. but one of the localities given by Hill, viz., Horseshoe Creek is
over 150 miles from the coast. As it is possible that this species has
more to do with malaria in Australia than has hitherto been supposed,
it is proposed to consider the records of its occurrence in more detail.

These are summarised in Table VI.

Taare VI.

Principal records of the occurrence of 4. bancroftt in Australia.

Locality By whom recorded Remarks

Brisbane ... .. Bancroft (1908) Found in the scrub from Enoggera to
Caboolture; males, larvae, and eggs
never found.

Brisbane ... ... Cooling (1913) Once taken in a house; a few females
| taken in the scrub; no larvae found.

Brisbane ... , Cooling (1914) Four specimens taken for the whole year. |
|

Rockhampton . Taylor (1916) .| Numerous adults, no larvae.

Burdekin River...

Townsville

Halifax

Palm Islands

Cairns

Northern Territory

Taylor (1913)

... Taylor (1916)
se Hill (92%) eee

..-| Taylor (1916)

Taylor and Breinl (1918) ...

-| Hill (1922) ...

Hill (1922), writes that he has failed to find
the species over a period of 24 years’
continuous observation and _ concludes
that it has died out.

...| Plentiful within 200 yards of No 2 aboriginal

camp and close behind main camp.
A. annulipes not taken nearer than } mile
of camp clearing.

Numerous, and breeding freely in swamps
in and around town.

...| Twelve distinct localities given, including

Melville and Bathurst Island (see map).

Note.—The mosquito observations in the
N.T. are more thorough than in any part
of Tropical Australia except for a small
area round Townsville.

225

With the exception of Brisbane, Townsville, Cairns and to some extent
the Northern Territory, the records in Table VI refer to a single observation.
It is probable that more extended work would reveal the presence of
A. bancrofti in many other parts of Northern Australia, but it is unlikely
that this species exists far south of Brisbane, because it has never been
recorded in New South Wales at all, and the knowledge of mosquitoes in
this State is much more advanced than it is in Queensland. In fact,
as far as the records go, there is some evidence that A. bancrofti is not
found in any numbers south of about 19° South Latitude.

THE INSECT VECTOR OF MALARIA IN AUSTRALIA

The Medical Journal of Australia has on more than one occasion
(Leading articles, I915, p. I7I1 and 1921, p. 512, etc.), pointed out that
the mosquito carrier of malaria has not yet been determined. Breinl
(1912) stated that A. annulipes was the probable vector at Umbrawarra ;
in support of this he quotes the successful experiment of Kinoshita (1906),
who successfully infected A. annulipes with Plasmodium falciparum in
Formosa. A little later Breinl (1914) definitely stated that A. annulipes
was the carrier of malaria in Australia, but quoted no authority. Since
these two references A. annulipes has been frequently mentioned in the
medical literature of Australia, being variously described as ‘ the probable
carrier,’ ‘the presumed carrier,’ ‘ the carrier, etc., but in most cases no
authority is given, and when it is, Kinoshita (1906) is the only reference.
The statement by Harrison (1922) is an accurate summary of the present
state of our knowledge in this respect, when he says that there is evidence
that the local Anophelines are capable of acting as intermediate hosts for
malaria parasites.

The only record of A. annulipes as a malaria carrier given by Chanal
(1921), is the single experimental result obtained by Kinoshita already
referred to; Chanal’s conclusion is that A. annulipes should be classed
as dangerous. But it should be noted that although Kinoshita states
that he infected 60 per cent. of his mosquitoes, a detailed study of his
experiments shows that he used the species on three occasions. The
first time, out of five fed, all died within three days, the second time nine
insects were used and all died within three days of feeding, the third
time eight mosquitoes were used, of which three died within three days

226

and three of the remaining five became infected. It is this result Kinoshita
gives as 60 per cent. positive ; only P. falciparum was used.

A. bancroftt seems by common consent to have been almost completely
ignored as a possible malaria catrier in Australia, for the only references
to this species in this connection are the following, viz. :—Cleland (1g10)
includes it in a list of the then known malaria carriers. It is next men-
tioned by Cooling (1914) who suggests that it may be a malaria carrier,
because Stephens and Christophers (1902) were successful in infecting
A. barbirostris in the laboratory in India. Since that date, according to
Chanal (1921), A. barbirostris has been found in nature and infected in the
laboratory both with P. falciparum and P. vivax on several occasions in
various parts of the Malay Archipelago ; but as all these records refer to
a different variety of the species which does not occur in Australia, they
have no bearing on the subject. One other reference to A. bancrofti as
a malaria carrier is made by Breinl (1915) who, in an article on New
Guinea, states that Nyssorhynchus bancrofti* is not a malaria carrier.
This statement is not supported by any evidence.

With regard to the distribution of Anophelines in Australia, Breinl
(1914) says,“. . . The distribution of malaria in Australia corresponds,
on the whole, with the incidence of the mosquito Nyssorhynchus annulipes

It is curious to note that there are localities where the mosquito
has been found, but where malaria is practically non-existent.’

Again, Breinl and Taylor (1918) remark, *. . . Nyssorhynchus
annulipes which, judged by its distribution in relation to malarial infested
regions in Northern Australia, most probably acts as a malaria carrier

These two statements may be more or less correct as far as they go,
but they do not explain why malaria is practically never found far south
of Cairns, whereas A. annulipes is spread all over Australia. If the
explanation of the restriction of malaria to Northern Australia is to be
found in the distribution of a special Anopheline, it will be found that
thesoccurrence of A. bancroftt much more nearly corresponds with the
malaria distribution than does A. annulipes. A. bancrofti, however, also
exhibits one or two striking exceptions to the rule, so it is considered that
the peculiar distribution of malaria in Australia is due to other causes,
not yet ascertained.

* Myzorhynchus bancrofti is obviously the species intended, for Taylor (1914) in the list of mos-

quitoes taken by Breinl on this expedition includes it, and as far as the writer can ascertain there
is no such species as Nyssorbynchus bancrofti.

227

As far as the writer can find out, the only explanation that has ever
been offered as to why malaria fails to become established in the greater
part of Australia is the mathematical hypothesis of Ross (1910) ; all the
authors who mention this subject are of the opinion that there are too
few mosquitoes or too few susceptible human beings in most parts of
Australia. It is unlikely this is the sole reason, if it is the reason even in
part, for it is by no means in the most thickly populated parts of Australia
where Anopheline mosquitoes are found that malaria outbreaks occur.

There is another set of conditions which seem to the writer worthy of
consideration, and which have never been considered, and that is the
relation between malaria outbreaks and meteorological records. Gill
(1920 and 1g21a) has studied the incidence of malaria in parts of India
along with the mean temperature and relative humidity readings, and
the same author (1g2Ib) extended his observations to England. As a
result of this work he considers it probable, that before malaria is able to
spread in a locality it is necessary to have a monthly minimum mean
temperature of 61° F. anda minimum mean relative humidity of 63 per cent.
At the same time he points out this is not yet conclusively proved. In
this connection it is worth noting that in Kinoshita’s successful experiment
with A. annulipes the temperature remained between 28° and 30° C. the
whole time, and in his conclusion he states that complete development of
the oocysts of P. falciparum cannot take place in this mosquito except in
a high and unvarying temperature.

SUMMARY

As far as can be gathered from the incomplete and unreliable records
available, malaria is only mildly endemic in Australia north of 19° South
Latitude. <A. annulipes and A. bancrofti the only two possible malaria
carriers in Australia under present conditions are much more widely
distributed than is malaria.

In various localities north of 19° South Latitude small epidemics of
malaria occur from time’to time ; these outbreaks are of short duration,
their origin is generally traceable to the introduction of malaria carriers
from abroad, the disease does not spread to adjoining camps and towns,
and soon dies out, without any very active anti-malarial measures being
instituted.

228

The scarcity of population and Anopheline mosquitoes is not a satis-
factory explanation of the absence of malaria from the greater part of
Australia.

It is of the first importance to discover the mosquito carriers of malaria
in Australia, and when this has been done, work along the lines of Gill
in India and England would possibly yield interesting and valuable results.

REFERENCES

*A’Hearng, J. A. (1890). Presidential Address to the North Queensland Medical Association. Aust.
Med. Gaz., August, p. 293.

BancrorT, T. L. (1908). List of the Mosquitoes of Queensland. Ann. Old. Mus., No. 8, p. 10.
Brent, A. (1911). Report of Aust. Inst. Trop. Med. for 1910, p. 21.

(1912). Report on Health and Disease. Bull. No. 1a, Northern Territory, p. 11.

——— (1914). The Distribution and Spread of Disease in the East. Stewart Lect. No. 1, Univ.
Melb., 1913.

——— (1915). On the Occurrence and Prevalence of Diseases in British New Guinea. Ann. Trop.
Med. & Parasitol., Vol. TX, p. 289.

Brent, A., and Hoimes, M. J. (1915). Medical Report on the data collected during a journey
through some districts of the Northern Territory. Bull. No. 15, Northern Territory.

BreEIn1, A., and Taytor, F. H. (1918). A Malarial Survey of the Township of Cairns. Med. Fourn.
Aust., Vol. II, p. 109.

*Browne, G. (1890). Aust. Med. Gaz., p. 322.

Cuanat, L. (1921). Réle Pathogéne des Moustiques, etc. Travail du Lab. de Parasit. de la Fac.
de Méd., pp. 13 and 14.

Crayton, J. H., and Urz, L. (1921). A Case of Malaria infected in the Riverina, New South Wales.
Med. Fourn. Aust., Vol. I, p. 382.

CLELAND, J. B. (1910). Flies and Disease. Rep. Govt. Bureau, Microbiol. N.S. Wales for 1909, p. 52.

(1914). Contributions to the History of Disease of Man in Australia. Ibid. 3rd Rep.,
p- 226.

(1915). Malaria in New South Wales. Med. Fourn. Aust., Vol. I, p. 316.

CooxtneG, L. E. (1913). Report on Mosquito Work in Brisbane. Append. F., Ann. Rep. Commr.
Pub. Health, Queensland, p. 59.

(1914). Second Progress Report of the Campaign against Mosquitoes in Brisbane. Append.
H. Ibid. (1914), p. 63.
Doyte, G. (1921). Clinical Reports of Two Unusual Cases. Med. Fourn. Aust. Vol. 1, p. 421.
ELkiNnGTON, J. S. C. (1912). Quarantine in Australia. Aust. Med. Gaz., p. 434.

Evans, W. (1919). Anti-malaria Work with the Australian Mounted Division in Palestine. Med.
Journ. Aust., Vol. II, p. 529.

Fercuson, E. W. (1921). The Malaria Danger. Med. Fourn. Aust., Vol. I, p. 432.

Gi1t, C. A. (1920). The Relationship of Malaria and Rainfall. Ind. Fourn. Med. Res., Vol. VII,
p- 618.

——— (1921a). The Rédle of Meteorology in Malaria. Ind. Journ. Med. Res. Vol. VIII,
p- 633.

——— (1921b). Malaria in England with special reference to the Réle of Temperature and
Humidity. ‘Fourn. of Hyg., Vol. XIX, p. 320.

* Original papers not consulted.

229

Harrison, L. (1922). Flies and Infectious Disease. Med. Yourn. Aust., Vol. Il, p. 490.

Hitt, G. F. (1922). Notes on the Habits and Distribution of some North Australian Culicidae.
Commonwealth Health Dept. Service Pub. No. 21.

Homgs, M. J. (1913). Report of the M.O.H. for Year 1912. Rep. Adminis. N. Terr., p. 53.
*Hunt, J. S. (1890). The Evolution of Malaria. Aust. Med. Gaz., p. 75.

*James, P. (1891). Remarks on the Fevers and Diseases of Tropical Queensland. Jbid., p. 300.
Jamigson, S. (1915). Malaria arising in a non-malarial District. Med. Fourn. Aust., Vol. 1, p. 163.
*Jerrris-TurNER, A. (1890). Aust. Med. Gax., p. 65.

Krnosuita, K. (1906). Uber die Verbreitung der Anophelen auf Formosa und deren Beziehungen
zu den Malariakrankheiten. Archiv. f. Schiffs.-und Tropenbyg, Vol. X, p. 714.

*O’Brien, R. A. (1908). Notes from North Queensland. Just. Med. Gaz., p. 121.
Ross, R. (1910). The Prevention of Malaria.

STEPHENS, J. W. W., and Curistopuers, S. R. (1902). The Relation of Species of Anopheles to
Malaria Endemicity. Reports to the Malaria Comm. Roy. Soc.

Taytor, F. H. (1913). Report of the Entomologist. Aust. Inst. Trop. Report for 1911, p. 49.
(1914). Culicidae from Papua. Trans. Ent. Soc. Lond., Pt. 1.

(1915). Report for the Half-year ending June 30th. Aust. Inst. Trop. Med. Half-yearly
Rep., p. 20.

(1916). Report of the Survey of the Distribution of Stegomyia fasciata in the Ports of
Queensland. Aust. and Yellow Fever. Quarantine Ser. Pub., No. 6, p. 57

(1917) Malaria Mosquito Survey of the Irrigation areas in the Murray River District
Ibid., No. 12.

*Wuire, J. A. (1867). On the Fevers of the Gulf of Carpentaria. Aust. Med. Gaz., Vol. XII, p. 361.
*Woop, P. M. (1889). Intercolonial Medical Congress Report, p. 54.

* Original papers not consulted.

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231

COCCIDIOSIS OF CATS AND DOGS AND
THE STATUS OF THE, ISOSPORA OF
| MAN

BY
C. M. WENYON

WELLCOME BUREAU OF SCIENTIFIC RESEARCH

(Received for publication A pril 13, 1923)
PLaTEs I[X-XIV.

Our knowledge of the coccidia of dogs and cats commences
with certain observations recorded by Finck (1854) on the changes
undergone by the intestinal epithelium of cats during the process
of food absorption. From that time to the present day it has been
generally assumed that these animals harbour only one coccidium,
which has been usually described during recent years under the
name /sospora bigemina (Stiles, 1891). Though the measurements
of the oocysts given by various observers who have studied the
coccidia of these animals have differed considerably, the view that
only one form exists has been rigidly adhered to with few exceptions.
Perroncito (1882) appears to be the first to have considered it
possible that more than one form occurred in these animals, for he
separates the one described by Grassi (1879) from that recorded by
Rivolta (1874-1878), while Neumann (1888), Railliet (1895) and
Neveu-Lemaire (1g12) seem to have held the same view. Dobell
(1919, p. 177) states that there is no really conclusive evidence to
prove that the /sosfora of the cat 1s the same as that of the dog, or
that both are merely varieties of one species, but he refers to all the
coccidia of these animals by the name /sospora bigemina, pointing
out, however, that Grassi’s name Coccidium Rivolta has priority
over that of Stiles. Reichenow (1921) definitely asserts that the
form in the dog is probably distinct from that in the cat, while
Noller (1921), without giving any details, writes of a small and a
large form in the cat.

Observations which have been made by the writer during the

Z52

past twelve months reveal the fact that there are at least three
species of /sospora in these animals in England. One of these has
an oocyst about 12-15 microns in length, another an oocyst about
25-30 microns in length, and a third an oocyst about 40-45 microns
in length. The text-fig. 1 shows the relative size and appearance
of the three types compared with the one discovered in man during
the war. If these dimensions are kept in mind, the different
accounts which have been given by various observers become at once
intelligible, and it is possible to identify with some degree of
certainty which form was actually under observation. All three
have been previously recorded in the literature. In addition to the
species of /sospora, dogs harbour an Ezmeria with which we are not
for the moment concerned.

HISTORICAL REVIEW OF LITERATURE

The first description of a coccidium of the cat was published by
Finck (1854). His paper is difficult to obtain, but fortunately
Davaine (1860) quotes in full the passage dealing with the bodies
observed by this author. As it is of such importance from the
present point of view it is quoted zz extenso from Davaine,

pp. 259-260.

“Sur le méme animal (le chat) nous avons rencontre une autre forme bien plus
singuliére (fig. 22). Beaucoup de villosités, semblables du reste a celles chargées de
graisse, a la place de gouttes graisseuses, renfermaient, en quantité considérable, des —
corpuscules que nous appellerons géminés, parce que le plus souvent ils étaient
réunis par paires. ‘T'antdt une seule et méme villosité offrait a la fois et des
gouttes huileuses manifestes et des corpuscules géminés, le tout entremélé d’une
maniére irréguliére ; tantot les corpuscules géminés remplissaient seuls le bout de
la villosité. Ils étaient pour la plupart elliptiques, et leur grand diamétre atteignait
4 peine un centiéme de millimétre ; la plupart mesuraient O™™, O8 sur O™™, O7,
ou bien O™™, 1 sur O™™, OO. Leur contour était fin, net, trés fioir'y leur
contenu variable, occupant tantdt presque toute la cellule, plus souvent accumulé
vers son centre. C’était une matiére granuleuse réunie en une ou plusieurs masses.
Tl nous a semblé parfois voir une enveloppe commune pour deux corps géminés.

” Quel est la nature de ces corps ? Remak représente un corpuscule semblable
au premier aspect, seulement plus grand et non géminé. I croit devoir le con-
sidérer comme un parasite particulier qui se développerait dans les cylindres
épithéliaux des glandes de Lieberkiihn et dans ceux des conduits biliaires. I
cite Hake et Nasse comme ayant trouvé des formes semblables, par masses, dans
le foie du lapin. Kdlliker a observé la méme chose. Selon lui, les corpuscules
du foie du lapin seraient des oeufs de bothriocéphale ; ceux des villosités du méme
animal, plus petits que les premiers, des oeufs d’entozoaires, siégeant dans |’intérieur
des villosités et peut-étre aussi dans les cellules épithéliales distendues. Dans ce

Fic.

a8 be

1. Diagram of the oocysts of the Isospora of cats, dogs and men. XX 2000.
Oocyst of the small form which occurs in the deeper tissues of the villi of cats and
dogs and man (Isospora bigemina and Isospora hominis).

Oocyst of the intermediate sized form which occurs in the epithelium of the villi
of cats and dogs (Isospora rivolta).

Oocyst of the large form which occurs in the epithelium of the villi of cats and
dogs (Isospora felis).

Oocyst of the large form which probably occurs in the epithelium of the villi of
man (Isospora belli),

234

dernier cas, ils ressemblent, selon lui, 4 des grosses gouttes graisseuses remplissant
les cellules épithéliales. |

” Nous n’avons rien trouvé de pareil dans les cellules épithéliales de notre chat ;
mais son foie renfermait des amas d’entozoaires plats, elliptiques, long d’un
millimétre, probablement des douves. [Ils étaient contenus dans des espéces de
kystes.

> Quant a nous, tenant compte de l’énorme quantité des corpuscules en question,
de l’absence de toute forme semblable dans la cavité de l’intestin, de leur absence
dans toutes les villosités n’ayant point subi l’espéce de macération caractérisant
les villosités farcies de globules graisseux, enfin de certaines formes de transition
entre ces derniers et les globules géminés, nous croyons ne pas trop nous hasarder
en rattachant les corpuscules en question au fait du méchanisme de l’absorption
graisseuse. C’est tout ce que nous pouvons en dire quant a présent.”

(Henri Finck : Sur la phystologie de Pépithélium intestinal, 'Thése de Strasbourg,

ESSA, 2° saetie: tee 424," p. 17).

The important points to note from thé above description are
these. The sporocysts or corpuscules géminés, as Finck styled
them, occurred in the substance of the villi and not in the epithelium
of the cat’s intestine. They measured 8 by 7 microns up to 10 by
g microns, had definite contours, and were sometimes enclosed in
pairs in a common membrane which was evidently the oocyst wall.
As pointed out by Railliet and Lucet (1891), Finck’s measurements
have been wrongly quoted as ten times higher than they actually
were by several observers, as, for instance, Pfeiffer (1890, 1891),
Neumann (1892, p. 467). Dobell (1919) inadvertently refers to
Finck’s investigations as having been made on the dog, instead of
the cat.

The reference to the similar but larger bodies seen by Remak,
referred to by Finck, have to do with a paper by this author
published in 1845 on the occurrence of what were evidently the
oocysts of a coccidium in the intestinal wall of the rabbit. Vulpian
(1858) cites Finck’s observations, but it is not quite clear that he
actually observed the oocysts of the cat coccidium himself. Rivolta
(1873, p. 382), referring to the presence of psorosperms in domestic
animals, says that they had previously been observed by Finck
(1854), and also by Ercolani in 1859, in the cat. Perroncito (1882)
also quotes Ercolani as having made this observation. Virchow
(1860, p. 342 and p. 527) was the next observer to give any details
of their structure, though, like Finck, he regarded them as products
of fat absorption. He noted that the villi of the greater part of the
intestine of a dog were infiltrated with psorosperms. They were on
the surface of the intestine, but a larger number were free in the

235

intestinal contents. They occurred in the interior of the villi and
were relatively small and regularly arranged in pairs enclosed by a
double contoured membrane. He says they were evidently similar
to the paired bodies described by Finck from the cat. He records
and figures the oocysts of a coccidium which he found in the kidney
of a bat, and which he regarded as similar to the one seen by him
in the dog. The parasite of the bat evidently belongs to the genus
lsospora.

Leuckart (1860, p. 11, and 1866, p. 21) mentions the fact that
the intestinal mucosa of a dog which had been used for experiments
with Z7ichina was much altered, and covered with a layer of small,
egg-shaped psorosperms. He gives no details of their size or
structure. He again (1863) refers to them, but is inclined to regard
them as metamorphosis products in the intestinal wall. Another
reference to these bodies found in another dog by the same author
(1879, p. 282) gives no further information, but he was aware of
Finck’s work and evidently regarded the structures he had
encountered as the same as those seen by Finck. He now describes
the condition as due to an accumulation of parasites in the vill.

,Rivolta (1874) gave a description of certain oviform cells
(cellule oviformz) which he had found in the intestinal villi of dogs
and cats. In his account, which deals entirely with those seen in
dogs, he says they had walls showing a double contour, and varied
in length from 8 to 12 or even 15 microns; while in breadth they
measured 8 microns. The contents of some of the oviform cells are
described as being granular and in the form of a nucleus, or as an
elongated body like an embryo with granular material at its centre.
In some, however, it is stated that in addition to a granular nucleus
there were distinctly three or four elongated corpuscles somewhat
irregular in shape. There are four figures accompanying this
description, and two of these show quite clearly the granular mass
and four small ovoid bodies. The oviform cells are described as
occurring in the tissues of the villi especially near their tips and not
in the epithelium. As evidence of this, a case is quoted where they
were present in the villi of a dead animal which, owing to cadaveric
changes, had lost its intestinal epithelium entirely. These oviform
cells are again described by Rivolta (1877). In this paper mention
is made of Finck’s observations, and it 1s pointed out that invasion

236

by the cells produces grave alterations in the structure of the villi.
In a further communication, Rivolta (1877a) states that he has found
other cases of the infection in dogs. Examination of the ovitform
cells in Miiller’s solution showed that they constantly contained
four long corpuscles with rounded ends. Two other stages are
described and figured showing the bodies filled with a granular
mass which may have indications of a central constriction. He
ventures the suggestion that proliferation into two is_ taking
place. The figures show this clearly. The length of the bodies
is given as 13 to 16 microns, and the breadth as 12 microns. —
The statement is made that they are identical with the corpuscules
géminés observed by Finck. Rivolta compares them with the
psorosperms of the liver of the rabbit, and points out that they
differ from these psorosperms in that they do not occur in the
epithelium, and that segmentation takes place in the body of the
host. He sums up his description by stating that there occur two
types of these oviform cells. In one type the contents consist of a
nucleus with four elongate corpuscles, while in the other there is a
large granular nucleus which at times 1s in process of segmentation.
In a later paper, Rivolta (1878) attempts to classify the psorosperms
and gregarines of animals. He names the oviform cells of the dog
and cat Cytospermium villorum intestinalium canis. He again
states that two types of this parasite occur. The first varied in
length from 8 to 12 microns, and had a breadth of 8 microns.
Within was a single elongate granular body like an embryo. After
a few days in water there developed three corpuscles and a granular
nucleus. The second type was larger, and varied in length from
12 to 16 microns, and had a breadth of about 12 microns. The
contents consisted of a single large granular mass which sometimes
showed signs of segmentation.

From the above summary of Rivolta’s descriptions it 1s clear that
the larger type 1s the oocyst and the smaller one the sporocyst. He
correctly observed the division of the granular mass into two
sporoblasts, but did not realise that each of these gave rise to one
of the smaller types which are sporocysts. It is evident that the
wall of the oocyst was not very resistant, and easily liberated the
sporocysts. In his earlier papers he correctly noted and figured
the four sporozoites and the residual body within the sporocysts. It

237

is evident that the infection was limited to the internal tissues of the
villi, and did not occur in the epithelium. ‘he size of the oocyst
was 12 to 10 microns by 12 microns, and that of the sporocyst 8 to
12 microns by 8 microns. The development was often completed
before the oocysts had left the tissues. Incompletely developed
sporocysts continued their development in water.

In his book, already noted above, Leuckart (1879, p. 282)
discusses the changes produced in the intestinal wall by coccidia
generally. He says he has seen these parasites in both dogs
and cats. In the latter animals he states that they occur in
the epithelium, where complete development takes place. In the
case of the dog, they were in the villi, and he evidently regarded
them as similar to the structures seen in this situation by Finck, but
is doubtful about those described by Rivolta. As regards the cat,
Leuckart is the only observer to refer to the complete development
of the oocyst in the epithelium. Finck and Rivolta, together with
Railliet and Lucet and Stiles, whose observations are considered
below, all state that this takes place in the deeper tissues of the villi.
Leuckart’s account is not always clear as to the actual animal he 1s
referring to, but the statement quoted definitely refers to the cat.
As will be explained below, the oocysts of coccidia which develop
in the epithelium do not commence to develop till they have left the
body, so that Leuckart’s statement is difficult to understand. It is
possible that oocysts of the larger forms in the epithelium might
develop in animals which had been dead for some considerable time,
or that Leuckart actually observed the oocysts of the small form in
an unusual situation in the epithelium. On the other hand, he may
have seen both a large and a small form in these animals, and
confused the two. It seems impossible to be certain of the form he
refers to in the cat, but his statements about the one in the dog are
much more precise.

The next observer to make a contribution to the subject from
personal cbservations was Grassi (1879), who gives a brief account
of a coccidium which he calls Coccidium Rivolta, from the intestine
of the cat. The oocyst is described as giving rise to two spores,
each of which contains four germs. In later papers (1882, 1883) he
gives under the same name a more detailed description. Ihe
oocyst is said to be elliptical in shape with one end more pointed

238

than the other. At the pointed end there could be detected a sort
of spiracle or micropyle. ‘The measurements of the oocyst are given
as 30°8 to 27 microns by 24 to 22 microns. W5thin it is a sphere
varying in diameter from 10 to 20 microns, with a central clear area
or nucleus. The sphere divides into two daughter spheres, each
having a diameter of 14°3 microns. [wo sporocysts result, within
which are found four embryos and a large residual body. It is
important to note that the parasite is described as occurring in the
epithelium of the intestine. The description 1s accompanied by
figures which illustrate clearly the structure of the oocysts. From
Grassi’s account there can be no doubt that he was dealing with a
coccidium which was entirely distinct from that described by Finck,
Virchow and Rivolta. As pointed out above, this distinction was
recognised by Perroncito (1882) and others.

Pachinger (1887) states that he had seen a sporozoon in the
oesophagus, stomach and whole length of the intestine of the
domestic cat, and that he had encountered a similar form in the
kidney of the dog. He says that it belonged to the monospore
coccidia with four sickle-shaped bodies. It is probable he was
observing the sporocysts of an /sospora of the cat, but there are no
means of identifying it with certainty, as no measurements were
given. The structures he records from the kidney of the dog are
quite unidentifiable.

Railliet and Lucet (1888) published an account of oviform bodies
which they found in the villi of a dog. ‘They noted their occurrence
in pairs, and remarked on their resemblance to the bodies described
by Virchow and Rivolta. On account of their association in pairs,
they hesitated to pronounce an opinion as to their coccidial nature.
After further study, Railliet and Lucet (1890) gave a brief but clear
description of these bodies as coccidia which they had observed in
the pole cat as well as the dog. In the dog the oocysts are said to
vary in length from 12 to 15 microns, and in breadth from 7 to
g microns. ‘The contents of each divide into two masses, and each
of these gives rise to four spores. The fully developed oocysts may
occur in the fresh villi, but usually complete development does not
take place till they have been in water for a few days. ‘The similar
form with oocyst, measuring 8 to 12 microns by 6 to 8 microns,
discovered in the pole cat (Mustela putorius) occurs in the deeper

tissues of the villus.

209

Stiles (1891) refers to the work of Railliet and Lucet, and says
that he has seen the cysts in the villi of dogs. He noted that each
might contain a single large mass of cytoplasm or two separate
masses suggesting a division into two of the large mass. Stiles
gives the name Coccidium bigeminum to this parasite. Raulliet and
Lucet (1891), 1n a further communication on the subject, accept the
name Coccidium bigeminum given by Stiles. ‘hey refer to the work
of Rivolta and Finck, and say there 1s no doubt that these observers
had studied the same organism. ‘They now describe three varieties
of the parasite as occurring in the dog, cat and pole cat, which they
regard as varieties of Coccidium bigeminum owing to differences
in the size of the oocysts :—

Coccidium bigeminum var. canis 12—15 X 7—Q microns.
Coccidium bigeminum var. cati 8—10X 7-9 ,,
Coccidium bigeminum var. putori 8-12 X 6—8 ,,

As pointed out by Wasielewski (1904) these variations in size are
insufficient to justify a separation of varieties on this basis alone.

The papers by Railliet and Lucet are not illustrated, but a figure
by Railliet appears in the English translation of Neumann’s work on
‘Animal Parasites’ (1892, p. 437). his figure again appears in
the second edition of the 77azté de Zoologie Médicale et Agricole
by Railliet (1895, p. 145). Stiles (1892) gave a fuller and illustrated
account of the Cocczdium bigeminum of the dog. He described the
development of the oocyst with the production of two sporoblasts
and two sporocysts, and the formation within each sporocyst of tour
sporozoites and a residual body. Ai figure of a section of the villus
shows the presence of oocysts containing the sporoblasts or
undeveloped sporocysts within the deep tissues of the villus. [he
size of the oocyst is given as 14 by 8 microns.

From the description of Railliet and Lucet, and Stiles, it is
evident that they were dealing with the coccidium seen by Finck,
Virchow and Rivolta in dogs and cats. ‘hese observations appear
to be the last ones which have been made on the small /sospora of
these animals.

Wasielewski (1904) gave a detailed account illustrated with
excellent microphotographs of the oocysts of a coccidium, called by
him Diplospora bigemina, which he had observed in cats that were
used for experiments on amoebic dysentery by Jiirgens. The
oocysts which he observed varied in size, and he gives a series of

240

measurements in microns as follows :—22 by 19, 25 by 20, 25 by 22,
35 by 23, 35 by 25, 35 by 27, 38 by 32, 40 by 28. He describes the
development of the oocyst in detail. The contents contract to form
a sphere, which has a diameter of 18 to 25 microns according to the
size of the cyst. Two daughter spheres are formed by division
of the large sphere, and these vary in diameter from 16 to 18 microns
in the larger oocysts and from 11 to 12 microns in the smaller ones.
The daughter spheres become sporocysts, within each of which are
developed four sporozoites 11 to 12 microns in length and a residual
body 6 to 8 microns in diameter. The earlier stages of the parasite
were found only in the epithelium of the small intestine and never
in the submucosa, so that Wasielewski considered that the statements
which had been made of a coccidium limited to the submucosa
required some qualification. Schizonts in the epithelium and motile
merozoites free in the lumen of the intestine were also seen.

This coccidium is clearly distinct from that studied by Finck,
Virchow, Rivolta, Railliet and Lucet, and Stiles. From the size
of the oocysts they appear to fall into two categories, as noted by
Reichenow (1921), the one with oocysts measuring 22-25 by 19-22
microns and the other with oocysts measuring 35-40 by 23-32
microns. ‘Those of the first category clearly correspond with the
parasite described by Grassi (1879, 1882, 1883). Wasielewski also
gave measurements of 18 by 25 microns for the oocysts and 11 by
15 microns for the sporocysts of a form seen by him in the dog.
He regarded it as Coccidinm bigeminum, but it corresponds exactly
with Grassi’s Coccidium Rivolta.

Basset (1909) without giving any description of the parasites,
discusses the pathogenic effect of coccidia, which he calls Didlospora
bigemina, in young dogs. He also records a round coccidium,
I4 microns in diameter, as occurring in dogs and ferrets, but there
is no evidence that these were actually coccidia, as no mention is
made of any development.

Swellengrebel (1914) gave a complete account of the development
of a coccidium of the cat under the name J/sospora bigemina,
which appears to be identical with the large form noted by
Wasielewski. He described for the first time the process of
schizogony in the epithelial cells of the small intestine, the evolution
of the macrogametocytes and microgametocytes, and formation and

241

development of the oocysts. The measurements of the oocyst are
given as 39 to 47 microns by 26 to 37 microns. The sporo-
cysts vary in length from 21 to 24 microns, and in breadth from
18 to 19 microns. Within the sporocyst there are formed four
sporozoites measuring 18 by 4 microns, and a large residual body.
Swellengrebel clearly states that the appearances are absolutely
unlike those figured by Stiles, but hesitates to establish a new
species.

Weidman (1915) described a coccidium, which he called
Coccidium bigeminum, in ‘swift foxes’ in the Western United
States. The oocysts varied from 25 to 40 microns in length by
25 to 30 microns in breadth. The sporocysts measured 16 to 20
by 14 to 18 microns. Owing to the difference in dimensions from
the form described by Railliet and Lucet, and Stiles, Weidman

oe ee]

canivecolis’’.’ He gives figures

suggests the ‘ new varietal name
of the oocyst containing two sporocysts, with four sporozoites and
a residual body. Mesnil (1916) states that Weidman regarded it as
a variety canivecolis of Isospora bigemina.

Wenyon and O’Connor (1917) found an /sospora of the cat very
common in Alexandria, and Dobell (1919) records a_ similar
experience in England in the case of cats used for experiments on
amoebic dysentery. In both these instances the oocysts were of the
large type. This was also the writer’s experience during experi-
ments on cats conducted in London in 1912.

Hall (1917) discovered a coccidium in dogs in Detroit. On account
of its large size, he thought it was different from /sosfora bigemina,
but later Hall and Wigdor (1918) concluded that it was a larger form
of the same parasite, and wrote of it as Dzplospora bigemina. The
oocysts measured 36 to 40 microns in length by 28 to 32 microns in
breadth. The sporocysts had a diameter of 10 to 20 microns, and the
sporozoites measured 12 by 4 microns. Oocysts of these dimensions
occurred in the majority of dogs, but in one animal a smaller strain
was seen, the oocysts measuring 20 by 18 microns and the sporocysts
12 by 11 microns, with sporozoites 10 microns in length by 3 microns
in breadth. ‘They state that this distinction in the size was quite
marked, and that it raises the question as to whether the small one
should be regarded as a variety or species. ‘They go on to say that
it is possible that there are several species.of Diflospora in the dog

242

characterised by considerable difference in size. The length of time
required for the development of the oocyst of the larger form was
two days when kept in 10 per cent. potassium bichromate solution.
Under other conditions, which they state more nearly resemble those
of nature, the time required may be two weeks or longer.

Reichenow (1921), referring to the /sospora of cats and dogs,
expresses it as his opinion that Wasielewski was probably dealing
with a mixed infection of two distinct coccidia in the cats he
examined. He also states that the form he had observed in the dog
in Germany differs from that in the cat, and resembles the one with
smaller oocysts studied by Wasielewski. For the oocysts of the dog
form he gives a length of 21 to 24 microns, and a breadth of 18 to
20 microns. ‘The sporocysts, which are oval in outline, measure
14 to 16 microns by 9 to 10 microns. No6ller (1921), in a brief
reference to the coccidium of dogs and cats, refers to the large and
small form in cats and the one in dogs. He has been able to infect
young dogs in series with the oocysts. No details of the dimensions
are given. Marotel (1922) studied the /sospora of the cat. His
measurements are as follows :—

Oocysts 45—48 X 34— 36 microns.
Sporocysts 22—24 X 17—19 microns.
Sporozoites 18 —20 X 4—5 microns, residual body in sporocyst
IO— [2 microns.
He proposes to call the coccidium /sospora cati. In order to
facilitate the following discussion, the various dimensions in microns
given by the above observers for the oocysts and sporocysts of the

dog and cat parasites are arranged in tabular form :—

Taste I.
. Oocyst Sporocyst
Finck (cat) a Ze an ee wae a 8—10x7—9
Virchow (dog) . = OY es ...| Like those described by Finck sat
Rivolta (dog and ‘cat) ts vs ape EE Oe See 8 — 12 x 8
Grassi (cat) “te aes woo|. 27 — 30°38 K 22 — 24 14°3
Railliet and Lucet (dog) te =f abd pe t2 =I X17) 9:
— (cat) ous 5 ..| 8—10xX7—9
(pole cat)... A ...| 8—12 x 6—8
Stiles (dog) 4 ae my, a »..| 13°6 — 15°99 X 7°99 — 9°9 a
Wasielewski (cat) = 9 vs oe | 35 — 40 X 23 — 32 16 — 18
(cat) vst fs des sadly 22 25. X10 22 10 — 12
(dog) a A ey oer Oo eeees II — 15
Swellengrebel (cat) ... dé. oh .o| 39 — 47 X 26 — 37 21 — 24 X 18 — 19
Hall and Wigdor (dog)... a a .--| 36 — 40 X 28 — 32 Io — 20
(dog)... Ls ass il .30 348 (2¥K a2

Reichenow (dog) — ‘x ah .-.| 21 — 24 X 18 — 20 14 — 16 X 9— 10
Marotel (cat) ... aie ees th wes) 45 — 48 X 34 — 36 22 — 24 X 17 — 19

243

From the above table it will readily be seen that the oocysts
described fall into three groups.

(1) There are the small forms described by Finck, Rivolta,
Railliet and Lucet, and Stiles. Finck did not state the actual
measurements of the oocysts, but from the size given for the sporo-
cysts and the fact that in his description he says that two of these
sometimes occur together enclosed by a common membrane, it is safe
to assume that the oocyst would have dimensions similar to those
described by Rivolta, Railliet and Lucet, and Stiles. The forms
seen by Virchow are evidently similar, for he says they occur in the
interior of the villi of dogs, are relatively small and regularly
arranged in pairs enclosed by a thick, double contoured membrane.
It is probable also that those described by Leuckart are of the
Same type. ;

(2) The second type has an oocyst of intermediate size. This
was first seen by Grassi in the cat, later by Wasielewski in the cat
and dog, by Hall and Wigdor in the dog, by Reichenow in the same
animal, and possibly by Ndller in the cat and dog.

(3) The third type has an oocyst of much larger size. This was
first definitely described by Wasielewski and Swellengrebel in the
cat, and was seen by Wenyon and O’Connor, Dobell, Hall and
Wigdor, and Marotel.

That these three types represent distinct species seems clear from
the above records, and from observations to be recorded in this
paper. In a recent study of English cats, the oocysts which
occurred in the faeces were uniformly of large size, while those which
were found in dogs’ faeces were of the intermediate type. In one
instance only was an infection of the cat with the small type seen. In
this case the large form occurred also and it was clearly evident that
the small one was limited to the deeper tissues of the villus, while
the large one deveioped in the epithelium. Furthermore, develop-
ment of many of the small oocysts was completed in the tissues of the
villi, while those of the large form did not take place for some days
after it had left the body. That the oocysts of the smallest form
sometimes escape in the faeces in the undeveloped condition 1s
demonstrated by an observation which has just been made by
Mr. Leslie Sheather, of the Royal Veterinary College, with whom
the writer has discussed his investigations on coccidiosis of dogs and

244

cats. By a process of concentration employed for the detection of
worms’ eggs in faeces, Mr. Leslie Sheather discovered that one dog
was infected with the small form and another with that of inter-
mediate size. The small oocysts measured about 12 microns in
longest diameter, and like those of intermediate size were in the
undeveloped condition. They proceeded to development when kept
outside the body.

NOMENCLATURE

As regards the nomenclature of these parasites, there appears to
be no great difficulty, though the name Coccidium bigeminum
Stiles, 1891, has been employed indiscriminately for all three
forms. Apart from Rivolta’s name Cytospermium villorum
wntestinalium canis which he proposed in 1878, Grassi’s name
Coccidium Rivolla (1879) 1s the first one to be given to any one of
these Coccidia. As pointed out above, Grassi was dealing with the
oocysts of intermediate size in the cat, and, assuming that this form
is the same as that of corresponding size from the dog, his name has
priority. The name of this coccidium 1s, therefore, /sosfora rivolta
(Grassi, 1879). For the small form in the dog and cat the correct
name 1s /sospora bigemina (Stiles, 1891). This leaves the large
form in the dog and cat still unnamed, for the name /sospora cati
suggested by Marotel (1922) cannot stand, as Railliet and Lucet
(1891) employed the name Coccidium bigeminum var. cati for the
small form in the cat, which if recognised as a distinct species from
that in the dog, would become /sospora catz. For the large
species the name /sospora felis is suggested. There are thus to be
distinguished in dog and cat three species of /sospora :—

Isospora bigemina (Stiles, 1891).
Isospora rivolta (Grassi, 1879).
Isospora felis n. sp.

It is assumed that these different parasites are able to infect
both dogs and cats, but it 1s possible that each animal has its own
species. This can only be determined by more detailed observation
and cross-infection experiments with clean animals. Railliet and
Lucet (1891) have stated that the small forms in the cat, dog and
pole cat are varieties of /sospora bigemina, while Weidman (1915) has
made a similar suggestion for the large coccidium described by him

245

in the fox. His name was not properly proposed, as he merely says
he advances a new varietal name ‘ canivecolis.’ Mesnil (1916), in
a summary of Weidman’s paper, writes the specific name canivecolis,
while Hall and Wigdor (1918) give it in full C. digeminum
canivecolis. his parasite, which is certainly not a variety of
Isospora bigemina, may be identical with /sospora felis, but on the
other hand it may be distinct. In the latter case the name /sospora
canivecolis would be correct.

There exists also in dogs in England ah intestinal Eimeria as
recorded by Brown and Stammers (1922). For this parasite the
name Limeria cants 1s proposed.

Though Grassi (1879) proposed the name Coccidium Rivolta for
the parasite he found in the cat, this name has been modified by
several observers, in spite of the fact that Grassi repeated the name
in his later papers (1882, 1883). Dobell (1919) in discussing this
question, says that it 1s his view that Grassi’s name should be
changed by putting ‘zzvolta’ in the genitive, in which case the

name would be /sespora rivoltae. He thinks that a form such as

“rvivoltat’ is objectionable. There seems, however, to be no real
reason why the name should he changed at all, and to keep
it in the form proposed by Grassi is in accordance with Rules of
Nomenclature. Both the changes discussed by Dobell have,
however, been previously made. Thus in the English translation
of Leuckart’s work (1886) there appears a note on page 221 initialed
by the author (R. L.) in which the name Cocctdium Rivoltae, Grassi
is used for the first time. Raiulliet (1895, p. 146) uses the name
Coccidium bigeminum Stiles, 1891, for the small coccidium of the
cat, dog and pole cat, and the name Coccidium (?) Rivoltat Grassi,
1881, for the form of intermediate size seen by Grassi. Neveu-
Lemaire (1912) employs the name Lzmerza Rivaltai Grassi, 1881,
for the latter form, while Brumpt (1922), 1n the latest edition of his
Précis de Parasitologie, uses the name /sospora Rivoltat Grassi, for
all these parasites.

Several observers, including Wasielewski (1904), Martin (1909),
Guiart (1910) and Hall and Wigdor (1918), place these parasites in
the genus Diplospora, which, however, is generally recognised as a
synonym of /sospora.

For convenience of reference, the following list of names

246

which have been employed for the /sospora of cats and dogs is
appended : —

Finck (1854). Corpuscules géminées.

Vulpian (1858). Corps oviformes.

Ercolani (1859). ? (Quoted by
Rivolta and Perroncito.)

Virchow (1860). Psorospermien.

Leuckart (1860). Psorospermien.

Davaine (1860). Corpuscules géminées.

Leuckart (1863). Psorospermien.

Leuckart (1866). Psorospermien.

Eimer (1870). Psorospermien.

Ziirn (1874). Psorospermien.

Rivolta (1874). Cellule oviforme.

Rivolta (1877). Cellule oviforme.

Davaine (1877). Corpuscules géminés.

Rivolta (1878). Cytospermium villorum
intestinalium canis.

Leuckart (1879).

Grassi (1879). Coccidium Rivolta.

Grassi (1882). Coccidium Rivolta.

Biitschli (1882) Coccidium Rivolta
Grassi.

Perroncito (1882).
Coccidium Rivolta.
Cytospermium villorum

alium canis.

Braun (1883). Cocctdium perforans.

Grassi (1883). Coccidium Rivolta.

Balbiani (1884). Coccidium perforans.

Leuckart (1886). Coccidium Rivoltae,
Grassi.

Railliet (1886).
Grassi.

Pachinger (1887). Sporozoon.

Neumann (1888). Coccidium perforans.

Railliet and Lucet (1888). Corps
oviformes.

Ziirn (1889).
Leuck.

Blanchard (1889). Coccidium Rivolta
Grassi, 1881.

Pfeiffer, L. (1890). Coccidien.

Railliet and Lucet (1890). Coccidies.

Stiles (1891). Coccidium bigeminum.

Pfeiffer, L. (1891). Coccidien.

Railliet and Lucet (1891). Coccidium
bigeminum vars. canis, cati, putori.

Stiles (1892). Coccidium bigeminum
Stiles, 1891.

intestin-

Coccidium Rivolta

Coccidium oviforme

Coccidium perforans.

Neumann (1892).
Coccidium bigeminum.
Coccidium perforans.
Coccidium Rivolta Grassi.
Mosler and Peiper (1894). Coccidien.
Railliet (1895).
Coccidium bigeminum Stiles, 1891.
Coccidium(?) Rivoltat Grassi, 1881.
Braun (1895). Coccidium bigeminum
Stiles 1891.
Moniez (1896). Coccidium bigeminum
Stiles (1891).
Blanchard (1896). Cocctdium bigemt-
num Wardel Stiles, 1891.
Labbé (1896). Coccidium bigeminum
Stiles.
Wasiclewski (1896).
Coccidium bigeminum Stiles.
Coccidium spec. inc. Rivolta Grassi.
Labbé (1899). Coccidium bigeminum
Stiles.
Blanchard (1900). Coccidium bigemt-
num Wardell Stiles, 1891.
Neveu-Lemaire (1901). Coccidium
bigeminum Wardell Stiles, 1891.
Doflein (1901). Coccidium bigeminum
Stiles.
Perroncito (1901).

Coccidium bigeminum Wardell
Stiles, 1891.
Coccidium Rivolta.
Neveu-Lemaire (1902). Coccidium

bigeminum Wardell Stiles, 1891.
Neveu-Lemaire (1903). Coccidium
bigeminum Wardell Stiles, 1891.
Braun (1903). Coccidium bigeminum

Stiles, 1891.

Minchin (1903). Coccidium bigemt-
num vars. canis, catt, putort Railliet
et Lucet.

Wasiclewski
bigemina.

Neumann (1905).

Coccidium bigeminum.
C. Rivoltae (Grassi).

Guiart and Grimbert (1906). Cocci-
dium bigeminum Stiles.

Lihe(1906). [sospora bigemina (Stiles).

Braun (1906). Coccidium bigeminum
Stiles, 1891.

(1904). Diplospora

247

Neveu-Lemaire (1906). Coccidium Brumpt (1913). Coccidium bigeminum

bigeminum Wardel Stiles, 1891. (Wardel Stiles, 1891).
Braun (1908). Jsospora bigemina Swellengrebel (1914). Isospora

(Stiles) 1891. bigemina (Stiles).
Neveu-Lemaire (1908). Coccidium Braun and Seifert (1915). Isospora

bigeminum Wardel Stiles, 1891. bigemina (Stiles) 1891.
Basset (1909). Diplospora bigemina. Doflein (1916). Isospora bigemina
Guiart and Grimbert (1908). Cocct- (Stiles).

dium bigeminum Stiles. Fantham (1916). Isospora bigemina,
Braun and Liihe (1gog). Isospora Stiles, 1891.

bigemina (Stiles). Wenyon and O’Connor (1917). Isos-
Martin (1909). Dziplospora bigemina pora of cats.

Stiles. Hall and Wigdor (1918). Dzplospora
Doflein (1909). Isospora bigemina bigemina.

(Stiles). Dobell (1919).
Braun and Liihe (1910). Isospora Isospora bigemina Stiles.

bigemina (Stiles). Isospora rivoltae Grassi (1879).
Brumpt (1910). Coccidium bigeminum Reichenow (1921). Lsospora bigemina

Wardel Stiles, 1891. (Stiles).
Guiart (1910). Dzplospora bigemina. Dobell and O’Connor (1921). Lsospora
Doflein (1911). JLsospora bigemina rivoltae Grassi.

(Stiles). Noller (1921). Lsospora bigemina.
Fiebiger (1912). Isospora bigemina Mayer (1922). Coccidien ?

Stiles. Brumpt (1922). Isospora Rivoltat

tv Neveu-Lemaire (1912). Eimeria Grassi.
Rivoltat Grassi, 1881. Marotel (1922). Isospora catt.

Jollos (1913). Isospora bigemina.

DESCRIPTION OF THE COCCIDIA OF CATS AND DOGS

During the course of certain observations on the faeces of dogs,
the results of which have been published by Brown and Stammers
(1922), it became evident that dogs were sometimes infected with a
species of /“zmerza in addition to the commonly recognised /sospora.
The oocysts of the latter parasite agreed, as regards dimensions, with
those given by Grassi (1879, 1882, 1883) for the /sospora of the cat,
by Wasielewski (1904) for the /sospora of the dog and small form
in the cat, and by Reichenow (1921) for one in the dog, and were
constantly smaller than the oocysts of the /sospora of the cat which
was under observation at the same time, so that it seems highly
probable that the common /sospora of dogs and cats belong to two
distinct species. The oocysts of the Ezmeria of the dog varied
considerably in size, some of them being as large as those of the
large /sospora found in the cats, while others were smaller even than
those of the /sospfora seen in the dogs. They differed in appearance
from the oocysts of the /sospora of the same animal and showed a

248

much greater range in size, but it was only after material had been
kept till development of the oocyst had completed itself that it was
definitely recognised as an Ezmeria. It is possible that this Ezmeria
has been seen before and regarded as an /sospora, but of this there
is no evidence.

In the case of the /sospora of the cat the oocysts examined by
the writer have been constantly of large size, except in one instance
when very much smaller ones were also present. It was found by
examination of the small intestine of this cat that the large oocysts
were derived from an /sospora (Isospora felts) which was undergoing
development in the epithelium of the intestinal villi, while the very
much smaller ones belonged to another /sospora (Isospora bigemina)
which was parasitic only in the deeper tissues of the villi.
Furthermore, the oocysts of the latter form completed their develop-
ment in the tissues, whereas those of the large form in the epithelium
were in the usual undeveloped condition. The faeces of this cat had
been examined on several occasions in connection with experiments
with Extamoeba histolytica, but the only oocysts noted in the faeces
were the large undeveloped ones of /sospora felis. The oocysts of
Tsospora bigemina were not seen in the faeces, and if they had been
present to any extent they could not have escaped recognition. They
were first detected when a scraping of the wall of the small intestine
was made with a view to finding amoebae which had been seen in
this situation in another cat with amoebic dysentery. It seems clear
that the oocysts of the small form do not escape into the faeces so
regularly as do those of the large one which develops in the
epithelium. There is no doubt that there were two distinct species
of /sospora in this cat.

A detailed study of the development of /sospora felis and
Isospora bigemina as they occurred in the tissues of cats was
undertaken, but /sospora rivolta of the dog was only investigated in

the oocyst stages which occurred in the faeces.

ISOSPORA FELIS 2. sp.

The only complete account of the development of this common
coccidium of the cat is that of Swellengrebel (1914), though
Wasielewski (1904) had described the development of the oocyst
and had seen other stages in the epithelium. In its main outlines

249

Swellengrebel’s description 1s correct, but the growth of the micro-
gametocyte was not fully traced. The supposed parthenogenesis of
the macrogametocyte 1s capable of another interpretation, while the
account of the changes undergone by the nuclei requires revision.
It seems, therefore, desirable to redescribe the life-history as it has
been studied in sections of the epithelium of the small intestine
of cats.

Schizont. The smallest forms which can be found in the
epithelial cells are only 5 microns in length (Plate IX, fig. 1). These
are curved and somewhat sickle-shaped bodies which are pointed
anteriorly and rounded posteriorly. They lic in vacuoles in the
cells, and are attached to the cytoplasm of the cell by the pointed
extremity. The nucleus is spherical and has a definite membrane.
Within the nucleus is a body which, in staining reactions, does not
appear to be rich in chromatin. It is usually applied to the nuclear
membrane. In addition the nucleus contains a granular material
which is probably chromatic in nature. Whether the large body
should be regarded as a karyosome depends on the definition of this
term. It does not stain intensely with Mayer’s haemalum, has
the appearance of plastin material rather than chromatin, and in this
respect resembles a nucleolus rather than a karyosome. Growth of
the parasite takes place till it has a length of about 10 microns
and a diameter at its thickest part of about 5 microns (Pl. IX,
figs. 2 and 3’. Though plumper than the youngest forms, it still
retains its elongate gregariniform character. While still in this
condition nuclear division commences (PI. IX, fig. 4) by division of
the karyosome. The daughter karyosomes take up positions at the
end of the now elongated nuclear membrane as two polar caps while
a definite equatorial plate of small chromosomes is formed (PI. IX,
fig. 5). Two daughter plates are formed and the first nuclear
division is completed by division of the membrane (PI. IX, figs. 6-8).
The two daughter nuclei have the same structure as that of the
original nucleus. The second nuclear division takes place in a
similar manner, as does also the third, though the karyosomes as a
tule become smaller with each division (Pl. IX, figs. 9-12). When
eight nuclei are present, the parasite has become more definitely
ovoid in shape, and eight merozoites are formed by a budding
process which leaves a definite residual body. Eight appears to

250 ‘

be the usual number for the merozoites, for the vast majority of
schizonts which have been seen are of this type. The size of the
merozoites, however, varies considerably even when only eight are
present (PI. IX, figs. 14 and 15). It seems possible that the small
forms are destined to develop again into schizonts and the larger
ones into gametocytes, but no definite proof of this could be
obtained. Occasionally a smaller number of merozoites appeared
to be formed (Pl. 1X, fig. 16), but in such cases it is possible that
the appearance was due to multiple infection of a cell by merozoites
after schizogony had occurred, or to the fact that all the merozoites
resulting from schizogony had not escaped from the cell. In several
instances the occurrence of two merozoites ina single vacuole was
undoubtedly due to two merozoites having invaded the same cell
simultaneously. On the other hand, a large number of merozoites
is sometimes formed, as pointed out by Swellengrebel (Pl. IX,
figs. 17 and 18). In several instances as many as sixteen occurred,
while a larger number was once seen. These forms, however,
occurred rarely in the material examined, and, as stated above, the
great majority of schizonts produced only eight merozoites.

It should be pointed out that the schizonts tend to stain very
deeply, even with very dilute Mayer’s haemalum, which proved to
be the most satisfactory stain for these forms, so that unless thin
sections are examined there may be considerable difficulty in making
out the details of the nuclear divisions.

During growth the schizont 1s closely applied to the nucleus of
the host cell, which becomes definitely altered in character. -

Microgametocyte. The microgametocyte possibly commences as
one of the larger merozoites (Pl. IX, fig. 19). Like the schizont,
it retains, for a considerable period of its growth, its gregariniform
character. When it has a length of about 12 microns (Pl. IX,
fig. 20) the first nuclear division takes place. ‘This is very similar
in character to that of the schizont. ‘The karyosome is present and
divides in the same manner by dumb-bell constriction, while there
is evidence that chromosomes are also formed (Pl. [X, figs. 20-22).
Repeated nuclear divisions of the same type take place while the
microgametocyte increases steadily in size. It finally loses its
gregariniform character and becomes irregular in shape till it has a
length of about 20 microns (Pl. IX, figs. 23-28). The increase in

251

bulk up to this stage has been relatively enormous. The details of
the nuclear divisions are difficult to follow owing to the marked
affinity the cytoplasm has for stains. This obscures details to such
an extent that it 1s very difficult to detect the arrangement of the
chromatin during the divisions of the nucleus.

After this a change takes place. The cytoplasm ceases to stain
intensely, the chromatin material in the nucleus becomes much more
definite and the karyosome decreases in size. Nuclear divisions
continue, and these are definitely mitotic in character (Pl. X,
figs. 1-3). The chromosome number has not been counted with
accuracy, but it appears to be somewhere within the limits of
8andi2. It appears that the nuclear membrane persists throughout
nuclear division. The cytoplasm becomes fissured in various ways
and loses still more its affinity for stains. Finally, when nuclear
division 1s complete, the microgametocyte contains a large number
of nucle: which have definite nuclear membranes within which are
irregular masses of chromatin (Pl. X, fig. 4). The karyosome,
which had decreased in size during the later divisions, 1s no longer
clearly visible, but it seems probable that it is still present, for in
the later divisions of the nucleus it is often possible to detect a small
granule at each end of the mitotic figure. These two granules may
be united by a fibre, so that the appearance of a minute karyosome
dividing by elongation and constriction 1s produced. The nucle
then shrink, and become compact, deeply staining masses of
chromatin (Pl. X, fig. 5). |

Formation of microgametes commences by the outgrowth from
the nucleus of a short process (Pl. X, fig. 6). The whole nucleus
then elongates (Pl. X, fig. 7), and it seems probable that the
short process represents the anterior end of the microgamete. The
short curved masses then become more elongate, and fine tapering
microgametes about 5 microns in length are formed (PI. XI, fig. 1).
The cytoplasm of the microgametocyte either collects into a single
large residual body on the surface of which the microgametes he,
or it breaks up into several separate masses. A certain number of
deeply staining granules remain in the residual body. The
individual microgamete is pointed anteriorly and fine and tapering
posteriorly. Sometimes there appeared to be a deeply staining
granule near the anterior end of the microgamete. It is possible

252

that this granule functions as a blepharoblast from which the
two flagella which Swellengrebel demonstrated arise. It seems
probable that this granule is the karyosome or, as some would
term it, the centriole which could be detected during the later
divisions of the nuclei of the microgametocyte. When develop-
ment of the microgametocyte 1s completed it may have a length

_ of nearly 50 microns and measure over 30 microns in the two

other diameters, so that it appears in many sections of a series.
Well over two thousand microgametes may be formed by each micro-
gametocyte. Swellengrebel was unable to trace the complete
development of the microgametocyte, but it appears from his figures
that some of the forms which he regarded as developmental stages
of schizonts are really microgametocytes.

Macrogametocyte. I\t is assumed that the macrogametocyte
commences as one of the larger merozoites (Pl. XI, fig. 2). At this
early stage it has been impossible to differentiate between the young
stages of either the muicrogametocytes, macrogametocytes or
schizonts. The macrogametocyte can, however, be recognised at
later stages owing to the fact that it has increased in size without
nuclear division. It retains its gregariniform character, and 1s
attached to the surface of the vacuole in the cell by its pointed
extremity. The attachment is frequently on the nucleus, which in
some cases 1s drawn into the vacuole (Pl. XI, figs. 3 and 4). On
several occasions what appears to be a definite organ of attachment
was seen (Pl. XI, fig. 5). Sometimes there 1s an appearance of a
terminal sucker which has drawn into it a small pedicle of the
cytoplasm of the cell. Even when the macrogametocyte reaches a
large size the gregariniform shape 1s retained, so that the parasite
may become doubled to accommodate itself to the space at its
disposal (Pl. XI, figs. 6-8). During the stages of growth repre-
sented by Pl. XI, figs. 2-8 the cytoplasm stains deeply, with a
tendency towards the accumulation of more intensely staining
material round the nucleus in the later stages. The nucleus has
increased considerably in size and possesses a large karyosome which
has little affinity for stains. A change now takes place in the
staining reactions. The deeply staining material round the nucleus
increases in amount and there appears in the cytoplasm a number
of deeply staining irregular bodies, while the cytoplasm itself
becomes filled with vacuoles containing a clear refractile substance

253

(Pl. XI, fig. 9). The cytoplasm generally has less affinity for stains
than it had previously, and it seems as if the substance which caused
the cytoplasm to stain deeply in the earlier stages has now
become aggregated in the irregular masses. The latter eventually
disappear, leaving a clear cytoplasm filled with refractile globules
(Pl. XI, figs. to and 11; Pl. XI, fig. 1). Finally the oocyst is
secreted round the macrogametocyte. It does not become thick or
resistant till it leaves the cell, for in fixed tissues the oocysts within
the cells are permeable to fixatives and show no signs of the shrinkage
and lack of proper fixation which is characteristic of those which are
free in the lumen of the intestine.

During the growth of the macrogametocyte it is frequently noted
that a granular substance accumulates in the vacuole between the
parasite and the wall of the vacuole. This material, which often
stains brilliantly with eosin, causes indentations in the macro-
gametocyte in various places (Pl. XI, fig. 11). Similar accumula-
tions sometimes occur in the case of the microgametocytes (Pl. X,
ey yy!

Swellengrebel described a process of parthenogenesis of the
macrogametocyte. Nothing comparable with this has been seen
during the present investigations, and, judging from his figures,
it seems that the stages he figures, in which definite nuclei are not
present, are drawn from sections of macrogametocytes which did
not include the nucleus but showed the deeply staining material
which occurs around it (Text-fig. 2, p. 259). The large macro-
gametocytes naturally occur in several sections of a series, and the
nucleus may only be found in one of these. In the sections on
either side of this one the macrogametocytes will have the appearance
of the forms figured by Swellengrebel as illustrating his process of
parthenogenesis.

The foregoing description of the development of /sospora felis
in the intestinal epithelium of the cat is of interest from several
points of view. In the first place it 1s of importance to note that
the parasite is limited to the epithelial cells. In no case has it been
seen in the sub-epithelial tissues. The infection, moreover, appears
to be confined almost entirely to the epithelium near the distal ends
of the villi, there being little tendency for it to spread towards their
bases.

During the growth of the young forms of the schizont and

254

gametocyte the parasite retains its gregariniform character to a
relatively late stage. In this respect /sosfora felis differs from many
coccidia, which quickly assume the spherical form when growth
commences. The fixation of the growing forms to the surfaces of
the vacuoles by the pointed end and the development of what
appears to be a definite organ of fixation still further increases the
resemblance to certain gregarines, such as those of the genus
Lankesteria.

The development of the microgametocyte merits special attention
from the point of view of the behaviour of its nucleus. Schaudinn
(1900), in his description of Fimeria schubergi, stated that the
nucleus of the microgametocyte broke up into a chromidium, the
granules of which collected in the form of a number of nuclei on the
surface. A similar process was described by him (1902) for
Cyclospora caryolytica, and again by Schaudinn and _ Siedlecki
(1897) in the case of Ezmeria lacazez. The majority of observers
who have described the development of the microgametocytes of
coccidia have followed Schaudinn in supposing that the numerous
nuclei are formed from the chromidium into which the single nucleus
breaks up. It was shown by Schellack (1912, 1913) and by
Schellack and Reichenow (1913, 1915) for a number of coccidia,
including the forms with which Schaudinn himself worked, that the
latter’s statements were incorrect, and that the nuclei of the mature
microgametocyte resulted from repeated nuclear divisions from the
original nucleus. A similar process had been described by
Wasielewski (1904) for /sospora lacazei of birds, by Stevenson (1911)
in the case of the Ezmerza of the goat, by Léger and Duboscq (1910)
for Selenococcidium intermedium, and by Siedlecki (1899) for
Adelea ovata. It is very doubtful, therefore, if the microgamete
nuclei are ever formed from chromidium, as Schaudinn maintained.
It seems far more probable that in all coccidia they result from
repeated nuclear divisions, as described above for /sospora felis.

The structure which has beem called the karyosome is present in
all the stages of schizogony and in the merozoites. It occurs during
the early nuclear division stages of the microgametocyte, but in the
later stages is represented by a minute granule. Whether this is to
be regarded as a centrosome or centriole is a difficult question to
decide. It certainly occupies the position in mitotic division that

ae - a

255

a centrosome would occupy, and furthermore, it is probably this
granule which occurs at the anterior end of the microgamete, and from
it the flagella may originate. The karyosome is constantly present
during the growth of the macrogametocyte, though it usually becomes
smaller towards its maturity. Whether it disappears before fertilisa-
tion takes place has not been definitely determined, but it is certainly
present during the nuclear division of the zygote and sporoblast.
There was no indication that the karyosome was discharged from
the nucleus prior to fertilisation. Though the latter process was not
actually observed in stained preparations, in a few cases the nucleus
of the fully grown macrogametocyte was elongated. It seems
probable that this was an elongation preparatory to fertilisation, and
if so it is worthy of note that the karyosome was still present in the
nucleus.

Oocyst. As regards the oocysts themselves (Pl. XII, figs. 12-15)
the measurements of a large number showed that they vary in length
from 39 to 48 microns and in breadth from 26 to 37 microns, the
majority measuring about 45 by 33 microns. These figures are
practically identical with those given by Swellengrebel. Wasielewsk,
however, saw smaller forms in the cat, his measurements being
22 to 40 by 19 to 28. It seems possible that cats may be infected
with both J/sospora felis and J/sospora rivolta, in which case
Wasielewsk1’s figures would cover a mixed infection with these two
forms. Grassi appears to have been dealing with a pure infection
of /sospora rivolta in the cat.

The development of the living oocyst of /sospora felis has been
followed by Wasielewski, Swellengrebel and others, and there 1s
little to add to their descriptions. Owing to the impermeable nature
of the oocyst wall, it is difficult to obtain satisfactorily fixed
preparations of the nuclei during its development. A certain
number of preparations was, however, obtained in the following
manner. Small quantities of the material containing oocysts in
various stages of development were crushed between a slide and
cover-glass in order to rupture the cysts, and films fixed in
Schaudinn’s fluid and stained with iron haematoxylin were made in
the usual manner. ‘There was thus obtained a number of stained
preparations of the different stages.

The nucleus of the zygote (Pl. XII, fig. 2) has very much the

256

same appearance as that of the macrogametocytes in the tissues.
A karyosome is still present, though it appears to be smaller. The
same type of nucleus occurs in other stages, including those of the
sporoblasts (Pl. XII, figs. 3-7), but in these the karyosome has -
increased relatively in size. A few nuclear divisions were seen, but
these were not sufficiently numerous for many details to be made out.
The stages which were seen resembled those which occur in the
early stages of development of the microgametocyte, except that the
karyosomes are smaller. The nuclei m various stages of develop-
ment of the oocyst are depicted in Pl. XII, figs. 2-10.

The zygote nucleus (Pl. XII, fig. 2) 1s a spherical body consisting
of a definite membrane, within which a number of fine granules and
one larger mass—the karyosome—occur. Whether the karyosome
is present in the earliest stage of the zygote nucleus could not be
determined, as stained preparations of the fertilisation process were
not seen. Satisfactory pictures of the first nuclear division were not
observed, so that no statement can be made regarding a possible
reduction in the number of the chromosomes. ‘The two nuclei of the
binucleate stage are shown at Pl. XII, fig. 3. Both nuclei are
decolorized, and the small granule at the centre of the karyosome is
well seen. The single nuclei of the two sporoblasts have the same
structure. The first division in the sporoblast is shown at Pl. XII,
fig. 4. The daughter karyosomes occupy the poles of the spindle,
while daughter plates of chromosomes are also present. The nuclei
of the binucleate stage of the sporoblast are shown at Pl. XII, fig. 5,
and here again the nuclei are of the same type. The second nuclear
division in the sporoblast shows two spindles with the karyosomes
at the poles of the spindle, and definite equatorial plates (Pl. XII,
fig. 6). The resulting four nuclei, with somewhat deeply stained
karyosomes, are shown at Pl. XII, fig. 7.

Good preparations of sporozoites were fairly numerous. These
measured from 10 to 15 microns in length (Pl. XII, figs. 8-11), being
smaller after fixation than in the living condition. In some a large
vacuole occurs near one end. ‘This is evidently the position of the
refractile body often seen in the living sporozoites (Pl. XII, fig. 15).
The nucleus was spherical and contained a relatively large
karyosome. In specimens from which the stain had been sufficiently
extracted (Pl. XII, fig. 8) the karyosome was pale, and at its centre

257
was a small deeply staining granule. It thus appears that the
karyosome is present in all stages of the nuclei during sporogony,
though varying considerably in size.

The sporozoites appear to be budded off in pairs from the ends

of the sporoblast. Two buds appear at each end, and these grow
into elongate finger-like processes into which the nuclei enter.
During their growth they turn over the surface of the residual body
and le between it and the wall of the sporocyst.
» An important point to note is that the oocyst commences to form
as a thin membrane while the macrogametocyte is still within the
epithelium, but it does not become a resistant structure till the
macrogametocyte has left the cell for the lumen of the intestine. In
no case was there any indication that the further development of the
contents took place, either in the cells or in the lumen of the intestine.
Retraction of the zygote and division of the latter into two sporo-
blasts, which are the first steps in the development after the oocysts
leave the body, were never noted in the case of oocysts within the
epithelium or in the lumen of the intestine. It follows, therefore,
that whenever observers have described the occurrence of paired
bodies in the intestine wall they cannot have been referring to the
oocysts of /sospora felis.

ISOSPORA BIGEMINA (STILES, 1901)

This coccidium was discovered in one cat which had _ been
employed for experiments with Axtamoeba histolytica. The cat died
during the night, but at the autopsy next morning it was still warm
and perfectly fresh and the amoebae active and in healthy condition.
The cat had evidently been dead only a short time. It 1s important
to note this fact, for many of the oocysts of /sospora bigemina
which occurred in the submucosa were fully developed. It seems
hardly possible that they could have completed their development
in the short time following the death of the cat. This 1s all the more
probable in view of the fact that oocysts of /sospora felis which were
also present in the intestine were quite unchanged. It can safely
be assumed, therefore, that the appearance of /sospora felis in the
tissues and in the intestine were those which occurred during life.
This accords with the descriptions which have been given by Finck,
Virchow, Leuckart, Railliet and Lucet, and Stiles.

258

The sporocysts of this coccidium were first seen in scrapings of
the wall of the small intestine after the death of the animal. It was
at first thought that they were fully developed sporocysts of
Isospora felis, but their small size was against this view. Further
examination showed that they really occurred in pairs enclosed in
an oocyst which was easily ruptured between the slide and cover-
glass. The sporocysts had fairly thick, double-contoured walls,
and contained four sporozoites and usually a residual body. ‘The
oocyst wall enclosing them was of a more delicate nature, and was
closely wrapped round the two sporocysts. There was no indication
of a micropyle in the oocyst.

Sections of the small intestine showed that the parasite did not
occur in the epithelium, but was limited entirely to the sub-epithelial
tissues of the villi, especially near their distal ends, some of which
were swollen and packed with oocysts in various stages of develop-
ment. The epithelium contained /sospfora felis, which on account
of its large size contrasted very markedly with the much smaller form
in the tissues. Text-figure 2 1s from a drawing of a transverse
section of a villus, and shows two macrogametocytes, one with a
nucleus and the other with the central granular mass to the side of
the nucleus, a fully developed microgametocyte with microgametes,
and a young micro- or macrogametocyte of /sospora felis in the
epithelium, and six fully developed oocysts of /sospora bigemina in
the sub-epithelial tissues.

The earliest stages of /sospora bigemina are seen as minute
spherical bodies enclosed in vacuoles in the cytoplasm of mononuclear
cells (Pl. XIII, fig. 1). Whether these are endothelial cells or not
has not been determined. No endothelial cells which were evidently
on the walls of blood vessels were found infected. These young
forms are barely 2 microns in diameter. They grow into schizonts
which are 5 to 6 microns in diameter, and produce about twelve
merozoites. Owing to their small size, it is exceedingly difficult to
follow the development in the ‘sections (Pl. XIII, figs. 2-4). The
microgametocytes have not been definitely identified, though several
structures have been seen which are possibly of this nature. One
of these has been drawn (Pl. XIII, fig. 5), and it would seem not
improbable that the minute curved bodies are microgametes
surrounding a residual mass of cytoplasm. The macrogametocyte

259

develops into an ovoid body ro to 12 microns in length (Pl. XIII,
figs. 6and 7). It becomes enclosed in an oocyst (Plate XIII, fig. 8),
within which it divides into two sporoblasts, which in their turn
form sporocysts the walls of which are thicker than that of the oocyst.
Within each sporocyst four sporozoites are produced (PI. XIII,
figs. 9-11). In many sporocysts it has been impossible to recognise

Fic. 2. Section of a villus of the cat showing Isospora felis in the epithelium and
Isospora bigemina in the deeper tissues. In the epithelium are seen two macrogametocytes,
one cut through the nucleus and one cut to the side of the nuclcus: one microgametocyte
which has given rise to numerous microgametes and a residual body, and one young form
which may be a young macrogametocyte. In the interior of the villus are seen six mature
oocysts of Isospora bigemina. XX 1500.

a residual body, but such a structure is definitely present in some
cases. It appears that it breaks up and disintegrates after the
sporozoites have been formed.

As regards the fate of the fully formed oocysts, there 1s no definite
information to offer, except that they were not detected during the
examination of the faeces made before death. In the sections of the
intestine the epithelium was in many cases absent, so that escape

260

of the sporocysts would be an easy matter if such a change occurred
in life. The heavily infected villi were considerably altered in
appearance. Ihey were swollen, and an excess of cells was present.
It seems probable that such altered villi would break down during
life and liberate the oocysts. [hese would not appear in the faeces
regularly, as in the case of /sospora felis which develops in the
epithelium, but would occur at intervals, whenever a villus broke
down sufficiently to discharge its contents, which would include
oocysts in various stages of development.

The question of a possible relationship between this parasite and
the very much larger /sospora felis which develops only in the
epithelium has been considered. It might be urged that if /sospora
felis developed in the sub-epithelial tissues it might take on the
character of the smaller form. The latter, however, has only been
seen in one animal, while many infected with /sospora felis alone
have been studied. It seems clear, therefore, that the small form is
a distinct species.

The undeveloped oocysts of /sospora bigemina have recently
been detected in the faeces of a dog by Mr. Leslie Sheather, as noted
above.

ISOSPORA RIVOLTA (GRASSI, 1879)

This coccidium has only been studied in the oocyst stage as
found in the faeces of three dogs. Mr. Leslie Sheather has also seen
the oocysts in the faeces of a dog at the Royal Veterinary College.
The oocyst has much the same shape as that of /sospora felis, but
is smaller. The measurements obtained from the three dogs agree
very closely with those given by Grassi (1879, 1882, 1883) for the
form in the cat, and Reichenow (1921) for the one in the dog. ‘The
dimensions given by Wasielewski (1904) for the oocysts seen by
him in the dog are very much the same. as also the smaller series
found by him in the cat. Hall and Wigdor (1918) evidently met
with this parasite in one dog. The reference made by Noller (1921)
to a large and small form in the cat may refer to /sespora felis and
lsospora rivolta.

Four stages of development of the oocyst are shown at Pl. XIII,
figs. 12-15. As seen in English dogs, they vary in length from
20 to 24 microns and in breadth from 15 to 20. Large oocysts

261

like those of the cat are never seen, though, as pointed out above,
when the larger oocysts of Ezmeria canis were present, it was at first
thought that these belonged to /sosfora felis. The difference in size
between the oocysts of /sospora felis and Isospora rivolta was so
constant that there can be little doubt that two species are
represented, as Reichenow (1921) has suggested.

EIMERIA CANIS n. sp.

The oocysts of this coccidium were seen in three dogs, as recorded
by Brown and Stammers (1922). In two of them the infection was
a small one, while in the other it was fairly heavy. The remarkable
feature of the oocyst is its great range in size. In this respect it
resembles Eimeria debliecki of the pig, the oocysts of which were
described by Cauchemez (1921). Another feature which is of interest
in the case of Ezmerza canis is that the sporocysts show the same
proportional variation in dimensions as do the oocysts. It is
evidently incorrect to suppose that in coccidia the sporocysts remain
fairly constant in size in spite of variations in the dimensions
of the oocysts. The oocyst of Ezmeria canis varies in length
from 18 to 45 microns, and.in breadth from 11 to 28 microns.
The general shape of the oocyst will be appreciated from the
figures (Pl. XIII, figs. 16-19, and Pl. XIV, figs. 1-8). . The
cyst wall constantly had a peculiar pink colour, and what seemed
to be the true oocyst wall was enclosed by a somewhat irregular
thick membrane which gradually peeled off during the development
outside the body. When this membrane had separated, the colour
of the cyst was still the same, though much paler. The course of
the development ts illustrated in the drawings. It will be noted that
a definite micropyle could be detected in some oocysts (PI. XIII,
figs. 16 and 18, and Pl. XIV, fig. 1) and that the enclosed cytoplasm
was sometimes attached to it by a strand (Pl. XIII, fig. 16). An
inner membrane indicated by radiating lines could also be detected
in some of the oocysts (Pl. XIII, figs. 18 and 19, and Pl. XIV,
figs. 1 and 2). During the formation of the sporoblasts there was a
striking resemblance to Eimeria stiedae of the rabbit, as described
by Metzner (1903). Pyramidal elevations with clear hyaline apices
were formed. The sporocysts had the characters shown in the

262

drawings (Pl. XIV, figs. 5-8). It will be noted that at the narrower
end there is a definite elevation or knob. In many respects the
oocysts resemble those of the coccidium of the rabbit. Since the
paper by Lucet (1913) appeared, it has been assumed that there are
two coccidia in the rabbit, the one, Eimeria stiedae, with larger
oocysts than the other, Ezmeria perforans, as first clearly stated by
Leuckart (1879). The former, according to Reichenow (1921), who
agrees with this view, occurs in both the liver and intestine. In
some cases the liver alone is infected, in others only the intestine,
while in other animals both are found to harbour the coccidium.
The other form, /zmerza perforans, is apparently limited to the
intestine, though information on this point is not very definite.
There seems, however, no reason to suppose that the coccidium of
the dog represents two species, though in many respects it
corresponds with a mixed infection of two forms in the rabbit. The
great variation in size of the oocysts of Hzmeria canis raises the
question as to whether there are actually two coccidia in the rabbit
or only one.

It does not seem possible to identify the form in the dog with
the common rabbit coccidium, though Bruce (1919) has described a
coccidium of the rabbit in America the oocysts of which resemble
those of Eimeria canis in the presence of the layer of material
covering the wall, in its pinkish orange colour and the marked range
in size. Bruce was inclined to regard this parasite as a new species,
or a variety of the common rabbit coccidium. It certainly resembles
Eimeria canis more than any other recorded coccidium.

It should be mentioned, however, that Guillebeau (1916) has
described a still smaller coccidium, which he says occurs in the liver
cells of dogs. He identified it with Eimeria stiedae, though the
oocysts measured only 12 by 7 microns. As pointed out by
Reichenow (1921), the situation of the parasite in the liver cells 1s a
most unusual one for coccidia. The figures given by Guillebeau do
not assist in arriving at a conclusion as to the nature of the organism.
Chierici (1908), quoted by Martin (1909), recorded a coccidium
which he found in the bile of a cat. The oocysts had a thick,
double-contoured wall, were oval in shape, and measured 26 to
30 microns in iength by 17 to 20 microns in breadth. Development
with the formation of four sporocysts, each with two sporozoites,

263

occurred. It is evidently a coccidium of the genus Eimeria, but
whether it 1s identical with Ezmeria canis cannot be determined.
Virchow (1865, p. 356) records his discovery in the gall-bladder
and bile ducts of one dog of numerous egg-shaped psorosperms with
thick, double-contoured shells. No further description is given, so
that it is not possible to form an opinion as to whether these were
oocysts of coccidia or eggs of a trematode. Another reference to
similar structures is by Rivolta (1878), who gives the name
Cytospermium hepatis canis familiaris to certain oval bodies which
Perroncito had found in the bile ducts of the dog and which he had
called cellule oviforme del fegato del cane. Perroncito (1882, p. 98)
refers to what are evidently these bodies as ‘ Cztospermio del fegato
del cane.’ He gives also the name ‘ Cellule oviforme del fegato
del cane, Perroncito.’ They are described as measuring 48 to
52 microns in length by 21 to 32 microns in breadth. There is a
capsule 2 microns in thickness, and at one-pole an operculum. The
contents divide into two to eight masses. There is little doubt that
these bodies are eggs of a trematode. It appears that the first
reference was made by Perroncito (1876), but this paper has not been

consulted.

ISOSPORA OF MAN

The facts which have been explained above have a direct bearing
on the status of the /sospora which has been recorded from human
beings. It will-be necessary to review the history of the discovery
of the parasite. The first record of the occurrence of such a
coccidium is that of Virchow (1860, p. 527), who mentions a case
which was brought to his notice by Kyjellberg. He found at
post-mortem Psorosperms in the villi, which agreed entirely with
those that he (Virchow) had seen in dogs (‘welche ganz mit
denen tibereinstimmen, die ich beim Hunde gesehen habe’). The
Psorosperms occurred in the interior of the villi, and especially
towards their ends (‘in dem Innern und zwar gegen die Spitze der
Darmzotten’). Of the form seen by him in the dog, he says that
in the interior of the villi he saw numerous Psorosperms of relatively
small size regularly arranged in pairs with a double-contoured
membrane (‘/udess habe ich neulich erwahnt (S. 342), dass ich in
einem Hunde im Innern der Darmzotten sehr hatifig Psorospermien

264

antraf; es waren relativ kleine, regelmdssig su zweien anein-
andergesetzte mit starker, doppeltcontourirter Membran versehene
Korper’). He goes on to state that they must have been like the
forms seen by Finck in the cat. From Virchow’s statements, the
only conclusion justifiable is that he saw in man a small coccidium
like /sospora bigemina of the cat and dog.

The next reference is that by Eimer (1870), but this 1s much less
satisfactory than that of Virchow. Eimer says that he saw
Psorosperms in two men who were examined post-mortem in Berlin.
The intestinal canal was described as being filled and the epithelium
completely infiltrated with Psorosperms. He says they were like
those seen by him in mice and other animals. In both the human
cases the epithelium of the greater part of the intestine is described
as having been devoured by the Psorosperms, as occurs in infected
mice. The contents of the Psorosperms were finely granular.
Eimer furthermore states that he observed all stages of the division
of the contents, but gives no clear account of the process.
From these meagre details it appears impossible to identify the
Psorosperms seen by Eimer. Whether they were coccidia at all is
far from clear. ‘They evidently did not show the same arrangement
in pairs noted by Virchow, for such a striking appearance would
hardly have escaped his notice. The only points in favour of the
view that they were coccidia are the statements that they occurred
in the epithelium, and that they resembled undoubted coccidia of
the mouse and other animals. As coccidia belonging to both the
genera /sospora and Eimeria occur in man, it is fruitless to
speculate as to which genus the form seen by him belongs.

Rivolta (1873) describes certain corpuscles he found in the faeces
of man, but there is no evidence whatever that these were oocysts of
coccidia. Similarly, the bodies seen by Grassi (1879), and which
he regarded as coccidia, were probably cysts of Gzardia._ Rivolta
(1879) proposed the name CyZospermium hominis for the psorosperms
found in man by Eimer. The name is given explicitly to
Eimer’s psorosperms, and Rivolta makes no mention of the bodies
originally described by him in 1873. Thus Rivolta’s name
Cytospermium hominis was given to certain bodies seen by Eimer
which may or may not be coccidia, and even if they were coccidia

are quite unidentifiable.

265

Railliet and Lucet (1890) described the small coccidium of the
villi of dogs. They recognise in these the form named Cytospermium
villorum intestinalium canis by Rivolta (1878). They correctly
followed the development with the production of two sporoblasts,
each of which gave rise to a sporocyst containing four sporozoites.
The oocysts measured 12 to 15 microns by 7 to 9 microns. They
state that they had seen coccidia in the faeces of a woman and her
child who were suffering from chronic diarrhoea. The coccidia
were regularly ovoid, and some of them contained granular proto-
plasm, including a number of refringent globules. Others contained
a large granular mass without globules. The average size was
15 by 10 microns. They recognise, however, that they differed in
certain respects from the forms seen in the dog.

In a later paper, Railliet and Lucet (1891) accept the name
Coccidium bigeminum given by Stiles (1891) to the small coccidium
of the dog. As pointed out above, they recognised three varieties
of this organism, Coccidium bigeminum vars. canis, catt and putorz
in the dog, cat and pole cat, respectively. They say that a fourth
variety probably also exists, namely, Coccidium bigeminum vat.
hominis, the form which was seen by Kjellberg and described by
Virchow (1860). They make no mention of the bodies described by
themselves in 1890. Raiulliet (1895), however, ascribes to the species
Coccidium bigeminum the often quoted parasite discovered by
Kjellberg. As regards the bodies seen by Railliet and Lucet (1900) in
two human cases, Railliet groups them with those described from man
by Grassi and Rivolta as doubtful forms about which it is not possible
to express an opinion. He states, however, that the size of those
recorded by Railliet and Lucet (15 by 10 microns) brings them into
relation with C. bigeminum. Six pages further on in his book,
Railliet again asserts that the parasite discovered by Kjellberg must
without doubt be placed in this species (C. bigeminum), as it was
situated in the interior and towards the tips of the villi, and
resembled the form seen by Virchow in the dog. It is thus evident
that Railliet and Lucet, in employing the name Coccidium bigeminum
var. hominis, were naming not the form seen by themselves, but
Kjellberg’s parasite recorded by Virchow (1860).

From what has been said above, it will be apparent that in only
one of the records, namely that of Virchow, is it possible to make

266

an accurate deduction that a coccidium was being dealt with.
Rivolta’s name Cytospermium hominis refers to Eimer’s parasite
which cannot possibly be identified. If a coccidium at all, it may
have been an /sospora or an Eimeria, but nothing more definite can
be asserted. In the case recorded by Virchow, however, we know
that he was familiar with the small J/sospora of the dog. He
recognised that the latter occurred in the tissues of the villi and not
in the epithelium, and that it occurred in pairs and was like the
parasite of the cat described by Finck. Of the human form, he says
it occurred in the interior of the villi, especially towards their distai
ends, and that it agreed entirely with the one he had seen in the
dog. The only possible conclusion which can be drawn legitimately
from these precise statements is that Virchow actually meant what
he said and was observing in man a small /sospora like /sospora
bigemina of dogs and cats. With the very doubtful exception of
the bodies seen by Railliet and Lucet (1890) this small coccidium
has not since been discovered. At first sight this may seem
surprising, but there appears to be a possible explanation. When
Finck made his observations on the cat he was concerned mostly
with the changes undergone by the intestinal epithelium during
digestion rather than with the faeces. He was actually examining
the intestinal wall itself, and not the dejecta of his animals.
Similarly, Virchow and Rivolta, who saw the small /sospora of
dogs, were concerned mostly with the wall of the intestine, and the
same appears to be true of Railliet and Lucet, and Stiles. As
pointed out above, the presence of /sospora bigemina in the cat was
only detected by the writer when scrapings were made from the
intestinal wall. In these scrapings the thick-walled sporocysts, often
arranged in pairs enclosed by a common membrane, were very
striking objects, whereas the incompletely formed oocysts of the
large Isospora felis which were also present were not nearly so
easily seen, and might readily have been mistaken for enlarged tissue
cells. If examination in this case had been limited to the faeces
the small forms would have been missed entirely, and only the
oocysts of the large form seen.

Grassi, however, was concerned largely with the examination of
the intestinal contents and faeces, with the result that he discovered
the oocysts of the intermediate sized /sospora rivolta in the cat.

267

When he examined the intestinal epithelium he noted that they were
present in the epithelial cells, but there was no indication of a paired
arrangement as in the case of the small /sospora bigemina seen by
Finck and others. Since Grassi’s time, Wasielewski and other
observers, who have likewise studied the faeces, have noted in cats
and dogs both /sosfora rivolta and Isospora felis, but never the
small /sospora bigemina. ‘The developmental stages of the larger
forms have been seen only in the epithelial cells, and never in the
paired condition in the tissues of the villi. It is not improbable that
the tissue-invading small form has been frequently missed owing to
failure on the part of investigators to examine scrapings from the
intestinal wall itself. Virchow discovered the small form in man
because he adopted this method, and it 1s probable that it would
have been re-discovered in recent years had this practice been
continued and if examinations had not been limited to the faeces
alone.

During the extensive examination of faeces of men necessitated
by the exigencies of the war, the oocysts of an /sospora were
discovered on many occasions. ‘They were first seen by Woodcock
(1915) and then by the writer (1915), who demonstrated their
development and proved that they actually belonged to the genus
Isospora, as had been suggested by Woodcock. In a recent paper,
Connal (1922) has shown that over one hundred and fifty cases of
infection with this parasite are on record. The oocysts measure
from 25 to 30 microns in length by about 12 to 15 in breadth. They
thus correspond in size with those of /sospora rivolta of cats and
dogs. They differ, however, in shape, so that they cannot be
identified with the parasite of dogs and cats. From what has been
said above, it is evidently impossible to identify this human
Isospora with the small form (/sospora bigemina) of cats and dogs
or with the small form (/sospora hominis) seen by Virchow in man.
The fact that the oocysts appear in the stool in the undeveloped
condition is strongly suggestive of a development in the epithelium
like Jsospora rivolta and Isospora felis of cats and dogs.

Dobell (1919), in his careful review of the coccidia of man, based
his arguments on the assumption that only one /sospora occurred in
cats and dogs, and under the name /sospora bigemina he included
the small, intermediate and large-sized forms of these animals.

268

Hence in his discussion of the name which should be applied to the
Isospora of man, with every justification he included under the
name /sospora hominis the small form described by Virchow and
the much larger form discovered during the war. When it is
realised that the small form in cats and dogs which develops in the
tissues of the villi is distinct from the larger forms which develop
in the epithelium, this position as regards the human parasites at
once becomes untenable. The small /sospora of man described by
Virchow was named /sospora bigemina var. hominis by Railliet and
Lucet (1891), a name which becomes /sospora hominis Railliet and
Lucet, 1891. As we have seen, the name Cytospermium hominis
of Rivolta was given to unidentifiable structures seen by Eimer
(1870). Dobell (1919) recognises this latter fact, but adopts the
position that it is better to assume that Eimer was actually dealing
with the form described by Virchow, and strongly urges that this
view be accepted. But this statement was made on the assumption
that the small forms in the dog and cat were identical with the
larger ones, an attitude which is maintained by Dobell and O’Connor
(1921), who employ the name /sospora rivoltae. It seems unwise
to make this assumption, as there are absolutely no data to indicate
the nature of the structures seen by Eimer. It is more logical to
adopt the name /sospora hominis Railliet and Lucet, 1901, for the
small /sospora of man, and to regard Rivolta’s name Cytospermium
hominis as a nomen nudum.

As regards the large /sospora of man, no special name has been
given to it, though Savage and Young (1917) employed the term
Coccidium isospora for this parasite. As pointed out by Dobell
(1919), this is evidently a misprint or /apsus calami. The intention
of the writers was not to introduce a new name, but to refer to a
coccidium of the genus /sosfova in contradistinction to one of the
genus Eimeria, as coccidia belonging to both these genera had been
recorded from man during the examinations for intestinal protozoa
made during the war. If, however, it is claimed that the name
was correctly presented, then, Coccidium being a synonym of
Eimeria, Savage and Young’s name becomes /imeria isospora, and
one would have to conclude that they were recording an Eimeria of
man. There is actually no evidence in the paper that this was not the
case, however improbable such a conclusion may be. Their name 1s,
strictly speaking, a momen nudum.

269

An appropriate name for the /sosfora of man which figured so
largely during investigations on the intestinal parasites of man
conducted during the war would be /sospora belli. It may at first
sight appear to cause confusion to introduce a new name for a
parasite which is now generally known as /sospora hominis, but
Virchow (1860) so definitely referred to a small /sosfova of man,
which was named Coccidium bigeminum var. hominis by Railliet and
Lucet (1891), that to submerge this form by applying the name to a
much larger and evidently distinct species which is perhaps more
easily detected, is not only contrary to scientific procedure, but is
unfair to its discoverer and misleading to future investigators. It
seems highly probable that if the method of examination of the
small intestine at post-mortem by scrapings from the wall be adopted
as a regular procedure the small /sospora hominis, first seen by
Kjellberg, will be re-discovered.

CONCLUSIONS

1. here occur in cats and dogs three species of coccidia of the
genus /sospora, namely, /sospora felis n.sp., Lsospora rivolta
(Grassi, 1879) and J/sospora bigemina (Stiles, 1891). The last
named is a small parasite of the deeper tissues of the villi of the small
intestine, and development of the oocyst may be completed in the
vertebrate host, while the two former are larger and are parasitic in
the epithelium covering the villi, the development of the oocysts not
taking place till they have left the body.

2. It is possible, as maintained by Railliet and Lucet, that there
are different varieties of /sospora bigemina, namely, 1. bigemina
vars. canis, cati and putori from the dog, cat and pole cat,
respectively, but there is at present insufficient evidence to justify
the conclusion that they are distinct.

3. The large parasite of the ‘ swift fox,’ described by Weidman
as a possible variety of /sospora bigemina, does not belong to this
species, but is more nearly related to /sospora felis. If it is a new
species, its name will be /sospora canivecolis.

4. The complete development of /sosfora felis in the epithelium
is described. A characteristic feature of the intracellular stages is
the gregariniform character of the parasite. Schizonts produce, as

270

a rule, eight merozoites, but sometimes a larger number. The nuclei
of the microgametes are the result of repeated division of the original
single nucleus of the young microgametocyte. The karyosome
appears to be present in all stages of growth of the parasite. The
oocyst wall is not completely formed till the parasite has left the
cell, and no change in its contents occurs till the oocyst has left the
body.

5. The complete development, including schizogony and
sporogony, of /sospora bigemina takes place in large cells in the
internal tissues of the villi, and here the oocyst is formed and
completes its development. Its wall is comparatively thin, while
that of the sporocyst 1s relatively thick.

6. The development of the oocyst of /sospora rivolta was
studied, and this takes place only after it has left the body, as in
the case of /sospora felis.

7. The parasite described from the interior of the villi of man
by Virchow is a small /sospora like Isospora bigemina. It bears
the name /sospora hominis (Railliet and Lucet, 1891).

8. For the larger form discovered in the faeces of man during
the war, and regarded by Dobell as identical with the small form
described by Virchow, the name /sospora belli n. sp., 1s proposed.

g. A coccidium of the genus Azmeria is described from the
faeces of dogs. This form is remarkable in that the oocysts vary
considerably in size. The name Eimeria canis n.sp., 1s proposed

for this parasite.

The writer is much indebted to various people for assistance
rendered ; especially to Dr. G. Lavier, of the Faculté de Médecine, —
Paris, who very kindly obtained information regarding certain
papers which could not be found in London.

271

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275

ADDENDUM

Since the foregoing account of the coccidia of cats and dogs
was written, a paper has come to hand by Zapfe dealing with the
Isospora of dogs in Germany. The form studied appears to be the
one of intermediate size mentioned by Reichenow (1921), and which
has been identified as /sospora rivolta. It was assumed above that the
development of /sospora rivolta would be found to take place in the
intestinal epithelium, and this has been demonstrated by Zapfe.
The various stages are very similar to those of /sospora felis, but
they are correspondingly smaller, as was to be expected from the
smaller size of the oocyst. During schizogony from eight to
twenty-four merozoites are produced. The infection is as a rule
limited to the distal ends of the villi, as in the case of /sospora felis.
Zapfe regards the parasite as /sospora bigemina, and discusses the
statements that have been made as to the occurrence of oocysts in
the interior of the villi. He inclines to the view that the oocysts
are not actually in this situation, but only appear to be there on
account of irregularities in the epithelium. It is evident he has not
encountered the small form which unquestionably develops in the
interior of the villi.

Reference 1s made to a paper by Pospiech (1919), which the
writer has not seen. This author examined the faeces of a large
number of dogs, and came to the conclusion that there were actually

| four types of /sospora in cats and dogs. Three of these correspond

with the three forms described above. A fourth type, which occurs
in both cats and dogs, has an occyst which varies in size between
that of /sospora bigemina and Isospora rivolta. The dimensions
are given as 17 to 18 microns by 14 microns. The size of the
sporocyst is 11 by 7°5 microns. The writer has not seen this form
in England, and can express no opinion as to whether it is a distinct
species. Zapfe also mentions a paper by Bornhauser (1912), who
described a coccidium of the liver of dogs. No6ller is quoted as

_ having expressed the opinion that the structures described were

276

probably not parasites at all. Reichenow (1921) has come to the

same conclusion.
A paper by Otten €1923) refers to the separation of oocysts from

the faeces of dogs by a saline concentration method. |

REFERENCES

Bornuauser, H. (1912). Leberkokziosis beim Hund. Dissertation, Bern.

Orten, L. (1923). Die Kochsalzmethode bei der untersuchung der Haustierkokzidien. Inaug.
Diss. Berlin.

Posprecu, W. (1919). Untersuchungen tiber den mikroskopischen Nachweis von Darmparasiten
im Kot von Fleishfressern, mit besonderer Berticksichtigung der Kokzidien ©
Veterinarmedizen. Inaug. Diss. Munchen.

Zavrr, H. (1923). Zur Kenntnis der Kokzidiose des Hundes. Inaug. Diss. Berlin.

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EXPLANATION OF PLATE IX
Isospora felis. (XX 2000.)
Figs. 1-18. Schizogony.

Smallest form in vacuole in epithelial cell showing attachment to surface of
vacuole.

Slightly larger form with similar attachment.
Stage just prior to commencement of nuclear division.
Commencing nuclear division. The karyosome in division.

Intranuclear spindle showing equatorial plate of chromosomes and daughter
karyosomes at ends of spindle.

Similar stage showing daughter plates of chromosomes.

Stage with two nuclei.

Similar stage.

Second nuclear division.

Stage with four nuclei.

Third nuclear division.

Stage with eight nuclei. The karyosome is still present though reduced
in size.

Formation of merozoites from the central cytoplasmic body. Only six of
the eight merozoites are shown.

Eight merozoites and residual body in vacuole in cell.

Eight merozoites of larger size in vacuole.

Three merozoites in a vacuole. ‘This is either division into a small number
of merozoites or the result of multiple infection.

Stage with sixteen merozoites, only fourteen of which appeared in the section.

Stage with sixteen larger merozoites.

Figs. 19-28. Growth of microgametocyte.

Young microgametocyte ?

First nuclear division.

Similar form.

Stage with two nuclei.

Stage with four nuclei.

Stage with eight nuclei.

One section of stage with sixteen nuclei.

One section of stage with about thirty-two nuclei.

One section of stage with larger number of nuclei, many of which are dividing.
The chromosomes can be detected.

One section of stage with still larger number of nuclei.

PLATE 1X

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EXPLANATION OF SPLAT E SX

Figs. 1-7. Growth of Microgametocyte (contd.)

One section of stage in which the chromatin has become more distinct, the
cytoplasm clearer and the karyosome smaller. Definite mitotic division
of the nuclei is taking place.

One section of stage in which the chromatin is still more marked.

One section of stage in which the chromatin is much coarser. Some nuclei
are showing what is probably the last nuclear division.

One section of stage in which the final nuclear division has taken place.
Each nucleus includes several coarse chromatin masses. In some an
isolated granule can be detected. ‘This may be the karyosome.

One section of stage in which the chromatin granules are becoming aggregated
into a single mass.

One section of stage in which the chromatin of the nuclei has become com-
pletely condensed into a single mass and has formed finger-like outgrowths.

One section of stage in which the chromatin of the nuclei has assumed a
falciform shape.

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' a Bxoplg!.cowy oe heanebar et Sout af T brerighicrig ‘

wee Srl, teadeys bartersin {ehrnarg alr igeahee » ne to ;
-, Y '
a 7 » + i
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at FOS 1 te , ,
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= =
ne
: a is
_

282

EXPLANATION OF PLATE XI

Fig. 1. One section of stage in which microgamete formation is completed and

IO.

Miho

several residual bodies are present.

Figs. 2-11. Growth of macrogametocyte.
Very young macrogametocyte ?

Slightly later stage showing attachment to the surface of the vacuole against
the nucleus of the host cell.

Later stage showing attachment to the nucleus of the host cell, which has
been drawn into the vacuole.

Still later stage showing the appearance of a terminal sucker into which a
pedicle of the cell cytoplasm has been drawn.

Still later stage attached to nucleus.
Section of larger form with nucleus of host cell within the vacuole.

Section of larger form showing doubled-up condition. The granules of
deeply staining material are appearing round the nucleus.

Section of later stage. The granules round the nucleus are more markéd
while deeply staining masses appear in the cytoplasm.

Section of larger form. Globules of a refractile substance are appearing in
the cytoplasm.

Section of a stage in which the globules of refractile substance are more
pronounced. The surface is indented in two places by an accumulation
of an eosinophile granular material against the wall of the vacuole.

Bina XL

Annals Trop. Med. & Parasitol., Vol. XVII

C. Tinling & Co., Ltd., Imp.

THE LIB ARY
OF THE ;
Ment FRSITY OF WLteais

Fig.

284

EXPLANATION OF PLATE XII

1. Fully developed stage with clear cytoplasm filled with globules of refractile
substance. ‘The oocyst wall is just commencing to form.

Figs. 2-11. Sporogony.

A srviall area of cytoplasm is figured round each nucleus in Figs. 2-7.
Nucleus of the zygote.
Two nuclei in zygote after first nuclear division.
First nuclear division in a sporoblast.
Two nuclei in a sporoblast.
Second nuclear division in a sporoblast.
Four nuclei in a sporoblast.
Sporozoite showing granule at centre of karyosome.
Sporozoite with karyosome more deeply stained.
Sporozoite showing vacuole in cytoplasm left by solution of refractile body.

Stained sporocyst showing four sporozoites and large residual body.

Figs. 12-15. Isospora felis—oocysts as seen in living condition. (xX I500).
Condition in which oocyst leaves the body.

Oocyst in which the zygote has become spherical and the nucleus divided.
Two sporoblasts in which first nuclear division is taking place.

Mature oocyst showing two sporocysts, each with four sporozoites and a

residual body.

PLATE XI

Annals Trop. Med. & Parasitol., Vol. XVII

Imp

C. Tinling & Co., Litd.,

THE LIBRARY .
OFTHE
UNIVERSITY AE 11 UNOIS

mx sae FO “OITA MAITX: a
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bal a a .
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me -

wy tts oe si sfogvs SAT «6 doeIni gaitsvo9 ta1W0 Aft i200 1 =

eoibod lnbimetyq ex etexid

286

EXPLANATION OF PLATE XIII
Figs. 1-11. Isospora bigemina. (X 2000).

1. Two young schizonts in mononuclear eell.

2. Multinucleated schizont.

3. Commencing segmentation of schizont.

4. One section of stage with about sixteen merozoites.

5. Microgametes and residual body.

6. Partly developed macrogametocyte.
7. Fully grown macrogametocyte.
8. Odocyst with enclosed zygote.

9g. Odocyst with two sporoblasts.

10. Fully developed oocyst with two sporocysts, each with four sporozoites and
a residual body.

11. Similar stage with no residual body visible in the sporocysts.

Figs. 12-15. Isospora rivolta—oocysts as seen’in living condition. (X 1500).
12. Condition in which oocyst leaves the body.
13. Oocyst in which zygote has become spherical.

14. Oocyst with two sporoblasts in one of which the nucleus is dividing, while
in the other the first nuclear division is complete.

15. Mature oocyst containing fully developed sporocysts.

Figs. 16-19. Eimeria canis—oocysts as seen in the living condition. (X 1500),

16. Large oocyst with spherically contracted zygote attached to micropyle by
pedicle.

17. Very much smaller oocyst of similar type.
18. Large oocyst with the outer covering breaking away.

19. Oocyst with outer covering intact. The zygote is budding off from sporo-
blasts as pyramidal bodies.

PEA eee Lb

Annals Trop. Med. & Parasitol., Vol. XVII

C. Tinling & Co., Lid., Imp.

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at ‘ Js tt OCA ie i PP saJdRT Jit sth se} i i . }

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288

EXPLANATION Ob SPI ay

Figs. 1-8. Eimeria canis—oocysts as seen in the living condition (continued).

(x 1500)
Oocyst with outer covering intact. Four sporoblasts and a residual body
are present.
Similar form with four sporoblasts and no residual body.

Oocyst with four elongated sporoblasts and a residual body. The outer
covering of the oocyst has disappeared except at two small areas.

Oocyst in similar stage of development with outer covering intact.

Oocyst with outer covering intact and four undeveloped sporocysts and a
residual body.

Completely developed oocyst with residual body and four sporocysts, each
of which has a terminal knob and includes two sporozoites and a residual

body.
Completely developed oocyst of much smaller size.

Similar but slightly smaller oocyst.

Annals Trop. Med. & Parasitol., Vol. XVII PLATE XIV

C. Tinling & Co., Ltd., Imp.

289

A FURTHER NOTE ON THE OCCUR-

RENCE OF ANCYLOSTOMES RESEMBLING

NECATOR AMERICANUS AMONGST
DOMESTIC PIGS IN AMAZONAS

BY
R. M. GORDON

(Received for publication, April 25, 1923.)

In a previous note (1922) the author gave a brief description of Necators
obtained from domestic pigs in Amazonas and reached the conclusion
that, although of smaller size, the worm was indistinguishable from the
human parasite Necator americanus. About the same time Ackert
and Payne (1922) described a hookworm from the gut of the domestic
pig in Trinidad. They stated that although this worm resembled Necator
americanus, it exhibited certain differences which in their opinion were
of specific value and accordingly they gave it the name Necator suillus.
They elaborated their arguments in a later paper (1923). The points on
which they base their conclusion that the pig worms differ from the
human are the following :—

(1) ‘ The new species is somewhat smaller.’

(2) ‘The buccal capsule much smaller proportionately.’

(3) ‘As a rule the dorsal turn in the neck is not so pronounced as
in Necator americanus.’

(4) ‘In N. swillus the lateral lancets are broadly wedge-shaped in
profile, while those of the large Necator are cusp-shaped. The ventral
lancet is slender in side view pointing towards the base of the dorsal
tooth in N. swillus while this lancet in the larger species is broader and
points approximately towards the tip of the dorsal tooth.’

(5) ‘The dorsal rays in the new species are shorter, while their

290

terminal branches are actually longer than these structures in the larger
N. americanus.’

(6) ‘ Incomparing specimens of these two species, cleared in glycerine,
one is struck by the large so-called body cavity in the females of
N. americanus, as contrasted with a much smaller one in WN. swallus.’

(7) ‘Concerning the males of these two species, the most striking
differences are the proportions and shape of the bursa when closed.
In N. americanus this organ is about as long as wide, and is distinctly
funnel-shaped, the distal edges being flared like the bell of a trumpet,
while the bursa of N. swillus is distinctly longer than wide and is more
cup-shaped.’

(8) ‘A conspicuous difference between these two species seen under
higher magnification is the form of the head papillae. In N. sullus
each lateral papilla unites with the corresponding dorso-lateral one,
enclosing a conspicuous cam-shaped depression, a condition not true of
N. americanus. Further it may be noted that in N. americanus the distal
ends of the dorsal, lateral and ventral papillae are more or less beaded or
constricted, while in N. swillus no such structures occur at the ends of
the papillae.’

(9) ‘Another rather constant difference is the shape of the externo
dorsal ray. In N. suillus this ray, which is of nearly equal width through-
out its length, makes a sharp lateral turn near its distal end; while in
N. americanus the width of this ray is variable and the turn at the tip is
less pronounced.’

(10) ‘Finally the spicules show constant differences. The average
length of the spicules of N. americanus is double that of the spicules of
N. suillus. In the latter species both shafts terminate distally in the
membranelle as recurved hooks, while in N. americanus only one shaft
ends as a recurved hook, the other terminating in a nearly straight line.’

In view of the work of Ackert and Payne the writer has re-examined
the pig ancylostomes from Amazonas and compared them with ancylos-
tomes obtained from the human host in Amazonas and Jamaica, with
special attention to the points mentioned above.

(1) Length of the two worms. Table I shows that, whereas the average
size of the pig ancylostome is distinctly smaller, yet its maximum length
is equal to the minimum length of the human parasite from Jamaica,
and greater than that of the human parasite from Amazonas. The length
cannot therefore in itself be used as a distinguishing character.

—

291

Taste I,

Showing the lengths of Necators obtained from pig and human hosts.

te Amazonas

human host, Jamaica

> human host, Amazonas ...

nan host,
9 a eee
a

nan host,
ldinas ..,

—

}, Amazonas...

Males Females
Maximum Minimum | Average Maximum | Minimum | Average
Number | length in | length in | length in | Number | length in | length in | length in
measured | millimetres millimetres| millimetres) measured | millimetres| millimetres) millimetres
28 6°5 4°5 st 64 8-2 5°5 6°5
28 g°0 6°5 7°8 64 13°0 8°5 10°9
28 ee ae: 6°8 64 11'S a g'I
|
(2) Size of buccal capsule. It appears from Table IT that the buccal
: capsule is proportionately greater in the pig than in the human
ancylostome ; this is the reverse of Ackert and Payne’s findings. Much
reliance cannot, however, be placed on small differences in these measure-
ments, as well-marked variations in shape from the normal oval of the
Taste IT.
Showing the measurements of the buccal capsules in Necators obtained from pig and human hosts.
Males Females
Maximum Minimum | Average Maximum Minimum Average
Number — ——— | Number —— ——— <<) Semone
measured |Dorso- Dorso- Dorso- measured |Dorso- Dorso- Dorso-
ventral|| ateral|ventrall, ateral|ventral Lateral ventrall { ateral|ventral| Lateral|ventral|Lateral
diam. | diam. | diam. | diam.|} diam. | diam. diam. | diam. | diam. | diam. | diam. | diam.
be td LU Nd ed ad ed od be sd Lb it
14 66) 59 “ar592| 55.7). 09. ies 31 Sih 7e sy 06" 159°) 63" | 157
15 74 57.) Peele 44 Do RSS 15 Bar, OS) FEZ. | 54h oF | BOI
IS 64 61 51 44 61 46 15 85 68 64. 47 74 60

mouth capsule were frequently encountered ; another difficulty is that
any variations in the angle from which the head is viewed will give rise

to different results in the measurement of the mouth capsule.

(3) Dorsal curvature of anterior part of body. No such constant

differences as those described by Ackert and Payne were observed in the

anterior curvature.

292

(4) Ventral lancets, lateral lancets and dorsal tooth. An examination
was made of a large number of worms from both pig and man, but no
constant differences in the lancets or dorsal tooth were found ; the dorsal
tooth and the ventral and lateral lancets of both worms showed great
variation in size, shape, and angle of projection, as is illustrated in fig. 1,

ehes g
cra B . oy
eet

Sen

Fic. 1. The three projections in each drawing from above downwards are V./.—Ventral
lancet; L.J.=Lateral lancet; D.t.=Dorsal tooth. A, B, C, D, E and F=Necator americanus from
human host, Amazonas; G, H, I, ¥, K and L=Necators from domestic pig, Amazonas.

which represents camera lucida outlines drawn from unselected material.
With regard to the angle of projection of the ventral lancets it is of interest
to note that Ackert and Payne in their last paper depict these lancets
as projecting at almost precisely the same angle in both N. americanus
and N. suillus.

293

(5) Length of the dorsal rays.

The results of measuring the dorsal

rays in eighteen worms from pigs and in twenty-four from human material

are shown in Tables III and IV.

ABT rel LL

Showing measurements of dorsal rays and their branches in Necators obtained from pig and human hosts.

ig, Amazonas 95°0 650 80°0 34°0 | 180 26°4 160
suman host,

veces 130° 97'0 III'S 36°4 26°0 29°9 19'0
suman host,

zonas 120°0 78°5 110°5 32°5 19°5 25°9 15"0

Taste IV.

10"4

Length in microns
of outer branch of dorsal ray

13°9

11°8

Ratio of length
of dorsal ray
to total length of worm

Ratio of length
of inner branch of dorsal ray
to total length of worm

Maximum Minimum Average | Maximum|/ Minimum | Average
ig, Amazonas rhe: I: 92 1:71 1: 183 a Ree roeeie
‘uman host,
ica Ti: 59 1: 80 1:69 I: 206 I: 307 1er257
uman host,

ooh E 1: 89 #365 I: 224 1-8-357 let-277

Ratio of length
of outer branch of dorsal ray
to total length of worm

Maximum

F-24134

I: 343

1: 394

Minimum

> 528

27726

1023

From Table IV it follows that the average of the ratios of the lengths
of the branches of the dorsal ray to that of the worm is slightly greater

in the case of the pig Necators than in the human, but the maximum

and minimum values overlap to such an extent as to make this point of

no specific value.

From Table III it can be seen that Ackert and Payne’s

statement that the actual lengths of the branches of the dorsal ray are

greater in N. sutllus, does not hold good for the Amazonas material.

(6)

Size of the body cavity. Eight worms from the pig contrasted

I: 402

T4538

I: 610

. | . .
Maximum Minimum] Average

owing ratio of length of dorsal ray and its branches to total length of the worm in Necators obtained from pig and human hosts.

Average

2
with eight N. americanus from the human host in Brazil, showed no

proportionate difference in the size of the body cavity.
(7) The
dimensions of the closed bursa from its ventral aspect, of six Necators

The shape and proportions of the bursae in the two worms.

from the pig were 0:26 mm. long by 0:24 mm. broad, while those of six
Necators from the human host in Brazil were 0-38 mm. long by 0.41
mm. broad. It is difficult to say when a bursa is completely closed
and various stages between cup and funnel shape were observed both
amongst Necators from the human and from pig hosts, but these
variations in shape appear to depend entirely on the degree of
approximation of the two halves of the bursa.

(8) The head papillae.

papillae as described by Ackert and Payne was clearly visible in the

The union of the lateral and dorso-lateral

majority of the pig specimens, in others, however, no such union could be
traced ; it was, moreover, also present in many of the Necators of man,
though possibly not as often as in those of the pig. The beading on the
papillae was also found to be a variable factor and was seen at times
in both worms.

(10) Length and shape of the spicules.
spicules of twelve worms from pigs in Amazonas and of those obtained from

The results of measuring the

the human host in the same locality are recorded in Table V.

TABLE V.

Measurements of the spicules of Necators obtained from pig and human hosts in Amazonas.

Length of worm in millimetres

Length of spicules in millimetres
| 1

Ratio of length of spicul

to total length of worn
|

—

Avi

From pig, Amazonas ...

From human host,

Amazonas

It appears from Table V that the average length of the spicules in
the worms from man are nearly double the length of those in the worms
from the pig. The average of the ratios of the length of the spicules to
that of the body are, however, respectively 1:8 and 1:11, and it must
moreover be noted that the maximum ratio in the case of the pig (1: 8:4)

is greater than the minimum ratio in the case of man (I : 10-6),

295

and consequently this can hardly be regarded as a reliable point of
distinction. Ackert and Payne’s statement that both spicules in the
pig Necator terminate in the membranelle as recurved hooks in contrast
to N. americanus in which only one spicule is hooked, was not found to
be constantly true of the worms from Amazonas, one hooked and one
nearly straight spicule being very common amongst the pig Necators,

5-7

Fic. 2. 4, B, C and D=Spicules of Necators from domestic pig, Amazonas.

E, F, Gand H = Spicules of Necator americanus from human host, Amazonas.

while in the case of the Necators from man both spicules frequently
showed well-marked hooks. These points are illustrated in fig. 2.

Size of ova. Ackert and Payne give the average size of the ova of
the pig Necators from Trinidad as 63 by 37, the ova of the pig Necators
from Amazonas were found to measure on an average 64 by 39/.

296
ATTEMPTS TO INFECT PIGS WITH NECATOR AMERICANUS

Ackert and Payne (1923) performed several experiments to ascertain
whether N. americanus from the human host can mature in pigs ; they

also carried out experiments in pigs with cultures of N. swillus. For the |
sake of brevity, the results of Ackert and Payne’s experiments are con-
densed in Table VI. |

Taste VI.

Summarising the results of Ackert and Payne’s attempts to infect domestic pigs with larvae of N. americanus obtaine:

from human sources.

L
No. of | No: of Date Stage How given Date Result Remarks |
pig larvae given of larvae killed of autopsy
I 2841 16.6.21 Not stated 10.8.21 | 3 adult Faeces free from Nematc
2700 20.6.21 ‘Infective ’ N. suillus ova or larvae at time’
3000 27.6.21 f experiment |
2 2841 16.6.21 | ‘Infective’ Not stated 10.8.21 | 8 adult Faeces free from Nemat¢:
N. suillus ova or larvae at time!
experiment
Pedr Ad oe EF ee es a
3 1850 27.9.21 By mouth on 7 adult In spite of oil of Cher.
12000 29.9.21 ‘ Infective ’ bread q:i222 N. suillus podium treatment, f}
4000 4.10.21 3, 4, and 5 alla
Ancylostome ova at ti!
of experiment :
4 7500 29.9.21 | Not stated Placed on 3-12.21 | 6 mature

ggoo 1.10.21 | Sheathed shaven skin N. suillus | |
5 Nil — — -~ 3.12.21 | 3 adult Used as a_ control
N. suillus pigs 3 and 4 |
6 5000 ‘Infective’ Not stated 73 days No hook- Owing to the difficulty!
after worms found) obtaining pigs free fri
infection ancylostomes in Trinid,
this experiment was ci’
ducted in Manhatt,

Kansas

Ackert and Payne claim to have shown from these experiments that
it is not possible to infect pigs with Necator americanus, either by the
mouth or by the skin. This claim appears to the writer to rest entirely
on their ability to distinguish between Necator americanus and Necator
suillus, for in every experiment, except in the case of Pig 6, where the |
method of administering the larvae is not stated, N. suwillus was found
at the autopsy. It is true that the number of worms found is small (3 to 8),

Results

297

but on consulting Table VII, it will be seen that equally large doses
(6,900 and 23,000) of infective larvae of N. swillus resulted in an almost
equally small production (10 to 32) of adult worms. It appears certain

Tasre VII.

of Ackert and Payne’s experiments at infection of domestic pigs with larvae of N. swillus obtained from
naturally infected pigs.

No. of Date Stage How given Date Result Remarks
larvae given of larvae killed of autopsy
|
2500 29.9.21 | Infective : By mouth __—Died 32 Pigs 1 and 2 both showed
4400 1.10.21 5-11.21 N. suillus ancylostome ova in their
| ——|_—————_|_ faeces previous to the
11000 9.10.21 Other: Placedon = 3.12.21_| 10 adult experiment for which
12000 | II.10.21 shaven skin — N. suillus they received anti hel-
| minthic treatment the
results of which are not
| recorded
|

therefore that it is extremely difficult to infect the domestic pig with
Necators whether the infective larvae used are obtained from other
pigs or from the human host.

The present writer while in Amazonas undertook no experiments to
infect pigs, owing to the difficulty of obtaining pigs free from Necators
and to his inability to distinguish between the pig and the human Necators.
One experiment has since been carried out in Liverpool. A pig six weeks
old was obtained and kept under observation for seven days ;
during this period its faeces were examined daily by the
saturated salt method (Willis, tIg21), but with negative results.
On 15.12.22 approximately 400 sheathed larvae obtained from
a culture of human faeces were mixed with 2 c.c. of Normal saline
and injected subcutaneously in the back of the pig, a similar dose was
again given on 29.12.22. The patient from whom these cultures were
made was later treated with Carbon tetrachloride and all the 96 worms
obtained proved to be N. americanus, it therefore appears reasonably
certain that the larvae administered to the pig were those of Necator
americanus. Tyrichuris ova made their appearance in the faeces shortly
after the first inoculation and persisted throughout the experiment ;
no other ova were seen till 19.1.23, when ancylostome-like ova first
appeared in the faeces. These ova which were very regular in contour and
size, measured 68 by 37m, and were quite indistinguishable from those

298

of the human ancylostomes ; they were always very scanty in numbers,
never more than three being found in a single cover slip preparation
made from about two grammes of faeces treated by the saturated salt
method. Ancylostome ova were not always found at these examinations,
the faeces being sometimes negative for two, or even three days. The last
occasion on which ova were found present was 5.2.23, examinations on
the subsequent four days being negative. The pig was killed 9.2.23, and
in spite of a very careful search of the oesophagus, stomach, intestines,
trachea, bronchi, etc., the only helminths found were numerous Trichuris
in the caecum. During the last week of the experiment the whole
bulk of the faeces was daily examined for any Ancylostomes that might
be passed per rectum, this search proved negative, but it is extremely
difficult to detect an odd Ancylostome in a large mass of faeces and it is
therefore uncertain whether the pig got rid of the worms responsible for
the ancylostome-like ova during the four days prior to the autopsy, or
whether they were missed at the post-mortem; as the search of the
organs was undertaken with great care the former conjecture appears

the more likely.

CONCLUSION

No constant differences were found between the Necators of Amazonas
pigs and those of man from Amazonas and Jamaica such as would justify
the formation of a new species for the pig worm.

REFERENCES

AckeERT, J. E. and Payne, F. K. (1922). Investigations on the control of hookworm disease.
V. The domestic pig and hookworm dissemination. Am. four. Hyg., Vol. Il, pp. 39-50.

AcxerT, J. E. and Payne, F. K. (1923). Investigations on the control of hookworm disease.
XII. Studies on the occurrence, distribution and morphology of Necator suillus including
descriptions of the other species of Necator. Am. Four. Hyg., Vol. II, No. 1, pp. 1-25.

Gorpon, R. M. (1922). The occurrence of Ancylostomes resembling Necator americanus amongst
domestic pigs in Amazonas. Ann. Trop. Med. and Parasit., Vol. XVI, No. 3, p. 295.

Wituis, H. Hastines (1921). A simple levitation method for the detection of Hookworm ova.
Med. Four. of Australia, Vol. 11, No. 18, p. 375.

i pe

A CASE OF ACUTE ASCENDING
PARALYSIS IN A CHIMPANZEE

Sp we DIDS
AND

E. J. CLARK

(Sir Alfred Lewis Jones Research Laboratory, Freetown)
(Received for publication 15 November, 1922)

A male chimpanzee (Anthropopithecus troglodytes), judged to
be about four years of age, was captured by natives on the
3rd of September, 1922, near Blama, Sierra Leone.

The animal appeared healthy and showed no evidence of injury
to the spine. On the evening of the 5th of September the animal
was chloroformed in order to be caged, and was then sent down to
Freetown. Very little chloroform was used and the animal quickly
recovered from the anaesthetic.

On arrival in Freetown, on the 7th of September, two abrasions
in the loins, due to the chafing of a rope, were observed; these
quickly improved on the application of iodine.

On the 15th of September it was noticed that the animal refused
to leave its cage and did not take food; on removal from the cage
it was found that the lower limbs were completely paralysed and
were colder than the rest of the body. The animal eagerly drank
large quantities of milk and water, but refused solid food.

On the evening of the 17th of September the trunk and the upper
limbs were completely paralysed; the face was cyanosed and the
animal suffered from dyspnoea. Mulk feeds were vomited. The
muscles of the neck were not affected and the animal could move
its head freely from side to side.

On the 18th of September o'09 gms. of Novarsenobillon were
administered intramuscularly into the thigh; there was no loss of

300

sensation and the animal moved its head vigorously from side to
side and attempted to seize the hand of an assistant with its teeth.
The animal’s condition appeared to improve rapidly after the
administration of Novarsenobillon; the cyanosis disappeared and
the respiration improved, but it was still unable to swallow solid
food and lived entirely on milk.

On the 19th of September the animal passed a solid motion, the
first since the 14th of September. As the condition appeared
improved after the injection of Novarsenobillon, a second dose of
o'og gms. was administered on the 20th of September.

No change occurred until the 23rd of September, when general
fibrillary twitchings affecting all the muscles of the body were
noticed; these twitchings were controlled by an injection of
one-eighth grain of morphia. They recurred on the 24th of
September, on which day the animal died.

A post-mortem was performed almost immediately after death.

The liver was pale yellowish in colour, and on section showed
marked fatty degeneration.

Central nervous system. ‘The cerebro-spinal fluid was slightly
turbid and contained a few polynuclear leucocytes. The surface of
the brain and cord were congested. Pieces of the cord from the
mid-dorsal and upper cervical regions, and pieces of the medulla
and cerebral cortex from the motor region (upper and lower lmb
centres), were fixed in alcohol, embedded in paraffin and sectioned ;
others were sectioned without embedding. Sections were stained in
toluidine and thionin blue and differentiated with alcohol. Eosin
and methylene blue, Giemsa, Leishman and Ehrlich’s haematoxylin
were also used.

Microscopically, the following changes were noticed in the
central nervous system :—

Many of the cells in the cord, medulla and cortex were normal
and showed Nissl’s granules. In the anterior and_ posterior
horns, and in Clarke’s column, a number of cells showed faintly
staining protoplasm and absence of Nissl’s granules, and the nucleus
tended to be eccentric in some cells. Vacuolisation of the cell
protoplasm was observed in a number of cells; the vacuoles varied
in size from 2 to 6”, and from one to six were found in each cell.
Single vacuoles were found in cells which did not show marked

|
|

301

degeneration, but were noted in large numbers only in cells where
degeneration was advanced. ‘The vessels were congested and small
haemorrhages were found. Similar changes were noted in the
medulla, where vacuolisation of degenerated nerve cells was more
marked than in the spinal cord.

Sections of the motor cortex from the upper and lower limb
centres showed engorgement of the capillaries. No haemorrhages
were found and the gross cellular changes found in the medulla and
cord were not seen.

Sections of peripheral nerves showed no pathological changes.

Cultures of the heart’s blood were negative.

On the 18th of September 0'2 c.c. of the animal’s blood were
injected intraperitoneally into a Cercopithecus campbellt with
negative result.

On the 24th of September, during the post-mortem, 3°5 c.c.
of cerebro-spinal fluid were injected intraperitoneally into a
Cercopithecus campbelli. No paralysis followed. The animal died
on the 27th of October, 1922. Post-mortem examination revealed
an abscess involving the whole of the upper lobe of the right
lung. Smears showed the presence of a Gram-negative capsulated
pneumobacillus and a Gram-negative coccus, which was isolated in
pure culture.

Before the chimpanzee’s illness it had shared a cage with three
younger chimpanzees which remained healthy. ‘This, in conjunction
with the fact that injection of blood and cerebro-spinal fluid into
Cercopithecus campbell produced no paralysis, indicates that the
condition was not one of acute anterior polyomyelitis.

The lapse of time between the administration of chloroform
and the appearance of symptoms also indicates that delayed
chloroform poisoning was not responsible for the condition.
Professor Blacklock suggests that the arsenic administered may have
contributed to the condition of the liver.

The case presents interest in its close resemblance to the course
of acute ascending paralysis as described in human beings.

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STUDIES IN THE TREATMENT OF
MALARIA—XXXII

SUMMARY OF STUDIES I—xXXxXI

J. W. W. STEPHENS
(Received for publication 25 /une, 1923)

The results recorded in the present paper constitute a summary of
work on the treatment of malaria carried out at the Liverpool School
of Tropical Medicine in the years 1917—I9QIQ.

Before considering the various treatments employed, it will be necessary
to define certain terms and it will be convenient also to consider certain
facts which emerged as the work progressed.

PART I

MALARIA

Only those cases were treated in which parasites were present in the
blood at the commencement of treatment. The patient’s temperature
may, or may not, have been above normal.

RELAPSE

A parasitic relapse, febrile or afebrile, i.e., parasites have reappeared
in the blood after a negative period induced by treatment.

FEBRILE ATTACKS

A rise of temperature above 100°F., unaccompanied by parasites in
the blood within 2-3 days, of which the nature is unknown.

OBSERVATION PERIOD

If we desire to know whether a treatment has cured a patient, i.e.,
eliminated parasites from his system, it is obvious that the patient must
be kept under observation after the treatment. The longer the patient

304

is kept under observation—by this we mean, not solely clinical observation,
but primarily (daily) microscopic examinations of the blood for parasites,
the more reasonable will it be to conclude, if the examinations are
negative—that he is cured.

The period of observation employed by us was one of 60 days, implying,
as we have just said, daily blood examinations. It should be unnecessary
to add that no treatment was given during the observation period.

FALLACIOUS FIGURES

It is necessary to point out some sources of fallacy in regard to the
results of treatment, many examples of which can be found in the
literature.

1. Absence of a microscopic diagnosis of parasites in patients before
commencing treatment. Such cases may be malaria or they may not.

2. Administration of quinine during the so-called ‘ observation period’.
The figures relating to relapses are obviously worthless.

Comparison of treatments with different observation periods.

If the value of two treatments are to be compared, the cases under
each treatment must be observed for the same length of time, after the
cessation of treatment, otherwise the figures for relapses are not
comparable, and it is impossible to say which is the better treatment,
as in the following example.

TABLE I.

| Number of

of 60 days 60 days

0 0
I Ses an 100 10 80 10 10% go %

Hil a tan 100 50 fe) 50 ens cae

305

4. Composite figures obtained by summarising the results of various
treatments.
The following is an example :—
Suppose two treatments employed, A and B, and that in the A treat-
ment the relapses were I00 per cent. and that in the B treatment they

were 0; and further, suppose that 750 cases were under treatment A,
250 under treatment B, then we get the following result :—

Cases treated Relapses Percentage
Treatment A’ ..: 4. De 750 750 100
Preatment’B.., ® ar 250 O O
1000 750 75

It is correct to say that, of 1,000 cases treated, 75 per cent. relapsed.

Let us repeat the treatments and suppose that the distribution of
cases treated now happens to be as follows :—

Cases treated Relapses Percentage
Treatment A 2; a 250 250 100
Treatment B a or 750 O O
1000 250 2

It is also correct to say that, of 1,000 cases treated, 25 per cent. only
relapsed ; but the figures 25 and 75 have no veal significance. All that
is important that the figures show, is that one treatment was very good,
the other very bad.

Percentages.

GRAPH I.

Percentage of total relapses
in each 20-day period.

306

GRAPH 2.

Percentage of cases treated
which relapse in
each 20-day period.

Days after cessation of treatment.

GRAPH 3.

Percentage of cases treated
not having previously

relapsed which do so in
each 20-day period.

307

THE TIME AT WHICH RELAPSES OCCUR AFTER CESSATION OF
TREATMENT IN SIMPLE TERTIAN MALARIA

The time incidence of relapses can be considered in three ways :—

1. In reference to the relapses themselves, i.e., the percentage of the
total relapses which occur during each period of time. From an analysis
of the time of occurrence of 582 relapses, we found that about four-fifths
occur in the first 20 days after treatment, that the majority of the
remaining one-fifth occurs in the second 20-day period, 1.e., the ratio of
the number of relapses in the two periods is about 4: I.

2. In reference to the fotal cases treated, i.e., the percentage of cases
treated which relapse during each 20-day period of time. Of the cases
treated (800), about three-fifths relapse in the first 20-day period, about
one-tenth in the second 20-day period : still fewer at later periods, 1.e., the
ratio of the percentages for the two periods is 6: TI.

3. In reference to remainders, i.e., the incidence among the cases
treated less those who have previously relapsed. Of the cases treated
(800), about three-fifths relapse in the first 20 days and about one-
fourth of ‘ the remainder ’ cases in the second 20-day period. The ratios
Seeruere 12° 5 Ol" 2.41. 1:

It is possible that, if a large number of cases that had not relapsed in
60 days had been observed for much longer periods, that the values we
have given for the first and second 20-day periods would have to be
somewhat reduced, but until the actual observations are made, this is
purely conjectural.

It must be added that, unless a sufficiently large number of cases are
considered, it is not likely that the ratios given above will be observed.

TIME OF ONSET OF THE PAROXYSMS IN SIMPLE TERTIAN
MALARIA
From an analysis of 1,000 ‘ rigors’ or paroxysms, we found that :—
(a) Over go per cent. of the paroxysms occur during the hours of
bodily activity, in our series of cases from 7 a.m. to 6.59 p.m.
(bo) The maximum number of paroxysms, about 20 per cent., occurs

at 2 p.m.
THE EFFECT OF SEASON ON TREATMENT OF MALARIA
Two series of cases consisting of 76 and 89 patients, respectively, were

treated by us at different times of the year with the same treatment,
viz., quinine sulphate, grains 90, on two consecutive days only.

308

In one series the number of relapses was about 40 per cent., in the

other, over go per cent.

The only factor that we could discover that might account for this
difference was that the cases were treated at different times of the year,
the good result was obtained in the summer and autumn, the bad result

in the winter and spring months.

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ii
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ia Ba
f is
ra rf

Grapu.—Showing the time incidence of 1,000 simple tertian malaria paroxysms ;

> =
> > a) > > e

‘Summer’ time in operation.

THE MAXIMUM DOSE OF QUININE THAT CAN BE TOLERATED

Quinine sulphate, orally in doses of grains 120 on each of two
consecutive days, represents the maximum amount of the drug which
can be tolerated by the average case, as the treatment had to be abandoned
owing to severe symptoms in five of fifteen cases.

rete

309
PART. Lh

TREATMENT OF AN ATTACK

QUININE
(a) Orally.

Ten grains of quinine sulphate in solution on each of two consecutive
days suffice to cut short an attack of simple tertian malaria, and to cause
the temporary disappearance of parasites from the cutaneous blood.

While this is so, our routine procedure is to give grains 15, two to three
times a day for a few days until the same result is accomplished. The
subsequent treatment will be considered later.

(b) Intramuscularly.
1. Quinine bihydrochloride.

Fifteen grains of quinine bihydrochloride in 2 c.c. of water on each of
two consecutive days likewise cause the cessation of febrile paroxysms and
effect the temporary disappearance of all stages of the parasites’ from
the cutaneous blood. This holds good for P. vivax and P. falciparum.

2. Quinine alkaloid.
Grains 15 to 30 in I c.c of sesame oil on each of two consecutive days,
has the same effect in cases of simple tertian malaria.
Where the patient can take quinine by the mouth there is usually
no necessity for intramuscular injections, but where oral quinine is
ineffective, intramuscular quinine remains as a most effective treatment.

Quinine bihydrochloride in doses of I0-I5 grains in a Io per cent.
solution in normal saline, in one or a series of six injections, causes the
cessation of febrile paroxysms and a disappearance of parasites from the
cutaneous blood in simple tertian malaria.

In malignant tertian malaria these doses do not cause the disappearance
of parasites—trophozoites or gametes—from the cutaneous blood.

ARSENIC
(a) Organic. Arsenobillon.

A single intravenous injection, 0.9 gramme, controls the fever, causes
the disappearance of P. vivax from the cutaneous blood within 24 hours.
The same dose has no appreciable effect on the temperature or the parasites
in the case of P. falciparum or P. malaria.

310

(b) Inorganic. Liguor arsenicalss.

In doses of 115 daily, failed to control the fever or to cause the disap-
pearance of parasites. In doses of 130 daily, the temperature fell to
normal within ten days and in 13 of 14 cases parasites disappeared in
two to six days.

SILVER ARSENIC AND ANTIMONY
Luargol.

A single intravenous injection of 0.2 gramme, controls the symptoms
and causes the disappearance of the parasites in simple tertian malaria.

ANTIMONY
Tartar Emetic.

Intravenous injections of tartar emetic, 2 per cent. solution in one
or more doses of 5-15 centigrams, do not control either the rigors or the
fever of acute malaria, nor do they cause the disappearance from the blood
of any stage of the malaria parasites, whether of P. vivax or P. falciparum.

MANGANESE
Collosol manganese I c.c. on each of two consecutive days proved
to be valueless.

QUITENINE AND QUINOTOXIN

The hydrochlorides of these derivatives of quinine proved of no value
in the doses used, viz., of about the same amount as that of quinine sulphate
which proved effective.

AMYLOPSIN AND TRYPSIN

‘ Injectio amylopsini’ and ‘ Injectio trypsini’ proved to be of no
value in the treatment of simple tertian malaria.

311

PART T(t

SUBSEQUENT TREATMENT

We have seen that the immediate effect of quinine and other drugs
is to allay the febrile symptoms and to cause the disappearance of parasites,
but this condition of apparent cure was, sooner or later, followed by a
relapse in the majority of cases. Two questions consequently arose :—

1. The first was, could the condition of apparent cure be maintained
by continuing the quinine treatment, and if so, how should it be given ?

2. The second was, were these cases in which the administration
of quinine was continued for more or less long periods, and which showed
no symptoms while taking quinine, really cured? Would they relapse
or not, when treatment was stopped, just as they had done when the
treatment had lasted only a few days, or would the number of relapses
be now smaller

QUESTION I.

The aspect of the problem that mainly occupied us was, whether if
a certain total dose of quinine were given weekly, e.g., grains 30, 60, 90,
it were better to administer the quantity on 6 days giving 5, 10, or I5 grains
daily, or on two consecutive days only each week, giving 15, 30, or 45 grains
daily.

This question was put to the test for a period of eight weeks in a series
of cases for each total weekly dose of 30, 60, and go grains of quinine
sulphate.

An accurate record was kept of the febrile relapses (non-parasitic)
and of the parasitic relapses (febrile and afebrile), as determined by the
temperature chart and daily blood examinations during the whole of
the period.

In each series the record was in favour of the weekly administration
of quinine in preference to the daily.

Thus, 30 grains is better administered in the form of two doses of
15 grains, than in the form of six doses of 5 grains.

The best result was obtained by the administration of grains 45 (three
doses of grains 15), on each of two consecutive days weekly, this as above
stated, giving a better result than grains 15 daily for six days.

An interrupted treatment of 30 grains on each of two consecutive
days weekly, also suffices to keep the blood free from trophozoites and
to prevent relapses in the majority of cases (while the treatment lasts).

312

In other words, in order to maintain a patient in a condition of freedom
from relapses, an interrupted course of quinine is preferable to a continuous
one.

So far as the actual result was concerned, an equally good one, or
nearly so, was obtained in a different way, viz., by giving 15 grains of
bihydrochloride intramuscularly on each of the first two days of treatment,
and then Liquor arsenicalis 130 daily, with two periods of intermission
for eight weeks (two weeks on, one week off, two weeks on, one week off,
two weeks on).

The comparative figures for this and the previous interrupted quinine
treatment are the following :—

Quinine injections, Quinine sulphate
two only, followed Gr. 45 on two
by Lig. arsenicalis consecutive days

I1\30 daily weekly for 8 weeks
Percentage of parasitic febrile relapse cases per
cases treated (average per week) 2°7 1-8
Percentage of all febrile (parasitic and non-
parasitic) cases per cases treated (average
per week Ms aa ves uy 43 8:7 10°3
Number of cases treated... sa8 At ee ae 74

What we have just considered is a method of maintaining freedom
from relapses while the treatment 1s in force. We shall now consider a
different question, viz. :—

QUESTION 2.

This question resolves itself into an enquiry as to whether by any
course of treatment, short or long, a curative effect would be obtained,
i.e., freedom from relapses after cessation of treatment, over an observation
period of sixty days (or longer).

Many methods were tried, but in nearly all, when treatment was
stopped, the number of relapses was large, and there is at present no method
known which will cure all cases, even if the treatment lasts eight weeks.

Many methods of cure continue however, to be advocated, but they
are not supported by trustworthy evidence, more especially in regard
to an adequate observation period.

tk Maa

:
;
4

shes

The following two treatments gave us the best results :-—

Number of
cases
treated

Liquor arsenicalis
minims 30 daily
with I or 2
periods of inter-
mission with an
injection of
quinine bihydro-
chloride on
each of the
first two days

only J

Pawetr32

Novarsenobillon — |
O-9 grm. intra-
venously on the
Ist, 8th, and
15th days with
quinine bi-
hydrochloride
gts. 15 intra-
muscularly on
the Ist and 2nd,
8th and oth,
and 16th days.

.
|
|

Number of
casesnot Number
relapsing _ of cases
but lost relapsing
sight of in an Relapses Time of Year.
before the observation per cent.
expiration period of
of 60 days 60days min. max.
( End of treatment
August :—I case ;
September :—
Pil A Cases;
October :—14 cases
— 4 L255 Dera Note.—

One additional
case was not
controlled by

the treatment.

—~

8-3 16-6 -| End of treatment
December :—I2 cases

It is worthy of note that a treatment which is ‘ good’ whilst it lasts

is not necessarily followed by a ‘ good’ result when it has ceased. Thus
the treatment noted above, viz., grains 45 x 2 weekly for eight weeks,
while ‘ excellent ’ while it lasted, was followed by 80 per cent. of relapses
when the treatment had finished.

Whereas the arsenic treatment also a good one while it lasted, was
followed by a ‘ good’ result also when it had ceased.

314

THE DISAPPEARANCE OF MALIGNANT TERTIAN GAMETES
(CRESCENTS) UNDER QUININE TREATMENT

t. With a dose of grains 30 or 45 daily, crescents do not persist in
the blood in the majority of cases for more than three weeks. Whether
they would disappear equally rapidly without quinine we did not determine.

2. Similarly with quinine sulphate grains 30 on each of two consecutive
days weekly for five weeks, the crescents diminished from 50 per cent.
in the first week to 6 per cent. in the fifth week of treatment.

315

APPENDIX

The Titles, number of volume and date of Publication of the Studies
in the Treatment of Malaria (I-NXXI) are given below :—

XVI

XVII

XVIII.

XIX

XXII

ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY.

Intravenous Injections of Tartar Emetic. Vol. XI, p. 91. 1917.

Intramuscular Injections of Quinine Bihydrochloride in Simple Tertian Malaria. Vol. XI,
Oy 113." 5017.

Intravenous Injections of Quinine Bihydrochloride. Vol. XI, p. 149. 1917.

Intramuscular Injections of Anylopsin and Trypsin in Simple Tertian Malaria. Vol. XI,
p- 165. 1917.

Intramuscular Injections of Quinine Alkaloid in Simple Tertian Malaria. Vol. XI, p. 173.
1917.

Oral Administration of Quinine for ‘Two Consecutive Days only in Simple Tertian Malaria.
Vol. XI, p. 283. 1918.

Oral Administration of Quinine Sulphate daily over Prolonged Periods in Simple Tertian
Malaria. Vol. XI, p. 309. 1918.

Oral Administration of Quinine Sulphate for Two Consecutive Days Weekly over Prolonged
Periods in Simple Tertian Malaria. Vol. XI, p. 331. 1918.

A Comparison of the Results of Interrupted and Continuous Quinine Treatment. Vol. XI,
pvjsg. 1918.

Oral Administration of Quinine Sulphate Grains 120 on Two Consecutive Days only in Simple
Tertian. Vol. XI, p. 417. 1918.

Oral Administration of Quinine Sulphate Grains 90 on Two Consecutive Days Weekly
over a Period of Three Weeks in Simple Tertian Malaria. Vol. XI, p. 421. 1918.

At what Time after Cessation of Quinine Treatment do Relapses occur in Simple Tertian
Malaria? Vol. XI, p. 425. 10918.

Oral Administration of Quinine Sulphate Grains 90 on Two Consecutive Days only in
Simple Tertian Malaria (Second Series). Vol. XII, p. 71. 1918.

Quinine Bihydrochloride Grains 30 intramuscularly, and Quinine Hydrochloride Grains 30
orally, Daily for 12 days, in Simple Tertian Malaria Vol. XII, p. 197. 1918.

A Factor hitherto overlooked in the Estimation of the Curative Value of Treatments of
Malaria. Vol. XII, p. 201. 1918.

Intravenous Injections of Novarsenobillon in Simple Tertian Malaria. Vol. XII, p. 211.
1918.

Oral Administration of Quinotoxin for two Consecutive Days only, in Simple Tertian
Malaria. Vol. XII, p. 217. 1918.

A Comparison of the Value of Continuous and Interrupted Quinine Administration in
Simple Tertian Malaria (Second Communication). Vol. XII, p. 303. 1919.

Intravenous Injections of Disodoluargol in Simple Tertian Malaria. Vol. XII, p. 339.
191g.

Intramuscular Injections of Collosol Manganese in Simple Tertian Malaria. Vol. XII,
P- 345, 919.

Arsenic in Simple Tertian Malaria. Vol. XII, p. 371. 1919.

Intramuscular Injections of Quinine Bihydrochloride Grains 15 on each of two Consecutive
Days only, in Malignant Tertian Malaria. Vol. XIII, p. 63. 1919.

XXIII

XXIV
XXV
XXVI
XXVII

XXVIII
XXLX

316

Oral Administration of Quinine Sulphate Grains 30 on each of two Consecutive Days
weekly, over a Period of Five Weeks in Malignant ‘Tertian Malaria. Vol. XII, p. 69.
191g.

The Disappearance of Crescents under Quinine Treatment. Vol. XIII, p. 73. 1919

Arsenic in Malignant Tertian Malaria. Vol. XIII, p. 75. 1919.

The Action of Arsenic and of Quinine on Quartan Malaria. Vol. XIII, p. 97. 1919.

Intravenous Injections of Novarsenobillon and Intramuscular Injections of Quinine
Bihydrochloride in Simple Tertian Malaria. Vol. XIII, p. 101. 1919.

Quitenine Hydrochloride in Simple Tertian Malaria. Vol. XIII, p. 117. 1919.

Oral Administration of Liquor Arsenicalis Minims 30 daily for 16 Days with Quinine
Bihydrochloride Grains 15 Intramuscularly on the 1st and 2nd, 8th and gth, 15th
and 16th days, in Simple Tertian Malaria. Vol. XIII, p. 119. 1919.

At what time after Cessation of Quinine Treatment do Relapses occur in Simple Tertian
Malaria ? (Second Communication). Vol. XIII, p. 125. 1919.

The Time of Onset of the Paroxysms in Simple Tertian Malaria. Vol. XIV, p. 365. 1921.

317

NOTES ON AUSTRALIAN CESTODES

P. A. MAPLESTONE

AND

eee SOE LEV ECL be
(Received for publication 3 July, 1923)
No. VII.

In this paper, which is the last of the series, three new species
and one new genus are described. Further information relating to
the morphological characters of Monopylidium macracanthum and
Linstowia echidnae are also included.

Bothridium ornatum, n. sp.

On several occasions specimens of this worm were obtained from
Carpet Snakes (Python spilotes var. variegatus, Gray), taken in the

Townsville district.

EXTERNAL ANATOMY.

The largest worm measured about 65 cm. in length, and the
greatest breadth was 7 mm.

Head. The head measures about 4 mm. in breadth and 5 mm.
in length. It consists of two cylindrical muscular tubes, one lying
dorsally and the other ventrally. They are attached to each other
throughout their whole length by a broad membrane. ‘They are
funnel-shaped and are open at both ends, the posterior weet as
being the smaller and directed inwardly (fig. 1).

Fic. 1. Bothridium ornatum,n.sp. Head. x 9.

318

Unlike B. pithonis, the anterior opening is surrounded by a very
conspicuous fleshy frill.

Segments. These are very numerous, and are all broader than
long. ‘The lateral borders are imbricated.

INTERNAL ANATOMY.

No points of difference could be observed between this worm and
B. pithonis, as described by Braun (1900).

DIAGNOSIS.

We have compared our specimens with the literature of all the
known species of Bothrvidium and find that it differs from them in
the characters of the head, viz., the possession of a fleshy frill round
the anterior openings. It is proposed to name it Bothridium
ornatum, nN. sp.

Type specimens are in the Museum of the Liverpool School of
Tropical Medicine.

Monopylidium fieldingi, sp. nov.
These cestodes were found in the intestine of several specimens
of the Butcher bird (C7vacticus destructor, Temm.), all of which were
shot in the neighbourhood of Townsville, North Queensland.

EXTERNAL ANATOMY.

A complete specimen of the worm was not available for
examination, so the total length cannot be given, but, from the
appearance of several fragments taken together, and the rate of
development, it is estimated that a complete adult would be over
50 mm. in length. The maximum breadth attained is 1°2 mm.

Head. The rostellum is strongly retracted in all the scolices
available for study, with the result that anteriorly, it is in the shape
of a truncated cone. The rostellum apparently invaginates when in
this state, so that the tip is in the form of a saucer-shaped depression,
around the edge of which is a double crown of alternating hooks
(fig. 2). The hooks are about forty in number in each row and are
of a definite rose-thorn shape when seen in profile; when viewed
dorso-ventrally they present a Y-shaped appearance, the handle of
which is long and the limbs of unequal length. They measure.about

22m in length (fig. 3).

319

The scolex reaches its maximum breadth (about 0°4 mm.) across
the posterior borders of the suckers. These organs, when viewed in
profile, are seen to stand out slightly from the surface. They are
circular in shape, and measure about 130m in diameter. They look
outwards and slightly forwards, and are unarmed.

Immediately behind the suckers the scolex narrows slightly, and
its termination is marked by a somewhat indefinite constriction
(probably an artifact), which hes about 0'4 mm. from the anterior
end. Immediately behind this constriction is an unsegmented
portion about 0'4 mm. in length and 0°25 mm. in breadth. At this
point, z.e., about o'8 mm. from the anterior extremity, the first
traces of segmentation appear.

Fic. 3. Monopylidium
fieldingi, n.sp. Hook, highly
magnified.

“BHM
Fic. 2. Monopylidium
freldingt,<n.sp.
Head. xX 35.

Segments. Segmentation soon becomes quite distinct, and the
proglottides are seen to have their lateral borders curved, with the
convexity outwards, and the postero-lateral angles projecting fairly
widely. After about the fiftieth proglottis the length increases
slightly more rapidly than the breadth, so that the proportion of
breadth to length alters somewhat, but never to such an extent that
the length becomes greater than the breadth.

Mature proglottides measure 0°17 mm. _ antero-posteriorly,
0°'4 mm. across the anterior, and 0'47 mm. across the posterior
border. The medullary portion at this stage is only 0'170 mm. in
breadth.

INTERNAL ANATOMY.
The longitudinal muscle consists of relatively thick fibres, which
are disposed in two layers, but, as the material was not in a good

320

enough state of preservation, and was also somewhat scanty, sections
could not be cut, and therefore a detailed description of the
musculature cannot be given.

Nervous system. ‘The details of this system were not investi-
gated.

Excretory system. he longitudinal excretory vessels lie at a
considerable distance from the lateral borders, so that the medulla
is correspondingly narrow. The ventral vessels are uniform in
diameter throughout their whole length; they measure about 20 in
optical section. ‘The narrower dorsal vessel lies directly above the
ventral, and the ducts from the reproductive organs pass between
them.

Genitalia. ‘The genitalia develop slowly, so that there are about
one hundred segments showing traces of the sexual organs before
they become sufficiently developed to be clearly distinguished.

Testes. The testes are circular or slightly oval, and number
about sixteen to twenty-one in each segment. When viewed dorso-
ventrally they are seen to occupy the space posterior to the female
glands, but on each side a few follicles are on a level with the
vitellarium, or even with the ovary itself. The vasa efferentia unite
into a vas deferens, which is thrown into several coils in front of the
right lobe of the ovary. There is no external vesicula seminalis, and
the vas deferens passes directly into the base of the cirrus pouch.
The cirrus pouch is relatively long and narrow, its dimensions being
130# in length and 45m in breadth. Beginning mesial to the
excretory vessels, it runs towards the right side in all cases, and very
slightly posteriorly, and, passing between them, opens in a small
atrium, which in turn opens on the right lateral border, about the
junction of the anterior and middle thirds. The characters of the
cirrus could not be made out (fig. 4).

Receptaculum and vagina. The vagina is a long straight tube
which commences at the genital pore, immediately posterior to the
opening of the cirrus. From here it runs transversely inwards, thus
diverging more and more from the cirrus pouch as it goes; it passes
dorsal to the right lobe of the ovary, dilating over the ovarian duct
into a small but distinct receptaculum seminis.

Ovary. The ovary is centrally situated in the anterior half of
the medulla. It is approximately bilaterally symmetrical and

321

consists of three lobes, two pointing laterally and a median lobe
pointing anteriorly (fig. 4).

Uterus. he uterus develops as a uniform sac devoid of out-
pocketings. It eventually fills the entire segment antero-posteriorly
and extends laterally to the excretory canals.

Fic. 4. Monopylidium fielding, n.sp. Ripe segment, showing genitalia. x 69.

It is split up into capsules having a reticular form, each capsule
containing up to about twelve eggs. Later, a separate capsule
appears to be formed around each egg. ‘The uterus was not fully

Fic. 5. Monopylidium fieldingi, n.sp. Gravid segment. x28.

matured in any specimen, therefore the nature and extent of these
capsules could not be determined (fig. 5).
Eggs. No ripe eggs were seen.

$22

DIAGNOSIS.

As this worm possesses all the characters given by Fuhrmann
(1899) in the diagnosis of Monopylidium, there is no doubt that it
belongs to this genus.

As it disagrees with all known species of Monopylidium, it is
consequently new and accordingly named Monopylidium fieldingi
after Mr. J. W. Fielding, Senior Assistant at the Australian Institute
of Tropical Medicine, who collected this and most of the other
material described in this series.

Type specimens are in the Museum of the Liverpool School of
Tropical Medicine.

Monopylidium macracanthum, Fuhrm.

This worm was found on several occasions in the intestine of the
spur-winged plover (Lodivanellus lobatus, Lath.).

EXTERNAL ANATOMY.

Fixed specimens measure about 45 mm. in length, and 1°8 mm.
in breadth, but there is apparently some shrinkage from fixation, so
that these dimensions will have to be revised when fresh material 1s
available.

There is no trace of a ‘neck’, the scolex passing directly into the
segmented chain.

Head. The scolex is about 06 mm. in breadth, and is conical
anteriorly (fig. 6). The suckers are relatively small, but well

>
26,

E.H.M.

Fic. 6. Monopylidium macracanthum, Fuhr. Head. X 35.

323

developed ; they measure 0°13 mm. in diameter, and are situated on
the scolex just where it begins to narrow anteriorly. They face
forwards and slightly outwards. The tip of the scolex bears a
rostellum, which was only seen in the retracted state in all the
specimens available for examination.

Rostellum. The rostellum is a muscular organ, and completely
fills the fossa into which it is contracted ; this fossa measures about
0°47 mm. in depth and o'24 mm. in greatest breadth, being oval in
shape. The rostellum is seen to consist of two parts, a small anterior
conical part and a larger oval posterior portion. The anterior part
is distinctly marked off from the posterior by a neck-like constriction.
It is armed with a double row of relatively large hooks, which
number twenty-six (? twenty-eight). They are of two sizes,
alternating, the larger measure 145“ and the smaller about 11op.
They have a long dorsal root and blade, and a-short ventral root,
145m (fig. 6).

The posterior portion of the rostellum is oval in optical section,
and, in the contracted state, appears very muscular.

Segmenis. The dimensions of sexually mature segments are
about I mm. across the anterior, and 1°2 mm. across the posterior
borders, so that the postero-lateral angles are only slightly
projecting ; their length is 0°4 mm., giving a proportion of breadth
to length of approximately three to one (fig. 7).

INTERNAL ANATOMY.

Muscular system. ‘The cuticle is thickly studded with calcareous
corpuscles, and the muscle layers are only thinly developed, but their
exact disposition cannot be given as there was not sufficient material
from which to cut sections.

Nervous system. ‘This was not carefully investigated. It was
noted, however, that a single nerve ran external to the excretory
vessels.

Excretory system. ‘The lateral excretory vessels are situated
directly one above the other, the dorsal being the narrower. The
ventral vessels are joined by a commisural channel, which runs across
immediately posterior to the testes.

Genitalia. The reproductive organs can first be made out in
about the tenth proglottis, and from here on they steadily become
more distinct, until they reach maturity. There are about fifty

324

proglottides sexually mature as far as microscopic characters go,
before the uterus becomes apparent.

The genital pores are circular, relatively large and irregularly
alternating.

Fic. 7. Monopylidium macracanthum, Fuhr. Segments showing genitalia. x 35.

Testes. The testes number twenty-three to thirty in each
segment, and they occupy the usual dorsal position lying on each
side of, and posterior to, the ovary when viewed from above
(fig. 7).

Vas deferens. The various vasa efferentia pass forwards and
unite in front of the ovary into a many-coiled vas deferens, which
apparently fulfils the function of a vesicula seminalis, as there is no
indication of this organ otherwise. These coils lie transversely on
the pore side about the junction of the anterior and middle thirds
of the segment, and run direct into the mesial end of the cirrus sac.
The cirrus pouch is long and thin, and contains a few coils of
vas deferens and the cirrus. In its course it runs laterally between
the excretory vessels, and, passing slightly posteriorly, opens about
the middle of the lateral border of the segment. The characters of
the cirrus could not be clearly determined, as it was in all cases
entirely within the pouch.

Ovary. The ovaries consist of two equal lobes placed one on
each side of the mid-line, and each is composed of many subsidiary
branches (fig. 7). The anterior border of the ovary is about the

325

level of the mid-transverse plane of the segment. The lobes each
measure about 80m in diameter. A duct runs inwards from each
lobe and they unite in the mid-line to form the oviduct which joins
the fertilisation canal running to the uterus. From this junction a
duct passes directly backwards to the vitellarium, and it is
surrounded for a little part of its length by the small compact shell
gland.

Receptaculum and vagina. ‘The vagina opens from the genital
atrium, ventral and posterior to the male opening. It is a straight
tube which at first follows the posterior border of the cirrus pouch
as it passes mesially, but it soon leaves this and runs directly
inwards towards the anterior border of the ovary, where it dilates
in front of the lobe on the pore side, into a small receptaculum
seminis; from the mesial end of the receptaculum a duct passes
posteriorly to join with the two branches of the ovarian duct (one
from each lobe), where they unite.

Vitelline glands. ‘The vitellarium les behind the ovary in the
mid-line. It 1s a compact body, showing no trace of branching, but
is somewhat indefinitely divided into right and left lobes, with the
result that the whole organ is more or less kidney-shaped, with the
‘hilum’ facing forwards. A duct runs from its centre anteriorly,
to join the fertilisation canal.

Uterus and eggs. The uterus is at first simple, and saccular,
but later it splits up into capsules, each containing a single onco-
sphere. The eggs are circular or slightly oval, being about 80m in
diameter; the contained embryo measures about 30p.

The male copulatory organs persist and are quite distinct even
in fully gravid segments, long after all the other reproductive organs
have entirely disappeared.

DIAGNOSIS.

The presence of a double crown of hooks, identical in size with
those of Monopylidium macracanthum, Fuhrmann, together with
the uterus split up into capsules each containing one oncosphere,
leaves no room for doubt that this species 1s M7. macracanthum,
Fuhrmann. The only point in which it differs is in the number of
hooks. Fuhrmann gives twenty-two, whereas in the present species
at least twenty-six were seen clearly. As hooks easily become

326

detached, and further, as their number often varies, no importance
can be placed on this difference. -

Fuhrmann (1907) originally recorded this parasite from
Helodroncus octropus in Africa and India, and as his description
is somewhat meagre, it was thought desirable to amplify his account
when making this new record of the worm in a fresh host and
locality, viz., Lobivanellus lobatus, from North Queensland.

Type specimens of this cestode were placed in the Museum of the
Liverpool School of Tropical Medicine.

Linstowia echtidnae, Thompson (1893).
D’Arcy Thompson (1893) described a cestode from the Echidna

from Australia. In his brief description he mentions that the worms
were very contracted.

We have a large collection of immature worms from the same
host which, as far as can be ascertained, are the same species.
Our material, however, 1s not so strongly contracted as that of
Thompson, and accordingly the condition of the scolex in particular
is somewhat different.

As Thompson’s description is rather incomplete, the following
additional particulars are given.

Head. The anterior surface of the scolex is quite devoid of a
rostellum, in fact in some cases it has a slight central depression.

The dimensions of the rounded scolex differ shghtly in different
specimens, varying between 0°76 mm. and 0°58 mm. in breadth.
The maximum diameter is just posterior to the suckers.

Suckers. The four suckers are placed well forward on the scolex ;
they are well developed, circular organs, lying flat on the surface,
and their openings look outwards and slightly forwards (fig. 8).

Segments. At first the proglottides are almost rectangular in
shape, broader than long, with no projection of the postero-lateral
angles; but as development advances the posterior angles come to
project somewhat, with the result that the anterior borders of the
segments are shorter than the posterior. The dimensions of the most
fully developed segments available for study are 1°6 mm. across the
posterior borders, and about 1°35 mm. across the anterior, with a
length of about 0°45 mm., being approximately a proportion of
breadth to length of three to one, The posterior border is slightly

327

curved, with the convexity backwards, and to some extent it
overlaps the succeeding segment. The cuticle of the worm is thrown
into several slightly marked longitudinal folds, which on the

Fic. 8. Linstowta echidnae (Vhompson). Head and anterior segments. X 12.

posterior free borders of the segments give an appearance of

scalloping (figs. 8 and Io).

INTERNAL ANATOMY.

Muscular system. Transverse sections show a relatively thick
cuticle and cortical parenchyma, and the longitudinal muscle is
disposed in two layers, completely encircling the segment, the outer

layer being slightly the thicker of the two (fig. 9).

Fic. 9. Linstowia echidnae (Thompson). ‘Transverse section showing musculature. x 35.

Nervous system. This system was not investigated.

Lxcretory system. The dorsal excretory vessel is narrower than
the ventral, and lies to the outer side of the latter. They both pass
dorsal to the ducts of the male and female organs.

328

Genitalia. The genital pores cannot be made out, as in no
instance is development complete enough to show them, but from the
direction of the immature sex ducts they would probably open about
the centre of the lateral border; they are irregularly alternating,
there being, as a rule, three or four on one side followed by about
the same number on the other side. The reproductive organs are
single in each proglottis (fig. 10).

Fic. 10. Linstowta echidnae (Thompson). Segments showing cirrus pouch and vagina. xX 35.

Testes. Owing to the immature condition of the worms, only
traces of the testes.can be distinguished. [hey are numerous and
are scattered dorsally across the whole width of the medulla.

Vas deferens. No details of this organ can be made out, but it
is seen entering the mesial end of the developing cirrus pouch. At
the most advanced stage of development observed, the cirrus pouch
is represented by a relatively long, straight, tubular structure, which
runs inwards and slightly forwards from opposite the centre of the
lateral border, so that it hes across the antero-lateral angle of the
segment on the side on which it will eventually open.

Ovary. This organ lies slightly to the pore side of the median
line, and about midway between the anterior and posterior borders
of the segment. No details of its structure can be given because it
is quite Immature.

Receptaculum and vagina. The vagina is seen as a straight
tube, running inwards along the posterior border of the cirrus pouch,
which it leaves about its centre, and running directly inwards, ends
in a small expansion, evidently the beginnings of a receptaculum
seminis, around which the developing female genitalia can be seen.

329

On account of the undeveloped condition of the worms, no
particulars of the vitelline glands, shell glands, uterus or eggs can
be given.

Paramoniezia suis, n.g., Nn. sp.

One specimen was obtained from the intestine of a wild pig
(Sus scrofula), near Townsville, North IS ETELETA

EXTERNAL ANATOMY.

The worm 1s lancet-shaped and measures about 12 cms. in
length and 10 mm. maximum breadth.

Head. ‘he head is very small and measures only about 3004
in breadth. It is unarmed and there is no rostellum. The suckers
are extremely small; they were too shrunken to give accurate
dimensions. There is no neck.

Segments. These are always broader than long and their free
edges are imbricated. A typical mature segment measures 200m 1n
length and 9 mm. breadth. The genital pores are double.

INTERNAL ANATOMY.

Muscular system. The longitudinal muscle fibres are arranged
in a single layer, composed of numerous bundles measuring about
604 thick. External to this 1s a thin layer of circular fibres. A few
dorso-ventral fibres also occur.

Nervous system. This system could not be investigated because
even the main lateral nerve could only be seen with difficulty.

Excretory system. The ventral excretory vessel is very large,

_and in most transverse sections it appears to occupy the whole of the

lateral dorso-ventral space. The diameter of the tube is about
150M.

The large size of this vessel, and the numerous branches to which
it gives rise, made it difficult to determine whether a dorsal vessel
was present or not; but careful examination led us to the conclusion
that a dorsal vessel was absent.

Genitalia.

Testes. These are very numerous (at least three hundred).
They extend on each side almost to the lateral extremity of the
segment and are not grouped round the ovary, but extend right

53°

across the segment. Each testis measures about O05 by 45x.
Antero-posteriorly they lie in four or five rows, and dorso-ventrally
in from one to three layers.

Vas deferens. The vas deferens on each side runs dorsal to the
ventral excretory vessels. The cirrus pouch is tubular and _ lies
lateral to the water vessel. Its median portion contains an internal
vesicula seminalis. No external vesicula seminalis was seen. The
cirrus 1s unarmed.

Ovary. This organ is paired in each segment, and is of the
usual Cz¢totaenta or Moniezta type.

Receptaculum and vagina. From the pore the vagina runs
inwards, having at first a diameter of about 60; it expands
immediately internal to the excretory vessels into a large transversely
elongated muscular sac (the receptaculum seminis) measuring about
650 in length and 150p breadth. Its median extremity, which
lies close to the ovary, is continued as a short coiled narrow tube
to the fertilization canal.

A most important point is the fact that whilst the vagina 1s
always ventral to the cirrus pouch on right side, it may be either
dorsal or ventral on the opposite side.

Uterus. This is first apparent as a cell-string running across
the segment. It develops into a tube, devoid of outgrowths, and
extends on each side to the extreme edge of the segment.

Eggs. The ripe egg has a diameter of 45; the outer shell has
a double contour. The hexacanth embryo measures about 24, and
a pyriform apparatus is entirely absent. Between the embryo and
the shell a small quantity of yolk can be seen.

DIAGNOSIS.

This worm obviously belongs to the family Axoplocephalidae,
Fuhrm., 1907, and the sub-family Axoplocephalinae, Blanchard,
1891. The only two genera within this sub-family possessing
double genital pores and a single cirrus pouch on each side are
Cittotaenia, Rheim., 1881, and Monzezia, Blanchard, 1891, and
these differ from each other in one particular only, viz., im
Cittotaenia the vagina is ventral to the cirrus pouch on both sides,
whilst in Monzezia the vagina is ventral to the cirrus pouch on the
right side and dorsal on the left. In the present species the
relationship of the cirrus to vagina is variable, the vagina being

ong

331

sometimes dorsal and sometimes ventral to the cirrus on the left
side, in the same strobila.

It is necessary, therefore, to erect a new genus for this species,
which we have named Paramoniezia suis, n.g., n. sp.

The characters of the new genus are as follows :—Paramoniezia.
With the characters of the genus Moniezia, except that on the left
side the cirrus is sometimes dorsal and sometimes ventral to the
vagina.

Type specimens are in the Museum of the Liverpool School of
Tropical Medicine.

REFERENCES

Braun, M. (1894-1900). Bronn’s Klassen und Ordnungen des Thier-Reichs, p. 1690. Leipzig.

Funrmann, O. (1899). Mitteilungen uber Vogeltinien. Pt. 3. Centralb. f. Bakt. etc.
I. Abt. Originale. Vol. XXVI, pp. 622-627. Jena.

——— (1907). Bekannte und neue Arten und Genera von Vogeltanien. Centralb. f. Bakt. etc.
I. Abt. Originale. Vol. XLV. Jena. pp. 516-536.

Tompson, D’Arcy, W. (1893). Note on a Tapeworm from Echidna. (Taenia echidnae
sp. n.). Journ. R. Micr. Soc., London. 1893. III, p. 297.

ZscHoxkE, F. (1899). Entozoen der aplacentelen Saiugetiere. Proc. 4 Internat. Cong. Zool.
Pp. 203-204. Lond. (Cambridge, Eng., Aug. 21-27, 1898).

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333

A NOTE ON OPHIOTAENIA PUNICA
(CHOLODOVSKI, 1908), LA RUE, ror1

BY
DEW ELL
AND
ae ABER

(Received for publication 3 July, 1923)

Iwo specimens, both gravid, were obtained from the intestine
of Causus rhombeatus, in Freetown, Sierra Leone.

EXTERNAL ANATOMY.

The larger complete specimen measured 9 cms. in length, and the
maximum breadth was 3°3 mms.

Head. The head is almost square and measures 1°5 mm. broad;
it is unarmed.

Suckers. The four suckers have a diameter of 0.67 mm. The
neck is 0'7 mm. long.

The worm is made up of about one hundred and forty segments.
The first proglottides are broader than long; they gradually lengthen
towards the posterior, the last proglottis being 4 mm. long and
2 mm. broad.

The genital pores are irregularly alternate, and open at the
middle of the lateral border.

INTERNAL ANATOMY.

Musculature. The musculature consists of a series of (1) small!
subcuticular fibres, situated immediately beneath the cuticle, (2) a
double layer of longitudinal muscles which are not strongly
developed, (3) a few diagonal fibres, and (4) circular fibres which are
very scanty.

Excretory system. There are two water vessels on each side,
the ventral vessel being much larger than the dorsal vessel.

Nervous system. A single nerve is present on each side, lying
lateral to the water vessels. The parenchyma is strongly developed.

Genitalia. The testes are confined to the lateral fields in front
of the ovary, and median to the vitellaria. There are from
one hundred and seventy to two hundred and thirty in each
segment; they are oval in shape, their long axes being horizontal

fe. 1).

334

Vas deferens. The cirrus pouch first becomes evident about
15 mm. behind the head; it lies either anterior or posterior to the
vagina and extends beyond the vitellaria, being up to 670y in
length (fig. 1).

EHM

Fic. 1. Ophbiotaenia punica. A ripe segment, shewing genitalia. v.g.—-Vitelline
glands. c.p.—Cirrus pouch. v.—vagina. t.—testes. 0.—Ovary. v.d.—Vas deferens.
u.—Uterus. “ °35<
The cirrus is spiny and is continuous with an internal seminal

vesicle, which latter occupies about two-thirds of the cirrus pouch.
The vas deferens lying outside the pouch is coiled.

Ovary. The ovary is long and narrow and is not bilobed
(fig. I); it is situated posteriorly.

Vagina. The vagina lies either anterior or posterior to the
cirrus pouch, It runs almost straight towards the middle of the
segment, and then turns posteriorly (fig. 1).

Vitellaria. The vitellaria are lateral, and consist of small acin:
measuring about 30 to 36m in diameter (fig. 1).

Uterus. The uterus is a_ straight tube running antero-
posteriorly ; in mature segments it has from eight to twelve lateral
pouches on each side. There is a small shell gland situated
immediately behind the middle of the ovary (fig. 2). In transverse

Fic. 2. Opbiotaenia punica. A segment shewing gravid uterus. v.g.—Vitelline
glands. c.p.—Cirrus pouch. v.—Vagina. «.—Uterus. 0—Ovary. X 35.

Che

sections of segments in which the uterus was gravid no uterine pores
were seen.

Eggs. Uhe eggs are 30 in diameter, and in appearance
resemble the eggs of Hymenolepis nana. The oncosphere is from
13@ to 15m“ in diameter. The embryophore has a thickness of about

3m (fig. 3).

EHM.

Fic. 3. Ophiotaenia punica. Egg. x 733.

Diagnosis. Ophiotaenia punica was first found in a dog in
Tunis by Cholodovski (1908), but, owing to its morphological
characters, Hall, Ransom and La Rue thought the true host was a
snake. They presumed that the dog had eaten a snake. Southwell
(1922) recorded this parasite from Paradoxurus hermaphroditicus
(Malayan palm civet) in Calcutta.

This is the first definite record of Ophiotaenia punica from a
snake.

REFERENCES

Cuoropovski (1908). Uber ein neuer Parasit des Hundes. Zool. Anz., Vol. XXIII.

La Ruz, G. R. (1914). A Revision of the Cestode Family Proteocephalidae. Illinois Biol.
Monog., Vol. I, Nos. 1 and 2. Urbana.

SOUTHWELL (1922). Cestodes in the Collection of the Indian Museum. Ann. Trop. Med.
and Parasit., Vol. XVI, p. 127.

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337

A PYRRHOCORID BUG CAPABLE OF
BITING MAN

ee DES Creo Kreme

From the Sir Alfred Lewis Jones Research Laboratory,
Freetown, Sierra Leone

(Received for publication 4 July, 1923)

So far as I am aware, the following notes contain the first record
of the fact that a bug of the family Pyrrhocoridae has been found
to biteman. The observation was made at Freetown, Sierra Leone,
but so near to the time of my departure that little opportunity of
carrying out experimental work on the subject was available.

The natural order Rhynchota or Hemiptera is divided into
the sub-orders Heteroptera and Homoptera; the Heteroptera into
Gymnocerata and Cryptocerata; the Gymnocerata into several
families, of which the family Pyrrhocoridae is one; in a sub-family
of this—the Pyrrhocorinae—occur many genera, to one of which,
namely, Dysdercus, belongs the insect with which I am here dealing.
Mr. Lang, of the British Museum, has kindly identified the species
for me as D. superstitiosus, F.

Dysdercus superstitiosus was observed by me at Freetown, in
1921, specimens being found even indoors in the laboratory. At
that time I made a few experiments in order to see if the insects
would bite; the bugs were placed singly in wide test-tubes and
applied to the arm, but they showed no inclination to bite or even
stay on the skin, making, on the contrary, efforts to escape by
climbing up the test-tubes; it was concluded, rather prematurely
as it now appears, that they were entirely non-biting in so far as
human beings are concerned. They were observed in numbers
on the ground, especially in the vicinity of a silk cotton tree,
Eriodendron anfractuosum, situated about a hundred yards from
the laboratory. It was observed that they appear to see very well,
as they are extremely sensitive to any movement made in their
vicinity; also that they make off instantly and move away with

338

great rapidity when disturbed. This year, in April, the silk cotton
tree pods were opening on the tree and it happened that on one or
two days the wind drove the cotton along, in the direction of the
ground on which the laboratory stands. In the flocculi of silk
cotton which were wafted on to the ground were seen numerous
small red bugs, which in most cases extricated themselves quickly
and ran about actively. Occasionally a silk cotton seed was carried
along with the floating fragments of fibre, and it was not unusual
to see on the ground a seed covered with bugs, some of them with
their beaks inserted into it and others trying to pierce: it, the seed
being pulled in all directions during ‘the process. When the bugs
grew larger some were observed to develop cannibal habits, more
especially when placed together in test-tubes with no food.

On the 24th of April, 1923, while sitting out in front of the
laboratory just after sunset I experienced a sharp bite on the front
of the ankle. On looking to see what was biting 1 moved slightly
and could observe no mosquito or other biting insect, but saw a
red bug moving rapidly off my sock. It was not possible to be sure
that the definitely painful bite was caused by the bug; the reaction
was a small itching swelling which had disappeared in twenty-four
hours. Two days later, on the 26th April, at the same place and
time, a bite was again felt on the same ankle. Leaning forward
carefully I saw a red bug biting busily through my thick black sock.
The bug’s body, as its beak went deeper and deeper into the skin,
assumed an attitude which was nearly vertical; considerable
irritation was felt at intervals during the time the biting was going
on. The process was timed and had lasted nearly four minutes
when the bug was disturbed by a large black ant, which, in passing
rapidly across the ankle, collided with the bug. The latter instantly
made off and was escaping when it was captured in a test-tube
which had been kept at hand since the previous observation, so as
to be ready should the opportunity arise again. The reaction was
on this occasion also only local, but was much more definite.
Itching and irritation were felt, and in an hour’s time a circular
swelling of the size of a sixpenny piece had developed, well raised
in the centre. This remained and in two days was rather hard,
but it went away gradually in about five days from the time of the
bite. [he bug was examined and found to be a last larval stage:

339

the dissection of it did not reveal the presence of blood in the
alimentary canal, nor were flagellates found; but the dissection
had to be carried out with the light of an oil lamp, which was
unsatisfactory.

Laboratory experiments. Several experiments were carried out
by placing single bugs in large test-tubes on the human skin, but in
no case was biting observed, the bugs being only anxious to escape
by climbing up the tubes. Too much stress should not be laid upon
the negative results of these experiments, in view of the fact that
bugs which had been seen attacking the seed of the silk cotton tree
on the ground made no attempt to attack the seeds when placed
with them in tubes similar to those used for the skin experiments.
Ballou (1906) made a comparable observation. He says: ‘Although
the cotton stainers are known to feed on the ripe cotton seed about
the gin-houses, they would not do this in the laboratory, nor would
they feed on the seeds of the silk cotton.’ It is probable that on
account of the timidity of the bug, special methods of experimenta-
tion in the laboratory must be devised.

Success, however, attended an experiment which was carried out
in England. Two bugs were placed in the toe of a black sock,
which was then drawn half on to the foot. After some minutes a
bite was felt on the dorsum of the foot, but in the attempt carefully
to remove the sock the bug was disturbed and ceased biting; the
reaction was an itching sensation with subsequent local swelling,
and for a few days a red circular area on the dorsum of the foot was
observed. Later the whole leg became swollen and oedematous, and
intense itching occurred; there was no pain, however, and no
enlargement of glands and no temperature. Parasites were not
found in the blood, but there was eosinophilia reaching 50 per cent.
In three weeks the swelling began to go down, and in a month was
gone.

The bionomics of Dysdercus. This genus of bugs is best known
from its association with cotton in most parts of the world, and it
contains the majority of insects classed as cotton stainers. Maxwell-
Lefroy gives a detailed account of the morphology and bionomics
of D. cingulatus which occurs in India, as well as notes on other
species; Ballou (1906) enumerates a large number of species which
are found in the West Indies, with observations on the various stages

340

of the life-history in different species. Egg-laying commences
within a few days after copulation is over, the eggs being laid to the
number of up to a hundred in various sites, on the ground, under
leaves, in the open bolls; Peacock, in D. superstitiosus in captivity,
found egg clusters from twenty to one hundred and twenty ; the egg-
stage lasts abcut a week, and the larva which emerges undergoes
during its growth five ecdyses. The adult is distinguished from
the larva in most cases, according to Butler (1923), by the acquisition
of wings; increase of number of joints in tarsi, and sometimes in
antennae; transference of openings of scent glands from dorsal to
ventral side; full development of sexual organs. The time taken
for growth from the time the egg is laid till the adult bug stage is
reached is forty-nine to eighty-six days for D. cingulatus, in India.

Food Plants. The chief food plants of these bugs are the
following :—The cotton Gossypium sp. ; the silk cotton, k7zodendron
anfractuosum; the Okra or Bhindi, Hibiscus esculentus; the musk
mallow, Hzdizscus abelmoschus, and other plants of the Malvaceae.

Other food. Dysdercus superstitiosus was observed by me at
Freetown feeding in large clusters on the carcase of a frog.
Mansfield-Aders (1919-20), in his account of insects injurious to
economic crops in the Protectorate of Zanzibar, states that he has
on many occasions seen Dysdercus fasciatus feeding with avidity
on fresh mammalian carcases, skins, and skulls. Of D. super-
stitiosus in Zanzibar, he says that it is by no means a common
species on cotton, but that the silk cotton is commonly attacked
by it.

Lamborn (1914-15) says of Southern Nigeria, ‘ During the dry
season the Pyrrhocorid bug, Dysdercus superstitiosus, F., was
found in some numbers . . . . and at this time they appeared to
be able to thrive on almost any food, whether of animal or vegetable
origin, for eight or ten were noticed feeding on a dead and sun-
dried lizard and a batch of young nymphs was found on sheeps’
excreta.’

Peacock (1913-14) gives a coloured plate of the stages of
D. superstitiosus. He records the observation which he made of a
number of young stainers about three weeks old sucking a dead
snail.

Seasonal occurrence. Lefroy gives the following account of the

341

sequence of breeding and feeding habits of D. cingulatus at
Pusa :—
April-May—Extensive breeding on Simul (silk cotton).
June-July—Feeding miscellaneous Bhindi, Hibiscus, etc.
August-November—Breeding in cotton.

‘In most parts of India breeding is of necessity confined either
to the cotton season, to the season when Bhindi is in pod, or to the
season when the Simul is in bearing.’ These observations are of
interest with regard to the effect of season and the presence of
particular food plants on the numbers and vitality of a single
species.

Distribution of Species. Of equal interest are the observations
made by Ballou on the distribution of different species in the West
Indies. One species may extend for a distance and then, apparently
without any change of environment, stop and give place to a different
species. This sudden demarcation of the limits of a species is most
noticeable in the case of D. rvuficollis, and it is stated ‘in many
instances only one locality is known for each species, and most of
the others occur only in a few adjoining countries or islands.’ The
distributions of D. axdreae and D. delauneyi give good examples of
localization. It is noted by Lefroy that D. evanescens, Dist., 1s
recorded from Sikkim, the Khasi and Garo hills, Burma, from the
Bor Ghat, Bombay, also from Chapra.

Modification of feeding habits. Butler (1923) considers the
possibility of rapid change of food habit among bugs. He refers to
two species of Capsidae which are even now gradually establishing,
or indeed have established, themselves in orchards, viz., Pleszocoris
rugicollis and Orthotylus marginalis; the natural food plants of
these insects are various species of Salzx, ‘and the attack upon
orchards indicates a startling change of taste brought about by the
temptation of well-nurtured plantations of apple trees in their
neighbourhood.’ In captivity Ballou fed stainers on cotton seed,
portions of unripe cotton bolls, bits of sugar cane and pieces of
banana.

Duration of Life. In the insectary D. cingulatus was kept by
Lefroy for four months; life was long when conditions were not
favourable, z.e., little food. I may observe here that a few specimens
of D. superstitiosus, last larval instar and adult, which received no

342

food and which were kept at ordinary temperature, survived the
voyage from Sierra Leone to England and lived for a week after
arrival,

Bacterial parasites of Dysdercus spp. De Charmoy (1921) says
that in Mauritius Dysdercus is known to transmit several bacterial
diseases, and that it is the vector of an internal disease of the bolls
similar to that described by Nowell and others in the West Indies.

SAAS
¢ se

| l | Qow

Fic. 1. Flagellate parasite of Dysdercus superstitiosus, F.

Flagellate parasite of Dysdercus superstitiosus, F. The bugs
were found by me to be infected with a herpetomonas; it was present
in various portions of the alimentary canal, and was recovered once
from the coelomic fluid by cutting off the antenna near its base and
examining the fluid which exuded; it was not found in the salivary
glands, but as will be seen from the table subjoined, the number of
dissections was limited :—

343

Flagellates (Herpetomonas) present in Dysdercus superstitiosus F.

| Salivary | Coelomic
Dissected Infected Rectum Hind gut Mid gut | glands | Fluid
4 9 dy 9 7 2 ° I

The occurrence of flagellates in Hemuiptera-Heteroptera is well
known. Patton and Cragg (1913) refer to the fact that tea, coffee
and garden produce of all kinds are attacked by various species of
bugs. They mention the family Lygaeidae, of which several species
are infected with flagellates; Orycarenus laetus, which 1s common
on the cotton plant in Madras, is nearly always infected with a
species of Herpetomonas; Lygaeus pandarus (militaris), which is
common on the milk plant, Calotrofis gigantea, is also infected with
a flagellate of the same kind, Herpetomonas lygaet, which very
closely resembles the parasite of Kala azar; Lygaeus hospes 1s
infected with the same parasite. Of H. lygaez, Patton, the authors
say that it is indistinguishable in its pre- and post-flagellate stages
from the parasite of Kala azar as seen in man. The observations
made by me on the flagellates of Dysdercus superstitiosus bear out
these statements as regards the appearance of the Herpetomonas
found, in the flagellate stage.

It is of interest to recall the discovery by Lafont (1909) of
Herpetomonas davidi in the latex of Huphorbia pilulifera and to the
presence of this flagellate in a species of Nysius. Miss Robertson
has recorded a herpetomonas from the alimentary tract of Dysdercus
casiatus, the red cotton bug of Uganda; only a few infected
specimens were examined. A still more important observation of
Miss Robertson is in connection with the species Lef/oglossus
membranaceus, of the family Coreidae in Uganda; she found in its
alimentary tract a herpetomonas; the parasite was found very
frequently to invade the salivary glands. Miss Robertson considers
notable ‘ the independent development in a sucking insect of all the
factors requisite for the transmission of a flagellate, parasitic in the
intestine by way of the mouth-parts of the insect host.’

The significance of the observation which I have made on the
biting capability of Dysdercus superstitiosus, F., is evident from a

344

consideration of the foregoing facts. The work of Laveran and
Franchini (1913) and of Fantham and Porter (1915) went to prove
that insect herpetomonas may, when injected into animals, produce
effects analogous to those occurring in Kala azar, the flagellates
resembling cultural forms of Lezshmania donovani, giving rise to the
non-flagellated rounded forms. The partial development in the
bed bug obtained by Patton of Kala azar parasites by feeding
experiments is important, and especially so in view of the non-
success which has hitherto attended all attempts to find an insect
vector of the parasite of this disease.

Roubaud and Franchini (1922) obtained in mice infection with
Leishmania forms of parasite by allowing fleas having a natural
infection to breed in their box. The infection, which proves fatal to
the mice, was conveyed to fresh mice by means of subcutaneous
injection of ground-up tissues. These workers also obtained a
similar result 1: mice by injecting into them the faeces of fleas.

Although there is, as yet, no evidence that Dysdercus super-
stitiosus, F., is capable of removing blood from man, there is ample
evidence that in biting it is capable of injecting an irritating
substance under the skin. This irritating substance can on analogy
be none other than the salivary fluid; it is clear, therefore, that all
the conditions for transference of a parasite to man are provided
if salivary infection is present. Dr. P. A. Maplestone reports that
he has found infection of the salivary glands. |

There is a hypothesis put forward by Stephens (1915) to explain
the lack of success of infecting arthropods from Kala azar cases.
On this hypothesis—the hemi-cyclic hypothesis—uit 1s possible that
a biting arthropod may infect man by its bite; the parasite injected
into man grows and multiplies in the tissues but does not enter the
peripheral blood in sufficient numbers to cause an infection in the
alimentary tract of a fresh arthropod when biting; this is tantamount
to saying that the parasite which gets into the arthropod from some
other source than man reaches in man a cul de sac from which it
cannot escape. The hemi-cyclic hypothesis, however capable it
might be of explaining transmission of disease by the bites of insects
which were yet not capable of sucking blood, need hardly be
considered here, as we do not know what are the actual capabilities
of such bugs in general in this respect.

345

It may be noted that whereas Kala azar has a very limited
distribution, the bed bug, in which early development has been
observed, is world wide in its range. If an insect transmitter of
Kala azar is to be found, it is probable that it will be more restricted
in its distribution than is Cimex. Their localized distribution, their
seasonal dependence on certain forms of plant food and their evident
adaptability, point to bugs of the Pyrrhocoridae and similar families
as objects of study. I believe my observations and experiments
indicate the necessity for an exhaustive investigation of all such
forms, not only in countries where Kala azar abounds but also in
countries in which Tropical Sore occurs.

REFERENCES

Battou, H. A. (1906). West Indian Bulletin, Vol. VII, p. 64.

Butter, E. A. (1923). A Biology of the British Hemiptera-Heteroptera. London.

DE CuarMmoy, D, v’EmMMerez (1921). Bull. Ent. Res., Vol. XII, p. 181.

Fantuam, H. B., and Porter, A. (1915). Proc. Camb. Philosoph. Soc., Vol. XVIII, Pt. 2, p. 30.
FRANCHINI, G. (1922). Bull. Soc. Path. Fxot., Vol. XV, p. 161.

LaronT, A. (1909). Compt. Rend. Soc. Biol., Vol. LVI, p. tott.

Lamsorn, W. A. (1914-15). Bull. Ent. Res., Vol. V., p. 197.

LaveraN, A., and Francuint, G. (1913). Compt. Rend. Acad. Sci., Vol. CLVI, p. 423.
Mawnsriecp-Apers, W. (1919-20). Bull. Ent. Res., Vol. X, p. 145.

Maxwett-Lerroy, H. (1909). Indian Insect Life, p. 665. London.

Patron, W. S., and Crace, F. W. (1913). A Text-Book of Medical Entomology. London.
Peacock, A. D. (1913-14). Bull. Ent. Res., Vol. IV, p. 192.

Ropertson, M. (1911-12). Proc. Roy. Soc., Ser. B, Vol. LXXXV, p. 234.

Rovpaup, E. ,and Francnini, G. (1922). Bull. Soc. Path. Exot., Vol. XV, p. 404.
STEPHENS, J. W. W. (1915). Transactions Liverpool Biol. Soc., Vol. XXIX, p. 3.

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347

TYPHUS FEVER IN GREEK REFUGEES

mMATOR-GENERAL..oIR, PATRICK. HEHIR,..K.C.1.E.,...C.B.,
Cie ool) Pole ele l’ My Lin,

MEDICAL ADVISER, BRITISH RED CROSS SOCIETY IN GREECE
(Received for publication 6 July, 1923)

During the last six months my duties in Greece have brought me
into contact with about one thousand two hundred cases of typhus
fever. From the nature of my work I saw the disease mainly from
the standpoints of the medical administrator and the sanitary officer
endeavouring to stamp it out in the areas allotted to the British
Red Cross Society.

Lpidemiology. Vhe Greek refugees primarily brought the
disease with them from Asia Minor and the Near East. With us
the epidemic began at the end of January, 1923, gradually rose in
number and severity of attacks, reached a maximum about the first
fortnight in March, remained stationary a month or so and is now
(June 22nd) dying out.

It is noteworthy that in the areas in which the refugees have been
widely dispersed on the land there has been no typhus, or it has not
gained a foothold when introduced. The brunt of the epidemic has
been borne by the larger and medium-sized towns, the smaller either
escaping, or being only slightly invaded. The city of Athens and
its environs, Salonica, Patras, Corfu, and many other large towns,
have suffered severely from the disease, whereas whole rural districts
in Macedonia and Western Thrace have not been affected at all.

The housing conditions of the refugees, the hardships they have
experienced, especially the insufficient food and exposure, have
considerably reduced their stamina and vitality, and lowered their
resistance to disease. |

The outbreak attained its maximum of intensity in the mid-
winter. In the colder weather the people are more crowded together,
they wear thicker clothes which foster lice, and they bathe less
frequently. Throughout the winter the refugees were packed with
the ordinary civil population in the waiting-rooms of railway stations

- 348

and in other places; contact causes were therefore more frequent.
In some small towns the number of refugees alone is equal to the
former population; in others the refugees out-number the permanent
inhabitants by two, or even three, to one. In the Landgada Valley
(near Salonica) overcrowding has reached its possible limits. The
ordinary population of the village of Landgada is five hundred;
during the last six months one thousand six hundred refugees have
been added to it. In one small refugee room, gft. by 13 ft. as
regards floor space and 5 ft. 6ins. to the roof, I found four adult
women and four children from 7 to 13 years old living; all the
husbands were prisoners with the Turks; this was one of many
similar overcrowded rcoms above cowsheds. One could multiply
instances of this sort; but overcrowding is only one of the many
economic factors that enter into the epidemiology of typhus.

What has been happening is that infected refugees are necessarily
brought into intimate contact with their non-infected comrades, all
of whom were at one time (and to some extent are still) lice ridden.
Infected lice are transferred to uninfected refugees. In the over-
crowding that has arisen in nearly all the larger towns of Southern
Greece, lousy infected refugees have also been brought into contact
with the people of the permanent community, and the disease has
thus spread to them. On several occasions I have seen, in organised
refugee camps, typhus fever cases living with the uninfected and
sharing the common bed, which is practically always the floor of
the room or block they inhabit. The same takes place in railway
stations where verminous typhus refugees infect the ordinary
passengers in the waiting-rooms. Other hardships exist. A very
large percentage of the refugees get only a bare subsistance allowance
of food. There are districts in which the men who can work get no
food dole and the out-of-work women only about six to eight ounces
of bread or flour a day. For some months few refugees had a
change of clothing; undergarments were especially scarce. With
only one suit of clothes the practical difficulties of delousing are
obvious; it is put through the steam disinfector and dried while
the refugee is having his bath. It is necessary to emphasise that
the sanitary condition under which the refugees lived for months
after their coming to Greece was very bad; it has been greatly

ameliorated.

349

Mode of Lransmission. The transmission of typhus by body and
clothes lice may be accepted as proved; the infected human louse is
the intermediary in the transmission of typhus from typhus cases to
the healthy—ordinarily there is no direct communication of the
disease from man to man. If it were possible to exterminate human
lice in an infected area, the disease would cease. Further, if we
could rid all typhus cases of lice the disease would come to an end.
All our radical preventive measures are based on these facts.
Somehow it has got abroad that it is only the body-louse that acts
as a carrier. This is not the case; the exculpation of P. cafitis is
a dangerous and pernicious theory to inculcate.

Period of Incubation. A generation or so ago, twelve days used
to be given as the period of incubation. All recent experience of
the disease in Russia and Greece shows this to be correct. It has
been proved by the inoculation of human blood experimentally ;
inoculation of monkeys with infective human blood has likewise
demonstrated it.

Most cases are admitted into infectious diseases hospitals on the
fifth day of the disease, the next most frequent is the sixth day.
- This late admission arises from several causes—antipathy of the
refugees to typhus hospitals, their ignorance as regards the nature
of the disease, inability to obtain medical advice, overwork of the
doctors, etc. In all but a very small proportion of cases the eruption
is out by the time the patients reach the hospital.

Symptoms. In the vast majority of cases there are prodromata
in the form of ill-defined malaise with vague symptoms for two or
three or even four days before more severe indications arise. The real
onset is well-defined. In typical cases the patient knows the day,
often the hour, when he first felt genuinely ill and had to go to bed.
The face is then somewhat flushed, the conjunctiva injected, the
expression excited or dull, the tongue is coated, the lips and mouth
dryer than normal; thirst, constipation, severe headache, pain
in the back and limbs are complained of. Constipation is present
in the great majority of cases, and often persists throughout the
illness. By the time the patient is brought to hospital (fifth or sixth
day) there is as a rule no doubt about the diagnosis. By the fifth
day the mucous membranes are often implicated in the rash. The
tongue has a well-marked white coat and is tending to become dry;

35°

later the tongue is fissured, the mouth becomes offensive, sordes
collect on the teeth, although these latter conditions can in many
cases be prevented by proper nursing. Diarrhoea is not common in
the early stage; about 20 per cent. of the cases develop it in the later
stages. Asthenia and muscular debility are always present ; patients
can scarcely move in bed, they are often unable to protrude the
tongue, and lose all expression. Emaciation in some cases towards
the end of the disease is marked. In a small proportion of cases
there is a definite crisis; 1n most cases, however, there is lysis,
but a mixture of these beginning with lysis and ending in a crisis
or the reverse may take place. The most common symptom of the
real onset 1s headache, usually frontal, but sometimes mainly
occipital. Conjunctival injection is present in four-fifths of the
cases; 1t increases with the development of the eruption until lysis
begins. Vomiting 1s present in about 25 per cent. of the cases.
Sometimes it 1s severe and persisting for several days.

In many cases there is a distinctly reddish blush along the edge
of the soft palate and pillars of the fauces, less frequently also slight
congestion of the throat.

The Temperature. \Nhilst 1 believe it 1s possible to construct
and describe what might be considered a normal temperature chart
for an average case of moderately severe typhus, it is seldom that
such a chart is met with in the wards in the natural course of the
disease. A continuous or slightly remittent temperature of 103° or
103'5° F., during the first half of the second week, and then a slight
daily decline until the eleventh or twelfth day, when there is a more
decided remission with abatement of all the symptoms, is ordinarily
what may be expected. Then there is another rise, say on the
twelfth day, and a remission, and a second similar oscillation
though less marked, with a decline to normal, and even a third, the
whole lysis occupying forty-eight to sixty or seventy-two hours.
Irregular temperatures are also met with. Again a definite crisis
with a fall of temperature to normal in twenty-four or thirty-six hours
may occur, although this is exceptional. After the temperature has
dropped and the symptoms have disappeared, the drop may be to
sub-normal for some days.

Of two hundred and forty-six cases that recovered, in nineteen
the temperature was normal on the twelfth day, thirty-seven on the

351

thirteenth day, sixty-six on the fourteenth day, forty on the fifteenth
day, and twenty-two on the sixteenth day.

Lhe Pulse. Normally in ordinary cases, the pulse-curve follows
that of the temperature. The pulse, however, is liable to show much
variation; sometimes marked oscillations occur in the twenty-four
hours, being at one time ninety and at another one hundred and
twenty to one hundred and thirty in a minute. Dicrotism is not
uncommon, especially towards the end of the second week. With
the tendency to cyanosis so commonly seen in the late stage of the
second week, the pulse is often absent at the wrist. The state of the
patient’s lungs appear to me greatly to affect the pulse, especially in
wide-spread broncho-pneumonia. The pulse is markedly improved
on the first signs of defervescence; in a few a very slow pulse is
present in convalescence. The respiration curve varies less than that
of the pulse.

Lhe Respirations. In uncomplicated cases with moderate
temperatures, the respirations are shallow and from thirty to thirty-
five per minute, they vary little and without real dyspnoea at any
stage. In similar cases with broncho-pneumonia, dyspnoea becomes
a serious symptom.

The Eruption. ‘This first appears on the evening of the fourth
day in the form of discrete and well-defined pink or roseolar spots
which may be round, oval or irregular, varying from 2 to 5 mm. in
diameter, vanishing on pressure; they are seldom palpable at this
stage, but they are widespread though scanty, and are seen on the
abdomen, back, chest, shoulders, arms, legs and feet; they are rare
on the face and head. In this early stage the rash described 1s
not very obvious, it may require careful scrutiny to find it. The
macules then become larger and of a bright red colour, next assuming
a purplish-red hue running into dark purple. At this stage the
tendency is for the eruption not to disappear on pressure, but this
is not invariable—in many cases ending fatally with a deep coloured
eruption before death, no sign of it remains post-mortem. When
the eruption is fully developed on the eighth or ninth day, well-
defined dark coloured patechial areas which do not disappear on
pressure are seen, besides less-defined patches of much lighter colour
which do not disappear on pressure. In all severe cases with typical
eruption, these erythematous and patechial patches are met with

352

during the second week of the disease. In blonde boys and girls,
during the early stage of the disease, we sometimes see on the chest,
neck, arms, and occasionally on the abdomen, an irregular or blotchy
erythema which vanishes before the real eruption is developed. In
the second week the eruption has a multiform character—roseolar
patches, red spots, maculae, small patechiae and large plaques
typically patechial are seen; this multiformity is well seen on the
shoulders and back, lower part of the abdomen and hips, outer
surface of the arms and forearms, on which places what has been
admirably named ‘subcuticular mottling’ is also visible. The
eruption may, however, vary from consisting of only faint roseolar
slightly raised spots to large ecchymotic looking patches. By the
end of the second week little of the eruption is left. In some cases
the general lousiness antecedent to the onset of the disease leads to
considerable skin irritation with scratching and local secondary
infections, which may initially be rather puzzling. Chronic
pediculosis and pityriasis versicolor (both common in refugees) are
the chief conditions of the skin hkely to lead to confusion in a
diagnosis based on the eruption alone. In about 1 per cent. of
typhus cases there is either no eruption or only a faint roseolar one;
this is more frequently the case in children and adolescents ; in these
cases the Wiel-felix reaction is present. It is useful to carry about
a good hand-lens, and, to bring out the eruption, rub into the skin
some petrolatum ; the lesions are then seen to consist of a congeries
of dark red blood vessels.

Insomnia is one of the commonest symptoms; the majority of
cases suffer from it during the first week of the disease.

In about 25 per cent. of the cases some form of mental disturbance
is present on admission, and on the seventh or eighth day delirium.
In cases that are running a fatal course, the delirium often passes
into coma more or less complete. Cough is one of the most constant
symptoms. In the earlier stage it is short and dry. Later on the
expectoration may become profuse and mucopurulent. In a number
of cases patches of lobular pneumonia occur. This is a common
terminal condition in fatal cases. Diarrhoea is common in the later
stages of the disease, and is then sometimes associated with rectal
incontinence. Parotitis 1s one of the more serious complications;
I saw altogether twenty of these cases, and as many as three in a

353
ward of thirty patients. Otitis media occurs in a small percentagé
of cases; it may become chronic. Deafness is a marked feature in
many cases of typhus, during the late stage of the disease; this is
quite distinct from the dullness of intellect that exists during that
stage.

The spleen can be felt in about two-fifths of the cases ; sometimes
it is of considerable size. I do not lay stress on this, as many
patients have a history of old malarial infection.

In all hospitals dozens of recurrent fever were sent in as typhus.
In relapsing fever the sudden onset with rapid rise of temperature,
severe headache, pains in the back and extremities, absence of
dulness and apathy and (ordinarily) of rash, the presence of a moist
tongue and of S. obermeierz in the blood, and later in the disease,
more or less anaemia, should be suffhciently distinctive. In the
Salonica Hospital the records show that eight cases of typhus and
relapsing fever ran their course concurrently in the same persons.

The use of neo-salvarsan for the recurrent fever did-not affect the
normal progress of the typhus. The combined infection seems to
suggest that the same louse may be able to carry the virus of typhus
and S. obermeieri and inoculate them at the same time. Of course,
two or more lice, each with a single infection, may have attacked
these cases. It seems to be established that, contrary to the rule
with intermediate hosts, the virus of typhus eventually kills the
louse. In many, typhus and influenza ran together, the latter
disease being also epidemic at the time. Hundreds of cases of both
smallpox and typhus have been admitted into infectious diseases
hospitals ; in no single instance have both diseases been met with in
the same patient at the same time; in one, typhus followed smallpox
from infection acquired in the hospital.

Wiel-Felix Reaction. In typhus this reaction is very distinctive.
It owes its origin to the discovery of the fact that what are called
the ‘X’ strains of B. proteus are agglutinated by the serum of
typhus cases. The special strains that do this are X2 and X19;
these were priinarily obtained from typhus urine. The macroscopic
method is here usually adopted. The minimum dilution accepted
as positive is I to 100. Practically the serum of every typhus fever
case after the eighth day is positive, whilst that of other fevers 1s
negative.

354

Complications. Bronchitis of greater or lesser severity is
present in a large proportion of cases. Broncho-pneumonia is
another common complication affecting the bases of both lungs.
Pleurisy is much less frequent. Other complications are—severe
diarrhoea, myocarditis, cardiac dilatation, parotitis, otitis media,
conjunctivitis, keratitis, gangrene of the toes, bedsores, etc.

Prophylaxis. Medical men, nurses, and all sick attendants
looking after typhus cases should be thoroughly protected from lice
by suitable white cotton or linen clothing from head to foot before
commencing their work, ard have a bath and complete change
of clothing after finishing their day’s work. Even with these
precautions infections will occur, but without them infection is all
but certain, sooner or later, in those not immunised artificially, or
by a previous attack of the disease.

Etiology. It would appear that the micro-organism of typhus
passes through a development stage in the louse; in that insect it is
intracellular, develops, is set free and is introduced into man. In
man it is said to become intracellular once more, and from the
infected cell to be thrown into the blood with its toxins. The
members of the Typhus Fever Commission of the League of the
Red Cross Societies to Poland, however, found no evidence of the
development of the micro-organism in the louse, but they arrived at
certain important conclusions as the result of their work.

Summarised, their conclusions are :—

Pathology. The lesions of typhus appear to be situated in the
blood vessels of the skin, central nervous system, skeletal muscles,
and to a lesser extent in some of the viscera—heart, kidney and
testes. Typhus is considered to be a disease of the smaller blood
vessels, and localises almost exclusively in the vascular endothelium.
The reaction to the parasite is shown primarily by degenerative
changes giving rise to thrombi in the blood-vessels, and by a
proliferative reaction on the part of the endothelium and neuroglia
which give rise to the characteristic ‘nodules’ of the disease in the
skin and central nervous system. When lice are fed on typhus
cases, while they develop Rickettsia prawazeki with great regularity
they develop no other form of micro-organism. All lice so far do
not become infective, but why this is so is not determined. Infection
with R. prawazeki eventually kills the louse, which is an exceptional

355

effect of a parasite upon its intermediate host. R. prawazeki
escapes from the alimentary tract with the faeces, and therefore may
be introduced by scratching or by the mouth-parts of the louse
becoming soiled with the faeces. Rickettstza has not been found in
the salivary glands or in the mouth-parts of the louse.

Mortality. The average mortality in the infectious diseases
hospitals near Athens 1s roughly 10 per cent. ; it is, however, higher
in some towns, such as Corfu, Patras, Volo, etc. It varies also
greatly at different ages. In refugees of 50 years and over, the
death rate is high, reaching in some towns 50 per cent.

It is necessary that some definite routine plan should be adopted
in admitting typhus fever cases into hospital and distributing them
in wards. The first requirement is a receiving-room, to which all
patients are primarily brought. Here the hair of the head is rapidly
cut off with a machine clipper, the hair of the axillae and pubes
being shaved cff; the hair is to be burnt. The receiving-room
should communicate with the room or other area containing the
steam-disinfector on the one hand, and with the bathroom on the
other; this latter should lead to the dressing-room. After removal
of the hair the patient is put on a stretcher and conveyed to the
room containing the disinfector. Here he discards everything that
he brings with him, which is disinfected. He is taken to the
bathroom and bathed. He is then put on a clean stretcher and
removed to the dressing-room; here he receives a suit of clean
hospital clothing and is taken to the ward he is to occupy. There
must be no remission in this routine; it must be thoroughly carried
out if the wards are to be kept free from infected lice. Nothing
that the patient brings with him to hospital should enter the ward.

Thoroughly deloused typhus cases are perfectly innocuous to the
uninfected, and if we are quite confident as regards the efficiency
of our delousing arrangements there is no reason for putting them
in different wards.

Treatment. Typhus patients should be kept in bed throughout
the pyrexial stage of the disease, and for a fortnight after the fever
has subsided. Constipation is best relieved by a simple enema every
second day. In some cases the catheter has to be used to drain off
the urine. In the early stage, when sleeplessness, irritability,
general discomfort and delirium are present, small doses of morphine

356

give satisfactory results. The morphine may later on be replaced
by veronal, sulphonal, paraldehyde or chloral hydrate, if one or
other is called for in mild delirium or insomnia. For prolonged
and marked active delirium, hyoscine, hypodermically, is a valuable
drug. The most popular stimulant is hypodermic injections of
camphor (5 grains in I c.c. of olive oil or ether put up in ampoules) ;
strychnine is also largely employed.

Prophylactic Inoculations. As a substitute for a prophylactic
vaccine the blood of typhus fever cases has been inoculated, and the
virus thus introduced in a living state. In using the living virus,
Kusama injected monkeys with a fairly definite minimum dose of
typhus blood, known as the minimal morbid dose, to bring about
an attack. Ifa smaller dose is given no attack occurs, but a state
of immunity is produced and the animal can tolerate many times
the minimal morbid dose without ill-effects. This active form has
so far not been used; the killed virus is the one that is used here.
Whether it has any prophylactic value 1s uncertain.

Prevention. ‘The amount of actual physical labour connected
with the preventive work associated with typhus may be understood
by describing what took place in two large blocks housing nearly
two thousand refugees, in March last. From the 12th to the 17th,
the whole camp was deloused and all bedding and clothing put
through the steam disinfectors. The entire floors were washed with
disinfectants and the walls whitewashed. This meant that every
room had to be emptied of its entire contents while this was going
on; then all the belongings of the refugees returned to the cleaned
rooms, which were shut off from those awaiting their turn. It
likewise meant giving a complete hot bath to everyone in the camp
under supervision. From the 1oth to the 24th there was a renewal
of the bathing. A slight recrudescence of typhus after this
necessitated a repetition of the processes carried out previously.
This was done from the 26th to the 31st March. All men’s and
boys’ heads were shaved, and all girls up to the age of fourteen had
their hair bobbed. All cases of actual typhus, of course, had their
heads shaved. Our nursing sisters also used on the heads of the
refugees in camp the mixture of equal parts of kerosene and olive oil
we employ to free nits from hair, and they distributed N.C.1 powder
(naphthaline ninety-six parts, creosote two parts, iodoform two parts)

357

which was in little bags to be worn for a week, when a fresh bag is
issued. The idea is that the heat of the body helps to vaporize the
ingredients, and in this way 1s created a louse-destroying atmosphere
next the skin and the clothes.

By order of the Government, refugees are allowed to leave the
camps to work or to search for work, so long as their identity cards
show that they are free from infectivity, which means free from lice.
Many of the people allowed outside run the risk of acquiring the
disease in other refugee camps which they visit, or elsewhere. This at
present is unavoidable. Another weak link in the chain of preventive
measures was the fact that for months after their arrival in Greece,
a large percentage of the refugees did not possess a change of
underclothes. It is obvious that where the people have only one
suit of clothing there must be grave difficulties in rendering them
lice free. An adequate supply of steam disinfectors for delousing
clothes and bedding is almost indispensable; in their absence the
task is most laborious. In small communities, Serbian barrels and
other such improvisations may be useful, but in dealing with large
masses of people they are futile.

I believe it to be well worth while in every camp to endeavour
to educate the people in regard to the nature of typhus and
the principles that underlie its eradication and prevention. We
placarded leaflets in this connection, and also had them read out
periodically for the benefit of the illiterate.

The specific measures now indicated appear to be to disperse the
refugees from overcrowded towns to rural areas as much as possible.
Another urgent requirement in many towns 1s an increased supply
of water, which should be available to every house, or at least be
_ within easy reach. In some camps the water-supply is decidedly
defective, which interferes with the bathing arrangements, steam-
disinfection of clothing and bedding and ordinary laundry-work—
serious obstacles when endeavouring to eradicate typhus fever from
an infected camp. The Greek Administration is endeavouring in
all possible ways to remove this difficulty.

Every effort to quarantine contacts has failed, and all things
considered it is not reasonable to expect it. All we want is that
the person who is a contact should be watched definitely for
twenty-one days, that is the time we laid down as the incubating

358

period. We endeavoured, as far as was possible, to render these
certificated people free from lice; we felt that if thus free they could
not communicate typhus, even if they acquired it themselves in the
meantime. The quarantining of contacts in a small country which
contains over a million refugees, many thousands of whom are
contacts and are either in search of work or have daily to go to
work from their camps and return to them, is an impossibility. It
may be easy to do this in a limited area of infection with a stationary
and disciplined population; it is an impossibility among refugees.
The hope is that nature will step in during the summer and bring
the epidemic to an end by stopping the multiplication of lice, by
unfavourable meteorological conditions, and that by the beginning
of the next typhus season the economic and other conditions will
have altered for the better. At the present moment it 1s certain that
the economic state of the refugees is decidedly antagonistic to
effecting a cessation of the disease by the adoption of ordinary
preventive measures.

REFERENCES

Kusama, S. (1921). Lancet. Vol. CCI, p. 886.

Wotzacu, S. B., Topp, J. L., and Parrrey, F. W. (1922). Report of the Typhus Fever
Commission of the League of Red Cross Societics to Poland. Harvard Univ. Press,
Mass.

359

mot VALUR OP THE’ SACHS-GEORGI
REACTION IN THE SEROLOGICAL
DIAGNOSIS OF SYPHILIS

GEORGE STUART sMaA NeBAY Chi By

(ASSISTANT DIRECTOR OF HEALTH, GOVERNMENT OF PALESTINE.)
(Received for publication 6 July, 1923)

In order to determine to what extent the Sachs-Georgi could
be relied upon to replace the Wassermann reaction in this country
(Palestine), | commenced to perform a parallel series of tests in the
latter part of 1921. It was by no means difficult to secure suitable
material, inasmuch as it was possible to collect the sera of a large
number of untreated cases which gave true clinical manifestations ~
of syphilis 1n one or other of its various stages and forms. It was
not so easy, however, to gather together sera from cases undergoing
treatment, nor to follow up the effect of such treatment on the
reactions.

Doubtless the disappearance of the then existing lesions led
those affected into the belief that they were cured, and this deduction
of ‘ out of sight, out of mind’ is strengthened by the fact that only
comparatively few names appear on the register more than once.

The conclusions arrived at, both from the performance of the
ordinary tests and from certain experiments carried out bearing on
the subject, are of interest, if, in one or two particulars, somewhat
puzzling.

I shall in this paper detail :—

(a) the methods employed—aincluding the technique of the

Sachs-Georgi reaction as carried out during the whole series ;

(which technique I have found by experience easiest and most
suitable) ;

(4) the actual results of the two reactions ;
(c) the relative percentage of agreement ;

(d) certain fallacies of the Sachs-Georgi reaction—certain
experiments made with a view to explain the cause of several,
and certain theoretical observations; and finally,

(é) the conclusions and inferences drawn,

360

(A) METHODS EMPLOYED

For the performance of the Wassermann reaction, the Boas
modification of the original has always been employed; the results
of this technique have been so uniformly dependable that no other
method is permitted in these laboratories. The antigen and
haemolytic serum are both prepared by Burroughs, Wellcome & Co.,
and are forwarded every three months.

The technique of the Sachs-Georgi reaction is simplicity itself,
and will be briefly outlined here.

(a) Antigen. This is supplied quarterly by Burroughs,
Wellcome & Co., and although, in 1921, I experimented with
various antigens, I found that prepared by the above firm so reliable
that it was adopted in preference to the others.

A dilution of 1 in 20 is required.

I c.c, antigen is pipetted into the ordinary 1 inch by 6 inches
test-tube. To this is added 1 c.c. normal saline freshly prepared—
the saline being allowed to run slowly down the side of the test-tube
held in the sloping position.

The tube is agitated gently during the process of admixture.
18 c.c. normal saline are now to be added. The test-tube is held
vertical and a pipette containing the 18 c.c. held with its point
midway over the upper end of the tube. By gentle pressure on the
indiarubber teat, the saline is allowed to fall drop by drop the
height of the test-tube into the mixture; during the whole time this
is being effected, the tube is gently shaken from side to side. The
tube now containing 20 c.c. of 1 in 20 dilution is inverted slowly
against the palm of the hand several times. The resultant antigen,
ready for use, is a shimmering, somewhat opaque fluid with just a
milky tint, giving the appearance of watered silk.

(0) Patient’s serum. The serum is inactivated as in the
Wassermann reaction, for half an hour at 56°C. When sera have
to be sent to the laboratory from a distance, the medical officers are
issued with instructions that blood must be taken with all aseptic
precautions, and that the serum should be allowed to separate out
in a sterile test-tube (preferably kept over night in the sloping
position) and transferred to sterile bottles which are then carefully
stoppered and sealed. In this way not only is haemolysis prevented

361

but also 1s obviated one of the chief causes of failure (to be detailed
below) of the Sachs-Georgi reaction and discrepancy between this
reaction and the Wassermann.

(c) Method of setting up the reaction. Four Wassermann tubes
are used for each case, and dilutions of patient’s serum made,
I in 5, I in 10, I in 20, and 1 in 4o, with normal saline freshly
prepared.

Into tube 1 is placed 1°6 c.c. and into tubes 2, 3, and 4, I c.c.
of normal saline.

To tube 1 is added 0°4 c.c. patient’s serum and the contents of
the tube are thoroughly mixed with a pipette. One-half the contents
of tube 1 is transferred to tube 2 and thorough mixture made again.

To tube 3, one-half the contents of tube 2 is added, and again,
after mixture, one-half the contents of tube 3 is transferred to
tube 4. After suitable mixing, one-half of the tube 4 contents is
now discarded.

Tube 1 now contains 1 c.c. of 1 in’ 5 dilution of patient’s serum.

mube 2 ,,; Pe A KeKon Mein Lethe ee) a 5g 3
ube 3°,; oe LICyCat Ofel 1ne20 :% 55 ie
Buber 45%), BA I “cle. fofe1 1n;40 4 5 h

To tubes 1, 2, 3, and 4 is added 0’5 c.c. of the already prepared
1 in 20 dilution of antigen, and the contents of all tubes are
thoroughly mixed by inverting the tubes between the thumb and
different fingers several times.

The tubes are now placed in an ordinary Wassermann bath and
kept at a temperature of 37°C.

Mixing by inverting the tubes is done as a routine measure
three times during the first twenty minutes, unless signs of a positive
reaction have already made themselves manifest.

At the end of one hour, six hours, eighteen hours and twenty-four
hours, the results are read and recorded. Saline negative and
positive controls are set up in similar fashion according to the
method described above. 1 c.c. of normal saline replaces the I c.c.
patient’s serum in the saline control, while 04 c.c. of known
negative and positive sera will be added respectively to the 1°6 c.c.
normal saline in the first tubes of the series instead of 0°4 c.c. of
patient’s serum.

For the positive control is taken one-half of a serum previously

362

proved to be positive both by the Wassermann and Sachs-Georgi
tests.

READING OF RESULTS.

In well marked cases there is no difficulty, even for the most
inexperienced, in reading both positive and negative reactions.
The negative tubes show at the end of eighteen to twenty-four hours
the same uniform shimmering fluid which on agitation presents the
appearance of watered silk. The impression of the term ‘ watered
sik’ must be very carefully appreciated by the beginner, as upon
that impression depends future success or failure in reading results.

The use of a hand-lens in doubtful cases is advocated and has
proved of service to my assistants, but personally I have found the
readings of ‘ granular positives’ relatively simple, as my own visual
acuity is — 5D.

Positive results, if very marked, give the appearance of snow-
flakes suspended throughout a clear fluid, and from the gradual
settling of these white masses, a snowy layer ultimately forms at the
bottom of the test-tube.

The supernatent fluid is then absolutely clear. The flocculations
in this class are termed massive, and the precipitate heavy; we
register it as XXX.

If defintely positive but less marked, the tubes give the
appearance met with in ordinary bacterial agglutinations when a
high titre serum is used.

The flocculi tend to deposit later and leave a clear supernatent
fluid; this we regard as XX.

In the third group considerable difficulty may be experienced in
differentiating the finely granular flocculi of a positive from the
homogeneous suspension of the antigen. The appearance presented
is somewhat like particles of dust shewn up by a ray of bright
sunlight suddenly penetrating a dark room. There is little or no
tendency for these particles to deposit, and the fluid does not become
clear as in positives described above, this we record as X.

If doubt still exists, recourse may be made to having the tubes
centrifuged to see whether a deposit can be obtained.

It has not been considered necessary to adopt any artificial
lighting device to help in the reading of the results.

In passing, it may be remarked that on numerous occasions tubes

363

have shown what appeared to be positive reactions within the first
few hours, but at the end of eighteen to twenty-four hours this
appearance had completely disappeared; and also, it has been noted
that gentle agitation may produce such a disappearance in what
may be called pseudo-positives.

I shall have occasion later to refer to the occurrence of positive
readings in the Sachs-Georgi reaction—actual positives which do
not disappear as those mentioned in the last paragraph, but which.
owe their existence, although not to syphilis, to certain definite
causes.

(B) ACTUAL RESULTS

1. Complete POSITIVE agreement was obtained between the
Wassermann and Sachs-Georgi reactions :—
(a) In three hundred and ten uxztreated cases submitted from
venereal clinics and hospitals with a definite history of syphilis.
Analysis of these three hundred and ten cases :—
(1) Primary stage. Forty cases.
(2) Secondary stage. Two hundred and fifty cases.

These cases without exception showed typical pictures of
secondary syphilis-rash, sore throat, mucous patches, etc.
They had neglected the primary stage completely, and had
come to seek advice only when the rash, fever, and constitu-
tional symptoms manifested themselves.

(3) Lertiary stage. Eight cases.

(4) Congenital syphilitics. Five cases.
These were discovered during the routine examination of
inmates of an orphanage.

364

(5) In seven cases where the patients had several abortions.

Taste IT.
Wassermann Sachs-Georgi
Case History reaction reaction
1 | 3 abortions; 2 still births... ae ni =f ee XXX XXX
26 aportions a... fee 4 ms pie Bs sta XxX XX
3 | 6 abortions ... ay e i%j oe ra ap XXX xX
4 | 5 abortions ... aa a se ie ets 3% XXX XX
5 | 2abortions ... “i ae By ee yy ne XXX XXX
6 | ‘Several abortions’ ae ae re es ms XXX XX
7 | Several abortions’ es fee a Vs - XXX xX

In case No. 1, husband had syphilis five years ago, and
was treated with full course of Neo-Salvarsan injections.
Serum tested by both reactions on same day as patient gave
completely negative readings.

(2) In certain ¢veated cases.

Eight cases showed markedly positive reactions. Of these cases
one had received two injections of Neo-Salvarsan, and four had
had ‘complete’ treatment (see below).

2. Negative agreements.

Negative readings in all tubes of both reactions were obtained in
five hundred and fifty cases of sera submitted without definite
history, and as part of routine examinations. In two cases
previously reacting XXX to both tests, sera now are completely
negative to both.

3. Partial agreements.
These may he best illustrated as follows :—
Taste III.

Wassermann Sachs-Georgi
Cases reaction reaction Remarks

4 XXX X ‘I with 4 injections of Neo-Salvarsan.

305
4. Non-agreement.

(a) In forty-five cases submitted, mostly without history, the
Wassermann reaction was definitely positive, the Sachs-Georgi
negative. In thirteen cases, however, details were supplied-
symptoms of primary stage 8, of secondary stage 5.

(4) In twenty-one cases the Sachs-Georgi reaction was positive,
the Wassermann completely negative. This figure does not
include positive Sachs-Georgi reactions obtained during experiments
performed. These latter will be detailed below.

(These cases were either partially or completely treated according
to the following routine method practised here.

The course of Neo-Salvarsan injections is :—

Ist injection ... O45 grammes.
2nd injection ... 0°60 grammes one week later.
3rd injection aos ie .* % a
Ath injection ee 60,90 i 3 ie <
5th injection ol 10-00 re ¥ o eae hs)
Tasre LV.
No. of cases Wassermann reaction Sachs-Georgi reaction Remarks

14 XX fe) ‘ partially ’ treated

Z XXX fe) ‘with 3 injections’

7 xX | fo) | « complete treatment’

(C) THE RELATIVE PERCENTAGE OF AGREEMENT BETWEEN
THE REACTIONS
The calculations are based upon one thousand and thirty-seven
examinations of sera.
1. Positive agreement.
In all, four hundred and eighty-seven showed a well-marked
positive Wassermann, while with the corresponding sera the Sachs-

366

Georgi reaction showed positive readings in four hundred and
nineteen.
The positive agreement therefore is 86 per cent.

2. Negative agreement.

Whereas the Wassermann reaction was negative in five hundred
and fifty cases, the Sachs-Georgi was negative in six hundred and
eighteen.

The negative agreement is therefore 89 per cent.

(D) CERTAIN FALLACIES OF THE SACHS-GEORGI REACTION

WITH CERTAIN EXPERIMENTS MADE WITH A VIEW TO EXPLAIN

THE CAUSE OF SEVERAL, AND WITH CERTAIN THEORETICAL
OBSERVATIONS

(1) The presence in the serum to be tested of contaminating
organisms renders the findings of the Sachs-Georgi reaction of no
value whatever.

The first disparities between the Wassermann and Sachs-Georgi
reactions here occurred in cases where the sera were sent from a
distance, and when two or three days elapsed between collection
and examunation.

In those cases, fortunately, little doubt could exist as to
contamination, as the sera were turbid and malodorous on arrival.
Actual proof was obtained by plating on culture medium. A new
series of reactions was performed in the case of those sera proved
contaminated: they were re-submitted, and after transmission,
arrived in a sterile condition. On these occasions they showed a
completely negative reading where previously they had shown a
strongly positive reaction. I refer in this connection only to sera —
which reacted positively to the Sachs-Georgi when the Wassermann
reaction remained negative. |

Further proof was adduced by the following simple experi-
Tico

A normal blood was drawn off in the laboratory, and the serum
proved negative by both reactions.

The serum was then artificially infected with (a) B. typhosus
and (4) B. subtilis, and then after suitable incubation the sera were
set up in the ordinary dilutions, with the following results :—

Sc ~

367

TABLE V.

Wassermann | Sachs-Georgi
Case reaction reaction Remarks

I Normal blood serum be ° °

2 | The same serum infected

WHER Be SUDEES on doc 9 xe ° XXX fa both cases flocculation
massive, precipitation
3 | The same serum infected { heavy.
with B. typbosus ... aes ° XXX

If, then, the reaction could be completely altered by the presence
of contaminating organisms in the serum, I considered it advisable
to test the sera of patients suffering from certain diseases, to
determine whether positive results in the Sachs-Georgi might be
obtained here also. |

The results are of interest, and although unfortunately at
present the number of such examinations is small, yet I hope later
to submit the results of examinations of many sera collected from
patients suffering from the diseases most common in Palestine, so
as to determine to what extent the Sachs-Georgi reaction is
influenced by such.

First I was able to obtain six sera from lepers in Jerusalem, and

. the particulars and reactions are as follows :—

Tasce VI.
| Wassermann | Sachs-Georgi
Case Treatment | reaction reaction

1 | Nodular leprosy ...) Injections with Ol. Chaulmoograe

for 1% years as ay, a O° °
2 | Nodular leprosy ...| Injections with Ol. Chaulmoograe

From these examinations, no conclusions can be made.

368

In addition, however, the following examinations were made all
at the same time with the fullest possible controls. All precautions
were taken to ensure (a) freedom of the sera from contamination,
except where otherwise indicated; (4) that the reactions were made
in the dilutions considered essential; (¢) that the reaction of the
normal saline used was PH7; (d) that the readings were made after
one hour, six, eighteen, and twenty-four hours, and that pseudo-
flocculation was eliminated.

As these examinations have been performed for experimental
purposes, the results have not been included in the calculations
made as regards positive and negative agreements.

Tasre VII.
No. of Wassermann | Sachs-Georgi
case History reaction reaction Remarks

1 | Street case taken at random Tae fe) °

2 | Negative control ... a) HE fo) re)

3 | Positive control... oes <Bs XXX XXX

4 | Hospital routine examination... ° °

s | Routine examination from clinic ... ° °

6 | Case of untreated syphilis (stage 2) XXX XXX

7 | Sore throat, rash, typical 2nd stage XXX fe) Disparity 1
8 | Aneurysm of Aorta a sie XXX ° Disparity 2

g | Case of syphilis previously reacting
strongly to both reactions, treated
with 4 injections of Neo-Salvarsan XXX XXX Treated according
to table above.

10 | Case of syphilis previously proved
positive to both reactions—treated

with 4 Neo-Salvarsan injections XXX XXX
11 | Sore throat, mucuous patches... XXX XXX
12 | Soldier—contracted eyphihs aise
fully treated ee an fe) fe)
13 | Soldier—Syphilis ues course
of. treatment ae ' fo) fe)
14. | Soldier—primary case chancre ... XXX XXX
15 | Soldier—ulceration of palate vs! XXX XXX

——

I A »

32
33
34

35
36

Taste VII—Continued.

369

History

Male—previous syphilis untreated

Female—married case 16 last year ;
has had a child full term, showing
signs of congenital syphilis

Untreated secondary stage syphilis

Arthritis (right ankle)

Syphilis—after full treatment

Periostitis of femur

Ulceration of buttock

Routine examination—no history

Pharyngitis ...

Chronic Jaundice

Tumour of R. hypochondrium
independent of liver; inguinal
glands enlarged ; pigmentation

of legs and ankles

General malaise
Pustular eruption

Enlarged spleen (malarial) ...

No history ..

No history ...

General malaise

Serum from case of Typhus fever
showing all clinical symptoms
and reacting to Weil-Felix test

(B. proteus X 19) 1 in 400

Serum from case of Typhus fever
Weil-Felix reaction 1 in 400

Serum from case of Typhus fever
Weil-Felix reaction 1 in 200

Wassermann
reaction

XXX

Sachs-Georgi
reaction

Remarks

XXX

Pseudo-flocculation
in first tube only.

Disparity 3

Pseudo-flocculation
in I tube only.

Pseudo-flocculation
in 1 tube only.

Disparity 4
Disparity 5

Disparity 6

3/2

Taste ViII—Continued.

No. of Wassermann | Sachs-Georgi
case History reaction reaction Remarks

39 | Serum from case of Typhus fever

Weil-Felix reaction I in 400... XxX Oo Disparity 7
|
40 | Serum from case of Relapsing fever fo) ie
41 | Normal blood serum ef =. fe) fe)

42 | Serum of 41 infected with
B. typhosus ace <i YE ° XX Disparity 8

43 | Serum infected with B. subtilis ... ° XX Disparity 9

44 | Serum of patient suffering from
acute lobar pneumonia ... aes fe) XX Disparity ro

45 Serum—another patient recovering
from lobar pneumonia ... rf fe) XX Disparity 11

In the case marked pseudo-flocculation in tube 1, it was
found, as previously mentioned under ‘reading of results,’ that
gentle shaking of the tubes in question produced an immediate
disappearance of the seeming floccull.

The importance of the two precautions—(a@) always to shake the
tubes gently before reading; (4) not to give definite reading until
after the tubes have been set up for eighteen to twenty-four hours—
cannot be too strongly emphasized.

I do not attempt to give any explanation of the disparities
between the two reactions in the cases instanced above, but these
few findings would seem to suggest that the presence of organisms,
or products of organisms, in the patient’s blood might well have
some effect in reducing the value of the Sachs-Georgi unless the
fullest history accompanies each serum submitted.

(2) Granted the complete sterility of the sera submitted, could
any reason be advanced for the Sachs-Georgi reaction giving a
strongly positive reading in the presence of a non-syphilitic serum ?

From time to time in these laboratories the distilled water had
been shown to be definitely acid, PH -—, which acidity was due
to various causes, principally carbon dioxide or absorption from an
atmosphere containing acid fumes. Now it is a well-known fact
that in agglutination tests, so-called ‘ pseudo-clumping’ may occur
on account of excessive acidity of the culture medium (see

371

Biggs & Park, American Journal of Medical Science, 1897; Block,
B.M.J., 1897).

Agglutination of bacteria by acids in definite concentration can
be carried out, and the phenomenon seems to depend upon the
hydrogen ion concentration. In this connection the clumping. of
bacteria in acid agglutination is analogous to the clumping of
colloidal suspensions of any kind, and the clumping or agglutina-
tion is merely a physical phenomenon, determined by the colloidal
equilibrium of the bacteria in suspension. To digress a moment--—
this acid clumping was well exemplified by the experience of one of
my assistants in Jaffa. Hehad been getting positive results in every
Widal including controls performed in the laboratory there; and on
enquiry being made it was discovered that the new laboratory
attendant had rinsed the agglutination tubes after immersion in
acid only perfunctorily. Reasoning that a similar or analogous
phenomenon could occur in the performance of the Sachs-Georgi
test, I had certain experiments carried out which seem to prove the
likelihood of the supposition.

(a) Twelve sera were examined by the Wassermann and Sachs-
Georgi tests. The diluent of antigen and serum was normal saline
with a reaction of PH 7.

A third series was put up for the Sachs-Georgi test, but in this
series the diluent of antigen and serum gave a reaction of PH 5.

The reading of the three series at the end of eighteen hours are

as under :—
Taser VIII.
Sachs-Georgi reaction Sachs-Georgi reaction
No. Wassermann reaction (Normal saline used, PH. 7) (Saline used, PH. 5)
I XXX XXX XXX
x XXX XXX XXX
3 fo) ° XX
4 ° ° XX
5 ce) fo) fe)
6 ° fo) D4
7 XXX Xxx XXX
8 | fe) ° xX
9 ° Oo X
fo) x x xX
II fe) fe) °
12 ° ° XX
control

372

(6) In the next experiment serum was completely omitted and
the Sachs-Georgi antigen prepared with saline diluents showing a
PH reaction of 5, 6°6, 7, and 8°5.

Four tubes were arranged with contents as under :.—

Tube 1 contained 0’5 c.c. antigen (diluent PH 5) and | c.c.
saline (PH 5).
Tube 2 contained o'5 c.c. antigen (PH 6°6) and 1 c.c. saline

(PH 6°6).

Tube 3 contained 0'5 c.c. antigen (PH 7) and 1 c.c. saline
(PH 7).

Tube 4 contained 0°5 c.c. antigen (PH 8'5) and 1 c.c. saline
(PH 8'5).

These were placed in the water bath at 37° C., and read at the
end of eighteen hours. 3

Tube 1 showed marked flocculation and precipitation, while
tubes 2, 3 and 4 remained without change.

The PH reaction of the saline here is usually 6°6, and it is found
that this reaction in no wise interferes with the performance of the
Lest.

(3) A phenomenon which forced itself early on my notice was
that whereas a well marked positive reaction might be obtained in
dilutions of patient’s serum I in 20 and I in 40, no reaction
whatever was visible in the primary dilutions 1 in 5 and 1 in 10 when
the readings were made at the end of one hour, six, eighteen, and
twenty-four hours. The occurrence was relatively frequent and
demanded some explanation.

Here again one was compelled to seek a parallel in the ordinary
agglutinations in bacteriology. And an analogy certainly exists.
It must have been the experience of every bacteriologist in the
reading of results of ordinary routine agglutinations to note that
when an organism is set up against increasing dilutions of patient’s
serum, it occasionally happens that the serum in low dilution or
greater concentration fails to agglutinate the organism, while with
the serum in higher dilution or less concentration marked agglutina-
tion occurs.

This phenomenon in bacteriology has been accounted for
theoretically by the ‘ pro-agglutinoid zone,’ and the terms ‘ zones of

no reaction’ and ‘zones of inhibition’ have been applied to those
dilutions wherein agglutination fails,

373

Briefly the pro-agglutinoid theory consists in the belief that for
various reasons (é.g., length of time elapsing between the collection
and examination of the blood), the agglutinins called forth by any
specific agglutinogen may deteriorate or become converted into
substances capable of uniting with the agglutinogens without,
however, resultant agglutination.

These substances have stronger affinity for the agglutinogen than
the agglutinins themselves, and are termed ‘ pro-agglutinoids.’ If
these substances, then, are present in large numbers in strongly
reacting sera, they may wholly mask the reaction by preventing the
actual combination of agglutinogen and agglutinin. If, on the
other hand, the serum is less concentrated, then in proportion is the
number of these substances so lessened that they cannot have any
appreciable effect in preventing agglutinin from. uniting with
agglutinogen, and therefore the reaction is not obscured.

It is not, perhaps, logical to strain the similarity between the
two phenomena too far when it 1s to be remembered that the Sachs-
Georg! reaction is not even a specific antigen-antibody one, the
antigen being a homogeneous suspension of lipoidal substances, and
the antibody (which bears probably no relation to true antibody),
a lipotrophic substance 1n syphilitic serum.

Similar phenomena, however, have been observed with non-
specific agglutinating agents, and also in the action of coagulating
agents on colloid emulsions.

Orthophosphoric acid, for example, will agglutinate a certain
volume of a suspension of B&. colz when present to the extent
of between 118 cgrm. and 4 cgrm., and between 1°1 mgrm. and
O'OOI mgrm., but not in intermediate amounts between 40. and
I'l mgrm. (Hewlett).

Again, certain chemical substances have the power to agglutinate
organisms, although their action is in no way specific, and the same
substances will agglutinate different organisms (Beco). A mixture
of equal parts of commercial formalin, alcohol, hydrogen peroxide,
a I in 1,000 solution of chrysoidin, vesuvin, safranin, or perchloride
of mercury, agglutinates the typhoid bacillus as well as other
organisms. Whether, then, the phenomenon of zones of inhibition
in the Sachs-Georgi reaction is determined by physical, chemical

or other changes in the serum is not yet understood, but the analogy

374

between this and the phenomena occurring in ordinary routine
agglutinations is at least very striking.

It is obvious, then, that if reliance were to be placed on the
readings of the lower dilutions, or that if lower dilutions only
(e.g., I in § and 1 in 10) were to be put up, the results would be
untrustworthy.

(E) CONCLUSIONS AND INFERENCES

1. The Sachs-Georgi cannot take the place of the Wassermann
reaction, and should not be employed alone unless it is impossible
to obtain the reagents necessary for the performance of the
Wassermann.

2. The advantages claimed for the Sachs-Georgi reaction
are :—

(a) Negligible cost of reagents and necessities.

(6) Simplicity of technique.

3. The Sachs-Georgi, from its percentage of agreement with
the Wassermann Reaction (86-89), has a quite definite value, and if
strict attention be paid by laboratory workers to the following
points—fallacies (which constitute the main disadvantages of the
reaction) may be largely obviated, and the Sachs-Georgi may be
considered at least a useful aid in the diagnosis of syphilis :—

(a) The patient’s serum must be as fresh as possible and free
from organismal contamination. If doubt exists as to
sterility, cultural tests should be applied. (in this
laboratory it has become routine practice to inoculate
culture media tubes during the time the reactions are
being perfornied, from all cases where the sera have been
submitted from a distance and which might be likely
to be contaminated. If growth occurs, the result is
discarded.)

(6) No opinion should be given on the results of this reaction
unless a detailed history of the case accompanies the
serum—this with a view to exclude the co-existence of
other diseases. The presence of organisms, or their

375

products, in the patient’s blood may completely negative
the value of the reaction. In cases where a patient is
suffering from an acute infectious fever, the reaction
should not be employed.

(c) The saline diluent of antigen and patient’s serum must
be freshly prepared, and its reaction very carefully
estimated. Its reaction to PH should be 7, and a varia-
tion of not more than 6°6 to 7 allowed.

(2) Not less than 3 (preferably 4) dilutions of patient’s serum
should be put up in each series, the last tube of
the series showing a dilution of not less than I in 40-——
this to obviate the fallacy dependant on the ‘zones of
no-reaction.’

(€) Final opinions should not be given until eighteen to
twenty-four hours have elapsed from the time the
reaction has been performed. Pseudo-flocculation, which
may occur during the first few hours, tends to disappear
before the end of twenty-four hours and, if still present,
can be dispelled by gentle agitation of the tubes.

4. The treated cases which have been controlled throughout the
full course tend to show that the Wassermann reaction remains
positive longer than the Sachs-Georgi. A negative Sachs-Georg1
reaction, then, in treated cases would not form a reliable index as to
cure of the patient.

5. The Sachs-Georgi reaction remains negative in certain
definitely established cases of syphilis, and this for no apparent
reason. It is justifiable, therefore, perhaps to conclude that a
negative Sachs-Georgi reaction is of little or no value.

My thanks are due to Col. G. W. Heron, D.S.O., Director of
Health, for his unfailing encouragement; to my colleague, Dr. R.
Briercliffe, O.B.E., for controlling those readings which may be
regarded as controversial; and to Dr. K. Krikorian and Mr. K.
Daghlian, of the Central Laboratory, whose assistance and lively
interest in the work have rendered the production of this article

possible.

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377

NOTE ON AEDINUS AMAZONENSIS,
LuTz

A, Me EVANS, M.Sc:

(Received for publication 1 August, 1923)

Amongst a consignment of mosquitoes collected on board
s.s. ‘Hildebrand’ by Dr. A. Aiken Clark, during a voyage up the
Amazon to Manaos, in 1922, was ‘a male specimen of Culex with
reduced palpi. It appears to be closely related to Culex (Carrollia)
paraplesia, Dyar (1922), and comparison with the description of
Aédinus amazonensts, Lutz, suggests that it belongs to this little-
known species.

Culex, sp. incert.

MALE. Head. Occiput with narrow curved creamy scales
intermixed with pale golden upright forked ones above, with flat
white scales at sides below and coarse golden setae projecting
forward round eye margins. Palpi slender, about one-seventh the
length of the proboscis, vestiture of pale brown scales. Proboscis
bent at outer two-thirds, expanding distally (these two conditions
probably due to accident), scales blackish brown, labellae yellowish.
Antennae densely plumose, hairs of whorls blackish brown.

Thorax. Prothoracic lobes pale ochraceous, with a few flat
whitish scales above and a row of blackish setae. Mesonotum with
integument bright ochraceous, irridescent ; in certain lights a narrow
median dark stripe visible; vestiture of hair-like, bronze-coloured
scales, with paler reflections, the scales in front of ante-scutellar
space paler. Scutellum pale brown, with pale scales.

Abdomen. WDorsally blackish-brown scaled, with minute creamy
triangular lateral spots on the last two segments. Venter largely
denuded.

Legs with vestiture of blackish-brown scales.

Wings with first fork cell long and narrow, about four times as
long as its petiole; second fork cell about twice the length of its

378

petiole. Scales on distal half of wing mostly of the type illustrated
in the accompanying figure (fig. 1A) widest beyond the middle,
and with rounded apex. Scales on proximal half short, broad and
truncated.

Hypopygium (fig. 2). Side-pieces (fig. 2 A) closely resembling
those of C. (Carrollia) paraplesia, Dyar, but with a greater number
of stout spines between the lobe (/.) and the clasper; apical part
of lobe missing. Mesosome (ig. 2 B, C and D): lower bridge with
small, highly chitinised, finely setose area (fig. 2D, 4.). Halves
of mesosome consisting of thin plates of the form shown in the
figures (fig. 2 Band cC). In dorsal aspect the proximal portion (/.)
is seen to give rise to a short inner portion (z.) and a much longer
outer portion (0.), which constitutes the main part of the mesosome
plate as seen im lateral view. Nzxth tergites (fig. 2 E) about twice
as high as broad; ¢exth sternites comb-shaped, with eight teeth.

It has recently been suggested by Dyar (1923) that the specimens
described as Culex originator, Gordon and Evans (1922), from the
Amazon Region, represent Aédizmus amazonensis, Lutz. I have,
therefore, compared the types of C. ovzginator with the description
of Lutz’s species and find that they differ in the colour of the
thoracic integument, which is dark grey as described, and not
ochraceous as in A. amazonensis; and in the character of the scales
of the mesonotum and wings. The thoracic scales are narrow, but
not hair-like as they are said to be in A. amazonensis, and there are
no scales on the wings which could be described as ‘ Taeniorhynchus-
like.’ The main types of scales found on the apical part of the
wing are illustrated in fig. 1; the scales on the proximal half of the
wing are short and truncated. The specimen described above,
however, agrees with A. amazonensis in these three particulars, the
thoracic integument being of a conspicuously ochraceous colour.
The only noteworthy difference appears to be the absence of a well
defined median and fainter lateral dark mesonotal stripes in my
specimen, which only shows a faint median stripe in certain lights.
If, however, this character be subject to variation, it would appear
highly probable that this Culex 1s A. amazonensis.

The structure of the side-pieces would seem to indicate a very
close relationship with Culea (Carrollia) paraplesia, Dyar, but the
latter differs in other hypopygial characters, the tenth sternites

0:05 Millimelei:

Fic.1. Scales from upper branch of vein II. A.—Culex sp. incert ; B.—Culex originator,
Gordon and Evans. d.—scales of upper surface of wing.

}--—— -—- --
0:05 Pillimeler:

Fic. 2. Culex sp.incert. Hypopygium. A.—Side piece, in part with clasper. /.—lobe,
broken distally. B.—Mesosome from above. 4.—horns; 7.—inner, 0.—outer, b.—lower
bridge.

380

having only three or four teeth, and the ninth tergites being
undeveloped. |

It is thus seen that at least two species of Culex with reduced
male palpi occur in the Amazon Region; possibly others may be
discovered. Should one be found agreeing externally with
A. amazonensis, and having marked thoracic stripes as in that
species, I would suggest that the name Culex hildebrandi be used
to designate the species described above.

REFERENCES

Dyar, H. G. (1922). Notes on Tropical American Mosquitoes (Diptera, Culicidae). Ins. Ins.
Mens., Vol. X, Nos. 10-12, p. 192.

(1923). Notes on Goeldia. (Diptera, Culicidae). Ins. Ins. Mens., Vol. XI, Nos. 4-6,
p- 82.

Gorpon, R. M., and Evans, A. M. (1922). Mosquitoes collected in the Manaos Region of the
Amazon. Ann. Trop. Med. & Parasitol., Vol. XVI, No. 3, p. 323.

Lutz, A. (1905). Novas especies de mosquitoes do Brazil. Imprensa Medica.

381

THE TREATMENT OF AMOEBIC
DYSENTERY

BY
Rk. M. GORDON

(Received for publication 15th August, 1923)

The majority of the cases recorded in the following paper were treated
as in-patients at the Tropical Ward of the Royal Infirmary, Liverpool,
their subsequent history after discharge being followed at the Tropical
Clinic in the same city. At first it was hoped that much information
might be gained by consulting the hospital and other records of the
Ministry of Pensions, and through the courtesy of Dr. Finlay and others
some four hundred case sheets of amoebic dysentery patients were
examined. The results obtained were disappointingly meagre ; a few of
the cases are included in the tables that follow, but in the majority of
instances the observation periods after treatment were too short to test
the value of the drug given.

The following definitions were adhered to throughout :—

(1) The first diagnosis of amoebic dysentery was made by the finding
of motile amoebae containing red cells in the faeces.

(2) Such a patient was considered to have relapsed after the completion
of treatment when diarrhoea again occurred and active amoebae were
observed in the stool, blood and mucus being usually, though not always,
present.

(3) Cases which after treatment passed F. histolytica cysts unaccom-
panied by motile amoebae were not considered to have relapsed, but all
such instances are recorded in the tables under the heading “* Remarks.”’

(4) Once a patient relapsed he was considered as a “ fresh case ”’
and any other course of treatment was placed under a separate entry.

While undergoing treatment and for the first fortnight after treat-
ment the stools were usually examined twice weekly (sometimes, as in the
case of the emetine periodide series, much oftener) during the remainder
of the observation period the examinations averaged about one a fortnight.
In every instance, tests for the presence or absence of amoebae in the

382

stools were made by some member of the Liverpool School of Tropical
Medicine. |

No attempt has been made to compare the value of any two forms
of treatment, as the observation period was not constant. Thus the
relapses in treatments I and II were respectively 84 and 75 per cent.,
but if we fix an arbitrary limit of one month’s observation and disregard
all relapses occurring at a later date, then the relapses become respectively
53 and 17 per cent.

Note on cases treated with emetine periodide. Willmore (1923) records
ninety-one cases of amoebic dysentery treated with emetine periodide
of whom forty-eight (52 per cent.) relapsed. His observation
period is similar to that used in the present paper, but his definition
of a relapse includes persons passing FE. histolytica cysts after
treatment. Applying this definition to the sixteen cases recorded in
Table I, treatment No. IX shows that ten of the sixteen cases
(62 per cent) relapsed. Various vehicles for administering the drug
were tried; formalised gelatin capsules were given in two cases
of the acute type and all the motions passed in the subsequent twenty-four
hours saved, by this method it was found that in both instances the
capsules were passing through intact; even the plain gelatin capsules
administered to patients with diarrhoea frequently passed through the
gut without dissolving. Rice paper cachets were excellent, but owing
to their brittle character sometimes allowed part of their contents to
escape. At present we give the drug mixed with a little milk; taken
this way it sometimes causes slight nausea but never vomiting. As
Willmore’s cases were all of the type that had ‘ proved refractory to all
the known standard methods of anti-amoebic treatment’ it appears of
interest to record two cases of acute dysentery, one which (H.J.C.) had
never received previous emetine treatment. Amoebae disappeared from
this man’s stool within forty-eight hours of the start of treatment and did
not reappear during the six weeks he was kept under observation. The
other case (C.M.P.) had received only one previous course of emetine
(twelve grains emetine hydrochloride given fifteen months previously).
During the first six days of treatment this patient continued to pass
blood, mucus and amoebae, and at the end of this time his condition
was so bad that it was thought advisable to supplement the periodide
with four hypodermic injections of emetine hydrochloride. Under this
combined treatment the amoebae vanished from the stools within twenty-

———

Nature of treatment

‘#TMENT No. I.
Showing the effect of
ment with emetine

ychloride gr. 1 given sub-
Saal or intramuscularly
ro to six consecutive days.

rMENT No. II.

thowing the effect of
nent with emetine
chloride gr. 1 given sub-
eously or intramuscularly
a to fourteen consecutive

‘MENT No. III.

howing the effect of
aent with emetine
th iodide grs. 3 given by
1 on twelve or thirteen
tutive days.

the effect of

with —emetine
chloride gr. 1 given sub-
sously together with
1€ bismuth iodide gr. 1
by the mouth on twelve
utive days. (Three cases
en days).

Relapsing

Not
relapsing

Relapsing

Not
relapsing

Relapsing

Not
relapsing

Relapsing

Not
Relapsing

383

TaBLe I.

Observation period in months after
completion of treatment

E 2 3 4 5 cen
| }
7 a .
| '
}
| = —_—_—
} |
|
I EE ‘
— :
| |
5 4 I 3 : 2
I 2 . 2 ae
!
IO 4 3 i 2 ‘
3 2 I Bes +4 a
I I AA = we ys, ae
| '
|

Showing the effect of various forms of treatment on one hundred and thirty-eight cases of amoebic dysentery.

Total
Remarks

One non-relapsing case was
passing E. histolytica cysts four
months after completion of
treatment.

Most of the cases in this series
were of the acute type, i.e.
passing a large number of blood
and mucus motions in the
24 hours.

ST ES

One non-relapsing case was
passing £E. histolytica cysts
12 months after completion of
treatment. The majority of
the cases were of the acute
type. Two non-relapsing cases
7 | were observed for more than

a year.

Most of these cases were of
the chronic type, i.e. passing
a daily average of four to six
loose stools containing active
7 | amoebae but little or no blood
or mucus.

Two non-relapsing cases
were passing E. histolytica cysts,
respectively, 12 and 20 months
after completion of treatment.
The cases include about an
equal proportion of acute and
chronic types. Three non-
8 | relapsing cases were observed

for 18 months.

384

Taste I.—continued.

Showing the effect of various forms of treatment on one hundred and thirty-eight cases of amoebic dysentery.

Nature of Treatment

TREATMENT No. V.

Showing the effect of
treatment with pulv. ipecac.
ers. 5, together with pulv.
ipecac. Co. grs. 5 given by
mouth on thirty consecutive
days.

‘TREATMENT No. VI.

Showing the effect of
treatment with pulv. ipecac.
grs. 5, together with pulv.
ipecac. Co. grs. § given by
mouth on sixty consecutive
days.

Treatment No. VII.

Showing the effect of
treatment with Ravauts paste,
three drachms given by mouth
on thirty consecutive days.

TREATMENT No. VIII.

Showing the effect of
treatment with Ravauts paste,
three drachms given by mouth
on sixty consecutive days.

Relapsing

relapsing

Relapsing

Not
relapsing

Relapsing

Not
relapsing

Relapsing

Not
relapsing

TREATMENT No. IX.

Showing the effect of
treatment with emetine
periodide grs. 6 given by

mouth on thirteen to fifteen
consecutive days. In one case,
which is included amongst the
non-relapsing, the emetine
periodide was supplemented
with four injections of emetine
hydrochloride gr. 1.

Relapsing

Not
relapsing

Observation period in months after

completion of treatment.

Ss)

3 7
I
| fi 4
if

|
8
8

Remarks

| |

:
The majority of |
were of the chronic type.

Most of these patients
of the chronic type andi
repeatedly relapsed — u}
various other forms of t
ment.

These were mild case
a chronic type.

These were mild case

a chronic type.

Two non-relapsing cases
passing E. bistolytica
respectively, two and six Y
after the completion of t
ment. The cases include a
an equal proportion of 4
and chronic types.

bas

385

four hours and no relapse occurred during the five months the patient
was kept under observation. Two other cases who had received numerous
previous courses of emetine continued to pass motile amoebae throughout
the time of treatment. Daily examinations of the remaining twelve
cases showed that in two of them motile amoebae persisted for five days
of treatment and in the other ten cases vanished after one to three days.

Taste II.

Showing effects of various treatments not recorded in Table I.

Number
Treatment. of cases Result of treatment.
treated.
Emetine hydrochloride $ gr. given subcutaneously on
two consecutive days. I Relapsed after six days.
a a EC a eee —_ ee ———
Emetine hydrochloride 4 gr. given subcutaneously on
twenty-two consecutive days. | I Relapsed seven months later.
Emetine hydrochloride } gr. given subcutaneously on |
thirty consecutive days. | 1 Relapsed within a month.
Emetine hydrochloride gr. 1 given subcutaneously | (1) No relapse after three
together with emetine hydrochloride $ gr. given by mouth 2 months observation.
on ten consecutive days. (2) No relapse after three
months observation.
Emetine hydrochloride gr. 1 given subcutaneously on (1) No relapse after twelve
six consecutive days followed by emetine bismuth iodide 2 months observation.
grs. 3 on twelve consecutive days. (2) Relapsed six months later.
Emetine hydrochloride gr. 1 given subcutaneously on |
twelve consecutive days followed by emetine bismuth I No relapse after twelve
lodode grs. 3 given by mouth on six consecutive days. months observation.
Emetine hydrochloride gr. 1 given subcutaneously | (1) No relapse after six weeks
together with emetine bismuth iodide grs. 3 given by | 2 observation.
mouth on six consecutive days. (2) Relapsed a week later.
Emetine bismuth iodide gr. 1 given by mouth on |
twenty-four consecutive days. / I Relapsed within a month.
‘““Yatren’”’ 200 ccs. of a 5 per cent. solution given
per rectum on ten consecutive days, then six days rest I Relapsed within a fortnight.

|
followed by a like dose for one day only. |
|

Note on case treated with Yatren. Mihlens and Menk (1921) recommend
ten grammes of Yatren given by the rectum for eight to fourteen days,
then no treatment for seven days ; repeat the Yatren for three to seven
days, allow another resting period of seven days and repeat treatment
for three to five days. Owing to the fact that only a limited quantity of
the drug was available, the course was shortened to that shown in Table IT.

386

‘Taser III.

Showing the distribution in various months of the numbers and percentages of one hundred and one
relapses after treatment.

Month in which relapse occurred
after completion of treatment.

Treatment. Relapses._ --—— —— ——- ————

More
Zo} Ql 034k ok ON etian

Emetine hydrochloride gr. 1 given subcutaneously
or intramuscularly on two to six consecutive days.

Emetine hydrochloride gr. 1 given subcutaneously
or intramuscularly on ten to fourteen consecutive
days.

Percentage] 23 |

Emetine bismuth iodide grs. 3 given by mouth on [Number [10] 4)| 3]... 2/.. 6
twelve to thirteen consecutive days. 2 [~————{—— ——
Percentage

Emetine hydrochloride gr. 1 given subcutaneously
together with emetine bismuth iodide gr. 1 given
by mouth on twelve consecutive days (three cases
only ten days).

Number. [11 | ss. loh?aiideleneed | Salk eae

Pulv. ipecac. grs. 5 together with pulv. ipecac.
Co. grs. § given by mouth on thirty consecutive days.

Puly. ipecac. grs. 5 together with pulv. ipecac.
Co. grs. 5 given by mouth on sixty consecutive days.

Ravauts paste three drachms given by mouth on
thirty consecutive days.

Ravauts paste three drachms given by mouth on | Number 3 Wh ee fee ee I
sixty consecutive days. —-——— SS

Percentage] 75 | ...-| +

Emetine periodide grs. 6 given by mouth on }| Number
thirteen to fifteen consecutive days (One case [-—
supplemented with four injections of emetine
hydrochloride gr. 1).

Various treatments recorded in Table IT.

:
-

387

The patient (A.G.) was a chronic case of about five years’ duration who
had completely resisted, or else relapsed shortly after, numerous forms
of treatment. Before treatment commenced he was passing eight to ten
motions a day containing blood, mucus and active amoebae. Twenty-four
hours after the first rectal injection the amoebae disappeared and the
number of stools were reduced to one or two a day. This condition lasted
for twenty-eight days when blood, mucus and amoebae again appeared
in the faeces. A further supply of the drug has been obtained and other
patients are now under treatment.”

SUMMARY

One hundred and fifty cases of amoebic dysentery were given various
forms of treatment and subsequently kept under observation for one
to six months or longer ; of these one hundred and fifty cases, one hundred
and one (66 per cent.) relapsed, the numbers and percentages of the
relapses occurring in various months being recorded in the tables.
Amongst all the cases treated only six (4 per cent.) were observed
to be passing E. histolytica cysts after treatment. Sixteen cases were
given emetine periodide ers. 6 daily, eight of these (50 per cent.) relapsed
within one month ; the giving of this drug in gelatin capsules was found
to be unsatisfactory as they frequently passed through the gut without
dissolving ; the periodide when mixed with a little milk and given by the
mouth did not produce vomiting. Owing to the inequality of the observa-
tion periods no attempt was made to compare the value of any two forms
of treatment.

REFERENCES

Miutens, V. P., and Menx, W. (1921). Behandlungsversuche der chronischen Amoebenruhr mit
Yatren. Miinch. Med. Woch, Vol. LXVIII, p. 802.

Wittmore, J. G. (1923). The treatment of refractory cases of amoebic dysentery. Trans. Roy.
Soc. Trop. Med. and Hyg. Vol. XVII, p. 13.

*Since the publication of this note another case has relapsed, fourteen days after the
completion of treatment. The patient in this instance had received the full course of
treatment recommended by Miihlens and Menk.

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389

RELAPSING FEVER IN THE GOLD
COAST

bo ow. woe LWYN-CLARKE,, M.C., M.D,
Femme LerN Lie Ga
AND

A. INGRAM, M.D., C.M.

WEST AFRICAN MEDICAL STAFF
(Received for publication 20 August, 1923)

The following account of an outbreak of relapsing fever in Accra,
Gold Coast Colony, has been considered worthy of record since the
disease has never before been recognised in the Colony.

Doubts have been expressed as to the possibility of the disease
having been prevalent in the Colony in former years. For it is
well known that circumstances tend to render the recognition of the
disease unusually difficult since attacks may be so mild in character
as to resemble slight attacks of malaria. Examples of this nature
were met with during the outbreak to be described, and the danger
of spread resulting from the movements of persons suffering from the
ambulatory type of spirillar fever was readily appreciated. It is
difficult, however, to understand how the disease could have existed
undetected for, as can easily be understood, routine blood examina-
tions in all cases of fever can be numbered in their hundreds every
year, and had a proportion of these unclassified fever patients been
suffering from relapsing fever, the spironema could scarcely have
escaped detection.

In connection with this it should be stated that Simpson
discovered spironemata in a single blood smear out of a large number
of blood smears examined in 1908, the year of an outbreak of plague
in the Gold Coast Colony.

EPIDEMIOLOGY

Two possible sources of infection present themselves in connection
with the epidemic to be described hereafter. One of these appears
to have been the soldiers who returned to the colony after hostilities
had come to an end in what is now called Kenya Colony. It 1s
recorded that the West African forces, especially those in the

582

Dar-es-Salaam area, suffered severely from relapsing fever. It is
possible that a certain number remained infective subsequent to
their return to the colony and that infection spread to their families
and to neighbouring tribes.

This theory is supported to some extent by the fact that the
majority of the returned troops were members of the Northern
Territory tribes and that many of these took their discharge from
the Headquarters of the West African Frontier Force at Coomassie.
The theory gains further support from the large preponderance of
Northern Territory tribesmen, some of whom had seen war service,
amongst the patients encountered during the outbreak at Accra.
Moreover, a report has recently come to hand from the Medical
Authorities in the Northern Territories of the Colony, to the effect
that six cases of relapsing fever (confirmed by blood examination)
have been isolated from recruits to the West African Frontier Force.

There are, however, several excellent objections to the acceptance
of this theory, not the least of which is the fact that the West African
troops while in East Africa suffered from infection by Sp. duttonz
carried by the Ornithodorus moubata, whereas the strain met with
in the Accra epidemic resembled Sf. obermeieri, in this instance
the vector being the body louse.

The other possible source of infection may have been the
neighbouring French territory, since a very extensive epidemic of
relapsing fever was reported from Senegal and French Niger
Territory in 1921. |

Epidemics of relapsing fever are said to be slow in onset and
to show a gradually increasing mortality. In the outbreak under
review the onset was certainly rapid though its rapidity is slightly
obscured in the chart (No. 1) owing to the artificial conditions
resulting from emergency legislation. |The mortality, moreover,
fortunately showed no gradual increase in severity, as seen by the
fact that the case mortality rate for the first fifty cases was 6 per
cent., while that for the remainder was less than 1 per cent.

INCIDENCE
Cages

As in India and other countries where the disease is endemic
the large majority of cases occurred in adult males, the actual
figures being one hundred and fifty-six male cases (including two
European cases) and only two female cases.

391

This very large preponderance of male cases is not dependent
upon the degree of lousiness of the two sexes, for observations
showed that infestation was shared in equal measure by both men
and women. The explanation, rather, lies in the fact that the
infected males belonged almost exclusively to an immigrant tribe,
who had come into the Accra district from their Northern Territory
villages in search of work and money to purchase European goods.
It is contrary to the customs and habits of these tribesmen to bring
their women and children with them, and as they return to their
native villages as soon as they have been able to collect together a
small quantity of trade goods, they do not possess property and
housing accommodation in Accra. It is not difficult to understand
the conditions under which these people live in Accra, crowded
together in insanitary hovels lacking in light and air.

(6) Age

Since the majority of cases of relapsing fever occurred amongst
males of Northern Territory tribes, it follows that most of the cases
would occur in adults, since the journey to Accra from the Northern
Territories is not a thing to be lightly undertaken by persons
other than healthy adults. The age of the patients varied from
IO years in a Hausa boy to 55 in a Zabramah man. The patients
were almost all illiterate and consequently their ages could only be
estimated approximately—the average being between 25-30.

The following table gives the racial incidence.

Taste I,
Race or Tribe Number of Cases Percentage
European ... it 7 ae ase A a at 25 ‘2
Kroo saa & tr Be i Ah 3 7, 2t 12
Kotokoli ... 2 roe +3 ee A Doe wae 3 1°8
Hausa a? es hs dee 4 eh Pes Fr 3 1°8
Other Tribes... i soe es By ae res gt 5°6
Zabramah (Zaberrima) ... hanes ae By ni ty 139 87°9
Total ee tes ae 5h ae ey 158 99°5

* A European who became infected in the course of experiments, and who became infected
a second time at a later date, is included in this figure.

t Two volunteers who were infected in the course of experiments.

t Includes a volunteer who was infected in the course of experiments as in the other three
cases,

The reasons for the preponderance of Zabramah and _ other
Northern Territory tribesmen amongst the cases are not hard to seek.
Reports of extensive epidemics of doubtful character are not
infrequently received from the Northern Territories and one such,
accompanied by heavy mortality and attributed to cerebro-spinal
meningitis took place in 1920. It is possible that relapsing fever
may have been the cause of certain of these outbreaks and that
shortage of medical staff allowed it to remain unrecognised. In any
case an undoubted epidemic of relapsing fever occurred amongst
natives in French Territory bordering upon the Northern Territories
in 1921, and intercommunication across the frontier would account
for infection.

Once infected, the habits of these tribes would ensure a rapid
spread and perpetuation of the infection. Owing to a great scarcity
of water in most parts of the Northern Territories, except in the wet
season, the average tribesman from this area 1s brought up from
birth in the belief that water is intended for drinking and cooking
purposes only. In consequence his body, clothes, bedding and hving
quarters are quite innocent of soap and water.

This condition of affairs contributes to a state of lousiness, and
infestation with lice is so general (100 per cent. of the Northern
Territory patients admitted to the Contagious Diseases Hospital,
Accra, were found to be lousy on admission) that little, if any,
attempt at disinfestation is made on the part of sufferers from lice.
Coastal tribes in the Contagious Diseases Hospital were amazed at
the Health Authorities interfering with what was considered a
‘custom of the country’ when disinfestation of admissions was
carried out. The fact that Northern Territory tribesmen and
Hausas sleep in their work-a-day garments tends to add to the
degree of natural lousiness.

In this connection it is a remarkable fact that coastal tribes,
not excepting the Kroos, are singularly free from lice, due, no
doubt, to the fact that they do not share the aversion from washing
their bodies and clothes exhibited by Northern Territory tribesmen.
In order to obviate the possibility of the importation of plague,
smallpox and other infectious and contagious diseases into the
Colony, all Kroo immigrants from the Kroo coast are medically
examined on arrival at this port. Nine hundred and thirty-one were

393

so examined in 1922, and although they had been in most cases
crowded together on board ship and had not had facilities for
washing their bodies and clothes for as long as three weeks in some
cases, not a single one was found to be lousy.

A further reason for the large proportion of Northern Territory
tribesmen among the cases of spirillar fever les in the insanitary
conditions in which they lived in Accra. Not possessing any house
property or relatives with satisfactory living accommodation, they
crowd together in insanitary corrugated-iron structures lacking light
and ventilation, intended by their unscrupulous owners not as living
quarters but as stores for building materials and the like.

Lastly, owing to a temporary trade depression many of the
Zabramahs were unemployed for some time prior to the commence-
ment of the outbreak and, having neither friends nor relations in
the district, they became half starved. The synonym ‘ famine
fever’ indicates the traditional association of semi-starvation and
relapsing fever. With their powers of resistance so diminished they
afforded fertile soil for the germ of any infectious disease.

The number of cases occurring amongst other tribes was too small
to warrant conclusions being drawn as to the comparative severity
of the disease, but it can be stated with fairness that the more severe
type of case—and in fact all the fatal cases—occurred amongst
Zabramahs.

(2d) Case Distribution

A map is appended showing the districts in which patients
appeared to have been infected.

Neglecting the first case which occurred in a European who had
passed through a considerable number of bush villages in the Accra
District during the course of his work, a number of the earlier
patients appeared from information obtained from them to have been
infected either in their native villages in the Northern Territories,
or in one of the towns or villages at which they rested at night
time, during their journey to Accra. This bears out the theory that
the epidemic in Accra and District originated directly in Northern
Territory tribes.

It will be seen from the map that the vast majority of cases
occurred in the Tudu and Zongo Rd. areas of Accra, in both of
which the inhabitants are almost entirely members of Northern

394

Taste II.

Table to show location of places where patients are presumed to have been infected.

Number of Patients
Place infected

Accra aa ais “ vr Se rae aes Ar oS hoe 130*

Some village in Accra bush or on the road from the Northern Territories

toAccra ya.. Ate mee a ae ae a oe io 19f
Nsawam ... a ve 5 a sds AoA Ae 48 ma ae 6
Mangoase... * ais ss hie = ue 3 fs bes 3
158

*Includes three African patients accidentally infected in the Colonial Hospital, and
three Africans and one European infected experimentally at the Medical Research Institute
(the European suffered from two attacks).

t Includes one European case infected in some unknown village in the Accra bush.

Taste IIT.

Table to show areas in which cases of relapsing fever occurred, but not necessarily where
infection took place.

Number of Cases

Accra, Block No. 11 an bee oe és sea pte hs re I
Accra, Block No. 12 — ... oe tis an cpa atk RE P. 19 i
Accra, Block No. 13a... Ane ase a igs ee a0 4 43 .
Accra, Block No. 15a... es oe as ror mit, +e ere . 8
Accra, Block No. 15b_... a ma ons oye saa IF ae 50
Actra/Block Nown6yaetliy decel.do tetfliame stelephtanos 2 We 13
Accra, Block No. 17... 1% die ore ae MS teh if 3
Accra, Medical Research Institute ... vas arp at tare as 4*
Accra, Native Hospital «77 J occca 9) gas anne ee cs cE 3
Accra Bush Se Pte Bes: ate date ee 20% a4 mS 6+
Residence unknown wen £, ssf sy » ode cae fi 8

Total ose ness 5 dhe rey 158

* These cases were infected experimentally.

_t This number includes a European, the first case to be discovered at the commencement
of the epidemic. nr

\
—AccRA—

\
70 SHOW WHERE CASES OF FIELAPSING FEVER —

NLURRED DURMG THt LUC. —

— Marcnw ro/uly 1923 ——

CB
Xs

OY — Scate 1i2z500 —

396

Territory tribes or Hausas—both of whom live under unhealthy
conditions.

Four cases are shown to have been infected during the course of
experiments aiming at the discovery of the means of transmission
of the spironema, while three cases are shown as having been infected
in the general wards of the Native Hospital, in which a number of
relapsing fever patients were treated, prior to the declaration of the
disease as an infectious disease necessitating removal to the
Contagious Diseases Hospital.

It is noteworthy in connection with the distribution of cases that
the areas from which they came were of a very low standard as
regards housing accommodation, overcrowding in insanitary, ill-
lighted and ill-ventilated hovels.

(e) Mode of Transmission of the Disease

During the initial stages of the Accra outbreak of spirillar fever
preventive measures against the spread of the disease were greatly
hampered by the lack of knowledge of the exact means of trans-
mission of the organism,

Ornithodorus noubata Murray, the carrier of tick fever in the
Congo, and Ornithodorus savignyi Aud, the possible carrier of
relapsing fever in Somaliland, have not yet been recorded!
as occurring in the Gold Coast. Ovnithodorus talaje Guérin-
Meneville, the carrier of relapsing fever in Panama, was taken by
Graham on Cricetomys gambianus (Report on Plague in the Gold
Coast in 1908: W. J. Simpson, p. 22), it was also taken on
M. decumanus during the small epidemic of plague in Accra in 1917,
but its occurrence 1s so rare as to preclude its being the prevalent
carrier of the present outbreak of relapsing fever. Avgas persicus
Oken is supposed by some inhabitants of the Gold Coast to be a
common parasite of fowls in the Colony, but there is no authentic
record of its occurrence; moreover, it has been shown by Edm.
Sergent and H. Foley (1922) that this tick is not a carrier of North
African relapsing fever, whereas Pediculus humanus is an efficient
one.

Feeding experiments were carried out with bed bugs, mosquitoes,

and lice.

Bed bugs
On the 14th April seven bed bugs collected from the clothes and

397

bedding belonging to cases of relapsing fever which had _ been
interned in the Contagious Diseases Hospital, were placed on the
shaven back of a small monkey. At least two of these bugs fed on
the monkey, but it was difficult to keep them in position so that
the experiment was abandoned within four hours of its commence-
ment. The monkey during the following fortnight showed no
symptoms of illness and spironemata were never found in its blood.

A second consignment of bugs from the same source numbering
a dozen was placed on the arm of a native volunteer on the 15th
April, a modification of Nuttall’s pill-box method of feeding lice
being adopted. The bugs were retained on the arm for three days
and were then removed and dissected, no spironemata were found in
them and the volunteer never showed any symptoms of relapsing
fever.

On the 30th April twenty bugs which had been collected from
the bedding of relapsing fever cases were kept alive in the incubator
for four days and were then ground up in saline. The coarser
particles having been removed, the emulsion, which contained no
spironemata, was rubbed into the scarified skin of a volunteer;
the volunteer never showed signs of relapsing fever and spironemata
were not detected in his blood at any time during the two weeks
following the inoculation.

Mosquitoes.

Aedes argenteus Poiret (S. fasciata) being probably the most
universally distributed mosquito in West Africa, and being the
recognised carrier of Leptospira icteroides, was selected for experi-
ment. Eleven female Aedes argenteus, reared from larvae, were
placed in a gauze-covered jar and fed upon the arm of a patient
suffering from relapsing fever in the Contagious Diseases Hospital
before the patient had received any treatment. Five of the
mosquitoes were seen to be engorged with blood when they were
brought back to the laboratory on the 13th April. The mosquitoes
were separated into two lots; six mosquitoes, of which three had
certainly sucked blood, were placed in one jar and five, of which
two were engorged with blood, were placed in another jar, honey and
water were placed in these jars and the mosquitoes were kept alive
with occasional renewal of the honey until the 24th April. Two
volunteers began to feed these mosquitoes on their arms on that date

398

—(one jar being assigned to each volunteer)—and continued to feed
them daily till the 2nd May when the mosquitoes began to die. No
spironema was found in any of the surviving mosquitoes on
dissection and neither of the volunteers showed symptoms of
relapsing fever during the fortnight following the abandonment of
the experiment, and at no time were spironemata found in the blood
of either of them.

Lice. (Pediculus humanus).

On the 14th April four lice Pediculus humanus), collected from the
clothing of a case of relapsing fever, were placed by the aid of the
pill-box method on the back of a monkey, but they refused to suck
blood and were therefore transferred to the arm of a native volunteer :
on the 17th April these lice were all found to have died. They were
at once ground up in saline and the emulsion was inoculated into a
black rat. Neither the volunteer nor the black rat showed any signs
of illness and no spironemata were at any time found in the blood
of either.

‘On the 30th April twelve lice were placed on the arm of a
native volunteer; these lice had been found in the clothing of
contacts with relapsing fever cases, nine of the lice were found to be
dead on the following morning. The three surviving were then
placed on a case of relapsing fever in the Contagious Diseases
Hospital; after sucking blood from this case they were brought back
to the laboratory, kept in the incubator for forty-eight hours, and
were then placed on the arm of a second volunteer who managed
to keep them alive for three days. Neither of the volunteers suffered
in the least degree during the two weeks succeeding the feeding of
the lice and neither showed spironemata at any time in his blood.
The nine lice from this batch which were found dead on the arm of
the first volunteer on the 1st May were ground up in saline and
the resulting emulsion injected subcutaneously into a monkey
without any effect following, the monkey remained well and
spironemata were never seen in its blood.

On the 14th May a dozen lice obtained from the clothing of a
case of relapsing fever in the Contagious Diseases Hospital were
placed on the arm of a native volunteer and left iz situ for seventy-
two hours. At the end of that time ten were found dead, the two
survivors together with the ten dead were ground up in saline and

399

the emulsion inoculated subcutaneously into a monkey. Neither the
volunteer nor the monkey developed symptoms and spironemata were
not found in the blood of either during two weeks following the
experiment.

On the 16th May twenty-five lice were received from the Medical
Officer of Health who had collected them from the clothing of cases
of relapsing fever. These lice were divided into two lots and fed
on the arms of two volunteers for three days with the same result as
in previous experiments—neither volunteer developing symptoms of
relapsing fever or at any time showing spironemata in his blood.

The droppings from these lice adherent to the sides and bottom
of the test tube in which they were received were gently washed out
with saline and the mixture rubbed into the scarified arm of a
volunteer, but he developed no symptoms and his blood remained
free from spironemata for two weeks following the inoculation.

On the 26th May two dozen lice collected from the clothing of
several cases of relapsing fever in the Contagious Diseases Hospital
were received and were placed on the arm of one of us (A.I.) by the
aid of the pill-box method; these lice were fed at intervals during
the day and night till the 4th June when, there being only six
survivors, they were ground up in saline and the emulsion which
contained spironemata was inoculated into the scarified arms of two
native volunteers. On the 12th June one of the volunteers complained
of headache and lumbar pain, his temperature was found to be
100°2°F., and spironemata were found in thin and thick films of his
blood. On the 13th June the other volunteer reported sick, his
temperature was 100°6°F. and his blood showed spironemata. No
spironemata were present in the blood of the individual upon whose
arm these lice were placed at any time; it was examined from the
26th May to the 4th June whilst the lice were being fed.

On the 4th June four lice were supplied by the Medical Officer
of Health (P.S.S-C.). These lice had been allowed to remain on a
patient suffering from relapsing fever for five days after he had
been treated with Novarsenobillon. When received at the laboratory
the lice were placed on the arm of a.native volunteer using the
modified pill-box method; they were left on this individual’s arm
for twenty-four hours only and one of them had disappeared when
the pill-box was removed. The three survivors were ground up in

400

‘saline and a portion of the emulsion was rubbed into the scarified
arm of another volunteer, a second portion was dropped into the
conjunctival sac of a monkey, while the residue was inoculated
subcutaneously into a second monkey. ‘This emulsion contained
spironemata. Neither of the monkeys became ill and during the
two weeks following the experiment no spironema was found in the
blood of either. The second volunteer into whose arm the emulsion
had been rubbed suffered from headache, and had a temperature
of g9g'2°F. on the 14th June, but no spironemata were found in thick
films of his blood examined daily for a week after the slight rise
of temperature. The first volunteer upon whose arm the four lice
were placed began to be ill on the 15th June, but did not report
himself as sick until the 18th June, when his temperature was found
to be 102°F. and spironemata were numerous in thick films of his
blood.

On the 11th June twenty lice from the clothing of cases of
relapsing fever in the Contagious Diseases Hospital were supplied
by the Medical Officer of Health (P.S.S-C.); these were at once
placed on the arm of a native volunteer who fed them for three days.
On the 14th June, however, as he seemed reluctant to continue to
feed the lice, the survivors, five in number, were transferred to the
arm of one of us (A.I.) where they were fed for another four days.
During the last period of feeding two of the lice escaped one night
from under the bandage retaining the pill-box in position. and
wandered freely over the body—judging from the number and
position of the bites discovered the following morning. These lice
were not recovered and may have been crushed in an attempt to allay
the irritation of their bites. On the 18th June the feeding of the
three ultimate survivors was discontinued and they were ground up
in saline and the resulting emulsion, which contained spironemata,
was inoculated subcutaneously into a monkey. This monkey showed
no symptoms of illness and spironemata were not at any time found
in its blood, which was examined daily till the 2nd July. The
native volunteer who fed this batch of lice for three days immediately
following their last meal of infective blood remained free from
sickness, and no spironemata were found in his blood which was
examined daily for twelve days after he had discontinued feeding
the lice. The second individual who continued the feeding of the

401

lice became ill on the 25th June, his symptoms were headache and
pains in the limbs and his temperature was 101°6°F., but no
spironemata were found in a thick film of blood; on the morning
of the 26th June when the temperature was 102.4°F. a few spirone-
mata were seen in a thick film of blood, they became more numerous
at a later stage of the attack.

It may be of interest to mention that the individual who became
sick as the result of this last feeding experiment was the same who
developed relapsing fever as a result of accidental inoculation on the
28th March. It is possible that this second attack may have been
a relapse of the former, but it appears to us more probably a
reinfection, the interval between the recovery from the first attack
and the onset of the second, a period of eleven weeks, being too great ;
besides at no time during the interval were any spironemata found
in the blood which was repeatedly examined, and no symptoms of
illness were experienced.

These feeding experiments appear to agree with the conclusions
of the French observers in Tunis and Algeria, Ch. Nicolle, L. Blaziot
etl. Conseil) (1012), and-E.dm. Sergent et H. Foley, (1922),
namely, that the disease is not conveyed by the bites of lice or by
their droppings being rubbed into excoriations of the skin, but that
it is conveyed by the inoculation of crushed lice into wounds of the
skin; further, that lice must be kept alive for about one week after
feeding on a case of relapsing fever before they are capable of
conveying the infection. !

The two last experiments may seem rather equivocal, suggesting
that the infection is conveyed by the bites of lice alone. It is to be
noted, however, that in both experiments one or two lice escaped

*from the pill-box in which they were enclosed and wandered over the

body generally; it is therefore quite possible that these stray lice
were unconsciously crushed and rubbed into abrasions of the skin
made in the process of scratching.

The pill-box method of feeding may not be an ideal method for
use in the tropics, as pointed out by Cragg (1922), but when properly
applied it certainly prevents crushing of its contents on the skin by
any attempts at scratching.

Up to the time of writing, experiments have not been carried
out to show whether infection is transmitted by infected lice to their

402

eggs, though this has been shown to be the case in other parts of
the world.

It is noteworthy that lice were found on a very large proportion
of all the patients treated at the Contagious Diseases Hospital,
more especially among Zabramahs and other Northern Territory
tribes and Hausas, in whom lice were found on the hair of the head,
beard, axillae, pubic region and on wearing apparel.

The louse appeared to resemble the head and body louse found
in Europe—Pediculus humanus L. It was remarked that lice tended
to migrate from an individual having a high temperature, suggesting
that the optimum skin temperature was probably in the neighbour-
hood of 98°4°F. or less. The temperature of the air and the relative
humidity did not appear to influence the numbers or habits of the
louse.

A graph and table given in the appendix illustrates the lack of
influence exercised by temperature and humidity on the numbers of
cases discovered in Accra from week to week.

(7) Lime of Occurrence of Cases

A chart 1s appended to show the progress of the epidemic from
week to week. A somewhat erroneous impression is gained from an
inspection of the chart, however, since the peak of the outbreak would
appear to have occurred in the weekly period April 29th—May 5th.
The large excess of cases occurring during this and the following
weekly period resulted from legislation being passed on April 28th,
which allowed the Health Authorities to round up nearly four
hundred suspected cases and contacts with cases isolated from certain
areas. |

Briefly, a steady weekly increase in the number of cases reported _
occurred from the first case on March 18th, attaining a maximum
during the weekly period April 29th—May 5th, and then steadily
decreasing until only one case occurred during the weekly period
June 24th—-30th, this case being a second attack in a European
infected experimentally in order to confirm the mode of trans-
mission of infection.

(g) Meteorological

A daily maximum temperature of over 98.8°F. is stated to exert
an unfavourable effect on lice. Conclusions based upon observations

403

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carried out over a period of three months showed that the small
variations in atmospheric temperature and relative humidity had
little or no influence on the degree of infestation of the Northern
Territory tribes normally found infested with lice.

The outbreak commenced towards the end of the dry season
during March, when the average maximum shade temperature was
88°F. and the relative humidity 67.

The greatest number of cases occurred at the end of April and
at the commencement of May, during which times the rainy season
had been in progress for a short time, and the average maximum
shade temperature and relative humidity for April and May
respectively being 88°F. and 86°5°F. and 671 and 769. The
epidemic virtually came to an end early in July during the continua-
tion of a rather more than normally wet season. The slight fall in
temperature and the decided increase in rainfall between March
and July appeared to have little influence on the course of the
epidemic or upon the severity of individual cases.

(h) Morphological Characters of the Spironemata

The spironema found in the blood of cases met with in the present
epidemic differs in no way from the descriptions given of the
spironemata causing relapsing fevers in other parts of the world.
Two hundred spironemata taken as they came—twenty-five in eight
blood films from separate cases—were drawn with the help of the
camera lucida and measured by the compass method, Macfie and
York (1917). The shortest spironema found measured 10 and the
longest 44m, the average length being 21:9”. The commonest
lengths of the spironemata were 18m to 23m, and the average
thickness of the spironemata was 0°34. The pleomorphism noted by
the French observers, J. Kerrest, A. Gambier et A. Bouron (1922),
in the Soudan epidemic of relapsing fever has also been noticed by
us, but it appears to us to be merely a passing phase; in blood
films obtained from the same case on consecutive days, we have
found few, if any, irregular forms on the first day, while on the
second day ring and figure-of-eight forms have been numerous and
did not require to be searched for. Breinl (1908) states with regard
to Sp. duttoni, that coiled and complicated skein-like forms are
most numerous in the blood of the internal organs just before the
crisis sets in. Balfour and Bousfield (1911) have described and

ips)

figured these irregular forms of spironemata in relapsing fever at
Khartoum. With the exceptions of ring, figure-of-eight and partially
coiled forms, the shape of the organism did not appear to undergo
any change in patients from day to day and, although the majority
of the films examined were air-dried before being treated with
Ruge’s fluid, the exposure of blood films to hot air, to the vapours
of formalin, to osmic acid, or to chloroform, appeared to have no
influence on the shape of the organism.

The spironemata found in emulsions of crushed lice appeared to
be shorter and more delicate than those seen in blood films, they
also stained less deeply with gentian violet.

The number of organisms found in thick blood smears varied
from over forty per field observed in a case which resembled in many
respects a typical case of lobar pneumonia to as few as two over
the greater part of the slide.

It would have been anticipated that spironemata would have been
more numerous in severe cases of the disease and in first attacks than
in relapses, but this was not invariably the case, although as a general
rule they were less easy to find in relapsing cases and in fact were
rarely found in what appeared to be a relapse after injection of a
substerilising dose of Novarsenobillon. In one case spironemata
were found by Dr. Mary Magill (who kindly assisted to examine a
group of nearly two hundred films prepared from contacts and
suspected cases) in a blood film of a contact who was not suffering
from pyrexia at the time nor for the forty-eight hours intervening
between his blood being taken and his treatment with Novarseno-
billon. This case showed no signs or symptoms of illness and was
discharged fourteen days subsequent to his receiving an intravenous
injection of o°3 gm. of Novarsenobillon, not having shown any signs
of sickness. This blood film was one amongst twenty-five other
films of contacts, all of whom appeared and were healthy; thus the
possibility of the slides having become mixed could be excluded.

It is a remarkable fact that a careful search through blood films
taken from some patients who appeared to be suffering from typical
attacks of spirillar fever, who were stricken at the same time as their
comrades, and who reacted to intravenous medication in exactly the
same way as their fellow patients, failed to show the presence of

infecting organisms.

406

In this connection it is noteworthy that in the severely collapsed
cases with subnormal temperatures spironemata were not discovered
in thick films until reaction had set in and the temperature mounted
to 100°F. or more. Owing to the system adopted of taking the
temperatures of all contacts and suspects and of carrying out a
routine blood examination of every person segregated, whether he
suffered from pyrexia or not, conclusive evidence was obtained as
to the absence of the organism in the blood in the apyrexial state
with the sole exception of the case described above. Spironemata
were not found in the specimens of sputa and urine obtained from
relapsing fever patients.

ANIMAL EXPERIMENTS

The following animals were inoculated with blood obtained from
cases of relapsing fever at the Colonial Hospital or at the Contagious
Diseases Hospital, Labadi:—White rats, black rats, Cricetomys
gambianus, guinea-pigs, monkeys and one rabbit. The quantity
of blood inoculated varied usually from 0°5 ccm. to 2 ccm., citrated
blood being employed in all but one of the experiments. The rabbit
and guinea-pigs proved refractory, no spironemata being at any
time found in their blood, which was examined daily for a fortnight
after inoculation. Eight white rats were inoculated at different
times with infected blood, but in only one of them were spironemata
seen; this rat was given 2 ccm. of blood from a human case on the
21st March and on the following day, twenty-six hours after the
inoculation, two spironemata were found in a thin film of its blood; on
no other occasion in this rat were spironemata found, although the
blood was examined daily for a fortnight. That the blood employed
in the cases of two of these white rats inoculated on the 28th March
was infective was proved by the inoculator unwittingly infecting
himself and developing relapsing fever, the first symptoms of which
appeared on the 4th April—seven days after infection occurred.

Three monkeys were inoculated. Monkey No. 1 received a few
drops of serum only, as the blood, obtained from the first case
diagnosed, was carelessly allowed to clot in the syringe; this monkey
never showed any symptoms and spironemata were never detected

in its blood, which was examined daily for a fortnight after
inoculation.

407

Monkey No. 2—a small baboon—was given 2 ccm. of citrated
blood containing spironemata on the 21st March. On the 23rd it
was not so lively as usual; on the 24th it had a temperature of
102° F’. and spironemata were numerous in its blood; they were less
on the 25th, and had disappeared completely on the 26th. From
this day onwards to the 6th April, when the daily examination of the
blood was discontinued, no spironemata were found. This monkey,
which has been under close observation for three months, has never
shown symptomis of a relapse.

Monkey No. 3—a sooty mangabey—was inoculated with about
2 ccm. of blood containing spironemata on the 23rd April. It
appeared rather subdued on the 25th, but otherwise showed no
symptoms of being ill, and its temperature was only 100° F.; on the
26th spironemata were numerous in its blood, but had disappeared
on the 27th, and after this date no spironemata were found in its
blood. This monkey has also been under close observation for nearly
three months and has shown no signs of relapse.

Three black rats (AZ. vattus) were inoculated. Two received about
1°5 ccm. of citrated blood on the 28th March; this was the same
sample of blood which failed to infect two white rats but proved
infective in the case of the inoculator. Neither of these black rats
showed spironemata in its blood, which was examined daily for
twelve days following the inoculation. A third black rat was given
4 ccm. of citrated blood from a human case on the 2nd July.
Spironemata were fairly numerous in its blood on the 4th, but were
absent on the 5th, and have never been found since that date.

Two Cricetomys gambianus Waterhouse were given large doses
(4 ccm.) of citrated blood which contained spironemata on the same
occasion as the black rat last mentioned. On the 4th July spirone-
mata were found in the blood of one of them; on the 5th both
showed spironemata in large numbers in thick blood films; on the
6th the blood of the first rat which showed spironemata on the 4th
July was free from them, while that of the other showed them in large
numbers; subinoculations were made from each of these rats into
another rat of the same species on this date. Both subinoculated
rats showed spironemata, but at different dates after inoculation;
the rat receiving blood containing spironemata showed them in its
blood on the third day, the rat receiving blood which was apparently

408

free from spironemata on the eighth day. The original two rats
relapsed, one after its blood had been negative for four days, the
other after its blood had been negative for seven days.

A white rat was subinoculated with blood from Monkey No. 3
when it contained numerous spironemata on the 26th April, with a
view to finding if passage through a monkey exalted the virulence
of the strain for white rats. The blood of this rat never showed
spironemata on any occasion, though examined daily for a fortnight
after inoculation.

The results obtained from these inoculations appear to correspond
closely with the inoculation experiments conducted by Gambier
(1923) at Bamako. Gambier found monkeys to be readily infected
with spironemata, white mice to be infected with difficulty, and
rabbits and guinea-pigs to be refractory.

Cricetomys gambianus—the pouched rat—showed itself to be
much more susceptible to infection with spironemata than any of
the other animals employed; it was the only animal which appeared
to relapse. It should be possible to convey the strain to Europe by
means of a series of these rats provided they can be got to survive
the rigours of a northern climate.

CLINICAL MANIFESTATIONS

The first case recorded was I.A., an Italian contractor,
aged 26. He was admitted to hospital on 15th March, 1923, with
pyrexia, headache and severe prostration. He had been resident
in the Gold Coast Colony two years, engaged in contracting work,
and had enjoyed good health. He was taken ill on March 14th.
His blood, on admission, was found free from parasites and pigment.
It was examined again on March 17th, when it was found to contain
numerous spironemata. On March 18th he was given 03 gm.
Novarsenobillon intravenously. The following day, being the sixth
day of the disease, the blood was found free from spironemata, and
the temperature dropped to normal, where it remained until the
patient’s discharge from hospital on March 24th. There was no
relapse, and six weeks later I. A. sailed for Italy, apparently in
perfect health. The other symptoms in the case were icterus of the
seclerae, severe pains, especially in the thighs, vomiting, enlargement

409

of the spleen, and a severe nephritis. The urine contained albumen
and a heavy deposit of granular casts; all this cleared up by
March 23rd.

The only other European case was accidentally infected on two
occasions. His illness ran a similar course, excepting that jaundice
and nephritis did not occur. In this case the rheumatic pains,
especially in the thighs, were excruciating. The second attack was
quite similar to the first, and occurred after an apyrexial period of
three months. It was much milder in character. Both attacks were
treated with the intravenous injections of 0.3 gm. Novarsenobillon,
and there were no relapses.

I. A., Italian, Male, Age 26. Relapsing Fever. Admitted 15th March.
March, 1923

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The clinical manifestation in natives presented a much more
varied picture.

Owing to the necessity for the employment of one and sometimes
two interpreters in the case of Northern Territory patients, it was
far from being an easy task to obtain reliable information regarding
symptoms.

The incubation period was not definitely established, but
appeared to vary from seven days in the case of an accidental
infection resulting from spironemata in a drop of blood from a

410

patient, entering the system by way of a bruised nail bed, to twelve
days in the case of another volunteer who allowed himself to be
bitten by infected lice, and who is thought to have scratched a louse
into his skin. In the case of two other volunteers, infection took
place within eight or nine days of their receiving an emulsion of
crushed lice rubbed into the scarified skin.

Prodromal symptoms were rare. The onset of the disease
appeared to be sudden, and was often accompanied by a distinct
rigor. Frontal headache became so marked in some cases as to
warrant description by the patient as a pain like ‘hammering on
the temples.’

Some of the patients complained of severe pains in the cervical
and lumbar regions, in thighs, shins and wrists. Prolonged attacks
of shivering, rendering the taking of temperatures quite impossible,
and very profuse sweating, were noticed in certain cases. Vomiting
occurred in all but the mild cases, and persisted in some for two to
three days after the fall in temperature, whether preceded by
intravenous medication or not. The vomiting was at times bilious,
but usually occurred after a drink of Akasa—a kind of pap—or
after taking other food. The tongue was coated with white fur, and
anorexia was marked during the course of the disease but gave place
to a ravenous appetite in the majority of those who received an
intravenous injection of Novarsenobillon. Thirst was severe,
particularly, as would be expected, in those cases which suffered
from frequent attacks of vomiting. The patients complained of
giddiness when they attempted to stand; in some the gait was
staggering, and others not only felt too giddy to stand in an erect
posture, but collapsed when they even attempted to sit up. The
asthenic condition persisted in some after convalescence had been
established.

Both liver and spleen were enlarged in many cases, but as the
patients for the most part came from malarial infected areas, it is
possible that malaria was the cause of the splenomegaly, although
diminution in size of the spleen was recorded during convalescence
(quinine not being administered). Jaundice was present in remark-
ably few cases. In one series of one hundred and seventeen, only
three patients suffered from jaundice.

The majority of the patients suffered from constipation, but a

411

certain number, particularly those in whom the temperature had
fallen by crisis, suffered from diarrhoea.

The urine showed albuminuria during the pyrexial stages of the
disease, but was not remarkable for any peculiarities. Oedema of
face and hands, suggesting a nephritis, was seen in a small minority
of cases.

Cough and a small degree of bronchitis were present in many of
the cases, and in one pulmonary signs were so marked as to lead to
a provisional diagnosis of pneumonia being made. Except in this
case the pulse-respiration ratio was normal.

Mental symptoms were observed in a certain number of cases,
and varied from a slight vacuity of mind and loss of memory to
profound mental dulness and, in a small number, to active delirium
and a comatose condition.

The pulse rates recorded were for the most part consistent with
the height of the temperature in the pyrexial period, although in three
cases out of a series of one hundred and seventeen in whom conva-
lescence was prolonged owing to cardiac dilation, the rate remained
unduly rapid for two weeks or more after the fall of temperature to
normal. In the collapsed cases the pulse was of poor volume, thin
and thready and often uncountable.

A somewhat unusual series of facts were noted in connection with
the temperature recorded. It might have been assumed with all
fairness that the severity of other signs and symptoms in a case of
spirillar fever was proportionate to the height of the temperature.
This was by no means the case. In some cases where the tempera-
ture rose to 104° F’. or higher the symptoms were by no means severe,
and recovery rapidly took place after suitable treatment. In other
cases where the temperature did not rise much higher than 100° F.
other signs and symptoms were grave. In the single case under the
care of one of us (P.S.S-C.) which ended fatally the temperature on
admission was 99°F. A thick blood smear was taken and large
numbers of spironemata were observed. The patient was given an
intravenous injection of 0°6 gm. Novarsenobillon and then put to bed.
His temperature rose to 101° F. by 6 p.m. the same evening. On the
following morning his temperature had fallen to g9°F., but his
condition was grave and he could not be persuaded to take any
fluid nourishment. A blood smear was negative to spironemata.

412

He vomited bilious-looking material twice during the day, and by
6 p.m. his temperature was 994° F. When seen on the following
morning at 6-30 a.m.—probably the fourth day of his illness—he
was found to be comatose. The thermometer did not register any
temperature and his radial pulse was so small in volume and rapid
in frequency as to be almost imperceptible and quite uncountable.
A blood smear was negative to spironemata. Efforts were made to
combat the condition by raising the end of his bed, by applying
hot blankets, hot water bottles and by administering intravenous
and subcutaneous salines and brandy. By 10-30 a.m. his tempera-
ture had risen to 972° F. and his pulse, though rapid (120), was of
good volume and tension. Respirations which had been of the
Cheyne-Stokes variety at 6-30 a.m. were now comparatively normal
although the breathing was stertorous. By 2-30 p.m. the patient’s
temperature had risen to to1° F. His pulse was of good volume and
tension and about 120. Breathing had become markedly stertorous
and respirations numbered 36 to the minute. A blood smear was
taken but no organisms were found. The patient died at 4 p.m. on
the same day, having remained unconscious for over 48 hours. As
far as could be gathered, the patient had been ill for two days prior
to his admission to hospital, thus death took place within five days
of the initial symptoms. The above case has been given at some
length in order to show that the height of the temperature had little
relation to the severity of an attack. It is to be noted that the blood
smears taken on the second and third days of the illness prior to
the patient receiving Novarsenobillon showed a very heavy infection
of spironemata.

Another type of temperature was seen in the case of a female
subject of good physique and aged 20. The temperature recorded
on the first four days of her illness was 100° F. or less. A blood
smear taken on the first day proved on examination to show a
moderate number of spironemata. On the second day—the patient
remaining without treatment—the number of organisms in a thick
film was very small, and on the third day none were found. By the
fifth day the temperature had fallen to 97°4° F., and it remained low
for ten days. On the tenth day following the initial fall to below
normal, the temperature rose to 100° F. On the following morning
at 6-30 a.m. the temperature was 104'4°F. Spironemata were not

413

found in the blood until the eleventh day following the original
commencement of the apyrexial period after the first attack. Owing
to the obvious suffering of the patient and to her serious condition,
one of us (P.S.S-C.) did not feel justified in withholding Novar-
senobillon any longer and gave 0'6 gm. intravenously at 11 a.m. By
6-30 p.m. the temperature had fallen to 103° F. On the following
inorning the temperature still stood at 100° F. but fell to 99° F. the
same evening and to 97° F-. by the next morning. Spironemata were
found in moderate numbers at the height of the relapse but not
subsequent to the treatment with Novarsenobillon.

In a second patient—an adult male aged 44—untreated until the
first relapse, the temperature on admission on the third day of his
illness was 103.2°F. Spironemata were present in moderate numbers
ina thick blood smear. The patient received no treatment other than
a cold sponging, and his temperature fell the day after his admission
to 97° F., remained normal or subnormal for two days and then rose
to 101° F, During the apyrexial period, organisms were absent from
blood smears, but were present on the day of the relapse. The
patient appeared to be suffering considerably during the relapse,
and it was not considered fair to him to withhold specific treatment
any longer. [he temperature fell to normal and spironemata
disappeared from blood smears within twenty-four hours of the
patient being injected with 0°6 gm. Novarsenobillon and no further
relapse occurred.

Particulars of a fourth case are worthy of record since the patient
appeared to be suffering from pneumonia on admission. ‘The
patient was admitted to hospital on the second day of the disease.
His temperature, pulse and respiration at 2-45 p.m. on the day of
admission were respectively 103°6° F., 100, and 40. Although a well
nourished male of 25 he was too weak to move hand or foot and was
delirious. Bronchitic rales were heard over both sides of his chest
and signs of early pneumonic consolidation were heard over the left
lower lobe. A blood smear showed the presence of spironemata in
large numbers. The patient was very jaundiced. He was given
0°6 gm. of Novarsenobillon intravenously and his temperature, which
‘rose to 104°F. by 6 p.m. the same evening, fell to 100° F. on the
following day and then to 97°F. on the morning of the third day
after his admission. Subsequently, the temperature rose again on

414

the evening of the third day following admission to 998° F. and
to 100° F. on the morning of the fourth day, but fell to normal on
the same day. From thence onwards the temperature went to a few
points above normal for the next fourteen days and then steadied
down to subnormal. The pneumonic signs cleared up without any
signs of resolution, but the patient suffered from bronchitis for a
fortnight following his admission to hospital. Spironemata were not
found in the patient’s blood subsequent to the treatment with
Novarsenobillon.

When discussing the variations in temperature in cases of
relapsing fever it would be unwise to take the four cases quoted above
as typical examples. By far the majority of cases suffered from
temperature varying from 100° F. to 105° F., though a small number
showed spironemata in blood smears with a temperature of only
99° F. In most cases a fall of temperature to normal or subnormal
took place within twelve hours of treatment with Novarsenobillon,
although in some cases the temperature remained above normal
though lower for two to three days and then fell. During the early
days of the epidemic when only 03 gm. of Novarsenobillon was
administered, a number of the patients relapsed after varying
intervals. In a small minority of cases it was found necessary to
give o'6 gm. of the drug followed by 03 gm. after an interval of
three days.

RELAPSE

The information regarding the occurrence of relapses is scanty
for two reasons. Africans, and in this they resemble all races, do
not take kindly to hospital treatment, and purely medical cases
prefer to remain in their own homes rather than to enter hospital,
however comfortless the former may be, and however much their
chances of recovery may be so impaired. It follows naturally that if,
owing to pressure having been brought to bear on them, they have
been admitted to hospital for treatment, their one aim and object
is to obtain their discharge therefrom as soon as possible. Conse-
quently, when a case of relapsing fever was admitted to hospital,
endeavours were made to sterilise the patient as regards the infecting
organisms in his blood and to effect his cure with the least possible
delay. By making his stay in hospital as short as was compatible

415

with his own well-being and with the safety of the general public,
other cases occurring in the town were encouraged to seek medical
attention as soon as they became infected, instead of remaining con-
cealed from the health authorities and so helping to spread infection.

Secondly, it was not considered justifiable, except in a very
small number of cases, to withhold treatment from a patient with a
view to determining the approximate length of the apyrexial period
between attacks, since by so doing, the well-being and possibly even
the life of the patient was placed in jeopardy.

The relapses observed can be divided into two classes, one in
which the patient had received no specific treatment, but only general
symptomatic treatment as, for example, light diet, saline purge, cold
sponging, and the second class in which specific treatment with
Novarsenobillon had been administered.

Examples of the first were afforded by the cases of a man and
woman. The man was admitted on the third day of his illness
suffering from the usual symptoms of relapsing fever and with a
temperature of 103'2°F. Spironemata were present in moderate
numbers in a thick blood film. The patient received symptomatic
treatment only and his temperature fell to 97° F. on the day following
admission and remained normal or subnormal for two days. On the
third day following the crisis which terminated the first attack, the
temperature rose to 101°F., and spironemata which had been absent
from blood films during the apyrexial period of two days were again
found in blood films. The condition of the patient did not justify
the withholding of treatment further. The woman suffered severely
from headache, pains in the back, legs and wrists, but her tempera-
ture during the first four days of her illness did not exceed 100° F.,
although a blood film taken on the first day showed a moderate
infection with spironemata. The number of spironemata seen on the
second day of the disease was smaller, while none were found on the
third and fourth days. On the fifth day of the disease the tempera-
ture fell to 944° F. and remained low for ten days. On the tenth
day following the initial fall to below normal the temperature rose
to 100° F. and on the following morning at 6-30 a.m. to 104°4°F.
Spironemata were found in moderate numbers, but the patient’s
condition was sufficiently serious to make further delay in the
administration of specific intravenous medication quite unjustifiable.

416

Several examples of the second series of relapses, that is to say,
relapses occurring in spite of treatment, occurred amongst the cases

Arman, Female, Age 20. Relapsing Fever. Admitted May 18th.
May, 1923 June

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fed

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BBBBBBEEE EEE Cann ‘
Paes HNC ROESED a SAS EVES AEE 9h
BBSBUE ESSE ene A BEBE 2
| Bet EES ER Dy EPR EY VEY Ea
SES Ese ET VET EAE LETT SE DSS
TANCE CE Nad
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HAnBBaAnEadnaHEnnaon
Em

S
ey

arasites

Serpu Syrz, Male, Age 44. Relapsing Fever. Admitted June rst.

June, 1923

=
Pe pes

‘e a > ‘. ‘e e ° o

treated in the Contagious Diseases Hospital. The following case is
remarkable for its severity and for the unduly long period that
elapsed between the patient’s apparent cure and his relapse. The

417

patient, a well nourished young adult of 23, was admitted to
hospital together with a large number of other cases, suspects and
contacts, on 29th April, 1923, suffering from signs and symptoms
of relapsing fever. On the 30th April his temperature was 105° F.

Mumunt III, Male, Age 23. Relapsing Fever. Admitted May 2nd.

May, 1923 First attack.

F269 ESOS Ee

lapels shee eer etl
REE EER REECE EG ET

Chart to illustrate occurrence of relapse after an unusually prolonged period of apyrexia possibly
resulting from the administration of an inadequate dose of Novarsenobillon during the first attack.

Mumuni III, Male, Age 23. Relapsing Fever. Admitted May 26th.

Relapse.
May, 1923 June

: ata Hee Rite Hees
or CA a TA
ef tt MAM TT et ti ded

srasites Fe et ea

418

On the following morning his temperature was 102°6° F. but a blood
film failed to show the presence of spironemata. His temperature
on the following day was 105° F., spironemata were present in a
blood film, and he was given 0°3 gm. Novarsenobillon intravenously.
By the evening of the same day the patient’s temperature had fallen
to 103° F. The next morning the temperature had still further fallen
to gg'6° F., and by the evening to 98°4° F. For the 3rd to the oth
of May inclusive the temperatures were 96°, 99°, 98°4°, 98°6°, 99'2° F .,
but no spironemata could be found in blood films taken during this
period. ‘Thereafter the temperature remained normal or subnormal
until the 21st of May when the patient was discharged. Three days
later on the 24th of May (fourteen days after the patient’s temperature
had fallen to normal) the patient complained of anorexia, diarrhoea
and frontal headache. He was seen on the 26th of May and found
to have a temperature of 100°F., and spironemata were observed in
his blood film. On this occasion he was given 0°6 gm. of Novar-
senobillon. Huis temperature rose to 102° F. by 6 p.m. the same
night, but had fallen to 99° F. at 6 a.m. on the following morning.
Organisms, however, were still visible in blood films. The tempera-
ture fell to 97'4° F. the same evening and subsequently remained
normal or subnormal until the 13th June when he was discharged,
his temperature not having been raised for sixteen days. The
interesting point about this case is that the original pyrexial period
lasted for about eleven days, followed by an apyrexial period of
about fourteen days, when a relapse occurred. ‘The pyrexial perioa
during the relapse lasted for four days and was followed by a period
of apyrexia for sixteen days. The explanation in this case is that
the o°3 gm. of Novarsenobillon administered was too small a dose to
sterilise, but that it probably had the effect of retarding the relapse
which took place fourteen days after the patient’s temperature had
been normal or subnormal. During the relapse, spironemata were
present in large numbers in blood films until the third day of the
disease, when a 0'6 em. dose Novarsenobillon was given. Organisms
were present in small numbers on the morning following the
injection, but disappeared from blood films taken thereafter.
Several similar cases occurred in which an initial dose of 0°3 gm. of
Novarsenobillon appeared to effect a cure, but in which it ultimately
proved to be insufficient in preventing the occurrence of a relapse.

419

A few patients relapsed even after receiving 06 gm. of the drug,
but for routine work, dealing with a large group of persons, this
dose appeared to be satisfactory, followed by 0°3 gm. or 06 gm. in
the small number of cases failing to react to the initial dose, or
showing signs of relapse.

Briefly, in untreated cases relapses occurred with an apyrexial
period varying from two to ten days, while in treated or partially
treated cases the apyrexial period varied from two to fourteen days.
As a rule, but not invariably, the relapse was less severe both in
signs and symptoms and also in duration than the initial attack.
Out of the hundred and seventeen cases that came under the care
of one of us (P. S.S-C.) twenty-three, or 19 per cent., relapsed on
one occasion, and eight, or 6°8 per cent., relapsed a second time.
Thus the total number of relapses in the one hundred and seventeen
patients was thirty-one, or 26°5 per cent., of all the patients.

The following table shows the results :—-

Tasre IV.
Blood film
Positive Negative Unknown Total
First Relapse... Fac oa II 7 5 23
Second Relapse ... te “o 3 5 ete 8
Totalg~*..: ode Fd 14 12 5 31

In cases where the organisms could not be found in blood films,
the diagnosis of relapse was based upon rise of temperature and the
recurrence of the signs and symptoms of the original attack.

IMMUNITY

Second Attacks. Immunity 1s said to be of short duration in
relapsing fever. It would be unfair to draw any such conclusions
from the epidemic under review, since so little time has elapsed
since the occurrence of the cases described above. One undoubted
second attack occurred, however. The patient, a European, originally
became accidentally infected with relapsing fever while injecting

420

infected blood into a rat. Seven days later he developed the signs
and symptoms of the disease. At first no spironemata were seen in
his blood, but after two days of moderately severe pyrexia the
organism was found to be present in small numbers. The patient
was treated with 0°3 gm. Novarsenobillon intravenously, and rapidly
recovered. This first attack occurred in the beginning of April.
At the end of June the same individual contracted a severe attack
of the disease as the result of feeding infected lice on his forearms—
two escaped and are thought to have been scratched into his skin.
The second attack was rather less severe than the first, but reacted
to treatment with Novarsenobillon as rapidly as had been the case in
the first attack. If the patient acquired any immunity from the first
attack, and this is probable, since he subsequently carried out a
series of experiments, feeding on his forearms lice from relapsing
fever patients, the immunity was of decidedly short duration, in
fact less than eleven weeks.

TREATMENT

Apart from intravenous medication with Novarsenobillon given
intravenously in 10 c.c. of warm, sterile, distilled water in doses
varying from o°3 gm. in the early cases to 0°6 and o'9 or I1'2 gm.
(divided into two to three doses) in the later cases, the treatment
administered to patients was symptomatic. Patients were washed,
shaved and disinfested on admission, and put to bed with a sleeping
mat and two warm blankets. They received their iniection of
Novarsenobillon as far as possible on a fasting stomach. If they
wished, they were given two biscuits and a drink of milk a short
time after the injection. Cold sponging was resorted to where the
temperature was 103° F. or higher, but otherwise, apart from being
given as much water in small quantities as they wished, the patients
were left to sleep quietly until the following morning, care being
taken to avoid chills in cases where the temperature fell by crisis. The
following morning a saline purge was administered, and if conditions
were satisfactory and no vomiting was present a light diet was given.
It was noticed that the patients not only wanted to get up and
resume their usual everyday life immediately after the fall of
temperature had occurred, but that they wished to resume a normal
diet at once. As a rule the attack left the patients somewhat weak,

421

and vomiting immediately occurred, if the patients were allowed
to satisfy their ravenous hunger. In those patients who reacted well
to the treatment and in whom no elevation of temperature occurred
within the fourteen days subsequent to the fall of temperature to
normal, attempts were made as far as possible to graduate both diet
and exercise until, for some days before the patients were due for
discharge, they had resumed a normal hfe.

In order to obviate the possibility of relapses, patients were kept
in hospital until their temperatures had been normal for fourteen
days. Blood smears were taken of all patients with temperatures

Buxart, Male, Age 38. Relapsing Fever. Admitted July 2nd.

July, 1923

ae SE ate eT A a a ae

Typical moderately severe case showing fall of temperature by crisis 24 hours after administration
of o°6 gm. Novarsenobillon.

above 99° F., and even if spironemata were not found in the blood
films and other conditions could be excluded, further treatment with
Novarsenobillon was carried out. This step appeared to be
justified in the light of the subsequent histories of such cases.
Among the one hundred and seventeen patients admitted to the
Contagious Diseases Hospital, eighteen had already, received an
intravenous injection of 0°6 gm. Novarsenobillon. Injections—three
of which were given intramuscularly owing to the inability to
discover a vein of adequate size—to the number of one hundred and

422

thirty were given by one of us (P.S.S-C.). Of the total number
of injections one hundred were of 0°3 gm. and forty-eight of 06 gm.
Patients who received but one injection numbered one hundred and
seventeen, those who received two numbered twenty-three, while
eight patients had to receive a third injection. Generally speaking,
apart from a little vomiting for two or three days after the injection
(this vomiting occurred in untreated cases and consequently may
have borne no relation to the intravenous medication) no ill-effects
were experienced from the use of Novarsenobillon. One death occurred
in a series of one hundred and seventeen cases that came under the
notice of one of us (P.S.S-C.), but the patient was in a serious
condition prior to the injection, and when death occurred on the
third day following the injection, it could be attributed with fairness
to the disease. West Africans appear to tolerate organic arsenical
preparations exceedingly well.

PREVENTIVE MEASURES

The control of the outbreak of relapsing fever in Accra during
the earlier stages was hampered by two considerations, viz.,
ignorance of the vector and mode of transmission of the particular
strain of organism and lack of legal powers to deal with cases,
suspects and contacts. The vector being unknown, lice, ticks,
bed bugs, mosquitoes and biting flies were all treated as suspect.
Samples of all these were collected either, as in the case of the first
two from patients known to be suffering from the disease or, as in
the case of the remainder, from bedding, mats and other articles in
infected premises, and were taken to the Medical Research Institute.

Measures were taken against all possible vectors, and with this
end in view attention was concentrated on premises situated in
congested areas of the town occupied by Hausas and members of
Northern Territory tribes, whose habits in regard to overcrowding
and a marked aversion from cleanliness, adequate lighting and
ventilation were well known. Careful house-to-house visits were
made in those areas and throughout the town, as many as 46,013
being carried out during March, April, May and June, one of us
(P.S.S-C.) being responsible for 1,006. During these inspections
personal, domestic and general cleanliness was preached, all old

423

sacking, lousy bedding and other refuse being removed from
compounds. Moreover, efforts were made to see that all houses
were provided with adequate lighting and ventilation. During
these visits a practice was made of urging any person who appeared
to be ill and suffering from fever to go for treatment to the Native
Hospital.

The procedure adopted early in the epidemic when a case of
relapsing fever was reported was for the patient to be admitted into
the Native Hospital, for his quarters to be disinfected and disinfested
and for a watch to be kept on all contacts, any of whom showing signs
of fever being urged to report to the Medical Officer at the Native
Hospital.

By the fourth week of the outbreak it was evident that infection
which appeared to have been introduced into Accra from bush
villages was spreading to other parts of the town from houses already
infected with relapsing fever. To obviate this tendency to spread,
it was strongly urged that legislation should be passed, in order that
the Health Authorities might round-up all persons suffering or
suspected to be suffering from relapsing fever, together with
contacts with such cases, for the purpose of segregating them and
sterilising them as far as concerned the presence of spironemata in
their blood. Legislation was not passed, however, until the 28th of
April, by which time the disease had appeared in several parts of
the town, though principally in Tudu and the Zongo. On the
29th of April, acting within the powers obtained through this legisla-
tion, it was possible to effect a round-up of cases, suspects and
contacts on a much larger scale, so that within twenty-four hours of
legislation being passed, over three hundred and _ sixty-three
patients, suspects and contacts were removed from infected premises
to the Contagious Diseases Hospital. Where necessary, one or two
contacts were allowed to remain in such infected premises, to
safeguard the property from thieves, to keep the premises clean and
to care for any horses or cther animals. Such persons were visited
daily to exclude the possibility of their having contracted the
disease.

The health of the person permitting, a routine was adopted
at the Contagious Diseases Hospital in almost every case. On
admission all male cases, suspects and contacts had their heads,

424

armpits, beards and pubic hair shaved, while in the case of females
the hair was close clipped and shaved from their armpits and pubes
under the supervision of a female sanitary inspector. Subsequently
the shaving was foliowed, where physical conditions permitted, by
a sea bath and by a wash-down where a sea bath was inadvisable.
All clothing was shed into barrels containing a 5 per cent. solution
of Izal prior to the bath being taken, and after the bath a warm
blanket, sleeping mat, and cup and plates were issued to everyone.
Temperatures and blood smears were then taken and recorded and
the groups dealt with were allotted accommodation in three classes
of huts, according as to whether they were thought to be suffering
from relapsing fever or were merely suspects, or contacts with cases
and suspects. Special diet, as for example, milk, tea, broths, etc.,
was given to patients, whilst the remainder received two meals per
day. Hot Akasa was given in the early morning as soon as
temperatures had been taken, and was followed at mid-day by a
large meal of rice, plantain, fula or other foodstuffs purchased in
the markets. As far as possible the tastes and wishes of patients
and contacts were consulted with regard to the variety of food
supplied. Contacts were detained for fourteen days, during which
time they were given a certain amount of work to do in the way of
scrubbing out huts, keeping the segregation compounds clean,
helping with the chopping-up of firewood, with the preparation of
food and drawing of water. They enjoyed sea baths daily,
arrangements being made for the opposite sexes to bathe at different
times. At the end of the quarantine period, if their temperatures
which were taken morning and evening had remained normal, the
contacts were again submitted to a thorough shaving, and were
given a bath, and then had their disinfected and disinfested clothing
returned to them and thereafter were discharged. Contacts or
suspects who developed raised temperatures, or in whom blood films
proved to be positive as regards the presence of spironemata, were
immediately transferred to the huts reserved for patients. After
receiving appropriate medication, patients were kept in hospital until
they had been free from pyrexia for fourteen days—blood films being
taken daily while temperatures were raised.

It is noteworthy that the following method of disinfecting and
disinfesting clothes and blankets appeared to give the best results.

425

The articles to be disinfested were first soaked for forty-eight hours
in barrels containing 5 per cent. solution of Izal. They were then
washed and placed in the sun during the middle of the day on
sheets of corrugated iron. ‘This resulted in most efficient disinfesta-
tion, for the heat generated was at least 150° F. Neither lice nor
eggs capable of hatching survived this treatment. Purses and
amulets, the latter carried in great numbers by Hausas and Northern
Territory tribesmen, required special treatment.

In order to minimise the risk of infection being carried from the
hospital to Accra, the auxiliary staff of the hospital were persuaded
to stay in special quarters reserved for them in the grounds of the
hospital, and all the staff, including one of us (P. S.S-C.), took
further precautions by frequent baths and by shaving the hair from
axillae and pubes. A police guard was maintained at the hospital
during the period of the outbreak, and the Non-Commissioned
Officer in charge is to be congratulated in not losing a single patient
or contact.

CONCLUSIONS

1. This first recorded outbreak of relapsing fever in British
West Africa is due to a spironema conveyed by lice.

2. As regards inoculation experiments, monkeys, black and
white rats become infected with the strain, but do not relapse ;
guinea-pigs and rabbits are refractory, whilst the pouched rat
becomes infected and relapses.

3. The vectors of the organism in the present epidemic and
the inoculation experiments suggest that the parasite is not the
Sp. duttoni, but corresponds more closely to Sp. recurrentis (vel
obermeiert), or a related strain.

4. Novarsenobillon is a specific in the treatment of the disease.

5. Immunity does not appear to be lasting or complete in cases
treated with Novarsenobillon.

REFERENCES

Batrour, A. (1922). Official History of the War; Med. Serv., Diseases of the War, Vol. I,
Chaps. XIII and XIV.

Batrour, A., and Bousrietp (1911). Fourth Report of the Wellcome Tropical Research
Laboratories, p. 62.

426

Brent, A. (1908). On the Morphology and Life History of Sp. duttont. Anns. of Trop. Med.
and Parasitol., Vol. I, p. 435.

Brumpt, E. (1922). Les Spirochétoses. Nouveau Traité de Médecine. No. 4, p. 491
(quoted in Rev. App. Ent., Ser. B, Vol. XI, p. 30).

Cracc, F. W. (1922). Relapsing Fever in the United Provinces of Agra and Oudh. Jnd.
Four. Med. Res., Vol. X, p. 78.

Gambier, A. (1923). Essai de transmission aux animaux de laboratoire du spirochéte de la
fiévre récurrente soudanalse Bull. Soc. Path. Exot., Vol. XVI, 159.

Kerrest, J., Gambier, A., et Bouron, A. (1922.) La fiévre récurrente au Soudan. Bull. Soc.
Path. -Exot., Vol. XV, p. 320.

Macriz, J. W., and Yorke, W. (1917). The Relapsing Fever Spirochaetes. Anns of Trop.
Med. and Parasitology, Vol. X, p. 81.

Manson, J. K., and THornton, L. D. H. (1919). East African Relapsing Fever. ‘Four. of
R.A.M.C., Vol. XXXIII, pp. 97 and 193.

Nicoxte, Cu., Biaziot, L., et Consett, E. (1912). Etiologie de la fiévre récurrente. Sa mode
de transmission par le pou. C. R. Acad. Sci., Vol. CLIV, p. 1636 (quoted by Edm.
Sergent et H. Foley).

Nutta.t, G. H. F. (1917). Parasitology, Vol. X, p. 107.

SERGENT, EpM., et Forzy, H. (1922). L’Epidémiologie de la fiévre récurrente dans I’ Afrique
du Nord. Trans. Roy. Soc. Trop. Med. and Hygiene, Vol. XVI, p. 170.

427

CARBON TETRACHLORIDE IN
FILARIASIS

BY
Deer LE Hie

(Sir Alfred Lewis Jones Research Laboratory)
(Received for publication 23 August, 1923)

In view of the successful treatment of intestinal nematodes with
carbon tetrachloride administered orally, it was decided to try the
effect of the drug administered intravenously and intramuscularly
on filariasis.

Prior to treating infected human beings, a series of experiments
were carried out on healthy dogs. It was found that dogs could
stand relatively large amounts of the pure drug administered
intramuscularly or intraperitoneally, and that older animals tolerated
the drug better than young animals; the minimal lethal dose was
found to be 0°25 ccs. (2.é€., 0°39 gms.) per kilo body weight in
animals weighing less than 1 kilo, whereas animals weighing over
4 kilos showed symptoms from which they recovered after a dose
of 0°6 ccs. (2.e., 0°94 gms.) per kilo body weight; these symptoms
were drowsiness and refusal to take food for a few days after the
injection.

Injected intravenously the drug caused rapid death due to
embolism, but when mixed with two parts of ether by volume,
embolism after intravenous injection was avoided.

An intramuscular injection of 0'5 ccs. of the drug into a healthy
human being caused irritation at the site of injection, which was not
severe and passed away in a few minutes; shortly after the injection
the distinctive taste of the drug was felt in the mouth.

After these preliminary experiments the drug was tried on four
adult patients each with a slight infection of Hlarza bancrojti, as
judged from the number of microfilaria in the circulating blood.
In each case the number of microfilaria per c.c. was estimated by
counting the number of microfilaria in 20 cmms. of blood at 9 p.m.,
before commencing treatment and at the end of treatment.

428

Case 1. A native aged 35, weight 144 lbs.
Number of microfilariae in the blood at 9 p.m. before commencement of

treatment—So per C.c.
25.6.23. Intramuscular injection of 0.§ c.c. carbon tetrachloride.

20.7.23. Intravenous injection of I c.c. carbon tetrachloride mixed with

2 scree. chien:
25.7.23. Intramuscular injection of 1.§ c.c. carbon tetrachloride.
30.7.23. Intramuscular injection of 2 c.cs. carbon tetrachloride.

Case 2. A native aged 32, weight 122 lbs.

Number of microfilariae in the blood at 9 p.m. before commencement of

treatment—I0o per c.c.
20.7.23. Intravenous injection of I c.c. carbon tetrachloride mixed with

2.0.08.) ether.
25.7.23. Intramuscular injection of 1.5 c.cs. carbon tetrachloride.

Cast 3. A Native aged 25, weight 143 lbs.

Number of microfilariae in the blood at 9 p.m. before commencement of

treatment—350 per C.c.
11.7.23. Intramuscular injection of I.1 c.cs. of carbon tetrachloride.

16.7.23. Intravenous injection of I c.c. carbon tetrachloride mixed with

2. C.Co welch
25.7.23. Intramuscular injection of 1.5 c.cs. carbon tetrachloride.
30.7.23. Intramuscular injection of 2 c.cs. carbon tetrachloride.

Case 4. A native aged 24, weight 154 lbs.

Number of microfilariae in the blood at 9 p.m. before commencement of

treatment—I0o per c.c.
20.7.23. Intravenous injection of I c.c. carbon tetrachloride mixed with

Ze Comeaner,
25.7.23. Intramuscular injection of 2 c.cs. carbon tetrachloride.
30.7.23. Intramuscular injection of 2 c.cs. carbon tetrachloride.

All the intramuscular injections were given into the buttock.

The patients were observed for one hour after each injection.

The intramuscular injection caused local pain, which passed away
in from five to ten minutes. No marked analgesic effects were
noticed after the local pain had disappeared. Injection of even
2 ccs. of the drug intramuscularly produced no anaesthesia.

Generally there was a slight diminution in the pulse rate up to
four beats per minute following intramuscular injection.

All the cases noticed the taste of the drug shortly after injection.

Intravenous injection of the drug mixed with ether caused a
severe attack of coughing, which commenced during the injection
and lasted a few minutes; one case complained of a burning sensation
in the mouth; all the cases were sleepy after the intravenous injection,

429

but whether the sleepiness was caused by the ether or the carbon
tetrachloride it is impossible to say.

None of the cases showed albuminuria or other ill-effects either
during or after the treatment, which was abandoned after the
30th July, 1923.

None of the cases showed any marked diminution of the micro-
filaria in the blood after the treatment, but it is impossible in the
case of Filaria bancrofti to form an opinion of the effect of the drug
on the adult worms.

The action of intravenous or intramuscular injections of carbon
tetrachloride on adult filaria can only be tested in cases of Loa loa,
but up to the present no suitable cases have been found in Freetown.

In view of the comparative safety with which the drug can be
administered, both intravenously and intramuscularly, it is hoped
that it will be tried on cases of Loa loa in localities where that disease
is common.

Penave to thank’ Dro*]> YsWood;"of-the W.A.M.S., for the
opportunity of carrying out the above treatment, and Dr. P. A.
Maplestone, from whose series of routine examinations for parasites
the cases were selected.

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431

YELLOW FEVER IN THE GOLD
COAST: ITS ENDEMIC AND
EPIDEMIC CHARACTER

Rar Qty WEEE

WEST AFRICAN MEDICAL SERVICE
(Received for publication 24 August, 1923)

The recently reported cases of yellow fever in different parts of
the Gold Coast* bring into prominence again the subject of its
endemicity in West Africa, and suggest a consideration of the
conditions which give rise to these periodic outbreaks.

That the West Coast of Africa has been an endemic centre no one
disputes, but opinion is divided upon the question of the presence
of yellow fever as an endemic disease to-day.

The Yellow Fever Commission (West Africa) appointed by the
Colonial Office in 1913 considered the evidence submitted was in
favour of the belief that West Africa is an endemic centre (1916).
More recently, Guiteras (1921) and Hoffman (1921) expressed the
opinion that this centre has already ceased to exist. This supports
the view held by Ross, who suggests that the Stegomyia mosquito is
not sufficiently numerous to permit the disease to maintain itself.

The investigations of the Rockefeller Foundation in 1920 failed
to discover a single case, and we find that out of nearly half a million
recorded illnesses treated in Nigeria during the years 1919-21, there
is but a single diagnosis of yellow fever.

In spite of this formidable array of opinion and facts, it is not
possible to ignore the significance of the existence of a disease which
does not conform to prevalent types of fever and which is capable of
being diagnosed as yellow fever.

The reports of the recent cases occurring among Europeans on
the Gold Coast leave very little room for doubt as to the accuracy
of the diagnosis. In all these were the black tarry or coffee grounds
vomit, a marked diminution in the quantity of urine—amounting in

*A cable received at the Colonial Office on July 11, 1923, stated that 25 cases of
yellow fever had occurred in the Gold Coast since November 1, 1922—18 in Europeans, all fatal,
and 7 in natives, 2 fatal.

432

some cases to complete anuria—with albuminuria, associated with
symptoms of extreme urgency followed rapidly by death. When
post-mortem examinations were made, fatty degeneration changes
were found in the liver and kidney. These features were common to
all, but haemorrhage from mucous membranes, conjunctival injection,
varying degrees of jaundice, Faget’s sign and high fever are
described. It 1s also noteworthy that, with one exception, the fatal
termination occurred within a week of the onset of the illness.

In the face of such evidence, it must be admitted that yellow fever
is a disease of West Africa. The next important point is—in what.
form does it exist ?

An examination of the trade routes between the West Coast of
Africa and other parts of the world fail to supply us with an external
source of infection. In the 1911 outbreak at Bathurst it was thought
that the occurrence of the disease at this port was connected with the
visit of the s.s. ‘ Akassa.’ Subsequent enquiries, however, failed to
confirm this. In the epidemic under consideration, there 1s not the
shghtest ground for suspecting that the disease was introduced from
without. It seems, then, that we must assume that yellow fever does
exist in West Africa, in some latent form requiring special conditions
for its development.

That special conditions are required may be gathered from the
infrequency with which the disease occurs among Europeans in any
one place, notwithstanding that the essentials for the maintenance
and spread of the disease are always present—the virus, the carrier
and the non-immune.

The explanation which used to be given for the comparative
freedom from yellow fever among Europeans on the West Coast of
Africa was, that the newcomer usually became the subject of a mild
infection which conferred immunity for the remainder of his stay in
the country. But this contention cannot be maintained, in view of
the fact that victims of the disease have succumbed to it after periods
of residence up to thirty years. It is necessary, therefore, to look for
some other explanation.

When an endeavour is being made to discover the aetiological
factor concerned in the outbreak of an infectious disease, it is helpful
to be able to note the conditions under which it has died out in
certain places.

433

Peterson’s (1922) view of the spontaneous elimination of yellow
fever in St. Thomas—which is based upon Carter’s principle of
‘the failure of the human host ’—suggests that the virus gradually
becomes attenuated until it reaches the point of extinction. In other
words, the commerce between the mosquito and the unresponsive
native eradicates the disease, not by the process of conferring general
immunity, but by the death of the virus. The logical inference
which may be drawn from this is, that the West Coast of Africa
would cease to be a yellow fever area as soon as, or shortly after, less
immune individuals were excluded from the country. The European
is, of course, the obvious non-immune. Yet if we study the records
of yellow fever epidemics, we shall find long intervals between
outbreaks in any one place, and that they are traceable to a
non-European source. This is fair presumptive evidence that the
European is an accidental victim rather than an agent in maintaining
the disease.

It may be urged that mild cases occur among Europeans which
escape diagnosis. The mortality rate in the 1910-11 outbreaks
(1913), which show six recoveries in forty cases, 1s against this; more
especially when the reports of these recovered cases are scrutinised.

Of the six, three are included without any details of their illness,
and as such, merely indicate people who were sick at the time.

Of the three remaining, two are so untypical of yellow fever, that
it 1s safe to presume their inclusion was due to the prevalence of the
disease rather than to the character of their physical signs and
symptoms. The last case of this group appears to have responded,
eventually, to energetic treatment with quinine. Albuminuria was
absent throughout the illness.

In the present epidemic in the Gold Coast, the mortality among
the Europeans attacked is 100 per cent. If we exclude the six
doubtful cases referred to above, it would appear that yellow fever
in West Africa is invariably fatal to Europeans. At any rate, it
may be said it shows itself in a way which is not likely to be over-
looked.

To recapitulate: The indigenous native, dissociated from the
presence of newcomers, is incapable of maintaining the virus. The
disease is not overlooked when it occurs among Europeans. The
intervals which elapse between outbreaks among Europeans are proof

434

that the European is not responsible for the fact that yellow fever 1s
an endemic disease in West Africa.

It is obvious then that some other section of the community keeps
the virus alive in a ‘ larval’ state.

It has been said that previous recorded outbreaks are traceable
to a native source. In the Reports of the 1910-11 epidemic (1913)
it will be found that in every case when mention is made of living
conditions, there is the association of close native proximity to the
European attacked. Further it will be found that the native element
is an imported one, either from the confines of the colony itself, or
from a more remote part of West Africa.

Cases numbers 43 and 44 were Kroo-boys. 37 was a Yoruba
lately come to Lagos, where he was taken ill. 29 and 38 also
occurred in Kroo-boys. 28 was a Hausa. 19 and 20 were of the
Mend tribe living in Freetown at the time they were taken ill.

The foregoing comprise all these cases among natives where it
is possible to identify their nationality. The remainder come under
the, not very illuminating, description ‘A native born and bred in
West Africa.’

In the Gold Coast epidemic of this year, the cases at Saltpond
were traceable to a Kroo-boy who died of yellow fever shortly after
his arrival there. Later, eighteen miles away, Cape Coast was
attacked, isolated cases subsequently occurring at Winnebal, Accra,
Keta and Seccondee.

On the Gold Coast we find that West Africans who come to the
Colony pursue one of the following callings :—Trading, soldiering,
mining, as railway and road labourers, or manual work at the
seaports. With the exception of the last-named class, it will be
found that the principle of segregation is conformed with. The
migratory trader, the Hausa, has his Zonga to go to, the soldier
his barracks, while railway and road labourers (usually drawn from
the Northern Territories) have carefully supervised camps.

The porter of the Gold Coast, the Kroo-boy, has no such
provision made for him, and he is to be found, as a rule, domiciled
in the compound of his employer, usually a European. He also has
access to the native quarters of the town, and consorts with other
Kroo boys who are employed by native merchants. There is here,
then, a connecting link between the two classes which have been

435

excluded as not being responsible for maintaining the disease. The
Kroo-boys also represent, numerically, a section of the community
which must be taken into consideration, and they become suspects,
partly because the process of elimination adopted here has left them
unexonerated, and also for the reason that they are known to contract
the disease and to have been responsible for outbreaks among
Europeans.

The question now arises, how is it that these Kroo-boys are more
susceptible to infection than the indigenous native ?

It will be shown later that the immunity to yellow fever with
which all West Africans are endowed is merely relative in degree
and breaks down under certain conditions. It is suggested here
that one such condition is change of environment.

Apparently the native’s degree of immunity is sufficient so long as
he remains in his own country, but becomes impaired when he goes
to live in another part of West Africa. There is support for this
assumption in the fact that West Africans do contract ‘ fever’ when
they leave one part of West Africa to take up residence in another.
This has been recognised by Government Medical Officers, and the
West Africans themselves are aware of it. An opportunity of
observing this phenomenon occurs when a _ native official 1s
transferred to a new station, or is on leave from a Colony of which
he is not a native. That the ‘fever’ mentioned above is often due
to malaria, there is very little doubt. An intensive and fatal case
of this disease was seen at Accra in a Kroo-boy who had been
resident in the Colony for six months. It is known that malaria
does not attain to such severity in the adult indigenous population
of West Africa. The influence of environment is, therefore, a factor
which must be reckoned with when the subject of immunity is being
considered.

From these premises it 1s easy to reconstruct the sequence of
events. The Kroo-boy who migrates to some other part of West
Africa automatically becomes more susceptible to yellow fever. He
contracts the disease in a mild form, unrecognisable as such, and
causing very little, if any, inconvenience. But an impetus has been
given to the virus which, under favourable conditions, ultimately
becomes so enhanced as to give rise to definite illness. The final
stage is reached when living conditions make it possible for the

436

Kroo-boy to pass on this infection to a European. When this
occurs the virus has attained to a very virulent degree of toxicity,
which if unchecked by the wholesale destruction of the mosquito,
will be capable, ultimately, of infecting—-sometimes with fatal
consequences—the indigenous and erstwhile unsusceptible native.
This happened both at Saltpond and Cape Coast during the present
epidemic, and is proof of what has already been said, that the
West African’s immunity is merely a relative one.

It is well known that any break in the chain of essentials which
go to produce a yellow fever infection is sufficient to stop, or at least
interrupt, the process. The lapse of time which takes place between
observed epidemics in West Africa, seems to suggest that the chain
is delicate in its construction and that the process of building up the
virus sufficiently to produce recognisable effects is a long one.
Segregation of Europeans—in as far as it obtains in West Africa—
appears to have the effect of lengthening the process. It has certainly
provided immunity for the segregated, for in no single instance has
a case occurred among them. When it is remembered that in
segregation areas native servants—often Kroo-boys and natives of
the Northern Territories—live in close contact with the European, it
would seem that the slightest precautions are sufficient to prevent
infection. As Carter suggests in his statement of requirements for
the maintenance of a yellow fever infection, the number of mosquitoes
may fall short of what 1s necessary. It probably will be found also,
that non-interference with the mosquitoes, overcrowding and lack of
light and ventilation are necessary. Routine sanitary work probably
intereferes from time to time with one or other of these subsidiary
requirements, and has the effect of delaying the development of the
virus. But sooner or later, it would seem, an area escapes over a
period which permits it to become intensely infective, and an outbreak
of yellow fever results.

The localised character of these outbreaks in a town is due to
the well-marked domestic habits of the mosquito concerned. If we
look at the spot maps accompanying the Reports of the 1910-11
epidemic, it is easy to see the human agency which carries the disease
from one part of a town to another, over distances which leave
intermediate areas unattacked.

It is, therefore, the infected rather than the infective element

437

which is responsible for the spread of the disease. If we can control
the former and keep it from coming into close living contact with the
unsegregated European, there is a reasonable prospect of preventing
re-occurrences of these outbreaks. Efforts at controlling the other
element have hitherto met with very little appreciable success. That
temporary success 1s obtainable has been amply demonstrated during
the present epidemic. “Towns where the Stegomyia index is normally
80 per cent. have, after a week’s intensive work, had this figure
reduced to below 5 per cent. The means employed, other than
fumigation of the area in which cases occurred, were the usual
mosquito brigades under the supervision of European volunteers.

The effectiveness of this measure, when considered in the light
of what has already been said, suggests the advisability of instituting
a ‘cleaning-up week ’ at least once a year, in every town where cases
of yellow fever have been known to occur within the last twenty
years. It should also be a matter of routine that when a case of
yellow fever is reported in a Colony, every town with which the
infected area is connected by road. rail, or sea, should immediately
start energetic anti-stegomyia measures. This will prevent outbreaks
elsewhere, for the reason that the number of mosquitoes remaining
will not be able to maintain the disease. At any rate, the possibility
of a secondary focus being established will be a very remote one.
In the intervals between epidemics, Government Medical Officers and
other Medical practitioners should be asked to observe carefully cases
of fever which occur in West Africans who are strangers in the place,
with a view to early diagnosis, thus ensuring prevention of the
development and spread of the disease.

REFERENCES

Gurreras, Juan (1921). Observations on Yellow Fever in a Recent Visit to West Africa.
Sanidad y Beneficencia, Vol. XXV, p. 21.

Horrmann, W. H. (1921). Das Gelbfieber in Westafrika. Arch. f. Schiffs-u. Trop.-Hyg.,
Vol. XXV, p. 291.

Peterson, E. (1922). Yellow Fever in St. Thomas, with special reference to its spontaneous
elimination. U.S. Naval Medical Bulletin, Vol. XVII, p. 555.

Report on Certain Outbreaks of Yellow Fever in 1gto and 1911. Waterlow & Sons, London
(1913).

Yellow Fever Commission (West Africa) Fourth and Final Report. J. and A. Churchill,
London. (1916), p. 255.

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439

MISCELLANEA"

KURLOFF BODIES IN FISH

In a specimen of Diodon hystrex examined at Freetown, Sierra
Leone, Kurloff bodies were found in about 70 per cent. of the
lymphocytes. As many as five Kurloff bodies were present in
some cells.

S. ADLER and E. J. CLARKE.

TRICGHONEMA TETRACANTHUM (Meu tts, 1831,
oF Looss, 1900)

This worm was found by us in June, 1923, in a donkey born
and bred in the north of Ireland. This record is of interest, as
the parasite has not been found since it was described by Looss in
1900. The fact that it had not been observed in Europe is used by
Railliet (1923) as an argument that Looss’ parasite is not identical
with Sétrongylus tetracanthus, Mehlis, 1831.

J. W. S. MACFIE and WARRINGTON YORKE.

PIGS AND ANKYLOSTOMIASIS IN THE
GOLD COAST

During January and February, 1922, forty-eight pigs were
examined at the Accra slaughter-house for hookworms, but
neither Axcylostoma duodenale nor Necator americanus were found,
although 3,270 other small nematodes were collected, the majority
of them being Cesophagostomum dentatum (Rud., 1803), and a few

*It is proposed to publish under this heading short records relating to Tropical Medicine
and Parasitology.

440

Arduenna strongylina (Rud., 1819) and Characostomun longe-
mucronatum (Molin, 1861). Subsequently specimens obtained from
pigs at Cape Coast, at Kumasi, and at Sekondi (Dr. J. F. Corson)
were examined, and in these also neither A. duodenale nor
N. americanus was found. These results do not, therefore, support
the view that pigs are an important factor in the dissemination of
hookworm infections in the Gold Coast. —

J. W. S. MACFIE.

ONCHOCERCA ARMILLATA IN CATTLE
IN THE GOLD COAST

Commes and Devanelle (1917) record that in Upper Senegal and
Niger, Onchocerca armillata, Railliet and Henry, 1909, is a common
parasite of cattle, and that they found it in one hundred and
fifty-one animals out of one hundred and ninety-eight, that is in
760°3 per cent. It is also a common parasite of cattle in the Gold
Coast, particularly of the hump-backed breed, as is shown by the
fact that of forty animals examined at Accra during September and
October, 1922, namely, sixteen hump-backed cattle and twenty-four
of the straight-backed breed, fourteen, equal to 87°5 per cent., of the
former, and seven, equal to 292 per cent., of the latter, were
infected.

The situations in which the worms were found and the lesions
(atheroma, calcification, cyst and nodule formation, etc.) associated
with them were similar in the Gold Coast cases to those described
by Commes and Devanelle, and need not be referred to in detail.
Some of the nodules contained, in addition to a mass of fibrous
material and portions of parent worm, a number of free larvae. The
larvae resembled in general form those of O. volvulus, length of the
few measured 280u to 345m, breadth about 5m, anterior end
rounded, nerve ring well marked and situated at about 25 per cent.
of the length from the anterior extremity, and tail sharply pointed.

In three infected animals, the blood (10 c.c. or more) was
examined for larvae, but without success. In this connection it
may be recalled that in blood films from one hundred and sixty-six
cattle examined at Accra in 1914, filarial embryos were found in five,

441

that all these were sheathed and were perhaps embryos of Sezarza
labiato-papillosa, a species which has been found in cattle at Accra
(Macfie, 1915), but that no larvae resembling those of O. armillata
were encountered. The skin of these three animals was also
examined, because it was thought that, as in the case of O. volvulus,
larvae might be present in it. No larvae were found, but it
must be admitted that considerable difficulty was experienced
owing to the thickness, density, and hairiness of the skin, and that
consequently the examination was not a very satisfactory one.

Tea Wee. MACFIE.

REFERENCES

Comes, C., and Devanetie, P. (1917). L’Onchocercose aortique bovine dans le Haut-
Sénégal-Niger. Bull. Soc. Path. Exot., Vol. X, p. 459; with Note by RarLuier, p. 461.

Macriz, J. W.S. (1915). Report of the Accra Laboratory for the Year 1914, p. 34. J. and A.
Churchill, London.

Raruet, A., and Henry, A. (1912). Nematodes vasculicoles des Bovins annamites. Bull.
Soc. Path. Exot., Vol. V, p. 115.

443

A STUDY OF THE TUMBU-ELY,
CORDYLOBIA ANTHROPOPHAGA
Grunzerc, IN SIERRA LEONE

BY
By BACK LOGK

AND

M. G. THOMPSON

(From the Sir Alfred Lewis Jones Research Laboratory,
Freetown, Sierra Leone.)

(Received for publication 15 August, 1923)
PLATES XV—XVIII

TABLE OF CONTENTS

PAGE

I. INTRODUCTION ... ae re He 405 = os "oe dee soe ee Aaa

II. NOMENCLATURE Ae a aes a TT Yr a sae coe 1446
TI. GeroGrapuicat DisTRIBUTION eae se ae Ae ae Sais area 4 7

IV. MorpHoLtocy AND Bionomics—
(ryiAXehalt it . 2 ae. ¥3 Sec ae oe +7 es 2 i. 9449
(2) Egg is ome 2 SA eee eee ee eee ge er ers

(3) Larva—
(ae weehistar 4202 BIBL? STON FE DA, 957 14 S79
(6) 2nd instar cas) Meceate. «9h meee oe ae > 2 Fy es tinh eaeh 9 409
(c) 3rd instar aes ose eae re ee we ms ee Aa

(4) Puparium eee ee Sern eet | Sse ee ee AT
V. DeveLOPMENT IN ANIMALS—

(1) Duration in experiments Se See er on cted abies op een

(2) Pathogenicity to animals =e es “yt rahe nie bee ATO

(3) The animal hosts in nature... oe 3% = aii ak if: GEaSs

(4) The main natural reservoir... ae ve ane ee Pee gh y-

(5) Temperature of animal host and rection ow Ie ook ask 2 dod

(6) Puparia in nature ot an 8 & 7m Bie 32 HICSS

VI. Ace INcIDENCE ask aoe. BAF aoe er as bed oe See vA. O0
VII. Immunity pias +e ? “ap 5ee Bas ae i Sey ack AOD
VIII. Srasonat INCIDENCE ... eee wee ane are tr ae <s peu iitag2
IX. Mops or InrecTtion or Man uv. waa ae ae eee seh 228 ).404.
X. SYMPTOMATOLOGY ... a AY: was ee nae rom aoe cos AOS
XI. TREATMENT... “evs 2 ae eon ce aoe Ay ag a Ag
XII. Propuyraxis ... ae isi it. 2 rae ie sop) 498
XIII. ComparisoN WITH OTHER CUTANEOUS Mare Decgeenee FL i&s Res weet 490
XIV. Summary pee ‘i oe Ser ae aoe ae tee eae eae ee SOL

XV. REFERENCES... ae ah: wide oa. Lae as Pp ate Bord set

444
I. INTRODUCTION

Although it is more than sixty years since human Myiasis due
to the larva of C. anthropophaga was first investigated by two
French naval surgeons in Senegal, it 1s a remarkable fact that even
to-day complete unanimity as to the mode of infection, the seasonal
incidence, the natural reservoirs of the infection, and the appropriate
prophylaxis, 1s by no means attained. It is interesting to find, in
respect to this fly and also the South American fly, Dermatobia
cyaniventris, that a disregard of native accounts has led research
into side tracks which have been followed for long periods, before
it was discovered that they were leading in the wrong direction.
Nothing can better illustrate the manner in which this has occurred
than the following extract from the excellent account of Cordylobia
Myiasis given by Coquerel and Mondiere (1862), who published a
paper—the first so far as is known at present—on this subject.
Writing of this form of Myiasis as observed by them at Portudal
in Senegal, they say ‘Cette singuliére affection est connue des
indigénes, qui savent tres bien extraire les larves qui les tourmentent
et viennent souvent se loger dans les tissus du scrotum de ces
malheureux. Ils prétendent que ces vers sont produits par une
petite mouche trés commune a Portudal. Cette mouche pondrait
ses ceufs dans le sable humide, le ver y sejournerait jusqu’au moment
ou profitant du repos d’un homme étendu sur le sol, il s’introduirait
dans la peau de sa victime.’

Unfortunately the authors were so little impressed by the -
probability of this suggestion that they added ‘ Il n’est pas besoin
d’insister sur les détails de ce récit pour en signaler les erreurs.
Il est évident que les larves du Diptére du Sénégal ont été déposés
dans la peau, ou que les ceufs ont été fixés a quelque poil de cette
membrane des leur origine et que les vers ne peuvent vivre ailleurs.’
The theory held by natives in Senegal in those days with regard to
the mode of infection by the larvae of Cordylobia, is the same which
is held to-day by natives of Sierra Leone, and it is the theory which
has been proved to be, in all essentials, correct, by the accumulated
research of European observers up to the present day.

In a similar way we find that the work which has been done on
the bionomics of D. cyaniventris in South America has revealed the

445

interesting fact that the eggs of this species may be transported on
the body of a mosquito and deposited on the animal which is to
serve as the host of the larva. It has taken many years to produce
the scientific evidence that such a remarkable means of infection can
occur, yet the native peasants have known it for so long that their
name for the larva is Gusano de Zancudo, 2.e., the worm of the
mosquito. The suggestion of such a means of infection was treated
with frank incredulity by such observers as Da Silva Aranjo, who
refers to the peasants’ belief in the transference of Dermatobia
infection by mosquitoes as ‘a popular error, very widely spread
throughout Brazil.’ This incredulity of those who first investigated
the bionomics of Dermatobia had the same effect as that mentioned
above, in delaying the discovery of the fact that the mosquito,
Janthinosoma lutzt, actually does transport the eggs of Dermatobia.

The suggestion has been made by Zepeda (1913) that the larvae
of Cordylobia may be carried in the same way by mosquitoes; we
have found no evidence of this.

The accurate knowledge displayed by the intelligent African
native, the uneducated native who uses his powers of observation to
the fullest extent, is often impressive. The Protectorate natives of
Sierra Leone quite commonly make for example the nicest distinction
between the bionomics of the testaceous flies, C. anthropophaga and
A. luteola. They recognise that the flies are very similar to each
other in appearance, and that each produces eggs out of which larvae
proceed ; they are perfectly aware, however, that of one, the larva
lives in the tissues of the host and produces boils, while of the other,
the larva merely feeds on the cutaneous blood of the host and lives
in the ground, only emerging at night. They can also procure
samples of either larva in a short time if required to do so, the
former expressed from the larval tumours in the skin of affected
animals, the latter obtained from the earth on the floor of their huts
by the simple procedure of sleeping on a mat on the floor and
searching under the mat in the early hours of the morning. It 1s,
therefore, no more than justice to recognise how in Myiasis, as in
very many other diseases, the knowledge of the natives, acquired by
the slow and painful process of racial and personal experience, has
assisted the investigator to the correct solution of the problems of

disease.

446
Il. NOMENCLATURE

From the earhest days of our knowledge of Myiasis due to
Cordylobia, that is to say, from the time of the publication of the
communication of Coquerel and Mondiére in 1862 until the most
recent times, when in 1914 Roubaud published his well-known
treatise on Myiasis in French West Africa, very various opinions
have been held as to the probable relationship which existed between
larvae from diverse hosts in different parts of Africa. Coquerel and
Mondiére, who were not fortunate enough to rear the adult fly,
considered the larva to be that of an Oestrid.

Bérenger-Féraud (1872) was successful in rearing flies which
were identified by Emile Blanchard as belonging to the genus _
Ochromyia, Macq.; Blanchard called them by the new specific
name, Ochromyia anthropophaga. Of this name Austen (1907)
says, ‘ Since, however, no description of the fly whatever was given,
Ochromyia anthropophaga, Emile Blanchard, is a mere omen
nudum, and consequently invalid.’ ‘The association of the name
Bengalia depressa in 1891 with larvae from a human case of Myiasis
has proved an additional complication, and still remains so to-day
in some publications, in spite of Griinberg’s work. Austen, in the
paper mentioned above, gives ‘a statement of how the larvae of
human Myiasis due to Cordylobia came to be associated erroneously
with the adult fly Bengalia depressa, Walk. He points out that
the type of B. depressa is in the British Museum, and that although
it is an allied, nevertheless it is a very different, insect from
Cordylobia; he says, moreover, that the life history of b. depressa
is as yet unknown, and that there is not a particle of evidence to
prove that its larva is a subcutaneous parasite.

It was Griinberg (1903) who, after a careful examination of all
the material available, both larval and adult, came to the conclusion
that the fly did not belong to the genus Ochromyia, nor yet to
Bengalia nor Auchmeromyia; but was a fly which required for its
classification a new genus; he accordingly erected the genus
Cordylobia, with the species C. anthropophaga (Blanch.). He gave
a long and detailed description of genus and species. D6nitz (1905),
in an article entitled ‘Uber eine neue afrikanische Fliege mit
parasitisch in der Haut von Ratten lebenden Larven,’ gives a
description of what he considers to be a distinct species of

447

Cordylobia, and names it C. murium. At the same time, Donitz
reviewed the position with special attention to the ideas expressed
by Griinberg and Gedoelst (1905), and shows how the latter came to
speak of a Cordylobia anthropophaga, Griinberg. Déonitz himself
proposed the name Cordylobia grinbergi for the East African form.

Roubaud (1914) gives reasons for deciding that Cordylobia
murium should not be retained as a separate species, namely, that
the differences claimed by Donitz to exist are not sufficiently striking
or constant to warrant the species; at the same time the name
Cordylobia grinbergi is dismissed by him as invalid.

Two other species also are dealt with by Roubaud; of Cordylobia
praegrandis, Austen, he says that the subsequent discovery of the
male has shown that it does not belong to the genus Cordylobia but
should be placed in the genus Chaeromyia, Roubaud, while
Cordylobia rodhaini should likewise be placed in another genus.
To sum up, Roubaud says there appears to be only one species of
Calliphorines belonging to the genus Cordylobia as defined by
Griinberg; that is the fly of Cayor which was first bred by
Béranger-Féraud in Senegal, Cord ylobia anthropophaga, Blanchard.
Austen, however, in his summary of the situation, referred to above,
ends with the remark, ‘The correct designation of this highly
important and much misunderstood African Muscid is, therefore,
Cordylobia anthropophaga, Grinberg. This authoritative statement
we have, therefore, accepted.

III. GEOGRAPHICAL DISTRIBUTION

Griinberg (1903) gave as the distribution of C. anthropophaga
a list of places, which included Senegal, South-West Africa, Gaboon,
Dar-es-salam, Zambesi, Lake Nyasa, Tanga, Delagoa Bay,
Bagamoyo and Durban. The distribution later given by Roubaud
is from Senega! and Lake Chad to the Cape. In the map included
in his work, practically the whole region of Africa, south of about
16° North latitude, is shown as infected, with the exception of the
North-East area. He points out, however, that the fly is irregularly
distributed and that many large areas so far appear to be free of it,
or that it has not been recorded from them.

448
OCCURRENCE OF C. ANTHROPOPHAGA IN SIERRA LEONE

Many observations of the clinical effects produced by the larva
in man and animals in Sierra Leone have been made in recent years,
and this Colony has for many years been considered to be a favourite
haunt of Cordylobia. Smith (1908) remarks that ‘Tumbu’ is a
Negro-Creole word, and gives a record of his findings of the larvae
in Sierra Leone. He expresses doubt as to the correctness of the
accepted mode of infection by the laying of the eggs or larvae in the
skin; this doubt arose from his observations of the situation of the
lesions in animals. Blenkinsop (1908) noted that in Europeans the
upper part of the thigh and the buttock are the favourite site for
the larvae to gain an entrance, and it is a generally received opinion
that the parasites are often acquired at the latrine. The West
Indian troops were often affected in the axilla, and natives, in any
region. It was not known in 1908 whether Cordylobia was
Oviparous or viviparous, but Austen said that in either case, since
the female is undoubtedly unable to pierce the skin with her
ovipositor, the larva in its earliest stage must bore its own way
through the integument by aid of its mouth hooks.

Smith made observations, as we shall see later, on the age
incidence of the disease in man and animals, and also succeeded in
breeding out flies from larvae obtained from rats and dogs. He
mentions a wild rat which had six ‘tumbus’ in the bare underpart
of its legs and feet, which were immensely swollen.

The prevalence of Cordylobia Myiasis in Sierra Leone is so
considerable—the parasite itself is the cause of much discomfort to
man, and causes suffering and even death in animals—that we took
the opportunity of studying as carefully as possible the bionomics
of the fly in its various stages, and of making experiments with a
view to discovering the best method of attack upon it. We have
also made observations upon the morphology of the first instar
which may throw some light upon the mode of skin penetration of
the larvae of other forms of Myiasis, notably those due to
Dermatobia ; owing to the fact that we found that the various stages
did not always correspond to previous descriptions, we have
included a short description of each of the stages of the fly. It is
necessary for us to refer frequently to the work of Roubaud, as his

449

is the most recent and at the same time the most comprehensive
work done on the subject. We have been enabled, largely owing to
the greater amount of material at our disposal, not only to confirm
his observations in many particulars, but also to add to them. If
we are compelled to differ from him in a number of points, it is due
entirely to the fortunate circumstance that a larger amount of
material enabled us to carry our experiments further than he was
able to do.

IV. MORPHOLOGY AND BIONOMICS
(1) ADULT (See Plate XV)

The following description of the adult stage is taken from
Austen (1908): ‘A thick-set, compactly-built fly of an average
length of about 9} mm.; specimens as small as 64 mm. or as large
as 10 mm. in length are occasionally met with. Head, body and
legs, straw yellow; dorsum of thorax and of abdomen with blackish
markings; wings with a slight brownish tinge. The eyes meet
together for a short distance in the median line above in the case of
the male, but are separated by a broad front in the female.
On the dorsum of the thorax the dark markings, which are
a pair of longitudinal stripes not reaching the hind margin, are
covered with a greyish bloom, and, consequently, not very
conspicuous ; this bloom 1s also present on the abdomen, but here the
markings are much more distinct, especially in the female, 1n which
the third segment, as also the fourth segment with the exception of
the hind margin, is entirely black or blackish. In the female, the
second segment is marked with a blackish quadrate median blotch,
and has a similarly coloured hind border, broadening towards the
sides, while the first segment has a narrow dark hind margin. In
the male, these markings are not so extensive; the dark hind margin
to the second segment is interrupted on each side of the median
blotch, which is triangular in shape, and there is a yellow area of
considerable size on the proximal half of the third segment, on either
side of a blackish median quadrate blotch; the fourth segment is
similarly but less conspicuously marked,’

450
HABITS OF WILD FLIES.

The adult fly material which most observers have had at their
disposal has, as a rule, been obtained by the process of breeding
flies in the laboratory from larvae taken from the furuncular tumours
of animals. Reference in the literature to the capture of adults is
extraordinarily meagre. Rodhain and Bequaert (1913) observed
wild adult females flying round the cages in which animals were
kept, and followed the egg-laying process. Eggs were deposited in
the straw and manure in the cages; experimental animals, monkeys
and guinea-pigs, placed in the cages where the wild flies had laid,
became infected, as the result of the larvae which emerged from the
eggs penetrating the skin. In 1911 Roubaud, at Bamako, captured
alive one fertilized female which laid eggs in captivity, and which
supplied the egg and larval material for his experiments.

We have been exceptionally fortunate in this respect, because we
have at Freetown, Sierra Leone, notorious in the history of Tumbu
disease, been able to capture many adults indoors. Not only were
numerous adult females and males. captured, but several of the
females were either fertilized before capture, or were fertilized after
capture without difficulty. A point of interest is that these captures
were effected and the experiments resulting from them were carried
out in the dry season, during the months November, 1922, to April,
1923; in its proper place, further reference will be made to the
bearing of this fact on the seasonal incidence of Myiasis due to
Cordylobia.

On occasions the wild flies were seen on the wing; for example
on the 27th March at sunset, on a cool evening several flies were
seen in the open darting about after each other and buzzing loudly ;

they dashed into objects blindly, and one, a female, which had
injured itself in this manner was captured.

Natives were able at times to capture adults in their houses, but
the construction of their houses taken in conjunction with the resting
habits of the fly as observed by us, explains the lack of success which
often attended the efforts of the native to capture flies. The flies
captured by us were found resting on the dark green painted ceiling
of the bungalow verandah; on bright sunny days as many as three
or four would be found there; on cloudy days they were rarely

451

found. “They would remain there motionless for long periods, and
only when disturbed would they fly about with great rapidity; they
emitted during flight a loud buzzing noise, similar to that produced
by the blow-fly; the noise ceased when they alighted again.
Against the dark surface they presented a very inconspicuous
appearance, and would commonly be overlooked. It was easy to
understand that if this method of resting were followed on the
smoky roofs of houses of native construction, the fly would be even
less conspicuous. ‘The flies were easily caught with a collecting net,
and they gave the impression of being unable to see well in the day,
as they allowed the net to approach close to them without taking
flight. Wald flies were seen twice at night attracted by the light
of a lamp on the verandah; they flew round noisily, knocking
themselves against the lamp, and several had their wings scorched
and fell inside the chimney. Whether these flies had come from
out of doors to the ight or had come to it from some resting place
indoors is uncertain. It 1s probable the latter is the case, as after
the systematic capture of all flies resting indoors in the daytime
had been undertaken, no further captures were made at the lamp
at night.

Wild or laboratory bred flies when placed in glass containers
such as cylinders or inverted bell jars, of which the upper end was
closed with cloth, rested chiefly on the cloth in the same upside-
down attitude as did the wild flies on the ceiling. During the day
they were rarely on the wing, but in the early morning from seven
to nine, and in the late afternoon from four to six, they became
very active, flying about, striking the glass sides of the vessel and
buzzing audibly. At night they rested much as in the day, but the
appearance of light near them at once aroused them to great
activity.

FOOD oF ADULTS.

Roubaud observed a wild female fly feeding on sugar, on pulped
fruit, and on ground soiled by urine. We found that both males
and females, whether wild or bred, fed readily on banana and
pineapple, the females feeding longer and oftener; both sexes also
sucked up the juice from pieces of decomposing rat liver, and less
readily fresh blood of a rat from a drop exposed on a slide.

452

RESISTANCE OF ADULTS IN VARIOUS CONDITIONS.

Direct sunlight. Three wild flies were exposed in large test-
tubes to the direct rays of the sun during the hours I! a.m. to
2 p.m. They survived only from fifteen to thirty minutes. The
results of this experiment serve to explain the fact that on bright
days during the hot hours the flies come indoors to rest.

Dry heat. Wild flies were exposed to varying degrees of
heat ; they were placed singly in wide test-tubes which were plugged
with wool and provided with a thermometer, the tubes were placed
in a water bath which was rapidly brought up to the desired
temperature. A male kept for thirty minutes at 44-45°C. was
still active at the end of the period. On raising the temperature
rapidly great restlessness was observed at 50° C., and at 52°C. the
fly dropped suddenly dead to the bottom of the tube. The
experiment was repeated with two other wild flies, one male and
one female. The temperature was raised from 40°C. to 47°C. in
two minutes; after one minute at 47°C. both fell dead suddenly.
It appears from these experiments that a temperature of about
50° C. 15 fatal for they.

Cold. Your laboratory bred flies, one male and three females,
were enclosed in a large tube containing a slice of banana and
placed in an ice chest on 16th March, 1923. After an hour in the
ice chest at a temperature of from 10° C. to 6° C. all were motionless.
Two were remnoved to room temperature and rapidly recovered ; they
were then returned to the ice chest. The flies at each subsequent
examination were motionless, sitting sometimes on the glass and
sometimes on the banana; they were not observed to feed but
changed their position slightly, and recovered their feet when
shaken down. On 20th March, 1923, the one male died; the three
females died on 23rd, 24th and 28th March, 1923, respectively.
The powers of resistance to cold and damp are, therefore, very
considerable. |

OVIPOSITION. As we have seen, Rodhain and, Bequaert observed
Oviposition on straw and manure in animal cages. Roubaud noted
that his fly laid eggs on the glass walls of a vessel and on fruit.
We found that of various sites on which gravid females in captivity
were given the opportunity of depositing their eggs, the one most
commonly selected was dry sand which had previously been

453

contaminated by the excreta of animals, in this case guinea-pigs.
This fact was observed on an occasion when three females were
placed in a bell-jar containing a guinea-pig in order to determine
whether they would deposit eggs on the animal’s skin. These flies
had up to this moment been lodged in a container where they had
access to cardboard, cotton wool, banana, and glass on which to
oviposit; none of these sites was apparently suitable for them, as
they did not utilize them. Immediately on their being admitted
to the bell-jar containing the guinea-pig, two, and after a short
delay, the third also, set about depositing their eggs in the
contaminated sand with great eagerness and rapidity; eggs were
not laid by them on banana leaves, carrot or orange, which were
also present. It is perhaps not without bearing on this point that
three other females in which ova apparently mature were present,
died in the first container without laying their eggs. On three
occasions, when sand was available and utilized, eggs were also
laid on sites other than sand, but only in small numbers, viz., six
on a piece of black cloth, seven on the white cloth cover of the
bell-jar, and eleven on cotton wool. On only one occasion was a
considerable number of eggs laid on any other material than sand,
when contaminated sand was available; this was a case in which wet
sand had been provided for the fly; she landed on it and protruded
her ovipositor, but apparently found it too wet for her, as she
immediately flew off; she laid one hundred eggs in a plug of pink
cotton wool which was used as a stopper to the central aperture of
the white cloth cover. It appears probable that the result of this
experiment has some significance in regard to the wet seasonal
incidence of this form of Myiasis in man and domestic animals.
Apart from these occasions eggs were always laid in the sand
provided for the guinea-pigs. In numerous experiments conducted
during the laying of hundreds of eggs, flies could not be induced to
leave the sand on which they were laying. The guinea-pigs did
not attract them, nor did they oviposit on clean cloth nor on cloth
impregnated with human perspiration, the pieces of cloth being
placed in their path as they were laying their eggs. Flies on the
other hand would not oviposit on sand contaminated with excreta,

if the sand was too moist.
Method of oviposition. Generally for some hours, even a day,

454

before egg-laying commenced, the female could be seen pushing out
and withdrawing the ovipositor, and from time to time small drops
of clear fluid appeared at its tip. The procedure when ovipositing
in contaminated sand was uniform for all the flies observed. The
fly, having alighted on the surface of the sand, and having found
a suitable area, digs with the tip of the abdomen a small cavity in the
sand, backing slightly and curving the abdomen downwards to
enable it to do so; the ovipositor is then extruded and pushed into
the sand at the bottom of the small cavity. At this time, when the
ovipositor sinks into the sand, the two hind legs bring up on either
side a few grains of sand against the ovipositor, which is then
withdrawn. The hind legs next move ‘rapidly in a _ horizontal
direction to scrape a little sand over the egg deposited in the small
cavity, and to smooth the surface. The fly then advances hurriedly
a few steps and commences again to dig in the sand, and repeats
the whole process; she does not move in a straight line for long,
but turns in her tracks frequently, with the result that a small area
may be very thickly sown with eggs. The movements of the fly in
the later stages of egg-laying often disturb eggs previously laid by
it and uncovers them, bringing them to the surface. On cotton wool
the eggs were laid on strands about one quarter-inch from the
surface.

Batches and number of eggs laid. From the fact that his fly,
which laid over one hundred and fifty eggs, died after ovipositing,
Roubaud concluded that Cordylobia cannot survive parturition; also
he concluded that probably, as the number of eggs laid was much
higher than what he found in the case of Auchmeromyia, only one
batch of eggs is laid by Cordylobia. Our observations show that at
least two batches of eggs may be laid, and that the female does not
die immediately after parturition. For example, a wild fly, No. 22,
was observed to lay two batches of eggs in captivity. It was
captured on 20th January, 1923, oviposited on Ist February, 1923,
and again on 11th February, 1923; it died on 16th February, 1923.
A laboratory bred fly, No. 38, emerged from the pupa on
23rd February, 1923, and was fertilized while still unfed on the
same day by a wild male; she laid the first batch of eggs on
5th March, 1923, and a second on 8th March, 1923; she died on
10th March, 1923. Another laboratory bred fly which emerged on

455

roth March, 1923, and was fertilized on 11th March, 1923, laid a
first batch of eggs on 17th March, 1923, and a second on
20th March, 1923; she died on 24th March, 1923. The number of
eggs laid in the first batch varied from two hundred and eighty-
seven to three hundred, in the second batch from ninety-four to
one hundred and eighty-four. It appears probable that the batch
of eggs laid by Roubaud’s fly was the second batch.

Dissection of gravid females. Several females which died
without laying eggs were examined. One laboratory bred female
which emerged on 20th February, 1923 and copulated on the same
date died on 3rd March, 1923, without laying; she contained three
hundred and four eggs; another laboratory bred female which
emerged on 10th March, 1923, and was not seen to copulate, died
on 24th March, 1923, containing five hundred and three eggs in
different stages of development; a wild fly, with which a wild male
would not copulate, died on 23rd February, 1923; she contained
four hundred and four eggs.

Rate of Oviposition. On several occasions when females were
engaged in laying in the sand, the total time taken in laying a batch
of eggs was noted, and also the rate per minute. The total time
taken by one fly in laying two hundred and eighty-seven eggs was
thirty-three minutes; by another for one hundred and eighty-four
eggs was twenty-six minutes. During the time there were several
pauses of varying length, and this reduced the average number of
eggs laid per minute. On the whole, however, the rate was very
constant for all the flies observed. Thus, taking a total of forty-six
individual minutes timed among several flies at the time they
were ovipositing, the smallest number of eggs laid in a minute was
five, the highest eleven, the average per minute being eight. This
relatively slow process is against the idea of egg-laying on animals.

LENGTH OF LIFE OF FLY IN CAPTIVITY.

The longest period during which a wild fly lived in captivity
was eighteen days; this was a female, which during that period laid
two batches of eggs, surviving the last oviposition for five days.
Several laboratory bred females lived fourteen days, but only one

lived for fifteen days.

456

(2) EGG

This is white in colour and measures on an average o'8 mm.
in length; it is banana shaped, being almost straight on one
side and curved on the other; it tapers somewhat towards one end.
On the surface there are longitudinal grooves, and there is also a
fine hexagonal reticulation. In eggs from which the larva has
emerged it 1s seen that there is near the smaller pole a longitudinal
slit extending about one-third along the flattened surface; through
this slit the larva has emerged. |

SITE. The eggs were found just under the surface of the sand
in which they were laid; in cotton wool also, not on the surface but
about a quarter of an inch deep. The eggs adhered in most cases
to particles of sand, or in cotton wool to the strands, and could not
be shaken off. i

HATCHING. For some hours before hatching the egg shows a
darker patch towards the more pointed end; as the time of hatching
approaches it is seen that this dark patch is in active movement,
and it is recognised as the chitinous buccal armature of the larva
tearing at the inner surface of the eggshell. By means of this
armature the larva cuts a linear opening on the flat surface of the
egg near the small pole; as soon as it is possible to do so, it pushes
its cephalic end through the aperture, which it proceeds to enlarge
by vigorous movements of the anterior body segments. In cases
watched throughout the process, it usually took from four to six
minutes from the moment when the aperture was first observed till
the time when the larva had cleared itself of the eggshell.

RESISTANCE OF THE E.GG TO VARIOUS AGENTS.

Room temperature. On glass the larva emerged in three days
as a rule. Roubaud found a shorter period on sand, and noted
that eggs on wet sand hatched somewhat later than eggs on dry
sand.

Incubator at 37°C. If eggs were placed in watch-glasses either
dry or immersed in a small quantity -of water they hatched in
twenty-four to forty-eight hours, the water drying up.

Sunlight. Exposure to the rays of the sun for one hour, whether
on glass or on dry or wet sand, did not prevent them hatching
within four days. Two larvae in this experiment were watched

457

leaving the egg; the process was short, less than a minute elapsed
before the larvae were delivered from the egg, but they dragged the
eggshell about for another half minute, before getting rid of it.
Eggs exposed to the sun for four days did not hatch, even when
subsequently removed to the shade at room temperature.

Dry heat. Numerous experiments were carried out with eggs -
in plugged tubes placed in a water bath. Exposure to temperatures
of 60° C., 55° C., 50° C., and 45°C., for two minutes killed all eggs
used.

Wet heat. Similar results were obtained by heating eggs
submerged in water in tubes placed in a water bath at these
temperatures for two minutes.

flot woning. Eggs were rendered incapable of hatching by
passing over them lightly a flat iron at a temperature suitable for
pressing clothes. ‘The eggs were not protected by being in cotton
wool, nor even by three folds of cotton cloth; they were flattened
and desiccated in the process, a point of some practical importance
in view of the frequently accepted theory that clothes are infected
when at the laundry.

Cold. Eggs placed in an ice chest did not hatch in seven days,
nor after removal to room temperature. Four eggs were placed in
the ice chest for forty-eight hours; on removal to room temperature
two emerged in three days.

Eggs dissected out of dead females did not develop in any
medium, either at room temperature or at 37° C. in the incubator.

(3) LARVA

First INSTAR. Many descriptions of larvae from cases of
Myiasis have been made from larvae which were in the later stages
of development. It is, however, very important that not only the
later instars should be examined, but that the first instar should
receive attention, At this stage, in those larvae which can produce
true cutaneous Myiasis, very interesting adaptive structures are
found, some of which appear to render the larvae capable of
penetrating unbroken skin, while others determine the ability of the
larvae either to remain in situ in the skin or to penetrate further
into the tissues. The structures which attract attention chiefly are
the cephalo-pharyngeal skeleton and the cuticular spines. The first

458

stage larva of Cordylobia presents points of interest in respect to
both these structures, as will be seen in the description given below.
Phenol was used as a clearing agent.

The newly hatched larva is white in colour and is visible to the
unaided eye. It measures from 0°75 mm. to I mm. in length; it is
somewhat fusiform, tapering from the mid region towards the
anterior, and to a less degree towards the posterior extremity; it is
composed of thirteen segments. The first or cephalic segment is
the smallest ; the mouth aperture 1s situated near the ventral surface
of this segment; the ventral surface of the segment adjacent to the
mouth is yellowish, chitinized, and densely clothed with yellow
spines directed backwards. On the dorsal region of the segment
there are anteriorly two rounded projections, one on either side.
On the posterior portion of each projection there is a minute
antenna-like structure consisting of two segments: on the anterior
portion is a small chitinized pit. Near the caudal margin of the
segment are several rows of backwardly directed yellow spines.
Segments number two to eight are covered with backwardly directed
spines, almost colourless except towards the cephalic margin of each
segment where the spines are distinct and more heavily chitinized,
yellow or even brown in colour. Segment nine is almost devoid of
spines, being provided at the cephalic margin with a single row of
backwardly directed spines and at the caudal margin with a single
row of spines, in this case forwardly directed. Segments ten and
eleven have no spines at the cephalic margin, but on the caudal
margin have several rows of spines directed forwards, the rows being
more numerous on the dorsal aspect.

Segment number twelve is longer on the dorsal than on
the ventral aspect; it is densely clothed all over with large,
strongly chitinized and forwardly directed spines. ‘These large
spines directed forwards and strongly chitinized appear to act in
keeping the larva in position with its posterior end at the surface
of the skin. It is interesting to note that, judging from the
drawings in Surcouf’s (1913) article of the first instar larva of
Dermatobia cyaniventris, a similar arrangement exists there. This
segment in Cordylobia is furnished with several soft digital
processes ; of these two are visible on the dorsal surface, one on either
side of the middle line, two are situated laterally on the segment,

459

one on each side, while two are situated on the ventral surface, one
on each side of the anal orifice.

Segment number thirteen is small and has only a few sparsely
distributed spines; on this segment there are four pairs of soft digital

Stesct3¢)

Fic. 1. (1) Egg hatching. (2) First instar larva. (3) Head of larva of first instar, ventral
view. (4) Head of first instar larva, lateral view.
A—antenna, later papilla; B—chitinous pits ; C—prestomal sclerite ; D—median
buccal spine ; H—oral rods; #—cephalo-pharyngeal sclerite ; G—ventral spiney area
adjacent to mouth.

460

processes in addition to the tracheal tubes; the tracheal tubes open
dorsally on the segment near its anterior margin on flattened
eminences. The digital processes have in many cases at their tips a
chitinous pit; the processes are of assistance to the larva in locomo-
tion, and it is by means of them that the larva can attach its posterior
extremity to particles of soil or other objects on which it rests and
holds itself erect in the air, while waving its anterior end about in
search of a host. The arrangement and position of these processes,
fourteen 1n all, on the last two segments can be seen in figure 4.

The buccal armature or cephalo-pharyngeal skeleton 1s adapted
for penetrating unbroken skin. It consists of a median buccal spine
heavily chitinized, which articulates posteriorly with the cephalo-
pharyngeal sclerites. On either side of the spine are placed the
feebly chitinized oral rods (baquettes orales of Keilin). The
cephalo-pharyngeal sclerites are long and slender, and have poorly
developed dorsal and ventral cornua. The median buccal spine has
received various names, for example, median hook, median tooth,
and labral sclerite; according to Keilin, ‘ the median hook occurs
in the primary larva of nearly all Cyclorraphous Diptera.’ In
Cordylobia first instar larvae, the spine when at rest is directed
upwards almost at right angles to the cephalo-pharyngeal sclerite,
and when in action its movements are directed upwards to lift the
cuticle. Neither m its shape, nor direction, nor in its action, has it
any resemblance to the mouth hooks of the second and third instars,
which are, according to Lowne (1890), represented in the first instar
larva by the oral rods. This median buccal spine is not present
in the first stage larva of Auchmeromyia, which larva does not .
penetrate the skin; it is significant that it 1s present in Hypoderma
bovis and lineatum, and is figured by Laake (1921), and that it
persists in them up to, and including, the third instar. The
retention in this case is easily understood when we consider the
active migratory character of these stages.

The respiratory system consists of two main tracheal tubes which
commence at the posterior stigmata on the thirteenth segment and
run forward as parallel bilateral tubes; up to the tenth segment they
are yellow in colour; in the region of the eleventh segment there is
a transverse tube connecting them, which is also yellow in colour.
Along their entire course forward they give off branches, and thus

461

they become attenuated anteriorly; no anterior stigmatic opening
can be made out.

Habits of the first instar larva. The larvae remain in the
situation in which they hatch out just below the surface of the sand;
they are difficult to see, and even close inspection of the surface with
a hand-lens may fail to reveal their presence. If, however, the
container in which they are present is vibrated by tapping it, or by
shaking the table on which it stands, or if the surface of the sand
is disturbed by blowing on it or touching it with a needle, the
larvae make their way up rapidly through the few grains of sand
covering them. Similarly they come quickly to the surface and
wave about, if a vessel containing hot water is brought near the
surface of the sand in which they lie concealed. When the larvae
are so disturbed it is easy to observe them with the naked eye;
they adopt a characteristic attitude, being attached by the posterior
end to a grain of sand, the rest of the body being raised in the aur,
and waving about actively as if seeking for something to which to
attach themselves. If any object is allowed to touch the larvae they
at once adhere to it and quickly crawl up on it. Camel’s-hair
brushes were used at first for picking up the larvae, but the larvae
have the habit of at once creeping in between the hairs and
disappearing from view in a few seconds; brushes were, therefore,
discarded in favour of handled needles for the purpose. There is
need for great care, however, that they are not injured in
transferring them from the needle on to any other object such as a
skin surface, as the slightest accidental pressure may render them
very slow in penetrating skin or even incapable of doing so.

Resistance of larvae of the first instar to various agents.

Room temperature. Left in sand at room temperature,
larvae lived without food for about nine days, as a rule; some died
much earlier, and a few lived as many as fifteen days.

On cloth. Larvae two days hatched were taken up on cloth
by laying it gently on the surface of sand containing larvae; the
cloth with larvae adherent to it was kept at room temperature; the
larvae lived on the cloth for nine days; a portion of wet cloth was
used to pick up larvae, and allowed to dry with the larvae on it;
the original condition of the cloth in this respect made no perceptible
difference in the length of time larvae could live on it.

462

Direct sunlight. Larvae on a watch-glass exposed for
twenty hours to open air where the sun reached them during
the whole day did not die, and were capable of penetrating skin.
Larvae in sand exposed to the sun for two hours in the heat of the
day on one occasion only, and thereafter kept at room temperature,
lived for over eleven days.

Dry heat. Larvae were placed in small dry tubes plugged
with cotton wool; the small tube was placed in a test-tube large
enough to contain also a thermometer, and the large tube was placed
in a water bath. A range of temperatures and exposures was tried,
and it was found that rapid definite effects were obtained at a
temperature of 50°C. and above. After two minutes’ exposure at
50° C. the larvae became motionless and failed to recover.

Incubator at 37°C. On dry sand in a watch-glass the
larvae lived for three days-only.

Hot water. In this experiment the small tubes containing
the larvae to be tested were filled with water and plugged, and
placed in a large test-tube half full of water, which was placed in
the water bath. For this series forty-three larvae were used at
various temperatures. [hey survived one minute at temperatures
below 48°C., but one minute at 50°C. and higher temperatures
killed them. ‘Temperatures from 45°C. to 48°C. for two minutes
gave irregular results.

Cold. Larvae placed in a watch-glass and laid on ice, and
kept there for twenty hours, became motionless but did not die; on
removal to room temperature they quickly became active and were
able to penetrate the skin of a guinea-pig.

Immersion. pneq@olid ta pawat eg sap aes aitacuedmre
needles were sunk in tubes of water, and frequently remained
attached to the needle for long periods. Consecutive immersion for
ten and thirty minutes produced no result; larvae immersed for ten
minutes and allowed to dry, and then put in water for thirty
minutes, were active at the end of the time; one made an unsuccessful
effort to penetrate skin. For longer periods this method was
inadequate as the larvae floated up to the surface of the water,
where some of them remained active for three days. Complete
immersion in tubes for long periods—in one instance up to twenty-
four hours—was not fatal to them.

i
Z
E

463

Troning. The process of ironing was fatal to larvae in cloth,
even when covered with several layers.

Phenol. Solutions of phenol of a strength greater than
12 per cent. killed larvae so quickly that they were unable to crawl
out of the solution: solutions of less strength did not prevent them
crawling out. Watch-glasses were discarded in the subsequent
experiments, which were carried out as follows:—A drop of the
reagent to be tested was placed on a slide, the larva placed in it, and
over the larva a small square of filter paper soaked in the reagent
was placed; the filter paper diminished the activity of the larva in
crawling out of the fluid. Ten per cent. phenol and 5 per cent.
phenol killed larvae in five minutes, while 1 per cent. sometimes
failed to kill in ten minutes and frequently failed to do so in five
' minutes.

Sodium hydroxide. Solutions from 20 per cent. down
were tried ; all strengths down to and including 1 per cent. killed in
five minutes.

Formalin. Five per cent. solution killed in all experiments
in ten minutes, but not always in five minutes; 1 per cent. did not
kill in ten minutes, but did in twenty minutes.

Chloroform water in the strength of 40 per cent. chloro-
form killed in ten minutes, but not always in five minutes.

Calomel powder. Larvae placed on calomel powder did
not die, but moved about actively in it for an observation period of
forty-eight hours.

The effect on the first instar larva of oily substance is mentioned
under prophylactic experiments, and the resistance of second and
third instar larva is more properly dealt with under Treatment, as
these stages are already lodged in the tissues of the host.

The skin penetrating power of first instar larvae. Experiments
made with larvae which had hatched from a few hours to as many
as fifteen days previously, showed that they were capable of
penetrating the healthy skin of various living animals as long as
the larwae remained active. Of the fifteen days old larvae tried,
only one penetrated, and that not completely; at twelve days old
many larvae penetrated easily and completely. Numerous experi-
ments were carried out on the skin of man, European and native,
and on chimpanzee, dog, cercopithecus spp., cat, bush cat, guinea-

464

pig, wild rat and fowl; in all these cases penetration of the unbroken
skin was accomplished. Larvae proved unwilling or unable to
penetrate the skin of frog, lizard and python. Where penetration
of the skin was successful, great variation was noticeable in the time
required for the larva to conceal itself under the skin. ‘The animals
in which entry to the skin was effected most expeditiously were very
young wild rats, brown or black, and in these, as in other animals,
the different regions of the body offered differing degrees of
resistance to the boring powers of the larvae. For example, six
larvae penetrated the shaved skin of the rat abdomen in from twenty-
five seconds to one minute; other six placed on the soles of the feet
of the rat required from thirty seconds to two minutes. Again on
the shaved skin of the thorax or abdoinen of the guinea-pig, larvae
penetrated in from thirty seconds to a minute and a half; larvae ~
placed on the soles of the feet required from seven minutes to twenty-
five minutes. Occasionally larvae succeeded only in _ partially
penetrating the skin; but in no case observed, where the larva
succeeded in concealing itself, did the process occupy more than
half an hour.

Method of penetration. When an uninjured larva is
deposited on the skin of an animal which is suitable, the larva
quickly crawls into the nearest groove or wrinkle in the skin, puts
down its head and commences to bore in. The median black mouth
spine can be seen in active movement, the cephalo-pharyngeal
sclerites moving in unison with it; the movement of the spine is
directed to piercing the cuticle and enlarging the aperture on both
sides ; there is a considerable range of lateral movement of the spine,
and a corresponding movement of the sclerites to which it is
articulated. Once the entrance aperture is large enough to admit
the cephalic end of the larva, the body very rapidly insinuates
itself under a thin tunnel of cuticle; the rapidity depending on the
thinness and softness of the skin. The action of the mouth-parts
and the method of using them were studied very carefully in many
experiments; particular attention was given to this, as the result of
the observation that in the dead fixed larva of the first instar the
median buccal spine was usually directed dorsally, and was not
curved down ventrally as are the mouth hooks of the second and
third instars. The experiments were carried out by snipping off a

465

very thin layer of skin from recently killed rats and placing it on a
slide under the microscope and then placing on it a larva at a point
where the action of the mouth-parts could be followed; if the larva
entered at a point not desired, the skin could be manipulated with

Fic. 2. First instar larva penetrating skin. M.G.T.

needles, so as to bring the point of entry into view. In this way
it was seen that the median buccal spine was used for penetrating
the cuticle and then separating it from the subjacent tissues, by a
series of punching movements directed forwards and extending

466

laterally until an entrance aperture was made; then followed a
succession of upward movements of the mouth spine. An illustration
which may serve to explain the upward movements is that they
resemble the upward movements of the points of a tack-lifter, but
in this case, instead of two points, there is only one formed by the
buccal spine. In all cases observed, with one exception—a larva
penetrated perpendicularly into the ear skin of a dead rat—the first
entrance was made into the most superficial layers of the skin in a
horizontal direction: the result was that as the larva lay parallel to
the surface in a thin-roofed tunnel, the black buccal armature could
be discerned through the layer of cuticle covering the larva. It was
interesting to see that on the tail of the rat, the larvae, no matter
in which direction they faced when laid upon the hairs, quickly
turned themselves in such a way that they worked upwards towards
the base of the tail. They followed a hair along to its origin from
the skin and then proceeded to bore in, sometimes near the base of
the hair, but more commonly through the skin between the hairs.
The instinctive action of the larva appears to be to obtain as quickly
as possible shelter and a resting place in the skin superficially,
leaving the further operation of boring in deeply to be done at
leisure, when it is securely ensconced and is safe from the risk of
being rubbed off by the animal. The posterior segment was
invariably left protruding slightly from the aperture, but could be
drawn in when touched.

Penetration ‘of the'skin Of the toatdavcreer xem
ments were made in order to ascertain whether first instar larvae
were capable of penetrating the skin of dead animals. In the first
experiments the animal used was a young specimen of Ratfus rattus
which had been dead six hours. First instar larvae of various ages
were placed on the skin of the ears and tail; in eleven consecutive
trials the larvae eagerly attacked the skin and penetrated completely
and as rapidly as they did in the living animal. In the second
experiments the foot of a dead guinea-pig was used; here again
penetration was accomplished with normal rapidity.

Existence in cadaver. Although in no case did larvae which had
so penetrated complete their development, they proved capable of
living in the tissues of a dead animal for two or three days; on one
occasion a larva which had penetrated a cadaver reached the stage

467

of the first ecdysis. If an animal which had larvae of the second or
third instar present on it died, the larvae as a rule at once migrated
from the dead body and buried themselves in sand or soil; in the
case of one guinea-pig, however, several second and also third stage
larvae did not leave the host, but remained active in the tissues for
twenty-four hours, before they died.

Secondary penetration of skin. Rodhain and Bequaert removed
larvae at an advanced stage from their site in the skin of the host
and inserted them into artificially made cutaneous pockets in a
different host, and observed that they were capable of developing.
Roubaud, as a result of his experiments, concluded that not only
older larvae once removed from the host are incapable of penetrating
skin afresh, but also that young larvae of the first instar are equally
incapable of doing so. This he attributes to a sudden biological
modification of the larva. We are in agreement with this observer
as regards the lack of penetrating power of the second and third
instar larvae, but our observations differ from his in regard to the
first instar. Second and third instar larvae expressed from cavities
in living guinea-pig skin made strong and often successful efforts
to regain the position where they had been lodged; they made
similar efforts to regain their position in tissues of dead animals
on some occasions. They proved, however, quite incapable of
penetrating unbroken skin, either of the same animal or of other
animals. First instar larvae, however, proved capable of penetrating
skin afresh, :f removed from the original site within some hours of
their entrance; early in the instar they are capable of re-penetration
of skin, but later they are not. In Table I are given the details of
some experiments.

From the table it is seen that in three experiments with euinea-
pigs and rats, first instar larvae succeeded in re-penetrating skin
completely ; second and third instar larvae failed to do so. We
consider that the factor which renders it possible for larvae early in
the first instar to re-penetrate, whereas late in the first instar they
are incapable of doing so, is the possession in a functional state of
the median buccal spine. This apparatus is as well adapted for the
purpose of skin penetration, as the mouth hooks of the second and
third instars are ill adapted for the purpose.

The effect of reagents in regard to skin penetration. The

4.68

cadaver of the young rat was used and powders and oily substances
were applied to the skin, and the larvae placed on skin so treated.
It was found that French chalk, borax or calomel, delayed them, but
did not prevent them penetrating the skin. Orly substances,
however, had a great effect, the effect being similar for palm oil,
vaseline and liquid paraffin. ‘The larvae placed on skin treated
with these did not proceed to bore into the skin; they commenced
wandering about, trying to get out of the layer of liquid, and could
do nothing as long as they remained in it. ‘The disadvantages,
however, are obvious, as the film of oily substance must be of
considerable thickness ; it is improbable that on such lines a practical
prophylaxis can be evolved, as when larvae are free from the liquid

they can penetrate the skin, although more slowly.

Taste |

Giving the results of experiments to test the secondary skin-penctrating powers of the larvae
of C. aathropophaga, Grinberg.

or
Instar Animal Animal — Time required
No.’of | No. of | tof{ | from which | on which | for | Remarks
Exp larvae | larvae | removed tested. penetration |
Prenee ER eB | — = — ——————— ee
Min. Max.
I 2 3rd. | Brown rat | Black rat Did not penetrate
|

2 2 3rd_ | Brown rat | Guinea-pig | Did not penetrate

3 2 2nd | Guinea-pig | Guinea-pig | | Did not penetrate

4 I 2nd | Guinea-pig | Black rat | Did not penetrate

5 6 2nd | Guinea-pig | Black rat Did not penetrate

6 I ist | Guinea-pig | Guinea-pig | | Did not penetrate

10 2 Ist | Black rat Black rat Kee 3 min. | Penetrated

Food of first instar larvae.

First instar larvae grow slightly

larger if left in contaminated sand at room temperature, but it is
not possible to say whether this increase of size is due to the
ingestion of food material which may be taken up in small quantities
from the sand. The larvae did not show any capacity for existing

469

long, or developing on such substances as fruit of various kinds,
pieces of muscle, blood or liver of rats. Living tissue affords their
natural food.

Sina A v

t v ‘ 3 ¥v

eG 7%, v vs v
dd tv ” v vy”

yy vov vv. MT SE ae a #

Ge tev, vows yyw ra SS

’ err
PPP Vive ONO giViices oie
yee VNy vy Ad

M.G.T.

Fic. 3. (1) Larva of second instar, ventral view. (2) Larva of third instar, ventral view.

THE SECOND INSTAR LARVAE. This stage arises by a moult
which occurs in the tissues of the host generally about the third day

after penetration; the larva then measures 2°5 to 4 mm.; the

470

appearance and shape of the larva differ entirely from that of the
first instar; whereas the first instar larva 1s somewhat fusiform in
shape, the larva of the second instar 1s club-shaped; it expands
quickly from the cephalic end to about the seventh or eighth
segment, then narrows quickly and is more or less cylindrical to the
posterior extremity. The cuticular spines in this instar are one of
the most striking features, being of large size, black in colour, and
distributed irregularly over the surface of the third to the seventh
segments. The majority of the spines are directed backwards, but
spines directed laterally or even forwards occur rarely; some of the
spines are bifid. The first two segments bear a few rows of brown
spines of smaller size more closely arranged on the ventral than on
the dorsal surface. The segments number eight to thirteen, appear
almost bare in contrast with the foregoing segments; close inspec-
tion, however, reveals the presence of a few rows of small pale spines
on the posterior border of each of these segments up to the twelfth,
which has numerous rows of similar spines on it. The last segment,
number thirteen, which is indistinctly demarcated from number
twelve, is devoid of spines; on its dorsal surface the stigmatic
orifices open. [here are two pairs of processes on the posterior
margin of this segment, the outer pair being larger. The anal
orifice which opens on the ventral aspect of the twelfth segment is
provided with two lateral processes; all these processes are small,
compared with the corresponding processes of the first instar. The
cephalic segment viewed from the dorsal aspect, presents two
rounded eminences, separated from each other by a shallow mesial
sulcus. Each eminence has on its upper surface two structures,
papilla-like; around the base of each is a brownish ring of chitin,
and in the substance of each papilla are what look like delicate
chitinous tubes opening on the surface. The papillae are situated
one posteriorly and the other anteriorly on the rounded eminence,
and each arises in connection with a goblet-shaped structure from
the lower end of which a narrow cord passes backwards. On
the ventral aspect of the segment the tips of the two black buccal
hooks protrude; external to them on either side is a yellow ridge
bearing small spines.

The mouth parts consist of two black hooks strongly curved
ventrally, in contrast with the median spine of the first instar.

ae a

471

Posteriorly the mouth hooks articulate loosely with a hypostomal
sclerite consisting of short rods united by a transverse bar. This
sclerite articulates in turn posteriorly with the pharyngeal sclerites
which pass backwards, reaching the middle of the third segment in
extended specimens.

The respiratory system consists of two main longitudinal
tracheal tubes which pass forwards from the posterior stigmata on
the thirteenth segment to the posterior lateral border of the second
segment, where they end in the anterior stigmata. The posterior
stigmata consist of two curved slits, slightly oblique in direction, the
concavities of the curves facing each other. [he main tracheal tubes
are chitinized to about the middle of the eleventh segment; just
anterior to the chitinized portion a transverse tube connects the two
lateral tubes. In the region of the third segment the main tubes,
which have given off branches all the way forward, suddenly narrow
and continue as a fine chitinized tube to the anterior stigmata, each
of which consists of a fringe of finger-like processes about eight in
number. Great variation in size 1s seen in late second instar and
early third instar larvae even from the same host.

THIRD INSTAR LARVAE. The second ecdysis occurs in the host
tissues from the fifth to the sixth days, and the resulting larva
matures and leaves the host about the eighth day. The extended
larva measured, when mature in rats, from 13 to 15 mm. long. Not
only the measurements, but also the date on which the larva reaches
maturity and the date on which the ecdysis occurs is influenced by
the relative suitability of the host. The mature larva consists of
twelve segments; it is roughly cylindrical in shape, its greatest width
occurring at the seventh and eighth segments. ‘The first segment
is the smallest, and is frequently retracted into the second. It
presents dorsally on each side a protuberance carrying two
segmented papillae, the basal segment being chitinized. Ventrally
on the segment are seen the free extremities of the two black buccal
hooks sharply pointed and curved ventrally; on either side of the
hooks there is a ridge of yellow chitinized integument bearing a row
of small spines, about six in number. The mouth hooks articulate
posteriorly with the hypostomal sclerite, which in turn articulates
posteriorly with the pharyngeal sclerites; the posterior end of these
reaches to the junction between the second and third segments. On

472

the second segment at its posterior margin laterally, the anterior
stigmata open; these consist of a fringe of finger-like processes about
ten in number. The twelfth segment is small; near the anterior
margin of its dorsal surface the posterior stigmata open close
together on rounded eminences; they consist of three sinuous slits
obliquely placed; internal to the slits is a small circular opening ;
there does not exist in this case a definite chitinized plate on which
the stigmatic slits open. The cuticle of segments number four to
eleven, on the ventral surface, 1s thrown into folds of an irregular
appearance, which are more pronounced in the middle segments,
and are concerned: in locomotion. Backwardly directed curved
spines are present on segments two to nine, being more numerous
and of darker colour on the sixth, seventh and eighth segments; on
the ninth segment few spines are present, on the tenth still fewer,
while the eleventh and twelfth are practically bare. The white
appearance of the last four segments forms a marked contrast with
the speckled black appearance of the anterior segments. The great
diversity of appearance which the larva of Cordylobia presents in
its different instars and at different stages of the same instar, has
induced some observers to introduce separate names for them; this 1s
of little assistance, and tends to add to the already sufficiently great
confusion which exists in regard to the larvae of flies causing
Myiasis.

(4) PUPARIUM

PUPATION. The process of pupation was observed in larvae
which had been removed either from naturally infected or from
experimentally infected animals. If mature the larvae commence to
pupate within twenty-four hours; the anterior extremity becoming
pinkish at first, then terra-cotta coloured; the colour extends along
the body to the posterior end, and later darkens to a dark chestnut.
If the larvae are immature, pupation may be delayed for a day or
two, or the larvae may die and turn dark in colour; if the larva has
been chloroformed or has been removed from a sloughing septic
cavity, it becomes rapidly black from before backwards, the
puparium fails to separate and harden, and the larva dies. The
shape of the puparium is rather characteristic. It has the posterior
end very squarely cut off and the sides run parallel to each other,
giving an elongate oblong appearance; it tapers somewhat abruptly

473

D1. Get

Fic. 4. (1) Posterior view of first instar larva showing processes. (2) Cephalic end of
second instar larva. (3) Anterior stigmata of second instar larva. (4) Posterior stigmata of
second instar larva. (5) Cephalo-pharyngeal skeleton of third instar larva. (6) Posterior
stigmata of third instar larva. (7) Puparium.

474

at the anterior end. On the anterior extremity of the puparium are
visible two small papillae, and the posterior stigmata can be
discerned in most specimens. In all cases the rings corresponding
to the segments are seen, and the well defined spines of the larvae are
easily made out. When the puparium is treated with phenol in
order to clear it, the hard case is still more easily identified as the
last larval moult, the mouth hooks and attached sclerites, the
posterior tracheal tubes and stigmata, as well as the cuticular spines
being readily demonstrable under the microscope.

MEASUREMENTS OF THE PUPARIUM. The smallest obtained
measured 6°5 mm.; it was derived from a rat which died before the
larvae were mature; the largest obtained was 11°5 mm. long; it was
derived from an experimentally infected wild rat in which the larvae
matured. Below is shown the distribution according to length of
a series of forty-three puparia derived from the naturally infected
wild rat mentioned above.

& an|hropophaga ; from a. qalurally intecled ral which died.

Grapn I. Showing the distribution according to length
of 43 puparia of Cordylobia anthropophaga, Grinberg.

475

SITE CHOSEN FOR PUPATION.. Larvae which had been removed
from the host, or had left the host, were observed; when placed on
sand which was dry they began after a short time to dig down into
it, and in most cases in an hour or two they were out of sight.
Sand was then provided in small tubes for each larva; the sand
was so arranged that there was a damp layer of sand at the bottom
of the tube of one inch in depth, and a dry layer of sand above this
half an inch in depth; over fifty larvae were tested, and it was
found that with a few exceptions all went right to the bottom of the
tubes to pupate; the exceptions reached only the top of the damp
layer. Occasionally pupation occurred on a dressing applied to an
animal’s limb over a tumbu lesion.

EXPERIMENTS WITH PUPARIA.

At room temperature. On 16th February, 1923, nineteen
puparia derived from the larvae of a wild rat were placed at room
temperature; of these seventeen emerged either on 26th or 27th
February, 1923. One pupa failed to give rise to an adult, and one
adult died while emerging backwards; it may be noted that on
several occasions the fly was thus inverted in the puparium.
Newstead (1907) drew attention to this occurrence in the case of
Auchmeromyia luteola.

In the ice chest. On 16th February, 1923, twelve pupae from
the same source were placed in an ice chest. None of these had
emerged by oth March, 1923; they were then placed at room
temperature, and of the twelve, nine emerged on 14th March, 1923;
the other three failed to emerge by ioth April, 1923, when the
experiment was terminated. The resistance of the fly in the pupal
stage to cold is of-interest, as is also the marked prolongation of
the pupal stage under these circumstances.

In Incubator at 37° C. On 16th February, 1923, twelve pupae
from the same source were placed in the incubator at 37°C.; on
2nd March, 1923, none had emerged; eight were removed from the
incubator and placed at room temperature, four being left in the
incubator. Of the eight removed, four were kept dry and four were
kept moist. On oth March, 1923, none of the twelve had emerged ;
the four remaining in the incubator were placed at room tempera-
ture, dry. On roth April, 1923, none of the twelve had emerged
and the experiment was terminated. ‘The resistance of the fly in

476

the pupal stage to dry heat is therefore small. A similar observa-
tion has been made by Dove (1918) on the resistance to heat of the
pupae of Gastrophilus haemorrhoidalis, |

Temperature and dryness. \n order to determine, if possible,
whether the failure to emerge at 37°C. was due to the effect of
temperature or dryness on the pupae, the following experiment was
carried out with six pupae derived from a natural infection in a
mongoose. Three pupae which had pupated on 19th and 2oth
March, 1923, were placed at room temperature in a calcium
chloride desiccator. Three adults emerged on Ist April, 1923, from
the three pupae.

Three pupae which had pupated on 21st March, 1923, were
placed in the incubator at 37°C., on 11th April, 1923, none had
emerged ; they were taken out and placed at room temperature; on
16th April, 1923, none had emerged and the experiment was
stopped. So far as these experiments go, it appears that it is the
height of the temperature and not the lack of moisture which
prevents the development of Cordylobia at 37°C.

METHOD OF EXIT OF FLY. The puparium is broken at a short
distance from the anterior end, and the whole cap or segments of it
come off. The fly on emerging pushes vigorously through any
resistant substance to escape; it was interesting to see that where
puparia had been placed on a plug of woo! half-way down a test-
tube, plugged also at the mouth with wool, the flies which emerged
most frequently went downwards through the wool on which the
puparium was lying so that finally they arrived at the bottom of
the test-tube. The newly emerged fly possesses great powers of
forcing its way through such substances as sand, and cotton wool.
As soon as it emerges, it sets about the operation of getting out of
its immediate environment. The pressure exerted by the fly was
very remarkable; in many cases two inches length of tight cotton-
wool plug in the mouth of a test-tube being insufficient to prevent
them escaping; in a few instances where the plugs were used, flies
had flattened themselves between the plug and the glass to such an
extent that they died, with the ptilinum extended in front of them
and the wings still unexpanded. Dove (1918) carried out some
experiments with Gastrophilus haemorrhoidalis puparia, which also
showed the capability of the adults to push their way through

477

obstructing material. Thirty-two puparia were placed in moist
loam at a depth of five inches; of these twenty-nine emerged
normally, the adults pushing their way up to the surface.

Ecpysis. ‘The insect from the time it hatches out of the egg
as a larva until it emerges from the puparium as an adult fly has
undergone four ecdyses. ‘The larva undergoes the first and second
ecdyses in the tissues of the host, the first occurring from the second
to the fourth days, and the second from the fifth to the sixth day.
These dates were determined by removing larvae from their site in
the skin of experimental animals at regular intervals after their first
entrance, and examining them. The skin is cast from before
backwards; the new stage emerging from the anterior end; we have
been fortunate in finding larvae at various stages of the moulting
process ; for example, some specimens showed a condition in which

M.G.T.

Fic. 5. (1) First ecdysis, cephalic end. (2) First ecdysis, caudal end. (3) Second
ecdysis, caudal end.

successive stages of the buccal armature were present together, while
the posterior stigmata were still only in the earlier stage. Other
specimens had the anterior portion of the cast skin loosened and
crumpled backwards, while the posterior extremity of the cast was
still firmly adherent, and showed the posterior stigmata of the
successive stages co-existing (see fig. 5).

The third ecdysis occurs when the fly pupates, the larval skin
not being got rid of, but remaining to form the puparium; the pupa
which arises inside the puparium undergoes an ecdysis; this
constitutes the fourth ecdysis. The pupal sheath cast off is found
in the posterior end of the puparium when the fly has emerged.

478

Pupae removed from the puparium a little time before the time for
the fly emerging were found to be enveloped sometimes completely
and sometimes partially in the pupal sheath. In some cases it
remained only as a delicate covering on the legs.

SIZE OF ADULTS. In the experiments related above, where
certain pupae were subjected to the temperature of an ice chest, and
in which the pupal period was thereby prolonged greatly, the size
of the emerging flies was not appreciably affected. Of the flies which
emerged in the experiment, twenty-two were measured: the males,
ten in number, measured from 8 to 10°5 mm.; the females, twelve in
number, measured from 8°5 to 10°5 mm. It is noteworthy that
the size of the larva is no accurate guide to the size of the
resulting puparium nor of the emerging adult; it is only a guide
within rough limits. For example, two larvae gave the following

figures :—
Size of larva
—__——— Size of Size of adult
Contracted Extended puparium
Lye eae “7 II°S g°0 8°5
2. hee a Rae 9°5 I1"o I0°0 10°5

The size of the emerging fly depended, however, on factors of
which the degree of development of the larva is clearly one. Larvae
which are removed early and which pupate give rise to flies which
are small, while larvae which mature and pupate naturally give
rise to large flies. In Plate XVI are shown two series, the first
males and the second females, which demonstrate clearly the great
variations in size which the fly may present.

FERTILISATION OF THE FEMALE. Copulation took place at once
when recently emerged females were placed with wild male flies;
it did not matter whether the females had fed or not; copulation
lasted about two minutes and was repeated several times during
the first day: after that it was not seen to occur; newly emerged
males did not copulate readily.

479

V. DEVELOPMENT OF C. ANTHROPOPHAGA Grun. IN ANIMALS

(1) DURATION IN EXPERIMENTS

In the table given below are shown the various stages of the fly
as 1t occurred in two of the experimental animals, a guinea-pig and
a wild rat.

It is seen from the table that the development times in these
cases were nearly the same, but the regularity with which the larvae
in the rat completed the development was greater. As a rule, we
found also that adults from rat puparia, as well as the puparia,
were on the average somewhat larger than those from guinea-pigs.

(2) PATHOGENICITY TO ANIMALS

IN NATURE. The animals most commonly found by us to be
affected were wild rats, both brown_and black; the feet, the genitals,
the tail and the axillary region were chiefly involved where single
larvae were present; in heavily infected animals any site was
apparently suitable, including the nose. Dogs were also affected,
and other animals more rarely. Lesions of the feet of sheep and
goats were attributed by natives to the larva, but we did not find
evidence of infection in those examined—about fifty. Various
animals presented old scars and also suppurating sinuses which
from the appearance might have contained larvae. The resulting
lesions produced by the larvae during growth are illustrated by the
case of a wild rat which had torn part of the skin off its abdomen
in endeavouring to get rid of the larvae. As a rule, however,
animals even with many larvae present either would not, or could
not, get rid of them even when in quite accessible places. Several
rats and a mongoose died apparently as the result of natural
infection.

IN EXPERIMENTS. Guinea-pigs which were allowed to walk on
infected sand soon showed signs of infection in the papule forma-
tion on the feet; by the third day the feet began to swell and the
animals were seen biting them; in two days more great oedema of
the feet was present, and in the oedematous skin the larvae could
be seen; the posterior end was below the surface as a rule, and the
circular aperture was smooth and polished on the margin; the

480

Taste II.

Giving the development of C. anthropophaga in Guinea-pig and Wild Rat.

Animal

Guinea-pig

R. rattus...

Larva
Number

Days in
skin

Days to
pupation

Days to
emergence
of adult

Remarks
Removed for experiment

Did not pupate.

Did not emerge.

Larva lost.

———$

Pupa preserved.

481

posterior stigmata: are easily seen with a hand-lens at this stage,
and also the more striking tracheal tubes leading forward from them,
which appear as two parallel silvery lines. The cavities occupied
by the larvae are of some depth, because often the posterior end of
the larvae is not flush with the skin surface, but lies several
mulimetres below this; as the mature larva may measure up to about
15 mm., it 1s seen that the larva may reach with its mouth parts a
point about 2 cms. from the skin surface. One guinea-pig in this
experiment died as the result of the infection; the larvae were found
to have penetrated to the tendons of the feet; the overlying tissues
were very greatly thickened owing to the oedema. In one guinea-
pig where a larva had attacked the abdominal skin, it had
penetrated so deeply as to cause a tumour pushing inwards the
parietal peritoneum, which was congested deeply and _ thickened.
Wild rats and small animals frequently died as the result of
infection.

In sections cut through the larva in situ in the skin of animals,
where no sepsis was present there was noted only oedematous
thickening and moderate round-cell infiltration of the tissue
surrounding the larval body.

(3) THE ANIMAL HOSTS IN NATURE OF C. ANTHROPOPHAGA Grin.

From various sources, beginning at the early observations of
Coquerel and Mondieére (1862) and coming down to the present day,
we have collected a list of animals in which larvae of Cordylobia
have been found. In the notes of most observers the two animals
which come first are man and dog, the other hosts appearing in the
records of a more limited number of observers. Neave (1912-13)
mentions that dogs suffer badly, and a case ina rat. It is a very
natural thing that especial note should be made of conditions_
affecting man, and also that any condition affecting the dog should
be the subject of remark, owing to the intimate association of this
animal with man. It is this natural tendency which explains, we
believe, the commonly held opinion that the dog is the usual host
in nature of Cordylobia; of this belief we shall say something later.
We have found the following animals recorded as hosts :—Man,
dog, guinea-pig imported and locally bred, wild rat, various

482

monkeys, white rat, cat, wild cat, arvicanthis, squirrel, goat and
antelope. Roubaud (1914) expresses doubts about the occurrence
in goats and antelopes. A case of a mule infection was reported
to us, but we had not the opportunity of investigating it. We have
found larvae in addition in the mongoose and chimpanzee at

Freetown.
(4) THE MAIN NATURAL RESERVOIR OF THE INFECTION

It is a matter of great importance to determine which animal
forms in nature the reservoir in which the infection is maintained
and from which man derives his infection. Le Dantec and Boye
(1904) wrote, ‘Le chien est l’animal de choix pour la culture de la
larve.’ Roubaud was struck by the fact that dogs are so often
affected, as reported by previous workers and also from his own
observations. He writes, of dogs, ‘Ce sont ces animaux qul
constituent le reservoir permanent de la myiase furunculeuse,’ and
that there is a striking relationship between the human cases and
the presence of dogs in the immediate vicinity of those men who are
infected. More than this, he concludes that the rarity or absence
of dogs in any region is one of the most immediate causes of the
absence of the larva from that region. ‘ Il semble que l’abondance du
ver dans un pays soit souvent fonction de celle de la population
canine.” The logical deduction which Roubaud makes is in regard
to the prophylaxis of Cordylobia Myiasis; as dogs are the natural
reservoir, it 1s against the infection which exists in dogs that man
must take action. The action recommended by him consists in the
regular inspection of dogs at least once a week, the removal of
larvae from them, and the destruction of the larvae. He anticipates
considerable results from carrying out this procedure, ‘ En détruisant
ainsi quantité de parasites, a la source méme qui les entretient
normalement, on arrivera nécessairement a faire disparaitre la
Cordylobia des endroits qu’elle infeste, ou tout au moins a la rendre
extrémement rare.’ There is no doubt but that a careful attention
to the expression and destruction of larvae in dogs will diminish
the numbers of Cordylobia, and here we are in complete agree-
ment with Roubaud. Where we differ is in regard to the question
of dogs being the natural reservoir of the fly ; we do not believe that
the dog, easily and heavily infected as it doubtless often is, plays
such an important part in the maintenance and spread of infection

483

as to justify the optimism of Roubaud with regard to the results of
his suggested method of prophylaxis. We found that in experi-
mental trials, wild rats proved themselves more suitable hosts for
the development of larvae than did even young dogs, that in nature
wild rats were more frequently and severely infected than dogs, and
finally we have been able to prove a close association between
Cordylobia and wild rats, by finding in burrows of rats puparia of
Cordylobia and rearing from them adults.

In one comparative experiment, using two pups and _ two
medium-sized wild rats, twelve first instar larvae were allowed to
penetrate the skin of the abdomen of each animal. Two larvae
developed on pup number one, three on pup number two; on rat
number one, eleven developed, and on rat number two, eight. The
larvae from the dogs left their hosts on the tenth day, those from the
rats left their hests on the eighth day; the average size of the dog
larvae was 12 mm. contracted and 14 mm. when extended, of the rat
larvae was 12°5 mm. to 14°5 mm.

In nature, apart from large numbers of rats which had a few
larvae in their tissues, we observed cases in which very severe
infestation caused death. For example in a wild rat, R. vattus,
nearly full grown, which was brought into the laboratory in a
moribund state, there were present in various parts of its body forty-
three larvae. The nose, the feet, the genitals, the tail and the
general body surface were affected; the animal was in a very septic
state and died svon after being brought in. Another rat presented
no less than forty-one larvae, and here again the infestation resulted
in death.

We have already seen that first instar larvae penetrate the skin
of young rats more rapidly than that of any other animal tried; the
larvae maintain themselves and develop in a higher percentage in
rats than in dogs or guinea-pigs or any other animal used; the time
taken by the larvae to develop in the tissues is shorter in rats than
in dogs; the larvae which result from the rats are larger than from
dogs. So far, then, as experiments are concerned, there is proof
that the rat is a more suitable animal host for the larva than is the
dog. In nature also we have found the rat more suitable.

We believe that Roubaud himself was aware of this high
susceptibility of wild rats from the following three facts. The first

484

is, that he knew of the observation of Koch in East Africa reported
by Donitz; in East Africa, Koch found a disease in epizootic form
which was killing wild rats; this disease on investigation proved
not to be due to plague bacilli as first suspected, but to fly larvae
present in the tissues; it was, indeed, this fly which Donitz named
Cordylobia murium, and which Roubaud is at some pains to prove
is, in reality, C. anthropophaga affecting wild rats. The second
fact 1s, that in discussing certain observations of Delanoe on
Cordylobia in rodents, Roubaud says, ‘ Elles montrent que ce
parasite peut trouver, en l’absence de Vhomme ou de _ gros
mammiuferes domestiques, son réservoir naturel chez de_ petits
rongeurs, avec une électivité tres marquée pour certaines espéces
a l’exclusion des autres.’ The third is, that he made the following
experiments :—-On page 141 of his work, his experiment B on
development in the dog, resulted in twenty-two out of thirty larvae
developing on the skin of a dog; he concludes, ‘ Le chien apparait
donc comme un héte de choix pour l’évolution du ver du Cayor:
c’est la confirmation de ce que l’on constate par l’observation
naturelle.’ There follows on this immediately, however, his
experiment C, on development in the rat; in this twelve larvae
developed out of twelve tried on the grey rat (Mus microdon), i.e.,
four larvae on each of three rats. Of this he says, ‘ La facilité
avec laquelle le ver du Cayor evolue chez le rat, fait de cet animal
Vhéte qui s’indique le mieux pour étude du parasite.’ It is
remarkable that in spite of the evidence which Roubaud himself
produced in the experiments given above, he should have laid such
undue emphasis on the part plaved by dogs in this disease in

nature.

(5) TEMPERATURE OF ANIMAL HOST IN RELATION TO INFECTION

Roubaud (1914) observed that in certain species of vertebrates
there was a more or less complete immunity towards the larvae. He
considers that the development of the larvae 1s easy in proportion
as the temperature of the animal host is low. He gives the following
table of rectal temperatures 1n support of his thesis : —

Rat sie te ph si byte (er
Dog ee. e, nia ‘e ABBR SaR
Guinea-pig ai ck seh dacs 3995
Pig “a a a. nek ite ae

HOWL oe, me +e te KRY. WH.

485

He found as we have seen that, 1n experimental work, the rat
was a favourable animal, and that the fowl was useless for the
development of larvae; we saw that he regards the dog as the
natural reservoir of the infection. Man, however, he regards as
secondary , ‘L’>homme ne _ représente certainement qu’un hdte
accidental, chez lequel l’évolution ne se fait pas toujours.’ This
statement does not accord with the theory that body temperature
plays a part of paramount importance in this matter, as the rectal
temperature of man, 37'2, is intermediate between that of the rat
and that of the dog; from the point of view of rectal temperature
man should form a very favourable host. Moreover, our experi-
ments with guinea-pigs showed that they could often be readily
and severely infected, and that the infection might even be fatal.

(6) DISCOVERY OF PUPARIA OF CORDYLOBIA IN NATURE

As a result of our laboratory experiments and the observations
made on rats infected in nature, we concluded that rats are of great
significance in the maintenance of the infection; the larvae found in
rats must, we thought, emerge to pupate in the places where rats
rested; 1t was decided, therefore, to investigate rat burrows and
search for puparia. Although rats are common in Freetown, it
proved no easy task to find rat burrows which were well defined,
and which at the same time were situated so as to be capable of
being dug out. .Dr. W. Allan, W.A.M.S., the Medical Officer of
Health for Freetown, kindly rendered us great assistance in this
matter, and indicated two accessible rat burrows, one on open
ground; the first burrow yielded puparia of Cordylobia; the soil at
the entrance was carefully searched before digging was begun; the
puparia were found in the first foot of the burrow proper, where the
soil was light, dry and friable; from nine of these puparia which
were still unopened, adult Cordylobia emerged in the laboratory.
The second burrow was in the side of a laterite wall, and one
puparium was found here, but the burrow could not be followed into
the wall. In addition, puparia were found near the bungalow in
rat holes among the rocks, one in each of two localities; from these
adults emerged. Emphasis should be placed on the fact that along
the river front at Freetown, and also along the small streams
through the town, the occurrence of natural hiding places among

486

loose boulders, clefts in the rocks and unpointed walls, makes it
possible for rats to exist easily without the necessity of making
definite burrows. This fact will make the thorough survey of rat
holes for Cordylobia a difficult operation, and will also naturally
render any action against them prohibitive in cost. We are of
opinion that these observations not only add one to the already long
list of diseases for which the wild rat may be responsible, but also
demonstrate that the prophylactic measures against dog infection
suggested by Roubaud are unlikely to be as effective in eliminating
either the fly or the disease caused by it as he anticipates.

VI. THE AGE INCIDENCE OF THE DISEASE IN NATURALLY
INFECTED ANIMALS

It is common knowledge among natives of Sierra Leone that
young children are more often affected by Myiasis due to Cordylobia
than are grown persons; it 1s also known by them that young dogs
are more frequently and more heavily infested than large dogs.
They are so well aware of the fatal consequences which attend
infection with the larvae of young pups, that in some places it is
regarded as best to drown young pups when heavily infested.
Maberley (1918) refers to Dr. K. K. Cross’ observation, recorded in
Sir Harry Johnston’s book on British Central Africa, of maggots
in native children—the whole side of a child riddled with holes.
Marshall (1902) says that in Salisbury one baby had no less than
sixty maggots extracted from it, and that there had been several
cases in which babies had had a dozen or more. Griinberg (1903),
quotes the observation of Steudel, who in Bagamoyo found larvae
regularly on young dogs, but not on grown dogs; in these cases
Steudel thinks that the. fly seems to like to lay its young on the still
soft and moist skin of the new born animal. Le Dantec and Boyé
(1904) say ‘ young dogs above all are severely attacked, and one has
sometimes to remove five or six larvae daily for several weeks in
succession.’ Smith (1908) says ‘ babies at breast and carried in a
cloth on their mother’s back are often affected. . . . Small pups
suffer more than adult dogs and the larvae are all over them.’
Roubaud (1914) mentions that he and Bouet observed numerous
cases among dogs in Dahomey, and that at Khombole, even in the

487

dry season, he found young dogs infested; he adds ‘ Dans le Baol,
jai observé de véritables nids de larves chez les petits chiens. Les
adultes sont d’ordinaire beaucoup moins infestés.’

NON-DEVELOPMENT OF LARVAE IN SKIN OF LIVING ANIMALS

Although uninjured first instar larvae penetrate with great
certainty into the skin of many species of living animal, even when
the larvae are many days old, it does not follow that once
established in the host tissues they will develop to maturity. This
is a matter of considerable significance, and a few detailed observa-
tions may be given.

EXPERIMENTS ON HUMAN SKIN.

Adult European. 30th January, 1923. A larva o'9 mm. long
was placed on the back of the first phalanx of the little finger at
11.30 a.m.; the larva moved quickly into a wrinkle and it proceeded
at once to penetrate, the mouth parts moving very rapidly making
an entrance wound ; the body pushed quickly into the aperture with a
rippling movement, starting from about the ninth segment; no
sensation whatever felt. 11.40 a.m.: First sensation felt when
body of larva half concealed. 11.45 a.m.: Mouth parts ceased
working for a minute; at this time only two posterior segments
uncovered ; the median mouth spine and sclerites plainly visible
through a thin covering of cuticle. 11.50 a.m.: Whole larva
concealed in tunnel of cuticle. 1.30 p.m.: Definite constant itching ;
redness around larva and swelling at situation of anterior end.

31st January, 1923. Definite papule formed; no irritation.

Ist February, 1923. Papule larger; no itching, no irritation.

The after history of this larva, as also of one which penetrated
the same day on the dorsum of the second finger first phalanx, close
to a hair follicle, was that the papule gradually disappeared
without further itching and discomfort. Similar experiments were
subsequently performed on Europeans.

Adult Africans (Young). In two Africans, experiments were
done with four larvae on the inner side of the upper arm.

The history of all these human experiments was similar; the
larvae penetrated rapidly and caused irritation and papule forma-
tion. After three days they caused no more trouble and the

488

papules gradually disappeared. Roubaud applied two first instar
larvae to his arm, and one to that of Dr. Bouet. ‘The three larvae
penetrated normally; as a result there was a partial development
only, which went on for twenty-four hours. After this period no
development occurred and the small lesions healed easily.

EXPERIMENTS ON ANIMAL SKIN.

Guinea-pigs. Eighteen larvae penetrated the skin of the feet
of two guinea-pigs; of the eighteen only eight developed ; these were
removed on the eighth day.

Chimpanzee. Five larvae penetrated the skin of the forearm;
none developed; in this case the animal was not prevented from.
scratching.

Cercopithecus callitrichus. Three larvae penetrated the skin of
the tail, but none developed.

From these experiments the information was obtained that
larvae which have succeeded in penetrating skin do not always
develop; and that this lack of development could not be attributed
in most cases to mechanical interference on the part of the animal.

VII. RELATIVE IMMUNITY OF OLDER ANIMALS

Numerous observers, as we have seen, have recognised the fact
that young animals are more highly susceptible to Cordylobia
infection than old animals. From what we know of the bionomics
of the fly and the method by which first instar larvae gain access
to the animal body, it is improbable that the comparative rarity of
the disease among older animals is due to lack of opportunity of
acquiring the infection. It appears clear that the failure of adult
animals to show infection is due not so much to non-penetration of
the larvae into their skin, as to non-development of larvae after
entrance to the skin. The body is capable of resisting the process
of development of the larvae but not the entrance of the larvae; the
condition is one of relative immunity. If this relative immunity
exists, and we have no doubt from observation and experiment that
it does exist, we must endeavour to ascertain its nature. It is not
an inborn hereditary immunity, because native children and the
offspring of animals belonging to the country do not possess this

489

immunity. It is possible to suppose that it is an immunity which
results from age alone, a form of immunity which it is easier to
postulate than to prove. Evidence against its being an immunity
which arises simply as a result of the maturity of the animal, 1s
provided not only by the first, but by many subsequent observers.
Coquerel and Mondiére (1862) mention the case of an adult spaniel
which was infested by a hundred larvae and died after some days;
Bérenger-Féraud (1872) gives an instance of a spaniel which had
seventy-eight larvae, while a pup of the same breed had three
hundred, and died. Again, adult immigrants to West Africa,
involving many nationalities, including Europeans and, as
mentioned by Blenkinsop (1908), West Indian troops, are affected
by the larvae. It appears improbable that age alone confers
immunity. There remains immunity acquired as the result of
previous attacks of the larvae, and this we believe to be the true
cause of the fact that older animals are less frequently infected than
are young ones. Experimental evidence of such an acquired
immunity is naturally not easy to produce, as the animals with
which experiments are carried out in Tropical Africa are almost
certainly partially immune; the repeated entry of larvae to the
skin and their subsequent destruction by scratching and rubbing
must constantly go on. ‘The following human observations may,
however, be quoted. An adult European in Sierra Leone suffered
from a natural infection, nine larvae developing in the skin; there-
after he proved resistant to infection at several attempts, as shown

below :—
Taste IIT.
Showing the result of attempts to infect a human adult with Cordylobia larvae.
Number of larvae which

Date penetrated skin Site in body Result

September, 1922 ...| Natural infection g | Larvae in upper arm | Removed at
6-8 mm. long

490

Certain animals gave suggestive results. Two dogs which had
been infected on the abdomen with partial success, resisted infection
at a subsequent date with eighteen larvae each, all of which
penetrated. Into the skin of two guinea-pigs which had recovered
from an experimental infection there penetrated on 7th March, 1923,
four and five larvae respectively ; on 15th March, 1923, six larvae;
on 20th March, 1923, six larvae; on 23rd March, 1923, six larvae;
on 28th March, 1923, four larvae. Although the larvae developed
for a period varying from a few hours to two days, in no case did
they develop beyond the first instar. A monkey previously
naturally infected on the root of the tail received on the perineum
on 2nd April, 1923, nine larvae; on 8th April, 1923, six larvae; on
10th April, 1923, six larvae; no development occurred beyond a
papule formation which resolved on or before the third day. These
instances appear to show that there is an acquired immunity against
C. anthropophaga.

On the other hand, we encountered certain paradoxical results,
as for example, the following:—On 7th March, 1923, a Creole
youth received four larvae in the upper arm; development
to papules only; on 8th April, 1923, a single larva penetrated
and this developed normally. Again, a small dog which had
had infection and had thereafter proved resistant to several
infections, at the third attempt received ten larvae on 1oth April,
1923; of these three developed normally. In each of these two
cases anthelminthics had been administered, to the human case
betanaphthol before the last infection, and to the dog carbon tetra-
chloride on the same day as the infection; but whether this fact is
merely coincidence it is not at present possible to say.

Against the immunity theory we have also the fact that large
rats often present infection; we must conclude from this, either that
such rats had not acquired immunity in their early days, or else
that their immunity was broken down by some cause late in life.
It is not possible, with our present knowledge, to explain why this
immunity, as do all forms of immunity, breaks down on some
occasions. The broad conclusion, nevertheless, is that there exists
among adult animals an immunity against Cordylobia larvae, and
we believe that the foregoing experiments point to its being an
immunity acquired through previous attacks of the larvae, whether

491

these developed or were destroyed before developing by mechanical
interference on the part of the host.

DURATION OF IMMUNITY

So far as we are aware, there are few facts available as yet upon
this point; the following observation of Heckenroth and Blanchard
(1913) has some significance and deserves mention. A European
in the Congo had a fox terrier which acquired infection in October,
1911; the owner got infected in November, 1911. The owner and
the dog left Africa and returned in over a year. In January, 1913,
the dog became infected, and in February, 1913, the owner became
infected ; the immunity if established by the first attack, did not
persist for much over a year at the most.

CUTANEOUS REACTION

In the human case, where repeated attempts were made at
infection, there was a severe local reaction at each of the last six
attempts; the penetration of the larvae was accompanied by great
itching and was followed within a few minutes by remarkable local
signs. At the point of entrance of each larva a white bleb formed
and spread rapidly in all directions; the larvae were placed on an
area of about two inches by three, and vet in all cases within fifty
minutes the white urticarial wheals had coalesced and produced an
irregular swelling about three inches in diameter and raised in the
centre about a quarter of an inch; around the white area was a zone
of deep congestion which faded away at the margins. The rather
tense white swelling stood out in a striking manner against the red
background; very considerable itching was felt over the whole area
affected, while at the same time a slight pricking sensation caused
by the larvae boring was experienced. By next day the swelling,
redness, and to a great extent the itching, had subsided.

Hadwen and Bruce (1917) made observations on Anaphylaxis,
in cattle and sheep, produced by the larvae of Hypoderma bovis,
H. lineatum, and Oestrus ovis. Intravenous injection of larval
extracts in saline solution produced death rapidly in some animals,
severe reactions in others, while in some it produced no ill-effect.
The preliminary sensitisation was not experimental but natural, and

492

was attributed to the excretions of the larvae; in one case there was
evidence of the sensitiveness being inherited. Anaphylaxis was
only found to occur when larvae were broken in an animal, or when
a dose had been injected. An ocular reaction occurred in sensitive
animals when a drop of juice from a larva was placed on the
conjunctiva. The cutaneous reaction in our case, while apparently
anaphylactic in nature, resulted not as in Hadwen and Bruce’s
experiments from the injection of extracts, but from natural penetra-
tion of larvae into the skin, the larvae being uninjured.

These observations which we have made on immunity appear
to us of great importance, not only in so far as concerns Cordylobia
Myiasis, but still more in relation to the Mylasis caused
by Dermatobia and Hypoderma, in which cases enormous loss is
suffered year after year on account of damage to hides. The
development of some method of immunizing cattle against the
attacks of the larvae would result in a great increase of value of
hides from countries which are affected by such larvae. The
remarkable thing is that in South America, in the midst of
a country in which cattle are severely affected by the larvae of
Dermatobia, there actually exists a breed of cattle—Antioquia—
which enjoys a relative immunity. We are indebted to Mr. M. T.
Dawe, Commissioner of Lands and Forests in Sierra Leone, for
showing us his photographs of this breed and for much interesting
information concerning it. We feel convinced that a further study
of this subject, with a view to discovering some means of artificially
producing a definite immunity would well repay the trouble and

expense involved.

VIII. SEASONAL INCIDENCE OF C, ANTHROPOPHAGA Grun.

The consensus of opinion of previous observers appears to be
that the wet season is the season of prevalence of infection with
Cordylobia larvae. Coquerel and Mondieére (1862) stated that in
the month of July, in Senegal, after the commencement of the rains,
many cases of these larval parasites occurred. Le Dantec and Boyé
(1904) say that in French Guinea, the adult appears in the
beginning of the wet season, disappears abruptly in October, only
to reappear next year with the first rains. Rodhain and Bequaert

493

(1913) mention a series of animals which are affected with larvae
at Katanga, all in the wet season. Roubaud (1914) refers to
Bérenger-Féraud’s observation that in human beings this form of
Myiasis occurs in July. Howard (1912-13) says that the larva is
very abundant during certain seasons in the Transvaal. The
infection of human beings is considered by Roubaud to result from
close association of dogs with men, the fly being primarily attracted
by the dogs. Our experience as regards adult flies is not in agree-
ment with that of Le Dantec and Boyé; during the dry season,
as we have shown, the adult fly is not rare in Freetown; we
captured over a hundred wild specimens, both male and female;
fertilization and oviposition occurred constantly during the dry
season. There does not seem to be any reason why infection of
man should not occur in this season if the theory that dogs are the
attraction which brings the fly near man, and form the main
reservoir, were correct. That these flies were not, in this case,
attracted by dogs is evident, because there was no dog present in
the bungalow; it was equally clear that they were not attracted by
the latrines, as in no case was a fly ever captured or seen there; nor
were they attracted by the clothes accumulating for the laundry,
which they were never seen to approach. They appeared not to
have come in to lay their eggs, because males as well as females
came in, in about equal numbers and because no female which was
captured was actually ready to lay eggs when it came indoors.
The correct explanation probably is, in accordance with the experi-
mental evidence on the effects of sunlight, that the flies were simply
taking shelter from the heat of the sun; we have pointed out that
on dull days they did not come in. The source from which these
flles came was, we think, the rat holes in the rocks adjacent to the
bungalow, where, as we have pointed out, puparia were found.
Rats captured near the bungalow were frequently infected, and
sometimes heavily.

It might be argued that, although in this case the flies were
not attracted by the presence of dogs, if dogs had been present they
would have induced the flies to oviposit in the house; that might
be so, but if it were the case, how can we explain the absence of
human Myiasis at this time of the year in houses where dogs are
kept at all seasons of the year. The fact that there is a definite

494

wet seasonal incidence of Myiasis in man and also dogs points to
some factor at work which is independent of the presence or absence
of dogs.

One explanation which suggests itself arises out of our discovery
that the wild rat is the chief natural reservoir of the fly; this
explanation is, that the seasonal incidence in man and also in dogs
is dependent on the seasonal habits of the rat. It is commonly
known that in the wet season rats congregate more closely in the
neighbourhood of human habitations ; this movement is due possibly
to the flooding out of their burrows; this theory involves that
Cordylobia moves with the rat, and so is brought into close
association with human habitations. Another explanation 1s that
it is simply the desire to escape from wet which makes the fly lay
its eggs indoors in the wet season.

The flies came in freely, in our experience, during the dry
season, but not with the idea of ovipositing; their normal place for
ovipositing in the dry season is not indoors. We have seen,
- however, that in experiments the fly avoids wet sand and will not
lay her eggs there, she wili rather even lay them on cotton wool and
cloth, and this was clearly done in the case of natural infection
mentioned below. ‘This fact alone might account sufficiently for
the increase in human Myiasis in the rains, and also to a less extent
for the increase in Myiasis of. domestic animals. It is probable,
however, that both factors are at work.

IX. MODE OF INFECTION OF MAN

The earlier erroneous ideas that the fly lays iarvae or eggs in
the skin of man, or that it attaches its eggs to hairs, have already
been referred to. The method of infection in man is by the penetra-
tion of the first instar larvae into the skin; for this the larvae
must effect contact with the skin. It is clear that there are
very many ways in which such necessary contact may be brought
about. It is possible, for example, that where soil or sand,
especially if contaminated, is used in latrines, the female fly may
deposit her eggs in the sand box; if in the act of using the sand
some of it is spilt on the seat of the latrine and first instar larvae

495

are present in the sand, the next person to use the latrine will
almost certainly have the larvae penetrate the skin. Again, if flies
are hard pressed, they will lay their eggs on clean clothes, and
when the larvae hatch out, if the clothes are put on, infection will
occur. An interesting case is that reported to us by Dr. Wright,
of Freetown, a most accurate observer, who has great experience
of this disease; he was called to see a patient who had injured his
shoulder; he wished to put it up at once, and for a bandage was
provided with a window curtain which had been washed and ironed
and had been lying in the house for some time. In a few days,
only under the curtain,. there developed on the patient’s thorax,
back and front, and on the arm, thirty-eight larvae of Cordylobia;
the other curtain which had been lying underneath the first was
examined carefully by us, but no eggs or larvae were present; there
appears here the strongest evidence of the infection from clean cloth.
The probability of the larvae derived from eggs laid on dirty
clothing surviving the washing, exposure to the sun to dry, and
subsequent ironing appears very small. Again, in view of the adult
fly’s dread of the bright sun and the lethal effect which this
produces on the fly, it appears improbable that clothes hung up on
lines to dry in the sun would have eggs deposited on them; if the
clothes were laid on the ground to dry they might pick up larvae
easily from the soil, but would not be so hkely to pick up eggs
which are lying slightly below the surface. In the houses of natives,
the occupants could obtain infection by lying on infected soil, and
also in any of the previously mentioned ways.

xX. SYMPTOMATOLOGY
IN ANIMALS

The presence of one or two larvae produces little obvious distress
even in small animals; when the larvae are numerous, however,
there is very considerable irritation, and the obvious illness of the
animal results chiefly from septic absorption combined with loss
of sleep; the appetite diminishes, and the animal loses weight.
Where larvae are single there is little to note beyond the localized
small tumour; where larvae are close together great swelling and
oedema occur, and the tissues intervening between larvae become

496

sloughing and gangrenous; the rernoval of individual larvae, then,
is difficult, as the whole area of skin surrounding it may come off
with them. The larvae are not always confined to the true skin, but
often penetrate into the deeper tissues; in the case of some of the
guinea-pigs, as we have stated, they exposed the tendons of the
feet. In the abdominal wall the parietal peritoneum may be
involved, giving rise to rigidity of the abdominal muscles, retraction
and tenderness. In certain regions the presence of larvae produces
more serious lesions than in others; the feet and scrotum easily
become gangrenous; the case of one pup which became blind of an
eye was reported to us; the larva had penetrated the skin before
the eyes were open.

IN MAN

The actual penetration of the first instar larva into the skin js
hardly noticeable; in some cases, like the one referred to in a
previous section, an intense cutaneous reaction occurs. Roubaud
observed a similar reaction in his own case, and accounted for it by
saying that the larva had become contaminated from the soil in
which it was. It appears to us more probable that this is a body
reaction in response to the presence of a specific substance produced
by the larva, possibly of a salivary nature, and that it has some
connection with the marked condition of immunity which existed
in one of our human cases. Such a reaction was not observed by
us in the case of another European, recently arrived in this country ;
in this case larvae of the same batch introduced themselves into the
skin without reaction and proceeded to develop. The larva
developing in man 1s felt during the first two days or so, causing
shght itching or pricking at intervals; the symptoms and signs are
easily overlooked. The papule which forms, increases in size and
becomes red; there 1s then a more or less complete cessation of
symptoms for several days possibly, although the furuncular
swelling increases. Then the symptoms recur with greater severity ;
the pain increases and becomes so sharp as to interfere with sleep.
The larva becomes very active at intervals and can be seen clearly
retracting into the cavity and then pushing against the margins of
the aperture in the skin to increase its size. Much serous fluid may
exude at this time; the skin and subcutaneous tissues have meantime

497

become much indurated and the area round the aperture is deeply
coloured ; tenderness on pressure exists; the lesion resembles a boil,
for which it 1s frequently mistaken. Gland enlargement may occur
and general symptoms, malaise and febrile reaction. The develop-
ment was slow in human beings observed; in one case a larva
removed on the fifteenth day in the third instar measured only
gmm. The stage at which larvae are usually brought under notice
by human beings 1s after the third stage has been reached, at which
time the larva, in enlarging the entrance aperture preparatory to
making its exit from the skin, exercises considerable force. The
cavity formation is out of proportion to the size of the larva, and
it appears as if the larva produced a lytic action on the tissues near
its head end; the clear fluid which comes from the cavity at intervals
is sometimes stained with blood and also with faeces of the larva.
On removal of the larva the symptoms disappear, and _ healing
usually occurs readily.

Nagel (1897) observed larvae in his skin in East Africa for a
period of four weeks ; but the record is not complete.

XI. TREATMENT

Various methods of treatment were tried experimentally, such
as the effect of tobacco smoke, insufflation of calomel powder,
French chalk, dropping on tobacco juice, chloroform water, phenol
solution and cresol solution in 5 per cent. strength, application of
vaseline, palm oil and liquid paraffin. Simple expression was
effective in removing the larvae, but often painful; removal by fine
forceps was easy in the later stages if the aperture was large. Of
these various methods the one finally adopted, especially for use
with small larvae which cannot be removed easily with forceps,
was the application of liquid paraffin and subsequent expression.
Blenkinsop (1908) records the effect of a plaster of sugar and soap
in causing larvae to emerge, owing to the blocking of the posterior
spiracles. The natives of Sierra Leone use palm oil, the pericarp
oil of Elaeis guineensis, with the same object; they say, however,
that the Tumbu comes out at night to feed on the oil. Palm oil
was tried, but was discarded owing:to the colour ; in its stead liquid

498

paraffin was used, and acted admirably even with the very small
larvae. A film of paraffin is placed over the opening in the skin,
any scab being first gently removed; at once the posterior end of
the larva begins to come out; the film is then thickened by adding
paraffin drop by drop; the larva in its efforts to reach the surface
of the film makes greater movements out and in; in doing so it
lubricates itself and the walls of the cavity; the superfluous paraffin
is wiped off, and the two thumbs are placed a little distance on
each side of the aperture and pressure inwards and downwards
applied; the larva comes out slowly at first, later with more rapid
movement. The larva should be destroyed. After extraction of
the larva and healing of the wound, a mark remains for a long
time; Fiilleborn (1908) could still after ten years see the marks left

on the skin.

XII. PROPHYLAXIS

Adults should be looked for daily in houses on the ceiling of
rooms and verandah, during the sunny hours of the day; any fly
present should be captured in a collecting net and destroyed. All
latrines should be fly-proof.

Sand and soil used for the latrine may be heated in a kerosene
tin for some time before being placed in the latrine box, in this way
eggs and larvae are destroyed.

In affected districts the weekly examination of domestic animals
should be carefully done, and any larvae found, expressed and
killed ; 1t is most important to destroy the larvae.

Rats should be eradicated as far as is possible from houses and
compounds. Those captured should be destroyed by burning
before the larvae leave them.

In cases of small boils where there is doubt as to the presence of
a larva, the application of a drop of liquid paraffin will cause the
hind end of the larva to move actively; in any case liquid paraffin
will be of great assistance in removing larvae with as little pain as
possible.

_ Clothes lying exposed are a source of danger; it is especially
underclothes and bed linen which are apt to carry infection to man;
a very certain method of prevention here is to have all the clothes

499

ironed after washing and drying, and to store them immediately in
covered receptacies.

It is advisable to have such clothes washed in the compound,
and after 1roning kept in drawers or suitable covered boxes; this
simple precaution will prevent flies laying on them,

XIII. COMPARISON BETWEEN CORDYLOBIA ANTHROPOPHAGA
AND SOME OTHER MYIASIS-PRODUCING FLIES

Myiasis is a wide term embracing parasitism of very varying
degrees, and involving widely different parts of the body. We
shall confine our attention here to those forms of Myiasis caused by
flies which in the first instar have been proved, or appear to be
capable ot penetrating the unbroken skin.

(1) BOOPONUS INTONSUS, Aldrich

Woodworth and Ashcraft (1923) describe a condition of Myiasis
affecting the feet of carabaos and bullocks in the Philippine Islands ;
larvae were reared and adults bred and forwarded to Aldrich.
Aldrich (1923) from three females sent to him created the new
genus Booponus with the species B. iztonsus. He refers to the
close similarity of the adult to Cordylobia. The description of
Woodworth and Ashcraft deals with two larval stages, but the
illustration of the young larva shows that it is not like the first
instar larva of Cordylobia; rather it resembles closely the second
instar of this fly, not only in the appearance of the posterior
spiracles but also in the very large and dark spines irregularly
distributed over the cuticle.

It is surmised, but not proved, that the first instar larva which
arises from eggs attached to hairs is capable of producing Myiasis
by penetrating unbroken skin; if it appears later that this 1s so,
we should expect, on analogy, that a first stage larva having a
buccal spine will be found.

(2) WOHLFAHRTIA VIGIL, Walker

Walker (1920) described two cases of cutaneous Myiasis in
infants due to larvae of this fly; evidence was given of a clinical
character that these larvae appeared to have entered the unbroken

500

skin. Again Walker (1922) gives details of another case and also
descriptions of the first instar; he says ‘the median or labral hook
arises from a slightly divided base, immediately in front of the
pharyngeal sclerites, and is strongly decurved, the pointed apex
projecting slightly from the front part of the oral aperture in the
usual position.’ The illustration shows well this curved hook;
this character differs from the median spine in Cordylobia first
instar larvae; the appearance of this apparatus in WoAlfahrtia vigil
does not suggest that the mode of entering unbroken skin can be
the same as 1n Cordylobia; it is possible that when skin penetra-
tion experiments are carried out, it may be found that this larva,
if it can in fact penetrate healthy skin, does so by digging at once
deeply into the tissue, and not, as in Cordylobia, by raising over
its dorsal surface a thin layer of cuticle.

(3) HYPFODERMA spp.

Laake (1921) gives an account of the anatomy of the mouth
apparatus of 7. bovis and H. lineatum; he refers to the fact that
Riley (1892) first described the real first stage larva of H. lineatum
which he obtained from the egg before hatching; he mentions that
Glaser (1914) and Carpenter, Hewitt, and Reddin (1914) first
observed the first stage larva of H. dbo0vzs outside of the egg.
Laake in his description and drawings shows that the median
mouth spine in these larvae is retained not only during the first
instar but also actually during the second and third instars, during
which the larva is passing through the tissues, and 1s not cast off
until the larva reaches the back of the host. It is interesting to
note that the ventral curvature of the spine is relatively slight
compared with that depicted by Walker for W. vzgzl, and resembles
more the condition present in Cordylobia.

(4) DERMATOBIA CYANIVENTRIS

Surcouf (1913) figures the first instar larva of this fly; the larva
which is able to penetrate the unbroken skin is possessed of a buccal
armature closely resembling that of Cordylobia; the large forwardly
directed cuticular spines on the posterior segments which we drew
attention to in Cordylobia, exist in this larva also.

501
XIV. SUMMARY

1. The morphology and bionomics of Co7zdylobia_ anthro-
pophaga, Griin., have been studied in some detail during the dry
season 1922-23, in Freetown, Sierra Leone. |

2. Certain new facts as regards the habits of the adult, its
method of oviposition and the number of eggs laid by it are
recorded.

3. In the first larval stage also, attention 1s drawn to certain
morphological peculiarities, both in the buccal armature and in the
cutaneous spinulation, which appear to have a direct and intimate
connection with the process of skin penetration.

4. A direct association between Cordylobia and wild rats,
which was suggested by field observation and laboratory expert-
ment, has been proved to exist by the discovery of puparia of this
fly in the burrow of wild rats.

5. Evidence is produced which appears to incriminate the wild
rat as the main reservoir of the infection in nature, and to associate
these rodents with such seasonal incidence of the disease as exists.

6. Numerous experiments were carried out both in man and
animals, which add considerably to the knowledge of the mechanism
of infection, pathogenicity and prophylaxis.

7. An immunity has been proved experimentally to develop
against attacks of the larvae, not only in man but in animals.

8. The development of such an immunity 31n the case of cattle
in similar forms of Myiasis 1s considered a possibility and worthy
of investigation.

g. A comparison is made between Cordylobia and some other
flies which cause cutaneous Mylasis.

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SaMBoNn, L. W. (1915). Rep. Adv. Comm. Trop. Dis. Res. Fund for 1914, p. 139.

Sir, F. (1908). “Fl. R.4.M.C., Vol. X, p. 14.

Surcour, J. (1913). C.R. Acad. Sct. d. Paris, Vol. CLVI, p. 1406.

Watxer, E. M. (1920). ‘Fl. Parasitol., Vol. VIT, p. 1.

(1922). ‘fl. Parasitol., Vol. IX, p. 1.

WoopworrtH, H. E., and Asucrart, J. B. (1923). Philipp. Fl. Sct., Vol. XXII, p. 143.
ZePeDA, P. (1913). Rév. de Méd. G d’Hyg. Trop., Vol. X, p. 93.

504

EXPLANATION QT SPicA PE ax

Cordylobia anthropophaga, Adult 3 and Q.

Annals Trop. Med. & Parasitol., Vol. XVII

Fea EF LIAS 6

del. M.G.T.

4SV¥ RATAN * ) MOITAVA. CPAs

5. | 5
abs to 2orse Vo dgargmodT “ oA OVAL Bovws: >

+ BITRE

506 — :

EXPREANATION OP (PICA Tieacy

Cordylobia anthropophaga. Photograph of series of adults and

puparia.

Annals Trop. Med. & Parasitol., Vol. XVII PIAL YX Ll

Photo. by M. Brown

C. Tinling & Co., Lid., Imp.

¢ Lea AR,
it THE
weriknoliY OF Mbinais

508

EXPLANATION: OF SPLAT Hae val

Photograph of naturally infected rats (shaved with Barium

depilatory powder).

mals Trop. Ved. Dj Parasitol., Vol. XVII PLATE OL

| boto. by M. Brown 7
C, Tinling & Co., Lid., 1mp.

510

EXPLANATION OF -ERAVTiIoe sin

Cordylobia anthropophaga. Photograph of second instar larva in

the tissues of a guinea-pig. x 8.

Annals Trop. Med. & Parasitol., Vol. XVII eye ae. en MUA

Photo. by M. Brown

C. Tinling & Co., Ltd., Imp

rat TIBSARY
Of iHE
IMIWERSITY OF MM hawuls

511

THE TRANSMISSION OF
T. CONGOLENSE BY GLOSSINA
PALPALIS

Pion Ooo lah BO) BoE. ML) 4 Se.D. Cantah:

BACTERIOLOGIST, UGANDA PROTECTORATE

(Received for publication 10 October, 1923)

CONTENTS

1. INTRODUCTION na 511
2. Previous Work ON THE ‘TRANSMISSION OF THE PROBoscIs-AND-GutT GRouP OF

TRYPANOSOMES BY G. palpalis 514
3. History oF THE TRYPANOSOME-STRAIN EMPLOYED IN THE TRANSMISSION ExPERI-

MENTS PERFORMED AT ENTEBBE DURING 1922 AND 1923 = ne ae ci
4. ANIMAL REACTIONS OF THE STRAINS OF T. congolense REFERRED TO IN THIS PAPER... 519
5. ‘TECHNIQUE 520
6. ‘THe TRANSMISSION EXPERIMENTS ... - + 34 ow ids a5 521
7. Discussion OF THE EXPERIMENTS aa a <a ar a a a 527
8. SUMMARY 529
g. REFERENCES nee a ee aon de sos nes sa nee Hi 530

I. INTRODUCTION

Representatives of the group of trypanosomes to which
I. congolense and IT. nanum belong are widespread throughout
Africa. [hey are common alike in game and in stock. To the
game they are, apparently, harmless, but as parasites of domestic
animals they constitute a factor of considerable economic importance.

The trypanosomes of this group are well adapted to rapid
propagation in nature. For indirect or cyclical transmission they
depend, so far as is known, entirely on the Glossznae; and, though
less common than 7. vevax and 7. uniforme in wild game-tsetses,
they are considerably more prevalent in the fly than the polymorphic
trypanosomes of the 7. bvucez group. In all species of Glossinae
in which cyclical development of the 7. congolense group takes
place, the flagellates multiply first in the gut of the fly, and later
on take up their ‘ anterior station’ in the labrum and hypopharynx
of the proboscis.

In addition to cyclical transmission, they are readily conveyed

512

from mammal to mammal by the direct method—that is, without
any alternation of generations. It is probable that most, if not all,
the mammalian trypanosomes can, on occasion, avail themselves of
direct transmission, but it is doubtful whether any other group of
the Glossina-carried organisms relies to the same extent on this
method in nature.

The so-called species contained within this group have been
arbitrarily separated on account of differences in their behaviour
in certain mammals. The xanum-form cannot infect dogs and
monkeys and the small laboratory animals, while the congolense-
form can; otherwise they are indistinguishable. Both these two
forms of the trypanosome have been recovered from game and from
wild fly, so that this idiosyncrasy may, to a slight extent, influence
the distribution of the parasites in certain regions. But to allow to
physiological variations of this kind the importance attaching to a
specific character, 1s, as Yorke and Blacklock (1914) have pointed
out, unjustifiable; and, as a matter of fact, strains whose virulence
is intermediate between these two extremes do occur in nature. The
group should, therefore, be regarded as a collection of strains whose
morphology and behaviour in the insect intermediary are uniform
and constant, but which vary in their relation to certain mammalian
hosts. It has been shown elsewhere by laboratory experiments, that
the effect of long-continued direct transmission on a strain of
IT. brucez may be to enhance its virulence in mammals, and, at the
same time, to eliminate the power of developing cyclically in
Glossinae. The same principles probably apply to the group we
are considering. There 1s, indeed, field-evidence to show that some
of the more virulent strains of 7. congolense rely largely, if not
solely, on direct transmission, for their passage from mammal to
mammal. Whether such strains retain the power of cyclical develop-
ment in Glossinae has, however, not yet been determined.

The gregarious habits of domestic stock afford excellent
opportunities for the spread of their trypanosomes by direct
transmission, by Stomoxys and the other species of biting flies that,
in Central Africa, swarm around the unfortunate animals. Among
the game this method functions probably much less commonly,
although with certain species such as elephant, buffalo and eland
the requisite conditions will doubtless often occur.

513

However this may be, there is an essential difference between
game and stock in their relation to these parasites. For while the
game is not, so far as is known, in any way inconvenienced even
by double infections of the trypanosomes indigenous to its habitat—
whether directly or cyclically transmitted—cattle, as a rule, cannot
survive even brief contact with the game-tsetses, and are particularly
sensitive to strains of 7. congolense in non-tsetse areas. In
localities where game-tsetses are present and game plentiful, a
certain percentage of the wild fly will always be found to carry the
developmental forms of the proboscis-and-gut group of trypano-
somes. In G. pfalpalis areas, however, matters are different. The
commonest flagellate found in this fly in nature is probably 7. gvayz.
Of the mammalian trypanosomes, the proboscis-only group is
most commonly found, represented by 7. vivax and T. uniforme ;
the gut-and-gland group, usually in the form of 7. gambiense,
is also of common occurrence; but the proboscis-and-gut group is
distinguished by the comparative rarity with which it develops in
wild G. palpalis.

So far as I can determine, three instances are recorded of the
occurrence of developmental stages of the proboscis-and-gut group
in wild G. palpalis. These are as follows :—Warrington Yorke and
Blacklock (1915), investigating an epidemic of cattle trypanosomiasis
in Sierra Leone, found nineteen out of one hundred and forty-seven
cattle infected with trypanosomes; 7. congolense was found in
sixteen, 7. vivax in seven, and 7. gambiense in one. In the course
of the dissection of four hundred wild G. palpalis caught in the same
neighbourhood, proboscis-only infections were found in fifteen flies,
gut-only in two, and gut-and-proboscis in four.

Macfie (1915) dissected seventy-five wild G. falpalis from near
Accra, and found eleven infected with flagellates—three gut-and-
gland, three proboscis-only, and one proboscis-and-gut. / In the
review of these experiments, there follows the seemingly contradictory
comment that ‘none of these infections resembled stages in the
development of 7. congolense.’ Possibly the flagellates in the
proboscis of the single gut-and-proboscis fly were not fixed, and, in
consequence, were not regarded as true proboscis-forms.

The third instance occurred in my own work (1916) in the
Northern Province of Uganda, where, in two areas, proboscis- y,

514

and-gut infections of wild G. palpalis were found, amounting in
one case to I°4 per cent. and in the other to 5°2 per cent. of the flies
dissected. Allowing for the possible occurrence in some of these
flies of double infections, the last figure is sufficiently high to prove
conclusively that, in this area, the congolense-nanum type of
trypanosome 1s carried cyclically by G. falpalis.
~* In the face of these examples it is the more remarkable that no
instance has hitherto been recorded of the occurrence of this group
of trypanosomes in the wild G. palpalis of the Victoria Nyanza. At
many points along the shores of this great lake, cattle came in contact
with this tsetse; and there are long stretches of shore in Busoga
where game and G. pallidipes occur immediately behind the
G. palpalis shelter, and where the latter fly has plenty of opportunity
for picking up parasites of the 7. congolense group. T. vivax,
IT. untforme, and a member of the gut-and-gland group are all found
in these lakeshore G. falfalis. During the fourteen odd years that
have elapsed since the depopulation of the Sesse Islands, the
Situtunga have multiplied enormously, and the tsetses on these
Islands have come to rely on the antelope to a great extent for food.
The three trypanosome species just mentioned, are common in these
Situtunga, but on no occasion has the 7. congolense type been found
in their blood. Apparently G. palpailzs, in its natural environment,
is not, as a rule, suited to the cyclical transmission of these
trypanosomes.
A possible explanation of the absence of the proboscis-and-gut
group from the fly on the Victoria Nyanza is, that the strains with
which the fly come in contact are directly transmitted strains, which
have lost the power of cyclical development in Glossivae. But this
explanation alone is inadequate to account for the absence of these
parasites from this huge fly area.

Il. PREVIOUS WORK ON THE TRANSMISSION OF THE
PROBOSCIS-AND-GUT GROUP OF TRYPANOSOMES BY
G. PALPALIS

The Royal Society’s Commission in Uganda (1910) experimented
with wild and with laboratory-bred flies. Two successful
transmissions were reported. In one experiment, with wild
G. palpalis, the flies became infective to a clean animal twenty-one

515

days after the first infecting feed; in the other, in which laboratory-
bred flies were employed, the flies were infective on the fourteenth
day of the experiment. The authors remark that these last experi-
ments were open to fallacy, as an epidemic of trypanosomiasis, due
to a member of the ‘ proboscis-and-gut’ group, was occurring at the
time in the neighbourhood of the laboratory. I think it probable
that the same explanation applies to the wild fly transmission also.
No positive flies were found on dissection of the flies in these
experiments, and at the time, it was not known that the develop-
mental cycle of 7. congolense included an invasion of the proboscis
of the fly. Subsequent experience has shown it to be improbable
that the trypanosomes of this group can complete their cycle in
G. palpalis in such a short time as twenty-one days.

Of the attempts by the same Commission to transmit 7. xanusi
by G. palpalis, we read that the only experiment attempted was
‘unsatisfactory, as trypanosomes appeared in the first healthy goat
a few days after the fly had fed on infected animal,’ 7.e., it was a
natural infection due to some agency outside the experiment.

In view of the ease with which these ‘ proboscis-and-gut’
trypanosomes are propagated in nature by agents other than tsetse-
flies, 1t 1s, In my opinion, unsafe to carry out experiments at a spot
where the disease is already existent, especially if ruminants are
employed to demonstrate the transmission.

In 1911, Fraser and Duke carried out seven transmission
experiments with laboratory bred flies. One experiment was
successful, trypanosomes appearing in the blood of the clean monkey
on the eighty-fifth day after the first infecting feed of the flies. On
the ninety-sixth day of the experiment a fly died which showed a
heavy proboscis-infection. This fly, after infecting the clean
monkey, had had access to two other clean monkeys, upon one of
which it certainly fed several times. Neither of these last two
animals became infected. At the time, it was supposed that the
insect must have lost its infectivity, but I now believe that the
explanation lay in the different resistance of the host-animals.

In the dissections of these experiments, twenty positive flies were
found in a total of four hundred and twenty-seven dissected.
Among these were five flies with flagellates established in the
proboscis, One of these infected the monkey in the positive

516

experiment. Two occurred in another experiment, and_ fed
repeatedly on a clean monkey without causing infection; they never
had access to a ruminant animal. The remaining two flies with
proboscis-infections occurred in a third experiment, and died on the
fiftv-ninth and seventy-fourth days after the original infecting
feed; they both fed repeatedly on a clean monkey without ever
infecting it.

The contents of the proboscis of all these five positive flies were
injected into rats, without infection resulting. In these experiments
no infection of the proboscis was met with in flies dissected before
the fiftieth day after the first infecting feed. The salivary glands
of these positive flies were all negative to flagellates. In all these
experiments the clean animal employed was a monkey. No
trypanosome disease existed in the vicinity of the laboratory at the
time.

In 1911-12 experiments were carried out on the transmission of
L. nanum by laboratory-bred G. palpalis in Uganda. In the first
set of experiments a goat was used for the infecting feeds;
one hundred and seventy-three flies were dissected during these
experiments; no infected flies were obtained and no transmission
occurred.

In the next series of experiments an infected sheep was employed,
on which the flies fed much more readily, and the parasite was
transmitted to a clean calf. Three hundred and twenty-two flies
were dissected, of which twelve were infected, five showing flagellates
established in the proboscis. In only one of the proboscis-infections
were flagellates seen in the hypopharynx; the labrum of this fly
contained great clusters, while a few individuals were present in the
hypopharynx. This fly died on the twenty-fifth day after the first
infecting feed—the earliest recorded date for the infection of the
proboscis of G. palpalis by a member of this group of trypanosomes.

In 1git-12 another series of transmission experiments were
carried out with 7. congolense and laboratory-bred G. fpalpalzs.
The animal used for infecting the flies was a young bushbuck which
had been born at the laboratory and, when a few months old, had
been infected by syringe inoculation with the Mpumu Laboratory
strain. In the course of these experiments, seven hundred and
forty-six flies were dissected, of which six hundred and thirteen lived

517

until the thirtieth day after their first feed on infected blood.
Fifteen flies were found to contain flagellates; four had proboscis-
infections; and one a heavy infection of the sucking-stomach, with
no flagellates in the proboscis. The box which was apparently
responsible for the successful transmission, contained this fly with
the infected sucking-stomach and none with infected proboscis. The
four flies with infected proboscides died respectively on the
seventy-sixth, one hundred and fourth, and one hundred and forty-
first days after the first infecting feed. A ninetieth day fly showed
flagellates in the hind-gut only.

It must be noted that the strain of 7. congolense kept at the
Mpumu Laboratory was derived from cattle, from localities where
tsetse-flies are absent or scarce. It is, therefore, probable that it
was a directly transmitted strain, before it commenced its career at
the laboratory. Too much stress, therefore, cannot be laid upon its
behaviour when exposed to tsetses.

The strain of 7. xanum, on the other hand, came from cattle
in the G. pallidipes country in Toro Kingdom, and Sheep
Experiment 59, by which the flies were infected, was the second
passage animal from the original ox.

As already stated, the strain of 7. congolense used in the
experiments now to be set forth is known to be a cyclically carried
wild fly strain.

III. HISTORY OF THE TRYPANOSOME STRAIN EMPLOYED
IN THE TRANSMISSION EXPERIMENTS PERFORMED AT
ENTEBBE DURING 1922 and 1923

This strain was brought back from Mwanza, in August, 1922, in
Bitch Experiment 649.

On August Ist, 1922, Experiment 649 was fed upon by a batch
of wild G. swynnertont, of which two, on subsequent dissection, were
found to have proboscis-and-gut infections, the salivary glands
being negative. On August 8th, 1922, a single trypanosome of the
T. congolense type was seen in the animal’s blood. On August oth,
10th and 11th, the blood was negative to fresh film examination ;
on the 12th a few trypanosomes were seen, and thereafter the
animal was negative to daily examination of stained thick films

518

until September 13th. From this date trypanosomes were found at
frequent intervals in the course of the daily routine examinations,
until February 2nd, 1923. Then followed a series of negative
examinations until February 28th, when a few parasites were seen in
a stained thick film. Trypanosomes were next seen, on a single
occasion only, on August 20th, 1923. For a month or so after
arriving at the Entebbe Laboratory, the animal was languid and
seemed to be losing condition. In October she showed signs of
being pregnant ; the mammae enlarged and the animal became fatter,
but in a week or two these symptoms disappeared. The general
condition was now excellent. She became pregnant about the
middle of June, and on August 24th, 1923, at 12 o’clock, two
healthy puppies were born, followed, an hour and a half later, by
a third which died an hour after birth.

The subsequent upkeep of the strain from this bitch is shown

in Table I.
Tasze I.
Bitch 649, infected by wild G. scvynnertoni
V V
Calf 644 Monkey 554
(infected by laboratory-bred (infected by laboratory-bred
G. palpalis, Experiment 536) G. palpalis, Experiment 536)

| |

V
Monkeys 672 and 673 Calf 690 Monkey 671 Monkey 68:
(inoculated by syringe) _ (infected by laboratory-bred _ (infected by laboratory-bred _(infected by laborat)
G. palpalis, Experiment 686) G. palpalis, Experiment 660) G. palpalis)
seul Vv
Guinea pigs 723 and 724 Monkey 722

(inoculated by syringe; never (inoculated by syringe)
showed trypanosomes)

LESS

519

IV. ANIMAL REACTIONS OF THE STRAIN OF T. CONGOLENSE
REFERRED TO IN THIS PAPER

The 7. congolense isolated by Bruce’s Commission and employed
in their original transmission experiments at Mpumu, lost some of
its virulence during two years of maintenance at that laboratory.
The average duration of the disease in ten completed monkey
experiments carried out by Bruce was sixty-three days; one animal
died after one hundred and eighty-one days, the average for the
other nine being forty-eight days. Another monkey was alive after
two hundred and sixteen days. In eight completed dog experi-
ments the average duration was forty-three days. Of three monkeys
inoculated with the same strain eighteen months later, one died after
thirty-five days, one after one hundred and fifty days, and the other
was still alive after two hundred and three days.

The animal reactions of the strain from the Mwanza Fly Belt are
as follows :—

Taste II.
Incubation period, Duration of disease,
in days in days
Monkeys— ;
554 ++ ..«| 78 days or more... zeal 127 ats least,
9 eae) 47 Gaye OF more. ... ...| Still alive after 145 days.
680). nae ? 47th day after first appearance of trypanosomes
of Fly Experiment 686.
672):00 or le ee ie oa ...| Still alive after 188 days.
OFF cs ead are bik a ...| 64 (trypanosomes swarming in blood).
ip ere 1 py ihe they ee ys age ...| 43 (trypanosomes swarming in blood).
Calves—
640 ... Sasha lOy sors bet Eee ...| Alive and apparently well after 7 months.
Cy 7 oe ry ae oe Jess ...| Alive and apparently well after 9 months.
690 ... eait Tages ve nf ...| Alive and apparently well after 107 days.

§20

V. TECHNIQUE OBSERVED IN THE CONDUCT OF THESE
TRANSMISSION EXPERIMENTS

The transmission experiments were carried out according to the
methods pursued for many years in the Uganda Laboratory. The
newly hatched flies are placed in wire-sided boxes and fed and’
starved on alternate days, dead flies being removed for dissection
each morning. During the act of feeding, a wet rag covers the top
side of the box, while the other side is closely applied to the animal’s
skin. Feeding continues until the flies have had all the blood they
want. At the termination of the experiment the survivors are killed
with chloroform vapour and dissected. Experience has proved that
the rough and ready method of holding the box in the smoke of a
fire, though effective in killing the flies, also kills the flagellates
they contain, and makes the identification of light infections more
difficult. Throughout the experiments the boxes are kept on stones

resting in dishes of clean water.

Whenever a fly with a proboscis-infection was found, the animals
upon which the insect had fed were examined daily by means of a
stained thick blood-film. Otherwise, all experimental animals were
examined daily by careful inspection of fresh unstained blood-films.

VI,

The actual experiments will now be set forth.

571

THE TRANSMISSION EXPERIMENTS

In the ‘ positive-

flies’ column the contents of the brackets refer to the number of

flies with infected proboscides.

In the ‘remarks’ column will be

found the result of the experiment, positive or negative, according

as the clean animals develop trypanosomes or not.

(A) Experiments in which some of the flies developed proboscis-infections.

(1) Experiments in which cyclical transmission of 7. congolense

from sick to healthy animal occurred.

Taste III.

EXPERIMENT 536.

Day of
Date Experiment
1922
Sept. 25-27 | 1-3
» 28 4
» 29-Nov. 21 5-58
Noy. 22 59
, 23-Dec.1 | 60-68
Dec. 2 69
oe 34+ Fou
a) 1s

Procedure

Flies fed on bitch 649
Starved

Fed on clean monkey 554
Starved

Fed on clean calf 644 and
on 554 onalternate days

Starved
Fed on clean monkey 611

Remaining 28 flies dis-
sected

Tee. |
Positive flies found |

2
(1 proboscis +)

3
(1 proboscis +)

Remarks

T. congolense not seen in stained
thick films of bitch’s blood.

T. congolense first seen in 554’s
blood on 13 Feb., 1923.

T. congolense first seen in calf’s
blood on Dec. t.

/
Monkey 611 never showed
| trypanosomes.

Flies dying before 16th day were ignored.
Total number of flies dissected during the experiment = 68.
Number alive on 25th day of the experiment = 64.

Remarks. November 27th was the last date on which the flies
had access to 554, the incubation period in the monkey was thus
very long, amounting to at least seventy-eight days. In contrast
to this the incubation period in the calf was only nine days.

522

Tasre IV.

EXPERIMENT 660.

Date
1923
Feb. 16
» 17
»» 18-19
Rage Se:
Pm A a 5
3) 24

»5 25-April 8

April 9-10
“Pek

Experiment

Day of

Flies dying before the 22nd day of the experiment were ignored.

Procedure Positive flies found

Flies fed on monkey 554

Starved ° es
Fed on monkey 554

Starved

Fed on monkey 554

Starved
Fed on clean monkey 671 I
on alternate days (proboscis +-+-++)
sist day
Starved nos “ as
Remaining 19 flies dis- a
—sected

Total number of flies dissected = 58.
Number alive on 25th day of experiment = 55.

Remarks.

May.

Note the long incubation period of 7. congolense in
monkey 671, i.e., from, say, the first week in April until 24th of

Remarks

T. congolense present in 554’s
blood.

T. congolense present in 554”

blood.

T. congolense present in 554’s
blood.

Monkey 671 first showed
T. congolense on 24 May, 1923

523

TABLE V.

EXPERIMENT 686.

Day of
Date Experiment Procedure Positive flies found
1923
ar. II-13 I-3 Flies fed on monkey 554.
9 14-15 4-5 Starved
y 16-May 13 | 6-64. Fed on clean monkey I
: | (proboscis +-)
54th day
May 14 65 Starved ee
HS 66-68 Fed on clean calf 690 |
» 18-19 | 69-70 Starved ,.. a ha I
) (proboscis +)
69th day
7” 20 | 71 Fed on 689 Tes
|
% 21 | 72 Starved
pe 22 73 Fed on 689
23-24 74-75 Starved "%. nate or aed
, 25 76 Remaining 14 flies dis-
| sected

monkey 689.

Flies dying before 12th day were ignored.
Total number of flies dissected = 49.
Number alive at 25th day = 46.

Remarks

—————__—_.

}
!

{

T. congolense present in 554’s
blood.

Monkey 689 first showed
T. congolense in blood on
23 May, 1923.

Calf 6go first showed T, con-
golense in blood on 7 June,

1923.

Remarks. It 1s impossible to estimate the incubation period in

twenty-five days after the infecting feed.

In the calf, trypanosomes appeared twenty-three to

524

(2) Experiments in which no transmission occurred.

Taste VI.

EXPERIMENTS 597-8.

Day of
Date Experiment Procedure Positive Flies found Remarks
1922 . ;
Nov. 18 I Flies fed on dog 649s... <i I. congolense not seen in dog’s —
blood.
3 29 2 Starved
9) 20-21 3-4 Fed on dog bes ry: ene T. congolense not seen in dog’s
blood.
Ne 23 5 Starved
sp 237 DeCu2S 6-38 Fed alternate days on ie Monkey 602 never showed try-
clean monkey 602 panosomes: examined by
thick stained films till 15 June,
Dec. 26 39 | Starved ... ose oe Ye 1923.
a os 40 | Fed on clean dog X__... cc Dog X never showed trypano-
somes.
ee es) 41 Starved
sO 42, | Fed on dog X
4 82 43 Starved 2 This fly had fed on dog X.
(1 proboscis ++)
1923
9» 31-Jan. 16 44-60 Fed on alternate days on
monkey 602 and dog X
jan, “17 61 Remaining 32 flies dis-
sected

T'lies dying before the 11th day of the experiment were ignored.
Total number of flies dissected = 113.
Number alive at 25th day of experiment = 105. ©

Remarks. Neither of the clean animals of this. experiment
became infected. The infection of the proboscis in the forty-three
day fly was not heavy, and may have been established after the fly
had been removed from contact with monkey 602.

Possibly, in the earlier stages of the invasion of the proboscis,
the fly is only capable of infecting very susceptible mammals, on
account of the small number of trypanosomes which it inoculates.

ay ae

ett

24-April 21

April 22

Remarks.
repeatedly on monkey 668 without causing infection.

525

Tasre VII,

ExveRIMENT 658,

Day of
Experiment Procedure
| See
I-2 Fed on monkey 554
3 Starved
4 Fed on monkey 554
5 Starved
6 Fed on monkey 554
7 Starved
8 Fed on monkey 554
9 Starved
10-66 Fed on clean monkey 668
67 Starved
68-70 Dog Y
71 Starved
72-76 Fed on monkey 668 and
starved alternate days
77-78 Fed on clean calf
79-81 Starved
82 Remaining 12 flies dis-
sected

f
Positive fies found |

‘proboscis ++)
66th day

Remarks

_T. congolense present in
blood.

_'T. congolense present in

blood.

| ;
T. congolense present in

blood.

}
| T. congolense present in 5654's

blood.

Monkey never showed trypano-
somes: examined daily,
stained thick films till 23 July,
1923.

Dog never showed trypano-
somes.

Calf never showed trypano-
somes.

Flies dying before the 17th day were ignored.

Total number of flies dissected = 41.

Number alive on 25th day = 47.

The omy _ positive

fly of

this

experiment fed
Unfortunately

this fly died before feeding on the dog or the calf.

526

(B) Experiments in which no flies with infected proboscides were found.

(In none of these experiments did transmission occur.)

Table VIII gives a summary of these experiments.

mes aa aha ‘ok
§20 19-21.9.22
600-1 22-24.11,02
643 g, Io and 12.2.23
662 18-21:2.23
665 DH ore Bed
664 19-21 and 23.2.23
667 23-25.2.23
674 1-3.3-23
678 3-6.3.23
084 8-10.3.23
693 16-1 7.3.28
700 19-22.3.23

The results of dissection of the positive flies of all the above

Infecting
animals

.| Bitch 649
.| Bitch 649

Calf 644
Calf 644

.| Calf 644
Monkey 544...
.| Calf 644

Calf 644

...| Calf 644
.| Monkey 554...
.| Monkey 673...
..| Monkey 554...

‘Tasie VIII.

NuMBER oF FLiEs

Alive
25th day

experiments are shown in Table LX.

Dissected
during
Experiment

63
123
66

Containing

flagellates
in gut

Day of

Duration of | Experimen

Experiment
in days

on which
dissectior
began

18th
13th
16th4
13th
24th
18th
rith
15th
rgth
16th
roth
14th |

527

TaBLe IX.

Experiment

Number
of fly

Day of
dissection
reckoned
from first
infection

feed

20th day
21st day
26th day
34th day
43rd day
43rd day
sist day
52nd day
53rd day
54th day
§5th day
§5th day
57th day
58th day
62nd day
66th day
69th day
72nd day
72nd day
72nd day

Distribution of flagellates

Proboscis

(@)

Remarks

Sucking stomach +--++-+

VIF.

DISCUSSION OF THE EXPERIMENTS

It will be seen from Table IX that the earliest date at which
flagellates were found in the proboscis was in a forty-three day fly.
It is, however, possible that the invasion of the proboscis in some of
the older flies took place éarlier than this.

On the other hand,

528

several infected flies lived considerably longer than forty-three days
without the flagellates reaching the ‘ anterior station.’

‘The experiments of Group A show that flies with heavily infected
proboscides may feed upon a clean animai wiitiout causing infection.
It is interesting to note that, when this happened, in each case the
clean animal was either a monkey or a dog: whenever a calf was
bitten by a fly with an infected proboscis, the animal developed
trypanosomes.

It was hoped, by exposing, aiternateiy, different species of clean
animals to the infected flies of these experiments, to throw light upon
the biological significance of the differences in virulence and of
host proclivities shown by the trypanosomes of this group. But,
unfortunately, on several occasions the infected fly died before the
box was transferred to a second, or a third clean animal. It was
thus not possible to ascertain whether a fly may be infective to a
ruminant and yet be unable cyclically to infect a monkey or a dog.

The strain of 7. congolense used in these experiments was of
comparatively low virulence. On several occasions the incubation
period in monkeys, before trypanosomes appeared in the peripheral
blood, was very long. In the case of monkeys Nos. 554 and 671,
both of which were infected by flies, the incubation periods were
seventy-eight and forty-seven days, respectively; with monkey
Experiment 722, infected by the syringe, thirty-seven days elapsed
before trypanosomes were detected. The incubation periods in the
calves infected by flies were nine and twenty-three to twenty-five
days. Similarly, in my experiments at Mpumu, the incubation
period in the only monkey infected by cyclical transmission was
thirty-three days, while in another monkey, infected by means of the
syringe, the incubation period was forty-seven days.

In contrast to this, the average incubation period in thirteen
monkeys inoculated by Bruce (1910) in Uganda was 12°3 days,
maximum twenty-one days. Bruce was employing a virulent
strain—probably directly transmitted—derived from a_ cattle
epidemic. This strain was maintained for upwards of two years by
syringe inoculation from monkey to monkey before being subjected
to the transmission-experiments just referred to.

Both the xanum and the congolense-forms of this group—
distinguished from one another by their different behaviour in

529

monkeys and dogs—have been recovered from wild game tsetses,
the xanum-form being the most common. The congolense-form is
acknowledged to be the more virulent, and is almost always present
in cattle epidemics due to this group of trypanosomes. The evidence
supplied by the experiments set forth in Section 6, taken in
conjunction with the observations already recorded on this group of
trypanosomes, suggests that cyclical transmission of 7. congolense
tends to the acquisition by the trypanosome of a low degree of
virulence, which may be associated with inability to infect such
animals as monkeys and dogs in nature. Directly transmitted
natural strains, on the other hand, usually possess greater virulence,
and readily infect these two animals.

SUMMARY

1. Three out of seventeen cyclical transmission experiments,
performed with laboratory-bred G. palpalis and a wild fly strain of
I. congolense, were successful. In the course of these seventeen
experiments, one thousand and fifteen flies were dissected, of which
eight hundred and ninety-three lived until the twenty-fifth day after
their first feed on an infected animal; eight of these flies had
flagellates established in the proboscis.

2. In all the successful transmission experiments, one or more
flies with proboscis-and-gut infections had fed upon the clean
animals which acquired infection. There is every reason to believe
that these flies were responsible for the transmission.

3. Flies with equally intense proboscis-and-gut infections were
found in two experiments in which no transmission occurred.
Several of these flies had fed repeatedly on clean monkeys without
infecting the animals; in no case, however, did a fly with a proboscis-
and-gut infection feed upon a clean calf without infecting it. It
would appear, therefore, that monkeys are less susceptible to this
strain of 7. congolense, carried by G. palpalis, than are calves.

4. In several cases the incubation period in the monkey was
very prolonged.

5. The strain of 7. congolense used in the experiments is less
virulent than the strain used by Bruce at Mpumu, in Uganda.

530

The Mpumu strain was almost certainly directly transmitted before
its arrival at the laboratory, while the strain here described was
carried cyclically by wild tsetse. It 1s possible that there is a
definite relation between the virulence of a strain and its method of
transmission.

6. The apparent fact that the wild G. palpalis of the Uganda
shores of Victoria Nyanza do not carry trypanosomes of the
proboscis-and-gut group is to be explained, in part at any rate, by
the partiality of the fly for animals which are not susceptible to this
group of trypanosomes. It would appear, however, from the
experiments above described, that this G. falfalis is less fitted to
act as a true intermediate host of the Z. congolense group of
trypanosomes than of 7. vivax, T. untforme, and T. bruce.

My thanks are due to Dr. Mary Martin, Assistant Bacteriologist,
Uganda Protectorate, for valuable help in the conduct of these
experiments,

REFERENCES

Bruce, D., and orners (1g10a). The Development of Trypanosomes in Tsetse Flies. Proc.
Roy. Soc., Series B., Vol. LXXXII, pp. 368-388.

— (1g10b). ‘Trypanosome Diseases of Domestic Animals in Uganda. Trypano-
some pecorum sp. nov. Proc. Roy. Soc., Series B., Vol. LXXXII.

Duke, H. L. (1912a). The Transmission of Trypanosoma nanum (Laveran). Proc. Roy. Soc.,
Series B, Vol. LXXXYV, pp. 4-9.

—-—— (1912b). Some observations on Trypanosoma pecorum (Bruce) and T. uniforme (Bruce).
Proc. Roy. Soc., Series B., Vol. LXXXV. pp. 554-561.

——— (1916). Trypanosomiasis in Northern Uganda. ‘ournal of Hygiene, Vol. XV., pp.
372-387. 7

—-—— (1923). Further Inquiries into the Zoological Status of the Polymorphic Mammalian
Trypanosomes of Africa, and the Means by which they are spread in Nature. Parasit-
ology, Vol. XV., pp. 258-295.

Fraser, A. D., and Duke, H. L. (1912). The Development of Trypanosomes in Glossina
palpalis. Reports of Sleeping Sickness Commission of the Royal Society, No. 12, pp. 47-52.

Macrir. J. W. S. (1917). The Results of Dissections of Tsetse Flies at Accra. Report of the
Accra Laboratory, pp. 49-54. (Summarized in Trop. Dis. Bull , Vol. IX, p. 340, 1917).

Yorke, WARRINGTON, and Bracxtock, B. (1914-15). The Differentiation of the more
important Mammalian Trypanosomes. Ann. Trop. Med. & Parasitol., Vol. VIII,
pp. I-11.

———+ ——— (1915). Notes on Certain Animal Parasites of Domestic Stock in Sierra Leone.
Ann. Trop. Med. & Parasitol., Vol. IX, pp. 413-420.

531

ON A NEW SPECIES OF PHLEBOTOMUS
FROM JAPAN

RONDE Wo-beAD, “Rio.

(Received for publication 15 October, 1923)

Phlebotomus squamirostris, n. sp.

Superior claspers of the male with two pairs of stout spathuliform
spines, the proximal pair arising from a well-developed tubercle.
Terminal segment of the palpi slightly longer than the fourth.
Rostrum (labium) densely scaly.

Male. Abdominal hairs recumbent and _ uniformly pale
ochraceous as on other parts of the body. Wings faintly iridescent ;
costal hairs scarcely darker than the rest. Rostrum ‘(labium)
(fig. 14) densely clothed with long, forwardly directed, non-
deciduous scales. Antennae with relatively long segments, and
short, unilateral, geniculated spines; third segment (fig. 1 @)
projecting beyond the tip of the proboscis to a distance of o'1 mm.
or more, the geniculated spine placed near the distal fourth. The
spine on the other segments is placed proximally a little in advance
of the articulation (fig. 1 6). Palpi rather robust; the third segment
distinctly incrassate, the fifth about one-third longer than the fourth ;
formula I (2, 4), 3, 5. Wings (fig. 1¢) lanceolate; the anterior
branch of the second long vein about equal in length to the distance
between the forks. Armature (fig. 1 2); superior claspers with two
pairs of stout spathuliform spines, arranged in two pairs; the first
attached to a well-marked tubercle slightly beyond the middle of
the segment; the second pair terminal, each arising from a tubercle.
Inferior claspers relatively short, and either equal to, or very
shightly longer than the proximal segment of the superior claspers.

532

Length: 2°7 mm.; length of wing: 1°6 mm.; antenna: 2°3 mm.

Female. Arrangement of abdominal hairs and colour as in the
male; but the body 1s shghtly more robust.

JAPAN :—Agori: July 19th, 1916, 1 @ (Dr. Shinichiro Yamada) ;
Matzuyama: June 25th, 1916, 1d, 1 9 (Dr. S. Komatsu),

/ mm. (s

en — EL

Fic. 1. Phlebotomus squamirostris, Newstead. ¢g. a, b—Antenna; c—Wing;
d—Armature ; e—Rostrum. 4, b and d to same magnification.

1 am indebted to Dr. S. Yamada, Institute of Infectious
Diseases, Imperial University of Tokyo, Japan, for the opportunity
of studying this material.

533

GLOSSINA ZIEMANNI, Grotnserc, A
SYNONYM OF GLOSSINA PALPALIS
Sus-species PMUSC TPES, Newsrrap

RaeNEWS PEA, to ReRsSi

(Received for publication 15 October, 1923)

In April, 1912, Dr. K. Griinberg* described the tsetse-fly,
Glossina siemanni, which he considered distinguishable from all
other known species by its uniformly dark colouration, its heavily
infuscated wings and its entirely black tarsi. Furthermore, by the
metallic sheen or iridescence of the scutellum and parts of the
abdomen; and by the presence also of a dirty ashen grey
pollinose covering. His description was based upon 1 ¢ and 5 9? 9
(preserved in alcohol), which were taken at Mina, Mbam R.,
Cameroons, in 1912, and forwarded to Berlin by Dr. H. Ziemann.

Being desirous of examining examples of the species in question,
| apphed to Dr. G. Enderlein, of the Berlin Zoological Museum, for
the loan of specimens of both sexes; this he very willingly granted,
and at the same time forwarded the type G and @ to this School.

On making an examination of the external characters, one saw
at once that the remarkable iridescent colouration, the ‘ pollinose’
covering and the deep blackish brown tinge of the wings were
clearly caused by impurities in the alcoholic preservative; these
impurities had so completely masked the true colours and pattern
that it was quite impossible to determine the species without making
a microscopical examination of the armature.

Having dissected. out the structures—a task of great difficulty
owing to the intense hardening of the integument—one 1s now able
to say, quite definitely, that the morphological characters are
specifically identical with those of Glossina palpalis sub-species fuscrpes,
Newst.

Sitzgsber. Ges. Naturf. xu Berlin.

*Eine neue ‘Tsetse-fliege aus Kamerun :
Jahrg. 1912, No. 4, p. 246 (April, 1912).

534

One may add that the presence of foreign matter on the wings
is easily demonstrable by passing a beam of light through the
membrane, when almost the entire surface can be seen to be more or
less covered by dark granular bodies which, in places, have blotted
out the true character of the membrane and have also matted the fine
hairs together in many parts.

One regrets exceedingly to have to relegate Dr. Griinberg’s
species to the synonymy of Glossina palpalis sub-species fuscipes,
Newst., but the study of the taxonomic characters of his material affords
convincing proof that G. zzemanni must sink.

We tender our sincere thanks to Dr. Enderlein for his great
kindness and courteous assistance, without which it would have been
quite impossible to clear up the synonymy.

535

MALIGNANT GROWTHS IN NATIVES
OPV? STO RRA LONE

BY
Se AL) ab ie
AND

E. H. TAYLOR CUMMINGS
(From the Sir Alfred Lewis Jones Research Laboratory)
(Received for publication 1 November, 1923)

It is impossible to form an opinion on the frequency of malignant
growths in natives of West Africa, as the aborigines seldom consult
medical men. Nevertheless, it has often been stated that malignant
growths are rare or absent in West Africans, and this has been
attributed to non-adoption of European habits. As a proof of the
alleged relationship between civilised habits and malignant growths,
it has been stated that such growths have only been recorded among
Creoles who have more or less adopted European habits, and in
those living in coastal towns where European influence is active.
Thus Renner (1910) states that cancerous and other malignant
erowths have been increasing among the Creoles of Sierra Leone in
recent years, but that they are rare or absent in aborigines. He
further states, that although the Fantis of the Gold Coast have been
in contact with Europeans for centuries, malignant growths are rare
or absent among them, because they have resisted the inroads of
European civilisation. Macfie (1922), on the other hand, writing
of the prevalent diseases of the Gold Coast, states :—‘ Tumours are
probably as common as elsewhere, but sarcomas appear to be rather
commoner and carcinomas are said to be rare, a belief which may be
due to the fact that the hospital clientéle represents only a small and
selected portion of the total sick.’ Dyce Sharpe (1923) states that
carcinomas are rare, differmmg in this respect from sarcomas, even
among the population of the coastal towns of West Africa. Cameron
Blair (1923) states that he has never seen a case of carcinoma or

536

sarcoma in twenty-two years in Nigeria, and that the occasional
carcinomas found by medical men in the coastal regions, occur
chiefly in natives who have come in contact with Europeans. It
must be borne in mind, however, that Europeanisation, whether or
not it were conducive to the spread of malignant growths, would
certainly be responsible for intelligent natives afflicted with them
consulting medical officers. Thus civilisation may be wrongly
blamed for the spread of the disease, when it is only responsible for
its diagnosis.

In view of the alleged rarity of malignant growths in West
African natives, the following record of seven cases, five of which
came under our personal observation between May and November,
1922, may be of interest, Five of these cases occurred in aborigines
and two in Creoles.

CASE 1. An aborigine (male Timne, aged 40 circ.) gave a
history of an ulcer on the plantar surface of the right foot following
an injury. When seen by us there was a fungating growth from the
base of a chronic ulcer, which on section proved to be a melanotic
sarcoma. ‘There was a large secondary growth in the right groin.

CASE 2. An aborigine (male Timne, aged 60 circ.) had a
painful growth on the scrotum which was found on section to be
an epithelioma.

CASE 3. A Creole (male, aged 58) had an ulcer which
commenced on the upper lip. Sections showed the ulcer to be an
epithelioma. There were secondaries in the glands of the neck on
both sides.

CASE 4. An aborigine (male Timne, aged 50 circ.) had a
tumour on the right side in the temporal region, exopthalmos of the
right eye and complete hemiplegia on the left side. Post-mortem -
Meningo-sarcoma which had destroyed a large area of bone on the
right side, involving the temporal, frontal parietal and sphenoidal
bones, infiltrated the muscles and subcutaneous tissue, and also
invaded the right orbit. There were large secondaries in the liver.

CASE 5. An aborigine (male Mandingo, aged 28 circ.) died in
the Colonial Hospital, Freetown. Dr. J. D. Dimock, W.A.M.S.,
found a tumour in the liver, which he kindly presented to the
Sir A. L. Jones Research Laboratory. Sections of the tumour
showed it to be primary carcinoma of the liver.

537

CASE 6. A Creole (female, aged 42) complained of debility.
Post-mortem: Vumour of the liver, which on section proved to be a
primary carcinoma.

CASE 7. Dr. C. H. Allan, W.A.M.S., sent a piece of liver
containing growth which he obtained from a post-mortem on an
aborigine (Sherbro). Sections showed the liver to be invaded by a
secondary carcinoma.

In addition to the above material, we examined a tumour of the
breast which Capt. M. Jackson, W.A.M.S., removed from a Mende
woman, aged 50 (circ.). The tumour on section proved to be a
carcinoma.

REFERENCES

Cameron Brarr, M. (1923). Freedom of Negro Races from Cancer. Brit. Med. fourn.,
Vol. II, pp. 130-131.

Dyce Suarpz, N. A. (1923). Freedom of Native Races from Cancer. Brit. Med. Fourn.,
Vol. II, p. 86.

Macrir, J. W. S. (1923). The prevalent diseases of the Gold Coast. Trans. Roy. Soc. Trop.
Med., Vol. XVI, pp. 156-161.

Renner, W. (1910). The spread of Cancer among the descendents of the liberated Africans
or Creoles. Annual Rep. Med. Dept. Sierra Leone, 1909, pp. 48-52.

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539

A PRELIMINARY ACCOUNT OF THE

RESULTS OF SURVEYS FOR BREEDING-

PLACES OF MOSQUITOES IN NORTH
WALES

ae Ses VW RIG EL ee Bosc:

(Received for publication 5 November, 1923)

INTRODUCTION

The present paper 1s an account of the results of an investigation
into the biology of the species of CULICIDAE, or mosquitoes, present
in various parts of North Wales. The work, which was carried out
under the Department of Zoology and the Agricultural Zoology
Laboratory of the Department of Agriculture, at University College,
Bangor, was begun in October, 1920, and has since been prosecuted
whenever circumstances have permitted; that portion of it chiefly
dealt with here, viz., observations on breeding-habits, was carried
out during the summers of 1921, 1922, and 1923.

In the summer of 1921, my attention was entirely confined to an
area of CARNARVONSHIRE bounded by the MENAI STRAITS, the
OGWEN RIVER, the SNOWDONIAN MOUNTAINS, and the GWYRFAI
RIVER. The elevation of the area worked varies from a few feet
above sea-level to about 480 feet at LAKE CWELLYN. The major
part of the district is well above the 200-feet contour ; the only low-
lying land is along the shore of FORYD BAY, an arm of the Menai
Straits. In this part there are a relatively large number of slow
running ditches and brackish pools. In the rest of the area there are
few ponds; drainage is largely by swiftly running natural streams.

The fields are for the most part bounded by stone walls without
ditches. The area is moderately wooded; the preponderating trees
are oaks and conifers; such tree-hole examination as has been done
has yielded negative results (Blacklock and Carter (1920)).

540

In the summer of 1922, and again in 1923, I re-surveyed
the North Carnarvonshire area, while I examined, in addition,
streams and ground collections of water in the neighbourhoods
of DOLGELLEY and DINAS MAWDDWY, MERIONETHSHIRE, and
ABERFFRAW, ANGLESEY (this last in 1922 only).

In all, ten species of mosquitoes have been taken in the larval
stage; viz., Anopheles maculipennis, A. bifurcatus, Aedes (Ochlero-

tatus) detritus, A. (O.) caspius, A. (O.) punctor var. meigenanus,

A. (O.) rusticus, Aedes (Lcculex) vexans, Theobaldia annulata,
Theobaldia (Culicella) morsitans, and Culex pipiens. The first
three and last four have also been obtained as imagines. The
nomenclature employed is that of F. W. Edwards (1921); since this
differs from that employed previously (é.g., Lang (1920)), I have
thought it advisable to give both the authority for the name, and
also, when Lang’s specific or generic name differs from that of
Edwards, that employed by the former author.

It may be mentioned that the weather varied during the period
which this paper covers, from being very dry and warm (1921) to
cold and wet (1923).

I have very great pleasure in taking this opportunity of acknow-
ledging my indebtedness to Professor P. J. White, F.R.S.E.,
Department of Zoology, and Mr. C. L. Walton, Adviser in Agricul-
tural Zoology at Bangor, for much valuable advice, assistance
and encouragement; to Professor Warrington Yorke, Professor R.
Newstead, F.R.S., and Miss A. M. Evans, of the Liverpool School
of Tropical Medicine for valuable assistance in the matter of
literature; to Professor Newstead, for helpful criticism also; to
Mr. F. W. Edwards, of the British Museum (Natural History), for
information as to literature, and for the identification of imagines
of Theobaldia morsitans; and finally to numerous landowners and
farmers, especially the Trustees of the Vaynol Estate and Mr. W. H.
Jones, Plas Llanfaglan, Carnarvon, for affording me every facility
for the ‘ field-work’ involved in these investigations.

Anopheles maculipennis, Meig. |
Anopheles maculipennis, Meigen. Syst. Beschr., Vol I, p. 11
(1818),

This species 1s everywhere common. In my experience, it will

r¥,

541

breed in any moderately clean water containing vegetation. The
presence of a moderate amount of current does not affect it; it will
breed in swiftly running streams, provided there is enough vegeta-
tion to shield the larvae from the full force of the current. As for
volume of water, it needs but little. In the laboratory, the larvae
may be bred in shallow vessels, offering a large surface to the air,
with low mortality; when the surface is small relative to the
volume, there is considerable mortality, even if the water be daily
artificially aerated.

According to Martini (1921), the Sanitary Staff of the German
Army in Macedonia found that this species would not breed in
water thickly covered with duckweed. I have obtained larvae from
pools thickly covered with Lemna minor. I have also obtained
larvae from water ‘ choked’ with Elodea canadense.

It is a well-known fact that in the later years of last century, and
the earlier years of this, there was a continued decrease in the
amount of endemic malaria (‘ague’) in Great Britain and also in
other parts of Northern Europe (Wesenburg-Lund, 1921). Various
theories have been put forward to account for this, the latest being
that independently proposed by Wesenburg-Lund and Roubaud,
that a change has taken place in the mode of nutrition of Azopheles
maculipennis. It has been suggested by Nuttall (Lang, 1918) that
unfavourable breeding seasons, by temporarily exterminating
Anopheles spp. in the ague zones, may have broken the ‘chain of
infection,’ and thus brought about the observed disappearance of the
disease ; he suggests that very wet seasons may bring this extermina-
tion about by washing-out larvae from the localities in which they
normally breed. I doubt if this cause would operate in such country
as is found in North Carnarvonshire, where there are few ditches,
the majority of the Anopheline breeding in isolated pools, etc., not
liable to ‘wash-outs.’ Ague was formerly endemic in parts of the
North Carnarvonshire littoral; I have met several farmers who
remember a time when it was often difficult to conduct agricultural
operations owing to ague among the farm labourers. At the present
time, there are thousands of Anophelines in the area, and many
imported cases of malaria, but so far as I am aware, there have been
no locally contracted cases of malaria within the last few years.

The observations of Macgregor (1921) and James (1922) in

542

Surrey, and my own in North Carnarvonshire, suggest that a dry
season is not likely greatly to affect the Anopheline mosquitoes.

This species generally breeds in the least shaded situations, so
that in pools or streams it may be expected on the North
side, unless this be shaded, deep, or without vegetation. I have
occasionally obtained it from quite shady spots.

On 13th September, 1922, I obtained larvae of this species from
a rainwater tank at PLAS LLANFAGLAN, near Carnarvon.

Larvae in all instar, and pupae, may be obtained throughout the
summer.

The females hibernate in the warmer farm buildings.

Anopheles bifurcatus (Linn.).
Culex oOtfurcaius, Uannaens., goyst:, Nat,, » 20. 2. oe
(1758).

This species is the commonest mosquito in North Carnarvonshire.
It breeds in the same types of water as does A. maculipennis, and
also in extremely foul pools in marshes. I have often obtained
larvae of the two species together. This species may be bred in the
laboratory under the same conditions as A. maculipennis.

Feytaud and Gendre (1919) state that, while Anopheles macult-
pennis ‘develops above all in stagnant water, clean and sunny
(clear pools, lagunes, marshes, etc.), with abundant vegetation,
variable temperature,’ etc., ‘ Axopheles bifurcatus likes pure water

. cold, with little vegetation. We especially see it in fresh
springs, streams through woods . . . wells.’ The results obtained
by Boyd (1922) and myself are not in agreement with this; Boyd
remarks that ‘ the dyke in which the greatest number (of larvae) were
found has the banks overgrown with weeds, and is almost stagnant
in parts. The bottom is covered with leaves and decaying vegeta-
tion, into which the larvae appear to burrow at times.’

Larvae may be found throughout the year in all instar, and
pupae throughout the summer.

The larvae of both species of Anophelines commonly bear a
greater or less number of ectozoic Ciliates, very similar in
appearance to Vorticella. These occur but rarely on Culicine
larvae.

The larva hibernates.

543

Lheobaldia annulata (Schrank).
Culex annulatus, Schrank. Beit. Z. Naturg., p. 97 (1.776).

This, the largest and most ornamented of our mosquitoes, is
common everywhere.

I have never found the larvae except in clean currentless water,
but it has elsewhere been found breeding in foul rainwater
(R. Newstead). It may be found in both natural and domestic
collections,

Normally, this species hibernates as the adult female, in cellars
and lofts.

Larvae of this species hibernated, in the winter of 1921-22, in
a tank at UNIVERSITY COLLEGE, BANGOR, attaining the adult
condition at the end of March, 1922. During the winter the water
in the tank was once frozen almost—if not quite—solid, while it was
frozen on the surface only, on two other accasions. Boyd (1922) has
made similar observations.

Wesenburg-Lund (1921) states that in Denmark this species is
exclusively domestic.

Lheobaldia (Culicella) morsitans (Theo.).
Culex morsitans, Theobald. Mon. Cul., Vol. II, p. 5 (1901).
Culicella morsitans (Theo.). Lang. Handbook, p. 102 (1920).

I have obtained this species from one locality only, in a wood
near GLAN-RHYD FARM, PENTIR, near Carnarvon (altitude slightly
over 350 feet), where I found numerous first and second instar
larvae in pools in a ditch, partly filled with fallen leaves and similar
debris, on 21st September, 1922. I visited this pool on several
later occasions. During the winter, fourth instar larvae were
found ; these pupated during May and June, 1923. Imagines bred
in the laboratory from larvae were sent to Mr. F. W. Edwards, and
identified as this species. After May, no larvae of this species were
found until 11th October, when I obtained a few. ‘The ditch was
full of water most of the time.

According to Wesenburg-Lund (1921), the eggs are deposited
on dry earth during July and August. These hatch out in
September, the larvae proceeding to the fourth instar during the
period ist October to tst December. The winter is passed as
fourth instar larvae.

544

Aedes (Ochlerotatus) detritus (Hal.).
Culex detritus, Haliday. Entom. Mag., Vol. I, p. 151 (1833).
Ochlerotatus detritus (Hal.). Lang. Handbook, p. 89 (1920).

This species has been obtained from the low-lying part of the
Parish of LLANFAGLAN,.bordering on FORYD BAY, and also from
similar land near ABER, Bangor, in late August, 1923.

The larvae occur both among vegetation and in open water; in
pools the large fourth instar larvae may often be seen in the clear
water towards the centre. Usually it is found in brackish or salty
water, but I have taken it from inappreciably saline water and also
from slowly running fresh water. In the laboratory, the larvae are
easy to breed; they can be kept in shallow dishes filled with water
from their original breeding-place, fresh water being occasionally
added to compensate for evaporation. I have kept larvae in two
cubic centimetres of water apiece, with no losses. By gradually
diluting the brackish water, the larvae may become accustomed to,
and thrive in, fresh water.

Under laboratory conditions, larvae pupated at various times
between 11 a.m. and 6 p.m. on 12th July, 1922, the first imago
emerging about 4 p.m. on 16th July. The maximum temperature
in the interval was 66° F., the minimum 58° F.

On 20th June, 1923, I placed some dry mud from a ditch in
which this species had bred the previous year, in a large dish of
water. On 5th July, I noticed a larva swimming in the dish. This
cast its skin on the following day, and pupated on the 11th. On
the 14th, a female emerged from the pupa. The mean temperature
was slightly over 60° F.

The imagines spend the day in the vegetation, around the
breeding-places; they may often be beaten out in large numbers.
The females are vicious biters, both in nature and under laboratory
conditions; the swelling after the bite is, in my experience, more
painful than that of any other of our North Wales species. In the
laboratory, the females will attempt to feed shortly after emerging
from the pupae, before the chitin of the mouth-parts has become
rigid enough to allow of piercing the skin.

According to Lang (1920), there are ‘at least two generations
in the year.” In 1922, I believe there were three in this area. In

545

late June and early July, and again in late August and early
September, larvae were abundant; in the interim there were none.
The major part of the second brood had attained the last larval
instar at least by 13th September, 1922, when I found only fourth
instar larvae and pupae- On 25th November, 1922, I visited several
of the pools where I had found larvae during the summer, with the
object of obtaining data regarding the hibernation of this species.
All pools and ditches were covered with ice; on breaking this and
dipping near the margin, I everywhere obtained numerous larvae.
All four instar were taken, the first two predominating. This
suggests that in the interval between my two visits (13th September
and 25th November) the females of the second brood—.e., females
hatched from the larvae and pupae of August-September—had
oviposited and that from the eggs emerged the larvae found on
25th November, these being a third generation. Larvae in all instar
continued to abound until June, 1923, when they disappeared. In
July the pools and ditches dried up. In August they again filled,
and larvae were found until 2nd October, 1923. On 31st October
no larvae could be found anywhere after a careful search. James
(1922) records that larvae were found throughout the year.

According to Wesenburg-Lund (1921) and Lang (1920), this
species hibernates as the egg; apparently it can hibernate also as
the larva. It survives drought as the egg.

Aedes (Ochlerotatus) punctor (Kirby) var. mezgenanus, Dyar.
Culex punctor, Kirby. Fauna Boreali-Amer., Zool. Ins.,
Pp. 305 (1829).
Aedes meigenanus, Dyar. Insecutor Inscitiae Mens., Vol. IX,
[res WO2T).
Ochlerotatus nemorosus (Theobald). Lang. Handbook, p. 91
(1920).

I have obtained larvae of this species from pools in a marsh near
LLANELLTYD BRIDGE, DOLGELLEY (altitude well under 50 feet), in
company with larvae of Culex pipiens, 27th July, 1922; and also
nearer DOLGELLEY in company with Aedes vexans, 27th July, 1923.

546

Aedes (Ochlerotatus) caspius (Pallas).

Culex caspius, Pallas. Reise versch. Prov. Russ. Reich, Vol. I,

p- 475 (1771).

Ochlerotatus caspius (Pallas). Lang. Handbook, p. 81 (1920).

I obtained larvae of this species in a pool on FAIRBOURNE
(South of Barmouth, Merionethshire) Golf Links, 25th July, 1922;
and with Aedes vexans, near DOLGELLEY, 27th July, 1923.
This species has previously been recorded from Merionethshire
(Tal-y-bont, North of Barmouth, see Lang, 1920) by Mr. F. W.
Edwards.

Aedes (Ochlerotatus) rusticus (Rossi).
Culex rusticus, Rossi. Fauna Etrusca, Vol. Il, p. 333 (1790).
Ochlerotatus rusticus (Rossi). Lang Handbook, p. 94 (1920).

Near DOLGELLEY, with Aedes vexans, 27th July, 1923.

Aedes (Ecculex) vexans (Meig.).
Culex vexans, Meigen. Syst. Beschr., Vol. VI, p. 241 (1830).
Ochlerotatus vexans (Meig.). Lang Handbook, p. 85 (1920).

This species has been obtained from one locality; in a field on
the right hand of the River Wnion, about half a mile below
DOLGELLEY (altitude under 50 feet).

On the evening of 22nd July, 1922, I captured a number of
imagines while beating a patch of rushes. On 24th July, a careful
search led to the discovery of the larvae, in a small ditch.

In July, 1923, I again visited DOLGELLEY, and found this pool
dry. On Monday, 22nd July, the pool was filled as a result of a
flood. On Friday, 27th July, I obtained a large number of larvae.
An attempt to breed the imagines failed, all the larvae dying,
though one pupated. Most of the skins cast were preserved and
mounted; a careful examination of this material showed it to
contain, besides Aedes vexans—in all instar save the first—Aedes
caspius, Aedes punctor metgenanus, Aedes rusticus, Culex pipiens,
and Lheobaldia annulata.

The imagines, apparently, spend the day among the vegetation.
The females attack man.

Apparently, since larvae appear soon after the pool fills with
water, it hibernates as an egg; no larvae could be found in the late
autumn of 1922.

Culex pipiens, Linn.
Culex pipiens, Linnaeus. Syst. Nat., Ed. X, p. 602 (1758).

This species 1s everywhere common. It breeds, in my experience,
in any type of water, natural or domestic. I have obtained the
larvae from streams, ponds, pools in marshes, water in hoof marks,
and all sorts of rainwater receptacles. It will breed in very foul
situations. It is easy to breed in the laboratory.

The female hibernates in dark, cool cellars or lofts.

It rarely, if ever, attacks man. I have never succeeded in getting
it to bite under experimental conditions.

REFERENCES

Bracktock, B., and Carter, H. F. (1920). On the Results Obtained from Surveys for
Breeding-Places of Tree-Hole Mosquitoes in Liverpool and Neighbourhood. <Axn. Trop.
Med. & Parasit., Vol. XIV, p. 115.

Boyp, J. E. M. (1922). The Botany and Natural History of the Dyke-land near Sandwich,
Kent, as far as they concern Medical Entomology. ournal R.A.M.C., Vol. XXXVIII,
pp. 41 and 117.

Epwarps, F. W. (1922). A Revision of the Mosquitoes of the Palaearctic Region. Bull. Ent.
Research, Vol. XII, p. 263.

Frytaup, J., and Genorg, E. (1919 i). Sur la Répartition des Gites d’ Anopheles maculipennts
et d’ A. bifurcatus. Bull. Soc. Path. Exot., Vol. XII. 1919.

(1919 ii). Sur la Répartition des Gites d’ Anopheles et d’ A. bifurcatus. Procés-Verb.
Soc. Linn. Bordeaux, Vol. LXXXI. p. 24 (Cited by Boyd. Summary in Rev. App. Ent.,
B. Vol. VIII, p. 211).

James, S. P. (19221). Mosquito Life in Surrey during 1921. Bull. Ent. Research, Vol. XII,
p- 427.

(1922 ii). Notes on the Eggs of Culicine Mosquitoes found in England. Trans. R. Soc.
Trop. Med. Hyg., Vol. XVI, Dec. 1922.

Lanc, W. D. (1918). A Map of the Known Distribution in England and Wales of the
Anopheline Mosquitoes. British Museum (Natural History), London. 1918.

(1920). A Handbook of the British Mosquito. British Museum (Natural History),
London. 1920.

MacGrecor, M. E. (1921). The Influence of Drought on Mosquito Life in Surrey. Bull.
Ent. Research, Vol. XII, p. 205.

Maartint, E. (1921). Die biologische Malariabekampfung in Mazedonien. Leitschr. angew.
Ent. Berlin, Vol. VU, p. 225. (Summary in Rev. App. Ent. B. Vol. IX, p. 91).
WesEnBurG-Lunp, A. (1921). | Contributions to the Biology of the Danish Culicidae. D. Kgl.
Danske. Vidensk. Selsk. Skrifter Nat. Math. Afd., 8 Raekke, Vol. VU, p. 1. 1921.

*
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549

THE FREQUENCY OF INDICANURIA

BY:

R. M. GORDON

(Recewed for publication November 6, 1923)

TEST EMPLOYED

To five ccs. of urine in a test tube were added one large drop of five per
cent. potassium chlorate, then five ccs. of strong hydrochloric acid,
followed by five ccs. of chloroform, the contents being mixed by inverting
the closed test tube a couple of times. If no definite blue colouration
appeared in the separated chloroform after thirty minutes, the result
was regarded as negative.

RESULTS OBTAINED

Three hundred and eighty cases were examined on one occasion
each; of these, ninety were apparently healthy, normal individuals,
twenty-two (24 per cent.) of whom gave positive results and sixty-eight
(76 per cent.) negative. It is to be noted in this connection, that the
same individual may give different results on various dates, or even at
different hours on the same day. Thus a normal case tested on twenty
occasions, was negative on twelve and positive on eight, a negative result
frequently alternating with a positive. The remaining two hundred and
ninety cases were undergoing treatment for various disorders. The
results of the tests are shown in the table on page 550.

REMARKS

The presence of indicanuria is usually attributed to excessive putre-
factive changes in the intestine (Emerson (1913), Hawk (1919), Cole (1920),
Heitzmann (1921)). Its presence in sprue cases is remarked on by several
authorities. Thus Cammidge (1912) found it present in eighty-five per
cent. of his cases; Rademaker (1906) mentions it as a diagnostic point,
while Castellani and Chalmers (1919) and Byam and Archibald (1923)
both refer to its presence ; Bahr (1915) notes its occurrence in some of his
cases, but regards its presence as of no great significance. ‘Three out of

TABLE

Showing the frequency of indicanuria amongst three hundred and eighty individuals

Number Percentage

examined positive
Normal individuals ore Af oes He $.¢ ics aes go 24
Sprue oa re ait bis oat esi us ae da 4 75

Amoebic Dysentery = rr lS las 7 yeaa rad hf etal: 4 35 94
Bacillary Dysentery Feel bs inkeh ants hq eee ca eae ets 5 °
Diseases of stomach and small intestine other than the above... 6 17
Diseases of large intestine other than the above... a a 23 56
Diseases of the genito-urinary tract ... Red bes “th Yr 17 30
Surgical conditions other than the above ... 0... 0... of 65 15
Lung cases... one ae ae nae ate ee <3 i 62 40
Malariasgeeee- rae mas Pe any see aie ay ie 20 50

Various... ee ies <i fee ean ae 7 — 53 27

551

the four cases examined by the present writer were positive. It will be
seen from the table that the highest percentage of positive results (94 per
cent.) was obtained from amoebic dysentery patients, while the five
bacillary dysentery cases examined were all negative. Obviously the
number of bacillary cases examined is too small to allow of definite
conclusions being drawn, but the marked disparity between the two would
suggest that the test may be of some value for differential diagnosis.
Quincke and Roos (1893) have drawn attention to the constant presence
of indican in the urine of two amoebic dysentery cases, observed by
them for respectively eight and eleven months. Ten out of twenty cases
of malaria examined gave positive results. The increase of indican in this
disease has already been remarked upon by Marchiafava and Bignami
(1900) and Craig (1909).

CONCLUSIONS

Indicanuria occurs in about twenty-five per cent. of apparently
normal individuals.

It was present in ninety-four per cent. of amoebic dysentery cases,
but was absent from the urine of five cases of bacillary dysentery.

REFERENCES

Baur, P. H. (1915). A Report on Researches on Sprue in Ceylon. Res. Mem. Lond. School Trop.
Med. Vol. Il, p. 58.

Byam, W., and ArcurpaLp, R. G. (1923). The Practice of Medicine in the Tropics. Vol. III,
p- 2256.

Camminetr, P. J. (1912). Sprue: Its Diagnosis and Treatment (by Begg, C.), p. 49.

CasTELLANl, A., and Cuatmers, A. J. (1919). Manual of Tropical Medicine. 3rd Edition, p. 1787.
Corr, S. W. (1920). Practical Physiological Chemistry. 6th Edition, p. 318.

Craic, C. F. (1909). The Malarial Fevers, p. 142.

Emerson, C. P. (1913). Clinical Diagnosis. 4th Edition, p. 152.

Hawk, P. B. (1919). Practical Physiological Chemistry. 6th Edition, p. 152.

HerrzMann, L. (1921). Urinary Analysis. 4th Edition, p. 77.

Marcuiarava, A., and BicnaMt, A. (1900). Twentieth Century Practice. Vol. XIX, p. 226.

Quincxz, H., and Roos, E. (1893). Ueber Amében-Enteritis. Berlin. Klin. Wochenschr. Vol.
XXX, p. 1089.

Rapemaker, G. A. (1906). Onderzoekingen naar aanleiding van een Geval van Indische Spruw.
Dissertation. Leiden.

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553

MISCELLANEA

NOTES ON CESTODE PARASITES FROM A
DUGK

©np Julyisth,! 1922) CAs 2 W2 PiNoel EPullers, \ Fusq.5. FLRIG.V.S.,
obtained from York the intestine of a duck, which was found to
contain about 300 Cestode parasites.

As the gut had been removed about two days previously, the
parasites were partially moribund. The gut was slit open and
placed entire in hot Schaudin’s preserving fluid, and later, some
hundreds of parasites were removed. They were identified as
under :—

(1) Hymenolepis megalops (Nitszch, 1829), Par., 1899.

About 20 specimens.

(2) Hymenolepis coronula (Duj., 1845), Cohn, 1901.

About half the collection consisted of this species. The
rostellum was armed with about twenty hooks, measuring
from 13m to 17m, of the shape figured by Lie.

(3) Aploparaksis furcigera (Nitzsch, 1819), Fuhrmann, 1908.

Over one hundred specimens were obtained. Micro-
scopical examination of stained specimens revealed the
fact that, whilst nearly all segments contained one testis
only, other segments contained two, whilst testes were
entirely absent from other segments. [his phenomenon

was noticed in seven or eight strobilae.
T. SOUTHWELL.

NOTES ON PARASITIC WORMS FROM THE
GOLD COAST

Dr. J. W. S. Macfie sent a number of specimens from Accra,
Gold Coast, West Africa, which were identified as follows :—
(1) Davainea tetragona (Molin, 1858), R. Blanchard, 1891.

A large number of specimens from hens.

554

(2) A coenurus from the jaws of Mus rattus, and also from

the pleural cavity of Crzcetomys gambianus.
This larval form was first described by Turner (1919). The
adult form is not known; the hooks bear a close resemblance to those

of C. cerebralis and of C. serzalis.
T. SOUTHWELL.

CITTOTAENIA LAGORCHESTIS, Lewis, 1914

This worm was obtained by Dr. Maplestone from the stomach
of an agile wallaby (A/acropus agilis), taken near Townsville, North

Oueensland.
T. SOUTHWELL.

DAVAINEA LEPTOTRACHELA, Hovne., 1910

A complete cestode worm from the small intestine of 7u7zdus semz-
torquata (Turtle dove), Pietermaritzburg, Natal, was collected and
presented to the School by Mr. Hill, Pietermaritzburg. It proved
to be a specimen of the above species.

The suckers are armed and the genital pores irregularly alternate;
ovary asymmetrical, situated slightly on the pore side; three or four
eggs per capsule, the capsules extending in posterior segments to
the lateral margins.

Hungerbiihler recorded this species from Péeroclidurus namaquus

(Grouse).
T. SOUTHWELL.

PARAMPHISTOMUM CERVI IN A HORSE

Some worms collected in May, 1920, from a horse at Tamale,
Northern Territories, Gold Coast, and kindly sent to us by
Dr. K. B. Allan, proved to be Pavamphistomum cervi. This record
is of interest because, so far as we are able to ascertain, this parasite
has not previously been obtained from horses, and is not mentioned
as occurring in this host by Maplestone in the ‘ List of Amphistomes
arranged under their hosts’ appended to his recent revision of the
Amphistomata of Mammals. J. W. S. MACFIE.

555
A NOTE ON AUCHMEROMYIA LUTEOLA, Fas.

The bionomics of this fly and its larva the Congo Floor Maggot,
first described by Dutton, Todd and Christy (1904), have been very
fully described in recent years by Roubaud (1914).

Fic. 1. Auchmeromyia luteola. Larva feeding on human skin. =X Io circ.

The photograph shows the method of feeding which the larva
adopts. It stands more or less at right angles to the skin, and has
such a firm hold that when the limb is turned over it goes on
feeding in a hanging position with equal facility. The feed lasts
for as much as an hour in many cases.

The larvae, of which one is photographed feeding on the human
arm, were brought alive to England from Sierra Leone at room
temperature in sand.

B. BLACKLOCK.

ANCYLOSTOMUM CEYLANICUM IN CATS
AND DOGS OF SOUTH INDIA

Ancylostoma caninum and A. ceylanicum were found in all of
five dogs examined at the Veterinary College, Vepery, Madras; in

the single cat examined only A. ceylanicum was found.
L. S. PARAMESWARA AYYAR.

THE URINE IN

556

MALARIA

Nephritis as a concomitant of malignant tertian malaria is referred to

by most authorities, but its appearance in quartan and simple tertian

seems less well known.

The following is a record of sixteen consecutive

cases of malaria examined at the Liverpool School of Tropical Medicine.

Number Number
Parasite of cases of cases
examined positive
P. malariae I I
P. vivax 2 z
P. falciparum 13 7
Protocors or Positive Cases*
Number | Duration Deposit in 5 c.c.’s of
of of Quinine | Temperature Parasite Albumin centrifuged urine
case attack

I | 4 days Yes 103° P. malariae + A few granular casts and renal

| cells.

2 | 3 weeks Yes 105° PB. vivax + A few casts and renal cells.

3 | 3 weeks Yes 98° | P. vivax fe) A few renal and red cells.

4 3 weeks Yes 98° P. falciparum + A few hyaline casts and renal
cells.

5 2 weeks Yes P. falciparum ++ Large numbers of granular and
hyaline casts and a few renal
and red cells.

6 2 months | Yes 98° P. falciparum + Nil.

7 2 months | Yes 98° P. falciparum + A few renal and red cells.

8 I month Yes 103° P. falciparum fe) A few renal and red cells.

9 2 weeks Yes 98° P. falciparum fo) Many renal cells ; a few red cells.

10 3 weeks No 98° P. falciparum + A few casts and renal cells.

* By ‘ positive’ is meant the occurrence of any, or any combination, of the following, albumin,
casts, renal epithelium.

R. M. Gorpon.

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PAGE
HSE Saat aE Ea CoP RR aN Tae epee ERY SOU mean 2 See, Pa ll
CSPMIEBASP LN DEX Gs: Shy dove nso cnnns dt ohescchass 02 iE LION ead iil
InpEx oF GENERA, SPECIES AND VARIETIES NEW TO SCIENCE .....e.see0e viii
INDEX OF AUTHORS
PAGE PAGE
SURES ey er ener ee 13, 427 AV Pia orl Bias spreern 5, 9 439, 440, 554
mdier, >.> and Clarke. KE. J........... 299, 439. Mactcvsla We go; and workes \W....e0.;. 439
Adler, S.; and Cummings, E. H. Taylor 535 ey BIg aa Yop) eget 2g “patina pet met LW, he
paler, 2 and southwell, Us .......+++s. ga Maplestone, P. A.; and Southwell, T. 317
BORE La re ATAINESWAIA. 0. vekwecssecescacs 555 Ie syaLOACl WncMeer es eetenta cert mecca ace etn ce
BARRE OCR Diy ref Bins oi ot nats ann 0) 79¢ 337.555 Carat. Crale ali; aud Uole. |. te W aee 003i
Blacklock, B. ; and Thompson, M. G.... 443 OTe ete et Veta cctra ss esac tee reeck ras ree 93
BP PISLOD UCTS ea ht tis ote cn aossdte ote) 7a pelWwyo-Ciaike. bo oe. ew anu, Gr. lis
Petco ers Las and. Adler, 9. ica. cess 299, 439 fala ly VST atin er TR gan eR ees are ee: 389
Corson, [VSL oa aR eran en BN SOM RIVE tem Linen sacene date ecu ar ets eet!
Cummings, E. H. Taylor ; and Adler, S. oe Southwell, T’..; and Adler, S.°7..°........ 224
Te TG LE a 6 Oreo Serer Sil Southwell, T. ; and Maplestone, P. A.... 317
(RIE EOC tt RnR Ta a Aan 19 BEC Miredo. os Wien VV nomena crc eececatan poe 303
oo TETAS Ce 89, IOI, 377 UAL Str tee ct arena eee ee eats eee 359
BOOT eine Wisc. cs nesses 289, 381, 549, 556 Thompson, M. G.; and Blacklock, B.... 443
a eas 22s he se sp nenr Si GE ont 347 Wenvole Cr Nite auriecccenors semen et tae? 231
thle, J..WaW.z and Oordt,.G, J. Van:... 31 AEG On EG 2 ane gameplan tage ye = cnc Satn a 431
Ingram, A.; Selwyn-Clarke, P. S.; and NER? UG Wie CCS a don. ote eee teer crs 539
AST) New oo asin ath 6 Sin'n tg bon G0 389 MOTEG. a VV cuatiUalv ace, -[aa\Wq wee seca te 439
re A RI os os oe cals ecg Sans ne 47, 59
Le Fanu, G. H.; Selwyn-Clarke, P. S. ;
and Ingram, eo ee " 389
GENERAL INDEX
PAGE PAGE
Adler, S. Carbon Tetrachloride in the Adler, S., and Southwell, T. A Note
sLreatiment, Ol Pilaryasis |... feces d+ s0: 427 on Opbhiotaenia punica (Cholodovski,
Adler, S. Malaria in Chimpanzees in FooS): Lg. Riles nt OL Te, 54.95 nisi 333
Bierrd scOnC ilmageuted. «i eeuycnasnavaya+ ves 13 Aedes (Ecculex) vexans (Meig.), Breeding
Adler, S., and Clarke,,E. J. A case of Habits: Ol ae naare ian eae ee ees 546
acute ascending Paralysis in a » (Ochlerotatus) casptus (Pallas),
Whats panzecmertesilegas to -. +060 .re0sye%s 3 Breeding. Ela pifs Olea ts gts 546
Adler, §., and Clarke, E. J. Kurloff 55 (Ochlerotatus) detritus (Hal.),
Bodies ikebash. fest hola ones 0. des ieee, 5800 439 Breeding Habits’ oft uc antkerr 544
Adler, S$., and Cummings, E. H. Taylor. »5 (Ochlerotatus) punctor (Kirby) var.
Malignant Growths in Natives of meigenanus, Dyar., Breeding
PLETE Am CONGE ae - Gi tdgd can: rorcss cogs 535 Phe es, ha ae 545

ill

PAGE
Aédinus amazonensis, LUtZ..........c000008s 377
Amoebic Dysentery, Treatment of ...... 381

Amphistomata of Mammals, Revision of 113

Amphistomes, arranged under their Hosts 202
Ancylostomum ceylanicum in Cats and

Dogs of South India
Anopheles amazonicus, New Species of
Myzorbynchus Group from

pouth Americaaaas ne ace 71

LEE

Coe eeeeerereeeeses

és, annulipes, Distribution in
Australia‘ota. = eee 222
% bancrofti, | Occurrence in
Austrolia hers 224
» bifurcatus (Linn.), Breeding
tlabipscote et nia eee. 542
+ costalis, Action of Lithium
Chloride on Larvae of ...... fe)
i maculipennis, Meig., Breeding
Vic Vey BAche 6) Saute ie Sea 540
. ALLOUTOSSCRS 184s cesaenv eugene 75
x (Arribalzagia) punctimacula... 101
(Arribalzagia) venezuelae, syno-
nymous with Anopheles

(Arribalzagia) punctimacula 101
Anopheline Mosquitoes found in Australia 222

Auchmeromytia luteola, Fab. ............... 555
Australia; Malaria iniecy a eee 213
Ayyar, L. S. Parameswara. Ancylos-
tomum ceylanicum in Cats and Dogs of
south India tec. semen ee ee 555
Balanorchis, actus en ee 165
Belgian Congo, A new species and a new
Variety. of Gules oni ee ee 89
Blacklock, B. The Etiology of Black-
water Feverct ct, ee ee 79
Blacklock, B. A Note on 4 uchmeromyta
lutcola;/Fabse Bice te ee 2, ee 555
Blacklock, B. A Pyrrhocorid Bug
capable of bignesiVianeas. eee 337
Blacklock, B., and Thompson, M. G.,
A Study of the Tumbu-Fly, C ordylobia
anthropophaga Griinberg, in Sierra
Leone*’ .27240 Se, ee Oe ee 443
Blackwater Fever Blood, experimental
Inoculation" ot ae a 84
Blackwater Fever, the Etiology of ...... 79
Bothridium ornatum, n.sp. .........0eeeeee B17
Brumptia, Genus ))2:. 1, ee 196
Brumptia' gigas: “2 HS Bea 196
Carbon tetrachloride in the Treatment
of Milariasis 204, 1 eee 427
Carmyerias, Genus’ .2.....20, eee. 178

PAGE

Carmyerius cruciformis ..cccccecccssceescens 181

ra EXOPOTUS, TLBD..>csosaverereniie 183

~ BVCRATIUS ... coerce eee eee 183

33 SPALLOSUS” cvweyeauvses esas eaten 179

Be WOENYONL) “Nh trates 1 181

Cats, Coccidicsis‘of “Jig:..veeeesses gp cane Z3t
Ceratopogonine Midges, Species attacking
adult Lepi-

dopteray ...... 22
“ - Species attacking

caterpillars ... sae
Species attacking

99 29 A

Mosquitoes 2

. He Species attacking
Oil-Beetles*.:) ae

“ b Species attacking
a Phasmid ... 28

- : Species attacking
a -Oldlin woe 25
Cestode Parasites from a Duck............ 553
Cestodes from Casuariformes............... 47
» : Galliformes..............004. 59

Chimpanzee, Case of acute ascending
Paralysis ijt t op ee 299

Chimpanzees, Malaria in Sierra Leone in 13

Chrorchis, Gettoe 0-73 Cpt. ys 3s, 165
Cittotaenia largochestris, Lewis, 1914 ... 554
Cladorchinae, Sub-family .............0..05 158
Cladorcthis, Genus cee 166

Clarke, E. J., and Adler, S. A Case of

acute, ascending Paralysis in a Chim-

PalZee Mier cht iaevccmes > eee ee 299
Coccidiosis of Cats and Dogs and the
Status of the Isospora of Man ......... 231
Coccidium bigeminum, vars. canis, catt,
PULOTT. Fre saccet res +s nce hee eee 239
Cordylobia anthropophaga Griinberg...... 443
C. anthropophaga, Animal Hosts in
Nature of (ice 481
s Comparison with other
Myiasis-producing
Flies.t.22 ae 499
3 Development in
Animals 220). ae 479
5 Kedyses*of O08, 0c.) 477
# Ege of URUk.5,. ae 456
53 Fertilisation of the
Female. 8, .feiat 478
‘3 Food: of -adultsti 720% 451
i Geographical Distri-
bution tof. AVG 447
43 Larva-of 20m. AR 457

PAGE
C. anthropophaga, Mode of Infection of
M

ier bade pty? 494
se Morphology and Bio-
eC ee Ol ae renee 449
he Oviposition of ...... 452
= Pathogenicity to
iTV ittra Sepeede resect 479
= Datei Chants. <er 472
_ Seasonal incidence of 492
xy Size OF AUUIE tre, ccs 478
by Infection, Main natural
Reservoir of ...... 482
5 Infection, Prophylaxis 498
= Infection, Relative
Immunity of older
aVitiia iSeries teeter et 488
te Infection, Symptomat-
oy ahes 0 yeaa ere bh 495
he Infection, Treatment
he ie oe messes 497

Corson, J. F. A case of Creeping Erup-

tion in a European in the Gold

WOE terre atten aaa area tar Rota I

Mer ODDOTUnECTENUS Merit tests tsk ceetee tes I51

hs BOL VEOPUOTUML on wencateus sees ISI

3 PETHOPRILE Bivacoe creme rete te 158

= iA alte ihe Peas HIE ahh 4 158
Creeping Eruption in a European in the

ei A BT gale | pre cia op ea eR E ED I
Culex, A new species and a new variety

from the Belgian Congo ...... 89

55 annulioris var. congolensis, N. var.... QI

,, jatigans, Action of Lithium
Chloride on Larvae ote... as IO
5» maracayensis, n.sp. from Venezuela 102
hy EEG ETS TRS eee 4 Yee pipet yeti 89
5) PAANIUS, NEW SPECIES... .400seeeeeeeees 104

pipiens, Linn., Breeding Habits of 547
Cummings, Ear: Taylor, and Adler, S.
Malignant Growths in Natives of

EE ete Meta vocets. saranda core et > 535
Cyclicostomum Larvae in the Horse ...... 32
Davainea appendiculata .........cccecececees 49

f (s.l.) casuarit, new species of
PUVIaie WC estUdems cere ees 47
os (Sy LENO PUPDIIEES errs fete 49
ca geal hi cep tippetie’ betel tee at. 49
" Ce yyrarreguesiic Tl-SPrb t.cccee 55
» leptotrachela, Hung., 1910 ... 554
RITEPS Se DOPUAIOT AN COs an eer rere sete ae tiee 67
Bee TECDISPDAN MS, LSD waeett resect acre: 63
“? WRT TA TBE). etss tes deractaeces teter® 59

PAGE
DoereCocerdiosis "Or Sit i. aisen. nithas anes 231
Duke, H. Lyndhurst. ‘The ‘Transmission
of I. congolense by Glossina palpalis... 511
Dosdet cus) BiOnOnu Ce Of een, st eerteu vee 339
Gee IP SUDETSTATIOSHS esti cs teten. ek ho 37
53 e F., Flagellate
Parasite! of 7) 442
Edwards, F. W. New and Old Observa-
tions on Ceratopogonine Midges
attachilry Other iisectswrrd.t fsscds5s.00% 1g
HEMCT UI EOTASY TSP yi leis cy'suitnn etree 261
Emetine periodide, Treatment of
amoebic Dysentery with ............... 382
Evans, A. M. A new Species and a new
Variety of Culex from the Belgian
COPRORE, Pre stestir ete sete tt chs o tates thes 89
Evans, A. M. Note on 4édinus amazon-
PL Mek pyih AS wghan oe ee ia gp) aS Na a 377
Evans, A. M. Notes on Culicidae in
Venezuela, with descriptions of New
SDECIES) seat to rate terse. 5 fEce bas one ee 101
Filariasis, Carbon tetrachloride in the
MeALMeNt Oka sarc cceten trees as tees 427
Perchoederius, Genussasese. dc Atty «1x 176
- CODDOLAL credeces OTTO vee ne 178
rs; GIS CDOLUETE rico tc TR eel sess 177
Forcipomyta alboclavata ..........cceceeeeees 26
. CPUDEIES © heed du ercdeeent eke 26
" TN le ce A ARE 26
ee (Euforcipomyia) papilionivora,
SPsD soar Acee A aeedcdsddnetet ete 23
GrasiPoalsctaaes DAMiny soe, Rene fas 5 sbees ste 187
Gastrodticpides, (seus eee cate tas. sce e eee 193
is POURS tar 0 oF oot os ttc tote e 194
Gastrodiseus, sens. ccc ear ee 187
Toi BEOMPLIBCUS 2403.0. Bie eee Igo
. MINOT As ee acan vtec. Seep e Te ets ee 193
. SCOUNAUS?2T3i. ccs lee eee 193
Gastrothylacidae, Family ..............068. 169
Castrortpylie( Sentis tio. ts, sodvasoteees covert: 169
7 bubalts* riciiliii. teens 182
CTUMENS ENO. thd tant ee 175
Glossina palpalis, The ‘Transmission of
PCONZOLERSE DY NG. tiawk tee Sil
ziemannt, Grinberg, a synonym
of Glossina palpalis, sub-
species fuscipes, Newstead ... 533
Goeldia longipes, specimens from Vene-
AV AE he Alea te ns or ech eer re Rel a oe 18 fe)
Gold Coast, Parasitic Worms from ...... 553
ma re Relapsing Fever in ......... 389
£ a Yellow Fever in............... 431

PAGE

Gordon, R. M. ‘The Frequency of
Indicanuria ext 4tE. jer dipect-cbees
Gordon, R. M. A further Note on the

Occurrence of Ancylostomes resembling
Necator americanus amongst domestic

Pigs snaAimgao nate a= cee oneersesg cre 289
Gordon, R. M. ‘The Treatment of
amoebic dysenteriae sprays -- cues deny 381
Gordon, R. M. The Urine in Malaria 556
Hehir, P. Typhus Fever in Greek
Refugees, ).crersesa ress scstteeges a8 Aa sees 347
FHlomalogastarsGsenus” ..capkrtieprwces.. -apeh 195
ee DOLONGA bing: 54 ophteree CS Tobey 195
Horse, Strongylid Larvae in............... 31
HLouttuynta Struthionts ....cccocecscsecceeeees

thie, J. EesWee wand (OordtsaG.5 | aan.
On some Strongylid Larvae in the
Horse, especially those of Cyyclicostomum 31

Indicanuria, The Frequency of............ 549
Ingram, A.; Selwyn-Clarke, P. S., and
Le Fanu, G. H. Relapsing Fever in
the Gold Gigast bares pyaar: sprees? 389
Lioshora Dig¢mindies. sc. «tee 257
J sc uagl CHS US Digeess earet hans 248
* of Manso. 2.. Senet eae 221,263
ra FEU MEET OORT. cs 81 Ee, CRORE 260
Jaundice, in Malaria and Blackwater

Fevers. i156... casei es eee ee 82
Kotlan, A. Avian Cestodes from New

Guinea. II. Cestodes from Casuari-
LOLINGS 52a telethon cck yaaa 47
Kotlan, A. Avian Cestodes from New
Guinea. III. Cestodes from Galli-
FOTMES | 7h... ce ate, Ree. eee Aen ene 59
Mourlott. Bodies in Fish: aasatn pee 439
Le Fanu, G. H.; Selwyn-Clarke, P. S.,
and Ingram, A. Relapsing Fever in
thesGoldiCoasty 2 seesicst...aeerer 389
deustomia Cchidnae SH ® : i635. s ede Macs 326
Lithium Chloride, Action on Mosquito
Latvael of sfx. Fossey eee te eee 9

Macfie, J. W. S. Depth, and the Larvae

and Pupae of Stegomyta fasciata, F.... 5
Macfie, J. W. S. A Note on the Action

of Lithium Chloride on Mosquito

Laryac. ., textarea eee ees cae 9
Macfie, J. W. S. Onchocerca armillata in
Cattle in the Gold sCoast.ssunel oe 440
Macfie, J. W. SS. Paramphistomum cervi
anja Tlorse.ss esas thf: opkee quem mney? 554
Macfie, J. W. S. Pigs and Ankylos-
comiasis in theyGold ‘Coaste.@....:7... 439

vi-

PAGE

Macfie, J. W. S., and Yorke, W. I7i-

chonema tetracanthum, Mehlis, 1831,
Of T08%, (LOO sabe Teerot ines. cotese ae 439

Malaria, Effect of Season on ‘Treatment
9) Sey ea FS = EE ee ee: 307
Haemogiohbinurigin:.......c.0.00 81
sieht Australias Atton ct ee eee 213

6 in Chimpanzees in Sierra
TeCOnE fever eee 13
7 Insect vector in Australia...... 225
e Studies in the Treatment of... 303
Urine 209-5 nfs eee 556

Maplestone, P, A. Malaria in Australia 213

Maplestone, P. A. <A Revision of the
Amphistomata of Mammals .............. 113

Maplestone, P. A., and Southwell, 'T.

Notes on Australian Cestodes............ 317
Megarhinus trinidadensis specimens from
Venezuelan ci! ois. epesckech arte eee 110
Wliscellatiea..ic¢.s:-42checbes See eee 439, 553
Monopylidium freldingt, sp. NOV. ........... 318
MACTACANLHUM...... 0. .eeeeees 322
Mosquito Larvae, Action of Lithium
Mhloride on) 2.10201.0.20; cae oe 9
Myiasis due to C. anthropophaga ......... 444
Myzorhynchus Group. Anopheles ama-
Zonicus, sp.n. from South America..... 7%
Necator americanus, Ankylostomes resem-
bling. j:vcieascee nee 289
$5 2 Attempts to infect
Pigecwith 8 oes 296
Newstead, R. Glossina ziemannt, Griin-
berg, a Synonym of Glossina palpalis
sub-species fuscipes, Newstead ......... 533
Newstead, R. On a new Species of
Phlebotomus from |apan,., ...sccatbasnenes? 531
North Wales, Breeding-places of Mos-
CUALORS TILT. «-eorllepaaf8in topmye So Eicg seat amas 539
Onchocerca armillata in Cattle in the
Gold Cogst eer. oe nu bees tei tee eee 440
Oordt, G. J. van, and Ihle, J. E. W. On
some Strongylid Larvae in the Horse,
especially those of Cyclicostomum ...... 31
Opbtatagns a DUNC. Shc dn etc eee 333

Orpen, R. W. Report on Sleeping Sick-
ness in Eket District, Southern Nigeria 93
Paralysis, Acute ascending, in a Chim-

Panzee face ss es Oe 299
PAS AMONIETIG Sig Sy TIP TS) ieee tes nce 329
Paramphistomidae, Family ..............244- 1s
Paramphistominae, Sub-family ............ 116
Paramphistomum, Genus ..........cceeeeeeee 116

PAGE
Paramphistomum, Summary of chief
characters of six
BHEEIES OL seer 128
He PMAATIORS 3 oc tes As ketaese 150
calicophorum .....0..000 139
a COTUR TOMEI L, TEs Wedeass 117
~ we Nina ELOTSG?, tect) 554
A CLASIME rete peat 141
aA dicranocoelium .......... 146
- EXPLANALUM........e.0e0e 137
" gigantocotyle... ... ...... 133
< HOTCDES. Pi aces soks rate 132
be OTEDOCOCILUM .....c00e0008 145
Ne parvipapillatum ........ 147
nN PES en ccrnnon asutal et o- 132
scolrocoelium ........000s 147
+ SHIPLEY Ur sict ony on cee ts ee 148
s SURTONICLS ch bth ei 143
e streptocoelium — ......... 146
“ TA GEL UES ON ee CER EY 150
OEE DOGAG TAILS meceryy aire jarpe ee Sena gie 165
Phlebotomus squamirostris, N.SP....3..00+00+5 531

Pigs and Ankylostomiasts in the Gold

CHET Eta asi co ease Oe ane tre aE 439

MteucOU SS CUS” GrONlIS; 0225 rach os oath ne dacyoaans 160

" POLUH Str tee es eee es 160

= VCO RES Tisai seat a Sree 164

DL IPLOV IEE, Tite cay aoe elon 161

= OOO ata rtion cone th Haan? 164
Psorophora ciliata, specimens from

TIO CElAMe ee Mai viee. i255 108

¥ ad gd oy SRY MPEP OTTER ELLOS 106

Pyrrhocorid Bug capable of biting man... 337

Quinine Haemoglobinuria compared with

Biaccwater: bever...) vg .e 80
Pee Via cer OSE Off, os csarrces cae e 308
Raillietina (Parontella) paradisea ......... 49
op (Ransomia) campanulata —_ 49, 56

Bs “ RAUIATE Sie ce-penes 49 ,
, 3 DALANGA few cciuse es 49
as (Skrjabinta) oligacantha ...... 49
Relapsing Fever in the Gold Coast...... 389
», Animal Experiments ... 406
_ », Clinical Manifestations 408
> Sun Did en IOlOp yee veer a, 389
“a SEAT UT Cyaeedecesee ees 419
= Pa Ificiccnice...s.001 eee a 390
fe », Preventive Measures ... 422
“ ope INCE Sa) Ciel leper geoe 414
. py CCALIOCI Liye. t en,’ 20

Revision of the <Amphistomata of

UN ET cae Eta aan an eee ere 113

vii

PAGE
Sachs-Georgi Reaction, Certain fallacies

1 OS EM oo Se Oe ae a Sea Se 366
Selwyn-Clarke, P. S., Le Fanu, G. H.,
and Ingram, A. Relapsing Fever in
tlre Grolds Coasteiad i082. sbcdee sia da meds 389
Sierra Leone, Malaria in Chimpanzees
VN PAVE keno 13
‘3 Occurrence of C. anthropo-
PpPaga lms Wd ascieeee ee 448
Sleeping Sickness in Eket District,
Southern Nigeria, Report on ........... 93
Southern Nigeria, Report on Sleeping
Bicknessitr Kets IStliCh wscuge ater reise 93
Southwell, T. Cuttotaenia largochestris,
WeWsiSalOTAG tes cetecdees cote aciyler sts hs tas 554
Southwell, T. Davainea leptotrachela,
Pilino spl OLO ete eet tie 554
Southwell, T. Notes on Cestode
arastvecuronivanl SUCK en. vets: accee es 553
Southwell, T. Notes on Parasitic Worms
trom therGold Coast... po. k hte daet « 553
Southwell, T., and Adler, S. A Note on
Ophiotaenia punica (Cholodovski,
F003) hic 1O) (eee re ees: 233
Southwell, T., and Maplestone, P. A.
Notes on Australian Cestodes ......... 317
Stegomyia fasciata, Action of Lithium
Chiorideroin latys car meer. rata. 9
Stephanopharynginae, Sub-family ......... 167
Stephanopharynx, Genus ......eeeeeeee eee 167
COMPALTUS ho ds sece eh cae 167
Stephens, J. W. W. Studies in the Treat-
Ment ob vialariawre Aneta, tee ee 303

Strongylid Larvae in the Horse............ 31
Stuart, G. The Value of the Sachs-
Georgi Reaction in the Serological
Diagnosis of Syphilis
Syphilis, Value of Sachs-Georgi Reaction
in the serological Diagnosis of
Taxorchis, gen. nov.
Theobaldia annulata (Schrank), Breeding
Habits of
os (Culicella) morsitans (Theo.),
Breeding. Habits off. eee ee
Thompson, M. G., and Blacklock, B.
A Study of the Tumbu-Fly, Cordylobia
anthropophaga Griinberg, in Sierra
Leone
Trichonema tetracanthum, Mehlis, 1831,
of Looss, 1900
TI. congolense transmitted by Glossina

DALAL eros ss occn eet meeot AN Sheth aie 5 «6

aby,

359
166

Cee eeeseeseseseeeeeeeseses

543

Pose eeereoseeseoeseoenes

543

COSC S HHH TE HEHEHE HOHE HEHEHE HESS EOE ED

CCHS SHOE HE HOHE EEO EEE OHH EEE

PAGE
Trypanosomiasis in the Eket District,
Southern Nigeria, April and May, 1922 93
Tumbu-Fly, Cordylobia anthropophaga

Griinbergt, «su jscpeto.d..-osee<) seer beete od 443
Typhus Fever in Greek Refugees......... 347
Uriné in;Malaria).. .25..-kizsdete sna.) 556
Venezuela, Notes on Culicidae in ...... IOI
W atsonius ASERUS 5 axhGviGal). > essai 159
W eellmauius, Genus a, Servet eee ee eran 189

Wenyon, C. M. Coccidiosis of Cats and
Dogs and the Status of the Isospora
of Man

PAGE —

White, R.O. Yellow Fever in the Gold
Coast: its Endemic and Epidemic
Character. ..9y)a4iecngsnstaen onteaestnenn 4

Wright, W. Rees. A Preliminary Account
of the Results of Surveys for Breeding
places of Mosquitoes in North Wales

Wyeomia (Decamia) pseudo-pecten, male
specimen from Venezuela...............+

Yatren, Treatment of amoebic Dysentery
PVICD, | 0020+; -maaeomeneteeesere sean

Yellow Fever in the Gold Coast: Its
Endemic and Epidemic Character ...

INDEX OF GENERA, SPECIES AND VARIETIES NEW TO SCIENCE

PAGE
Anopheles (Anopheles) amaZontcus ......4.. 72
Bothriddumcr Gauge teneccoreay tes sae: arg
CArMyertus CAOPOTIS | ea seas phan cee seo nenatet 183
Culex annultoris var. CONgOLENSIS .10..0.e0000 gI
Culex MAT ACaVensts Benes cassten tM une eee 102
CiuleX MOULD Ete aceon per pet eee 89
Culex: Pagans Vince vns cece tenes are ee 104
Davainer (Sil.) Casgatiia ee eer 47
Davainea (5.1) 1nfrequens o.cccssec se foes 55
Dilepis horvdthi went eee ee 67
Dilepis leproppallus vara cases aap eesaee 63

vili

PAGE —
Dilepis yorktt OA eI Neda ence eee 59
FAMeria: canis Whapesteccetceden ten eee 261
Forctpomyta (Euforcipomyia) papilion- q

LOOT FR a Ee 23

Tsospora' felis’ 33) Rol ee oe 248
Monopylidium freldingt ......ccccecscesceens 318
Paramonvezia Gen: }. 4. ee 329
Paramoniezia Sais PEt as rete 32908
Phlebotomus SQUAMITOSTIS .iccscccsscenecens 531m
T axorchis Gen... .csicy in tee ae 166 —

i
A

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539

| nefits BM Mace eat Se
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Publications of the
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. XVII. No. 1. April 18, 1923

A Case of Creeping Eruption in a European in the Gold Coast. By J. F. Corson. One
Plate,

Depth, and the Larvae and Pupae of Stegomyia fasciata, F. By J. W.S. Macrir, D.Sc., M.B.
A Note on the Action of Lithium Chloride on Mosquito Larvae. By J. W.S. Macrir, D.Sc., M.B
Malaria in Chimpanzees in Sierra Leone. By S. ApiER, M.B., Ch.B. Two Plates.

New and Old Observations on Ceratopogonine Midges Attacking other Insects. By
FF. W. EDWARDS.

On some Strongylid Larvae in the Horse, especially those of Cylicostomum. By J. E. W.
IBLE a oc., and. (,ieeVAN: OORDT, (D.Sc.

Avian Cestodes from New Guinea. II. Cestodes from Casuariformes. By Dr, ALEXANDER KoTLAN.
Avian Cestodes from New Guinea. III. Cestodes from Galliformes. By Dr. ALEXANDER KOTLAN,

An Anopheles of the Myzorhynchus Group (Avxopheles amazonicus, sp.n.) from South America.
By Lieut.-Col. S. R. CHRISTOPHERS, C.I.E., IL.M.S. One Plate.

The Etiology of Blackwater Fever. By B. BLackiock, M.D., D.P.H.

A New Species and a New Variety of Culex from the Belgian Congo. By A. M. Evans, M.Sc.
Report on Sleeping Sickness in Eket District, Southern Nigeria. By R. W. ORPEN.

Notes on Culicidae in Venezuela. Part III. By A. M. Evans, M.Sc,

ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY
Vol. XVII. No. 2. July 12, 1923

A Revision of the Amphistomata of Mammals. By P. A. MApPLEsTONE, D.S.O., M.B., Ch.B.
Four Plates.

Malaria in Australia. By P. A. Mapriresrone, D.S.O., M.B., Ch.B.

Coccidiosis of Cats and Dogs and the Status of the Isospora of Man. By C. M. WENyon,
C.M.G., ‘C.B.E.,) M:B.,. B.Se> Six Plates:

A Further Note on the Occurrence of Ancylostomes Resembling Necator americanus
amongst Domestic Pigs in Amazonas. By R. M. Gorpon, M.D.

A Case of Acute Ascending Paralysis in a Chimpanzee. By S, ADLER, M.B., Ch.B., and
E. J. CLARK, M.B.

ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY

Vol. XVII. No. 3. October 13, 1923.
Studies in the Treatment of Malaria—XXXI]. Summary of Studies I—XXXI. By J. W. W.
STEPHENS, M.D., F.R.S.

Notes on Australian Cestodes. By P. A. MAPLESTONE, D.S.O., M.B., Ch.B. and T. SouUTHWELLI,
M.Sc, AARC Acs Fass.

A Note on Ophiotaenia punica (Cholodovski, 1908), La Rue, 1911. By T. SOUTHWELL,
M.Sc., A.R.C.Sc., F:Z,S..and-S., ADLER; M°B;, -Gh:B:

A Pyrrhocorid Bug Capable of Biting Man. By B. BLackLock, M.D., D.P.H.

Typhus Fever in Greek Refugees. By Major-General Sir PATRICK HEHIR, K.C.I.E., C.B.,
C.M.G.7 DP Hep LM, Biv:

The Value of the Sachs-Georgi Reaction in the Serological Diagnosis of Syphilis. By
GEORGE STUART, M.A., M.B., Ch.B.

Note on Aédinus amazonensis, Lutz. By Miss ALWEN M. EvaANs, M.Sc.
The Treatment of Amoebic Dysentery. By R. M. Gorpon, M.D.

Relapsing Fever in the Gold Coast. By P. S. S—ELWyNn-CraRkKr, M.C., M.D., B.S., M.R.C.S.,
L.R.C.P., D.P.H., G. H. Le Fanu, M.B., C.M., D.T.M, and A. INGRAM, M.D., C.M.,,
MRICS sip aide Me

Carbon Tetrachloride in Filariasis. By S. ADLER, M.B., Ch.B.
Yellow Fever in the Gold Coast: Its Endemic and Epidemic Character. By R. O. WHITE.

Miscellanea: Kurloff Bodies in Fish—Trichonema tetracanthum, Mehiis, 1831, of Looss, 1g900—Pigs
and Ankylostomiasts in the Gold Coast—Onchocerca armillata in Cattle in the Gold Coast.

ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY
Vol. XVII. No. 4. December 21, 1923.
A Study of the Tumbu-Fly, Cordylobia anthropophaga, Grinberg, in Sierra Leone. By
B. BLACKLOCK, M.D., D.P.H., and M. G. THoMpPSON. Four Plates.

The Transmission of T. congolense by Glossina palpalis. By H. LyNpHurst DvKE, O.B.E.,
MD, 3c.D2 Cantap.

On a New Species of Phlebotomus from Japan. By R. NewstTrap, F.R.S.

Glossina ziemanni, Grinberg, a Synonym of Glossina palpalis, Sub-species Fuscipes,
Newstead. By R. NEWSTEAD, F.R.S.

Malignant Growths in Natives of Sierra Leone. By S. ADLER, M.B., Ch.B., and E, H. TAYLOR
CUMMINGS.

A Preliminary Account of the Results of Surveys for Breeding-places of Mosquitoes in North
Wales. By W. REES Wriaut, B.Sc.

The Frequency of Indicanuria. By R. M. Corpo, M.D.
}

Miscellanea: Notes on Cestode Parasites from a Duck—Notes on Parasitic Worms from the Golé
Coast—Cittotaenta largorchestis, Lewis, 1914—Davainea leptotrachela, WHung., 1910—
Param phistomum cervt in a horse—A Note on Auchmeromyia luteola, Fab.—Ancylostomum
cevlanicum in Cats and Dogs of South India—The Urine in Malaria.

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Volume XVII December 21, 1923 No. 4

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.
Proressor Re NEWSTEAD S@ MSc.,4):P., F:R.S., ACLS; 2.ES.,) Honssh;R.H.5.

Proressorn WARRINGTON YORKE, M.D.
Prorressor B. BLACKLOCK, M.D.

THE INCORPORATED
LIVERPOOL SCHOOL OF TROPICAL Shi sia

FoUNDED BY SIR ALFRED LEWIS JONES, K.C.M.G.
(Affiliated with the University of Liverpool)

Hon. President: H.R.H. THe DUKE oF York, K.G., G.C.V.O.
Chairman: SiR FRANcIs C. DANSON
Vice-Chairman: Professor R. CATON, C.B.E., M.D., F.R.C.P., LL.D.

Hon. Vice-Presidenits: Tur Eart or Dersy, K.G., G.C.V.O., C.B., LL.D.
VISCOUNT MILNER, G.C.B.
VISCOUNT we PRRIE, pike lo eer,
VISCOUNT LEVERHULME
BARON KYLsAntT, G.C.M.G.
SIR EDWARD MEREWETHER, K.C.V.O.
Mr. O. HARRISON WILLIAMS

COMMITTEE

Sir Hee] READ AW CG: Colonial Office

Vice-Chancellor J. G. ApAmt, C.B.E., — :
Msp Rika:

Professor R. Caton, C.B.E., M.D.,
PRG Ber we)

Mr. H. WADE DEAcoN, C.B.E.

University of Liverpool
|
)
Professor J. M. BEaTTIE, M.A., M.D.,
|

Council of University of Liverpool

Mu ALT Gee See
Professor W.. J /.DAKIN,} D.Sc., FXILSS
Boz
Mr. E. G. BUCKLEY
Mr. T. WooDSEND

Senate of University of Liverpool

Royal Southern Hospital

Mr. ENFIELD E. FLETCHER Steamship Owners’ Association
Mr. W. J. B. CHAMBERS ) ; soe
Mir Galenene j Shipowners’ Association

Mr. G. BROCKLEHURST

Mr. H. D. DICKIE

Professor. E.3E. GuyYnn, MAl -M:D. C MARR Pe eG.
Professor Sir W. HERDMAN, C.B.E., D.Seo FoR Se Pel ss., bR‘SE:
Professor E. W. Hore, O.B.E., D.Sc., M.D.

Mr. Davip JONES

Mr. J. PICKERING JONES

Captain R. RANKIN

Mr. O. HARRISON WILLIAMS

Professor J. W. W. STEPHENS, B.A., M.D., F.R.S.

Professor R. NEWSTEAD, M.Sc. 2 ROS. ALL So EES) Hon. ER Hes)
Professor W. YorKE, M.D.

Professor B. BLacKLock, M.D., D.P.H., D.T.M.
Mr. J. A. TINNE, Hon. Treasurer

Mr. J. L. McCartuy, Secretary J. MippDLEMASS HUNT,
H 24—25, Exchange Buildings, Liverpool Hon. Dean

Alfred Jones Professor of
Tropical Medicine .

Dution Memorial io °f
Entomology

Walter Myers aia of
Parasitology

Staff, 1923

JOHN WILLIAM WATSON STEPHENS, B.A., M.D., Cantab.,
F.R.S.

BOBLE Pe NEWS FREAD. I.P.oF.R.S:, MiSc, “A.L-S.; F.E,S.,
‘ Hon. F.R.H.S.

. WARRINGTON YORKE, M.D.

Professor of aia Diseases of

Africa .

Lecturer on Entomology

. BREADALBANE BLACKLOCK, M.D., D.P.H., D.T.M.
. ALWEN M. EVANS, M.Sc.

Assistant Lecturer on Entomology . W. H. POTTS, B.A.

Lecturer on Protoxoology
Lecturer on Helminthology
Clinical Pathologist

Hon. Lecturer on Clinical
Veterinary Parasitology

SlaWeccor | VAC rib IvMen ws Wisb.>, Cheb... oc.
ot nO UTRWEL I M.Sc:,JA-R:C.Sc.,F.Z.5.
. RUPERT MONTGOMERY GORDON, M.D., Ch.B.

- A. W. NOES "PILLERS, F.R.C.Y-.S.

Assistant Lecturer and Demonstrator

on Veterinary Parasitology

Lecturer on Tropical Surgery

au acalit.

» ROBERT ERNEST KELLY, C:BseMsD. BSc, kcc's:

Lecturer on Tropical Sanitation . R. H. KENNAN, B.A., M.D., F.R.C.S., D.P.H., D.T.M.
Hon. Lecturer on Tropical Sanitatiox WILLIAM THOMAS PROUT, M.B., C.M.G., C.M.

Hon. Statistician

Physician .

Assistant Physician

Consulting Surgeon

Director . i

Director . 4
Assistant Director

Research Assistant

4 WALTER: SLOTT.

Royal Infirmary, Liverpool

. JOHN WILLIAM WATSON STEPHENS, B.A., M.D., Cantab.,
E.RS.

. WARRINGTON YORKE, M.D.
. W. THELWALL THOMAS, Ch.M., F.R:C.S.

The Manaos Research Laboratory

. HAROLD WOLFERSTAN THOMAS, M.D., C.M.

Sierra Leone Research Laboratory

. BREADALBANE BLACKLOCK, M.D., D.P.H., D.T.M.
. PHILIP A. MAPLESTONE, D.S.O., M.B., Ch.B.
. S. ADLER, M.B., Ch.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. Haftkine
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

19t3s—

Professor Fred V. ‘Theobald

ba by fo

Dr. Griffith Evans

19 19—

Dr. J. W. Scott Macfie
The Oswaldo Cruz Institute, Rio de

Janeiro

1920—

Major E. E. Austen, D.S.O.

Dr, A. G. Bagshawe, C.M.G.

Dr. Andrew Balfour, C.B.

DroA. LG. 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 Farmer Allen
1923—
John Cecil Cruickshank

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... = a te Twenty Guineas
Short, Advanced Course ... Aah ee Six Guineas
Course in Veterinary Parasitology no Fifteen Guineas
Diploma Examination a. ie 1 Five Guineas
Fee for the use of a School microscope during 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 Maddock, Edward Cecil Gordon

1904 Bennett, Arthur King 1905 Moore, James Jackson

1904 Bruce, William James 1905 Nightingale, Samuel Shore

1904 Byrne, John Scott 1905 Radcliffe, Percy Alexander Hurst

1904 Clayton, Thomas Morrison 19e5 Young, John Cameron

1904 Dalziel, John McEwen

1904 Dee, Peter 1906 Adie, Joseph Rosamond

1904 Greenidge, Oliver Campbell 1906 Arnold, Frank Arthur

1904 Hehir, Patrick 1906 Bate, John Brabant

1904 Khan, Saiduzzafor 1906 Bennetts, Harold Graves

1904 Laurie, Robert 1906 Carter, Robert Markham

1904 Maclurkin, Alfred Robert 1906 Chisholm, James Alexander

1904 McConnell, Robert Ernest 1906 Clements, Robert William

1904 Nicholson, James Edward 1906 Dundas, James

1904 Philipson, Nicholas 1906 Faichnie, Norman

1904 Sharman, Eric Harding 1906 Jeffreys, Herbert Castelman

1904 Thomson, Frank Wyville 1906 Mackenzie, Donald Francis

1904 Walker, George Francis Clegg 1906 Pailthorpe, Mary Elizabeth
1906 Palmer, Harold Thornbury

1905 Anderson, Catherine Elmslie 1906 Pearse, Albert

1905 Brown, Alexander 1906 Sampey, Alexander William |

1905 Caldwell, Thomas Cathcart 1906 Smithson, Arthur Ernest

1g05_ Critien, Attilio 1906 Taylor, Joseph van Someron

1905 Hooton, Alfred 1906 Taylor, William Irwin

1905 Hudson, Charles Tilson 1906 Tynan, Edward Joseph

1905 Illington, Edmund Moritz 1906 Watson, Cecil Francis

1905 Macfarlane, Robert Maxwell 1906 Willcocks, Roger Durant

Date of
Diploma
1906

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1908
1908
1908
1908

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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

Samuel, Mysore Gnananandaraju

Shroff, Kawasjee Byramjee

Thornely, Michael Harris

Date of
Diploma
1909
1909
19°99

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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 Balabha
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
Ievers, 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 Ahobaia

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
Milne, Arthur James

Mitra, Manmatha Nath
Myles, Charles Duncan
Pelly, Huntly Nevins

Date of
Ditloma

1gI2
1912
1gI2
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1913
1913
1913
1913
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1916
1916
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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 Francis

Yin, Ulg Ba’

Young, William Alexander

Arculli, Hassan el
Chohan, Noormahomed Kasembha
Connell, Harry Bertram
Gerrard, Herbert Shaw
Gimi, Hirji Dorabji
Gwynne, Joseph Robert
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

Barseghian, Mesroob
Chaliha, Lakshmi Prasad
Lim, Albert Liat Juay
Lim, Harold Liat Hin

Date of
Diploma
1916
1916
1916

1917
1917
1917

1918

Peg
Bg ko
1929
1919
eB NS
rgt9
r919
S049
1919
gto
1gt9
1929
|
eo
tO1G
LOTS

1920
1920
1920
1920
1920
1920
1920
1920
1920
1920
1920
1920
1920
1920

1921
1921
1921
1921
1921
1921
1921
1921
1921
1921
1921
1921
1921

1922
1922
1922
1922
1922
1922
1922
1922
1922
1922
1922
1922

1923
1923
1923
1923
1923
1923

Metzger, George Nathaniel
Séderstrém, 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 Frederick
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

Basu, Dhirendranath
Cruickshank, John Cecil
Doherty, Winifred Irene
Elsohn, John

Raja, Rojaporum
White, Charles Francis

ANNALS OF TROPICAL MEDICINE
AND PARASITOLOGY

EDITORIAL NOTICE

Articles tor 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.—Blue -
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 sujficzently 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.

SMITH, 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.

CONTENTS hee

| Rae PAGE
Bracktocx, B, and Tagen M. Gy A Study ok the TabnbacFly,

Cordylobia anthropophaga Grinberg, i in Sierra Leane hy). tee) 44307

By Doe, ‘H. Lynpuursr. The Transmission of T. congolense. re Hla

_ palpalis ity aa oa pas na eee Shape. , o¥e a2 @ j iy std bas . hy Sib

oa “EDITORIAL. NOTICE

nice: Ciee! publication should not. exceed. 25 pages ot the

es and will be understood to. be offered alone to this Jarier.
a ney: should pe typewritten and addressed tole

ee ae |The, Editors

Renee! of Tropical Medicine
aie. ae c Pore e Place:
Le ae oe s Liverpool

Paaeaid: ee text Creed: Or ee should be drawn eledile and

| “ ey in Indian ink, if possible, on Bristol board. N.B:—Biue or other.

We coloured ruling in squares or lines cannot be reproduced,

enue AIP lettering, names or legends on text-figures, charts or maps should

Ba be printed: sufficiently large to allow of clear degibility of reduction

ke. us peteett

Plates and sfteatra mune should 1 accompanied by. short explanations,

References to authors in the text must be made in the fol lowing way :>—

| a hanine to. Smith (1900) the ‘spleen. is enlarged, but Robinson (igi4)

_ says the reverse,’ The references should be collected in alphabetical.

| Newsrzap, R.. ‘On a New Species bs Pblebotomas from bo ee y ie Sap ae,
if ‘Newsrzap, R. Ney the: Ziemannt. Griinberg, a Srtenym of. Glassina Rie
“palpalis, Sub-species P wscipes, SVEWSECAG A Mo caf ames fea tliin vex oy OSs
aN fe Die S., and Cummines, E, Hy Tavton. | Malignant Suits in Natives Shee:
BON a) Sierra Leone: h ve Lied aD Reape tas ete Mamet SEM Me sg nage aR
| Wrens, W. Res, uf Peck dint acinar fe ai Roots of Surveys £ for we
v, see acd of Mosquitoes i in North Wales. RA ea Ror ateen ORION
uy Goxnon, te |The e, Ereqnency of of Indicanoria we re eo eae 2 549)
gene Lae ee anne wee Be eae oe Apes heey orden ee

order of authors’. ‘surnames at the end. ae Be, paper, and arranged in the | «

ch 3 following way: nae Sure ape Sd a a

ihe es eh -Rosinson, 3: dla}: scTha sateen in malin’ Annals or aslogy,

Vol. XX, pp. 20-25. .
Suita, J- (1900). » ‘Enlargement of hae spleen in ‘malatia,” - Fournal oy
_. Pathometry, Vol. ay. pp- 1-20. |

wenty-iive reprints are supplied of each paper, free of charge.
R Additional neon (ep to a can Be Seana at cost price.’ "

Subeehanea’ oe 2s. yee per See neee free, bavie in
Padeasice: to the Secretary, The University Press of Liverpool

} Limited, 177 Brownlow Hill, Liverpool, to whom obs Feopondpace |
as cane tae Pay erereracnts. should aieg. be Ps nabincits '

nee Tuan & & Co. re PRINTERS, 53 Vicronra Sonar, I. IVERPOOL.

ESREEEE

a
ef Fe.

ee
7

gray

er Bureau

Nie ce 1, an and Le

Ey appl id OF VEL ow 'REVER
EXPEDIT ION TO. YUCATAN, ate

may be purchased at 6s each.

Haar

Yelow Fever B

‘ oe th YELLOW eee COM
MISSION ON INVESTIGATION OF NON
MALARIAL eee iN WEST AFRICA,

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UNIVERSITY OF ILLINOIS-URBANA

616. 9605L! C001
ANNALS OF TROPICAL MEDICINE AND PARASITO
17 1923

301120

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