ON A FIVE FLAGELLATE TRICHOMONAS

(N. Sp.) PARASITIC IN MAN.* ? By G. C. CHATTERJEE, m.b.,

Medical Collnje, Calcutta.

Flagellates inhabiting the intestinal tract

;u*e, as is well-known, very common in the

tropics. They have been made the subject of study in the Philippine Islands by Stilt and others and in Ceylon by Castellani and Chalmers. The Indian representatives have not, so far as

one could judge from published literature, re-

ceived much attention, except for the solitary instance by D. D. Cunningham who worked before the discovery of modern fixing and stain-

ing methods. This is probably due to the

supposition that the Indian intestinal flagellates are the same as those found elsewhere. But as

by studying by recent staining methods, I

found this supposition to be incorrect, more than one variety being found, altogether new to

science, I thought it advisable at present to

describe the one which I studied a little more in detail.

The flagellate in question has got five anterior

flagella, one posterior " rand" flagellum, one

undulating membrane?characters possessed by none of the intestinal flagellates hitherto des-

cribed. It was found by me in stools of six cases of chronic dysentery. The patients showed no

peculiarity, except in one marked anaemia of

secondary type was found. The stools were

^chylous, and frothy containing some mucus.

In three of them amoeba showing nucleus of the histolytica type were found. In two, besides

amoeba and the flagellates, numerous Lamblia

mtestinales were found.

Description of Living Specimens.

These are very actively motile, moving by Tuick jerky movement, through the fluid portions. ^ hen gliding through solid faecal matters, the

movement partakes of amoebic character. In

changing their direction while moving through semi-solid portions of the field they often double

themselves, the front end being parallel to

the posterior end. The Lamblia which were

found in the same fluid showed no such peculiarity. They are bigger in size and are much more active. The body of the former is pear shaped, one side being a little more convex than the other. In the convex side is seen, when the movement of the organism is slowed down a little, the wavy border of the undulating membrance, the undulations starting near the anterior end, going through a series of short curves and then ending in the tail end. In the anterior end, in slowly moving organisms, a slight depression can be made out from which a bunch of whiskers can be seen projecting out which move spasmodically forward and backwards like the oars of a boat. The posterior end terminates in a stumpy tail which shows only a side to side movement, when the organism is moving actively.

Description of Stained Specimens.

The specimens were fixed in Schaudin's fluid, then stained by Heidenham's Iron-ha3motoxylon method. A few specimens were made by adding a drop of 10 p.c. alcoholic solution of Iodine to a

drop of faeces placed on a slide and then quickly smearing it on the slide and then staining by Giernsa stain. This method I found gave very

good results, the nucleus, the basal granules, the

flagella, and the undulating membrane being clearly differentiated. The following structures were seen in stained

specimens?nucleus is very big, rounded in shape, a nuclear membrane is clearly seen. Karyosome is present in the form of small clumps distributed

irregularly through the substance of the nucleus. In many specimens nucleus is homogenous?no karyosome can be made out. The nucleus is

situated in the extreme anterior end just behind

the basal granules. The flagella are six in number, five being free

and directed forwards. The sixth one is a " rand"

flagellum forming the border of the undulating membrane, through the length of the body-?and then becomes free, projecting out near the tail.

Of the five free flagella, four are equal in length and are often found together, being directed forwards. The fifth one is slightly smaller in size and is

often separate, being directed to one side. In

many specimens, however, five are found to-

gether. The body of the parasite is pear shaped, in

most specimens one side is a little more convex

than the other. The anterior end is pointed, and just to one side of this pointed end is

a small depression around which are seen. origi- nating the anteriorly directed flagella (Fig. 1). this depression is a cystotome?110 regular cyto- Received for publication 29lh August, 1914?15d.

THE INDIAN MEDICAL GAZETTE. [Jan., 1915.

pharynx has been clearly made out. Just near

the depression are brightly stained granules (basal granules). From the basal granules are seen starting a fine chromatic line passing trans-

versely to the opposite edge surrounding the

cystotome. The " rand " flagellum is seen origi- nating from near the origin of the other flagella (the basal granule and the chromatic line), passes obliquely to the edge, and then along the

edge to a certain distance in a series of curves, and then becomes free near the tail end. In most

specimens the undulating membrane is clearly seen as a thinly stained wavy border, from near

the anterior end down to the tail. In Grieinsa

stained specimens the membrane is hidden by the

darkly stained body of the organism. In well

fixed specimens stained byGiemsa, a well marked

axostyle can be seen arising from the posterior part of the nucleus ending near the tail. In

several a full size red corpuscle was found in-

side the body; no vacuole is seen. In some por- tions of the field rounded forms were found.

These show a large oval nucleus at one side, basal granule, an indentation in the edge near the

basal granule (cystotome), and a bunch of flagella (often five in numbers) originating from the

basal granule. Undulating membrane and the

tail are not seen.

Besides, dividing forms were seen, but only rarely (Figs. 9 & 11) no encysted forms were seen.

Measurements of the Organism.

The body of the organism measures 8 to 10/m in length, 5 to 6^ in breadth, the 4th anterior

flagella 8 to 10a the short 5th anterior flagellum G to 8 fi. The projecting portion of the 6th " rand" flagellum is 3 to 4ju. The nucleus is

3fi in diameter. Systematic.

It evidently belongs to the family Tetramitidse. In habits and characters it resembles in every respect a Trichomonas. The possession of five free flagella is the only hindrance to its being included in the Genus Trichomonas?Trichomonas

having been defined by Doflien and Parisi and others as an organism possessing three to four

flagella. Recently finding of an organism by Alexeieff possessing four anterior flagella and one posterior

" rand " flagellum has caused Parisi, who found a similar or identical parasite, to sub-divide the Grenus Trichomonas into three sub-genera:?

(1) Trichomonas sensu stricta?with three anterior flagella and one

" rand " flagellum and an undulating membrane.

(2) Tricliomastix?with three anterior flagella and one free posterior flagellum which act as a

"

Schleppgeissel." (3) Tetra-trichomonas?with 4 anterior flagella,

one posterior " rand" flagellum bordering the

undulating membrane. Mackinon has added to these another sub-genus termed Tetra-tricho-

mastix* in order to include the organism found by her. The characters of this sub-genus are as

follows :?four anterior flagella, cne free posterior flagellum acting as a

"

Schleppgeissel." Following the precedence of Parisi and Mac-

kinon, the organism under consideration should be classed under a fifth sub-genus under genus Tri-

chomonas, which may be provisionally named Pentatrichomonas. For this purpose the defini-

tion of genus Trichomonas has to be changed to

one possessing three to five anterior flagella instead of one possessing three to four flagella. The

locality in which this parasite has been found will give its specific name, as Pentatrichomonas

Bengalensis. The classification will be as

follows :?

Class-Mastigophora-Diesing. Sub-ctass-Flagellata-Cohn and Biitsehli. Order-Polymastigina-Biitsclili. Family-Tetramitidse-Biitschli. Grenus-Trichomonas-Donne.

Sub-genus-Pentatrichomonas (N. Sp.). In conclusion I like to mention that flagellates

and ciliates are fairly commonly found in the intestinal contents in this country. True Tri- chonomas intestinalis (three anterior flagellata) have not as yet been met here. I have found

recently in three cases a Microstoma.f Whether

this pentatrichomonas represents Trichomonas

of other countries or only accidentally found in

particular type of cases remains to be seen. I

do not like to discuss the pathogenic property of

the new organisms?whether these are in common with ordinary Trichomonas simply Saprophytes or like Difremus?an intestinal flagellata found by Grabel?is pathogenic, remains to be seen by fur- ther observations. Prowazeck and Werner have

described a Lamblia and Chilomastix causing enteritis.

Descriptions of Plates.

The specimen was prepared by making a smear of stool, fixing in Schaudin?s fluid while wet and

tlien staining in Leisliman's stain and mounting in Hoyer's fluid. At no stage of the process the

preparation was allowed to dry. Drawn under Camera Lucida, 3 mm. apochroma-

tic lens and 18 eyepiece and tube drawn to 16 mm.

Fig. No. 6 shows a parasite in the process of

plasmolysis, Here the parts are very clearly seen. 1, 2, 3, 4, 5 are five free flagella, a?the flagellum bordering the undulating membrane ; b?bound- ary line of the body of the parasite; c?is basal granule; d?axostyles originating from nucleus.

* This distinction between Trichomonas and Trichomastix? the former having an attached posterior flagellum and the latter a free one?though accepted by most protozoologists has been reversed by Martin and Kobertson who figure their Trichomonas gallinarum as having four free flagella, whereas lYicliomastix gallinarum as having three anterior flagella and one posterior flagellum attached to undulating membrane. f This Macrostoma differs from the one described by Wen-

yon as Macrostoma Mesuili in many points. I intend to pub- lish notes on this later on.

ON A FIVE FLAGELLATE TRICHOMONAS (N. Sp.) PARASITIC IN MAN.

Hv G. C. CHATTERJEE, m.b.,

Medical College, Calcutta.

FIG 1 FIG 2

FIG 5

FIG 6 D

FIG 7

FIG 8

9j, S./..OAS,

FIG 1 FIG 2

F16 3 FIG 4

FIG 5

FIG 6

FIG 7

FJG 8

FIG 9

FIG 10

fig 11

Jan., 1915.] "QUININE PROPHYLAXIS.

Fig. 1. Shows an organism in which the

undulating membrane is clearly seen. There are

seen four out of five free flagella. Axostyle is

clearly seen. Fig. 2. Also shows four free flagella. No

axostyle is seen. Fig. 3. Shows five free flagella, and a

" rand "

flagellum. The undulating membrane is not

clearly seen. Two axostyles are seen. Fig. 4. Same as fig. 3. Fig. 5. Shows five free flagella, one

" rand "

flagellum, one axostyle. Fig. 7. Five free flagella. The flagellum border-

ing the undulating membrane seems to be free. Fig. 8. Five free flagella are seen. The undu-

lating membrane with its attached flagellum is seen at the lower part of the organism. The clear

portion bounded by a line at the upper portion seems to be due to plasmotysis of the organism.

Fig. 10. A dividing form?the two nuclei, two groups of basal granules and axostyles and flagella are seen.

Fig. 11. Seems to be a dividing form with two nuclei and six free flagella.

Postscriptum (Dec. 1914). At the time of correction of the proofs, I came

across a review in the October number of the

Tropical Diseases Bulletin of a paper by Derrien and Raynand under the title of Dysenteric chro- nique d Flarjelle Noveau in the July number of Bulletin Society Pathologye Inclica. In this it

appears the authors have found a new intestinal

flagellate which they descij bo under the name of Hexamastix Ardiu-Delteili. They found this parasite in a patient suffering from diarrhoea in the medical clinic of Mustafa in Algeria. The parasite is characterized by the following charac- ters measurement 10 to 15/7, long, 9 to 13/x broad. Nucleus and Blepharoplast are present. Close to the Blepharoplast arose an undulating membrane, terminating in a flagella, as well as five flagella varying in length 10 to 17/j. There was also ^n axostyle. This parasite is allied to Tricho- monas intestinals, but is distinguished by the presence of fine flagella. From the above description it seems that this

parasite resembles in many essential points the

?ne described by me in my paper, but the absence of the original paper which has not as yet arrived in Calcutta, it is very difficult for me to make out whether this parasite is the same as found by me or differs from it in minor points.

Inferences.

Stilt, E. R.?-A Study of the intestinal parasite found in Lavite provinces?Philippine Journal of Science, Vol. VI, P-210. .

?

Castellani and Chalmers?Note on an intestinal flagellate lr> man. Philippine Journal of Science, Vol. 5.

Castellani and Chalmers' Manual of Tropical Medicine, 1910. Cunningham?On the development of certain microscopic

organisms occurring: in the intestinal canal. Quarterly Journal of Science, Vol. XXI, 1881.

Doflein?Lehrbuch der Protozoenkunde, II. AufL 1909. Parisi, B.?Su alcuni flagellate Endoparasite, Archives do

Protistenkunde, Vol. XIX, p. 232. Alexeieff, A.?Trichomonas a Quatri Flagelles Antericurs.

Comptes des Seances et Memoires de la Societe de Biologie (Paris XXVII, p. 712). Maclcinon, D. L.?Studies on parasitic protozoa ; Quarterly

Journal of Microscopical Science, Vol. 59, p. 459. Martin and Robertson?Further observation on the ca;cal

parasites in fowl. Quarterly Journal of Microscopical Science, Vol. 57, p. 53. Gabel, M.- Zur Patliogenitat der Flegellaten-Ein fall

von Tetramitidendiarrhce. Archive de ProtistenUunde, Bd. XXXIV, S. Prowazek, S. V. and Werner, H.?Zur Kenntnis der sog.

Flagellaten Beihefte zur. Archive fiir Scliiffs und Tropen- hygene Beilieft, 5, p. 155.