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Journal of Parasitology 


A QUARTERLY DEVOTED TO MEDICAL ZOOLOGY 


EDITORIAL BOARD 


FRANKLIN D. BARKER ALLEN J. SMITH 

The University of Nebraska The University of Pennsylvania 
CHARLES F. CRAIG JOHN W. SCOTT 

Medical Corps, U. S. Army The University of Wyoming 
WILLIAM B. HERMS CHARLES W. STILES 

The University of California U. S. Public Health Service 
BRAYTON H. RANSOM RICHARD P. STRONG 

U. S. Bureau of Animal Industry Harvard Unicersity 
WILLIAM A. RILEY JOHN L. TODD 

Cornell University McGill Unicersitp 


ROBERT T. YOUNG 
The University of North Dakota 


VOLUME 2 & 


1916-11A ¢ 
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Managing Editor 
HENRY B. WARD 


The University of Illinois 
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CONTENTS OF VOLUME II 


SEPTEMBER, 1915. NUMBER 1 
PAGE 


ON THE OCCURRENCE OF A TRYPANOPLASM, PRoBABLY Trypanoplasma borreli 
LAVERAN ET MESNIL, IN THE BLoop oF THE CoMMON SUCKER, Catostomus 
ene ReaEAONE SRT Oe ee c's au eels sco ce nese sevcese 1 

(With one plate) 


Two New Cases or PoLyrRaADIATE CESTODES, WITH A SUMMARY OF THE CASES 


ATMEADN Owns VV INTHROP 1); “ROSTER. 2.0... .. ccc essen ccesscecans 7 
Some Notes aNp EXPERIMENTS ON Sarcocystis tenella RAtLuiet. JoHn W. 

215 1) ea Sees SAY a BR eer eT Sesion es Pec aismte oensle 20 
Ecc VARIATION IN A TREMATODE SPECIES. WILLIAM WALTER Cort........ 25 


SomE New GREGARIMXE PARASITES FROM ARTHROPODA. MINNIE ELIZABETH 
TIES ee ee 6 al) PES S88 ae Se aan pene a 27 
(With two plates) 
Pneumonyssus foxi Nov. sp., AN ARAGHNOID ParRASITIC IN THE LUNG OF A 


Monkey (Macacus rhesus). Frep D. WEIDMAN...............0000005 37 
(With one plate) 


Grestonpe Cysts From MuUsKRAT. EDWIN LINTON.......-...020+sceceessne+s 46 
SARCOPHAGID LARVAE FROM THE PAINTED TurRTLE. F. E. CHIDESTER....... 48 
PVGTIOS 1.2 wedtt Se deem Nea Soin hn eee PE ic tae ete en 50 


DECEMBER, 1915. NUMBER 2 


RaeOOEa IS SS (OURO NER AVE BK ed Won seay dget ava ebsfevovekayhay ate cictovars ototatereversterdretetelatete oe woke oe 51 
(With portrait) 
FurTHER Note Upon Comparison or Endamoeba gingivalis (Gros) AND 
Endamoeba histolytica SCHAUDINN. ALLEN J. SMiTH AND M. T. Barrett 54 
NOTES ON THE TREMATODE GENUS TELORCHIS WITH DESCRIPTIONS oF NEW 


Sa MEMES PACT EI ANY (TRIM ROAREN 52 os as Ga es ddd dee Se ace wnwaee eis 57 
(With one plate) 


THe Insect Vector or Uta, a PeruviAN Disease. CuHartes H. T. 


WOWNSEMD) Jc Weak vid. seks ne es SPE eee EE Nach sieMk Wisi siexclarhed wep aces 67 
Filaria cingula Parasitic IN THE SKIN oF Cryptobranchus alleghéniensis. 
[Fir tpg Safer US Fe 74 
THE Lire History or Gongylonema scutatum. Brayton H. RANSOM AND 
ease OE AD Ea eine ee alk ols wes ida ci viale sis ce ewan ows 80 
Note oN THE Stace or Piroplasma bigeminum WuicHh Occurs IN THE 
CattLteE Tick, Margaropus annulatus. Howarp CRAWLEY........-....- 87 
THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON, PROCEEDINGS..........--- 93 


DHGET aS iret ath Bt ore, Yo cre ty a Se's.-dvapShaneey hetetorete Ln odes ool b SR eee eee 96 


MARCH, 1916. NUMBER 3 


PAGE 
TA FAMILLE DES PRELAZIIDAE.. A. RATELIET... 5.0.6 site 2. oop 99 
SEASONAL DISTRIBUTION OF SOME ACANTHOCEPHALA FROM FRESH-WATER 
Hosts. H. J. VAN CLEAVE.........0. 22-0 e eens cee cs ences essneeneseres 106 
On THE INTERMEDIATE Hosts or THE LuNG Distome, P. westermanii 
IOKREERT © ADAG: VOSEUIDA .. 2 sicysjc sc 0.0.00 srapeip Rta tenie\ ohelslc stetn orate eel nee 111 


(With one plate) 


GONGYLONEMA IN THE ROLE or A HuMAN Parasite. Henry B. Warp.... 119 
(With one plate) 


Are SARCOSPORIDIA ABERRANT ForMS oF CNIDOSPORIDIA OF INVERTEBRATES? 
BG ATET= VATERIO® oc. co. < sce css! 6-05! he Ree ee SP ae ts 126 


THREE New GREGARINES FROM MARINE CrusTACEA. MINNIE E. Watson.. 129 
(With one plate) 


Tue Payaroetto Tick (Ornithodorus coriaceus KocH) WITH SPECIAL REF- 


ERENCE TO LirE History AND Bitrnc Hasirs. WutitAMm B. Herms.... 137 
Note ON THE ETIOLOGY oF VERRUGA AS DEDUCED FROM A STUDY OF THE 

ASEXUAL STAGES OF BARTONELLA. CHARLES H. T. TOwWNSEND........ 143 
A New InrFusorIAN ParASITE IN SAND FLEAS. MINNIE E. WaATSON...... 145 
REVIEWS oc biversrite-d eric ckise ele owls os asa. 5 oud b5 0 eee Sis oie eh bee ele ee eee ee 147 


JUNE, 1916. NUMBER 4 


THE SIGNIFICANCE OF CERTAIN NATURAL FLAGELLATES OF INSECTS IN THE 
EvoLuTIoN oF DISEASE IN VERTEBRATES. H. B. FANTHAM AND ANNIE 
PORTER mck Je Gant cdioaa ee or 


A REVISION oF THE GENUS ARHYTHMORHYNCHUS. H. J. VAN CLEAVE...... 167 
(With two plates) 


Some Notes oN THE EnNcysteD LARVA OF THE LUNG DistToME. SADAO 


YOSHIDA. ef oso ecb. oi bie < ocd ae nts SIO Oe oe eee 175 
Cylindrotaenia americana Nov. SPEC. FROM THE CRICKET Froc. Munna. E, 
JE WELL «oo: oa osa:e nig ore oe ib ce 0 ve wm pie SETI OR OE EE ese seo ea 181 
(With one plate) . 
THE-Errect oF Tick. Bires on MAN. BD; McCArFREY. <3»: o--14)- 6-2 eee 193 
Tue HELMINTHOLOGICAL SOCIETY OF WASHINGTON, PROCEEDINGS...........--- 195 
REVIEWS. AND NOTES J o:0600c0 00 26 Bere ee oe Be he cle eis: ae ae 201 


: vaio numbers of Volume II of the JouRNAL of PARASITOLOGY were mailed as 
ollows : 


No. 1. Nov. 16, 1915 No. 3. May 9, 1916. 
No. 2. Jan. 29, 1916. No. 4. Aug. 2, 1916. 


EXPLANATION OF PLATE 


Trypanoplasma, probably 7. borreli. All the figures with the exception of 
Figure 1, which is a free-hand drawing, were drawn with the camera lucida, 
using a Leitz 2 mm. apochromatic objective and compensating ocular X 18. 
They have been reproduced at a magnification of 2,600 diameters. Figures 2-6 
were drawn from smears fixed with osmic acid vapor, stained with Giemsa’s 
azur-eosin and mounted in neutral Canada balsam. 

Fig. 1. Drawn from a living specimen in a fresh preparation of the blood. 
Fig. 2. From blood from the kidney. 

Fig. 3. From blood from the heart. 
Figs. 4 to 6. From blood from the kidney. 


The Journal of Parasitology 


Volume 2 SEPTEMBER, 1915 Number 1 


ON THE OCCURRENCE*OF A TRYPANOPLASM, PROB. 
ABLY TRYPANOPLASMA BORRELI LAVERAN ET 
MESNIL, IN THE BLOOD OF THE COMMON 
SUCKER, CATOSTOMUS COMMERSONIP 


J. W. Mavor 


University of Wisconsin 


In spite of the considerable interest in the distribution of the hemo- 
flagellates, there is no record, so far as the writer is aware, of a 
trypanoplasm occurring in the New World. The species to be described 
seems to be identical with Trypanoplasma borreli described by Laveran 
and Mesnil (1902). This species is reported to cause disease and 
death in fish in captivity, but no case is recorded of a trypanoplasm 
causing a pathogenic condition in a fish in the wild state, where there 
is little opportunity for more than a single infection. On this account 
the facts to be recorded are of special interest. 

The sucker in which the trypanoplasm was found was seen in 
shallow water at a wharf in Go Home Bay, a small bay leading from 
the Georgian Bay, about twenty miles from Penetanguishene, Ontario. 
The fish was sluggish and allowed itself to be easily picked up in a 
dip net. When brought to the laboratory and taken out of the water 
it died in a few minutes. There were no external lesions and no 
abnormalities were discovered in a hasty examination of the viscera. 
The gills were pale and bloodless. Preparations of blood from the 
heart showed the presence in great abundance, one or two in a single 
field of the immersion objective, of the trypanoplasm to be described. 

As the Biological Station was about to be closed for the season it 
was possible to obtain only one other sucker, which was caught in a 
fyke net. This was active and normal in every way. In five fresh 
preparations of the blood from the gills, the heart, and the liver no 
hemoflagellates were found after a careful search. 

The evidence available in this case is scarcely sufficient to prove 
the trypanoplasm as a specific cause of the pathogenic condition of the 


1. The observations in this paper were made while the writer was curator 
of the Biological Station of the Canadian government at Go Home Bay, near 
Penetanguishene, Ontario, in 1913. The courtesy of the directors of the Bio- 
logical Board of Canada permits their publication here. 


2 THE JOURNAL OF PARASITOLOGY 


fish. The parasite is interesting, however, in that it occurs in a fish 
in the wild state, where there is little probability of there being more 
than a single, or at most, a few infections, the parasite being without 
doubt carried by a leech. There is no record, so far as the writer is 
aware, of a trypanoplasm causing a pathogenic condition in a fish in 
the wild state. Such a condition, however, has been recorded for fish 
in captivity, and if, as the author is inclined to believe, the parasite of 
the sucker is T. borreli, for the same species of parasite. In carp, 
Cyprinus carpio L., infected with T. borreli, Marianne Plehn 
(1904:175) finds: “Bei stark befallenen Fischen erreicht die Anamie 
einen ganz extremen Grad; man kann nur wenige Tropfen eines was- 
serigen, kaum rotlichen Blutes gewinnen; Keimen und innere Organe 
sind ausserst blass. Andere pathologisch-anatomische Merkmale 
fehlen. Die Tiere zeigen in der letzten Lebenszeit ausser beschleunig- 
ter Atmung und grosser Unlust sich zu bewegen nichts Auffalliges. 
Sie gehen offenbar an Blutmangel ein, den sie, zwar lange, aber doch 
nicht dauernd ertragen koénnen. Es ist unzweifelhaft, dass die Krank- 
heit auch im Freien Schaden anrichten wird; die Beobachtungen sind 
noch zu jungen Datums um allgemeine Angaben uber Verbreitung und 
Bedeutung zu gestatten.” Keysselitz (1906) has also described the 
pathological condition in the carp due to T. borreli. Leger (1904 :824) 
describes cases of acute infection of the minnow, Phoxinus laevis 
Agass, with a trypanoplasm as follows: “Des infections aussi intenses 
aménent chez le poisson une anemie profonde: decoloré et enflé il se 
tient immobile, refuse toute nourriture et finit par mourir.” A case 
in which a trypanosome may cause a pathogenic condition has been 
recorded by Doflein. He remarks (1909:398), referring to Trypano- 
soma carassii: “Ich selbst hatte allerdings einmal Gelegenheit, eine sehr 
ahnliche, vielleicht sogar identische Art im Blut der Schleihe, Tinca 
vulgaris, zu beobachten ; die befallenen Schleien waren offenbar krank, 
sie waren sehr apathisch und waren an die Station zur Untersuchung 
von Fishkrankheiten in folge eines grossen Sterbens in den betreffen- 
den Weihern eingesandt worden.” 

The terminology used in this paper is that used by Minchin (1912), 
with the exception of the term “basal granule,’ which has been used 
in place of the term “blepharoplast,” as used by that author. 

As seen in the living state (Fig. 1), the trypanoplasm has the form 
typical of the genus; a thick yielding body with two flagella, one of 
which forms the border of an undulating membrane. The measure- 
ments of the body are, length 20-25 p, thickness 3-4 ». The anterior 
flagellum is between one-half and two-thirds as long as the body. The 
posterior flagellum arises near the anterior flagellum and passes pos- 
teriorly, forming the margin of the undulating membrane. It extends 


MAVOR—TRYPANOPLASM IN BLOOD OF SUCKER 3 


freely for about two-fifths of its length beyond the posterior end of 
the body. The undulating membrane is comparatively thick and not 
sharply distinguishable from the edge of the body. The parasites 
showed an active writhing motion, but little progression. What there 
was seemed to be with the morphologically posterior end in advance. 
The protoplasm was finely granular, a few larger highly refractive 
greenish granules measuring up to 0.4 » in their longest diameter, being 
present in the posterior half (Fig. 1). After the preparation had been 
standing for a little time, sealed with vaselin, two or three large 
vacuoles were seen in the posterior end of some individuals. 

Smears of the blood from the heart and from the kidney were 
fixed in the vapor of osmic acid, stained with Giemsa’s azur-eosin and 
mounted in Canada balsam neutralized with lithium carbonate. In 
each of the smears the trypanoplasm was found to be abundant. 
Although often much distorted, the parasite is found in some parts of 
the smears remarkably well preserved. It is to be regretted that 
when the parasite was discovered time did not permit of the making of 
“wet” smears stained with hematoxylin. The results especially as 
regards the kinetonucleus are open to criticism on that account. 

About fifty of the best-preserved and most clearly showing indi- 
viduals have been studied in detail with a 2 mm. apochromatic objec- 
tive and compensating oculars 12 and 18. The parasites show great 
uniformity in size and general structure. They are nearly always 
sickle-shaped, the trophonucleus and the undulating membrane being 
on the convex side. Measurements show little or no difference in size 
between fresh and preserved individuals. 

The protoplasm, as in the fresh preparations, is finely granular and 
contains a varying number of relatively large, deeply staining granules 
distributed either mainly in the posterior end (Fig. 3), or irregularly 
throughout its extent (Figs. 4, 5). It is possible that these larger 
granules are identical with the greenish granules observed in fresh 
preparations. Such “chromatoid granules” have been found in dif- 
ferent species of trypanoplasms (Leger, 1904; Keysselitz, 1906; 
Friedrich, 1909; Minchin, 1909). It is doubtful whether these granules 
are chromidial in nature. 

The anterior flagellum arises at the extreme anterior end and on 
the side of the body on which the kinetonucleus is located. It leaves 
the body independently of the posterior flagellum (Figs. 4,5). The 
posterior flagellum arises very close to the anterior and passes around 
the blunt anterior end and along the entire length of the body as the 
margin of the undulating membrane (Figs. 1, 2). It is continued 
posteriorly as a free flagellum of a length equal to about two-thirds 
that of the body. The undulating membrane shows in some cases as 


a THE JOURNAL OF PARASITOLOGY 


a clear unstained area between the posterior flagellum and the granular 
protoplasm of the body (Figs. 2, 4). 

The kinetonucleus (Woodcock, 1909, for the “Geisselkern” of 
German authors) is situated on the side opposite to the trophonucleus 
and the undulating membrane and about one-third of the length of the 
body from the anterior end. It has a clear outline and stains deep 
purple, in contrast to the more reddish tinge of the trophonucleus. In 
the individuals which show least distortion it is ovoid, about half again 
as long as it is wide and shows a distinct membrane. Its size varies 
between wide limits (Figs. 2-5). Its usual size, however, is 3.5 by 
2.4 ». When not too deeply stained five or six deeply staining bodies 
can be seen lying immediately under the membrane. In some cases 
what appears to be two kinetonuclei are present in individuals which 
show no division of the flagella or trophonucleus (Figs. 3, 5). In such 
cases each of the two bodies shows a clear contour and is undoubtedly 
surrounded by a membrane. Each also shows the included stained 
granules, as in the case of the single kinetonucleus, but the number 
of granules in each is less than in the single kinetonucleus. The two 
bodies may be of almost equal size, or one, always the anterior, may 
be much the smaller (Figs. 3, 5); they may be near together or far 
apart. 

That this dual nature of the kinetonucleus is due to faulty technic 
seems hardly possible, in view of the fact that the two parts are 
surrounded by distinct membranes. It may be that it is due to division, 
the kinetonucleus, in this case, having completely divided before either 
the flagella or trophonucleus. Against this assumption are the facts: 
first, that the individuals show no other signs of division, unless, 
which is doubtful, the presence of two basal granules is to be taken as 
such ; and second, that the two parts of the kinetonucleus are often of 
very unequal size. . 

Keysselitz (1906) finds in Trypanoplasma borreli Laveran et 
Mesnil that the “blepharoplast” (kinetonucleus) divides transversely, 
but says (p. 32): “Ein zeitlich gesetzmassiges Verhalten zwischen der 
Teilung des Kernes und Blepharoplasten kann ich nicht konstatieren.” 
Friedrich (1909:385) finds that the division of the kinetonucleus is 
“eine einfache Langsspaltung.” His figures 19 and 22 show that it 
may divide before either the trophonucleus or the flagella. Martin 
(1910) finds in Trypanoplasma congeri Elmhirst and Martin that the 
divisions of the flagella and the trophonucleus precede that of the 
kinetonucleus. 

The condition found in the blood of Catostomus commersonti seems 
to resemble most closely that described by Keysselitz (1906:25) for 
Trypanoplasma borreli in “geschwachten anamischen Fischen” (see his 
Figs. 40, 42, 451). Here, however, the kinetonucleus may be divided 


MAVOR—TRYPANOPLASM IN BLOOD OF SUCKER 5 


into more than two parts. In this connection it will be remembered 
that the sucker in which the trypanoplasm was found showed an anemic 
condition similar to that described by Keysselitz. The same author 
(1906 :37 and Fig. 47) finds in a parasite of the stomach and adjacent 
parts of the alimentary tract, “Bei Trypanoplasma ventriculi weist der 
Blepharoplast sehr haufig eine Sonderung in zwei Stiicke auf.” 
Laveran and Mesnil (1902) in their description of 7. borreli figure 
two individuals (p. 491, Figs. 13 and 15), each with two kinetonuclei. 
(Although Laveran and Mesnil considered these bodies to represent 
the trophonucleus “le noyau” and not the “centrome des Trypano- 
soma,” there is no doubt that they were mistaken. ) 

Two basal granules (centrioles, blepharoplasts of Minchin, 1912) 
are usually to be seen where the flagella arise. They stain deeply and 
are to be distinguished only by their position from the chromatoid 
granules found in other parts of the protoplasm. Although the ends 
of the flagella can in certain animals be seen to enter the protoplasm 
separately (Figs. 4, 5, 6), they cannot be traced to separate granules 
(Figs. 4, 5). 

The trophonucleus is situated about a third of the length of the 
animal from the anterior and often lies side by side with the kineto- 
nucleus (Figs. 2, 3, 5); in other cases it is slightly behind the kineto- 
nucleus (Figs. 4,6). Its shape and size resemble that of the kineto- 
nucleus, being however usually slightly smaller. It is ovoid and 
measures on the average 2 by 3 ». In many cases it shows a distinct 
membrane (Figs. 2,4). In other cases such a membrane was not to be 
seen, probably on account of poor fixation. It contains a varying num- 
ber of deeply staining granules. In some cases one of these granules 
(karyosome?) is larger and centrally located. 

So far as the writer is aware the genus Trypanoplasma has been 
described only from European fishes. The species recorded as occur- 
ring in the blood are: 
abramidis Brumpt 1906. 
barbi Brumpt 1906. 
borreli Laveran et Mesnil 1902. 
cyprint Plehn 1904. 
guernei Brumpt 1906. 
gurneyorum Minchin 1909. 
keysselitsi Minchin 1909. 
truttae Brumpt 1906. 
varium Leger 1904.4 

The species abramidis, barbi, cyprini, guernei, gurneyorum and 
truttae, as described by their authors, have a rod shaped kinetonucleus 
and the free portion of the posterior flagellum either half as long 


‘ 


le abe ete Me as ee 


6 THE JOURNAL OF PARASITOLOGY 


(barbi) or less than half as iong as the anterior flagellum ; two charac- 
ters which exclude the Trypanoplasma of the sucker. The latter 
differs, also, from T. keysselitzi which has the two nuclei near together 
at the anterior end and the kinetonucleus “very elongated”. There 
seems some doubt as to whether T. varium is not the same species as 
T. borreli, the chief argument of Leger (1904) being that the two 
forms show a preference for different hosts. 

The trypanoplasm found in the sucker has all the morphological 
characters described and figured for T. Borreli by Laveran and Mesnil 
(1902), size and shape of the body, position and shape of the nuclei, 
and length of the flagella. The writer therefore provisionally identifies 
it with this species. 

It is interesting to note that German carp, in which Keysselitz 
(1906) has studied T. borreli, have been introduced into the Canadian 
lakes and occur near where the sucker was found. Catostomus com- 
mersonit and Cyprinus carpio are closely related fish, being in the 
same family, Cyprinidae. 

It is therefore not improbable that 7. borreli has been introduced 
into the Canadian lakes with the German carp. 


PAPERS CITED 


Brumpt, E. 1906. Mode de transmission et evolution de Trypanosomes des 
poissons; description de quelques especes de Trypanoplasmas des poissons d’eau 
douce; Trypanosome d’un crapaud africian. C. R. Soc. Biol., 60: 162. 

Doflein, F. 1909. Lehrbuch der Protozoenkunde. Jena. 

Friedrich, Ludwig. 1909. Uber Bau und Naturgeschichte des Trypano- 
plasma helicis, Leidy. Arch. f. Protistenk., 14: 363. 

Keysselitz, G. 1906. Generations-und Wirtswechsel. von Trypanoplasma 
borreli Laveran et Mesnil. Arch. f. Protistenk, 7: 1. 

Lavern, A., and Mesnil, F. 1902. Des Trypanosomes des Poissons. Arch. 
f. Protistenk., 1: 175. 

Leger, Louis. 1904. Sur la morphologie du Trypanoplasma des Vairons. 
C. R. Acad. Sci. Paris, 138: 824. 

1904a. Trypanoplasma varium n. sp. C. R. Soc. Biol., 57: 345. 

Martin, C. H. 1910. Observations on Trypanoplasma congeri. Part I. 
The Division of the Active Form. Quart. Jour. Micr. Sc., 55: 485. 

Minchin, E. A. 1909. Observations on the Flagellates Parasitic in the Blood 
of Freshwater Fishes. Proc. Zool. Soc. London, p. 2. 

1912. An Introduction to the Study of the Protozoa with Special Refer- 
ence to the Parasitic Forms. London. 

ae Marianne. 1904. Trypanoplasma cyprini n. sp. Arch. f. Potistenk.. 
52475: ; 

Woodcock, H. M. 1909. The Hemoflagellates and Allied Forms. A Treatise 
on Zoology, edited by E. Ray Lankester, Vol. 1, Fascicle I (p. 193). 


TWO NEW CASES OF POLYRADIATE CESTODES, WITH 
A SUMMARY OF THE CASES ALREADY KNOWN 


WintHrRop D. Foster 


Bureau of Animal Industry, United States Department of Agriculture 


Anomalies in adult cestodes are by no means uncommon and have 
been reported by numerous investigators in the last two centuries. 
These may be divided into those which affect only part of the worm 
and those which are characteristic of the entire strobila. Among the 
former are supernumerary proglottids, fenestrated segments, bifurca- 
tion of the strobila at the posterior end, branching at the side, forming 
a short chain of proglottids arising from a supernumerary proglottid, 
fusion of the line of separation of the proglottids through part of the 
strobila, and inversion of the usual arrangement of the sexual organs. 
Supernumerary proglottids are usually formed by the insertion of a 
smaller incompletely developed proglottid, into an otherwise normal 
proglottid (Fig. 1) or the supernumerary proglottid may be triangular 
and extend the entire length of the lateral edge of the proglottid to 
which it is attached. Fenestrated proglottids in which the central 
portion of the parenchyma is lacking are usually confined to a few 
segments, but in a few cases have involved nearly all the segments of 
the strobila. Fusion may be complete throughout a large number of 
proglottids or the line of demarcation between the proglottids may be 
obliterated only partially, giving the appearance of an excessively long 
proglottid on one side and two or more normal proglottids on the other, 
as in McCulloch’s (1913) case. 

Of more immediate interest are the cases of triradiate strobilae, in 
which the entire strobila instead of being flat or ribbon like, as in the 
normal tapeworm, has a central ridge extending uniformly through- 
out all the proglottids and giving a triradiate figure when seen in cross- 
section. This anomaly may also be combined with the anomalies affect- 
ing individual proglottids mentioned in the preceding paragraph. 
Twenty-eight such cases have been collected by Vigener (1903) and 
are summarized in the accompanying table. To this list I have added 
five more cases, one of them not hitherto reported. 


EXTERNAL ANATOMY OF POLYRADIATE CESTODES 
Analogous to the triradiate forms are those in which the tapeworm 
has more than three wings. Such forms are far less common, and 
if we except Leuckart’s case (1880), which he considered as a fusion 


8 THE JOURNAL OF PARASITOLOGY. 


of two triradiate forms, but which Barrois (1893) concludes is a 
case of simple triradiate proglottids having a simple supernumerary 
proglottid attached to one of the wings, only one case (Rosenberger, 
1903) has hitherto been reported. In Rosenberger’s case the parasite 
was pentaradiate, forming a star-shaped figure. As Rosenberger, 
however, does not specify the exact shape of the proglottids other than 
to describe them as “star shaped,” and as his observation was pub- 
lished in a journal not readily accessible to European helminthologists, 
it has been largely overlooked and the existence of strobilae having 
more than three wings is not mentioned in any of the text-books. In 
the present paper a tetraradiate proglottid of Taenia saginata is 
described. The term polyradiate is suggested as a convenient word for 
describing all cases of adult cestodes in which the strobila is formed 
of three or more wings radiating from a common axis. 

With but one exception, all triradiate cestedes in which the head 
has been recovered have been found to have six suckers instead of the 
usual four; the triradiate feature extending throughout the scolex, as 
is well seen in Vigener’s (1903) case. In Rudolphi’s (1810) case, 
however, the scolex is described as normal. Considering that the prin- 
cipal feature of this anomaly is its uniformity throughout the entire 
strobila, and that Rudolphi’s case is the only exception recorded, the 
correctness of his observation has been questioned. It is therefore 
logical to assume that a cysticercus with six suckers represents the 
larval stage of a triradiate cestode, and this view is generally accepted 
by helminthologists. Such cases are included in Vigener’s (1903) list 
of triradiate cestodes on which the present table is based. In all, 
forty-four cases of polyradiate cestodes (larvae and adults) have been 
reported. The number of individual specimens reported is, however, 
much greater, since two writers, Ziirn (1898) and Railliet (1899), 
report seeing “several” larval cestodes having six suckers, and three 
other writers each describe two or more adult specimens. 

By far the greater number of polyradiate forms are found in 
Taenia saginata, twenty-four adult cestodes of this form having been 
described. Of these twenty-four cases, two (Andry, 1741, and Brera, 
1811) are so indefinite as to be doubtful, and in four other cases the 
distinction between Taenia solium and Taenia saginata has not been 
made. These cases are assigned to T. saginata, since this parasite is 
more common than 7. solium in the regions where the cases were 
observed. Among the twenty-four cases is one pentaradiate form 
(Rosenberger, 1903) and one tetraradiate (Foster, the present paper). 


1. Vigener’s article includes a complete bibliography of all cases of triradiate 
cestodes then known. 


FOSTER—POLYRADIATE CESTODES 9 


In four species, Taenia saginata, T. solium, T. pisiformis, and T. 
coenurus, larval forms with sixsuckers have been found as well as the 
adult triradiate forms. Summarizing the cases the number of indi- 
vidual specimens reported are: 


Adult cestodes Larval cestodes 
Species Number Species Number 
Anoplocephala perfoliata ........+.0545+ 1 GUCUMYRSVCETEOIANS  o'.ccccncsciceacuseca 2 
PIENVIOLEDAGIMS TALUS). -6%5)5)<  Snie'n » olan so 1 Coenurus serialis ..................Several 
Bothriocephalus tectus ..........+25. Several USPICREGUSEOOUIS) cose vax calaeestieatves 1 
Dipylidium caninum io -Gystecercus, cellulose «02.55 03200 cass 2 
Taenia coenurus .......... 3 GYSISCETCUS PESSPOTIMS 656. oc eccnccccess 1 
Taenia echinococcus 1 Cysticercus tenuicollis ..............Several 
TSCA DUST OFMUS, vale. oes cnlen signs 08 sare ae 
SIT EaE EEE A SIREATECEGs doch a chars dave fale winris, ale wre Biel's 24 
TI OT RT CICERO RIOT IITCOEE ae RE aCe 2 
Taenia taentacformis .........ecereseces 2 


It appears from the column in the table showing the localities where 
polyradiate cestodes have been found that this anomaly is as wide- 
spread as the geographical distribution of the cestodes themselves. 
That more cases have been reported from Germany and France than 
elsewhere, is apparently due to the greater attention which has been 
given to the subject of teratological forms in these countries. 

In most cases of adult poiyradiate cestodes the scolex has not been 
recovered. Twenty-four writers have recorded cases of Taenta 
saginata, but five of them only have observed the head. This is prob- 
ably due to the fact that these specimens were recovered from the 
living hosts by vermifuges instead of at autopsy, and were therefore 
more liable to damage. The fact that cases of polyradiate cestodes 
have been found far more cemmonly in Taenia saginata than in other 
species may be due to a greater frequency of variation characteristic 
of this species, as suggested by some helminthologists, but may also 
be explained by the fact that this species, being a common parasite of 
man, is perhaps more frequently observed than any other species. 
This opinion is supported by the fact that most of the cases are 
reported by practicing physicians who have many opportunities to 
observe this species and little chance to study other species not para- 
sites of man. If this species were especially subject to this anomaly 
we should expect to find a correspondingly large number of Cysticercus 
bovis with six suckers, yet only one such case has been reported. 

A triradiate cestode usually has an unpaired wing, smaller than the 
other two wings, which are usually of nearly equal size. Sometimes 
these equal wings lie close together, as in MacCallum’s (1912) case, 
giving the worm the appearance of a normal cestode split lengthwise 
through half its width. Usually, however, the wings are thickened at 
the base so that they are separated from one another, giving a tri- 
radiate appearance. The unpaired wing may be so reduced as to form 
a mere ridge along the center of an otherwise normal parasite, as in 


10 THE JOURNAL OF PARASITOLOGY 


Jelden’s (1900) case, or it may be so well developed as to be equal in 
size and symmetry with the other wings, giving a perfectly symmetrical 
figure, as in Yoshida’s (1913) case. 

- The number and arrangement of the genital pores are subject to 
considerable variation. In most cases there is a single pore in each 
segment, located on the margin of the unpaired wing. Thus all pores 
are unilateral, an arrangement in striking contrast with the normal 
arrangement in the genus Taenia in which the pores are irregularly 
alternate. There are, however, many exceptions to this rule. In 
Bremser’s (1819) case of T. saginata, according to Rudolphi (1819) 
the genital pore was in most cases located on the unpaired wing, but 
three variations were seen: (1) genital pore not on the unpaired wing 
but on the edge of one of the paired wings; (2) genital pore on the 
unpaired wing and on the edge of one of the two paired wings; (3) 
genital pore on the unpaired wing and on each of the paired wings. 
Two proglottids each had two genital papillae on the same unpaired 
wing, one located anteriorly, the other posteriorly. In Kiichel’s (1892) 
case, according to Vigener (1903), who re-examined his material, one 
segment bore three papillae and several segments had two sexual 
openings. As a rule there was but one sexual opening to each pro- 
glottid on the edge of any one of the three wings. In Bork’s (1891) 
case although the unpaired ridge was papilliferous throughout, a super- 
numerary proglottid had a genital pore in the crevice between two of 
the wings. In Yoshida’s (1913) case of Taenia taeniaeformis (T. 
crassicollis) , “the genital pore is usually single ii each segment, situated 
on any one wing of the worm, but there are sometimes two genital 
pores lying respectively on any two wings of the segment”. In von 
Linstow’s (1892) case of Bothriocephalus tectus, the genital pores are 
all situated on the middle line of the ventral surface, the normal posi- 
tion for cestodes of this genus. 

The triradiate form in cestodes is not infrequently associated with 
the more common anomalies of supernumerary proglottids, forking, 
and fenestration. Both forking and supernumerary proglottids were 
observed by Vigener (1903) and Cattaert (1899). In the cases of 
Cattaert (1899) and Coats (1891), the worm ends in a triple fork, 
each branch forming one wing of the triradiate strobila. In McCul- 
loch’s (1913) case both fenestrated and supernumerary segments were 
frequent. The fenestration involved only mature segments, usually 
extending through two adjacent segments and following the line of the 
unpaired ridge. Asymmetrical segments were formed by the line of 
division between segments extending through only one or two of the 
three lateral wings, the opposite wing or wings being equal in length 
to the sum of these asymmetrical segments. 


FOSTER—POLYRADIATE CESTODES 11 


INTERNAL ANATOMY 


The internal anatomy of polyradiate cestodes does not as a rule, 
present any special variation from the normal except in so far as the 
arrangement of the organs is associated with the peculiar external 
form. The extra wing or wings are as fully developed internally as 
the normal part of the cestode. The uterus, having its main stem 
running through the center of the polyradiate proglottid, sends off 
lateral branches into each of the wings irrespective of their relative 
size. Thus in Jelden’s (1900) case, although the unpaired wing is here 
reduced to a mere ridge, it contains its full share of the uterine 
branches. The longitudinal excretory canals and longitudinal nerves, 
which in normal taeniae extend along each lateral margin of the 
cestode, are, in polyradiate forms, found in the same relative position 
in each of the wings. Several minor variations from the norma! have 
been recorded. Both Neveu-Lemaire (1900) and Cattaert (1899) 
observed that the transverse muscle fibers at the point where the three 
wings separate occasionally formed a partition wall separating one 
wing from the other two. In Neveu-Lemaire’s (1900) case the longi- 
tudinal canal in the unpaired ridge was larger than the other two. 
The ovaries lay in the posterior portion of the proglottid in the center 
of the “Y,” ramifying into the two equal wings but not into the 
unpaired wing. Yoshida (1913) finds that in his specimen of Taenia 
taeniaeformis, the testes are distributed throughout the three wings 
and not confined to the dorsal side as in normal specimens of this 
species. The eggs of triradiate cestodes are usually reported as normal, 
but Kiichel (1892) reports that out of ten eggs which he examined, one 
was normal, seven had eight hooklets arranged in pairs, and two had 
ten hooklets including one that was very small and incompletely 
developed. 


THE WRITER'S CASE OF A TRIRADIATE TAENIA PISIFORMIS 


Although Railliet (1892) has reported a case of Cysticercus pisi- 
formis having six suckers, no case of an adult triradiate cestode of this 
species has yet been published. The present example was found in a 
mass of tapeworms expelled by an imported collie dog held at quaran- 
tine in Athenia, N. J., and treated with a taeniafuge for tapeworm 
infestation, which had been diagnosed from « microscopic examina- 
tion. The mass of tapeworms received at this laboratory consisted of 
a great many fragments which were roughly estimated as belonging to 
from seventy-five to one hundred individuals, all of which, as far as 
examined, were of the same species, Taenia pisiformis. Although the 
entire mass was examined in a petri dish, no scolices were found. The 
identification of the species was verified by feeding experiments on a 


12 THE JOURNAL OF PARASITOLOGY 


rabbit. In this mass a number of chains of triradiate proglottids were 
found, the longest piece being 23 cm. representing the anterior half 
of the worm, except the head. In all about 52 cm. of the worm were 
recovered. 

The parasite is uniformly triradiate throughout its entire length, 
the three wings being of almost equal size and having the same angle 
between them (Fig. 1). The wings are thickened at the base, thus 
maintaining the symmetry of the figure. Owing to shrinkage from 


imm. 


Figure 1 Figure 3 


Fig. 1—A. Portion of a triradiate Taenia pisiformis, showing the twisting 
of the strobila, and consequent displacement of the papilliferous wing. B. 
Another portion of the same specimen showing a supernumary proglottid with- 
out genital pore. [Original.] 

Fig. 3.—A tetraradiate proglottid of Taenia saginata. gp, genital pore. 
[Original.] 


the formalin in which the specimen was sent, the genital pores are very 
difficult to observe in unmounted proglottids. As far as could be 
determined, however, there is but one pore to each segment, and it is 
always on the edge of the same wing. Owing to a spiral twist extending 
irregularly throughout the greater part of the strobila, the papilliferous 
wing of a given segment is seldom in line with the same wing in the 


FOSTER—POLYRADIATE CESTODES 13 


adjacent segments. Thus in Figure 1 A, while the two middle segments 
have the papilliferous wing on top, in the bottom segment it is on the 
right-hand side, while in the upper segment it is underneath. That this 
shifting of the papilliferous wing is due to the spira! twist and not to 
the fact that the pore may be on any one of the three wings, is made 
evident by finding the pores all in a straight line in those parts of the 
strobila not affected by the spiral twist. The longest proglottid was 
13 mm. long by 2.5 mm. wide. The average was 7 mm. by 3.5 mm. 
Only one supernumerary proglottid was seen (Fig. 1 B). This was 


Fig. 2.—Cross section through a gravid proglottid of a triradiate Taenia 
pisiformis, in the region of the genital pore. cc, calcareous corpuscles; clo, 
cloaca; cnf, concomitant nerve fasciculi; cp, cirrus pouch; cut, cuticle; elc, 
external longitudinal canal; ilc, internal longitudinal canal; mmnf, medullary 
nerve fasciculus; tmf, transverse muscle fibers; ut, uterus; vd, vas deferens. 
[ Original. ] 


interpolated between two normal segments and affected only one wing, 
as the line of demarcation between it and the usual triradiate proglottid 
did not extend through all three wings. The supernumerary proglottid 
had no genital pore. 

As in the other cases of triradiate cestodes, the sexual organs are 
found in all three wings. In ripe proglottids the uterus is seen to 
occupy the central portion of the body (Fig. 2), sending out branches 
into each wing. The eggs appear in all respects normal. Two longi- 
tudinal canals, the larger external and the smaller internal, appear in 


14 THE JOURNAL OF PARASITOLOGY 


each wing near the margin. ‘The relative position of these canals 
varies both in different proglottids and in the different wings of the 
same proglottid. In the drawing (Fig. 2), the canals occur laterally, 
the ventral canal being much the larger. The principal longitudinal 
nerve, the medullary fasciculus, is between these canals and the margin 
of the wing, one in each wing. In one wing two accesory fasciculi 
could be seen on either side of the medullary fasciculus. The vas 
deferens at the plane of the section drawn occupies most of the medul- 
lary layer of one of the wings (Fig. 2), and, extending into the cloaca, 
passes between the two longitudinal canals. In mature proglottids the 
testes are distributed scatteringly throughout the three wings, being 
less numerous in the region of the vas deferens. As in normal pro- 
glottids, the ovaries occupy the central portion of the posterior half, 
and send out ramifications in all directions. The arrangement of the 
transverse and longitudinal muscle fibers, calcareous corpuscles and 
all other organs are, as far as observed, no different from that seen in 
normal cestodes of this species. 


TETRARADIATE AND PENTARADIATE CESTODES 


Only one case (Rosenberger, 1903) of an adult cestode having 
more than three wings throughout its strobila, has thus far been 
reported. That such an anomaly might exist was, however, antici- 
pated by Railliet (1899), who examined a number of scolices of 
Coenurus serialis and found specimens having suckers ranging in 
number from three to ten. Railliet states in conclusion: “If the rule 
appears well established that a Taenia larva with six suckers will 
produce a worm with a triradiate chain, what malformation will arise 
from scolices having 3, 5, &, 9, and 10 suckers?” In view of the 
assumed relation between the number of suckers of the scolex and the 
wings of the strobila, it is reasonable to suppose that Rosenberger’s 
(1903) case of a pentaradiate Taenia saginata developed from a 
cysticercus with ten suckers, and that the present writer’s case of a 
tetraradiate Taenia saginata was derived from an eight-suckered 
cysticercus. 

Rosenberger (1903) received from a physician in Colorado a section 
of several proglottids of Taenia saginata having a “star-shaped” figure. 
The specimen was sent to Dr. Mohler of this bureau. Rosenberger’s © 
(1903) brief note includes a figure showing a chain of four pro- 
glottids having four equal or subequal wings radiating from a common 
center. As Rosenberger does not give the number of wings to his 
specimen and merely characterizes it as “star-shaped,” the writer asked 
Dr. Mohler for further information. Dr. Mohler stated that according 
to his recollection there were five wings radiating from a common 


FOSTER—POLYRADIATE CESTODES 15 


center, giving the star-shaped figure described, and that the fifth wing 
did not appear in the drawing since it was hidden from view by the 
other wings. No detailed study was made of the specimen and the 
number and arrangement of the genital pores was not observed. Dr. 
Mohler was under the impression that the specimen had been deposited 
in the Helminthological Collections of the United States National 
Museum, but it could not be found. In looking through the material, 
however, a tetraradiate specimen was found, described below. 

This specimen (No. 3269, Helminthological Collections, United 
States National Museum) consists of one proglottid only. The mate- 
rial was determined by Stiles in 1901 and collected the same year. 
Except for the name of the species (Taenia saginata) and the host, 


viomm. 

Fig. 4—Cross section through a tetraradiate proglottid of Taenia saginata, 
in the region of the genital pore. cl, cloaca; elc, external longitudinal canal ; 
ilc, internal longitudinal canal; mnf, medullary nerve fasciculus; wf, uterus; 
vd, vas deferens. [Original.] 


no further information is given on the label. The proglottid is 15 mm. 
long with a maximum width of 8 mm. at the posterior extremity, which 
is considerably wider than the anterior end (Fig. 3). Three of the 
wings are of fairly equal width, the fourth wing, largely concealed in 
the drawing (Fig. 3), is considerably shorter than the others. There 
is but one genital pore, placed somewhat posterior to the middle of 
the segment, on the edge of the middle one of the three equal wings. 
In cross-section (Fig. 4) the wings are seen to form an asym- 
metrical tetraradiate figure, the gravid uterus extending into each wing. 


16 THE JOURNAL OF PARASITOLOGY 


A large internal excretory canal of irregularly triangular outline is 
seen near the external edge of each wing. The smaller external canal 
appears between the internal canal and the outside edge. The uterine 
branches extending into all four wings are in most sections devoid of 
eggs and appear as large irregular lacunae in the medullary layer. A 
few eggs, however, appear in the main uterine stem (Fig. 4). The 
principal longitudinal medullary fasciculus is seen close to the wall 
of the internal canal, apparently flattened out by pressure from the 
adjacent longitudinal canal. The coiled vas deferens and the outline of 
the cloaca is seen in the middle of the three subequal! wings. 


ORIGIN OF POLYRADIATE CESTODES 


Triradiate cestodes are sometimes referred to as being a fusion of 
two normal individuals; it would seem more logical, however, to con- 
sider them as representing the fusion of one cestode with half of 
another individual, since we invariably find six suckers to these tri- 
radiate forms and not eight, which we would expect if two individuals 
were blended. If, however, there were a true fusion we should expect 
to find a line of union, which does not appear in cross-sections. More- 
over, if a cestode having irregularly alternate papillae were joined 
to another individual, we should expect to find about half of the pro- 
glottids with two genital pores and half with only one, yet it is usual 
to find only one genital pore to a segment, and that on the same wing 
throughout the strobila. From the fact that cysticerci with six suck- 
ers are occasionally found, and that oncospheres with more than six 
hooklets have been observed, it was suggested by Davaine (according 
to Railliet, 1899) that the cause of this abnormality originated in the 
egg. This view is, however, disputed by Leuckart (1880) and Railliet 
(1892), who point to the fact that in a coenurus, several of the 
scolices may have an abnormal number of suckers while the others are 
normal, yet all must have originated from the same oncosphere. 


FEEDING EXPERIMENTS WITH TRIRADIATE TAENIA PISIFORMIS 


In view of the fact that oncospheres with eight hooklets and cys- 
ticerci with six suckers have been found, it seems reasonable to expect 
that these forms would originate from a triradiate tapeworm, and 
Kiichenmeister and Ziirn (1878-81) and Railliet (1892) have sug- 
gested the advisability of feeding experiments to determine their ori- 
gin. These authors were, however, unable to carry out the suggestion 
from lack of material. 

Although the triradiate Taenia pisiformis described by the present 
writer was shipped in a solution of formalin of unknown strength, 
and kept in a 2 per cent. solution of formalin for one week after it 


. 


TABULAR LIST OF CASES OF POLYRADIATE CESTODES * 


Author 


EAMETS wale wce's 
Rudolphi .... 


Bremser ..... 
Levacher 
Siebold 

Kiichenmeister 
Kichenmeister 


Cullingworth. . 
Kiichenmeister 
4 © Zorn -..:. 
) Leukart 


Laker 
Trabut 


wees 


eee wee 
ween ee 


waeLanstow ... 


Monticelli ... 

SACEOIS® 2. sive 

Pittard (after 

Railliet) ... 

> Shennan ..... 


(eattaert ...... 
Neveu-Lemaire 


Welder o...... 
WEOHOfE «occas. 
Vigener 
Rosenberger .. 
Galli-Valerio 
MacCallum .. 
Moshida ...-; 
McCulloch 
Foster 

Foster 


ictal ie 


Date Species Locality 
1741 T. saginata? | France 
1810 Dipylidium cant- Germany? 
num 
1810 T. saginata? Switzerland? 
1819 T. saginata? Austria? 
1819 T. taentaeformis Austria? 
1841 T. saginata? France 
1853 T. echinococcus Germany? 
1855 T. coenurus Germany? 
1855 T. saginata Cape Good Hope 
1861 T. solium Germany? 
1863 C. cellulosae German 
1866 T. saginata Englan 
1870 T. saginata? France 
1873 T. saginata England 
1878- 
1881 C. cerebralis Germany 
1880 T. coenurus Germany 
1880 T. saginata Germany 
1885 T. solium Germany 
1889 T. saginata Tonkin 
1890 Anoplocephala France 
perfoliata 
1891 T. saginata Scotland 
1891 T. saginata Germany 
1892 C. pisiformis France 
1892 T. saginata East Africa? 
1892 Bothriocephalus | § S. Georgia 
tectus | Antarctic reg. 
1893 T. saginata? Italy? 
1893 T. saginata France 
1895 Bothriocephalus England? 
latus 
1898 T. saginata Scotland 
1898 C. cellulosae Germany 
1898 C. teruicollis Germany 
1899 C. serialis France 
1899 T. saginata France 
1900 T. saginata France 
1900 T. saginata Germany 
1902 C. bovis Germany 
1903 T. saginata Germany 
1903 T. saginata Colorado, U.S.A. 
1909 Coenurus serialis Switzerland? 
1912 T. saginata Canada 
1913 T. taeniaeformis Japan 
1913 T. saginata | Missouri, U.S.A. 
1915 T. saginata Un SxAr¢ 
1915 T. pisiformis Europe 


| 
' 


No. of 
Specimens 


ll aetna DOR DO eee et et et et Ot ee 


Several 
1 
1 
1 
1 
1 


Several 
Several 


Pe ph ek ek eh et et et et et om 


Appearance of 
Head 


Unknown 
Normal 


Unknown 
Missing 
6 suckers | 
Unknown | 
6 suckers 
6 suckers 
Missing 
6 suckers 
6 suckers 
Missing 
Missing 
Missing 


6 suckers 
6 suckers 
Missing 
6 suckers 
6 suckers 
6 suckers 


Missing 
Missing 
6 suckers 
6 suckers 


Missing 
Missing 
| Missing 
Unknown 


Missing 
6 suckers 
6 suckers 
7 ee PA SE ee 
suckers 
Missing 
1 case none 
| 2 cases, 6 suckers 
6 suckers 
| 6 suckers 
| 6 suckers 
Missing 
6 suckers 
Missing 
6 suckers 
Missing 


Missing 
Missing 


| 


Shape of 
Strobila 


Triradiate? 
Triradiate 


Triradiate? 
Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 


Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 


Triradiate 
Triradiate 


Triradiate 


Triradiate 
Triradiate 
Triradiate 
Triradiate 
Triradiate 


Triradiate 


Triradiate 
Triradiate 
Triradiate 
Pentaradiat 
Triradiate 
Triradiate 
Triradiate 
Tetraradiat 
Triradiate 


doubtful localities are also marked by an interrogation point. 
Strobila,” the interrogation points indicate that the specimens were so imperfectly described that it is not 
certain whether they were triradiate forms or not. 
cestodes described are larval forms and hence have no strobilae. 


*In those cases where the species has been imperfectly described so that there is some doubt whether 
the cestode seen belonged to Taenia saginata or T. solium, the cestode is assigned to the species saginata 
followed by an interrogation point, as this species is the more numerous in most countries. 
writer has failed to state the country from which the worm was collected, the locality given is that of 
the country in which he lived when the case was published (as far as could be determined). 


Where the 


These 


In the last column marked “Shape of 


The dotted lines in this column indicate that the 


18 THE JOURNAL OF PARASITOLOGY 


was received, it was determined to use some of the material for feed- 
ing experiments. The writer was encouraged in the hope that the 
vitality of the eggs would prove unaffected by the formalin, from the 
fact that on several previous occasions he had fed to rabbits, pro- 
glottids which had been shipped in formalin and had always succeeded 
in infesting the animals. In the previous cases, however, the feeding 
experiments were performed as soon as the material was received. 

A rabbit reared at the experiment station of the Bureau of Animal 
Industry was fed May, 1914, with two proglottids of the triradiate 
Taenia pisiformis already described. The rabbit died June 4, 1915. 
The postmortem revealed seven cysticerci, three of which were attached 
to the omentum, the others lying loose in the body cavity. The cys- 
ticerci were all fully grown and surrounded by a protective membrane, 
the largest cyst measuring 2 cm. long by 1 cm. in diameter. In dis- 
secting out the invaginated scolices to determine the number of suckers, 
two specimens were mutilated and the number of suckers could not be 
positively determined. There is no reason to suppose, however, that 
more than the usual number of suckers were present. The other five 
specimens were entirely normal. 

It can not be positively demonstrated that the rabbit was uninfested 
with Cysticercus pisiformis at the time it was fed. On the other hand, 
the fact that the rabbit was reared and kept in a cage until its death, 
and that as far as the writer is aware no rabbits from this source have 
been found infested with C. pisiformis unless as the result of feeding 
experiments, is very strong evidence for assuming that the cysticerci 
found resulted from the feeding experiment and not from a previous 
infestation. 

The experiment therefore failed to prove that cestode larvae with 
an excessive number of suckers are the offspring of polyradiate adults. 
On the other hand, it appears that a triradiate cestode may give rise 
to perfectly normal larvae, which presumably would develop into 
normal adults. Whether or not cestode larvae with an excessive 
number of suckers have any genetic connection with polyradiate adult 
cestodes, is a question still remaining unanswered. 


SUMMARY 


1. The term polyradiate is used to designate those cestodes whose 
strobila is uniformly divided into three or more rays or wings extend- 
ing throughout the entire strobila and radiating from a common axis, 
and whose scolices have two suckers for each of the rays present. 
Presumably the larvae of these polyradiate forms have as many suckers 
as appear in the scolices of the adults. 


ee ee 


Pe a oe ee 


FOSTER—POLYRADIATE CESTODES 19 


2. Altogether forty-four cases of polyradiate cestodes (including 
larvae) have been reported, in all but two of which the adult forms 
were triradiate. The greater number of cases are triradiate forms 
of Taenia saginata, but several species are represented and they are 
found in widely distributed localities. The greater frequency of this 
anomaly in 7. saginata is probably due to the greater chances for 
observation of this species. 

3. Of the two cases having more than three rays, one is apparently 
pentaradiate (Rosenberger’s case), and the other is tetraradiate 
(Foster’s case, the present paper); both are specimens of Taenia 
saginata. Since triradiate forms are assumed to originate from larvae 
with six suckers, it is suggested that the tetraradiate and pentaradiate 
forms originated from cysticerci having 8 and 10 suckers, respectively, 
larvae with this number of suckers having been found by Railliet 
(1899) in the case of Coenurus serialis. 


4. The view that the origin of polyradiate forms can be traced to 
the ovum, is supported by the finding of oncospheres having an exces- 
sive number of hooklets. On the other hand, this view is disputed by 
the finding of both normal and abnormal scolices in the same coenurus. 
A feeding experiment with triradiate proglottids of Taenia pisiformis 
tends to show that in this species perfectly normal cysticerci may 
result from abnormal adults. Whether or not cysticerci with an exces- 
sive number of suckers and oncospheres with an excessive number of. 
hooklets have any genetic connection with polyradiate adults, is a 
question which has not yet been solved. 


BIBLIOGRAPHY 


For complete citation of articles reported in this paper see Index Catalog 
of Medical and Veterinary Zoology, Bull. 39, Bureau Animal Industry, U. S. 
Dept. Agric. 

The following papers have appeared since the completion of that catalogue. 

MacCallum, G. A. 1912. Malformation of Taenia saginata (T. triédre}, 
Med. Rec., N. Y., 81 (12), March 23, 562-563. 

McCulloch, Hugh. 1913. Cestode Monstrosities, Am. Jour. Trop. Dis. [etc.], 
N. Orl., 1 (6), December, 453-461. 

Yoshida, S. O. 1913. Triradiate Taenia crassicollis. Parasitology, Cam- 
bridge [Eng.], 6 (3), October, 278-282. 


SOME NOTES AND EXPERIMENTS ON SARCOCYSTIS 
PEN EDA, RAILLIET + 


Joun W. Scort 


In a recent paper on Sarcosporidia encountered in the Canal Zone, 
Darling (1915) makes the interesting suggestion that the ‘Sarco- 
sporidia may be side-tracked varieties of some of the Neosporidia of 
invertebrates which have invaded the musculature of a hospitable 
though by no means definitive host and are unable to continue further 
their life cycle and escape from a compromising and aberrant position.” 
Darling thinks the high incidence of infection in cattle, sheep, swine, 
and horses favors this view, and points out the ease with which such 
animals may obtain Neosporidia in their food. In this connection the 
writer has obtained during the past year some data concerning the 
sheep sarcocyst that appear to favor a similar conclusion. Though the 
results of the experiments were negative the inferences are nevertheless 
interesting. 

A large percentage of the range sheep of Wyoming are probably 
infected with Sarcocystis tenella. Out of sixty-five sheep examined 
at different times at a local slaughter house fifty were found infected, 
or nearly 77 per cent. In some lots approximately 100 per cent. were 
infected. The parasite presented the general appearance described by 
Railliet (1895), Alexieff (1911), and others. The stages found usually 
showed an alveolar structure, a few presented the pansporoblast stage, 
but none were in the Balbiana stage. Some of the largest cysts, and 
presumably the oldest ones, had the appearance of fading out or under- 
going degeneration, and so far as one could observe from living mate- 
rial there was no indication that the parasites ever escape from the 
muscle of the host. The heart muscle and the esophagus were exam- 
ined for the cysts, but they were found much more frequently in thé 
heart. 

Since the direct observations just mentioned gave no clue to the 
life history, a preliminary series of experiments was planned to test 
various hypothetical methods of infection. Smith (1901) has suc- 
ceeded in getting a direct infection in mice by feeding muscle tissue 
from infected mice, and the first experiment was to determine if this 
method would succeed in the case of the sheep sarcocyst. While 
Smith’s experiment might account for a natural transmission of the 


* Contribution No. 2 from the Laboratory of Zoology and Parasitology, 
University of Wyoming. Experiments by aid of Adams Fund. 


SCOTT—SARCOCYSTIS TENELLA 21 


Sarcosporidia of carnivorous animals, it cannot explain transmission 
in the sheep, or in other herbivora. Dr. L. D. Swingle, formerly of the 
University of Wyoming, fed five young lambs pieces of heart muscle 
containing the sarcocysts. From the time they were born these lambs 
were fed dry feed in a dry lot and were watered from the city water 
supply which has its source in deep springs. When the writer exam- 
ined these lambs at slaughtering time not a single one was infected. 
Control jambs, kept under the same conditions, were likewise unin- 
fected. This experiment has been repeated with similar results, and it 
appears to show that the parasites in the sarcocysts are not in a trans- 
missable stage or condition, using the direct method. 

Several authors have held the view that an intermediate host is 
necessary, since the delicate structure of the spores of Sarcosporidia 
would ill adapt them to withstand the conditions outside of the verte- 
brate host. For this reason Wasielewski (1896) believes that, as in 
the case of the malarial parasite, an intermediate host is necessary in 
order to convey the parasite from one host to another. On this theory, 
Minchin (1903) suggests three possibilities in regard to infection by 
Sarcosporidia: (1) The intermediate host is a large carnivore, as the 
dog, in case of parasites like those of the pig or sheep; (2) after death 
the parasites are taken up by some carrion-feeding animal, which 
might be some vertebrate, as bird or mammal, or some invertebrate, 
as blow-fly or carrion-beetle; (3) the infection might be taken on by 
some internal parasite, for example, a flatworm or a nematode. 
Minchin regards the third explanation as unlikely, and I have fre. 
quently found lambs with no parasitic worms present, but at the same 
time infected with Sarcosporidia. He believes the second explanation 
receives some support from the extremely toxic nature of the parasites 
themselves, since this would aid in the death of the host. 

More recently Minchin and Thompson (1915) have shown that the 
rat-trypanosome passes through a cycle in the digestive tract of the rat- 
flea, and that after this period the rat becomes infected either by licking 
the flea feces or by ingesting the flea. While there is no blood-sucking 
parasite of the sheep in Wyoming that could perform a similar role 
for S. tenella and at the same time account for the prevalence of this 
parasite, the object of the following experiment was to test whether 
digestion by a carnivorous animal (coyote or dog) would render the 
Sarcosporidia contaminative. 

My second experiment, therefore, was essentially a test of Minchin’s 
first hypothesis. On August 28 a young dog was fed liberally on sheep 
hearts containing sarcocysts and placed in a wire cage where the grass 
had not been pastured. On August 31 the dog was again fed with 
infected muscle, and in the afternoon of September 2 he was taken out 


22 THE JOURNAL OF PARASITOLOGY 


of the cage. By this time feces were well scattered over the grass. 
Water was sprinkled heavily on the grass September 3 and 9 in order 
to further facilitate contamination. On September 11 two lambs, pre- 
viously kept in a dry lot, were placed in the cage and left for about 
thirty-three hours. Again October 1 two more lambs were put in the 
cage and left for twenty-four hours. A later examination showed that 
none of these lambs was infected. 

Though the experiment is not conclusive, the evidence is against 
the idea that the digestion of muscle tissue by a carnivorous animal is 
the normal method by which the Sarcosporidia of the sheep are set 
free in order that they may be accessible through its own herbivorous © 
habits. Indeed, the comparatively rare occasions en which coyotes 
and dogs obtain sheep’s flesh would hardly account for the frequency 
with which flocks are infected. 

During the summer of 1914 another experiment was tried that is 
even of more interest, and to a certain extent is a test of the second 
explanation suggested by Minchin. Two groups of lambs were used. 
Group one, consisting of twenty-one ewes and eighteen lambs, was 
allowed to graze in pasture A, where both dry and swampy condi- 
tions were present. In this pasture is located a permanent pond fed 
by seepage water, and the pond overflows except in the dry season into 
the swamp. The swampy portion of the pasture went dry about mid- 
summer. Group two, consisting of twenty-five ewes and twenty-three 
lambs, was allowed to graze in a small dry pasture B, where no water 
was present except such as fell in the frequent though scanty summer 
rains. At several times during the summer infected pieces of heart 
muscle were scattered in the pond in pasture A, and also upon the grass 
in pasture B. When the lambs were killed 55 per cent. of Group 1 
(ten out of eighteen) were found infected, and 21 per cent. (five out of 
twenty-three) of Group 2. From this result it appeared (1) that 
lambs may become infected either through water or by eating infected 
grass; or, (2) which is more probable, the infection was independent 
of the experiment. 

On the assumption that the third experiment gave positive results, 
it is hard to reconcile the infection of lambs in Group 2 with the results 
from the first two experiments. For if one does not get positive results 
by direct feeding of infected muscle, or by feeding grass contaminated 
with feces after the ingestion of infected muscle, one would not expect 
any infection where the muscle was simply scattered on the grass. 
Again, if scattering muscle containing sarcocysts on the grass or in 
water, and the consequent decay is a necessary condition for infective 
material, we are confronted with the fact that the natural death and 
decay of sheep carcasses will not account for all cases of infection. It 


SCOTT—SARCOCYSTIS TENELLA 23 


therefore appears that the infection which took place under the con- 
ditions of the third experiment was independent of the experiment. 
What other possibilities are left? 

If we assume that the sheep is the definite host of S. tenella and 
that no intermediate host is necessary, we musi conceive that the para- 
sites are set free in the blood, find their way to the exterior in the 
excretions, probably in feces or urine, and that in this way food or 
drink is contaminated. Most authors regard this as extremely improb- 
able and we have already stated that there is no evidence to show that 
the Sarcosporidia are freed in this way. Again, lambs fed along with 
ewes in a small dry lot must frequently have the hay contaminated with 
feces or urine, and yet under these conditions in the experiments no 
infection has so far occurred. On the other hand, if we consider an 
intermediate host is necessary, three possibilities arise. We may think 
of an external blood parasite as being the active agent in transferring 
the Sarcosporidia from one sheep to another; but in this case no 
external parasite appears to satisfy all the conditions, and we ought to 
get transmissions in a dry lot as well as elsewhere. Second, we may 
think of the intermediate host as feeding upon contaminated feces or 
urine and later depositing the spores on food or drink of the sheep. 
We can at once eliminate the matter of drink, for there was no water 
to be had in pasture B of my third experiment where infection 
occurred. While certain insects on this hypothesis might possibly 
account for the infection of both groups of the third experiment, it is 
probable that control lambs, confined with their ewes in an adjacent 
dry lot, would also have become infected, and we have already noted 
that there is no evidence that spores escape in the way suggested. The 
third possibility would be to assume that the intermediate host is a 
carrion-feeding animal. But no carrion-feeding vertebrate common to 
pastures A and B could be discovered. Besides, the heart muscle 
thrown into the shallow pond in pasture A sank to the bottom, where 
it was inaccessible to any sort of carrion-feeding animal, especially 
any insect, that might happen to be present. Minchin’s second sugges- 
tion, therefore, as a possible method of infection appears to be out 
of the question. 

If, however, the sheep is not the definitive host of S. tenella, but the 
presence of the parasite is more or less accidental, the results of the 
experiments given are more easily explained. First, it is evident that 
the conditions for infection are much more favorable in pasture A than 
in pasture B, though the parasites were acquired in both places. It 
is therefore well to consider further the difference between the two 
places and their similarities. In pasture B the grass was rather sparse, 
flowering plants were few in both numbers and kind, and various flies, 


24 THE JOURNAL OF PARASITOLOGY 


ants, beetles (under cattle dung), mosquitoes, bees (occasionally), 
grasshoppers, a few moths, and spiders were present. All of these 
things were present in pasture A, and, besides certain specific water 
animals, it differed from pasture B in the following particulars: There 
were more flowering plants, sedges were abundant in the swampy por- 
tions, mosquitoes and various flies were very abundant, and bees and 
moths were more frequently found. Now Erdmann (1910), Minchin 
(1912), and others look upon the Sarcosporidi2 as one group of Neo- 
sporidia, on the basis of what is already known in regard to their 
development and structure. If we accept their conclusions, and if 
according to Darling’s suggestion the Sarcosporidia are aberrant Neo- 
sporidia, and infection of herbivora is acquired by accidental ingestion 
of invertebrate hosts or by the ingestion of the droppings of such 
hests upon flowers or leaves, we can readily understand how infection 
took place in both pastures and how the percentage of infection in 
pasture A greatly exceeded that in pasture B. Further experiments 
are now in progress which will test the question of whether S. tenella 
is in reality only an aberrant form of one of the Neosporidia. 


SUMMARY 


To sum up this paper, one may say that the experiments are chiefly 
important for their negative significance. Infection with S. tenella 
failed to occur, (a) as the result of feeding infected muscle, (b) as 
the result of eating grass contaminated with feces from a carnivorous 
animal previously fed on infected muscle, and (c) by allowing infected 
muscle to decay either on dry*grass or in a pond. The apparently 
positive results of the third experiment are best explained as due to 
conditions independent of that experiment. All of the evidence favors 
the view that the sheep is not the definitive host of S. tenella, and there- 
fore is in accord with Darling’s suggestion that the muscle parasites of 
vertebrates are aberrant forms. 


REFERENCES CITED 

Alexieff, A. 1911. Sur la morphologie de la sarcosporidie de mouton 
(Sarcocystis tenella Railliet), Soc. de Biologie, 71, 397. 

Darling, S. T. 1915. Sarcosporidia Encountered in Panama, Jour. of 
Parasitol, ols: 

Erdmann, Rh. 1910. Die Entwickelung der Sarcocystis Muris in der Musku- 
latur, Sitzungsb. d. Gesellsch. Naturf. Freunde, zu Berlin. ; 


Minchin, E. A. 1903. Sporozoa: Treatise on Zoology, Ed. by E. R. 
Mankester= arp olsstaseec. 

1912. An Introduction to the Study of Protozoa, London. 

Minchin, E. A., and Thomson, J. D. 1915. The Rat-Trypanosome, Try- 
panosoma lewisi, in Its Relation to the Rat Flea, Ceratophyllus fasciatus, Ortly. 
Jour. Micr. Sci., London, 601. 

Railliet, A. 1895. Traite de zoologie médicale et agricole, Ed. 2, pp. 150-157. 

Smith, Th. 1901. The Production of Sarcosporidiosis in the Mouse by 
Feeding Infected Muscular Tissue, Jour. Exp. Med., 6. 

Wasielewski, V. 1896. Sporozoenkunde. Jena, pp. 119-127. 


7 


EGG VARIATION IN A TREMATODE SPECIES 


WILLIAM WALTER CorT 


Macalester College, Saint Paul, Minn. 


In 1903 H. B. Ward (1903a) called attention to the importance of 
the eggs in determining human entozoa. Later the same writer in a 
paper devoted entirely to the eggs of human parasites (Ward, 1908) 
emphasizes his earlier view and on page 180 comes to the following 
‘conclusion in regard to recorded observations of egg size: “The very 
existence of marked variation [in egg size] in the records of a single 
form is presumptive evidence that in the absence of errors of observa- 
tion, two or more species are confused under the single appellation.” 

In all of Looss’ extensive work on the trematodes he uses the size 
of eggs as a character of specific value, and Lithe (1909) in his sum- 
mary of the fresh-water trematodes of Germany gives the egg size in 
his account of almost every species. These and other workers on 
trematodes have generally recognized the importance of determining 
the limits of variation and the average size of ripe, normal eggs among 
trematode species. On that account the following record of a con- 
siderable variation from the species average in the size of the eggs 
from three individuals of one of the common frog lung flukes seems 
worthy of note. 

Recently while working on the anatomy of Pneumonoeces simuili- 
plexus Stafford from the lung of the leopard frog Rana pipiens more 
than two hundred eggs from ten different individuals were measured 
to determine accurately the average egg size for the species. The aver- 
age length of the eggs for this species as computed from these measure- 
ments was found to be 37.6 », and the range of variation showed a 
minimum of 34 » and a maximum of 40 ». Later, while examining 
three good-sized specimens of the same species from a single frog from 
Oshkosh, Wisconsin, I was struck by the fact that the eggs appeared 
smaller than in the forms previously examined. Measurements of two 
hundred eggs from these individuals confirmed this opinion, since the 
average egg length was found to be only 34.2 » and the limits of varia- 
tion from 30 to 37.4 ». This gives a difference between the Oshkosh 
flukes and the normal egg average length of the species Pnewmonoeces 
similiplexus of 3.4 », or a greater difference than is found between two 
distinct species of this same genus, viz., Pnewmonoeces breviplexus, 
with an average egg length of 22.5 », and Pneumonoeces longiplexus, 
with an average of 24.8 ». A study of the three individuals with the 


26 THE JOURNAL OF PARASITOLOGY 


egg variation showed that in the rest of the characters used for specific 
diagnosis they agreed with my other specimens of Pneumonoeces 
similiplexus. This observation shows that a distinct variation may 
occur within a species in a character which has proved to be generally 
constant. 

REFERENCES CITED 


Lithe, M. 1909. Parasitische Plattwiirmer. 1. Trematodes. Die Stisswas- 
serfauna Deutschlands, Heft 17. 

Ward, H. B. 1903. Data for the Determination of Human Entozoa. Trans. 
Amer. Micr. Soc., 24, 103-138. 

1908. Data for the Determination of Human Entozoa—II. Tr. Amer. 
Micr. Soc., 28, 177-202. 


SOME NEW GREGARINE PARASITES FROM 
ARTHROPODA * 


MINNIE ELIZABETH WATSON 


For several years I have been studying a number of gregarines 
parasitic in various arthropods. The literature apparently contains 
no record of most of them and consequently they are here described 
as new species. A few of the species studied were already known, 
but I am able to give new records on distribution and additional data 
on biology and life-history. Careful attention was devoted to the 
biology of the forms studied and extended experiments were con- 
ducted on life-history problems. In this paper is presented a brief 
account of these studies, which will be published in full at a later 
date. Especial attention is called here to the observations on move- 
ment in gregarines and on cyst formation. The descriptions of new 
species though concise have been worked over so carefully that it is 
thought they will be ample for accurate determinations. 


BIOLOGICAL OBSERVATIONS 


The polycystid gregarines have a septum which divides the cell 
into two or more distinct compartments, and the species described in 
this paper are all of this type. Polycystid gregarines inhabit chiefly 
the mid-intestines and intestinal diverticula of arthropods and are 
often found in large masses comprising many hundred parasites. In 
some genera the adult animals are solitary ; in others they are attached 
one behind the other. Most of the latter are biassociative, but a few 
genera occur in chains of from three to ten or twelve individuals. 

The sporonts, or adult animals, move about freely in the lumen 
of the intestines or lie inert between the lobes. The trophozoites, or 
young individuals, live either entirely within the epithelial cells, as 
in the family Stenophoridae, or attached to the free ends of the cells 
by means of variously shaped epimerites, globular in the genus Gre- 
garina. When a trophozoite has absorbed sufficient nourishment from 
the host cell, either directly through its walls or by means of the 
epimerite, it breaks forth from the shriveled cell and becomes a spo- 
ront, living free in the intestine, and the useless epimerite, if present, 
is gradually lost. The animal now receives its food entirely by absorp- 
tion of the digestive juices direct from the host intestine. 


* Contributions from the Zoological Laboratory of the University of Illi- 
nois, under the direction of Henry B. Ward, No. 48. 


28 THE JOURNAL OF PARASITOLOGY 


Movement of the free individuals takes place by a gradual progres- 
sion and by bending of any part of the deutomerite. There are two 
structures necessary for motion. Running crosswise in the outer 
part of the endocyte is a delicate network of fibrillae, the myonemes, 
sometimes seen with a rather low power in individuals nearly devoid 
of protoplasm. In the outer portion of the epicyte, the outside layer 
of the body, there are found very fine longitudinal striations visible 
only with an oil-immersion lens. In the furrows between these stria- 
tions there are minute pores (Schewiakoff, 1894) through which a 
gelatinous material is exuded. The animal progresses by contracting 
a few myonemes on that side of the body which happens to be ven- 
tral and this causes a minute undulatory motion against the slide. At 
the same time the animal secretes mucus, which enables it to move 
forward against friction, much as a slug moves forward by a wave- 
like motion on its ventral side. The trail of mucus left on the slide 
is the now useless material which has gradually been pushed back- 
ward through the longitudinal furrows by the progressing animal. 
Bending movement is effected by a contraction and expansion of the 
myonemes in any part of the deutomerite. 

As the beginning of cyst-formation, two individuals, either asso- 
ciative or solitary, commence to revolve in a large circle. If the 
animals are solitary, an individual is drawn into the vortex of one 
which has started to revolve alone. If of a biassociative type, the 
two commence to revolve together. The spiral gradually becomes 
smaller as they continue in motion and the animals come to lie in 
contact laterally. Motion still continues and becomes rotary. As the 
two sporonts are forming a compact sphere, a thick, transparent cov- 
ering is being laid down on the outside of the cyst which consists of 
as many thin layers of gelatinous material exuded from the posterior 
end of the moving animals as there are rotations before the ani- 
mals come to rest. The fully formed cyst, sometimes still rotating, 
now passes from the mid-intestine of the host to the rectum and is 
given out with the feces. 

Finding suitable moisture, the cyst develops within from 24 to 
about 48 hours with the formation and growth in many genera of as 
many as fourteen enormously long spore ducts. Each sporont breaks 
up into gametes, the gametes from one sporont uniting when fully 
formed with those from the other to form zygotes; and when the 
resultant spores have become mature they are forced out violently 
through the spore ducts into the surrounding medium. They become 
scattered and if accidentally eaten by an insect of the same species 
as the host, the outer spore wall is dissolved in the intestine, releas- 
ing eight active sporozoites. The latter pass to the epithelial cells 


WATSON—NEW GREGARINE PARASITES 29 


and either become attached or completely embedded, when the life- 
cycle begins anew. 

There is evidence to indicate that auto-infection may occur by 
the cysts ripening in the intestine, and this would account for the 
enormous number of parasites often found within a single host. 

The arthropods from which the parasites were taken include the 
Diplopoda, Orthoptera and Coleoptera, and the species are grouped 
in this order in the text. The gregarines described are members of 
the following genera: Amphoroides, Steinina, Stenophora, Gregarina, 
and a new genus which is here designated Leidyana. 


GREGARINES IN THE DIPLOPODA 


Infection in the diplopods is fairly heavy and about three fourths 
of the individuals examined at Urbana were parasitized. Parasites 
were abundant in the early spring as well as in the fall. Some species 
were nearly always found to be infected, others never. 


Stenophora diplocorpa n. sp. (Fig. 1): Sporonts solitary, elon- 
gate. Maximum length 360u, width 15. Ratio, length protomerite: 
total length :: 1:16 to 1:25. Ratio, width protomerite: width deu- 
tomite :: 1:2 to 1:3. Protomerite dome shaped, widest at posterior 
margin and as wide as long. Slight constriction at septum. Deuto- 
merite slender, elongate, incompletely divided into two nearly equal 
parts by a crosswise constriction, widest just anterior to this constric- 
tion. Cylindrical behind the constriction and broadly rounded at 
posterior end. Protomerite nearly transparent, deutomerite pale tan, 
not opaque. Nucleus visible in vivo, situated just behind constriction 
in the deutomerite, spherical, and containing one karyosome. Cyst 
and spores unknown. 

Taken at Urbana, Illinois. Host: Euryurus  erythropygus 
(Brandt). Habitat: intestine. 


Stenophora impressa n. sp. (Fig. 2): Sporonts solitary, ellipsoidal. 
Maximum length 375y, width 48. Ratio, length protomerite: total 
length ::1:12. Ratio, width protomerite: width deuteromite : : 1: 2.3. 
Protomerite conical, dilated in posterior half, as wide as high. An 
apparent pore at anterior end. Constriction at septum not deep. Deu- 
tomerite ellipsoidal, widest through central part, posterior extremity 
blunt or rounded. Endocyte of protomerite nearly transparent, of 
deutomerite opaque. Nucleus spherical with one large karyosome. 
Cysts spherical, 160% in average diameter. Spores not known. 

Taken at Urbana, Illinois. Host Parajulus impressus~ (Say). 
Habitat: intestine. 

Stenophora lactaria n. sp. (Fig. 3): Sporonts solitary, elongate, 
ellipsoidal. Maximum length 480%, maximum width 39. Ratio, 


30 THE JOURNAL OF PARASITOLOGY 


length protomerite: total length :: 1:10to 1:16. Ratio, width proto- 
merite: width deutomerite : : 1: 1.2. Protomerite conical, dilated above 
base and tapering to a point. An apparent pore at apex. As broad 
as high. Constriction at septum. Deutomerite ellipsoidal, widest in 
anterior third, tapering to an acute, rounded extremity. Endccyte of 
protomerite nearly transparent, of deutomerite opaque. Nucleus ellip- 
soidal, twice as long as wide. Cysts spherical, 150 to 170y in diameter. 
Spores not known. 

Taken at Urbana, Illinois. Host: Callipus lactarius (Say). Habi- 
tat: intestine. 

Amphoroides calverti (Crawley) (Fig. 4): Sporonts solitary, 
elongate. Maximum length 1670y, average length 1400y, average 
width 120. Ratio, length protomerite: total length :: 1:47. Ratio, 
width protomerite: width deutomerite : : 1:2.5 to 1:3. Protomerite 
greatly compressed in sporonts, shallow, five times as wide as high. 
Deep crater within top. Constriction at septum sharp and deep. Deu- 
tomerite elongate, widest in anterior third, tapering to a sharp point. 
Endocyte of protomerite tan in color, not dense; of deutomerite 
opaque, white. Nucleus small, spherical, not visible in vivo. Myocyte 
well developed. Cysts spherical, averaging 380 in diameter. Dehis- 
cence by simple rupture. Spores not known. 

‘Laken at Urbana, Illinois. Host: Callipus lactarius (Say). Habi- 
tat: intestine. 

This species was described by Crawley (1903a) as Gregarina cal- 
verti, but the elongate shape of the sporonts, great size, dehiscence of 
the cysts by simple rupture, and the fact that all the animals are soli- 
tary prove that the species is not a member of the genus Gregarina. 
I place it in the genus Amphoroides because of the crateriform 
protomerite. 


GREGARINES IN THE COLEOPTERA 


The following nine species have been found in beetles and beetle 
larvae in the two general localities mentioned. In no instance has a 
complete life-history been established, but the generic position is deter- 
mined beyond doubt by known characters. The members of the genus 
Gregarina are superficially very similar, but a close inspection yields 
points of difference sufficient to indicate the individuality of each 
species. While the primites of several species are similar, the satel- 
lites are dissimilar and afford one means of differentiation, The rela- 
tive sizes of the species and the color and density of the protoplasm 
afford other means of identification. The visibility of the nucleus is 
important in identification. The literature has been carefully investi- 
gated in the anticipation that some of the species, especially thase in 


WATSON—NEW GREGARINE PARASITES 31 


the Elateridae and the Tenebrionidae had been previously described. 
All are, however, new. 

Gregarina katherina n. sp. (Fig. 5): Sporonts biassociative, ellip- 
soidal. Length of associations 96 to 150u. Sporonts 45 to 70 long, 
20 to 34 wide. Ratio, length protomerite: total length primite 
1:6. Ratio, width protomerite: width deutomerite :: 1:7. Proto- 
imerite of primite dome shaped, of satellite flattened. Deutomerite 
ellipsoidal. Nucleus spherical, one large karyosome. Epimerite large, 
sessile, a hyaline knob. Cyst and spores not known. 

Taken at Oyster Bay, Long Island, N. Y. Host: Coccinella novum- 
notata Herbst. Habitat: intestine. 


Gregarina barbarara n. sp. (Fig. 6): Sporonts biassociative, ovoi- 
dal to subspherical. Length of association (average) 250. Sporonts 
(primites) average 145y long, 90u wide. Ratio, length protomerite: 
total length primite :: 1:6. Ratio, width protomerite: width deuto- 
merite :: 1:2.2. Protomerite hemispherical in primite, flattened in 
satellite, six times as wide as high, deutomerite ovoidal in primite, 
widest part in central region. Deutomerite of satellite widest in ante- 
rior third, no constriction at septum, contour here perfectly smooth. 
Nucleus small, spherical, with one karyosome. Body practically trans- 
parent. Cyst and spores not known. 

Taken at Oyster Bay, Long Island, N. Y. Host: Coccinella sp. 
Habitat: intestine. 

Gregarina globosa n. sp. (Fig. 7): Sporonts biassociative, sub- 
globose. Length of associations 4354. Length of sporonts 260p, 
width 180u. Ratio, length protomerite: total length ::1:8.6. Ratio, 
width protomerite: width deutomerite :: 1:2.4. Protomerite hemi- 
spherical, broadest at base, no constriction at septum. Deutomerite 
nearly spherical. Protoplasm dense, dark gray to black in primite, 
lighter in satellite. Nucleus spherical. Cyst and spores not known. 

Taken at Urbana, Illinois. Host: Coptotomus interrogatus Fab. 
Habitat : intestine. 

Gregarina monarchia n. sp. (Fig. 8): Sporonts biassociative, elon- 
gate cylindrical. Length of associations 570, width 130y. Ratio, 
length protomerite: total length :: 1:7. Ratio, width protomerite: 
width deutomerite :: 1:1.2. Protomerite subspherical, widest 
through middle portion, constriction at septum. Deutomerite clongate 
cylindrical, equal in width throughout, broadly rounded posteriorly. 
Deutomerite dense, black in transmitted light. Protomerite -nearly 
transparent. Nucleus not visible in vivo. Cyst and spores not known. 

Taken at Urbana, Illinois. Host: Pterostichus stygicus Say. Hab- 
itat: intestine. 


32 THE JOURNAL OF PARASITOLOGY 


Gregarina intestinalis n. sp. (Fig. 9): Sporonts biassociative, 
broadly ellipsoidal. Length of associations 3204. Maximum length 
of sporonts 160”, maximum width 80p. Ratio, length protomerite: 
total length :: 1:5. Ratio, width protomerite: width deutomerite : : 
1:2. Protomerite subspherical, widest through middle portion, deep 
constriction at septum. Deutomerite broadly ellipsoidal, protoplasm 
dense, dark gray. Nucleus not visible in vivo. Cyst and spores not 
known. 

Taken at Urbana, Illinois. Host: Pterostichus stygicus Say. Habi- 
tat: intestine. 

Gregarina gracilis n. sp. (Fig. 10): Sporonts biassociative, elon- 
gate ellipsoidal. Maximum length of associations 3/04; maximum 
length of sporonts 190%, maximum width 75y. Ratio, length proto- 
merite: total length :: 1:8. Ratio, width protomerite: width deuto- 
merite :: 1:2. Protomerite hemispherical. Deutomerite ciongate 
ellipsoidal. Color gray. Nucleus not visible in vivo, spherica!, small, 
with one karyosome. Cysts average 90u in diameter. Spores not 
known. 

Taken at Urbana, Illinois. Host: larvae of Elateridae. Habitat; 
intestine. 

Gregarina tenebrionella n. sp. (Fig. 11): Sporonts biasseciative, 
subglobose, very small. Maximum length of association, 140, aver- 
age length 125». Ratio, length protomerite: total length :: 1:4. 
Ratio, width protomerite: width deutomerite :: 1:1.7. Protomerite 
dome shaped, deutomerite nearly spherical in primite, ellipsoidal in 
satellite. Nucleus small, spherical. Protoplasm gray. Cyst and 
spores not known. 

Taken at Urbana, Illinois. Host: larvae of Tenebrionidae. Habi- 
tat: intestine. 

Gregarina fragilis n. sp. (Fig. 12): Sporonts biassociative, ellip- 
soidal. Length of associations 200u. Maximum length of sporonts 
110u, maximum width 60u. Ratio, length protomerite: total length 
primite :: 1:5. Ratio, width protomerite: width deutomerite : : 1:2. 
Protomerite dome shaped, cylindrical in posterior third. Protomerite 
of satellite same shape but slightly flattened anteriorly. Deutomerite 
ellipsoidal. Nucleus small, spherical, with one karyosome. Body prac- 
tically transparent. Cyst and spores not known. 

Taken at Urbana, Illinois. Host: Coccinella sp. Habitat: 
intestine. 


Stemina rotunda n. sp. (Fig. 13): Sporonts solitary, globose. 
Maximum length 250u, maximum width 130u. Ratio, length proto- 
merite without epimerite: total length : : 1:2.3. Ratio, width proto- 
merite: width deutomerite :: 1:1.1. Protomerite conoidal, dilated 


WATSON—NEW GREGARINE PARASITES 33 


at beginning of posterior two thirds, constricted at septum. Proto- 
merite densest in posteri®r half. Deutomerite spherical to obovate, 
posterior end either rounded or slightly pointed. Nucleus large with 
one large karyosome in young, with many chromatic bodies in adult. 
Endocyte light brown. Epimerite spherical, hyaline, persistent on 
large animals free in lumen of intestine. Cyst and spores not known. 

Taken at St. Joseph, Illinois. Host: Amara angustata Say. Habi- 
tat: intestine. 


GREGARINES IN THE ORTHOPTERA 


Five of the following species are new, one representing a newly 
created genus. New distribution records and new measurements are 
given for three species which are already known in the literature. 

Gregarina nigra n. sp. (Fig. 14) —Sporonts biassociative, cylindri- 
cal. Maximum length of associations, 1000p. Maximum length of 
sporonts 530u, maximum width 180. Ratio, length protomerite : total 
length primite :: 1:4. Ratio, width protomerite: width deutomerite 
:: 1:1.4. Protomerite a truncate cone angular at the free corners. 
Width equal to height. Widest at base, no constriction or a very 
slight constriction at septum. Protomerite of satellite scarcely flat- 
tened. Deutomerite cylindrical, broadly rounded posteriorly. Endo- 
cyte black. Nucleus not visible in vivo, spherical, containing many 
small karyosomes. Cysts and spores not known. 

Taken at Urbana, Illinois. Hosts: Melanoplus femur-rubrum 
(deGeer) ; M. differentialis (Uhler) ; Encoptolophus sordidis (Bur- 
meister). Habitat: intestine. 

Gregarina stygia n. sp. (Fig. 15): Sporonts biassociative, obese. 
Maximum length of associations 360y, length sporonts 180. Primite 
and satellite of approximately the same length. Maximum width of 
primite 100u. Ratio, length protomerite: total length primite :: 1:6. 
Ratio, width protomerite: width deutomerite :: 1:1.6 to 1:2. Pro- 
tomerite hemispherical in primite, flattened in satellite. Deutomerite 
of primite broadly ellipsoidal, nearly as wide as long ; of satellite wid- 
est in anterior half, tapering slightly. Nucleus small, spherical. Endo- 
cyte dark tan but not dense, nucleus visible in vivo in both primite 
and satellite. Sarcocyte thicker in both protomerites than in the deu- 
tomerites. Trophozoite with a simple, small, knobbed epimerite. 
Cysts 150 in diameter. Spores not seen. 

Taken at Cold Spring Harbor, Long Island, N. Y. Host: Ceu- 
thophilus stygicus (Scudder). Habitat: intestine. j 

Gregarina galliveri n. sp. (Fig. 16): Sporonts biassociative, maxi- 
mum length of associations 5901; maximum length of sporonts 300u, 
width 1304. Ratio, length protomerite: total length primiten 27) Be5 


34 THE JOURNAL OF PARASITOLOGY 


Ratio, width protomerite: width deutomerite :: 1.1:1. Protomerite 
flattened, broad and low. Three times as wide as high. Deutomerite 
of primite vase shaped, constricted at top, widening in posterior half. 
Deutomerite of satellite subspherical to ovoidal. Endocyte very dense 
in both protomerite and deutomerite, dark brown in color. Nucleus 
small, spherical, not visible in vivo. Cysts spherical, 350 in average 
diameter. Spore ducts numerous. Spores not seen. 

Taken at Oyster Bay, Long Island, N. Y. Host: Gryllus abbrevi- 
atus Serv. Habitat: intestine. 

Gregarina illinensis n. sp. (Fig. 17): Sporonts biassociative, 
elongate cylindrical. Maximum length of associations 1100u; length 
sporonts 550, width 180. Ratio, length protomerite: total length 
primite :: 1:5. Ratio, width protomerite: width deutomerite 
1:1.1 to 1.5. Protomerite dome shaped, slightly constricted at sep- 
tum. Deutomerite elongate cylindrical, broadly rounded behind. Pro- 
tomerite of satellite cupped at top for insertion of posterior end of 
primite. Nucleus large, spherical, with many small chromidial bodies. 
Endocyte dense, black in both protomerite and deutomerite. Cysts 
and spores not recovered from the host. 

Taken at Urbana, Illinois. Host: Ischnoptera pennsylvanica 
(deGeer). Habitat: intestine. 

Gregarina achetae-abbreviatae Leidy (Fig. 18): Sporonts biasso- 
ciative, obese. Maximum observed length 500”; average sporonts 
450u long, 2254 wide. Ratio, length protomerite : total length primite 
: : 1:3. Ratio, width protomerite : width deutomerite : :1:1.1. Pro- 
tomerite hemispherical to subglobose, width twice the height. Slight 
constriction at septum. Deutomerite stout bodied, nearly as wide as 
long. Widest at shoulder where it is very little wider than protomerite. 
Posterior end truncate. Epimerite undescribed. Endocyte dense in 
deutomerite, less so in protomerite. Nucleus not visible in vivo and 
not seen. Cysts spherical, 250u in average diameter. Spore ducts 
two to five in number, of maximum length 10004. Spores barrel 
shaped, 4.5 2.25p. 

Taken at Haverford, Pa., and Urbana, Illinois. Host: Gryllus 
abbreviatus Serv. Habitat: intestine. 

Gregarina rigida (Hali) Ellis (Fig. 19): Sporonts biassociative, 
stout bodied. Maximum length of associations 1425y, average length 
550u. Sporonts 250 to 750y long, 130 to 210u wide. Ratio, length 
protomerite : total length of primite :: 1:3 to 1:6. Ratio, length 
protomerite : total length satellite :: 1:5 to 1:16. Ratio, width pro- 
tomerite : with deutomerite : : 1: : 1.4. Protomerite somewhat flattened, 
width sometimes three times the height, generally less. Constriction 
at septum more or less indistinct. Deutomerite cylindrical or barrel 


WATSON—NEW GREGARINE PARASITES 35 


shaped, little wider than protomerite, ending in a broadly rounded or 
flattened square- corneredvextremity. Endocyte very dense and brown- 
ish yellow in deutomerite, tan in protomerite. Epimerite a small, 
spherical, hyaline knob. Cysts yellow-orange, 300 in av erage diame- 
ter, spore ducts short, ten or more in number. Spores extruded in 
chains, barrel shaped, 5X8n. 

Taken at Lincoln, Neb., Colorado Springs, Colo., and Urbana, Ill. 
Hosts: Melanoplus femur-rubrum (deGeer); M. differentialis 
(Uhler); M. coloradensis (?); Encoptolophus sordidis (Burm.) ; 
Schistocerca americana Burm.; Melanoplus bivitattus (Say); and 
Hesperotettix pratensis Scudder Habitat: intestine and pyloric caeca. 

This species was first described by Hall (1907) as Hirmocystis 
rigida. Crawley (1907) found it shortly after and named the species 
Gregarina melanopli. Ellis (1913) changed the name to Gregarina 
rigida. 

Leidyana solitaria n. gen., n. sp. (Fig. 20): Sporonts solitary, 
cylindrical. Maximum length 500n, maximum width 160u. Ratio, 
length protomerite: total length :: 1:5 to 1:7. Ratio, width proto- 
merite: width deutomerite :: 1:1.3 to 1:1.7.  Protomerite conical, 
dilated in middle portion, constricted deeply at septum. Protomerite 
slightly wider than high in adults. Deutomerite cylindrical to elongate 
ellipsoidal, sometimes tapering, rounded posteriorly. Endocyte of pro- 
tomerite pale tan, translucent, of deutomerite very dense, black in 
transmitted light, the two parts very plainly demarked, nucleus not visi- 
ble in vivo, spherical, with one or two small karyosomes. Epimerite a 
large, globular, hyaline knob on a short, slender stalk. Cysts spherical, 
350 in diameter (including the transparent covering). Dehiscence by 
spore ducts one to twelve in number. Spores given out in chains, 
barrel shaped, 3 by 6x. 

Taken at Cold Spring Harbor and Oyster Bay, L. I., N. Y., Haver- 
ford, Pa., and Urbana, Ill. Host: Gryllus pennsylvanicus Burm. 
Habitat: intestine. 

This species was described by Crawley (1907) under the name 
Stenophora erratica. The mode of cyst dehiscence, however, precludes 
the possibility of its belonging to the family Stenophoridae. I have 
placed it in a new genus under the family Gregarinidae, characterized 
as follows: Leidyana n. gen. Sporonts solitary, epimerite a simple 
globular knob, dehiscence by spore ducts, spores doliform. 

I should restrict the genus Gregarina to biassociative sporonts only, 
the other characters being identical with those of the new genus. 


LITERATURE CITED 


Crawley, Howard. 1903. List of Polycystid Gregarines of the United 
States. Proc. Acad. Nat. Sc., Philadelphia, 55: 41-58; 3 pl. 


36 THE JOURNAL OF PARASITOLOGY 


1907. The Polycystid Gregarines of the United States (Third Contribu- 
tion). Proc. Acad. Nat. Sc., Philadelphia, 59: 220-8; 1 pl. 

Ellis, M. M. 1913. A Descriptive List of the Cephaline Gregarines of the 
New World. Tr. Am. Micr. Soc., 32: 259-96; 4 pl. 

Hall, M. C. 1907. A Study of Some Gregarines with Especial Reference 
to Hirmocystis rigida, n. sp. Univ. Studies (Lincoln, Neb.), 7: 149-74; 1 pl. 

Schewiakoff, B. 1894. Ueber die Ursache der fortschreitenden Bewegung 
der Gregarinen. Zeit. wiss. Zool., 58: 340-54; 2 pl. 


EXPLANATION OF: PLATES t AND: 2 


Fig. 1—Sporont of Stenophora diplocorpa n. sp. from camera lucida draw- 
ing of the original. 
Fig. 2—Sporont of Stenophora impressa n. sp. 
Fig. 3.—Stenophora lactaria n. sp. 
Fig. 4—Amphoroides calverti (Crawley). 
Fig. 5—Gregarina katherina n. sp. 
Fig. 6—Gregarina barbarara n. sp. 
Fig. 7.—Gregarina globosa n. sp. 
Fig. 8.—Gregarina monarchia n. sp. 
Fig. 9—Gregarina intestinalis n. sp. 
Fig. 10.—Gregarina gracilis n. sp. 
Fig. 11.—Gregarina tenebrionella n. sp. 
Fig. 12.—Gregarina fragilis n. sp. 
Fig. 13.—Steinina rotunda n. sp., a trophozoite with epimerite, 
Fig. 14.—Gregarina nigra n. sp. 
Fig. 15.—Gregarina stygia n. sp. 
Fig. 16—Gregarina galliveri n. sp. 
Fig. 17.—Gregarina illinensis n. sp. 
Fig. 18.—Gregarina achetae-abbreviatae Leidy. 
Fig. 19—Gregarina rigida (Hall) Ellis. 
Fig. 20.—Leidyana solitaria n. gen., n. sp. 


1 


PLATE 


Seeeeee 


SAHOQUOOUCC Es 


PNEUMONYSSUS FOXI, NOV. SP. 


AN ARACHNOID PARASITIC IN THE LUNG OF 
A MONKEY (MACACUS RHESUS) 


Frep D. WEIDMAN 


On March 7, 1914, an adult male Macacus rhesus died in the Phila- 
delphia Zoological Gardens with a subacute catarrhal colitis. Its 
lungs contained, in addition, sixteen to twenty small lesions about 
equally divided between the two organs. They were nodular, 2 to 5 
mm. in diameter, were situated immediately under the pleura and 
slightly elevated above the same. The smaller ones were firm 
throughout, the larger ones with softer umbilicated centers and indu- 
rated edges. In some of the lesions smaller hard points were to be 
made out, suggesting a conglomerate lesion. When fresh they were 
pink or gray and upon incision found to contain granular gray mate- 
rial with granular gray walls. Upon scraping out the centers of the 
lesions and examining the contents, the parasitic character of the 
lesions was at once determined. 

Microscopically, the sections of the lesions exhibit sections of an 
arthropod, lying in granular necrotic material, together with brown- 
ish black, extremely finely granular detritus (excrement). Around 
this there is a slight round-cell infiltrate, very poor in leukocytes and 
in close relation to a bronchus. A thick fibrous wall of very young 
type surrounds the whole. 

The material used in making the following description was 
obtained by gently scraping out the interiors of two of the pulmon- 
ary lesions. This yielded, from one sac, eleven females and one male; 
from the other, five females. Free larvae or ova were not found. 
The parasites had been fixed in situ by formaldehyd 4 per cent., 
followed by washing in water and hardening in alcohol. Some speci- 
inens were teased, the remainder preserved entire and examined first 
in a watery medium, followed by clearing in glycerin-alcoho! mix- 
ture, or Farrant’s medium. During these studies it was found that 
the delicate membranes of the caroncle could only be satisfactorily 
examined prior to prolonged immersion in glycerin or Farrant’s 
medium, both of which had a marked tendency to produce shrinkage 
of the same. Finer structures, such as hairs and plates, were only 
successfully determined after clearing for several days and examina- 
tion with the oil immersion lens. All of the specimens proved to be 


38 THE JOURNAL OF PARASITOLOGY 


clearly of the same species. None were observed living, the material 
having been submitted subsequent to fixation. 

Grossly, they were barely visible as minute, ovoid, glistening, 
opaque, white or faintly yellowish bodies. 


THE FEMALE 


Average females, unruptured and not much distorted or flattened 
by pressure measure as follows :— 


Pubescent female 0.850 & 0.450 mm 
Ovigerous female 0.960 « 0.560 mm 
0.940 & 0.500 mm 
0.750 * 0.400 mm 


This gives an average of 0.875 * 0.478 mm. They are ovoid, the 
body not divided or constricted, the greatest width lying immediately 
behind the last pair of legs. In none of the specimens, and this also 
applies to the male, can the outline or even site of internal organs be 
made out. At most the ovum is discoverable, with occasionally the 
outlines of a folded embryo, in a ruptured specimen. 

In one of the teased specimens dorsal and ventral plates of muscle 
can be made out. Each lies median, between the two middle pairs of 
legs. The ventral is much the heavier. It sends many fasciculi later- 
ally to the coxae, a few radiate anteriorly to the capitulum and several 
posteriorly to the ends of a long, special muscle band extending trans- 
versely between the last pair of coxae. Fasciculi also radiate from the 
dorsal plate, a few anteriorly and many laterally to the coxae. There 
are other bands extending circumferentially around the posterior body 
half, but the condition of the specimen illustrating the muscular 
arrangement does not permit exact description here. 

The head is continuous with the body both dorsally and ventrally. 
There are no eyes. The rostrum projects slightly beyond the general 
body contour, is triangular, the apex rounded, the lateral edges curled 
dorsally, but curving quickly ventrally again at the apex. Jn this 
way a short, broad longitudinal groove is produced over the dorsum 
of the rostrum. The hypostome is quadrilateral, save for a slight 
median anterior marginal peak, a little longer than broad and does 
not project beyond the rostrum. Its surface is finely pebbled like 
morocco leather. A shallow longitudinal median furrow extends 
through almost its whole length, stopping just short of the anterior 
margin. In the depths of this groove 8 to 13 short blunt teeth are 
noted, set in oblique manner with their points directed anteriorly 
and ventrally. On either side of the furrow transverse or slightly 
oblique muscle bundles are seen under the cuticle, which appear to 
pass anteriorly and median to the bases of the teeth. Midway between 
the furrow and the lateral hypostomal border, and well short of the 


WEIDMAN—PNEUMONYSSUS FOXI, NOV. SP. 39 


anterior margin of the hypostome, a hair is seen on each side. In 
addition two small papillae are present on each side of the furrow 
on the anterior margin of the hypostome. 

The mouth parts are markedly retracted, barely projecting beyond 
the hypostome and not at all beyond the rostrum. The palpi lie 
dorsal and lateral to the mandibles. They consist apparently of 
three segments, of which the terminal is best seen, subspherical and 
capped by a stout, moderately long hair. The mandibles are che- 
late, lie within a sheath in which they are so far retracted as to be 
generally invisible. At times the two pointed, untoothed fingers of 
each, one shorter than the other, may barely extend beyond the mouth 
parts. 

There are four pairs of legs, the first two geniculate. The first 
pair is close to, and its proximal segment (coxa) fused with, the 
capitulum. The second pair is close to the first pair, with no inter- 
coxal space. The third pair is but a short distance, perhaps the width 
cf a coxa, behind the second, and the fourth pair is at a similar 
distance from the third. There is no special interval separating the 
first and last two pairs of legs. The first three legs are subequal 
in length (0.22 mm.), the last one a little longer (0.25 mm.).* No 
legs are as long as the body width. Each leg has six articles, although 
at first glance they may appear to have seven or eight. This is due 
to the presence of the “fehlenden” muscular insertion of Winkler, 
which produces a ring simulating the division line between two seg- 
ments. This ring appears constantly in the tarsus, is well marked 
constantly in the femora of the two middle pairs of legs and more 
faintly and only fairly constantly in the femora of the first and 
tourth pairs of legs. The tibia and patella are of about the same 
size, much shorter and heavier than the tarsus. The femur is at 
least twice as long as broad. The trochanter and coxa are much 
heavier than any of the other segments and irregularly pyramidal 
in form. The coxa is fully twice as broad as long, its posterior 
wall long, its anterior shortened by half in the last pair of Jegs and 
almost to nil in the three anterior pairs. Its ventral surface bears, 
distally, a cuneiform process which bears against a special chitinous 
plate of the trochanter. The cuticle over all segments bears long, 
stiff, straight or slightly curved hairs, their insertions surrounded 
by a low rounded ridge. They are most numerous on the distal 
segments and scarce but constant on the proximal ones. Spurs are 
constant on the three distal segments, on the other three variable. 


* An ovigerous female 0.75 & 0.40 mm. is used for all measurements unless 
specially noted. 


40 THE JOURNAL OF PARASITOLOGY 


As a rule there are none on the coxae or trochanter; if present 
at all it is generally the first two pairs of legs which bear 
them. 


Each leg has a special terminus. The first pair has two arched, 
subparallel, fairly heavy dorsal claws. Their extremities overhang 
or touch a single, median, compound-curved, ventral projection with 
a sharp point, which has the character of neither claw nor spine, 
appearing more like a chitinous, elongated, pointed tongue. This leg 
has no caroncle, the base of the claws being set directly into the sub- 
stance of the tarsus. 


The second pair is terminated by a short, broad caroncle measur- 
ing about twice as long as broad. It is pyriform, the handle inserted 
into the tarsus, the ventral and lateral distal parts open to permit 
protrusion of the claws, the whole now coming to resemble a hood 
with the opening directed ventrally. It is delicately membranous 
distally, and heavier proximally. Two strong claws are attached to 
its dorsum internally. These are parallel at their origins deep in 
the caroncle, but at their middles become strongly bent laterally, 
their tips thus coming to diverge and often to project laterally beyond 
the margin of the caroncle. The details in the depths of the caron- 
cle (the handle) are uncertain. It appears that a median ventral, 
blunt, chitinous tooth extends from it parallel to the ventral car- 
oncular wall and perhaps continuous with it. It may be straight or 
lightly curved. The margin of the proximal border of the opening 
shows a short, pointed, median projection. 


The third pair of legs is terminated by a double hooked caroncle 
precisely like that of the second pair. 


The fourth pair also has hooked caroncles built on the same gen- 
eral plan as the preceding, but they are much more slender, meas- 
uring about three times as long as broad. Its hooks, too, are much 
more delicate, more gracefully curved and do not, in the specimens 
studied, extend beyond the cavity. They are supplemented by a 
smaller, straighter pair deep in the caroncle, which is not always vis- 
ible, probably from retraction. The median tongue or tooth is again 
seen here, but is much smaller, and, too, the median, marginal peak 
is seen at the proximal border of the caroncular opening. 


The cuticle appears for the most part to be soft. It has a peb- 
bled appearance, the elevations so low, far apart and of such irregu- 
lar size (but always small) that its roughness is not at first sight 
apparent. In special locations it has the appearance mentioned when 
describing the hypostome, namely, like a very fine morocco leather. 
Here the elevations are very small, of uniform size, closely placed 
and refractile. 


WEIDMAN—PNEUMONYSSUS FOXI, NOV. SP. 41 


One such special area has already been described over the hypos- 
tome. A second lies ventrally, suggesting a sternal plate. It is 
median and extends from the interval between the first pair of legs 
to a line between the middle of the third pair. It is about three 
times as long as broad, subelliptical, with both ends flattened. It 
is not quite so sharply marked off from the surrounding integument 
as a plate should be. Just within its lateral margins lie six hairs. 
Two are directly at the anterior margin. The second pair is a little 
farther apart and between the second and third coxae. The third 
pair is separated a distance intermediate between the first and sec- 
ond and lies well anterior to the posterior margin of the area, that 
is, about opposite the middles of the third coxae. All hairs are cirected 
posteriorly. 


A third special area lies dorsally, again resembles a plate, is by 
far more extensive than the ventral and well marked off from the 
rest of the integument. It extends from a point immediately behind 
the rostrum to one a short distance behind the fourth coxae. It 
is broadly ellipsoidal except that the posterior end is roundly pointed. 
In its widest part it occupies the middle two fourths of the dorsum. 
It bears five pairs of hairs. The first pair is at the anterior mar- 
gin. The second is closer behind and much closer together. The 
third is close behind the second, but now, again, at the lateral mar- 
gins, and is the farthest apart of any of the five pairs. This brings 
it on a line with the posterior border of the second coxae. The 
interval between the third and fourth pair is about the same as that 
between the first and third. The fourth lies about the same distance 
below the fourth as that between the third and fourth pairs. These 
last two are very close together, being the closest of all the pairs. 
Ail of the hairs lie in the anterior two thirds of the area, the nar- 
rower posterior third being quite naked. This shield bears many 
small groups of pits commonly ascribed to the traction of subja- 
cent muscular attachments. These groups are on the whole of linear 
arrangement, paralleling the scutal border (see plate). 


The fourth special area lies around the anus. This orifice is ter- 
minal and round. In some specimens it is everted and projects beyond 
the general body contour. This is doubtless a pressure artefact. The 
special perianal area is subelliptical, with its longer dimension placed 
longitudinally. Three equidistant hairs lie at its margins, two lateral 
ventral and one median dorsal. They arch over the anal orifice. 
No lateral thorns are seen, although in one specimen the frac- 
ture and eversion of the anal margins by pressure gave this 
appearance. 


42 THE JOURNAL OF PARASITOLOGY 


In addition to the hairs which have been mentioned in connec- 
tion with special areas, a dorsal pair is occasionally found at the 
level of the broadest part of the body posteriorly and far apart, 
lying well lateral to the black lines produced by the intestines. A 
ventral pair is also at times discoverable posterior to the plane of 
greatest body width. These last two pairs are not constant. Those 
of the three special areas are constant. All corporeal hairs are 
inserted in a special ring similar to the ones mentioned in connec- 
tion with the hairs on the legs, and all are directed posteriorly. 


There is but one pair of stigmal plates. They lie between and 
slightly dorsal to the third and fourth coxae and are about three 
times as long as broad, the narrower end directed cephalodorsad, the 
broader end containing the orifice. They show two or three faint 
curved transverse lines in imitation of segmentation. 


The vulvar orifice lies ventrally in the middle of the transverse 
bridge joining the fourth coxae. It is longitudinal, fissural, short, 
and flanked by narrow, linear, chitinous plates. It appears to be 
continuous above with the posterior angle of a laterally elongated 
triangular opening whose other two angles extend far laterally along 
the chitinous bridge. 

The intestines are indicated by two deep, black, tortuous lines 
extending longitudinally close to cuticle dorsally. 


THE MALE 


The solitary male discovered is adjudged such from its slenderer 
proportions (it measures 0.55 X 0.25 mm.), from the presence of a 
special anterior ventral orifice, and the lack of the vulvar orifice. In 
most other respects it is identical in external appearance with the 
female. All legs have the two dorsal hooks and one ventral piece, 
there is no caroncle on the first, short broad ones on the two middle 
ones and a longer, slenderer one on the fourth. The chelicerae in 
this specimen are, by chance, far extended. At most only the tips 
of the fingers happened to project in the case of the females. As 
shown in this male, each is projected from a sheath, extending a 
short distance beyond the hypostome. Each chelicer has a sharply 
pointed, lateral, longer, and median shorter, finger, both springing 
from a common base. The lateral one bends mesially and anteriorly, 
describing a compound curve. The median one curves upward. 


The genital orifice of the male appears close behind the hypos- 
tome as a small circular aperture. From it a tube leads posteriorly 
for some distance directly under the ventral cuticle in a shelving 
manner, so that to superficial inspection it appears like a median 
longitudinal furrow. 


WEIDMAN—PNEUMONYSSUS FOXI, NOV. SP. 43 


THE LARVA 


A larval form was found close to a ruptured female within which 
no ovum could be found, from which it is surmised that it escaped 
from the latter during technical manipulation. The larva measures 
0.550 mm. X 0.280 mm., is oval and has six legs, folded ventrally 
and with long hairs extending, tuft-like, from the distal segments. 
The anal plate is clearly marked and provided with three hairs. 


ZOOLOGICAL POSITION 


The writer places this parasite in the genus Pneumonyssus only 
tentatively and with much unwillingness. In several respects it does 
not agree with the generic and, furthermore, the superfamily diag- 
nosis. In the former there should be no shields, in the latter no hypos- 
tomal armature. Following Banks’ key, however, there is no alter- 
native. It is felt that the time for a rearrangement of these endo- 
parasitic Acarians is at hand, and in the expectation that this will 
be done in the near future it is deemed inadvisable to attempt to 
announce a new genus for this one species. It should be pointed out 
at this time, however, that this is the first time that a male Pneu- 
monyssus has been described, and that the position of its sexual 
orifice places it close to, if not in, the Gamasidae. Indeed Banks 
has already hinted, in a personal communication, that the genus Pneu- 
monyssus may belong more properly to the Gamasidae than to the 
Dermanyssidae, on the basis of certain features noted in the nymph 
of P. simicola. 

As far as known to the writer, this is the fifth species of arachnoid 
described from the air passages of a monkey. 


Pneumonyssus simicola was found by Banks (1904) in the lungs 
of a Javanese monkey (Cyanocephalus sp. ?) dying of opium poison- 
ing in Java. 

Pneumonyssus duttoni was found by Newstead and Todd (1906) 
in the trachea and bronchi of eleven Schmidt’s monkeys (Cercopithe- 
cus schmidti) in the Congo. 


Pneumonyssus griffithi was found by Newstead (1906) in the 
lungs of six rhesus macaques killed in England after exposure to 
tuberculosis. 

Pneumotuber macaci was found by Landois and Hoepke (1914) in 
the lungs of a Macacus rhesus killed in Breslau. 

The description of no one of these four species agrees with this 
parasite or permits its inclusion in that species. 


44 THE JOURNAL OF PARASITOLOGY 


Pneumonyssus simicola has “a broad pulvillus beneath the claws 
in some specimens, probably females,” whereas a pulvillus is con- 
stant on legs i, ii, and iii in P. foxi. P. simicola has four small bris- 
tles on dorsum, P. fori has ten. With P. simicola bristles are not 
present on the coxae. On P. foxi they are commonly present. P. 
sinucola has no dorsal plate as has P. fo.t. 


P. duttoni is at once excluded by the presence of a transverse 
bedy division, two pairs of stigmal plates and its very elongate form. 
It has a dorsal shield. 


P. griffithi has stiliform mandibles; those of P. foai are chelate. 
The dorsal shields of the former has six hairs, those of P. foxi ten. 
The arrangement of the groups of pores (pits) here is different, 
too. 


Pneumotuber macaci has no shields, only one claw on dorsum of 
tarsi i, 1, and@ aiiand a pulvillus on tarsus iv only. There are four 
dorsal hairs as against ten for P. foxi. 

Finally, and of most importance, with none of the above species 
is a ventral plate or the special features of the hypostome mentioned. 
It is true that these are easily overlooked and may have been pres- 
ent in the other species, but until this is shown the parasite here 
described must remain a new species, though much needed future 
rearrangement is likely to place it in a different genus or family. 

The technical description of the new species follows: 


Class, Arachnoidea; order, Acarina; superfamily, Gamasoidea; diagnosis 
(Banks) : Hypostome small, without teeth; venter without furrows; body often 
with coriaceous shields; posterior border never crenulate; no eyes. Family, 
Dermanyssidae; diagnosis (Banks): Parasitic on vertebrates; mandibles fitted 
for piercing; body sometimes constricted. Genus, Pneumonyssus; diagnosis: * 
A Dermanyssid; stigmal plate a little more than twice as long as broad, situ- 
ated above and between the coxae of the third and fourth pairs of legs. Body 
without apparent shields; mouth parts retracted in the head, the palpi very 
short, scarcely visible; the mandibles have apparently both fingers very slender, 
elongate and pointed, probably used for pricking the tissues. The legs are stout 
and short, subequal in length, none as long as width of body, each terminated 
in two subequal claws. Body nearly twice as long as broad, in the male more 
slender. Legs with stiff bristles, but body nearly destitute of hairs. 

Type species, P. simicola Banks. 

P. foxi = n. sp.; diagnosis: Adult females, yellowish white, opaque, in width 
a little more than half the body length. Dorsal shield with ten hairs and 
pitted areas, ventral with six hairs. Anal plate present with three hairs. All 
tarsi furnished with two dorsal claws, all except leg i with caroncle in addition ; 
all articles hirsute and most also spinulose. Both tarsi and femora subdivided. 


* Kindly furnished, together with other information, in a personal communi- 
cation by Dr. Nathan Banks. 

+ Dedicated to Dr. Herbert Fox, who performed the autopsy upon the 
animal, recognized the parasitic nature of the lesions and submitted all the 
material to the writer for identification. 


PLABE- 1 


OGO 


}3 


WEIDMAN—PNEUMONYSSUS FOXI, NOV. SP. 45 


Palpi of three segments, all short, the terminal one capped by a short bristle. 
Mandibles chelate in both sexes. One pair of stigmal plates between and dorsal 
to coxae iii and iv. Hypostome bears a median longitudinal row of 9 to 13 
teeth, carries ten hairs anterolaterally and four anterior marginal papillae. 
Vulva short, median and fissural at level of coxae iv. Adult males measure 
a little less than half as wide as long. Sexual orifice circular and close behind 
capitulum. Larva hexapod, oval, 0.55 0.28 mm., bears anal plate. Length 
0.875 mm., breadh 0.478 mm. 

Fabist, lungs of monkey (Macacus rhesus). 

Autopsy number P. Z. G. 3156. 


ARTICLES CITED 


Banks, N. 1904. A Treatise on the Acarina, or Mites. Proc. U. S. Nat. 
Mus., 28: 1-114. ; 

Haan, J. de. 1906. Gibt es beim Menschen endoparasitar lebende Acariden? 
Centralbl. f. Bakteriol., Abt. 1, Orig. 40: 693-4. 

Landois, F., and Hoepke, H. 1914. Eine endoparasitare Milbe in der Lunge 
von Macacus rhesus: Centralbl. f. Bakteriol., Abt. 1, Orig. 73: 384-91. 

Newstead, R. 1906. Another New Dermanyssid Acarid. Liverpool School 
Trop. Med., Mem. 18. 

Newstead, R., and Todd, J. L. 1906. A New Dermanyssid Acarid Found 
Living in the Lungs of Monkeys (Cercopithecus schmidti) from the Upper 
Congo. Report of the Exp. to the Congo, 1903-05. Liverpool School Trop. 
Med., Mem. 18. 


EXPLANATION OF PLATE 

Fig. 1—Ovigerous female viewed ventrally. Magnification about 66 times. 

Fig. 2—Adult (?) male viewed ventrally. Magnification about 66 times. 

Fig. 3—Same as Figure 2 but more highly magnified. Shows mouth parts 
and ventral shield, the latter with genital orifice and six hairs. Magnification 
about 240 times. 

Fig. 4—Teased adult female: ds, dorsal shield; vs, ventral shield; mf, 
muscle fasciculi. Magnification about 100 times. 


Fig. 5—Dorsal shield from Figure 4 more highly magnified; pp, pits; mf, 
muscle fasciculi. Magnification about 260 times. 
Fig. 6.—Leg ii and ili: ca, caroncle; ta, tarsus; p, patella; ti, tibia;-f, femur; 
tr, trochanter; co, coxa. 
Fig. 7—Extremity of Leg 1. 
Fig. 8—Extremity of Leg iv. 
Fig. 9.—Stigmal plates. 
Fig. 10.—Dorsal shield. Compare with Figure 5. 


CESTODE CYSTS FROM MUSKRAT 


Epwin LINTON 
Biological Laboratory, Washington and Jefferson College 


The material was collected from a muskrat found near Wash- 
ington, Pa., on Feb. 8, 1884. Four cysts were found, three embedded 
in the liver, and one in the peritoneum. The cysts were elliptical in 
outline, the largest measuring 13 by 9 mm. When opened, the con- 
tained cysticercus was seen to have developed into a strobila, with 
the bladder portion reduced to a small, flattened, spatulate body with 
collapsed walls. 

The strobiles were milk-white and actively contractile. When first 
released they showed a tendency to thicken anteriorly so that the 
whole strobila became more or less clavate. Two of them, measured 
immediately after removal from the cysts, were 121 and 143 mm. 
respectively. After lying in water over night the larger specimen 
measured 212 mm. A few hours later it measured 300 mm. The 


Fig. 1—Pair of hooks. Length of longer hook 0.4 mm. 


anterior end, for a distance of about 175 mm. was about 6 mm. in 
breadth and 3 mm. in thickness. The posterior third narrowed uni- 
formly to 2 mm. The remnant of the bladder at the posterior end 
was 6.5 mm. in length and 5 mm. in breadth. When this specimen 
was placed in alcohol at the end of forty-eight hours it measured 
325 mm. 


The diameter of the scolex in a mounted specimen is 1.06 mm. 
The suckers are rather prominent and directed anteriorly. The por- 
tion of the scolex in front of the suckers, when the hooks are com- 
pletely everted, is conical-truncate. There are two circles of hooks, 
those in the anterior circle being the larger. The hooks lie in 
pairs, a pair consisting of a large and a small hook (Fig. 1). The 
number of hooks, estimated from a study of living specimens, seen 
lateral view, was fourteen in each circle. Another specimen seen in 


LINTON—CESTODE CYSTS FROM MUSKRAT 47 


front view had eighteen hooks in each circle. The hooks of the anterior 
circle are about 0.45, andythose of the posterior circle 0.26 mm. long. 

Proglottids begin a very short distance back of the suckers where 
they are about 0.5 mm. in length. In the larger alcoholic specimen 
the proglottids toward the middle of the length are 5 mm. in 
breadth and 0.72 mm. in length; farther back the breadth is 4 mm. 
and the length 0.8 mm.; toward the posterior end the breadth is 
3 mm. and the length 0.56 mm. Sinuous marginal vessels are visible 
in the stained and mounted segments, but no rudiments of genitalia 
were seen. 

The size and shape of the hooks and the appearance of the bladder- 
worm indicate that this cestode is the form known as Cysticercus 
jasciolaris the larval stage of Taenia crassicollis. 


SARGOPHAGID LARVAE FROM THE PAINTED 
TURTLE 


F. E. CHIDESTER 
Rutgers College, New Brunswick, N. J. 


In studying the blood of vertebrates in my course in histology dur- 
ing the past winter, I used a specimen of Chrysemys picta, the common 
painted turtle. In pulling the right hind leg back against the plastron 
before making the stab for blood, I noted that a hardened cylinder 
partly protruded from the thigh. When the object was completely 
extruded, it parted in the middle and five sarcophagid larvae were 
discovered. 


Figure 1 Figure 2 


Fig. 1—Horny case containing Sarcophagids. 
Fig. 2.—Sarcophagid larva from the turtle. (Ventral aspect.) 


The specimens were submitted to Dr. T. J. Headlee and to Dr. L. 
©. Howard. For information regarding the larvae, I am indebted 
to them and to Mr. C. H. Richardson, assistant state entomologist of 
New Jersey. 

So-called bot-fly larvae have been reported by Packard (1882), 
True (1884), and Wheeler (1890). Wheeler corrected the previous 
writers and placed the larvae among the Sarcophagae. Most recently 
Kepner (1912) has described in detail larvae undoubtedly of the same 
species as my own specimens. There is no question that the tortoises 
are infested by sarcophagids and the reason for this brief note is found 
in the peculiar modification of the epidermis of the host which formed 
a case for the larvae. 

This horny epidermal case (Fig. 1) was 15 mm. long, 5.5 mm. 
broad, and from 0.5 to 0.6 mm. thick. The outside was regular and 
smooth except at one point, where it was slightly indented. The 


CHIDESTER—SARCOPHAGID LARVAE 49 


mouth of the case was irregular and before being disturbed the object 
was scarcely discernible on the skin of the turtle. The hind leg 
was measured, and the thigh proved to be only 25 mm. in length and 
13 mm. in diameter. The leg was completely paralyzed, and although 
the specimen was kept alive for a month after the case had been 
removed, use of the leg was not regained. 

The larvae (Fig. 2) were kept alive for some days, then two were 
placed in moist earth in an attempt to secure pupation. The results 
were negative, however. The other specimens were turned over to 
the entomology department and one of them was sent to the depart- 
ment of agriculture for identification. So far as the writer knows 
no imagoes other than the four females secured by Wheeler have been 
bred from the turtle-infesting sarcophagid. 


LITERATURE CITED 


Kepner, W. A. 1912. The Larva of Sarcophaga, a Parasite of Cistudo Caro- 
lina and the Histology of Its Respiratory Apparatus. Biol. Bull., 22: 163-172. 

Packard, A. S. 1882. Bot-Fly Larvae in a Turtle’s Neck. Am. Nat., 16: 598, 

True, F. W. 1884. Bot-Flies in a Turtle. Science, 4: 511. 

Wheeler, W. M.: 18°90. The Supposed Bot-Fly Parasite of the “Box-Turtle”. 
Psyche, 5: 403. 


NOTES 


The life-history of the human blood fluke is undoubtedly one of the most 
important of unsolved problems in parasitology. For many years, in fact ever 
since the discovery of the adult parasite in Egypt by Bilharz in 1852, this ques- 
tion has commanded the attention of able investigators, but practically no posi- 
tive results have followed their work. Local tradition supported by circum- 
stantial evidence ied to general acceptance of the view that infection with the 
African species, Schistosoma haematobium, was acquired by bathing in infected 
waters. After extended study in regions of pronounced infection Looss con- 
cluded that infection took place directly through the skin and that the infecting 
stage was the miracidium, which underwent metamorphosis in the body of the 
final host, probably in the liver. 

The discovery of a new human species (Schistosoma japonicum), was aug- 
mented by its later demonstrated occurrence in various small mammals and 
experimentation became possible. Utilizing this opportunity, Leiper and Atkin- 
son recently made a trip to the East and as a result of their work, which was 
unfortunately cut short by the war, have published * most important studies 
on the life history of this fluke. 

After a search lasting nearly three months and a river journey of a thousand 
miles, they secured a dog so heavily infected that the evacuations consisted of 
mucus and blood crowded with eggs. The local mollusks were placed in water 
swarming with miracidia and were watched to detect those species which exer- 
cised a pronounced attraction for the free-swimming fluke embryos. A _ small 
brown snail of a new genus, Katayama nosophora, displayed such an attraction, 
“The small dark head and foot speedily became festooned with little white 
specks and it was obvious from the agitated manner in which the snail repeatedly 
attempted to brush them off that their presence was a cause of considerable 
irritation.” 

In the liver of this snail were sporocysts containing cercariae with bifid tails. 
The cercaria had a short bifurcated gut with no trace of a pharynx. Labor- 
atory-bred mice were exposed to infection in water containing free cercariae 
from the. teased snail liver. At Aden on the home voyage the few mollusks 
living were sacrificed, and the last mouse was exposed to infection. When 
examined in London a month later this mouse contained live male and female 
blood flukes in copula in the portal vessels. These factors show conclusively 
that this schistosome has a life-cycle like that of the digenetic trematodes. 

The cercaria is covered by minute spines, the oral sucker is enormous, equal 
to about one-third the length of the body, and urn shaped. Between the lateral 
branches of the intestine are several masses on each side, the undeveloped sex 
glands. The snail which functioned as secondary host though abundant proved 
to be entirely new. 


* Brit. Med. Jour., January, 1915; China Med. Jour., May, 1915. 


PROFESSOR STANISLAUS VON PROWAZEK 


The Journal of Parasitology 


Volume 2 DECEMBER, 1915 Number 2 


PROFESSOR von PROWAZEK * 
(WITH PORTRAIT) 


Im Dienste der Wissenschaft und des Vaterlandes starb am 17. 
Februar Professor Stanislaus von Prowazek, der Leiter des Protozoen- 
laboratoriums am Institut fiir Schiffs- und Tropenkrankheiten in 
Cottbus an Flecktyphus. In ihm verliert nicht nur das Tropeninstitut 
sein bedeutendstes Mitglied, sondern die gesamte wissenschaftliche 
Welt einen Forscher von universeller Bedeutung. 

Prof. Dr. v. Prowazek war 1875 in Oesterreich geboren. Er stu- 
dierte in Prag und Wien. Nachdem er kurze Zeit Assistent am 
Institut fiir experimentelle Therapie in Frankfurt a.M. (Direktor Geh. 
Rat Ehrlich) und am zoologischen Institut der Universitat Miinchen 
(Prof. Hertwig) gewesen war, wurde er 1903 auf Veranlassung von 
Fritz Schaudinn, dem damaligen Vorsteher der Abteilung fiir Proto- 
zoenforschung des Kaiserlichen Gesundheitsamtes dorthin nach Berlin 
berufen und wurde, als Schaudinn dem Rufe ans tropenhygienische 
Institut zu Hamburg folgte, der provisorische Leiter des Protozoen- 
laboratoriums im Kaiserlichen Gesundheitsamt und nach dem allzu 
frihen Tod des genialen Schaudinn sein Nachfolger in Hamburg 
(1907). Er war mit Schaudinn aufs innigste befreundet und hat nicht 
nur die weitausblickenden Gedanken dieses Forschers nach seinem 
Tode aufs gliicklichste weiter verfolgt, sondern auch die Wissenschaft 
mit vielen, glanzenden, selbstandigen Gedanken und Beobachtungen 
bereichert. Seine Studien tiber die Physiologie und Biologie der Zelle 
und der Protozoen im besonderen, seine Untersuchungen tiber Variola 
und Vakzine, tiber Trachom, Blennorrhoe und andere Augenkrank- 
heiten und die darauf gegriindete Erfassung der als Infektionserreger 
weit verbreiteten Chlamydozoengruppe (Pocken, Hundswut, verschie- 
dene Augenkrankheiten und Tropenkrankheiten) machten ihn zur 
bedeutendsten Autoritat unter den Forschern auf dem Gebiete der 
modernen Protistenkunde. 

Kurz nach dem Tode Schaudinns hatte er die Ausreise nach Niedey. 


*This sketch was written for THe JouRNAL on request by Prof. W. 
Michaelsen of the Naturhistorisches Museum in Hamburg. 


52 THE JOURNAL OF PARASITQLOGY 


landisch-Indien als Mitglied der Neisser’schen Syphilis-Expedition 
angetreten, gelegentlich derer er mit Halberstadter seine Entdeckungen 
iiber die Aetiologie des Trachoms machen konnte; nach seiner Riick- 
kehr trat er die Stelle am Tropeninstitut am 16, Juni 1907 an. 

Hier am Institut entfaltete er eine uberaus fruchtbare Tatigkeit, 
teils in stiller Laboratoriumsarbeit, teils auf ausgedehnten Forschungs- 
reisen. Von diesen fiihrte ihn eine vom 10. Juni 1908 bis 26. Februar 
1909 an das Instituto Oswaldo Cruz nach Rio de Janeiro, wohin er 
gemeinsam mit Prof. Giemsa auf Einladung des brasilianischen Staates 
beurlaubt wurde zu Lehr- und Forschungszwecken. Von Mitte 1910 
bis Ende 1912 machte er gemeinsam mit dem Ophthalmologen Dr. 
Leber mit Mitteln des Reichskolonialamts und des Hamburgischen 
Staates eine Expedition nach Sumatra und dem deutschen Siidsee- 
gebiet zum Studium der Granulose und anderer Krankheiten. Die 
Frichte dieser Expedition waren nicht nur auf seinem Spezialgebiet 
reichlich; sondern er hat auch in seinem Buche “Die deutschen 
Marianen” dank seinem universellen Wissen die Geschichte, Ethno- 
graphie, Fauna, Flora und das Medizinische in tiberaus gewissenhafter 
und poetisch-reizvoller Form veroffentlicht. Spatere Reisen fthrten 
ihn zum Studium des Flecktyphus im Sommer 1913 gemeinsam mit 
Hegler nach Serbien und im Sommer 1914 nach Konstantinopel mit 
Rocha-Lima. Mit letzterem wurde er, als im Dezember 1914 im Rus- 
senlager in Cottbus eine grosse Fleckfieberepidemie ausbrach, vom 
Kriegsministerium mit wissenschaftlichen Untersuchungen daselbst 
betraut. Dort ist er mn der ersten Februarwoche 1915 an der Seuche 
erkrankt und der Infektion am 17. Februar erlegen. 

Prowazek war ein Forscher von ungewohnlich vielseitigem Wissen ; 
er beherrschte nicht nur meisterhaft das Gebiet der Zoologie und ihre 
Grenzgebiete, sondern besass auch auf dem Gebiete der Botanik, 
Physik, Chemie, und Philosophie ungemeine Kenntnisse, ebenso auch 
auf medizinischem Gebiet, besonders der Immunitatslehre. Dies zeigt 
sich in seinen zahlreichen Arbeiten auf diesen Gebieten ausgepragt. 
Prowazek war ungeheuer fleissig und seine Arbeiten zeichnen sich 
durch eine erschopfende Griindlichkeit aus. Fur den Mediziner sind 
vor allem bedeutungsvoll geworden seine Studien tiber die Biologie und 
Physiologie der Zelle und der Protozoen im speziellen. Sein Buch 
“Die Physiologie der Einzelligen (Protisten)” enhalt eine Fille von 
ihm gefundener neuer Tatsachen und die Erorterung neuer Probleme. 
Vor allem die Tropenmedizin verdankt Prowazek als Lehrer und 
Forscher sehr viel; viele unserer Tropenarzte, die ihm als Schiller und 
Freunde naher getreten waren, standen dauernd mit ihm in regem 
wissenschaftlichem Gedankenaustausch. 


PROFESSOR VON PROWAZEK 53 


Am Tropeninstitut hat er auch eine ausgedehnte Lehrtatigkeit ent- 
wickelt und Schiiler aus allem Welt fesselte er an sich. Mit Schiilern 
und Freunden als Mitarbeitern hat er auch das gross angelegte “Hand- 
buch der pathogenen Protozoen” herausgegeben, dessen Abschluss- 
band, fur den von ihm selbst zahlreiche fertige Manuskripte vorliegen, 
er nicht mehr erleben konnte. Das von Schaudinn begriindete “Archiv 
fur Protistenkunde” hat er gemeinsam mit Hartmann weitergefiihrt 
und zur ersten grossten, internationalen Zeitschrift auf diesem Gebiete 
gestaltet. 

Prowazek war ein Mensch, der wenig gern in die Oeffentlichkeit 
trat, still und zuriickgezogen lebte, eine echte Gelehrtennatur. Wer 
aber das Gliick hatte, ihm naher treten zu diirfen, lernte ihn als Men- 
schen von seltenem Wissen und feinster Kultur kennen, begabt mit viel 
Sinn fur alles Schone in Kunst und Natur. Im Kreise seiner Tatigkeit 
war er verehrt und geschatzt von allen, bot stets eine Fille von 
Anregungen. So ist sein Tod nicht nur ein unersetzlicher Verlust fiir 
die Wissenschaft, sondern auch ftir seine vielen Freunde. 


FURTHER NOTE UPON COMPARISON OF ENDAMOEBA 
GINGIVALIS (GROS) AND ENDAMOEBA 
WISTOLYTICA SCHAUDINN 


ALLEN J. SmitH AND M. T. BarreETT 


From the picManes Pathological Laboratories, School of Medicine, 
University of Pennsylvania 


In a recent number of this Journal the writers, after an analysis 
of the records of discovery of parasitic amebae in the human mouth 
concluded that the oral endamebae of man are referable to two species: 
Endamoeba gingivalis (Gros 1849) and Endamoeba pyogenes (Verdun 
and Bruyant 1907). In an attempt to compare these oral parasites with 
other parasitic amebae of man the writers suggested that the second 
species named may be identical with an organism, characterized like it 
by a large nucleus containing a large, granular, richly chromatinized 
binnenkérper, found by Ribbert in the ducts of the parotid gland, and 
believed by the writers to be the same as found some years ago by Rib- 
bert in the renal tubules of a syphilitic new-born infant, by Jessionek 
and Kiolemengolou in the kidneys, liver, and lungs of an aborted syph- 
ilitic fetus, and by Smith and Weidman in the kidneys, lungs, and liver 
of a non-syphilitic new-born infant and in the lungs of a syphilitic infant 
one month old, to which these last writers have applied the name 
of Endamoeba mortinatalium. (For literature cf. original article in 
this Journal.) In comparing the first named species of oral endamebae 
with Endamoeba histolytica Schaudinn, the writers asserted so close a 
morphological similarity that, while unwilling to declare the biological 
identity of these parasites, they felt unable by microscopic examination 
alone to differentiate between them. Emphasis was laid in a footnote, 
added to the paper after its presentation at the Christmas convocation 
of 1914 before the Association of American Bacteriologists and for- 
warded with the copy for publication, that this statement had reference 
to Endamoeba histolytica solely in its histolytica phase (not the tetra- 
gena phase) ; and the same footnote contained a brief record of failures 
of feeding experiments, thus adding reason for believing in the duality 
of the species altogether apart from the morphological similarity pre- 
sented. Unfortunately this footnote was omitted in printing; and 
through oversight an editorial note, written following a discussion by 
correspondence, was inserted, emphasizing the tetragena phase of 
Endamoeba histolytica as a basis of differentiation and urging the known 


1. Jour. of Parasitol., June, 1915, vol. 1, pp. 159-174. 


aid Ss 


COMPARISON OF ENDAMOEBAE 55 


reproductive encystment in this latter phase as a point in separation. 
The writers have felt thatethereby their position has been the more 
opened to misconception, and through the kindness of the editor are 
publishing the present note to amend and define their original state- 
ment, and at the same time to record briefly their attempts to induce 
colonic infestment by oral endamebae. 

In the histolytica stage of Endamoeba histolytica Schaudinn the 
parasite is known to divide by fission and, many at least believe, also 
by gemmation, but there is no reproductive encystment. In the tet- 
ragena stage of the same parasite the true encystment with four off- 
spring occurs. In the histolytica stage the nucleus is practically 
invisible in the unstained state ; in the tetragena stage it becomes visible 
unstained, and when stained shows a thicker nuclear membrane, a 
larger binnenk6érper and a higher chromatinization. Endamoeba gingi- 
valis simulates the first of these phases in its nuclear characters and in 
its apparent modes of reproduction. It is of course to be expected that 
sometime and somewhere reproductive encystment does take place ; and 
it is not impossible that in some other situation than in the gums a phase 
is assumed comparable to the tetragena phase of the dysenteric organ- 
ism, in which reproduction in encystment occurs. We say “elsewhere 
than in the gums” because we have not noted examples suggesting such 
change in the pyorrhea material from the many individuals whose para- 
sites we have seen; and a large proportion of these cases was decidedly 
chronic (the element of chronicity apparently being important to the 
assumption of tetragena characteristics and to encystment reproduction 
in case of Endamoeba histolytica). As far as the question of gemmation 
is concerned doubt of course is natural. But the writers are not satis- 
fied that the separation of gemmules is merely a phenomenon of 
degeneration. We have repeatedly seen the throwing off of gemmules 
by actively moving and apparently normal amebae, and especially in our 
attempts to cultivate the oral endamebae in vitro have found what we 
believed to be such gemmules in motion, and in stained preparations of 
the same material similar small protoplasmic bodies containing a minute 
bit of chromatin. We cannot, of course, declare the fate of these 
separated particles, but the appearances observed certainly make us 
unwilling to accept unhesitatingly the view of their inability to grow 
into adult amebae. 

From the morphological similarities of Endamoeba gingivalis and 
the histolytica phase of Endamoeba histolytica and the occurrence of 
binary fission and gemmation as modes of reproduction of both (with 
no reference to the tetragena phase of the latter and its known mode of 
encystment reproduction), the writers held that methods of differentia- 
tion other than by comparison of morphological features are essential 
for differentiation. Since the presentation of the original paper we have 


56 THE JOURNAL OF PARASITOLOGY 


accumulated negative evidence indicating the duality of these oral enda- 
mebae and the dysenteric parasites, in the constant failure of attempts 
to infest the colon with pyorrhea material rich in Endamoeba gingi- 
valis Gros. In these experiments, in which we were joined by Dr. 
Baldwin H. Liicke, assistant instructor of pathology in this laboratory, 
pyorrhea material bearing active vegetative endamebae was given to 
two kittens by feeding, to two puppies and two kittens by high rectal 
enemata, and to four kittens by injection into the colon after lapa- 
rotomy. In all cases we failed to find amebae in the dejections and to 
note any evidence of dysenteric symptoms, and in all but two kittens to 
meet at autopsy with lesions of the colon at all suggestive of success. 
In these last a few ulcers were met, but smears made from the surface 
of the ulcers, and serial sections of the lesions, failed to show the pres- 
ence of amebae. Such failures are not infrequent, it is true, when 
material known to contain Endamoeba histolytica is employed; and a 
single positive result would outweigh the negative results of our 
attempts. But because of the uniformity of failure of our experiments 
we feel that the original impression of biological difference between the 
species must be maintained in spite of the morphological similarities 
presented. With such a belief we are disposed to say in spite of the 
possibility of complete morphological similarity of individual examples 
of Endamoeba gingivalis (Gros) on the one hand and of the histolytica 
type of Endamoeba histolytica Schaudinn on the other, that one should 
in general find that individuals of gingivalis are slightly smaller, some- 
what less active, with pseudopods commonly more lobose, with a nucleus 
more frequently central in position, and with a more actively hemolytic 
capacity (more rapidly destroying englobed erythrocytes and therefore 
ordinarily showing fewer red cells in the body of the parasite) than 
will be the case with individuals of Endamoeba histolytica in the his- 
tolytica phase and that intestinal infestment by gingivalis is probably 
impossible. We accept without hesitation the existence of definite dif- 
ferentiating morphological features to separate the oral parasite from 
the tetragena phase of Endamoeba Mstolytica. 


. 


NOTES ON THE TREMATODE GENUS TELORCHIS 
WITH -DESCRIPTIONS .OF NEW. SPECIES* 


Horace W. STUNKARD 


In 1889 Liihe created a new genus, Telorchis, to contain certain rep- 
tilian distome parasites, and designated D. clava Diesing (1850) as the 
type species. In the genus he included D. poiriert Stoss. (=D. 
gelatinosum Poirier nec. Rud.), D. linstowit Stoss. (== Monostomum 
aculeatum v. Linst.), D. ercolanu Montic. (= D. signatum Ercol. nec. 
Duj.), D. nematoides Muhl., D. bifurcum Braun, D. pleroticum Braun, 
and tentatively D. arrectum Mol. nec. Duj. His characterization of the 
genus states that the testes lie behind one another at the posterior end 
of the body; the cirrus sac opens somewhat left of the acetabulum and 
is very long; the ovary is immediately behind the posterior end of the 
cirrus sac and is*separated from the testes by the coils of the well 
developed uterus; while the vitellaria consist of numerous follicles 
occupying the space at the sides of the body and approaching more or 
less closely the anterior and posterior ends. The diverticula of the 
intestine reach almost to the posterior end of the body; and with the 
exception of D. poirieri all species are armed with spines at the cephalic 
extremity of the worm. The excretory vessel is long and branches 
anteriorly in the form of a Y. The oral sucker is usually slightly larger 
than the acetabulum, though in D. ercolanii of the same size. 

This same group of reptile distomes was separated by Looss (1899) 
independently, and also called Telorchis, but his article appeared after 
that of Lithe. Looss selected D. linstowi as the type species. 

Because of the differences existing between 7. clava and the other 
members of the genus, Lihe later (1900) created two subgenera: 
Telorchis with T. clava as type, and Cercorchis with T. aculeatus (= T. 
linstowt Stoss.) as the type. The distinguishing features of the sub- 
genus Telorchis are stated as the absence of an esophagus and the 
lateral extension of the folds of the uterus beyond the diverticula of 
the intestine where they may be coiled over the ceca in the form of a 
figure 8. In the sub-genus Cercorchis an esophagus is present and the 
coils of the uterus are confined between the ceca. 

My examination of almost a hundred mature individuals of six 
different species affords evidence that the lateral extension of the uterus 
varies largely as a result of congestion with eggs. In the same species 
one finds some specimens in which the coils of the uterus are confined 


* Contributions from the Zoological Laboratory of the University of Illi- 
nois, under the direction of Henry B. Ward, No. 55. 


58 THE JOURNAL OF PARASITOLOGY 


between the ceca and others in which the uterine folds overlap the 
diverticula on one or both sides. Certain species in the genus possess 
a long esophagus, others a short esophagus, and finally in T. clava an 
esophagus is absent. Furthermore the absence of an esophagus is not 
always associated with an extracecal coiling of the uterus, and vice 
versa, since in T. bifurcus an esophagus is absent and the uterine coils 
are intracecal, and in T. corti an esophagus 1s present and the uterine 
folds often overlap the ceca. These facts show that the characters 
designated by Lithe are not adequate to subdivide the genus, and since 
the apparent morphological differences of his types are merely extreme 
variations of characters common to several species, the sub-genera 
disappear. 

Goldberger (1911) described as new species T. stossichi, T. attenu- 
atus and T. robustus, and formulated a key for the identification and 
separation of the species. 

The genus has a wide distribution, species having been reported 
from Sicily, Sardinia, Italy, France, Germany,  Austro-Hungary, 
Turkey, Brazil, United States and Canada. So far as is known it is 
confined to reptilian hosts, species occurring in lizards, snakes and 
turtles. 

The trematodes in this genus are elongate, with more or less parallel 
sides. The region of greatest width is at or anterior to the middle of 
the body. They range in length from 1.5 to 13 mm. and in width from 
0.25 to 16mm. In T. arrectus the ratio of width to length is 1:4, in 
T. diminutus it is 1:5, in T. clava, T. bifurcus and T. lobosus it is 
about 1:7, while in T. pleroticus it is 1:18 and in T. poirieri it is 1: 22. 
That part of the body between the oral sucker and the acetabulum is 
much more motile than the post acetabular region, which is essentially 
a sac containing the reproductive apparatus. 

The cuticula is of uniform thickness in any one worm; it varies 
from 2p in T. parvus and T. diminutus to 11p in T. attenuatus, the 
thicker cuticula being found in the larger species. Cuticular spines 
occur on the body arranged in a quincunx pattern and around the 
external openings in concentric circles. They are deeply imbedded in 
the cuticula which is raised about the base of each spine in a papilla 
like structure. Largest around the oral sucker, they gradually diminish 
in size toward the posterior end of the body where they are indistinct 
or entirely absent. The rows are separated by distances slightly exceed- 
ing the length of the spines. In general, the spines vary in size and 
proximity directly with the size of the worm. In one specimen of T. 
robustus 13 mm. long, the spines near the anterior end are 6.5 apart 
and 5.7 in length, and in another 6 mm. long they are 3.2 apart and 
3 in length. In specimens of T. lobosus they are 2u long and in rows 


NOTES ON THE TREMATODE GENUS TELORCHIS 59 


2.2u apart, and in T. diminutus they are 1.54 long and in rows 
1.6. apart. The cuticulagturned in at the external openings is not 
spinous. 

The musculature of the body (Fig. A) is light and delicate so that 
the worms are translucent. 

The excretory system is typical for the genus. The pore is situated 
at the posterior tip of the body and opens from a large, median collect- 
ing duct (Fig. B) which extends anteriorly to about the location of 
the ovary; there it divides into branches that extend anteriad, one on 
either side of the median line, just mesal to the intestinal diverticula. 
These branches can be traced almost to the region of the acetabulum 
where they disappear. In studying living specimens of T. corti I have 
been able to distinguish the flame cells, but their ducts could not be 
followed. 


~ ses 


PERF ----- 1m 


Fig. A Fig. B. 


Fig. A. Tangential section of body wall in T. lobosus, showing muscle 
layers and cuticular spines; cu, cuticula; cm, circular muscles; /m, longitudinal 
muscles; om, oblique muscles. 

Fig. B. Sagittal section at median posterior end of body in T. robustus; 
ep, excretory pore; ex, collecting duct of excretory system; ¢, caudal testis. 


The oral sucker is sub-terminal in position, equalling or slightly 
exceeding the acetabulum in size. The shape of both the oral sucker 
and acetabulum is subject to considerable variation as can be observed 
by watching the movements of a living worm. The shape of the sucker 
at the time of killing and the character of the reagents used influence 
the shape of the organ in the fixed material, although there seems to 
be a certain relation between the general shape of the sucker and the 
species. ; 

A prepharynx is absent in T. aculeatus and T. nematoides and in 
some forms can be noted only when the cephalic extremity of the worm 
is much extended. All degrees of variation in length occur to a distinct 


60 THE JOURNAL OF PARASITOLOGY 


and elongated pouch in T. bifurcus and T. pleroticus. The pharynx is 
approximately spherical although either the longitudinal or transverse 
diameter may be greater. An esophagus is absent in T. clava and in 
T. pleroticus, short in T. aculeatus and T. parvus, and very long in 
T. medius and T. solivagus. The ceca meet anteriorly at an acute 
angle and extend almost to the posterior end of the body, terminating 
rarely (TI. parvus and T. poirieri) in the inter-testicular zone, and in 
all other known species behind the caudal testis. Anterior to its bifur- 
cation the digestive tract is lined with cuticula continuous with that 
of the external surface, and the ceca are lined with digestive epi- 
thelium, cells with nuclei close to the fibromuscular wall and cyto- 
plasmic processes extending into the lumen of the canal. If the contents 
of the ceca are forced caudad the ends may be bulbous or flasklike in 
appearance, while if the caudal part of the excretory duct 1s much 
distended the ceca must necessarily taper gradually. 

The testes lie in close proximity, one behind the other near the 
posterior end of the body, in the median line or slightly to the right 
and left. In his description of T. parvus, Braun (1901) states that due 
to the flattened condition of the body the collecting duct of the excre- 
tory system passes between the testes in the shape of a letter S so 
that when it is distended it causes the testes to lie obliquely. In most 
cases the excretory duct is dorsal to the testes and conditions are not 
those in T. parvus. The vasa efferentia pass cephalad, just median to 
the ceca, the duct from the cephalic testis on the left and that from 
the caudal testis on the right. The ducts move mediad and dorsad as 
they pass forward; at the region of the ovary they pass above the 
excretory tubes and then on the median side of these tubes to the 
posterior end of the cirrus sac where they empty into the vesicula 
seminalis. The cirrus sac (Fig. 1) is a long, cylindrical, muscular 
pouch extending caudad from the genital pore and enclosing the cirrus, 
vas deferens, prostate, and seminal vesicle. The genital pore is immedi- 
ately anterior and at the left of the acetabulum. In one specimen of 
T. corti the cephalic testis had divided, the resulting organs lying one 
on either side of the median line. This specimen was not sexually 
mature and the vasa efferentia could not be traced. 

The ovary is in or near the median line, posterior to the acetabulum, 
and usually just anterior to the center of the body. The oviduct arises 
from the dorsal posterior margin and after one or two slight irregulari- 
ties it enlarges to form the ootype which is surrounded by the large 
unicellular glands of the shell gland. Figure 7 shows the structures of 
the female genital system in the vicinity of the ovary. Laurer’s canal 
branches from the dorsal posterior part of the ootype and opens on 
the dorsal surface; the common vitelline duct enters the ootype just 


NOTES ON THE TREMATODE GENUS TELORCHIS 61 


ventral to and at the left of the origin of Laurer’s canal. In some 
species the proximal end of Laurer’s canal is enlarged to form a seminal 
receptacle and in T. aculeatus the enlargement may comprise the entire 
tube which in one case was filled with chromatin granules, in other 
instances no enlargement is present, as in 7. robustus. The uterine 
tube turns first ventrad and then begins a series of irregular, sinuous 
convolutions, extending posteriad to the cephalic testis and returning 
anteriad to the nearly straight metraterm which leads to the common 
genital sinus. The opening of the metraterm is anterior and at the left 
of the opening to the cirrus sac. The descending and ascending coils 
of the uterus occupy separate distinct fields in T. aculeatus and T. 
parvus; in T. bifurcus they overlap and are in some cases indistinct, 
while in T. diminutus and T. nematoides they are so superimposed and 
confused that only rarely can distinct fields be discerned. In T. soli- 
vagus Odhner (1902) reports that the descending and ascending limbs 
of the uterus cross each other to forma figure 8 and other authors men- 
tion this crossing or absence of crossing as a specific character, but in 
T. corti both conditions exist. 

The vitellaria lie laterad of the ceca, and consist of a large number 
of follicles, usually arranged in lobes. Typically there are nine lobes 
on the right and twelve lobes on the left side of the body, although 
there is considerable variation from this condition. There is a tendency 
for the lobes to fuse, reducing the number, and the vitellaria of the left 
side extend farther cephalad and caudad than those of the right. In T. 
aculeatus the lobes are distinct, in most of the species they can be dis- 
tinguished, while in others (T. diminutus and T. robustus) the vitelline 
follicles are not separated into lobes but extend along the sides in an 
unbroken series. Longitudinal collecting ducts occupy the median face 
of the vitellaria, and in the region of the ootype short ducts leading 
mediad from these unite near the median line of the body to form the 
vitelline receptacle from which the common yolk duct leads to the 
ootype. 

In all sexually mature worms the uterus contains enormous numbers 
of eggs. They vary in size from 10 by 20, in T. pleroticus to 22.8 by 
40 in T. parvus, and 21 by 41p in T. diminutus. It is interesting to 
note that in the smallest species, T. parvus and T. diminutus, the eggs 
are larger than in the largest species, T. poirieri and T. robustus, where 
they measure only 14 by 23u and 15 by 29n, respectively. 

If the description given by Stafford (1900) and (1905) of T. 
angustus, and that by Barker and Covey (1911) of 7. leptus are con- 
firmed by further study, then these species do not belong in the genus 
Telorchts as conceived and discussed in preceding section of this paper. 
The long distance separating the acetabulum and the genital pore, the 
dorsolateral location of the latter, and the pre-acetabular position of 


62 THE JOURNAL OF PARASITOLOGY 


the cirrus sac form a complex of such striking and fundamental dif- 
ferences that a natural grouping will remove these forms from the 
genus Telorchis, thereby raising the sub-genus Protenes Barker and 
Covey to generic rank. Protenes leptus designated by Barker and 
Covey must be taken as type. 

To Professor Henry B. Ward, under whose direction this work was 
done, I wish to express my appreciation for criticisms and suggestions. 


Telorchis corti sp. nov. (Figs. 1, 4) 


Specimens 4 to 7.15 mm. long; 0.35 to 0.5 mm. wide; greatest width at 
acetabulum, which is 0.14 mm. in diameter, one sixth to one seventh of body 
length from anterior end. Oral sucker same size as acetabulum; cuticular 
spines around former 3.4“ in length. Pharynx 70 to 80# in diameter. Esopha- 
gus short, 504 long, 25“ in diameter. Ovary spherical or slightly oval, in 
median line or just left of it, about three eighths of body length from anterior 
end; 0.117 by 0.147 mm. in the smaller specimens and 0.147 by 0.176 mm. in the 
largest; long axis parallel to that of body. Receptaculum seminis present; 
Laurer’s canal opens just caudad of ovary. Uterus extends posteriad on left 
side, returning on right, rarely descending and ascending limbs cross about 
one third of distance from ovary to cephalic testis, forming a figure 8. Coils 
of uterus occasionally overlap diverticula through central half of distance 
from ovary to cephalic testis on one or both sides. Metraterm almost straight, 
extending caudad from genital pore one fourth to one third of distance, to 
ovary. Vitellaria arranged in lobes; separate lobes often not distinct; begin 
about one-third of distance from ovary to acetabulum, cephalad of posterior 
end of cirrus sac; extend about five sixths of distance from ovary to cephalic 
testis. Twenty to forty follicles in each lobe. Testes spherical or oval, about 
equal in size, 0.2 to 0.29 mm. in length; 0.16 to 0.24 mm. in width; separated by 
0.05 to 0.1 mm. Cirrus sac extends caudad from genital pore three fourths 
of distance to ovary; 1.12 to 1.18 mm. in length; 0.088 mm. in width. Vas 
deferens much coiled. Mature eggs average 31 by 154; those near ovary 
broader, measuring 30 by 19. 


In my material there are many immature specimens. The young 
worms are proportionately wider than the adults with the region of 
greatest width at the pharynx. The smallest mounted measured 0.65 
mm. in length and 0.16 mm. in width. In this specimen the oral sucker 
is 0.085 mm. in diameter and the acetabulum is very small, 0.03 mm. in 
diameter. The esophagus is longer and the ceca are larger propor- 
tionately than in the adult. The cirrus sac, ovary and testes appear 
merely as masses of heavily staining cells. In a specimen 1.7 mm. long 
the body is 0.2 mm. in width. The suckers have increased in size, the 
oral to 0.09 and the acetabulum to 0.063 mm. in diameter. The intestine 
has acquired the shape characteristic of the adult, the ovary has 
assumed definite form, the testes have become more prominent, and the 
cirrus sac is well defined at the anterior end although the posterior end 
is extended as a line of deeply staining cells reaching to and apparently 
connected with the ovary. No trace of uterus or vitellaria could be 
distinguished. 


NOTES ON THE TREMATODE GENUS TELORCHIS 63 


A comparison of T. corti with T. aculeatus and T. solivagus, the 
species which it most closely resembles, shows that the forms are about 
the same length ; T. corti is narrower and thicker than the others. The 
oral sucker and pharynx are smaller in T. corti and the esophagus is 
shorter. Odhner (1902) says that in T. solivagus the descending and 
ascending limbs of the uterus cross to form a figure 8; in T. corti both 
crossed and parallel conditions are present. In T. aculeatus and T. 
solivagus the cirrus sac extends from the genital pore caudad to the 
ovary, in T. corti it extends only three fourths of the distance to the 
ovary. In T. aculeatus the lobes of the vitellaria are more separate and 
distinct than in T. corti, and in T. solivagus they are not definitely 
arranged. In T. aculeatus they do not extend as far anteriad or 
posteriad as in T. corti. The eggs of T. corti are about the same size 
as those of T. solivagus and smaller than those of T. aculeatus. 

Some fifty individuals of this species, most of them immature, were 
found in the intestine of seven specimens of Malacoclemmys leseurii 
from Newton, Texas. An adult worm was obtained from the intestine 
of a single specimen of Chrysemys elegans from the same region. The 
species was also collected in June, 1910, at Havana, Ill., from the 
intestine of Malaccoclemmys geographicus. 

This species was named in honor of Dr. W. W. Cort. 


Telorchis lobosus sp. nov. (Fig. 3) 


Adults 1.67 to 2.6 mm. in length; 0.27 to 0.37 mm. in width, greatest width 
near center of body. Acetabulum circular, about two sevenths of total length 
from anterior end, in mounted specimens from 0.117 to 0.18 mm. in diameter. 
Pharynx 55 to 634 in diameter. Ovary oval, median, crosswise of the body, 
midway between anterior and posterior ends; from 40 to 70# in shorter and 
74 to 854 in longer diameter. Small receptaculum seminis present, Laurer’s 
canal passes directly dorsad, opening just above the ovary. Follicles of vitel- 
laria massed together closely; lobes distinguished only at ends of areas. On left 
side vitellaria extend anteriad about one third of distance from ovary to 
genital pore, and posteriad about seven eighths of distance from ovary to 
cephalic testis. Vitellaria of right side do not extend so far anteriad or pos- 
teriad as those on left. Normal eggs measure from 18 by 32 to 19 by 36#. 
Testes oval, lobulated; their long axis perpendicular to that of worm; from 
0.1 by 0.05 to 0.13 by 0.08 mm., very close together but not overlapping in any 
case. Caudal testis about its own length from posterior end of body. Cirrus 
sac extends from genital pore caudad to ovary; vas deferens not coiled as 
much as in T. corti. 


In size and structure T. lobosus agrees most closely with T. nema- 
toides, but a comparison with the description of Miihling shows specific 
differences. The forms are similar in the extent of vitellaria, cirrus sac, 
and size of eggs; but T. lobosus is smaller, shorter and flatter than T. 
nematoides, the suckers and pharynx are smaller, the esophagus is much 
shorter, and striking differences are noted in the ovary and testes. 

Nine worms of this species were obtained from the intestines of 
two specimens of Chelydra serpentina from Walker, Iowa. 


64 nie HOURNAL “OF PARASITOLOGY 


Telorchis medius sp. nov. (Figs. 2, 7) 


Sexually mature worms 3 to 5.28 mm. in length; 0.35 to 0.48 mm. in width; 
body closely resembles 7. corti. At anterior end cuticular spines 2.84 in 
length, in rows 34 apart. Acetabulum one fifth to one fourth of body length 
from anterior end; in mounted specimens longer than broad, measuring 0.1 
by 0.11 mm. to 0.12 by 0.146 mm. Oral sucker circular or slightly oval, with 
either diameter greater, varying from 0.115 to 0.146 mm. Short prepharynx 
shows in well-extended specimens, in sagittal sections measuring 40# in 
length. Pharynx broadly oval, 50 to 604 long and 60 to 70# broad, followed 
by long esophagus measuring 0.2 to 0.27 mm. Diverticula extend posteriad to 
point midway between caudal testis and posterior end of body, diameter 
12 to 154. Ovary spherical or broadly oval, median, three sevenths of total 
length from anterior end; diameter 0.15 to 0.2 mm. Figure 7 shows relations 
of female genital apparatus in ovarian region. Vitellaria extend anteriad to 
point midway between caudal end of cirrus sac and cephalic margin of ovary, 
and posteriad seven eighths of distance from ovary to cephalic testis; lobes 
usually distinguishable. Eggs 21 by 434; near ovary slightly more spherical, 
measuring 26 by 424. Testes spherical or oval, 0.185 to 0.25 mm. in diameter. 
Cirrus sac extends from genital pore caudad four fifths to five sixths of dis- 
tance to ovary. Vas deferens coiled more than in T. lobosus and less than in 
T. cortt. 


T. medius shows more morphological similarity to T. corti than to 
any other known species but the differences constitute sufficient ground 
for separation. TJ. medius is shorter, more uniform in width, has a 
much longer esophagus, the cirrus sac extends farther caudad, the 
acetabulum and ovary are nearer the middle of the body, and the vitel- 
laria do not extend so far anteriad, the eggs are larger, and considering 
the differences in the size of the worms the ovary and testes are also 
larger in 7. medius. 

Eleven mature and sixteen immature specimens were taken from the 
intestines of three dozen individuals of Aromochelys odoratus from 
Raleigh, N. C. 

Telorchis diminutus sp. nov. (Fig. 8) 


Mature worms 1.2 to 1.5 mm. in length; 0.2 to 0.25 mm. in width. Greatest 
width in acetabular region. Acetabulum circular or oval, two sevenths of 
total length from anterior end; 62 to 804 in diameter. Oral sucker circular or 
oval, 75 to 904 in diameter; pharynx from 30 to 404 in diameter; esophagus 
70 to 100 in length. Ovary spherical, 60 to 804 in diameter, caudal margin 
midway between anterior and posterior ends of body. Receptaculum seminis 
present, Laurer’s canal about 304 long, opens immediately caudad of ovary. 
Descending and ascending limbs of uterus usually interwoven and not in 
distinct fields. Vitellaria poorly developed, not arranged in lobes, extending 
from ovary posteriad two thirds to three fourths of distance to cephalic testis. 
Eggs measure 41 by 20H. Testes spherical or slightly oval, usually longer 
in the anteroposterior axis, 58 to 76“ by 69 to 924. Cirrus sac extends pos- 
teriad from genital pore almost to region of ovary; very much coiled. 


In general structure T. diminutus closely resembles T. parvus 
Braun, but the two species differ in many distinct items. They have 
approximately the same relative width and correspond closely in 
position of vitellaria and size of eggs; but T. diminutus is smaller, the 
suckers are larger, the esophagus is shorter, the ceca extend farther 


NOTES ON THE TREMATODE GENUS TELORCHIS 65 


caudad, the cirrus sac does not extend to the evary as in T. parvus, and 
in T. diminutus the collecting duct of the excretory system is dorsal to 
the testes instead of passing between them in the shape of a letter S. 
These parasites were obtained from the intestine of a single speci- 
men of Cinosternum pennsylvanicum from Raleigh, N.C. Some thirty 
worms were found, about half of which were sexually immature. 


Telorchis robustus Goldberger ( Fig. 6) 


The species was described by Goldberger in 1911 from a single specimen 
which was taken from the intestine of Cistudo carolina in Maryland. I have 
fifteen specimens collected in 1910 from the intestine of Chrysemys elegans. A 
study of these worms affords information which corrects and completes the 
description of Goldberger. Specimens 6 to 13 mm. in length; 0.8 to 1.3 mm. in 
width. Acetabulum 0.2 to 0.24 mm. in diameter, located about one fifth of total 
length from anterior end. Oral sucker usually slightly longer in antero- 
posterior diameter, 0.25 to 0.28 mm. in the larger specimens. Short pre- 
pharynx; pharynx 0.14 to 0.17 in diameter; esophagus short, apparent in well- 
extended specimens. Testes spherical or slightly oval, 0.4 to 0.5 mm. in 
diameter. Female reproductive organs as described by Goldberger except 
‘that Laurer’s canal opens some distance posterior to the ovary and the vitel- 
laria do not “extend in intercecal areas,” but lie in extracecal margins of body. 
Eggs (not mentioned by Goldberger) average 15 by 29x. 


Telorchis aculeatus von Linstow (Fig. 5) 


The material consists of 13 specimens collected from the intestine 
of Tropidonotus grahami in June, 1910. 

A careful detailed comparison of the worms with the description 
and figure of T. aculeatus given by Braun (1901) leaves little doubt as 
to their specific identity and regardless of the wide difference in distri- 
bution and host I shall assign the specimens to that species. 


SUMMARY 


The study of abundant material, both adult and immature forms 
from the trematode genus Telorchis, including four species new to 
science and others from new hosts and localities, has given data for 
the first general discussion of the genus yet made. The sub-genera 
Telorchis and Cercorchis proposed by Lihe intergrade and can not be 
retained. T. angustus and T. leptus if correctly described should be 
removed to an independent genus. The new and some older species 
are discussed in detail. 

LITERATURE CITED 


Barker, F. D., and Covey, G. W. 1911. A New Species of Trematode 
from the Painted Terrapin, Chrysemys marginata Agassiz. Neb. Univ. Stud., 
11 : 193-218. : 

Braun, M. 1901. Trematoden der Chelonier. Mitt. Zool. Mus. Berlin, 
2: 13-20. 

Goldberger, J. 1911. On Some New Parasitic Trematodes of the Genus 
Telorchis. Hyg. Lab. Bull., 71: 36-48. 


66 THE JOURNAL OF PARASITOLOGY 


Looss, A. 1899. Weitere Beitrage zur Kenntnis der Trematoden-Fauna 
Aegyptens. Zool. Jahrb., Syst., 12: 521-784. 

Lithe, M. 1899. Zur Kenntnis einiger Distomen. Zool. Anz., 22: 524-539. 
1900. Ueber einige Distomen aus Schlangen und Eidechsen. Centr. Bakt., 
Abt. I, 28: 555-566. 

Molin, R. 1859. Nuovi myzelminthi raccolti ed esaminati. Sitz-ber. K. 
Akad. Wiss., Wien, math-naturw. Cl., 37 : 818-854. 

Miuhling, P. 1898. Studien aus Ostpreussens Helminthenfauna. Zool. Anz.. 
21: 16-24. 

Odhner, T. 1902. Trematoden aus Reptilien nebst allgemeinen system- 
atischen Bemerkungen. K. Sven. Vetensk-Akad. Fo6rh., 59: 19-45. 

Stafford, J. 1900. Some Undescribed Trematodes. Zool. Jahrb., Syst., 
13 : 399-414. 

1905. Trematodes from Canadian Vertebrates. Zool. Anz., 28: 681-694. 

Stossich, M. 1895. I distomi dei rettili. Boll. Soc. adriat. sci. nat., 
16: 213-293. 


EXPLANATION OF PLATE 


ABBREVIATIONS 
a acetabulum p prostate gland 
cs cirrus sac ph pharynx 
e esophagus sg shell gland 
ep excretory pore sy seminal receptacle 
g genital pore sv seminal vesicle 
i intestine t testis 
1 Laurer’s canal u uterus 
m metraterm v vitellaria 
o ovary ur vitelline receptacle 
od oviduct vt vitelline duct 


os oral sucker 
All drawings are from camera lucida tracings except Figure 7, which is 
from a reconstruction. 


Fig. 1—Terminal section of the genital ducts in T. corti, showing cirrus 
sac and metraterm. 


Fig. 2——T. medius, ventral view, x 28. 
Fig. 3—T. lobosus, ventral view, x 36. 
Fig. 4.—T. corti, ventral view, x 18. 
Fig. 5.—T. aculeatus, ventral view, x 18. 
Fig. 6—T. robustus, dorsal view, x 15. 


Fig. 7—Female genital apparatus, 7. medius, from reconstruction of 
frontal sections, x 125. 


Fig. 8—T. diminutus, ventral view, x 56. 


PLATE 1 


bas 


5 
P23 SLRS 


i} 


009° 06 9.009,5 ~ 

6 6500062 9P, oo 
Fe $$ 0802862 Fn 95 %,2 09, 
Poo 09° S908) Seta Te Oe 


alo 


Figs. 1-8 


—e 


* 


fee, Nemo veces OF UTA, A PERUVIAN 
DISEASE 


Cuarces H. T. TowNsenp 
Bureau of Entomology, Washington, D. C. 


The disease commonly called uta in Peru is largely lupus vulgaris 
or tubercular lupus. It is this form apparently, or a complication 
with it, which results, when the infection becomes far advanced, in 
such disastrous cases as that of Perry Boyd, photographed on page 5, 
volume 5, of the /nca Chronicle (Cerro de Pasco, Peru; October, 
1913) and reproduced in part on Plate 36, Figure 2, of the Harvard 
School of Tropical Medicine’s report of 1913 expedition to South 
America. 

The true uta without tubercular complication is evidently not a 
more serious affection than oriental sore, to which it is very closely 
allied. So far as we yet know it appears to be confined to the Andean 
region, occurring chiefly on the western slopes in Peru, though the 
name uta is also applied in certain districts of the eastern slopes to a 
similar affection. Probably the chief endemic focus of wta is the town 
of Otao, situated in the next canyon north of the Rimac valley and 
about opposite the point where Verrugas Canyon opens into the latter. 
Ugaz demonstrated the inoculability of uta in 1886. 

Dr. Albert L. Barton, of Lima, Peru, evidently was the first to 
arrive at a correct diagnosis of wta as dermal leishmaniasis. Dr. Barton 
has shown to the writer his notes, made in 1910, recording the dis- 
covery of the specific organism in a case treated by him, then and 
there identified by him as Leishmania. These notes were not published, 
due both to lack of time from professional duties and to the belief 
that the organism was the same as that of oriental sore (Leishmania 
tropica Wright) already recorded and described. 

Jan. 3, 1913, Dr. L. Velez Lopez announced in the local press 
of Lima that he had discovered in uta lesions what he called the 
“cuerpo leishman peruvianum.’ Jan. 13, 1913, Dr. E. Escomel 
announced that this was a new form, for which he proposed the name 
Leishmania americana (Lav. and Natt.-Larr., 1912) var. uta (Cronica 
Meédica, Lima, 30: 414). 

July 7, 1913, Gastiaburt and Rebagliati announced to the Academia 
Nacional de Medicina of Lima that they had found Leishmania in uta 
lesions (Crénica Médica, 30: 324). The organisms were shown to 
the writer at the time by the authors. These findings have been still 


68 THE JOURNAL OF PARASITOLOGY 


further verified by Strong et al (Report of Harvard 1913 expedition, 
p. 178).* The nature of the affection has therefore now been 
abundantly demonstrated. 

The question of distinctness of the leishmaniases occurring on the 
eastern slopes of the Andes and in the low-lying forested country 
adjacent thereto is still open. Various names have been applied to this 
class of infection in different regions and districts. Espundia, tiacc- 
araiia, juccuya, quecpo, llaga, apaicha, huaspi, and ulcera de los bosques 
may be largely different names for the same infection. The term 
espundia is commonly applied in the montanya of Peru and Bolivia 
along the east slopes and base of the Andes, as well as farther north in 
the moist montanya. In the Pangoa montanya of Peru, the term /laga 
obtains. In the montanya of Paucartambo the affection is called 
juccuya; and in Apurimac, quecpo. In the Urubamba valley of Cuzco 
department the term tiacc-arafa obtains. In the lower tropical rain- 
forest region of eastern Peru similar infections go by the names of 
apaicha, huaspi, and ulcera de los bosques, according to locality. These 
last are described as superficial ulcers of the skin, which begin with 
itching roseate spots, the small acne-like tumors that result being 
painful. It should be noted that this description does not agree with 
that of oriental sore, the lesions of which are said not to be painful. 
Furthermore, the lesions of true uta are not painful. 

The natives of Convencion province below Cuzco say that tiacc- 
arana is caused by the bite of a “minute spider” (arafita), whence the 
name. The “aranita” is probably a larval tick, less likely a Trombidium, 
but in either case is not necessarily the carrier of the infection. In 
other parts of Peru, the natives describe the carrier as a small hairy, 
whitish fly, which is called by them uta and uta venenosa. 

Laveran and Nattan-Larrier demonstrated Leishmania in January, 
1912, in smears from lesions of a case diagnosed as espundia, sent them 
by Dr. Escomel from Arequipa, Peru (Bull. Soc. path. exot., 5:176), 
and proposed the name Leishmania tropica var. americana for the 
form. Wenyon confirmed these findings almost simultaneously in a 
case of espundia from Tambopata on the lower Rio Inambari of Peru 
(Jour. London Sch. Trop. Med., I, No. 3). Dr. Carlos Monge M. 
has also given full particulars of his own findings of Leishmania 
americana in cases of espundia and tiacc-araia in 1912 and 1913 
(Informe al Ministerio de Instruccion del Peru, 1912; Crénica Médica, 
Lima, April 30, Oct. 15, Nov. 30, 1913). 


*It is well to call attention to the statements with reference to uta on page 
6 of this report: “Its etiology hitherto had not been determined. We were able 
to show that uta is due to a species of Leishmania.” And on page 178: 
a the parasite discovered by us as the etiological factor of uta.” The 
authors have overlaoked the earlier findings—C. H. T. T. 


THE INSECT VECTOR OF UTA 69 


It is not very probable that the true uta of the western face’of the 
Andes is identical with the @spundia, tiacc-arana, juccuya, quecpo and 
llaga of the eastern slopes. It is still less likely that it is the same 
as the bouba (also wrongly spelled buba) or oral leishmaniasis of 
southern Brazil and northern Paraguay, known in the tropical forests 
of Brazil since 1759. The latter has been described by Splendore, who 
states, however, that it is undoubtedly to be identified with the espundia 
of Peru (Bull. Soc. path. exot., 5:436, 1912). Its specific organism 
has been named Leishmania brasiliensis by Vianna (Mem. Inst. 
Oswaldo Cruz., 6:41, 1914). The infection is stated to be contracted 
in the daytime in the forests of southern Brazil, and is believed to 
enter at any insect bite, or even at thorn scratches or other abrasion 
of the skin, though tabanids are indicated as the most probable and 
frequent agent of transmission. It is more likely that the Brazilian 
leishmaniasis is identical with the apaicha, huaspi and ulcera de los 
bosques of the low forest region of eastern Peru, rather than with the 
forms occurring higher up in the Andean valleys. ; 

On the night of the discovery of Phlebotomus verrucarum Town- 
send, June 25, 1913, at San Bartolome, just below the mouth of 
Verrugas Canyon in the Rimac valley of Peru, I took some thirty 
specimens of Forcipomyia utae Knab on the inside of window panes 
of the railway station. These were placed in citrated artificial serum 
as captured. Twenty-seven specimens of the Forcipomyia were ground 
up finely in 2 c.c. of the citrated serum, warmed over flame, and 
injected into the ventral region of a guinea-pig, June 27, 1913, at 
3:45 p. m., in the verruga laboratory at Chosica. Two injections of 
1 c.c. each were made at points close together. This experiment was 
numbered 20. The pig was a male, born in the laboratory at Chosica, 
May 13, 1913, of parents from Jauja, Peru. The record of the 
experiment and material secured therefrom are as follows: 


June 2/.—Injection of 27 Forcipomyia utae as above detailed. Smear made 
from bodies of 4 gnats of same species from same lot, and numbered BS 22. 

July 3.—Sore forming at point of injection. 

July 5—Sore about 1.5 cm. in diameter, subrounded, not raised, inflamed 
on edges, scabbed over. 

July 6—Scab loosened easily, lifted at one side and smear made from 
exudation beneath, scab being let back in place. Scab dark colored, sore seems 
healing slowly. Smear numbered BS 23. 

July 11—Scab came away completely, and sore is seen to be covered over 
with a thin membranous epidermis. P 

July 15—Sore healed over completely, but a small red pinhead papule has 
appeared at periphery of healed area on one side. This papule was well raised, 
1.5 mm. in diameter, and reddish or pink. Left for further developments. 

July 21—Pig dead and discarded. Temperatures from June 3 to July 17 
had been normal. 


70 THE JOURNAL OF PARASITOLOGY 


It was recognized throughout that this was not verruga, the smear 
from the sore seemed on examination to show nothing tangible, and the 
experiment was at the time deemed of no importance. The pressing 
requirements of the verruga investigation, especially the securing of 
wild flies of the Phlebotomus for injection in laboratory animals, pre- 
vented the proper study of this experiment. The two smears were not 
thoroughly examined till long afterward (November, 1913), when it 
was found that both showed a few bodies resembling Leishmamia. 
The small pinhead papule at periphery of the healed area was then 
realized to have been quite certainly uta, and it probably contained 
numerous Leishmania. It was evidently of the same nature as the 
minute red papules described by Wenyon as appearing at the periphery 
of healed areas resulting from inoculations of oriental sore in man 
(ParasitoLocy, 4: 279), and by others as following such inoculations 
in experimental animals. 


® 0 
rid ©) 6 © 
® 
Fo 9? Sp ee 
A B C D 


Aug. 15, 1913, two females of Forcipomyia townsendi Knab came 
to light in my room in the railway hotel at Matucana, Peru, where I 
was engaged in securing nocturnal bloodsuckers for study in connec- 
tion with the verruga investigation. They were placed at once in 
Gilson’s fluid for fixation, later imbedded and sectioned Nov. 6, and 9, 
1913, the four slides being numbered Sn. 20 to 23 inclusive. These 
showed Leishmania that could hardly be doubted, and caused the 
restudy of the two smears already mentioned. 

Even with these findings I have allowed the results of this experi- 
ment to rest thus far unannounced, due to doubt of the organism being 
that of uta. There seemed always the possibility that the organism 
was merely a stage of a Herpetomonas confined to the gut of the gnats. 
Furthermore, Forcipomyia townsendi, the species which was sectioned, 
occurs at Chosica, which point was supposed to be outside the uta 
zone, though this may be doubted. At the time, however, I regarded 
uta as never occurring at Chosica, which threw further doubt on the 
findings. 


THE INSECT VECTOR OF UTA 71 


Recently I have gone anew over both the material and my original 
notes of 1913. As a result believe that I have substantial proof in 
the slides (smears and sections) of the transmission of Leishmania 
uta Escomel by both species of the Forcipomyia. The data are as 
follows: 


Smear 22 (bodies of 4 Forcipomyia utae from San Bartolome).—Contains 
several forms of the Leishmania, as shown in Figure A. A minute flagellate 
form is mentioned in my original notes as found in this smear. 


Smear 23 (exudation from sore of pig 20, injected 9 days previously with 
27 Forcipomyia utae from San Bartolome of same lot as preceding ).—Forms 
of Leishmania with trophonucleus dividing, shown at Figure B. Very scarce. 


Section 20 (longitudinal sections of whole body of 1 female Forcipomyia 
townsendi from Matucana).—Shows numerous Leishmania in the abdominal 
region, apparently only in the gut; none to be seen in thoracic region or head. 
Oval to slightly pointed at one end, as shown in Figure C. Several of a large 
pointed flagellate form are mentioned in my original notes as found attached 
to wall of rectum. ; 

Section 21 (same as preceding) did not result well. 

Section 22 (transverse sections of abdomen of the other female Forcipomyia 
townsendi from Matucana)—Shows quite numerous Letshmania in various 
stages in gut, some of which are dividing, shown in Figure D. 

Section 23 (debris from sections of 22).—Shows developmental stages of 
Leishmania, 


The Leishmania is evidently voided by the gnats from the anus 
while feeding, and infection must take place when the bites are rubbed. 
Apparently there is no other possible method in which this organism 
can be transmitted by biting insects, since it exists in no form that 
can reach the salivary glands or proboscis. 

The geographical range of ufa on the west slopes of the Andes 
coincides quite well with that of the two species of Forcipomyia, which 
were found to occur from Chosica to Matucana, and may extend 
higher than the latter point. The districts in this range come mainly 
under the head of temperate sierra valleys, and include the deep humid 
quebradas of the Andes noted since ancient times as the seat of the 
disease. The seasonal prevalence of uta, given as November to April 
or the rainy months, coincides with the period of greatest prevalence 
of the Forcipomyie. 

Two cases were noted by me and clinically diagnosed as uta infec- 
tion, the first of which shows a history quite certainly traceable to the 
bites of the Forcipomyie, the second being in all probability due to 
the same cause. It was not possible to take smears from these cases 
at the time that I saw them, hence they lack microscopical findings. 

Messrs. Chadwick and Holstein, of the Peruvian Central Railway, 
lay over night in their private car on the tracks at San Bartolome about 
March 20, 1913, and have testified that many very small gnats, both 
the Phlebotomus and others, entered their car on this occasion and 


72 THE JOURNAL OF PARASITOLOGY 


bit the inmates. Mr. Gutierrez, of Lima, a minor official of the road 
and an intelligent and educated man, was one of the inmates. He 
states that he was bitten on the hands; that he saw some of the gnats, 
as at least a part of the bites was received early in the evening before 
he retired; that the gnats that bit him while he was awake were of a 
dark color, as seen by the lamplight, wings not white, smaller in size 
than the Phlebotomus, but rather stouter than same and with shorter 
legs. Following this experience, there appeared at the bites spreading 
sores. I examined these sores March 29, 1913, and can testify that 
they bore every appearance of tropical ulcer. They were eight or ten 
in number, irregularly rounded, 0.5 to 1.5 cm. in diameter, inflamed at 
edges, not raised, faintly scabbed. I prescribed citrine ointment. The 
sores lasted fifteen days or so, then healed. When I saw the case 
again on July 1, 1913, three months after, the sites of the sores showed 
as purplish scars. 

The second case was that of Dr. ———,, assistant physician at the 
Cerro de Pasco Hospital, a native of Canada, who spent some days 
and nights in the railway hotel at Matucana, about March, 1914. He 
remembered being bitten at night, did not see the insect, supposed it 
to be a mosquito, but acknowledged that he had seen no culicids. As 
a matter of fact, they were absent. A half dozen or more sores 
appeared on his wrists and forearm, not so large as in the case of 
Mr. Gutierrez, and not of such a virulent appearance. They were about 
0.5 to 0.8 em. in diameter, well raised, well scabbed, the scabs being 
dark, not spreading and not inflamed, but they persisted for many 
weeks. longer than in the preceding case, finally disappearing. 

Both of these cases I regard as mild infections of true uta, without 
tubercular or other complication. Such cases demand the same treat- 
ment as oriental sore, and are no more serious; perhaps not as serious 
as that affection. Citrine ointment, applied promptly to all bites and 
skin abrasions in the uta zones, will probably effectually prevent any 
advanced development of such infection. Dr. J. Leonidas Samanez 
states that the treatment of the developed lesions by applications of 
albuminate of mercury is superior to all others (Crénica Médica, 
18: 89-90, 1901). More recently neosalvarsan has been claimed by 
Almenara as a specific against the organism in the general circulation, 
and necessary to prevent its spread from advanced lesions to new locali- 
ties (Cronica Médica, 30: 476-7, Nov. 30, 1913). 

The incubation period of uta appears to be shorter than that of 
oriental sore. Note that only eighteen days elapsed between injection 
and appearance of the pinhead papule in above Experiment 20. The 
disease, when uncomplicated with other infection, may also run its 
course in much less time. 


THE INSECT VECTOR OF UTA 73 


That other insects than the Forcipomyie transmit Leishmania in 
Peru is very probable. Certain tabanids probably carry the organism 
of apaicha in the eastern rain-forest region. At Chachapoyas, Ama- 
zonas province, Peru, the natives accuse Ornithodoros n. sp. aff. 
turicata Duges (det. Nuttall and Warburton) of producing uta by its 
bites. This tick, like O. talaje in Mexico, inhabits the mud walls of 
the dwellings and sallies forth at night to bite the inmates after the 
manner of the bedbug ; it is probably not concerned in this transmission. 
Simulium appears never to be implicated; likewise Stomoxys seems 
excluded. Both are too generally distributed. Fleas have been shown 
to be possible carriers of Leishmania; but fleas, bedbugs and ticks 
seem excluded in the case of uta and other Peruvian leishmaniases, 
since the lesions are practically confined to the exposed parts of the 
body—hands, face and feet. Culicids are contraindicated, since all 
experiments tend to show that Leishmania degenerates in their gut. 
At all events, it appears certain that Forcipomyia utae and PF. townsendi 
are implicated as vectors of true wta on the western side of the Andes. 

In conclusion, attention should be called to one important point. 
As late as 1915, investigators of leishmaniasis have questioned whether 
the specific organism is not really a stage in the development of a 
Crithidia or Herpetomonas normally confined to the gut of insects, 
normally conveyed from insect to insect, and only accidentally trans- 
ferred to man. In the present case the fact that most species of 
Forcipomyia are normally insect-biters, attacking caterpillars and cer- 
tain other insects, would tend to confirm this view. Forcipomyia utae 
and townsendi are very abundant at times during the humid season. 
It may easily transpire that these gnats, while normally confining their 
attacks to other insects, have become accustomed, during their periods 
of greatest abundance, to transfer their attacks to man, due to a 
shortage of food-supply in the insect fauna requisite for the needs 
of their increased numbers. 


SUMMARY 


(1). The disease known as uta, occurring on the west face of the 
Andes in Peru, has been proved to be due to a Leishmania. 


(2). Two species of Forcipomyia, native to the western Andean 
region, appear to be proved capable of transmitting the Leishmania of 
uta. 


(3). It is highly probable that the various forms of leishmaniasis 
thus far known are due to as many species of herpetomonads originally 
parasitic in the gut of the insect-carriers concerned, and that, with 
regard to the occurrence in man, these herpetomonads are as yet in 
the stages of parasitism ranging from habitually abnormal or frequent 
to merely accidental or infrequent. 


FIEARTA> CYINGULA PARASITIC IN “TRE Seen -or 
CRYPTOBRANCHUS ALLEGHENIENSIS* 


FREDERIC H. KRECKER 


So far as I am aware the only worms reported to be parasitic in 
the skin of the members of the genus Cryptobranchus are an encysted 
Bothriocephalus larva found by Leuckart and a Nematode, Filaria 
cingula, mentioned by von Linstow. Both parasites infested Crypto- 
branchus maximus. 

The description of Filaria cingula given by von Linstow is taken 
from a single specimen and is rather incomplete, nevertheless his speci- 
men and a worm found by me in Cryptobranchus allegheniensis appear 
to have certain points in common. The most important difference is 
that Filaria cingula has a vulva whereas I have not yet been able to 
discover one in the worms from Cryptobranchus allegheniensis. My 
specimens are not as perfect as they might be so it is possible that 
further search will disclose the presence of a vulva. In the meantime, 
for the sake of conservatism, it may be best to consider the specimens 
as being specifically identical with Filaria cingula. It is worthy of note 
that the Cryptobranchus maximus in which von Linstow found Filaria 
cingula came from Japan whereas the host of the worms here under 
discussion is found in the Ohio river. In view of von Linstow’s 
meager description some account of these parasites may be of value. 

Six specimens of Cryptobranchus were taken from the Ohio river 
at Marietta, Ohio, in March, 1914, and kept in an aquarium. Within 
the next two months worms were found on four of the animals. They 
were threadlike, 15 to 25 cm. long, and whitish in color. The oldest 
individuals are little more than cuticular tubes filled with living young. 
These older worms protruded for varying distances from the skin of 
the host. Younger individuals did not protrude. The embedded por- 
tions of the worms were barely covered by the epidermis of the host. 
They caused a loosely sinuous, light colored elevation of the skin, the 
course and extent of which differed in each case. The worms were 
located at various places on the dorsal, lateral and ventro-lateral sur- 
faces of the trunk, on the tail, and even on the head. 

The more detailed description about to be given is based in large 
part upon the best specimen at hand which I shall hereafter refer to as 
the complete specimen. This worm is 20 cm. long. It has a practically 
uniform diameter of 0.53 mm. The anterior end is bluntly rounded 


*Contribution 44, Department Zoology and Entomology, Ohio State 
University. 


FILARIA CINGULA 75 


(Fig. 1). The mouth is terminal. There is one dorsal and one ventral 
lip. These project anteriorlyfor 0.028 mm. and are roughly triangular, 
being 0.04 mm. wide at the base and 0.02 mm. wide at the tip. In the 
two oldest specimens I noticed two blunt hooks inserted in each lip. 
They appear as two parallel, refracting lines and may be followed to 
the base of the lip. In one of the two worms they protrude a very 
short distance from the tip of the lip. In the other individual they are 
retracted. In this condition it is hard to distinguish them because their 
refractive index is about the same as that of the lips. Inability to 
detect them in the other specimens is probably due to this cause. At 
the base of the lips the body is 2 mm. wide. It increases in width very 
rapidly up to a point 1 cm. towards the posterior where it is 0.53 mm. 
wide, which is the practically uniform diameter of the remainder of 
the worm. Near its posterior extremity the body tapers down rather 
suddenly to a diameter of 0.2 mm. which it retains for 0.3 mm. and 
then ends in a rounded point 0.075 mm. wide. The anus is terminal 
(Fig. 2). 


® ¢ 


Wi Heh A rid a ee ee 


Lp di hn Bll. 


itsiae, Qe 


Fig. 1.—Optical section of the two anterior millimeters; /, lips; », phar- 
yngeal bulb; 0, ovary; e, esophagus; u, uterus; h, hooks; r, ridges. 


The cuticula is characterized by an embossment in the form of low, 
rounded, transverse ridges with bluntly rounded ends. These are con- 
fined to the dorsal and ventral surfaces, each area being 0.42 mm. wide 
and extending the length of the body. Ina surface view of the worm 
the ridges are most evident along the sides of the body where they 
appear as lateral thickenings that project out 0.01875 mm. Elsewhere 
they are hard to discern unless the light strikes them at the proper 
angle. They vary in length, some of the shorter ones being 0.059 to 
0.09 mm. long, and the longer ones 0.18 to 0.28 mm. The width varies 
slightly with the individual, those on the complete specimen having an 
average width of 0.033 mm. They are arranged end to end in rows 
with spaces varying from 0.0187 to 0.056 mm. between them. These 
intervals are usually overlapped by a ridge in an adjacent row. The 
space between rows varies from 0.037 to 0.075 mm. The embossment 
begins at the base of the lips in the form of rounded papillae-like eleva- 
tions which become successively longer and more definitely aligned 
until at a point about 1 mm. from the base of the lips the previously 
described arrangement is attained. The ridges gradually become less 


76 THE JOURNAL OF PARASITOLOGY 


distinct on the narrow region at the posterior end of the body and the 
latter half of this region is smooth (Fig. 2). There is a smooth space 
on each side of the body 0.4 mm. wide which separates the dorsal and 
the ventral embossed areas throughout their extent (Fig. 4). At both 
ends of the body all these areas become proportionately narrower. 
Scattered over the surface of the lateral fields are a number of 
extremely minute, rounded papillae. Along both the dorsal and the 
ventral edges of each field 0.093 mm. distant from the respective 
embossed areas, there are some slightly larger papillae which are more 
or less regularly arranged in two rows. The papillae of one row alter- 
nate with those of the other. The distance between those of a row 
averages approximately 0.056 mm. The rows are about 0.037 mm. 
apart. 


UR 


Fig. 2.—Optical section of posterior end; a, anus; 1, intestine; m, muscles. 
Fig. 3.— Posterior end of complete specimen; o, degenerate ovary; 
ry, indistinct cuticular ridges. 


Fig. 4—Surface view of lateral field; J, lateral field; », cuticular papillae 
in rows. 

Fig. 5.—Cross section through uterus; du, dorsal nerve; vn, ventral nerve; 
mc, muscle cell; mf, muscle fibril; 7, intestine; 0, outer layer of cuticula; 
c, inner layer; u, uterus; y, section of young; e, epidermis; ¢, tissue of lat- 
eral field. 

Fig. 6—Junction of esophagus with intestine; e, esophagus; 7, intestine; 
ud, partially distended uterus; wi, immature uterus; b, body wall. 


There are two layers in the cuticula, an inner layer about 4» thick 
and an outer layer which is probably one fifth as thick (Fig.5). In 
surface view of the lateral field two types of very fine striations are 
visible. One of them follows the circumference of the body and occurs 
at very short intervals. The other striations are, if anything, finer 
than the preceding and run diagonally. Separation of the two layers 
shows that the outer layer is homogeneous and that striations are in 


FILARIA CINGULA 77 


the inner layer. This layer stains very deeply in Delafield’s hema- 
toxylin whereas the outer layer does not take the stain but has a slightly 
amber hue. Beneath the cuticula is a single layer of extremely narrow 
columnar epidermis cells about 0.018 mm. in height. 

The alimentary tract is the nearly straight, approximately centrally 
located tube characteristic of the nematodes. In the complete specimen 
it is displaced by the greatly distended uterus and winds in long open 
spirals. There is a narrow pharyngeal tube approximately 0.375 mm. 
long in which at a point 0.26 mm. from the mouth there is a spindle- 
shaped bulbular enlargement. This is 0.15 mm. long and midway it 
has a maximum width of 0.11 mm. Anterior to the bulb the width of 
the pharynx is 0.055 mm. Posterior to the bulb the width is 0.11 mm. 
but it almost immediately widens out into the esophagus which may 
be said to begin 0.375 mm. from the mouth. The esophagus is 0.13 
mm. wide and extends posteriorly for 15 mm. to the intestine. In cross 
section it is triangular. There is a sharp constriction of the esophagus 
at its junction with the intestine (Fig. 6). The intestine has an average 
diameter of 0.187 mm. It ends in a short rectum. In the complete 
specimen the lumen of the intestine is obliterated in both the intestine 
and the adjoining portion of the esophagus, but these structures lie 
free in the coelom. Further back the intestine is represented by a 
brown, flat band which adheres to the body wall. In the younger 
individuals the intestine is free and retains its tubular shape throughout. 

The reproductive system consists of a uterus and two ovaries. As 
previously mentioned the presence of a vagina or a vulva is still in 
doubt. The uterus when fully distended with young entirely fills the 
coelom with the exception of 2.7 mm. at the anterior end and 0.56 mm. 
at the posterior end of the body. Its wall is approximately 0.0042 mm. 
thick. In the less mature individuals the uterus is only partially dis- 
tended. This is particularly well shown in a specimen in which the 
distention begins 15 mm. from the anterior end of the body. The dis- 
tended portion is only 0.2 mm. in diameter and anterior to this it 
decreases to 0.09 mm. within a distance of 0.2 mm. The young in the 
enlarged region are the size of those in the complete specimen but in 
the zone between the narrow and the distended regions there are 
extremely small young. 

The young average 0.33 mm. in length and 0.014 mm. in width. 
Their anterior end is blunt; the posterior one-fourth of the body tapers 
rapidly into a sharp hair-like point. I saw none enclosed in a capsule 
although many of them were coiled. In most parts of the uterus they 
maintain a constant wriggling, but in places they are so tightly packed 
as to leave an impression on the body wall. Individuals liberated in 
tap water retained their activity. 


78 THE JOURNAL OF PARASITOLOGY 


At each end of the uterus there is a single, slender, tubular ovary. 
The anterior ovary of the complete specimen begins 1.12 mm. from the 
anterior end of the body and runs posteriorly in a series of coils about 
the esophagus to the uterus. It then extends forward slightly beyond 
its point of origin and then again turns back and joins the anterior end 
of the uterus which is 1.59 mm. from the anterior end of the body. The 
diameter of the ovary at its tip is 0.056 mm.; for the greater part of 
its extent it is 0.09 mm.; and at its junction with the uterus 0.11 mm. 
The posterior ovary in the complete specimen has degenerated and is 
represented by an irregular mass of tissue. 

The muscles are restricted to two broad bands, one dorsal and the 
other ventral, each corresponding in extent to the dorsal and the ventral 
embossed areas of the cuticula. In the mid-dorsal and the mid-ventral 
lines each band is interrupted by a nerve cord. In a surface view of 
the animal the muscles give the effect of longitudinal corrugations 
(Fig. 3). In cross sections of the body 36 muscle cells are distinguish- 
able in each band, there being 18 on each side of the median line. 

In the material at command no excretory tubes can be distinguished, 
but I hesitate to deny their presence entirely. The lateral field, where 
such tubes are usually found, is occupied by an apparently homo- 
geneous mass of connective tissues of a rather loose texture. This 
field has a width of 0.14 mm., or practically one-fourth of the circum- 
ference of the body. 

The nervous system was also only partly distinguishable. There 
are two longitudinal nerve cords, one in the mid-dorsal and the other 
in the mid-ventral line (Fig. 5). I could not determine the nature of 
the anterior termination of the cords in any of the worms because of 
the poor histological condition of the sections in this region. 

There are some stray notes regarding the life history which may 
be of interest. As stated before, the worms are viviparous and the 
young can survive in water. They are apparently liberated by the dis- 
integration of the parent’s body. In two of the worms that portion 
which protruded from the host had been attacked by a fungus and was 
so far disintegrated that the body barely remained intact. Young 
worms which had broken out of the uterus were crowded into the 
coelom of this region. It is an interesting fact that among the two lots 
of Cryptobranchus which I have had under observation there have 
been no specimens which showed signs of infection when they were 
received. Both lots were taken from the same place at the same time of 
the year, although one lot was taken a year later than the other. In 
both cases signs of the parasites were observed about a month after 
their hosts had been caught. At this time the worms caused an.almost 
imperceptible thread-like elevation of the skin. A month later in each 
case the parasites were clearly visible and had apparently about reached 


FILARIA CINGULA 79 


the end of their growth. Both lots had been kept in filtered water 
although the supply in each case was from a different source. The 
young parasites probably entered the skin of their hosts during the 
summer from the water in which the latter lived. 

The percentage of the individuals infested must be very great since 
of the two lots which have come under my observation, there being six 
specimens in one and four in the other, all but two of the individuals 
harbored one or more of the parasites. Apparently the presence of the 
parasites is a matter of no concern to the host. There are no evidences 
of injury or of disturbance in the tissues other than the narrow tube 
formed in the epidermis of the host. -The inner lining of this tube 
glistens and is smooth except for slight indentations made by the cutic- 
ular ridges on the parasites. 

REFERENCE 


1902. Von Linstow, O. Filaria cingula n. sp. Zool. Ann., 25: 634. 
Department of Zoology and Entomology, Ohio State University, Columbus, 


THE LIFE HISTORY OF GONGYLONEMA SCUTATUM 


Brayton H. Ransom, 
Chief, Zoological Division, Bureau of Animal Industry 
AND 
Maurice C. HALL, 


Assistant Zoologist, Bureau of Animal Industry 


In April, 1911, the writers undertook some investigations in regard 
to insects as intermediate hosts of parasites. The investigations of the 
senior author were carried on at the Experiment Station of the Bureau 
of Animal Industry at Bethesda, Md., and in the laboratories of the 
Bureau at Washington, and later at Colorado Springs, Colo. Those of ' 
the junior author were carried on at Colorado Springs, Colo. Particular 
attention was paid to the dung beetles, as it seemed evident that these 
insects in working through fresh feces as is their habit, would have the 
first opportunity to ingest eggs of worms parasitic in the digestive tract 
of cattle, sheep, and other live stock. Such beetles as species of 
Aphodius, furthermore, appeared small enough to be readily ingested 
by grazing animals, and the beetles’ habit of flight from one manure 
deposit to another offered chances for such ingestion, as the beetles’ 
flight commonly terminates on grass and herbage with which they might 
readily be swallowed by cattle or sheep. 

The dissections by the senior author of Aphodius femoralis, A. 
granarius, A. fimetarius and Onthophagus hecate resulted in the finding 
of encysted larval nematodes in the body cavity. These cysts were 
about 0.5 mm. in diameter and as many as 8 were found in one 
Aphodius and 15 in one Onthophagus hecate. One larva which was 
measured was 2 mm. in length and 50u in thickness. Viewed from in 
front the head shows a narrow mouth aperture elongated dorso- 
ventrally and surrounded by a chitinous border, which is oblong quad- 
rangular in outline with rounded corners, and measures about 12p 
dorso-ventrally and about 8 from side to side. A short distance pos- 
terior of the edge of the chitinous border are 2 sub-dorsal and 2 sub- 
ventral papillae. The chitinous border of the mouth is raised above the 
surrounding surface of the head and resembles a projecting flange 
when the head is viewed from the side. The slender pharynx is 40 
long. The esophagus measures about 1.5 mm. in length, and is differ- 
entiated into a slender anterior portion about 225, long, and a more 
granular posterior portion of somewhat larger diameter. The anus is 
about 100p from the tip of the tail. The latter is blunt and is supplied 
with two or three very small short conical processes. The excretory 


LIFE HISTORY OF GONGYLONEMA SCUTATUM 81 


pore is about 200, from the anterior end of the body. About 140u from 
the anterior end of the body the esophagus is surrounded by a nerve 
ring. 

In June a number of larvae of Aphodius spp. were examined, and 
in these were discovered some young nematodes which agreed perfectly 
with the unhatched embryos of Gongylonema scutatum. The embryo 
of Gongylonema scutatum is very distinctly arinulated in the head region 
on the side opposite the mouth. The mouth is not terminal, but is a 
triangular aperture on one side of the head, with a curved hook-like 
process projecting from it. The tip of the tail is bluntly pointed. 

The finding of these newly hatched larvae in the same host beetles 
as the encysted larvae was fairly good evidence that they were of the 
same species and, in view of the failure to find them in adult beetles, 
seemed to indicate that the eggs of the worm were principally ingested 
by the beetle while it was in the larval stage. 

In July both of the writers were in Colorado and the examination of 
Aphodius coloradensis, A. vittatus, and A. granarius showed the pres- 
ence of an encysted larval nematode exactly similar to that found at 
Bethesda. These beetles were collected from sheep manure at points 17 
miles east and 75 miles northeast of Colorado Springs. 

The observations in regard to the finding of Gongylonema larvae 
in larval and adult dung beetles were briefly alluded to in the Twenty- 
eighth Annual Report of the Bureau of Animal Industry for the year 
1911, as follows: 

“Important facts have been determined bearing upon the life history 
of the gullet worm of sheep and cattle.” 

In 1912 the encysted larval forms of Gongylonema were again 
observed in the dissection of insects in connection with other investiga- 
tions in Colorado, but no further work was carried on that year. 

In 1911 an experimental feeding was made by the senior author. 
Four or five larval Gongylonema were fed to a white mouse on May 13 
and 14 more were fed to this mouse on May 17. On June 14, 32 and 28 
days, respectively, after these feedings, the mouse was killed and exam- 
ined. No worms were found. This result tended to show that the 
parasite is not transmissible to mice and that the larvae were not those 
of Spiroptera obtusa whose larvae, occurring in the meal worm, show 
very striking similarities to those which the present writers assumed 
belonged to Gongylonema. 

In 1913 an experimental feeding was made by the junior author at 
Colorado Springs. During the two months from May 10 to July 9 inclu- 
sive, a sheep was fed a total of over 250 specimens of Alphodius colo- 
radensis, A. congregatus, A. fimetarius, A. granarius, A. inquinatus, and 
A. vittatus. A dissection of a large proportion of these beetles showed 
a very small number of them to contain encysted Gongylonema larvae. 


82 THE JOURNAL OF PARASITOLOGY 


Three of the larvae obtained on dissection were also fed to this sheep. 
This sheep was killed November 18 and was found to have 7 Gongylo- 
nema scutatum in the esophagus, together with the characteristic lesions 
showing where a few others had been at one time. A companion sheep 
kept under identical conditions for a year and a half, but not fed any 
specimens of Aphodius did not develop any infection with Gongylo- 
nema. Thirty-five lambs raised under experiment conditions very much 
the same as those of the two sheep mentioned above were killed during 
the course of the three years 1911 to 1913 inclusive, and these were all 
free from Gongylonema. This furnishes an abundance of checks in 
support of the experimental finding that species of Aphodius act as in- 
termediate hosts of Gongylonema scutatum. 

During the summer of 1914 some additional work was done in the 
investigation of this life history. Cattle weasands heavily infested with 
Gongylonema scutatum were sent in from Indianapolis, Ind., by Dr. G. 
W. Butler, the egg-bearing worms cut into small fragments, mixed with 
small quantities of bread or other food and fed to specimens of 
Aphodius and to croton bugs, Ectobia germanica. The day after the 
insects were exposed to infection in this way, empty shells of Gongylo- 
nema eggs were found in the intestine, and the following day numbers 
of free embryos were found. Other eggs were found unhatched in the 
intestine and in the feces of the insects, but these were obviously eggs 
which had not yet developed to the infective stage. These findings 
demonstrated that the eggs would hatch when ingested by the adult as 
well as by the larval Aphodius. 

The young larvae found two days after exposure to infection were 
about 250u long and were apparently increasing in size. In the course 
of a week the larvae found were very much thicker. At the end of 
two weeks the larvae were about three times as thick as the original 
embryos and were apparently on the verge of an ecdysis. At this 
time they show a complete alimentary tract. The esophagus is about 
three-eighths to four-ninths of the entire body length, and is sur- 
rounded by a nerve ring a short distance in front of its middle. The 
rectum is a well-marked structure of rather large diameter, and pos- 
teriorly is closed by a plug of tissue which projects from the ventral 
surface of the body. This plug marks the location of the anus and is 
one-sixth of the body length from the posterior end. About this time 
there is an ecdysis and the cephalic annulation is lost. 

At the end of three weeks the head is more pointed but the flange- 
like margin of the lips is not yet developed. The rectum is no longer 
prominent, but the button marking the position of the anus persists. 
The larvae are now much longer. 


LIFE HISTORY OF GONGYLONEMA SCUTATUM 83 


At the end of about a month the larvae are encysted in the final 
stage. The head has the structure described in the first part of the 
paper and the anal button is lost. In favorable specimens cervical 
papillae may be seen about half way between the nerve ring and the 
anterior end of the body. 

An experimental feeding to croton bugs of eggs of Gongylonema 
from the gullet of a hog, gave substantially the same results. The 
larvae were encysted in the final stage at the end of a month. 

A rabbit was fed with three Gongylonema larvae on one occasion 
and with two on another. Two months after the first feeding and 
one month after the second, the rabbit was killed and the mouth, 
pharynx, and stomach were examined. No worms were found. A 
guinea-pig was fed with three Gongylonema larvae on one occasion and 
three more on another. Five weeks after the first feeding and three 
weeks and two days after the second feeding it died. No worms 
were found. 

August 18 a sheep was fed eleven Gongylonema larvae from a 
croton bug and a hog was fed a croton bug containing possibly fifty 
larval Gongylonema, the larvae having been developed by feeding eggs 
of Gongylonema collected from cattle. On August 25 the same sheep 
and hog were fed more croton bug material heavily infested with 
similar larvae. The hog was killed October 17, but showed no infec- 
tion. The failure to infect the hog with the nematode from sheep and 
cattle is suggestive of a specific infectivity and strengthens the idea 
that the hog nematode is a distinct species. The sheep was killed 
November 23 and the gullet found heavily infested with Gongylonema, 
the females of which were mature and full of eggs. 

While the work noted above was in progress, a very interesting 
paper appeared, dealing with the life history of another species of 
Gongylonema. Fibiger (1913) published a note in which he stated 
that he had found in rats a gastric carcinoma etiologically related to a 
species of Spiroptera. A year later, Fibiger and Ditlevsen (1914) 
published their complete study of the worm itself and the lesions 
attributed to it. The worm in question, called by them Spiroptera 
(Gongylonema) neoplastica, should be called Gongylonema neoplas- 
ticum. Gongylonema is a well established genus and there is no reason 
to question the propriety of including this species in Gongylonema, 
notwithstanding its lack of one characteristic of this genus, namely, 
the presence of cervical papillae. It is even not impossible that cervical 
papillae may be present, as these structures are frequently very difficult 
to distinguish in some species of nematodes and may be overlooked 
in numerous specimens, finally being discovered when a specimen hap- 
pens to be turned into just the right position. 


84 THE JOURNAL OF PARASITOLOGY 


Fibiger and Ditlevsen have made an excellent study of the life 
history of this worm. It was found in the first instance in rats, but it 
appears to be communicable to rodents generally as it was transmitted 
to the following: Mus decumanus, Mus rattus, Mus musculus, Lepus 
cuniculus, Cavia cobaya. The parasite occurred in the squamous- 
celled epithelium of the anterior portion of the digestive tract, including 
the mouth, tongue, esophagus and fundus of the stomach. In these 
regions the worm gave rise to a proliferation of the epithelial elements, 
originating as a circumscribed or diffuse hypertrophy associated with 
a slight inflammation, going on to the formation of papilloma, and 
terminating in distinct carcinoma with occasional metastases. 

The eggs produced by the female worm are passed in the feces 
of the infested rodent and were first found to be ingested by Peri- 
planeta americana, but were also found infective for Periplaneta 
orientalis, Ectobia germanica and Tenebrio molitor. Twenty days 
after the ingestion of the eggs by the insects, the fully developed larvae 
are found coiled in the muscles of the prothorax and limbs. It will 
be noted that this site is different from that of Gongylonema scutatum 
larvae. The location of the embryonic and larval forms after the first 
day following the ingestion of the eggs and up to the time they are 
found in the musculature of the prothorax and limbs was not 
determined. 

It is evident from the above that the life history of the two species, 
Gongylonema scutatum and Gongylonema neoplasticum is much the 
same in that the larval stage is spent in insects, at least one of which, 
Ectobia germanica, is common to both, and that the adult worm is 
found in the epithelium of the gullet in the primary host in both cases. 
The worms differ in that the larval stage of the rodent nematode is 
found in the musculature of the insect host, while the larval stage of 
the ruminant nematode is found encysted in the body cavity. They 
also differ in that the rodent nematode commonly occurs in the tongue, 
mouth and cardiac portion of the stomach as well as in the esophagus. 
Finally, the rodent nematode has the unusual power of producing 
neoplastic changes in its primary host, while there is yet no evidence 
that the ruminant nematode is more than a rather innocuous parasite. 

The life history of Gongylonema scutatum and G. neoplasticum is 
strikingly similar to that of Spiroptera obtusa, which occurs in its 
adult stage in the intestine of rats, mice and similar rodents. 

Leuckart (1867: 113-115) and Marchi (1871) found that the larval 
development of this parasite occurs in the meal worm (larva of 
Tenebrio molitor). The eggs, which resemble those of Gongylonema 
and contain similar embryos, when swallowed by meal worms hatch out 
and release the embryos. These embryos pass through the wall of the 


LIFE HISTORY OF GONGYLONEMA SCUTATUM 85 


alimentary tract, and develop in the midst of the fat surrounding it, 
becoming enclosed in connective tissue cysts. The larval development 
is complete in about six weeks after ingestion of the eggs. The fully 
developed larva measures from two-thirds of a millimeter to nearly a 
millimeter in length. The head, as described, is supplied with two tri- 
angular papillae curved on their inner surfaces, and surrounding the 
mouth except laterally. The tip of the tail is supplied with several small 
conical papillae. The excretory pore is about 100, and the base of the 
esophagus about 300» from the anterior end of the body. 

Judging from Marchi’s description and figures one of the most 
striking differences between the full-grown larvae of Spiroptera obtusa 
and Gongylonema is that the esophagus of the former is only about one- 
third the length of the body, whereas the esophagus of the latter is 
fully two-thirds the body length. 

As a postscript it may be noted that since this paper was read at a 
meeting of the Helminthological Society of Washington, Dec. 17, 1914, 
an additional intermediate host of G. scutatum has been found, namely, 
Onthophagus pennsylvanicus. Beetles of this species collected from 
sheep pastures near Vienna, Va., during the summer of 1915 were 
found to be commonly infested with the encysted larvae. 


- 


SUMMARY 


The eggs of Gongylonema scutatum present in the feces of sheep 
and cattle infested with the adult parasite, hatch out when swallowed 
by insects of various species. 

The larvae thus released from the eggs, pass into the body cavity 
and reach the final larval stage in about a month. In this stage the 
larva is coiled into a spiral and is enclosed in a capsule about half a 
millimeter in diameter. The length of the fully developed larva is about 
2 mm. and the esophagus equals about two-thirds the body length. 
The mouth, elongated dorso-ventrally, is surrounded by a flange-like 
chitinous border. 

Sheep fed upon insects containing these larvae became infested with 
Gongylonema. A hog fed upon croton bugs artificially infested by feed- 
ing with eggs of Gongylonema from cattle failed to become infested. 
A mouse, rabbit and guinea-pig fed with Gongylonema larvae from 
beetles found in sheep manure, or from croton bugs artificially 
infested by feeding Gongylonema eggs from cattle, also failed to become 
infested. Failure to produce infestation in these various animals indi- 
cates that the Gongylonema of sheep and cattle (G. scutatum) is not 
transmissible to hogs, mice, rabbits or guinea-pigs. 

Gongylonema larvae have been found in various species of dung 
beetles collected from sheep manure, namely, Aphodius femoralis, A. 
granarius, A. fimentarius, A. coloradensis, A. vittatus, Onthophagus 


86 THE JOURNAL OF PARASITOLOGY 


hecate, and O. pennsylvanicus. They have been developed in various 
species of Aphodius and in croton bugs (Ectobia germanica) by feeding 
the eggs of Gongylonema scutatum from cattle. The feeding of eggs 
of Gongylonema from the gullet of a hog (presumably G. pulchrum) to 
croton bugs also resulted in the development to encysted larvae. 

Under natural conditions the usual intermediate hosts of Gongylo- 
nema scutatum are probably dung beetles of various species. 

The life history of G. scutatum is similar to that of G. neoplasticum 
of rats, mice and other rodents, the intermediate stage of the latter 
having been found by Fibiger and Ditlevsen to develop in roaches (Peri- 
planeta americana, P. orientalis, and Ectobia germanica) and in a beetle 
(Tenebrio molitor). It is also similar to that of another rat and mouse 
parasite, Spiroptera obtusa, whose intermediate host was found by 
Leuckart and Marchi to be the larva of a beetle (Tenebrio molitor). 


REFERENCES 


Fibiger, J. 1913. Ueber eine durch Nematoden (Spiroptera sp. n.) hervor- 
gerufene papillomatése und carcinomatdse Geschwulstbildung im Magen der 
Ratte. Berl. klin. Wehnschr., 50: 289-298, figs. 1-12. 

Fibiger, J., and Ditlevsen, H. 1914. Contributions to the biology and 
morphology of Spiroptera (Gongylonema) neoplastica n. sp. 28 pp., 4 pls. 
Kgbenhavn. 

Leuckart, R. 1867. Die menschlichen Parasiten und die von ihnen herrth- 
renden Krankheiten, v. 2, 1. Lief., 256 pp., 158 figs. Leipzig & Heidelberg. 

Marchi, P. 1871. Monografia sulla storia genetica e sulla anatomia della 
Spiroptera obtusa Rud. Mem. r. Accad. sci. Torino, cl. sci. fis. e mat., 2. s., 
v. 25, 30 pp. 2 pls. 

Ransom, B. H., and Hall, M. C. 1915. The life history of Gongylonema 
scutatum. [Abstract.] J. Parasitol., 1: 154. 

Stiles C. W. 1892. On the anatomy of Myzominius scutatus (Mueller, 1869) 
Stiles, 1892. Festsch. z. R. Leuckart, pp. 126-133, pl. 17. 


NOTE ON THE STAGE OF PIROPLASMA BIGEMINUM 
ett OCeURS. IN THE. CATTLE TICK, 
MARGAROPUS ANNULATUS 


Howarp CRAWLEY 


Assistant Zoologist, Zoological Division, Bureau of Animal Industry, Wash- 
ington, D. C. 


The organism herein described, which is believed to be a stage in the 
life cycle of Piroplasma bigeminum, was found in engorged female 
cattle ticks (Margaropus annulatus) removed from cattle in September, 
1913. These ticks, which were being used as controls in a series of 
experiments at the Bureau of Animal Industry Experiment Station, 
Bethesda, Md., on the action of arsenical dips, had been immersed in 
water, dried, and maintained in the laboratory in petri dishes. Under 
such circumstances cattle ticks ordinarily behave in the normal manner ; 
that is, by far the greater number live until oviposition is fully completed. 
In the present instance, however, certain of the lots showed an unusual 
mortality, as a result of which microscopical preparations were made 
and the organism which is the subject of the present paper discovered. 

This, as seen in the figure, is a cigar-shaped body, with one end 
pointed and the other differentiated into a sort of cap. The ratio 
between the length and breadth varied considerably, but it is probable 
that this variation is due at least in part to the exigencies of fixation. 

The cap, which is placed at what is probably the anterior end, varies 
a good deal in appearance in the different specimens. In some cases it 
has the shape of a crown with a minute, pointed process arising from 
its median portion (4). In others it consists merely of a narrow shell 
fitting over the broad end of the parasite (D), while again it consists 
of a rounded body resting upon the balance of the cell like the proto- 
merite of a polycystid gregarine (B, C). As a matter of fact, these 
parasites are strikingly like minute gregarines, and may appropriately 
be termed gregarinoids. In some cases (4, D) the substance of the cap 
was much denser and more homogeneous than that of the balance of 
the cell, whereas in other cases (B, C) this distinction was not evident. 

The cytoplasm is finely alveolar, thus following the protozoan type. 
In many of the specimens there were large vacuities, but these are prob- 
ably to be accredited to the technique. An ectoplasmic layer is probably 
present, but it was not possible to demonstrate it satisfactorily. 

The nucleus occupies a median position. It is of the vesicular type 
and inclosed within a thick and very distinct membrane. Within, a large 
rounded karyosome is always present. 


88 THE JOURNAL OF PARASITOLOGY 


Fifty specimens, selected at random, were measured. The extremes 
in length were 7p and 15y; the mean length 11.24. The distribution of 
these fifty specimens, with reference to their length, was as follows: 


Length in microns No. of Specimens 
1 
6 
] 10 
11 10 
3 
4 
4 
1 


Christophers (1907) has described the life cycle of Piroplasma canis 
in the dog tick (Rhipicephalus sanguineus). In this, one of the most 
conspicuous phases is what Christophers calls the club-shaped body, 
which he describes on pages 56 to 61, and illustrates by a number of 
figures on his plate 2. The precise resemblance between these, and the 


forms herein described from Margaropus annulatus, leave no doubt as 
to their being corresponding stages in the life histories of Piroplasma 
canis and Piroplasma bigeminum. 

According to Chistophers, the club-shaped bodies are derived 
directly from the parasites ingested by the tick in blood from the dog. 
The difference in both size and morphology between the intracellular 
form of Piroplasma canis and the club-shaped bodies is considerable, 
but Christophers figures several intermediate stages, and there seems to 
be no reason to question his conclusions. The club-shaped bodies were 
found in the gut of the dog tick, and, more abundantly, in the ovaries, 
oviducts, and eggs. Christophers describes two forms, as follows: 

“(a) Rather rigid thorn-like bodies resembling on a very small 
scale certain gregarine trophozoites. This resemblance is largely due to 
the presence of a peculiar disc-like structure armed with cusps carried 
at the anterior extremity. They are sometimes motionless, but usually 
exhibit rather slow movements, especially a side to side flap-like action 
of the tail portion. The protoplasm is transparent, slightly refractile, 


PIROPLASMA BIGEMINUM 89 


and free from large granulations, and in it a clear area (the nucleus) 
can often be made out. ~ 

“The disc has the appearance of a boring organ. It carries four 
or five cusps, one of which is situated centrally, whilst the others are 
arranged around the periphery. Immediately behind this structure the 
parasite is often constricted so that a kind of neck is formed. 

“(b) Leech-like forms more club-shaped than those just described 
and executing very active movements. They possess a swollen end 
which is often attached to the slide or cover glass and a thin end which 
is kept in constant motion like the head of a leech. They also undergo 
modified ameboid movements, the shapes depicted in plate 2, figure 2, 

® being characteristic. 

“These bodies are most numerous as a rule about the oviducts, and 
a few can be found here when not to be detected elsewhere. They also 
occur in the ovary and may be seen lying with ova not yet free in the 
lumen, and in more mature ova. By careful search they can also be 
found in fresh preparations of the gut. 

“In stained preparations a greater variety of forms is apparent, but 
the two forms mentioned, those with and those without a disc, are dis- 
tinguishable. Since intermediate stages are not difficult to find it is 
probable that both are identical in nature, the disc bearing forms being 
the more mature stage.” 

Finally, according to Christophers, these club-shaped bodies enlarge, 
become irregularly rounded, and transform themselves into what he 
designates as the zygote. This term, however, is singularly unfortunate, 
since no sexual process is described. The “zygotes” by a series of steps 
which Christophers describes in detail, break up into sporozoites, these 
later stages of the life cycle taking place in the salivary glands of the 
tick. 

Koch (1906), studying P. bigeminum, figured and described what 
are clearly the same bodies, although he says nothing about the curious 
cap or crown present at one end of the parasite. Koch found these ele- 
ments both in engorged female ticks three days after removal from the 
cow, and in the eggs. According to his account, they are the later 
rather than the earlier stages in the evolution of Piroplasma in the cattle 
tick, but he did not follow them further than the stage here under con- 
sideration. 

Kleine (1906) and Nuttall and Graham-Smith (1908) cultured Piro- 
plasma canis, and obtained developmental forms much like the earlier 
stages of P. bigeminum in the tick, as described by Koch. The cigar- 
shaped body, however, has so far only been seen in either ticks or tick 


eggs. 


90 | THE JOURNAL OF PARASITOLOGY 


These various observations are not wholly accordant, nor do they 
seem to cover the entire process. Minchin (1912: 384), however, has 
endeavored to put them together and to formulate an outline of the life 
history of Piroplasma in the tick, and to this the reader is referred. 

My own observations on Piroplasma bigeminum in Margaropus 
annulatus concern only the club-shaped bodies or gregarinoids. As 
already stated, these differ somewhat amongst themselves with regard to 
the morphology of the cap at the anterior end, but, as Christophers sug- 
gests, such differences are probably merely a matter of the stage of 
development. In my material from Margaropus annulatus, the condition 
of the cap shown in D, was quite infrequent, from which it may be sur- 
mised that these gregarinoids were in an earlier rather than a later stage 
of their evolution. Furthermore, as already stated, the gregarinoid was 
the only stage that I was able to find, or, at least, to recognize. 

In addition to their occurrence in smears made from the ticks them- 
selves, the gregarinoids were also found in preparations made from the 
crushed eggs. In the eggs, however, they were very scarce and could 
be picked up only with the greatest difficulty. In crushed seed ticks, 
hatched from eggs laid by ticks of the infected lots, I have not been able 
to find either the gregarinoids or any element which can be identified 
as belonging to Piroplasma bigeminum. 

It has been noted that the ticks in which the parasites were found 
had shown an unusually high mortality, it being on this account that 
they were examined. This suggests that the Piroplasma is pathogenic 
for the tick as well as for the cow. 

In consequence of the discovery of the parasite of Texas fever in 
the tick, a series of experiments was carried out and, although they were 
wholly negative, the failure is of itself of some significance, and a brief 
résumé may not be out of place. 

The ticks which showed the high mortality were collected early in 
September from a certain cow (No. 1040 of the Bureau of Animal 
Industry series). The microscopical preparations were made on 
September 22 from ticks collected on September 4, or 18 days after 
reaching maturity. The weather being then warm all of the ticks which 
had survived were either in the midst of oviposition or had nearly or 
quite completed it. It was obviously beside the point to search for 
the parasite in the bodies of ticks which had laid all their eggs since 
such ticks merely shrivel and die. Moreover, the material was not 
abundant. 

In consequence, the eggs of the batches of ticks (collected Septem- 
ber 2, 3, 4 and 5) which showed the high mortality, and which were 
moreover known to be infected, were kept in appropriate containers 
and permitted to hatch. On Oct. 18, 1913, microscopical preparations 


PIROPLASMA BIGEMINUM 91 


were made of a number of the seed ticks, and the balance of them were 
placed on cow 1040. Thése ticks began to come to maturity on 
November 24, and collections were made daily from November 24 to 
November 28. Microscopical preparations were made daily of these 
ticks from one to seventeen days after their removal from the cow, but 
the parasite could not be found. In consequence, the experiment was 
abandoned. 

These negative results are in line with the results of observations 
made in 1908, during the course of which a number of infectious ticks 
were examined without detecting anything which could be identified as 
belonging to the life cycle of Piroplasma bigeminum. The inference 
seems to be that of a given number of so-called infectious cattle ticks, 
only a certain proportion actually harbor the parasite. It is easy to 
understand how this might take place. A female cattle tick lays several 
thousand eggs and in order that her entire brood should be infected it 
would be necessary that each egg receive at least one parasite. Such 
a condition could fail of. realization in two ways. In the first place, 
the engorged female might not harbor as many parasites as eggs, and 
in the second, the distribution of the parasites would need to be 
remarkably accurate to insure that each and every egg became infected. 

It is also evident, from the experiments herein outlined, that 
whereas a given lot of ticks may be heavily infected, the offspring of 
these may be negative at least so far as a microscopical examination is 
concerned. 

It may finally be noted that a spirochaete has several times been 
detected in cattle ticks and that it was present in the lot which showed 
the heavy infection with Piroplasma. Spirochaeta theileri, first dis- 
covered by Theiler (1904) in cattle in Africa, is stated to be the causal 
agent of a mild disease. It is known to be transmissible by Margaropus 
decoloratus, and further that adult infected ticks of this species can 
transmit the spirochaete to their offspring. Margaropus decoloratus 
and Margaropus annulatus, the former in Africa and the latter in the 
United States, play similar rdles in the transmission of cattle diseases. 
Hence it may be suggested that the spirochaete occurring in Mar- 
garopus annulatus is Spirochaeta theileri. 


SUMMARY 


A parasitic protozoan was found in smears made from female cat- 
tle ticks (Margaropus annulatus), and from crushed eggs which they 
had deposited. The parasite has the form of a minute polycystid gre- 
garine, and is believed to represent the stage of Piroplasma bigeminum 
occurring in the tick. It is essentially like the form figured and 
described by Koch as present in engorged female ticks and their eggs, 
and also like the form of Piroplasma canis found by Christophers in 


92 THE JOURNAL OF PARASITOLOGY 


Rhipicephalus sanguineus. In the present case, it is of interest to note 
that the female ticks in which the parasites were found showed an 
unusual mortality, suggesting that the parasite is pathogenic for the 
tick as well as for the cow. In addition to the gregarinoid parasite a 
spirochaete was found in the ticks. This parasite not heretofore 
reported from the United States is perhaps the same as the form 
known as Spirochaeta theileri. 


REFERENCES 


Christophers, S. R. 1907. Piroplasma canis and Its Life Cycle in the Tick. 
Scient. Mem. Off. Med. and San. Dept. Govt. India, n. s. 29, 83 pp., charts 1-7, 
pls. 1-3. 

Kleine, F. K. 1906. Kultivierungsversuch der Hundepiroplasmen. Ztschr. 
f. Hyg. u. Infektionskrankh., 54: 10-16; 2 pl. 

Koch, R. 1906. Beitrage zur Entwicklungsgeschichte der Piroplasmen. 
Ztschr. f. Hyg. u. Infektionskrankh., 54: 1-9; 3 pl. 

Minchin, E. A. 1912. An Introduction to the Study of the Protozoa, with 
Special Reference to the Parasitic Forms. 517 pp., 194 figs. London. 

Nuttall, G. H. F., and Graham-Smith, G. S. 1908. The Development of 
Piroplasma canis in Culture. Parasitology, 1: 243-260; 1 pl. 

Theiler, A. 1904. Spirillosis of Cattle. J. Comp. Path. and Therap., 
17 : 47-55. 


SOCIETY PROCEEDINGS 


THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON 


The twenty-seventh regular meeting of the Society was held at the resi- 
dence of Doctor Hall, Oct. 22, 1915, Doctor Hall acting as host and Dr. N. A. 
Cobb as chairman. 

The following resolution was approved by the Society: 


Wuereas, Unnecessary changes in the names of host animals add to exist- 
ing difficulties and confusion in the study of the parasites of these hosts, 
therefore be it 

Resolved, That the Helminthological Society of Washington approves and 
endorses the official lists of such generic names as authorized by the Zoological 
Congress at Gratz, and urges that helminthologists use the approved names 
in their publications. 

Dr. B. H. Ransom presented the following note on a third American case 
of Dipylidium caninum in man: 


Under date of Sept. 15, 1915, Dr. L. T. Cassidy of Norwich, Conn., for- 
warded to the Zoological Division of the U. S. Bureau of Animal Industry for 
determination some segments of a tapeworm passed by a Polish child aged 
1% years. On examination the tapeworm proved to be Dipylidium caninum. 
This apparently is the third case in man to be reported from the United States. 
Stiles (1903c) reported a case in a 16-months old child at Detroit, Mich., and 
Riley (1910)* a case in an ll-year old boy at Ithaca, N. Y. 

This tapeworm is a common parasite of dogs and cats, but comparatively 
rare in man, altogether less than 100 human cases having been reported. Most 
of the cases of infestation in man have been of children less than 3 years 
old. Infection undoubtedly occurs as a result of ingesting infested lice 
(Trichodectes canis) or fleas (Ctenocephalus canis). The chances of swallow- 
ing these insects are of course greater in the case of children than of adults, 
which probably explains the greater frequency of infestation in the former. 
As an indication of the frequency of infestation in dog fleas it may be of 
interest to note that in July, 1912, I examined 21 fleas from a dog which was 
heavily infested with Dipylidium caninum; two of the fleas were found to 
harbor cysticercoids, one in one case, and seven in the other. 

Doctor Ransom also presented the following list of parasites from the 
Island of Guam: 

During the last year the Zoological Division of the U. S. Bureau of Animal 
Industry has received from Dr. L. B. Barber of the Guam Agricultural Experi- 
ment Station, a number of specimens of parasites for identification. The fol- 
lowing species were represented among these specimens: 

Trematoda—Fischoederius cobboldii (?), cow; tentative determination. 
Fasciola hepatica, cow. Fasciola sp., carabao; resembles F. gigantea in form, 
but is smaller. 

Cestoda.—Davainea echinobothrida, chicken. 

Nematoda.—Oxyuris equi, horse. Stephanurus dentatus, pig. Metastrongylus 
apri, pig. Trichuris sp., pig; apparently not T. crenatus. Tetrameres fissispinus, 
chicken. Cheilospirura nasuta, chicken. Ascaridia perspicillum, chicken. 
Heterakis vesicularis, chicken. O-xyspirura mansoni, chicken. 


* Science, 31: 349. 


94 THE JOURNAL OF PARASITOLOGY 


Arthropoda—Menopon trigonocephalum, chicken. Goniocotes gigas, chicken. 
Dermanyssus gallinae, chicken. Haematopinus tuberculatus, carabao. Margaropus 
caudatus (?), cow; tentative determination. 

Protozoa.—Theilaria parva (?), cow; tentative determination. 

Dr. C. W. Stiles presented a note by Stiles and Graves on the lung capacity 
of children in the city of X. It was found on spirometer examination of 
1,618 white school children, that from 6 to 13 years old (primary and grammar 
school age) the boys average from 100 to 200 c.c. greater lung capacity than 
the girls. From 14 to 17 years (high school age—an athletic period) the boys 
have progressively from about 300 to about 1,100 cc. greater lung capacity 
than the girls, a striking increase. 

From 6 to 13 years old inclusive, the yearly increase in the lung capacity 
of the girls is very similar to that of the boys, but at 14 there develops a 
distinct decrease of this increase, and from 14 to 17 years inclusive, the annual 
increase averages distinctly less than for the years 6 to 13. 

The decrease of the increase at 14 years in the girls follows immediately 
on the average age of beginning menstruation (13.2 years) and corresponds 
with the decrease of the increase in height (sitting and standing) and weight. 

There is a slight irregularity of the increase curve at 11 in both boys 
and girls, corresponding to the irregularity found for the same year in the 
curves for height (sitting and standing) and weight in the boys, and for sitting 
height in the girls. 

In the case of both the boys and the girls, children from homes provided 
with better sanitation (sewer) have a tendency (total, 15 to 9; boys 8 to 4, 
girls 7 to 5; estimated in year groups) to greater lung capacity than the chil- 
dren from homes with poorer sanitation (privy; total, 9 to 15; boys 4 to 8, 
girls 5 to 7). 

In cases of intestinal infection it was not evident that hookworm, Ascaris 
Lamblia, or Endameba coli has any noticeable effect on the spirometer tests. 
While pupils with whipworm infections showed a preponderance of tests lower 
than the average, the number of cases is so small that conclusions are of 
doubtful value. 

Doctor Stiles stated that findings formerly reported to the effect that chew- 
ing tobacco decreases and smoking increases with improved sanitation has 
been found to be true for city children as well as country children. 

He presented another note setting forth the fact that in a study of the 
weight and the standing and sitting heights of a number of children, those 
with light hookworm infestations were found to be below the average in 
91 markings and above in 68. 

Doctor Stiles presented the following findings in an extensive study of the 
blood of 295 boys, from 6 to 17%4 years old, and of 279 girls in the city of X: 
The red blood cells are below the accepted normal for adults, being 4,617,000 
for the boys and 4,678,000 for the girls. In 234 boys from homes with good 
sanitation (sewer), the red count was 4,633,000; in 51 boys from homes with 
poor sanitation (privy), the red count was 4,591,000. In 200 girls from homes 
with good sanitation (sewer), the red count was 4,752,000; in 74 girls from 
homes with poor sanitation (privy), the red count was 4,498,000. The lowest 
red count for a boy was 2,912,000; the lowest for a girl was 3,536,000. The 
hemoglobin averaged 86 per cent. for the boys and 87.4 per cent. for the girls. 
In 234 boys from homes with sewer the hemoglobin was 86.7 per cent.; in 
51-boys from homes with privy the hemoglobin was 84.4 per cent. In 200 girls 
from homes with sewer the hemoglobin was 87.7 per cent.; in 74 girls from 
homes with privy the hemoglobin was 87.5 per cent. The highest hemoglobin 
index among the boys was 115 per cent. and the lowest 52 per cent.; the 
highest hemoglobin index among the girls was 128 per cent. and the lowest 
40 per cent. 


HELMINTHOLOGICAL SOCIETY OF WASHINGTON 95 


Although the standard number of white cells is regarded as 5,000 to 7,000, 
the average for the 295 boys#was 8,012, and that for the 274 girls was 7,734. 
In a general way the increase in white cells is indicative of infection, poor 
condition, and the like. The 234 boys from homes with sewer showed a white 
count of 7,680; the 51 boys from homes with privy showed a white count of 
8,687. The 200 girls from homes with sewer showed a white count of 7,731; 
the 74 girls from homes with privy showed a white count of 7,771. <A differ- 
ential white count, the most extensive ever made on children, was also part 
of the study. The average count for the 295 boys was as follows: Poly- 
morphonuclear neutrophils, 52.06; eosinophils, 6.32; large mononuclears, 6.41; 
small mononuclears, 32.53; transitionals, 2.18; basophils, 0.47. The average for 
the 279 girls was as follows: Polymorphonuclear neutrophils, 53.2; eosinophils, 
5.43; large mononuclears, 5.92; small mononuclears, 32.89; transitionals, 2.11; 
basophils, 0.51. The extreme high counts for the different types of cells in 
individual children were as follows: Polymorphonuclear neutrophils among 
the boys, 79; among the girls, 78.8; eosinophils among the boys, 27.2; among 
the girls, 26; large mononuclears among the boys, 32.5; among the girls, 36.5; 
small mononuclears among the boys, 65.8; among the girls, 60; transitionals 
among the boys, 10.4; among the girls, 7.6; basophils among the boys, 2.4; 
among the girls, 7.5. The extreme low counts for individual children were 
as follows: eosinophils, none; large mononuclears, none; transitionals, none; 
basophils, none; neutrophils, 18; small mononuclears, 2. 

Details of this blood study will be published in Public Health Reports. 

Doctor Hall presented notes on the following subjects: The city of X in 
the county of Z; A case of spurious parasitism. 

Doctor Cobb exhibited a Spencer portable microscope. 

Doctor Cobb presented a note bearing on parthenogenesis. Maupas in 1900 
published a study of such nematode species as usually present only females, 
and in which the same gonad produces sperm cells and later eggs. Cobb calls 
this kind of hermaphroditism, syngonism, stating that, including the cases 
observed by Schneider, Biitschli, Claus, Maupas, himself, and others, the num- 
ber of syngonic nematode species has now risen to several scores, and that 
there are excellent reasons for believing that syngonism is very common among 
free-living nematodes—is in fact the prevailing condition in numerous large 
genera. He suggests that supposedly parthenogenetic forms in general should 
be carefully reexamined with a view to determining whether some of them 
are not actually syngonic. He exhibited a figure of one of a series of syngonic 
free-living nematodes, showing consecutive processes of sperm and egg pro- 
duction, and said that he had series of species in which the spermatozoa to 
be found are smaller and smaller until they reach the optical limits of present 
instruments and methods, so that he finds himself in the position of being 
unable to assert the non-existence of spermatozoa in certain nematodes simply 
because he does not succeed in finding them. The researches naturally sug- 
gest the possibility that small spermatozoa may in the past in some cases have 
been overlooked. 

Doctor Cobb also presented a drawing showing the mechanism of the spin- 
neret of a free-living nematode. This mechanism, not heretofore understood, 
consists of a vesicle with an outlet controlled by a “needle-valve”—not, how- 
ever, a free part as in machine valves. A somewhat needle-like plug seems to 
be forced into the outlet aperture by the pressure of the caudal secretions 
passing into the vesicle. To open the valve and permit a flow of secretions, 
the “needle” is withdrawn by retractor muscles passing from the proximal end 
of the needle obliquely forward to the dorsal wall of the tail. ‘ 

Maurice C. Hatt, Secretary. 


NOTES 


The Annual Report for 1914 of the United Fruit Company Medical Depart- 
ment furnishes interesting data for the parasitologist. The seven divisions 
of the medical service cover fairly well the territory bordering on the 
Caribbean Sea. Conditions represented in the report concern the properties 
and employees of the company and are probably distinctly better than those 
in adjacent territory not so controlled. The data covering diseases caused by 
animal parasites have been collated from the complete tables of the report 
and are presented below in tabular form. Some records are more elaborate 
or more carefully worked out than others; and the absence of any entry 
under a given heading does not demonstrate the non-occurrence of the para- 
site or the disease at that place. None the less the report gives an interesting 
and valuable survey of the prevalence of animal parasites in man within this 
area of which relatively little is known in this respect. The figures are obtained 
from a grand total of 127,809 patients of whom 15,406 were treated in hos- 


s Costa Rica |Guatemala| Colombia | Honduras! § a oy go 
8 Division | Division | Division | Division |2_|3 |Sa a 
a 2g!) ZO See hese 
es ee A~|\A~laZla= 
| Ba Slas| Sl] = 
OB) es 
qa2 ge2 AaA| A 
| n n n mn a5 | a= 
2 2 2 2 OS Oilers 
j2a| 2 Bo a | EO a ee | oe ae ee 
is2/2 | #|/2/2)8|] 8/8|8 |£8|28)32 |e 
lab) 2 2 a 2 a 2 a ® an|an|s§e |as 
an na a nD a DB Q nD a agsl\|ng|/sS/|so 
oA] 6 a a Oo} 2 | oO |] 2 | Cujouls4 ign 
| q A ae a 0) A 3) fa) qt ft @ ta 
—— | dal S| Re een — —. — — 
Waar eae 1s ele exe | 857 | 1,049 2,200 | 678 | 5,244 | 558 | 1,558 | 141 | 6,902 | 799 | 104 | 119 | 217 
Amebieldysenterys-..-\" Si) ce..s)| 11] Bi | eeeccull) 17Bh lesen pao biel lene amber taney oi" 
Myasis, nasal......... hes 2 ers ; 2_| 8| 4 
Ancylostoma.......... (too) 288] 9/406 |...) “ital 89) Bey) Ge) oe) oe 
| | | / 
Ascarisea octet 3 15 | 9 | 119 2 1 60| 8 43'\| Feat eS eee 
Strongyloides......... 1} ih Soca dh eres Til" Scracte ROA Reena nee lee eel lt oer esmeed Bcc 
Tapeworm............ | + 1 1), 2) 20 es be eaeesaieaseal see pe | es orate Magee 
Other intest. parasites, 1 28 > ee ieee eoeon MerrnG Aaron lkamree cto. | bade! Acc 2 
| | | 
Schistosoma.......... Jace | asses | sen fis ena Ihteaign| teeamey| nab esa kee sn| tea en 
Reablens. car eaedce ns 2 ao) 67) 921 a 60)| 2.) _68)| js) abel ae 
| 
ONIE Peres oe ccciccccien se ccicar| MARA ae AOA cance acts joctcas. || cec (1 As nora moe | Pcsoce |) 
| 
Myasis, skin........... eer il ares & se errr ae eoerot lee: ilttasoelPeaMallecarh |} oe | c= 
Ground itch........... 1 £4|) Spal! aS eee 8} den] S87 | ce eh aoe 
| } | 
Elephantiasis......... ese 2 | UN Sbsaipaeuce {Mec DL) aiey.|]!\seeete ||| ncsiet| eres] perce eee 
DHODIGACR. ws. seco see 4 20 | 359]... DOO” |iielaet Soliisee MSL | siete. | petsiohe. eee 7 
Tropical ulcer......... 30 19 | 54 at 9 476 1 | * a 
Venomous bites and | 
SLIME Sean ismsrcicce eer 1 32 1 42 4 1007) cas 12 24 12 
Snake bites............ ube 5 13 2 1| 2 4 | see . : 
| | | 


* Excluding cases classed as clinical malaria at hospitals. 


NOTES 97 


pitals, 91,324 in dispensaries and 21,079 on steamships. The report is a valu- 
able document and the company is to be congratulated on the organization and 
efficiency of its medical service. Similar data are not available in the large 
majority of our own highly educated and civilized states. 


CONSOLIDATED LABORATORY REPORTS 


| Pan- | Costa | Guate- Colom- 


ama Rica mala | bia Hon- Cuba Cuba 
| Divi- | Divi- | Divi- | Divi- | duras| (a) (b) 
| sion sion sion | sion 


Blood examinations.............++-+-e++0 | 2,374 7,643 2,240 780 660 892 
Estivo-autummnal............2++.0ssse00+ sae | anit 338 | 192 | 270 | 205 | 46 
MWe 5 ve haope Stake eae | 441 467 321 | 198 22 | 335 
SPARED Moye cans chacxesndapeeiepanlehtase senne 173 6 | 106 
Mie 2.555 dase: sbe, tages eacekaied a, (pane. 6 9 2 

Stool: examins Hows, 4-0 > vss uceseevabos 1971 | 6,212 | 2,996 | 428 774 | 6 
Aneylostomal............00-s0s0+ apes 124 | 1,172 879 | .... | (80%) | «.... |(70%) 
Ancylostoma and Trichocephalus....... if, wa Poe ao | 72 | 3 
Ancylostoma and Ascaris............... Ee ea ECO EOE 25 | 
Aneylostoma, Ascaris, and Tricho- 

CS a a i ES Se ARS Le 53 dae AS eee 31 
Ancylostoma and Strongyloides........ rs Say UR Pte awe / ES paces 1 
Ancylostoma duodenale................. 13200 wees saat 76 95 7 
Ascaris lumbricoides.............--...+-- 81 | 403 525 | 39 17 
Trichocephalus dispar..............+..+. 226 | 1,048 38 | 2 73 8 
Ascaris and Trichocephalus............. cat hence Ler 36 oe ee 
Asearis and Oxyuri8..........-.-c0.+0 Sh) sasee— i asane seeee | 9 . 

Strongyloides intestinalis......... seats 67 | 123 76 | 91 21 | 7 + 
Oxyuris vermicularis.......... eas sania eos 2 | 41 woitas | aa wees | wee 

RMN MOBEN Rone as <4 nsnnencescccsnsnees Jali aawaut | 1 cones | 1 Sos | bee 
Miarirmrnrinds cc o-oo. ics ncck bh caca soes'eee ae | re 3$* | 1 AP eee 1 
2 Teale 5 oe ee ee oe 34 / 156 Ap : 
a LE Se eee ee 5 | 872 $1 122t 3 - 
(ONS | Ba ee eee any eee : AD) wees 7 ee * 


Also Ancylostoma, Ascaris, and Strongyloides, 1; Ancylostoma, Trichocephalus, and 
Strongyloides, 20; Ancylostoma, Ascaris, Strongyloides, and Trichocephalus, 4; Strongyloides 
so i 4; Strongyloides, Ascaris, and Trichocephalus, 3; Strongyloides and Trichoceph- 
alus, 19. 

* One each Taenia saginata, Hymenolepis nana, Dibothriocephalus latus. 

+ With Ascaris, 1; with Ancylostoma, 121 

t Cercomonas was recorded in 9 out of 2.353 urinalyses 

§ Amebae were recorded in 1 out of 6,148 urinalyses. 

{ Apparently no differential diagnosis between the two human hookworms. 


98 THE JOURNAL OF PARASITOLOGY 


RATE OF GROWTH OF THE BEEF TAPEWORM 
IN HUMAN BEINGS 


Waite lecturer in zoology at Potchefstroom Agricultural School, South 
Africa, the author had an opportunity of determining the rate at which a 
tapeworm will grow in human beings and, as there seems to be very few 
references in literature, it was thought a short comment might be of interest. 

One of the students at the school was treated for tapeworm and passed 
most of the worm, which was 9 feet 6 inches in length. The head, however, 
was not passed, and from the width of the smallest segments passed, there 
could not have been more than 6 inches of the worm left in his body. The 
date was carefully noted, but the student was not treated until such time as 
he was again suffering from the tapeworm. When treated the second time, the 
entire worm was passed. The worm then measured 19 feet 9 inches in length, 
showing a growth of about 19 feet 3 inches in seventy-two days. On obtaining 
the head, it was determined to be the beef tapeworm, Taenia saginata. 


Witt1AM Moore, University of Minnesota. 


PSOROPTIC OTACARIASIS—PSOROPTES CUNICULI 


Full grown gray and white rabbit, female. Attention was drawn to the 
animal because of drooping ears, being the only animal among forty so 
affected. The animal was caught and examined, when it was found to be 
affected by some parasitic disease. The ears were enormously thickened and 
completely filled with dry, dirty gray crusts. At the base of the ears were 
areas free of hair and covered with thin small scabs, which on removal gave 
a raw, bleeding surface. The scabs in the ears were difficult to remove, and 
when finally removed in toto caused a good deal of pain and small punctate 
hemorrhages. The scab consisted of dry, horny, crescentic, speckled white and 
gray layers, each layer containing numerous eggs and minute parasites. By 
microscopic examination, the parasite proved to be a mite and which later, on 
closer examination, seemed to be Psoroptes cuniculi (Psoroptic otacariasis). 

The animal was thoroughly cleansed, received daily washes of hot bichlorid 
of mercury 1:200, dried, and a thick layer of carbolic vaselin rubbed over 
the ears. Complete recovery with no infection of the other animals. 


Boston, Mass. JEROME A. Honeryj. 


The Journal of Parasitology 


Volume 2 MARCH, 1916 Number 3 


LA FAMILLE DES THELAZIIDAE 
A. RAILLIET, ALFORT, Paris 


Dans l’importante superfamille des Spiruroidea, on peut dés a 
présent établir un certain nombre de coupes correspondant a des 
-‘familles. 

C’est ainsi qu’ont été constituées déja les familles des Spiruridae 
et des Acuariidae. De meme le genre Tetrameres Creplin peut étre 
considéré comme le type d’une famille des Tetrameridae; le genre 
Hedruris Nitzsch, celui d’une famille des Hedruridae; les genres 
Ancyracanthus Dies. et Ancyracanthopsis Dies. méritent d'etre groupés 
en une famille des Ancyracanthidae, etc. 

‘Il me parait utile enfin de constituer une famille des Thelaziidae 
pour les genres du type Thelazia Bosc. A cOoté de ce type viennent en 
effet se ranger naturellement les genres Ceratospira Schneider et 
Cystidicola Fischer, ainsi que deux autres genres nouveaux, Schistoro- 
phus et Serticeps. Et l'on peut méme en rapprocher provisoirement le 
genre Oxyspirura Drasche, ainsi que deux autres groupes a créer: 
Galeiceps et Rhabdochona. 


Famille des THELAZIIDAE.—Spiruroidea. Téte nue ou pourvue, 
soit d’expansions cuticulaires, soit d’un revétement en forme de casque. 
Bouche tantot sans lévres, tantot a six petites lévres, parfois a deux 
seulement, suivie en général d’un vestibule allongé ou d’une courte 
capsule buccale. C£sophage composé, dans la régle, de deux parties 
distinctes. 

Males a queue généralement obtuse, avec ou sans ailes latérales 
(bourse), portant de chaque coté wne rangée linéaire de nombreuses 
papilles préanales parfois couplées; papilles postanales peu nom- 
breuses ; 2 spicules presque toujours trés inégaux. 

Femelles a queue généralement mousse; deux utérus; vulve a sit- 
uation trés variable. Ovipares ou vivipares. 

Habitat.—Région orbitaire des Mammifeéres et des Oiseaux; tube 
digestif ou vessie aérienne des Oiseaux ou des Poissons. 

Genre type: Thelazia Bosc. 


100 THE JOURNAL OF PARASITOLOGY 


Le tableaus ci-aprés peut servir de clé pour la détermination des 
genres : 


MSOC Gs ee tN rae We his aici ea «csc osad » detam eae oe meee Galeiceps 
RC UIN nA SEA lg rato cesta a s.0 > « ss 0c ue Bi 1 ao mia ete 2 
2—Bouche pourvue de lévres ou dents...............--2-00: 3 
PAR te CMMI EES eco: ova si és 2s. sb aig ee bee ee ee eme 5 
3—Bouche a six petites lévres papilliféres................... Serticeps 
Bauckera ers Sevres Ol dents... 2... .. 0.0 eens oaseee ene 4 
A— Tete ormee de lobes cuticulaires. ©. o.006cc6.0. oes Seem cea ck Schistorophus 
STE: TT. 4 ae odo Re Oe GErE eae ee teeeirmne aon a Rhabdochona 
5 Malesiaranes-candales (bourse) ...%.. 6.5.2. idessee. sexe es 6 
Males sans bourse; courte capsule buccale................ i 
6—Vestibule allongé; papilles préanales couplées............ Cystidicola 
Courte capsule buccale; papilles préanales simples.........Ceratospira 
7—Queue obtuse, arrondie; vulve antérieure................. Thelazia 
Queue pointue, oxyuriforme; vulve postérieure........... Oxyspirura 


Genre Thelazia Bosc, 1819 (Thelazius Bosc, 1819; Thalazia de 
Blainville, 1819).—Bouche sans lévres, suivie d’une capsule buccale; 
bord antérieur de la capsule retroussé en dehors et découpé en six 
festons par des échancrures dont quatre paraissent occupées par un 
petit organe papilliforme trés réfringent. Deux papilles céphaliques 
latérales et quatre submédianes. 

Male a queue obtuse ordinairement recourbée en crochet, sans ailes 
latérales; un grand nombre de papilles préanales dont une médiane, 
impaire, au-dessus du cloaque; trois ou quatre (?) papilles postanales. 
Deux spicules inégaux. 

Femelle a queue conique mousse, arrondie, portant deux papilles 
laterales a son extremité. Vulve située antérieurement, un peu en 
arriere de la terminaison de l’oesophage; deux branches utérines 
dirigées en arriere. Embryons éclosant dans les utérus. 

Habitat.—L’habitat normal est représenté par les canaux exréteurs 
des glandes lacrymales des mammiféres, d’ot les Vers s’échappent 
assez souvent pour glisser sous les paupiéres ou a la surface de l’oeil; 
on en a signalé exceptionnellement a l’intérieur du globe oculaire. 
Certaines formes semblent se rencontrer sous la membrane nictitante 
des Oiseaux. 

Espece type: Thelazius Rhodesii Desmarest, 1827. 

A.—Especes des Mammiferes. 

1—Thelazia rhodesi (Desmarest, 1827).—Syn.: Thélazie de 
Rhodes Bose, 1819; Thelazius Rhodesii Desmarest, 1827; Thelazia 
Rhodesui de Blainv., 1828; Filaria bovis Baillet, 1858; Filaria palpe- 
brarum Baillet, 1858; “Filaria lacrymalis Gurlt” Baillet, 1866 et 
Railliet, 1893, pro parte-——Chez le Boeuf (Bos taurus) et le Buffle 
(Buffelus bubalis). 


RAILLIET—LA FAMILLE DES THELAZIIDAE 101 


2.—Thelazia gulosa Railliet et Henry, 1910.—Chez le Bos taurus. 

3.—Thelazia alfortensis Railliet et Henry, 1910—Chez le Bos 
taurus. 

4. Thelazia leesei Railliet et Henry, 1910—Chez le Camelus 
dromedarius, dans ’humeur vitrée, dans un kyste du corps clignotant ; 
commune sous les paupieres, principalement sous le corps clignotant 
et dans le conduit de la glande de Harder. 

5.—Thelazia lacrymalis (Gurlt, 1831)—Syn.: Filaria lacrymalis 
Gurlt, 1831, pro part e; Filaria palpebralis Wilson, 1844, non Pace, 
1867; “Filaria palpebralis Wilson” Railliet, 1893.—Chez le cheval 
(Equus caballus). Aurait été trouvée par Busch dans l’humeur 
aqueuse du meme animal. 

6.—Thelazia callipaeda Railliet et Henry, 1910.—Chez le Canis 
familiaris. Parait commune en Birmanie. 

B.—Espéces parasites des Oiseaux.—Ne possedent pas la papille 
impaire précloacale. 

Thelazia anolabiata (Molin, 1860).—Syn.: Spiroptera anolabiata 
Molin, 1860; Filaria anolabiata Stossich, 1897; Oxyspirura ? anoliabi- 
ata Ransom, 1904.—Chez le Crax fasciata, sous la nictitante et a la 
surface de l’oeil. 

Thelazia papillosa (Molin, 1860)—Syn.: Spiroptera papillosa 
Molin, 1860; Oxryspirura ? papillosa Ransom, 1904.—Chez Thrasactus 
harpyia et Geranospizias caerulescens, sous la nictitante. 

Thelazia campanulata (Molin, 1858)—Syn.: Filaria campanulata 
Molin, 1858.—Chez Rupornis magnirostris, sous la nictitante. 

Thelazia ? cirrura (Leidy, 1886).—Syn.: Filaria cirrura Leidy, 
1886.—Chez Megaquiscalus major, dans Vorbite. 

Thelazia ? stereura (Rud., 1819).—Syn.: Spirotera stereura Rud., 
1819; Oxyspirura ? stereura Ransom, 1904.—Chez Aquila maculata, 
sous la nictitante et dans le méat auditif. 


Genre Ceratospira Schneider, 1866.—Teéete nue. Bouche entourée 
de papilles et suivie d’une courte capsule buccale. 

Males a queue trés courte, mousse, pourvue de larges ailes; de 
chaque coté une rangée longitudinale de papilles simples, dont 9 a II 
préanales. 2 spicules tres inégaux, 

Femelles a queue trés courte, mousse. Vulve trés antérieure. Par- 
fois vivipares. 

Habitat—Cavité orbitaire des Oiseaux. 

Espeéce type. C. ve siculosa Schneider, 1866. 

1.—Ceratospira ve siculosa Schneider, 1866.—Cavité orbitaire de 
PEclectus pectoralis. 

2.—Ceratospira ophthalmica (Linstow, 1898)—Syn.: Ancyracan- 
thus ophthalmicus Linstow, 1898; Ceratospira ophthalmica Ransom, 
1904.—Cavité orbitaire du Zonoenas brenchleyi. 


102 THE JOURNAL OF PARASITOLOGY 


Genre Schistorophus n.g. (Tetracanthus Hemprich et Ehrenberg, 
1866, non Hope, 1835; Ancyracanthus Schneider, 1866, pro parte, non 
Diesing, 1838).—Téete ornée de quatre lobes cuticulaires aigus, con- 
fondus en avant avec la cuticule, plus ou moins réunis a leur origine, 
surtout sur les lignes médianes, et disposés en toit. Bouche petite, 
géenéralement a deux petites levres ou dents. Un vestibule allongé. 
(Esophage composé de deux parties. 

Males a queue mousse, arrondie, pourvue d’ailes latérales et de 
nombreuses papilles, les préanales disposées de chaque cdté en une 
longue série simple. Deux spicules inégaux. 

Femelles a queue courte, conique, plus ou moins obtuse; vulve dans 
la région postérieure ou moyenne du corps. Parfois vivipares. 

Habitat—Emtre les tuniques du gésier des Oiseaux. 

Espece type: Ancyracanthus longicornis Hemprich et Ehrenberg, 
1866. 

1—Schistorophus longicornis (Hemprich et Ehrenberg, 1866.— 
Syn.: Ancyracanthus longicornis Hemprich et Ehrenberg, 1866.— 
Entre les tuniques du gésier de Numenius arquatus, Tringa variabilis, 
Totanus glottis. 

2.—Schistorophus bicuspis (Rud., 1819).—Syn.: Spiroptera bicus- 
pis Rud., 1819; Dispharagus bicuspis. Duj., 1845; Histiocephalus 
gracilis Dies., 1851; Histiocephalus bicuspis Linstow, 1878.—Entre les 
tuniques du gésier de Squatarola helvetica. Probablement identique a 
la forme précédente. 

3.—Schistorophus bidens (Rud., 1819).—Syn.: Spiroptera bidens 
Rud., 1819; Dispharagus bidens Duj., 1845; Spiroptera denticulata 
Molin, 1860; Ancyracanthus bidens Schneider, 1866—Entre les 
tuniques du gésier de Merops apiaster et peut-etre d’Astur palumbarius. 

4.—Schistorophus laciniatus (Molin, 1860).—Syn.: Histiocephalus 
laciniatus Molin, 1860.—Entre les tuniques du gésier de Rallus 
cayennensis. 

5.—Schistorophus (?) umbellifer (Molin, 1860).—Syn.: Spiroptera 
umobellifera Molin, 1860.—Entre les tuniques du gésier d’ibis rubra et 
de Totanus melanoleucus. 

6.—Schistorophus (?) spinulosus (Molin, 1860).—Syn.: Filaria 
spinulosa Molin, 1860.—Entre les tuniques du gésier de Glareola 
austriaca. 

7.—Schistorophus (?) acanthocephalicus (Molin, 1860).—Syn.: 
? Strongylus ambiguus Rud., 1802; ? Spiroptera Sternae Rud., 1819; 
? Spiroptera sternae hirundinis Deslongchamps, 1824; Spiroptera 
acanthocephalica Molin, 1860.—Entre les tuniques du gésier de Sterna 
caspica; peut-étre dans l’oesophage de Sterna hirundo. 


RAILLIET—LA FAMILLE DES THELAZIIDAE 103 


8.—Schistorophus (?) capillaris (Molin, 1860).—Syn.: Spiroptera 
capillaris Molin, 1860; Cheilospirura capillaris Diesing, 1861.—Entre 
les tuniques du gésier de Sterna hirundo. 


Genre Serticeps n.g—Tete ornée d’appendices ou festons multiples 
et variés. Bouche a six petites levres portant chacune une petite 
papille. 

Males a queue obtuse; ailes caudales asymétriques; 10 paires de 
papilles préanales. Deux spicules trés inégaux. 

Femelles a queue obtuse. Vulve voisine de l’anus. 

Habitat——Entre les tuniques du gésier des Oiseaux. 

Espéce type: Spiroptera vulvoinflata Molin, 1860. 

1.—Serticeps vulvoinflatus (Molin, 1860.)—Syn.: Spiroptera vul- 
voinflata Molin, 1860.—Entre les tuniques du gésier de Tyrochilus 
ochropygus. 


Genre Cystidicola Fischer de Waldheim, 1897—Téte nue. Bouche 
circulaire suivie d’un vestibule cylindrique. Césophage tres long. 

Males a queue arrondie a l’extrémité; ailes caudales minces; de 
chaque coté, une longue rangée de papilles préanales couplées et de 
papilles postanales simples. Deux spicules inégaux. 

Femelles a queue droite, mousse. Vulve dans la région moyenne 
ou antérieure du corps; utérus opposés. CMEufs nombreux, a coque 
épaisse, pourvus, au moins dans le type, de filaments polaires. 

Habitat.—Vessie aérienne, plus rarement cesophage, des Poissons 
d’eau douce. 

Espéce type: Cystidicola farionis Fischer, 1797. 

1—Cystidicola farionis Fischer, 1797.—Syn.: Fissula cystidicola 
Lamarck, 1800; Ophiostoma cystidicola Rud., 1801; Spiroptera cystidi- 
cola Rud., 1819; Dispharagus cystidicola Duj., 1845; Ancyracanthus 
cystidicola Schneider, 1866.—Vessie aérienne de Trutta fario, Tr. 
trutta, Squalius cephalus; vessie aérienne et cesophage de Thymallus 
vulgaris; cesophage de Coregonus oxyrhynchus. 

2.—Cystidicola impar (Schneider, 1866).—Syn.: “Gordius argil- 
laceus L.” Martin, 1771; Ancyracanthus impar Schneider, 1866.— 
Vessie aérienne d’Osmerus eperlanus, Gasterosteus aculeatus, Trutta 
fario, Coregonus albula, C. fera, C. lavaretus. 

3.—Cystidicola (?) serrata (Wright, 1879) —Syn.: Ancyracanthus 
serratus R. Wright, 1879—Coeur de Coregonus albus. 


Genre Galeiceps n.g—Téte pourvue d’un renflement qui la coiffe 
a la facon d’un couvercle ou d’un casque. Bouche a quatre bourrelets 
séparés sur la surface ventrale et portant chacun a son bord interne 
une dent conique. 


104 THE JOURNAL OF PARASITOLOGY 


Males a queue obtuse ; nombreuses papilles préanales simples. Deux 
spicules égaux. 

Femelles a queue trés courte et pointue. 

Habitat.—Intestin des Marsupiaux. 

Espéce type: Ancyracanthus cucullus Linstow, 1899. 

1—Galeiceps cucullus (Linstow, 1899).—Syn.: Ancyracanthus 
cucullus Linstow, 1899.—Intestin de Potamogale velox. 


Genre Rhabdochona n.g—Tete nue. Bouche a deux levres limitant 
une cavité infundibuliforme soutenue par des batonnets longitudinaux. 
(Esophage de médiocre longueur, composé de deux parties distinctes. 

Males a queue conique, pointue, recourbée; pas d’ailes caudales; 
nombreuses papilles préanales et postanales simples. Deux spicules 
inégaux. oe 

Femelles a queue droite, conique, allongée. Vulve vers le tiers 
postérieur du corps; utérus opposés. 

Habitat.—Intestin des Poissons d’eau douce. 

Espece type: Dispharagus denudatus Duj., 1845. 

1—Rhabdochona denudata (Duj., 1845).—Syn.: ? Fusaria cunei- 
formis Zeder, 1800; ? Ascaris cuneiformis Rud., 1809; Dispharagus 
denudatus Duj., 1845; Histiocephalus denudatus Dies., 1851; Cucul- 
lanus pachystomus Linstow, 1873; ? Dispharagus filiformis Zschokke, 
1884; Ancyracanthus denudatus Linstow, 1887; Ancyracanthus denu- 
datus Linstow, 1902.—Intestin de nombreaux Cyprinidés. 


Genre Oxyspirura Drasche, 1897.—Tete nue, rarement avec un 
renflement cuticulaire. Bouche sans lévres, suivie d’une courte capsule 
buccale. Queue trés aigiie, oxyuriforme. 

Males a queue généralement incurvée ou spiralée, dé pourvue 
d’ailes latérales; papilles non pédonculées, les préanales en nombre 
assez variable (2 a 28), les postanales (1 a 8) souvent asymétriques. 
Deux spicules tres inégaux. 

Femelles a queue droite. Vulvedans la partie postérieure du corps, 
un peu en avant de l’anus. 

Habitat.—Sous la nictitante des Oiseaux. 

Espece type: Spiroptera cephaloptera Molin, 1860. 

1—O-xyspirura cephaloptera (Molin, 1860).—Syn.: Spiroptera 
cephaloptera Molin, 1860; Cheilospirura cephaloptera Diesing, 1861; 
Oxyspirura cephaloptera Stossich, 1897—Sous la _ nictitante de 
Momotus momata et d’Icterus croconotus. 

2.—O-xyspirura anacanthura (Molin, 1860).—Syn.: Spiroptera 
anacanthura Molin, 1860; Oxyspirura anacanthura Stossich, 1897— 
Sous la nictitante de Crotophaga ani et Cr. major. 


RAILLIET—LA FAMJLLE DES THELAZIIDAE 105 


3.—Ov-xyspirura brevisubulata (Molin, 1860)—Syn.: Spiroptera 
brevisubulata Molin, 1860 ,Oxyspirura brevisubulata Stossich, 1897.— 
Sous la nictitante d’Otus choliba. 

4.—O-xyspirura mansoni (Cobbold, 1879).—Syn.: Filaria Mansoni 
Cobbold, 1879, non Zune, 1892; Spiroptera Emmerezi Mégnin, 1901; 
Spiroptera Mansoni Marotel, 1903; Oxyspirura Mansoni Ransom, 
1904.—Sous la nictitante de Gallus domesticus, Meleagris gallopavo, 
Pavo cristatus domesticus. 

5.—Oxyspirura parvovum G. Sweet, 1910.—Syn.: O-syspirura 
parovum Breinl., Taylor et Johnston, 1913.—Sous la nictitante et dans 
la fosse lacrymo-nasale de Gallus domesticus. 

6.—Oxyspirura ophthalmica (Linstow, 1903)—Syn.: Cheilospi- 
rura ophthalmica Linstow, 1903; Osyspirura ophthalmica Ransom, 
1904.—CEil de Turnix taigoor. 

7—Oxyspirura siamensis (Linstow, 1903).—Syn.: Cheilospirura 
siamensis Linstow, 1903; Ovyspirura siamensis Ransom, 1904.—Chez 
Centropus sinensis (probablement oeil). 

8. Oxyspirura anthochoerae Johnston, 1911—Syn.: Ascaris sp. 
Krefft, 1873; Ceratospira anthochoerae Johnston, 1911; Oxyspirura 
anthochoerae Johnston, 1912—QEil Anthochoera caruncalata. 

On a fait en outre rentrer dans ce genre diverses autres formes 
au sujet desquelles les plus grandes réserves s’imposent, par exemple: 

Spiroptera sigmoidea Molin, 1860, cavité orbitaire de Corvus 
frugilegus. 

Spiroptera brevipenis Molin, 1860, sous la nictitante de Cariama 
cristata. 

Spiroptera heteroclita Molin, 1860, sous la nicitante de Nothocrax 
urumutum, 

Spiroptera acuminata Molin, 1860, intestin de Brycon falcatus. 

Spiroptera spiralis Molin, 1860, sous la plante des pieds des 
Edentés: Bradypus cuculliger et Choloepus didactylus. 


SEASONAL DISTRIBUTION OF SOME ACANTHOCEPHALA 
FROM FRESH-WATER HOSTS * 


H. J. VANCLEAVE 


In a recent paper Linton (1914: 48-56) has given a brief survey 
of the evidence on seasonal distribution of parasites of marine fishes. 
He concluded this paper with the statement: “There does not appear 
to be evidence of any marked periodicity in the occurrence of helminth 
parasites of marine fishes, either adult in the alimentary canal, or 
immature encysted in the tissues of their hosts, beyond what may be 
expected where fishes are exposed to varying sources of infection in 
the course of their migrations.” In speaking of the seasonal distribu- 
tion of Acanthocephala he has recorded the occurrence of Echinorhyn- 
chus gadi Muller (which he called E. acus) in Pseudopleuronectes 


‘ 


americanus *. in every month in which examinations were made, 
viz., January, February, April, May, July, August, September, 
October, November, and December.” The mere fact that a parasite 
is present in its final host for the greater part of, or even for the entire, 
year is not proof that there is no periodicity in its occurrence. One 
generation of parasites might overlap another generation, yet if con- 
ditions for reinfestation were such that larvae could enter the final 
host only at restricted periods it would be possible to detect a periodic- 
ity in the infestation upon the basis of the distinctions between imma- 
ture and mature individuals. Unfortunately most records make no 
mention of age of the parasites. On the other hand, if the intermediate 
host of the parasite constitutes a part of the food of the final host 
throughout the year the chances for constant reinfestation make it 
impossible to recognize distinct cycles of infestation. 

Conditions of life in fresh-water are so much more varied than 
in the ocean that it would not be surprising to find seasonal changes 
in kinds of parasites and degrees of infestation more marked in hosts 
from fresh water than in hosts from the ocean. Very little has been 
done toward establishing any correlation between extent or degree of 
parasitic infestation and periodicity of occurrence. The records on . 
these topics deal almost exclusively with the general problem of the 
number of parasites found in a given host without further analysis 
beyond an occasional tabulation of the data for the classes or orders 
of the parasites found. A number of writers have recorded the 


* Contributions from the Zoological Laboratory of the University of Illinois, 
No. 58. 


VAN CLEAVE—DISTRIBUTION OF ACANTHOCEPHALA 107 


months in which they have found Acanthocephala in various hosts 
without furnishing any data on the presence or absence of the same 
parasites in the same hosts at other times of the year. Thus Zschokke 
(1884: 58) has recorded Pomphorhynchus laevis (Muller) from vari- 
ous fishes from January to June, but he has not given any evidence 
or proof of its absence for the remainder of the year. In fact his 
records seem only to indicate the dates when he chanced to examine 
fish which were infested with P. laevis rather than to represent an 
attempt on his part to establish the limits of the seasons when this 
parasite occurs in its final host. ; 

In collecting fresh-water Acanthocephala the writer has been 
impressed by the varying degrees of infestation of certain hosts at dif- 
ferent times of the year. A search of the literature has furnished so 
little actual information upon this subject that it seemed worth while 
to investigate the question, especially since Linton has rather sum- 
marily dismissed the topic with a brief generalization. The records 
from which the following data have been gathered comprise three 
species of Acanthocephala. In one of these no marked seasonal distri- 
bution is evident, while in the other two definite seasonal cycles mark 
the occurrence of the parasite in its final host. 


Neoechinorhynchus* emydis (Leidy) occurs in the intestine of 
a number of fresh-water turtles. The records of the writer and of 
Mr. H. W. Stunkard include the examination of over 200 individuals 
belonging to species susceptible to infestation with N. emydis. These 
came from Iowa, Ohio, West Virginia, Texas, and various points in 
Illinois. The unselected data from all the records when assembled 
and tabulated presented evidence of a seasonal distribution of N. 
emydis, which upon closer examination of the data proved to be spurt- 
ous. Parasites of this species were recorded from hosts examined in 
October, November, December, January, and February with a few 
records of occurrence in July. Records of examinations in April, June, 
and September showed no infestation with this parasite. This in itself 
seemed to indicate a restriction of N. emydis in the intestine of its 
final host to a limited portion of the year. However, further examina- 
tion revealed that certain localities within the geographical range of 
the species are free from that parasite. By a strange coincidence 
turtles happened to be examined from these localities in months when 
no records were available for regions where N. emydis is known to 
occur. This shows how statistical data if not carefully checked may 
give false evidence of cyclic occurrence of an organism. 


* Neoechinorhynchus, Stiles and Hassall, 1905=Eorhynchus, VanC., 1914= 
Neorhynchus, Hamann, 1892, preoccupied. 


108 THE JOURNAL OF PARASITOLOGY 


Indirect evidence has shown that N. emydis occurs in the intestine 
of its final host throughout the year. Turtles from Havana, Ill., have 
been kept without food in aquaria fed by the University of Illinois 
water supply, which is from deep wells, for about eleven months. At 
the end of this time one turtle still harbored twelve mature specimens 
of N. emydis in its intestine. The evidence of an original infestation 
lasting practically a year, together with the fact that in many instances 
the writer has found fully mature, immature, and intermediate speci- 
mens in the intestine of the same individual proves that turtles in 
regions where N. emydis occurs are constantly exposed to reinfesta- 
tion with that species. Consequently there is no cyclic change in the 
degree of infestation from month to month. 

It is interesting to note that while N. emydis has a broad geogra- 
phical distribution, occuring in the records under consideration at 
certain points in Illinois, North Carolina, and Texas, it is by no means 
generally distributed over its range. In Illinois, for example, turtles 
of species susceptible to infestation with N. emydis have been collected 
at Urbana, Muncie, and Chicago, and in no case has a single specimen 
of N. emvdis been found. It seems strange that a species with such a 
broad dispersal should not have followed the dispersal of its final host. 
This probably finds explanation upon the grounds that in the localities 
where the parasite does not now occur if it was originally or subse- 
quently introduced the embryos when expelled from the intestine of 
the final host were not taken up by animals in which the larvae could 
develop or in case they did find lodging in a host it must have been in 
some animal which was not used by the turtles as food. Thus through 
the lack ef adaptability to new conditions brought on by the specializa- 
tion accompanying parasitism this species has been excluded from some 
regions which are included within its limits of distribution. 

In contrast with the lack of periodicity in the species just discussed 
may be noted the condition found in Neoechinorhynchus gracilisentis 
(VanC.) found in the intestine and intestinal caeca of the gizzard-shad, 
Dorosoma cepedianum (LeSueur), from the Illinois River system. 
During the period from 1909 to 1912 the writer examined more than 
300 gizzard-shad for parasites. But two species of parasites have been 
found. Both of these were Acanthocephala belonging to the genus 
Neoechinorhynchus. N. gracilisentis has been found in October, 
November, December, February, March, and April, but specimens at 
these different dates displayed wide variation in degree of sexual 
maturity. Those collected in October were almost invariably small 
and immature, with a high percentage of infestation. By the latter 
part of November individuals of this species had reached full sexual! 
maturity, as indicated by the numbers of hard-shelled embryos con- 
tained in the body cavities of the females. In April the percentage 


VAN CLEAVE—DISTRIBUTION OF ACANTHOCEPHALA 109 


of infestation had decreased to less than one half of that found for 
October, and the number*of individuals per host also had decreased 
though every parasite had reached full sexual maturity and the maxi- 
mum size for the species. Numerous examinations in the months of 
June, July, and August have failed to give even a single specimen of 
this species. From the foregoing data it is evident that the introduc- 
tion of N. gracilisentis into the final host must occur in early fall, 
probably in September. The individuals have become fully mature 
by April and disappear entirely from the final host during the months 
of June, July, and August. In an earlier paper (VanCleave 1913; 181) 
I have indicated the probable relationship between this periodicity of 
infestation and the food habits of the gizzard-shad. Observations 
upon the stomach contents of the shad, which is primarily a scavenger, 
have failed to throw any light upon the probable intermediate host 
of this parasite. The entire digestive tract is usually filled with mud 
and decomposed plant tissues with a very few shelled rhizopods and 
some species of microcrustacea. 


TABLE SHOWING SEASONAL DISTRIBUTION OF THREE FRESH-WATER SPECIES 
OF NEOECHINORHYNCHUS 


| 
| 
> 5 sang, 
Species ue ae euikte hoe | epee 
me a= ea he Hes = ie a a ee 
aetes cine Rectorate tea |e | -8 | Bo) g 
Se Le aust | a | oe |} oO] a] 6 
_ a 
aes | | 
ING ORLY CUS vceiacnxwees os +] + x > Sell Rib Sarg NS fe ya. 4 pe = a | a 
| | | | 
N. gracilisentis......... x a + +/+/]/—|]—-|/-|/2+ ae =e | Hs 
N. longirostris.......... —;}/—]—};—]+ + + | + x x i fe 


* For additional experimental evidence see text. 

+ Positive records of occurrence based upon examination of hosts. 

— Absence from all hosts examined. 

+ Extremely probable occurrence. Though definite records of infestation are wanting 
the stage of maturity of individuals collected the preceding or the following month indicates 
that a complete gradation of stages in development necessitates an overlapping of infes- 
tation into adjacent month. 

X records, both positive and negative, wanting though stages of maturity of the 
parasites in the two adjacent months together with the data upon longevity of the species 
in the final host justifies the assumption of a positive infestation. 


Neoechmorhynchus longirostris (VanC.), the second species found 
in the intestine and intestinal caeca of the shad, occurs in much smaller 
numbers and in but a very small percentage of fishes examined. Imma- 
ture individuals were found in June and July. Gravid females were 
found in August, November, and December. While the number of 
records is insufficient to permit of establishing all points in a seasonal 
cycle, yet the evidence at hand indicates that the host is free from 
parasites of this species from late winter until early summer. 

In the case of both species of Neoechinorhynchus from the gizzard- 
shad the relatively short life in the body of the final host is noticeable. 


110 THE JOURNAL OF PARASITOLOGL 


Moreover, the parasites of a given species collected at the same time 
from a given region have all reached approximately the same stage in 
development. This indicates that the period when infestation may 
occur is very brief. Attention should also be called to the fact that 
periods of infestation in these two species are not coexistent. 


CONCLUSIONS 


1. Seasonal distribution of fresh-water Acanthocephala varies in 
different species. No general statement can be made to apply to the 
entire group. 

2. Neoechinorhynchus emydis (Leidy) has broad limits of geo- 
graphical distribution, but has never been found in turtles of suscepti- 
ble species from some localities within its range of distribution. 

3. N. emydis occurs in turtles from some localities at all seasons of 
the year. 

4. The same host may harbor specimens of N. emydis in all stages 
of development between immature and fully mature. This shows the 
host must be exposed constantly to sources of infestation. 

5. There is no cyclic change in the degree of infestation with this 
species from month to month. 

6. N. gracilisentis (VanC.) enters the gizzard-shad in early fall, 
probably September; in April or May it attains sexual maturity and 
is finally expelled. During the summer the gizzard-shad is not para- 
sitized by this species. 

7. N. longirostris (Van.C.) parasitizes the gizzard-shad in the 
summer, reaches full sexual maturity by midwinter, and disappears 
entirely from spring to early summer. 

8. The demonstrable presence of a seasonal cycle in the life history 
of a parasite involving two or more hosts is dependent upon (a) 
longevity of the parasite in the final host; (b) extent of the time in 
which infestation of the final host may occur; (c) length of time 
required for development of the larva in the intermediate host; (d) 
seasonal changes in the food habits of the final host, or active migra- 
tions of the host to and from sources of infestation. 


LITERATURE CITED 


Linton, Edwin. 1914. On the Seasonal Distribution of Fish Parasites. Tr. 
Amer. Fish. Soc., 44: 48-56. 

Stiles, C. W., and Hassall, A. 1905. The Determination of Generic Types, 
and a List of Roundworm Genera, with Their Original and Type Species. Bull. 
Bur. An. Ind., 79: 150 pp. 

VanCleave, H. J. 1913. The Genus Neorhynchus in North America. Zool. 
Anz., 43: 177-190. 

Zschokke, Fr. 1884. Recherches sur l’organisation et la distribution zoo- 
logique des vers parasites des poissons d’eau douce. Thesis Genéve; 89 pp. 


ON THE INTERMEDIATE HOSTS OF THE LUNG DISTOME, 
P. WESTERMANI KERBERT 


Sapao YOSHIDA 
Pathological Department of Osaka Medical University 


Japan is famous for being considerably infested with lung distome. 
lt is about thirty-four years since the human lung distome was dis- 
covered by Kiyono, Yamagata, Nakahama, and Suga in Okayama 
prefecture, near the center of the country. During these years though 
numerous reports on symptoms of patients and on diagnosis and 
many pathological notes have been published by investigators from 
various districts of the country, the result of the developmental investi- 
gations has not yet been achieved, nothing being known of a life his- 
tory of the parasite. Fortunately, however, light has been thrown 
upon the subject by the recent discovery of an intermediate, probably 
the second, host of this worm by Koan Nakagawa, director of the 
Shinchiku Hospital, Formosa, who has been earnestly investigating 
this parasite in Formosa for several years. 

I have also studied experimentally the life-history of this worm, 
and found certain species of fresh water crabs as intermediate hosts. 
I shall here communicate the results obtained from my experiments. 


INTERMEDIATE HOST 


In Formosa Nakagawa found the encysted larvae in two fresh 
water crabs and experimentally proved that they grew up to the lung 
distomes. The two crabs were identified by A. Terao as follows: 
Potanon (Geothelphusa) obtusipes (Stimpson), P. (Ge.) dehaanii 
(White). 

Nakagawa added that a fresh-water crab Eriocheir japonicus (De 
Haan) will also probably prove to be the intermediate host. 

I have experimentally proved that the encysted larvae of this worm 
are found in three species of fresh-water crabs from various districts 
of Japan proper. They are identified as follows: 1. Patamon dehaanii 
(White) ; 2. Sesarma dehaani (Milne Edwards) ; 3. Eriocheir japonicus 
(De Haan). 

These species can be found in any part of Japan. P. dehaanii ts 
small in size and light reddish brown or purple in color. It is a com- 
mon crab in the shallow water of a mountain stream. This crab 1s 
edible and used for food commonly in some districts and rarely in 
other parts of this county. It can be eaten raw or cooked. FE. japan- 


112 THE JOURNAL OF PARASITOLOGY 


icus is large in size and dark brown or black in color, very common 
in any brook and river of Japan, including Formosa and Corea. 
Large hairy forceps are characteristic of this species. It also is edible 
and commonly used for food in all districts, though generally eaten 
cooked being roasted, boiled, or fried. S. dehaani is of median size, 
the same in color as E. japonicus, having light reddish purple forceps. 
It lives generally in the lower parts of a river in various parts of our 
country. This species is not used for food. 

Distribution of Encysted Larvae in Body of Intermediate Hosts — 
The encysted larvae occur generally in the liver, muscles, and gills of 
the host. The distribution of the encysted larvae varies but slightly 
according to the species of the host, so far as | have examined. In 
P. dehaanti and P. obtusipes, they are found frequently in the liver, 
and rarely in muscles and gills; in E. japonicus chiefly in gills, muscles, 
and hypodermis, and rarely in the liver; and in S. dehaant mainly in 
the liver and very rarely in the gills. In the liver the encysted larvae 
are attached loosely to the lobes of the organ so that they may be 
easily detached. In the gills they adhere between the lamellae in the 
case of P. dehaanu, but are found only in the blood vessel running 
through the median line of the upper surface of the gill in the case 
of japonicus. In the latter species they occur not only in the muscles 
of the trunk, but in muscle and hypodermis of all appendages. The 
encysted larvae in the muscles and hypodermis or in the blood vessels 
are easily movable. 

Frequency of Occurrence and Number of Encysted Larvae in Host. 
—It is reported by Nakagawa that about 100 per cent. of P. obtusipes 
are infected with the encysted larvae at Shinchiku, the most famous 
district for the lung distome in Formosa. Ryo Ando reported that 
about 40 to 70 per cent. of P. dehaanu of Gifu prefecture are infected 
with the larvae. According to my own examination E. japonicus of 
Tokushima and Okayama prefectures is infected to the extent of 
about 70 to 85 per cent., and S. dehaani of Osaka prefecture to about 
20 to 80 per cent. The number of the cysts in one crab varies consid- 
erably according to species of the host, and even in the same species 
it varies according to locality and other conditions. In my own exami- 
nations I found some F. japonicus infected with several hundreds of 
the encysted larvae while others were infected only with a few in 
spite of the locality being the same. S. dehaani was generally infected 
with 2 to 30 cysts of the larvae. In P. dehaanii from Okayama and 
Nigigsta prefectures I found only a few cysts while Ando is reported 
to have obtained several hundreds of cysts in the same species from 
Gifu prefecture. 


YOSHIDA—INTERMEDIATE HOSTS OF LUNG DISTOME 113 


I give here tables showing the percentages of the infected crabs 
and the numbers of the cysts in the hosts examined by myself. 


TABLE 1.—S. DEHAANI FROM HIEJIMA, OSAKA PREFECTURE 


| | 
| Number | Number! Percentage | Maxi- | Mini- | Total | Average 
of of Crabs) of mum mum Number’! Number 
Date Crabs |Infected| Infected | of Cysts of Oysts of Cysts of Cysts | Number 
xam- with Crabs in One | inOne inCrabs| in One | 
ined Cysts | Crab Crab Infected| Crab 
10/VI 19 4 | 21.05% 3 L 7 1.43 1 
NVI 8 2 | 25.00% 2 2 4 2.00 2 
12/VI 3 1 33.33% | 2 2 2 2.00 3 
14/VI 20 2 10.00% 2 ik 3 1.50 4 
15/VI 12 2 16.66% 3 1 4 2.00 5 
16/VI 12 1 8.33% 1 iz | if | 1.00 6 
17/VI 21 5 23.80% z. Z Gray 120 vt 
18/VI 8 3 37.50% 4 1 7 2.33 8 
22/VI | 24 2 8.33% 2 1 3 1.50 9 
28/VI 9 2 22.22% 3 2 5 2.50 10 
10/VII 10 2 20.00% | 8 Oe | 816 5.00 11 
11/VII 9 % 22.29% =| 7 3 10 5.00 12 
12/VII 1 L 100.00% 1 1 1 1.00 13 
13/VIL l1 3 27.27% 3 1 6 2.00 14 
14/VII_ | 9 tf 44.44% 4 ul 10 2.50 15 
15/ VII 16 2 12.50% 2 1 3 | 1.50 16 
16/VIL 8 2 | 25.00% 3 2 5 | 2.50 17 
200 | 40 | 20.00% 8 1 87 2.17 17 
TABLE 2.—S. DEHAANI FROM EBIE, OZAKA PREFECTURE 
| Number wamben| Percentage | Maxi- Mini- Total Average | 
of of Crabs of mum mum Number | Number | 
Date _ Crabs Infected, Infected | of Cysts, of Cysts| of Cysts| of Cysts Number 
| Exam- with Crabs in One | inOne |in Crabs} in One | 
ined Cysts | Crab Crab  Infected| Crab 
28/ VIII 5 4 | 80.0% 29 9 89 22.5 1 
30/ VIII 2 2 | 100.0% 26 26 52 26.0 Pe 
1/1X 6 | 5 | 83.3% 30 15 107 21.4 3 
2/1X 5 5 100.0% | 22 ib 49 9.8 a 
18 16 | 888% 30 1 297 18.5 4 
1 


TABLE 3—E. JAPONICUS FROM IKUINA, TOKUSHIMA PREFECTURE 


23/VII 
25/VIL 
26/VIL 
27/VIL 
28/VII 
2/VIII 
3/VIIL 
3/VII1 
6/VIIL 
7/VI1 


| Number Number | Percentage Maxi- Mini- | Total Average | 
of of Crabs | of mum | mum | Number) Number | 
| Crabs | Infected Infected of Cysts | of Cysts | of Cysts| of Cysts Number 
| Exam- | with | Crabs in One | inOne |inCrabs| in One 
_ ined Cysts | Crab | Crab | Infected} Crab 
10 9 90 % 19 3 96 10.66 i 
4 4 100 % 26 CO if 56 14.00 2 
7 7 100 % 14 | 1 29 4.14 3 
eine 4 ae 86.6% 2 1 6. | 4.69 4 
| 1 1 100 % 86 86 86 86.00 5 
10 3 33.38% 77 7 161 59.66 6 
15 7 46.6% 40 | ik 7 10.00 7 
5 3 60.0% win OS we Ag ALE 8 
8 6 75 % 48 2 85 14.16 9 
2 if 50 % 343 343 | 843 343.00 10 
77 54 70.1% 343 | 1 987 19.35 10 


TABLE 4.—DISTRIBUTION OF CYSTS IN THE BODY OF NO. 10 IN TABLE 3 


CUB OME HOLY RIOER cote. csatsisl- ceiss.oteislelo ee samaioe nie Sia acto e = voce cscs eas 
Body muscles on left side 
Body muscles on right side 
Forceps on right side 
Third leg on right side 
Third leg on left side 


&1 


114 


TABLE 5.—DISTRIBUTION OF CYSTS 


PRECEDING TABLE 


. THE JOURNAL OF PARASITOLOGY 


IN THE LEGS OF THE CRAB IN THE 


ooo eee 


Ischiopodite | Carpop- Propo- Dactylop- Total 
Meropodite odite dite odite 
Bight fOrcaphe.sc. sc sce0 se 4 12 3 0 19 
Right third leg.............| 7 | 3 2 0 12 
ISTE BIPU OR ases sakatvenc's } 13 6 4 0 23 
POUH hanes cece cence 24 21 9 0 54 
| | 


TABLE 6.—NINE CYSTS IN GILLS, FIFTY-FIVE IN MUSCLES AND HYPODERMIS 


| 

First Legs | Second Third Fourth Fifth 
(Forceps) | Legs Legs Legs Legs 

Right} Left Right! Left Right | Left Right Left | Right; Left 
Muscles attached.......... 1 2 | 2 tt G's ta iene 2 1 1 
IBASIDOGILO. 6 cc.ssc< c.cisz so ss iL Ou, «2 1 0 7 aie a Al! 0 0 
Ischiopodite..............- a 3 0 2 0 2 1 0 0 2 
IMGTODOGItG: ses icjos 05 = <5 4 3 0 i) 2 0 0 2 1 1 
PAaTPOPOTI ede. oss 2. oe: 0 1 0 0 0 1 2 al 0 0 
ETO POC ei sass cos sien Se ee 0 0 0 0 0 0 0 0 0 0 
Dactylopodite............. 0 0 0 0 0 0 0 0 0 0 
Oblates sescete ons 7 9 | 4 es 8 3 6 2 4 
TABLE 7.—DISTRIBUTION OF CYSTS IN E£. JAPONICUS FROM TOMIOKA, 


TOKUSHIMA PREFECTURE 


Total Cysts Cysts in Total Cysts Cysts in 
Number Number in Muscles, . Number Number in Muscles, 
| of Cysts Gills ete. of Cysts Gills ete. 
_ Se a ) 
1 7 7 ce 16 106 106 ane 
2 5 5 bite 17 ne ae. ae 
3 5 5 Mare 18 ‘s ae see 
4 98 } 81 64* 19 vias <a ame 
5 193 | 69 124 20 21 21 ee 
6 } 159 67 92 21 et 11 Sie 
7 40 8 82 22 36 36 Ey 
8 2 2 ie 23 48 48 a 
9 161 40 121 | 24 16 16 mee 
10 90 20 70 25 22 22 ae 
11 3 3 Be 26 ue ie Sai 
12 11 11 ate 27 35 85 Se 
13 7 24 52 28 12 12 sé 
14 51 33 18 29 53 53 siete 
15 17 17 HA 30 390 | 133 257 
* Three in liver. 
TABLE 8.—FROM THE ABOVE TABLE 
INUINDEEY OL CEEDS CXAMINCG 2%, 3/5 satvics s <coadsis oes dec caves soc: oe ese te 30 
Number:ot crabs infected with /Cysts...2: 5 .26s- cms +ccceouwbine nieces mace 26 
POTCOMLASO OT MIME CCG! CLADE: 2 so nels ccaicus seal ove ate somal petro spac 86.6% 
MIGUEL HIM DCUMOt: CYSCB. «2.0 '<a/e's cadence cemew dene ate Noe meee icles 1,668 
Average number of cysts in one Crab............cceccccccscecceccvcse 64.1 


TABLE 9.—DISTRIBUTION OF CYSTS IN THE MUSCLES AND THE HYPODERMIS 


OF ONE E. JAPONICUS HAVING 390 CYSTS IN ALL 
(NO. 30 OF TABLE 7) 

| First Leg Second Third | Fourth Fifth Total 

(Forceps) ) Leg Leg Leg Leg 
Body muscles attached to 49 28 28 9 12 126 
[achiopodite;. <<. cssees ss Ae 4 0 2 6 
Meropodite:. «2, :sseac sees 10 11 at 16 22 75 
Oarpopoulta.n... cvs sentence 10 8 9 6 0 28 
PLOPOGIGE: 2 taste hasteocens 9 2 6 3 2 22 
Dactylopodites. (osc .cenese 0 0 0 0 0 0 
TOA] .sscccspavaceaseis 78 48 59 36 36 | 257 


; 


YOSHIDA—INTERMEDIATE HOSTS OF LUNG DISTOME 115 


Morphology of the Encysted Larvae —The encysted larvae found 
in the above named crabs are almost spherical or rarely elliptical in 
shape, measuring from 0.25 to 0.55 mm. in diameter. The fully 
grown cysts vary between 0.30 and 0.55 mm. The wall of the cyst is 
a transparent chitinous membrane of tolerable thickness. In the fully 
developed larva in a cyst one may easily and distinctly recognize the 
organs, for example, both oral and ventral suckers, and alimentary 
tract including pharynx, esophagus, intestinal coeca, and excretory ves- 
icle. The oral and ventral suckers are of nearly equal size, the former 
is distinctly visible when the worm is moving and the latter becomes 
somewhat indistinct on account of being obstructed by an extension of 
the excretory vesicle. The pharynx is small, the esophagus short and 
the intestinal coeca run posteriad in a strongly winding course along 
both sides of body, ending blindly at or near the posterior end. The 
excretory vesicle occupies nearly all the space between the intestinal 
coeca. 

The parenchymous tissue of the body is tinged with light red pig- 
ment so that a cyst containing a larva is easily recognizable even in 
liver or in muscle. This pigmentation is very convenient in searching 
for the cyst. The young worm just out of a cyst is generally elongated 
oval in shape. Under a slight pressure one may more clearly observe 
all the internal organs, the minute spines with which the body surface 
is provided and the excretory vesicle contracted in discharging its 
contents. 

Animal Experiments with Encysted Larvae—tn the feeding exper- 
ments I have used young cats and dogs. These animals were all 
brought from uninfested districts and were carefully examined that 
none of them was infected with lung distomes. Some of them have 
already died or been killed and used for study of the development of 
the young worms in these hosts, but the others are yet at the present 
under experimentation. I will here describe the results of some of my 
experiments. 

(A) Young cat. The animal was fed with 20 cysts on July 26, 
80 on the 28th, and 130 on August 2. These cysts were all taken from 
the gills of E. japonicus collected from Tokushima prefecture. The 
cat died August 10, having passed 16, 14 and 9 days after the first, sec- 
ond and third feedings, respectively. Before her death she was 
extremely anemic and atrophied. In dissection the next day, numerous 
cestode larvae were found in the body wall as well as in the. body 
cavities, abdominal and pleural. Some portions of the body wall occu- 
pied by the worms had suppurated. It was easy to believe that the 
anemic and atrophic symptoms and consequently the death of the 
cat were caused by the presence of these larval cestodes. 


116 THE JOURNAL OF PARASITOLOGY 


In the abdominal cavity I found 18 young worms floating in serous 
fluid and adhering to the omentum, mesentery, and inner surface of 
the abdominal body wall, and in the pleural cavity 16 worms in serous 
fluid and on the pleural membrane. These young distomes in the body 
cavity were all nearly equal in size, measuring from 1 to 2 mm. in 
length in compressed specimens. The lungs were not yet occupied 
by the worms. 

(B) Young cat. Fed with 80 cysts August 7 and killed on 17th 
of the same month, 11 days after feeding. All the cysts were collected 
from the same locality given in the preceding example. I found 5 
young worms in the abdominal cavity and 6 in the pleural cavity. 
These worms measure about 1 mm. in length and 0.5 mm. in breadth. 
(Figure 6.) 

(C) Young dog. Fed with 33 cysts August 14, 46 on 17th, 90 on 
28th, 50 on 30th, and 32 on September 1. The cysts of the first and 
the second feedings were obtained from EF. japonicus of Tokushima 
prefecture and all the remaining cysts were taken from S. dehaani 
of Ebie, a suburb of Osaka. The dog died September 29, having 
passed 46, 43, 32, 30, and 29 days after each corresponding feeding. 
Some inflammations were observed here and there in the inner sur- 
face of the intestinal wall. This seemed probably to have been caused 
by the action of the young worms of the lung distomes. I found 
also a great number of Dipylidium caninum and a few Ankylostomuii 
canimum in the small intestine. Omentum and the mesentery were 
slightly congested here and there. There were numbers of perfora- 
tions on the inner surface of the abdominal body wall. Around the 
perforations more or less hemorrhage was observed between the mus- 
cular layers of the abdominal wall. These perforations were evidently 
produced by the young worms which were found everywhere in the 
serous fluid or on the surface of various organs in the abdominal cay- 
ity where I had obtained 30 young worms. A number of perfora- 
tions was observable on the diaphragm through which the young 
worms seem to have passed from the abdominal to the pleural cavity. 

In the pleural cavity 43 young worms were obtained in the serous 
fluid and on the surface of the various organs: lungs, heart, and 
pleural membrane. The size and the shape of the worms in the body 
cavities were extremely variable according to the state of contraction. 
In fixed specimens, the length varies from 1.5 to 5 mm. and the breadth 
from 0.5 to2 mm. I found a good specimen which is most favorable 
to demonstrate that the worm in the pleural cavity enters the lungs 
from its surface by perforating. Numerous worms in various stages 
of development were observed in the lungs. The worms of large size 


YOSHIDA—INTERMEDIATE HOSTS OF LUNG DISTOME 117 


in the lungs were plainly observable from the outside or easily felt by 
touching on the surface of lungs. 

(D) Young cat. Fed with 4 cysts June 15, 1 on 16, and 3 on 17, 
and was killed on August 17. It was 62 to 64 days after the feed- 
ings. All the cysts were taken from the liver of S. dehaani. In the 
dissection of this animal I found 2 young distomes, one in the pleural 
cavity, the other in the right lung. The dimensions of this worm 
were about 4 mm. long and 2 mm. broad. The worm in the lung 
was not yet matured. 

From my observations in the above experiments, I have learned 
the general course which the young worms travel from the intestine 
to the lungs of the host. Briefly stated, it is as follows: 

The encysted larvae swallowed by a host come out of their cysts 
in the stomach or intestine through the action of gastric or intestinal 
juice. The larvae coming out of the cysts are actively mobile. They 
pierce the wall of the intestine. They stay for some time in the abdom- 
inal cavity and wander about here and there; thence they pierce 
through the diaphragm to enter the pleural cavity. Here again they 
stay for some time and finally penetrate into the lungs from their 
surface. 

In addition to this general course, the young distomes in the abdomi- 
nal cavity may also penetrate the abdominal wall and move around in 
the muscular layers of the connective tissues, as stated above in 
Case C. Some worms in the pleural cavity may proceed cephalad, 
taking their course through the loose connective tissues along the 
esophagus or the blood vessels. Thus the young worm of the lung 
distome has evidently a wandering power by which it can pass through 
the muscular and connective tissues. It is obvious that the discovery 
of the wandering character of the young worm throws light on the 
accounts of cerebral and spinal paragonimiasis which are attributed 
to the lung distomes. 

October, 1915, 


118 THE JOURNAL OF PARASITOLOGY 


EXPLANATION OF PLATE 


Fig. 1—Cyst attached to lobule of liver of S. dehaani. x60. 


Fig. 2—Longitudinal section of the right third leg of FE. japonicus, show- 
ing distribution of cysts. Natural size; b, basipodite; c, carpopodite; d, dactylo- 
podite; g, gill; h, hypodermis; i, ischiopodite; k, muscle; 1, encysted larvae; 
Pp, propodite. 


Fig. 3—Cyst from the gill of E. japonicus. x60; i, intestinal coeca; e, 
excretory vesicle; 0, oral sucker; v, ventral sucker. 


Fig. 4—Larva escaping from cyst. x60. 
Fig. 5.—Larva coming out of cyst; slightly compressed specimen. x60. 


Fig. 6—Young worm in abdominal cavity of young cat. x60; c, excretory 
canal; note also intestinal coeca. 


PLATE 


<a 


Fig. 5 


Fig. 4 


GONGYLONEMA IN THE ROLE OF A HUMAN 
PARASLIE * 


Henry B. Warp 


Through the kindness of Dean Charles E. Brookover of the Uni- 
versity of Arkansas Medical Department, Little Rock, I received 
recently a specimen of a nematode which has not heretofore been 
recorded as a human parasite. The worm was removed from the 
lip of a girl. The conditions surrounding the case are reported as 
follows by the attending physician, Dr. Robert Lee Covington of Jeffer- 
son, Ark., to whom I am indebted not only for the case history cited 
here, but also for replies to several inquiries on the matter necessary 
to perfect the record for publication. Dr. Covington’s report is as 
follows: 


Miss —, 16 years old. I was called to see her September 3. Found her 
suffering with considerable digestive disturbance, vomiting continuously for two 
or three hours; temperature 101.5 F. She felt chilly. I administered calomel 
followed with castor oil. Later I gave her full doses of turpentine and castor 
oil. The fever woukd come and go and for this I gave quinin. She improved 
some; later the fever abated entirely, but some digestive disturbance still per- 
sisted. She was very anemic and irritable. September 12 I dismissed her. 

I was again called October 1. She wanted me to see what that was running 
around under the skin of her lip, as she expressed it. I thought it was pure 
imagination. She said she could see it by looking in the mirror; she said it 
looked like a worm, and at times she could feel it leave her lips and go back 
as far as the fauces. I examined the lower lip where she said it had stopped. 
I discovered the outline of what looked like a worm about three quarters of 
an inch in length and about the size of a No. 60 sewing thread; it was just 
beneath the mucous membrane. I inserted a needle under it and pulled out 
a little of one end, but before I could grasp it, it got away, moving back near 
the corner of the mouth. There I ran the needle under it midway between the 
ends and pulled upward, bringing a small loop through the mucous membrane. 
It held on so tight that the ends on each side of the needle cut their way out. 

Up to this time she was extremely anemic and very cross and irritable, but 
after the removal of the worm she rapidly improved, her disposition changed, 
and now she is not like the same girl. She seems to be sound and well at 
this date. 

Dr. Rogert LEE CoviNcToN, 

Jan. 26, 1916. Jefferson, Ark. 


In response to specific questions Dr. Covington stated that the 
worm moved up and down in the tissues three or four times, extending 
its migrations from the lips at least as far back as the fauces. It was 
actually seen by the patient three times and she reported it positively 


* Contributions from the Zoological Laboratory of the University of Illi- 
nois, No. 59. 


120 THE JOURNAL OF PARASITOLOGY 


to her father the day before its removal. It could be discerned clearly 
enough to tell that it was a worm; in color it was a little lighter than 
the mucous membrane. During its movements the worm was definitely 
sensible to the patient only when it approached the skin and it was not 
seen or felt in this organ anywhere except on the inside of the lips. 
It. was seen only inside the mouth through the thin mucous membrane 
of that cavity and so far as known did not approach the external skin 
save at the inner border of the lower lip. 

From this record one may safely conclude that it migrated to and 
fro through the loose connective tissue beneath the oral mucosa and 
endeavored to move up into the firmer derma only when it approached 
the lips. From the fact that it evaded the surgeon for a time, one can 
see that the movement was free and active. 

Subsequently Dr. Covington wrote regarding the patient: “She has 
lived here all her life, has not traveled in any other country. ial 
Sanitary conditions are not just what they ought to be. They get 
drinking water from a well that is very shallow and fills up to the top 
when it rains heavily.” 

On preliminary examination it appeared that the specimen had 
suffered somewhat in the handling incidental to its forcible removal, 
and was not in such histological condition that much could be said 
definitely regarding its internal structure. 

Under a dissecting lens the specimen (Fig. 1) is seen to be a nema- 
tode worm, light brown in color, semitranslucent, and loosely coiled, 
though both ends are nearly straight. The worm measures 42.1 mm. 
in total length and is of nearly equal diameter throughout, tapering 
only a little near the two ends which terminate rather abruptly. 

The anterior end appears to taper more toward the bluntly round 
tip and for a space of 1.4 mm. from the end the surface is orna- 
mented by various cuticular outgrowths like scales or tubercles. These 
are arranged in somewhat definite fashion (Fig. 2). A cuticular ridge 
extends along the lateral line of the body starting at a point about 
0.25 mm. from the extreme anterior tip and running back about 1.5 
mm. In the anterior region this ridge is slightly irregular but nearly 
equal in height at all points ; further back it is divided by deep indenta- 
tions into long ovals. At the place where it starts the diameter of the 
body is about 0.1 mm. and at its posterior end the body has increased 
in diameter to 0.19 mm. 

In the submedian lines are rows of scales, plates, or tubercles that 
start just behind the front end of the ridge just described and extend 
backwards only about 1.1 mm. in definite and regular order. These 
tubercles stand in a linear series so close together that they are prac- 
tically continuous even though separated from each other by a dis- 
tinct boundary line. They vary in form and size but on the average 


WARD—GONGYLONEMA A HUMAN PARASITE 121 


are nearly square in profile and measure 0.022 mm. in height and 0.025 
to 0.3 or rarely 0.045 mnt. in length. In front of this close-set series 
and of the cuticular ridge one finds a number of isolated cuticular 
bosses which in part line up with the ridge or the series and in part 
do not. Between the ridge and the series are a few isolated scales. 
These are irregular, detached, and on the average larger than the 
tubercles in the regular series. 

The anterior tip of the body possesses a small infundibuliform 
crown surrounding the mouth which may represent a group of lips 
with some papillae. From the orifice a capilliform esophagus extends 
posteriad and disappears from view behind the anterior tip of the 
cuticular ridge. In the mid-ventral line about 0.6 mm. from the 
anterior tip is a peculiar papilla that probably surrounds the excretory 
pore. The location of the end of the esophagus is difficult to deter- 
mine precisely ; it appears to be about 7.5 mm. from the anterior end. 
Here the diameter of the body is 0.23 mm. ; a measure which is main- 
tained with only slight variations from 0.22 to 0.24 mm. approxi- 
mately, throughout the entire length until the caudal region is attained. 

The vulva is a ventral, slightly prominent pore with raised lips, 
located 2.15 mm. from the posterior tip of the body. Even behind it 
the body retains its uniform caliber of approximately 0.23 mm. until 
close before the anus or about 0.25 mm. from the exterior tip. This 
region is a little damaged by handling, but shows distinctly a slight 
concavity on the dorsal surface making the bluntly rounded posterior 
tip of the body turn a bit upwards. At the anal orifice the body 
measures 0.09 mm. in diameter, which is evidently somewhat less 
than normal owing to the injury already noted. The taper begins 
only a short distance anteriad to the anus, and if the specimen had 
been uninjured, would have been regular apparently from that point 
to the rounded tip. There is certainly no abrupt change in diameter 
either before or behind the anus. 

There can be no doubt that this specimen belongs to the genus 
Gongylonema in the family of the Filariidae. Gongylonema is a 
nematode peculiar in habit in that it forms a sinuous gallery in the 
mucous epithelium of the esophagus. The worm lies in this tunnel 
with the anterior region alone projecting from it. With a single 
exception all species are found in mammals. The body is very long 
and thread-like, showing a slight taper toward both extremities. The 
characteristic feature of this genus is the presence on the anterior 
region of a considerable number of scales or tubercles which are 
elevated thickenings of the external cuticula only. These differ in 
size, number, and arrangement in different species. Cuticular folds 
stand out from the surface along the median or lateral lines in the 


122 THE JOURNAL OF PARASITOLOGY 


anterior region, and form conspicuous features in the external aspect 
of the worm. These also vary in number, form, and extent in various 
species. The vulva is located near the posterior end, and its precise 
position as well as the form and size of the tip of the tail are valu- 
able in determining the species of female specimens. 

Two species of Gongylonema are common to domestic animals in 
the United States, and fall under suspicion as possible occasional 
parasites of man. These are G. scutatum and G. pulchrum. Of G. 
ingluvicola Ransom from chickens it need only be said that the struc-- 
ture of the adult is too dissimilar to allow of the surmise that this 
specimen is an erratic individual of that species. But the other two 
are much alike in structure and are abundant in some parts of this 
country, if not in most regions; they are also both very similar to the 
specimen under consideration. 

G. scutatum was recently described with some care by Ransom 
(1911) who declares that it is very common in this country and can be 
found in a large percentage of cattle and sheep slaughtered at 
abattoirs. 


G. pulchrum is known as a parasite of the hog in Europe, and has 
been found also in North Africa by Seurat (1912). It does not 
seem to have been reported in print from the United States previous 
to this year when it was briefly mentioned by Ransom and Hall (1916) 
in connection with the discussion of experiments on the life history of 
G. scutatum. They have found it to be frequent in the vicinity of 
Washington, D. C., and it doubtless occurs abundantly in the pig in 
other regions as well. Dr. B. H. Ransom was good enough to send me 
specimens for comparison. These came from the collections of the 
Bureau of Animal Industry and were taken from a hog at Bethesda, 
Md., in January, 1914. 

These two species are very similar, and in the present case it is 
impossible to assert positively which one is the specimen under con- 
sideration. It agrees in length with G. pulchruwm and as nearly as can 
be ascertained also in the character of the female genital organs of 
which a comparative study has been published by Seurat (1912). The 
caudal tip is also slightly concave on the dorsum as has been described 
in that species. However, this specimen is a trifle over ordinary length 
for G. pulchrum, although it has not yet reached full sexual maturity. 
But the genital pore is salient, although not notably so, a feature 
which is said to be characteristic of G. pulchrum though wanting in G. 
scutatum. On the whole, I am inclined to determine the specimen 
from man as G. pulchrum. The common ascarid of man, the stomach 
worm, A. lumbricoides, is also a parasite of the pig in which it occurs 
abundantly in all parts of the country. 


WARD—GONGYLONEMA A HUMAN PARASITE 123 


The specimen under consideration manifested a habit that so far 
as known is not characteristic of the genus to which it must be 
assigned. It was engaged in active migrations through the subdermal 
connective tissue, and the patient was conscious of the fact. It had 
approached near enough to the surface to be seen and correctly diag- 
nosed in form, and had wandered away into deeper tissues. The 
worm may have been thus active over a period of a month or less. It 
moved through the connective tissue rapidly enough to render its cap- 
ture and removal a matter of skill and dexterity. Such performances 
are not reported in other accounts of the genus, which as already noted 
is found in the mucosa of the esophagus in its normal host, and has 
not been collected from other organs or regions. The presence of 
the chicken species in the wall of the crop is not a real departure 
from this general habit for it occurs there in the mucous lining 
according to Ransom’s report. It is, of course, possible that these 
forms regularly inhabit the subdermal connective tissue during a period 
of their life cycle and appear in the esophageal mucosa at the time 
of sexual maturity; or such an occurrence may be exceptional but 
actual in the normal host also. In that event it would readily escape 
notice under usual conditions. There is no basis for deciding whether 
such wanderings are usual but unnoted heretofore in the normal host, 
so rare in it as to have evaded observation thus far, or peculiar to the 
human host or to this particular case. 


The habit recalls very strikingly the wandering through subdermal 
tissues of the African eyeworm, Filaria loa, which has received its name 
from its frequent appearance in the subcutaneous connective tissues 
near the eyeball. It is also well known to occur in the same tissues 
elsewhere in the body. Among the cases on record are a few which 
report its removal from the lip or near the same, but it occurs habitu- 
ally near the external skin and is not reported from the vicinity of 
the mucosa. Yet it is not at all impossible that in some instance where 
no careful examination was made a specimen of Gongylonema has 
been interpreted as the loa. The length of the specimen reported 
here and its appearance agree in general with the loa, and while it 
is not quite so heavy nor so opaque, the two might readily be con- 
fused if no careful microscopical examination were made; all the 
more so since the loa is the only nematode which has been known to 
move about in subdermal tissues and that striking habit would have 
been taken to indicate the type of parasite at hand. 

There are other records of parasitic nematodes from man in this 
country that should be brought briefly under consideration in connec- 
tion with the case in hand. In 1850 Dr. Joseph Leidy of Philadelphia 


124 THE JOURNAL OF PARASITOLOGY 


described a human parasite as Filaria hominis oris from a specimen 
in the collection of the Philadelphia Academy labeled “obtained from 
the mouth of a child.” 

This account of Leidy has always been difficult to interpret. His 
conjecture that it was a young Dracunculus or Medina worm has been 
generally rejected and other suggestions are not entirely satisfactory. 
The location suggests that the specimen may have been a Gongylonema, 
not the species here reported for that is much too short, but G. 
scutatum of cattle and sheep which is of the right dimensions. Leidy 
was a very accurate observer and it is improbable that he would have 
overlooked the cuticular tubercles on the anterior end if such had 
been present. The condition of the specimen might have been such 
as to prevent the diagnosis of these structures, but on the whole the 
case can hardly be assigned to Gongylonema even tentatively. 

The source of the infection in the case at hand can not be positively 
ascertained and yet recent studies suggest a very probable explanation. 
Thanks to the careful work of Ransom and Hall which has extended 
over several years it is now known that the larvae of G. scutatum 
occur in various species of dung beetles and that they have been raised 
experimentally not only in these but also in croton bugs. Furthermore, 
eggs of a Gongylonema, most probably G. pulchrum from the eso- 
phagus of a hog, were fed to croton bugs and at the close of a 
month embryos were found encysted in the final stage and ready for 
transmission. 

In the present case it is easy to see how an infected insect, very 
likely a croton bug, might have been ingested whole, or some frag- 
ments of it included by accident in meal, flour, milk, or other materials 
used in cooking. Such sanitary mishaps are very common with poorly 
prepared food, and thus the infection of the human host would have 
been achieved. The sanitary conditions noted in the clinical history 
of the case favor such an occurrence. 

The duration of the infection can be estimated as at least a month 
if the clinical symptoms stand in any connection with the parasite. 
No data have been found to show the rate of development of these 
worms in the final host but if one can infer safely from other nema- 
todes it is likely that this specimen was in the human host more than 
a month and that the symptoms did not manifest themselves until it 
had attained a certain measure of growth. These symptoms were so 
definite and terminated so promptly with the removal of the parasite 
that etiologic significance can hardly be denied to the worm. 


SUMMARY 


A specimen of Gongylonema, probably G. pulchrum, has been 
recovered from man. This species is normally a parasite of the pig. 


PLATE 


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a 
RS 


H 


\ 
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\ 
4 
1 


at! 


wren 
me 


(TED CSA CD 


St are 


te 


(Tete catty bel yet b 


EXPLANATION OF FIGURES : 
Fig. 1—Gongylonema (pulchrum?) from human host. Camera drawing. X 15 


Fig. 2—Antericr end of same specimen. Camera drawing. X 140. 


Ae 
ert Oi 


"iy ‘Ls as. el 


WARD—GONGYLONEMA A HUMAN PARASITE 125 


Infection of the human host was brought about probably by the 
ingestion of larvae in the imfective stage which had developed in some 
insect. Very likely the croton bug, known by experiment to be able 
to serve as intermediate host for this species, was the source of the 
infection which might readily occur by accident. 

The presence of the parasite was accompanied by clinical symp- 
toms indicating marked digestive and nervous disturbances, associated 
with anemia. These symptoms disappeared with the removal of the 
worm. 

The parasite displayed a tendency to wander through the sub- 
mucosal connective tissue from the lips to the throat. 


Papers CITED 


Leidy, Jos. 1850. Description of Three Filariae. Proc. Acad. Nat. Sci., 
Phila., 5: 117-8. 

Neumann, L. G. 1894. Sur le genre Gongylonema Molin. Mém. Soc. Zool. 
France, 7: 463-473; 4 figures. 

Ransom, B. H. 1904. A New Nematode (Gongylonema ingluvicola) Para- 
sitic in the Crop of Chickens. Bur. An. Ind., Circ. 64. 1911. The Nematodes 
Parasitic in the Alimentary Tract of Cattle, Sheep, and Other Ruminants. 
Bur. An. Ind., Bull. 127. 

Ransom, B. H., and Hall, M. C. 1916. The Life History of Gongylonema 
scutatum. J. Parasitol., 2: 80-86. 

Seurat, L. G. 1912. Sur l’appareil génital femelle des Gongylonemes. C. R. 
Soc. Biol., 73: 276-9; 5 figures. 


ARE SARCOSPORIDIA ABERRANT FORMS OF 
CNIDOSPORIDIA OF INVERTEBRATES? 


B. GALLI-V ALERIO 


University of Lausanne 


In the year 1910 v. Ratz stated that Sarcosporidia are very near to 
the Nosematidae. In an interesting paper Darling (1915) writes: 
“On account of the facility with which herbivora may obtain and ingest 
invertebrates infected with Neosporidia, but more particularly flowers, 
leaves, etc., or water contaminated with droppings of bees, moth lar- 
vae, fly droppings, etc., or other material containing Neosporidia 
spores, is it not possible that Sarcosporidia may be side-tracked vari- 
eties of some of the Neosporidia of invertebrates which have invaded 
the musculature of a hospitable though by no means definitive host 
and, being unable to continue further their life cycle, have escaped 
from a compromising and aberrant position?” 

This opinion of Darling was supported by the experiments of 
Scott (1915) who concludes his paper with the following words: “All 
of the evidence favors the view that the sheep is not the definite host 
of S. tenella and, therefore, is in accord with Darling’s suggestion that 
the muscle parasites of vertebrates are aberrant forms.” 

In such an order of ideas it is perhaps interesting to note observa- 
tions which seem to me to support the hypothesis that Sarcosporidia 
are identical with Cnidosporidia. 

In 1896 Piana, in a paper on the life cycle of Balbiania gigantea-S. 
tenella, states that in the cultures of this parasite on sterilized moist 
filter paper, moist earth, gelatine prepared with Focus crispus at a tem- 
perature of 18 to 25 C. in twenty-five to sixty days the spores give 
bodies which develop more and more and present a nucleus and ame- 
boid movements. The movements cease after some days and the 
amoeba assumes the form of a cyst. Piana tried to infect a sheep 
per os with this material, but with negative results. I may add the 
macroscopic examination made with a view to finding cysts of Bal- 
biania, was confined to the esophagus. 

After seventeen years I repeated (1913) the experiments of Piana 
with S. muris, making cultures of this on sterilized filter paper 
moistened with physiological salt solution at a temperature of 20 C. 
After eight days I found in this culture amoeboid bodies showing slow 
movement at a temperature of 20 and 37 C. This amoeba had a fine 
granulated protoplasm, a nucleus, and clear vacuoles. Colored with 
Leishman or Giemsa the protoplasm assumed an azure color, and the 


GALLI-V ALERIO—ARE SARCOSPORIDIA ABERRANT FORMS 127 


nucleus a red color. After thirty-two days the cultures presented only 
encysted forms. With this culture I inoculated a white rat and a 
black mouse in the muscles of the thigh but without result. The inocu- 
lation of a white rat with S. muris was negative, as was also the infec- 
tion per os of a guinea-pig with the same Sarcocystis. Neither Piana 
nor I affirm that the amoebae were positively derived from the spores 
of Sarcocystis, but we stated that it was very probable; and the obser- 
vation of Erdmann (1914) that in the intestinal cells of a mouse 
infected per os with Sarcosporidia appear little amoeboid bodies, seems 
to support our experiments and the fact that Sarcosporidia are iden- 
tical with Cnidosporidia. This idea was also expressed by Auerbach 
(1910) who says: “Mit der Tatsache des vorhandenseins einer Polkap- 
sel bei Sarcosporidiensporen fallt ein wesentlicher Unterschied gegen- 
uber den Cnidosporidian fort und es liegt kein zwingender Grund vor, 
sie noch von diesen zu trennen.” 

If it is possible to demonstrate that spores of Sarcosporidia produce 
amoebae, the identity of the two orders is more probable. The draw- 
ings of the amoebae of Myxidium inflatum, leptotheca macrospora 
and Myxidium bergense in Auerbach (1910: 10, 79) are very likely 
the drawings of amoebae of S. muris shown in my paper. 

Given the spores of the Sarcosporidial amoebae, the infection with 
Sarcosporidia must be analogous with the infection with Cnidosporidia. 
The experiments of Thélohan and particularly those of Auerbach on 
the development of the Cnidosporidia in the intestine of fishes prove 
that the spores of this parasite produces amoeboid bodies in the duo- 
denum. It is probable that the spores of Sarcosporidia must also pro- 
duce amoebae in the intestines and perhaps in Piana’s experiments 
the amoebae were destroyed in the stomach. In fact Auerbach failed 
to observe a transformation of spores of Cnidosporidia in the stomach 
of fishes. I think it would prove an interesting experiment to make 
cultures from the great cysts of S. tenella, and to introduce the amoebae 
directly into the intestines of sheep. With such a method of infection 
it is perhaps possible to secure much more positive results than to 
work with spores of Sarcosporidia which as Auerbach states are in 
some cases probably not ripe. 


SUMMARY 


1. The observations of Piana and Galli-Valerio to the effect that 
spores of Sarcosporidia produce amoebic bodies in cultures, more 
closely relate the Sarcosporidia to the Cnidosporidia. 

2. If true that Sarcosporidia are only aberrant forms of Neo- 
sporidia of invertebrates, then the hypothesis of Darling becomes 
more probable. 


128 THE JOURNAL OF PARASITOLOGY 


PAPERS CITED 


Auerbach, M. 1910. Die Cnidosporidien. Leipzig, 261 p., 4 pl. 

Darling, S. T. 1915. Sarcosporidia Encountered in Panama. J. Parasit., 
1 :113-120. 

Erdmann, Rh. 1914. The Schizogony of Life Cycle of Sarcocystis muris. 
Proc. Soc. Exper. Biol. Med., 11 :152-153. 

Galli-Valerio, B. 1913. Notes de parasitologie et de technique parasitologique 
et observations sur quelques tumeurs des animaux. Cent. Bakt. i, Orig., 69 :496-504. 

Piana, G. P. 1896. Fasi evolutive dei sarcosporidi. Mod. zooiatro, Torino, 
6 :145-147. 

Ratz, von S. 1910. Ueber die Struktur dei Sarkosporidienschlatiche. Arch, 
weiss. prakt. Tierheilk. Suppl. 36 :573-589. 

Scott, J. W. 1915. Some Notes and Experiments on Sarcocystis tenella, 
Railliet. Jour of Parasit., 2 :20-24. 


THREE NEW GREGARINES FROM MARINE CRUSTACEA * 


MInNIE E. Watson 
Oyster Bay, Long Island, N. Y. 


The protozoan parasites described here were studied during the 
summers of 1913 and 1914 at the Biological Laboratory of the Brook- 
lyn Institute, Cold Spring Harbor, L. I., and I wish to thank Dr. C. B. 
Davenport, the director, for kindly affording me the privileges of the 
laboratory. 

All the hosts were found on the Sand Spit, a narrow peninsula 
half a mile long separating the outer and inner harbors. The area 
is thus geographically very restricted, and although the hosts are simi- 
lar and the possibility of all of them being infected with the same 
parasite considered, yet there is abundant evidence to warrant regard- 
ing the three forms described below as separate species. 


Frenzelina delphinia nov. spec. 
Figures 1 to 8 


The host of this gregarine is the large white sand flea, Talorchestia 
longicornis, which is found in fine sand between tide marks. Fleas 
were taken from the eastern end of the Sand Spit, and also from loca- 
tions along the road to Lloyd’s Neck, from Huntington Beach, and 
from Northport Harbor; and hosts from all these localities were found 
to be infected. 

The parasites were present in 30 per cent. of the 260 intestines 
examined, the number in an infection varying from one to ten in 75 
per cent. of the hosts and from ten to 500 in the other 25 per cent. 

The sporonts (Figs. 1 and 21), which live free in the lumen of the 
upper part of the intestines, are small, the average of fifty large indi- 
viduals being 110 in length and 60p in width. The largest individual 
seen measured 115y in length and 64» in width. The ratio of length 
of protomerite to total length of sporont is about 1:4; the ratio of 
width of protomerite to width of deutomerite about 1:1.5. A table of 
a few measurements is given at the end of this section. 

The sporonts are biassociative when mature and the two members 
of an association do not differ materially in size; either the primite 
or the satellite may be a little longer or broader than the other. The 
sporonts are stout bodied, being less than twice as long as wide.. The 


* Contributions from the Zoological Laboratory of the University of Illinois, 
under the direction of Henry B. Ward, No. 60. Also from the Biological 
Laboratory, Cold Spring Harbor. 

*For explanation of special terms used see description of Figure 1. 


130 THE JOURNAL OF PARASITOLOGY 


protomerite is cylindrical and broadly rounded in front and is about 
three fourths as high as wide. There is often a slight constriction at 
the septum which separates the protomerite and deutomerite. The 
deutomerite is doliform, two thirds as wide as long, widest through 
the middle portion, and terminates in a broadly rounded extremity. 
There is a small conspicuous papilla at the anterior end of the pro- 
tomerite of the young sporonts which persists, although reduced in 
size, in the adults. In the satellite of an association it becomes a struc- 
ture for the attachment of the two sporonts by projecting upward and 
forming a small indentation in the posterior end of the primite (Fig. 
4). Hydrochloric acid, either dilute or concentrated, fails to dislodge 
the two sporonts, but sodium hydroxid or ammonium hydroxid, in 
solution, readily disassociates them. 

The endocyte of the mature sporonts is light brown in color and 
in full-grown animals fairly dense, and the nucleus not visible. Young 
sporonts are less dense and the color paler. The satellite of an asso- 
ciation is generally less dense than the primite, and for this reason its 
nucleus is often visible in vivo when that of the dense primite is not. 
The protomerite of a sporont is slightly less dense than the deutomerite 
and contains fewer protoplasmic granules; the granules are slightly 
larger than those in the deutomerite and the endocyte paler in color. 
When the host has just been taken and has probably fed the night 
before, full grown sporonts are dense; when, however, the fleas have 
been kept in damp sand for a few days without food, the parasites are 
likewise deprived of nourishment and become pale in color and the 
number of protoplasmic granules present, hence the density, is greatly 
reduced. The addition of a drop of iodin solution renders these pale 
parasites visible but of course kills them. 

The nucleus of the sporonts is generally visible in vivo only as a 
lighter indefinite area although in young and starved sporonts it is 
often visible. It is large and spherical and contains one or two large 
homogeneous karyosomes. The epicyte is thin and fragile and is 
marked with longitudinal striations only visible under an oil immersion 
lens. 

Cross sections made of the intestine of the host reveal the fact that 
development is intercellular as in the family of the Stenophoridae. 
This is the first genus of the family Gregarinidae which has been found 
to develop in this manner, the other members possessing an epimerite 
which alone is attached to the host cell, the remainder of the tropho- 
zoite projecting into the lumen of the intestine. In the species here 
described, the whole trophozoite is embedded in the epithelium (Figs. 
3 and 6). When stained with Delafield’s hematoxylin, the cells of the 
epithelium become purplish-blue, while the parasites stain less deeply 
and the color is a clear homogeneous blue with a darker blue nucleus. 


WATSON—GREGARINES FROM MARINE CRUSTACEA 131 


The animals are capable of movements both of bending and of 
gliding progression. The sporonts are able to move through a narrow 
place in the manner employed by an amoeba. When the host intes- 
tine is first opened, a mosaic of inert distorted individuals or associa- 
tions lying near the epithelium is often revealed ; when, however, water 
or normal salt solution is added, the smooth, regular contour is quickly 
restored. Normal salt solution stimulates movement and sporonts 
often remain alive and motile for an hour and a half. Sea water has 
the same effect. As motion tends to be retarded at the end of this 
time, weak tannic acid solution was added in a few instances and 
caused considerable acceleration of movement; plasmolysis, however, 
occurred inside of another half hour and the animals died. 

That transparent threads of mucus are present at the posterior end 
of the body was frequently attested by the fact that the animals were 
able to carry with them in their movements large or small masses of 
débris at a distance behind the body often as great as the length of the 
animal itself. A mass twenty-five times the volume of the gregarine 
itself was in one instance observed being carried along. 

Cyst formation was observed in material from the host intestine 
from its incipiency until rotation ceased. The adult sporonts free in 
the intestine which are ready for cyst formation become thickened and 
shortened, motion becomes sluggish, movement of progression ceases 
and that of bending becomes more active (Fig. 5). Concomitant with 
the revolving motion there occurs a deposition of gelatinous threads 
exuded from the body in fine concentric layers around the revolving 
mass. The sporonts become a spherical mass and the threads form a 
thick cyst wall. The rotating mass passes from the mid intestine to 
the rectum and ceases motion (Fig. 7). It begins here its development 
by the loss of the wall separating the two sporonts and the disintegra- 
tion of the two sporont nuclei. The protoplasm of the cyst collects in 
masses and on the periphery of each protoplasmic mass are formed 
spherical protuberances, the gametids. The cyst is now expelled with 
the feces. Cysts when expelled are somewhat opaque, tan in color, 
and average 80p in diameter, including the cyst wall. 

It is difficult to effect cyst development to completion by artificial 
methods. Marine bacteria seem to be virulent and to cause putrefaction 
or otherwise stop development in the early stages. Cysts were kept in 
the damp chamber in normal salt solution and a few yielded gametes 
upon being crushed when 24 hours old. The gametes were stained 
with safranin; they were large and nearly spherical, and no difference 
in size was observed in those from the same cyst. Some of the gametes, 
however, showed large deeply staining nuclei and others smaller nuclei 
which stained less deeply, probably because of their reduced chromatin 


132 THE JOURNAL OF PARASITOLOGY 


content. A difference in the staining reaction was also noted in the 
two sporonts in a cyst of about eighteen hours, the one sporont mass 
staining more deeply than the other. 

A very few cysts developed to completion and dehisced by simple 
rupture in 35 hours, but no well formed spores were present. 

Orchestia agilis, the smaller very common sand flea, was found in 
one instance to be infected with three nonassociative sporonts which 
agree in size and shape with the species described above. This flea 
was also heavily parasitized with an infusorian. Orchestia grillus from 
the roots of the eel grass (Juncus palustris) was not found to be 
infected with gregarines. 

The genus Frenzelina (Léger and Duboscq, 1909) has not hitherto 
been reported from the United States and only seven species have 
been described. The species described above is placed in this genus 
because (1) the sporonts are biassociative, (2) the cysts dehisce by 
simple rupture, (3) development is intercellular, (4) the apex of the 
protomerite is slightly papillated, (5) the parasite inhabits the intestine 
of a crustacean. 4 

I wish to designate the species, of which no previous record is 
found, Frenzelina delphinia. 

The fact that development is intercellular was not determined by 
Léger and Duboscq and this important fact should be added to the 
features which characterize the genus Frenzelina. 

A table of typical measurements, in microns, follows: 


Motamleneth association... <ccoss sn oe teens 215 215 210 


Primite : 

Moralsleneth sporont......... .«aseeeee eee ore 115 110 112 
Wenotheprotomerite ...... wears eee 20 24 27 
Were hisGeaLOmernite. . .... acs nce 95 86 93 
Wath pLOLOmenite. «. ...... 2.0 ciensen eee eee 39 35 43 
WitGEMCeHtOMmerite .« 2. <> conseeneemeieee 62 66 63 
Ratio length protomerite: length deuto- 

GACTILE MEA. 6 cia. b. so ooo 5 sista seh eee 1:4.7 1 :4.6 1:35 
Ratio width protomerite: width deuto- 

RACEALEMIPE I eis eos. «io sere ern 1216 1:18 1:15 

Satellite : 

Motalmlensthysporont. ..../.\...seseeeeeeee 100 105 98 
WenothiepsOtOMerite ..<<\<\: «6 tc.ecteseeels 20 20 20 
PSR MCHEOMICTIEG .. ..as's - dbs ease eee 80 85 78 
WHOLTMOEOLOMICLILG 5... ..-.ce. sa ees eee 37 40 40 
Wil thdelntomorite cs... ...eensnemeeeeee 55 56 60 
Ratio length protomerite: length deuto- 

GORD ./5 os OREO CEe eric -Soace Ibe 1:1.4 Lists 
Ratio width protomerite: width deuto- 

TTT EINE MEP MMO N ees e.o7s, 5 vd .0i6:0 Gaeta bratele ot oeetorere ihe he 1:1.4 115 
CSE eR EIARIELED. 2% 5 si000!4 ood ame oe 77 90 86 

IifitletstiATHTe Leis cc = arvi5 6a -on cee eerie 63 74 66 


Thickness transparent layer around cyst.. 7 8 10 


WATSON—GREGARINES FROM MARINE CRUSTACEA © 133 


Frenzelina olivia nov. spec. 
Figures 8 to 10 

The host of this species is the small littoral spider crab, Libinia 
dubia, which is abundant along the shores of Long Island Sound and 
its inlets. The parasite is found in the upper part of the intestine. It 
generally occurs in moderate numbers (10 to 100) but rarely an infec- 
tion of 1,000 or more is encountered. 

The sporonts are biassociative and average 80 in length and 35h 
in width. They are ellipsoidal in shape, rounded in front and rather 
blunt posteriorly. The protomerite is hemispherical, very slightly con- 
stricted at the septum. It is about one fifth the total length of the 
sporont and slightly papillate in the adults (Fig. 8); the younger 
solitary sporonts possess a conspicuous papilla (Fig. 10). The deuto- 
merite is but little wider than the protomerite (1:1.2) ; in solitary indi- 
viduals it is more broadly rounded at the posterior end than in those 
which are attached. The endocyte of the matttre sporonts is dark 
brown and very dense in the deutomerite; the protomerite is less dense 
and tan in color. At the anterior end of the protomerite is an orange 
colored disc. The nucleus is visible only in immature specimens. It 
is spherical and generally contains one large karyosome. 

Movement of progression is rapid and continues only for intervals 
of about two seconds. 

Cysts are spherical, dark brown in color, and from 45 to 60 in 
diameter including the enveloping wall. They occur in the posterior 
third of the intestine. No sections were made of the host intestine. 

This species is placed in the genus Frenzelina because (1) it is 
biassociative, (2) there is a papillated conspicuously differentiated 
apical area in the protomerite, (3) it is very similar to Frenzelina 
delphinia in form and location and both occur in hosts from the same 
habitat; (4) it is parasitic in the intestine of a marine crustacean. 

The larger spider crab, Libinia emarginata, which is found in 
deeper water and seldom comes near the shore, has been examined 
repeatedly for gregarines, but none have been found to date. Other 
crabs procured from oyster boats, and which were dredged in the Sound 
and Harbor, have not yielded gregarines. These include Neopanope 
texana sayi, Carcinides maenas, Pagurus bernhardus and Pagurus 
longicarpus; and Chloridella empusa from the mud flats of the Inner 
Harbor. From the south side of the island, the following crabs have 
been examined and none found to be infected with gregarines: Emerita 
talpoida, Callinectes sapidus, Ovalipes ocellatus, Ocypoda albicans, and 
an undetermined species of Orchestia. 

It seems possible that only littoral marine crustacea are infested 
with gregarines and that spores are eaten along with shore vegetation, 


134 THE JOURNAL OF PARASITOLOGY 


grasses and tide water algae, and are swept away by the tides or are 
noninfective when they reach the water. 
A table of measurements, in microns, follows: 


Total length association........... 218 195 150 127 
Primite: 
ZENS ENESDOLOMNE faciiae seis fe sass « 100 85 80 65 
Length protomerite ............ 20 20 14 14 
Length deutomerite ............ 80 65 66 51 
Width protomerite ..........5.. 85 38 30 30 
Width deutomerite ............. 43 48 45 36 
Ratio length protomerite: total 
LENE TMMSPOLONE .. + c/s ses <0 sso 135 1 :4.2 ik ey7/ 1 :4.6 
Ratio width protomerite: width 
GEMEOMERITE™ @ ..c5.0:0655 see - a 1S GIES LZ 
Satellite : 
BenethespOront vaca... eccs = 6 ser 118 110 70 62 
ener protomerite . 55... 6550s 25 14 10 10 
Length deutomerite ............ 83 96 60 52 
Width protomerite ............. 36 39 22 22 
Width deutomerite ............. 36 50 28 30 
Ratio length protomerite: total 
EMetieS POLO ts crac sce mis s -s 5 1:8 1b 37/ 1 :6.2 
Ratio width protomerite: width 
MEULOMERITE <e..c.. cece mac vei) eles iets Dele 


Frenzelina nigrofusca noy. spec. 
Figures 11 to 14 

Two species of fiddler crabs, Uca pugnax and U. pugilator, which 
live together at the roots of the eel grass, were found to be infected 
with a species of gregarine. About 30 per cent. of the hundred or 
more crabs examined were parasitized and the infection was very 
moderate; in only rare instances was the number of parasites present 
greater than fifteen. 

The sporonts were solitary, none being associative as is character- 
istic of this genus. The body is broadly ovoidal, less than twice as 
long as wide (Fig. 11) and is often nearly rectangular in shape with 
rounded corners (Fig. 12). Sporonts average 100y in length and 65n 
in width. The protomerite is hemispherical and very slightly con- 
stricted at the septum; it is about one third the length of the whole 
sporont. There is a minute papilla at the anterior end; this papilla is 
large and conspicuous in the trophozoite (Fig. 13). The deutomerite | 
is of approximately the same width as the protomerite and twice as 
long. It is very broadly rounded or often flattened posteriorly. 

The endocyte is very dense and appears dark brown or black in 
transmitted light. It is but little less dense in the protomerite than in 
the deutomerite and in starved animals becomes tan in color. The 
nucleus is not visible in the live sporonts. The sarcocyte is relatively 
thick, especially over the anterior end of the protomerite. The nucleus 


WATSON—GREGARINES FROM MARINE CRUSTACEA — 135 


is small and spherical and contains one or two minute karyosomes. 
Uniform gliding movement was observed at a relatively slow rate. 
The cysts are very dense and are spherical. Spores were not seen. 

This species is placed in the genus Frenzelina for two reasons: 
(1) the protomerite possesses at its apex a small papillated and con- 
spicuously colored disk, this papilla being well developed in the tropho- 
zoite, (2) the gregarine infects the intestine of a marine crustacean. 
While no associative sporonts were seen, the species is very probably 
associative from its affinities. 

A table of measurements of a few specimens is appended here, all 
dimensions being given in microns: 


Total length sporont.......... 72 82 100 120 125 
Length protomerite .......... 20 22 22 31 31 
Length deutomerite .......... 52 60 78 89 94 
Width protomerite ........... 39 40 70 50 75 
Width deutomerite ........... 37 40 75 75 65 
Ratio length protomerite: total 

MOL S tlie eee scccta sre seta oe orate eNO oor wigs © I 4 1:4 
Ratio width protomerite : width 

GEUTOMENITS 1 he esa. cen semen isi ies | ibsal 13 Lore ai DA 
Diameter nucleus. ............ 1 ilps 125 

SUM MARY 


1. Three new Gregarine parasites of the genus Frenzelina are 
described from marine Crustacea. 

2. Parasites belonging to this genus have not hitherto been reported 
from the United States. 

3. A new definitive character for this genus has been determined 
upon sectioning the host intestine, namely, the fact that the para- 
sites are intercellular. 


REFERENCE CITED 


Léger, L., and Duboseq, O. 1909. Etudes sur la sexualité chez les Grégarines. 
Arch. Prot., 17: 19-134. 


136 THE JOURNAL OF PARASITOLOGY 


EXPLANATION OF PLATE 


Frenzelina delphinia nov. spec. 


1. An association of two sporonts from lumen of intestine of Talorchestia 
longicornis; a, protomerite of sporont, b, deutomerite, c, primite, d, satellite. 

2. Another association with satellite much younger than primite. 

3. Cross section of portion of intestine of Talorchestia longicornis showing 
intercellular development of parasite; a, oblique section of trophozoite embedded 
in epithelium; b, c, sections of sporonts lying free in lumen but near the walls 
and surrounded by mucus. 

4. Interlocking device by which satellite of association is attached to primite. 

5. Association revolving in an early stage of cyst formation. 

6. Cross section of portion of host intestinal epithelium showing embedded 
trophozoite in longitudinal section. 

7. Cyst soon after completion, showing transparent cyst wall and two sporonts 
still distinctly outlined. 


Frenzelina olivia nov. spec. 


8. Mature association from lumen of intestine of Libinia dubia. 

9. Immature sporont from same location. 

10. Young sporont free in intestinal lumen, showing papilla at anterior end of 
protomerite. Magnification greater than in Figures 8 and 9. 


Frenselina mgrofusca nov. spec. 


11, 12. Adult sporonts from lumen of intestine of Uca pugnax. 

13. Trophozoite from lumen of host, showing papilla. 

14. Sporonts with posterior half of body contracted, indicating that consid- 
erable movement of the body is possible. 


PLATE 


eet EET ES 


THE PAJAROELLO TICK (ORNITHODORUS CORIACEUS 
KOCH) 


WITH SPECIAL REFERENCE TO LIFE HISTORY AND BITING HABITS 


WILLIAM B. HeERMs 


University of California 


For several years previous to beginning his observations on this 
species, the writer has listened to many harrowing tales about the 
Pajaroello. No one seemed to know exactly what it was and no one 
seemed to have collected specimens so as to make accurate identification 
possible in so far as the writer knew at the time. Complaints came 
almost exclusively from the more mountainous portions of Santa Clara 
and San Benito Counties (California). Natives, principally Mexicans, 
in the vicinity of Mt. Hamilton fear this parasite more than they do the 
rattlesnake, and tell weird tales of this or that man having lost an arm 
or leg, and in one instance even death having ensued, as the result of 
a bite by the Pajaroello. There seems to be a superstition in that region 
that three bites will result in certain death. The stories all agree in 
the essential detail that the bite results in an irritating lesion which 
is slow to heal and often leaves an ugly deep scar. Several persons 
also informed the writer that the Pajaroello occurred in certain moun- 
tainous portions of Mexico. It was not, however, until August, 1913, 
that living specimens came to hand, taken in Santa Clara County in 
the vicinity of Mt. Hamilton. These were identified as Ornithodorus 
coriaceus Koch, described in 1844 from a single female specimen from 
Mexico. A translation by Nuttall of the original description is as 
follows: 


“Shaped like the sole of a shoe, thick margined, roughly shagreened, yel- 
lowish earthy color, spotted rusty red, legs toothed dorsally. Length 9.3 mm. 
Body about twice as long as wide, width fairly uniform, indented on the sides, 
pointed above the mouthparts, rounded posteriorly, a thick turned-up border 
all around; the whole surface above and below thickly granulated like fish 
skin (shagreen), the granules flat above, consequently, the whole leathery, on 
the back unequal folds and grooves. Beneath in the front of the body a deep 
groove running to the stigmata and on the inner protrusion the rather large 
round quite clearly marked eyes. The coxae gradually thicken toward the 
distal extremity and are somewhat bent; the other articles somewhat com- 
pressed and clearly notched or round-toothed. The whole surface, above and 
below, dirty yellowish earthy color, rusty red spots irregularly distributed 
throughout. Capitulum and palps light yellow. Legs gray-brown. Female. 
Male: unknown. Habitat: Mexico.” 


138 THE JOURNAL OF PARASITOLOGY 


From either specimens received or reliable information at hand 
it now seems evident that this species occurs in the more mountainous 
portions of the following counties in California, namely, San Benito, 
Santa Clara, Stanislaus, Monterey and Santa Barbara, probably also 
Los Angeles and San Diego, thus connecting up with Mexico, which 
is probably the original habitat. The tick is most commonly found 
among the dry leaves beneath live oak trees where cattle are accus- 
tomed to lie in the shade. Most cases of tick bite caused by this species 
have occurred while sitting or lying down in such situations. 

This species of tick is a typical representative of the genus Ormi- 
thodorus of the family Argasidae and is superficially not greatly unlike 
the relapsing fever tick of Africa, namely, Ormithodorus moubata 
Murray. 

Since August, 1913, the complete life history of this tick has been 
worked out and much information has been gained relative to its 
habits and venomous properties. In this work the writer has been 
greatly assisted by several of his advanced students, notably Mr. W. L. 
Chandler, who has undertaken an exhaustive study of this species. 


LIFE HISTORY 


Six adults and half-grown specimens, males and females, were 
secured during the month of August, 1913. Of these Tick No. 6, a 
fully grown female, engorged on blood from a Rhesus monkey, 
November, 1913, deposited a lot of eggs Feb. 13, 1914, and continued 
to lay eggs at intervals during the rest of that season. Various experi- 
ments were performed with the ova and the several larvae resulting 
from the first laying, mainly for the purpose of determining the best 
method of procedure. From the second laying of this tick we secured 
our first complete life history data as follows: March 9, Tick No. 6 
deposited 323 ova, hatched March 31; giving an incubation period of 
about 21 days at an incubator temperature averaging 26.3 C. (varia- 
tion + 1C.). The larvae were placed on the ear of a rabbit May 2 and 
among others one was recovered fully engorged May 11, and given 
the number 18. The first moult occurred May 21, giving about 51 days 
for the larval stage in this instance. The second moult without a sec- 
ond engorgement took place June 15. The nymph became fully 
engorged in about twenty minutes on July 2, the third moult occurring 
August 12. Becoming fully engorged again October 11, the fourth 
moult took place December 23. Engorging again Jan. 16, 1915, the 
fifth moult took place March 9 and the sexually differentiated tick 
(a female) appeared. March 27 it became fully engorged on a 
mouse and was placed with male No. 3 on April 16, copulation 
taking place April 17. The first laying consisting of 428 eggs 
took place June 10, 1915. Thus the egg to egg period in this 


HERMS—THE PAJAROELLO TICK 139 


invidual covered exactly fifteen months. This time can be reduced 
very considerably by applying the ticks to a suitable host animal 
at shorter intervals, indeed we have one record of a male in which sex- 
ual differentiation was accomplished in 159 days, as against 343 days in 
Tick No. 18, a female. Under natural conditions it seems quite prob- 
able that there is one generation each year and that two years may be 
necessary in many instances. 

Although the incubation period at a given sustained temperature 
suffers little variation, e.g. at 26 C. it is 21 days, the length of time 
required for the other stages varies considerably, depending on the 
presence of a host mainly. 

The minimum length of the larval period was found to be 19 days. 
The number of moults varies from four to seven. 


Fig. 1—The Pajarocllo tick, Ornithodorus coriaceace Koch. Dorsal, left; 
ventral, right. 


The length of time a female may remain fertile without further 
copulation is at least two years as shown by the fact that Tick No. 6 
received as a fully grown individual August, 1913, and not thereafter 
placed with a male, deposited eggs during the summer of 1914 and 
1915. The total number of eggs deposited in one season by Tick No. 
6 was 1,158, there being seven separate layings. The maximum num- 
ber of eggs deposited in a single lot for the year 1914 by Tick No. 6 
was 323. This same tick, however, deposited 802 eggs April 26, 1915, 
her tenth laying in captivity, and her daughter, Tick No. 18, deposited 
428 in her first laying. 

We have experienced no little difficulty in rearing this species of 
tick. However, the ear of a rabbit is best suited for feeding the larval 
ticks ; later stages are best fed by placing the ticks either on a rabbit 
or on a mouse, holding these with the hands until the ticks have become 
fully engorged and drop off, this process requiring from 15 to 30 
minutes. 


140 THE JOURNAL OF PARASITOLOGY 


BITING HABITS AND VENOMOUS NATURE 

Mr. W. L. Chandler, a graduate student in the University of Cali- 
fornia, formerly with the United States Public Health Service, has 
given the writer an accurate account of two bites which he suffered 
while stationed in the San Antone Valley (California). The first bite 
was received July 2, 1912. He experienced a sharp pain on the left 
arm and upon rolling up his sleeve discovered a large tick, partly 
engorged, attached to the upper arm in front. He dislodged the 
tick and sucked the lesion. The lesion when first discovered showed a 
small dark purple ring surrounding a bright red spot, the point of 
attachment. The discoloration disappeared in a short time but the arm 
was “highly irritable for two or three days and at the point of attach- 
ment a minute clear scab formed.” The tick proved to bea _ pajaroello. 

The second bite took place July 16 while seated in a thicket of 
willows (the first bite took place while riding over a brush grown hill), 
and in this case the sharp pain involved the left leg. An almost fully 
engorged tick, again a pajaroello, measuring about three-fourths of an 
inch in length and about one-half inch in width was removed from 
just above the shin. Once more a bright red spot was visible at the 
point of attachment surrounded by an irregular purple ring about three- 
fourths of an inch in diameter. In about half an hour the leg began to 
swell in the vicinity of the lesion and in about three hours the entire 
lower leg was tremendously swollen. The coloration about the point 
of attachment had widened considerably, was puffy and a clear lymph 
exuded from the lesion. The young man lanced the leg causing the 
blood to flow freely and treated the wound with crystals of potassium 
permanganate, binding the leg with cotton and gauze. During the fol- 
lowing night he reports experiencing a general disagreeable feeling, the 
entire lower leg being “irritable and numb.” On the following day the 
bite on the arm became irritable again, and was treated as had been the 
leg, fearing bad results. For several weeks both lesions exuded a clear 
lymph from beneath an “oily looking, transparent, red mottled scab, 
which remained in evidence for two or three months.” 

Chandler reported these ticks very numerous in some localities, 
having counted as high as six within half an hour crawling over a sad- 
dle blanket placed on the ground. Their presence and number seemed 
to be determined by the presence of cattle, although ticks were found 
where there were no cattle but in places which where evidently favorite 
haunts of wild-animals. 


EXPERIMENTS WITH THE PAJAROELLO 

On monkeys: A number of specimens of Ornithodorus coriaceus 
were collected in the San Antone Valley and at Newman, California, 
for purposes of experimentation and study of life history. In coopera- 


HERMS—THE PAJAROELLO TICK 141 


tion with Dr. W. A. Sawyer and Messrs. S. W. Newman and W. L. 
Chandler, the writer conducted a number of experiments particularly 
with reference to the bite. In one of these experiments a mature female 
tick was permitted to bite a nearly full grown monkey (Macacus 
rhesus) twice within an interval of sixteen days intervening between 
the two bites. The tick was applied at 9:42 a. m. Dec. 10, 1913, and 
began sucking blood at 9:43, one minute later, becoming engorged and 
falling off at 10:21 a. m., a period of 38 minutes. At 10:30, a few 
minutes after the tick dropped off, there appeared a deep red 
hemorrhagic area 2 mm. in diameter at the point of biting with a some- 
what lighter area 10 mm. in diameter surrounding the central area. 
At 10:47 there was a black spot at the point of bite 1.5 mm. in diameter. 
the inner red hemorrhagic area measuring 4 mm., with a yellowish 
white area surrounding this 8 by 6 mm., and an outer petechial area 
15 by 13 mm. No general symptoms were noted. The lesion reached 
its greatest expanse the following day when the following measure- 
ments were taken: dark purple spot 2 mm. in diameter (a very dark 
red scab) ; the inner red area 6 by 5 mm., the yellowish white area 20 
by 12 mm., the outer area 48 by 23 mm. and fading. The yellowish 
white area including bite was slightly swollen. By December 14, te. 
four days after the bite was received, the ecchymosis had entirely dis- 
appeared; by December 16, six days after the bite, the lesion was 
entirely gone but for a slight pigmentation, a thickened reddish area 
measuring 5 by 3 mm. and a small scab 2 mm. in diameter. 

The monkey remained normal throughout the experiment as regards 
temperature, weight, blood count and general condition. 

The second bite was received by the same animal on December 26, 
the tick being applied at 9:43 a.m., taking hold at 9:44 a.m. and 
dropping off fully engorged at 9:55 a.m., requiring but 11 minutes to 
engorge. The history of the second bite follows that of the first very 
closely, except for the extent of the lesion which was greater, i.e. 70 by 
30 mm. In order to note any manifestation on the part of the first 
lesion, the second bite was located near the opposite nipple. No change 
was observed. The lesion produced by the second bite had disappeared 
by December 31, i.e. five days after the bite, except for a slight thicken- 
ing 3 mm. in diameter and a slight white scale at the center. Again the 
monkey had remained normal, except for a slight increase in the count 
of white blood corpuscles which rose from 7,400 at the time of the bite 
to 13,900 by noon of the same day, going down again to 7,300 by 5 p.m. 

Both nymphs and adults readily attach to man, monkey, rabbit and 
mouse ; and become fully engorged in from 15 to 30 minutes, depending 
on the number and length of rests in the rhythmic motion of the basis 
capituli. If dislodged while engorging, they, like the larvae, refuse to 
reattach immediately. 


142 THE JOURNAL OF PARASITOLOGY 


On rabbits: The bite on a rabbit’s ear leaves a comparatively large, 
thick, purple nodule, and is accompanied by a more or less hemorrhagic 
condition of the entire ear tissue. The bite has no apparent systemic 
effect on the rabbit, and the lesion heals within two or three weeks. 

On mouse: When a tick was applied to the body of a mouse for the 
first time, the mouse showed no apparent uneasiness until the tick 
attempted to withdraw its head. Beginning immediately after the tick 
dropped from the mouse there occurred a small swelling which exuded 
lymph, rapidly grew in proportions and was accompanied by marked 
ecchymosis. In 2 minutes after the tick had dislodged itself the swell- 
ing had increased its area 0.3 by 1 cm., in 30 minutes 0.5 by 2 cm., and 
in one hour 0.5 by 2.5 cm. After 24 hours the swelling had become 
reduced, except in the vicinity of the bite, and fully one half of the 
mouse was dark blue. But slight systemic disturbances were noticeable 
in the mouse, and it rapidly recovered. The same mouse was used as 
a host for other ticks, and each succeeding bite produced less and less 
noticeable results, until finally only very slight lesions were produced. 


SUMMARY 


The venomous Pajaroello Tick (“Pajahuello” according to Banks) 
is a native of Mexico, but is now known to occur in the more mountain- 
ous coastal counties of California as far north as Santa Clara within 
100 miles of San Francisco. 

The breeding habits, metamorphosis and life history have been 
carefully observed in the Parasitology Laboratory of the University 
of California. . Records of life history in individual cases (i. e., 
from egg to sexual maturity) show 159 days for the male and 343 days 
for the female at a temperature averaging 26.3 C. (variation + 1 C.). 
The venomous nature of the bite as affecting man, monkey, rabbit, and 
mouse is described. 


NOTE ON THE ETIOLOGY OF VERRUGA AS DEDUCED 
FROM A STUDY OF THE ASEXUAL STAGES 
OF BARTONELLA 


Cuartes H. T. TowNsenp 


Bureau of Entomology, U. S. Department of Agriculture 


Dec. 4, 1915, the writer presented before the Biological Society of 
Washington a paper on the identification of the asexual stages of 
Bartonella bacilliformis, the causative organism of verruga, which was 
published in the Dec. 19, 1915, issue of the Journal of the Washington 
Academy of Sciences. 

Jan. 7, 1916, he presented the same subject before Section VIII of 
the Second Pan-American Scientific Congress, during which presenta- 
tion he announced certain points additional to those previously 
announced. This second paper will be published in due course in the 
Proceedings of the Congress, but it is desirable to place on record at 
once the additional points announced therein. They are as follows, 
and should be added to the paper published Dec. 19, 1915: 

The toxin resulting from the extensive asexual multiplication of 
Bartonella in the vascular endothelial cells of the subcutaneous tissues 
is liberated in quantity into the blood, causing the rise of temperature 
which marks the fever stage of verruga, the anemia following through 
hemolysis. 

The proliferation of vascular endothelial cells incited by this toxin 
not only imprisons the toxin itself, thus arresting the hemolysis, but 
also prevents the erythrocytes from coming into direct contact with 
endothelial cells containing merozoites of Bartonella, thus cutting short 
the infection of the erythrocytes. As the natural result, the fever and 
anemia both subside, and the gametes of Bartonella are no longer to 
be found in the peripheral blood. 

The infected endothelial cell, in situ in the capillary wall, is posi- 
tively chemotropic for uninfected freshly oxygenated erythrocytes, 
attracting and holding them in contact with itself until transfer of a 
certain number of merozoites of Bartonella has been effected, the 
presence of which reduces the oxygen tension in the substance of the 
erythrocytes, thereby transforming their tropic qualities, the sufficiently 
infected erythrocytes being set free through negative chemotropism. 

The localized proliferation of vascular cells following verruga 
eruption-tissue inoculations is not due to any new activity of a living 
organism or virus. The reason why Drs. Strong et al. were unable 
to obtain proliferation lesions by injection of a filtrate from these tis- 


144 THE JOURNAL OF PARASITOLOGY 


sues is at once apparent; the proliferated vascular cells can not pass 
the filter. Their inoculation of these tissues upon the rabbit’s cornea 
produced no lesion because the cornea possesses no vascular cells. 
Their attempts to cultivate the supposed virus in these tissues resulted 
in failure because the tissues evidently do not contain a living virus. 
(The term virus is used in the common acceptation of organisms that 
pass the filter.) 

Drs. Strong et al. succeeded in demonstrating the presence in ver- 
ruga eruption tissues of a hemolysin which is active in relatively high 
dilutions, and whose discovery is very much to the point in this par- 
ticular connection. This hemolysin is quite certainly the toxic by-prod- 
uct of the reproductive activity of Bartonella in the subcutaneous tis- 
sues. It is the specific cause of the anemia of the fever stage of 
verruga. Further, it is the agent which directly incites the prolifera- 
tion of the vascular cells, thereby causing the eruption lesions. In 
other words, this toxin is able to destroy such delicate structures as 
the erythrocytes; is able to irritate the more resistant vascular cells 
sufficiently to cause them to proliferate; but is unable to produce any 
effect on such highly resistant structures as the connective-tissue cells 
composing the cornea. Its proliferative action on vascular cells con- 
tinues through many successive series of inoculations, but finally 
becomes attenuated and no longer effective. 

In verruga cases, when the correspondence in intensity of fever and 
visible eruption is not well marked, it is practically certain that infection 
of the internal organs has become proportionately greater, resulting in 
an increased internal eruption. 

Notwithstanding all criticisms that may be put forward, the writer 
is content to rest his thesis upon the evidence presented, and invites 
a comparison of the published figures and descriptions of the vascular 
cell inclusions of the fever and eruptive stages of verruga. He has 
verified the findings by similar inclusions in his own verruga sections 
and smears. There are many additional details apparent at the pres- 
ent writing which have not yet been entered into, but they are unneces- 
sary to the main demonstration ; it is only necessary to say that all the 
details fit in perfectly with the identification that has been given of the 
asexual stages of Bartonella, thus clinching the interpretation of these 
as presented by the writer. 


A NEW INFUSORIAN PARASITE IN SAND FLEAS * 


MINNIE E. WaTSON 
Oyster Bay, Long Island, N. Y. 


While examining some sand fleas on the shores of Long Island 
Sound for gregarine parasites, the writer found two hosts infected 
with an infusorian parasite; one of the hosts being the common small 
flea, Orchestia agilis, the other the larger Talorchestia longicornis. 
The two infected hosts were found in the same habitat and on the 
same day. The infusoria were found in only these two instances out 
of about 300 fleas examined for gregarines. The parasites infect the 
alimentary tract of the host in great numbers, several hundred being 
present in each flea. 

In the vegetative stage, the animals are broadly ovoidal, tapering 
slightly at one end (Fig. 1). The macronucleus is very large, and 
ellipsoidal in shape; it is granular and homogeneous and does not 


EXPLANATION OF FIGURE 


1. Balantidium orchestium, vegetative individual. Og, oral groove; oe, 
esophagus; ma, macronucleus; mi, micronucleus; fv, food vacuole; cv, con- 
tractile vacuole. 

2, 3, 4. Three stages in binary fission. 

5. Conjugation. 


stain deeply with Ehrlich’s hematoxylin and acetocarmin. The micro- 
nucleus is small and deep staining, and lies contiguous to the macro- 
nucleus. The apical or subapical oral groove is small and inconspicu- 
ous; it leads to a short slender esophagus. Small contractile vacuoles 
were observed, not more than one being seen in a single individual. 
Many small food vacuoles were often present. Cilia of uniform length 


* Contributions from the Zoological Laboratory of the University of Illinois, 
under the direction of Henry B. Ward, No. 61. Also from the Biological 
Laboratory, Cold Spring Harbor. 


146 THE JOURNAL OF PARASITOLOGY 


cover the body, those in the oral groove being slightly the longer. 
The size of vegetative forms ranges from 300 to 360 microns by 180 
to 220 microns. 

In individuals which are ready for asexual reproduction, the body 
becomes elongate and tapers at the ends; the macronucleus becomes 
longer and constricted centrally, at the same time contracting in vol- 
ume. It also stains more deeply than does that of the vegetative animal 
(Figs. 2,3, and 4). When transverse fission is complete, the two nuclei 
attain their normal vegetative form and density. Conjugation was 
noted in one instance. 

The parasite may be classified as follows: 

1. Order Heterotricha: zone of large cilia leading to mouth. 

2. Suborder Polytricha: body covered with an even coat of cilia. 

3. Family Bursaridae: peristome broad, body broad and large. 

4. Genus Balantidium: parasitic, inhabiting the alimentary tract of 
the host, body ovoidal or ellipsoidal, blunt at ends in vegetative stage, 
macronucleus ellipsoidal. 

5. The parasite is closely allied to B. coli Stein and B. elongatum 
Stein; it differs from them in size of the body and in relative size of 
the nucleus. No previous record of this form has been found. 

I wish therefore to designate this species Balantidium orchestium. 


Urbana, Ill., June, 1915. 


REFERENCES CITED 
Doflein, F. 1911. Lehrbuch der Protozoenkunde. Jena, 1043 pp. 
Minchin, E. A. 1912. An Introduction to the Study of the Protozoa. Lon- 
don, 520«pp. 


Report oF First Expepition to SoutH AMeEricA. Harvard School of Tropical 
Medicine. Richard P. Strong, Ernest E. Tyzzer, Charles T. Brues, A. W. 
Sellards, J. C. Gastiaburu. Cambridge. Harvard University Press, 1915. 
220 pages. 48 plates. 


This very excellent report deals with the investigations of the first expe- 
dition sent out by the Harvard School of Tropical Medicine, to study certain 
forms of tropical disease occurring in South America. The report treats of the 
sanitary conditions of the places visited, namely, Kingston, Jamaica, Colon, 
Panama, Buenaventura, Guyaquil, Callao, Lima, and certain mountain towns 
in the interior of Peru. 

While at Guayaquil yellow fever was very prevalent and many cases of 
this disease were studied. Especial attention was paid to the study of the 
blood, owing to the reports of Seidlin regarding the presence of a protozoon 
which he has named Paraplasma flavigenum and which he regards as the cause 
of the disease. As a result of their investigation of the blood in yellow fever, 
these investigators say: “We were unable, however, to detect any bodies 
which suggested a parasitic nature in the blood in this disease.” 

The larger portion of the report deals with the investigations of oroya fever 
and verruga peruviana and as the results of their investigations the authors 
have determined, apparently without question, that these conditions are dis- 
tinct diseases. Verruga peruviana is due to a virus which may be transmitted 
to animals by direct inoculation, the nature of which is still unknown, while 
oroya fever is due to a parasite of the red blood corpuscle, which cannot be 
transmitted to animals. The parasite concerned in the etiology of this dis- 
ease is of interest to protozoologists because it apparently belongs to a new 
genus which Strong has called Bartonella, naming the genus after Barton, who 
in 1905 first described the bodies within the red corpuscles. The specific organ- 
ism causing oroya fever has been named Bartonella bacilliformis. Multiple 
infection of the red cells is common in severe infections, as many as ten of the 
parasites being present in one cell. Morphologically the parasites occur in the 
form of minute rods resembling a bacillus, measuring from 1 to 2 microns in 
length and rounded or oval forms, measuring from 0.3 to 1 micron in diameter. 
Both rods and rounded forms contain, in stained preparations, granules which 
may be interpreted as chromatin. The investigators were unable to infect either 
monkeys or rabbits with this parasite. 

While unable to find any parasite in the cases of verruga peruviana studied, 
the authors were able to produce very typical lesions in both monkeys and 
rabbits, thus demonstrating the distinct nature of the two diseases. They were 
unable to definitely decide whether the virus of verruga is filterable, but believe 
that ultimately it will be shown to be so. As regards the transmission of the 
two diseases the authors were unable to determine in what manner this occurred 
but lean to the opinion that some arthropod is concerned. 

Uta, a disease of the skin, occurring in Peru, was also studied and the 
authors were able to demonstrate that this disease is due to a species of 
Leishmania. As regards the species they say: “For the present at least, from 
the evidence available we do not feel justified in creating a new species for the 
parasite discovered by us as the etiological factor of uta.” A wise decision in 
view of the present unsatisfactory classification of the Leishmanias. 

The report also contains a description of a new Linguatulid parasitic in the 
crocodile, in Equador, which Wheeler has named Poracephalus crocodili; a 
discussion by Brues of the flies of the family Phoridea obtained by the expe- 
dition, and some notes on Peruvian mosquitoes and mosquito literature, by Knab. 


148 THE JOURNAL OF PARASITOLOGY 


The book is beautifully printed and illustrated and forms an excellent 
example of the good work that may be accomplished by medical expeditions. 
The authors are to be congratulated on the success of their labors for they 
have contributed a most interesting and valuable addition to the literature and 
knowledge of tropical medicine. 


MepicaL ZooLocy IN At present marked attention is being paid to 
parasitology as is evidenced by recent publications noteworthy in form and 
character. This movement has found its expression both in monographic articles 
accorded a place in general medical literature and in a special periodical devoted 
to the subject. Brief comments on these items will be of service to North 
American parasitologists\ who may not have seen the original publications as yet. 

Archivos Brasileiros de Medecina (Anno ivy, nos. 1, 2, 3) contains an extended 
discussion of hookworm and hookworm disease covering seventeen separate 
contributions on all phases of the subject, illustrated by nine full-page plates. 
The work is complete and scholarly in treatment, giving much that is not 
available elsewhere. Of especial value is the critical bibliography on ancylos- 
tomiasis in Brazil. 

Memorias do Instituto Oswaldo Cruz, published at Rio de Janeiro, which 
reached its seventh volume in 1915, includes original articles on protozoology, 
helminthology, and medical zoology generally. The text is usually printed in 
Portuguese and French or German in parallel columns. The journal is illus- 
trated by full page plates in heliotype and lithography in colors, which are very 
sucessful and deserve high praise. Most articles concern those elements of the 
fauna of Brazil which are related to the cause and dissemination of disease. A 
recent number contains also a sketch of Professor von Prowazek accompanied 
by a fine portrait. 


ARCHIVES DE ParasitoLocy.—Publiées par Raphael Blanchard, Professeur a 
la Faculté de Médecine de Paris, Member de l’Académie de Médecine. 

Les Archives de Parasitologie, imprimees a Lille (Nord), ont di suspendre 
leur publication, par suite de l’occupation de la ville de Lille par les armées 
allemandes. 

Le fascicule du tome XVI porte la date du Ist aout 1914. Il était remis 
au chemin de fer, quand le guerre a éclaté. Il a di étre retiré et, dupuis lors, 
n’ a pu étre distribué. 

Voici son sommaire: 

P. S. de Macaruaes, Notes d’helminthologie brésilienne, dixieme série (avec 
7 figures dans le texte), page 481. 

R. Brancuarp, Notices biographiques, XXIV. Louis-Daniel BEAUPERTHUY, 
1807-1871 (avec 2 figures et 7 fac-simile dans le texte), 503. 

G. R. Branc, Sur quelques espéces du genre Diplotriena Railliet et Henry 
(avec 10 fig. dans le texte), 546. 

P. Mora, Cestodes Avium. Contributo alla fauna elmintologica sarda (pl 
TX), D57- 

Les Parasites et les maladies parasitaires dans l’historie, la poésie et l'art, 
579-637. 

Notes et Informations, 638. 

Table des matiéres, 639-640. 

Le premier fascicule du tome XVII était en bonne voie d’impression; les 
cing premiéres feuilles étaient déja tirées,au moment de la déclaration de guerre. 

Les Archives ne pourront reprendre leur publication réguli¢re qu’aprés la 
cessation des hostilités. Elles paraitront alors par fascicules moins gros et 
plus rapprochés. 


The Journal of Parasitology 


Volume 2 JUNE, 1916 Number 4 


THE SIGNIFICANCE OF CERTAIN NATURAL FLAGEL- 
LATES. OF INSECTS IN THE EVOLUTION OF 
DISEASE IN VERTEBRATES 


H. B. FANTHAM 


Lecturer in Parasitology, Liverpool School of Tropical Medicine, 
and Christ’s College, Cambridge 


AND 


ANNIE PoRTER 
Beit Memorial Research Fellow, Quick Laboratory, Cambridge 


INTRODUCTION 


During the last few years, considerable attention has been given 
to the role of insects in the spread of disease. Much work has been 
done in elucidating life-histories, more particularly of the parasitic 
flagellates peculiar to insects and having no apparent connection with 
vertebrate maladies. Many flagellates that are seemingly limited to 
insects, however, are not so innocuous to vertebrates as they appear 
at first sight. The introduction of certain of the parasitic Mastigo- 
phora, notably members of the genera Herpetomonas and Crithidia, 
into vertebrates by the latter swallowing the infected insects or by 
forms of the parasite entering the vertebrate host by way of wounds 
or abrasions of the skin, has been shown to result in pathogenic effects 
to the said hosts. Laveran and Franchini have demonstrated the exis- 
tence of this capacity for exercising latent pathogenicity by infecting 
dogs with the flagellates of dog fleas, and rats and mice with the flagel- 
lates of the fleas infesting these rodents. The present authors, working 
quite independently of Laveran and Franchini, have conducted a series 
of experiments extending over some six years on the possible patho- 
genic effects that accrue when certain flagellates of insects reach either 
associated or unassociated hosts. We have considered more especially 
the evolution of disease as exemplified by flagellates that have induced 
a flagellosis in vertebrates, remembering that, at any rate in some cases, 
parallel conditions prevail in Nature and in our experiments. By the 
introduction of certain herpetomonads normally parasitic in insects into 
vertebrates, a condition resembling leishmaniasis or kala-azar in man 


150 THE JOURNAL OF PARASITOLOGY 


has been produced, the symptoms of the disease and the morphology of 
the parasites found therein showing that here are, at least, examples 
of “leishmaniasis in the making.” 

The evolution of the parasitic habit with the development of patho- 
genicity as the result of change of habitat is no new phenomenon to 
those who have carefully studied the comparative morphology and life 
histories of various pathogenic Protozoa. Change of habitat has fre- 
quently led to great alteration in the mode of life of an organism. 
Thus, when a herpetomonad has so adapted itself that it is capable of 
living and propagating in a vertebrate, as has probably happened in the 
case of Leishmania, an originally monogenetic parasite by the exertion 
of its capacity for plasticity becomes digenetic. The various herpeto- 
moniases, of which the leishmaniases are a special section, are probably 
the result of the introduction of insect or other invertebrate flagellates 
into the vertebrate hosts. In the case of acute forms of disease, the 
excitants show less perfect power of adaptation to the new environ- 
ment than do the parasites that induce the chronic type of malady. 
From the point of view of the parasite, the maintenance of the life of 
the host is an economic desideratum, the prolongation of the active life 
of the invader depending in part on the longevity of the host. The 
newer a parasite is to the animal harboring it, the less it is in harmony 
with its environment. The consequence is that its discord with the host 
is manifested by pathogenic effects and the latter animal succumbs. 
Chronic maladies are usually correlated with greater powers of adapta- 
tion of the parasite to its host, with the period that has elapsed since 
the original introduction of the parasite to the host, and with the rela- 
tive resisting powers of the host to the specific action of the parasite. 

Certain trypanosomes appear to have developed from the flagellates 
of certain insects (for example Drosophilia), which parasites in turn 
seem to have been derived from free-living forms. As free-living 
organisms, their power for harm appears to be negative. If they 
become saprophytic, their capacity for developing noxious qualities is 
increased. When the parasitic habit of life, such as bloodsucking, is 
established in an insect, the power for injury possessed by its contained 
flagellates is greatly extended. Finally when the insect flagellate 
reaches the higher vertebrate the recapitulative effects of its evolution 
manifest themselves with cumulative results and the parasite is defi- 
nitely pathogenic. The scale of evolution thus outlined is exhibited by 
the Trypanosomidae, some of whose members, the trypanosomes and 
herpetomonads (Leishmania) of vertebrates are notably pathogenic. 
While they are in the insect host, to which they have adapted them- 
selves, they are relatively innocuous. 

Yet other Protozoa afford studies in the evolution of pathogenicity. 
It must suffice to mention the case of the malarial parasites. The 


FANTHAM-PORTER—EVOLUTION OF DISEASE 151 


morphology of these organisms shows that they are closely allied to the 
Coccidia, and there is little doubt that-the malaria excitants were 
originally Coccidia of insects, that with change of habitat developed 
increased powers of adaptation to life in vertebrates, and, at the same 
time, increased in pathogenicity towards the new host. 

The evolution of disease in the past is presented to us by the 
example of the malarial parasites. Disease in the making is manifested 
to mankind today in the case of the herpetomonads more especially. 
Preventive measures gain in efficacy as the natural modes of infection 
and possible sources of disease excitants are considered. In this con- 
nection, the experiments that we have undertaken may be of service 
as indicating possible sources of disease hitherto unsuspected. 


MATERIAL AND METHODS 


The materials used in this research consisted of various flagellates 
found in insects on the one hand, and of representatives of each of the 
great phyla of European vertebrates on the other. 

The flagellates used in our experiments included members of the 
genera Herpetomonas and Crithidia. They comprised Herpetomonas 
jaculum Léger, parasitic in the gut of the Hemipteran, Nepa cinera; 
H. stratiomyiae Fanham and Porter, from the intestine of the Dipteran, 
Stratiomyia chameleon; H. pediculi Fanham, from the alimentary tract 
of Pediculus vestimenti,; H. culicts Novy and MacNeal, from the larvae 
and adults of the gnat, Culex pipiens; and Crithidia gerridis Patton, 
parasitic normally in the alimentary tract of the Hemipteran, Gerris 
paludum. 

The vertebrate hosts included representatives of the Pisces (Gaste- 
rosteus aculeatus), Amphibia (Rana temporaria, Bufo vulgaris and 
Molge vulgaris), Reptilia (Lacerta vivipara and Tropidonotus natrix), 
Aves (Serinus canarius, Passer domesticus and Chelidon urbica) and 
Mammalia (Canis familiaris and Mus musculus). 

The insect flagellates were introduced into their respective verte- 
brate hosts either by inoculation or by feeding. In the latter case, the 
host was fed with the infected insects, or with the intestines of the 
insects, or with food contaminated with the feces of the insects con- 
taining the resistant (nonflagellate or postflagellate) stages of the 
Flagellata concerned. After the infective feed or feeds, ordinary food 
was given. The hosts were examined for blood parasites and for ecto- 
parasites prior to use, and were found free from both classes of infesta- 
tion. 

Blood films were made periodically during the life of the infected 
animals, and smears of the internal organs were prepared at autopsy. 
Some preparations were fixed while moist with osmic vapor followed 
by absolute alcohol, while others were fixed wet with Bouin’s fluid. 


tse THE JOURNAL OF PARASITOLOGY 


Intravitam staining was often employed. For permanent preparations 
Giemsa’s stain, hematoxylin and eosin, iron hematoxylin and occa- 
sionally hematein were employed. 

Control vertebrates were kept in each case. They remained healthy, 
lived longer than the experimental animals and were found to be 
unparasitized when killed. 


EXPERIMENTAL WORK 


The course of our experiments, extending over some years, on the 
introduction of insect flagellates into vertebrates may be gathered from 
the subjoined table. It has not been possible to manage a large number 
of animals satisfactorily at one time, as some have lived for relatively 
long periods. Sometimes it was possible to introduce the more resis- 
tant, encysted, leishmaniform, postflagellate forms of the Herpetomonas 
or Crithidia into the vertebrate. In such cases, it was found that a 
larger proportion of infections ensued than when the preflagellate or 
flagellate forms alone were introduced. 

It is of some interest to note that as a rule the Herpetomonas or 
Crithidia introduced were few in number. The parasites, however, 
adapted themselves to their new surroundings, and both nonflagellate 
and flagellate organisms in all stages of active division have been 
recovered from the infected hosts. In some cases it has been possible 
to observe the completion of the multiplication in fresh preparations of 
the organs of the host. The parasites in the vertebrates are not merely 
conserved. The number of parasites obtained from the vertebrate host 
is superior to that introduced, multiplication of the organisms occurs 
and the host often undergoes pathogenic changes resembling those of 
leishmaniasis, that have frequently resulted in death. The herpet- 
omoniasis induced is therefore regarded as a true parasitic infection, 
both the causal parasites and the maladies having affinities with kala- 
azar. 

The possibility of bacterial contamination of the material used for 
feeding or inoculation has not been overlooked. Heavy bacterial con- 
tamination was exceptional in the insects used for experiment, and here, 
as elsewhere, the flagellates often die out in the presence of many bac- 
teria. Further, in Nature pure cultures of flagellates or other protozoa 
rarely occur, as, for example, is evidenced by the diverse organisms 
found in sores of man in the East and in the mixed flora and fauna of 
the alimentary tract of vertebrates. Also, the insects swallowed by 
such vertebrates as lizards and snakes cannot, of necessity, contain 
pure cultures of flagellates or bacteria—in fact, mixed infections are 
frequent in Nature. 

The tabular summary of our experiments is given on page 154. 


~ 


FANTHAM-PORTER—EVOLUTION OF DISEASE 153 


From the following table, it will be seen that while a number of the 
infections are of the acute type, yet there are others in which the herpet- 
omoniasis induced was of relatively long duration. We may mention 
that when the infection was of the chronic type, the leishmaniform, 
nonflagellate bodies preponderated in the smears of the organs of the 
vertebrate host, while flagellate herpetomonads were more numerous in 
the cases of acute herpetomoniasis. In practically all the animals 
infected experimentally, both nonflagellate and flagellate forms of the 
organism introduced were present, the proportion of each form show- 
ing variation. While these conditions prevailed in our experiments, 
we do not consider that any generalization can yet be made therefrom. 

We would also point out that our experiments show the potential 
danger of many flagellates of insects that may at first sight seem 
unconnected with the vertebrates into which they have been introduced. 
Natural modes of infection, however, occur with a number of exam- 
ples ; thus, the dog may contract infection with herpetomonads by eating 
dog fleas and by ingesting infected flea feces when licking its coat, 
H. jaculum from Nepa cinerea can easily reach the fish and amphibia 
which it attacks and may even reach man by way of the wounds 
inflicted by the raptorial cutting forelimbs used when the insect sucks 
blood. The case of insectivorous birds whose normal food is insects 
is obvious. The experiments, whether between associated or unasso- 
ciated insect flagellates and vertebrates, show “leishmaniasis in the 
making.” 

The chronic infections afford examples of good powers of adapta- 
tion to environment on the part of the parasites. As noted in our 
introduction, it is to the advantage of the newly established organism 
that the life of the host should be prolonged, and thus the continued 
existence of the parasite ensured. The acute cases are marked by the 
rapid development of the flagellate forms of the organisms, and by 
their less perfect adaptation to new surroundings as manifested in their 
pathogenic effects to their new hosts. 

A further point of interest is that when young hosts were used, the 
parasites were more virulent. This is also the case with the parasites 
causing the human disease, Mediterranean kala-azar, which is prevalent 
more especially in children. 


MORPHOLOGY OF THE PARASITES IN THE VERTEBRATE AND 
INVERTEBRATE HOSTS 


The life-history of a herpetomonad in its insect host may be briefly 
outlined as follows: A Herpetomonas is a flagellate possessing also a 
nonflagellate stage in its life-cycle. This nonflagellate form is an ovoid 
or rounded, leishmania-like body containing a nucleus and a blepharo- 
plast. It (Fig. la) may be passed from the host with the feces of the 


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156 THE JOURNAL OF PARASITOLOGY 


latter, and is then surrounded with an outer coat. If the excrement 
containing the nonflagellate—sometimes termed encysted or postflagel- 
late—forms of the herpetomonad is ingested by another insect host, 
these ovoid forms of the parasite have their firm, varnish-like outer 
coat (Fig. 1f) dissolved by the digestive juices of the host and are then 
capable of further development. In this condition, they are often 
termed preflagellate forms (Fig. la). The preflagellate form gradually 
elongates. A flagellum arises near the blepharoplast (Fig. 1c), reaches 
the surface of the body at the anterior end and finally projects as a free 
flagellum. The posterior end also elongates and thus the typical flagel- 
late is produced (Fig. 1d). 

Multiplication of the flagellate by longitudinal division can occur 
in either the nonflagellate (Fig. 1b) or the flagellate stage (Fig. le). 
As the organisms pass onward into the less favorable environment of 


Fig. 1—Herpetomonas: (a) non-flagellate or leishmaniform stage; (b) 
dividing non-flagellate; (c) elongating parasite; (d) flagellate stage; (e) divid- 
ing flagellate; (f) post-flagellate or encysted stage. 1500. 


the posterior end of the intestine of their host, their body cytoplasm 
concentrates, and the flagellum is withdrawn and largely dissolved. The 
now ovoid parasite secretes a coat which may be at first gelatinous but 
ultimately becomes varnish-like or “skin tight” and the postflagellate 
form is again produced. This resistant nonflagellate form (Fig. 1f) is 
particularly adapted for extracorporeal life and serves for the safe 
transference of the parasite from host to host. 

The above outline of the life history of a herpetomonad is valid for 
Herpetomonas jaculum, H. stratiomyiae, H. pediculi, H. culicis and H. 
ctenocephah with which we experimented. 

The life-history of a true Crithidia, such as C. gerridis, in its insect 
host has the same general outline as that of a Herpetomonas. But 
the flagellate stage differs from that of a Herpetomonas in that at the 
differentiation of a flagellum, this structure not only reaches the sur- 


FANTHAM-PORTER—EVOLUTION OF DISEASE 157 


face, but forces the ectoplasm before it, thus producing a small wavy 
undulating membrane that gradually fades into the free flagellum at 
the tapering anterior, flagellar end of the body of the organism (Fig. 
2b). 

The morphology of Herpetomonas jaculum, H. stratiomyiae, H. 
pediculi, H. culicis and H. ctenocephali in the vertebrate hosts into 
which they were introduced resembled that in the invertebrate hosts. 
The parasites have been introduced both as flagellates and as nonflagel- 
lates. In blood smears taken during the life of the host and in organ 
smears made at autopsy, usually both flagellate and nonflagellate forms 
were found. Parasites in various stages of multiplication were 


a b c 
Fig. 2.—Flagellate forms of (a) Herpetomonas (sometimes called Lepto- 
monas), (b) Crithidia and (c) Trypanosoma. X 2000. 


observed in the fresh condition and in stained preparations. Hence 
there is definite evidence that they had become true parasites of the 
vertebrates, had established themselves and had increased in numbers 
in them, and were not mere conservations of the forms introduced. 
The various Herpetomonas (Fig. 2a) and Crithidia (Fig. 2b) that 
we have used have retained the facies that they presented in the insect 
hosts. No transition to a trypanosome (Fig. 2c) was ever séen by us 
during the course of these experiments. The only variation presented 
by the parasites in the vertebrates from that in the invertebrates was 
that the maximum length of the insectan flagellate stage was not 
usually quite attained. The sizes of the parasites, however, were 


158 THE JOURNAL OF. PARASITOLOGY 


always well within the range of the limits of variation given for the 
insect parasites and were of good average size. Morphologically, they 
were replicas of the insect forms and could be unmistakably identified 
with them. The nonflagellate forms were about the size of or slightly 
greater than the Leishman-Donovan body in man, while the dimensions 
of the flagellate forms were much the same as those of Leishmania in 
cultures on the Novy-MacNeal-Nicolle medium, that is, about l5p 
to 20u in the long diameter of body. The slightly lesser dimensions of 
the parasites may be the results of transference and implanting of the 
organisms in new hosts, or perhaps the age of the host may influence 
the size of the parasite. It has been noticed that other parasites, for 
instance, certain Haemosporidia, introduced into unfamiliar vertebrates 
or into young hosts tend to produce new generations whose maximum 
dimensions are somewhat less than those of their progenitors. The 
same factors may apply in this induced herpetomoniasis. On the other 
hand, it is known that the nonflagellate parasite of Indian kala-azar 
maintained in dogs may increase in longest diameter from about 2.5y 
or 3.54 to 8u or Qu. Similar variations in size occur in the non- 
flagellate stages of closely allied herpetomonads in insects. 


THE SIGNIFICANCE OF CERTAIN NATURAL FLAGELLATES OF INSECTS IN 
THE EVOLUTION OF DISEASE 


The role of insects in the spread of disease among men and other 
animals has furnished some of the most important advances in knowl- 
edge made in recent times. Many parasitic protozoa are the descend- 
ants of free-living ancestors. The degrees of degradation from inde- 
pendent life to saprophytism and thence to parasitism are almost imper- 
ceptible but nevertheless exist. Neither are the grades of parasitism 
more well defined, and consequently a free-living organism that by acci- 
dent or chance has reached the alimentary tract of an insect may live 
there first as a saprophyte, feeding on the waste materials or the newly 
ingested food of the host. The minute quantities of nourishment lost 
by the host in this way become serious when cumulative, and the sapro- 
phytism then leads to parasitism of a somewhat low degree. When 
the living protoplasm of the host furnishes the nutriment required, the 
parasitism becomes obvious and the effects on the host are more or less 
marked. 

In the case of many intestinal flagellates of insects, the host has 
responded to the attacks of the parasites in such a way that a mutual 
toleration has become established between them. Under these circum- 
stances but little injury ensues to the host, and the flagellates con- 
cerned are considered as “natural” and practically harmless to the host. 
Further, they have often been considered as specific to the said hosts 
to which they are practically harmless. 


FANTHAM-PORTER—EVOLUTION OF DISEASE 159 


Should such flagellates reach a vertebrate host, two courses may 
result. In the first instance, the flagellates may merely perish. In 
the second case, should the introduced organism be sufficiently plastic, 
it may adapt itself to its new environment and be able to persist for 
a time. Should its powers of adaptation be marked, it will multiply, 
and the greater the rapidity of increase, the greater the danger to the 
host. In other words, environment and plasticity determine pathogen- 
icity. 

Certain of the flagellates show the transformation from almost 
harmlessness in the insect to pathogenicity in the vertebrate or newer 
host. The genus Herpetomonas affords a good example of the capacity 
for pathogenicity that may be latent in many organisms hitherto con- 
sidered harmless. 

Kala-azar, oriental sore and dermomucosal leishmaniasis are well- 
known tropical diseases due to members of the Herpetomonadidae that 
are known as Leishmania donovani, and L. infantum in the cases of 
kala-azar, and as L. tropica in the more local maladies of the skin. 
These organisms are, in all probability, herpetomonads of insects that 
have reached vertebrate hosts. It is known that the various species of 
Leishmania develop into typical herpetomonad flagellates in cultures, 
and for some time now these flagellate stages have been known in man. 
Thus, in 1911, Escomel saw flagellate forms of Leishmania tropica 
in man and published about them later. La Cava in 1912 described 
similar forms of the same parasite in Italy. Also in 1912, Splendore 
found elongating forms and a few flagellate parasites in dermomucosal 
leishmaniasis in Brazil, while Monge in 1914, when working on the 
same malady in Peru, found the herpetomonad stage of the parasite. 
Lately (September, 1915) Wenyon has found the flagellate stage of 
Leishmania donovani in a dog’subinoculated from other dogs, the strain 
being originally derived from a man who died of kala-azar contracted 
in Calcutta. Further, a new herpetomonad, Haemocystozoon bra- 
siliense, was found by Franchini in 1913 in a human subject. 

As a result of experimental work, such as that of Patton and of 
Wenyon, it has been shown that species of Leishmania can develop into 
herpetomonad flagellate stages within the intestines of certain insects, 
such as bedbugs and mosquitos (Stegomyia). The evidence that an 
ovoid Leishmania is the non-flagellate stage of a herpetomonas is 
proved, and the flagellate stage of Leishmania can exist in cultures, in 
insects and in man. Leishmania morphologically is a herpetomonad. 

Herpetomonads experimentally introduced into vertebrates by us 
have produced pathogenic effects recalling those of kala-azar. Both 
maladies present the same features—the insidious onset, the subsequent 
relatively rapid illness, the splenic and often hepatic enlargement, 
feverish attacks and emaciation. In the cases where chronic infections 


160 THE JOURNAL OF PARASITOLOGY 


were produced in our animals, the nonflagellate, leishmaniform stages 
of the parasites were more numerous, while in acute cases the flagellate 
forms were more obvious (see table). In the diseases due to Leish- 
mania spp. the flagellate forms in the vertebrate host are far less com- 
mon than the nonflagellate ones, but it is of distinct interest to note 
that Monge (1914) suggested that the presence of flagellate forms of 
L. tropica in man was an indication of increased virulence on the part 
of the parasite. Such an increased virulence certainly coincided with 
more marked development of flagellates in our animals. Though no 
general conclusion on the subject can yet be given, the hypothesis that 
the presence of flagellate Herpetomonas or Leishmania in the vertebrate 
host affords an index of virulence is supported by the experimental 
results that we have obtained. 

The part played by vertebrates proved capable of harboring herpet- 
omonads is one that demands the attention of all students of pre- 
ventive medicine and of sanitary reform. By experiment we have 
proved that flagellates belonging to the genera Herpetomonas and Cri- 
thidia have produced infections not only in mammalia like mice and 
dogs, but also in birds and in cold-blooded vertebrates such as members 
of the pisces, amphibia and reptilia. Further, these flagellates are cap- 
able of assuming resting, nonflagellate stages in these hosts. 

There is thus the possibility that various vertebrates—fish, amphibia, 
birds, reptilia, and mammals—may serve as reservoirs of the herpeto- 
moniases, including leishmaniases. The virus may be very attenuated 
and so escape detection, or only be revealed by the presence of flagellate 
forms in cultures. Recently (1914) Sergent, Lemaire and Senevet in 
Algeria have demonstrated the presence of a herpetomonad flagellate in 
the blood and organs of geckos obtained from areas in Algeria in 
which oriental sore due to L. tropica is present. Phlebotomus flies, 
which may harbor a natural herpetomonad, feed on the geckos and on 
man. Hence animals like geckos may possibly act as reservoirs of 
leishmaniasis. Chatton and Blanc (1914) have found possible leish- 
maniform bodies in the young red blood cells of geckos in Tunis. 
Bayon (1915) has found herpetomonad parasites in the cloaca of 
Chameleon pumilus at Robben Island, South Africa, and says that “it 
does not seem excluded that a chameleon can get infected through 
swallowing a fly containing Herpetomonidae in its gut.” He also 
found a herpetomonad in the gut of the fly, Scatophaga hottentota, 
in the same place. Lindsay (1914) stated that the parasite of dermo- 
mucosal leishmaniasis in Paraguay is believed by native sufferers to be 
conserved in rattlesnakes and to be spread by ticks or flies (Simulium) 
feeding on the reptiles and transferring the parasite to man. We 
have shown the possibility of such infection occurring by causing insec- 
tivorous vertebrates, such as viviparous lizards and grass snakes, to 


FANTHAM-PORTER—EVOLUTION OF DISEASE 161 


ingest insects infected with herpetomonads, wherewith the vertebrates 
became parasitised. Similarly, insectivorous birds have become para- 
sitised by ingesting insects containing herpetomonads. These infections 
could be accomplished in Nature and, in fact, such parasitism of a bird 
by herpetomonads and of mice by the same flagellates has been found 
(see below). Natural reservoirs of herpetomoniasis, consisting of 
vertebrates on which sanguivorous insects feed, should be sought for in 
areas where diseases such as kala-azar are present. 

Natural reservoirs of herpetomoniasis are already known. Man 
and his intimate domestic associate the dog, both may function as reser- 
voirs of what has been termed Mediterranean or infantile kala-azar. 
The parasite, Leishmania infantum, which is often considered to be a 
form of L. donovani, is thought to be transmitted from dog to dog by 
the dog flea and possibly also from dog to man. An infected child or 
an infected dog may, perhaps, serve as the reservoir of the virus. In 
this connection it is of some interest to recall that cattle which have 
become immune to piroplasmosis may yet harbor sufficient sparse piro- 
plasms in their blood to infect many ticks and so spread the malady. 
Analogy is somewhat dangerous, but in this case, it may be of service, 
since rare cases of “spontaneous cure” of infantile leishmaniasis are 
known and it is just possible that such may act as unsuspected reser- 
voirs of leishmaniasis. 

Vertebrates other than man can be infected naturally with herpet- 
omonads. In 1903, Dutton and Todd described herpetomonads from 
the blood of house mice in Senegambia. The original description was 
very definitely that of a Herpetomonas, though Todd has recently stated 
that he thinks the organism may have been a trypanosome. However, 
we have also found herpetomonads closely resembling those described 
by Dutton and Todd in mice in England. It is known that the common 
rat-fleas contain herpetomonads and it is suggested that these fleas 
were the probable source of infection. Mice as possible reservoirs of 
leishmaniasis cannot be disregarded. 

Again, a natural infection of birds has been described by Drs. 
Edmond and Etienne Sergent. In this case a pigeon was found to 
contain herpetomonads in its blood. The source of the flagellate is not 
known with certainty, but we advance the hypothesis that it was a 
latent herpetomoniasis contracted from herpetomonad-infected insects 
such as species of Lynchia that had fed on the bird. 

From a careful comparison of natural and induced herpetomoniasis 
in vertebrates and of leishmaniasis, as well as consideration of the 
morphology and life phenomena of the excitants in each case, the fol- 
lowing general statements can be made. Under suitable conditions, 
insect flagellates can be introduced into vertebrate hosts and can pro- 
duce infections therein. In some cases, as in some cold-blooded verte- 


162 THE JOURNAL OF PARASITOLOGY 


brates, little obvious ill effect results; in others, as in mammals and 
birds, disease is manifested and often ends in death. 

The organisms, such as herpetomonads, thus introduced, retain their 
powers of development on the same lines as when they were present 
in the insects. The morphological cycle is that of Herpetomonas. The 
various species of Leishmania are probably insect flagellates long since 
introduced into man and usually perpetuating the nonflagellate form, 
though capable of assuming the flagellate, herpetomonad facies in the 
internal organs of the vertebrate or in the invertebrate hosts. 

No insect flagellate can be considered to be quite innocuous to ver- 
tebrates until it has been put to the test. 

It must be remembered that leishmaniasis, which is a form of her- 
petomoniasis, is a flagellosis, as is also trypanosomiasis. The treatment 
of leishmaniasis by intravenous injection of tartar emetic—as advocated 
and practiced recently—is sound biologically, for drugs containing 
arsenic or antimony have proved efficacious in trypanosomiasis. 

It is necessary to consider not part, but the whole, of the life history 
of an organism and also the relationship of the parasite to the group to 
which it belongs. There is a line of evolution common to each group 
and in these cases, neither Herpetomonas (Leptomonas), Leishmania, 
Crithidia nor Trypanosoma (Fig. 2) should be considered as isolated 
units but as flagellates belonging to the Trypanosomidae. 


MODES OF INFECTION AND PREVENTIVE MEASURES AGAINST 
ARTHROPOD-BORNE HERPETOMONIASES 


The experiments on the introduction of various species of Herpet- 
omonas and Crithidia parasitic in insects into both warm and cold- 
blooded susceptible vertebrates has shown that these flagellates can 
produce an infection in the vertebrates when the latter are fed or 
inoculated with them. Within the host, the parasite is capable of 
assuming the leishmaniform or flagellate facies. The mode of infec- 
tion of the vertebrate in nature seems to be contaminative, either by its 
food, or through an already existing abrasion or puncture on the sur- 
face of its body. The feces of insects, if containing the resistant 
forms of the flagellate, are capable of producing infection by similar 
channels. We have also obtained evidence showing that postflagellate 
forms of the parasite are the best adapted to begin life in a new 
vertebrate host. 

Experiments on ourselves with fleas and lice, and with biting insects 
on rats, suggest that infection with Herpetomonas or Leishmania is not 
by inoculation with the protozoal parasites during the time when the 
insect is biting man or other vertebrate, but by the vertebrate eating the 
infected insect, or by infected insect feces passing through an abrasion, 
puncture or bite on the vertebrate skin. In this connection it is of 


FANTHAM-PORTER—EVOLUTION OF DISEASE 163 


interest to note that Laveran has quite recently succeeded in infecting 
a mouse with a culture of Leishmania tropica by way of the mouth. 

As we have already stated, in areas where leishmaniases are 
endemic, an examination should be made of all insects and other inverte- 
brates likely to come into contact with men or dogs or rats and mice, 
in order to ascertain if these invertebrates habor herpetomonads. Pre- 
ventive measures should be directed against such invertebrates, 
especially arthropods. Further, it is likely that certain vertebrates, 
such as reptiles and amphibia (especially such as are insectivorous), 
may serve as reservoirs for leishmaniases or, as they should preferably 
be termed, herpetomoniases. From such reservoirs the herpetomonads 
may reach man by the agency of ectoparasites or flies, especially such 
as are sanguivorous. 

That some of these suggestions are of practical application has 
been proved by the work of Dodds Price in the Assam tea gardens, fol- 
lowing on a suggestion from Rogers to the effect that action should 
be taken against suspected transmitters of kala-azar, even if complete 
inculpation of them had not been afforded. Dodds Price has reduced 
the mortality due to kala-azar enormously by segregating the infected, 
by moving coolie lines about three hundred yards from older, infected 
ones and by having new coolie lines placed on clean sites. Young 
(1914) has applied successful segregation measures to an indigenous 
population in certain villages in Assam. These measures check the 
prevalence of sanguivorous insects that infest man and his dwellings, 
and reduce the danger of possible infection by way of contaminated 
food or drink. It may be expected that the application of similar 
measures in other areas where kala-azar is endemic may also be equally 
efficacious. 

SUMMARY 


1. Herpetomoniasis can be induced in various warm and cold- 
blooded vertebrates when the latter are inoculated or fed with herpet- 
omonads occurring in the digestive tracts of various insects. The 
infection produced and the protozoal parasites found in the vertebrates 
resemble those of human and canine leishmaniases. 

2. An infection can also be induced in certain vertebrates when they 
are fed or inoculated with Crithidia gerridis, and both flagellate and 
nonflagellate stages occur therein, but no transition to a trypanosome 
was found. 

3. The following Flagellata have been proved pathogenic to warm- 
blooded vertebrates when the latter have been fed, or inoculated sub- 
cutaneously or intraperitoneally with them—Herpetomonas jaculum, 
H. stratiomyiae, H. pediculi, H. ctenocephali, H. culicis and Crithidia 
gerridis. The hosts used were mice of various ages, dogs, canaries, 
sparrows and martins. 


164 THE JOURNAL OF PARASITOLOGY 


4. Herpetomonas jaculum and Crithidia gerridis have also been suc- 
cessfully fed or inoculated into cold-blooded hosts, namely, fishes 
(Gasterosteus aculeatus), frogs, toads, lizards (Lacerta vivipara) and 
grass-snakes (Tropidonotus natrix). 

5. The disease induced may run an acute or a chronic course. In 
the acute cases among our vertebrates the flagellate form of the para- 
site was the more obvious at death. In chronic cases, non-flagellate 
forms of the parasite were more numerous. 


6. Natural herpetomoniasis of a pigeon has been recorded by Drs. 
Edm. and Et. Sergent in Algeria. This affords a parallel case with the 
natural and induced herpetomoniasis of mice as recorded by us. 


7. The flagellate stage of Leishmania donovani in vertebrates is 
now known, and that of L. tropica in man has been known for some 
time. The links completing the evidence that a Leishmania is morpho- 
logically a Herpetomonas are thus complete. We believe that leish- 
maniases are invertebrate-borne herpetomoniases, and that these mala- 
dies have been evolved from flagellates of invertebrates (especially 
herpetomonads of insects), which have been able to adapt themselves 
to life in vertebrates. 


8. In areas where leishmaniases are endemic an examination should 
be made of all insects and other invertebrates likely to come into con- 
tact with men or dogs or domestic vermin like rats and mice, in order 
to ascertain if these invertebrates harbor herpetomonads. Preventive 
measures should be directed against such invertebrates, especially 
arthropods. Further, it is likely that members of all classes of verte- 
brates, and especially those members that are insectivorous, may serve 
as reservoirs for leishmaniases, or as they should preferably be termed, 
herpetomoniases. The virus may exist in such reservoirs in a very 
attenuated condition and so be difficult of detection. From these 
sources the herpetomonads may reach man by the agency of ectopara- 
sites or flies, especially such as are sanguivorous. 


ADDENDUM 


As this paper—the writing of which has been greatly delayed by 
war work-—was on the point of being despatched, our attention was 
drawn to an article on The Insect Vector of Uta by C. H. T. Townsend 
in the December number of the Journal of Parasitology, just received 
in England. The concluding paragraph of the text and more particu- 
larly of the summary of Townsend’s paper were read by us with very 
great interest, as they confirm our conclusions regarding leishmaniases 
being arthropod-borne herpetomoniases. This conclusion of ours has 
met with considerable opposition at the hands of Wenyon, much to our 


FANTHAM-PORTER—EVOLUTION OF DISEASE 165 


surprise, and in spite of the fact that the experiments of Laveran and 
Franchini, as well as the mttch more extended series of our own, admit 
of no other conclusion to our mind. 

The following conclusions of ours may be compared with those of 
Townsend (December, 1915). Thus, in November, 1914, we stated 
that, “It may be expected that the various leishmaniases, occurring in 
different parts of the world, will prove to be insect-borne herpeto- 
moniases.” Again, in May, 1915, we wrote that: “As we have previ- 
ously stated, we believe that leishmaniases are arthropod-borne herpet- 
omoniases, and that these maladies have been evolved from flagellates 
of invertebrates (especially herpetonomads of insects), which have 
been able to adapt themselves to life in vertebrates.” Further, one of 
us in June, 1915, wrote that: “It is inferred that the various leish- 
maniases are due to a herpetomonad of invertebrates which, under dif- 
ferent conditions of environment, produces pathogenic effects in very 
varying degrees in different vertebrates, from zero, as in the mice 
described by Dutton and Todd in 1903, to high mortality as in Indian 
kala-azar, and probably zero again in cold-blooded hosts. It is also a 
flagellate which can probably live in invertebrates not already recorded 
as being infected. A human reservoir of leishmaniasis may occur in 
some places, while warm and cold-blooded vertebrates may also func- 
tion-as the same.” 

REFERENCES CITED 

Bayon, H. 1915. Herpetomonidae found in Scatophaga hottentota and 
Chameleon pumilus. Trans. Roy. Soc. S. Africa, 5: 61-63. 

Chatton, E., and Blanc, G. 1914. Existence de corps leishmaniformes dans 
les hématoblastes d’un Gecko barbaresque, Tarentola mauritanica. C. R. Soc. 
Biol., 77 : 430-433. 

Dutton, J. E., and Todd, J. L. 1903. First Report of the Trypanosomiasis 
Expedition to the Gambia (1902). Part of Sec. VII. Flagellata in the Blood 
of a Mouse. Liverpool Sch. Trop. Med., Memoir, 11: 56-57. 

Escomel, E. 1914. Leishmania Flagelada en el Peru. La Cronica Medica 
(Lima), 31: 224-227. 

Fantham, H. B. 1912. Herpetomonas pediculi, nov. spec., parasitic in the 
alimentary tract of Pediculus vestimenti, the human body louse. Proc. Soc. 
Lond., B. 83: 212-227; 1 pl. 1915. Insect Flagellates and the Evolution of Dis- 
ease, with Remarks on the Importance of Comparative Methods in the Study 
of Protozoology. Annals Trop. Med. and Parasitol., 9: 335-348. 

Fantham, H. B., and Porter, A. 1914. Some Insect Flagellates Introduced 
Into Vertebrates. Proc. Cambr. Philos. Soc., 18: 39-50; 1 pl. 1915. Further 
Experimental Researches on Insect Flagellates Introduced Into Vertebrates, 
Proc. Cambridge Philosoph. Soc., 18: 137-148. 1915a. On the Natural Occur- 
rence of Herpetomonads (Leptomonads) in Mice. Parasitology, 8: 128-132. 
1916. Some Experimental Researches on Induced Herpetomoniasis in Birds. 
Annals Trop. Med. and Parasitol., 9: 543-558; 1 pl. 

Franchini, G. 1913. Un nouveau protozoaire parasite de l’>homme provenant 
du Brésil. Bull. Soc. Path. Exot., 6: 156-158. 

Laveran, A., and Franchini, G. 1914. Infections de Mammiféres par des 
flagellés d’invertébrés. Bull. Soc. Path. Exot., 7: 605-612; 4 figs. 


166 THE JOURNAL OF PARASITOLOGY 


Monge, C. 1914. La Leishmaniasis en el Peru. Espundia, Uta, Juccuya, 
Qcepo, Tiacc-arafia. La Cronica Medica (Lima), 31: 231, 251, 288, 385. 

Porter, A. 1911. The Structure and Life History of Crithidia pulicis, n. sp., 
Parasitic in the Alimentary Tract of the Human Flea, Pulex irritans. Para- 
sitology, 4: 237-254; 1 pl. 

Price, J. Dodds, and Rogers, L. 1914. The Uniform Success of Segregation 
Measures in Eradicating Kala-azar from Assam Tea Gardens. Its Bearing on 
the Probable Mode of Infection. Brit. Med. Jour., Feb. 7, pp. 285-289. 

Sergent, Edm., and Et. 1907. Etudes sur les Hématozoaires d’Oiseaux. 
Ann. Inst. Pasteur, 21: 251-280; 2 pls. (For natural Herpetomonad in pigeon, 
see footnote, p. 270 and Plate VII). 

Wenyon, C. M. 1915. Flagellate Form of Leishmania donovani in the Tis- 
sues of an Experimentally Infected Dog. Jour. Trop. Med. and Hyg., 18: 218-219. 


A REVISION OF THE GENUS ARHYTHMORHYNCHUS 


WITH DESCRIPTIONS OF TWO NEW SPECIES FROM NORTH 
AMERICAN BIRDS * 


H. J. VAN CLEAVE 


INTRODUCTION 


When Lithe created the genus Arhythmorhynchus (1911: 47) he 
assigned to it but one species A. frassoni (Mol.). The following year 
in publishing the results obtained from a study of four immature speci- 
mens of Echinorhynchus invaginabilis von Linstow Lithe (1912:283) 
ascribed that species to the genus Arhythmorhynchus and in the same 
article accepted two American species, Echinorhynchus wuncinatus 
Kaiser and E. trichocephalus R. Leuckart, as agreeing with his defini- 
tion of the genus Arhythmorhynchus. Of these four species belonging 
to this genus but one is well known, namely, A. frassoni (Mol.). For 
the two American species not even the host is known, and while Kaiser 
(1893) has given minute details regarding the hooks of these two 
species, data concerning the embryos and many other points which are 
essential for a complete specific diagnosis are entirely wanting. Con- 
sequently it seems that concerning some of the points in the definition 
of the genus data are available for a single species only. It is not 
surprising that a generic diagnosis based upon the study of a very small 
number of species might later require emendation to permit including 
within the same genus species of obviously close relationship. Espe- 
cially is this true in groups of parasites, such as the Acanthocephala, in 
which the organization of the body has been reduced to its simplest 
terms through perfect adaptation to the parasitic existence; for this 
same reduction eliminates large groups of organs and structures which 
in nonparasitic forms afford additional characteristics of diagnostic 
value. 

‘Recently the writer (Van Cleave, 1913) found it advisable to emend 
the definition of the genus Neoechinorhynchus to permit including 
within it five species which were unknown to the founder of the genus. 
Similarly now after a study of new materials including two new species 
closely related to Arhythmorhynchus frassoni (Mol.) and A. invagina- 
bilis (von Linst.) the writer has found it imperative to modify. Lithe’s 
original description of the genus Arhythmorhynchus (Liihe, 1911: 47) 
to prevent exclusion from this genus of forms which under a natural 


* Contributions from the Zoological Laboratory of the University of Illinois, 
No. 66. 


168 THE JOURNAL OF PARASITOLOGY 


system of classification could not be granted independent generic rank. 
The materials upon which the present study has been made were col- 
lected by Mr. Albert Hassall and deposited in the Collections of the 
U. S. Bureau of Animal Industry. Both species are represented in the 
collection by numerous fully mature individuals so that a complete 
study of the specific characteristics has been possible in both species. 
These new forms were found to deviate from Luhe’s description of the 
genus Arhythmorhynchus in the following particulars: (1) the shape 
of the body; (2) the location of the testes with reference to the two 
regions of the body proper described by Lthe; (3) the shape of the 
membranes surrounding the hard-shelled embryos in the body cavity 
of the female. In the estimation of the writer these are placed in the 
order of their relative significance, the least significant first. 

As Lithe has pointed out, the anterior region of the body in mem- 
bers of the genus Arhythmorhynchus, which in some species is an 
inflated oval region standing out in contrast to the smaller cylindrical 
posterior region (Figs. 1 and 4) contains relatively large numbers of 
subcuticular nuclei while the posterior region is devoid of subcuticular 
nuclei. Simple body shape has been so long recognized as a variable 
quantity by those working with Acanthocephala that little emphasis 
may justly be given it alone. However, when body shape has as an 
accompanying feature distinctive structural characteristics emphasis 
may be placed upon the structure as of diagnostic value though broad 
range of variation may occur in the gross outer form in which the 
structure finds expression. Therefore, in defining the genus Arhythmo- 
rhynchus, emphasis should be placed upon the difference in structure 
between anterior and posterior regions of the body rather than upon the 
difference in shape of these two regions, for the structure is constant in 
all species which have been examined though the body form is widely 
variable. For the males of this genus Lthe has specified that the testes 
occur in the anterior swollen region of the body (Fig. 1). Suspended 
as they are in the genital ligament running backward through the body 
cavity from the base of the proboscis sheath and with no intimate rela- 
tionship to the body wall little of generic value may be placed upon the 
exact location of these organs in the body cavity. In one species at 
least (Fig. 4) the testes do not lie in the swollen region of the body as 
indicated in Luhe’s diagnosis of the genus, but are located distinctly 
posterior to that region. The third point of difference, the shape of the 
embryonic membranes, presents the most radical point of divergence in 
the species under consideration from the original description of the 
genus. Luhe throughout his classification of the Acanthocephala has 
emphasized the importance of shape of the embryos and structure of 
their coverings as of marked diagnostic value. For Arhythmorhynchus 
he has specified in his characterization of that genus the presence of 


VAN CLEAVE—REVISION OF GENUS ARHYTHMORHYNCHUS 169 


three fully concentric membranes surrounding the embryo within the 
body cavity of mature females. His observations upon the embryos 
of A. frassoni, the only species of the genus for which sexually mature 
individuals were at that time known, corroborated the earlier record of 
de Marval (1904, Fig. 55) for the embryos of the same species. How- 
ever in A. brevis and A. pumilirostris the writer has found two species 
which, though agreeing with Liihe’s definition of the genus in all other 
essential characteristics, present a marked contrast in the structure of 
the embryos. In each of these species numerous fully mature females 
have been examined with the unvarying result of disclosing hard-shelled 
embryos in which the middle membrane has an outpocketing at each 
pole (Figs. 10 and 12). 

In view of the foregoing details, wherein the two newly described 
species fail to agree with the original definition of the genus, two pos- 
sibilities present themselves: either (1) a new genus should be estab- 
lished for these two species; or (2) the definition of the genus 
Arhythmorhynchus should be modified so as to include these forms. 
To the writer it seems unwise to create a new genus for forms which 
differ from an existing genus by but a single point of essential distinc- 
tion: namely the shape of the membranes surrounding the embryos. 
Especially does this seem uncalled for in the case under consideration 
in which up to the present time, embryos were known for but a single 
species. Therefore it appears expedient to recast the definition of the 
genus Arhythmorhynchus. 


REVISED DIAGNOSIS OF THE GENUS ARHYTHMORHYNCHUS 


Acanthocephala with a spindle-shaped proboscis upon which the 
hooks are arranged not in radial but in bilateral symmetry since those 
on the dorsal and ventral surfaces of the same individual differ, though 
in varying degrees in different species. Anterior region of body sharply 
differentiated from posterior region in structure of body wall, espe- 
cially in the presence of nuclei in the subcuticula of anterior region 
only. Portion of the anterior region of body proper spined. Spines 
entirely wanting on neck and on posterior region of body proper. 
Proboscis sheath a double-walled muscular sac inserted at the base of 
proboscis. Central nervous system near center of proboscis sheath. 
Cement glands very long, slender. Embryos in body cavity of female 
elongated oval with three membranes, all concentric or the middle one 
with an outpocketing at each pole. Sexually mature in the intestine 
of birds. 

ARHYTHMORHYNCHUS BREVIS NOV. SPEC. 

Body in both sexes with distinct oval enlargement comprising about 
anterior half. Posterior end distinctly smaller, elongated, cylindrical. 
Females 6 to 12 mm. long; maximum thickness 3 mm.; diameter of 


170 THE JOURNAL OF PARASITOLOGY 


posterior region about 1 mm. Males, 5 to 6 mm. long; maximum thick- 
ness, 1 to 1.5 mm.; diameter of posterior region, 0.5 to 0.75 mm. 
Neck naked, retractable, tapering toward proboscis, not sharply set off 
from body, 0.35 to 0.55 mm. long. Body for short distance just back 
of neck irregularly set with small number of spines 0.012 mm. long. 
Proboscis elongated with conspicuous expansion near center, 0.665 mm. 
long, 0.230 mm. in diameter at base, 0.190 mm. at tip, 0.340 mm. at 
center. Proboscis armed with eighteen longitudinal rows of hooks, 
usually fifteen in a row. Basal hooks nearly straight, slender, 0.047 
mm. long. Heaviest hooks near middle of proboscis 0.041 to 0.047 mm. 
long, on ventral surface slightly larger than on dorsal. Hooks at tip 
slender, recurved, 0.047 mm. long. Cement glands long, narrow. Testes 
oval, slightly overlapping one another, in swollen part of body. 
Embryos 0.076 to 0.100 mm. by 0.024 to 0.030 mm. Middle of three 
shells of embryos heavy, with a rounded swelling at each pole. Host 
Botaurus lentigenosus (Montag.), intestine. Type locality Baltimore, 
Md., U.S. A.; Cotypes in collection Bureau of Animal Industry, Wash- 
ington, D. C., Catalog No. 6302; and in the Helminthological Collection 
of the Department of Zoology, University of Illinois, Urbana, Catalog 
No. 16, 165. 

The structure of the body wall in the genus Arhythmorhynchus 
presents numerous anomalies when compared with conditions found in 
other genera of Acanthocephala. The writer has made a study of some 
of these points, especially in the species A. brevis, the results of which 
follow. Lithe (1911: 47) has called attention to the peculiar distribution 
of the subcuticular nuclei in this genus and incidentally in a vague man- 
ner has referred to other differences between the anterior and posterior 
regions of the body. Figure 11 shows the shape, structure, and loca- 
tion of the subcuticuiar nuclei in a longitudinal section through the 
anterior region of a specimen of A. brevis. The entire subcuticula is 
a peculiar structure, presenting an appearance unparalleled in any other 
genus of Acanthocephala. In the anterior region of the body there may 
be easily recognized beneath the cuticula (c) a region in which small 
fibrillae run both longitudinally and radially (scr). An intermediate 
more heavily granular zone (sc2) separates this region from the region 
of radial fibers (sc?) in which the subcuticular nuclei (sv) are con- 
tained. This last region, which Kaiser (1913, Plate 1, Fig. 1) in Gigan- 
torhynchus hirudinaceus called the hypoderm, is bounded on its inner 
surface by a layer of circular muscular threads (cmt). 

The longitudinal muscular layer in the anterior part of the body 
shows some most striking deviations from conditions usually found in 
the body musculature of Acanthocephala. The presence of large nuclei 
(mn) in, and of numerous finger-like fiber-bundles (mf) imbedded in 
an undifferentiated cytoplasmic envelop (uc) suggest a resemblance to 


VAN CLEAVE—REVISION OF GENUS ARHYTHMORHYNCHUS 171 


the nematode musculature. But this can be scarcely more than a sug- 
gestion since the orientation of the fibers is the opposite of that charac- 
teristic of the nematodes. Figure 11, a longitudinal section of A. brevis, 
shows these fiber-bundles in a position comparable to the view obtained 
in a cross section of a nematode. Though this agreement in: fundamen- 
tal structure of the muscle cells may indicate a relationship between 
the Acanthocephala and the Nematoda, yet the confusion in the 
arrangement of the fibers prevents ascribing to the argument any great 
phylogenetic importance. 

Lithe in his characterization of the genus Arhythmorhynchus com- 
mented upon the slight development of the lacunar system of the sub- 
cuticula. In Figure 7, the writer has shown a portion of a tangential sec- 
tion through the subcuticula of A. brevis. A longitudinal canal (Jc) 
is shown in its characteristic relationship with a circular canal (cc). 
In this species, at least, the canal system is well developed, though the 
extent and complexity of the subcuticular layer tend to make it incon- 
spicuous. 

In the posterior region of the body the body-wall presents its broad- 
est departure in A. brevis from the conditions usually found in other 
genera. Here, as- has been stated before, there are no subcuticular 
nuclei. The regions of the subcuticula (Fig. 8) agree in arrangement 
and general structure with those previously described for the same 
layer in the anterior part of the body. However, between the double 
row of circular muscle threads (cmt) and the body cavity is interposed 
a series of structures which are evidently modified continuations of 
the muscular system described for the anterior region of the body. 
In a longitudinal section, or in an optical section of a well prepared 
whole mount, this modified part has the appearance of a series of 
triangular elevations (tr) with the base of each triangle directed 
toward the layer of circular muscle threads. From the apex of each 
of these triangles is given off a fine membrane (m) which runs inward 
toward the longitudinal muscle sheath. Each of the triangular eleva- 
tions is pierced by a canal (ca) about 0.025 mm. in diameter. These 
triangular ridges occupy only about one fourth the region between the 
circular muscular threads and the muscle sheath lining the body cavity. 
Most of the intervening space is open cavity intercepted at irregular 
intervals by very thin membranes (ms) of another series which do not 
take their origin or have their insertion in the triangular ridges. The 
open spaces between the membranes are in communication with the 
central body cavity as is especially shown by the presence within the 
chambers of large numbers of eggs and embryos (e) in various stages 
of formation. 

Some of the muscles within the body cavity show a peculiar stria- 
tion. Figure 6 represents a single fiber of one of the retractor muscles 


172 THE JOURNAL OF PARASITOLOGY 


greatly magnified. Regions of dark striations (st) alternate with bands 
of nonstriated structure, while the nucleus (7) is in a mass of undif- 
ferentiated cytoplasm at one side of the fiber. 


ARHYTHMORHYNCHUS PUMILIROSTRIS NOV. SPEC. 


Body of males and immature females with slight enlargement com- 
prising about anterior fifth. Gravid females with posterior region of 
body enlarged, cylindrical, with irregularly distributed swellings. 
Females up to 30 mm. long. Maximum diameter fully gravid female, 
slightly posterior to middle of body, 1.5 mm.; diameter anterior region 
0.9mm. Neck naked, retractile, tapering toward proboscis; in size not 
sharply set off from body. Body for short distance behind neck set with 
small spines, 0.012 to 0.020 mm. long. Proboscis elongated, with con- 
spicuous swelling near center; length 0.450 mm.; maximum breadth 
0.180 mm.; breadth at tip 0.095 mm., at base 0.114 mm. Proboscis 
armed with sixteen longitudinal rows of hooks with fourteen or fifteen 
hooks in a row. Basal hooks nearly straight, thorn like, usually 0.035 
mm. long. Heaviest hooks on ventral surface near middle of proboscis 
0.030 mm. long. Hooks at tip slender, recurved, 0.030 to 0.035 mm. 
long. Cement glands in male extremely attenuated. Testes contigu- 
ous in region behind anterior swelling of body. Embryos 0.065 to 0.089 
mm. long; 0.018 mm. wide; with three membranes, the middle one 
with an outpocketing at each pole. 

Host Botaurus lentigenosus (Montag.), intestine. Type locality 
Washington, D. C. Cotypes in collection of Bureau of Animal Indus- 
try, Washington, D. C., Catalog No. 2076; and in the Helminthological 
Collection of the Department of Zoology, University of Illinois, Catalog 
No. 16, 166. | 

In its microscopic anatomy this species closely resembles that given 
for the preceding species. Figure 13, an optical section of A. pumili- 
rostris, indicates the general distribution of the two types of subcuticu- 
lar structure discussed under the morphology of A. brevis, while Figure 
12 shows a single hard shelled embryo. 


INTERRELATIONSHIPS OF THE SPECIES 


Upon the basis of the characteristics of the proboscis hooks alone 
there is an indication of a natural division of this genus into two sub- 
groups which make comparisons between species fairly certain even 
though the essential diagnostic facts for some species are not all known. 
One group consists of those species whose members possess a few 
extremely large hooks at the middle of the ventral surface of the pro- 
boscis; A. frassoni and A. trichocephalus fall within this group. In 
the second group the midventral hooks are but slightly larger than the 


VAN CLEAVE—REVISION OF GENUS ARHYTHMORHYNCHUS 173 


midlateral and middorsal hooks; to this belong A. imvaginabilis, A. 
brevis, A. uncinatus, and A. pumilirostris. A. brevis and A. pumili- 
rostris may be separated from A. invaginabilis upon the basis of the 
number of longitudinal rows of hooks upon the proboscis. For the last 
named species Lithe (1912:287) found twenty-two to twenty-four 
longitudinal rows of hooks. Eighteen are found in A. brevis and 
sixteen in A. pumilirostris. The separation of 4. uncinatus is most 
sharply shown in a comparison of the size of the hooks. Kaiser 
(1893: 15) found hooks upon the proboscis of A. uncinatus ranging 
from 0.056 to 0.120 mm. long while in A. brevis the writer has found 
the range in size of hooks to be from 0.030 to 0.047 mm., and in A. 
pumilirostris the longest hooks are 0.035 mm. long. A. brevis and 
A. pumilirostris are most readily separable one from the other by the 
fact that the former has the larger proboscis with eighteen longitudinal 
rows of hooks, while the latter has but sixteen longitudinal rows of 
hooks upon a much smaller proboscis. 

At the end of his work on the Acanthocephala of the fresh waters 
of Germany, Lithe (1911:53) has considered a number of species 
which were insufficiently known to permit of classification in his system 
with certainty. Among these is a species E. striatus Gze. for which 
he has mentioned an apparent relationship with the genus Corynosoma 
through the shape of the embryos. Since this is the sole point where 
the present writer found the two species 4. brevis and A. pumuilirostris 
to differ from Lithe’s description of the genus Arhythmorhynchus and 
since the figures and description of E. striatus agree also with that 
genus the writer can see no objection to including the species striatus 
within the genus Arhythmorhynchus as emended in the present paper. 


KEY TO THE SPECIES OF ARHYTHMORHYNCHUS REPORTED FROM 
NORTH AMERICA 


—" 


(2) Hooks on mid-ventral surface of proboscis conspicuously larger 
Hiesieany Orners 24S. 2 A. trichocephalus (R. Leuckart) 
(1) Hooks on ventral surface of proboscis not conspicuously larger 
prissaetae eee GSUIETACES. . Ve tet cine et ee eee et wenes 3 
(4) Longest hooks more than 0.100 mm....A. uncinatus (Kaiser ) 
(3) Longest hooks not more than 0.050 mm......-.-.++++++++- 5 
(6) Proboscis with sixteen longitudinal rows of hooks; embryos 
0.065 to 0.089 mm. long and 0.018 mm.wide.....-.-+-++++++++: 
Meee Onc hoe Ee ee Oe A. pumilirostris Van Cleave 
6 (5) Proboscis with eighteen longitudinal rows of hooks, embryos 
0.076 to 0.100 mm. long, and 0.024 to 0.030 mm. wide........ 
ee Ee ee ee ee a eee A. brevis Van Cleave 


bo 


wm & W 


174 THE JOURNAL OF PARASITOLOGY 


SUMMARY 


Two new species of Acanthocephala from the intestine of Botarurus 
lentigenosus show close relationship to Arhythmorhynchus frassoni. 
They fail to agree with Luthe’s definition of the genus Arhythmo- 
rhynchus in: (1) shape of the body; (2) location of the testes; (3) 
shape of the membranes surrounding the hard-shelled embryos. The 
original characterization of the genus is emended to include these forms 
which possess every other essential characteristic of the genus. 


LITERATURE CITED 


Kaiser, J. E. 1893. Die Acanthocephalen und ihre Entwicklung. Biblioth. 
Zool., 7: 1-136. 1913. Die Acanthocephalen und ihre Entwicklung. Leipzig, 
66 pp. 

Linstow, O. von. 1902. Beobachtungen an neuen und bekanten Helminthen. 
Arch. mikr. Anat., 60: 217-232. 

Lihe, M. 1911. Die Sitisswasserfauna Deutschlands, Hefte 16, Acantho- 
cephalen. Jena; 116 pp. 1912. Zur Kenntnis der Acanthocephalen. Zool. 
Jahrb., Suppl. 15; 1: 271-306. 

Marval, L. de. 1905. Monographie des acanthocéphales d’oiseaux. Rev. 
suisse de zool., 13: 195-387. 

Van Cleave, H. J. 1913. The Genus Neorhynchus in North America. Zool. 
Anz., 43: 177-190. 


EXPLANATION “OF PLATES 


All figures drawn from permanent, stained, balsam mounts with the aid of 
a camera lucida. 
PLATE 1 


Figs. 1 to 3—Arhythmorhynchus brevis nov. spec. 

Fig. 1—Immature male, entire. 

Fig. 2—Profile, dorsal surface, proboscis of mature male. 

-Fig. 3.—Profile, ventral surface, same proboscis as in Figure 2. 

Figs. 4 and 5.—dArhythmorhynchus pumilirostris nov. spec. 

Fig. 4—Male, entire. 

Fig. 5.—Profile, anterior end of body of same individual as shown in Figure 
4. Proboscis hooks same magnification as Figures 2 and 3 of A. brevis. 

Figs. 6 and 7.—A. brevis. 

Fig. 6—Muscle fiber from one of the retractor muscles. 

Fig. 7.—Portion of tangential section through cuticula and subcuticula show- 
ing relations of longitudinal (/c) and circular (cc) canals. 


PLATE 2 

Figs. 8 to 11.—A. brevis. 

Fig. 8—Portion of body wall in posterior region. Sagittal section. For 
details see text. 

Fig. 9—Spines from anterior part of body wall. 

Fig. 10—Embryos from gravid female. 

Fig. 11—Portion of body in anterior region. Sagittal section. For details 
SEG ext, 

Figs. 12 to 14—A. pumilirostris. 

Fig. 12—Embryos from gravid female. 

Fig. 13—Anterior end of body, optical section, showing relative differentia- 
tion and distribution of subcuticula. 

Fig. 14—Spines from anterior part of body. 


PLATE 1 


Be 


\ 


PLATE 2 


0.05 ™% 


SOME NOTES ON THE ENCYSTED LARVA OF THE 
LUNG DISTOME 


SADAO YOSHIDA 
Pathological Department, Osaka Medical Academy 


In a former article (1916) I reported on (1) the discovery of the 
intermediate hosts (crabs) of the lung distome in Japan; (2) species 
of the intermediate hosts in various districts of our country; (3) the 
frequency of occurrence of the encysted larvae (cysts) in various 
crabs; (4) morphology of the encysted larva; (5) the animals, experi- 
mentally fed with the cysts, etc. The report to be given in the follow- 
ing pages is a part of the results obtained by subsequent study on the 
cysts of the lung distome in crabs, especially Eriocheir japonicus (de 
Haan). 


DISTRIBUTION AND MIGRATION OF CYSTS IN THE BODY OF AN 
INTERMEDIATE HOST 


The encysted larvae are found in various parts of E. japonicus, 
namely, muscles, hypodermis, gills, liver and other organs; of these, 
muscles and gills are most abundantly infected. The absolute number 
of cysts is greater in the muscles than in the gills, but the relative num- 
ber is inverse, because the volume of the muscles is much larger than 
that of the gills. The cysts in the gills are found only along a limited 
portion of blood vessels running longitudinally on the median line of 
their upper surfaces. In the muscles the cysts are found most fre- 
quently and most abundantly in the base of each appendage. Numer- 
ous cysts are often found in the muscles near the basipodite of each 
appendage, even in the cases in which a few or none of cysts are found 
in other parts of the musculature. 

From the abundance of the cysts in gills and near their attachment, 
the basipodite of each leg, and from the system of blood circulation in 
the crab, I am inclined to believe that the encysted larvae have a ten- 
dency to migrate toward the gills from all parts of body by means of 
the blood circulation. It was experimentally proved that the cyst has 
the ability to migrate through the various tissues of the crab, although 
the rate of migration is very slow. On the other hand, the circulatory 
system of the crab is open, as the distal ends of the arteries open into 
the tissues of the body and thus all tissues are bathed in the blood. 
The venous system begins not with capillaries, as in a closed system, 
but with lacunae, lying irregularly among the tissues. The lacunar 
spaces in the tissues communicate with one another at first, and gradu- 


176 THE JOURNAL OF PARASITOLOGY 


ally form a canal system after union of several lacunae from different 
parts; ultimately these grow into the venous vessels which run toward 
the gills to purify the blood. The blood current among the tissues and 
in the vessels of the venous system surely facilitates the migration of 
the cysts toward the gills. If this supposition is right, it explains 
clearly why the cysts are found abundantly in the small blood vessels 
in the gills and in the muscle near the base of each appendage. 

Thus the venous vessel is the most convenient course by which the 
cysts migrate toward the gills. For what purpose do the cysts migrate 
to the gills? Is there any necessity for the cysts to migrate to the gills? 
It is most favorable, I believe, for the cysts to migrate to the gills in 
order to facilitate further development of the encysted larvae by getting 
into the final host. On the whole, there are two ways by which the 
cysts may be taken up by the final hosts—human beings or other 
animals as dog, cat, etc.—namely: (1) their consumption as a food in 
an uncooked crab; (2) being taken with food and drink infected with 
cysts liberated into water from the intermediate host. In the second 
method of infection it is necessary for the cysts to escape into the 
water from the infected crab. The gills are the most convenient point 
at which the cysts can escape easily into the water, because the organ 
is always being laved by water and the blood vessel containing the cysts 
is separated from the outside water only by the very thin membranous 
wall. Thus it is reasonable to think that the cysts in various parts of 
the intermediate host migrate through the tissues carried by the blood 
current in the venous vessels toward the gills from which they may be 
discharged into the water. 

It is questionable in my mind whether the cysts in a crab (E. 
japonicus) escape into the water naturally and actively to secure an 
opportunity of being taken up by the final host. In Corea, R. Moriyasu, 
E. Arima, and M. Tanaka, proved experimentally the natural and active 
escape of cysts in the case of E. japonicus. In Formosa, K. Nakagawa 
obtained the same results as Moriyasu in the case of P. obtusipes 
(Stimpson). In Japan proper, R. Ando also proved experimentally 
that the result is quite the same in the case of P. dehaanti (White). All 
these writers made their experiments by approximately similar methods, 
namely, putting ten to thirty specimens of a crab which seemed to be 
infected with the cysts into cylindrical glass vessels with a little water. 
Renewing the water once or twice a day, they searched for cysts. In 
these examinations they all found the cysts more or less numerous, and 
hence concluded that the cysts escaped naturally and actively from the 
body of crab. In the case of P. dehaanii and P. obtusipes, it is possible 
that the cysts in the crab may escape into the water naturally and 
actively, because in these intermediate hosts the cysts are often found 


* a 


YOSHIDA—ENCYSTED LARVA OF LUNG DISTOME 177 


attached to the outer surface of gills, as I reported in the former paper 
(1916). ' 

I have also made some experiments to prove the natural and active 
discharge of cysts from the crab. There were two sets of experiments: 
1. I prepared a glass aquarium 75 cm. long, 27 cm. wide and 29 cm. 
deep, provided with three small exits on the bottom, the upper side 
being open and covered by metal gauze when necessary. Twenty 
crabs or more of moderate size were put into the aquarium, and 
water was permitted to flow in by a pipe and out through three exits 
which were closed by two or three sheets of gauze in order to prevent 
the escape of the cysts. I examined the sediment on the bottom occa- 
sionally for cysts and have often found them among materials there. 
2. I put one or two specimens of crab into a cylindrical glass vessel 18 
cm. in diameter and 15 cm. deep, pouring in water to a depth of 3 cm. 
or more. I prepared five such vessels, and renewed the water once or 
twice a day. This set of experiments was continued three months, 
having been started October 4 last year. In this long-continued experi- 
ment, only one cyst came under my observation. In these experiments 
I used the crabs E. japonicus (de Haan) from Tomioka, Tokushima 
Prefecture, which were abundantly and frequently infected with the 
cysts of the lung distome. 

In the first set of experiments I frequently found numerous cysts 
among other sediment. I have occasionally found dead crabs in the 
aquarium and pieces of legs and other parts of the body were always 
present in the material on the bottom of the vessel. It is reasonable to 
think that the cysts in the crab are easily discharged from the body 
when the crab is dead or any part of body is accidentally injured. 
Hence from observation of the above facts I believe that these free 
cysts in the aquarium were unnaturally and passively discharged from 
the body by occasion of death or some injury. 

When the crabs used in the second set of experiments were dead 
the substitute crabs were usually transported from the aquarium of the 
first set. It is even possible that the one cyst which I found in the 
second was not naturally and actively discharged from body of crab 
itself, but was carried attached to some part of the body from the 
aquarium in which I had proved the presence of cysts as stated above. 
The thickly haired forceps of the crab may be a good carrier of cysts, 
adhering to it even in case the crab had been carefully washed to 
remove attached particles. 

In considering the facts observed in the above two sets of experi- 
ments one may say that if the cysts in the first aquarium had been 
naturally and actively discharged from the crabs I should have found 
more numerous cysts in the vessels of the second series than were 
found actually. But in reality, there was only one cyst in the vessels 


178 THE’ JOURNAL OF PARASITOLOGY 


of the first set during a long time. Thus I conclude from my own 
experiments that cysts in the intermediate host (FE. japonicus) are not 
naturally and actively discharged from the body, but are often expelled 
unnaturally and passively by death of the crabs or some injury. In 
nature there are many occasions favorable for cysts escaping from 
crabs unnaturally and passively, namely, death of the crabs, frequent 
injuries by the fierce quarrels of the warlike crabs, breaking legs in 
slight disturbances, and accidental injuries in the period of moulting, 
etc. 


Japanse River Crabs which serve as intermediate hosts for Paragonimus 
westermanu. A. Sesarma dehaani M. Edwards. B. Potamon dehaanii (White). 
C. Potamon obtusipes (Stimpson). D. Eriocheir japonicus (de Haan). Photo- 
graphs by Mr. Koyama. 


LONGEVITY OF CYST IN WATER 


For studying the transfer of this cyst to a final host, it is most 
important to know how many days the cyst can be kept alive naturally 
in water. I have made the following experiments to determine this 
matter: To keep the cysts in water in a state as similar to natural con- 
ditions as possible, I prepared a small glass aquarium of 30 cm. long, 
20 cm. wide and 17 cm. deep with the bottom provided with one small 
exit. Water was constantly pouring into it by the inflow pipe and flow- 
ing out through the exit on the bottom, so the water in the aquarium 
was always moving and being renewed as in a running stream. For 
convenience in examining perfectly changes in the cysts and counting 
accurately the number of cysts dead or alive, I used as a case for 
holding them a glass tube opening at both ends covered by one or two 


YOSHIDA—ENCYSTED LARVA OF LUNG DISTOME 179 


sheets of gauze and filter paper to prevent the cysts escaping from the 
tube but to permit the water'to flow in and out though not freely. 

(A) I put forty-two cysts from the gills of E. japonicus in a tube 
whose ends were closed by gauze. The tube was placed in the aquarium 
October 30 and taken out for examination November 12, having been 
in water thirteen days. 

(B) Twenty-five cysts from the gills of the same species of crab 
were put in a tube, one end of which was closed by layers of gauze and 
two layers of filter paper and the other end by two layers of gauze 
and one layer of filter paper. The tube was kept in the aquarium from 
November 12 to 27, an interval of fifteen days. 

(C) Twenty-five cysts from the gills and muscles of the same 
species of crab were put in a tube whose ends were closed by two 
layers of gauze and two layers of filter paper. The tube was left in 
the aquarium from November 12 to December 10, or twenty-eight days. 

(D) November 15 I removed the cysts with surrounding tissues 
of the host from the gills and muscles of a specimen of E. japonicus 
that had died November 12. Twenty of these cysts were put in a tubé 
whose ends were closed as in Case C. The tube was immersed in the 
aquarium during twenty-five days from November 15 to December 10. 


The results of these experiments are listed as follows: 


Case Total Living One Dead One Cyst Only | Percentage Days 
Number 

A 26 10 (in eyst) 4 (in eyst) 4 53.8 13 
4 (outside) 1 (outside) 

B 25 4 (in cyst) 5 (in eyst) 14 20.0 15 
1 (outside) 1 (outside) 

Cc 25 (All were dead and decomposed) Bee 28 

D 20 2 | 2 | 14 10.0 25 


In Case A twenty-six out of forty-two cysts were found in the tube 
and the remaining were lost. The loss may be perhaps due to having 
closed both ends of the tube with gauze only. To avoid this defect in 
Cases B, C and D, I had used both gauze and filter paper for closing 
the tube ends, the latter being placed inside of the former. 

Cysts containing living larva were not all perfect, some of them 
being slightly broken and the others so widely broken that the larva 
was creeping out. I found there were living worms also in various 
stages. Some of them were actively moving with the light red pigment 
in the body as observed in fresh larvae, others moved slowly, and some 
others appeared dead, having no apparent motion. In the last group 
the light red pigment was greatly reduced or entirely absent. Various 
gradations of morphological change and putrefaction were observed in 


180 THE JOURNAL OF PARASITOLOGY 


dead worms. In almost all the cysts, whether the worm was alive or 
dead, swarmed an immense number of flagellata of various species. 

From my experiments above it is evident that the encysted larva 
may be kept alive relatively long in water. If a larger tube be used 
instead of a small one as in my experiments and both ends of the tube 
be closed by other suitable materials which make the circulation of 
water in the tube as perfect as possible under the conditions that retain 
the cysts, putrefaction of the cysts and their surrounding host tissues 
would be delayed and consequently the cysts would remain alive for 
longer time. Therefore we may conclude that cysts in water remain 
alive at least for thirty days under natural conditions. 

From my other experiments it is known that cysts in the crab may 
be kept alive for a week in the winter season, the gills and other 
inner parts of the crab being exposed to an air by taking off the cara- 
pace. 

METHOD OF INFECTION 


There are two ways in which the human host may be infected with 
encysted larvae from the crab: (1) by taking as food an uncooked 
crab infected with living cysts; (2) by taking with food and drink 
living cysts discharged from the crabs. Which of these two ways of 
infection is common may be quite different in various districts of the 
country, varying according to the species of intermediate host and to 
the custom of people in the district. One intermediate host, FE. japoni- 
cus, is edible and used as food in all districts of Japan, but it is gen- 
erally eaten cooked—hboiled, roasted or fried—and is rarely used 
uncooked. Another crab, P. dehaanii, is also edible and eaten cooked 
or uncooked in general. In some districts it is customary to use it 
uncooked in certain season of year. People in these districts are easily 
and commonly infected by eating uncooked crabs and a large percent- 
age of those people are found to be afflicted with lung distomiasis. S. 
dehaani is not taken as food and human infection will be brought about 
by the second method in the districts where S. dehaani happens to be 
the only intermediate host present. 

For prophylaxis in the disease caused by the lung distome the fol- 
lowing are necessary conditions: Not partaking of uncooked crabs and 
other foods washed in water in an infected district. Not drinking 
unboiled water in such district. 


REFERENCE CITED 


Yoshida, S. 1916. On the Intermediate Hosts of the Lung Distome, P. 
westermantt Kerbert. Jour. Parasitol., 2: 111-118; 1 pl. 


CYLINDROTAENIA AMERICANA NOV. SPEC. FROM THE 
CRICKET FROG * 


Minna E. JEWELL 


In the fall of 1914, while looking for parasites, I found a cestode 
in the intestine of a cricket frog, Acris gryllis. Further collections 
were made and the repeated occurrence of the tapeworm showed that 
it was not merely incidental but a regular parasite in the host men- 
tioned. This discovery was particularly interesting because of the 
rarity of cestodes, either species or individuals, in amphibians. So far 
as I have been able to ascertain, only five cestodes have yet been 
described from amphibians, and of these only two, Taenia dispar 
Goeze (1782), and Taenia pulchella Leidy (1851), are from Anura. 
No cestodes have ever been reported from a member of the genus 
Acris. For these reasons it was considered worth while to make a 
morphological and systematic study of this new form, the results of 
which are presented in the following paper. 

I wish to express my thanks to Prof. Henry B. Ward for the use 
of his library and of materials from his private collections and for 
many helpful suggestions. 

Aside from some fifty specimens I obtained from cricket frogs 
collected from a drainage ditch north of Urbana, Ill., specimens were 
also examined from Rana pipiens collected by W. W. Cort at Douglas 
Lake, Mich., and by R. G. Hall from Crystal Lake, Urbana; from 
Rana virescens collected by H. W. Duncanson near Peru, Neb., and 
from Bufo lentiginosus, locality unknown. Much of this material had 
been identified as “Taenia dispar” on the basis of its general form and 
of its host, but comparison of these specimens with specimens of 
Taenia dispar sent from Neuchatel, Switzerland, by Dr. Otto 
Fuhrmann, showed conclusively that the American worms are a dis- 
tinct species. 

Taenia dispar was originally reported by Goeze from toads and 
frogs in Germany. He described it as being 6 inches long, cylindrical, 
of greatest diameter at the anterior end and diminishing gradually to 
a thread-like posterior end. The name “dispar” was suggested by this 
unusual shape. The color is white except at the posterior end, where 
it is brownish. Proglottids are distinct only near the posterior end 
in which region they are filled with numerous brown bodies. All of 
the proglottids are enclosed in a thin transparent membrane, which is 


* Contributions from the Zoological Laboratory of the University of Illinois, 
under the direction of Henry B. Ward, No. 67. 


182 THE JOURNAL OF PARASITOLOGY 


clearly visible between the proglottids at the posterior end. Observa- 
tions were made on living material placed in water and the great 
activity of the worm noted. 

O. Schmidt (1855) studied some eighty or ninety specimens 
obtained from Rana temporaria. He pictures a worm in which the 
neck is pronounced, being about one half the diameter of the scolex 
and two fifths the diameter of the body where the testes are at their 
fullest development. Failing to find the female organs he apparently 
mistook the testes for ovaries, and while he gives us a description and 
figure of what is unmistakably an oval cirrus pouch, he fails to recog- 
nize it as such, but considers it a part of the female reproductive 
system. He describes in detail the development of the embryo and 
subsequent formation of capsules, each surrounding three embryos. 
Of these capsules he found nineteen to twenty-five in each proglottid, 
first arranged in the form of a circle, but later becoming scattered 
irregularly through the proglottid. 

Fuhrmann (1895) summarizes the contributions of previous work- 
ers and adds a careful and detailed description of his own, a sum- 
mary of which follows: 

Taenia dispar is characterized by its cylindrical form and by the 
fact that its diameter is greatest at the anterior end and diminishes 
gradually toward the posterior end. The scolex is unarmed and is not 
separated from the body by a neck. The pores are lateral and the 
cirrus and vagina pass dorsal to the longitudinal excretory vessels and 
main nerve trunk (Textfig. A). The testes are dorsal, two in num- 
ber, and measure 0.108 by 0.045 mm. The cirrus sac is a strongly 
muscular organ, having a length of 0.27 mm. and a diameter of 
0.026 mm. It terminated in a retractor which extends to the muscle 
layer on the opposite side of the proglottid. 

The female genital organs occupy the ventral part of the pro- 
glottid. The ovary is spherical, 0.081 mm. in diameter, surrounded 
by a delicate membrane and filled with forty to ninety cells 0.014 mm. 
in diameter. The vitelline gland is also spherical, but its cells are 
much smaller than those of the ovary. No shell gland was observed. 
The uterus first appears as a mass of dark cells between the ovary and 
testes. At its fullest development it is a large horseshoe-shaped organ, 
the dorsal part of which crowds the remnants of the testes against 
the dorsal muscles. The uterine wall soon degenerates and the eggs 
receive their second and then their third membranes from the paren- 
chyma. The parenchyma now becomes concentrated about groups of 
three or sometimes four eggs, enclosing them in a parenchymatous 
capsule. These egg capsules, thirteen to thirty in number, become 
scattered irregularly through the proglottid. 


’ 


JEWELL—CYLINDROTAENIA AMERICANA NOV. SPEC. 183 


It is noteworthy that there are marked discrepancies between the 
figures and descriptions of Taenia dispar, contributed by Goeze and 
Fuhrmann on the one hand, and O. Schmidt on the other. While the 
circular arrangement of the eggs described by Schmidt and the 
horseshoe-shaped arrangement described by Fuhrmann might readily 
be accounted for as differences in observation, there are more important 
differences which cannot be so readily explained. Whereas Goeze 


Figure A 
Cross section of a mature proglottid of Taenia dispar. (After Fuhrmann, 
1895) ; ¢, testes; c, cirrus pouch; d, vas deferens; 0, ovary; u, uterus; v, vagina; 
y, vitellaria. 


Figure B 


Mature proglottid of Paruterina angustata, showing the arrangement of 
organs characteristic of the sub-family Paruterinae. (After Fuhrmann, 1906.) 


and Fuhrmann both picture and describe Taenia dispar as neckless, 
having its greatest diameter at the anterior end and diminishing gradu- 
ally toward the posterior end, Schmidt, as noted above, gives a picture 
of a worm with a pronounced neck and with its greatest diameter near 
the posterior end. Further, Fuhrmann describes the cirrus sac as 
being almost ten times as long as broad, while Schmidt pictures an oval 


4 


184 THE JOURNAL OF PARASITOLOGY 


cirrus sac not more than twice as long as broad. These discrepancies 
would suggest the possibility that the form worked upon by Schmidt 
was not Taenia dispar, and that the number of taenian species found 
in amphibians is greater than has heretofore been supposed. 

Weinland (1858) attempted to put this form in a new genus, Pro- 
teocephalus, a position which subsequent workers have shown to be 
untenable. Lithe (1899) proposed for Taenia dispar the generic name 
Nematotaenia, suggested by the cylindrical form and unsegmented 
appearance of the body. Ransom (1900) gives the following diagno- 
sis for the genus Nematotaenia: 

Paruterinae; scolex unarmed without rostellum. Segmentation of strobila 
distinct only at the posterior end. Strobila circular in cross section. Genital 
pores alternate; genital canals pass dorsal to the longitudinal excretory vessels 
and nerve. Uterus horseshoe shaped, disappears early. Eggs through the action 
of numerous parauterine organs become inclosed in egg capsules, three or four 
in each capsule. Adults in amphibia. Type species Taenia dispar Goeze 1782. 

Stiles and Hassall (1912) record the following as hosts of Taenia 
dispar: Bufo americanus, Menobanchus maculatus, Bufo vulgaris, Rana 
pipiens, Rana temporaria, Ascolobates mauritanicus, Bufo cinereus, 
Bufo fuscus, Bufo lentiginosus, Hyla arborea, Necturus maculatus, 
Pleobetes fuscus, Platydactylus guttatus, Rana halecina, Salamandra 
atra, Salamandra maculata. It is likely that in some of the above cases, 
the worm found was not Taenia dispar, but the species under considera- 
tion in this paper or some form much like it. 

Taenia pulchella is known only from the rather meager description 
given us by Leidy (1851), which runs as follows: 

White, without admixture of any other color, variable, usually broadest 
anteriorly. Head quadrilateral, subclavate, obtusely rounded, broader than neck. 
Acetabula circular, cup shaped, lateral and opposite, sessile protractile. Neck 
very long, cylindroid. Articuli containing several colorless globules; anteriorly 
subglobular or transversely oval; posteriorly moniliform, longitudinally oval, ~ 
or cylindroid and centrally incrassate. Entire length, 50.8 to 238.6 mm. Scolex, 
diameter, 0.34 mm. Acetabula, 0.127 to 0.153 mm. Anterior proglottids, length, 
0.34 mm.; diameter 0.254 to 0.53 mm. Ripe proglottids, length, 0.53 to 0.57 mm. ; 
diameter, 0.19 to 0.34 mm. Host, Bufo americanus. 


Closely resembles Taenia dispar Goeze, found in Bufo viridus, etc., but it 
is relatively longer and narrower and is never colored. 


Morphology of the New Species—In making and tabulating mea- 
surements, the worms from Acris gryllis were found to fluctuate about 
a different mode from those from Rana pipiens, being usually smaller ; 
however, since every gradation in size has been found and the larger 
worms from Acris gryllis are larger than the smaller ones from Rana 
pipiens, and since also it has been observed in Acris gryllis that in 
cases of heavy infection mature worms are much smaller than those 
found in lightly infected hosts, sometimes not more than one half the 
diameter, the author feels no hesitation in saying that only one species 
is concerned. 


JEWELL—CYLINDROTAENIA. AMERICANA NOV. SPEC. 185 


Adult worms bearing ripe proglottids, from Acris gryllis, vary in 
length from 25 to 40 mm. when in a state of moderate extension. 
Young worms 1.5 mm. in length were repeatedly found in the intes- 
tine. The worms from Rana pipiens were not observed alive, but 
from preserved materials their length may be estimated to have reached 
a maximum of 80 mm. 

The most characteristic feature of the worm, noted upon a super- 
ficial examination, is its cylindrical form. The color is glistening 
white throughout the entire length. The scolex is spherical, 160 to 
200u in diameter in the region of the suckers, which have a diameter 
of from two fifths to one half that of the scolex. Thus in one scolex 
having a diameter of 180, the suckers are 97p. The neck is long and 
has a diameter of 130 to 150u. Ina typical specimen from Acris gryllis 
37 to 40 mm. in length, the neck has a diameter of 134y. The first 
appearance of the reproductive system is as a dark streak down the 
center of the worm about 5 mm. behind the head. Here the diameter 
is still 134y. 

Soon the line of undifferentiated cells becomes broken into triangles, 
having their bases directed laterad and their species alternating with 
each other in the median line. Six mm. behind the head the differen- 
tiation of the testes becomes apparent. In this region the proglottids 
have a length of 94 and a diameter of 1624. Eleven mm. behind the 
head the proglottids are mature and the first eggs are passing into the 
uterus. Here the proglottids measure 20 by 157u. The greatest diame- 
ter of the worm is found where the uterus has reached its fullest 
development and the para-uterine organ is forming, about 22 mm. 
behind the head. In this region the proglottids are 40 to 45y long and 
180 to 200» in diameter. When the worm is contracted the diameter 
may be 350u. 

Soon after, about 24 mm. from the head, the proglottids begin to 
elongate rapidly and indications of external segmentation appear. They 
now have a length of 54 and a diameter of 1354. From 27 to 36 mm. 
behind the head the segmentation is very distinct. The proglottids 
measure 82 by 108 and break off easily. The last few progtottids of 
a strobila and the detached proglottids frequently have a length much 
exceeding their diameter, 146 by 72 to 178 by 74u. In specimens from 
Rana pipiens the proglottids attain a maximum diameter of 270» and 
ripe proglottids a length of 340u and diameter of 250u. Detached 
ripe proglottids have been found singly and in groups of from two to 
five in the cloaca of the host. ‘ 

In living material the parenchymatous para-uterine organs which 
contain the oncospheres appear as two transparent spherical bodies in 
the center of each proglottid. 

The cuticula is from 3 to 4u thick, and composed of three layers, 
the central one of which is thinnest. Beneath the cuticula is the usual 


186 THE JOURNAL OF PARASITOLOGY 


basement membrane and parenchyma. The subcutaneous muscles are 
weakly developed, the longitudinal muscles are pronounced, dorso- 
ventral muscles appear to be entirely wanting. 

The ventral excretory canals vary in diameter from 3.5 to 12n, 
usually from 5 to 7 in parts anterior to the appearance of external 
segmentation. The dorsal canals vary in diameter between 1 and 4.5y. 
They are but little smaller than the ventral canals in the region of 
the scolex, but are insignificant throughout the remainder of their 
length. The usual median excretory bladder is clearly visible at the 
posterior end of young specimens. 

All of the organs of the reproductive system, with the exception of 
parts of the cirrus and vagina, are confined to the medullary region 
of the proglottid. The genital pores are lateral and alternate some- 
what irregularly, though with a marked tendency toward regularity. 
Thus in one instance twenty-four pores alternate regularly, then two 
are on the right margin and the next two on the left; then four alter- 
nate regularly, two are at the right, five alternate regularly and two 
more are at the right, twelve alternate regularly and two are at the 
left, etc. More than two pores have never been observed to occur 
successively on the same side. 

The cirrus and vagina pass dorsal to the main excretory canals and 
nerve trunk. The male organs occupy the dorsal part and the female 
organs the ventral part of the proglottid (Fig. 7). The single testis 
is situated dorsally on the aporal side of the proglottid. It varies from 
26 to 34 in diameter, being usually about 29 at its greatest develop- 
ment, and is spherical except when anteroposteriorly compressed by 
the contraction of the worm. From it the vas deferens leads with but 
few undulations directly to the cirrus. This latter organ is surrounded 
by a thick club-shaped cirrus pouch 36 to 44 long and 13 to 17 in 
diameter. The cirrus pouch, vas deferens, and female organs are 
enclosed in a delicate sheath. 

The vagina opens from the genital cloaca posterior to the male 
orifice and follows the cirrus inward. Near the end of the cirrus sac 
the vagina begins to curve ventrad. It meets the duct from the vitel- 
line gland and the very short oviduct about the level of the principal 
nerve trunks. The single spherical ovary lies in the ventral half of: 
the medullary region. It has a diameter of from 24 to 34, and con- 
tains from eight to sixteen large, spherical, loosely arranged cells 9n 
in diameter, surrounded by a membranous capsule. The vitelline gland 
lies dorsilateral to the ovary and in the median line. It is spherical, 
18 in diameter, and composed of large deeply staining cells. The 
vitelline duct passes laterad, meeting the oviduct in an enlargement 
at the point of formation of the uterine duct. No special muscular 
ootype has been observed. A mass of deeply staining cells dorsal to 


JEWELL—CYLINDROTAENIA AMERICANA NOV. SPEC. 187 


the vitelline duct is the anlage of the uterus. After fertilization the 
distal end of the oviduct becomes dilated and filled with sperm and 
yolk cells through which the egg must pass before entering the uterus. 

The ova, when mature, pass in rapid succession through the odtype 
and into the uterus so that the ovary and vitelline glands soon dis- 
appear entirely. The uterus, an oval sac, lies on the porel side of the 
proglottid with its long axis directed dorsiventrally. At its fullest 
development it attains a size of 40 by 24. The eggs at the time they 
enter the uterus may be surrounded by a transparent membrane, though 
groups of ova and yolk cells around which no membrane has yet 
formed are frequently found in the uterus. The complete eggs have 
a mean diameter of 12 to 14u. 

The parenchyma on the aporal side of the uterus now becomes 
arranged as a meshwork of heavy deeply staining strands running 
parallel to the long axis of the uterus. This is the beginning of the 
parenchymous structure which, following Fuhrmann, I shall term the 
para-uterine organ (Fig. 2). 

The growth of the para-uterine organ is rapid, and it soon appears 
as two truncated cones, one dorsal and one ventral, their bases lying 
against the uterus, which has become much flattened, and their apices 
extending almost to the circular muscles on the opposite side (Fig. 3). 
The basal portion of the cones is composed of a meshwork of fine 
dorsiventrally directed fibers. The apical parts are surrounded by 
heavy deeply staining fibers, among which lie numerous dark nuclei. 

Meanwhile the eggs have initiated cleavage and have developed their 
second membrane, a thick deeply staining capsule, while the uterus, 
which was pushed close against the eggs by the growth of the para- 
uterine organ, has broken down into a number of tertiary capsules 
surrounding the individual embryos. 

With the rapid elongation of the proglottid (Figs. 4 and 5) the 
position of the cones is shifted so that their longitudinal axes corre- 
spond very nearly to the longitudinal axis of the worm. Their apices 
lie in the anterior end of the proglottid and their basal portions, in 
which are the embryos enclosed in their uterine capsules, occupy the 
posterior part of the proglottid. At the same time the apical portions 
of the cones acquire well-defined walls and become somewhat con- 
stricted from the basal portions, while the spongy fibers which have 
filled them disappear leaving them hollow. By the time the proglottids 
have become distinctly set off, the apical portions of the cones appear 
as two thick-walled hollow spheres 20u in diameter, lying one dorsal, 
the other ventral, in the anterior end of the proglottid, while the mesh- 
work of lamellated fibers has largely disappeared from the interior of 
the basal portions of the cones. At this time the embryos have a diame- 
ter of 20 and have developed the six hooks characteristic of the tape- 
worm oncosphere. 


188 THE JOURNAL OF PARASITOLOGY 


The oncospheres now begin to migrate foreward into the spherical 
capsules of the para-uterine organ, which grow rapidly to a diameter 
of 124 to 130 (Fig. 6). At the time of the separation of the proglot- 
‘tids those embryos which have not yet migrated into the para-uterine 
capsule are usually set free by the tearing open of the end of the 
proglottid, so that a detached proglottid, when found in the cloaca of 
the host, frequently contains not more than five to seven oncospheres 
(Fig. 9). 

The development of the para-uterine organ just described bears 
many resemblances to that described for Metroliasthes lucida by Ran- 
som (1900). The chief differences are in the relative size and dura- 
tion of the uterus and the number of para-uterine organs formed. In 
the form under discussion, as noted above, the uterus is relatively 
small and breaks down into membranes surrounding the embryos long 
before the development of the oncospheres is complete or the para- 
uterine capsule is ready to receive them. In Metroliasthes lucida, quot- 
ing Ransom, “at the height of its development the uterus occupies 
almost the whole of the inner parenchyma back to the genital pore and 
bulges out the proglottid wall dorsally and ventrally,” and the uterus 
does not degenerate until the six-hooked embryos have taken up their 
final position in the para-uterine capsule. As to the number of para- 
uterine capsules formed, while in the form under discussion there are 
two, Metroliasthes lucida, although possessing a two-lobed ovary, has 
but one. 

Fuhrmann (1906) has given in less detail the development of the 
para-uterine organ in Paruterina angustata and Culcitella rapaciola, 
(1908a) of Anonchotaenia globata and (1909) of Biuterina clavulus. 
Cholodkovsky (1906) has given a brief account of the formation of 
the para-uterine organ in Rhabdometra tonuca. All of these forms 
resemble Metroliasthes lucida in that the uterus persists until the 
oncospheres have passed into the single para-uterine organ. Taenia 
dispar, on the other hand, resembles this form in that the uterus breaks 
down early, but far exceeds it in the number of para-uterine organs, of 
which there are from thirteen to thirty. 

While the form under discussion bears some likeness to Taenia 
pulchella Leidy 1851, such as its occurrence in an anuran, its long 
neck, white color and cylindrical form, this similarity is far too gen- 
eralized to establish identity. Since I have been unable to secure for 
comparison any of Leidy’s material, which is reported to be no longer 
in existence, I must leave open the question of the possible identity of 
the two forms and treat this as a new species. 

Fuhrmann (1908b) has revised the classification of the Cyclophyl- 
lides. Of his seven families it is the Dilepinidae with whose characters 
this worm agrees. The family is defined as follows: “Rostellum 


. 


JEWELL—CYLINDROTAENIA AMERICANA NOV. SPEC. tae 


usually armed, suckers unarmed, genital pores marginal, genital organs 
single or double in each proglottid.” This family contains twenty-eight 
genera, which Fuhrmann has separated into three subfamilies on the 
basis of the character of the uterus. 

The subfamily Dilepinae contains the genera in which the uterus 
is sac-shaped or has simple lobes. In most the uterus persists. The 
subfamily Dipylidiinae includes the genera in which the uterus breaks 
up into parenchymatous capsules which contain one or more onco- 
spheres. The subfamily Paruterinae includes those genera in which a 
parenchymatous para-uterine organ is formed into which the embryos 
later penetrate. The enclosure of the embryos in the para-uterine 
capsule places the worm under consideration in this paper in the sub- 
family Paruterinae. 

A comparison of the description of this form, given above, with 
the description of Taenia dispar given by Fuhrmann, reveals striking 
resemblances between the two (Fig. 7 and Textfig. A). Alike they 
are characterized by their cylindrical form and late differentiation of 
proglottids. The ovary and vitellaria are spherical and ventral, the 
vitellaria, however, being dorsal to the ovary. The testes are large, 
dorsal and of a definite and limited number, one in this form, two in 
Taenia dispar. The cirrus and vagina are dorsal to the longitudinal 
excretory canals, and the number of eggs produced in each proglottid 
is small; eight to twelve in the former, not more than ninety in the 
latter. They are further alike in that the uterus breaks down early 
before the para-uterine capsules have been formed, and in having 
more than one para-uterine organ. 

Of the other six genera of the Paruterinae, five: namely, Paruter- 
ina, Biuterina, Culcitella, Rhabdometra and Metroliasthes, are alike 
flattened dorsiventrally, the proglottids are distinct at the time of 
maturity or earlier, the female reproductive organs are anterior to 
the testes and the vitellaria posterior to the ovary. The testes are 
small, numerous (twenty to forty), and of an inconstant number, and 
occupy the posterior part of the proglottid (Textfig. B). The cirrus 
and vagina pass between the excretory canals. (In Paruterina angus- 
tata, Fuhrmann, the dorsal canal has not been observed. The genital 
ducts, however, pass dorsal to the ventral canal.) That the eggs are 
very numerous is suggested by the pictures, though no one has ever 
counted them. The uterus is relatively large and persists until the 
embryos pass into the single para-uterine organ. 

Thus it is seen that the genera of Fuhrmann’s subfamily Paruterinae 
fall into two distinct groups in one of which is Nematotaenia; in the 
other the five genera named above. The genus Anonchotaenia differs 
somewhat from either group. While the testes are small and numer- 
ous, they are dorsally situated, and the ovary, vitellaria and uterus 
are arranged laterally from the genital pore in the order named. How- 


190 THE JOURNAL OF PARASITOLOGY 


ever, this difference in the position of organs seems to be brought 
about by the shortness of the proglottid which would not admit the 
anteroposterior arrangement common in the other forms. Since, there- 
fore, Anonchotaenia is similar in general characters and in the aspect 
of the mature proglottid, and since the development of the para- 
uterine organ and of the uterus, which persists until the embryos have 
passed into the para-uterine organ, resembles that of Paruterina and 
Biuterina much more closely than that of any other form, I consider 
that Anonchotaenia is rightly placed in the subfamily with Paruterina, 
Biuterina, Culcitella, Rhabdometra and Metroliasthes. 

For Nematotaenia and the species under consideration in this paper, 
because of the pronounced differences in the aspects of the mature 
proglottids, the early degeneration of the uteri and the late formation 
of the para-uterine capsules, of which there are more than one in 
each proglottid, it seems necessary. to establish a new subfamily, 
Cylindrotaenianae: Cylindrical Dilepinidae having one or two dorsally 
placed testes, ovary and vitellaria ventral, vitellaria dorsal to ovary. 
Proglottids distinct at the posterior end only. The uterus breaks down 
early and the embryos are later enclosed in para-uterine capsules. 
Taenia pulchella Leidy would probably also belong to this subfamily. 

On the other hand, notwithstanding their marked similarity in the 
respects noted above, Taenia dispar and this new worm show certain 
important differences. As to external characters it may be mentioned 
that whereas the former has its greatest diameter at the anterior end 
and diminishes gradually to the posterior end, the latter has its greatest 
diameter about midway of the strobila and narrows toward both ends. 

Of greater importance is the difference in the male reproductive 
system. Whereas Taenia dispar has two symmetrically placed oval 
testes and the vas deferens forms a loop which passes ventrad as far 
as the excretory canals, this new worm has a single spherical testis 
situated lateral to the median line of the proglottid and a simple 
straight vas deferens. And whereas the cirrus sac of Taenia dispar 
is almost ten times as long as wide, that of the worm herein discussed 
is only two and one-half times as long as wide. 

Nowhere are the differences more striking than in the development 
of the para-uterine organs (Figs. 5 and 12) and the aspect of the ripe 
proglottids (Figs. 9 and 11) for in place of the two large elaborate 
cone-shaped structures noted above, which are probably the most 
noticeable and characteristic structures of the worm, Taenia dispar 
has a varying number of small para-uterine organs in no wise charac- 
teristic; and in place of the two large, transparent, spherical para- 
uterine capsules found in the ripe proglottids of this form, the ripe 
proglottids of Taenia dispar have from thirteen to thirty small dark 
capsules scattered through the parenchyma. 


JEWELL—CYLINDROTAENIA AMERICANA NOV. SPEC. 191 


From these considerations it becomes evident that this form does 
not belong to the genus Nematotaenia, which it most closely resembles 
of any of the genera yet established, and it is necessary to establish a 
new genus for its reception. 

This generic description would be as follows: 

Genus Cylindrotaenia. Scolex unarmed, without rostellum ; repro- 
ductive organs single in each proglottid; pores lateral, alternating; 
vagina and cirrus dorsal to the excretory canals and main nerve trunk; 
testis one, dorsal; ovary and vitellaria ventral. Uterus breaks up into 
capsules surrounding the embryos which ultimately pass into two para- 
uterine capsules. : 

Type species Cylindrotaenia americana. Characters given above. 
From small intestine of various Anura. Type specimens in the collec- 
tions of Henry B. Ward and M. E. Jewell. ' 


LITERATURE CITED 


Cholodkowsky, N. 1906. Cestodes nouveaux ou peu conus. Arch. Parasit., 
10: 332-345; 3 Taf. 

Fuhrmann, O. 1895. Die Tanien der Amphibiana. Zool. Jahrb., Anat., 
9: 207-216; Taf. 16. 1906. Die Tanian der Raubvogel. Centralbl. Bakt. Par., 
Orig. 41: 79-89; 212-221; 32 Fig. 1909a. Das Genus Anochotaenia und Biuterina. 
Centralbl. Bakt. Par., Orig. 46: 622-631. 1908b. Die Cestoden der Vogel. Zool. 
Jahrb., Suppl., 10: 1-232. 1909. Das Genus Anonchotaenia und Biuterina. II 
Das Genus Biuterina. Centralbl. Bakt. Par., Orig. 48: 412-428. 

Leidy, Jos. 1851. Contributions to Helminthology. Proc. Acad. Nat. Sci. 
Phila., 5: 239-244. 

Lithe, M. 1899, Zur Kenntniss einiger Distomen. Zool. Anz., 22: 524-539. 

Ransom, B. H. 1900. A New Avian Cestode, Metroliasthes lucida. Trans. 
Amer. Micros. Soc., 21: 213-226; 2 pl. 1909. The Taeniod Cestodes of North 
American Birds. Bull. U. S. Nat. Mus., No. 69. 

Schmidt, O. 1855. Uber den Bandwurm der Fréche Taenia dispar, und die 
geschlechtslose Fortpflanzung seiner Proglottiden. Ztschr. ges Naturw., 5: 1-13. 

Stiles, C. W., and Hassall, A. 1912. Index Catalog of Medical and Veteri- 
nary Zoclogy: Cestodes and Cestodaria. Bull. Hyg. Lab., 85. 


192 THE JOURNAL OF PARASITOLOGY 


EXPLANATION OF PLATE 


a Apical portion of Para-uterine organ / Para-uterine organ 
b Basal portion of Para-uterine organ yr Receptaculum seminis uterinum 


c Cirrus pouch s Septum between proglottids 
e Eggs t Testis 

ec Excretory canals u Uterus 

em Embryo v Vagina 

m Longitudinal muscles vd Vas deferens 

n Nerve vt Vitellaria 

o Ovary * 


All figures from camera lucida tracings except 7 and 8 which are recon- 
structions. 


Figures 1-10 Cylindrotaenia americana. 
Fig. 1—Scolex, 37. 


Fig. 2—Cross section of a proglottid with fully developed uterus and para- 
uterine organ forming, 165. 


Fig. 3—Cross section of a proglottid in the region of greatest diameter, < 160. 


Fig. 4.—Cross section of a proglottid at the beginning of external segmen- 
tation, « 160. 


Fig. 5—Toto mount, lateral view of a somewhat later stage, < 175. 
Fig. 6—Toto mount, lateral view of a proglottid near the end of the strobila, 


x 235. 
Fig. 7—Cross section of a mature proglottid, « 165. 


Fig. 8—Cross section of a somewhat later stage showing the formation of 
the uterus, < 165. 


Fig. 9.—Detached ripe proglottid, 235. 

Fig. 10—Frontal section of three proglottids in the region of the greatest 
development of the testes, 325. 

Figs. 11 and 12—Taenia dispar; from materials sent from Neuchatel, 

Switzerland. 

Fig. 11—Toto mount, ripe proglottid. Stage corresponding to Figure 9 in 
Cylindrotaenia americana, < 50. 

Fig. 12—Toto mount, proglottid from near the end of the strobila showing 
para-uterine organs. Stage corresponds to Figure 5, 70. 


PEATE 


C--*-—% es 
. ee 


ned oy SS 


THE *EFPEGT* OF CTI’ BITES ‘ON MAN 


D. McCAFFREY 
PRINCETON, B. C., CANADA 


The local and constitutional effects which tick bites have on human 
beings is a subject which is still in the experimental stage, and I have 
been unable to find it discussed at any length in any of the textbooks. 
Osler merely mentions the fact. The only literature available is to be 
found in papers published in the scientific magazines. 

I have had several cases which I attribute to the bites of ticks. 
These cases present two sets of symptoms, that is, local and constitu- 
tional. Where the local symptoms appear the tick has been forcibly 
removed, and the parts which it buries into its host left behind. The 
constitutional symptoms appear when the tick is allowed to remain 
on the host for some time. To illustrate the local symptoms I will 
describe two cases. 


Case 1—M. M., aged 34. One night in May, 1913, on retiring, he felt an 
irritation along the right tibia. He rubbed the leg vigorously with the hand. 
Next morning the pruritus was very marked and a red spot appeared on the 
leg. The body of the tick was found on the floor. That day the leg began 
to swell and continued to do so until it was nearly twice its normal size. An 
abscess developed at the point where the tick’s head was embedded. Hot 
poultices were applied for a week, then an incision made. The skin over the 
abscess was very tough. A creamy looking pus escaped. In about three days 
a hard dark colored mass came away. This was almost of a rubbery nature, 
and left an ulcer which extended nearly to the bone. I treated this as an 
ordinary ulcer for a week, but it showed no signs of healing. By that time 
the ulcer had hard indurated edges, giving it a “punched-out” appearance. There 
was a thin watery exudate coming from it. The only thing the patient com- 
plained about was the intense itching. In two months the ulcer was covered 
with skin. Each spring an ulcer has formed at the same place, which takes 
from two weeks to two months to heal. The leg itches almost constantly, at 
times becoming almost unbearable. 


CasE 2.—Female, age 9, June, 1915. The tick was found on the mastoid bone. 
Her father removed it by force. Next morning the child complained of itch- 
ing, and a red spot appeared. In two days the parts were much swollen and 
tender, with a raised spot where the head was imbedded. Applied hot poultices 
for four days and incised. The skin was very tough and very little pus 
escaped. The next day pus came from the ear as well as by the incision. On 
the third day after the incision was made, a dark, rubbery mass came away, 
leaving a “punched-out” ulcer. The ulcer has not healed as yet, Aug. 15, 
1915, the pruritus being so severe that the child’s hands have to be tied at night. 


To illustrate the constitutional symptoms I will describe the only 
case I have had. 
Case 1—D. W. Female, aged 11. May, 1915. Retired May 9, in ordinary 


health. When she got up the next morning her legs gave way and she fell. 
She walked about that day with no other symptom than falling, if she turned 


194 THE, JOURNAL -OF PARASTTOLOGY 


or moved quickly. She could execute any movement if done slowly. The next 
day she could not walk and her arms were involved. Paralysis gradually 
extended until all the muscles were involved, leaving the child helpless. The 
pupils gradually dilated and lost their power of reacting to light. The tendons 
were at first exaggerated but later became lost. For the first four days the 
patient was very excitable. The muscles twitched so as to give her choreic 
movements; afterward she became somewhat duller but still retained most 
of her faculties. The involuntary muscles were the next to become affected, 
so that there was incontinence of the urine and feces. The breathing which 
was at first rapid became “choky,” there being a peculiar rattling sound at 
each effort to breathe. She complained of a lump in her throat. If given 
liquids they returned by the nose. Her speech was affected so that she could 
hardly articulate. The tongue became swollen. The heart became very rapid, 
being above 120 per minute. The temperature at first rose 1 degree then 
dropped to 3 degrees below normal. Urine analysis negative. Sensory nerves 
normal. On the seventh day after symptoms appeared I removed a tick from 
near the crown of the head. Recovery was very rapid, so that on the third 
day she was able to walk up the street. In this case I had practically given up 
all hope of recovery. When the tick was removed I stopped all medicine and 
treatment. The child made a complete recovery. 


I was fortunate enough to have the tick in this case identified as 
Dermacentor venustus. Whether it is Dermacentor venustus which 
causes the local effects or Dermacentor albipictus, which is also quite 
plentiful on horses in this district, | am unable to state. Princeton has 
an altitude of 2,000 feet and ticks are most abundant and active dur- 
ing the spring months and early summer; possibly they may be found 
higher up in the mountains at a later date. The case of “tick paralysis” 
I have just described is the first one of its kind that I have seen in the 
Princeton district. A number of cases have been reported from other 
parts of British Columbia, the nearest being in the Similkameen Valley 
some miles distant. 

Dr. S. Hadwen examined the tick which was removed from the 
case of “tick paralysis,” and determined it as Dermacentor venustus, 
a half-gorged female. He tells me that he has never seen any harmful 
effects from the bites of D. albipictus in animals, but that the local 
after-effects from the bites of D. venustus are often severe. Accord- 
ing to Hadwen, the constitutional effects in animals following the pro- 
longed attachment of D. venustus are identical to those I have just 
described. 


SOCIETY PROCEEDINGS 


THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON 


The twenty-eighth regular meeting of the society was held at the residence 
of Mr. Crawley Dec. 10, 1915, Mr. Crawley acting as host and Mr. Chambers 
as chairman. Dr. Cobb presented the following: 


Notes on New Generaand Species of Nematodes——Note 1—Antarctic Nematodes 


The free-living marine nematodes of the Mawson Antarctic Expedition rep- 
resent twelve species, nine of them new (one new genus), and three species 
previously described in my report on the free-living nematodes of the Shackleton 
Expedition. This raises the number of known marine species of Antarctic 
free-living nematodes to thirty-four, representing eighteen genera, only three of 
which are new. Considering the small number and the meagerness of the 
Antarctic collections, these results indicate that Antarctic species of marine free- 
living nematodes are very numerous and belong to very widely different genera, 
and for the most part to genera found in warmer seas. 


Note 2.—Renette of Cephalobus 


I find that in some species of Cephalobus, and probably in the majority, the 
excretory or renette duct is bifurcated and passes along the lateral fields to 
near the posterior end of the body. This structure thus parallels that found 
in many species of Rhabditis, and as a considerable number of parasitic forms 
have either rhabditiform larvae, or rhabditiform free-living generations, the 
possibility is suggested that Cephalobus, or rather some species of it, may be 
free-living forms connected with parasites. Examination of a large number of 
marine free-living nemas (a contracted term proposed here for the word nema- 
todes) has strongly impressed me with the possibility that some of these species 
will in the end prove to be free-living forms of parasites of fishes, marine birds, 
cetaceans, etc. 


Note 3—A New Form of Nematode Hermaphroditism 


I have a new nematode species that is extremely interesting in the form of 
its hermaphroditism. The individuals have the form of females, with double 
sex organs, one of normal size and functioning as an ovary, the other exceed- 
ingly small, and appearing to function as a testis. 


Note 4.—Subdivisions of Mononchus 


I find the free-living nematode genus Mononchus Bastian, 1866, to be divis- 
ible into five very natural divisions, of which the first three form a group 
considerably differentiated from the two others which may later be raised 
to the rank of genera. 


1. Mononchus typical subg—Pharynx twice to thrice as long as wide; onchus 
massive, midway or farther forward, unopposed by denticules ; pharyngeal walls 
smooth or transversely striated; males of six species known; ovaries, two, 
reflexed. Type species M. truncatus Bast. Consisting of such species as M. 
M. brachyuris Biitsch.; M. parvus de Man; M. rex Cobb; M. fovearum Dyj.; 
papillatus Bast.; M. intermedius Cobb; M. major Cobb; M. gerlachei de Man; 


196 THE JOURNAL OF PARASITOLOGY 


M. macrostoma Bast.; M. longicaudatus Cobb; M. tunbridgensis Bast.; M. 
dadayi Micol. 

2. Prionchulus subg. nov.—Pharynx about twice as long as wide; onchus 
massive, midway or farther forward, opposed by numerous denticules arranged 
along a longitudinal pharyngeal rib; males of one species known; ovaries, two, 
reflexed. Type species Pr. muscorum (Duj.). Consisting of such species as 
Pr. muscorum (Duj.) and Pr. spectabilis (Ditlevsen). 

3. Mylonchulus subg. nov.— Pharynx goblet-shaped; onchus more or Iess 
arcuate, massive, midway or farther forward, opposed by numerous denticules 
arranged in transverse rows on two rasp-like areas; males unknown; ovaries, 
two, reflexed. Type species My. minor Cobb. Consisting of such species as 
My. minor Cobb and My. obtusicaudatus (Daday). 

4. Iotonchus subg. nov. (gen. noy.?).—Dorsal onchus and all others usually 
basal, relatively small; large species with large, elongated pharynx, having 
three longitudinal ribs; tail rather long and slender; males of two species 
known; ovaries, one or two, reflexed. Type species I. gymnolaimus Cobb. 
Consisting of such species as J. digiturus Cobb; J. bathybius (Micol.); I. 
studeri (Steiner) ; I. tridentatus (de Man). 

5. Anatonchus, subg. nov—Onchi retrorse, midway in pharynx or sub-basal; 
large species with roomy elongated pharynx; tail long and usually becoming 
cylindroid; female organs double; males of most of the species known. Type 
species A. tridentatus (de Man). Includes A. dolichurus (Ditlevsen). 

I have manuscript descriptions of several new mononchs from various parts 
of the world, all readily referable to one or another of these divisions. 


Note 5.—Finder Slides 


In an article in the Transactions of the American Microscopical Society 
(34:1-89) I have suggested the advisability of using co-ordinate numbers, 
preferably probably minus co-ordinates, dating from the upper right corner of 
the slide as the origin, or zero point. The slide I am exhibiting is of this kind, 
and presents the peculiarity that it does not have to be constantly removed and 
replaced when in use, thus effecting a material saving in time and energy. It 
consists of a series of coordinates arranged in a small holder adapted to receive 
and clamp the microscope slide upon and in register with the finder. Light 
from the microscope mirror passes through the finder and the microscope slide. 

In other words, the finder slide is ruled into millimeter squares, each square 
containing two numbers indicating the actual distance of the square from the 
right-hand edge of the slide and from the top of the slide, respectively. Under 
the microscope the normal inversion makes these numbers appear to read from 
the left-hand side and from the bottom of the slide. The slide which is being 
studied fits over the finder and is held by two small fixed clamps. By focusing 
down at any point the two indicative numbers for the corresponding square 
may be found and noted. The slide is made by photographing a ruled and 
numbered sheet with such a reduction as will make the photographic squares 


one millimeter square. 


Dr. Stiles presented a note in regard to the sanitary index of three Southern 
communities, A, B and C. In two of these communities, A and C, the authori- 
ties in charge had preached what they regarded as feasible, but comparatively 
low, standards of sanitation, including the advocacy of the unsheltered or 
so-called “umbrella privy.” In the third community, B, the authorities had taken 
the position that it was not advisable to advocate something that would have 
to be combated subsequently, and in consequence high, even if temporarily 
unattainable, standards had been advocated. After the lapse of a year, the 
sanitary index of the three communities was again taken and compared with 
the index for the period of the sanitary campaign of a year before. It was 


. 


HELMINTHOLOGICAL SOCIETY OF WASHINGTON 197 


found that the sanitary index for the two communities A and C had fallen in 
a year from 28.1 to 24.2 for A, and from 34.6 to 29.4 for C, while the sani- 
tary index had risen for community B from 31.5 to 45. The sanitary campaign 
in communities A and C was of the revival type with much attendant publicity; 
that of community C was of a quiet, personal nature without so much attendant 
publicity. It was found that the umbrella privies built in communities A and 
C had gone to pieces in a year. 


Dr. Stiles also presented a note on memory span studies in children. Of 
children from homes with privy and those from homes with sewer, it was 
found that the memory span of the last group compared with that of the 
first group as 14 to 10. For thirty-six boys and sixteen girls with light infesta- 
tions with hookworm, the total memory span should have tested 343.24, and 
did in fact test 339, showing only a very slight variation below normal. For 
thirty-eight children infested with Ascaris, the total memory span should have 
been 245.23, and was in fact 250, a slight variation above the normal. For 
sixty-seven children infested with Giardia (Lamblia), the total memory span 
should have been 441.6, and was in fact 444, a slight variation above the 
normal. For fifty-five children infested with Entameba coli, the total memory 
span should have been 367.29, and was in fact 376. It therefore appears that 
while children from sanitary homes show a superiority over those from insani- 
tary homes, so far as the memory span is concerned, of 14 to 10, the presence 
of slight infestations with hookworm, ascarids, Giardia or Entameba coli appear 
to bear no appreciable relation to the memory span. 


Maurice C. Hatt, Secretary. 


The twenty-ninth regular meeting of the society was held at the residence 
of Dr. Stiles Jan. 28, 1916, Dr. Stiles acting as host and Dr. Pfender as chairman. 

Dr. Stiles presented a note in regard to cases of spurious parasitism. A 
slug, said to have been passed by a patient in Baltimore, and identified by 
Dr. Paul Bartsch as Limax flava, was shown to the society. In a second case, 
a physician had for years been regarded as presenting a case of multiple infesta- 
tion with Cysticercus cellulosae, this being the diagnosis of the patient and of 
several other physicians. A physician who had examined the patient called in 
Dr. Stiles, and their examination disclosed the fact that the patient was addicted 
to the use of drugs administered by the usual hypodermic method. The patient’s 
failure to us a properly sterilized needle had led to the formation of the small 
swellings which were present over the arms, legs and the portions of the 
body accessible to the needle, but significantly absent over the back. These 
swellings constituted the supposed cysticerci. One of these swellings when 
excised and sectioned showed connective tissue and pus. Dr. Stiles also noted 
the fact that the pulp vesicles of an orange had been sent to him with a diag- 
nosis of Dicrocoelinm lanceatum, and predicted that next spring and summer 
there would be the usual amount of hairs from the strawberry sent in as 
supposed specimens of pinworms and hookworms. He also recalled the send- 
ing in of a specimen, said to have been vomited by a boy and supposed to 
be parasitic, which proved to be a placental structure, apparently from a cat, 
and called attention in this connection to the historical Spiroptera hominis, 
which had proved to be the entrails, eggs and encapsulated nematode parasites 
of fish, which had evidently been introduced into the vagina by a hysterical 
woman patient. - 

Dr. Ransom presented the following notes on spurious parasitism: There 
was at one time in Washington a man who was accustomed to come to the 
Bureau of Animal Industry with an account of a peculiar affliction consisting 
in his being parasitized by insects which would suddenly appear in the skin, 


198 THE JOURNAL OF PARASITOLOGY 


quickly emerge and fly away. The man appeared sane on other topics, Dr 
Ransom also noted a case in which supposed flukes were sent in as having been 
vomited by a boy. Examination showed them to be earthworms. In another 
case of a similar nature the supposed parasites proved to be two earthworms 
and a slug. 

Mr. Crawley noted a case in which blood smears were sent in with the report 
that they showed blood parasites. These objects proved to be a common 
fungous structure which occurs in feces of all sorts almost anywhere. 

Dr. N. A. Cobb gave a stereopticon demonstration, discussing about thirty 
species of nematodes found in the sand of slow filter beds from the filtration 
plants of various cities, and presenting three notes thereon: 


Notes on Filter-Bed Nematodes—Note 1.—Predaceous Nematodes 


The discovery of nematodes in tap water led me to an investigation of 
conditions at filtration plants. Nematodes were found on the walls wet with 
spray at the flumes where the filtered water enters the city’s supply. At the 
end of the period of use, usually a few weeks, the sand in the beds was found 
to contain hundreds of millions of nemas per acre in the top 3 inches. In 
one case, where the tale reached about one thousand million nemas per acre, 
nine tenths of the specimens were of one species, the predaceous Mononchus 
longicaudatus Cobb, which feeds on other nematodes, protozoa, etc., and hitherto 
known only from soil. This species is cosmopolitan. Another mononch, the 
Mononchus papillatus Bastian, I have shown, feeds on the citrus-root nema, 
an injurious parasite of various citrus trees, and there is a possibility that the 
filter-bed form may be economically serviceable in destroying injurious nemas. 
The filter-bed form is interesting from the fact that good preparations show 
that the esophagus is supplied with glandular structures opening into the lumen. 


Two vegetarian species of Monhystera were found in the filter beds feed- 
ing on microbes and other organisms, and a species belonging to a new genus 
has the same food habits. . Jronus ignavus Bastian and Jronus longicaudatus de 
Man, also found in large numbers in the filter beds, show in the cells of the 
intestinal walls doubly refractive granules which have also been found in the 
lumen of the intestine, indicative of a cannibalistic food habit. J. ignavus has 
an interesting egg, with peculiar chromatic elements scattered through its cyto- 
plasm. In both forms the renette, hitherto undiscovered, is well developed and 
empties near the lips. Both have esophageal (salivary?) glands emptying into 
the pharynx. 

Tripyla monhystera de Man is a very active, rapacious, carnivorous nema 
feeding on other nemas and on rotifers and protozoa, and is very common in 
filter beds. It suffers from what appears to be a protozoan disease, the 
protozoan usually invading it in the region of the tail, the invasion progressing 
most rapidly along the lateral fields. The affected nemas lose their normal 
activity and show signs of disease. The infection terminates, at least at times, 
in the death of the host. 


Note 2.—Syngonism and Parthenogenesis; Cryptogenesis 


Among these filter-bed nemas I have quite a complete series from bisexual 
species, through those showing obvious syngonism with prominent development 
of sperm in the gone followed by egg development, to those syngones in which 
the sperm development is rapidly accomplished and results in relatively incon- 
spicuous though functional sperms. So complete is this series, ending in 
sperm discoverable with the utmost difficulty on account of minuteness, that 


HELMINTHOLOGICAL SOCIETY OF WASHINGTON 199 


the fact that in any particular case the presence of sperm was not demonstrable, 
as, for instance, was the case dm a species of Ironus, could not be regarded as 
proving its absence. Since in syngonism there is a single primordial gonic 
cell which by division gives rise to sperm and then to eggs in the same gone 
within a very short time, the idea is suggested that instead of this cell division 
producing these various elements and then a little later uniting them in the 
process of fertilization, the essential processes might occur in the earlier uni- 
cellular stage and the whole affair be consummated as a more nearly simul- 
taneous instead of a consecutive process. This theory I suggest for considera- 
tion in connection with parthenogenesis. Such a method of reproduction, if 
it exists, I would denominate cryptogenesis. 


Note 3.—Revtsion of the Genus Cylindrolaimus 


Careful examination of a new species of Cylindrolaimus from the Wash- 
ington filter beds has led to a more complete characterization of Cylindrolaimus, 
and a revision of the genus, as follows: 

Cylindrolaimus de Man, 1884——Small squatic or meadow-land species, with 
naked, striated cuticle; cephalic setae, four, spreading, submedian; pharynx long, 
narrow, cylindrical, unarmed; lips rudimentary or none; labial papillae exceed- 
ingly minute; amphids circular, depressed; esophagus cylindroid, valveless, with 
well-developed cylindrical cardia; intestine thick walled, granular, not tessellated; 
tail moderately long, usually blunt, containing three caudal glands emptying 
through a plain, rounded, unarmed spinneret. Ovary single, outstretched; with 
a small branch on the other side of the vulva. Males rare or none, and, so far 
as known, having two equal, arcuate spicula, with very rudimentary accessory 
piece; male supplementary organ one, simple, slightly elevated, opposite the 
spicula; C. communis de Man denominated type species by de Man. 


Key to Species Thus Far Referred to Cylindrolaimus 


The last species (5, 6, 7 and 9) are not cylindrolaimi; the genus to which 
each may belong is suggested in parenthesis: 
Bulb about pharynx, none; ovary one (except 

in No. 6); tail simply conoid; head rounded; 

ampiidseas wide as Pharyix. =o. 00+ secs f-communis de Man 1 


Amphids half as wide as pharynx; ceph. setae 
half long as head is wide; oes. 20%; spin. 


SMITE ACME 6 0 po ope wafers 1p we erase daria -f obtusus n. sp. 2 
Ceph. setae papilloid; oes, 14%; spinneret asym- 
AGEETED ren vee. cibie ns co aloohe Shae so mis ite eGo ote ? melancholicus de Man 3 


Tail conoid, then cylindroid; head more or less 
truncate; pharynx twice long as head is wide; 
ceph. setae four or none; uterus and ovary 
simple; amphids minute or none; ovary re- 
flexed; amph. small entering obliquely 


Ses Siscis-s Sbtk (Cylindrolaimus ?) ‘f tristis Ditlevsen 4 
Ovary outstretched; no amphids, setae or spin- 
eS a ne ci An eS (Gen. nov.?) -f macrurus Daday 5 
Uteri 2; ovaries reflexed; amphid a spiral 
as aoe Seeecerrr ee oe i. eo (CP lectus?) ‘f’ aberrans Micoletzky 6 
Pharynx as long as head is wide; cephalic 
Setae "Gh a. eee a. (Prismatolaimus ?) -f politus Daday 7 
EES TE RRS SS aay An 8 i) SS OS -f brachystoma Hofmanner 8 


Bulb of pharynx distinct, ovaries 2; setae none 
SCI ete Pape ee ees ee (Ethmolaimus ?) ? lacustris Hofmanner 9 


200 THE JOURNAL OF PARASITOLOGY 


a ee ee F 

C. obtusus n. sp. *~ 20 26° ~ 33 “35 ~~23’ 6mm. Resembles C. communis, 
from which it differs in the form of the female sexual organs, the cephalic 
setae, and form and size of the amphids. Ventral excretory pore opposite the 
middle of the pharynx. Appears to be digonic, since the small outstretched pos- 
terior branch of the sexual organ appears to function as a testis. Habitat: 


sand-filter beds, Washington, D. C. 


melancholicus — tristis communis —macrurus 


ae 


ANSON NIT 


Fig. 1—Heads and tails of species of Cylindrolaimus referred to in the key, 
reproduced from illustrations in the published descriptions of the species. 


ye “ ; AE eis ea | Pte we 
melancholicus (-2- \j-----“'-~*33- 2° 3 mInchlicus (= \j-—-r ~~ 3-38 28° “™ 


f= R I 9. 562! 88.5 - . eds ? 265 nd Coe 
commums isis Se em gsiis =n tt oo 
ee ee ee) 
macrurus 3} 82 sae aborrans Ete 9y 2 ptm 
. 7eaeeG{. <216;  <60:. 1 56F ey one | ae Mel 2 ee eS 
politus (as ee Jocustns (= 2 


Fig. 2—The formulae of the species referred to in the key. 


Dr. Pfender presented a note in regard to a patient who thought that he 
had a tapeworm. Radiographs presented by Dr. Pfender showed that the 
symptoms which the patient referred to were due to nephrolithiasis. A nephrec- 
tomy was performed and the stones, one large one and numerous smaller ones, 
which had been found in the kidneys, were exhibited. 


Maurice C. Hatt, Secretary. 


REVIEWS AND NOTES 


DIE TIERISCHEN PARASITEN DES MENSCHEN 


1. Tem: NATURGESCHICHTE DER TIERISCHEN PARASITEN DES MENSCHEN VON 
Dr. Max Braun. Fiinfte, vermehrte und verbesserte Auflage. 560 pp. 407 
text figures. Curt Kabitzsch Verlag in Wiirzburg. 1915. 13 mk.; geb. 15.50 mk. 


The appearance of a new edition of the well-known and highly prized text 
by Braun is deserving of more than ordinary notice here. It is seven years 
since the fourth edition was published and the literature on parasitology which, 
especially among Protozoa, has modified and extended the world’s knowledge of 
this important field, may fairly be said to have doubled in that short interval 
of time. The last (fourth) edition of Braun’s work saw the addition of a 
clinical-therapeutic section which in this edition has been expanded to a second 
part, the separate appearance of which is promised at an early date. Unques- 
tionably it was the rapid growth in materials demanding consideration which 
has led to the separation of the newly introduced section as an independent item; 
for the first part, which covers only the structure, life history, distribution and 
classification of the species parasitic in man, now utilizes 560 pages—or more 
than twice the compass of the entire work in the third edition. 


The increase in size is also accompanied by marked changes in form such 
as to allow of more extended discussion in the same space. The type page is 
both larger and wider. More of the present volume has been thrown into fine 
type and other means of condensation have been employed freely in the effort 
to bring present knowledge into a reasonable compass. Unfortunately in this 
process the author was compelled to reduce the introduction considerably in 
extent, a change which every one must view with real regret for Professor 
Braun is an artist in presenting concisely and clearly any discussion of general 
principles and many generations of students have read with profit and delight 
his opening chapter on Parasitism in General. The general discussions with 
which the account of each group was introduced have also suffered somewhat 
in the process of condensation. 

Even with all this trimming the text has grown fully twenty per cent. in 
volume. New material is in evidence everywhere. The new edition is in fact 
a real revision and not a mere reprinting with minor textual modifications. The 
author has added a very considerable number of new species which have been 
discovered since the appearance of the last edition or which since then have 
been found to be of significance to man. This increase runs from ten to thirty 
per cent. in different groups. 

The plan adopted in the previous edition of grouping the important references 
to the literature in a section at the close of the text proper has been followed 
here more consistently. This list, though sharply scanned, has increased greatly 
in extent and now covers 110 text pages. It is notably fairer than most foreign 
lists in its treatment of American work and is thoroughly up to date. A few 
typographical errors were noted and some curious abbreviations in titles of 
English articles. Unfortunately the references on topics in the last 25 pages 
are printed in the text after the manner of earlier editions and not brought 
together in the bibliographic list. This detracts somewhat from the character 
of the work and the record is also not so good. Yet, one may say confidently 
that it is the best reference list available in this field. 


The illustrations are frequent, good, and about one fourth of them new. 
Some ancient favorites that are not very accurate still occupy their historic 


202 THE JOURNAL OF PARASITOLOGY 


places. Thus the figures of Demodex and of the female /xodes ricinus are 
little worthy of a place in such a work. But on the whole the work is better 
and more profusely illustrated than our own texts in biological science. 

The section on Protozoa has perhaps been modified most of all. The system 
is greatly expanded and one notes that the Cnidosporidia have been exalted to 
the rank of a class, a conspicuous departure from the time honored division of 
this phylum into four classes. By the removal of the discussions concerning 
insect vectors (mosquitoes, biting flies, etc.) to the chapter on insects the 
apparent increase in size is not marked superficially, but the space gained in 
this way is more than filled by data on the group proper. New species, new 
data on morphology, life history, and biology, as well as recent experimental 
work, and new figures are prominent in this section. 

In the chapter on Trematodes the author has introduced a systematic outline 
prepared by Odhner and embodying the recent important researches of that 
distinguished investigator on the relationships of the various groups of flukes. 
This system marks a distinct advance in the direction of a natural classification 
based on comparative anatomy and follows the line of attack formulated in 
Looss’ epochal studies on the natural classification of the Trematodes. It is 
interesting to note that even in this long known and much studied group, the 
text lists seven species out of twenty-one that were not mentioned in the 
previous edition, and that the accounts of species formerly listed almost all have 
been radically revised in correspondence with recent discoveries concerning them. 
The author displays commendable conservatism in refusing to follow extreme 
modifications in nomenclature and yet he has not hesitated here or elsewhere in 
the volume to use new names when their establishment rests upon adequate study 
and morphological demonstration. 

Among the Cestodes which furnish the fewest species to the list of human 
entozoa there are less changes to record. The older species have undergone 
little alteration though one name, Hymenolepis lanceolata, has been eliminated 
on the basis of error as indicated by Fuhrmann in 1908 in a note generally over- 
looked. But it will surprise even those somewhat familiar with the literature 
to find that among the twenty-seven species of tapeworms listed in this work 
seven are new within the last seven years. 

No group of helminthes has undergone greater changes in recent years and 
is still in greater need of revision on the basis of such studies as Looss, Lihe, 
and Odhner have made among Trematodes, than the Nematodes. Thanks 
especially to Goldschmidt, our knowledge of nematode structure has been greatly 
advanced and full use is made of this advance in the work under review. The 
system is still a disconnected series of “families,” based on factors very dis- 
similar in character and value. representing thus sometimes a small group of 
closely related species and in other cases a large mass of anatomically variant 
forms drawn together by artificial definitions. Some slight progress has been 
made in the solution of the difficulties by the pioneer work of Railliet and Henry 
which has been used by Braun. Nevertheless the “system” remains little more 
than a list. Under individual species Braun has added much new and important 
material, such as the work of Fulleborn on filarial life history, of Looss on the 
hookworms and other species, and of many others. A good many new species 
and also some new names greet the reader in this section. Fortunately figures 
and descriptions are now adequate in the main for an understanding of the 
species and for their differentiation; this which has not been true in earlier 
works, will do much to clear up the confusion which exists in this group. 

Among the Arthropoda (mostly ectoparasitic mites and insects) one notes 
a fuller treatment which conforms to the now fully demonstrated role of these 
forms in the transmission of protozoal diseases. The increase is due partly, as 
already noted, to the transfer of such materials as in earlier editions were found 


REVIEWS AND NOTES 203 


under other headings. This change has made distinctly for the unity and clarity 
of the work and was-necessitated further by the demonstrated agency of the 
single form in the transmission of various parasitic organisms, e. g., the mos- 
quito as the inoculator of several protozoa and filariae. But beyond this the 
arthropod section contains much new material. Especial mention should be 
made of the fine new figures and the carefully collated data for differential diag- 
nosis of important species. Of course the accumulated knowledge in this part of 
the field is great, as exemplified by recent works devoted exclusively to it, and 
Braun has not attempted to include all. But as a summary this part must be 
recognized as a real success and a great advance over the distinctly inadequate 
treatment accorded this phase of the subject in earlier editions. 

All in all the new edition represents a most valuable contribution to helmin- 
thological literature. It is a worthy production of the famous head of the 
K6nigsberg school of parasitologists and justly entitles him again to the con- 
gratulations and thanks of other workers in this field. 

Doctor Jesus Rafael Risquez of Caracas has published an interesting study 
of nineteen cases of the blood fluke (Schistosoma mansoni) observed in eighty- 
six autopsies in Venezuela, fourteen of which came from the white race, none 
from the indian or negro, and five from half breeds; in large part the patients 
were born in Caracas. 

The Report of the United Fruit Company’s Medical Department for 1914 
was reviewed in this Journal-last December. The Report for 1915 confirms in 
essential details the conditions regarding the occurrence of human parasites on 
the shores of the Caribbean which were taken from the previous report and 
embodied in the tables of the review cited. . 


HIBERNATION OF MUSCA DOMESTICA 


In 1913 Dr. Henry Skinner challenged the commonly accepted belief that 
adult house flies remained dormant throughout the Winter months. He even 
went so far as to say tentatively that house flies passed the Winter in the pupal 
stage and in no other way. Dr. Johannsen’s observations at Ithaca tended to 
confirm Dr. Skinner’s conclusion insofar as it applied to conditions in the 
latitude of New York State. 

In January of this year an instructor in the Department, Mr. W. L. Chandler, 
observed several adult specimens of Musca domestica in the sub-basement of 
Roberts Hall, one of our University buildings. I have observed others in the 
sub-basement and around in the buildings even at this late date (April 7). 
These were remote from breeding places and there seems no possibility that 
they hibernated in the pupal stage. Witiiam A. RILEY 


In a recent important paper Crawley has shown that when mice are fed 
material containing the so-called spores of Sarcocystis muris invasion of intes- 
tinal epithelial cells by the parasites takes place withintwo hours. This phenom- 
enon is most favorably studied in the last inch or two of the small intestine. 
Within the cells, the parasites rapidly separate into two categories, the latter 
history of which shows them to be males and females. 

In the male, development takes the form of a notable increase in the size of 
the nucleus, correlated with a loss of most if not all of the cytoplasm. Various 
internal changes take place within this enlarged nucleus, and eventually the 
chromatin becomes divided into clusters of minute granules, grouped around 
the periphery. These granular clusters solidify into compact balls, which 
elongate and produce the microgametes. 


204 THE JOURNAL OF PARASITOLOGY 


In the females, the changes are not so conspicuous. The cell becomes 
shorter and broader than the original spore, but there is no loss of cytoplasm 
nor any conspicuous enlargement of the nucleus. The nuclear chromatin remains 
concentrated in a large karyosome. 

This sexual evolution is completed in from 9 to 18 hours, after which 
fertilization takes place. The further history of the zygote has not been followed. 


205 


INDEX TO VOLUME II 


PAGE 
Acanthocephala from Fresh-Water Hosts, Seasonal Distribution of Some.. 106 
Arachnoid, Pneumonyssus foxt nov. sp., Parasitic in the Lung of a Monkey 


CRERCOCUS SHCGUS) cite tarde ace eee ae ata che al sia'a'a"s'e'e'e'e'e'n's’s ou na d's 
PGenives) lee rarasnoleeie (Hele) ue, we eee eos. clo. e lees ed cwand 148 
Agchivos Brasileiros dé Medeeinia (review ) ce. 202 ccs ceidc nese cc cen covecek 148 
Are Sarcosporidia Aberrant Forms of Cnidosporidia of Invertebrates?.... 126 
Arhythmorhynchus, Revision of the Genus, with Descriptions of Two New 

Species Trans INOriE siiterieanl ite ns ees SOs shail vis .ccesecade 167 
Arthropoda, Some New Gregarine Parasites from...................2008. 27 


Barrett, M. T., see Smith, Allen J., and M. T. Barrett. 

Bartonella, Note on the Etiology of Verruga as Deduced from a Study of 
ENEMAS RUALMNS EASES HOt le igen ae en eI eel Nn clas ait stoi seers cibdid © ties 6 143 

Book Reviews, see Reviews. 

Braun, Max: Naturgeschichte der tierischen Parasiten des Menschen, 1. 
ERS G eure. rie Na I BA SO eS ee rn 201 

Brazile. Medical Zoology, ail (DOO LEVie ws) ir cia as tosis cisig cle iwic-c)oistec cio sees 148 

Catostomus commersoniu, On the Occurrence of a Trypanoplasm, Probably 
Trypanoplasma borreli Laveran et Mesnil, in the Blood of the Com- 


AIG SEE el weytelsleiayal ee sties rose satetoler sears e wielecte otstle cioxe op erese/ain ie. elle ast vce sve 1 
Cattle Tick, Margaropus annulatus, Note on the Stage of Piroplasma 
RCCIRN ee GION OCEMNS M6 EMEA dine aa Satin ag has chan veee as secs ects 87 
aes Ses Teele WISint at oe an wad Gree aca een Fito lstid De yn ccceccece 46 
Cestodes, Polyradiate, Two New Cases of, with a Summary of the Cases 
PAA PICO IRG laces ais eateries tea oc oli NaF rats SerchoiS ovis oboe eee et e's ri 
Chidester, F. E.: Sarcophagid Larvae from the Painted Turtle............ 48 
Cnidosporidia of Invertebrates, Are Sarcosporidia Aberrant Forms of..... 126 
Cort, William Walter: Egg Variation in a Trematode Species............. 25 
Crawley, Howard: Note on the Stage of Piroplasma bigeminum which 
Occurs in the Cattle Tick, Margaropus annulatus.........cc0cee0eeees 87 
Crustacea, Marine, Three New. Gregarines ‘from... 5 0.60.5 06s os lees ose cee 129 
Cryptobranchus allegheniensis, Filaria cingula Parasitic in the Skin of.... 74 
Cylindrotaenia americana nov. spec. from the Cricket Frog................- 181 
Remco CSO Geer EOnd) NUGRIEAE a jar cule owe sate sie cferw Mto tl store SES ns pie bin wre ee bh oie's.8's 46 
PEC CERO LAM IG NTE SHO LAM are wiveetetetetiatere & eran eranciece: otek chara late. o:'sve, 6 ois! ole 31 sie eos 193 
Pee Aristo ith ay dS EMMALODES PECLES x wis w< siche ciliate aie ajala aie(d ose remo 20 viasine 25 
Encysted Larva of the Lung Distome, Some Notes on.................+4+ l/a 
Endamocba gingivalis (Gros) and Endamoeba histolytica Schaudinn, Fur- 
Poeee cite) (POU MOOI ALISOM! Ole ace ois aol Sinan mW Aitiawieles sare wee cose 54 
Etiology of Verruga as Deduced from a Study of the Asexual Stages of 
Banter Inia: GTM ae eR UN SS OD a odo ee eI ree ae nee ns 143 
Seymbilie. Ghes AR melevarab yas | IP ERA ee Se] Sa eee een 99 
Fantham, H. B., and Annie Porter: Significance of Certain Natural Flagel- 
lates of Insects in the Evolution of Disease in Vertebrates........... 149 
Filaria cingula Parasitic in the Skin of Cryptobranchus allegheniensis.... 74 
Flagellates of Insects, Significance of Certain, in the Evolution of Disease 
int WGI EMIES. Ms Basu db ace So etee onadod onéc )ot FORE eee OSD 149 
Foster, Winthrop D.: Two New Cases of Polyradiate Cestodes, with a 
Saimiiaty Gi te (Cases: AiredOy Si GOW pee cea cease sancti ea cces cus 7 
Further Note upon Comparison of Endamoeba gingivalis (Gros) and 
Pagamocoa. Jnstelvtica. SChauitin, o. seeps ee sts ss... essen e wen yetee 54 
Galli-Valerio, B.: Are Sarcosporidia Aberrant Forms of Cnidosporidia of 
itivextebratesiaen te cit creer ee eee erie oe ogc oe» o's am deterrents 126 
Gongylonema in the Role of a Human Parasite...............0ss eee eee ees 119 
Gongylonema scutatum, Life History of............... eee cece cece eee eeee 80 
Gregarine Parasites from Arthropoda, Some New............++++++e++0+5 27 
Gregarines, Three New, from Marine Crustacea...........----0++eeeeeees 129 


Hall, Maurice C., see Ransom, Brayton H., and Maurice C. Hall. 


206 


PAGE 
Harvard School of Tropical Medicine. Report of First Expedition to 
SVT Cie EN Or) MRENTENT )ic ok vaya 6 sick s ostine ce es acaeso ee aoe eee eee 147 
Helminthological Society of Washington, Proceedings.................. 93, 195 
Herms, William B.: Pajaroello Tick (Ornithodorus coriaceus Koch) with 
Special Reference to Life History and Biting Habits.................. 137 
lnfiusoniamebatasite an sand Pleas, New .o.o. cc. <acledcis ee eelee iene 145 
Insect) Vector of Uta, a Peruvian Disease... .... 0... <<. 2 0wcerise slneniaisien = 67 
Intermediate Hosts of the Lung Distome, P. westermanti Kerbert.......... 111 
Jewell, Minna E.: Cylindrotaenia americana nov. spec. from the Cricket 
Ere eR os wien a's sini coe ce ene deny oss as en pelea iene ee naan 181 
Katayama nosophora as a Host (note)......-. eee e eee eee e reece renee ees 50 
Krecker, Frederic H.: Filaria cingula Parasitic in the Skin of Crypto- 
branchus alleghentensis .....60.cce cece ence cece cece eee e nee enesec anes he 
Larvae, Sarcophagid, from the Painted Turtle..........-...eseeseeeeeeeee 48 
Life History of Gongylonema scutatum.... 0... cee r eee rece erence eee n ees 80 
Linton, Edwin: Cestode Cysts from Muskrat...........0+sseeees seen eeeee 46 
Macacus rhesus, Pneumonyssus foxi nov. sp., Arachnoid Parasitic in the 
Bpane Of a Monkey... 2. ccc. cient oe ee sens nie aie cintele pins dis eine wien ale 37 
Man, Parasites of: 
WirecrnOkenick Bites. On Man. .: <<. s6%s «snares see Pate See en ee eee 193 
Encysted Larva of the Lung Distome....................sceeeeeeees 175 
Further Note upon Comparison of Endamoeba gingivalis (Gros) and 
Pudamocbha iastolytica. Schaudinn.....2.5: 3.02. deen = eee 
Gongylonema in the Role of a Human Parasite................+...05- 119 
Imeem Vector of ‘Uta; a Peruvian Diseases: 0... 3.20% 55 take ee 67 
Intermediate Hosts of the Lung Distome, P. westermanit Kerbert.... 111 


Naturgeschichte der tierischen Parasiten des Menschen, 1. Teil (review) 201 
Pajaroello Tick (Ornithodorus coriaceus Koch) with Special Refer- 


enuee fo. Lite History and Biting (Habits): .... 72. 2i5.1.seeee eee 137 

Rate of Growth of the Beef Tapeworm in Human Beings (note).... 98 

Cchistosoma. gapornicum. (TOE) <0. cece osmw as Wesco Nios sienna 50 

IS CHISTOSOMMANONSONML. (TOLE)..,. cues satin ccs crete capeeeieisie Gleein tee cit eee ents 203 
Margaropus annulatus, Note on the Stage of Piroplasma bigeminum which 

iemercuaneine Cattle Lick. :.:. uirwacscm nee oon ees tes oe ineiee eee ee 87 


Mavor, J. W.: On the Occurrence of a Trypanoplasm, Probably Trypano- 
plasma borreli Laveran et Mesnil, in the Blood of the Common Sucker, | 


COLGSLOPIUSOGOMUINET SONU 0.5% ow sinisfers weteilsws © 0 wiiors xs Sb ciaeie ee sisi cents 
Metaficeys ce ect of Tick-Bites;ons Mans. . :sm6 ota mn vpn 193 
Memorins do. lustituto Oswaldo Cruz (review) = vscjcaiie » dyeupte ccm eee 148 
Musca domestica, baibernation of. (NOE) 6... ~ sos bes naives eis te eine Gee 203 
Minsicrat seesrode Cysts from..... 91: ekdhwetiee Sin ped k hee cece 46 
Newalntusotianeelarasite in Sand Wileas sews sae eee eee aie 145 
RicHEGE hE aie akin oes eos oe UDR e heny hee he RM eaaE 50, 96, 147, 201 


Occurrence of a Trypanoplasm, Probably Trypanoplasma borreli Laveran 
et Mesnil, in the Blood of the Common Sucker, Catostomus commersonu 1 
Ornithodorus coriaceus Koch, Pajaroello Tick, with Special Reference to 


ivfeiistomy.and: Biting Habits ay .<caases sein anaeewi hc abahe falR eee 137 
Pajaroello Tick (Ornithodorus coriaceus Koch) with Special Reference to 
Lite sblistomyeana Biting Habits...;. scriiilsa. evs Jarome cen eee 137 
Paragonimus westermani Kerbert, On the Intermediate Hosts of the Lung 
PUES COLIAG MIR On e's os. 0 a 0 ov .ov bau yi d4)a Fide Ue ae Senn Tig nie eee eee nn een 111 
Piroplasma bigeminum which Occurs in the Cattle Tick, Margaropus 
annulatus, more on the stage of.......4...ssas 0 daakeupatenace een 87 
Pneumonyssus foxt nov. sp., Arachnoid Parasitic in the Lung of a Monkey 
(RTCA CUS PICS Taixaivis 00's voy es ssvsin sures cana anios tent: ote 37 
Polyradiate Cestodes, Two New Cases of, with a Summary of the Cases 
PEAY Mee IAGIU EL anajeis oss va oon cs svecees ai sit Ohmnaee ce oelenin skin ann 7 
Porter, Annie, see Fantham, H. B., and Annie Porter. 
Prowazek: iErotessomevon, Sketch’ of. .i2... 0). acini ec ccd me cen ete mnne 51 
PSO OPLES CUNICUIE UTIOLE in sows nnn ca oe smccsje see ncusumdssecce tee cen eemenam 98 
Railliet. A.* gamle des) dt helaziidae. . 2... sas cpm saci ues «neue hae cree 99 


Ransom, Brayton H., and Maurice C. Hall: Life History of Gongylonema 
SCULCTLTN Rae era cienG <hik's oo oe ne ce binds pee Pete ees te ne eee 80 


PAGE 
Report of First Expedition to South America (review).................. 147 
Reviews: , 
IAT COTUGS eM rAaSteILGGmGen DACOCCINA Lr ee cores acs ccic.0iva's ves ie acs oo sud lean 148 
BAGMIGrIAS GO) INSEE IR WAIOG GETIE Son woven vedic cine hc cence nencsercss 148 
Naturgeschichte der tierischen Parasiten des Menchen, 1. Teil........ 201 
Report of First Expedition to South America. Harvard School of 
Ce eg Fear 1) eo SS SE eee rr 147 
Revision of the Genus Arhythmorhynchus, with Descriptions of Two New 
ees FhOe NONE. Fetes a eens xe vices cet cccn ccc cucnnvs 167 
RCO CVSTESHINGILC( TOLE ) a sree nays erciiade chee vio galcs sien ces edeaes 203 
Sarcocystis tenella Railliet, Some Notes and Experiments on.............. 20 
patcopeacia tarvac from the Pamiee Portle....2 5.5 .....-..0....2000n eee 48 
PeeasrosoiniE 4OpONictwms (HOLE ocas arene ras he's bc cae cacacavcnaccccssceres 50 
meemmanma wiansont (AOte hoc .c eee eee orcs ss ciak bcc Vcccncacceccsvece cco 203 
Scott, John W.: Some Notes and Experiments on Sarcocystis tenella 
SS i Ee gee Be i oh Lab ee ee 20 


Seasonal Distribution of Some Acanthocephala from Fresh-Water Hosts.. 106 

Significance of Certain Natural Flagellates of Insects in the Evolution of 
Biseaseuwa)) Vertebrates. et ave seme ere Oe Sek Sala 0 MiSs Sead owhceees 149 

Smith, Allen J., and M. T. Barrett: Further Note upon Comparison of 
Endamoeba gingivalis (Gros) and Endamoeba histolytica Schaudinn... 54 


Some New Gregarine Parasites from Arthropoda...................,.022: 27 
Some Notes and Experiments on Sarcocystis tenella Railliet.............. 20 
Some Notes on the Encysted Larva of the Lung Distome................. 175 
Strong, Richard P.: Report of First Expedition to South America. Har- 
Nava. scnoo...af Tropical Medicine: (review) .. 22... <<. loss. ccc eccccce 147 
Stunkard, Horace W.: Notes on the Trematode Genus Telorchis with 
SCRIMEMIAS OS. NEW SPECIES: <7. 5 <2 eb ee eh te MM ok cow law cas cau vusess 57 
Baead caging, cate of erowta. (nate) sa. dxird colose~s's & kaise we%si0's oc oe oo ore ole 98 
eeeworel. tite. Of Slowey | CBRE) c'ac.a~x sete oles Saw Ot bed ecy ceases 98 
Telorchis, Notes on the Trematode Genus, with Descriptions of New Species 57 
rere PARE OS rad pion Oe aye adic ecih MEI Ge ce wiesase a + & Gin scnin’nne a ceces 99 
aemee New Gregarines from Marine Crustacea......-:...2.......0..0000. 129 
Lala eS yp eel oes 2h ee ee 193 
Townsend, Charles H. T.: Insect Vector of Uta, a Peruvian Disease...... 67 
Note on the Etiology of Verruga as Deduced from a Study of the 
RSE RATA ABO elds ERAOEMCLEE sn. re Oe oe sk lc he ce saa cee ee 143 
Trematode Genus Telorchis, Notes on the, with Descriptions of New Species 57 
MeenaOne: Speirs. em Variation ih. ca. o. ee ots ee ee Poe ck 25 
Trypanoplasma borreli Laveran et Mesnil, in the Blood of the Common 
Sucker, Catostomus commersoni, On the Occurrence of............... 1 
Two New Cases of Polyradiate Cestodes, with a Summary of the Cases 
SRIRAM MUMRNORIEN IS ab aha gow, charg iicinlst dine wines Glas @ nldoiw Gamcd Mid sa aes eee cee ’ 
United Fruit Company, Medical Report (note)......................... 96, 203 
Wtaminsect, VeCtotmot a. chiiwidat DiSeaSGe.) |b ocersc sss sc loc es ccc cee cee 67 
Van Cleave, H. J.: Revision of the Genus Arhythmorhynchus, with 
Descriptions of Two New Species from North American Birds........ 167 


Seasonal Distribution of Some Acanthocephala from Fresh-Water Hosts 106 
Verruga, Note on the Etiology of, as Deduced from a Study of the 


Pa SPSE To see aM 2 cori | aU nl eee aes 0 Sr 143 
Ward, Henry B.: Gongylonema in the Role of a Human Parasite.......... 119 
Watson, Minnie E.: New Infusorian Parasite in Sand Fleas............. 145 

Some New Gregarine Parasites from Arthropoda.................... 27 

Three New Gregarines from Marine Crustacea....................00. 129 
Weidman, Fred D.: Pneumonyssus foxt nov. sp., Arachnoid Parasitic in the 

inenotea Monkey, CIMGCOEUS PHRESIES eryteet ee tac os sss cs cess ecm ete ST. 
Yoshida, Sadao: Intermediate Hosts of the Lung Distome, P. westermanii 

LESESD DET a OV GR OE Se micicl NOP NOI ARI CO IE eae eee 111 


ERRATA 


For the table on page 199, Vol. II, No. 4, substitute the following: 


Bulb about pharynx, none; ovary one (except in No. 6) 
Tail simply conoid; head rounded 
Ainpiidsmasmwideras spuatynx:..ciec.ckcicee seer ere f- communis de Man 1 
Amphids half as wide as pharynx 
Ceph, setae half long as head is wide; oes. 20%; spin. symmetrical 
-f obtusus n. sp. z 
Ceph. setae papilloid; oes. 14%; spinneret asymmetrical 
? melancholicus de Man 3 
Tail conoid, then cylindroid; head more or less truncate 
Pharynx twice long as head is wide; ceph. setae four or none 
Uterus and ovary simple; amphids minute or none 
Ovary reflexed; amph. small entering obliquely (Cylindrolaimus ?)...... 


‘f tristis Ditlevsen 4 
Ovary outstretched; no amphids, setae, or spinneret (gen. nov. ?) 
-f macrurus Daday 5 


Wtert2; ovaries rehlexed; amphid a spiral (Plectus ?)...i6.5 ncpceemsoee et 
‘f? aberrans Micoletzky 6 
Pharynx as long as head is wide 
Cephalic setae 6,3). (.2........ (Prismatolaimus ?)-f politus Daday 7 
EBACE Sciste (aa eehe's 12 tice dc Ss <bee ased eau -f brachystoma Hofmanner 8 
Bulb of pharynx distinct; ovaries 2; setae none (Ethmolaimus ?) 
? lacustris Hofmanner Os 


fHE 


Journal of Parasitology 


A QUARTERLY DEVOTED TO MEDICAL ZOOLOGY 


EDITORIAL BOARD 


FRANKLIN D. BARKER ALLEN J. SMITH 

The University of Nebraska The University of Pennsylvania 
CHARLES F. CRAIG JOHN W. SCOTT 

Medical Corps, U. S. Army The University of Wyoming 
WILLIAM B. HERMS CHARLES W. STILES 

The University of California U. S. Public Health Service 
BRAYTON H. RANSOM RICHARD P. STRONG 

U. S. Bureau of Animal Industry Harvard Unicersity 
WILLIAM A. RILEY JOHN L. TODD 

Cornell University McGill University 


ROBERT T. YOUNG 
The University of North Dakota 


VOLUME. 3 
1917 


Managing Editor 
HENRY B. WARD 


The University of Illinois 
URBANA 


FOREIGN COLLABORATORS 


B. BLACKLOCK - School of Tropical Med., Univ. of Liverpool, England 
RAPHAEL BLANCHARD - - - College of Medicine, Paris, France 
MAX BRAUN - - - - - University of Konigsberg, Germany 
ANTON BREINL, Australian Institute of Tropical Medicine, N. Queensland 
J. BURTON CLELAND - -_ Bureau of Microbiology, Sydney, Australia 
H. B. FANTHAM - - - School of Tropical Medicine, Liverpool 
O. FUHRMANN - - - - University of Neuchatel, Switzerland 
B. GALLI-VALERIO - _ Laboratory of Parasitology, Lausanne, Switzerland 
L. GEDOELST - - > State Veterinary School, Brussels, Belgium 
L. A. VON JAGERSKIOLD - - Zoological Museum, Goteborg, Sweden 
T. HARVEY JOHNSTON =- . - University of Queensland, Australia 
MARIE V. LEBOUR - - - - - University of Leeds, England 
Reel PIPER y= - - . - School of Tropical Medicine, London 
A. LOOSS - - - - - - - - - - Leipzig, Germany 
P. S. DE MAGALHAES - - Medical Faculty, Rio de Janeiro, Brazil 
AL. MRAZEK - -  - += = + Bohemian University, Prague 
WILLIAM NICOLL, Australian Institute of Tropical Medicine, N. Queensland 
1,,ODHNER = - -+- - £Zoolopical Museum, Christiania, Norway 
E. PERRONCITO - - . - - - University of Turin, Italy 
TH. PINTNER - . - - - - University of Vienna, Austria 
AS RAILEIED. = - - - National Veterinary School, Paris, France 
S. VON RATZ - - - - Hungarian Veterinary Academy, Budapest 
A. E. SHIPLEY - - - - Christ’s College, Cambridge, England 
T. SOUTHWELL - - - - Director of Fisheries, Calcutta, India 
GEORGINA SWEET - - - - University of Melbourne, Australia 
S. O. YOSHIDA - - - - - Osaka Medical Academy, Japan 


F. ZSCHOKKE - - - = Zoological Institute, Basel, Switzerland 


CONTENTS OF VOLUME III 


SEPTEMBER, 1916. NUMBER 1 pice 


CONTRIBUTIONS TO THE Stupy oF Parasitic Protozoa. III. Nores on 
MyxosporipIA Founp IN SoME FRESH-WATER FISHES OF JAPAN, WITH 
THE DESCRIPTION OF THREE NEW SPECIES. ROKUSABURO KuDo........ 3 

(With four text figures) 


Notes oN Two Free-Livinc LArvAL TREMATODES FROM NortH AMERICA. 
DsliDya here gop WS Nas, Beh eek scree oem ERC EET EP Pe en 10 
(With one plate) 


ON THE ANATOMY AND RELATIONSHIPS OF SOME NorTH AMERICAN TREMA- 
TODESSas GLORAGED INVig: SUN ROAR a pet eee teraoee ed cycys ero. e dalee ck Son or adiveion 21 


DAUERCYSTFORMATION OF Trichomonas intestinalis. KENNETH M. Lyncu.. 28 
(With two text figures) 


Notes oN Two CESTODES FROM THE Spottep StTING-Ray. Epwin Linton.. 34 
(With one plate and two text figures) 
A CASE OF THE OCCURRENCE OF Ascaris triquetra SCHRANK IN Docs. 
JBL, {GL AUNTS 8 at ete aid aA Gee Gee ctor & os ane a 39 


(With six text figures) 


JPQCNa TS: UNNID) DCIS SS a aA Dela eee Met BT oe Ae ee 42 


. 


DECEMBER, 1916. NUMBER 2 


THe Errects oF RADIATION ON THE DEVELOPMENT oF Trichinella spiralis, 
witH Respect To Its APPLICATION TO THE TREATMENT OF OTHER PARA- 
Sipiee DiseAces. FE. “Tyzzer AND JAMES A. HoNEIy..............% 43 

(With one plate) 


Notes oN SoME NEMATODES FROM FRESH-WaATER FisHES. HENRY B. Warp 
LATOR DNS, AB SIN UNO NE ee O85 SS en BR GEAR Oe Gao Se eee eae 57 


(With one plate) 


OBSERVATIONS ON Potycystip GREGARINES FROM ARTHROPODA. MINNIE E. 
VINOATISTOIN T= Brera a en Re acicietlo ks tiara Hho re Bre cee rr 65 
(With one plate) 


On A TREMATODE Larva ENcysTED IN A Cras, Helice tridens (pE HAAN). 
SCAR a OS EIT EVA Cty sre eye Sieg ee Mery Sere RO er icici sc alee o0ea os a stcldietns AG: 
(With two text figures) 


Cytoleichus peurosci, A NEw ARACHNoID PARASITE FouND IN THE DISEASED 
LunGs oF A Prairie Doc, Cynomys ludovicianus. Frep D. WEmMAN 82 
(With two plates) 


Book Review ANnp NOTE.:..:..........-. me ARR pe Ne has Sake 3 2 ee 90 


iv | CONTENTS OF VOLUME III 


MARCH, 1917. NUMBER 3 PAGE 


On THE SPoROZzOON PARASITES OF THE FISHES OF Woops HOLE AND VICINITY. 
I. FurtHER OBSERVATIONS ON Myxobolus musculi FRoM FUNDULUS. 
GLUON a ihweanp, 5: eet) |) A Be ei I oye 8 55) eS St iy 9.47 91 
(With three text figures) 


NoTES ON THE CERCARIAE OF THE BitTER Root VALLEY, MonTANA. ERNEST 
MR OROUT) SANIST cc se hace ce ecg ia eee Pate oe tree merce eee 105 
(With one plate) 
Tue DEVELOPMENT OF GREGARINES AND THEIR RELATION TO THE Host Tis- 
suE: (1) In Stenophora lactaria Watson. MINNIE Watson Kamm 124 
(With two plates) 


TE MOERCARTARVORUNATAL. | E.cG. (GAWSTONicis 2 chresioepae eon sororities sm A 


Note oN A SpeEcIES oF NoseEMA INFECTING Attacus cynthia Drury. 
SinCo wun) SEIN G0 ne Pear Arn resi San oc aod God ae c 136 
(With eight text figures) 


Notes on Porocephalus globicephalus. Turstt T. Jos ANp A. R. Cooper.. 138 
1ELoayre [PSG aa Ae ee nan indni icign co Sc.6.50 dactogattadscoo 139 


TN OTS ews, Glp ORs Crone EC EE Se Cri erte  e dnoneaooGe es 141 


JUNE, 1917. NUMBER 4 


Endamoeba buccalis. 1. Irs MULTIPLICATION AND PEeErRtopiciIty. NADINE 
ICOM QACICN, Ql ne meric A SE eS Pee Aol dlodance - 143 
' (With one text figure) 


ON THE SpoROZOON PARASITES OF THE FISHES OF Woops HOLE AND VICINITY. 
II. AppITIONAL OBSERVATIONS ON Myxobolus musculi oF FUNDULUS AND 
A Nearty Revatep Species, M. pleuronectidae or Pseudopleuronectes 
GHLETCONUS ANG. W.. LLATIN 5.0's::.0:) oh oop Re eee cee nce eee 150 
(With one plate) 


CONTRIBUTIONS TO THE Stupy or Parasitic Protozoa. Il. Myxrobolus 
toyamai Nov. SPEC., A NEw MyxosporipIAN ParAsITE IN Cyprinus 
CakPiO vic) SROKUSABURO KUuDO.. ... «525/25 Seer eek ore ea ees it ee 163 

(With two plates) 


THE OccurrENCE OF Bothriocephalus liguloides LeucKART, WITH ESPECIAL 
REFERENCE TO ITs DEVELOPMENT. SADAO YOSHIDA.............e+ceeee 171 
(With one text figure) 


A FurtHer NOTE ON THE Lire-History or Gongylonema scutatum. BRAYTON 
Hi RANSOM AND Maurice C. HALL... .55)62.2 005 deene ee 177 


NOTES | caereeRnNe aoe esis a ce cea oc wen oe be ee eR eo 182 


The numbers of Volume III of the JourNat or PARAsIToLoGy were mailed as 
follows: ; 


No. 1. Nov. 22, 1916. No. 3. May 1, 1917. 
No. 2. Feb. 27, 1917. No. 4. July 10, 1917. 


The 
Journal of Parasitology 


A Quarterly Devoted to Medical Zoology 


Volume III 


EDITORIAL BOARD 


FRANKLIN D. BARKER ALLEN J. SMITH 

The University of Nebraska The University of Pennsylvania 
CHARLES F, CRAIG JOHN W. SCOTT 

Medical Corps, U. S. Army The University of Wyoming 
WILLIAM B. HERMS CHARLES W. STILES 

The University of California U. S. Public Health Service 
BRAYTON H. RANSOM RICHARD P. STRONG 

U. S. Bureau of Animal Industry Harvard Unicersity 
WILLIAM A. RILEY JOHN L. TODD 

Cornell University McGill University 


ROBERT T. YOUNG 
The University of North Dakota 


HENRY B. WARD, MANAGING EDITOR 
The University of Illinois 
URBANA 


Lhe Joupnials0i t.arasitology 


Volume 3 SEPTEMBER, 1916 Number 1 


CONTRIBUTIONS TO THE STUDY OF PARASITIC 
PROEOZO. IIT. 


NOTES ON MYXOSPORIDIA FOUND IN SOME FRESH-WATER FISHES OF 
JAPAN, WITH THE DESCRIPTION OF THREE NEW SPECIES 
(WITH FOUR TEXT FIGURES) 


ROKUSABURO Kubo 


THE IMPERIAL SERICULTURAL EXPERIMENT STATION, NAKANO, TOKYO 


In a former paper (1915) I described from a morphological as well 
as developmental point of view, a new species (My-vobolus toyamai 
Kudo) from the branchial lamellae of a carp. The present paper is 
the result of study upon Myxosporidia found since that time. I am 
now working on the life-histories of the new species described below 
with the hope of reporting them later. 


1. Myxosoma dujardini Thel. 

Eight cysts (the largest being 200 in diameter) of round shape, 
were found in the branchial lamellae of a carp 23 cm. in length. The 
seat of the parasite was, as in Myxobolus toyamai, the connective tissue 
of the gill-filament. 

The results of observations upon the spore coincide for the most 
part with the description of Thélohan (1895). I wish, however, to 
give here details about the polar capsule and the polar filament, as 
Thélohan failed to mention them: length and breadth of the polar 
capsule 6-7 and about 2, respectively, and the length of the polar 
filament about 70. 


2. Zschokkella acheilognathi n. sp. 

Vegetative form. Large ones are generally visible to the naked 
eye as small, opaque, more or less regular, usually subspherical masses, 
occupying various parts of the gallbladder and especially of the gall- 
duct (Fig. 1). The size varies with age up to a maximum length of 
720 by a breadth of 550p, and the thickness of one individual is about 
uniform throughout, but in many specimens it differs from 5 to 30p 
according to the size of the myxosporidium. Their bodies are very 
flexible and easily doubled up, representing, in sections, various forms. 
The vegetative stage in sections resembles much that of Sphaeromyxa 


4 THE JOURNAL OF PARASITOLOGY 


hellandi, observed by Auerbach (1912), both in form and structure. 
The body is colorless in both young and old. In its fresh condition 
the protoplasm can be seen to be clearly differentiated into finely 
granulated reticular ectoplasm and greatly vacuolated endoplasm. In 
the younger form (15 to 30u in greatest diameter) lobose pseudopodia 
are well developed. The myxosporidium moves about more or less 
actively in the bile by the constant emission of pseudopodia. No clear 
evidence of the active emission of pseudopodia exists in older individ- 
uals. The ectoplasm of some more advanced specimens shows in sec- 
tion two structures; the outer layer, comparatively thin but uniformly 
about 2y in thickness, presents very fine striations, while the remaining 
part is finely alveolated, having an average thickness of 6 to Su. 


Stem = 
=~ = Fa eS D 
EEE 
s = nn 


ER es ies SS 


Ry 
Yk 


a b. 
Fig. 1—Zschokkella acheilognathi n. sp. a, Gall-bladder of Acheilognathus 
with many myxosporidia, X10; b, oblique cross section of the infected gall- 
duct, < 200; c, part of the cross section of the parasite, showing the differ- 
entiation of the protoplasm, < 1,000; d, to g, spores: in g, filaments are extruded, 


< 1,500; h, stained spore, about X 2,250; k, young myxosporidium,  X 1,000. 


Auerbach (1910b) seems to have observed similar structure in the 
outer layer of the ectoplasm in Sphaeromyxa hellandi, sketching a sur- 
face view of an individual fixed in formol. It takes stains much 
more deeply than the endoplasm, so that in sections it shows clear 
differentiation of the protoplasm much better than was shown in 
My-xobolus toyamai. The endoplasm contains vegetative nuclei as well 
as generative nuclei in several stages of spore formation. It is poly- 
sporous, according to the observation made up to the present time. 
In this regard, it is quite different from Zschokkella hildae, 11 which, 
after Auerbach (1910a), single and double spore formation occurs. 

Spore. Generally oval with round poles, very often more or less 


KUDO—MYXOSPORIDIA FROM JAPAN 5 


hemispherical, somewhat attenuated symmetrically at both ends of the 
flat side of the spore. Several modifications in form and size, however, 
are also found in the present case as in other forms. It is usually 
10 to 14 long by 6 to 7 wide. The thickness is almost equal to the 
width. Shell bivalve; the line of junction being oblique to the longi- 
tudinal axis of the spore. Parallel to the line of junction, fine stria- 
tions run longitudinally on the spore coat. A polar capsule, round 
in shape, with a diameter of 2 to 3, at each end of the spore, takes 
stains very deeply. Polar filaments were easily extruded by the appli- 
cation either of mechanical pressure or KOH-solution, and are well 
stained after my method (1913). The fully extruded polar filaments 
were 65 to 70p long. 

Habitat. This species is found quite abundantly in the gallbladder 
of Acheilognathus lanceolatum Temm. et Schl., commonly found in 
brooks in the vicinity of Tokio. Out of twenty-four fish (8 to 12 cm. 
long) examined in May, 1915, twenty-one were found to have har- 
bored the parasite; thus the rate of the infection rises above 80 per 
cent. Matured, large vegetative forms were very often found in great 
numbers in the gallduct, while in the gallbladder of the same host I 
could find only a small number of isolated spores. Klokacewa (1914) 
described a somewhat similar form of spore from Carassius vulgaris, 
without finding the vegetative form. The species in question, though 
its vegetative form differs apparently from that of Zschokkella hildae 
seems to belong to that genus. Up to the present time two species of 
the genus have been reported, that is, Zsch. hildae and Zsch. nova, 
since Auerbach (1910) created it. The former apparently differs from 
the present form in several points. Now the dimensions of the spore 
of the latter seem to correspond very nearly to the myxosporidium in 
question. As it lacks, however, all other details, it is impossible to 
make accurate identification, so I treat this species as a new one, calling 
it Zschokkella acheilognathi. 


3. Myxobolus fuhrmanni Auer 


Isolated spores of this form occur very often in the bile of the 
loach (Misgurnus anguillicaudatus). The vegetative form has not yet 
been found by me. The description of the spore by Auerbach (1909) 
coincides well with my observations, except that I found the thickness 
of the spore coat to be uniform, whereas Auerbach’s observation was 
in effect that the shell is especially thick at the posterior end of the 
spore. I wish to mention that the length of the polar filament in the 
present case is 100u. Nearly 50 per cent. of the said fish, examined in 
September, 1915, were infected by this Myxobolus. In all cases, how- 
ever, the infection seems to be carried to a very slight degree. 


6 THE JOURNAL OF PARASITOLOGY 


4. Myxidium sp. 

This form, together with the following two new species, are also 
found in the gallbladder of the loach. The vegetative form has not 
yet been observed. Two per cent. of the fish studied in September, 
1915, were infected. The spores are mostly found separated from each 
other, floating in the bile. One side of the spore is, in most cases, 
more convex than the other. The sporoplasm usually occupies the 
whole inner space of the spore, except the polar capsules, and shows 
fine granulations in the natural condition as in the case of Myxidium 
giardi or Myxobolus pfeifferi, and also shows a fine alveolar structure 
in stained preparations. It contains two nuclei of almost equal size. 
Shell bivalve, the line of junction of which is straight. On the surface 
of the shell, fine striations run longitudinally parallel to the line of 
junction. The dimensions are: length, 15 to 18», breadth 6 to 7p, 
length of polar capsule 7 to 8u, and that of polar filament 60 to 70x. 


Fig, 2—My-xidium sp. a, stained spore, X 1,750; b, spore with extruded polar 
filaments, > 1,000. 


Ishii (1915) recently described a new species, Myxidium anguillae, 
from an eel. The form in question apparently differs from any of the 
species reported up to the present time. I hope to identify the species 
after studying it more closely. 


5. Chloromyxum misgurni n. sp. 

Vegetative form. Mostly round, often of irregular form. From 
a side view it assumes a semicircular shape. From the more or less 
flat surface many fine root-like pseudopodia extend. They are more 
clearly visible in younger specimens, where the spore formation has not 
yet begun. There is no clear differentiation of protoplasm. It is finely 
vacuolated on the whole. With the pseudopodia, the myxosporidium 
probably creeps along the surface of the epithelial layer of the gall- 
bladder, so that many individuals in different stages of development 
are found in section preparations, closely attached to the epithelial 
cells with their peculiar pseudopodia. The size varies with age, the 
largest being 50 in greatest diameter, with the maximum thickness of 
20. Individuals with six to eight spores are of common occurrence ; 
those with twelve to sixteen spores, however, occur rarely, and it is 
very seldom that only two spores are found in one myxosporidium. 

Spore. Spherial, slightly attenuated at the anterior end. Shell 


KUDO—MYXOSPORIDIA FROM JAPAN 7 


bivalve; the line of junctiom straight. Parallel to the ridge which 
marks the line of junction very clearly, run fine longitudinal stria- 
tions. Four polar capsules are situated in the anterior end. In the 
finely granulated sporoplasm two nuclei of equal size are found. The 
dimensions of the spore are: length, 8 to 9u, breadth 6 to 7p, thickness 
5 to 6, length of the polar capsule 2 to 3u and that of polar filament 
28 to 35y. 

Habitat. In the gallbladder of Misgurnus anguillicaudatus Cantor. 

Of the fish examined in September, 1915, 73 per cent. were found 
to be infected. ; 

Of forms known up to the present time, Ch. fluviatile (Thélohan, 
1895) seems to be nearest in size and form of the spore and seat of 
infection to the Chloromyxum mentioned. The form and size of the 


Fig. 3—Chloromyxum misgurni n. sp. a, front view; b-c, side view of the 
spore in natural condition, 1,750; d, stained spore, < 2,625; e, spore with the 
extruded polar filaments, < 1,750; f, g, two young myxosporidia in section, 
s< 1750. 


pseudopodia and the structure of the spore of the type studied differ 
from that described by Thélohan. Therefore, I propose to name this 
Chloromyxum misgurnt. 


6. Chloromyxum fujitai n. sp. 


Vegetative form. Mostly round, sometimes irregular. There is 
no clear differentiation of the protoplasm. The endoplasm is highly 
vacuolated, the ectoplasm heing hardly visible. The largest one was 
4Quin diameter. They float about in the bile, so that in sections of 
infected gallbladder they are found in the gall apart from the epi- 
thelial layer, by which fact we can easily distinguish them from 
Chloromyxum anguillicaudati, even when both forms occur in the 
same gallbladder. Disporous and polysporous, with up to eight spores. 

Spore. Circular in general; often attenuated at the anterior end 
Shell bivalve; the line of junction not being straight, but very thick. 


8 THE JOURNAL OF PARASITOLOGY 


The shell has peculiar thick ridges running longitudinally on the sur- 
face. Near the anterior end of the spore two small circular markings 
are clearly visible, in preparations well stained with Heidenhain’s iron 
hematoxylin, one on either side of the line of junction, from which the 
markings recede on both valves. These two circular markings are not 
the exits for polar filaments, because 1t is clearly shown in prepara- 
tions stained with Giemsa’s solution that the four polar capsules 
have their independent exits. The form of the spore takes different 
aspects by the presence of the characteristic markings resembiing par- 
tially those of Hoferia cyprini (Doflein, 1898) and Chloromyxum kot 
(Fujita, 1913). In optical cross-section, the spore represents an out- 
line quite like a cog-wheel with twenty to twenty-two ridges, including 
the widest ridges, which mark the line of junction of the shell. The 


Fig. 4—Chloromyxum fujitai n. sp. a, young vegetative form, X 1,750; 
b, c, anterior (b) and posterior (c) view of a stained spore, X 1,750; d, optical 
cross section of the.same spore as the above, x 1,750; e, f, side views of stained 
spores, < 1,750; g, side view of stained (Giemsa) spore, X 1,750. 


thickness of the ridges varies regularly. The thickest ones are located 
where a plane perpendicular to that of junction cuts the shell longi- 
tudinally, others decreasing in thickness as they approach the line of 
junction. Four polar capsules occupy the anterior half of the spore. 
The sporoplasm contains two nuclei of almost equal size. The 
dimensions are: length 10 to 12, breadth 8 to 10, length of polar 
capsule 2 to 3y, and that of the polar filament 23 to 30x. 

Habitat. In the contents of the gallbladder of Misgurnus anguilli- 
caudatus. The occurrence is much rarer than that of the former one, 
showing about 5 per cent. in September, 1915. 

Of all descriptions from this genus that of Fujita (1913) alone 
described similar markings of the spore of Chloromyxum koi from 
the gallbladder of the carp. However, we find a great difference 


KUDO—MYXOSPORIDIA FROM JAPAN 9 


in form and in the number of ridges. There is also a great difference 
between the size and structure of the spore, and in the number of the 
spores found in a vegetative form. So I think this species is a new one. 
In honor of Dr. T. Fujita, who was the first to study myxosporidia in 
Japan and to discover the spore of this type, I give the name 
Chloromyxum fujitai. 

Multiple infection of the above-mentioned four species takes place 
very often. 

Concerning the pathological effects, I have but little to report. No 
visible external change could be noticed in any of the infected fishes. 
But, as in the case of Acheilognathus, we found often that the gall- 
duct had been filled up with a great number of the Zschokkella 
(Fig. 1), therefore it is certain that the secretion of the bile into the 
duodenum must be greatly disturbed. Such is the case with the highly 
infected loach, the gallbladder of which has an opaque appearance. It 
is, however, difficult at present to state the real effects of the parasite 
upon the host. 

REFERENCES CITED 


Auerbach, M. 1909. Bemerkungen tiber Myxosporidien. Zool. Auz., 34: 65-82. 
1910. Biologische und Morphologische Bemerkungen tber Myxosporidien. 
Zool. Anz. 35:57-63. 1910a. Die Sporenbildung von Zschokkella und das 
System der Myxosporidien. Zool. Anz., 35: 240-56. 1910b. Cnidosporidien- 
studien. Zool. Anz., 35: 767-77. 1912. Studien iiber die Myxosporidien der 
Norwegischen Seefische und ihre Verbreitung. Zool. Jahrb., Anat., 34: 1-50. 

Doflein, F. 1898. Studien zur Naturgeschichte der Protozoen. III. Ueber 
Myxosporidien. Zool. Jahrb., Anat., 11: 281-350. 

Fujita, T. 1913. On a new species of Chloromyxum from the gall-bladder 
of the carp. Annot. Zool. Japan., 8: 257-59. 

Ishii, S. 1915. Myxosporidiosis of the eel. (Japanese.) Trans. Tokyo 
Zool. Soc., 27 : 372-82. 

Klokacewa, S. 1914. Ueber die Myxosporidien der Karausche. Zool. Anz., 
44: 182-86. 

Kudo, R. 1913. Eine neue Methode die Sporen von Nosema bombycis 
Nageli mit ihren ausgeschnellten Polfaden dauerhaft zu praparieren und deren 
Lange genauer zu bestimmen. ZXool. Anz., 41: 368-71. 1915. Contributions to 
the Study of Parasitic Protozoa. II. Myxobolus toyamai n. sp., a new 
myxosporidian Parasite in Cyprinus carpio L. (Japanese.) Trans. Tokyo Zool. 
Soci 2 golgeaae 

Thélohan, P. 1895. Recherches sur les Myxosporidies. Bull. scient. France 
et Belg., 26: 100-394. 


NOTES ON TWO . FREE-LIVING LARVAL TREMA- 
Toots FROM NORTH AMERICA 


Henry B. Warp 


The life history of parasitic worms has always been a subject of 
especial interest and no part of it commands more careful attention 
than that which deals with the brief stages of free existence, for here 
is the point at which the organism effects its transfer from one host 
to another. It is only in the few highly specialized types that the para- 
sitic habit endures in unbroken succession from host to host and the 
parasite is transferred by the agency of the organism in which it is 
living. This is the case with the Plasmodium malariae and all other 
parasites transferred by a blood sucking host to a new environment in 
which usually if not always a new generation is developed. It is also 
the case with the trichina and other encysted parasites which are 
acquired by a new host thru its carnivorous habit and which develop 
into a new form or stage of the life history in this new host rather 
than as a new generation. 

In the simpler cases, however, the parasite abandons at intervals 
the parasitic mode of life and adopts a free-living habit for a stage 
of its existence or for a generation that alternates with the parasitic 
type. The free-living generation or stage is of especial interest because 
it affords the opportunity for the infection of a new host and also 
furnishes a point of attack in the life cycle which is vulnerable so 
that the readjustment of environmental conditions may block the 
transfer and prevent the infection. Such a readjustment may result 
from the natural operation of external forces, as when a very dry 
season eliminates the small ponds or swamps in which the free stage 
develops and thru which it secures means of transfer to the new 
host. Or the change may be brought about by the introduction of 
hygienic regulations that are drafted to prevent the parasite from 
reaching its new host, as the installation of a new sewage system may 
divert the human feces with tapeworm eggs from the lake into which 
they were formerly discharged and thus prevent infection of the fish 
host in that lake with the bladder stage of the fish tapeworm (Diboth- 
riocephalus latus) ; consequently the spread of the tapeworm in the 
human host is checked. 


* Contributions from the Zoological Laboratory of the University of Illinois, 
No. 71. 


WARD—FREE-LIVING LARVAL TREMATODES 11 


Attention has been forcibly directed in the past year to the free- 
living stages of trematodes by the work of Japanese and European 
investigators on the life history of the Lungfluke (Paragonimus) and 
the Bloodfluke (Schistosoma). As a result of these studies it is possi- 
ble now for the first time to draw a reasonable picture in outline of 
the development of these forms. Almost nothing has been ascer- 
tained concerning the free-living stages of such parasites in North 
America. A few observations were made years ago by Leidy and 
recently Cort has published a careful study of some larval trematodes ; 
but together these cover only a few of the many North American 
species of fluke. The importance of placing on record all data leads 
me to print here observations made some years ago although they are 
yet unfortunately incomplete. For the satisfactory interpretation of 
these observations a preliminary statement may be made here regarding 
the process of development as found in the trematode. 

Two free-living stages recur in the development of most flukes. 
From the egg develops a small ciliated larva, designated a miracidium 
by Braun, which is evidently dependent upon water for its distribution. 
It remains within the shell until it has reached its full development ; 
thereafter contact with the water is sufficient to open the shell and 
bring about the escape of the embryo. Active migration through the 
water permits it to reach and infect the secondary host, a mollusk. It 
is a somewhat striking fact that in spite of the constant and abundant 
production of eggs and embryos, no records that I have found note the 
occurrence of such embryos in plankton or other fresh-water collec- 
tions. The absence of records can not be attributed to the small size 
of the miracidia since other even more minute objects fall constantly 
within the ken of the microscopist engaged in the study of fresh-water 
organisms, and some that are apparently very difficult to detect have 
been studied and described in detail. It may be due to the extreme 
delicacy of the larvae which are thereby readily subject to accidental 
destruction. Certainly they go to pieces almost as soon as they are 
collected. 

The second free-living stage in the life history of the trematode 
comes when the cycle of development within the mollusk is completed 
and the transfer to the adult host takes place. This transfer occurs in 
the cercaria stage and of course may be direct if the mollusk is eaten 
by a suitable host. Yet one may safely infer that this is not the usual 
method since most cercariae are so well adapted to a free aquatic 
existence. The ordinary cercaria possesses a well developed swimming 
organ in the tail which characterizes this stage and is cast off when the 
larva reaches a new host or a place of encystment, as the case may be. 


12 THE JOURNAL OF PARASITOLOGY 


This swimming tail is reduced in a few types and absent only very 
infrequently. 

In other cases the tail is not only present but powerful and displays 
various modifications, such as bristles, folds, branches, lateral mem- 
branes, etc., that increase its functional value. No one who watches 
living cercariae in the laboratory under experimental conditions can 
doubt that they are robust swimmers and naturally depend on that 
method for their transfer from the mollusk to the final host. When 
infected snails are kept in an aquarium jar on the laboratory tabie, 
the cercariae swarm out voluntarily at certain times in great numbers 
and in many cases can be seen with the unaided eye swimming actively 
about in the water. They do not confine themselves to the sides or 
bottom of the vessel but seem to seek equally the open water and in 
general to conduct themselves under such circumstances like other 
plankton organisms: protozoa, rotifers, and entomostraca in the same 
aquarium. 

These cercariae are large objects among the microscopic aquatic 
organisms; they are produced in great abundance and infected mol- 
lusks are also abundant and widely distributed. And yet there are 
almost no records of the occurrence of cercariae in the voluminous 
reports on fresh water plankton and aquatic life. I am at a loss myself 
to explain this condition. [ have seen them many times in fresh water 
collections, but only when the material was examined very soon after 
it was taken in the net. Usually the number of specimens secured 
was too limited to permit of satisfactorily determining the structure 
and relationships of the form. In two cases, however, the cercaria was 
so peculiar as to justify this record of its occurrence even though the 
description is incomplete in certain respects. 


CERCARIA ANCHOROIDES HO0v. Spec. 


The first species to which attention is called was abundant at Lake 
St. Clair in 1893 and was described very briefly in a preliminary report 
(Ward, 1894), as follows: 

In the tow was found but one helminth, a form which challenged 
attention the first of our stay. It is a free-swimming Cercaria, closely 
allied to C. mirabilis Braun, having a prominent tail terminating in two 
flat blades at right angles to the main body. The distome is enclosed 
within the tail of the Cercaria which has then more or less the appear- 
ance of an anchor with wide flukes. From one to four of these were 
taken from both top and bottom tow every day from July 27 to August 
5 when it suddenly ceased to be found. Efforts to find the intermediate 
and primary host were alike unsuccessful. This form differs consid- 


WARD—FREE-LIVING LARVAL TREMATODES 13 


erably in size from those deseribed by R. R. Wright and M. Braun and 
is probably a new species. 

The cercaria attracted attention by its very active movements, 
which were so peculiar that it could be readily picked out from other 
material in a glass dish. While found in collections from the surface 
as well as in those taken by a runner net and representing thus the 
fauna near the bottom, yet it was three times as frequent in the latter 
as in the former. This probably indicates that the cercariae were dis- 
charged from snails or other mollusks living on the bottom in the 
region where the collections were made. 


Viewed under a lens the organism appeared as a minute object 
shaped like a hammer or anchor and swam through the water by a 
succession of violent jerks. The body moved with the flukes of the 
anchor in advance and propelled itself by throwing the flukes alter- 
nately right and left. In this movement the flexure occurred in the 
stem of the anchor about one third of the distance from the flukes to 
the head, and the blades did not move separately but maintained con- 
stantly the same relation to each other and to the adjacent part of the 
stem. The motion recalled distinctly the sweep of a double headed 
paddle as it is passed from the one side of a canoe to the other. 

A more careful examination showed that the stem of the anchor 
was flat and also the flukes which extended from it nearly at right 
angles but were curved a little near the outer end. The head of the 
anchor appeared, however, nearly round, being enlarged and enclosing 
a small object which lay in a clear, fluid-filled chamber at the extreme 
head end of the stem (Fig. 2). 

The stem of the anchor measured about 2 mm. in length and the 
flukes were each 0.53 to 0.6 mm. long, though on account of the curve 
their tips were only 0.84 mm. apart. The flukes varied in width from 
0.24 to 0.34 mm. and the stem was 0.28 mm. in maximum width but 
was reduced to 0.2 mm. at the region of transition from the flat base 
to the rounded head. At its widest part near the outer end this region 
had increased again to about 0.3 mm. in diameter. 

Under the microscope one could readily distinguish that the object 
in the enlarged end was a small distome. It lay with the oral sucker 
near the apex of the chamber and with the opposite end turned towards 
the flukes of the anchor. In no case was it coiled, twisted, or crumpled 
together in life but lay flat and straight with abundant space in the 
chamber for it to be extended to full length. When the distome lay 
flat on the slide, the base of the stem and the flukes of the anchor stood 
on edge, thus the breadth of the distome was at right angles to the flat 
surface of the stem and flukes. 


14 THE JOURNAL OF PARASITOLOGY 


The living distome was faint sulphur yellow with a reddish brown 
intestine; the anchor stem and flukes were dark by transmitted light 
and white or faint yellow by reflected light. 

Under the pressure of the compressor the young distome (Fig. 1) 
was forced out of the sac in which it was contained. It emerged at the 
extreme tip where there seemed to be a preformed opening and left 
wrinkled and collapsed a thin walled sac in which it had been enclosed. 
The base of the stem and the flukes of the anchor remained entirely 
unchanged and often moved about actively in the water for some time 
after the distome had escaped, beating alternately right and left in the 
same manner as before and moving freely through the open water 
just like those specimens in which the distome was still enclosed in 
the sac. 

The young distome, freed from the sac in the tail, as the anchor 
should properly be called, measured in life 0.64 mm. in length by 0.288 
mm. in breadth. The oral sucker was sub-terminal or ventral, being 
separated 0.016 mm. from the extreme anterior tip; its diameter was 
0.16 mm. and its orifice measured 0.048 0.064 mm. The ventral 
sucker though conspicuous was a little smaller than the oral, measur- 
ing 0.128 by 0.144 mm. and being separated from the anterior tip by 
a distance of 0.27 mm.; its orifice measured 0.04 by 0.072 mm. The 
pharynx was 0.064 mm. in diameter, and in some cases even a little 
longer. 

The intestine was large, broad, wavy in outline, and filled with a 
dark reddish brown fluid containing numerous highly refractive gran- 
ules. The main duct of the excretory system extended from the poste- 
rior tip to the acetabulum and two longitudinal trunks were conspicu- 
ous on the right and left sides of the worm, outside the intestinal crura. 
They were not straight but much twisted or thrown into short heavy 
wavy loops from which fine branches extended towards the margin 
and gave rise to still finer branches. In one specimen a transverse con- 
nection was demonstrated just anterior to the acetabulum extending 
from the median posterior trunk to the left longitudinal trunk. It was 
also noticed that the right canal in the same worm was certainly larger 
near the center of the body than near either end. This condition would 
indicate that a connection existed at the center in this tube also. One 
would not go far astray in interpreting the median posterior stem as 
the bladder from the apex of which near the acetabulum branches 
extended right and left to divide again near the margin on each side 
of the body into anterior and posterior trunks. 

On the ventral surface of the body appeared a transverse slit just 
15 in front of the anterior margin of the acetabulum in the living 
specimen, or halfway between it and the oral sucker in a contracted 
alcoholic specimen. While the ducts connected with it were not yet 


WARD—FREE-LIVING LARVAL TREMATODES 15 


developed or at least demonstrable there seems little doubt that this 
represents the genital pore. In the center of the body behind the ace- 
tabulum and between the intestinal crura three faint masses were dis- 
cernible. The two smaller bodies near each other slightly oblique to 
the axis and furthest posteriad, are probably the testes and a single 
larger mass between the testes and the acetabulum is no doubt the 
ovary. These structures were faint, especially the ovary, and the 
connecting ducts of the genital system were not yet.apparent. The 
three bodies differ slightly in size and location in different specimens. 

The sac at the head of the anchor stem had on the outer surface 
peculiar vacuolated wart-like protuberances. These may have been 
sensory structures. In the outer wall were also two sets of fibers— 
probably muscular—of which those in the transverse series are closer 
together and very regular whereas the longitudinal fibers are neither 
so abundant nor so regular. The wall of the sac adjacent to the cavity 
consists of a thin epithelial layer. 

In 1885 R. Ramsey Wright found a single specimen of a similar 
form swimming actively in a fresh water aquarium at Toronto. He 
published a brief note (Wright, 1885) in which the form was inter- 
preted as a free-swimming sporocyst. Leuckart to whom the specimen 
was sent with notes and a sketch published (1886) a more extended 
description with a copy of Wright’s drawing. From this account it 
is easy to determine that the form is very similar to that described 
above but not identical with it. Both the length which is “nearly 
1 mm.” (Leuckart) as against 2 mm. in the new species and the form 
of the tail as well as of the young distome indicate the specific dif- 
ference of the two types. As appeared later both Wright and Leuckart 
were in error in the interpretation of this organism which is not in any 
sense a sporocyst but a true cercaria although with an unusual type of 
tail. No name has ever been given to this species, which may be desig- 
nated Cercaria wrightti nov. spec. According to the sketch published 
by Leuckart (1886:102) the distome fills three fifths of the stem of 
the anchor, the flukes are nearly straight and together two thirds as 
long as the stem, and the genital organs of the distome form a solid 
rod-like mass located above the acetabulum but partly preacetabular 
and partly postacetabular. The approximate measurements of this 
form taken from the drawing are, total length 0.75, maximum width 
0.133, length of flukes 0.533, breadth of flukes 0.1, length of distome 
0.45, breadth of distome 0.1, diameter of oral sucker 0.041, of ventral 
sucker 0.075 mm. Leuckart speaks of the movement of this species 
as produced by flapping the two wing-like flat fins, evidently depend- 
ing on the notes of Wright, though the latter in his printed notes did 
not refer to the way which the animal moved. 


16 THE JOURNAL OF PARASITOLOGY 


Later M. Braun (1891) published an extended account of a similar 
form which he also found swimming in an aquarium. The material 
came from Kurland. He was able to show that the form was not a 
sporocyst but a true cercaria in which the tail was developed to form 
a receptacle at the anterior end for the body of the young distome. 
He traced the larva back to Limnaea palustris, var. corvus and found 
in the lungs of these snails the numerous sporocysts from which the 
cercariae had come and in which could still be seen all stages in their 
development. He named this form Cercaria mirabilis, and decided 
that it was certainly different from Wright’s species. From his 
description which agrees in general with Wright’s type and with my 
own, some items may be cited to indicate the differences between 
the three. 

Cercaria mirabilis Braun is 6 mm. long with leaflike, movable 
wings, 1.5 mm. long. In resting it lies on the bottom with folded 
wings. In swimming the wings are -moved actively from side to side, 
making the motion resemble that of mosquito larvae. The young dis- 
tome was an opaque yellow body, lying bent in a space lined by a 
smooth membrane. Granules of yellow pigment occur abundantly 
forming a network of lines in the bulb wings, and stalk, and also 
massed in the intestinal crura. The acetabulum is larger than the oral 
sucker. The rudiments of two testes and the ovary lie behind the 
acetabulum. | 

Since the early stages in the development of Cercaria mirabilis, 
taken from sporocysts, are split-tailed cercariae, Braun regarded this 
form as the higher differentiation of that type. He compared it with 
a number of known species but found nothing that furnished a real 
parallel. He was also unable to suggest to what adult distome this 
larval form should be assigned. 

One further point deserves additional emphasis, namely that the 
method of swimming adopted by all three of these forms differs widely 
from that of a typical cercaria. While there are minor differences in 
movement as already noted, yet all accounts agree in stating that the 
forms travel with the wings in advance, trailing behind them the stem 
at the end of which lies the distome in the chamber. The distome is 
so oriented in these cercariae that the anterior end hangs down or back 
and the posterior end is pointed in the direction in which the larva is 
moving. It will be clear on comparing this with the usual cercaria that 
the orientation is precisely reversed; instead of being pushed ahead by 
the tail the distome in this case is pulled along after the tail. This 
is an extremely interesting case of the reversal of functional activity. 


WARD—FREE-LIVING LARVAL TREMATODES 17 


CERCARIA GORGONOCEPHALA 0UV. Spec. 


The other cercaria came from Lake Erie near Put-in-Bay, Ohio. 
It was taken in a tow from a depth of 4 fathoms on July 23, 1901. 
Unfortunately only a single specimen was obtained. Under a dissect- 
ing microscope the object resembled a writhing mass of serpents tied 
together by the tails. The diagrammatic representation (Fig. 3) gives, 
unfortunately, little idea of the actual appearance this object presented 
in life. The bunch contained not less than 50 separate stalks fastened 
firmly together at the base, all in active motion with a spiral twist 
passing in wave-like progression from the base to the outer end of the 
stalk (Fig. 5). The base of each stalk carried a bulbous expansion 
which in some stages of contraction appeared to be sharply cut off 
from the rest of the stalk but at other times graded into the stalk 
without any distinct boundary. This basal enlargement was thicker 
walled than the stalk elsewhere and possessed yellow pigment granules 
that were not found in other parts. The stalk was very mobile, delicate 
in texture and provided with two irregular longitudinal stripes of gran- 
ular brown pigment which terminated just short of the outer tip. This 
stalk which is the region of marked contractile activity, tapered slightly 
(Fig. 4) to the extreme outer tip where it bore the body of a young 
trematode that appeared to be an amphistome. About half of the 
stalks had already lost their attachments; most of these still twisted 
and vibrated nearly as actively as those that carried the young hel- 
minths; but a few were pale in color, appeared empty like dead algal 
filaments, and were motionless. At times all the stalks rested quietly 
for a brief period and then suddenly began to be violently agitated 
together. During this movement it was clear that the stalk alone was 
active whereas the worm was snapped too and fro like the lash of 
a whip. 

As the stalk vibrated with a coiling or twisting motion, the worm 
at the end was turned at every angle and it seemed as if the large 
posterior sucker was mounted on a stalk or at least protruded distinctly 
beyond the general surface of the body. The general form of the 
amphistome was oval with a concave ventral surface and the anterior 
end rolled slightly ventrad (Fig. 6) so that the oral sucker was partly 
concealed. The body of the worm was white, opaque and almost 
entirely immobile. No internal organs could be detected either in the 
attached worms or in those that had been shaken off and lay free in 
the dish. The posterior end of the detached amphistome bore a distinct 
projection which indicated the point at which the stalk had been 
attached. 

A somewhat similar form was discovered by Claus in the Mediter- 
ranean in 1880 and recorded by Leuckart (1886:87). It is referred to 


18 THE JOURNAL OF PARASITOLOGY 


in various other places as a Rattenkonigcercaria. It was carefully 
described by Pintner (1891) under the name of Cercaria clausti given 
it by Monticelli in 1888. Odhner has more recently (1911) shown this 
to be the larval form of Phyllodistomum acceptum Looss. Apart from 
the peculiar habit that both forms are borne on stalks tied in bunches 
there is no close similarity between Cercaria clausit and C. gorgono- 
cephala. The details of structure in the stalk are as unlike as the 
structure of the two worms. 

It is not without some reserve that the species described above is 
assigned to the amphistomes. The form of the body suggests this 
connection but the amphistome cercariae thus far known are very dif- 
ferent in type. Cort (1915) has described two species from this group 
very fully and in common with earlier investigators he finds that two 
eye spots are present in these cercariae. No eye spots were observed in 
Cercaria gorgonocephala. It is possible that the large sucker of this 
cercaria is in reality not posterior but ventral and that the postacetabu- 
lar region is yet undeveloped. No definite opinion on the relationships 
of this form can be given until further material is obtained. 

In one minor structural feature this species manifests a striking 
resemblance to another amphistome cercaria described by Cort. It is 
clear that the stalk is the homolog of the cercaria tail and as can be 
readily seen from an inspection of the sketches published herewith this 
stalk is attached to the young trematode not at the extreme posterior 
tip so as to form a direct posterior extension of the body, but rather 
to the dorsal surface just anterior to the end of the body so that the 
longitudinal axis of the worm when extended lies ventral to that of 
the stalk. This same relation mentioned by Cort in his text is beauti- 
fully illustrated in his sketch of Cercaria diastropha (Pl. 3, Fig. 24). 
The condition exists also in other amphistome cercariae. 

One other point deserves especial notice here. While the stalk 
in this species is in one way an organ of attachment rather than 
a swimming organ, yet it retains its muscular development and struc- 
ture relatively unchanged, and in movement vibrates in a manner which 
suggests to the eye the close similarity to the moving tail of an isolated 
swimming cercaria. One can then not find a basis for calling this in 
a real sense a change of function in the organ. The young distome in 
Cercaria gorgonocephala (Fig. 6) is broader and flatter than any yet 
described in the group of Amphistomata. North American adult 
amphistomes are only very imperfectly known and I am unwilling at 
present to venture a conjecture as to the form to which this cercaria 
belongs. 

The two cercariae discussed in this paper manifest an interesting 
biological similarity. Both differ from the ordinary cercaria in the 
peculiar development of the tail, which has become so highly specialized 


WARD—FREE-LIVING LARVAL TREMATODES 19 


that its original character is not easily discerned. In one case it has 
become a very efficient organ of locomotion, conspicuous both for its 
size and its power; in the other case it has given up its swimming func- 
tion though not its power, and serves to attach the larval worm to 
others in the group. In both cases the end result is the same: The 
larva swimming about in the open water forms a most conspicuous 
object and is readily snapped up by fishes as Braun determined experi- 
mentally with Cercaria mirabilis. The conspicuous character of both 
larvae was also shown in their prompt discovery in the tow although 
they were surrounded by large numbers of other plankton organisms. 
Their activity and conspicuousness must be helpful in enabling them 
to reach a suitable host for further development, and such a host will 
naturally be sought among the fishes of the waters in which the larvae 
occur. 
SUMMARY 


A description is given of the structure and activity of two new 
cercariae of peculiar type captured free in Lake Erie and Lake St. 
Clair. They are designated Cercaria anchoroides nov. spec. and C. 
gorgonocephala nov. spec., and are compared with known European 
species. Altho such an occurrence must be common, these forms are 
the first to be taken in open fresh waters. 


REFERENCES CITED 


Braun, M. 1891. Die sogenannte “freischwimmende Sporocyste.” Centr. 
Bakt. Par., 10: 215-219. 

Cort, W. W. 1915. Some North American Larval Trematodes. Ill. Biol. 
Monogr., 1: 447-532, 8 pl. 

Leuckart, R. 1886. Die Parasiten des Menschen. Leipzig, Bd. 1, Abt. 2, 
Tier... 1. 

Monticelli, F. S. 1888. Saggio di una morfologia dei Trematodi. Napoli, 
130 pp. 

Odhner, Th. 1911. Zum natiirlichen System der digenen Trematoden IV. 
Die Familie Azygiidae n. fam. Zool. Anz., 38: 513-531, 2 figs. 

Pintner, Th. 1891. Ueber Cercaria Clausii Monticelli. Arb. Zool. Inst. 
Wien., 9: 285-294, 1 pl. 

Ward, H. B. 1894. A Preliminary Report on the Worms (mostly parasitic) 
Collected in Lake St. Clair, in the Summer of 1893. Bull. Mich. Fish Com., 
4: 49-54, 1 table. 

Wright, R. R. 1885. A Free-Swimming Sporocyst. Amer. Nat., 19: 310-311. 


: + , ; a ; hy 
i i Ais ‘ uP ' ih Aa ey) 
- ' Je : Ay. i y by nf ’ ‘ i) a Pil m " 
bd ye in Tey)’ q fer ) 
A , f 
Sean, rt lent 


EXPLANATION OF PLATE 


Figs. 1 and 2.—Cercaria anchoroides. 1, young distome just set free, 
2, Cercaria complete, x 105. 


Figs. 3 to 6.—Cercaria garganaecanar Free hand sketches from life. iF r 


details see text. 


= 
>a! 
tas 
. ie 
ey 


ON THE ANATOMY AND RELATIONSHIPS OF SOME 
NORTH AMERICAN TREMATODES * 


Horace W. STUNKARD 


As the result of an extended study of three families of North 
American trematodes, Polystomidae, Aspidogastridae and Paramphi- 
stomidae, certain points of interest in regard to the structure and clas- 
sification have been elucidated. Since the publication of the completed 
work may be delayed, a brief statement of the more important points 
is presented here in advance of the appearance of the extended paper. 

In the latest classification of the monogenetic trematodes, or 
Heterocotylea as they were termed by Monticelli, Odhner (1912) 
divided the group into two suborders, Monopisthocotylea in which a 
“true vagina is present,’ and Polyopisthocotylea in which a true vagina 
is wanting and the so-called “ductus vaginalis” is present. After 
careful study of the female ducts in the Polystomidae, I am able to 
show that the organ which functions as a vagina is homologous in all 
monogenetic trematodes and that there can be no division of the 
group on the basis of differences in this structure. In the complete 
paper the full evidence is submitted to show that the “true vagina” 
cf the Monopisthocotylea is homologous to the originally single, sec- 
ondarily paired and subsequently fused vaginae of the Polyopistho- 
cotylea; altho the two suborders of Odhner are nevertheless valid, 
the essential difference between them is that the genito-intestinal canal 
is lacking in the former and present in the latter group. 

The species that have been included in the genus Polystoma show 
a wider range of structural variation than is usually present in a 
natural genus. There are marked differences in the character of diges- 
tive and reproductive systems and variation exists also in the type of 
adhesive apparatus. In P. integerrimum the ceca are much branched, 
ramifying thru the body and caudal disc. In P. alluaudi the ceca 
occupy the same location but are merely lobed and have no secondary 
branches. In P. bulliense, according to Johnston (1912), “a diver- 
ticulum from the buccal cavity runs backwards, ventral to the pharynx, 
and for a distance equal to its length forming a median unpaired 
buccal pocket.” In all other known species there is a simple bifurcate 
intestine, the ceca terminating just anterior to the caudal disc. In 
two specimens of P. hassalli, however, the ceca are connected 
posteriorly. 


* Contributions from the Zoological Laboratory of the University of Illinois 
under the Direction of Henry B. Ward, No. 72. 


22 THe JOURNAL OF PARASITOLOGY 


The testis is a much branched structure in P. kachugae, in P. inte- 
gerrimum it is lobed, and in the other known species it is oval or 
spherical. In P. integerrimum and P. bulliense the lateral vaginal 
swellings are formed by a large number of papillae which are per- 
forated by fine canals, and in all other known species the vaginae are 
large open funnels and the lateral swellings are reduced or absent. In 
P. integerrimum, P. bulliense and P. alluaudi there is a long uterus 
which forms many loops in the intra-cecal area and contains a large 
number of eggs. In all other known forms, the uterus is situated at 
the level of the ovary on the opposite side of the body and contains a 
single large egg or embryo. 

The caudal disc bears on its ventral face the chief organs of attach- 
ment. These consist of suckers and hooks, the former arranged in 
pairs, three suckers on either side of the median line. In all pre- 
viously reported forms except P. alluaudi, the anterior suckers are 
separated by considerable distance giving the disc the shape described 
by Leidy as cordiform. In the single specimen of P. alluaudi 
described by Beauchamp, both the caudal and cephalic suckers are 
separated while those of each side are contiguous. In P. orbiculare 
n. sp. each sucker of the disc is separated from the two adjacent to 
it by uniform distances, making a perfect circle of bothria. In six 
species studied by the writer these suckers are complicated structures 
set more or less deeply in the parenchyma of the caudal disc. Their 
structure, character of insertion and muscular attachments are 
described in the complete paper. The caudal disc typically bears 
eighteen hooks. The larval hooks are anchor shaped and are situated 
six in a row between the anterior suckers, one inside each sucker at 
the base, and two or four between the posterior suckers. Between the 
posterior suckers there is also a pair of great hooks several times the 
size of the larval hooks, and in species in which a single pair of Jarval 
hooks is present, there is a third pair of hooks similar in shape to the 
great hooks and intermediate in size between the great and larval 
hooks. 

In P. orbiculare n. sp. neither pair of the great hooks are present, 
and in P. opacum n. sp. there is a single pair of great hooks, very 
small and poorly developed. 

The present study of the polystomes has emphasized the morpho- 
logical variation and wide geographic distribution represented by the 
genus. This may mean either that the group is very old and has been 
subjected to conditions producing wide variation, or that it really 
lacks generic entity and consists of various heterocotylean forms which 
have specialized in the direction of an endoparasitic habit and that the 
morphological resemblance is cenogenetic. 

Four new species are added to the genus Polystoma, as follows: 


STUNKARD—ANATOMY OF SOME TREMATODES 23 


POLYSTOMA ORBICULARE Nov. Spec. 


Length 2.7 to 3.75 mm.; width 0.9 to 1.2 mm. Caudal disc circular, 
0.8 to 1.07 mm. in diameter, bothria arranged symmetrically in a circle. 
Only hooks present on disc are larval hooks in bases of suckers. 
Anterior sucker 0.25 to 0.27 mm. in length, 0.37 to 0.42 mm. in width; 
pharynx spherical 0.24 to 0.28 mm. in diameter; esophagus short. 
Testis spherical or oval, 0.36 to 0.5 mm. in length, 0.29 to 0.39 mm. in 
width, near or slightly anterior to middle of body. Genital coronet of 
16 hooks equal in length. Ovary lateral, on either side of body, 
comma-shaped, 0.1 to 0.14 mm. wide by 0.14 to 0.185 mm. long. 
Vitellaria occupy dorsal and lateral areas from pharynx to caudal 
disc except in region dorsal to germ glands where they are reduced or 
absent. 

In the urinary bladder of Pseudemys scripta from Raleigh, N. C., 
and of Chrysemys margmata from Chicago, Illinois, and Creston, 
— Towa. 

POLYSTOMA OPACUM MOV. Spéc. 


Length 3.25 to 4 mm.; width 0.8 to 1 mm. Anterior sucker 0.2 to 
0.22 mm. long, 0.23 mm. wide; pharynx spherical, 0.3 mm. in diameter ; 
esophagus short. Testis spherical or oval, 0.4 to 0.5 mm. in diameter, 
slightly anterior to middle of body. Genital coronet of 32 similar 
hooks. Ovary lateral, comma-shaped or ovoid, 0.16 to 0.2 mm. long, 
0.08 to 0.12 mm. wide. Vitellaria strongly developed, extend from 
pharynx to caudal disc, occupying lateral and dorsal regions of body 
except area over testis, ovary and uterus. 

In esophagus of Trionyx ferox and Malacoclemmys leseuru from 
Newton, Texas. 


POLYSTOMA MEGACOTYLE Nov. Spec. 


Length 2.5 to 2.7 mm.; width 0.71 to 0.78 mm. Caudal disc cordi- 
form; bothria large, overlap. Anterior sucker 0.28 mm. long, 0.35 
to 0.42 mm. wide; pharynx 0.35 to 38 mm. long, 0.38 to 0.44 mm. wide. 
Testis near middle of body, 0.28 to 0.33 mm. long, 0.33 to 38 mm. wide. 
Genital coronet contains 36 hooks in one and 42 in another mounted 
specimen. Ovary broad comma-shaped organ on either side of median 
line, 0.1 mm. long, 0.075 mm. wide. Vitellaria extend in lateral and 
dorsal areas of body from pharynx to caudal disc, reduced or absent 
in small field dorsal to germ glands. 

From oral cavity of Chrysemys marginata, Creston, Iowa. 


POLYSTOMA MICROCOTYLE Nov. spec. 
Length 3 mm.; width 0.78 mm. Caudal disc cordiform; bothria 
small, separated. Anterior sucker 0.2 mm. long, 0.42 mm. wide; 
pharynx 0.37 mm. long, 0.4 mm. wide. Testis slightly anterior to 


24 THE JOURNAL OF PARASITOLOGY 


middle of body, 0.36 mm. long, 0.42 mm. wide. Genital coronet of 

32 hooks, equa] in length. Ovary lateral, 0.075 mm. long, 0.1 mm. 

wide. Vitellaria well developed, same extent as in P. megacotyle. 
From oral cavity of Chrysemys marginata, Creston, Iowa. 


In the family Aspidogastridae the three North American species 
have been restudied. A detailed comparison of specimens of Aspido- 
gaster conchicola with the descriptions of Voeltzkow (1888), Stafford 
(1896), and other writers confirms former observations and sub- 
stantiates the statements of Leidy (1851) and subsequent authors that 
A. conchicola occurs in this country. The examination of specimens 
of Cotylaspis insignis and Cotylaspis cokeri corrects and supplements 
former descriptions. Nickerson’s (1902) classification of the family 
is revised and brought to date. 


Representatives of three species of paramphistomes have furnished 
the basis for studies on that family. Two species are from North 
American turtles and the third is from a duck, Anas platyrhynchos. 
An examination of the literature showed that these forms could not 
be included in any previously described genera. 

A new genus Alassostoma is created to contain the two species 
from turtles. The genus is characterized by the presence of large oral 
evaginations which open separately from the oral sucker, an esopha- 
geal bulb composed of concentric muscle lamellae, germ glands situated 
near the middle of the body in the median line, both testes anterior to 
the ovary, vitellaria consisting of small scattered follicles in the lateral 
and posteriorly in the median area of the body; Laurer’s canal opens 
in the mid-dorsal line, anterior to the opening of the excretory vesicle; 
cirrus sac and uterus open to the exterior thru a common hermaphro- 
ditic duct. Alassostoma magnum n. sp. is taken as type of the genus 
in which is included also Alassostoma parvum n. sp. 

The genus Alassostoma has the type of lymph and excretory 
systems present in the genus Schizamphistoma and designated by 
Looss (1912) as characters of the subfamily to which that genus 
belongs. Looss predicted that with the discovery of other genera it 
would be necessary to create a new subfamily to contain them, and at 
that time stated the subfamily characters. With the discovery of a 
second genus so similar to Schizamphistoma, the formal erection of the 
new subfamily is necessary. Schizamphistoma Looss was designated 
as the type genus and the name of the subfamily becomes Schizamphis- 
tominae. The distinguishing characters of the subfamily are stated 
by Looss to be two long excretory vesicles which extend singly to 
the anterior end of the body and a lymph system composed of three 
canals on either side of the body which extend longitudinally and 


——— << " 


STUNKARD—ANATOMY OF SOME TREMATODES 25 


break up into many sinuses in the region of the suckers. The sub- 
family contains the genera Schizamphistoma, including also S. spinu- 
losum which as indicated by Looss should probably be made the type 
of a new genus, and the genus Alassostoma. 


Alassostoma magnum agrees with Schizamphistoma scleroporum 
in general appearance and size, in type of excretory and lymph sys- 
tems, in character of vitellaria, and in general type of reproductive and 
alimentary organs; but A. magnum has large oral evaginations, which 
pockets are reduced and do not extend outside the sucker in S. sclero- 
porum,; further A. magnum lacks the preoral sphincter which is present 
in S. scleroporum. In A. magnum the uterus and cirrus sac open 
to the surface thru a common hermaphroditic duct; in S. scleroporum 
they open separately. In A. magnum the testes are further porteriad 
and the ovary is situated one fourth to one third of the body length 
from the posterior end instead of at the level of the anterior margin 
of the acetabulum as is the case in S. scleroporum. In the latter species 
the testes and ovary are widely separated whereas in A. magnum they 
are relatively close together. A. magnum agrees with S. spinulosum 
in the presence of oral evaginations and lack of preoral sphincter, but 
differs from it in the manner of coiling of the excretory vesicles, in 
the presence of common hermaphroditic duct, in the character of the 
vitellaria, as well as in relative positions of the testes and ovary. 
These morphological data show differences too fundamental to permit 
the inclusion of A. magnum in the same genus with either S. sclero- 
porum or S. spinulosum. 


Alassostoma parvum agrees with A. magnum in general morpho- 
logical features, presence of oral evaginations, lack of preoral sphinc- 
ter, type of lymph and excretory systems, in character of genital organs 
and ducts, also in relative position of testes and ovary. A. parvum 
therefore agrees with and differs from S. scleroporwm and S. spinu- 
losum in the same manner as A. magnum. That the two American 
forms are not different developmental stages of the same species is 
shown by the great difference in size of the worms and relative differ- 
ences in size of suckers and genital organs. One mounted specimen 
of A. magnum 10 mm. long is not sexually mature while none of the 
individuals of A. parvum are more than 3 mm. in length. A. magnum 
is large with small suckers, whereas A. parvum is small with relatively 
large suckers; and this feature suggested the name Alassostoma. 

Alassostoma magnum was collected from the large intestine of 
Pseudemys troosti and P. elegans from Havana, Illinois, and from 
P. elegans from Chicago, Ill. The specimens of A. parvum were found 
in the cloaca of Chelydra serpentina at Urbana, III. 


26 THE JOURNAL OF PARASITOLOGY 


The paramphistomes from Anas platyrhynchos were collected in 
Rock County, Nebraska. Unfortunately the fixation of the parasites 
is such that the excretory and lymph systems can not be traced, altho 
remnants of both appear in sections. This species closely resembles 
Amphistoma lunatum Diesing. Both are parasites of American ducks, 
and are the only paramphistomes at present known from avian hosts. 
They are nearly equal in size, are similar in shape, have a subterminal 
oral sucker, reproductive and digestive systems that compare closely, 
and acetabula of the same form, consisting of an anterior portion and 
a posterior overhanging lip which terminates on either side in a small 
cone-like projection. The species at hand differs from A. lunatum 
in its smaller oral evaginations, shorter esophagus, and in having oval, 
lobed testes and ovary instead of spherical germ glands. The acetabu- 
lum is nearer the ovary and the vitellaria are entirely extracecal while 
in A. lunatum they extend between the ceca. 

Amphistoma lunatum has been placed as an appendix to every 
classification of the paramphistomes that has ever been attempted. 
With the discovery of a form so similar that the two. must belong 
together, a new genus is proposed to contain the two species. The 
peculiar divided condition of the acetabulum suggested the name 
Zygocotyle for the genus. The species at hand, for which I propose 
the name Zygocotyle ceratosa, is designated as type and in the genus 
is included also the species Z. lunatum (Diesing). As diagnostic 
characters of the genus may be mentioned the subterminal oral sucker, 
posterior sucker divided or provided with a caudal overhanging lip, 
absence of cirrus sac, and separate openings for male and female 
ducts. Others will undoubtedly appear when the character of the 
excretory and lymph vessels are known. The genus Zygocotyle differs 
from all other known genera of the Paramphistomidae in the ventral 
position of the oral sucker and the peculiar character of the acetabu- 
lum. None of the existing subfamilies will include it fairly, but since 
the present classification of the Paramphistomidae is somewhat uncer- 
tain and the structure of the lymph and excretory systems of this 
genus is as yet unknown, no further attempt at classification of the 
group is made at this time. 7 

Types of all the new species described in this paper have been 
deposited in the Helminthological Collection of the University of 
Illinois. 

SUMMARY 


Extended study of North American representatives of the three 
trematode families, Polystomidae, Aspidogastridae and Paramphi- 
stomidae has made possible the first comprehensive treatment in this 


STUNKARD—ANATOMY OF SOME TREMATODES 27 


country of their structure and classification. Four new species are 
added to the genus Polystoma and three new species of two new 
genera are added to the Paramphistomidae. 


LITERATURE CITED 


Johnston, S. J. 1912. On Some Trematode Parasites of Australian Frogs. 
Proc. Linn. Soc. N. S. Wales, 37: 285-362, pl. 14-43. 

Leidy, J. 1851. Helminthological Contributions II. Proc. Acad. Nat. Sci. 
Phila., 5: 224-227. 

Looss, A. 1912. Ueber den Bau einiger auscheinend seltener Trematoden- 
Arten. Zool. Jahrb., Suppl., 15: 323-366, 3: pl. 

Nickerson, W. S. 1902. Cotylogaster occidentalis n. sp. and a Revision of 
the Family Aspidobothridae. Zool. Jahrb., Syst., 15: 597-624, 2 pl. 

Odhner, T. 1912. Bemerkungen zum nattirlichen System der Monogenen 
Trematoden. Zool. Anz., 39: 337-351. 

1913. Noch einmal die Homologien der weiblichen Genitalwege der mono- 
genen Trematoden. Zool. Anz., 41: 558-559. 

Stafford, J. 1896. Anatomical Structure of Aspidogaster conchicola. Zool. 
Jahrb., Anat., 9: 477-542, 4 pl. 

Voeltzkow, A. 1888. Aspidogaster conchicola. Arb. zool.-zoot. Inst. Wiirzb., 
8: 249-292, 5 pl. 


DAUERCYSTFORMATION OF TRICHOMONAS 
INTESTINALIS * 


KENNETH M. Lyncu, CHARLESTON, S. C. 


Ucke (1908), Bohne and Prowazek (1908), and Bensen (1910), 
have described encystment of Trichomonas intestinalis. Bensen 
(1910) has also described an encystment for Trichomonas vaginalis 
differing from that of Trichomonas intestinalis, and Dobell (1908) 
reports a dauercyst of Trichomonas batrachorum. Alexeieff (1911) 
disputes the nature of the so-described cyst of Trichomonas intestt- 
nalis, asserting that it is in reality an ascomyces, a vegetable organism 
akin to the yeasts, and proposes for it the name Blastocystis entero- 
cola. Wenyon (1905) and others uphold Alexeieff’s contention, 
Wenyon calling the organism Blastocystis honunis. 

The question of encystment of Trichomonas intestinalis 1s an inter- 
esting one to me because of the questioned nature of the cyst which 
has been previously described and because of the prevalence of the 
parasite in this community. For several years I have been observing 
Trichomonas as a parasite of several locations in the human body and 
in certain lower animals, and have frequently encountered a form 
which has proven to be a distinct cyst and not to be confused with 
that previously described as an encysted Tr. intestinalis by Ucke, Bohne 
and Prowazek, and Bensen, and as Blastocystis enterocola by Alexeieff. 

At the present I have under observation a man who furnishes this 
cyst in large numbers and with distinct characteristics. This man is 
a negro who is in the hospital with chronic endocarditis and who gives 
no history of dysentery. 

In the stool no other protozoon has been found, no cell corre- 
sponding to the previously described Tr. intestinalis cyst and none of 
the Blastocystis of Alexeieff. There are many active Trichomonas 
conforming to the typical organism, averaging about 8 by 12 micra in 
size, of elongated pear shape, with constant undulating membrane, 
three flagella, the stiff spine projecting posteriorly, vacuolated cyto- 
plasm with numerous ingested bacteria, and with nucleus indistinct but 
showing well in stained preparations. On exposure the active form 
soon becomes inactive, stationary, and does not develop the irregular 
ameboid, non-flagellated, undulating form. It is one of the most 
fixed types I have observed. 


* From the Department of Pathology and Research Medicine of. the Medical 
College of the State of South Carolina. 


ea rcC lr 


LYNCH—CYSTS OF TRICHOMONAS 29 


The encysted form is almost as numerous as the active and com- 
monly exhibits a tendency to occur in pairs. It is about three-fourths 
the size of the active, of a typical pear shape, and has a transparent 
shell of uniform thickness. The enclosed parasite has a regular ovoid 
contour and a finely granular grayish appearance. On one side nearer 
to the small end the nucleus is visible as a refractive granule, and on 
the other the undulating membrane is seen as a refractive wavy line 
extending from end to end. 

More definite observations of this cyst and its different structures 
may be made from stained specimens. The technic of staining which 
I have used is as follows: A thin spread of the feces while still wet is 
fixed in warm saline alcoholic corrosive sublimate for ten minutes, 
placed in absolute alcohol ten minutes, iodin solution ten minutes, 
alcohol ten minutes, washed in distilled water, mordanted in 4% 
aqueous ferric ammonia alum overnight, stained in 1% alcoholic 
hematoxylin one day, decolorized in 2% aqueous ferric ammonia alum, 
counterstained in alcoholic eosin and cieared in carbol-xylol. 

In such a preparation the encysted form is more deeply stained 
than the active. It is about 6 by 8 micra in size and of perfectly 
symmetrical pear shape. The anterior end projects on a shoulder 
slightly beyond the general line; the wall is uniformly distinct; and 
the space between the cyst wall and the body of the parasite is usually 
distinct and clear, it being usually broader at the anterior end. 

The body of the enclosed parasite is of symmetrical, ovoid shape, 
slightly pointed anteriorly, of dull brick-red color, finely granular and 
contains no vacuoles or food particles. A fine dark line beginning as 
a granule in the anterior end runs directly backward to the posterior 
end. This I take to be the stiffening rib of the undulating membrane 
because of its close association in origin and termination with that 
organ. The undulating membrane is distinct as a darker line begin- 
ning in close connection with this rib and extending backward along 
one side of the body in a wavy course to the posterior end, where it 
curves around the extremity of the parasite and comes to end near 
this end of the rib. The nucleus is comparatively large, of ovoid form, 
but lies farther back than in the active parasite and on the side opposite 
to the undulating membrane. Its usual position is in the posterior 
part of the anterior half, and between the midline and the body wall. 
It has a distinct dark-stained rim and a large chromatin mass. This 
chromatin usually occurs as an irregular black body almost filling the 
nucleus, but is in some broken into smaller granules and in others 
distributed around the inner edge of the nuclear rim. In addition to 
the undulating membrane the cyst usually shows two or three more 
delicate lines arising in close association with that organ and passing 
backward over the body for about two thirds of its length. These are 


30 THE JOURNAL OF PARASITOLOGY 


probably flagella. They stain poorly and are not constantly seen, 
especially in the more faintly stained specimens. The characteristics 
of the fully formed cyst may be seen in Figure A3 which is an off- 
hand drawing. 

In addition to this fully developed cyst which predominates, there 
are young cysts and forms of apparent pre-encystment. There is a 
form which is smaller than the active parasite, shorter and more blunt, 
with cytoplasm somewhat reticulated, but showing no vacuoles and 
no bacteria or other ingested materials, and with nucleus more distinct 
and with larger amounts of chromatin. This I take to be a pre- 
encysted stage. Figure Al appears to be a young cyst before the wall 
has reached full formation. Its shape is typical; the body wall is 
thick; and the internal organs appear as in the fully encysted. A 
further stage appears to be the form in which there is a space between 
the shell and parasite only at the anterior end, these parts being in 
immediate contact around the remainder of the body, see A2. 


Fig. Al, 2, and 3—Drawings of different stages of dauercysts of Tricho- 
monas intestinalis as seen in specimens stained in hematoxylin. 


Further development suggestive of multiplication I have not seen 
in these cysts; and the preservation of the undulating membrane and 
flagella, together with the single nucleus, indicate that the process is 
not for reproduction but simply for resistance. 


DISCUSSION 


According to these observations the formation of a resistance cyst 
plays a part in the life of Trichomonas intestinalis as occurs with other 
intestinal protozoa, and I believe is the stage in which the parasite may 
be transmitted. That infection by the contracted but non-encysted 
parasite occurs I am not prepared to dispute, and it seems probable to 
me that the contracted form may be a pre-encystment stage. Reason- 
ing by analogy leads one to believe that infection through the stemach 
by means of the non-encysted is not probable, and certain observations 
which I have made of the purely active Trichomonas from the vagina 
also rule against such a manner of infection. 


LYNCH—CYSTS OF TRICHOMONAS 31 


I have previously reported (1915) a case of vaginal and mouth 
infection by Trichomonas which did not infect the intestine, this being 
determined by repeated examination after purging. I have since had 
a similar case of trichomoniasis of the vagina and mouth; and again 
by repeated antemortem and also postmortem examinations failed to 
find the intestine infected. In these cases the parasites seemed iden- 
tical in the two situations, and there was never any but the pure fixed 
type of active form. There were enormous numbers in the mouth for 


Fig. B1, Dauercyst of Tr. batrachorum by Dobell; 2, Encysted Tr. vaginalis 
by Bensen; 3, Cyst of Tr. intestinalis by Wenyon; 4, Encysted Tr. intes- 
tinalis by Ucke; 5, Encysted Tr. intestinalis by Bohne and Prowazek; 
6, Encysted Tr. intestinalis by Bensen; 7 and 8, Blastocystis enterocola by 
Alexeieff ; 9, Blastocystis hominis by Wenyon. 


considerable periods of time; and if the active Trichomonas is capable 
of transmission through the stomach to the intestine in man, the 
swallowing of these organisms should have produced an intestinal 
infection in these women. 

It is not my purpose at this’ time to enter into a discussion as to 
the nature of the previously described encysted Tr. intestinalis which 
has been called Blastocystis by Alexeieff; but from extensive observa- 


32 THE JOURNAL OF PAKASITOLOGY 


tion of both organisms, both associated and occurring separately, I 
am in. accord with the view that it is not a Trichomonas cyst. 

In order that the lack of resemblance of this cell to the cyst here 
described may be seen I have included copies of figures by Ucke, Bohne 
and Prowazek, Bensen, Alexeieff, and Wenyon. Figures B5 and 6 are 
from a hematoxylin-stained specimen. Hence the main difference in 
appearance to Figures B4, 7, 8, and 9, which are representations of the 
unstained cell, since the internal part of the cell, which is often trans- 
parent in the fresh specimen, stains rather deeply and the nuclei are 
more distinct in the stained. Figures B4, 5, and 6 are illustrations of 
the so-called Trichomonas intestinalis cysts of Ucke, Bohne and Pro- 
wazek, and Bensen. Figures B7, 8, and 9 are illustrations of 
Blastocystis by Alexeieff and Wenyon. There is seen to be no point 
of similarity between the Trichomonas cyst of Ucke, Bohne and Pro- 
wazek or Bensen and that here described and pictured in Figure A; 
whereas, barring differences in preparation and in stage of the organ- 
ism, the Blastocystis of Alexeieff and Wenyon and the Trichomonas 
cyst of Ucke, Bohne and Prowazek and of Bensen are apparently one 
and the same organism. 

There is however some resemblance between this cyst and Bensen’s 
encysted Trichomonas vaginalis (Figure B2) and Dobell’s cyst of 
Trichomonas batrachorum (Figure Bl), which I have taken the 
liberty of copying for comparison. In the last two the flagella are pre- 
served until the cyst is formed, but lost afterwards; while the typical 
shape of the Trichomonas intestinalis cyst is not observed, and the 
undulating membrane and axostyle are not seen. The nucleus is also 
different, in that in 77. intestinalis it is more rounded and placed more 
posteriorly, while in both Tr. batrachorum and Tr. vaginalis it is 
forward, larger and spindle shaped. Then in the case of Tr. vaginalis 
Bensen has described and illustrated further development of the cyst 
for multiplication, while further development of the cyst here 
described has not been seen. 


CONCLUSION 


Accordingly, therefore, the formation of a resistance cyst does 
play a part in the life of Trichomonas intestinalis, and this cyst bears 
no relationship to the cell which has been previously described as an 
encysted Tyr. intestinalis by Ucke and others, and as Blastocystis 
enterocola by Alexeieff, and differs essentially from Dobell’s dauer- 
cyst of Trichomonas batrachorum and Bensen’s encysted Tr. vaginalis. 


SUMMARY 


Encystment of Trichomonas imtestinalis has been investigated for 
many years. That which has been previously described has not stood 


LYNCH—CYSTS OF TRICHOMONAS 33 


the test of investigations The morphology of the cyst here described 
identifies it with the parasite from which it arises. It is not to be 
confused with any cell occurring in the intestine and feces. Whether 
this cyst releases more than one organism or whether Trichomonas 
intestinalis has a multiplication cyst remains an unanswered question. 


REFERENCES CITED 


Alexeieff, A; 1911, . Sur la nature des Formations Dites “Kystes de 
Trichomonas intestinalis.” C. R. Soc. Biol., 71: 296-298, 

Bensen, W. 1910. Trichomonas intestinalis und vaginalis des Menschen. 
Arch. Protist., 18: 115-127. : 

Bohne, A., und Prowazek, S. V. 1908. Zur Frage der Flagellatendysenterie. 
Arch. Protist., 12: 1-8. 

Dobell, C. C. 1908. Trichomonas batrachorum Perty. Proc. Cambridge Philos. 
Soc., 14: 428-433. 

Lynch, K. M. 1915. Trichomoniasis of Vagina and Mouth. Amer. Jour, 
Trop. Diseases and Prev. Med., 2: 627-634. 

Ucke, A. 1908. Trichomonaden und Megastomum ‘im Menschendarm. 
Centralb. Bakt. Par., I. Abt., 45 : 231-233. 

Wenyon, C. M. 1905. The Common Intestinal Protozoa of Man: Their 
Diagnosis and Pathogenicity. The Lancet, London, 189: 1173-1183, 


NOTES “ON PWO' CESTODES FROM “THE: 'SPOTTES 
STING-RAY 


EpwIn LINTON 


A single specimen of a species of cestode found in the spiral valve 
of a cow-nosed ray (Rhinoptera bonasus) at Woods Hole, July 29, 
1887, was made the type of a new genus and species (Tylocephalum 
pingue). No other examples of this genus have been found at Woods 
Hole, but on June 30, 1908, at the Tortugas laboratory, I obtained two 
specimens of a cestode from the spotted sting-ray (Aetobatis narinart) 
which are to be referred to the genus Tylocephalum. The specimen 
from the cow-nosed ray was a less mature strobile than those from the 
spotted ray; a comparison of the genitalia, therefore, cannot be made. 
There appears to be enough difference, however, in other particulars to 
justify referring the Tortugas specimens to a new species. While both 
hosts belong to the family of eagle rays, there is enough difference 
between them in the way of geographical range and generic features 
to make it unlikely that the same species of cestodes should be found 
in each. 

TYLOCEPHALUM MARSUPIUM Nov. spec. 


‘Scolex: The relatively large, muscular portion (myzorhynchus) is 
subglobular, its length in a living specimen 0.16 and breadth 0.21 mm.; 
bothria united into a subglobular disc with four auxiliary acetabula, 
length of disc 0.30, breadth 0.69 mm. The constriction noted in the 
Woods Hole specimen not present. As in the case of the specimen from 
the cow-nosed ray, the scoleces were rather firmly fastened to the 
mucous membrane of the spiral valve. One of the worms was fixed 
without detaching it, and was sectioned together with a small piece 
of the intestinal wall. The sections show that the myzorhynchus 
alone had entered the mucous membrane. 

Strobile: The segments begin nearer the scolex than they do in 
T. pigue. Just behind the scolex, where the breadth was 0.16, the 
strobile was crossed by crowded lines. One-half millimeter back of 
the scolex the well-defined segments were 0.014 mm. in length and 
0.18 mm. in breadth. Three millimeters back the length of the seg- 
ments is about 0.05 and the breadth 0.24; ten millimeters back the 
length is 0.12, the breadth 0.28; twenty millimeters back the length is 
0.28, the breadth 0.24; thirty millimeters back the length is 0.46, the 
breadth 0.28; forty millimeters back of the scolex the length is 0.56, 
the breadth 0.38 mm. The last segments are somewhat variable in 
their dimensions, but are about one millimeter in length and 0.5 mm. 


LINTON—CESTODES FROM STING-RAY a5 


in greatest breadth. They are vase-shaped, constricted at the anterior 
end, swelling out to the maximum breadth behind the middle, slightly 
constricted near the posterior end with a moderately projecting poste- 
rior margin. One proglottis had the following dimensions: Length 
0.84; breadth, anterior 0.21, maximum 0.56, posterior 0.39 mm. The 
strobile is especially distinguished by the strongly developed longi- 
tudinal muscles. The longitudinal muscles are disposed in radial 
bundles near the scolex (Fig. 1), but farther back lie in a well-defined 
zone (Fig. 2). In segments in which the genitalia have become dif- 
ferentiated this zone of muscle bundles coincides in position with the 
vitellaria (Fig. 4). 


B 


Text Fig. A——Tylocephalum marsupium. View of scolex in life, somewhat 
flattened and seen from behind. Breadth of scolex 0.7 mm. 


Text Fig. B—Onchobothrium tortum. Side view of scolex; balsam. Diame- 
ter at base of hooks 0.64 mm. 


Genitalia: The general plan of arrangement of the genitalia is 
shown in Figure 7. The vitellaria are peripheral and consist of 
rather finely granular masses lying between and also centrally to the 
muscle bundles. The testes are in the median region. In the younger 
proglottids they occupy most of the interior, but as the proglottids 
mature they give way to the seminal receptacle and ovary. The 
cirrus-pouch is relatively small and oval, opening near the margin not 
far from the middle of the length. The vagina opens into the genital 
cloaca, passes along one side of the cirrus pouch, becomes more or 
less convoluted and expands into a capacious seminal receptacle: This 
was filled with spermatozoa in all the later proglottids. The ovary 
is lobed and is situated at the posterior end of the proglottis. 

The uterus was still rudimentary even in the mature proglottids. 
In a section a small cluster of minute bodies was seen. They lay in 
‘the lumen of the uterus, were yellowish brown, and about 0.010 by 
0.007 mm. in the two principal diameters. 


36 BE JOURNAL OF PARASITOLOGY 


ONCHOBOTHRIUM TORTUM Nov. Spec. 


Ten specimens of this form were obtained from a spotted sting- 
ray (Aetobatis narinari), June 30, 1908. The scolices were imbedded 
in the intestinal wall and had caused some ulceration. One of the 
worms, straightened out on a glass plate in sea water, measured 220 
mm. in length. Anterior end sub-cylindrical, with a tendency to coil 
spirally; color dark ashy-gray. Scolex long-clavate, armed with four 
pairs of short, sharp, two-pronged hooks. Each pair of hooks situated 
at the anterior end of one of the four bothria. The latter are oblong, 
trough-shaped, with two coste near the posterior end. Behind the 
scolex the body is at first sub-cylindrical and crossed by fine, closely 
crowded lines for a considerable distance. The segments outlined by 
these transverse lines remain closely crowded, while the adult pro- 
glottids begin rather abruptly. The average length of the first 12 
adult proglottids was 0.8 mm., the breadth being about the same or 
slightly greater. The diameter of the sub-cylindrical portion of the 
strobile was about 1.5 mm. The scolex and anterior portion of the 
strobile are much thicker than the adult proglottids. Diameter of 
scolex, in alcohol, anterior 0.85, middle 0.77; diameter of neck, a short 
distance back of the scolex, 1.4 mm. Dimensions of one of the pos- 
terior proglottids: life, length 1.47; breadth, anterior 0.5; middle 0.8, 
posterior 0.6 mm. Dimensions of scolex mounted in balsam: length 
0.97; breadth, at base of hooks, 0.97, behind hooks, 0.81; breadth of 
neck, a short distance behind the scolex, 1.27 mm. In the mounted 
specimen the neck is seen to be traversed by strong longitudinal muscle 
bundles which are closely crowded together, each bundle about 0.06 
mm. in diameter. About 16 bundles were counted near the head; 
farther back they are divided into a larger number of smaller bundles. 
Two spiral vessels show distinctly in the mounted specimen. The 
strobile narrows as the proglottids become distinct. In the specimen 
which measured 220 mm. there were distinct and well-formed segments 
on the last 150 mm. The maturing segments were at first much 
broader than long, then squarish, then longer than broad, the last ones 
three times as long as broad. The posterior margins of the pro- 
glottids project slightly and have crenulate borders. One of the 
posterior proglottids of a mounted strobile has the following dimen- 
sions: length 1.86; breadth, anterior 0.36, constriction near anterior 
end 0.25, middle 0.40, posterior margin 0.54 mm. The genital aper- 
tures are marginal at about the middle of the length. They are irregu- 
larly alternate. No ova were seen. 

The general plan of arrangement of the genitalia is shown in 
Figure 8. The cirrus is armed with slender, bristle-like spines; a few 
folds of the vas deferens are included in the oval cirrus-pouch at its 


LINTON—CESTODES FROM STING-RAY 37 


medial end. The voluminuous folds of the vas deferens form the 
seminal vesicle and occupy the median third of the anterior half of 
the proglottis. The testes are situated in the anterior half of the pro- 
glottis, and occupy the median region on each side of the vas deferens. 
On the marginal sides of the testes are the vitelline glands which extend 
along each marginal border of the entire length of the proglottis, 
being interrupted only at the point where the cirrus pouch and the 
accompanying vagina approach the genital aperture. The uterus was 
represented by a tubular structure lying along the median line near one 
of the lateral faces of the proglottis, and extending from nearly one 
end of the proglottis to the other. The ovary is a lobed organ and fills 
all the space between the marginal vitellaria behind the cirrus pouch. 
The vagina opens at the genital pore immediately in front of the cirrus 
and lies alongside the anterior border of the cirrus pouch. At this 
point it is thick-walled and glandular. It becomes tubular at about 
the level of the median end of the pouch and passes along the median 
line beneath the uterus to about the middle of the ovary. The relative 
positions of vagina and uterus are shown in Figure 9, which is sketched 
from a transverse section of a maturing segment at a level which 
passes very near the genital aperture, shows a portion of the vagina 
near the margin, cuts into some folds of the vas deferens, and passes 
thru the vagina again near the middle of the segment, where it lies 
on the medial side of the uterus. The section also catches a few of 
the anterior lobes of the ovary. In this section the characteristic 
longitudinal muscles are seen as an inner circle of larger and an outer 
circle of smaller bundles. The lateral vitellaria and the median testes 
flanking the folds of the seminal vesicle are also shown . 


SUMMARY 


Two new species of cestodes, of the genera Tylocephalum and 
Onchobothrium, respectively, are described in this paper. One of 
them, 7. marsupium, is the first cestode of this genus to be recorded 
since the genus was established in 1887. Thus far representatives of 
this genus have been found only in the eagle rays. 

Altho the two genera belong to quite different families, they pos- 
sess an interesting feature in common in the strongly fasciculated 
longitudinal muscle layers. Both species were fastened to the mucous 
membrane of the spiral valve which, at the point of attachment of the 
onchobothria, was somewhat ulcerated. 


EXPLANATION OF PLATE 


. 


' Fig. 1—Tylocephalum marsupium. Transverse section of neck. Diameter 
0.22 mm. 

Fig. 2—Tylocephalum marsupium. Transverse section of early proglottis, 
showing rudiment of genitalia and peripherally arranged longitudinal muscle 
bundles. Greater diameter 0.65 mm. 

Fig. 3—Onchobothrium tortum. Transverse section of neck, showing longi- 
tudinal muscle bundles and vessels of the vascular system. Longer diameter of 
section 1.12 mm. 

Fig. 4—Tylocephalum marsupium. Transverse section of mature proglottis 
in front of cirrus bulb; longer diameter 0.45 mm. 

Fig. 5—Onchobothrium tortum. Longitudinal view of neck showing muscle 
bundles. Breadth 1.17 mm. 

Fig. 6—Onchobothrium tortum. View of retracted cirrus, and vagina; from 
longitudinal section. 

Fig. 7—Tylocephalum marsupium. Posterior proglottis; outline from life; 
genitalia partly diagrammatic. Length 0.8 mm. 

Fig. 8.—Onchobothrium tortum. Posterior proglottis; balsam. Length 
1.6 mm. 

Fig. 9—Onchobothrium tortum. Transverse section of a somewhat younger 
proglottis than that shown in Figure 8. Longer diameter of section 1.12 mm. 

Fig. 10—Onchobothrium tortum. Details of musculature. 


ABBREVIATIONS USED 


c, retracted cirrus t, testes 

cm, circular muscle layer u, uterus 

gp, genital pore v, vagina 

Im, longitudinal muscle bundles vd, vas deferens 
0, ovary vg, vitellaria 


sr, seminal receptacle w, longitudinal vessel 


“ 


PLATE 


a 
A ; 
1b estate ore, 
ees l 


or 


Siatita., 
US 


Fas 
oe 


4 br RAIN F 
RR ow HE ae 


TN IAUA ZT 
Z\TNIES 


—= 
AY 


PA BINGE 


®& CASE OF THE OCCURRENCE OF ASCARIS TRI- 
QUETRA SCHRANK IN DOGS * 


A. C. WALTon 


While working on the spermatogenesis of certain Ascaridae last 
year, I found that the chromosomes of the ascarids from dogs did not 
agree with those of the ascarids from the dog as given by Kultschitzky 
(1888) and by Marcus (1906) either in number, behavior, or the 
presence of an idiochromosome group. The work of Glaue (1908, 
1909, 1910) has shown conclusively that the ascarids of the dog and 
of the cat are anatomically distinct species, which should be designated 
respectively as Ascaris canis Werner, and Ascaris felis Goeze, and 
not merely varieties of Ascaris mystax Zeder. The work of Edwards 
(1911) on A. felis and that of Marcus (1906) on A. canis have given 
us conclusive evidence that these two forms are entirely dissimilar as 
to the number and the behavior of the chromosomes. From these 
taxonomic and cytological proofs, the long mooted question of the 
identity of the two varieties seemed definitely settled ; but the apparent 
contradiction in the gametogenesis of the dog ascarids shown by my 
discovery seemed to me sufficient to warrant the reopening of the 
question. If the number and behavior of the chromosomes in Ascaris 
canis were similar to the number and behavior of those in Ascaris felis, 
the two forms might be in fact only sub-species ; varying taxonomically 
owing to their different environments. 

The results of my study are contained in a paper now in press, 
the taxonomic work of which showed that the species with which | 
was working were the ones recognized by helminthologists as the 
usual inhabitants of the intestine of the dog and the cat, respectively. 
My work on the gametogenesis of A. felis agreed with that of Edwards 
(1911) in showing nine chromosomes as the haploid number, one of 
which is a member of an X-Y idiochromosome pair. 

Marcus (1906) has shown that what he called A. canis has ten 
paired and two unpaired tetrad chromosomes as the diploid number. 
From his description it seems probable that these two unpaired 
chromosomes act as members of an X-Y idiochromosome group, but he 
did not so call them. My work on the commonest parasite of the dog 
has shown that in the male there are thirty tetrad chromosomes as the 
diploid number, of which twenty-four are united in pairs, and the 


* Contributions from the Zoological Laboratory of the Museum of Compara- 
tive Zoology at Harvard College, No. 283. 


40 THE JOURNAL OF PARASITOLOGY 


other six form a heterochromosome group of the X type. The female 
has thirty-six tetrads (eighteen di-tetrads) as the diploid number. 

Private correspondence between Dr. S. I. Kornhauser, of North- 
western University, and Dr. Marcus has shown that the majority of 
the material upon which the latter based his work was not obtained 
from dogs, but came mostly from other members of the dog family 
and also from bears. 


it 2 

Fig. 1—Dorsal aspect of the anterior end of Ascaris triquetra Schrank 
(SC25): 

Fig. 2—Same view of Ascaris canis Werner (X25). 

Fig. 3.—Cross-section. Posterior aspect of the right wing of A. triquetra 
(X 160). .. 

Fig. 4—Same for A. canis (X 160). 


Fig. 5.—Lateral view of the right side of posterior end of male A. triquetra 
(X 25). 
Fig. 6—Same view of male A. canis (X25). 


All drawings were made with the aid of a camera lucida. 


During the past two years I have been able to examine worms 
taken from twenty-five dogs, and of the total of two hundred worms 
thus obtained, all but two have answered taxonomically and cytologic- 
ally to the type described above as the commonest Ascaris in dogs, 
i. e., Ascaris canis Werner. These two exceptional specimens, a male 


WALTON—ASCARIS TRIQUETRA 41 


and a female of the same species, differed considerably in taxonomy 
from the ordinary type of Ascaris canis Werner. The following table 
compares the main features of the two forms: 


A. canis Werner A. triquetra Schrank 

Feneth of male...:....... WAY tare nar: sence 60 mm. 
Length of female......... TAQ ime see ete es 100 mm. 
Shape of oral wing........ Manceolate estas ae Broadly lanceolate 
himemsess of oral wine... 0.17 mame. 9.0.4.5... = 0.18 mm. 
Breadth of oral wing...... O:GS naa ete eho ns 0.18 mm. 
Length of oral wing....... BF Mn ee ee ae tte ce + 1.9 mm. 
Chitin rod of wing........ Long and: broads... =: Shorter and narrower 
Post-anal: papillae: . <2. 40. < / et neh 85th ty SE OIES 8 

Wenlttral eiOwis. 2 o<).rs o¢ic y. UBER SAS TE ek 3 ae lees) 8. Re 4 (one double). 

WAOESAlITOWsss ois aiee ee tee Se. sehr er rere ere 4 (2 rows, 2 each) 
Shape of tail of male..... Slopes gradually to a 

PON fete ee eee Bends sharply ventrad to a 


short, blunt end 


The comparison of the two species shows that the less common 
one agrees with Ascaris triquetra Schrank, which earlier writers 
believed to be synonymous with A. mystax Zeder and A. marginata 
Rudolphi. Marcus (1906) had identified his A. canis with the A. 
marginata studied by Kultschitzky (1888). Cytological examination 
of the sex cells of this Ascaris triquetra Schrank shows that there are 
twenty tetrad chromosomes, arranged in ten pairs, and also two 
unpaired *tetrads, as the diploid number. This agrees with the facts 
recorded by Marcus for his material, and I believe, therefore, that the 
Ascaris studied by him was also Ascaris triquetra Schrank, known to 
Kultschitzky as Ascaris marginata Rudolphi. 

My work, then, has shown that, while Ascaris canis Werner is the 
common parasitic nematode of the dog, Ascaris triquetra Schrank 
may be an inhabitant of the same dog that harbors individuals of the 
species A. canis Werner, though this occurs only rarely. It has also 
shown that the nematode studied by Marcus (1906) was probably 
Ascaris triquetra Schrank, rather than Ascaris canis Werner. 

I wish here to express my obligation to Dr. S. I. Kornhauser for 
his notes and especially to Dr. E. L. Mark for his supervision of the 
preparation of this paper. 


BIBLIOGRAPHY 


Edwards, C. L. 1911. The Sex-chromosomes in Ascaris felis. Arch. f. 
Zellforsch., 7: 309-313, Taf. 23-26. 

Glaue, H. 1908. Zur Unterscheidung von Ascaris canis und A. felis (Ascaris 
canis s. mystax). Zool. Anz., 33: 785-790, 3 figs. 

1909. Beitrage zu einer Monographie der Nematodenspecies, Ascaris felis 
und Ascaris canis. Zeit. f. wiss. Zool., 95: 551-593, 26 figs. 

1910. Beitrage zur Systematik der Nematoden. Zool. Anz., 35: 744-759, 5 figs. 

Kultschitzky, N. 1888. Ueber die Eireifung und die Befruchtungsvorgange 
bei Ascaris marginata. Arch. f. mikr. Anat., 32: 671-682, Taf. 26-27. 

Marcus, H. 1906. Ei- und Samenreife bei Ascaris canis. Arch. f. mikr. 
Anat., 68: 441-491, Taf. 29-30, 10 text figs. 


REVIEWS AND NOTES 


The staff of the Research Department at the Severance Union Medical 
College in Seoul, Korea, of which Dr. Ralph G. Mills is director, has under- 
taken a review for English readers of current periodicals in Japanese medical 
literature. This is printed every two months in the China Medical Journal, 
and also circulated separately. The publication is likely to be of great impor- 
tance to parasitologists because of the activity in Japan at present in the 
investigation of diseases caused by animal parasites which play a great role 
in that country. 


The first (?) number, dated 1916 and recently received, contains a review 
with illustrations of long articles on the development of the supposed last stage 
in the life history of Paragonimus by Nakagawa, on the first intermediate host 
of that parasite by the same author, and on an investigation of the Lungfluke 
in Korea by Kakami, in addition to numerous other items mostly pathological. 
The reviews are very well written and present valuable material not otherwise 
accessible to the American investigator. a 


It is with great sorrow that the JouRNAL announces the death of one of 
its collaborators, the distinguished German helminthologist, Max Lthe, Pro- 
fessor at the University of Konigsberg, who died of wounds received in the 
war. The death of Lithe is a great loss to science and the world. His con- 
tributions to the literature of parasitology embrace important and extensive 
studies on Protozoa, Trematoda, Cestoda, Nematoda, and Acanthocephala. It 
is hoped to print at an early date a Picea sketch of Professor Lthe 
accompanied by a portrait. 


Harvard University has issued the first formal announcement of the School 
of Tropical Medicine. Courses are open to graduates of recognized medical 
schools so that the work becomes a part of the Graduate School of Medicine. 
Dr. Richard P. Strong is Director of the School of Tropical Medicine and in 
the work announced are courses in protozoology, helminthology, and tropical 
entomology. 


The Journal of Parasitology 


Volume 3 : DECEMBER, 1916 Number 2 


THE EFFECTS OF RADIATION ON THE DEVELOPMENT 
OF TRICHINELEA SPIRALIS 


WITH RESPECT TO ITS APPLICATION TO THE TREATMENT OF 
CTHER PARASITIC DISEASES 


E. E. Tyzzer anp JAMES A. HoNneIJ 


Since radium has been shown by biological experiment to have a 
pronounced effect on the development of the germ cells of various 
species, the possibility of its utilization in the destruction or even in 
the emasculation of certain parasites for which there is at present no 
efficient remedy appears worthy of consideration. It was thought that 
radium might be employed to advantage in the treatment of cases of 
schistosomiasis in which the bladder is involved, several of which have 
been under the authors’ observation for a considerable period of time. 
Although this condition is of common occurrence in certain parts of 
the world and although it is frequently attended with serious compli- 
cations, up to the present time no successful form of treatment has 
been discovered. Since the inflammation in this disease is produced 
by the presence of the ova in the tissues and since the worms from 
which the latter are derived, are situated in close proximity to the 
mucous surface of the bladder, this mode of attack seemed to be 
especially appropriate. It did not appear justifiable, -however, to 
undertake the treatment of human cases without a certain amount of 
preliminary experimentation. 

While the use of Roentgen ray for the treatment of schistosomiasis 
has been suggested,’ there appears to be an advantage in the use of 
radium or its emanation in this disease, for the bladder wail in which 
the worms are situated may be radiated directly from its inner surface 
and rays of shorter wave length may be utilized than is possible with 
the Roentgen rays. According to Packard (quoted by Abbe, 1914) 
the beta rays are more effective than the gamma rays in retarding 
the development of certain species. 


1. The advisability of employing the x-ray therapeutically in this disease was 
discussed by Doctor R. Gonzales Rincones of Venezuela at the recent Pan- 
American Scientific Congress at Washington. 


, 


aia THE JOURNAL OF PARASITOLOGY 


In the following experiments radium emanation was employed as 
follows: 

1. To radiate from the outside the abdomen of rats previously fed 
with the cysts of Trichinella spiralis. 

2. To radiate muscle containing encysted larvae of this parasite. 

3. To radiate the worms directly during their development in the 
intestine by feeding minute glass tubes containing radium emanation. 
Radiation was accomplished in several ways and further details will 
be presented with the account of each experiment. 

Technic.—The effects of radiation on the parasite were judged by 
either the failure of the larvae to develop in the intestine of rats and 
mice or by abnormalities in their development. It was thus important 
to determine the number of worms present in the intestine and also 
to note any retardation in their differentiation or growth. In order to 
count the worms, the intestine of the animal to which the larvae had 
been fed was cut into pieces of from 3 to 4 cm. in length. These were 
each placed on an ordinary microscopic slide, opened with fine scissors, 
and by fixing one end of the piece with tweezers the mucosa was com- 
pletely stripped from the muscular wall by several light sweeping 
strokes with the edge of a scalpel. The material obtained, 1. e., mucosa 
and softer portions of the intestinal contents, was spread slightly, and 
then pressed gently beneath a large 22 by 40 mm. cover glass. Witha 
microscope equipped with a mechanical stage, all the worms in such 
preparations may be readily observed and counted. Since the material 
is flattened into a thin film the anatomy of the worms is clearly 
apparent so that an accurate enumeration of the sexes may readily be 
made. In the earlier experiments equal amounts of muscle taken from 
corresponding portions of the body of an infected animal were used 
for infecting the radiated and the control series, respectively. Since 
this procedure furnished only approximately equal dosage in the later 
experiments with mice, the cysts contained in strips of diaphragm 
were counted and an equal number fed to each of a series of animals. 

Observations made during the course of this study failed to confirm 
certain statements, which occur quite generally in standard works, con- 
cerning the anatomical distribution of Trichinella spiralis, the ratio of 
the sexes, and the span of life of the adult male and female of this 
species. 

Distribution in Rats and Mice—Trichinellae are said to mature in 
the duodenum and jejunum and it might be inferred that the adults 
are confined to the first portion of the intestine. In the course of the 


2. The authors are indebted to Doctor William Duane for collecting and 
measuring the radium emanation used and also for suggestions as to dosage, 
filtration, etc. 


TYZZER-HONEIJ—EFFECTS OF RADIATION 45 


following experiments the worms were comparatively rarely present 
in the first portion of the small intestine of rats and mice, but were 
found in great numbers throughout the remainder. They not infre- 
quently occur also in the cecum and colon of mice and occasionally 
in the large intestine of rats. It appears probable that the small size 
of these host species may account for the presence of the worms in the 
large intestine since no great extent of gut would have to be traversed 
before reaching the cecum. 

Sex Ratio—According to Staubli (1909), great discrepancies with 
respect to this point are found in the statements of different authors. 
Thus Leuckart reports the females as greatly in excess of the males, 
in one instance in a 10:1 or 20:1, and in another instance in a 6: 1 
ratio. Zenker calls attention to the difficulty in finding the males on 
account of their smaller size. Askanazy, on the other hand, finds the 
males greatly in excess in the intestinal contents, but this was thought 
to be due to the fact that the females burrow into the mucosa, while 
the males remain free. Ostertag claims that the males and females 
are originally present in equal numbers, but that the former after 
copulation diminish in number, so that after 10 to 14 days only females 
are present. Both sexes were observed by Pagenstecher 56 days after 
ingestion. Staubli notes great variation in the sex ratio in different 
cases with respect not only to the mature adults, but also to the 
encysted larvae the sex of which he is able to distinguish. He 1s 
unable to account for this lack of uniformity in the relative number 
of the sexes. 

In order to avoid error in estimating the relative number of males 
and females it is important to examine the material in such a way 
that none will be overlooked. The males, on account of their smaller 
size, are not so readily detected, except with the aid of a microscope. 
Thus in 100 worms picked out with the naked eye from a suspension 
of intestinal contents and mucosa, not a single male was found; 
whereas a count made with the microscope of samples of the same 
material showed 31 per cent males 

Counts of 100 or more worms from the intestines of four rats of 
the present series showed the percentage of males to vary from 31 to 
41 per cent seven or eight days after injestion. In a total of 446 
worms, 160, or 36 per cent, were males. An approximation of a 1:2 
ratio was thus found in these animals. Rats killed seventeen or 
eighteen days after ingestion of infected muscle showed practically 
the same ratio, although only few worms were found. It is of interest 
to note that in one rat in which a single male was found unaccom- 
panied by any females, numerous larvae were found in the striated 
muscles showing that this male had outlived one or more females 
which had been present. 


EXPLANATION OF PLATE 


Fig. 1—A cross section of male and female Schistosoma haematobium situ- 
ated in a distended vein at the juncture of the submucosa and muscular wall 
of the bladder. This vessel is evidently occluded by the inflammation which 
the worms’ presence has excited. The intestinal ceca of the female are dis- 
tended with deeply stained material to the right of which is the ovary. 


Fig. 2—Male and female S. haematobium in longitudinal section. In the 
upper portion of the sectioned worms to the right a row of eggs is visible in the 
tubular uterus of the female. The acetabulum of the male is apparent, directed 
inwardly near the anterior extremity of the worms at the left. These worms are 
situated in veins beneath the submucosa, in this instance 4 or 5 mm. from the sur- 
face of the mucosa. The inflammatory changes in the latter are apparently due to 
the presence of numerous ova, for these are found surrounded by collections 
of exudate with which they are evidently discharged during the contraction of 
the bladder. Many ova also fail to reach the surface but become imbedded in 
the tissue where they are eventually destroyed, the shells persisting as foreign 
bodies. 


PLATE 


igure | 


Figure 2 


TYZZER-HONEIJ—EFFECTS OF RADIATION 47 


In mice fed with relatively small numbers of encysted larvae and 
killed four, five, six, seven and nine days later, there is considerable 
variation in the sex ratio, evidently on account of the small numbers 
of adult worms present in each animal. Including trichinellae sub- 
jected to radiation, together with those of the control mice, there were 
516 counted, and 159, or 30.8 per cent of these were males. Con- 
sidering the non-radiated separately, there were 167, of which 42, or 
25 per cent, were males. There was no marked diminution in the 
number of males from the fourth to the seventh day, and the number 
counted on the ninth day is too small to be of significance. The sex 
ratio of one male to two females is thus also approximated for this 
parasite in the mouse. 

Disappearance of Adult Worms from the Intestine—Whereas the 
males are said to diminish in number after copulation, which is accom- 
plished by the second or third day after ingestion, it is stated that the 
females may persist for five weeks or longer. Cohnheim claims to 
have observed trichinellae in great numbers up to the seventh week; 
Kratz found them seventy-seven days, and Leuckart twelve weeks 
after ingestion. In experimental animals the embryos are said to be 
liberated from adults for five to seven weeks after the ingestion of 
trichinous meat (Fantham, Stephens, and Theobald, 1916). It would 
appear from the findings in the rats of the following experiments that 
the adult worms disappear much earlier than the above observations 
by various authors would indicate. The data collected are presented 
in the following table. 


TABLE OF RESULTS 


a ee 
Period | Number of, 
Rat of Adult | Males Females Remarks 
Infection Trichinae | 


| J F y 
Wild 4853A....... 19 days...... INONGicra crest lec eslncicacie calories ccees sae Only a portion of intestine 
examined ; 
Wild 4853B....... 19 days...... None: <2 |S so RSAE Se Gad Bedan Speers Only a portion of intestine 
| examined 
White 4965 A.....| 25 days...... Present....| Present....| Present.... 
White 6590....... (CE eeaane Numerous... 31.5% 68.5% 146 counted 
| 
White 6592.......) 17 days...... ONC...2.205- a ooSeneeree Oner es cncen Entire intestine examined 
White 5618....... WEOAY Boca cc's Estimated | 36% 64% 100 counted 
600 
White 5619....... CAG ENC Rear Estimated 87% 63% 100 counted 
800 
White 5620....... 8 Gayeeccs:.: Estimated | 41% 59% 100 counted 
White 5622....... 18 days...... None....... Ore: oh Scien saraaece Ten inches of small intes- 
| tine examined 
White 5623....... TS CaS... <0s Ones cas ONG is ii0:5 «oof neae re eran eer Ten inches of small intes- 
tine examined 
White 5624....... 18-days:. i223. Oe coe netlreeean ae dese s Two........| Ten inches of small] intes- 
tine examined 
Witte :6625.525.6. 18) days. 2. INGMGioG 2 ee [scoot nls aceite. sotto on a aeietes Ten inches of small intes- 
tine examined 
White 5626....... 18 days...... MEDIC isre © cia l> visks wreceiers hace leaittatetaats nor eff Ten inches of small intes- 


tine examined 


48 THE JOURNAL OF PARASITOLOGY 


Numerous larvae were found in the skeletal muscles of all the 
negative rats showing that the adult worms had been present in the 
intestine of each. In one rat adult worms of both sexes were still 
present twenty-five days after ingestion, but in general their paucity 
or absence is notable in the animals killed later than the sixteenth day. 
In the last group, each animal of which was fed an equal amount of 
trichinous muscle, the most striking differences are shown with respect 
to the number of worms present in the rats killed seven and eight days 
and in those killed eighteen days later. This would indicate that this. 
parasite’s span of life in the intestine of the rat is rarely over three 
weeks, although there may occasionally be individuals persisting longer. 


In the following experiments radium emanation was used. Since 
this substance is transformed at a known rate, the amount of radiant 
energy available is constantly diminishing (approximately one half 
in four days), so that the dosage is greatest at the beginning of the 
exposure. 


I. RADIATION OF THE ABDOMEN. FROM THE SURFACE 
OF THE BODY 


In considering the effects of radiation on Trichinella spiralis, both 
the failure to develop as determined by the number present and the 
retardation of development as indicated by the absence of worm- 
shaped embryos in the females seven days after feeding were taken 
into account. Although various degrees of maturity were met with 
at this time, the presence or absence of worm-shaped embryos could 
be readily determined and served as a useful, although arbitrary, 
index. Small and evidently poorly developed males were also met 
with, but since they furnish no prominent feature by which their stage 
of development could be judged, they are not in this respect taken 
into consideration. 

EXPERIMENT 1 


February 18, 1916. Equal amounts of muscle containing encysted larvae were 
fed to four healthy rats and two of these served as controls, while the other 
two were radiated from the surface of the abdomen. Radium emanation 
enclosed in capillary glass tubes with 0.1 mm. of steel, 1 mm. of silver and 
a layer of adhesive plaster for filtration was used. This applicator was moved 
each day to a new area on the abdominal wall, from the entire extent of 
which the fur had been removed. One rat, 5591, which was radiated in this 
manner with a tube of 8.8 millicurie strength from the second day following 
ingestion of trichina, died six days later; that is, seven days after infection. 
An additional tube containing 6.4 mc. was added five days after feeding and 
two days before death. Another rat, 5593, was radiated in a similar manner 
with a very weak tube (3 mc.) from the sixth to the tenth day and with a 
96 mc. tube from the tenth to the seventeenth day, when it was killed and the 
number and condition of intestinal trichinae determined. 


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50 THE JOURNAL OF PARASITOLGGY 


EXPERIMENT 2 


March 10, 1916. Ten normal rats were fed with equal amounts of trichinous 
muscle mixed with bread and milk. Radiation was commenced at once with 
Rat 5717, a tube of 11.8 mc. strength being used. This animal was found 
dead at the end of seven days. Another rat was radiated with a weak tube 
(2 mc.) from the eighth day and also with a stronger dose (15.5 mc.) from 
the eleventh day. It died 16 days after infection. The results obtained in 
both experiments are combined in the table on the preceding page. 


While these results failed to demonstrate that radiation is of 
therapeutic value in the treatment of trichiniasis, they indicate that it 
appreciably modifies the development of the parasite in the intestine. 
Radiation after the females have become ripe, that is after the sixth 
day, fails to affect an earlier disappearance of trichinellae, or to pro- 
duce distinguishable injury to them. In fact, the worms appear to 
persist longer and to be unusually large and well developed in the late 
radiated animals. That larvae had continued to be liberated from the 
female worms was shown by the presence of very small as well as 
partially developed worms in the striated muscles of these rats. 
Although the control rats killed seventeen and eighteen days after 
feeding on trichinous meat furnish few or no adult trichinellae in the 
intestine, numerous larvae were found in the skeletal muscles of all 
showing that infection had occurred. 

Early radiation apparently had a greater effect on the development 
of trichinellae in the intestine. Radiation of the rat’s abdomen from 
the second day after the ingestion of the cysts resulted in a retardation 
of development as shown by the number of immature females; 32 per 
cent of these showed no fully formed embryos as compared with 
2 per cent in the control animals. Females were observed in the 
radiated rat which were so backward in their development that they 
were considerably smaller than normal males, although seven days had 
elapsed since their ingestion. There appeared to have been no general 
failure of the immature females to become inseminated, although this 
may have been accomplished later than normally. In only a few small 
undeveloped females was the receptaculum seminis not filled with 
spermatozoa. Subsequent experiments have shown that under normal 
or ordinary conditions the worms almost without exception are fully 
developed seven days, and usually six days, after they have entered 
the alimentary tract. Only a few larvae were found on careful search 
in the diaphragm of the rat radiated from the second day, whereas 
they were present in great numbers in the diaphragm of the control 
rat. Still earlier radiation, that is, from the time of the ingestion of the 
encysted larvae, appears to be even more effective, and in the animal 
in which this was carried out only two females, neither of which con- 


TYZZER-HONEIJ—EFFECTS OF RADIATION 51 


tained embryos, were found. The three rats which served as controls. 
for this animal each showed numerous well developed worms estimated 
at 600, 800, and 1,000, respectively. 

Since the radiation employed was fatal to three of the four trich- 
inous rats, the possibility that the injury to the host might indirectly 
affect the life of the parasites may be considered. That changes in 
the host resulting from radiation do not tend to destroy the worms is 
shown in the late radiated rat in which adult worms persisted longer 
than in the controls. 


II. THE RADIATION OF ENCYSTED LARVAE BEFORE INGESTION 


It appeared important, in order to estimate the dosage appropriate 
for the employment of shorter radium rays, to radiate the larvae 
before they were fed to the animals. For this purpose the filtration 
through the millimeter of silver was dispensed with and capillary glass 
tubes of emanation enclosed in steel tubes having walls 0.1 mm. in 
thickness were employed. Under-estimation of the effects of the short 
rays necessitated repeating the experiment several times. In Experi- 
ment 3, which is not presented in tabular form, none of the larvae in 
meat radiated with a 5.9 me. tube for six and for three days developed 
when fed to mice. Control mice fed with untreated meat in every 
case showed trichinellae when killed later on. 


For the purpose of making the observations 1nore accurate, equal 
numbers of encysted larvae were fed to each animal in all subsequent 
experiments. The encysted larvae were radiated by wrapping strips of 
mouse diaphragm around the steel tube containing the emanation, in 
this way ensuring fairly uniform radiation of all portions of the 
muscle. The layer of muscle around the steel tube nowhere exceeded 
1.5 mm. in thickness. The strips of diaphragm were subdivided when 
necessary so that an equal number of cysts could be fed to each mouse. 
This was accomplished by placing the bits of diaphragm in the mouth 
of the animal and holding the latter until the material was swallowed. 


EXPERIMENT 4 


April 14, 1916. Twelve mice were employed in this experiment. Two of 
these served as control animals, being fed each with 40 trichina cysts. The 
other ten were each fed 40 cysts which had been radiated for different -periods 
with 5.5 mc. of emanation enclosed in a capillary glass tube and filtered through 
0.1 mm. of steel. Tissue radiated was nowhere more than 1.5 mm. in thick- 
ness. Six of the animals were killed four and five days after this feeding, and 
a count made of the number of Tr. spiralis present in the small and large 
intestine of each. The other six were allowed to live for a longer period and 
the muscles were then examined to determine whether infection had taken place. 


52 THE SIOURNAL. OF PARASITOLOGY 


TABLE OF RESULTS—EXPERIMENT 4 


Mature |Immature 
No. Larvae Radiated Each Fed | SSS ae 
Mouse with 5.5 Me. 40 Larvae killed fof 2 fof 2? | tal 
5699 Control untreated....... AMT a. oraaens April 200. ceer 3 | 10 0 1 14 
5700 Control untreated....... 2 a re April 20 sincere 0 10 0 0 10 
5691 3 DIS. ab 3d) Osenmtaec.. - APTI 14 «seater tole AprilecO rraieislels One. 0 0 0 
5692 3) DIS. Ab so iOreeecieits «3 /\y iy) | a Ce ee ASPYAEZO: errsterere:s 0 0 0 0 0 
5693 6 Dts ab os One ase so JN Wa We ae | Apprile20nemmaeae 0 0 0 0 0 
5694 GUNTSNiab SaIOstee. sec. PAjs3 pV WA Rares Aprill2iyjescsers 0 0 0 0 0 
| 

5695 12 res bho O soe a a0)! April 140 i2ee May 40 s0 deren es Muscles negative 0 
5696 Ae iy er 0350) Oy aah oogoda MATYIL V4 2. laverere HE ae Ree iarion Muscles negative 0 
5697 ZANTNs ebb toh Cuetec 3:5. < 7X 0101 3s i ae May Amanecer Muscles negative 0 
5698 Qa OATS ates WO ct cs osies:3 A/V 1c | Bel ee Mayra ctticmss Muscles negative 0 
5701 4S DISS elu ssude eran srsie'e +6 ADT Aas ceo aees Maya da. eer Muscles negative 0 
5702 48° hrs) ates #Oa.c....- Pele WADI 14%. .ni seed May 4.0% elo. es Muscles negative 0 


EXPERIMENT 5 


April 21, 1916. Fourteen mice were employed, two receiving untreated cysts, 
the others receiving equal numbers of cysts which had been radiated for dif- 
ferent periods of time at room temperature. The technic employed was the 
same as that in the preceding experiment but shorter exposures were made. 
The radiated tissue was nowhere more than 1.2 mm. in thickness. The mice 
were all killed five or six days after the feeding. 


TABLE OF RESULTS—EXPH#RIMENT 5 


Mature |Immature 

No. Larvae Radiated | Each Fed : O-— 
Mouse with 7.1 Me. | 40 Larvae Killed fof Q fof 2 | tal 

a ft a | a | J | 
5738 Control untreated........| April 21........ PANT eG neste isiots 0 3 0 q 4 
57389 Control untreated........| April 21........ ATCA ieee 2 8 0 0 | 10 
5726 DAG IMITTGES pa aisle.s « «\s\s,0. 210) | Apriligte ne. ApTUZ6. se sea 1 1 2 8 | 12 
5727 246 MinuteS....--....0+-- | April 21.2 ose. VAST Zities erereretars 6 | 10 0 0. "16 
5728 & minmubesias...... J... April oi. 25 2 April 26........ gi) oe 82h Sees 
5729 Spits have i3s)515 oD UB COORE One ATI eects as ADIL: wis cieetels TE Wy oc 0 6 | 10 
5730 10 THINMTOs eect c.s ec cice Arisol eee ere | April 26;..u.02% 1 8 2 4 Walp 
5731 AO MMIDUWLER paresis. «s+ 52 sie > APTA Pace APTI 27 /ereista,nats 0 0 0 0 0 
5732 ZOMMINUCEA Eis: a:2015 6se.e April 21 25S) Aprill 26. 06ese 6 0}; 0 0 0 0 
5733 20 UIUC eee ie os's 5x00, ADT 21 ae ADIN Zi acct if Pa 0 Oo | 19 
5734 AQUMINMILER Tee ce cisscs0e 80s Jy yall Al ooscac 3 AD raO26 sacra ter 0 0 0 0 0 
5735 AQ THINICOS ites o\ccieceeces ADT Qh viecaer ADIL 2ipemaeee 1 1 0 0 2 
5736 SO WIM ETE Saisie esse, oe0ie0.e AMTWUDI Ey. occ April! 265 5 ccmace 0 0 0 0 0 
5737 SOT GER R eet cisiaicinarecis.e’s ADU Zi eee APTI iene eter 3 0 0 0 0 0 


It is apparent from the above experiment that radiation for eighty 
minutes with 7.1 mc. is fatal to the encysted larvae, but the results 
obtained for the next shorter periods are rather variable, one positive 
and one negative result being obtained in each of the three successive 
periods. Radiation for two and one-half and for five minutes appears 
not to have been markedly injurious. It was thought possible that since 
certain portions in the length of the steel tube employed were less 


TYZZER-HONEIJ—EFFECTS OF RADIATION 53 


radioactive than others, ceftain portions of the diaphragm exposed 
may have been subjected to less radiation accounting for the irregu- 
larity of the results obtained. On account of this it was considered 
necessary to repeat this experiment, paying especial attention to the 
equal radiation of all parts of the material used. 


EXPERIMENT 6 


May 1, 1916. Ten mice were employed, two served as controls, and the 
others received equal numbers of encysted larvae radiated for various periods. 
These were killed four and five days after this feeding and the intestine 
examined for Tr. spiralis. , 


TABLE OF RESULTS—FEXPERIMENT 6 


| 
| Mature |Immature 
No. Larvae Radiated | Each Fed —_—|—_—_-_——| To- 
Mouse | with 6.6 Me. | 40 Larvae Killed rol Q of Q | tal 
5 eS ee (es se eS a 
5749 Contre] untreated........ 1 Ie (es i AVL DO Oita s/o ainine « 4 0 0 1 5 
5750 | Control untreated........ | May 1.......... We fe aerate 1 3 0 8} 12 
5747 MOOMminuteSs.ecccott sented WAY Tera mates MAN csicgis wince 1 0 2 5 8 
5748 PU MMULERR he pcs cae te ore ay Dretseitucss ate AERCUG Sc oss sian « 0 0 0 1 1 
745 By TRENUILER a6 of): k's oss 6 noe May ds. s.ccacne MSE Diels clseccc0:s5 0 0 0 0 0 
5746 SOA TRITIEER ost ssseerelame sais MG Ye Done sio.2;0'0;« MOG oes. 3. ss 0 0 0 0 0 
5743 AY THINNER ee) aselo seo «eed Mayle sSvsc cece Motch o sis:s. 3's 0 0 0 0 
5744 PAO NII EOS oio6 oc .arne tis viele A Sl eae SDE Mave Gee. < ~.:. - 0 0 | 0 0 0 
Salis s LOOMMINIER see «20-6 ocioeie ee May draiccstece Mey ibise:.....: OO OL eoe io 
5742 HOO TMINULCR ace acco sseree Marr aie se aa iS 0 0 0 0 0 
| 


From the three preceding experiments the lethal dosage of radia- 
tion for encysted trichinae is determined. They are made non- 
infectious for mice by radiation with 6.6 me. filtered through thin 
glass and 0.1 mm. of steel in an exposure of thirty minutes. The 
cysts exposed were at a distance of not over 1.5 mm. from the source 
of radiation. It would be of interest to learn more concerning the 
effects of this amount of radiation on encysted larvae whether they 
are killed outright, or the cyst made more resistant to the digestive 
juices or the larvae injured to such an extent that they are passed from 
the alimentary tract before they can recover sufficiently to maintain 
their existence. In all of the present experiments only the immediate 
result of radiation as shown by the absence or by the arrested develop- 
ment of worms, has been determined. It would be of considerable 
interest to note whether any remote or late changes are brought about, 
but this probably would be more readily determined in a free-living 
rather than in a parasitic species. 


lil. THE DIRECT RADIATION OF TRICHINELLA SPIRALIS FROM THE 
INTERIOR OF THE ALIMENTARY TRACT 


Since the short rays were found to be effective in the destruction 
of the larvae the direct radiation of the worms from the interior of 
the intestine was next undertaken. Through radiating the interior of 


54 THE JOURNAL OF PARASITOLOGY 


the intestine by means of tubes of emanation fed to the animal it was 
hoped to utilize very short rays. The movement of the intestinal con- 
tents was expected to prevent undue burning of the mucosa of the 
small intestine and the incorparation of the tube in more or less solid 
fecal material was hoped to protect the wall of the large intestine from 
serious injury. Minute tubes of emanation were prepared by Doctor 
Duane. Since these measured only from 2 to 3 mm. in length and a 
fraction of a millimeter in diameter, they could be readily introduced 
into the stomach of the mouse. In order to accomplish this, a large 
syringe needle, the point of which had been ground off square, covered 
with paraffin and dipped in oil so that it could be readily passed down 
the esophagus of the mouse, was used. The emanation tube having 
been placed in the needle, it was forced by a plunger into the stomach 
of the mouse. It could readily be determined at any time whether the 
tube had been passed from the intestine or was still in the body of the 
mouse by placing the latter on the ionization chamber of the measuring 
apparatus. 
EXPERIMENT 7 


June 27, 1916. Nine mice were each fed 40 trichina cysts in bits of mouse 
diaphragm. Minute glass tubes of emanation were introduced into the stomach 
of three of these, one receiving a tube on the first day, another on the second 
day, and another on the third day. 


TABLE OF RESULTS—EXPERIMENT 7 


No. No. Fed i Radium Fed Died | No. Tr. Spiralis 


| 

5803 80 cysts........ 3.1 me. ¢ hours later.....:..... Killed 5 days....| 24 (4 immature) 
5804 SOMGYBESs < <ssfenie 4.1 me. 28 hours later........... Dead 6 days.....| 57 

5805 BUMEV RUS =< c.tan ts 3.2 me. 51.5 hours later......... | Dead 6 days..... | 46 

5806 SO cysts)... ..'. MIGHUPOL, wiser coltasle cee ete wine ae Killed 5 days....| 33 

5807 SO CYSUBS. 5 c.0i MS OTEGTONE cieracincctoeic oa neisereciacte'e Not examined 

5808 SO CYBEGS ios. MDNETON SE cacaccedes aemene seb ede Not examined 

5809 80 Cysts: ....... Wontrol, oe cpenae aus acoumercercnae Not examined 

5810 CO CURDS os paces WOnTols S56 co. ceaec seen nolo Not examined 

5811 80 cysts... ..... MIOUELOL crates inaves cece saanane Not examined 


— 


In the first mouse (5803) the tube failed to pass from the intestine 
during the four days and fourteen hours which elapsed before it was 
killed on account of its weak appearance. The tube fed the second 
mouse (5804) was still in its body after three days, but had been passed 
when found dead two days later. The tube fed the third animal (5805) 
was passed within forty-eight hours. Notwithstanding the small dosage, 
3.2 me., for so short a period, this animal died before the end of four 
days after its introduction. This mouse presented a reddened area in 
the wall of the large intestine. The others showed no local effects of 
the radiation, but all showed a striking shrinkage in the size of the 
spleen so characteristic of radiated animals. These results show unmis- 
takably that in the mouse even fatal doses of radium emanation acting 


TYZZER-HONEIJ—EFFECTS OF RADIATION 55 


from the interior of the intestine fail to prevent the development of 
Tr. spiralis. It is quite apparent that this parasite is not especially 
vulnerable to rays which have an immediate destructive influence on 
the lymphoid tissue of the host. 

The results of the foregoing experiments are thus rather discourag- 
ing with respect to the application of radiation to parasitic worms. It 
is rather remarkable that Tr. spiralis is so little affected after com- 
mencing its development within the body when it, in its encysted state, 
is so quickly destroyed by radiation outside the body. Whether internal 
radiation would accomplish more in larger, more resistant animals 
remains to be determined. The acceleration of the passage of radium 
emanation through the large intestine by the employment of a cathartic 
might be of value, but was not tried in these experiments. It would 
appear, however, that, under the conditions of the above experiment, 
radiation destroys the resistance of the animals before the parasite is 
markedly affected. 

These results do not, therefore, furnish an experimental basis for 
the treatment of schistosomiasis by radiation from the interior of the 
bladder. Although this treatment would for some reasons appear 
especially applicable to this disease, it would probably be impossible, 
with the short rays, to reach all the adult worms, for they are fre- 
quently situated at a considerable distance from the surface of the 
mucosa. Treatment of this disease by radiation, although justified on 
theoretical grounds, must for the present be regarded as experimental 
in character, and, even if no untoward results occur, it will be difficult 
to determine just what has been accomplished. On the other hand, 
the localization of the worms and the relatively large size of the host 
are distinctly in favor of this form of treatment. The fact that it is 
a disease frequently attended with serious complications would appear 
to warrant cautious treatment by radiation, provided that the experi- 
mental nature of the treatment is explained to the patient, and provided 
that changes in the local condition may be followed by cystoscopic 
examination and by observations with reference to the number of ova 
discharged in the urine. 

SUMMARY 


By radium radiation from the surface of the abdomen of the rat 
the injury of fully developed Trichinella spiralis has not been accom- 
plished. These worms appeared well developed and persisted longer 
than in controls. . 

Similar treatment from the second day after the ingestion of cysts 
has apparently resulted in a retardation of development, 30 per cent 
of the females being immature. 


56 THE. JOURNAL OF PARASITOLOGY. 


In a rat radiated in this manner from the time it was fed trichinous 
meat, only two immature worms were found seven days later, indicat- 
ing that radiation of the larvae before they have entered upon their 
period of development free in the intestine is fatal to them. 

The radiation of encysted larvae with 6.6 me. of emanation, 
0.1 mm. steel filtration, at a distance of not over 1.5 mm. for thirty 
minutes renders them non-infectious for mice. 

Radiation from the interior of the intestine—employing emanation 
in minute glass tubes for the utilization of short rays—in amounts suffi- 
cient to cause the death of the mice employed has not prevented the 
development of Tr. spiralis. 

These results fail to furnish an experimental basis for the treat- 
ment of schistosomiasis by radiation, so that if the latter is attempted 
it should be regarded as an experiment rather than an approved mode 
of procedure. 

Observations on the life history of Trichinella spiralis made in the 
course of these experiments indicate that certain points emphasized in 
books of reference do not apply to the development of this parasite 
in rats and mice. 


Trichinella spiralis is found only in small numbers in the duodenum 
and jejunum of rats and mice which show great numbers in the lower 
portion of the small intestine. It is also occasionally found in the 
cecum and large intestine. 

The life of this parasite is comparatively short in the rat, and it is 
found to have disappeared or is present only in small numbers eighteen 
days after infection. 

No evidence has been obtained that the males disappear early in the 
infection. A sex ratio of 1 ¢ : 2 2 observed six and seven days after 
infection has shown no marked change for the ten days following. A 
male Trichinella has been found in a rat from which all females had 
diseappeared. 

REFERENCES 


Abbe, R. 1914. Radium Beta Rays. The Efficient Factor in Repressive 
Action on Vital Cells. Medical Record, Nov. 28, 1914. 

Staubli, C. 1909. Trichinosis. J. F. Bergmann, Wiesbaden. 

Fantham, H. B., Stephens, J. W. W., and Theobald, F. V. 1916. The 
Animal Parasites of Man. Wm. Wood and Co., New York. 


NOTES ON SOME NEMATODES FROM 
FRESH-WATER FISHES * 


Henry B. WarD AND THomMas B. MacatTu 


The parasitic nematodes are of conspicuous importance in the field 
of human disease and also in diseases of the domestic animals, and in 
his treatise on fish diseases, Hofer (1906), discussing the significance 
in fish culture of parasites and parasitic diseases, states that among 
them the nematodes outrank all others in number of types. Yet as 
fish parasites these forms are almost unknown in North America, and 
references to them are confined to a few brief notes, almost all of 
which came from the pen of the distinguished Philadelphia micro- 
scopist, anatomist, and parasitologist, Joseph Leidy, whose pioneer 
work published between 1850 and 1886 includes many records of 
great value on this group. 

In this little-explored field the senior author has been making 
observations for many years and in collaboration with the junior 
author was led recently to undertake an extended study of nematode 
parasites from North American fresh-water fishes which has yielded a 
number of new and interesting forms; these are briefly described here 
in advance of the appearance of the complete article in which will be 
given fuller data on the structure and relationships of these species. 
Especial thanks are due the United States Bureau of Fisheries for aid 
in securing material. 

It is interesting to note that among the eight forms described as 
new species, three fall within new genera and five agree sufficiently 
with European forms to be listed in already existing genera. Seven 
out of the nine forms described in this paper come within the limits 
of the Spiruroidea, so that this superfamily appears to hold a prominent 
place among parasites of fresh-water fishes. 

Cystidicola Fischer von Waldheim 1797.— The type species 
C. farionis from the air-bladder of the trout and other fishes is a com- 
mon form in Europe; it has also been reported from the lake trout of 
Lake Erie. Leidy (1886) described a parasite of the air-bladder of 
the lake trout as Filaria stigmatura. We have the same parasite from 
the white fish, lake trout, and lake herring in Lake Erie, Lake St. Clair 
and Lake Michigan. It is clearly not a Filaria, but belongs to the 
genus Cystidicola, and should be called C. stigmatura. Two small 


* Contributions from the Zoological Laboratory of the University of IIli- 
nois, No. 78. 


58 THE JOURNAL OF PARASITOLOGY 


uncinate lips, a short buccal capsule or tubular pharynx followed by a 
long esophagus divided into an anterior muscular and a_ posterior 
glandular region, indicate the position of this form in the superfamily 
Spiruroidea created by Railliet and Henry (1916) and in the family 
Camallanidae established by the same authors. 

Camallanus Railliet and Henry 1915.— Among the commonest 
and best known parasites of fresh-water fish in Europe is the so-called 
Hooded Worm, easily recognized by its horny oral armature in the 
form of heavy ribbed valve-shaped lips of dark brown material with 
corner anchors of trident form. More than fifty names have been 
given these worms which most often have been assigned to the genera 
Dacnitis and Cucullanus, but have recently been definitely located in a 
new genus, Camallanus, by Railliet and Henry (1915a). Though in 
habit hookworms by virtue of their persistent attachment to the intes- 
tinal wall, they do not possess a buccal capsule like all true hook- 
worms; the valve-shaped lips form powerful lateral jaws that grasp 
the tissue, whereas the true hookworms possess a hollow cup into 
which the tissue is drawn by suction. Two species have been found in 
North American fresh-water fishes. The first is 

Camallanus ancylodirus noy. spec.— The head (Fig. 4) is bent 
sharply ventrad, whence the specific name. The mature female is 
25 mm. long by 0.56 mm. in maximum diameter. The caudal tip is 
bluntly conical, and 0.45 mm. in front of it lies the anus. The vulva 
is three-fifths of the length from the anterior end. The trident in the 
oral armature has three or rarely four prongs, irregular in form and 
0.21 mm. long. The lips measure 0.142 to 0.168 mm. long by 0.18 to 
0.187 mm. broad. The uterus, loaded with minute embryos, fills the 
entire body save at the extreme ends. The anterior region of the 
esophagus is 1.416 mm. long and 0.096 to 0.15 mm. broad; the posterior 
region measures 1.368 by 0.072 mm. 

Mature males measure 15 mm. in length by 0.38 mm. in breadth 
at the center of the body. The lips are 0.126 mm. long by 0.12 mm. 
wide, and the trident arms 0.18 mm. long. The anus (Fig. 10) lies 
0.156 mm. from the extreme tip. The caudal alae are 0.92 mm. long. 
The two spicules (Fig. 11) are nearly equal in length, but one is only 
half as heavy as the other. The posterior end is rolled in a half circle 
and the number of the caudal papillae could not be determined. 

This parasite came from the intestine of the German carp at 
Fairport, Iowa, and is the first reported from that host on this conti- 
nent. Evidently it is a native species that has accommodated itself to 
this new host, and its original host is as yet unknown. Instances are 
infrequent in which a host is known to have acquired an entirely new 
parasite. 


WARD-MAGATH—NEMATODES FROM FRESH-WATER FISHES "39 


Camallanus oxycephalus nov. spec.— Only females were found. 
Length up to 25 mm. and maximum breadth 0.27 mm. The anterior 
end (Fig. 3) is perfectly straight and much smaller than the preceding 
species. The body tapers regularly from the center about equally in 
both directions. A small but distinct constriction, or “neck,” lies just 
behind the oral tridents and conforms to the curvature of the trident 
branches. The muscular esophagus is 0.47 mm. long and 0.085 mm. 
in diameter at the narrowest point, expanding near the end to 
0.105 mm.; the second region of the esophagus is 0.57 mm. long, of 
nearly equal diameter throughout and as broad as the maximum 
breadth of the muscular esophagus. The vulva is located at the 
anterior margin of the middle third of the body. This species was 
taken from the intestine of the white bass and of the black crappie at 
Fairport, Iowa. 

Cucullanus O. F. Miller 1777.— As Railliet and Henry have 
shown recently (1915a), the forms which rightly belong here are 
those to which the name Dacnitis Duj. 1845 has often been applied. 
They are characterized as follows: 

Anterior end bent dorsad. Mouth elliptical with major axis dorso- 
ventral, bounded by two lateral valves recalling those of Camallanus. 
Esophagus pestle-shaped without bulb. Males without caudal alae ; 
two equal spicules and an accessory piece. Preanal sucker without 
chitinous ring. Female with vulva near center. In alimentary canal 
of fish. 

The genus was placed in the Heterakidae by Railliet and Henry 
chiefly because of the preanal sucker of the male. However, a ventral 
sucker is by no means confined to this family, and this one has no 
marginal ring such as is present in other Heterakidae ; finally, the three 
lips of the Heterakidae are wanting. Accordingly, we have placed the 
genus in the Spiruroidea with which it agrees in the lateral valves at 
the mouth and the double esophagus, both characteristic of that group. 
The genus is the sole known representative of a family, the Cucul- 
lanidae, which differs from all other forms in the Spiruroidea through 
the possession of the ventral sucker in the male. 

Cucullanus clitellarius noy. spec—Body generally uniform in diam- 
eter, except for clitellar-like swelling 1.5 mm. from anterior tip (Fig. 
5). Head bent dorsad 60 to 90 degrees. On each oral margin three 
papillae. Oral valves 0.45 by 0.32 mm. in female, and 0.33 by 0.24 mm. 
in male. 

Males 10 to 11 mm. long by 0.38 mm. broad. Esophagus 1.45 mm. 
long and 0.12 to 0.22 mm. wide. Caudal region (Fig. 9) bent in a 
single turn. Ventral sucker 0.51 mm. anterior to anus, 0.1 mm. in 
diameter. From anus to tip of body 0.39 mm. Spicules 1.62 mm. long, 


60 THE JOURNAL OF PARASITOLOGY 


0.035 mm. broad, shaped like a gouge; accessory piece dagger-shaped, 
0.06 long by 0.015 mm. broad. Two small papillae just in front of 
anus; four pairs of postanal papillae, of which two pairs are large, 
rounded, and only 0.012 mm. from extreme tip. 

Females 12 to 17 mm. long, 0.5 mm. broad. Esophagus 1.6 mm. 
long and 0.13 to 0.32 mm. wide. Distance from anterior end to vulva 
from five-ninths to two-thirds total length. Uterus and ovary double. 
Ova 63 by 46n. 

Parasitic in intestine of lake sturgeon (Acipenser rubicundus) in 
Lake St. Clair. 

This genus has been heretofore the sole representative of its 
family, but among parasites in fresh-water fishes we have met another 
type that is sufficiently related to fall within the limits of the family, 
and yet cannot be brought within the limits of the same genus. To 
receive it a new genus has accordingly been created with the following 
characteristics : 

Dacnitoides nov. gen—Much like Cucullanus, except that head is 
not flexed and body is straight; spicules lack accessory piece, and only 
a single ovary is developed. A well developed intestinal cecum is 
present. ; 

Dacmitoides cotylophora nov. spec——Males 4 to 6 mm. long and 
0.2 mm. broad. Mouth dorso-ventral, bounded by lateral valves. 
Cuticular ridge with three papillae at extreme anterior margin of each 
valve. Esophagus 0.5 to 0.6 mm. long, 0.06 to 0.12 mm. broad, dis- 
tinctly divided into two regions; anterior region 0.2 mm. long. Intes- 
tine large, provided with dorsal cecum extending anteriad to junction 
between two regions of esophagus. Ventral sucker (Fig. 7) 0.41 mm. 
in front of anus, which is 0.12 mm. from posterior tip. Spicules 
0.89 mm. long and only 5 broad. Caudal papillae: one pair on ante- 
rior margin of sucker, four pairs between sucker and anus, four pairs 
of postanal papillae and one single median papilla immediately in front 
of anus. 

Females 4 to 5.5 mm. long by 0.28 mm. in width at vulva; body dis- 
tinctly short and heavy. Anterior end (Fig. 6) rounded, posterior end 
acutely pointed. Anus 0.14 mm. from posterior tip, with four slender 
spines halfway between. Vulva about five-eighths of total length from 
anterior end. Anterior and posterior uterine branches, but latter ter- 
minates blindly, and only former has an ovarian tube at end. Eggs 
measure 65 by 40p, and contain embryos in early clearage stages. 

This parasite was common at Lake St. Clair in the intestine of the 
yellow perch (Perca flavescens), and more rarely of the wall-eye 
(Stizostedion vitreum). 

In that this species possesses an intestinal cecum it resembles the 
family of the Heterocheilidae which Railliet and Henry established 


WARD-MAGATH—NEMATODES FROM: FRESH-WATER FISHES 61 


among the Ascaroidea om the basis of the development of such an 
organ. In other anatomical features it departs as widely from that 
family as it does from the Heterakidae. 

An interesting parasite was taken from the intestine of the bowfin, 
or fresh-water dog-fish, Ama calva, both in Lake St. Clair, Michigan, 
and at Fairport, Iowa. It is of a very generalized character and hence 
difficult to define except in negative characters, or to locate in the sys- 
tem. While we are inclined to place it in the Spiruroidea under the 
family Spiruridae, we must confess that this decision is open to criti- 
cism, and it may have to be made the representative of a new family 
standing half way between the Ascaroidea and the Spiruroidea. It 
must certainly be made a new genus characterized as follows. 

Haplonema nov. gen.—Body rather robust, but not large; anterior 
end bent or coiled, without lips or papillae, but with lateral alae 
(“wings”). No buccal capsule; esophagus muscular, without bulb, 
divided into two regions by partition near center. Posterior end of 
female straight, or slightly curved behind anus, with two minute 
papillae. Posterior end of male without bursa or alae, with two equal 
spicules of moderate length. Two pairs of preanal papillae and three 
pairs of postanals. Ovary laid in transverse loops ventral to intestine 
in both anterior and posterior regions. Uterus with anterior and pos- 
terior branches, vulva near center of body. Oviparous. 


Haplonema immutatum noy. spec.— Males iess frequent than 
females, measure about 9.5 mm. in length by 0.2 mm. in breadth 
anteriorly and 0.18 mm. posteriorly. Females about 15 mm. long by 
0.31 mm. in maximum breadth in the anterior region. Vulva five- 
eighths of length from anterior tip. 

Anterior end (Fig. 1) bent in a half circle with lateral cuticular 
folds extending back 2.5 or 3 mm. from anterior tip. Esophagus 
prominent, muscular, 0.65 mm. long in male and 0.8 mm. in female; 
width of esophagus, 0.06 mm. anteriorly and 0.1 mm. near its posterior 
end. Esophageal partition inconspicuous, near center of length; two 
regions alike in structure. 

Spicules (Fig. 2) two, equal, flat, ribbon-like, 0.75 mm. long, by 
0.02 mm. wide. Eggs abundant, with moderately thick, smooth shells ; 
average size 65 by 45y. 

The genus Spinitectus was established by Fourment in 1883 to con- 
tain parasites of fishes characterized by circles of retrorse spines on 
the posterior margins of transverse rings. To the four species known 
in Europe we add a new form common in some places here. 

Spinitectus gracilis noy. spec-—Mature females 11 to 19 mm. long; 
body divided more or less distinctly into slender transparent anterior 
region about 6 mm. long and 0.066 mm. wide, and larger, darker, more 


62 THE JOURNAL OF PARASITOLOGY 


opaque posterior region about 12 mm. long and 0.14 mm. broad, 
crowded with internal organs, especially the uterus gorged with eggs. 
Tail abruptly conical, 0.096 mm. from anus to tip in female, only 
0.066 in male. Vulva three-fourths length of body from anterior end, 
inconspicuous. 

Spinous rings begin 0.12 mm. from anterior tip. First 6 to 8 rows 
more prominent than those later. Subsequent rows (up to 28 or 30) 
smaller, closer, with spines much lighter and shorter; last rows difficult 
to detect. Total about 130 rows. Rings about 0.03 mm. apart at 
anterior end, and contain 40 to.50 spines in each circle. Largest spines 
Su long, smallest less than half as long. 

Male 12 mm. long by 0.042 mm. in diameter in anterior region and 
0.75 mm. at widest point. Posterior end coiled in spiral with 2 to 3 
turns. Narrow lateral alae 0.33 mm. long, near caudal tip, not sup- 
ported by papillae or ribs. On ventral surface, 1 mm. anterior to anus 
a series of 4 to 8 longitudinal rows of small ridges, each about 5y long 
and 3u high. Spicules very heavy, longer scoop-shaped; shorter, 
arcuate, oblique, probably not protrusible. 

Pharynx tubular or funnel-shaped, short. Muscular esophagus 
narrow, 0.33 mm. long in female and 0.25 mm. in male, no marked 
boundary between it and glandular region. Egg-filled uterus large. 
Ova 41 by 24», with thick transparent wall and without polar processes. 

Some specimens 4 to 5 mm. long by 0.35 mm. broad have spines 
extending even to the anal region, 175 rows in all. From the vulva a 
broad vagina projects anteriad bearing at its inner end anterior and 
posterior uterine branches. 

This parasite occurs in the alimentary canal of the black crappie, 
sheepshead, and white bass at Fairport, Iowa. In life it is transparent, 
and the spinous rings are very distinct. The worms are not attached 
to the wall, but lie free in the lumen of the gut. The spines are 
encased in masses of food particles. 

Railliet and Henry include in this genus as Spinitectus cristatus a 
form described by Linton from the hake as Filaria serrata. They use 
his description of the male for the genus since Fourment found no 
males in his material. Our form differs from Linton’s in the absence 
here of papillae he described and in the presence here of caudal alae he 
neither mentions nor figures. 

A nematode parasitic in the perch in Lake St. Clair is assigned 
‘without hesitation to the genus Ichthyonema. The genus is closely 
related to Dracunculus medinensis of man. In Europe various species 
are abundant, and widely distributed, but one has never been reported 
before this in North America. Possibly some of the worms listed as 
“Filaria” from American fishes really belong here. The species we 
have does not agree with any known form and is designated as 


WARD-MAGATH—NEMATODES FROM FRESH-WATER FISHES 63 


Ichthyonema cylindratewm nov. spec. — Male unknown, probably 
minute. Mature female 100 mm. long, of nearly equal diameter 
(0.48 mm.) everywhere. No lips or papillae. Esophagus 1.09 mm. 
long, 0.066 mm. in diameter. Vulva and vagina atrophied, no vestiges 
discernible. Uterus crowded with undeveloped ova (i. e., female 
unimpregnated ), ova almost spherical, measure 0.044 mm. in diameter. 

In abdominal cavity of Perca flavescens, Lake St. Clair. 

The worm was delicate, semitransparent, and very fragile owing to 
the thin body wall. The lateral lines are broad, light colored, and 
conspicuous. In Europe almost half of the females found are like our 
material, unimpregnated owing apparently to scarcity of males. This 
species is of great interest from its relationship to the Guinea Worm 
of man. 

Among the Ascaridae, Railliet and Henry(1915)have grouped thdse 
forms having either an intestinal or an esophageal cecum into a single 
family, the Heterocheilidae. One form we have studied falls within 
this group, but cannot be placed in any of the genera found in it, hence 
a new genus is created to contain it characterized as follows: 

Hysterothylacitum noy. gen.—Body without anterior tunic, but with 
narrow lateral alae (“wings’’). Lips three, not prominent. Esophagus 
long, slender, with terminal spherical bulb. Intestine with short simple 
cecum, arising from anterior end of intestine, directed posteriad. Males 
with two equal curved spicules, papillae(?). Females unknown. 

Hysterothylacium brachyurum nov. spec——Length of male 32 mm., 
maximum width just behind center of body, 0.66 mm. Lateral fin 
(Fig. 8) from base of lip to esophageal bulb or further; width one 
quarter the diameter of body. 

Esophagus 3.1 mm. long, 0.1 to 0.13 mm. wide; bulb with three 
teeth, cecum 0.94 mm. long, 0.08 mm. wide. Spicules equal, 0.72 mm. 
long by 0.045 mm. wide. Pyriform sperm-vesicle prominent. In 
stomach of black bass, Lake St. Clair, Michigan. 


REFERENCES CITED 


Fourment, L. 1884. Note sur un Nématode nouveau parasite du Merlan. 
Agin. “sci. nat. .zook, (6) “17; acts.5e78:pp., lpi: 

Hofer, B. 1906. Handbuch der Fischkrankheiten. Stuttgart. 2. Aufl. 359 
pp., 18 pl., 222 text figs. 

Leidy, J. 1886. Notices of Nematoid Worms. Proc. Acad. Nat. Sci., Phila., 
38 : 308-313. : 

Railliet, A., and Henry; A. 1914. Essai de Classification des “Heterakidae.” 
IX. Congr. Int. Zool., Monaco, pp. 674-682. 

1915. Sur les Nématodes du genre Goeszia Zeder. Bull. Soc. Path. exot., 
8: 273-275. 

1915a. Sur les Nématodes du genre Camallanus Raill. et Henry, 1915 
(Cucullanus Auct., non Mueller, 1777). Bull. Soc. Path. exot., 8: 446-452. 

1916. La famille des Thelaziidae. Jour. Parasitol., 2: 99-105. 


EXPLANATION OF PLATE 


Fig. 1—Haplonema immutatum; head of female showing lateral fin fold, 
excretory pore, two regions of esophagus, esophageal valve, and intestine. X 72. 


Fig. 2—Haplonema immutatum,; tail of male showing spicules and papillae. 


x 72. 


Fig. 3—Camallanus oxycephalus; head of female showing valves, trident, 
ring, and anterior region of esophagus. 72. 


Fig. 4—Camallanus ancylodirus; head of female showing oral armature and 
two regions of esophagus. 22. 


Fig. 5—Cucullanus clitellarius; head of female showing valves, double esopha- 
gus, esophageal valve and intestine; also half of clitellar-like swelling. 20. 


Fig. 6—Dacnitoides cotylophora; head of female, showing oral armature, 
esophageal regions, intestine, cecum, and anterior coils of ovary. 70. 


Fig. 7—Dacnitoides cotylophora; tail of male, showing sucker, spicules and 
papillae. > 70. 


Fig. 8—Hysterothylacium brachyurum; head of male showing lips, lateral 
fin-fold, esophagus, esophageal bulb, intestine, and cecum. 22. 


Fig. 9—Cucullanus clitellarius; tail of male, showing sucker, spicules and 
papillae. > 22. 


Fig. 10.—Camallanus ancylodirus, tail of male showing spicules and papillae. 


< 70: 
Fig. 11—Camallanus ancylodirus, spicules of male.  X 110. 


All reference lines in millimeters or tenths as indicated on plate. 


PLATE 


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ek eee 
Nate es 
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~ oe is 
eee 


T i. . ‘ a ra 
AC - ‘ 
iu a ae x 4 
on -) j = : 
va qn ios 
er, ) SS Oe a in 
tas She he! 
an =e OE te > 


o~ 


OBSERVATIONS ON POLYCYSTID GREGARINES 
FROM ARTHROPODA * 


MINNIE E. WatTSOoN 


The following pages contain observations on already known and 
new species obtained by the writer chiefly during the past summer. 
BULBOCEPHALUS WARDI NOV. geil., NOV. Spec. 
[Figures 1, 2, and 3] 
Host: Clerid larva (Det. E. P. Felt) 
Habitat: Intestine 
Location: Oyster Bay, Long Island, June, 1916 

Fifty or more specimens of this gregarine, mostly free trophozoites, 
were taken from a single Clerid larva. The sporonts are solitary and 
rather small, and the species is characterized chiefly by the distinct 
bulb in the mid-region of the large epimerite of the trophozoite. 

The longest sporont seen measured 190 in length and 45y in width. 
One trophozoite exceeded in length the longest sporont, being 290, in 
length. The protomerite of the sporont is slightly longer than wide, 
broadly rounded in front, and slightly constricted at the septum. The 
deutomerite is widest at the “shoulder” and tapers from thence, ending 
in a long blunt-pointed extremity. The average ratio of length of 
protomerite to total length is about 1:5; the protomerite and deuto-. 
merite are of approximately the same width. 

The epimerite is unique. It consists of three parts: a, a broad base 
fitting on the apex of the protomerite, tapering at the top to form a 
neck; b, a spherical bulb, in the middle; c, a stout style at the apex. 
In other words, the epimerite consists of a stout broad-based style, 
bulbous in the middle. In length, it measures one-fifth to one-third the 
total length of the trophozoite. Complete dimensions of several epi- 
merites are given in the table at the end of this species. 

The protoplasm is dense, rendering it dark in color in both proto- 
merite and deutomerite. The ellipsoidal nucleus is not visible in the 
live sporont ; when stained, it is seen to be small in the sporont (larger 
in comparison in the trophozoite), and lies diagonally across the largest 
part of the deutomerite. One karyosome is present. ne 

The writer is unable to classify this species in any known genus. 
The epimerite is most nearly related to that of the genus Stylocephalus ; 
in this genus it consists of a more or less long, slender neck terminating 


* Contributions from the Zoological Laboratory of the University of Illinois 
under the Direction of Henry B. Ward, No. 79. 


66 THE SOURNAL OF PARASITOLOGY 


in a large or small papilla. There may or may not be a bulbous base. 
The nucleus is ellipsoidal. The type species of the genus is Stylo- 
cephalus oblongatus (Hammerschmidt) Watson, see Watson 1916: 159. 

The present specimens differ from the type species of the aforesaid 
genus in but one respect: The bulbous papilla of the epimerite is 
placed not at the apex of the cylindrical neck, but at its mid-point. In 
no species of the named genus is there a papilla at other than the apex. 
It therefore devolves upon the writer to describe a new genus in order 
to place the specimens found. | therefore designate it Bulbocephalus, 
and place it as follows: 


Family Stylocephalidae Ellis. Sporonts solitary, epimerites varied. Nucleus 
ovoidal. Dehiscence Iby pseudocyst. Spores irregularly shaped, brown or 
black, in chains (Watson 1916: 47). 

Type Genus Stylocephalus Ellis. Epimerite a dilated papilla at the 
end of a long slender neck. Cyst covered with small papillae and 
indentations. Spores hat-shaped. 

New Genus Bulbocephalus. Epimerite a dilated papilla placed in the 
middle of a rather long slender neck. Cyst and spores unknown. 


The species is named for Professor Henry B. Ward of the Univer- 
sity of Illinois. 
A table of typical measurements, in microns, is given herewith: 


Ene timepimlenttesneiayeeits «ses ses + ae 60 x x 40 30 
Length protomerite (without epimerite) . | 50 3 40 15 20 
Menethwdeutomentlente sia... /v os,» «7 ean 180 62 30 So 40 
TRoralMlenothmspeLoiteees +s -1-- 6-6 «10 6 +r 290 192 170 110 90 
Waidthiiepiinerite Wepinec eh. sc scsleee es + os 16 x x 16 10 
Widtheprotomentiemmyeee: =~ o24 0c. se. ee 42 32 40 20 18 
Wadtit deutomierite Aes. ... .<as'e er. s,s 37 45 40 28 31 
Ratio length prot. (without epim.) : 

forall EMERG. at Peehe +c void Boe ge ke os 58 deoAen de ay yee 1:45 
Ration widtheprotcmwidin dettt....- 4... ee Ss laird on sales 11:4. Veale 
DidineteGenlUGLENSMAEPI occ chee secs sce Nlesaem Sxl 5 eae. 11x27 


BULBOCEPHALUS ELONGATUS nov. spec. 
[Figures 4, 5, and 6] 
Host: Cucujus larva (Det. Adam Boving) 
Habitat: Intestine 
Location: Oyster Bay, Long Island, August, 1916 


Specimens of this peculiar gregarine were very abundant in the one 
beetle larva obtained. The sporonts are relatively very long and 
slender, unlike those of any gregarine before taken by the writer. The 
maximum length recorded for a sporont is 6004; the maximum width, 
50u. The ratio of length of protomerite to total length is 1:11, and 
the ratio of width of protomerite to width of deutomerite about 1: 1. 

The protomerite is widest near the septum, at which there is a slight 
constriction, and the deutomerite is widest at the “shoulder,” gradually 
tapering from thence to form a very long posterior extremity. The 
protoplasm is dense in the sporont, but much less so at the posterior 


WATSON—POLYCYSTID GREGARINES 67 


end, and the nucleus is obscured in the adult. In the trophozoite, it is 
seen to be small and ellipsoidal and placed diagonally to the long axis 
of the animal. 

The epimerite is a rather long, stout style dilated in the middle to 
form a good sized papilla. The shape of the epimerite indicates that 
this species is closely allied to the preceding, and is therefore placed 
in the new genus, Bulbocephalus. 

A table of a few measurements in microns follows: 


PIETER OMOPUNC. «Wp osycbise dea «56a ohn oe a2 x x 40 
Length protomerite (without epimerite). .. 47 30 20 
emeOMETGOMCIILG .cc.2 20k es cass cee enaee 553 300 170 
Motlenoth: Sporonteswks «yop ots has oo Scorers 600 330 230 
WV TERME DIIMELILED sec se Sess o'er ia sale eleieye x a 10 
Mi edit. prOLeitierite®. obo. ee a0.5 se. eg ows 35 40 20 
WWE NGeNtOmenites srlccpvee co <fo9 sider anit 30 50 23 
Ratio length prot.: total length............ NS ino hese 
RavoOnwidth prats., width dent. <.2<- 2.2m esd 1 Ps I ett 


DaIMeTeE AILIGLIEIIG Nae bie oe acs arsine recto ten dl a ieee 


PYXINIA BULBIFERA 10V. spec. 
[Figures 7, 8, 9, 10, 11 and 12] 
Host: Dermestes lardarius Linn. (Det. E. P. Felt) 
Region of infection: Mid-intestine 
Location: Oyster Bay, Long Island, May, 1916 


Fifty or more specimens of this parasite were taken from one adult 
larder beetle, many of them being free trophozoites. 

The sporonts are solitary and long and slender in shape, except at 
the “shoulder,” where they are appreciably wider (Figs. 7 and 8). The 
largest sporont seen measured 850u long and 160u wide; the smallest 
specimen without an epimerite, 600u long. The average ratio of length 
of protomerite to total length of sporont is 1:5, and the ratic of width 
of protomerite to width of deutomerite is 1: 1.3. 

The protomerite is generally of the same width as height, infre- 
quently a little wider than high. It is bluntly cone-shaped, the widest 
portion lying two-thirds the distance from the apex, being constricted 
below this to meet the septum. There is often a slight indentation at 
the apex, left by the dropping off of the epimerite. The epicyte forms 
a very thick layer over the end of the protomerite (Fig. 9). 

The epimerite consists of two parts, a stout bulbous crenulate and 
crateriform base and a short thick style. The bulbous base was fre- 
quently seen to expand and contract with considerable force by means 
of longitudinal folds which open and collapse, the motion of the 
myonemes probably forcing protoplasm in and out of the epimerite 
from the protomerite (Figs. 10, 11 and 12). The epimerite measures 
from 60y to 100u in length on large free trophozoites. The largest epi- 
merite seen measured as follows: Style, 70u long and 10p wide at its 
base; bulb 30u high and 35 wide at its widest part. 


68 THE JOURNAL OF PARASITOLOGY 


The deutomerite tapers gradually from the “shoulder,” ending in a 
blunt point. 

The protoplasm is dense in most parts of the body. The widest 
portion of the deutomerite is very dark brown, nearly black, the pos- 
terior end being lighter in color. The protoplasm gradually becomes 
less dense in the latter region and the granules cease entirely some 
distance before the end is reached. The protomerite is tan in color and 
the epimerite transparent. 

The nucleus is scarcely visible in an unstaiued sporont, but in 
stained specimens is seen to be an ellipsoidal body lying always at right 
angles to the main axis of the body and never diagonally, as is often 
the case in gregarines: A single large karyosome is present. The 
nucleus measured in a trophozoite about 90 by 40u. 

Myonemes were visible at the end of the deutomerite and in the 
protomerite with a magnification of 490; longitudinal striations were 
seen with a magnification of 770 and intense transmitted light. 
Brownian movement was noted where the protoplasm was least dense, 
viz., at the apex of the protomerite and the tip of the deutomerite. 

Neither cysts nor spores were seen. 

Although species of the genus Pyxinia have been described from 
the genus Dermestes, the present specimens do not fit any of these 
descriptions. They differ from Pyxinia rubecula Hammerschmidt 
(Léger, 1892: 140) chiefly in shape of the epimerite, which in the latter 
species is urn-shaped with a wide mouth and a crenulate periphery, 
and with a short style. Léger’s figure for an epimerite of this species 
indicates the diameter of the urn to be five times its depth and the 
minute style so short that it scarcely projects beyond the rim. The 
present specimens indicate an urn narrower at the periphery than else- 
where and a style nearly as wide at its base as the encircling crenulate 
rim. In shape and proportions, the two species compare favorably, 
and the host is a beetle of the same genus, although previously 
described from Europe. 

The conspicuous refractile pyxinin crystals of P. crystalligera 
Frenzel (1892: 314) are lacking in the present species, and the long, 
slender sporonts with bulbous protomerites contrast strongly with the 
species described above. Maximum length of sporonts and shape of 
the epimerite are similar in the two. 

The other two described species of Pyxinia, P. frenzeli Laveran 
and Mesnil, and P. mébuszi Léger and Duboscq (Watson, 1916: 151) 
are radically different from this species in size and shape of the 
sporonts and in the character of the epimerite. 


_ 


WATSON—POLYCYSTID GREGARINES 69 


Dimensions of a number of typical specimens are appended here- 
with, measurements being in microns: 


TctiGtly CORMELILE,.. dca as ee qos oneal nees x = 40 x 100 
Length protomerite (without epimerite). 150 150 150 160 140 
Menath- devitomerite .4...09 4. + case seein. 700 690 600 580 500 
Pe NCR Ta Por. bcs’. as aie Smee eee een 850 840 750 740 640 
With protomerite ....ovcs leas x seen vs 150 150 140 110 120 
Wat HewgmOnienite .. 16.66 cs cess wena 160 200 190 150 150 
Ratio length prot. (without epim.): 

LOLA Venotlt ves «0 .sdace hi pnet ele ilculy fon" eeu soya ian Midies 14160 21416 
Ratio width prot.: width deut.......... Peal elese esis edges ea 


ree Ler TIMIGIETIS! |. «sca Ss,c. cette eee inte stedar ar 


GREGARINA NEGLECTA 10V. Spec. 
[Figures 13, 14, 15, and 16] 


Host: Ceuthophilus neglectus Scudder (Det. A. N. Caudell) 
Habitat: Intestine 
Location: Oyster Bay, Long Island, August, 1916 


Four associations of this gregarine were found in the mid-intestine 
of one specimen of this camel cricket. Although very similar to many 
other species of the genus Gregarina in shape, this species may be 
characterized by a papillate protomerite in the primite and a perfectly 
egg-shaped satellite, there being no constriction at the septum. 

The associations seen were all of about the same length, the maxi- 
mum length of an association being 9004; that for a single -sporont 
500n. The maximum width measured was 230y. In every instance, 
the primite was longer than the satellite, maximum length of the satel- 
lite being 430u. The average ratio of length of protomerite to total 
length of the primite is 1:6; for the satellite the average is about 1:8. 

The primite is elongate-ellipsoidal in shape and the satellite ovate, 
widest anteriad. The protomerite of the primite is broadly rounded, 
widest at the base and papillate at the apex; it is approximately one 
and two-thirds times as wide as high. There is a slight constriction at 
the septum. The deutomerite is regularly ellipsoidal, terminating in a 
broadly rounded extremity. 

The protomerite of the satellite is very broadly rounded anteriorly, 
but with a trace of the papilla so prominent in the primite. It is three 
times as wide as high and is widest at the septum; there is no constric- 
tion at the septum. The deutomerite is widest near the middle and 
terminates in an extremity less well rounded than in the primite. 

This species is very dark brown in color with the protomerite 
slightly less dense. In the satellite, both protomerite and deutomerite 
are very dark, rendering the septum difficult to trace. The nucleus is 
completely obliterated in the living animal. The epicyte is very thin, 
much less so than is usual with this genus. 


EXPLANATION OF PLATE 


Fig. 1—Sporont of Bulbocephalus wardt. 

Fig. 2—Large free trophozoite of B. wardi, with epimerite. 
Fig. 3—Smaller trophozoite of B. wardi. 

Figs. 4, 5—Sporonts of Bulbocephalus elongatus. 

Fig. 6—Free trophozoite of B. elongatus. 

Figs. 7, 8—Sporonts of Py-vinia bulbifera. 


Fig. 9—Protomerite of P. bulbifera, to indicate thickening of epicyte and 
indentation left by epimerite. 


Fig. 10.—Epimerite of P. bulbifera, not expanded. 

Fig. 11—Epimerite of P. bulbifera, with bulb partially expanded. 
Fig. 12—Epimerite of P. bulbifera, bulb fully expanded. 

Figs. 13, 14——Two associations of Gregarina neglecta. 


Fig. 15——Protomerite of primite of G. neglecta, to indicate papillate apex 
and thickening of epicyte. 


Fig. 16—Protomerite of satellite of G. neglecta. 

Figs. 17, 18—Two associations of Gregarina polymorpha. 
Fig. 19—Protomerite of primite of G. polymorpha. 

Fig. 20.—Protomerite of satellite of G. polymorpha. 
Fig. 21—Association of Gregarina blattarum. 


Fig, 22.—Protomerite of primite of G. blattarum, to show thickening of 
epicyte at apex. 


Fig. 23—Protomerite of satellite of G. blattarum. 


SAAT Li 


O48 


oo 


WATSON—POLYCYSTID GREGARINES 71 


Cysts were found in three stages of development, measuring in 
internal diameter about 300u (see table of dimensions). No spores 
were seen. 

Gregarines have been described from the genus Ceuthophilus in 
three cases: two by Ellis, and one by myself (Watson, 1916: 114-5). 
The present species is differentiated from the first two species in shape, 
from the one in shapes of the two protomerites in the association, from 
the other in shapes of the deutomerites. It differs from the last 
described species in size of sporonts and cysts and in relative thickness 
of the epicyte, the associations of the present species being approxi- 
mately three times the length of those of the species formerly described, 
and the cysts twice as large in the present species (150u and 300, in 
diameter, respectively ). 

A table of measurements is appended, dimensions being stated in 
microns : 


Primite Satellite 
OF | <a TS 
a b Cc d a b c d 
Length protomerite .... 70 100 70 70 50 40 50 60 
Length deutomerite .... 430 400 390 400 350 330 370 370 
Width protomerite ..... 130 150 120 120 170 190 130 150 
Width deutomerite ..... 210 230 210 190 200 210 200 200 


Total length sporont.... 500 500 460 470 380 370 420 430 
Ratio length prot.: total 


ene than terse at acta Vet bey ena Brae feb 12921784 Le fA 
Ratio width prot.: width 
MEE tse tee eTocs Ue dee eed 5) Mon ect a 12; Led 31:15), leh3s 


Total length association. 880 870 880 900 
Diameter cysts: 

Without envelope... 300 310 290 

With envelope...... none 470 none 


GREGARINA POLYMORPHIA (Hammerschmidt) Stein 
[Figures 17, 18, 19, and 20] 
Host: Larva of a Tenebrionid, probably 7. molitor 
Region of infection: Intestine 
Location: Oyster Bay, Long Island, August, 1916 


One specimen of this Tenebrionid beetle larva from a meal sack 
was found to contain half a dozen associations of one of the gregarines. 

The specimens are characterized by a regularly curved protomerite 
in the primite and a protomerite of the same shape, but slightly flat- 
tened, in the satellite. 

The sporonts vary in length from 280p to 320u, in width from 
1002 to 120u. The ratio of length of protomerite to total length of 
sporont is approximately 1:7 for both primite and satellite; the ratio 
for width of protomerite to width of deutomerite is about 1: 2. 

The sporonts are small, elongate ellipsoidal with a relatively small 
protomerite. The protomerite is beautifully rounded in outline in the 
primite, representing about two-thirds of a sphere. The widest part 


72 THE JOURNAL OF PARASITOLOGY 


is just below the middle, and there is a distinct constriction at the 
septum. The protomerite of the primite is approximately as wide as 
high; while that of the satellite, which is the same shape but flattened 
at the apex, is wider than high. The deutomerite is widest at about its 
mid-region, tapering anteriorly to the septum and posteriorly, ending 
in a long blunt cone. 

The protoplasm in the sporont is abundant, rendering it dark brown, 
almost black in the deutomerite and dark tan in the protomerite. No 
nucleus is visible in either member of the adult association. The ecto- 
sarc is especially thin, even at the tip of the protomerite and at the 
septum, causing rupture to be easily effected in water, an unnatural 
medium. 

The animals are relatively active in movement, both of progression 
and contortion. : 

Neither cysts nor spores were seen. 

This species has been described from Europe and Japan, but not 
heretofore from the United States. The first mention of the species 
was made in 1838 by Hammerschmidt, and it has been seen at various 
times since then, by Stein, Frantzius, Schneider, Léger and Duboscq, 
Ishii, etc.; see Watson, 1916:172, for correlations and references. 
No two descriptions and drawings tally exactly, but there is not suff- 
cient evidence to differentiate the observations into separate species. 
The present detailed description for a known species is given because 
here also are indicated variations from the orginal of Hammerschmidt, 
and since the specimens seen by every worker differ slightly from those 
of the others, it does not seem wise even to describe a new variety. 

A table of a few typical measurements follows, dimensions being 
in microns: 


Primite Satellite 
ae SS SS ee 
a b c d a b c d 
Length protomerite ..... 60 40 40 40 40 40 40 40 
Length deutomerite .... 250 270 240 280 280 280 240 270 
Width protomerite ..... 60 50 50 40 70 60 40 60 
Width deutomerite ..... 110 110 100 120 120 110 100 120 


Total length sporont.... 310 310 280 320 320 320 280 310 
Total length association. 630 630 560 630 
Ratio length prot.: total 


length! .282 anes « 1:51. 1:7.7 1:7 1:8 1:8>04¢8 25 eee 
Ratio width prot.: width P 
deat taser ee ss 1:18 1:22 1:2 41:3-° 1:17 Tt: 


GREGARINA BARBARARA Watson 1915 
This species was found at Oyster Bay, Long Island, in Adalia 
bipunctata in June, 1916. This, our commonest lady beetle in the East, 
was examined frequently throughout the year, but was found para- 
sitized but once. Two associations and half a dozen isolated sporonts 


. 


WATSON—POLYCYSTID GREGARINES 73 


were found. This species has heretofore been recorded from an 
unidentified Coccinella, not the present species. 

A few measurements, in microns, are appended to supplement those 
already given (Watson, 1915:31 and 1916:185), and for sporonts 
somewhat smaller than those previously described. 


Primite Satellite 

i 

a b c d a 
Meneth Protomerite: ..4. cect cas See oh 20 30 20 20 8 
Weneth CeglOmerite ys.9< sien cs sak oe ae 80 70 70 80 52 
With, pratemnevrite. «. tind: vichsle os tee ; 20 30 30 30 30 
MAME CeHbOMmerthes.. Ach ies sols satin a bye 55 60 45 55 40 
otal length sporontsn...t =. 202s. oe te 100 100 90 90 60 
fetal) letieth *associatione.ts..ic0e.tale ee 160 
Ratio length prot.: total length........... Sterol 45° 125 i aay as 
mauG With pret widen cette. cee, oe P20 12 15. 921.695 1:13 
Piameter- nucleutse.\ntrs ees ste os sede ots Pelz 


GREGARINA BLATTARUM Siebold 
[Figures 21, 22, and 23] 


Host: Blatta ortentalis Linn 
Regions of infection: Intestine and rectum 
Location: Urbana, Illinois, June, 1915 


A dozen or more biassociative gregarines were found in one speci- 
men of the Oriental cockroach, this being the greatest number found 
by the writer in a single host. Many roaches were examined with 
negative results. 

The insects are also parasitized by two species of nematodes, an 
infusorian, and an amoeba, the last two of which were described by 
Leidy in 1877 and 1881. 

This gregarine is characterized by long, slender sporonts, blunt at 
the posterior end, by a conical, or papillated, protomerite in the primite, 
and a broad, flattened protomerite in the satellite. 

The sporonts vary in length from 510 to 1100p, and in width from 
160u to 400u. The average ratio of length of protomerite to total 
length of sporont is 1: 5 for the primite and 1:8 for the satellite. The 
protomerite of the primite is approximately two-thirds as wide as the 
deutomerite, that of the satellite approximately three-fourths to fully 
as wide as the deutomerite. The ratio in the primite of width of pro- 
tomerite to width of deutomerite is about 1: 1.7, in the satellite about 
hl a : 

A table of representative measurements is appended herewith. 

The protomerite of the primite is bluntly pointed, the ectoplasm at 
the apex being a much thicker layer than elsewhere in the animal. The 
widest portion of the protomerite is about two-thirds of its length from 
the apex, and there is a slight constriction at the septum separating 
protomerite and deutomerite. 


74 THE JOURNAL OF PARASITOLOGY 


The deutomerite is elongate ellipsoidal, varying but little in width 
throughout the length, and broadly rounded at its posterior end. The 
end attached to the satellite is but little flattened. 

The protomerite of the satellite is slightly flattened anteriorly, and 
there is but little or no constriction at the septum. The deutomerite is 
more or less irregularly shaped, ending in a rather blunt point. 

The nucleus is small and spherical. It measures about 90 in 
diameter in sporonts. The spherical karyosome is faintly visible. 

The deutomerite of the sporonts is very dense and blackish, the 
protomerite slightly less dense and dark tan in color. The deutomerite 
is finely granular and homogeneous, and is slightly more dense in the 
satellite than in the primite. The nucleus was not visible in the satellite 
of any specimen seen. The protomerite contains large spherical masses 
less closely packed together than in the deutomerite. 

This species was found first by Siebold in Germany, and by many 
subsequent workers, including Frantzius, Leidy, Schneider, Marshall, 
de Magalhaes, and Crawley, and from Germany, France, Brazil, and 
Pennsylvania (see Watson, 1916: 99-100). 

The present specimens have the same general proportions as those 
already described, but reach a much greater length than that stated 
by Leidy, which is 500, for a single sporont; no other writer has given 
dimensions. The specimens now described are undoubtedly closely 
related to the species he saw and described as Gregarina Dlattae. 
orientalis, for he mentions the “slight papillary thickening of the . 
integument” at the apex of the protomerite and indicates this feature 
in his three drawings. He does not state, however, whether or not the 
species is associative. 

Because of the very considerable confusion surrounding this 
classical species in the past, it does not seem to the writer wise to add 
to it and describe the specimens now found as a variety of the type 
species when the only disparity is found in a papillate or non-papillate 
apex of the protomerite. A number of variations have already been 
described, but are now separated into species. 

The measurements given below are in microns. 


Primite Satellite 
———— a Ua ae 
a b c d a b c d 
Length protomerite ..... 120 150 160 200 60 80 100 160 
Length deutomerite .... 390 720 790 900 460 750 600 870 
Width protomerite ..... 120 160 200 200 150 160 250 250 
Width deutomerite ..... 200 260 300 360 200 230 250 400 


Total length sporont.... 510 870 950 1100 520 830 800. 1030 
Ratio length prot.: total 


lengtht se" jonueec as 1:42 1:58 1:59 1:55 1:8651310 “1-3 ee 
Ratio width prot.: width 
dents cma peeeneses 1:1,7. 1216) 4:15). 2:18. eel ae 


Diameter nucleus ...... .... 90 
Total length association. 1030 1700 1750 2130 


aa 


WATSON—POLYCYSTID GREGARINES 75 


A new place record has been established for Leidyana erratica 
(Crawley) Watson, at Douglas Lake, Michigan, by Mr. H. G. May, 
of the University of Illinois. This species and Gregarina oviceps 
Diesing were very commonly parasitic in the same host, which has been 
designated Gryllus americanus Blatch. Mr. May says, however, that 
the host may be G. pennsylvanicus Burm., or even an hitherto unde- 
scribed species of cricket. The cricket fits neither description perfectly. 


In none of the instances recorded above was it possible to complete 
a life-history of the parasite because the hosts are for the most part 
uncommon and rarely more than one specimen of the same species was 
taken during the summer. It was unfortunately impossible to secure 
any intestines for sectioning purposes and, as cysts were rarely seen in 
the host parasitized, the life-histories were not carried to completion. 

Type specimens of the above species have been deposited in the 
Ward Collection of Parasites at the University of Illinois. 


SUMMARY 


A new genus, Bulbocephalus, with two new species is described for 
the family Stylocephalidae. 

New species are described for Pyxinia and Gregarina, and new data 
are given for Gregarines already known. 


REFERENCES CITED 


Frenzel, J. 1892. Ueber einige argentinische Gregarinen. Jen. Zeitschr., 
27 : 233-336; 1 pl. 

Léger, Louis. 1892. Recherches sur les grégarines. Tabl. zool., 3: 1-183; 
2A aplk 

Watson, M. E. 1915. Some New Gregarine Parasites from Arthropoda. 
Jour,’ Parasit., 2: 27-30; 2 pl. 

1916. Studies on Gregarines. Ill. Biol. Monographs, Vol. 2, No. 3, 258 pp.; 
15 pl. 


ON A TREMATODE LARVA ENCYSTED IN A CRAB; 
HELICE TRIDENS (DE TAA 


SapAao YOSHIDA ‘ 
Pathological Department of Osaka Medical Academy 


In November, 1915, a considerable number of the encysted larvae 
of a trematode and some specimens of an infected crab were for- 
warded for identification by T. Urita, a friend of mine in Kagoshima, 
with a letter which read as follows: “The liver and the inner surface 
of carapace of a crab, Helice tridens (de Haan), are heavily infested 
with the egg-like cysts with thick, transparent wall containing a moving 
worm which seems to be a trematode larva. Some worms creep out. 
The crabs are found abundantly in the salt-field living in the holes they 
dig. Seventy to ninety per cent or more among the crabs in the vicinity 
of Kagoshima City are infected with the cysts.” 

Afterwards I obtained the crabs of Kagoshima Prefecture on 
several occasions from the same friend and from H. Yamakado, a 
student of our academy, and collected them myself on the seashore 
near Osaka. The crabs are found in various districts, especially in 
the warm parts of this country. They live in ground near the seashore, 
such as salt-fields, river mouths, and other coast areas exposed when 
the tide ebbs, and habitually build burrows in which they live. The 
crabs are not used as food in Japan proper, but they are said to be 
eaten in some parts of Korea and Formosa. ‘here is no need to 
describe the morphological characters of the crab here. 

On several occasions I examined two hundred and fifty specimens 
of the crabs and. found about 90 per cent of them infected with the 
cysts. There is no local difference in frequency of infection between 
the crabs collected near Osaka and those of Kagoshima. The number 
of cysts harbored in a crab is variable but generally large, varying 
from one to several hundreds, though sometimes they are in masses by 
the thousand. The encysted larvae occur generally in the ovary, on 
the wall of the stomach, in the hypodermis lining the carapace, and 
other parts of the body cavity of the host. The ovary (Fig. 14) and 
hypodermis are most heavily and frequently infested with them. In 
the liver I have rarely found the cysts, although T. Urita informed 
me of the liver infection in his letter quoted above. I found occa- 
sionally, however, infected pieces of membranous tissues twining round 
the lobes of the liver, a condition which perhaps would suggest liver 
infection. 


YOSHIDA—TREMATODE LARVA ENCYSTED IN CRAB 77 


Morphology of the Encysted Larvae —tThe encysted larvae found 
in the crab are oval or rarely spherical in shape, varying more or less 
in size. Measurements of eight cysts show a range in length from 
0.407 mm. to 0.601 mm., and in breadth from 0.366 mm. to 0.510 mm. 
The average length was 0.521 mm. and breadth, 0.418 mm. 

A slight pressure by the cover-glass may alter the dimensions of 
the cysts; the measurements given above are obtained from those not 
compressed. 

The wall of the cyst is a transparent chitincus membrane from 
0.02 to 0.04 mm. thick through which the larva may be readily 
observed. An actively moving larva in the fully developed cyst is light 
brown in color with dark spots marking the position of the yolk glands. 
The worm occupies almost all the space within the cyst with the body 
bent in various fashion (Figs. 1 B, C). In the most common position 
in the cyst, either the anterior or posterior extremities of the body, or 
both, coil ventrad, and both lateral margins of the anterior region are 
also sometimes bent over the ventral surface. Through the wall of the 
cyst one may recognize the organs of the larva, i. e., oral sucker, ali- 
mentary tract including pharynx, long esophagus, intestinal ceca, 
excretory vesicle, and such genital organs as ovary, yolk glands and 
testes; but the identification of these organs is complicated by the 
movements of the body. 

The encysted larvae in full development may emerge from the 
cysts at various times after the latter are removed from the crab and 
put in distilled water or physiological salt solution. Sometimes this 
takes place within only ten to thirty minutes after the cysts are 
removed from the host. Cysts from dead crabs are generally some- 
what weakened, in consequence of which the larva may easily emerge 
from the cyst. Encysted larvae from crabs that have been dead a long 
time are also dead or so strongly affected as to be at the point of 
death. It is believed that the action of the putrescent fluids explains 
the fact that the larvae are generally found out of the cyst and dead. 
During my study I often found the larvae free and dead after a day 
or two in culture dishes. 

External Feature of the Larva Liberated from the Cyst—The larva 
freed from a fresh cyst is actively mobile, changing its shape and size 
greatly, especially in the anterior region. Figure 2 B serves to show 
the extent of change in the outline of a worm in the contracted state. 
On the whole, it is concave on the ventral surface and convex on the 
dorsal; an accurate measurement of length and breadth is very diffi- 
cult when the worm is alive. The most natural form, however, is to 
be found in a larva that has recently died a natural death. The larva 
put between the slide and cover-glass also assumes a form resembling 


78 THE JOURNAL OF PARASITOLOGY 


the natural, though slightly exaggerated. The most common form then 
assumed is an elongated oval, the middle one-half or two thirds of the 
body length being of nearly uniform breadth. Both extremities taper, 
the posterior being more obtuse than the anterior end. In seven fixed 
specimens measured, the length varied from 0.533 to 0.833 mm., and 
the breadth from 0.283 to 0.325 mm., the average dimensions being 
0.701 by 0.301 mm. Two living specimens measured, respectively, 
1.0 by 0.50 mm. and 0.922 by 0.43 mm. Four specimens compressed 
and mounted in potassium acetate varied from 0.9 by 0.466 mm. to 
1.18 by 0.56 mm., with an average of 1.083 by 0.515 mm. Five speci- 
mens compressed and mounted in Canada balsam varied from 0.75 by 
0.35 mm. to 1.07 by 0.50 mm., with an average of 0.918 by 0.438 mm. 


Fig. 1—A, portion of the crab’s ovary infested with cysts. X23. B, ce 
two encysted larvae. > 60. 


The entire surface of the body is armed with minute spines which 
are denser in the anterior region than in the posterior and inconspicu- 
ous in mounted specimens. The oral sucker (Fig. 2 B) is subterminal 
and spherical or ovoid in shape; its aperture varies according to the 
state of contraction of the organ. The ventral sucker (8) situated 
about one-third the body length from the caudal end is oval or spherical 
and smaller than the oral sucker. In the living specimen it is usually 
indistinct. “The average dimensions of the oral sucker in compressed 
living specimens were 0.053 by 0.05 mm.; in specimens mounted in 
potassium acetate, 0.0693 by 0.0625 mm.; in specimens mounted in 
Canada balsam, 0.052 by 0.055 mm. The average dimensions of the 
ventral sucker in compressed living specimens were 0.037 by 0.024 mm. ;” 
in specimens mounted in potassium acetate, 0.035 by 0.040 mm. 

Internal Structure —The internal structure of the larva (Fig. 2) is 
easily observed in the living specimens compressed or in the mounted 


YOSHIDA—TREMATODE LARVA ENCYSTED IN CRAB 79 


specimens. The larva is femarkable in that almost all the essential 
genital organs are well developed. 

The prepharynx (P) is distinct but short and slender, 0.03 to 
0.08 mm. long and 0.01 to 0.02 mm. broad. The pharynx (/7) is sub- 
spherical, 0.035 to 0.05 mm. long and 0.025 to 0.048 mm. broad. The 
esophagus (£) is very long and slender, 0.25 to 0.35 mm. long, with 
the breadth gradually increasing posteriorly to a maximum of 0.015 
to 0.027 mm. The intestinal ceca (C) are short, running from the 
posterior end of the esophagus obliquely postero-lateral, so as to form 
a V-shape, the distal ends being far from the lateral margins of the 
body. The length of the ceca on both sides is nearly equal in the same 
individual and slightly variable in the different specimens (0.2 to 


igs): R C 


Fig. 2—A, specimen mounted in potassium acetate. 60. 8B, living larva 
in motion. 60. C, semilunar organ and ventral sucker. > 280. 


0.3 mm. long). The breadth is usually maximum at the middle part 
of the cecum, but it may be variable according to the condition of its 
contents. 

The excretory vesicle (V) is V- or Y-shaped, and each inner end 
gives off two branches. The winding and branching canals arise from 
the anterior end of the vesicle, but their entire course cannot be made 
out distinctly. ; 

The cerebral ganglion (N) surrounding the prepharynx and the 
lateral nerves from it may be observed in good specimens. 

Genital Organs.—Two testes (T) lie side by side near the posterior 
end of the body, just in front of the excretory vesicle. They are oval 
and subequal with the long axes transverse or obliquely to the axis. 


80 THE JOURNAL OF: PARASITOLOGY 


They measure 0.15 to 0.26 mm. in length and 0.09 to 0.16 mm. in 
breadth. The vas deferens arises from the mesial surface of each 
testis, but its further course cannot be traced. Just anterior to the 
ventral sucker there is a peculiar organ (B) resembling the cirrus sac. 
It is semilunar, lying transversely with concave side posterior; each 
extremity is provided with a chitinous process of form unlike the 
other. The chitinous process on one side (usually the right) tapers 
from its wide base and terminates in a smaller single tubular end, 
while the distal end of the process on the other side is divided into 
smaller unequal shaped processes (Fig. 2 C). The free ends of both 
chitinous processes face each other across the ventral sucker. The 
significance of this semilunar organ cannot be determined and no con- 
nection with other organs was determined. Before and behind the 
ventral sucker there are muscle fibers connecting with the semilunar 
organ. 

The ovary (QO) lies on one side (usually the right) of the median 
line and in the space bounded by the semilunar organ, the posterior 
end of the intestinal cecum, and the testis on the same side. It is oval 
or spherical in shape, being 0.07 to 0.12 mm. long and 0.07 to 0.1 mm. 
broad. The short oviduct springs from its postero-mesial aspect and 
runs postero-mediad to a cell-group situated in the space between both 
testes and the semilunar organ. This probably represents the shell 
gland and the uterus. Here one may find granules very similar when 
stained to the yolk granules. The cell-group is deeply stained by 
carmine. 

The yolk glands (¥Y) occupy a small area on the antero-lateral 
aspect of each cecum. The glands on each side consist of from nine 
to eleven follicles on the ovarian side and from seven to nine on the 
other side. These follicles are irregular in form and vary slightly in 
size; but the maximum diameter varies between 0.055 and 0.08 mm. 
and the minimum between 0.027 and 0.045 mm. The yolk duct (D) 
from the glands runs postero-mediad to the cell-group mentioned 
above. The genital aperture could not be detected. 

From the characteristics mentioned above it is difficult to determine 
the adult form corresponding to the encysted larvae. I endeavored to 
find the matured form by animal experiments. On November 26, 1915, 
many cysts from the crabs were fed to three guinea-pigs, and on 
December 14 a number of others to two young cats. These animals 
died or were killed after several days and were carefully examined 
for distomes, but in vain. On February 5, 1916, several cysts were 
eaten by three guinea-pigs, but they did not develop. On April 25 cysts 
were given to five frogs. After four or five days the frogs were killed 
and examined for the distomes. In the stomach, intestine, and cloaca 


YOSHIDA—TREMATODE LARVA ENCYSTED IN CRAB 8] 


larvae were found free from the cysts and dead. Thus far in animal 
experiments I have not succeeded in obtaining the adult. 

These encysted larvae evidently differ from the adult less than in 
most other cases, because the genital organs are so well developed. 
The alimentary tract, long esophagus, short intestinal ceca and the 
position of the yolk glands suggest a relationship to the genus Brachy- 
coelium. Other feeding experiments with the encysted larvae are 
under way at present and will be described later tully with the micro- 
scopical structure of the worm. 


CYTOLEICHUS PENROSEI, A NEW-ARACHNOID PARA- 
SITEePOUNDEIN, THE DISEASED ;LUNGSZOR 
A PRATRIE «DOG, CYNOMYS 
LUDOVICIANUS 


Frep D. WEIDMAN 


On March 28, 1907, a male prairie dog, Cynomys ludovicianus, 
died in the Philadelphia Zoological Gardens with acute broncho- 
pneumonia. The inflammatory condition was recognized at the time 
of autopsy, the lungs being described as diffusely red, standing out 
well, their apical parts emphysematous and cut surface showing minute 
white projecting areas. The parasites forming the basis of this com- 
munication were not recognized at this time om account of their 
extremely small size, being among the smallest of which the writer has 
found record, and much smaller than the ones which were found here 
(Weidman, 1915) in the lungs of a monkey and upon birds. The skin 
bore, especially around the head and shoulders, brownish crusts which 
were tightly adherent to deeper parts and which, when examined micro- 
scopically, were found to contain a fungus. 

The gross diagnosis. of bronchopneumonia was confirmed micro- 
scopically, shown by the presence of numerous red blood cells, fre- 
quent clumps of fibrin and moderate numbers of leukocytes in the air 
sacs, with which a greenish brown granular material, doubtless the 
excrement of the mites, was intermixed. In addition to the inflamma- 
tory disease, the sections showed a high grade of emphysema and 
bronchiectasis (Figs. 14, 15 and 16). 

That articulated parasites were also present was recognized in the 
microscopical sections which shortly and routinely followed the autopsy 
in 1907, but it was only during a recent review of prairie dog tissues 
in connection with another parasite of prairie dogs | Hepaticola hepatica 
(Bancroft, 1893) Hall, 1916] that their arachnoid nature was deter- 
mined. During this interval (about nine years) the tissue had been 
preserved in alcohol, precluding experiments on transmission of the 
infestation and observation of living specimens, but not interfering 
with staining qualities of sections. These, in favorable cases, submitted 
parasites showing parts of all four legs (Fig. 15) of one side, thus 
allowing the diagnosis of arachniasis from microscopic sections alone. 
They are present in large numbers, almost every section containing at 
least part of one and generally several, and lie for the most part within 
air sacs, less often in bronchi. They are surrounded by no special 


WEIDMAN—ARACHNOID PARASITE IN LUNGS 83 


grade of either acute or chronic tissue reaction such as was found in 
cases of Wellman and Wherry (1910). : 

It is impossible to state how common the infestation is in these 
animals because so few come to autopsy. The beasts rarely die upon 
the surface, doubtless seeking seclusion under. ground when they 
become sick, there to remain until they die. As a result, only two 
have come to autopsy in the last eleven years in spite of the numbers 
always on exhibition, and of these, only the one showed pulmonary 
parasites. 

The material’ for the determination of the new species was obtained 
by finely teasing a small portion of the lungs, yielding some fifty or 
more fully developed specimens and no larvae or ova. Of these, the 
females were more numerous than the males in the proportion of two 
to one. They had been fixed in formaldehyd, preserved nine years in 
alcohol (70 per cent.), and were examined after clearing by glycerin 
or Farrant’s medium. From the proportion of tissue examined to the 
total lung substance the lungs must have contained from one to several 
thousand parasites. | 

THE FEMALE 


Females selected at random measured as follows: 


Pubescent Females Ovigerous Females. 
0.170 x 0.087 mm. 0.200 x 0.120 mm. 
0.185 x 0.085 mm. 0.193 x 0.102 mm. 
0.175 x 0.087 mm. 0.185 x 0.105 mm. 
0.180 x 0.090 mm. 0.190 x 0.109 mm. 
0.170 x 0.100 mm. 0.195 x 0.103 mm. 


A female (Fig. 1) observed laterally measured 0.190 mm. in length 
and 0.094 mm. dorsoventrally. 

The body is broadly oval, not quite twice as long as broad, not con- 
stricted, and broadest at about the middle. 

It bears a dorsal shield of subtriangular form with broadly rounded 
angles whose base lies at a line between Coxa II anteriorly and apex 
almost at anus, posteriorly. In the latter region the boundary is not 
sharply marked because the confines pass so gradually into the general 
integument, but laterally it is, the shield curving ventrally here for a 
short distance over the lateral parietes. Anteriorly, it shows sharp 
separation from the anterior part of dorsum in but few specimens, 
notably the immature ones. With mature specimens its anterior border 
appears as an anteriorly directed, more or less gentle slope, and when 
this is very gentle the dorsum may appear to be covered by a sub- 
rhomboidal instead of a subtriangular shield with rounded angles. It 
is possible that these different appearances come about in the following 
way. It will be observed from the measurements given above that the 
mature females are of plumper form than the immature (Figs. 2 


84 THE JOURNAL OF PARASITOLOGY 


and 3), doubtless brought about by the development of the reproduc- 
tive organs which produce an internal pressure resulting in expansion 
of the parietes. Now’'the anterior margin of the dorsal shield is indi- 
cated not by a ridge, nor by special difference in character of integu- 
ments (the integument is of the same character over the whole body), 
but by a downward curved slope (Fig. 1), the steepness of which will 
be more and more reduced as the foot becomes elevated by the internal 
pressure. The progressive reduction of this steepness will, now, with 
maturation, obscure the anterior margin and consequently give more 
and more continuity between the shield posteriorly and the anterior 
dorsal integument. 

The dorsum carries but one pair of hairs: long, and projecting from 
its anterior part close to its lateral margins at a level between Coxae I 
and II. Dorsal pits are large, inconstant, and observed in but few 
(four out of fifty) specimens. Of these the dorsal shield carries two 
rows at the level of Coxa III; a more anterior one consisting of two 
pairs of well outlined pits and an inconstant lateral one, and a more 
posterior row of but one pair, one pit on each side. The anterior por- 
tion of the dorsum shows seven, arranged in two irregular longitudinal 
rows (Figs. 2 and 3). 

There are no eyes. The epistome is rounded anteriorly and largely 
covers the mouth parts as viewed dorsally. Laterally, it curves ven- 
trally to become continuous with the hypostome, with which it forms 
a large tube holding the mouth parts. The hypostome shows two deep 
lateral scallops, with intervening median peak extending well anteriorly. 

Mouth parts project but slightly beyond the body, being deeply 
retracted, the palpi and chelicers so closely compacted as to make their 
morphology indeterminate. It cannot be stated how many joints the 
former have, but it is most probable that they bear no hairs since some, 
if present, would project and be recognized in the many specimens 
studied. In rare cases the mandibles are recognized with untoothed 
chelicers (Figs. 1 and 4). 

The dorsal and contiguous lateral integuments are covered by very 
fine, refractile, granular elevations and show no special linear markings. | 

The venter is divided, like that of Sarcoptes scabiei, into several 
irregular triangular areas by shallow furrows which are probably of 
the nature of synarthroses. They begin near the body middle, radiat- 
ing laterally and curving dorsally, some nearly to the dorsal shield and 
all ending close to one of the coxae. (For details see Figs. 1 and 5.) 
The integument here, as with the dorsum, is covered by extremely fine 
refractile elevations of variable size, shows no linear markings, and 
appears to be of a soft leathery rather than chitinous nature. 


PLATE 1 


EXPLANATION OF PLATE 1 


Fig. 1—Lateral view of female. 
Fig. 2—Dorsum of pubescent female. 
Fig. 3—Dorsum of ovigerous female. Coxae of Legs I hidden. 


Fig. 4.—The rostrum, seen ventro-laterally. Dotted lines indicate the chelate 
mandible seen at a lower level through overlying parts. Mandible folded in 
joints toward mid-line. 


Fig. 5—Venter of ovigerous female. 

Fig. 6—Tarsus of Legs I and IJ. The seta is foreshortened. 
Fig. 7—Tarsus of Leg III. 

Fig. 8—Tarsus of Leg IV. 

Fig. 9—Venter of male. 

Fig. 10—Male external parts. 


EXPLANATION OF PLATE 2 


Fig. 11—Pubescent female. From teasings. 


Fig. 12—Ovigerous female. Also from teasings. The rounded, lighter and 
darker bodies are red blood cells, the darker material lung fragments. 


Fig. 13—Male. From teasings. 


Fig. 14—Very low-power view of microscopic section of lung showing dila- 
tation of bronchus at b (bronchiectasis), irregular distention of some air sacs, 
and broncho-pneumonic exudate in others. 


Fig. 15—Very high-power view of microscopical section of lung showing 
longitudinal section of ovigerous female. Note distention of air sacs—emphy- 
sema. 


Fig. 16—Low-power view of microscopic section of lung showing parasite 
(Pp) in lumen of bronchus (br). 


Fig. 17—Transverse section through parasite in lung. at, alimentary tract; 
ovd, oviduct; ex, inflammatory exudate in lung. Each subdivision of scale 
equals 10 microns. 


PLATE. 2 


11 | 190m 


WEIDMAN—ARACHNOID PARASITE IN LUNGS 85 


' The first pair of legs liés close to but not fused with the capitulum. 
There is a broad interval between it and the second pair, and a broader 
one between it and the last two pairs. The latter are separated from ° 
each other by about the same distance as the first two. As fixed, most 
of the,specimens show the first pair directed anteriorly and the other 


three strongly flexed upon the coxe and directed iaterally and anteri- 
orly, as shown in Figure 2. With many, however, the legs are flexed 
toward the middle of venter (Fig. 1), in which case the iatter gen- 
erally pouts, hill-like, strongly ventrally. The first pair is the shortest, 


measuring 0.07 mm.;' the other three but little longer, measuring 


0.08 mm., and none of the four is as long as the body width. Each is 
composed of five segments, distinctly more slender in the last two legs 
than the first two, particularly the tarsus of Leg IV, which is at least 
four times as long as broad. 

Only the tarsus bears special cuticular appendages. In the case of 
all four legs it bears the following: First, a long hair passing from a 
point well short of its distal extremity; second, three or four? minute, 
short, terminal spurs, the median one or two mucronate and a little in 
advance of the others; third, a small, delicate caroncle extending from 
the base of the median terminal spur. With Legs I and II only, a 
short, stout, rod-like seta (sense-hair) extends in addition from a point 
near its base (Figs. 3 and 6). Legs III and IV do not bear this 
structure. 

The anus is terminal, at junction of dorsum and venter, and sur- 
rounded by no special appendages. Stigmal plates in spite of careful 
search of the abundant material, could not be found. 

The vulvar orifice is ventral, median, fissural, longitudinal, lies at 
level of Coxa III, and ends anteriorly at the arthrosis which passes 
transversely at about the body middle. No ova were observed free. 
A specimen in the body of a female measured 80 by 65y. 

The only internal organ recognizable through tiie cuticle is part of 
the alimentary canal, the approximate position of which is indicated 
by thickly placed, black or brownish-black, fine to coarse granules prob- 
ably consisting of altered blood pigmentary material (hemoglobin ?) 
which has been ingested as food. It appears over a variable area 
extending through a greater part of the body width under the anterior 


1. All measurements from an ovigerous female 0.200 mm. long unless other- 
wise noted. 

2. The borders of the tarsus become membranaceous at the extremity and 
appear to be capable, either naturally or artificially, of being folded centrally. 
This may superimpose the central mucronate spurs so that they appear one, 
but in numerous cases this possibility can surely be ruled out. We are working 
here under very high magnification (142 in. oil immersion lens) where it is 
often difficult to translate optical appearances into positive statements. 


86 THE JOURNAL OF PARASITOLOGY 


third of the dorsal shield, and at times for a short distance beyond it 
anteriorly, or farther posteriorly, on each side. ‘Transverse sections 
of the parasite in lung show two lateral longitudinal tubes hugging the 
dorsum closely which are evidently intestine, and a single larger ven- 
tral one which is probable the oviduct (Fig. 17). 


THE MALE 


The male differs from the female only in its slightly smaller size, 
minor differences in configuration of the posterior ventral synarthrosis, 
and its special genital organs. The measurements of several selected 
at random follow: 


0.175 x 0.102 mm. (Fig. 8) 
0.162 x 0.109 mm. 
0.170 x 0.100 mm. 
0.160 x 0.105 mm. 
0.155 x 0.108 mm. 


The genital orifice lies ventrally, median, at the level of Coxa III, 
is transverse, small as compared to its female counterpart. and bor- 
dered by a narrow chitinous rim. In most cases the copulatory appa- 
ratus projects from it in an anterior direction in the form of a short 
heavy cylindrical piece with rounded ends. The parts are so homogene- 
ous that finer details cannot be asserted with certainty, but it appears 
as though the piece were a tube whose wall is split longitudinally 
through its whole length anteriorly, and whose lumen contains two 
extremely delicate, curved, sharp-pointed spicules (Figs. 9 and 10). 
For differences in configuration of posterior ventral synarthrosis com- 
pare Figures 5 and 8, a female and male. 


PATHOGENICITY 


Since knowledge of the clinical course of the disease is lacking, 
postmortem findings furnish of course the only basis for judging the 
part played by the parasite in producing death. By studying these, it 
was found that all of the lesions above described were acute ones, 
best seen in the microscopic sections, where among other changes 
emphysema and brenchiectasis were described, both of which are com- 
' monly produced by severe coughing. 

Now, these two changes may be caused by either acute or chronic ° 
coughs. In those cases where the cough is chronic, lasting, say, several 
months, it is found that fibrous overgrowths occur in addition, par- 
ticularly in the walls of bronchi, and that infiltrates of lymphocytes 
are also sometimes associated. But none of these are seen in this case. 
Bronchial walls are uniformly thin, and free of cells other than those 
which can be explained by the nearby acute inflammation. There 


va! 


WEIDMAN—ARACHNOID’ PARASITE IN LUNGS 87 


exists here the acute forms of bronchiectasis (or bronchiolectasis). and 
emphysema (so-called). 


It has been already noted that no ova or larva were found in the 
abundant material studied. It may be added that there is no important 
difference in size between the mature specimens. These observations, 
together with the lack of chronic pulmonary tissue changes, lead to 
the belief that the mites were present but a short time, certainly not 
long enough to reproduce, and so probably not longer than a few 
weeks, as Sarcoptes scabiet matures from the larva in about three 
weeks. Under these circumstances it is the reasonable thing to believe 
that it is the parasites which have excited the acute bronchopneumonia 
and so induced death. 


The two cases of Wellman and Wherry concerned parasites of 
squirrels which were well encapsulated in tubercular nodes, i. e., had 
been present for some time. It is possible that the squirrels, too, had 
suffered from an acute attack of bronchopneumonia at the time of 
infestation, and if this be true for them, it should also be thought that 
the disease produced by C. penrosei may also be recovered from at 
times and the mites encapsulated. 


The original source and mode of entry are only speculative, as 
discussed in the case of the monkey infestation which has been 
referred to earlier (1915) as occurring in these gardens. 


ZOOLOGICAL. POSITICN 


Following Banks’ (1905) key, this is indicated as follows: Class, 
Arachnoidea ; order, Acarina; superfamily, Sarcoptoidea; family, Cyto- 
leichidae. He describes the family as consisting of two species, 
Cytoleichus (formerly Cytodites) nudus, and Laminosioptes cysticola, 
and gives family diagnosis as “In skin and cellular tissue of birds. 
Vulva longitudinal.” He does not include in this family the original 
type species, C. sarcoptoides,-Megnin, 1879, perhaps on account of the 
scope of his paper, or perhaps because the species has been later placed 
in another genus. Nor does he give the generic diagnosis of Cyto- 
leichus. which I assume to be still extant, as reproduced as follows 
from Megnin (1879) : 

“Body large, orbicular, convex above, plane below, continued ante- 
riorly by a mobile, inclined, conical, tubular rostrum covered above at 
its base only by an epistome provided with no appendages like joints, 
etc. Legs conical, robust, arranged in two groups, a cephalothoracic 
and an abdominal, the first only being marginal, the epimerae of the 
first pair alone fused to form a-sternal plate, the others free; tarsi 
without terminal hooks, only a ventral simple ambulacrum with cylin- 


88 THE JOURNAL OF PARASITOLOGY 


drical pedicle; the tarsus of the second pair shows at all ages in both 
sexes a blunt cirrus directed above and outward. Ovo-viviparous 
acarians. Type species, C. sarcoptoides. Habitat, air sacs of birds 
(pheasants ).” 


Comparison of the above diagnosis with C. peurosei shows several 
important differences, in spite of which the writer has placed this 
parasite in the genus Cytoleichus, mainly because it resembles the mites 
placed there by Wellman and Wherry (1910) as C. banksi. They were 
found in large numbers in the lungs of two California ground squirrels 
(Otospermophilus beecheyit), each within a tubercle, and occurred 
both on and within the lung substance. Their description is a brief 
one, and so far as it goes agrees fairly with this prairie dog species, 
but their illustration while a simple one, is of value here in that it 
shows (1) the joints of the last pair of legs distinctly heavier than in 
the prairie dog species, and (2) the intestinal markings far posterior. 
The type specimen is not available for original reference since it is 
recorded as in the collection of Creighton Wellman, who cannot be 
located in spite of some correspondence. From the data at hand, 
C. banksi would seem to differ from C. penrosei mainly in that (1) 
the joints of the posterior legs are thicker; (2) it bears no short sense- 
hairs upon the first two pairs of legs, or (3) longer ones upon the 
dorsum, and (4) no dorsal shield is mentioned. It is not possible that 
the two prominent dorsal hairs could have been overlooked by Wellman 
and Wherry (1910) had they been present in the squirrel species, nor 
scarcely the sense-hairs on Tarsi I and II; but it is quite possible that 
the dorsal shield might not be considered an entity by some observers. 
These differences determine a new species, C. penroset.* 


The writer feels that C. banksi and penrosei collectively show wide 
enough differences from the type to warrant the construction of a new 
genus to include them. Thus, C. sarcoptoides, the type, lives in birds, 
the other two in rodents; C. sarcoptoides measures nearly three times 
as large (0.570 mm. by 0.440 mm.), and does not bear the long tarsal 
seta common to C. banksi and penroset. He preters, rather, not to 
multiply genera, but to leave this to some systematist who will study 
a larger group of species than the occasional medical writer. 

C. penroset noy. spec. Specific diagnosis: Grossly invisible, broadly 
oval, with dorsal shield and bearing one pair of long hairs anteriorly. 
Legs nearly equal in length, none longer than body width, each with 
five joints, the tarsus of each with long hair near and delicate caroncle 
at tip. Tarsi of Legs I and II with short stout sense-hair near base 


3. Dedicated to Dr. Charles B. Penrose, the president of the Philadelphia 
Zoological Society. 


WEIDMAN—ARACHNOID PARASITE IN LUNGS 89 


in addition. Wulva median, longitudinal, fissural, between Coxa III. 
Male genital orifice in similar position, but transverse. Females aver- 
age 0.193 by 0.108 mm., males, 0.164 by 0.105 mm. 

Habitat, lungs of prairie dog, Cynomys ludovicianus. 

Type specimen in Philadelphia Zoological Gardens. Autopsy 
No. 1044. 


LITERATURE CITED 


Banks, N. A. 1905. A Treatise on the Acarina, or Mites. Proc. U. S. Nat. 
Mus., 28: 1-109. 

Megnin, P. 1879. Memoire sur les acariens parasites du tissu cellulaire et 
des bourses aeriennes chez les oiseaux. Jour. anat. et physiol., 15: 123-153; 2 pl. 

1895. Les parasites articules, p. 153. 

Weidman, F. D. 1915. Pneumonyssus foxi nov. sp., an Arachnoid Para- 
sitic in the Lung of a Monkey (Macacus rhesus). Jour. Parasit., 2: 27-45. 

1915a. An Arachnoid (Pneumotuber macaci Landeis and Hoepke?) Para- 
sitic in the Lungs of a Monkey (Macacus rhesus). Jour. Comp. Path. and 
Therap., 28: 326-330. 

Wellman, C., and Wherry, W. B. 1910. Some New Internal Parasites of 
the California Ground Squirrel (Otospermophilus beecheyi). Parasit., 3: 417-422. 


90 THE JOURNAL OF PARASITOLOGY 


BOOK REVIEW 


MEDICAL AND VETERINARY ENtToMoLocy. William B. Herms. A textbook for 
use in schools and colleges as well as a handbook for the use of physicians, 
veterinarians and public health officials. New York: The Macmillan Company, 
1915. xii +393 pages. 228 figures. $4.00. 


There have been a number of textbooks published for the use of students 
and others dealing with arthropods and the transmission of disease during the 
past five or six years. The present volume differs from the most of these in 
that it introduces the subject with a concise description of parasitism and the 
morphology of the parts that have to do with transmission. This includes a 
statement as to the classes of parasites, the effect and origin of parasitism, and 
a tabulation with figure§ of the systematic position of animal parasites. There 
is a brief discussion of the internal anatomy, classification and metamorphosis 
of insects, with good illustrations of the various types. It is unfortunate that 
the author has introduced new names for the three types, as there are too many 
already. In practically all the insects that have to do with the transmission of 
disease, the mouth-parts are fitted for piercing or sucking. The derivation of 
this type from the simpler biting type is shown in detail, not only for those 
groups that are known to suck blood, but for all others. This is supplemented 
by a list of the orders arranged according to their type of mouth, together with 
a statement as to their type of metamorphosis. Such a treatment is to be com- 
mended, for the sucking type of mouth is distinctive in form and structure in 
each group where it occurs. The following sixteen chapters discuss fully: how 
insects cause and carry disease, direct and indirect infection, external and inter- 
nal parasites; the life-history, habits and relation to disease of common house- 
hold insects; the biting and sucking lice infesting domestic animals and man; 
the life-history, habits and relation to disease of the bedbug and cone-nose; 
the transmission of malaria, yellow fever and other diseases by mosquitoes, their 
habits, life-history and control; buffalo-gnats and their relation to pellagra; the 
house-fly and its relation to the transmission of intestinal diseases, together 
with measures for its control; the blood-sucking muscids and their relation to 
sleeping sickness and poliomyelitis ; myiasis and the bot-flies; fleas and the trans- 
mission of bubonic plague; ticks and tick-borne diseases; mites as skin para- 
sites; and the venom of bees, wasps, spiders and scorpions. There are also 
included analytical tables for the identification of adult mosquitoes, the families 
of the dipterous larvae producing myiasis, and the families and some of the 
genera of fleas. 

The book can be recommended for its figures, most of them new, and for its 
carefully prepared, well-balanced subject-matter. 


NOTE 
Special courses in parasitology with emphasis upon field and experimental 
work are announced by Dr. George R. LaRue in the program of the Michigan 
University Biological Station at Douglas Lake for the summer of 1917. 


The Journal of Parasitology 


Volume 3 MARCH, 1917 Number 3 


ON: THE. SPOROZOON PABRASTIFES,OF THE FISHES, OF 
WOODS HOLE AND VICINITY 


I. FURTHER OBSERVATIONS ON MYXOBOLUS MUSCULI FROM FUNDULUS 


C. W. Haun 


My knowledge of several points relating to the life history, struc- 
ture, and habits of M. musculi as described in a previous paper was 
incomplete. More recent studies have supplied interesting additions to 
and confirmation of previous observations. The new matter relates to 
the method of infection, the pathological effects, the mode of attack, 
the distribution of the disease within the species, and certain obscure 
stages of the life cycle. 


DISTRIBUTION OF THE PARASITE IN NATURE 


Hitherto my observations have been made upon fish that had been 
captive for one or more days. Since a very large proportion of them 
were found to be infected with both bacteria and Myxosporidia, there 
seemed to be good grounds for expecting fish at large to be infected 
in rather large numbers. But this does not seem to be the case. When 
Fundulus are carefully examined immediately after reaching the labo- 
ratory, the number of fish having lesions of any kind are surprisingly 
few. In one catch of one thousand fish, only eleven had pathological 
abnormalities. One of these eleven fish was infected with My.obolus 
musculi, From another catch of one hundred and seventy-five fish, 
Myxosporidia were found only in three fish which had lesions in the 
integument, there being no other fish having injuries. In a third catch 
of sixty-five fish the integument had typical lesions containing the para- 
sites in but two cases. All of these counts were made when the water 
was at a temperature lower than the maximum in the vicinity of 
Woods Hole. The earliest count made was in July of a remarkably 
cold season. The proportion having Myxobolus was 4.4 per cent, 
while those taken in the latter part of the month of August had as low 
as 0.1 per cent infected. 

The temperature is no doubt an important factor. The water of 
‘the indoor aquaria is warmer than that of the ponds and bays where 


92 THE JOURNAL OF PARASITOLOGY 


the fish are commonly caught. This, in part, accounts for the larger 
percentage of infection in fish that have been confined a day or two. 
But there are two other factors. It has already been demonstrated 
(Hahn, 1913:193) that injuries to the integument encourage the 
entrance of the Myxobolus. An examination of the gills of a number 
of Fundulus has recently revealed the fact that M. musculi is far more 
common on the gills of fish that are apparently healthy than it is in the 
integument and muscle. Fish having injuries and confined in aquaria 
are therefore exposed to infection from the gills of a comparatively 
large number of previously infected fish. These facts explain the dis- 
crepancy in the distribution of the parasites as found in captive fish 
and in free fish. 


EXPERIMENTAL TRANSMISSION OF THE M. MUSCULI AND THE 
CONDITIONS FAVORABLE FOR RECOVERY 


In order to confirm the results of previous experiments along this 
line, two experiments were undertaken. Twelve Fundulus were placed 
in one aquarium jar having a capacity of at least 5 gallons and supplied 
with running water. Six fish were put into a second jar for the pur- 
pose of acontrol. The six controls had incisions cut in the integument 
in exactly the same manner as the fish which were inoculated, but a 
sterile scalpel was used. Bits of tissue known to contain the myxo- 
spores of M. musculi were inserted into pockets made with a clean 
scalpel under the scales of the opercle and head of six of the twelve 
fish above mentioned. Similar bits of tissue were inserted into inci- 
sions made in the integument of the remaining six fish so as to be in 
contact with the body muscle. 

By the second day after the operation, all of the eighteen fish were 
still active. The wounds of all had developed into open infected sores, 
due, no doubt, to the bacteria which enter from the water. But there 
was far greater activity in the wounds of the twelve fish which had 
received infected tissue. The adjacent integument was rough, swollen 
and the scales were loosened. In some the flesh was exposed for a 
distance around the incision and a thick layer of white flaky flesh was 
about ready to fall out of the wound. This condition is unmistakably 
due to the destructive work of the Myxosporidia. Those fish which 
had received infection in the head region had more or less inflamma- 
tion in the vicinity of the lesion and in some cases it had spread under 
the jaw and to the opposite side of the head. In one case the roof and 
floor of the mouth were found later to be highly infected with Myxo- 
bolus. This fish and one of the controls died on the second day of the 
experiment. The latter had a bad wound which proved to have 
numerous myxospores. They probably entered the wound from the 


HAHN—SPOROZOON PARASITES 93 


water or found their way in some way from the gills of one of the 
controls. As stated before, recent observations have shown that 
M. musculi is rather common in the gills of fish that show no signs of 
disease. The same conditions apply to a second control which died on 
the third day. The other four controls recovered and lived throughout 
the period of observation. 

By the sixth day five of the inoculated fish died from the effects 
of the Myxobolus. The parasite was found in the infected tissues in 
each case. Altogether, eight of these fish died, three escaped, and after 
twenty-three days the remaining fish had apparently recovered. The 
three that escaped were seriously afflicted when last seen. 

This experiment was repeated with some slight modifications for 
the purpose of gaining more light upon the natural immunity of the 
host. Infected material was introduced under the integument of four 
Fundulus as follows: (1) Fragments of tissue containing myxospores 
were placed under the integument of the operculum; (2) the same 
material was introduced under the integument of another fish on the 
dorsal side just between the eyes; (3) infected material was pushed 
into slits cut into the integument around the mouth; (4) the infected 
tissue was introduced into the flesh on the left side of the body. These 
four fish were given plenty of food and fresh water. They had been 
confined for thirteen days so that it was safe to assume that there 
were no well developed infections at the beginning of the experiment. 
No controls were kept. 

The locus of the infections all developed into conspicuous lesions. 
The fourth fish developed a large open sore, three-fourths of an inch in 
diameter, with white opaque flesh. It died on the sixth day. The 
muscle around the area over which the integument remained unbroken 
was rich in the trophic stages of the Myxobolus, including some propa- 
gative stages. In the tissue used to infect this fish there were few, if 
any trophoblasts of either propagative or multiplicative stages. Myxo- 
spores were very abundant and other propagative stages were probably 
present. It seems likely that the new host was infected by the latter. 
The rapid hypertrophy of the tissues is characteristic of the disease 
and tends to show that the fish has little or no defence when muscle 
tissue is attacked. 

In Fish No. 1 the muscle of the back and sides was involved by 
some means, probably by the spread of the disease to the dorsal side 
of the operculum. Here again a typical lesion was developed and 
resulted fatally. 

The fate of the other two fish was very different. After twenty-six 
days both were alive and their wounds were healing rapidly. At first, 
both these fishes appeared to have wounds sufficiently serious to cause 
their death. But the thin subdermal connective tissue over the skull 


94 THE JOURNAL OF PARASITOLOGY 


either does not conduct the parasites beyond the reach of immunizing 
agents as in the case of the body muscle, or saprophytic bacteria and 
their toxins have not the favorable conditions to poison the host that 
are provided when the infection occurs in body muscle. Inasmuch as 
there is ample evidence that M. musculi does attack epidermis and con- 
nective tissue, one must conclude that in this case either the defense 
of the fish was sufficient to destroy the parasite before it spread to the 
body muscle or that the parasite passed through its trophic stages and 
had become non-virulent. In the fish which received infection through 
the muscles of the lower jaw, there was nothing to limit the spread of 
the virulent stages into muscles where it would be fatal, such as the 
eye muscles. One is therefore inclined to the view that the parasites 
pass into a comparatively inactive condition. This would require a 
very simple explanation, namely, that the trophic stages develop simul- 
taneously into sporogenic stages. Such was doubtless the case with 
most of the parasites in the primary host. In the latter the disease 
never at any time assumed very injurious conditions. Yet I have 
observed cases of infection in the head region which resulted fatally. 
This particular fish lived for over a month after the disease was first 
observed on the middle of the opercle. It did not spread beyond the 
border of the opercle, and when last observed at the end of the season 
the wasted tissues were rapidly regenerating. At the start, myxospore 
and sporoblast stages alone were encountered in large numbers. All 
of the parasites seem to have developed into sporoblasts and eventually 
myxospores so that the host was safe for the season unless the spores 
germinated again. In the two fish mentioned above, the transfer of 
the myxospores to another host apparently supplied the necessary 
stimulus, or there were still a number of trophoblasts of the propa- 
gative cycle. 

The conditions of the recovery in these three cases were chiefly the 
location of the primary infection. Had the fish not been well fed, they 
would doubtless have died, as have many others having infected jaws, 
eyes, opercles, etc. But food alone will not explain their recovery, 
because I had here two and have had at other times many other fish 
with infections in the body muscle which nearly always kill the fish. 

Recovery in the barbel when afflicted with abscesses caused by 
M. pfeifferi, is possible when there is no external lesion or when no 
vital organ is involved. Usually these are the conditions when the body 
muscle alone is infected. According to de Drouin de Bouville (1908), 
phagocytosis then prevents the fatal accumulation of atrophied tissue. 
As has been already observed, the conditions are just the contrary in 
the Fundulus. When M. musculi invades the body muscle it is rarely 
checked and when the attack is superficial as in the head region, the 
chances of recovery are good. In this conclusion I have assumed that 


HAHN—SPOROZOON PARASITES 95 


the myxospores of M. musculi are not capable of germinating in the 
tissues where they have matured. Mercier (1906) has established this 
as a frequent method of multiplication in M. pfeifferi of the barbel. 
Altogether, the evidence that the myxospores of M. musculi may 
germinate in the original host is negative. The fact that numerous 
myxospores were observed unaccompanied by other stages for such a 
long period in the case above mentioned is, in itself, a sufficient proof 
that, in this case at least, the necessary stimulus for germination of the 
myxospore was lacking. 

In regard to the propagation of M. musculi from fish to fish, it 
may eventually prove that the myxospores may enter the tissues both 
through lesions as is indicated by the above experiments, and through 
the gills and through the digestive tube. Since it has been shown that 
M. pfeifferi is taken into the barbel with its food, the latter mode of 
infection for M. musculi seems the more probable, especially when it 
is recalled that the relations of both parasites to their host are so very 
similar. The attack upon the muscle fibers is almost identical in the 
two species. 

The myxospores of M. inequalis which causes the disease known as 
carp pox, are also transmitted to new hosts by means of the food 
(Wierzejsky, 1898). 

Contrary to my expectation, there is absolutely no evidence that 
Fundulus ever suffers from an internal infection by M. musculi, unless 
it be about the mouth and gill region. 

In the summer of 1915 I again inoculated fish with Myxosporidia. 
In these experiments the ultimate object was to discover if the species 
of Myxobolus hitherto commonly encountered in Fundulus heteroclitus 
would grow and produce the same typical pathological conditions in 
F. majalis and F. diaphanus, and to see if the parasite could be 
recovered in the same host in one of its characteristic stages. 

A Fundulus heroclitus which proved by examination of stained 
tissues to have typical large schizonts in considerable numbers was first 
secured. From two typical Myxobolus lesions in the lateral region of 
the body, bits of flesh about 1 by 3 mm. in size were removed by means 
of sharp sterilized forceps. The subjects were confined in clean 
aquaria, with running sea water for F. majalis and F. heteroclitus, 
and fresh water for F. diaphanus. They were fed regularly each day. 
Inasmuch as it has been shown that lesions free from Myxosporidia in 
fish which are well cared for rapidly recover, no controls were pro- 
vided. This was partly due to the fact that one cannot be sure that 
the water is free from Myxosporidia, since the gills of many Fundulus 
may be infected and presumably disseminate the germ. 

The results of operations upon thirteen fish are summarized in the 
following table: 


sholownu JOU s}UOZTIYOsS 


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98 THE WOURNAL: OF PARASITOLOGY 


The right-hand column of the above table indicates the kind and 
number of Myxobolus in the hypertrophied tissues, especially muscle, 
of the operated fish. In twelve out of thirteen fish the Myxobolus was 
recovered after being introduced. In all cases it had multiplied and 
was growing in a perfectly normal way. There is no evidence that the 
change of host has modified the usual course of the life cycle. 

Considering the last two columns together, one may conclude that 
the parasite encountered a favorable medium for growth in all three of 
the species concerned. In F. diaphanus there is a marked difference 
in the abundance of the Myxobolus as compared with either F. majalis 
or F. heteroclitus. In two cases of F. majalis, one is justified in assum- 
ing that there were large numbers of parasites, though they were not 
actually seen, because in one case the fish died of the disease and the 
slide preparation of its tissues was in some way lost; in the other case 
the extremely degenerate condition of the tissue justifies one in the 
expectation that no parasites will be found. Had the slide included 
muscle near the edge of the lesion, it is certain, on the basis of previous 
observations, that a large number of parasites would have been found. 

One may conclude so far as this experiment goes that F. diaphanus 
is less favorable to the growth and multiplication of the Myxobolus. 
By reference to Columns 6 and 7, it is clear that, notwithstanding the 
smaller number of parasites, the disease is equally if not more destruc- 
tive, having produced extensive necrotic sores and killed all specimens 
of F. diaphanus in three days. The unfortunate failure of the sea 
water at the end of nineteen hours prevented an interesting comparison 
of the endurance of the three species with reference to this parasite. 

These observations prove beyond doubt that there is a succession 
of multiplicative cycles, and that large trophoblasts do not pass directly 
into the propagative condition. The propagative stages are distinctive 
and easily recognized both by their habit and staining qualities. It is 
now certain that some considerable multiplication in the multiplicative 
individuals involving several cycles must intervene before the propaga- 
tive trophoblasts are produced. 

The objection may be made that the culture utilized in the above- 
mentioned experiments was not pure, since one fish known as 1097.9 
proved to be afflicted with both Chloromyxum funduli and M. musculi. 
It is necessary to admit that one could not with precision distinguish 
the trophoblasts of the Chloromyxum from those of the Myxobolus 
unless conditions happened to be very favorable. This is not the case, 
however, if either of these parasites are in the propagative cycle. In 
this case all the stages are distinctive for the two genera. There are 
besides this two very good reasons for believing that the fish from 
which these primary cultures were taken did not harbor Chloromyxum 
to the exclusion of Myxobolus: (1) The Fish 1097.9 is the second case 


HAHN—SPOROZOON PARASITES 99 


of Chloromyxum funduli which I have observed in the tissues of many 
hundred infected Fundulus; (2) no recognizable stages of C. funduli 
could be found in the material available in any of the other twelve 
fish mentioned above. One would hardly expect this particular com- 
bination of circumstances which would provide only one example of a 
parasite in the propagative cycle when they usually advance simul- 
taneously from stage to stage, and at the same time that the initial 
infection be of rare occurrence, one which is encountered about one 
time in two hundred. 

The inoculation experiments which follow are of a similar char- 
acter to the above, and give support to and throw additional light upon 
some of the conclusions mentioned above. The purpose, however, was 
to aid in solving two questions which arise from the following circum- 
stances. I have observed slight differences in the size of the myxo- 
spores from the gill and from the flesh of the Fundulus. In the gill 
I have encountered a range of variability in length from 13.4 to 12u, 


Figure 1 Figure 2 Figure 3 


Fig. 1—Cyst from gill filaments of Fundulus containing four myxospores of 
M. musculi. The cytoplasm around the myxospores is unstained. In this par- 
ticular gill there were a number of these cysts. 

Fig. 2—Cyst from gill filaments of Fundulus containing a small number of 
myxospores of M. musculi. A conspicuous granular cyst plasm with definite 
outer wall characterizes this common type of encystment in the gill. 

Fig. 3—Cyst from filaments of Fundulus containing a large number of myxo- 
spores of M. musculi which have been assembled without any evidence of sur- 
rounding cyst plasm. There is, however, a definite limiting membrane. 68 by 67x. 


and in width from 10.4 to 6y. For those seen in the flesh we have 
recorded elsewhere an average length for apparently mature myxo- 
spores of 14.34 and an average thickness of 6.7p. For obviously imma- 
ture myxospores the dimensions average about 12 by 7.5u. The size 
difference is therefore rendered invalid as an evidence of difference by 
the element of age. Another possible specific difference is suggested 
by the occurrence of myxospores both singly and in sporocysts of 
different sizes (Figs. 1, 2, and 3) in the gills, whereas in the flesh they 


100 THE JOURNAL. OF PARASITOLOGY 


are usually isolated in our smear preparations. This difference can 
scarcely be due to the process of making smear preparations, because 
one should at least find the myxospores clustered if not occasionally in 
pseudocysts. It is very probable in view of what follows that the 
myxospores are either mechanically aggregated in the gills or if nor- 
mally so related, they are mechanically distributed by the action of 
muscular contraction. 

In order to finally settle this question of identity it was planned to 
introduce some of the myxospores of the gill, and if it so happened, 
some of their related trophic stages, into the body muscle. If the 
species were not identical, one would expect a marked difference in the 
pathological conditions and general habit of the parasite, if indeed it 
would grow at all. Some entire gill filaments of F. heteroclitus, 1098, 
which contained the myxospores of a Myxobolus in large clusters, 
singly and in sporocytes having four myxospores in each (Fig. 1), and 
large multiplicative or possibly propagative trophoblasts, were intro- 
duced under the integument of a F. heteroclitus 6.5 inches long. In 
four days the infected fish was dying. The mouth was gaping and 
there was an acute inflammation around the mouth and head. A large 
lesion had developed around the incision and the adjacent flesh under 
the unbroken swollen integument was a purulent mass. It was a typical 
myxosporidian wound. The muscle fibers of the fish were abundantly 
infected with numerous small multiplicative trophoblasts, many large 
trophoblasts and also large masses of multinucleated sporoblasts. 

Unfortunately the water with which these fish were supplied was 
exposed to contamination by other infected fish. The head infection 
was doubtless due to direct contamination by handling or to the infected 
water. But I believe the flesh to have received its deep-seated and 
profound infection from the fragment of gill which was introduced. 

The contaminated water made it necessary to repeat the experi- 
ment. As a number of Cyprimodon wariegatus were available it was 
planned to test the possibility that M. lintoni and M. musculi are one 
and the same species (Hahn, 1913: 206). The gill filaments containing 
one or more large pseudocysts composed of apparently mature myxo- 
spores of the genus Myxobolus were removed from F. majalis. After - 
carefully isolating a single filament it was introduced under the integu- 
ment overlying the body muscle. The details of the experiment with 
summary of the observations will be found in Table 2. 

It should first be noted that Fish 1103.1 died in less than a day, 
and thereupon in Column 8 the visible injury is found to be slight. The 
same condition prevails in 1103.3, but in 1103.2, 1103.4, 1103.7, and 
1103.8 no reason can be given for the non-development of a typical 
lesion. 


HAHN—SPOROZOON PARASITES 101 


If one considers Column 8 it is impossible te deny that in some 
cases, at least, typical lesions do develop; but the evidence is not con- 
clusive. The regular occurrence of one or more stages of the parasite 
in the flesh as indicated by Column 9 éertainly forbids the conclusion 
that the myxobolus of the gill will not grow in the flesh. Allowing 
for the fact that one does not always happen to include in a smear 
prepartion Myxosporidia when present, it may be assumed that all 
the tissues reported in Column 9 contain multiplicative trophoblasts. 
No propagative stages were encountered. In those fish that lived forty- 
five hours and seven days were found large trophoblasts and stages 
which I have considered practically mature, i. e., schizonts. This fact 
harmonizes with the assumption that the transplanted myxospores have 
given rise to the new infection. 

When compared with Columns 8 and 9 of Table 1, Columns 8 and 
9 of Table 2 are not strikingly different, especially if one takes into 
consideration the period of development (twenty-two to forty-five 
hours), and a possibly longer time required for a myxospore to ger- 
minate. One must also consider the relative numbers of individual 
parasites represented in a bit of flesh containing hundreds of indi- 
viduals and a bit of gill filament with only one or two pseudocysts like 
that in Figure 3. Obviously far more significance must be attributed 
to the presence of parasites at all, as indicated in Column 9, than is at 
first apparent. Considering the fragile nature and the relative size of 
myxospores which vary at different stages of development, and the dif- 
ference in the nature of pseudocysts which may be either mechanical or 
due to too limited observations, I feel justified in taking the view that 
there is but one species of Myxobolus in Fundulus, and that it can be 
transplanted both by myxospores and trophic stages. 

The case of the identity of M. musculi and M. lintom is more per- 
plexing. Since M. musculi grew readily (see Table 2) in F. diaphanus 
from fresh water, it might be supposed that it would grow more or 
less in the flesh of C. variegatus. If, on the other hand, the growth in 
C. variegatus had produced a typical tumor and the large type of 
myxospore had been recovered (Hahn, 1913: 206) we might find in 
the above observations evidence of the identity of the two species. 
It should be recalled that the M. lintoni, described by Linton (1891) 
and Hahn (1913), produced in all cases a very characteristic dermal 
tumor, which caused the death of the fish, according to Hahn, in a 
period of from two to three days. Such tumors are never encountered 
in the Fundulus, and nothing suggesting them was preduced in the 
Cyprinodon of this experiment. On the other hand, M. musculi pro- 
duces a typical ulcer in every way comparable to that in Fundulus. 

It is worthy of note that, though the number of cases is small, there 
was an apparent difference between the number of parasites found in 


’ 


102 THE JOURNAL OF PARASITOLOGY 


F. heteroclitus and in either F. majalis or C. variegatus. This, together 
with the slight difference in the degree of development of the lesion, 
indicates that the parasite grows more readily in the muscle of 
F. majalis and C. variegatus than in F. heteroclitus. The number and 
maturity of the myxospores introduced must be taken into account. 
The conclusion is therefore not positive. 

The above experiment, of which Table 2 is a summary, furnishes: 
a minor contribution to the life-history of M. musculi. It would 
appear that if a pure myxospore culture were used in the inoculations. 
and if after seven days large schizonts are found in the second host, 
that not more than seven days is required for the parasite to pass from 
sporoplasm to schizont. Reference to Columns 6 and 9 of the table 
shows that such was the case in Fishes 1103.7 and 1103.8. But 
Fish 1103.5 had many large schizonts which must have developed in 
a forty-five-hour period. At least one cycle may therefore be com- 
pleted in forty-five hours, and probably less, since the number of indi- 
vidual parasites in the twenty-two-hour cultures was far greater 
(1103.2) than the number of myxospores introduced. This conclusion 
is not absolutely certain inasmuch as a gill filament containing a pseu- 
docyst might also contain other stages unseen, but it is very improbable 
if the myxospores are ripe. If trophoblasts were present, they were 
not numerous and the time relations above recorded would then apply 
to the period of a cycle starting with a multiplicative spore rather than 
a myxospore. 

The ease with which one can introduce either multiplicative spores 
and trophoblasts or myxospores and probably propagative trophoblasts 
into the tissues of a healthy fish provides a very plausible explanation 
of the way by which fish whose integument has been broken: may pick 
up the Myxobolus from the water. Thus, though commonly in the gill 
and head region where it is comparatively harmless, it comes to react on 
the body muscle where it is oftentimes fatal. Rough handling and 
close confinement in aquaria tend to provide the ideal conditions for 
the infection of the muscle. 

A final solution of a question which contronted the writer during 
the first stages of his investigations of M. musculi (Hahn 1913: 199), 
namely the possible causative relation of certain bacilli to the patho- 
logical changes in F. muscult,is found in the inoculation experiments. 
When one can produce at will the typical condition by the use of 
Myxobolus myxospores but fails to get it by laceration, one may con- 
clude that the bacteria are purely secondary. Moreover, numerous 
preparations show that the vanguard of the infection is always a tissue 
comparatively free from bacteria. I am not prepared to say that 
bacteria do not poison and kill the host as secondary agents. They are 
probably saprophytic and the primary Myxosporidian parasite prepares 
the way for them. 


HAHN—SPOROZOON PARASITES 103 


SUMMARY OF RESULTS OBTAINED IN INOCULATICN EXPERIMENTS 


1. M. musculi is communicable in all stages of its life-history. 

2. Many multiplicative cycles are repeated before M. musculi 
passes into its propagative cycle. 

3. The Myxobolus which is very common in the gills, where it is 
seldom destructive, is identical with that which occurs in the flesh. 

4. Infection of lesions in the integument takes place upon the 
entrance of any stage of M. musculi from contaminated water. The 
water is presumably contaminated from the gills. 

5. Transplanted M. musculi may continue for some time in the 
. same cycle in the new host. Or they may pass into the next cycle soon 
after the transfer. 

6. Myxospores germinate when transplanted to another fish and 
produce schizonts in considerable less than one day. 

7. The multiplicative cycle requires less than one day and probably 
takes place many times in this period. 

8. The propagative cycle may be reached in 48 hours. 

9. Recovery from infection with M. musculi is possible if the body 
muscle is not involved and if the fish are fed and supplied with oxygen. 
Eye muscles and possibly other parts of the head are also vital. 

10. Recovery is probably possible if the infection occurs when the 
parasite is in or near the end of the propagative cycle even when the 
body muscle is involved. 

11. Progress of the disease is slow in the integument. 

12. The parasite almost invariably migrates from hypertrophied 
tissues. . 

13. Passage from stage to stage is approximately simultaneous. 

14. Fundulus majalis, heteroclitus, and diaphanus and Cyprinodon 
variegatus are culture media for M. musculi.. C. variegatus is a little 
less favorable for its growth but is perhaps less immune to the toxic 
products evolved in this particular kind of a lesion. 

15. The Myxobolus from the gill of Fundulus is identical to that 
which is common in the flesh. 

16. There is no valid reason as yet to consider M. musculi and 
M. lintoni of the Cyprinodon variegatus as one and the same species. 
On the other hand the bulk of the evidence favors the opposite view. 

17. The associated bacteria are either purely saprophytic or secon- 
dary parasites which gain entrance from the water and find the natural 
resistance of the tissues lowered by the Myxobolus. The latter invade 
the normal tissue and leave the atrophied tissues to the bacteria. 

Observations upon the multiplicative stages of M. musculi will fol- 
low in an early number of THE JOURNAL OF PARASITOLOGY. 


104 THE JOURNAL OF PARASITOLOGY 


REFERENCES CITED 

: de Bouville, R. de Drouin. 1908. Maladie des abcés du Barbeau, My-ro- 
boliasis tumerosa, Bull. soc. sci. Nancy, (3) 9: 525-548; 1 pl. 

Hahn, C. W. 1913. Sporozoon Parasites of Certain Fishes in the Vicinity 
of Woods Hole, Massachusetts. Bull. Bur. Fish., 33: 193-214; 2 pl. 

Linton, E. 1091. Notice of the Occurrence of Protozoan Parasites (Psoro- 
sperms) on Cyprinoid Fishes in Ohio. Bull. U. S. Fish Com., 9: 359-361; 1 pl. 

Mercier, L. 1906. Contribution a l’étude du développement des spores chez 
My-xobolus pfeifferi. C. R. soc. biol., Paris, (12) 9: 763-764. 

Wierzejsky, A. 1898. Ueber Myxosporidien des Karpfens. Bull. internat. 
acad. sci. Cracovie, pp. 129-145. 


NOTES ON THE CERCARIAE OF THE BITTER ROOT 
VALLEY, MONTANA * 


ERNEST CARROLL FAUST 


Before the work of Cort (1915) only isolated accounts of North 
American Cercariae had been published. Most of these lacked the 
detail necessary to distinguish species of the same groups which bear 
a superficial resemblance to one another. The general idea which 
Charles Sedgwick Minot voiced is just as true for larval trematodes 
as for any other larvae, when he said “It is not true that embryos are 
alike ; on the contrary, they show class, ordinal, and generic differences 
from one another.’’ This has been the case with the trematode forms 
that have come under the observation and consideration of the writer. 
Some details of difference are visible from a study of the living 
animals. Other points of differentiation require a specific, altho not 
unusual technic. 

During a two-years residence at Missoula, Montana, the writer 
became acquainted with the fauna of the Bitter Root Valley. One of 
the striking features of its fauna is the small number of species, altho 
the number of individuals of each species is large. In contrast with 
this fact is the large number of parasites found in the aquatic fauna of 
the region. Thru the courtesy of Mr. Bryant Walker of Detroit, 
Michigan, who has identified shells from some fifteen collections, the 
writer has ascertained that the gasteropod fauna of the vicinity con- 
sists of three species, Lymnaea proxima Lea, Physa gyrina Say, and 
Planorbis trivolis Say. Thirteen species of cercariae have been secured 
from these snails, species embracing three groups of Digenea. In addi- 
tion, a fourteenth species, a Diplostomulum, has been found in the 
squawfish, Ptychocheilus oregonensis Richardson. Of these fourteen, 
two are Monostomata, two are Holostomata, and the remaining ten 
belong to the Distomata. The writer wishes to take this opportunity 
to thank Professor Henry B. Ward for many valuable suggestions, and 
to express his gratitude to Mr. Norbert Sager for faithful collection 
of material during the summer and fall of 1916. 

Collections were made along the Bitter Root River for a stretch of 
some fifty miles from Hamilton to the confluence of the Bitter Root 
and Missoula Rivers. One collection came from Rattlesnake Creek, 
Missoula. From this latter collection, made in November, 1916, were 


* Contributions from the Zoological Laboratory of the University of Illinois 
under the Direction of Henry B. Ward, No. 80. 


106 THE JOURNAL OF PARASITOLOGY 


secured one species common to several localities along the Bitter Root 
stream, and one new species not found in the Bitter Root Valley. 

All of the species examined have proved to be new. Preliminary 
studies have been made on living specimens to note the general behavior 
of the organism and to work out the details of the excretory system. 
Other organs and tissues have come out more advantageously from 
preserved and stained mounts. Extremely satisfactory results have 
been obtained from material fixed in Gilson’s fluid and stained with 
Delafield’s and Ehrlich’s acid hematoxylin. 


MONOSTOMATA 


Three monostome cercariae previously described for North America 
are: Cercaria hyalocauda Haldeman 1842; C. (Glenocercaria) lucania 
(Leidy) 1877; and C. urbanensis Cort 1914. Two new species are 
contributed from the Bitter Root Valley, Cercaria pellucida and C. 
konadensis. 

CERCARIA PELLUCIDA nov. spec. 
[Figure 1] 

A light infection of Cercaria pellucida was found in the liver tissue 
of Physa gyrina Say, collected near Fort Missoula. A much heavier 
infection of the same species was secured from Lymnaea proxima 
Lea at Corvallis, Montana. The cercariae have an average length of 
0.4 to 0.7 mm., and a width of 0.18 to 0.2 mm. The tail measures 
0.5 mm. in length by 0.07 mm. at the base. The anterior end of the 
body tends to be bluntly fusiform, while the posterior part is elongate 
ovoid, with two symmetrically placed projections where the pair of 
locomotor pockets extend posteriad and ventrad. The pigmentation is 
prominent, centering around the two paired, lateral eye-spots, and a 
third which is single and median. The pigmentation proceeds posteriad 
along six lines, two lateral, two dorsal and two ventral. A single sucker 
at the anterior extremity is aided in locomotion by the paired lateral 
pockets, situated at the posterior extremity of the body somewhat 
lateral to the junction of trunk and tail. 

The animal has a typical “measuring worm” movement, produced 
by the action of the oral sucker and the locomotor pockets, accom- - 
panied by alternate contraction of longitudinal and transverse muscles. 
The “turbine movement” of the tail also pushes the animal forward 
thru the medium. The body of the cercaria is smooth, covered with 
a heavy integument, and the inside of the pharynx has a spinose lining. 

The cercaria is produced in a redia with a tough integument and 
well developed musculature. The redia measures 2.2 by 0.5 mm., and 
has a long single gut-pouch 2.0 by 0.3 mm. The gut is filled with 
reddish orange pigment caused by the digestive action of the organ on 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 107 


the liver tissue of the host. The pharynx is strongly muscular and 
possesses a four-lobed evertible prepharynx with a spinose covering. 
The rhythmic driving of this organ against any object with which it 
comes in contact constitutes the most characteristic movement of the 
redia. The germinal. epithelium is situated in the posterior part of 
the redia. 

The cercaria usually remains in the redia until it is mature and 
ready to seek a new host. A slight pressure on the redia causes it to 
burst at the anterior end, and the rent allows the mature cercariae to 
escape. They do not remain long in the surrounding medium (water 
or liver tissue) before encystment. This process is extremely rapid. 
A mucoid is first poured out around the wriggling worm and within 
this a granular area which acts as a liquid cushion for the cercaria. 
During the process of encystment the cercaria has coiled upon itself 
so that the resulting cyst is spherical. Encystment is so rapid that the 
‘tail is not dropped until the completion of the cyst. 

Cercaria pellucida is characterized superficially by no special feature 
that would differentiate it from any of the larger species of monostome 
cercariae with three pigment eye-spots. Internal characters, however, 
readily distinguish this form from previously described species. The 
locomotor pockets are smooth internally, differentiating the species from 
C. imbricata Looss (Looss, 1896) and C. ephemera Nitzsch (Ssinitzin, 
1905). The tail has no central gland cells, so that the common median 
excretory tube is surrounded by the ordinary parenchyma cells. The 
excretory system of the trunk consists of a complete circuit with the 
anterior extremity just posterior to the median pigment eye, and the 
posterior limit of the system in the bladder. When contracted, the 
bladder is superficially triangular with the excretory pore caudad. The 
system is filled with many large granules of a high refractive index. 
They are probably of a derived protein nature and disappear on appli- 
cation of strong acids and alkalies. However, after treatment with a 
non-acid fixing agent (mercuric chlorid), they are acid-resistant. 

The eye-spots consist of a terminal ganglion cell for each spot, 
surrounded by a pigment cup. The eyes open dorso-laterad. They 
have a direct connection with the brain. 

Most specific of all the systems are the genital organs of Cercaria 
pellucida. The germaria (ovary and two testes) are situated in the 
posterior reaches of the trunk just anterior to the excretory vesicle. 
The ovary is median; the minute testes are lateral to the ovary. No 
Laurer’s canal has been found. The vitellaria consist of five pairs of 
glands within the confines of the excretory circuit and three pairs 
situated more laterad. They cover a considerable area of the ventral 
surface. The glands are filled with fine granules closely associated. 


108 THE JOURNAL OF PARASITOLOGY 


The ducts from the glands traverse the region on each side intermediate 
between the inner and outer series, and meet one another in the region 
of the ovary, emptying thru a common duct into the ootype. From this 
region there proceeds forward along a median line the long slender 
uterus, which enlarges just behind the median eye to form the vagina. 
To the left of the uterus is the common vas deferens which has resulted 
from the junction of the vasa efferentia just anteriad to the ovary. 
It runs parallel to the uterus, and at its anterior end expands into a 
cirrus pouch. 

The body is crowded with cystogenous gland cells filled with rhab- 
ditiform granules. Crowded in between these cells are the parenchyma 
cells and connective tissue. The cystogenous cells with their contents 
give to the worm a milky translucent appearance. 

The digestive ceca are given off from the esophagus just behind 
the plane of the paired eye-spots and extend to the posterior extremity 
of the body. Their lumina are filled with a jelly in which are granular 
inclusions. 

CERCARIA KONADENSIS nov. spec. 
[Figure 2] 

From the same individuals of Lymnaea proxima Lea at Corvallis, 
Montana, from which Cercaria pellucida was obtained, there were 
found in lesser numbers specimens of another new species of mono- 
stome cercariae for which I propose the name Cercaria konadensis, the 
specific designation of which is derived from the Indian term for 
Bitter Root. 

This species is very unlike Cercaria pellucida. It is considerably 
smaller, having a length of 0.4 to 0.46, and a width of 0.1 to 0.16 mm. 
The tail is equally long and has a transverse diameter of 0.03 to 
0.04 mm. at the base. The anterior end is lanceolate, as is also the 
extremity of the tail. Superficially, the most striking feature of this 
cercaria is the lack of-the median pigment eye, accompanied by a lesser 
amount of pigmentation in the posterior half of the body. This stands 
in contrast with the trioculate monostome cercariae where the pigmen- 
tation is more extensive. In this respect Cercaria konadensis bears 
similarity to C. urbanesis Cort (Cort, 1915), which has only two true 
eyes and a median condensation of pigment, and contrasts with 
C. ephemera Nitzsch (Ssinitzin, 1905; Lebour, 1905), and C. imbricata 
Looss (Looss, 1896). 

The germinal epithelium of the redia in which the cercaria develops 
is modified into a central rachis which is proliferated from the pos- 
terior part of the body. This redia is much smaller than that of the 
trioculate cercariae, with a length measurement of 1.7 mm., and a 
diameter in cross section of 0.35 mm. The pharynx is comparably 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 109 


smaller and the gut extends only about three-fifths the way to the 
posterior extremity. It is about 1.0 mm. long and about 0.1 mm. in 
diameter. There is no oral armature. The walls of the redia are 
muscular and fairly well covered with integument. 

Superficially, Cercaria konadensis cannot be distinguished from 
C. urbanensis Cort, altho the writer believes it is on the average more 
attenuate than that species. On the other hand, there are internal 
characters that readily allow a differentiation. 

The posterior locomotor pockets are lined on their inner faces with 
a group of ten to twelve gland cells which probably pour out a secre- 
tion for attachment of these organs to the surface of the contact 
objects. There exist six paired groups of glands in the tail, lying just 
laterad to the caudal excretory tube. Each group constitutes a rachis, 
with the broadened end of ‘mature cells directed toward the trunk, and 
the acute end of proliferating cells directed distad. This paired series 
of twelve groups of glands in the tail constitutes the readiest mark of 
distinction for the species. It separates. it from C. pellucida, on the 
one hand, and on the other from C. urbanensis Cort, in which the 
writer has found six pairs of glands in the tail, each gland composed 
of a single polygonal cell. The exact number of these cells is not 
stated by Cort (1915), altho he mentions their presence. 

The excretory system is not dissimilar on the whole to that of other 
monostome species. The bladder is small, 14 to 15 thru the longi- 
tudinal axis of the larva and 16 to 17 in breadth. The excretory tubes 
empty into the bladder from the extreme antero-lateral angles. The 
general aspect of the vesicle is a strongly compressed spheroid. The 
excretory pore is dorsal. ' 

The genital fundaments are hardly as clearly outlined in Cercaria 
konadensis as they are in C. pellucida. The ovary is just anterior to 
the excretory bladder, is pyriform in shape and lies dorsad to the 
ootype. It has a distinct Laurer’s canal. The testes are slightly 
postero-lateral to the ovary. The cells of these glands are poorly 
defined, altho the efferent duct of each is indicated by a double row 
of cells. These ducts lead into a common efferent duct anterior to the 
ovary, and this runs forward to the right of the uterus, ending in a 
bulbous cirrus pouch a slight distance behind the vagina. As is usual 
for monostome cercariae, the yolk glands consist of a paired inner 
series of five glands anda paired outer series of three glands. They 
are very diffuse, dendritic, and are readily traced to the common lateral 
ducts which proceed posteriad, and, finally, turning mesad in the plane 
of the ovary, empty thru a common vitelline duct into the ootype. The 
uterus reaches from the region of the ootype to the plane of the 
anterior vitelline glands, where it enlarges into the vagina. This loca- 


110 THE JOURNAL OF . PARASTTOLOGY 


tion of the genital atrium is considerably behind the two eye-spots, so 
that this feature distinguishes it from that of C. pellucida. 

The eye-spots are two in number situated superficially to the right 
and left of the roots of the posterior dorsal nerve trunks from which 
they receive innervation. Pigmentation is centered around the brain 
and its immediate nerve trunks. 

Cystogenous glands fill the greater portion of the connective tissue 
complex. Encystment is rapid. This species is precocious in that it 
encysts frequently within the tissues of the snail. 


HOLOSTOMATA 


Few Holostomes, either in larval or adult condition, have been 
described for North America. Diplostomum cuticula, D. grande, 
D. volvens, and Tetracotyle .typica, all Old World forms, have been 
reported for North America, and Stafford (1904) has described a new 
species, Diplostomum parvulum. The descriptions of these forms are 
not detailed, and it is doubtful if they are sufficient for exact deter- 
mination of the species. An undescribed hemistome larva occurring in 
the vicinity of Urbana, Illinois, has features common to all of these, 
and especially characteristic for Diplostomum cuticula; yet it is 
undoubtedly a new species, as determined by internal structure. An 
isolated case of a holostome larva yet unnamed has been recorded by 
Rettger (1897). 

The writer has found two holostomate larvae in the Bitter Root 
fauna, one a representative of the Hemistomes, and one a representa- 
tive of the Holostomes. 


CERCARIA PTYCHOCHEILUS Ov. spec. 
[Figure 3] 

This species was found in very large numbers (several thousand) 
in the agamic stage in the mesentery of Ptychocheilus oregonensis 
Richardson, caught at Stevensville and Carlton, Montana, in April, 
1915. As a hemistome larva encysted in a vertebrate, it is technically 
a Diplostomulum. ‘The stage in the mollusc has not been secured. The 
larvae were included in large vesicular cysts of a translucent consis- 
tency. The cyst was attached to the mesentery by a disc. Upon 
transfer to normal saline or Ringer’s solution the end of the cyst was 
split and the larva emerged. 

Cercaria ptychocheilus is conspicuous because of its abbreviated 
posterior portion and its elongate patelliform anterior region. The 
body averages from 0.48 to 0.63 mm. in length by 0.17 to 0.37 mm. in 
width. There is a medium-sized pharynx. The ceca extend to the 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 111 


acetabular region. The oral sucker is small and the acetabulum some- 
what larger. 

The excretory system consists of a bellows-shaped bladder into 
which leads a single median excretory trunk. In the mid-acetabular 
region it divides into three trunks, one brench proceeding forward 
and one each directed laterad. The lateral trunks form anastomoses 
both anteriad and posteriad. The median trunk continues its course 
unbranched until it approaches the region of the forking of the diges- 
tive tract, where it branches. The branches bend laterad right and left, 
and join the anterior anastomoses of the lateral trunks. Thus the 
system is bisymmetrical and constitutes a double circuit for the con- 
duction of the excretory products. All of the tubules are filled with 
granules. 

The genital system is typically holostomate, with the genital pore 
posterior. A muscular organ anterior to the acetabulum represents the 
original genital pore, which has lost its connection with the genitalia. 
The ovary is a club-shaped organ lying transversely posterior to the 
acetabulum and continuous mesad with the oviduct on the left side. 
The vitellaria are diffuse, ventro-lateral. No uterus is present in the 
larva. Two testes are situated on the right side, ventral and posterior 
to the ovary. No vasa efferentia or vas deferens is present. The 
genital pouch lies ventral to the excretory bladder. It is muscular and 
has paired groups of glands emptying into it. 


CERCARIA FLABELLIFORMIS 710UV. spec. 
[Figure 4] 

This holostome larva possesses a right and left sucking disc in 
addition to the oral and ventral suckers; in consequence it belongs to 
the group designated as tetracotyle larvae. The species was found in 
the livers of a large percentage of Physa gyrina Say from three collec- 
tions in the vicinity of Corvallis, Montana, in October, 1916. Some of 
the parasites were encysted, others were free in the liver tissue, and 
still others were in the redia. No mature cercariae were found in the 
rediae. The animal is broadly spatulate from ventral aspect. The 
length of the mature cercaria is 0.48 to 0.56 mm., and the width, 
0.44 mm. The redia measures about 0.5 mm. in length and 0.16 mm. 
in width. The rhabdocoel gut is short. A pharynx is present. The 
birth-pore is situated on the ventral side, slightly lateral. Within the 
redia the germ balls are developed from the germinal epithelium 
localized at the posterior end. These balls develop into other rediae 
or tetracotyle larvae, both within the same redia. 

Characteristic of the younger Cercaria flabelliformis is the tetra- 
cotyle suctorial apparatus, consisting of the oral and ventral suckers 


112 THE JOURNAL OF PARASITOLOGY 


and two lateral sucking disks. Behind the acetabulum are two trans- 
versely plicated lappets. The lateral sucking disks are modified as the 
larva matures so that they become lappets and come to lie within a cup- 
shaped hollow. Even at an early stage the larva is encysted. 

The excretory system consists of a very truncate common vesicle 
and two long vesicular tubes. In the region of the transverse lappets 
these trunks give off a transverse tube which joins the two lateral 
systems. On its anterior side are given off tubules in fan-shaped 
arrangement. Lateral to the transverse trunk, and extending posteriad, 
are numerous anastomoses. 

The genital cell-masses bespeak a typical holostome system. The 
yolk glands consist of paired tubular chords extending from the fork- 
ing of the gut to the testes. They have large vesicular cells. Thick 
ducts lead into the. ootype which is ventral to the ovary. The uterus - 
leads obliquely from the right side of the ovary posteriad into the 
genital pouch. The testes are large pyriform glands, lying at the sides 
of the genital pouch. They open into the cone near the genital pore. 


DISTOMATA 


Distome cercariae may be grouped according to certain larval char- 
acters, which, altho not holding over to the adult trematode, are 
coexistent with other characters that are more deep seated. The Bitter 
Root species of the distome larvae consist of six xiphidiocercariae, two 
echinostome cercariae, and two furcocercariae. Among the xiphidio- 
cercariae were found six species. 


CERCARIA CRENATA 20V. Spec. 
[Figures 5 and 10] 


A heavy infection of this species was found in 13.6 per cent. of 
Lymnaea proxima Lea collected at the springs at Fort Missoula, Mon- 
tana, in October, 1916. It is a minute larva, oblong-ovate in contour. 
_ The length of the trunk is 0.25 mm. and the width 0.13 mm. A weak 
tail, 0.15 to 0.16 mm. in length by 0.02 to 0.03 mm. in cross section at 
the base, is inserted into an aspinose caudal pocket, just posterior to 
the excretory vesicle. The larva possesses a very acute stylet fastened 
into the dorsal roof of the oral sucker, about 30u long and 5y broad at 
the base (Fig. 10). 

The cercaria develops from the germ balls proliferated from the 
localized germinal epithelium within the very simple oval sporocyst. 
The mature sporocyst measures about 0.5 by 0.25 mm. It is non- 
muscular and has no organs of attachment. It depends for movement 
on the movement of the cercariae developing within it. 

The prominent muscular parts of the larva are the large oral sucker, 
60 in diameter, the small acetabulum, 30u in diameter, the small but 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 113 


powerful pharynx with @ median transverse constriction, and the 
crenate muscular excretory bladder. 

Above the vesicle the excretory trunks diverge as a U from a single 
stem, each arm giving off a posterior and two anterior tubules. 

The digestive system consists of a filiform ‘esophagus and two ceca 
in the form of a typical furculum. The oral pocket anterior to the 
esophagus is large and deep. The pharynx sphincter surrounds the 
posterior half of the esophagus. When at rest the ceca end at the 
posterior margin of the acetabulum. Salivary glands consist of two 
series, an outer group of eight small cells and an inner group of five 
large cells. These groups empty into the oral cavity thru separate 
ducts. 

The genitalia are represented by cell masses in the acetabular and 
postacetabular regions of the body. Antero-sinistral is Laurer’s canal 
and proceeding forward is the coiled uterus-vagina fundament, ending 
in the genital pore mesad and just anteriad to the acetabulum. The 
testes are elongate pyriform bodies, extending postero-laterad at a 40° 
angle. The vitellaria are poorly developed, altho a few follicles and 
three main ducts are visible as they proceed mesad toward the ootype. 
The vitellaria are probably limited in the adult to the third quarter of 
the body. 

CERCARIA GLANDULOSA 10V. Spec. 


[Figures 11 and 16] 


This species was obtained from a heavy in fection of liver tissue of 
Physa gyrina Say from the vicinity of Hamilton, Montana, in October, 
1916. The cercaria is moderately small, with a length of 0.45 mm. 
and a width of 0.2 mm. The tail has a length of 0.35 mm. and is 
0.05 to 0.06 mm. in trans-section at the base. It is set into a caudal 
pocket, with locomotor spines in the lateral pockets. The stylet is 
placed in the roof of the oral sucker. It has a blunt point and measures 
39 in length by about 5 in width (Fig, 11). 

The sporocyst is extraordinarily simple in structure with a delicate 
epidermal wall. It is obovoid and measures 0.34 mm. in long diameter 
by 0.17 mm. in short diameter. The cercaria is proliferated from a 
localized germinal epithelium. 

The cercaria is characterized by an unusual supply of glands. 
Cystogenous glands fairly crowd the other body structures. A paired 
series of nine glands of salivary nature empties into the oral cavity. In 
addition, the entire digestive tract is covered with gland cells, especially 
in the region of the muscular pharynx, so that the alimentary tract 
simulates superficially a cluster of grapes. The esophagus and the 
crura are short, just clasping the anterior margin of the acetabulum. 


114 THE JOURNAL OF PARASITOLOGY 


The oral sucker is somewhat larger than the acetabulum; the 
former measures 86u in diameter and the latter 66y. 

The excretory system consists of a compressed vesicle and two 
cornua, each of which receives a single posterior tube and a single 
anterior tube. The anterior tube has three tributaries in the region of 
the acetabulum. Posterior to this region it receives several transverse 
tributaries (Fig. 16). 

The genitalia are typically Plagiorchid. The ovary is situated 
dorsal to the acetabulum and merges into a large uterus-vagina funda- 
ment. Laurer’s canal is prominent, arising from the vicinity of the 
ovary and turning dorso-sinistrad. The testes are not distinguishable 
at this time. The vitelline follicles extend from the extreme oral 
region to the extreme posterior region. Vitelline ducts run mesad 
toward the region of the ovary. 


CERCARIA DIAPHANA 10V. spec. 
[Figures 12 and 17] 


The species Cercaria diaphana is a delicate larva of such a beautiful 
gray as to remind one of a mere shadow. It is extremely transparent. 
It occurred as a heavy infection in the liver tissues of Lymnaea 
proxima Lea obtained from the Bitter Root River, Corvallis, Montana, 
in October, 1916. When contracted the larva is compressed ovoid, and 
measures 0.2 to 0.26 mm. in length and 0.1 to 0.12 mm. in width, but 
it is capable of extraordinary expansion. The tail is lanceolate, 
0.15 mm. in length by 0.04 mm. in trans-section at the base, where it is 
included within the spinose caudal pocket. 

The sporocyst in which the cercaria develops, is oblong, measuring 
0.35 by 0.15 mm. One end may be drawn out as a sort of club-shaped 
process. An extremely simple germinal epithelium produces the cer- 
cariae. It is non-localized and lines the whole body cavity. No 
external organs of attachment or movement are present. 

The oral sucker of Cercaria diaphana measures 44, in cross section, 
and the acetabulum only 32y. The tail is deeply sunken at the base 
into the posterior caudal pocket. There are a few (eight to ten) long 
spines at the dorsal edges of the pocket. A unique stylet is located in 
the dorsal roof of the oral cavity. It measures 39u in length by 5y in 
breadth at the base. Its anterior reinforcement is confined to two 
dorso-lateral plates at the anterior end. Between these lies a minute 
spine 5u in length by 0.5 in diameter (Fig. 12). 

The excretory system consists of a highly muscular, compressed 
vesicle, from which there extends anteriad a Jong median protuberance. 
This trunk forks to form two trunks slightly posteriad to the acetab- 
ulum. Just postacetabular each trunk becomes constricted and connects 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 115 


with a common lateral tubile. The tubule receives three main branches, 
two from the cephalic region and one from the caudal portion (Fig. 17). 

The digestive system consists of a long slender esophagus and crura 
of equal length. The latter are broadly furculate. A small muscular 
pharynx is provided with an immense mass of gland cells. The 
pharynx itself measures about 15 in cross section, while the gland 
complex includes a sphere of 65, diameter. In addition, there are the 
paired salivary glands, eight in each paired group, small and poorly 
developed, emptying into the oral pocket. 

The genital cell masses are typically Plagiorchid. Vagina, Laurer’s 
canal and ovary are situated dorsad to the acetabulum. Testes are not 
yet visible. Vitelline follicles extend from the posterior margin of the 
oral hood to the base of the caudal pocket. Ducts arise from the ovary 
posterior and lateral, and are directed antero-mesad. 


CERCARIA DENDRITICA /0V. Spec. 
[Figures 13 and 18] 


The species Cercaria dendritica was obtained from the liver tissues 
of highly infected Lymnaea proxima Lea, collected from the sloughs 
of the Bitter Root River at Fort Missoula, Montana, in October, 1916. 
The larva is an extremely muscular individual, altho the tail is weak 
and of questionable value in movement. The cercaria performs a 
characteristic “measuring worm” movement as it travels forward. It 
is about 0.38 mm. long by 0.15 mm. wide, and has a tail 0.16 by 
0.04 mm. at the base, inserted into a spinose caudal pocket. 

The oral and ventral suckers are large and well developed. They 
measure 62» and 60p, respectively, in diameter. The tail is included at 
its base within a caudal pocket provided with stout spines thruout 
the entire lining. The stylet in the roof of the oral cavity measures 
44u in length by 14 in breadth thru its basal knob. The quill is tri- 
angular, scutate, and is joined to the shaft by a median and a pair of 
lateral reinforcements (Fig. 13). 

The sporocyst is well developed. It consists of an elongate ovoid 
body provided with an oral sucker 80» in diameter and is well supplied 
with muscular elements. The sporocyst itself measures 0.38 by 
0.11 mm. The germ cells are situated at the posterior end. The cer- 
caria is obovate, possibly due in part to the extreme muscular develop- 
ment of the oral sucker. 

The excretory system deserves special emphasis. The sub-spherical 
crenate vesicle is remarkably muscular and the two cornua which are 
anterior are equally muscular. At the extreme anterior reaches of 
each cornu three tubules flow into it, two from the anterior portion 

ed one from the posterior éxtremity. The tubules are dendritic 
(Fig. 18). 


116 THE JOURNAL OF PARASITOLOGY 


The digestive tract consists of a large pharynx 3Qy in transection 
and 36 long, a short esophagus of about two-thirds the length of the 
pharynx, and extremely rudimentary crura, hardly as long as the non- 
muscular portion of the esophagus. Salivary glands, eight in number 
on each side, arise from the region just anterior to the oral cavity. 

The genital organs are well-defined. Ovary and uterus lie on the 
right side over the acetabulum. On the left side is the definitely out- 
lined Laurer’s canal, and just caudad to the acetabulum are the testes. 
Yolk glands consist of a pair of rather slender racemes arranged in 
zigzag fashion all along the lateral reaches of the cercaria, from the 
extreme ends of the trunk. The vitelline ducts lead into the ootype 
from a posterior angle. 

Cystogenous cells fill all of the mesenchyme spaces of the body. 
They are large, white, oval bodies. All of the cercariae reach maturity 
almost synchronously. They are mature when they break thru the 
wall of the sporocyst and swim out into the surrounding medium. The 
tail is soon cast off. In fact, the animal travels much more rapidly 
without the tail than with it, for it can then use the spines of the caudal 
pocket. Encystment is slow; the cyst is a thin oval membrane within 
which the larva is coiled. 


CERCARIA MICROPHARYNX 10V. spec. 
[Figures 14 and 19] 

This species was secured from the liver tissues of Lymnaea proxima 
Lea obtained from the Rattlesnake Creek, Missoula, Montana, in 
November, 1916. The cercaria is oval, minute, measuring 0.18 mm. in 
length by 0.09 mm. in width. The tail is 0.14 mm. long by 0.03 mm. in 
width at the base. It is fairly active. 

Anteriad is the stylet organ, superficially set in the oral roof, so that 
its leverage is poor. The organ is rounded at the point, and reinforced 
all around the margin. Across the top is a thin translucent mucoid 
velum. The stylet is 344 long and 5y in breadth along the shaft 
(Fig. 14). The tail is inserted proximally into the caudal introvert. 
provided with spinose projections. The entire body is covered with 
minute spines arranged in diamond pattern and decreasing in size from 
the anterior to the posterior margin. 

The excretory system is entirely non-muscular. The vesicle is sub- 
spheroid and laterally compressed, and the two cornua which arise 
antero-laterad are likewise sub-spherical. Each receives three tubules, 
a small posterior, a large outer, and a small inner anterior tubule 
(Fig. 19). 

The digestive tract is diminutive. H consists of a minute pharynx 
around the middle portion of the esophagus, and small vesicular crura 


FAUST—CERCARIAE OF BITTER ROOT VALLEY Miy 


Paired groups of salivary glands, each with eight cells in the group, 
are found in the acetabular region. The pre-pharynx is provided with 
a large spheroidal group of small gland cells. 

The genital cell masses consist of a non-differentiated band of 
tissue just dorsal and posterior to the acetabulum, in the neighborhood 
of the future ootype, a uterus-vagina cell mass running cephalad over 
the acetabulum, and in addition broad bands of yolk follicles extending 
along the margins from the pharynx region to the caudal pocket. The 
beginning of the tests are not yet distinguishable. Laurer’s canal is 
definitely set off to the right of the uterus. 

The sporocyst is ovoid, measuring 0.24 by 0.18 mm. It is remark- 
ably simple, with a single layer of epidermal cells constituting the body 
wall. The germinal epithelium is non-localized. There is an inter- 
cellular complex of excretory channels in which are found many excre- 
tory calculi. When the germinal epithelium has been exhausted, the 
cercariae maturing last drop off their tails and encyst within the sporo- 
cyst. The cercaria is provided with many minute subspherical cystog- 
enous cells thruout the parenchyma. 


CERCARIA RACEMOSA 10V spec. 
[Figures 15 and 20] 

This ornate cercaria was found in the liver tissues of Lymnaea- 
proxima Lea obtained from the sloughs at Fort Missoula, Montana, in 
October, 1916. It is oblong-spatulate, with a delicate quill stylet and a 
fluted tail. The body measures 0.29 mm. in length by 0.11 mm. in 
width, while the tail is 0.22 mm. in length by 0.04 mm. in width at the 
base. 

Cercaria racemosa is found developing in rhomboidal sporocysts 
about 0.93 mm. long and 0.56 mm. in trans-section, with a poorly 
defined attachment pocket at one end. At the antipodal end is the 
localized germinal epithelium from which the cercariae develop. The 
cercariae grow to maturity within the sporocyst. 

The body of the cercaria is aspinose. The slender stylet measures 
12u in length by 2u in width at the base (Fig. 15). This is reinforced 
only at the pointed tip. It is advantageously set in the roof of the oral 
cavity so as to give a good leverage. 

A pair of non-pigmented eye-spots are present superficially in the 
region of the brain ganglia. ; 

The excretory organs consist of a truncate vesicle, a median tube 
anterior to the vesicle, and two fusiform cornua which receive racemose 
tubules at their anterior extremities. The vesicle contains two groups 
of three cells each, probably glandular, attached to the anterior margin 
of its inner wall (Fig. 20). 


118 THE SOURNAL OF PARASITOLOGY 


The digestive tract consists of a small muscular pharynx, a long 
slender esophagus, and short crura clasping the anterior margin of the 
acetabulum. Paired salivary glands, eight in each group, are situated 
in the acetabular region. Their long ducts open into the oral cavity. 
The genital cell masses are restricted to the acetabular and post- 
acetabular portion of the cercaria. A vagina and a Laurer’s canal are 
discernible. Vitelline glands are confined to the region just. postero- 
lateral to the ootype. No testes can be made out. 

The tail is of considerable power in swimming and is not readily 
detached. No encystment occurs for some time after the cercaria is 
placed in a watch glass of normal saline solution. 


ECHINOSTOME CERCARIAE 


Echinostome cercariae possess a circum-oral collar- with spines and 
usually contain three large flame cells in the anterior portion of the 
excretory system. The further criteria added by Cort (1915: 37), 
namely, an excretory system opening on each side of the anterior part 
of the tail, and “tail powerful, longer than body,’ may or may not be 
typical of individual species: they are not family characters. Of the 
two species of this family that have come under the writer’s observa- 
tion, only one has a tail longer than the trunk, while neither one has 
the excretory system opening on each side of the anterior part of the 
tail. 

The two species described by Cort (1915) as echinostome cercariae, 
C. trivolvis and C. rubra, with the probable echinostome larva, C. 
reflexae, constitute the only species of this group previously described 
from North America. Two new species are contributed from the Bitter 
Root collection. ; 

CERCARIA TRISOLENATA 10UV. Spec. 
[Figure 6] 

This species is an echinostome larva of unusual features. It is con- 
siderably more slender than the usual species in this group. The tail is 
short and lanceolate. The acetabulum is studded with spines. The body 
is 0.45 mm. long and 0.1 mm. wide when the animal is at rest. The tail 
measures about 0.2 mm. in length and 0.016 mm. in section at the base. 
The acetabulum is a third larger in diameter than the oral sucker which 
measures 30u. Around the dorsal margin of the collar and extending 
a short distance ventrad is a ring of spines, 36 in number, in a single. 
altho somewhat irregular line. These spines are aciculate, yet blunt at 
the base and at the extreme tip. 

The cercaria is developed in rediae found in the liver tissues of two 
snails, Physa gyrina Say and Planorbis trivolvis Say, collected along 
the entire course of the Bitter Root River. It is one of the two dis- 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 119 


tinctly cosmopolitan species of the valley. While the infection of the 
Physa was heavy (22 to 100 per cent of all Physas examined) and the 
Planorbis infection was 50 per cent, the infection of the individual 
Planorbis was much heavier than that of the individual Physa. 

The redia when mature measures 1.0 mm. in length by 0.25 mm. in 
cross-section. It is provided with a small pharynx, 55y in trans-section, 
and a large rhakdocoel gut extending the entire length of the body 
cavity. The locomotor “feet” occupy a position about one-third the 
body distance from the oral opening. Proliferation of germ balls 
occurs from the posterior end. The rhythmic movement of the redia 
is due to its own muscular action and that of the daughter cercariae. 

The excretory system of the cercaria consists of a small obtruncate 
bladder and the lateral canals which remain unbranched until they 
reach the cephalic region. Here each forms a single deltoid anastomosis 
and end in three flame cells. The tubules are filled with excretory 
granules. The caudal tube is single, median, and unbranched thruout. 
the entire course. 

The digestive tract consists of a long esophagus with a small 
pharynx mid-way along its length, and a pair of long crura extending 
posteriad to the subterminal region. Soon after the crura arise from 
the esophagus they cross under the excretory trunks and run parallel 
to them externally all the way posteriad. 

The genitalia are not well developed in the larva. They consist of 
an ovarian mass some distance behind the acetabulum, a vagina to the 
right and just anterior to the acetabulum, and two testes, one behind 
the other in the posterior extremity of the trunk. 

Encystment starts with the rejection of the tail and later the slow 
formation of a semi-membranous cyst capsule from the abundance of 
glandular material with which the cercaria is filled. The cyst is very 
transparent, but extremely resistant to mechanical and chemical dis- 
turbances. The trisolenate arrangement of the excretory tubules 
cephalad is clearly seen thru the cyst membrane. 


CERCARIA BIFLEXA 10V. Spec. 
[Figure 7] 

This form is broadly wedge-shaped at the cephalic margin and 
rounded posteriad, with long powerful tail, large groups of salivary 
glands and marked bodily activity; the cercaria is extraordinarily 
destructive to the host which harbors it. It was found in a small per- 
centage of Physa gyrina Say from the vicinity of the Buckhouse Bridge 
near Fort Missoula in November, 1916. 

The collar spines are elongate-ovoid, 10» in length, 42 in number. 
The acetabulum is situated in the posterior third of the trunk. It is 
60 and the oral sucker 50 in diameter. The body measures 0.45 to 


126 THE JOURNAL OF PARASITOLOGY 


EXPLANATION OF PLATE 


Fig. 1—Dorsal view of Cercaria pellucida; specimen partially contracted, 
showing eye-spots, excretory and genital systems. 80. 


Fig. 2—Dorsal view of Cercaria konadensis; specimen relaxed, showing eye- 
spots and anterior pigmentation, excretory and genital systems, and gland cells 
of tail. >< 105; 


Fig. 3—Ventral view of Cercaria ptychocheilus; specimen freed from cyst, 
showing digestive, excretory and genital systems. 80. 


Fig. 4—Ventral view of Cercaria flabelliformis; young specimen within 
cyst, showing digestive ceca, excretory system and lateral suctorial cups. 50. 


Fig. 5——Dorsal view of Cercaria crenata; digestive, excretory and genital 
systems shown; salivary glands in two series, inner and outer, empty into oral 
cavity thru long ducts; cystogenous cells not shown. > 170. 


Fig. 6.—Ventral view of Cercaria trisolenata; digestive and excretory sys- 
tems shown.  X 150. 


Fig. 7—Ventral view of Cercaria biflexra; excretory and genital systems 
shown. 105. 


Fig. 8.—Posterior two-thirds of Cercaria gracillima; specimen shows genital 
cell masses; testicular follicles proliferated from the posterior end. 270. 


Fig. 9—Dorsal view of Cercaria tuberistoma; excretory system and salivary 
glands shown. 170. 


Fig. 10—Stylet organ of C. crenata. X 540. 

Fig. 11—Stylet organ of C. glandulosa. 370. e 
Fig. 12—Stylet organ of C. diaphana. 540. 

Fig. 13.—Stylet organ of C. dendritica. 250. 

Fig. 14—Stylet organ of C. micropharynx. « 540. 
Fig. 15.—Stylet organ of C. racemosa. 333. 

Fig. 16.—Excretory vesicle of C. glandulosa. > 75. 
Fig. 17—Excretory vesicle of C. diaphana. 170. 

Fig. 18—Excretory vesicle of C. dendritica. > 113. 
Fig. 19—Excretory vesicle of C. micropharynx. 270. 
Fig. 20.—Excretory vesicle of C. racemosa. 150. 


Reference line in Figs. 1-9 and 16-20, 50 long; in Figs. 10-15, 10 long. 
Lines in Fig. 6 indicate important regions not discussed in this paper. 


PLATE 


SSS ee - 
SAE O FI! N e5 
es ea N 
rete 


om Cy le per: 

WaiN Reeye re ee 
epee ph ° Si eye 
pelea rot Peru cad pd 


See Sy opr 
“a Ree atria 


y 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 121 


0.5 mm. in length by 0.13 to O15 mm. in width. The tail is of equal 
length to the body and 0.06 mm. in cross-section at the base. 

The redia of Cercaria biflexa measures 0.4 mm. in length and 
0.09 mm. in cross section. The locomotor “feet” are found in the 
posterior third of the body. The pharynx is moderately large, 40 in 
cross section, and well developed. On the other hand, the rhabdocoel 
gut is short, extending only thru the cephalic fourth of the body. The 
posterior margin is characterized by a number of small integumentary 
spines. Cercariae are produced from a localized germinal epithelium 
in the posterior part of the body. 

The excretory system of the cercaria consists of an elongate vesicle 
and a U-shaped trunk system leading into it anteriorly. Lateral tribu- 
taries are received by these two branches thruout the body tissues. At 
the anterior end cephalad to the collar prominence, the main tube on 
each side becomes attenuated, loops back on itself as far as the collar . 
region, then turns again anteriad and ends in three flame cells. The 
excretory tube in the tail is single, two-fifths of the distance distad. 
There it forks, altho the bifurcations never open laterad. 

The digestive tract consists of a very long esophagus, extending to 
the acetabulum, and a pair of ceca arising just preacetabulad and 
extending nearly to the posterior margin of the body. An inner and 
an outer series of salivary glands, fifty to sixty in each series, occupies 
the larger part of the body ventral to the excretory trunks. They 
empty thru united lateral ducts into the oral cavity. 

The genital cell masses are fairly well developed in the cercaria. 
An ovary posterior to the acetabulum, vitelline ducts and a uterine 
duct have a common center at the ootype. The uterus proceeds antero- 
dextrad around the acetabulum and ends in a large muscular vagina 
anterior to the acetabulum. Two testes are observable posteriad to the 
vitelline ducts, one almost on top of the other. The animal encysts 
readily. While no encystment was noticed within the redia, it may take 
place as soon as the cercaria escapes from the mother. Most of the 
specimens were found encysted in the tissue of the host. 


THE FURCOCERCARIAE 


This group of larval trematodes is characterized by a forked tail 
and, as far as the writer knows, the absence of a true pharynx. How- 
ever, glands in the pharyngeal region may lead one to consider the 
mass a pharynx, which is evidently the error Looss (1896) has made 
in his study of Cercaria vivax Sons. The apharyngeal furcocercariae 
are undoubtedly larval Schistosomidae, as demonstrated by the experi- 
mental work of Leiper (1916) and by a close comparative study which 
the writer has made on larvae and adults. Two new furocercous larvae 


122 THE JOURNAL OF PARASITOLOGY 


have been obtained from the Bitter Root Valley. These, in addition to 
Cercaria douthitti (Cort, 1915), constitute the only described forms of 
North American Schistosome larvae. 


CERCARIA GRACILLIMA 10U. spec. 
[Figure 8] 

This is an extremely slender tho wiry individual. It has a body 
length of 0.13 to 0.16 mm. and a width of 0.02 to 0.03 mm. The tail is 
approximately twice as long as the body and is equally divided between 
the simple and bifurcate portions. This cercaria is of common occur- 
rence in the Bitter Root Valley, altho it is most abundant in the lower 
part of the valley. It was found abundantly in liver tissues of Physa 
gyrina Say, and in Lymnaea proxima Lea, along with a large infection 
of Cercaria micropharynx. 

The body is provided with an oral sucker covered with spines; the 
ventral sucker measures about 12. The oral sucker can be drawn into 
the esophagus. Vestiges of non-pigment eye-spots are found dorsally 
in close proximity to the brain. 

The cercariae develop in sporocysts from a localized germinal 
epithelium. The proximal end is provided with an attachment disk. 
The sporocyst is about 0.5 mm. long at maturity and 0.025 mm. wide. 
It has no musculature and depends on the cercariae within for its 
motility. The cercariae escape thru a rent in the wall of the sporocyst. 

The excretory system includes a common non-muscular vesicle at 
the posterior margin of the trunk, and two lateral canals which anas- 
tomose frequently and characteristically in the anterior two-thirds of 
the body. Flame cilia are present in a restricted region of the main 
tubes in the posterior third of the body. The junction of body and 
tail is accompanied by an “eyelet anastomosis,” commonly found in 
furcocercous larvae. The common tube of the anterior unbranched 
region of the tail, branches into the rami of the tail. 

The digestive system consists of a long esophagus which branches 
to form the ceca just anteriad to the acetabulum. The ceca end at the 
posterior margin of the acetabulum. Paired salivary glands, four to 
each series, lying in the posterior third of the body, open by long ducts 
into the oral cavity. 

Anterior to the acetabulum are the ovary-uterus cell mass on the 
right and that of the cirrus on the left. Posterior is the male germinal 
epithelium from which is proliferated a large number of testicular fol- 
licles. Ventro-lateral are lines of vitelline glands which empty their 
products thru ducts into the ootype anterior to the acetabulum. 

Iencystment has not been noted in the species. 


FAUST—CERCARIAE OF BITTER ROOT VALLEY 123 


CERCARIA TUBERISTOMA nov. spec. 
[Figure 9] 

Two prominent tubercles are present at the anterior end of the 
spineless body of this species. The chamber for the oral sucker occu- 
pies the core of the anterior third of the worm. The body is about 
0.2 mm. long by 0.05 to 0.06 mm. wide. The tail is about 0.32 mm. 
long, of which the unbranched portion constitutes approximately one- 
half. It measures 35 at the base. The ventral sucker measures 30. 
The larva was found in Physa gyrima Say at Corvallis, Montana, in 
October, 1916. The infection was light. 

The cercaria develops in sporocysts, which are about 0.5 mm. long 
and 0.05 mm. in trans-section. At one end is a sucking disk, and at the 
other end is the broad attachment organ. The germinal epithelium is 
localized at this latter end. 

The excretory system consists of a small muscular maliform bladder 
situated posteriad, and slender lateral trunks which receive occasional 
branches more anteriad. No flame cell areas have been made out. The 
“eyelet anastomosis” at the junction of the body and tail is muscular. 
A slender median caudal canal divaricates just anterior to the bifurca- 
tion of the tail. At the proximal end of the tail are given off a pair of 
lateral tubules which are recoiled on themselves. 

The digestive system is of the usual type for the furcocercariae. 
The genital anlagen have not been worked. Encystment has not been 
observed in the species. 


REFERENCES CITED 


Cort, W. W. 1915. Some North American Larval Trematodes. Ill. Biol. 
Monogr., 1: 447-532; 8 pls. 

Lebour, ‘Marie V. 1907. Larval Trematodes of the Northumberland Coast. 
Trans. Nat. Hist. Soc. Northumberland, n. s., 1: 437-54; 5 pls. 

Leiper, R. T. 1916. On the Relation Between Terminal-Spined and Lateral- 
Spined Eggs of Bilharzia. Brit. Med. Jour, 141% 

Rettger, L. J. 1897. Some Additions to Our Knowledge of the Anatomy 
and Embryology of the Holostomidae. Proc. Ind. Acad. Sci. for 1896: 224-225. 

Ssinitzin, D. Th. 1905. Distomes des poissons et des grenouilles des environs 
de Varsovie. Matériaux pour Vhistoire naturelle des Trématodes. Mém. Soc. 
Nat. Varsovie, sect. biol., 15; 210 pp.; 6 pls. 

Stafford, J. 1904. Trematodes from Canadian Fishes. Zool. Anz. i275 
481-495. 


THE DEVELOPMENT OF GREGARINES AND THEIR 
BELALION: TO THE HOST TISSUES: ay 
IN SEENOPHORA' LACTARIA 
WATSON * 


MinniE Watson KAMM 


The object of this paper is the depiction of the stages through 
which the sporozoite passes in the species Stenophora lactaria Watson 
in becoming a free adult sporont. Whether or not the trophozoite at 
any stage in its development possesses an epimerite and the effect of 
the parasite upon the cell parasitized will also be discussed, conclusions 
reached affecting only the particular species under consideration. This 
effect upon cells parasitized will be reported in several other genera 
before conclusions can be stated as to the general influence upon the 
host-cells. 

Stages in the growth of the parasite from the sporozoite to the 
sporont have been described by many writers. Léger and Duboscq 
have studied the development of Pyxinia (1902), Stylorhynchus (1904), 
and Pileocephalus (1909a), and to some extent of Stenophora (1904) ; 
Laveran and Mesnil of Gregarina (1900) ; and Siedlecki, Brasil, Caul- 
lery and Mesnil, and Hesse of parasites in the tunicates and annelids. 
In very few instances has a complete series of stages been shown. 

To the writer’s knowledge, consecutive stages from the incipient 
penetration to the vacation of the cell by the parasite have not been 
depicted for the genus Stenophora. I am able to offer nine stages, 
somewhat arbitrarily chosen, in the development of a single species, 
the species chosen being Stenophora lactaria from the milliped Callipus 
lactarius (Say), described by the writer (1915: 29-30; 1916; 72-4). 
The intestines of several hosts were removed intact, fixed with corro- 
sive-acetic, and sectioned. Sections were cut 4p thick and stained with 
Ehrlich’s hematoxylin. All the intestines proved heavily infected. The 
lumen reveals parasites in the proventriculus and in the large intestine, 
but intracellular stages are generally lacking except in the first-named 
portion.t In several instances parasites were found boring through 
the walls into the coelom. Successive movements have been traced 
from the penetration of the muscular layer of the digestive tract 


* Contributions from the Zoological Laboratory of the University of Illinois 
under the direction of Henry B. Ward, No. 88. 

* The digestive tract of the milliped (Fig. 12) is, according to Leidy (1853), 
divided into (a) and (b) six salivary glands, (c) esophagus, (d) proventriculus, 
(e) two bile ducts, (f) broad opaque cuticular curtain, (g) ventriculus, (h) 
large intestine, (7) rectum. 


KAMM—DEVELOPMENT OF GREGARINES 125 


through the coelomic epithelial layer until the parasites are free in the 
coelom. This phenomenon was reported by the writer in a former 
paper (1916:29). The gregarine possesses no boring apparatus; it 
has no chitinous style and it even lacks an epimerite ; thus, mechanical 
apparatus being absent, the hypothesis is made that the means used is 
chemical and that the body of the parasite secretes a fluid destructive 
to the cell epithelium. The cells in the immediate vicinity of the tubular 
opening are crowded back and disorganized and their nuclear material 
scattered. Both the nuclear and cytoplasmic protoplasm stain less 
deeply than the normal. This fluid present is either a secretion of the 
parasite available for purposes of penetration or the normal excretion 
of the parasite which is toxic to the adjacent tissues. 

After the cyst has been discarded from a host along with its feces 
and has dehisced, the spores are liberated and accidental parasitism 
occurs, the released spores being eaten by a host of the same or a nearly 
related species,” even by the original host. The spore, upon reaching 
the alimentary canal, loses its sporocyst by the action of the digestive 
juices, releasing, in all the Eugregarinae, eight falciform sporozoites. 

The stages in development from the incipient to the mature para- 
site which have been studied are as follows: 

The liberated sporozoite, which is slightly motile, although it 
possesses neither cilia nor flagella, reaches the epithelium of the pro- 
ventriculus and penetrates between the terminal cilia into the free end 
of one of the absorptive cells (Fig. 1). The sporozoite can be readily 
detected, although less than 5p in length, because of its intense colora- 
tion, staining darker than the cytoplasm of the cells. A nucleus can 
be discerned as it is still darker. The sporozoite loses its falciform 
shape and rounds off at one end, penetrating the cell by the remaining 
pointed extremity. 

It journeys down the cell past the nucleus, the pointed end pre- 
ceding (Fig. 2).* The parasite is small enough to make the passage 
without injuring the cell other than by the temporary crowding of the 
nucleus. 

It comes to rest at the end of the cell, next the muscular layer 
(Figs. 3 and 4, a). The sporozoite now becomes trophic, viz., a 
trophozoite. With the beginning of growth and consequent excretion, 
the parasite effects a chemical change upon the parasitized cell. The 
cytoplasm becomes slightly vacuolated and stains a little less deeply 
than normal, but its nucleus is not yet visibly affected. 

In the next stage, the parasite has lost its characteristic sporozoite 
shape and has become a subspherical body of larger dimensions and 


* Léger and Duboscq (1902) have shown that if the spore is eaten by any 
other animal, it will not dehisce but passes intact through the digestive tract. 

* Léger and Duboscq record that in Stenophora aculeata the sporozoite pushes 
the cell nucleus ahead of it toward the muscular layer gradually absorbing it. 


126 THE JOURNAL OF PARASITOLOGY 


with a conspicuous nucleus containing one karyosome, which is char- 
acteristic of the adult nucleus (Fig. 4, b). The host cell has become 
still further vacuolated and stains still less deeply. The nucleus is now 
affected, for it, too, stains more faintly. 

The trophozoite now becomes differentiated into protomerite and 
deutomerite separated by a septum (Fig. 4, c). The nucleus and 
karyosome have grown more rapidly than the parasite and are pro- 
portionally much larger than before. It is already apparent that the 
protomerite stains more intensely than does the deutomerite. This 
continues into the mature sporont stage. The young trophozoite gen- 
erally orients itself so as to lie “head’’ downward, 1. e., with the pro- 
tomerite next the muscular layer, but exceptions occur. In some 
instances also the parasite lies at right angles to its usual position or 
in the antero-posterior plane of the host. In a few instances out of 
hundreds, the protomerite was seen to be directed toward the lumen. 
Léger and Duboscq record this in Stenophora aculeata (1904), and 
Mercier in Cephaloidophora talitri (1912). The former authors name 
the two possibilities, viz., that the sporozoite penetrated the cell pos- 
terior end first or else turned after entrance. The cytoplasm of’ the 
cell is still further destroyed. It may be vacuolate with the nucleus 
intact and seemingly but little changed (Fig. 5) or vacuolate with the 
nucleus already destroyed, its remaining chromatin massed at the base 
( Hig). 

The protoplasm of the two divisions of the trophozoite is becoming 
differentiated (Fig. 6). In the protomerite it stains deeply and consists 
of small homogeneous granules, while in the deutomerite it is more 
coarsely granular and less closely packed together. The nucleus of the 
parasite has now begun to assume the shape of that in the adult 
sporont; it has become ellipsoidal and is now smaller in proportion to 
the size of the gregarine than it has been before. The protomerite has 
acquired a papillate apex which is retained in the adult. This is the 
only structure in this species which may be compared to an epimerite. 
It is not a true epimerite, for it performs no function. It may possibly 
be a vestigeal remnant from a lower group of gregarines which pos- 
sesses and uses a true epimerite, but the relationship of the families of 
gregarines has been little discussed and the higher or lower position of 
the Stenophoridae is not determined. Some members of the genus 
appear to possess epimerites, as S. nematoides Léger and Duboscq 
(1904) and S. dipolcorpa Watson (1916) ; one species retains a minute 
apical style (S. aculeata Léger and Duboscq (1904) ) ; but most species 
have no trace of an epimerite at any stage of development. 

The parasite has by this time acquired something of the normal 
sporont shape (Fig. 7). It has grown so as to absorb several adjoin- 
ing cells besides that first parasitized, nuclear and cytoplasmic vestiges 


KAMM—DEVELOPMENT OF GREGARINES 127 


remaining at the base at one side of the apex of the protomerite. By 
growth and expansion the animal has laterally compressed the cells 
which border it, leaving a small opening into the lumen of the alimen- 
tary tract. It is my opinion that a part of the parasite’s nourishment is 
acquired by absorption direct from the intestine through this opening. 

The stage shown in Figure 8 is very similar to the last. The space 
leading to the lumen is larger and the contiguous cells have been forced 
farther apart with a consequent compression of many cells, their sub- 
spherical nuclei having become very long and slender. It is to be noted 
that the deutomerite is growing faster than the protomerite and is 
forcing itself down over the former at the septum, while the proto- 
merite is flattened against the muscular layer. 

The trophozoite has become capable of living free in the intestinal 
lumen (Fig. 9). It no longer receives sufficient nourishment from the 
epithelium and through the small opening into the lumen, and is forced 
out henceforth to lead a free existence in the lumen. Just what forces 
it out of the epithelium, I am unable to say. One would not be 
inclined to assume in it the power of volition with the ability to leave 
the cell at the critical moment; but, on the other hand, one cannot 
assume that the cells force it out by swelling and expressing it, for up 
to now they have, been passively forced apart by the growing parasite. 

By whatever means, the animal has left, after absorbing nourish- 
ment from many cells which are not entirely destroyed but only dis- 
torted with their nuclei shrivelled. These cells are probably able to 
revive themselves and acquire new vigor, unlike the first few cells, 
which were totally destroyed. The animal has not straightened itself 
out yet from the cramped position when embedded, and the deuto- 
merite still overlaps the protomerite. The liberated trophozoite has 
become a free living sporont. 

The young sporont (Fig. 10) must now receive all its nourishment 
from liquids in the alimentary tract and not indirectly through the 
media of cells. The animal is rotund in appearance and sluggish; 
the epicyte is seen to be thicker at the septum than elsewhere; the 
papillated apex of the protomerite is apparent, and there is visible 
for the first time a minute indentation in this apex which is frequently 
reported for this genus. 

The fully developed sporont is much more graceful (Fig. 11). 
The deutomerite has grown more rapidly than the protomerite, leav- - 
ing the latter a small conoidal segment while the latter has elongated 
and become slender and tapering. The nucleus acquired its permanent 
shape in an early stage, and now remains elongate-ellipsoidal with 
one large karyosome. 


128 THE JOURNAL OF PARASITOLOGY 


It is seen that in the species considered there is no epimerite. 
Nourishment, then, takes place by absorption from surrounding cells 
directly through the epicyte of the parasite. An epimerite would, 
moreover, be superfluous to an animal which is intracellular in devel- 
opment, useful only in a species in which one end only of the parasite 
is embedded in an epithelial cell. 

The parasitized cell is apparently unaffected until the sporozoite 
has begun to grow. Then the cytoplasm becomes reticulate, vacuo- 
late, and resistant to the staining fluid. It commences to shrivel in 
length and in width; atrophy has set in. This may be due to the toxic 
influence of the parasite, but it is undeniably due in part at least to 
the absorption of its vital fluids by the rapidly growing parasite. No 
hypertrophy is noted from the first. The nucleus is affected less 
readily than the cytoplasm, but soon shows a resistance to the stain. 
For a time the chromosome count is unaltered, although the size of 
the individual chromosomes is reduced. The growing animal utilizes 
the space left by the contracting cell and compresses adjoining cells 
in its proximity, while the cells become elongate and wider at the 
uncompressed ends. The posterior end of the gregarine soon projects 
into the lumen, for it forces the cells so far apart that they are no 
longer able to overlap the end of the parasite. The latter now 
undoubtedly receives some of its nourishment from the lumen and the 
drain on the cells themselves is decreased. This probably accounts 
for the fact that frequently not more than two or three cells are 
actually destroyed, the others being compressed only during the 
occupancy of the parasite. When the latter departs, the cells return 
tc something like their former shape, and the space through which 
the parasite left almost immediately closes over. 

The cells surrounding the parasite always are separated a trifle 
from it, leaving in many instances a thin clear area around it. I think 
this is due to the fact that in fixing the parasite shrinks slightly more 
than does the host tissue, rather than to the fact that the parasite 
affects the cells toxically and causes them to draw back. 

The cells of the proventriculus lie in lobes (Figs. 1, 4, 7, and 9), 
the deep-seated lobes being the ones generally parasitized. Only in 
unusual cases do the outlying ones harbor gregarines. The deeper 
seated cell forms a safe harbor for the young sporozoite where it is 
not in danger of being swept along in the lumen by the undigested 
_food masses and by the animal’s movements. These shorter cells also 
afford easy exit when the parasite is ready to leave the epithelium, 
being about as long as the mature trophozoite when it leaves. 

The trophozoite always lies next the basement muscular layer, but 
not actually in contact with it; it never remains half way up the cell. 
As aforesaid, it is usually placed “head downward.” 


KAMM—DEVELOPMENT OF GREGARINES 129 


That the gregarine is intra-cellular rather than inter-cellular is seen 
from the early stages when the cell itself contains the parasite; with 
later stages alone this could not be determined. 

Several writers have noted that in the species which they studied 
distinct hypertrophy of the parasitized cell occurred. Laveran and 
Mesnil (1900) and Léger and Duboscq (1909a) have shown that the 
parasitized cell decreased in length, but at the same time increased 
abnormally in width due to the influence of toxic excretions of the 
parasite. The latter authors think the cell is not killed, but assumes 
its normal shape and function after being relieved of its burden. 
They state, however, that the influence of the parasite upon the host 
cells is very different in different hosts, with different parasites, and 
even. in different parts of the same host. 

The present research has shown that there is no hypertrophy of 
the cell, but that the originally parasitized cell shrinks from the start 
without widening and is destroyed, and that the two adjoining cells 
are in many instances also destroyed. Other nearby cells are tem- 
porarily compressed and elongated, later to return to their normal 
functions ; their staining reaction is unaffected. 


SUMMARY 


Consecutive stages in the growth of Stenophora lactaria Watson 
are depicted. 

This species does not possess an epimerite. 

Development is intracellular and the parasitized cell is destroyed. 

No hypertrophy is indicated at any stage of development. 


REFERENCES CITED 

Laveran, A., and Mesnil, F. 1900. Sur quelques particularités de l’évolution 
d’une Grégarine et la réaction de la cellule-héte. C. R. soc. biol., 52: 554-7 ; 1 fig. 

Léger, L., and Duboscq, O. 1902. Les Grégarines et Vépithélium intestinal 
chez les trachéates. Arch. parasit., 6: 377-473; 5 pl. 

1904. Nouvelles recherches sur les Grégarines et l’épithélium intestinal des 
trachéates. Arch. Protistenk., 4: 335-83; 2 pl. 

1909. Etudes sur la sexualité chez les Grégarines. Arch. Protistenk., 
17: 19-134; 5 pl. 

1909a. Protistes parasites de l’intestin d’une larve de Ptychoptera et leur 
action sur l’hote. Acad. roy Belg., 1909: 885-902; 4 pl. 


Leidy, J. 1853. A Flora and Fauna within Living Animals. _ Smithson. 
Contrib. Knowl., Wash., v. 5, Art. 2: 1-67; 10 pls. 


Mercier, L. 1912. Cephaloidophora talitri, n. sp., Grégarine parasite du 
Talitre. C. R. soc. biol., 72: 38-9; 1 fig. 


Watson, M. E.. 1915. Some New Gregarine Parasites from Arthropods. 
Jour. Parasit., 2: 27-36; 2 pl. 
1916. Studies on Gregarines. III. Biol. Monogr., v. 2, no. 3, 258 pp.; 15 pl. 


130 THE JOURNAL OF PARASITOLOGY 


EXPLANATION OF PLaTes 1 AND 2 


Fig. 1—Sporozoite (s) beginning to penetrate an epithelial cell of the 
intestine. 

Fig. 2——Sporozoite (s) ascending the cell. 

Fig. 3—Sporozoite (s) at rest at the base of the epithelial cell. 

The line at the right represents 50 u. 


Fig. 4.—Three stages in the growth of the parasite: (a@) same as shown in 
Fig. 3; (b) trophozoite with enlarged nucleus and one karyosome; (c) larger 
trophozoite with formed septum, the cell nucleus destroyed and cytoplasm some- 
what vacuolated. The line at the right represents 50 u. 


Fig. 5.—Still larger trophozoite with cell nucleus yet intact altho the cyto- 
plasm is affected. The line represents 25 y. 


Figs. 6, 7, and 8—Stages in the growth of the trophozoite, cell nuclei in 
each instance destroyed, vestiges remaining at the “head” of the parasite. The 
line represents 50 x. 


Fig. 9—Trophozoite leaving the epithelium and migrating into the intestinal 
lumen. The line represents 50 x. 


Fig. 10—Sporont soon after emerging and still rotund in appearance. 


Fig. 11—Mature sporont from lumen showing proportional growth of 
protomerite and deutomerite. 


Fig. 12—A copy of Leidy’s figure (1853, Plate VII, Fig. 21) of the digestive 
tract of Julus marginatus. 


PLATE 1 


PLATE. 2 


Fee is y 

x of 
fet SA See 
Rater ~~ - 


= 


oe 


THE CERCARIAE OF NATAL 
F. G. CAwsTon 


During the months of April, May, June, and July of 1916, I exam- 
ined 1,500 molluscs from the rivers and fresh-water pools of Natal. 
They included several.species. Limnaea natalensis is a common form 
with a dextral shell. Physopsis africana, a common mollusc amongst 
decomposing vegetation, has a blunt-pointed sinistral shell with a trun- 
cate columella. Planorbis pfeifferi is a common form with a round, 
flat shell. Planorbis leucocheilus is not unlike it, but is much smaller. 
Isidora tropica is fairly common; it has a blunt-pointed sinistral shell. 
Isidora forskali is rarer and has a conical shell. In one brickfield I 
found a large number of specimens of Ancylus (ferrissia) burnupi, 
which has a small oval shell. 

Two hundred eleven specimens harbored cercariae of various kinds. 
Infected specimens were most common in one brickfield at Durban 
and in a small pool along the course of the Umsindusi River at Pieter- 
maritzburg, which had been formed as a result of an overflow of the 
river. Infected specimens were most frequently met with in May and 
June. All of the cercariae possessed a long, slender tail; those that 
were found in specimens of Physopsis africana more often than not 
possessed divided tails. The tail was easily detached from the body of 
a cercaria and continued to move for some time after becoming free. 
All of the cercariae were distomes; the oral sucker was terminal, and 
in a few specimens the posterior border of the sucker was incomplete ; 
the acetabulum was situated slightly nearer the tail end of the body. 
None of the cercariae had spines or stylets, and there were no projec- 
tions from the body or tail. A pharynx was noted in only one form, 
and eye-spots were present only in cercariae from one specimen. 


CERCARIA CATENATA 


A large cercaria, Cercaria catenata, from the Toll Gate brickfield 
at Durban, present in about 30 per cent of Planorbis pfeifferi and 
in fewer Limnaea natalensis and Physopsis africana developed in 
rediae. These rediae gave an orange color to the liver-substance of 
infected specimens. The rediae were very mobile and possessed two 
pairs of locomotor appendages. The posterior extremity was pointed. 
There were infoldings of outer cuticula at the extremities of the pos- 
terior extremity of the redia and of its appendages which were not 
unlike suckers. The rediae contained a somewhat distended intestine, 


132 THE JOURNAL OF PARASITOLOGY 


a large amount of orange pigment, and several fully developed 
cercariae. On one occasion a cercaria was seen attempting to draw 
itself through the ruptured wall of the redia by means of its suckers. 
The head of the cercaria varied in appearance, but was often shaped 
like a leaf. It possessed a large oral sucker and a large acetabulum or 
ventral sucker; a chain of blackish granules, varying in number from 
about twenty-five to twenty-eight, lay on each side of the divided 
alimentary canal. The tail of the cercaria was as long or slightly longer 
than the body and tapered towards its extremity.. Hypodermic injection 
of a large number of both rediae and cercariae into a guinea-pig threw 
no light whatever on the life history of this cercaria, which was the 
only form found to develop in rediae. 


““TADPOLE CERCARIAE 


Sixteen sporocysts, containing leptocercous distomes, or “tadpole” 
cercariae, were found in Physopsis africana from the Umsindusi. 
Similar sporocysts were present in two specimens of Limnaea natalen- 
sis from the same source. The sporocyst intersected almost the entire 
liver-substance of infected specimens, giving it a whitish appearance. 
The cercariae consisted of a body with two suckers and a tail which 
was about the same length as the body. There was a divided gut and 
an elementary bladder. Some forms presented a stumpy appearance, 
others were longer. Some of these cercariae found in Durban sug- 
gested the appearance of Schistosome cercariae in every respect except 
that their tail was not divided. 

No cercariae were found in specimens of Isidora (over fifty were 
examined from infected places), and furcocercous cercariae were 
found only in specimens of Physopsis africana. Some specimens of 
this latter mollusc harbored more than one form of cercaria. Occa- 
sionally one came across a specimen which harbored both the “tadpole” 
and furcocercous forms. 

Ninety-nine specimens of Physopsis africana harbored Bilharzia 
cercariae. These are characterized by the absence of a pharynx and 
by a divided tail. One specimen obtained from the Durban brickfields 
on May 9 harbored cercariae with long undivided tails, as well as a 
sporocyst containing an eye-spotted form of furcocercous cercaria. 
This is the only specimen of the kind I have seen, and, in consideration 
of its resemblance to the Egyptian form, I have suggested for it the 
name Cercaria oculata. The eye-spots had a crescentic appearance and 
were situated nearer the oral end of the body, on either side of the 
divided gut. They could be readily seen through the thin walls of the 
sporocyst in which the cercariae were well developed. No pharynx 
was discernible in the body of the cercariae. There was a long, slender 
tail which was divided into two short, fin-like prongs. 


CAWSTON—CERCARIAE OF NATAL 133 


CERCARIA SECOBIANA 


A common cercaria from the Umsindusi pool, for which I have 
suggested the name Cercaria secobiana, occurred in about seventy 
specimens of Physopsis africana. It was narrower andslightly smaller 
than the eye-spotted form. This distome had a long, slender tail which 
was divided into two prongs. The prongs were as long as the tail 
itself. When the tail moved, the prongs became bent to the form of a 
crescent, causing the cercaria to swim backwards —a form of locomo- 
tion which would seem to be common to furcocercous cercariae. The 
cercariae developed in a sporocyst which intersected the whole liver- 
substance of an infected specimen. They were found only in Physop- 
sis africana from the Umsindusi River. At present, no light has been 
thrown upon the life-history of this cercaria, which has the appearance 
of an avian trematode. 


SCHISTOSOME CERCARIAE 


Cercariae which answered to the description of the Schistosome 
group were found in sporocysts from the liver-substance of twenty- 
three specimens of Physopsis africana (15 per cent) from the brick- 
fields at Durban. They were present in a lesser proportion of speci- 
mens of this same mollusc collected from the Umsindusi River. Bil- 
harzia disease is common amongst bathers in both these places. Except 
for the absence of eye-spots, the cercariae were identical with the eye- 
spotted form. The long, slender tail was divided into two short, fin- 
like prongs. There was no pharynx to be seen. In the Medical Journal 
of South Africa for April, 1916, Dr. J. G. Becker reported that these 
distome cercariae occurred in Physopsis africana from a pool at 
Nijstroom in the Transvaal. I have seen the microscopic preparations 
he has made of them. He injected some hypodermically into a guinea- 
pig and, as I announced at a meeting of the Witwatersrand Branch of 
the British Medical Association two months later, succeeded in pro- 
ducing three adult male Bilharzia worms in the portal system. This 
confirmed the opinion that these cercariae, which have only been found 
in areas known to be infected with Bilharzia disease, are in reality the 
larval form of the Bilharzia parasite of man. 

On April 28, I added some water containing miracida serena 
from the urine of a patient suffering from Bilharziasis to a vessel of 
water containing specimens of Physopsis africana from the Umsindusi 
River. At the end of a fortnight a small sporocyst containing unde- 
fined cercariae was seen throwing out branches throughout the liver- 
substance of one specimen, giving it a yellowish-white appearance. In 
another specimen a similar young sporocyst occurred; in this could be 
seen undeveloped cercariae with bifid tails. By the end of three weeks 


134 THE JOURNAL OF PARASITOLOGY 


fourteen out of thirty-one specimens, or 45 per cent, harbored Bil- 
harzia cercariae, while only 15 per cent of specimens obtained direct 
from the river were found to. be infected at that period of the year. 
In another series of experiments, the addition of miracidia to the water 
in which specimens of Physopsis africana were kept, apparently 
increased the number of infected forms from 22 to 37 per cent, and 
from 23 to 27 per cent. Similar experiments with specimens of 
Planorbis pfeifferi and Limnaea natalensis proved entirely negative. 


SNAILS HARBORING “TADPOLE” CERCARIAE, 1916 


Date Source Species No. Infected Percentage 
ADIT, cocoate er Umsindusi.....<...: Limnaea natalensis..... 2 out of 88 
(Pietermaritzburg)..| Limnaea natalensis.....| 0 out of 30 1.6 
(Pietermaritzburg)..| Limnaea natalensis.....| OQ out of 6 
(Pietermaritzburg)..| Physopsis africana..... 12 out of 197 
| (Pietermaritzburg)..| Physopsis africana..... 4 out of 200 4 
(Pietermaritzburg)..| Physopsis africana..... | Ooutof 6 
| ‘Toll Gate............ Limnaea natalensis...... 7 out of 47 
\GDaEban) i pacsc.nssocns Limnaea nDatalensis..... | loutof 12 13.75 
| (Durban)... 0226s Limnaea natalensis...... 3 out of 21 
(Durban) s....¢..- 2. Physopsis africana..... | ‘Lo out- of “7 
HGDIIT DAN) coe cee oetecice Physopsis africana..... 7 out of 85 5 
GDuTDAN) Se eccccieewses Physopsis africana..... 0 out of 13 
(DELDAaD) serccs cers Physopsis africana..... 4 out of 131 
(Prarban) fs ccc gsc =. Planorbis pfeifferi...... Tout of 24 
(Dieban) csc ececcne: Planorbis pfeifferi...... 49 out of 163 30 
.| (Durban).. ..| Planorbis pfeifferi...... Tout of 20 
(Gd iridef 00) Sea Planorbis pfeifferi...... 3 out of 13 
(Durban) <e.sce ces. sc Isadora tropica......... 0 out of 56 0 
Umegeni (Durban)....| Planorbis pfeifferi...... 5 out of 15 13.33 
BOshod, St.cseese-es Physopsis africana..... 0 out of 20 0 
(Pietermaritzburg)..| Physopsis africana..... 0 outof 6 
(Pietermaritzburg)..| Ancylus.......... Saccidcets|| SOLON (OL 220 0 
(Pietermaritzburg)..| Isidora forskali......... O out of 2 | 0 
(Pietermaritzburg). .| Isidora forskali......... | Oout of 1 | 
PHYSOPSIS HARBORING BILHARZIA CERCARIAE, 1916 
Month Source Number Percentage 
ADTs. Sep ncceasncemeitete OMG G RTGS 5 oan ow clnie occ tiacctet res > ete | Lloutof 7 
MB c.\'0 sven aneuienenmeae KOT EADISBARY) vies cote a’ fara oiola, genre Olas eters scion 13 out of 85 10 
SUNG. ce vocessstoenehe sees CUVGTAT ATI) S. cass Svein anemeaeans neve | 2 out of 13 
CU SAR Ane pricdascrr ac (LD) OF Ope ae Se A eee 8 out of 131 
ADIL, S.-i re cecaee eines DUMAING BL: « oiacciare arasien eee a 80 out of 197 |) 
MOY) .'.0.55. Serie eee (Plotermaritzbure)\..... S22cesee5 case | 388 out of 170 || 18.6 
DUNC 63 sy cle dace eee | (Pietermaritebure)... 26 52cchcenaces 7 out of 30 | 
iti k Caeser! (Pietermaritzburg): .... Cee. ce ceces Oout of 6 
DUNG. c's as oes nd eee ee ote BORO Wt. lOO... se cae e eae ce 0 out of 20 i 0 
Vitae cle aioe ase aoe (PletermaritZDure)’<:<:-..tebosee< sie 0 out of 6 


CAWSTON—CERCARIAE OF NATAL 135 


With the exception of the Schistosome cercariae, we are at present 
entirely ignorant of the life-history of the various South African cer- 
cariae. Some of the “tadpole” forms may give rise to the flukes which 
occur in sheep and cattle in certain parts of the Transvaal, Natal, and 
Griqualand East. Others may produce the flukes which I am told are 
common in the lungs of frogs from some of the pools and rivers of 
Natal; but, as stated in a letter from Sir Arnold Theiler of the Agri- 
cultural Department, “Nobody has yet undertaken to work out the 
life-history of these flukes in South Africa, and the only reference is 
that given by Doctor Gilchrist in his book on South African Zoology.” 

The importance of this study is emphasized by our need of a drug 
to destroy the adult forms of cercariae in the human host. Perhaps a 
drug which would destroy the liver-fluke in sheep would be equally 
efficacious in destroying the Schistosome parasite of man. 


NOTE ON A SPECIES OF NOSEMA INFECTING 
; ATTACUS CYNTHIA DRURY 


SHIGETANE ISHIWATA 
The Imperial Sericultural Experiment Station, Tokyo, Japan 


While working on dead larvae of Attacus cynthia Drury I met with 
many individuals which were infected by a species of Nosema with 
spores characterized by shape and refraction of light sufficiently dif- 
ferent from those of the silk-worm parasite, Nosema bombycis Nageli, 
to distinguish it from that species, although more closely allied to it 
than to any other species of the genus. 

The following. note is meant to give a short ieee of the 
structure of the spores thus far observed, the details of the same as 
well as the life-history of the parasite being reserved for further 
investigations. 

The spores taken directly from the body of the worm were fixed in 
sublimate-alcohol and stained with Giemsa’s solution. Heidenhain’s 
and Delafield’s hematoxylin were also used. For the protrusion of the 
polar filament I employed the method which Kudo (1913) tried with 
success for the spores of Nosema bombycis Nageli. 

The spores (Figs. 1 and 2) are ovoid, tapering toward each end. 
The refraction of light under the microscope is not so sharp as those 
of other species. On account of the taper they look long and narrow, 
like the spores of the species infecting Anthaerae pernyi Guér. and 
A. yamamai Guér., but measurements show that this is not the case, 
the length and the breadth of the spores of the present species being 
3 to 3.54 and 2p, respectively, like those of Nosema bombycis, the 
measurements of which given by Kudo (1916), were 3 to 4» in length 
and 1.5 to 2 in breadth, by Stempell (1909) 4 in length and 2, in 
breadth, and by Omori (1912) 2 to 4» in length and 1 to 2 in breadth. 

The spore is covered by a thick membrane of a transparent and 
homogeneous substance like that of N. bombycis, but as mentioned 
above, the refraction of light is not so sharp as in that species. The 
inner membrane, observed for the first time by Kudo (1916), also 
exists in the present species and can be pressed out easily. The outer 
membrane, however, appears to be rather brittle, as it is liable to be 
crushed into two or more pieces during the process of pressing. With 
an India ink preparation (Burri’s method) the protoplasm stains 
slightly black, as does that of the silk- worm Nosema (Figs. 1 and 2). 

The polar filament can be easily extruded from the end of the spore 
by the method used by Kudo (1913) for Nosema bombycis. It is 
somewhat shorter and thicker than that of the latter, being about 30 
to 35p in length. The filament ends always in a round knob of special 


ISHIWATA—NOTE ON A SPECIES OF NOSEMA 137 


form (Figs. 3, 4, 5 and 6), which is most probably of a sticky nature. 
In some specimens of N. bombycis, the polar filament has a round end, 
but not in all, and this again is not so well pronounced as in the present 
species. Moreover, the filament is not coiled transversely within the 
spore as in that of N. bombycis or of N. anomalum Monz., as is 
assumed to be the case by Stempell in these species. Careful observa- 
tion of the spore under a high power shows a clear line running longi- 
tudinally within the body, which in some appears to be placed in 
parallels (Fig. 7). These lines undoubtedly represent the filament 
coiled up within the polar capsule; and when the filament begins to 
uncoil and a part of it protrudes from the body of the spore, concentric 


Ss 
_@d om 


4 5 = > 
oe ee 


(All the figures are drawn with the help of an Abbé-Zeiss camera.) 


Ss 


Figs. 1 and 2.—Spores with protoplasm stained slightly black. India ink 
preparation. In Figure 1 narrow and Figure 2 broad. > 1545. 

Figs. 3 and 4—Spores with extruded polar filaments. The end of the fila- 
ment has a round point. 1545. (Figure 3 shows some crushed pieces of outer 
membrane which are deeply stained.) 

Figs. 5, 6, and 7.—Spores containing coiled filament. In Figures 5 and 6 
filament partially extruded. > 1545. 

Fig. 8—Polar filament extruded from side of spore. 1545. 


lines can often be seen within the latter (Fig. 6). Sometimes the 
filament extruded from the side of the spore takes the form of a ring, 
which is most probably caused by the sticking of the round end of the 
spore at the base of the filament (Fig. 8). 

Finally, I wish to express my hearty thanks to Prof. Dr. C. Ishi- 
kawa for his kind advice in the preparation of this paper. 


Papers CITED 


Kudo, R. 1913. Eine neue Methode die Sporen von Nosema bombycis 
Nageli mit ihren ausgeschnellten Polfaden dauerhaft zu praparieren und deren 
Lange genauer zu bestimmen. Zool. Anz., 41: 368-371. 

1916. Contribution to the Study of Parasitic Protozoa, I. On the Structure 
and Life-History of Nosema bombycis Nageli. Bull. Imp. Sericult. Exp. Sta., 
Japan, 1: 31-51. 

Omori, J. 1912. Zur Kenntnis des Pébrine-Erregers Nosema bombycis 
Nageli. Arb. Kais. Gesundh. Amt., 40: 108-122, 1 pl. 

1916. Nihon Sanbyo Ron. 

Stempell, W. 1904. Ueber Nosema anomalum Monz. Arch. Protistenk., 
4: 1-42, 3 pls. 

1909. Ueber Nosema bombycis Nageli. Arch. Protist., 16: 281-358, 7 pl. 


NOTES ON POROCEPHALUS GLOBICEPHALUS 
THESLE T. Jop anD A. R. Cooper 


Mary L. Hett of the London Zoological Society described and 
named Porocephalus globicephalus from a single mature female speci- 
men, procured from the lung of an American specimen of “moccasin,” 
Tropidonotus fasciatus (Linn.). In the Proceedings of the Zoological 
Society of London, 1915, pages 115-121, she gives the following char- 
acters: length, 50 mm.; annulations, about 50; hooks, simple and 
sharply curved; mouth, pear-shaped with pointed anterior end; head, 
globular; well marked neck; anus transverse slit on terminal segment. 

The above description, which is’ necessarily meager, is the only 
reference the writers can find to this species. In view of this con- 
dition the following data are herewith reported: 

A large black snake, Basscanion constrictor (Linn.), was received 
at the State University of Iowa in the fall of 1916 from Garrison-on- 
Hudson, New York. When the specimen was killed five males and 
five females of P. globicephalus Hett were found in the respiratory 
tract. Three males and three females were taken from the lung and 
two males and two females from the dorsal body wall of the air sac. 

The females were found with the head only embedded in the lung 
tissue, or (those in the air sac) in the muscuiature of the body, where 
a copious hemorrhage had been formed. The rest of the body of the 
parasites was free from attachments, hanging limply in the lumen of 
the lung or air sac. The heads of the males were not embedded in the 
tissues of the host, but only superficially attached to the walls of the 
lung or air sac by the hooks. 

The females vary from 82 to 96 mm. in length, being somewhat 
larger than the specimen described by Hett, while the males were from 
14 to 30 mm. long. The color of the female is lemon yellow, the body 
wall being transparent, thus permitting easy observation of the mass of 
embryos and the movement of the intestine within. The male is pale 
cream in color and the body wall is opaque. 

The head is globose dorsally; ventrally it is slightly concave with 
four sharply curved hooks at the anterior edge of the concavity, two 
on either side of the pear-shaped mouth. The neck is markedly con- 
stricted ; the body subcylindrical, slightly tapering to the posterior end 
which is blunt ; the digestive tract is seen from the dorsal side; laterally 
an opaque band runs the full length of the body (this becomes trans- 
parent in specimens preserved in alcohol, while the rest of the body 
becomes opaque). There are about 50 annulations, 48 to 52 having 
been counted. The digestive tract, which is gorged with blood, is 
readily seen in the living specimen, and may be traced in preserved 
ones. 


BOOK REVIEWS 


THE ANIMAL Parasites oF MAN. H. B. Fantham, J. W. W. Stephens, F. V. 
Theobald. New York: William Wood and Company, 1916. xxxii +901 
pages. 423 figures. $12.00. 

As the title page indicates, the work is adapted from the fourth German 


edition of Braun’s well known treatise. It appeared simultaneously in London 
and New York just after the publication of the fifth German edition of Braun 
which was reviewed in the JourNAL for June, 1916. A comparison of the views 
expressed in this work with those in the fifth edition of Braun is especially 
interesting as it shows the conclusions reached entirely independently by two 
groups of writers in the same field. It is not surprising that each book treats 
with greater fulness the work done by investigators of the same nation as the 
authors, and passes over more briefly the results achieved by workers in 
foreign nations. 

The plan of the work is ideal: the section on Protozoa was entrusted to 
H. B. Fantham, that on worms to J. W. W. Stephens, and that on the 
Arthropoda to F. V. Theobald. At the present day the work accomplished 
in each of these fields is so great and the questions under discussion so involved 
that no one man can cover them all with equal proficiency. Under the plan 
adopted here each field is assigned to an investigator who is qualified to 
write with ultimate authority on the problems in that field, and it would have 
been difficult to select three men anywhere who would measure up to the 
ideal better than those chosen. 

On the other hand, the time was not particularly propitious for a great work. 
Other things are in the air that make insistent demands on the attention of 
all men. There is no leisure for reflection, and concentration on a scientific 
problem must be well nigh impossible for a man working anywhere in Europe. 
In truth the book itself shows some evidence of present conditions in the world. 
It contains a wealth of information on little known topics. It has been brought 
thoroly up-to-date, even to the extent of two appendices including important 
material of later date than the general text, and further in having very recent 
items incorporated on slips bound in between the finished pages at the last 
moment. This makes the work appear confused, and even in the text there 
are places where the same impression is given the reader. It seems as if 
the authors had been working under pressure and the finish one expects in a 
masterful production had been marred. The volume of scientific material pre- 
sented to the worker is large and in every way equal to one’s expectations, 
but it is not equally well assimilated at all points. The practice of adding 
paragraphs here and there incorporates new material at the expense of fluency 
and the text is not always easily read and understood. 

The bibliography is very extensive, covering some pages in fine type, but 
it lacks all recent items, being in fact but a reprint of the lists in the 1908 
edition of Braun. Some references to the literature of the recent items in 
the text are given in footnotes; but in too many places the new facts are 
recorded without exact credit, or sometimes without even the name of the 
author, and the student is left to hunt for himself the precise source of the 
information. This is least noticeable in the section on Protozoa where footnote 
references are particularly abundant. It is curious to note that even with such 
_a large space devoted to bibliography one cannot find references to important 
recent papers by Leiper and other English workers; the authors have treated 
everyone impartially as not only other references are wanting but even some 
important papers of Stephens himself are not listed. 

The illustrations are numerous and well distributed. They include fewer 
relics of the past than one usually finds in so comprehensive a work. Most 


140 ~ THE JOURNAL OF PARASITOLOGY 


of the figures are well done and satisfactorily reproduced. One can not help 
feeling a little disappointed, however, that some have crept in which are new 
and yet unfortunately inferior. The diagrammatic representations of -the 
Echinococcus cysts on pages 352 and 353 are not well drawn and their repro- 
duction on so large a scale is still more open to criticism. A reduction to 
one-half or one-third the present size would have made their sketchiness less 
conspicuous without the loss of any important details. In the opinion of the 
reviewer English and American works are distinctly inferior to continental 
publications in the character of their illustrations, and the present volume is 
undoubtedly less deserving of criticism in this respect than most of our works. 
Despite its evident minor imperfections this treatise is a valuable and usable 
work. No one can question that the splendid volume is easily the largest and 
most complete work on the subject which has appeared in the English language. 
The work of the printer has been well done and deserves especial commendation. 
Both paper and type are such as contribute to ease of reading and one lays 
the volume aside with the conviction that its authors and publishers alike 
deserve the thanks of scientific workers for the results they have achieved. 


JapANesE MepicaL LITERATURE, a review of current periodicals the initial num- 
ber of which was noted in the Journal (3:42), has completed its first volume, 
July to December, 1916. The General Index is well prepared and will be a 
real convenience even though complicated by the fact that it is paged after the 
China Medical Journal from which the reviews are reprinted, and not according to 
the reprints themselves. References to animal parasites are numerous -and 
important. The value of the Japanese literature and its great inaccessibility 
make such reviews of unusual service and American investigators are deeply 
indebted to Dr. Mills and his colleagues for their work. 

The first number of the second volume has also come to hand, and scientific 
workers gladly look forward to the continuance of this very valuable serial 
which is without any competition in this difficult and important field. 


NOTES 


“RECHINORHYNCHUS MONILIFORMIS” IN NORTH AMERICA * 


Among the Acanthocephala which are exclusively parasitic and highly spe- 
cialized for the parasitic, habit, only three species have been reported for the 
human host and even these are rare or doubtful. Gigantorhynchus hirudinaceus 
(= G. gigas) is said to occur in man in southern Russia but the statement is 
unconfirmed. Lamb! found in the intestine of a boy a single parasite to which 
he gave the name of Echinorhynchus hominis. 

The third species, originally named Echinorhynchus moniliformis, has com- 
manded especial attention by virtue of its relation to man. Grassi and 
Calandruccio found it in Sicily in the small intestine of field mice, rats, and 
marmots. They detected the intermediate host in Blaps mucronata and in 
some cases found as many as 100 larvae encysted in a single cockroach. They 
fed such larvae to a white rat and Calandruccio swallowed some at the same 
time. These developed well in both hosts. The authors identified eggs appar- 
ently of the same species in the feces of a young peasant but were unable 
to carry out a cure and confirm the diagnosis. 

Through the courtesy of Mr. G. E. Clark some material has been placed 
in my hands which belongs to a larger species of closely related type. These 
worms were taken from a squirrel in Illinois. They furnish the first record 
of this type for the North American continent. As noted above the European 
species has been grown experimentally in the human host and this species is 
likely to show the same power if the mature larvae are introduced by any 
chance into the human intestine. 

After extended study it may be said that the two species are both sufti- 
ciently characteristic in their resemblances to each other and in their differ- 
ences from other known forms to constitute a new genus to which the name 
of Hormorhynchus may be given. H. moniliformis (Bremser 1819) is desig- 
nated as type and attention is called to the fact that Liithe believes there are 
several species in Europe all included under the one name. The American 
species is designated as H. clarki. Specimens measure 100 to 130 mm. in length. 
The proboscis is very small, being only 0.255 mm. long by 0.12 mm. broad. The 
first ring is 5 mm. from the anterior tip. The rings begin faintly but distinctly ; 
they are about 1 mm. long and little wider than the body. They increase rapidly 
up to 2 mm. in length and at the point of greatest length the individual rings 
are so swollen as to become markedly wider than the body. From this region 
they taper out very gradually. The last 15 mm. of the body shows no trace of 
rings and for the same distance anterior to it the rings are very faint. A 
full description of the species will be published elsewhere. 

Henry B. Warp 


DIPTERA IN THE HUMAN INTESTINE 


On Sept. 30, 1916, Dr. W. C. King of Helena, Ark., sent some intestinal 
parasitic worms which he reported as coming from an adult woman, to the 
State Hygienic Laboratory connected with the Medical Department of the 
University of Arkansas for identification. Dr. A. C. Shipp, in charge of 
the Hygienic Laboratory, consulted with me about these worms. They were 
plainly annular with about thirteen segments. The shape was very nearly 
pyramidal with some seven or more papillae at the blunt posterior end. 


* Contributions from the Zoological Laboratory of the University of IIli- 
nois, No. 91. 


142 THE JOURNAL OF PARASITOLOGY 


These worms looked so much like Dipterous larvae that I suggested that 
we place them on nutrient agar for a few days to see if they would pupate. 
Pupation took place in about three days and after about five days more 
there was hatched a medium-sized black fly. Later others were hatched. Some 
of these were sent to Prof. James S. Hine of the Entomology Department of 
Ohio State University, who identified them as Sarcophaga assidua Walker. 

Our attention has been called to two or three other cases of similar occur- 
rence of what seemed to be Dipterous larvae in the stools of persons this 
past fall and summer. In one case the sputum gathered in a sterile bottle 
under a physician’s guidance showed what seemed on examination to be 
young Dipterous larvae. Unfortunately these larvae were killed in the over- 
heating of our incubator. 

How these larvae gain access to the intestine unless through ingestion of 
food in the stage of the freshly laid eggs on exposed cooked food or in 
uncooked food, is a question. Whether they passed the gastric digestion in 
the stage of egg or larva is problematical. CHARLES BROOKOVER 

Department of Anatomy, University of Arkansas. 


Dr. E. Gonzalez reported to the Congress of Medicine at Maracaibo the 
occurrence of Leishmania brasiliensis Vianna in a servant woman brought to 
the clinic at San Fernando de Apure. The preparations were examined by 
members of the Yellow Fever Commission of the Rockefeller Institute and the 
diagnosis confirmed. This is the first case of cutaneous Leishmaniasis from 
Venezuela. 


Parasitologists will be interested in the circular of the Merchants Associa- 
tion of New York on the Dangerous House Fly. It is important to urge on 
every community the necessity of early action this year to eradicate this 
dangerous agency in the transmission of disease. Incidentally also laboratory 
teachers may be glad to know that in the flies which appear early in the season 
a flagellate (Crithidia sp.?) occurs abundantly in the gut. Material can be 
secured readily from this host that is well adapted to class study.or demon- 
stration of this group. 


The Journal of Parasitology 


Volume 3 JUNE, 1917 Number 4 


ENDAMOEBA BUCCALIS 


I. ITS MULTIPLICATION AND PERIODICITY 


NADINE NoOwWLIN 
University of Kansas 


Work on these much-discussed parasites was begun with a two-fold 
object: (1) to find the cause of their periodic disappearance from the 
human mouth, this phenomenon pointing to a solution of the life- 
history which has not been solved; (2) to determine by tonsil smears 
and sections whether this endamoeba is ever intracellular. 


MATERIAL 


The specimens used in this study were collected entirely from one 
host and frofh one point of infection, an upper premolar tooth which 
was the single focus of infection for a long while. Daily record was 
kept of the occurrence of these parasites for a period of over five 
‘months, from May 1 to July 29, 1915, and from October 10 to Decem- 
ber 12, 1916, as well as of health conditions of the host. 

Moreover, a study of the parasite was made at various times of 
day and night with a view to determining whether behavior varies 
with these conditions. 

My attention was first called to the suspected periodic disappear- 
ance of Endamoeba buccalis by one of my students, who was working 
on this parasite in the zoological laboratory of the University of Kansas 
during the winter of 1914. While the supply of material was usually 
generous, at times she reported it too scarce for study. Since then 
other protozoology students have reported finding it at times and 
being unable to do so at others. This did not seem to be wholly due to 
mouth cleaning, and suggested to me the possibility of migrations of 
the parasites into the gums or periosteum at the so-called “scarce times.” 

While I set out with this as a pure hypothesis, my observations 
during five months have convinced me that E. buccalis appears and 
disappears, tho with not so much regularity as I had at first thought ; 
my journal shows nine scarce times in five months. 


144 THE JOURNAL OF PARASITOLOGY 


Since I have no definite proof for the ideas I am about to set forth 
in explanation of these migrations, I offer them only as strong circum- 
stantial evidence, with the hope that the suggestion may serve as a 
good working clue. 


METHODS OF STUDY 


For diagnostic purposes the fresh smear containing the living 
material is best. A small amount of scrapings from the point where 
the tooth joins the gum was spread on a cover-slip slightly heated. 
This was then placed upon a slide containing a small drop of normal 
salt solution. If a prolonged study is desired, the slip can be ringed 
with vaselin before being placed on the slide. I found this method 
much better than smearing the slide and then covering with normal 
salt and cover-slip, or than mixing the scrapings freely with the salt 
solution. The natural environment is most nearly reproduced by the 
hanging drop method, and it may be due to this that I was able to 
observe behavior which, so far as I know, has not been reported for 
this amoeba. 

Among intra vitam stains, neutral red differentiates the food 
vacuoles very quickly, but has the disadvantage of staining the whole 
parasite intensely in a short time. It brings out few points which the 
unstained, living specimen does not show. After a little experience, 
there is no danger of confusing the unstained endamoeba on a slide 
with the leukocytes. The endamoebae have a greenish refractive look 
which differentiates them even under the low power, where they appear 
smaller than a pin head. Even motion is not necessary. 

For nuclear studies, further investigation of food bodies, structure 
of cytoplasm, etc., material was preserved, stained according to various 
methods of protozoa technic and compared carefully for results. Need- 
less to say, the wet film method was adhered to thruout except in one 
process, and that at the very end of the Giemsa stain, just before 
mounting in balsam. Comparasion of a slide thus treated with one 
run into xylol and not allowed to dry, showed no ill effects, and the 
stain was better without the xylol treatment. 

The three methods generally employed were: 

(1) The short Giemsa. Bring the wet smear into half and half 
methyl and ether for 5 minutes. Transfer to a solution of Giemsa 
made by adding 1 cc. of stock solution to 15 cc. distilled H,O for § 
to 10 minutes. Wash with distilled water until pink appears. Barely 
dry in air and mount in balsam. . 

This gives a beautiful differential stain for the amoebae, coloring 
the cytoplasm pale blue, their food vacuoles wine, and the nucleus vivid 
red. This stain brings out a halo of chromidia around the nucleus 
which other stains do not. Giemsa also differentiates leukocytes clearly 


NOWLIN—ENDAMOEBA BUCCALIS 145 


from even the smallest endamoeba because leukocyte cytoplasm stains 
pink and their nuclei deep lilac. Epithelial cells take a faint pink in 
the cytoplasm, and a deep pink nuclear stain. 

(2) Fixation with the methyl-ether mixture compares favorably 
with the picro-mercuric-formalin method, which is recognized as one 
of the most satisfactory fixations for protozoa. This latter fluid used 
hot and allowed to stand on the smear until it has evaporated almost 
completely, then washed in 70 per cent alcohol and stained by Dobell’s 
quick hematein method, gives a very clear image of the nucleus, if 
properly differentiated. 

(3) A very similar effect is obtained by hot Schaudinn’s fluid 
(80 parts Mg Cl, plus 20 parts absolute alcohol) followed by Heiden- 
hain’s iron-hematoxylin used unripened. 

Mallory’s stain recommended by Craig (1911) for sections was 
used on tonsil sections suspected of containing endamoebae, as well 
as on fresh smears of Endamoeba buccalis without satisfactory results. 


REPRODUCTION 


Most of the endamoebae found in smears contain nuclei in the 
resting stage; and this as pictured in most texts and as shown also in 
my own slides by the mercuro-picric-formalin fixation and either Dobelt 
or Heidenhain’s iron-hematoxylin stains, has a nuclear diameter 
scarcely one eighth the diameter of the amoeba, a well defined mem- 
brane, with its inner margin lined with chromatin granules, and a 
central body or nucleolus (Fig. a). A clear area surrounds the nucleus 
in fixed material, and since it has been impossible to study it in the 
living condition, it cannot be determined whether this area is due to 
shrinkage or is what shows as a pink halo with the Giemsa stain 
(Figs. e and 7). 

Material stained with Giemsa shows outside of this so-called nuclear 
membrane, and in about the region of the clear area mentioned above, 
a halo of rose-colored granules, which I interpret as chromidia. Every 
nucleus stained thus shows this, and I conclude that it is present even 
in the very earliest resting stage of all Endamoeba buccalis, but is net 
brought out by certain stains. 

Craig (1916) believes E. buccalis has a primitive type of mitosis. 
My observations point toward a complex one. During the early stages 
of mitosis the nucleus enlarges and the karyosome disappears. . Chro- 
matin collects on the nuclear membrane, making an irregular border 
(Fig. e) and leaving a clear central vacuole. At times the chromatin 
of this phase is so organized that it gives the appearance of a spireme, 
except that the vacuole at the center is always present. A halo of 
chromidia surrounds the dense chromatin and no doubt contributes to 


146 THE JOURNAL OF PARASITOLOGY 


its formation, since it grows paler as the chromatin condenses. The 
chromatin condenses into four very distinct bodies and the nucleus 
remains in this condition longer than in others, judging by the large 
number found on a slide in the prophase of mitosis. 

Whether these four bodies can be compared with chromosomes of 
higher animals, I do not know, because as soon as the spindle forms 
they mass and become indistinguishable. Yet they indicate very definite 
organization during at least part of mitosis. Figure g is the typical 
metaphase, Figure 4 the anaphase, and Figure i the telophase. In 
comparison with early stages of the nucleus these later spindle stages 
seem very small, the difference being due to the chromidia outside what 
is usually designated as the nuclear membrane. I doubt now whether 


ke 


Endamoeba buccalis. For details see text. 


a nucleus as represented in Figure a is the entire chromatin mass, since 
I have found chromidia in those of about the same phase stained with 
Giemsa (Fig. b). 

The spindles are distinct and give the appearance of having a cen- 
trosome at the poles. The size of the nucleus varies considerably, the 
larger ones being lodged usually in the larger amoebae; but there are 
frequent exceptions to this (Fig. /). 

I have occasionally found two equal-sized endamoebae lying in close 
contact and suggesting binary fission. A close study of these, however, 
has never revealed a dividing nucleus, but two well formed and widely 
separated ones. The cytoplasm was in every case completely separated. 
Very frequently two living endamoebae are seen gliding over each 
other and becoming so nearly fused that at times the most careful 


NOWLIN—ENDAMOEBA BUCCALIS 147 


observations cannot distinguish two separate animals. I have suspected 
that there may be an interchange of materials between two such indi- 
viduals, but I have never been able to confirm Craig (1916), who says 
he saw streaming of cytoplasm from one to the other. 

The incessant gliding of the amoebae over each other gives this 
appearance. This may be conjugation. I have never seen buccalis in 
the actual process of division. 

I have never seen buds form and become separated from the parent 
cell and then develop, tho endamoebae giving the appearance of bud- 
ding are frequent. At such times the cell resembles the pearl-stage of 
gregarines, but long enough observation has usually seen them with- 
drawn. It is usually an adverse condition, such as drying-up or low 
temperature, which causes this appearance. When proper conditions 
are restored, normality of form may be resumed. 

I have seen about half a dozen stained specimens which I interpret 
as multiple fission. This is a small number out of the hundreds ct 
specimens passed in review, and yet multiple fission is probably a rare 
process, which does not take place in the mouth cavity. The suspected 
forms have no protective walls, have usually been found in close con- 
tact with leukocytes or epithelial cells, and are somewhat irregular in 
outline (Fig. k). 

There is no limited number of merozoites as eight or four in the 
E. coli and E. histolytica cysts, but the number may vary from eight or 
nine to more than a dozen. Those I have seen do not seem compara- 
ble to the reproductive cysts of E. coli and E. histolytica, but suggest 
rather the merozoite formation in Plasmodium, and probably serve 
merely to spread the infection in the host. 


Endamoeba buccalis follows the course of all protozoa in encysting. 
It first becomes spherical and inactive and begins to diminish in size. 
I induced a kind of encystment once by leaving EF. buccalis sealed on a 
slide for six hours with the temperature gradually going down. These 
did not in that time change much in size, but the food vacuoles paled 
and seemed to dissolve in the cytoplasm. 

Normally, encysted forms are from one-half to two-thirds the size 
of the active trophozoite ; they usually show some faint, rounded inclu- 
sions, probably the remains of food vacuoles, and a clear wall slightly 
spaced from the animal protoplasm proper (Fig. 1). Encystment 
seems to be for protection against adverse conditions rather than for 
multiplication, as is shown by the following observations. 


RELATION OF HOST AND PARASITE 


Encystment with this form is not as rare as Craig (1916) believes. 
One strain followed daily for months will show the encysted condition 
from time to time, and my records show that encystment in Endamoeba 


148 THE JOURNAL OF PARASITOLOGY 


buccalis is closely connected with the “scarce periods.” As stated pre- 
viously, daily examinations showed numerous active endamoeba in the 
scrapings from a tooth for ten and fourteen consecutive days. Then 
would intervene two to four days when few could be found, though 
the same region was carefully explored. Such forms as may be found 
at these scarce times are recorded as “sluggish,” “spherical,” “encysted.”’ 
Most of the active forms left were small. Very deep probings into 
the gums around the tooth sometimes procured a few larger ones, tho 
finally even these would fail. 

Often when the parasites became numerous again they were slug- 
gish and half encysted (Craig’s precystic stage) for a day; then they 
became active and flourished again as usual for two or more weeks. 
Once they did not disappear for four consecutive weeks, but at the end 
of that time they were gone completely. 

Now what is the explanation of this periodic appearance and dis- 
appearance? [I laid it at first to mouth-cleaning, until that was care- 
fully tested out. I then considered that changes in the host might 
account for it, and following out that clue I found by my journal that 
practically every period of scarcity was accompanied by a time of low 
vitality on the part of the host as manifested by some slight indispo- 
sition. Indigestion accompanied at least three of these disappearances. 
I concluded that physiological changes of the body, whether normal or 
abnormal to the host, change the chemical reaction of the body fluids, 
notably the saliva in this case. These changes may be too slight even 
to be analyzed, and yet they affect the very sensitive endamoeba living 
in that medium. As in the case of free-living amoebae, encystment 
follows when conditions change from the normal, so with buccalis, 
when the saliva changes in its chemical qualities, the parasite encysts 
either partially or wholly. It withdraws into the periosteum, however, 
for this purpose. 

Further evidence for this was furnished by extracting the infected 
tooth during a scarce period. The endamoebae had flourished for 
four weeks and then disappeared ; for two days none had been found. 
The tooth showed an ulcer at the root, and by aid of the binocular 
many lesions as described by Bass and Johns (1915) were seen on the 
periosteum. These lesions examined microscopically showed many 
leukocytes and few amoebae, but on the periosteum around them, and 
especially where it joined the tooth, amoebae, were extremely numer- 
ous. These seemed to be in very close contact with the tissue, tho some 
were removable by means of a fine scalpel. All of the endamoebae 
thus found were in partial or complete encystment. 

I think this withdrawal of Endamoeba buccalis to the gums to 
encyst explains why encystment has been so seldom reported. I believe 
further that the end of the reproductive phase, binary fission, occurs 


NOWLIN—ENDAMOEBA BUCCALIS 149 


normally either in the gum tissue or in close contact with it; for 
although spindles are frequently seen in smear preparations, actual 
division of the cytoplasm has never been seen. Since the gums of 
infected patients cannot be sectioned, I now have under way a study 
of the tonsils in the hope of throwing light on the intracellular phase 
of the life-history. An understanding of the periods of disappearance 
of E buccalis would be valuable in the treatment of pyorrhea if it be 
caused by this parasite. 


TONSIL STUDIES 


With a view to determining whether Endamoeba buccalis is ever a 
tissue dweller, I have studied the surface scraping of tonsils both before 
removal from the patient and after. In only one case did I find enda- 
moeba in the fresh smear, and their detection is certain if they occur. 
In no case have I been able to identify parasites in the paraffin sections. 
In view of the fact that Smith, Middleton, and Barrett (1914) found 
amoebae so plentifully on the tonsils examined, I have been surprised 
to find none in any of my examinations. If E. baccalts is a tonsil para- 
site, it seems very probable that it penetrates the tissue of so soft an 
organ and undergoes some interesting stages of development which I 
believe are constantly taking place in the gums, but which I have no 
means of demonstrating. 

As my work in this direction is scarcely begun, the negative results 
thus far are by no means conclusive or even discouraging. 


REFERENCES CITED 


Bass, C. C., and Johns, F. M. 1915. Pyorrhea Dentalis and Alveolaris. 
Jour. Am. Med. Assn., 64: 533-558. 

Craig, C. F. 1911. Parasitic Amoebae of Man. Philadelphia. 

1916. Observations upon the Endamoebae of the Mouth. 1. Endamoeba 
gingivalis (buccalis). Jour. Infec.-Dis., 18: 220-238. 

Smith, A. J., Middleton, W. S., and Barrett, M. T. 1914. The Tonsils as 
a Habitat of Oral Endamebas. Jour. Am. Med. Assn., 63: 1746-1749. 


ON THE SPOROZOON PARASITES OF THE FISHES OF 
WOODS HOLE AND VICINITY 


Il. ADDITIONAL OBSERVATIONS UPON MYXOBOLUS MUSCULI OF FUNDULUS 
AND A NEARLY RELATED SPECIES, M. PLEURONECTIDAE OF 
PSEUDOPLEURONECTES AMERICANUS 


C. W. Haun 


Reference to the multiplicative stages of this parasite was made 
in a former paper (Hahn, 1913). At that time the true parasitic 
nature of the trophoblasts of both the multiplicative and propagative 
stages was insufficiently established.. The relative virulence of the 
protozoon and the bacteria also needed further confirmation. Subse- 
quent studies leave no doubt as to either of these points. 

In almost every diseased integument, gill, or flesh wound which 
one examines, there will be found among the decadent tissues a few 
or many clear, white, even-contoured bodies which rarely take up any 
stain, no matter what treatment the tissues may be subjected to. The 
bodies are therefore in strong contrast with the surrounding tissues. 
If conditions are such that the parasites can be seen at all, the tissues 
must have taken up more or less of the stain. It was hoped that by 
using a variety of stains in different combinations with a wide range 
of fixatives, one might succeed in finding a treatment that would reveal 
the nucleus and perhaps other cytoplasmic contents of the parasites. 
No very encouraging results were obtained with the reagents that 
follow. 

After fixation with alcohol (Abs. 62 per cent), ether (32 per cent) 
and 40 per cent formaldehyd (6 per cent), I used Giemsa, toluidin 
blue, methylene blue, thionin, Bismarck brown, fuchsin, anilin blue, 
Bordeaux red, neutral red, dahlia violet, sudan III, indigo carmin, 
methyl violet, alizarin, rose anilin violet, carbol fuchsin, picro-nigrosin, 
safranin, and hematein combinations. With corrosive sublimate solu- 
tions in different solvents and after chromic, chromosmic, and many 
other common and some unusual fixatives, such as tannic, malic and 
formic acids, the following stains were employed: Ehrlich’s hema- 
toxylin, Mayer’s hematein, safranin, fuchsin, Heidenhain’s hema- 
toxylin, and brazilin. Both Mayer’s hematein and Heidenhain’s hema- 
toxylin give to the cytoplasm of the parasite a slight clouded effect 
which renders it visible throughout. Rarely a medium or large-sized 
trophoblast has a faint blue nucleus, and less frequently a small dense 
spherical nucleus. Brazilin has given promising results when used in 
connection with a 5 per cent aqueous chromic acid fixation. 


HAHN—SPOROZOON PARASITES 151 


The trophic stages of the multiplicative cycle are much more fre- 
quently encountered in all the tissues I have examined. They also 
occur in much larger numbers, especially the minute stages. Thousands 
of them are frequently distributed more or less equally throughout 
the myoplasm of certain areas of muscle fibers (Fig. 9). A few are 
interfibrillar. Such muscle fibers may or may not give evidence of 
hypertrophy. The size of the parasites in one and the same tissue may 
vary from 1.5 to 80 or 90p in diameter. A very good picture of them 
has already been published in Figure 12, Plate XX, of the paper 
mentioned above. The trophic stages of Chloromyxum clupeidae 
(Fig. 8) appear to be very nearly the same in appearance as those of 
M. musculi, 

In shape the multiplicative trophoplasts are circular or oval when 
small. Older ones have slight blunt extensions here and there over the 
surface. Occasionally a long pseudopod is encountered. Since these 
observations are made from fixed smears, it is probable that in life the 
display of activity on the part of the pseudopods would be very strik- 
ing, could it be seen. As yet I have observed no striking activity in 
numerous fresh tissues. In very large parasites the cytoplasm is 
finely granular. The smaller ones appear to be structureless. Tropho- 
plasts of moderate size frequently have a thin border of stainable mate- 
rial covering a part or all of the surface. This suggests an excretion 
or surface deposit, but is in reality what remains of the muscle nucleus 
which has been atrophied under the action of the parasite (Fig. 4). 
This can be demonstrated by the study of a large number of cases, 
when it will be found that there is a complete series of stages between 
the condition here described and normal nuclei. 

Multiplicative trophoblasts have been found in muscle epidermis, 
gill epithelium, and connective tissue. All of these tissues are attacked 
and undergo cellular degeneration. The nuclei and mucous cells 
usually remain in various stages of hypertrophy and constitute a very 
misleading series of artifacts. 

The staining reaction and the general appearance of the multiplica- 
tive trophoplasts are such as to suggest strongly that these bodies are 
some fatty or lipoid degeneration product. After many months of 
doubt, preceded by many more during which they were overlooked 
because it was assumed that the bodies in question were oil globules, 
it finally proved impossible to exclude them from the myxosporidian 
life-history. Authority may be found in the literature in support of 
both interpretations. It is an accepted fact (Adami, 1910) that with 
the hypertrophy of muscle, uniformly distributed fat bodies are to be 
expected. It has also been shown that the hypertrophy of the nucleus 
sets up changes in its immediate vicinity that result in lipoid substances. 


152 THE JOURNAL OF PARASITOLOGY 


The association of hypertrophied nuclei and Myxosporidia described 
above fits these specifications very well. On the other hand, small 
globular bodies within and between the muscle bundles were taken by 
Pfeiffer (1891: 106) to be germs of a myxosporidian. 

The evidence upon which I have based my decision is (1) the fail- 
ure of either osmic acid or sudan III to give a fat reaction, whereas 
oil globules on the same slides give a typical reaction. For the 
sudan III tests the tissues were fixed in aqueous formaldehyd solu- 
tion, treated with a low-grade alcoholic solution of sudan III, and 
preserved in glycerin. (2) The large trophoplasts show granular 
cytoplasm and a faint nucleus at times, when stained with Mayer’s 
hematein and Heidenhain’s hematoxylin. (3) The trophoplasts occur 
in graded sizes as if belonging to the same stage of growth. (4) Many 
trophoplasts have pseudopodial extensions that have a strong motile 
suggestion. (5) Many muscle fibers in an advanced stage of hyper- 
trophy are free from the bodies in question; they have migrated or 
operated in some other part. The products of degeneration would be 
expected to be uniformly distributed in all atrophied muscle fibers. 
(6) The sporoblasts of both M. musculi and Chloromyxum clupeidae 
have exactly the same oil-like appearance as the multiplicative tropho- 
plasts and reactions, but contain some characteristic body that belongs 
to the sporogenesis, such as the myxospore itself (Chloromyxum) or 
the sporoblast nuclei. (7) When one compares the trophic stages of 
the multiplicative cycle with the propagative cycle of the Myxobolus 
or both with similar stages of the Chloromyxum, four kinds of bodies 
may be recognized. If the structures in question are artifacts, this 
distinction into two classes would not conform exactly to the condi- 
tions required by the protozoon life cycle as to equality of development 
of all individuals present. This is exactly what is found in regard to 
both of the genera here described. Either all the parasites are young 
trophoblasts of the multiplicative cycle, or all are in some phase of the 
propagative cycle. (8) Many of my preparations have been treated 
with ether and absolute alcohol. Oils are extracted by this treatment. 
Yet most of the structures in question show some evidence of a solid 
content, whereas casts of fat bodies, when encountered, are clear and 
structureless. 

Many observers have found in fresh tissues small motile, structure- 
less bodies, and also cells with nuclei, which they have assumed were 
parasites. I have examined fresh infected tissues of both the herring 
and Fundulus. While able to recognize the trophoplasts and sporo- 
blasts, it has never been possible to be certain that the suspected 
objects were parasites until they were either connected by stages to 
sporocysts containing myxospores or until they had been verified in 
fixed and stained preparations. Pathological tissues frequently con- 


HAHN—SPOROZOON PARASITES 153 


tain artifacts resulting from the products of degeneration (Hahn, 
1913: 197) which are very misleading. There are also numerous 
tissue cells and ameboid cells with well-developed pseudopods in 
atrophied tissue, especially in the epidermis of such fish as the flounder. 
Under these circumstances, one is inclined to place little confidence 
in observations based upon fresh tissue alone. It is probable that the 
observations of Pfeiffer (1893 and 1891), Theélohan (1893), and 
Megnin on the trophic stages of M. pfeifferi were correct, but one 
must always feel doubtful about the reliability of one’s interpretations 
when good and sufficient reasons for considering any fresh cell as a 
parasite are not given. 

The multiplicative trophoplast continues to grow in size until it is 
over 50 in diameter. Although not observed alive, the shapes and 
general appearance lead to the conclusion that they are motile. They 
usually occur singly in comparatively uninfected portions of the tissue. 
In shape they vary from a long gregarine-like structure with very 
finely granular endoplasm and a shallow clear cytoplasm, to a smooth 
oval or circular mass when seen in profile. These large individuals 
may reach a diameter of 50y (Fig. 14). Associated with them in the 
same tissues one rarely finds schizonts containing minute spores. The 
schizonts range in size from 40 to 55y. These bodies are embedded in 
the muscle fiber in a cavity which they completely fill, giving precisely 
the same appearance as the large immature schizonts (Fig. 12). The 
multiplicative spores within are about 1.54 in diameter. In the few 
cases which I have examined, they have not taken up the stain, but 
are visible owing to the presence of a residual material which retains 
a moderately intense stain. Free multiplicative spores are common, 
and like all multiplicative stages, they are also characterized by the 
non-staining quality. Occasional schizonts containing spores are 
encountered in fresh tissues. They have also been seen and distin- 
guished from propagative stages in sections. As yet none of the latter 
were so large as those here figured. The scarcity of sporulating 
schizonts is no doubt to be attributed to the rapidity of the dissemina- 
tion of the spores under the muscular activity. As previously noted, 
the schizonts have already migrated into fresh tissues by the time they 
have reached any considerable size. 

One might suspect that the bodies produced by the so-called 
schizogony and figured here are bacteria. Very similar colonies of 
bacilli have been described elsewhere (Hahn, 1913). These bacilli 
do not occur in muscle which is essentially normal, and they are not 
accompanied by interstitial material when isolated and embedded in 
the myoplasm. The individual here figured was fixed with Flem- 
ming’s fluid and stained with safranin. This combination cannot be 
expected to stain bacteria. Failure to stain with Giemsa and methylene 


154 THE JOURNAL OF PARASITOLOGY 


blue does occur in certain bacilli which are common in the necrotic 
region of these sores. The stain is therefore not so reliable a cri- 
terion as location, uniformity of size, association with other free indi- 
viduals, etc. 

The time required for one complete multiplicative cycle is approxi- 
mated in the discussion of inoculation experiments. 

Schizogony in M. musculi would be expected if the schizonts con- 
taining spores had not been seen, since the peculiar distribution of 
the trophoplasts cannot be readily explained by any other kind of mul- 
tiplication. The smallest individuals are usually very numerous in 
localized regions and differ but little in size. Older stages occur in 
fewer and fewer numbers unaccompanied by the small forms, proving 
that they have migrated from the focus of the multiplicative process. 

Multiplicative reproduction in Myxosporidia was demonstrated by 
Cohn (1896) in M. lieberkuhni and by Doflein (1898) in Glugea 
lophi. Minchin (1903) describes fission and budding and refers to 
the multiplicative schizogony of Glugea lophii as a kind of schizogony, 
adding that “this kind of reproduction is probably very common, if not 
universal, in the tissue and cell-infecting Myxobolidae and Glugeidae.” 
Doflein (1911) makes provision in an outline of the life-cycle of a 
typical myxosporidian for schizogony and suggests it is typical as a 
preliminary to sporogenesis, but gives no specific illustrations and does 
not elaborate this as a process of multiplicative reproduction ee 
dent of the formation of sporoblasts. 

Laveran and Mesnil (1902) review the various methods of repro- 
duction in Myxosporidia, referring to budding as described by Cohn 
in Myxidium leberkuhni, also to binary fission as described by Doflein 
in Chloromyxum leydigi, and to the simultaneous division of the 
nuclei in the process of plasmotomie, but cite no typical cases of 
schizogony. 

The occurrence of multiplicative schizogony in a species of Myxo- 
bolus in the bile of the flounder has been observed by the writer. Plehn 
(1905) figures and describes a schizont with a large number of multi- 
plicative spores in Lentospora cerebralis from the salmon. It causes 
the so-called “twist disease” (drehkrankheit). He supposes that the 
spores develop into a cell which has a conspicuous nucleus that is lack- 
ing in the spores. In view of what has been learned about M. musculi, 
it seems more probable that Plehn’s nucleated cells are in the line of 
the propagative cycle. They are probably sporoblasts or gametoblasts. 
The non-nucleated spores are perhaps multiplicative spores. 

A schizont with ten multiplicative spores has been described by 
Nemeczek (1911) in Henneguya gigantea. 

It is now certain that the propagative cycle starts with a spore 
which is unlike the meront of the multiplicative stage. Beginning with 


HAHN—SPOROZOON PARASITES 155 


the spore, the staining properties of the propagativé trophoplast are 
distinctly different. In the paper already cited, Figure 14, Plate XX, 
represents a schizont with differentiated spores. They are probably 
not multiplicative spores as there stated. The latter are smaller and 
their nuclei do not stain. The propagative spores occur free in the 
myoplasm in fewer numbers, but with about the same pathological 
effects and habits as the multiplicative spores.. Because of the inti- 
mate and constant association of the small propagative spores having 
nuclei with large ameboid trophoplasts (Fig. 14), I have concluded 
that the former develop into the latter. This view might be less tenable 
if there was not a sharp limitation to the range of development which 
the parasites have attained in any given tissue. 

The fate of large propagative trophoplasts such as are shown in 
Figure 14 is probably some form of multiplication which results in 
moderate-sized sporoblasts. It is possible that they develop directly 
into primary sporoblasts, such as are undoubtedly represented in a 
trophic condition in Figure 5, and in a quiescent condition in Figures 
6 and 7. Such an interpretation conforms to the accepted life-his- 
tory for other species of Myxobolus. But if one is right in supposing 
that certain elongated spores of moderate size which have been occa- 
sionally. encountered isolated (Hahn, 1913: 113, Figs. 17 and 19, Plate 
XXI), and in small sporogenic cells (Ibid.: Fig. 16, Plate XX), are to 
be included in the life cycle of M. musculi, then it is difficult to recon- 
cile the stages represented with the sporogenesis as hitherto described 
by Mercier (1908), Keysselitz (1908), Schroder (1907, 1910), and 
others. There are apparently three different kinds of spores in M. 
musculi, One belongs to the multiplicative cycle and may without 
doubt be called an asexual type. The other two are very probably to’ 
be associated with the propagative cycle, and one may expect that they 
have some sexual significance. Of these, one is a spherical spore 2.5 
to 3u in diameter, which has a small well-defined nucleus and faintly 
staining protoplasm. They occur in large schizont cysts (Hahn, 1913, 
Fig. 14, Plate XX), and are produced in rather large numbers. They 
no doubt become the sporoblasts that are so numerous in tissue adja- 
cent to them in the one tissue where they have been encountered. The 
latter are identical to sporoblasts such as are figured in this paper 
(Figs. 5,6, and 7). The other type of propagative spore was encoun- 
tered in the same slide as the above and in the immediate vicinity of 
them. They are contained in cells having a diameter of about 12p. 
These sporocyte cells appear to be of independent origin. They occupy 
the space left by an atrophied muscle fiber. The contained spores are 
2.5 by 4p in size and have rather large nuclei. Each sporocyte contains 
. from four to twelve spores. 


156 THE JOURNAL. OF PARASITOLOGY 


The elongated type of spore not yet has been satisfactorily 
explained. If the spherical spores which contain a stainable nucleus 
are identical with what was assumed to be multiplicative spores, the 
elongated spores may prove to be sporocytes. I am not altogether cer- 
tain that the one tissue represented was not harboring a double infec- 
tion. A third hypothesis is that the small spherical spore is a micro- 
gamete and the larger elongated spore is a macrogamete. In this con- 
nection it is interesting to note that in the muscle fibers where typical 
medium-sized sporoblasts are abundant, occur also several small elon- 
gated cells with pointed, densely staining nuclei, having a terminal 
position (Hahn, 1913, Fig. 18, Plate XXI). One may suppose that 
these are motile microgametes, but at present no evidence is available 
to substantiate the hypothesis. 

One may conclude with reasonable assurance that the sporoblasts 
do arise from a very common type of spore which arises by a process 
of schizogony, and that the propagative sporoblasts are sufficiently 
differentiated from the multiplicative spores to be easily distinguished 
while yet in the schizont cyst. I believe that after a succession of mul- 
tiplicative cycles ending in multiplication by schizogony, there follows 
a schizogony which generates spores that become differentiated very 
early into either gametes or primary sporoblasts. (See also page 102 
in the first section of this paper for time relations.) 

The primary generative cells of M. musculi certainly do not arise 
by free cell formation in large myxoplasms, such as is the case in 
M. pfetffert of the barbel, and Sphaeromyxa labrazesi (Lav. and 
Mesnil), according to Schroder, 1907. The primary propagative cells 
of M. musculi, on the other hand, are set free simultaneously by one 
or the other of the schizonts described above. This conclusion is based 
not only upon the existence of two or more types of schizonts, but 
upon the fact that in four tissues where sporoblast stages occur, they 
are very numerous and at approximately the same state of development. 

The propagative stages have not been encountered so frequently 
as the multiplicative stages. This is probably due to the fact that they 
are not nearly so abundant. In some tissues one may find both kinds 
present, but according to my observations, one or the other is always 
greatly predominant. With the exception of the elongated spores 
which occur in certain small cysts that have been figured and described 
elsewhere (Hahn, 1913: 204, Fig. 16, Plate XX), there is no evidence 
that the multiplicative and propagative trophoplasts do not have prac- 
tically the same structure and appearance when small. 

There is absolutely no evidence that they are generated consecu- 
tively by budding or fission or plasmotomie, but. quite the contrary. The 
propagative stages gradually differentiate from the multiplicative type, 


and by the time one can positively identify them as such, they are dif-. 


HAHN—SPOROZOON PARASITES 157 


ferent both in appearance and staining reaction. When unmodified by 
the contraction of the muscle fibers, they are more or less spherical 
bodies with almost transparent glassy cytoplasm and a small vaguely 
staining nucleus (Hahn, 1913). Older conditions are shown in Fig- 
ure 18 of the paper just referred to. They have a large well-stained 
nucleus and fit loosely in the space which they have eaten in the myo- 
plasm. The shape varies from round to oval, and evidence of active 
mobility or of pseudopods is often lacking. Somewhat earlier stages, ° 
when compressed by the shortening of the muscle fiber, have long 
extensions of the cytoplasm (Fig. 5). The nucleus is also extended 
into a long slender mass and sometimes extends into fhe thicker por- 
tions of the protoplasmic branches. This condition does not seem to 
be quite normal. Many cases of less compressed myxoplasm occur as 
regularly distributed spindles. 

Besides very small sporoblasts, there are numerous good examples 
of larger sporoblasts and sporocysts in all stages of sporogenesis and 
sporocysts with immature and more or less mature myxospores. Stages 
not figured in the plate of this paper will be found in my paper of 1913. 

When unmodified by the contraction of the muscle fibers, the sporo- 
blasts are probably more or less spherical with a small nucleus (Fig. 7), 
or a large one (Fig. 6), and almost transparent vitreous cytoplasm. 
The nucleus does not stain intensely, but is more or less free from 
characteristic stainable bodies (Hahn, 1913, Figs. 18, 21 and 35, Plate 
XXI). Presumably these are the same stage of the organism as those 
which are encountered frequently in an ameboid condition fitting 
loosely into irregular transverse clefts of hypertrophied muscle fibers 
(Fig. 5). The conditions in some cases, such as Figures 6 and 7 here 
and Figure 18 (Hahn, 1913), suggest that there is an advanced condi- 
tion in which ameboid activity is lost. If so it is probably just pre- 
ceding the process of sporogenesis. There is a transition between the 
ameboid condition and the inactive condition wherein the myoplasm is 
divided into narrow transverse partitions by very numerous spindle- 
shaped cells which lie with their long axis at right angles to the length 
of the fiber. It is unsafe to say to just what extent the mechanical 
action of the muscle and the number of parasites are responsible for 
these alterations in shape. Thélohan (1891) figures and describes 
exactly the same condition in fish muscle fibers. He also interprets 
them as sporogenic cells. 

Sporoblasts are sometimes so closely packed in the space once occu- 
pied by a muscle fiber that, though the form of the fiber remains, the 
myoplasm can be seen only rarely (Fig. 18). When thus packed 
together, these cells form a pseudo-epithelium which can be distin- 
guished from a slightly degenerated epidermal or mucous epithelium 
with the greatest difficulty. Practically one must depend in many 


158 THE JOURNAL OF PARASITOLOGY 


cases upon a general resemblance to other epithelial masses in the 
same tissue, the cells of which have entered upon some easily recog- 
nizable stage of sporogenesis. Such pseudo-tissues are either more or 
less obscured by the hypertrophied myoplasm, muscle, and vascular 
nuclei, or are so closely packed that unless spread out mechanically in 
smear preparations, suitable specimens for drawings cannot be found. 
It is such a scattered group that was selected for the camera draw- 
ings represented in Figures 7 and 18. For purposes of reproduction it 
was necessary to exaggerate the detail of both nuclei ahd the cyto- 
plasm of the parasites. The disinclination to stain is still retained to 
a limited degree in the propagative stages. 

The epithelioid tissue just referred to must not be confused with 
another condition which has already been described (Hahn, 1913), in 
which the hypertrophied nuclei of vascular and connective tissue 
occupy the mold of a muscle fiber and, mingled with the remnants of 
the myoplasm, resemble a bit of degenerating epithelium. 

The identity of the cells of which these pseudo-tissues are com- 
posed rests upon very positive evidence. Not only can one easily find 
obvious differences between them and true epithelium, but"there are 
many such masses lying among the atrophied muscle fibers, many of 
which are in stages of sporogenesis like that represented here (Fig. 3). 
On a single slide one cannot fail to connect stages identical to those of 
Figures 3 and 6 (below) with the less obvious stages in Figures 7 and 
18. There are also interesting isolated groups of sporoblasts identical 
in appearance to those forming the epithelioid masses that occupy 
small spaces in the myoplasm (Fig. 6). Differences in the size of 
the nuclei are to be expected when it is recalled that we are comparing 
primary and secondary sporoblasts with pansporoblasts and possibly 
other stages of the propagative cycle. Figure 7 is magnified 560 diame- 
ters and Figure 6, 750 diameters. It is noteworthy that the group in 
Figure 6 is accompanied in the same fiber by a pansporoblast with ten 
or eleven nuclei. Between the former and the latter the hypertrophied 
myoplasm has lost the fibrillae. That the bodies represented in this 
fragment of muscle fiber were invading parasites is clearly obvious. 
The muscle hypertrophy alone is significant. Adjacent fibers have 
numerous isolated parasites, while the epithelioid masses and numerous 
stages of sporogenesis like Figure 3 are on the same slide from which 
the group in Figure 6 are taken. 

It is rather by analogy with other Myxobolus than by direct obser- 
vation that one must interpret the various propagative stages which 
have been encountered in the tissues of Fundulus. The majority of 
the older stages such as those in the pseudo-epithelium are probably 
sporoblasts. As already stated, those with large and small nuclei may 
prossibly be gametoblasts. There are some very large spherical stages 


HAHN—SPOROZOON PARASITES 159 


with two large and two Small nuclei from which the sporogenesis 
starts. With numerous succeeding stages leading up to Figure 3 one 
has, at least, ample proof that trophoblasts whose nuclei stain are 
destined to give rise to propagative spores, i. e., myxospores. 

It is of considerable interest that the early propagative stages like 
trophoplasts have a destructive career. Their scattered distribution 
in the younger stages is due to a rather extensive motility either upon 
the part of the parental schizont or upon their own activity. But when 
nearly mature they evidently become less active. The masses which 
occupy the mold of the muscle fibers suggest in a general way pseudo- 
cyst formation such as has been found in the gill (Textfigs. 1, 2, 
and 3), and is common in many of the other species (M. pfeifferi of 
the barbel disease). 

The pseudo-epithelium (Fig. 18) formed by the propagative stages 
of M. musculi is a most remarkable condition and deserving of more 
attention. The simulation of normal or slightly hypertrophied host 
tissues is a most deceiving circumstance. When a parasite having such 
qualities occurs in small numbers and more or less isolated, the most 
careful observer will fail to recognize it. Moreover, if a sufficient 
number of tissues is not available, suitable stages for a positive identi- 
fication will be wanting. The facts just noted are important because 
of their possible bearing upon the epithelioid tissues of mammalian 
cancer. Adami (1910) states that cancer tissue resembles nothing so 
much as a parasite upon the mammalian tissues. The propagative 
stages of M. musculi frequently give the appearance of a typical 
epithelioma. 

SUM MARY 


For the results of inoculation experiments bearing upon the life- 
history see the summary at the end of the first section of this paper. 
1. M. musculi has a series of multiplicative cycles starting with the 
myxospore, followed by a propagative cycle, ending in the myxospore. 

2. There are two or more types of schizonts and schizogony. 

3. Multiplicative reproduction is carried out by means of a large 
schizont which gives rise to a very numerous progeny of very minute 
spores. 

4. The multiplicative spores and trophic stages do not take up any 
stain thus far utilized, with one not very satisfactory exception. 

5. Multiplicative trophoplasts and schizonts migrate into unin- 
fected tissue, particularly just before the quiescent period preceding 
schizogony. 

6. All propagative stages possess a nucleus which reacts to basic 
stains. 


160 THE JOURNAL OF PARASITOLOGY 


7. The schizonts which give rise to primary propagative spores also 
migrate into new tissues before undergoing schizogony. 

8. Another process of schizogony exists in which the schizont is 
very large and the spores, though larger than multiplicative spores, are 
small and have a small nucleus which reacts to a basic stain. 

9. A third type of propagative schizogony may possibly exist in 
which the schizont is small and the spore very large, with a large 
nucleus which reacts to a basic stain. 

10. If the conditions in 8 and 9 are trustworthy, there is a differ- 
entiation of gametes into macro- and microspores. 

11. Sporoblasts, whether arising from conjugation or destined to 
conjugate, are ameboid, trophic, having the ability to migrate to a 
limited extent only when immature, and losing this property later. 

12. Multiplicative stages perforate muscle fibers extensively and 
bring about profound hypertrophy. Propagative stages while yet 
trophic are also predacious, but to a less degree. The latter give rise 
to characteristic irregular transverse clefts in the fibers. Such clefts 
vary in number, shape and size, and occur 11 more or less atrophied 
fibers only. 

13. The passive propagative sporocytes pass through all the char- 
acteristic stages of sporogenesis such as have been described for 
M. pfetfferi (Keysselitz, 1908). . 

14. Closely packed primary and secondary sporoblasts form an 
epithelioid tissue which at times has the appearance of integumentary 
epithelium and closely resembles mammalian epithelioma. 

15. Pseudocysts occur, having many myxospores in a common 
sporocyst plasm. They probably arise by the fusion of closely packed 
sporocysts. 


MYXOBOLUS PLEURONECTIDAE OF WINTER FLOUNDER 


A winter flounder (Pseudopleuronectes americanus) having open 
sores was collected by Dr. W. E. Sullivan in the vicinity of Woods 
Hole. When examined, the flesh proved to have undergone patholo- 
gical changes almost identical to what has already been described in 
Fundulus. The flounder was 8 inches long and had three lesions. 
One on the dorsal side was %4 inch wide and 1 inch long; the other 
two were smaller. The integument was either white and partially 
decomposed or completely gone. The underlying flesh was red and 
vascular at the surface and less transparent than normal. These 
external conditions resemble the appearance of the myxosporidian dis- 
ease of Fundulus as much as one could expect, considering the differ- 
ence in the integument, skin, and color of the flesh of these fish. 

Suitably stained smear preparations of the flesh present almost the 
same pathological conditions as are found in the fundulus disease. 


HAHN—SPOROZOON PARASITES 161 


There are present hypertrophied muscle fibers and epithelium cells, 
degenerated nuclei, mucus cells, and numerous bacteria limited to the 
most disintegrated parts. Numerous fibers contain considerable num- 
bers of unmistakable trophic stages of the multiplicative cycle of a 
Myxobolus. One could not distinguish these from the same stages of 
M. musculi of the Fundulus. Large multiplicative schizonts, almost 
mature sporoblasts, and myxospores are also to be found in the same 
tissues. With the exception of the myxospores, there is no noticeable 
difference in the propagative stages and those of M. musculi. 

The myxospores are not very abundant, but they are suitably 
stained for comparison with other species. One has no difficulty in 
distinguishing them from the myxospores of M. musculi by their shape 
(Fig. 2). The latter are tapered more at the polar end and the polar 
capsules are drawn out into a narrow apex. 

The flounder parasite has myxospores which are 14.84 long and 
11.9% wide. Those of M. musciuli are 14.3u long and 6.7 wide. 
Immature myxospores of M. musculi are 12 by 7.5y by actual measure- 
ment. In both cases the figures here given are the average of several 
different spores. The flounder myxospore has polar capsules which 
are 6u long by 3.7 thick, and the fundulus parasite has polar capsules 
6.5 by 2.0u. The flounder myxospore is therefore 24 shorter than 
M. pfeifferi and 1.9» narrower. In shape and appearance it resembles 
the latter closely. Allowing for slight variations in size and shape due 
to difference in maturity, the discrepancy between the myxospores of 
the two fish is too great to consider them as belonging to the same 
species. The inoculation of one host species by myxospores from the 
other will easily settle this question. In the meantime, the name 
M. pleuronectidae is proposed for the flounder parasite. It is probable 
that many species of the flatfish are subject to attacks by this parasite. 

It is interesting to note that one Chloromyxum myxcspore was 
encountered in the tissues of this flounder. 

The articles cited in this portion of the paper will be listed at the 
conclusion of the paper in the September number of the JouRNAL. 


162 THE JOURNAL OF PARASITOLOGY 


EXPLANATION OF PLATE 


Fig. 1—Five sporoblasts of C. clupeidae from the same slide as Figure 16. Note the 
unstained cytoplasm of the sporoblasts with thin filaments of host tissue residues separating 
one sporoblast from another. Compare this non-staining material with that in Figures 8, 11 
and 16. Note the square form of both myxospores and sporoblasts. The capsule nuclei are 
applied to the polar capsules in the right hand lower sporoblast. The sporoplasm is. unstained. 
The sporoblasts each contain a developing spore the nuclei of which are probably imperfectly 
stained. XX 1575 


Fig. 2—A myxospore of M. pleuronectidae from a lesion of the back of a winter 
flounder (Pseudopleuronectes americanus). 1575. 


Fig. 3—A sporoblast of M. musculi undergoing sporogenesis. The specimen here 
represented is one of many in a mass of cells similar to that in Figure 7, The dark border 
is stained serum. A space exists between the latter and the sporoblast, due to shrinkage. 
There are about 30 well defined nuclei. A few appear to be elongated as if about to 
divide. 560. 


Fig. 4.—Three trophoplasts of M. musculi from the eye muscles of an inoculated Fundulus 
that had died from a general infection of the head region. The parasite has not taken up 
any stain while the host tissue has. These three cases show as many stages in the hyper- 
trophy of muscle nuclei which the parasites have apparently attacked. Note the small 
trophoplast is associated with a nucleus showing normal alveoli while the larger trophoplasts 
are associated with nuclei from which alveoli have partially‘or completely disappeared. 1575. 


Fig. 5.—A fragment of an atrophied muscle fiber from a large open lesion of Fundulus 
containing propagative trophoplasts, possibly sporoblasts of M. musculi. I regard these as 
earlier than in Figure 6. The position of the long axis of the sporoblasts and their cavities, 
which is at right angles to the length of the muscle fiber, is due most likely to the contraction 
of the fiber. Compare the granular nuclei in this with Figures 6 and 7. 300. 


Fig. 6—A muscle fiber from Fundulus with several sporoblasts of M. musculi in the 
same cavity and one isolated. The many small nuclei of the latter indicate that it is 
in an advanced stage of sporogenesis. The large size of the nuclei in the others indicates 
that they are in a much later condition than those shown in Figures 5 and 7. Atrophy of the 
myoplasm is just beginning. 750. 


Fig. 7.—A group of sporoblasts of M. musculi at about the same stage of development 
as in Figure 5. The group lies adjacent to an epithelioid tissue which has replaced a com- 
pletely atrophied muscle fiber. These cells are drawn out of the mass sufficiently to permit 
drawing details which are not possible in the compact masses, one of which is shown in 
Figure 18. 560. 


Fig. 8—Portions of four muscle fibers from the dorso-branchial region (not body 
muscle) of the young of Clupea harengus which had numerous pseudocysts of myxospores of 
C. clupeidae. No myxospores occur in the head region. All the muscle is thus riddled with 
the trophoplasts of the Chloromyxum. They are both inter- and intra-fibrillar. When inter- 
cellular, note they have crowded the muscle fibers. Fibrillation and striation of the muscle 
fibers is entirely lacking. 300. 


. Fig. 9.—A portion of a muscle fiber from the body muscle of F. heteroclitus having a 
typical infection of very young multiplicative trophoplasts of M. musculi. 300. 


_ Fig. 10—Three mature myxospores of C. clupeidae showing the four polar capsules 
stained. 1650. 


Fig. 11.—Sporocyst of C. ieprie in an atrophied myoplasm from anterodorsal -body of 
muscle of young C. harengus. Compare with Figures 1 and 16. Note the increase in size. 
Sep tae plasm is unstained. Sporoplasm has assumed a more or less rectangular form. 


Fig. 12.—A multiplicative schizont of M. musculi in the myoplasm of an atrophied fiber 
from Fundulus. 750. 


Fig. 13.—A medium sized trophoplast of C. clupeidae migrating from an old tissue to 
new. X300. 


Fig. 14.—A large schizont of M. musculi from the same slide as Figure 12 which has 
not yet undergone schizogony. 560. 


; Fig. 15.—A large schizont of C. clupeidae from the body muscle of young Clupea harengus 
in which no pseudocysts are present and no myxospores were found. ‘These schizonts are 
abundant in comparatively normal muscle fibers. 300. 


_ Fig. 16.—Four sporoblasts of C. clupeidae from inflamed body muscle in the ventrolateral 
region. The two left-hand sporoblasts are enclosed in the sporocyst and the right-handed 


-sgaeerre are free. The latter are comparable to the shaded portions in Figures 1 and 


Fig. 17.—Photograph of a typical lesion in F, heteroclitus which afterwards proved to be 
caused by a typical infection of M. musculi. Note the swelling, the loosened and projecting 
scales, and the open central area from which the integument has disappeared. 


_ Fig. 18—A mass of sporoblasts of M. musculi giving the appearance of an epithelium. 
The truncated form of the mass is due to the fact that these sporoblasts have occupied 
the space left by the muscle fiber whose hypertrophy they have brought about. 300. 


PLATE 


CONTRIBUTIONS TO THE STUDY OF PARASITIC 
PROPOZOA,: | 11.* 


MYXOBOLUS TOYAMAI NOV. SPEC., A NEW MYXOSPORIDIAN PARASITE IN 
CYPRINUS CARPIO L. 


ROKUSABURO Kubo 


While studying Cnidosporidia in some fresh-water fishes during the 
last few months, my attention was attracted to a minute white spot on 
the branchial lamella of a Cyprinus carpio. Examination under the 
microscope showed that the white spot was no other than a round cyst 
of a myxosporidian containing numbers of ripe spores each having 
only one polar capsule. The fish that harbored the Myxosporidia wa’ 
a year old, having a length of about 6 cm. On searching carefully all 
the branchiae of the fish under the dissecting microscope, I found 
another round body situated near the free end of a branchial lamella, 
the diameter of which was about 200u. Since that time, many fishes 
of the same kind, and reared in the same pond where the above- 
mentioned infected fish had been found, have been examined for the 
same parasite, but it has not been found again. Consequently, the 
material is too scanty for detailed study. I will try, however, in the 
following pages, to give the results of observations on the one fish, 
which are probably of some interest, since the morphology, and espe- 
cially the life-history, of the unicapsulated Myxosporidia seem, so far 
as I am aware, to have been left in obscurity. 

The branchiae of the infected fish were cut into pieces, fixed with 
Schaudinn’s or Fleming’s fluid, imbedded in paraffin, cut in serial sec- 
tions of 2 to 4 thickness and stained with Giemsa’s solution or Heiden- 
hain’s iron hematoxylin, the latter being counterstained with eosin or 
orange G, 


MORPHOLOGY OF THE TROPHIC STAGE 


I expected that in sections would be found many young develop- 
mental phases of the organism which could not be observed externally 
in the fresh state, but in the study of the numerous sections, to my 
disappointment, only very few of the parasites were observed, showing 
that the infection in the present case was one of slight degree. The 
focus of infection was the connective tissue of the gill filament: The 
connective tissue became swollen by the infection of the parasite, and 
with its growth the tissue around it formed a thick layer, penetrated 
by numbers of capillaries (Figs. 1 to 3). A similar phenomenon has 


* From the Pathological Laboratory of the Imperial Sericultural Experiment 
Station, Nakano, Tokio, Japan. Director, Dr. T. Kagayama. 


164 THE JOURNAL OF PARASITOLOGY 


already been observed by Cohn in My.robolus minutus and by Schroder 
in Henneguya acerinae. The parasite in the branchiae is generally 
ovoidal in form (Fig. 1), but sometimes a calabash-shaped one is 
present (Fig. 2). This is probably caused by the union of two closely 
neighboring individuals. The gill-filament infected is not so greatly 
swollen as is the case with Henneguya acerinae, Myxosoma dujardini 
according to Thélohan (1895), and Henneguya gigantea according to 
Nemeczek. 

The youngest form found has an oval shape. The dimensions are 
about 67 by 50u, showing clearly the differentiation of the protoplasm 
into ectoplasm and endoplasm. The ectoplasm exhibits vertical stria- 
tions (Fig. 3), similar to those of Mystdiwm lieberkiihni, Myxobolus 
pfeifferi according to Thélohan (1895), of Henneguya acerinae and 
also of Sphaeromyxa sabrazesi according to Schroder (1907), and 
My-xobolus gigas according to Auerbach. LDesides this structure, in 
some specimens the ectoplasm differentiates very fine plasmic proc- 
esses, 2 to 3 long, from its surface (Fig. 3). Auerbach (1909) noted 
a structure analogous to the above-mentioned one in My-obolus fuhr- 
manni, but he could not determine whether it belonged to the parasite 
or to the tissue of the host. Schréder (1907) observed a similar differ- 
entiation of the ectoplasm in Sphaeromy.«a sabrazesi, stating that “an 
der Oberflache des Ektoplasms erkannte ich bei einigen Exemplaren 
einen zottenahnlichen, wenig ttber lu hohen Besatz.” 

The endoplasm has a coarsely granulated structure. The nuclei are 
round or oblong, in size varying from 1 to 4u. They are scattered in 
the endoplasm, unlike the nuclei observed previously by Thélohan, 
Schroder, etc., who found them situated rather in the middle portion 
of the endoplasm. 

In some young specimens, where the spore formation had begun 
to take place, I noticed that the nuclei and the pansporoblasts took a 
peripheral position, while in the middle portion a large round granu- 
lated body of distinct contour, but with no nucleus in it, was observed. 
I could not determine whether it is an accumulation of the endoplasm 
or an inclusion. 

In the older cyst, which is oval-shaped and of about 190p in maxi- 
mum diameter, the ectoplasm becomes thinner than in the younger 
form. In the periphery of the endoplasm, numbers of nuclei are to be 
found, and towards the middle portion of it matured spores and several 
developmental phases of pansporoblasts to spores. 


SPORE FORMATION 


The nuclei in the plasmodium may be distinguished as vegetative 
and generative. The latter are always found in a round cell which 
takes stains more deeply than the surrounding endoplasm. The uninu- 


KUDO—MYXOBOLUS TOYAMAI NOV. SPEC. 165 


cleate cells are the “sphéres primitives” of Thélohan (1895), “pan- 
sporoblasts” of Gurley, or “Propagationszellen” of Keysselitz (1908). 
The propagative cell is of oblong or spindle shape, though usually 
round in form, with dimensions of 4 to 8u. The nucleus is often situ- 
ated excentrically (Fig. 4). A caryosom, as Keysselitz mentioned, is 
always found in it. The propagative cell multiplies by division into 
two or three daughter cells (Figs. 5 to 16). These points correspond 
to some extent with those of My-.vobolus pfeiffert according to Keys- 
selitz (1908) and to Mercier, and Mywxidium bergense according to 
Auerbach (1912). The nuclear division in the propagative cell oi 
Sphaeromyxa sabrasesi according to Schroder (1907 and 1910), My.ro- 
bolus pfeifferi according to Keysselitz and to Mercier, and Henneguya 
psorospermica according to Auerbach, is reported to be mitotic. In 
the present form I also observed mitotic division. The chromatin, 
through the coil stage (Figs. 5 to 7), divides into two parts, exhibiting 
very often the central spindle (Figs. 8 to 10). In this respect it 
resembles that of Mysxidiwm bergense studied by Auerbach (1912). 

The propagative cells resulting from the multiplication go on to 
spore formation. The greater propagative cell (macrogamete) and the 
smaller one (microgamete) take an elongated form and associate with 
their lateral surfaces. At first a space is seen between them (Figs. 17 
and 18), and finally the cytoplasm of both cells fuses at the place of 
contact (Figs. 19 to 23). 

The association of two binucleate cells, observed by Schroder in 
Sphaeromyxa sabrazest and by Keysselitz (1908) in My.xobolus petf- 
fert, does not exist in the present parasite. ‘he association of the two 
uninculeate propagative cells in the present Myxosporidia strikingly 
resembles those observed by Mercier (1904) in Myxrobolus pfetfferi 
and by Auerbach (1912) in Myxidium bergense. But the nuclei of the 
associated form do not fuse into one, as Mercier thought happened in 
My.xobolus pfetfferi. 

The nuclear change in the pansporoblast coincides to some extent 
with that mentioned by Auerbach (1912) in Myxidimm bergense. 
Instead of uniting into one, the nuclei in the associated form undergo 
division. The smaller nucleus divides into two at the peripheral posi- 
tion of the pansporoblast, being destined for the nuclei of the pan- 
sporoblast (Figs. 23 to 26). The greater nucleus: repeatedly divides 
mitotically with the growth of the pansporoblast (Figs. 22, 25 to 31). 
In the fully developed pansporoblast, ten nuclei are observed, besides 
two nuclei of the pansporoblast and the reducing nuclei. At this stage, 
the contents of the pansporoblast separate into two sporoblasts, each 
of which contains five nuclei (Fig. 31). Of the five, two are found in 
a plasmic mass (sporoplasm in the later stage), one is in a cell which 
usually has a vacuole in it (nucleus for polar capsule and polar fila- 


166 THE JOURNAL OF PARASITOLOGY 


ment), and the remaining two are for the spore membrane. They are 
clearly recognizable in young spores, as is shown in Figures 32 to 37. 
When the spore is fully developed, the membrane of the pansporoblast 
is broken up, and the spores consequently become free in the endoplasm 
as in bicapsulated Myxobolus according to Keysselitz. As I mentioned 
above, we always recognize several developmental stages of the spore 
in the older cyst. 


MORPHOLOGY OF THE SPORE 


The spore has a pyriform shape, with a peculiar attenuated anterior . 
and broadly rounded posterior extremity (Figs. 38 to 45). It has no 
bilateral symmetry. The spore-membranes of lateral surfaces are 
usually curved in opposite directions (Figs. 39 and 40). The form 
agrees well with that of Myxobolus piriformis described and illustrated 
by Balbiani and by Thélohan (1895). Spores of the calabash shape, 
however, occur not infrequently in.the present case (Fig. 38). The 
spore-wall is comparatively thin and composed of two valves, superior 
and inferior. At the plane of junction the shell is somewhat thickened 
(Figs. 41 and 44). The surface of the spore usually represents no 
special structure. Very rarely a single, short, tail-like process about 
1.5y in length is seen at the middle part of the posterior end (Fig. 41). 
Thélohan (1895) observed a similar abnormality of the spore in 
Myxosoma dujardini and described that “quelques spores anormales 
ont un prolongement caudal.” I also regard the above-mentioned 
process in certain spores as an abnormality. The length of the spore 
is about 15y, the breadth 7 to 8 and the thickness 5 to 64. Thélohan 
gives the size of the spore of Myxobolus piriformis to be 16 to 18 by 
7 to 8u. In the fresh preparations one pyriform polar capsule is 
observed at the anterior half portion of the spore (Figs. 42 to 44), its 
dimensions beings 7 to 8 by 3 to 44. The wall is drawn out anteriorly 
into a minute duct which pierces the shell near its anterior extremity, 
affording exit for the polar filament. Thélohan did not measure the 
size of the polar capsule of Myxobolus piriformis. But it seems to be 
much smaller than the present form (compare his Figures 116 and 117, 
Plate IX, 1895, with my Figures 38 to 45). Auerbach (1909) observes 
spores with two polar capsules among unicapsulated spores of My-xo- 
bolus fuhrmanni. ‘In the present case, all spores have only one large 
polar capsule each, of which I will speak again when I come to the 
permanent preparations. Moreover, in some spores, the nucleus of the 
polar capsule is seen to be attached to it (Figs 42 and 43). The polar 
filament is easily extruded from the anterior end of the polar capsule 
when the spore is treated (Fig. 45) with a reagent like caustic potash, 
or hydroxyl, or pressed mechanically between the cover and slide 
glasses. The length of the filament measured after the spore has been 


KUDO—MYXOBOLUS TOYAMAI NOV. SPEC. 167 


freshly prepared and pressed agrees usually with the measurements of 
stained ones prepared according to my method (1913). The length 
of the polar filament of the parasite is 40 to 45, so it is 10 to 15 longer 
than that of Myxobolus piriformis measured by Thélohan. The pos- 
terior half portion of the spore is filled with sporoplasm. In fresh 
preparations, it is of a transparent, somewhat granular structure. 
Treated with iodin-aleohol, there appears a large vacuole stained 
brownish yellow in the sporoplasm. 

In fixed preparations, the anterior end of the spore-membrane is 
stained very faintly (Figs. 38 to 41). The duct of the polar capsule 
becomes easily visible. In some spores, close to the anterior end of the 
polar capsule, there is an oblong mass of protoplasm (Figs. 38 to 40). 
I took this structure at first to be a polar capsule and compared it with 
the “Korperchen” of Pimelodus blochi of Muller, 1. e., Myxobolus 
inequalis described by Gurley. But no such structure is observed in 
my present preparations, so that I cannot determine whether it is a 
degenerating polar capsule or some other structure. The nucleus of 
the polar capsule is always observed in young spores, well stained at 
the peripheral part of the capsulogenous cell (Figs. 33 to 37). In the 
sporoplasm, a large iodophile vacuole remains unstained, its diameter 
being about 3u. The iodophile vacuole of My-xobolus pirtformus 
observed by Thélohan is smaller than the present one. Two nuclei are 
always found in the sporoplasm situated closely to each other. They 
are usually of equal size (Figs. 37 to 41), but sometimes of different 
dimensions (Fig. 37). The position of the nuclei in the sporoplasm is 
not always the same. They are seen between the polar capsule and 
the vacuole (Fig. 39), on a lateral aspect of the vacuole (Fig. 38), or 
near to the posterior end of the spore (Figs. 40 and 41). 

To what genus and what species does the present parasite belong? 
Because of the presence of an iodophile vacuole, it is clear that it 
belongs to the genus Myxobolus. So far as | am aware, the unicapsu- 
lated Myxosporidia known up to the present time are four in number, 
all belonging to the genus Myxobolus: 


My-xobolus piriformis Thélohan 
Myxobolus unicapsulatus Gurley 
Myxobolus fuhrmannt Auerbach 
My-xobolus oculi-leucisci Trojan 


Of these, Myxobolus unicapsulatus is quite different from the present 
form. If one compares Figures 5, Plate 16 of Miiller (1841) with my 
Figures 38 to 41, one sees great differences between the spores. More- 
over, the habitat is quite different. Myxobolus fuhrmanni as stated by 
Auerbach (1909) was found in the connective tissue of the mouth of 
Leuciscus rutilus L. The spore is much larger than the present one 


168 THE JOURNAL OF PARASITOLOGY 


. EXPLANATION OF PLATES 


All figures except Nos. 1 and 2 are drawn with Abbe’s drawing camera. 
Figs. 1 to 41 from sections. 

Figs. 42 to 45 from fresh preparations. 

Staining: Figs. 7, 9, 14, 17, 18, 21, 22 and 35: Giemsa’s solution. 


All the others: Heidenhain’s iron hematoxylin and eosin. 


PLATES, I AND Tit 


Figs. 1 and 2.—Parts of longitudinal sections of infected branchial lamellae, 
showing the seat of the parasite. 1, * 160; 2, x 320. 


Fig. 3.—A peripheral portion of the parasite, showing the differentiation of 
the protoplasm. > 1000. 


Fig. 4—A propagative cell from the plasmodium. > 2250. 
Figs. 5 to 16.—Division of the propagative cell. >< 2250. 


Figs. 17 to 21—Association of the macro- and microgametes. > 2250. 
Fig. 22—Nuclear division of a macrogamete. 2250. 


Figs. 23 and 24.—The same of the microgamete. > 2250. 
Figs. 25 to 30.—Several developmental stages of the pansporoblast. > 2250. 


Figs. 31 and 32.—Segmentation of the pansporoblast into two sporoblasts. 
x 2250. 


Figs. 33 to 37—Young spores in development. 2250. 

Figs. 38 to 41. Matured spores. > 2250. 

Figs. 42 to 45—Spores from fresh preparations. 1000. 
Fig. 45—A spore with polar filament extruded. > 1000. 


PLATE I 


Si 


ie 


io 


\ 
See GR ce ee 


ete 


NM 


BO 5:5 Ore. 


PLATE II 


KUDO—MYXOBOLUS TOYAMAI NOV. SPEC. 169 
(length 18 to 204; breadth, about 8; thickness, 6u, and the length of 
the polar capsule, 9 to 102). The spore membrane is thickened at the 
posterior end and has 4 to 6 notches. None of these points agree with 
the observations mentioned. above onthe present Myxobolus. The 
same is true of My.robolus oculi-leucisci, which was found according 
to Trojan (1909) in the vitreous humor of the eye of Leuciscus rutilus 
L. Though the size of the cyst is almost equal to my parasite, the 
spore is smaller and different in structure. 

I have spoken only partially of the comparison between the present 
Myxobolus and My.xobolus piriformis, and will compare them here 
again in the following synopsis: 


My-xobolus piriformis 
EDaitateee =a - Branchiae and spleen of Tinca 


The present Myxobolus 
Branchiae of Cyprinus 


tinca L.; kidney of Misgurnus carpio L, 
fossilis 
2S i as “Les kystes branchiaux de cette Small round cyst in the 


connective tissue of the 
gill-filament 


espéce se reconnaissent a leur 
minceur: il forment de petites 
stries filiformes et non des 
tumeurs sphériques comme le 
M. ellipsoides” (Thélohan, 1895: 


348) 
Spore 

OGM: «el. 2 Pyriform, with attenuated ante- Pyriform, with attenuated 

rior extremity anterior end; often cala- 
bash form 
SIZE athaareatsi Length Breadth (Max.) Length Breadth Thickness 
16 to 184 7 to 8u 15u 7 to 8% 5 to 6h 

OMAR cos « Undescribed, figured only, capsule 7 to 8 by 3 to 4h 
seems smaller 

Todine 

vacuole....Smaller About 3 to 4% in diameter 


As will be seen from the above comparison, there are great differ- 
ences in the form of the cyst, the host, the size of the polar capsule, 
and the length of the polar filament, though the form and dimensions 
of the spore resemble each other. 

Hence I think the Myxobolus found by me is a new species, and 
propose to call it Myxobolus toyamai nov. spec. in honor of Prof. 
Dr. K. Toyama, who kindly introduced me to this branch of protozo- 
ology in the year 1909. 


REFERENCES CITED 


Auerbach, M. 1909. Bemerkungen iiber Myxosporidien. Zool. Anz., 34: 
456-65. - 

1912. Studien ttber die Myxosporidien der norwegischen Seefische und ihre 
Verbreitung. Zool. Jahrb., System., 24: 1-50. 

Keysselitz, G. 1908. Die Entwicklung von My-obolus pfeiffert Thél. 
f. Protistenk., 11 : 252-308. 

Kudo, R. 1913. Eine neue Methode die Sporen von Nosema bombycis 
Nageli mit ihren ausgeschnellten Polfaden dauerhaft zu praparieren und deren 
Lange genauer zu bestimmen. Zool. Anz., 41: 368-71. 


Arch. 


170 = THE .JOURNAL OF PARASITOLOGY 


1916. Contributions to the Study of Parasitic Protozoa. I. On the Structure 
and Life-History of Nosema bombycis Nageli. Bull. Imp. Sericult. Exp. Sta., 
Tokyo, 1: 31-51. 

Mercier, L. 1904. Contribution a l’études de la sexualité chez les Myxo- 
sporidies et chez les Microsporidies. Mém. acad. roy. Belg., 11: 1-52. 

Schréder, O. 1907. Beitrage zur Entwicklungsgeschichte der Myxosporidien 
Sphaeromyxa sabrazesi (Laveran et Mesnil). Arch. f. Protistenk., 9: 359-381. 

1910. Ueber die Anlage der Sporocyste (pansporoblast) bei Sphaeromyxa 
sabrazesi Laveran et Mesnil. Arch. f. Protistenk., 19: 1-5. 

Thélohan, P. 1895. Recherches sur les Myxosporidies. Bull sci. franc. 
Belg., 26: 100-365. 

Trojan, E. 1909. Ein Myxobolus im Auge von Leuciscus rutilus. Zool. 
Anz., 34: 679-82. 


Note. This paper was printed in Japanese in 1915 and is reprinted here 
at the request of the author. 


THE OCCURRENCE OF BOTHRIOCEPHALUS LIGULOIDES 
LEUCKART, WITH ESPECIAL REFERENCE 
TO ITS DEVELOPMENT 


SapAao YOSHIDA 
Pathological Department, Osaka Medical Academy, Osaka, Japan 


This larval cestode was first discovered by P. Manson at the post- 
mortem examination of a Chinaman at Amoy in 1882, and was 
described as Ligula mansoni by Cobbold in the following year. Since 
then about fifty-five cases have been reported, mostly from Japan with 
the exception of a few cases from Africa and the Malay Archipelago. 
The cases were all reported from the human kost and it has been ques- 
tioned for a long time whether this cestode larva was not confined to 
the human host. Some authors have suspected the existence of this 
worm in other animals without having actually proved its occurrence 
outside of man. A very few writers have described unsatisfactorily 
and uncertainly the occurrence of the cestode larva in question in 
animals. For instance, Dr. H. Miyake found twelve specimens of a 
cestode larva in the muscles of a monkey which had recently died, and 
he reported his belief that they were the same species as the liguloid 
larva from the human host, comparing them with the specimens and 
descriptions of previous authors. But the lack of a precise description 
for his own specimens prevented their positive identification. Other 
authors also, viz., A. Hirohata and J. Maejima, have proved experi- 
mentally that the larva is able to live and grow in the body of the rabbit 
by transplanting small pieces of the worm with a scolex into the body 
cavity of that host. 

In 1915, during my animal experiments with the encysted larvae of 
the lung distome, I accidentally came on August 11 across thirty-six 
specimens of this larval cestode in the body cavity and body wall of 
the cat employed in my experiment, which died in an extremely anemic 
and undernourished condition. Some worms were enclosed by a thin 
fibrous membrane, while others were lying free in the body cavity or 
in various tissues of the host. One, two, or even three worms were 
found in one capsule, and the latter were generally smaller in size than 
the former. The capsules lay in the muscular or subcutaneous tissues, 
varying in size and shape. A particularly large number of worms were 
found in the abdominal and pleural muscular wall, where they were 
tangled together into a ball or were creeping about here and there. 

Some portions of the body wall occupied by the worms had sup- 
purated. This agrees with the suppurating condition which is often 


172 THE JOURNAL OF PARASITOLOGY 


reported by various authors for human patients suffering from this 
cestode larva. Some worms were wound and twisted through various 
parts of body in such a manner that one end lay in the abdominal cavity 
and the other end in the abdominal wall, whereas the median portion 
of the worm lay irregularly in the body cavity and body wall. This 
state in the body of the host obviously proves the migratory tendency 
already frequently observed in the human host. 

' Fresh specimens were extremely mobile, especially in warm physi- 
ological salt solution, varying actively the shape and size of body. 
Large specimens measured 40 to 75 cm. in length and 17 to 20 mm. in 
breadth. There were also many other specimens in various stages of 
development or growth. Even in the same individual the length and 
breadth varies considerably according to the state of contraction. Gen- 
erally, when it was killed by a fixing agent such as a hot saturate solu- 
tion of corrosive sublimate, the worm contracted to two-thirds the 
length of living specimens. 

From my own observation of the morphological character and- 
anatomical structure of this worm, as well as its identification by Prof. 
Dr. Ijima, who was the first writer to describe this cestode larva in 
Japan, it is evident that the worm in question is the same as Manson’s 
larval cestode of man. Thus I have proved the actual occurrence of 
this cestode larva in an animal. Furthermore, I am inclined to believe 
that the normal intermediate host of this tapeworm should not be 
sought in a human being, but in another animal, though the parasite 
has not been found previously in the latter host while it has been found 
so often in the former. Why it seems to occur so often in the human 
body and so seldom in other animals doubtless depends upon the fact 
that human parasites are sought more carefully and are hence more 
frequently found than those of animals. It is obvious that the further 
development of this cestode larva would be impossible, or less likely at 
least, if the larvae were normally confined to the human body as a 
natural intermediate host. I am of the opinion that many animals, 
domesticated and wild, will be discovered to act as the intermediate 
host of this larval cestode. 

About six months after my discovery, M. Sugimoto in Formosa 
reported cestode larvae from the pig as Both. liguloides, and added that 
his specimens were quite similar to and probably identical with the 
specimens of Sparganum raillieti Ratz 1913 from the pig. The descrip- 
tions of Ratz’s and Sugimoto’s specimens indicate their likeness. But 
it is doubtful whether they are identical with Manson’s larval tapeworm 
from man or my specimen from the cat. 

As stated above, the great majority of the cases infected with this 
cestode larva have been reported from Japan. They were found in 
various districts throughout the country, but especially often near 


YOSHIDA—BOTHRIOCEPHALUS LIGULOIDES 173 


Osaka, the section where thirty-three out of fifty-five cases (60 per 
cent) are recorded. Thus I am now in a favorable locality to study 
the worm in question. Many difficulties, however, are encountered in 
studying the worm, because it occurs very rarely and consequently is 
found only accidentally in the human body or in other animals. In 
spite of efforts by various authors at various times, nothing is know1 
of the life-history of the parasite. 

I tried twice animal experiments to determine the final host of this 
larval cestode. In the first case I used two young cats as hosts and 
the cestode larva from the cat mentioned above. On August 11, 1915, 
two larvae were fed to one young cat and one larva to the other. The 
cats unfortunately died on the 17th of the same month from some 
unknown cause, and on dissection, no parasites of any kind were found 
in the alimentary tract. 

In the second case I made the experiment with the cooperation of 
my colleague S. Yamada. On June 26, 1916, a specimen of this cestode 
larva was obtained from a patient who suffered from the worm in the 
left side of her abdominal wall. The specimen measured 15 cm. in 
length and 5 mm. in breadth, and was enclosed in a capsule. This larva 
was given through a catheter to a young dog which was 27 days old, 
and had been reared in our anatomical departnient. Before the feeding, 
the feces of the dog were repeatedly examined for parasite eggs, and 
we found the eggs of Dipylidium caninum only. Afterwards we exam- 
ined daily for parasite eggs; and on the thirteenth day after experi- 
mental feeding we first found a new kind of parasite egg which 
increased in numbers day after day, and ultimately attained a maximum 
condition that continued until the animal was killed. 

The eggs are elongated oval in shape, tapering toward both poles, 
markedly sharper at the anterior than at the posterior end. On the 
anterior pole they are provided with a small operculum. They are 
mostly symmetrical, the curvature on the two sides of the long axis 
being unequal. One may find a minute globular thickening of the egg- 
shell at the posterior pole in some eggs, such as is observed in the eggs 
of certain distomes. The eggs closely resemble those of Dibothrio- 
cephalus, but are darker brown in color and different in shape. Some 
measurements in microns are as follows: 


1 2 3 4 5 6 7) 8 9 
Peneth: oo 5... 40-9 75.8 74.5 69.3 68.7 68.5 64.5 64.5 62 
ipteadthi 2.) ©43:) 37 33 S71 38.7 36.3 41.9 37 37 
WRatTOD setae: eet 204-1 9225-1 1.8621  1.77:0. FOB 5a 174 171 


On August 26, over two months after the feeding, we killed the 
dog and examined it for the parasites. We found a few specimens of 
Ancylostomum, two of Dipylidiwm caninum, and one large tapeworm 


174 THE JOURNAL OF PARASITOLOGY 


belonging to the genus Dibothriocephalus. This measured 2.5 m. in 
length and 12 mm. in maximum breadth, the maximum length of pro- 
glottis being 2mm. When alive it was very active and its length and 
breadth varied considerably, as is usual among cestodes. The posterior 
extremity showed a bifurcated anomaly, the body being divided into 
two halves near the median line, one half being 80 mm. long by 5 mm. 
broad, and the other half 50 mm. long by 3 mm. broad. The length of 
a proglottis in the bifurcated portion was constant (2 mm.). 

From observations on the external features and the internal struc- 
ture we easily identified this specimen as belonging to the genus Diboth- 
riocephalus, and bearing a close resemblance to Dib. latus. But I 
doubt whether the worm is identical with Dib. latus of the dog previ- 
ously reported. The known species of the genus Dibothriocephalus 
from the dog are Dib. fuscus Krabbe 1886, Dib. serratus (Diesing, 


Eggs of dibothriocephaloid cestodes from the various hosts. 620. 
A. From human host. B. From our dog used in experiment. C. From lion. 


1850), Dib. cordatus Leuck. 1863, and Dib. latus (L. 1748). The worm 
in question may easily be distinguished from any of the first three 
_ species mentioned above. 

L will add a few words on the comparison of this worm with Dib. 
latus which it resembles in some characteristics and not in others. 
Resemblance exists in respect to scolex form (though not accurately 
observed on account of irregular distortion by contraction), general 
form of the strobila, proportion of length and breadth of the proglottis 
in every part of the strobila, and general structure of internal organs 
and tissues. A remarkable point of difference is in the shape of the 
eggs. The eggs of this worm are easily distinguished from those of 
Dib. latus by their shape and the ratio of length to breadth. 

The eggs of the new worm are elongated oval in shape and mostly 
asymmetrical, the curvature on both sides of long axis being unequal, 
and they taper toward both poles, ending slightly pointed. The anterior 


YOSHIDA—BOTHRIOCEPHALUS LIGULOIDES 175 


pole is more pointed than the posterior, as stated above. The propor- 
tion of length to breadth, varying from 1.53:1 to 2.25:1, is greater than 
that (1.16: 1 to 1.48:1) of Dib. latus. The eggs of Dib. latus are oval, 
both poles ending equally rounded and relatively broader than those of 
the new worm. 

Measurements show that there is a great variation of egg size in 
Dib. latus according to the species of the host, whether human or other 
animal. The eggs of Dib. latus from other animals are the same in 
shape but much smaller than those from the human host. The next 
table serves to show the variation of egg: size. 


Human Human Human Lion Lion Lion 

1 2 3 1 2 3 
WMCTIE CHS sree oe a a arenieea 74.2 69.3 66.1 58.1 54.8 54.8 
Breaden 4 vsons sete | 51.6 46.7 50 35.4 35.4 33.8 
[iio] eae ee Ree tee tears 1.48 :1 S221 1.62 :1 1.54 :1 1.62 :1 


From the above tables it is evident that the eggs of the new worm 
are midway in size between those of the tapeworms from the human 
host and from the lion. 

In spite of such a great difference in the size of eggs, dibothrio- 
cephaloid cestodes from human host and other animals have been con- 
sidered to be the same species as Dib. latus by all previous authors. 
If this identification by the previous authors is unquestionable, the new 
- worm might be identified as Dib. Jatus and the remarkable difference in 
size and shape of eggs be considered a mere variation among the same 
species. If this supposition is right, one must reconsider the animal 
experiment. If the worm obtained from the dog under experimenta- 
tion is supposed to be Dib. latus, the dog must have swallowed a larva 
of this cestode species ; that is, have eaten the raw meat of salmon trout, 
which is considered in Japan to be the only species of fish harboring the 
larval form of this cestode. During our experiment the dog was always 
kept in a cage and fed regularly, so that he never obtained fish as food 
or accidentally. Before the experiment the dog was nursed by the 
mother or fed upon remnants of food which were generally boiled or 
roasted and could not be expected to contain a living larval cestode. 
Such a careful feeding experiment makes it impossible to think of an 
accidental infection with Dib. latus by food containing the larva. 
Therefore I am doubtful that the worm is surely identical with Dib. 
latus, and consequently it is a question whether the dibothriocephaloid 
cestodes from the human host and from other animals have been cor- 
rectly identified by previous authors. 

There is only one remarkable character, the egg size and shape, use- 
ful to distinguish positively the worm from the known species, Dib. 
latus. Egg size and shape of parasites, however, is generally assumed 


176 THE JOURNAL OF PARASITOLOGY 


to play an important role in determining species. Agreeing with this 
point of view supported by H. B. Ward, A. Looss, and other helmin- 
thologists, I have the following opinion in respect to the eggs: I am 
inclined to believe the worm in question will be experimentally deter- 
mined hereafter to be the matured form of Manson’s larval cestode 
and quite different from Dib. latus. Consequently, some cases — espe- 
cially from natural infection— of supposed Dib. latus from other 
animals such as the lion, dog, cat, etc., previously reported might have 
been mistakenly identified and really might have been the mature form 
of Manson’s larval cestode, or indeed of still another species. 

The natural mode of infection by Dib. latus also seems to support 
my supposition. To become infected with Dib. latus, it is necessary to 
eat raw fresh meat of fish harboring a larval form of this tapeworm; 
such fish are the salmon trout in Japan, or pike, salmon, perch, etc., in 
Europe. Generally the dog is fond of uncooked meat, but not of fish, 
in Japan at least; so it is unnatural and very rare for dogs to get fresh 
fish as food. Therefore it is puzzling to me why dogs are infected so 
often with this tapeworm in Japan, especially in the districts where the 
fish intermediate host is not found. The same difficulty holds good for 
the case of the lion, tiger, and other animals which are frequently 
infected with Dib. latus, although these wild beasts are accustomed to 
eat other weaker beasts and birds, but not fish so far as we know. 

It has already been proved, however, by previous authors that the _ 
plerocercoid larva of Dib. latus from fish can develop to the adult form 
in the alimentary tract of dog and cat. So it is probable, I think, that 
the dog and perhaps other beasts like the lion, tiger, etc., can become 
infected with two kinds of dibothriocephaloid tapeworms, viz., the well- 
known species Dib. latus, and a new species, the adult form of Manson’s 
liguloid larva. 

At any rate to have found the adult form of Manson’s larval tape- 
worm is both important and interesting, not only as a contribution to 
the knowledge of the development of the worm itself, but for the deter- 
mination of species of dibothriocephaloid cestodes from the human host 
and from animals. 

In closing, I wish to express my appreciation to Prof. Dr. Ijima, 
Chief of the Zoological Institute, Tokyo Imperial University, for his 
kind identification of the cestode larvae, and to Prof. Dr. Sakurane, 
Chief of the Dermatological Department of our hospital, for his cour- 
tesy in placing the material at my disposal. 


A FURTHER NOTE ON THE LIFE-HISTORY OF 
GONGYLONEMA SCUTATUM * 


Brayton H. Ransom AND Maurice C. HALL 


In two recent papers, one of which is an admirable monograph of 
the larval forms of the heteroxenous parasitic nematodes and the other 
a comprehensive study of the Gongyloneminae of North Africa, Seurat 
(1916: 739; 1916a: 358) has expressed the opinion that certain larval 
nematodes found in various species of coprophagous beetles (Apho- 
dius, Onthophagus) which we identified (Ransom and Hall, 1915: 154; 
1916: 80-86) as the larvae of Gongylonema scutatum probably belong 
to another species, G. mucronatum Seurat 1916. The adults of the 
latter species have been found by Seurat in the mucosa of the esoph- 
agus and base of the tongue of the Algerian hedgehog (Erinaceus 
algirus Duv.). It has not yet been recorded from sheep or cattle. 
Easily recognizable differences between G. scutatwm and G. mucro- 
natum are as follows, the statements relative to structural characters 
of the latter being taken from Seurat’s description: 

In G. scutatum the cervical papillae are situated about midway 
between the anterior border of the nerve ring and the anterior end of 
the body, each in the center of a rounded cuticular shield; in G. mucro- 
natum they are situated at the anterior third of the distance between 
the anterior border of the nerve ring and the anterior end of the body 
and are not inserted in the center of a cuticular shield. In G. scuta- 
tum the caudal pores are subterminal, in G. mucronatum situated at 
about two-thirds of the distance between the anus and the tip of the 
tail. In G. scutatum there is no papilla in the neighborhood of the 
vulva; in G. mucronatum an unpaired papilla is situated on the ventral 
surface of the body about 0.1 mm. behind the vulva. 

In view of Seurat’s opinion we have examined in detail numerous 
specimens of Gongylonema collected from the esophagus of sheep and 
cattle in various parts of the United States and have failed to find 
among them any corresponding to G. mucronatum, or to any species 
other than G. scutatum. If G. mucronatum is present in the United 
States, it is not likely that it occurs in sheep or cattle; at least, it must 
be rare in these hosts. Consequently, even considering the fact that 
we did not make a detailed microscopic examination of every worm 
from which eggs were obtained for feeding to insects in the experi- 
ments recorded in our former paper, but depended in many instances 


* From the Laboratory of the Zoological Division, Bureau of Animal Industry, 
U. S. Department of Agriculture. 


178 THE JOURNAL OF PARASITOLOGY 


upon the gross appearance of the parasites as sufficient for their iden- 
tification, it seems scarcely possible that there should have been 
invariably present in the material fed to the insects not only the eggs 
of G. scutatum, but also those of another species, whose presence in 
the sheep or cattle from which our material was obtained we con- 
stantly overlooked. The only apparent possibilities of error affecting 
our interpretations of the results of our experiments in feeding croton 
bugs and beetles in addition to the one just mentioned are (1) that the 
insects were already infested, and (2) that during the progress of the 
experiments they acquired the parasites from some other source than 
the material originally fed to them. In the case of the beetles we 
realized that some of them probably already harbored the parasites at 
the beginning of the experiments and gave due consideration to this 
probability in interpreting our observations ; but the possibility of such 
a circumstance in the case of the croton bugs is very slight in view of 
the fact that we have frequently examined croton bugs caught from 
the same places as those used in the experiments without finding 
Gongylonema larvae. The second possibility is also very slight in the 
case of the croton bugs, as they were kept during the experiments 
either without access to food or fed only on bread crumbs or similar 
food unlikely to contain nematode eggs. Furthermore, croton bugs 
kept in a similar manner and used in other experiments have never 
shown Gongylonema larvae. The beetles used in our experiments in 
some instances were kept in containers with unsterilized feces from 
sheep and consequently might have acquired their parasites from this 
source, but in certain instances the feces in which the beetles were kept 
and upon which they fed were sterilized. In the latter case, even with 
beetles already infested, one would be justified in considering as we 
did that the newly hatched Gongylonema embryos observed in large 
numbers a day or two after feeding, and the developing larvae in 
progressive stages observed later in due course of time, came from 
the eggs contained in the material fed to the beetles. So far as we 
are able to perceive after reviewing our records and recollections, our 
experiments in the feeding of Gongylonema eggs to insects were ade- 
quately safeguarded and controlled in all essential respects, with the 
exception that possibly sufficient care was not taken to exclude the 
eggs of species other than G. scutatum. With this possibility of error 
in view, slight though it is, the senior author has carried out a new 
series of experiments in feeding croton bugs. 

The insects used in the recent experiments were kept in flasks 
closed by cotton plugs. In addition to being supplied with material 
containing the Gongylonema eggs they were occasionally given fresh 
bread crumbs and a few drops of water. Numerous individuals caught 
from time to time in the same place as those used in the experiments 


RANSOM AND HALL—GONGYLONEMA 179 


were examined and found to be free from infestation. Some were 
also kept in flasks and fed bread and water, but no Gongylonema eggs, 
as controls against those fed Gongylonema eggs. The controls remained 
free from infestation. The Gongylonema material for feeding was 
obtained from a few infested gullets of sheep and cattle procured at 
an abattoir. All of the parasites were carefully removed from the 
gullets. By microscopic examination the fact was established that all 
of the worms present in each of the gullets corresponded to G. scu- 
tatum, special attention being given to the cervical papillae, absence of 
a postvulvar papilla, and subterminal location of the caudal pores. 
Female worms thus obtained and identified were washed in several 
changes of physiological salt solution to reduce the chances of foreign 
eggs adhering to their bodies, cut into small pieces, mixed with bread 
crumbs, and placed in the flasks containing the captive croton bugs. 
As in our former experiments, these croton bugs became infested with 
the larvae of Gongylonema. Individuals were examined at intervals, 
and various stages ranging from the newly hatched larvae up to the 
encysted forms, and exhibiting the same characteristics of structure 
as those described in our former paper, were recovered. An encysted 
larva taken from a croton bug seven weeks after eggs of Gongylonema 
scutatum were placed in the flask in which it was kept, measured 
1.9 mm. in length by 0.06 mm. in diameter. The pharynx was 0.035 
mm. in length, the esophagus 1.2 mm., its muscular portion 0.23 mm. 
The cervical papillae were 0.07 mm. from the anterior end of the body, 
the nerve ring 0.125 mm., excretory pore 0.21 mm. The anus was 
0.09 mm. from the tip of the tail, the caudal pores 0.025 mm. 

With reference to our experiments in feeding sheep with infested 
beetles and croton bugs (Ransom and Hall, 1916) it may be noted that 
their results if considered by themselves were less conclusive than those 
of the experiments in feeding Gongylonema eggs to insects, because 
of the small number of animals used and the lack of complete control 
of all the conditions which might have affected the experiments. 
Furthermore, no attempt was made to obtain a series of steps in the 
development in sheep between the larva and the adult. As a matter 
of fact in our former paper we did not insist upon the conclusiveness 
of the sheep-feeding experiments and considered them of importance 
only when viewed in the light of other evidence without which they 
would have been much less significant. Even though the experimental 
evidence that the larvae of Gongylonema scutatum in dung. beetles 
develop to maturity in sheep and other suitable mammalian hosts when 
the insects are ingested by these animals is less complete than that as 
to the development of the larval stage in the insects, such evidence as 
we have is in exact accord with that hypothesis, which moreover by 
analogy is strongly stipported by the known facts in the life histories 


i180 THE JOURNAL OF PARASITOLOGY 


of other parasites, and we are justified in assuming until very definite 
evidence to the contrary is brought forward, that sheep, cattle, and 
other suitable host animals become infested with Gongylonema scu- 
tatum as a result of swallowing infested insects, under natural con- 
ditions probably various species of dung beetles. 

From the foregoing it is evident that the validity of the results of 
our work on the life history of Gongylonema scutatum has not been 
affected by the question raised by Seurat regarding the correctness of 
our identification of the larval nematodes which we found in copro- 
phagous beetles and under experimental conditions in croton bugs. It 
is also evident that the nematodes found by Seurat (1916: 739, Fig. 5; 
1916a: 315, 346) in several species of Blaps, and because of the sub- 
terminal position of the caudal pores considered by him to be the larvae 
of G. scutatum, cannot belong to this species, unless it is a species 
whose larvae are characterized by an unusual degree of polymorphism. 
Whether the nematodes occurring in various species of coprophagous 
beetles in Algeria, which are strikingly similar to those which we have 
shown to be the larvae of G. scutatum, belong to G. mucronatum as 
Seurat (1916a: 317, 346, Fig. 11) supposes, remains to be determined. 
The basis upon which Seurat identified them as G. mucronatum is the 
location of the caudal pores at a considerable distance from the tip 
of the tail, a character in which they agree with the adults of this 
species. Clearly, however, apparent similarities in details of structure 
are not sufficient in the absence of other evidence to justify definite 
conclusions as to the specific identity of larval and adult nematodes, 
and further investigations will be necessary before the larvae described 
by Seurat as such can be accepted as the larvae of G. mucronatum. 
Because of their close agreement in structure with the larvae of 
G. scutatum, it is quite probable that they actually belong to this 
species, the adult stage of which Seurat has found to be common in 
Algeria. 

SUMMARY 

Despite the doubts raised by Seurat in recent publications, the con- 
clusions expressed in our former papers on the life history of Gongy- 
lonema scutatum are still valid. It has been definitely proved that 
dung beetles and croton bugs fed upon the eggs of G. scutatum become 
infested with an encysted larval stage of the parasite, and the evidence 
is very strong, if not quite conclusive, that sheep, cattle, and other 
suitable mammalian hosts become infested as a result of swallowing 
infested insects (usually under natural conditions, various species of 
dung beetles). 

The nematodes found in several species of Blaps in Algeria and 
identified by Seurat as the larvae of G. scutatum belong to some other 
species. 


RANSOM AND HALL—GONGYLONEMA 181 


It is not improbable that’the nematodes found in Algerian beetles 
which Seurat has considered to be the larvae of G. mucronatum in 
reality belong to G. scutatum. 


REFERENCES CITED 

Ransom, B. H., and Hall, M. C. 1915. The Life History of Gongylonema 
scutatum. Jour. Parasit., 1: 154. 

1916. The Life History of Gongylonema scutatum. Jour. Parasit., 2: 80-86. 
[Issued Jan. 29.] 

Seurat, L..G. 1916. Sur les gongylonémes du Nord-Africain. Compt. rend. 
soc. biol., 79: 717-742; figs. 1-5. 

1916a. Contribution a l’étude des formes larvaires des nématodes parasites 
hetéroxénes. Bull. scient. de la France et de la Belg., (7) 49: 297-377; figs. 1-14. 


NOTES 


The intermediate host of Schistosoma mansoni in Venezuela is discussed 
in a recent important paper by Drs. Juan Iturbe and Eudoro Gonzalez. By 
infection experiments with the mollusks of the valley around Caracas, the true 
intermediate host was found to be Planorbis guadelupensis. The miracidia of 
S. mansoni developed into sporocysts within this host, and ‘subsequently 
furcocercous cercariae were obtained from it. By immersion in water infected 
with these cercariae and by feeding experiments white mice were brought to 
develop adult S. mansoni. The paper is illustrated with two microphoto- 
graphic plates. 

A recent number of Japanese Medical Literature states that Schistosoma 
japonicum is reported by Narabayashi to depend on a small snail in the rice 
fields as its intermediate host. According to Pilsbury this snail should properly 
be called Blanfordia nosophora Robson. The cercariae invade the skin even if 
the latter is only damp. A definite relation exists between schistosomiasis and 
a skin disease called “kabure.” 

The same journal reviews a paper on Paragonimus westermanu in the 
Korean Medical Society Journal in which Muneta records from an autopsy 
the abundant occurrence of the fluke cysts in the abdomen under the peri- 
toneum; liver, spleen, heart, sternum and the cheek were also invaded. Some 
nodules contained adult worms; others did not. 


Davainea formosana, a new human tapeworm from Formosa and Tokyo, 
is described by Akashi in the Journal of the Formosa Medical Society and 
abstracted in Japanese Medical Literature. The specimens came from chil- 
dren. The species may be distinguished from the other member of the same 
genus long known as a human parasite by the following characters: 


Davainea formosana D. madagascarensis 
Length of bedy...2 «2.0: .7.20640.em. Z5 tO OD Gm. 
INUIMber On JOIntses- aces eee Over 700 500 to 700 
Elooks on suckers. -- anes elec None Armed : 
Adultisesments a... ecrise ree 2.0 to 2.5 by 10 mm. 2.0 by 1.4 mm. 
Boommasseseneee cee oeieerece 300-400 120-150 
Size of egg masses........... 0.26 by 0.13 mm. 0.3. mm. 
S1Z€ (OfNeES OS HNs. wate ie oe ee 99 by 46u 40h 
Size of onchosphere.......... 12 to 14% 15H 


“ESCHINORHYNCHUS MONILIFORMIS” IN NORTH AMERICA 


My attention has been called to the fact that in the Proceedings of the 
Philadelphia Academy (1874:76) is recorded with brief comments an exhibit 
by H. C. Chapman of specimens of Echinorhynchus moniliformis from the 
alimentary canal of the fox squirrel (Sciurus vulpinus). Stiles and Hassall 
also in their Preliminary Catalog of the Parasites, etc., (1894: 352) list the 
species from Sciurus niger as found in the Leidy Collection. The statements 
in my note must be corrected in accordance with these facts. No data are 
given in these brief records to determine whether the authors mentioned above 
had before them the true European species or the North American form which 
I have studied. Ee eBS Ve 


INDEX TO VOLUME Ill 


PAGE 
Animal Parasites of Man, by Fantham, Stevens, and Theobald (review).. 139 
Arthropoda, Observations on Polycystid Gregarines from................. 65 
Ascaris triquetra Schrank in Dogs, A Case of the Occurrence of......... 39 


Attacus cynthia Drury, Note on a Species of Nosema Infecting........... 136 


Book Reviews, see Reviews. 
Bothriocephalus liguloides Leuckart, The Occurrence of, with Especial 


Rererencer torts a evelopment ss emi as. sis cic os: devs c cicte sre dcayaie Sd ve bane 171 
Brookover, Charles: Diptera in the Human Intestine (note).............. 141 
Case of the Occurrence of Ascaris triquetra Schrank in Dogs............ 39 
masta Ges bhenGercariae Of INatal see fc acted ccc ccihs csetie + owe selene s 131 
eee ame eA AN ea eee tea a 5 via cuales vided ote docs vevesdecees 131 

of the Bitter Root Valley, Montana, Notes on the......:.............. 105 

Notes on Two Free-Living Trematodes from North America......... 10 
Cestodes from the Spotted Sting-Ray, Notes on Two.................... 34 
Contributions to the Study of Parasitic Protozoa. II. Myxobolus toyamai 

nov. spec., a New Myxosporidian Parasite in Cyprinus carpio L...... 163 

III. Notes on Myxosporidia Found in Some Fresh-Water Fishes of 

Japan, with the Description of Three New Species............... 3 
Cooper, A. R., see Job, Thesle T. 
Crab, Helice tridens (de Haan), On a Trematode Larva Encysted in a.... 76 
(Te, Gose (wae) anes Bae iodine. 6 doe CO RBC ae ee net rere 142 
Cynomys ludovicianus, Cytoleichus penrosei, a New Arachnoid Parasite 

Found in the Diseased Lungs of a Prairie Dog....... Bree pocere at iare 82 
Cyprinus carpio L., Myxobolus toyamai nov spec., a New Myxosporidian 

EPS (UGaetiemepe Srey cee eee e rem cies cities © os inte sale sYo 3% s' Giasiss'cltde eeisesisiels 163 
Cytoleichus penrosei, a New Arachnoid Parasite Found in the Diseased 

Eungs of a Prairie Doe? Cynomys. ludovicianus.......2.66 cc ceccee t's 82 
Dauercystformation of Trichomonas intestinalis..........000e cece eee e cece 28 
ERTUILE a OME SLM (Dad TLOEC)) TAM RAR Tete ofeloue ie = Sie ie i le's.o as 0,0 [sls/e eel diel sieveinsmyerave pacts 182 
Development of Gregarines and Their Relation to the Host Tissues: (1) In 

SEGROMOOCE: URGENT \NIETEO Te ok 58 GS ROE EEL BORA ECE enor iar: 124 
Mipterawiii tue Peli ae ENtESEINE (NOL) ecto soc svt os ws es tases oclsielee elles 141 
“Echinorhynchus moniliformis” in North America (note)............. 141, 182 
Effects of Radiation on the Development of Trichinella spiralis, with Respect 

to Its Application to the Treatment of Other Parasitic Diseases...... 43 
Endamoeba buccalis. I. Its Multplication and Periodicity................. 143 
Faust, Ernest Carroll: Notes on the Cercariae of the Bitter Root Valley, 

WY IGTRIIB on oe oe oho Ao ld ABE BS BRIO eae ee Bee errr. to coors tirrer seri 105 
Fishes, Notes on Some Nematodes from Fresh-Water................ ee YA 
Fundulus, Further Observations on Myxobolus musculi from.......... 91, 150 
Further Note on the Life History of Gongylonema scutatwm............+. i Fats 
Gongylonema scutatum, A Further Note on the Life History of........... Wi, 
Gregarines: Development of, and Their Relation to the Host Tissues: 

(1) In Sitenophora lactaria Watson...........0e2eerccccecconccccncs 124 


irom Artarapoda, Observations. on Polycystid. 2... ccc cscs se cece « 65 


184 INDEX TO VOLUME VII 


Hahn, C. W.: On the Sporozoon Parasites of the Fishes of Woods Hole 
and Vicinity. JI. Further Observations on Mysxobolus musculi from 
UNS oe aia acre lee la sv nse ao os ss seis Siva one Cals aioe cise eee 
II. Additional Observations on Mysrobolus musculi of Fundulus and 

a Nearly Related Species, M. pleuronectidae of Pseudopleuronectes 
DUN CV NGOS trie oo cw ai chee Os ow ond ov cla Sse sence he eee 

Hall, Maurice C., see Ransom, Brayton, H. 

Harvard School sotairopical) Medicine’ (ote) aay.05. heen eee ECE 

Helice tridens (de Haan), On a Trematode Larva Encysted in a Crab.... 

Honeij, James A.: see Tyzzer, E. E. 


Bouse Bly; Dangerous, (note): <2 s.6c2 hewn ds we see eee ee oe eee 


Ishiwata, Shigetane: Note on a Species of Nosema Infecting Attacus 
ENINEIUE OUETIEY 5c 5 6: 5ic050>»/ CGS aoe Ba Scene tara gs fe oR 


Japan, Notes on Myxosporidia Found in Some Fresh-Water Fishes of, 

with the Description of Three New Species:nc0.s00cen- seek cee 
Japaneses Medical diterature (reviews)! -o- nee eee eee eee ee eee 42, 
Job, Thesle T., and A. R. Cooper: Notes on Porocephalus globicephalus 


Kamm, Minnie Watson: The Development of Gregarines and Their Rela- 
tion to the Host Tissues: (1) In Stenophora lactaria Watson........ 
See also Watson, Minnie E. 

Kudo, Rokusaburo: Contributions to the Study of Parasitic Protozoa. 
Il. Myxobolus toyamai nov. spec., a New Myxosporidian Parasite in 


CYPTIRUS SCOTPI0 Las... 6's i3.s dinar ieh One ERI ee Oa ee ae 
Ill. Notes on Myxosporidia Found in Some Fresh-Water Fishes of 
Japan, with the Description of Three New Species.............. 
Leishmania brasiliensis (note)... eee ees pee see eee 
Life History of Gongylonema scutatum, A Further Note on the............ 
Linton, Edwin: Notes on Two Cestodes from the Spotted Sting-Ray.... 
ihe, Max. (note) iis.cicre. ceive ae,tc te eee eee eee cee eee 


Lynch, Kenneth M.: Dauercystformation of Trichomonas intestinalis.... 


Magath, Thomas B., see Ward, Henry B. 
Man, Parasites of: 


Animal Parasites of Man, by Fantham, Stephens, and Theobald (review) 
Dauercystformation of Trichomonas intestinalis..............ceeeccees 
Davatnea formosana: (note). .v.cc.s ¢. 200s +. 0 os ee oe eee 
Diptera in the’ Human. Intestine <(note),....c0:02 4) nn eee eee 
Endamoeba buccalis. I. Its Multiplication and Periodicity............ 
Japanese. Medical Isiterature\(feview)= <..:...«-<.. tees eee 
Medical and Veterinary Entomology, by William B. Herms (review)... 
Occurrence of Bothriocephalus liguloides, with Especial Reference to 


Its: Developmient sans): »,ocide istgs otro ar-torhent «oy sie4's bon [eee 
Medical and Veterinary Entomology, by William B. Herms (review).... 
Michigan University Biological Station’ (note)............. .20se nee eee 
Myxobolus musculi from Fundulus, Further Observations on......... 91, 


toyamati nov spec., a New Myxosporidian Parasite in Cyprinus carpio L. 
Myxosporidia, Notes on, Found in Some Fresh-Water Fishes of Japan, 


with the Description of Three New Species.............0.esete semen 
Nematodes from Fresh-Water Fishes, Notes on Some..............ee-- 
Nosema Infecting Attacus cynthia Drury, Note on a Species of........... 


Note on a Species of Nosema Infecting Attacus cynthia Drury.......... 
INGE oie sis wae w nce 21s toate Me ate LT 42, 90, 141, 


PAGE 


3 
140 
138 


124 


136 
182 


INDEX TO. VOLUME: IH 185 


PAGE 
PIGtGRNGIN LOLOL ED AGES ICUICC IES y 5.) Gon Fv cuchla alone = sane o 4 ble sides onele 138 
Some Nematodes from Fresh-Water Fishes................0ceeccceee 57 
the Cercariae of the Bitter Root Valley, Montana..................... 105 
Ewo Cestodes from the Spotted Sting-Ray.................0--<5--50% 34 
Two Free-Living Larval Trematodes from North America............ 10 


Nowlin, Nadine: Endamoeba buccalis. 1. Its Multiplication and Periodicity 143 


Observations on Polycystid Gregarines from Arthropoda.................. 65 

Occurrence of Bothriocephalus liguloides Leuckart, with Especial Refer- 
ACEC er SDC Vel OP men terncrset nit. 2-5 6 ms, vis ole tiene © Meee erin # via sine. s ee go where 171 

On a Trematode Larva Encysted in a Crab, Helice tridens (de Haan).... 76 


On the Anatomy and Relationships of Some North American Trematodes 21 
On the Sporozoon Parasites of the Fishes of Woods Hole and Vicinity. 
J. Further Observations on Myxobolus musculi from Fundulus....... 91 
II. Additional Observations on My-robolus musculi of Fundulus and 
a Nearly Related Species, M. pleuronectidae of Pseudopleuronectes 


Sea Ee aoa din jo aio ww ae a'e'w, «ss SeiN ed ors Siac ans ein eb eee 150 
Paragonimus westermani, New Human Tapeworm (note)............... 182 
RALLATE DUS EISE PEM SES 1 IOLE) ie sty s wlapn es «8 ab, ciaia's a isve's aye clbisials as ne 0 2 oie wale ee 182 
Rap AGalas GLODIEEPHAIUS NOLES OM, < 0.56 ~ 52s one < geccd esse wane evened eens 138 
Protozoa, Contributions to the Study of. II. Myxobolus toyamai nov spec., 

a New Myxosporidian Parasite in Cyprinus carpio L................. 163 
III. Notes on Myxosporidia Found in Some Fresh-Water Fishes of 
Japan, with the Description of Three New Species............... 3 


Radium Emanations: 
Effects of Radiation on the Development of Trichinella spiralis, with 
Respect to Its Application to the Treatment of Other Parasitic 


IDVSERSES conch adosgeGednaud 72O St Conn BSo er nee aeren ten an err 43 
Ransom, Brayton H., and Maurice C. Hall: A Further Note on the Life 
EIMSEOLVSO Le GONGUIONCING) SCULOIUIN oc. ceieies von cs vee saeco vede cen scees 177 
Reviews: 
Animal Parasites of Man, by Fantham, Stephens, and Theobald........ 139 
DS PELC SSE LOS eed Se ee 42, 140 
Medical and Veterinary Entomology, by William B. Herms........... 90 
eS AU AMICM IE CTIOLE) ON coe c's a © 3 ioc eid cnc cn cnce seuss scene wee aes om 182 
ELAS CLIVE (CTIENEE) ) PMY eee = cere) wc Soc ao ise vias force niale 2 sinisinw oldteyareiainrg 182 
Stenophora lactaria Watson, The Development of Gregarines and Their 
RetdtOnmtOmtaem El OStamiSSMGS 4 GL) Li... 5 - ncisis tu ners omles sewn anne 124 
Sting-Ray, Notes on Two Cestodes from the Spotted...................- 34 
Stunkard, Horace W.: On the Anatomy and Relationships of Some North 
AMIR@EIGtIN “LETT EIGUIES Joe Wien SES CORE EROS EEE Een Daas ee ac re Aes a 21 
Trematode Larva Encysted in a Crab, Helice tridens (de Haan), Ona.... 76 
Trematodes from North America, Notes on Two Free-Living Larval..... 10 
on the Anatomy and Relationships of Some North American........... 21 


Trichinella spiralis, The Effects of Radiation on the Development of, with 
Respect to Its Application to the Treatment of Other Parasitic Diseases 43 


Trichomonas intestinalis, Dauercystiformation Of..............00ee cece eee 28 
shropicamemMedicine ws hanvard School of (note)... ..¢. 0. cose. - weenie 42 


Tyzzer, E. E., and James A. Honeij: The Effects of Radiation on the 
Development of Trichinella spiralis, with Respect to Its Application 


fo.the Preamene of Other Parasitic. Diseases. «. 2.2m i.0. 5.62 ec seen 43 
Walton, A. C.: A Case of the Occurrence of Ascaris triquetra Schrank 

te ee oh PSS 6.0, oc a. Wo. + 5 > nein ol eacimin ioral aque ole bas oie ect i AOD. 
Ward, Henry B.: “Echinorhynchus moniliformts” in North America (note 

SRAM PRE RIRGNY ( oP aoc) tu)o ho ayein > 3.00,0 wn a eS es ee eee aatere ee 141, 182 


Notes on Two Free-Living Larval Trematodes from North America.. 10 


Sen. ee 


,, ‘ BN “ { his i * | i bt v" é : * . 
AY A pee i Ms : 
ee | a0 ys: are ny 
: - j i “Y iy A a) 
186 — INDEX TO VOLUME III ; 


i ae 

Ward, Henry B., and Thomas B. Magath: Notes on Some nena 
from Fresh-Water Pushes... 5:34 a aes eee mae 

Watson, Minnie E.: Observations on Polycystid Gregarines from Arthropoda 
See also Kamm, Minnie Watson 

Weidman, Fred D.: Cytoleichus penrosei, a New Arachnoid Paras Found 
in the Diseased Lungs of a Prairie Dog, Cynomys Iudovicianus...... 


Yoshida, Sadao: Occurrence of Bothriocephalus liguloides Leuckart, with — 
Especial Reference to Its Developments). 20s)... ) ee ee ae, 


On a Trematode Larva Encysted in a Crab, Helice tridens (de Haan).. 


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