16 DR. G. C. CHATTERJEE: THE CULTIVATION OF TRYPANOSOMA, ETC. lilled with bluish-stained granules. No other structure could THE CULTIVATION OF TRYPANOSOMA be made out. The posterior end (that opposite to the OUT OF THE LEISHMAN-DONOVAN aagellum end) was finely drawn out as it were into a small flagellum. BODY UPON THE METHOD OF In the moist hanging-drop preparation made on the third CAPTAIN L. ROGERS, I.M.S. lay the appearance of the fully developed parasite was most interesting. In examining the specimen under 1/16 th inch BY G. C. CHATTERJEE, M.B., oil immersion lens I found several elongated flagellate ASSISTANT BACTERIOLOGIST, MEDICAL COLLEGE, CALCUTTA. bodies moving slowly across the field by a lashing side-to- side movement of the this end the front (With Coloured Illustration.) flagellum, being of the moving parasite. The movement was distinctly slow, much slower than that of an FOLLOWING the method of L. ordinary’ trypanosoma my teacher, Captain Rogers, (trvpanosoma Brucei or Evansi). No wriggling movement I.M.S., for cultivating Leishman-Donovan bodies in citrate could be made out. The thick flagellum end was ’distinctly of sodium solution I have succeeded in developing trypano-. seen moving among the broken-down red corpuscles which soma from the Leishman-Donovan bodies. were pushed out by the jerking movement of the flagellum. In one field .I a a The patient from whom the blood for making the culture found group of these parasites lying in clump, their anterior ends being free, reminding one was taken by spleen puncture was an inhabitant of Sylhet. of the rosette formation of trypanosoma Lewisi by Laveran He gave a history that three members of his family had died, and Mesnil.2 The free ends showed the movement of the recently from fever with enlarged spleen and oedema of the flagella. In the free ones no movement in the posterior end legs. He had been suffering from the fever for six months of the parasite was seen. The are in the and was anæmic. The reached down to the following peculiarities noticeable parasites very spleen described above: the of the micronucleus at the was There was no (1) position umbilicus but the liver not enlarged. anterior end of the body of the parasites ; (2) the long, marked pigmentation. The temperature of the patient at thick anterior flagellum ; (3) the absence of (or, if present, the time of the puncture was 102° F. rudimentary) undulating membrane; and (4) the rudimentary On account of the of these The following procedure was adopted for taking the blood posterior flagellum. presence from the patient and for cultivating the parasites found in it. characters they would appear to be less closely allied to the class such as Brucei and The skin over the region of the spleen was cleaned with soap trypanosomidæ, trypanosoma and water and then washed with a 1 in 20 carbolic lotion. trypanosoma Evansi, than to the class of trypanopis is characterised A five cubic centimetre syringe, sterilised in a 1 in 20 carbolic described by KeysSelitZ,3 which by the of a micronucleus at the anterior a lotion and washed in boiling water, was used for puncturing possession end, long anterior its from the micro- the spleen. After drawing out the blood it was emptied thick flagellum taking origin a and a into a test-tube containing one cubic centimetre of 5 per nucleus, rudimentary, undulating membrane, hardly cent. citrate of sodium (the test-tube with the citrate solu- discernible posterior flagellum which takes no part in the tion being previously autoclaved at 115° C.). The test-tube movement of the parasite. In I to indebtedness to wsa then placed in a beaker which was put inside an ice-box. conclusion, beg express my Captain A stained slide preparation of the blood made on the first Rogers, officiating professor of pathology, Medical College, for me the and for day showed a good number of distinct Leishman-Donovan Calcutta, kindly allowing to publish paper bodies, from three to four being found in a single field. On revising it. DESCRIPTION OF THE PLATES. the next day a stained specimen made from the citrated Plate 1 is drawn from a crowded field blood showed increase of the in so comparatively showing great parasites number, numerous developed Leishman-Donovan bodies on the third day much so that from 200 to 500 bodies could be counted in a stained with Leishman’s stain. Drawn under a 1/12 th inch oil immersion single field instead of the three or four seen on the first day. lens. No fully developed body is seen here ; several dividing flagellated parasites as well as several elongated Leishman-Donovan bodies with- The showed increased they were dis- ’ parasites development; out any flagella are seen. in was tinctly larger size. The micronucleus distinctly Plate 2.-From the same specimen, selected from several fields. increased in size and was less circumscribed and was No. 3 is a fully developed parasite curled upon itself. elongated in shape. The micronucleus was situated at one Calcutta. end of the oval body instead of being in the middle. The contents of the body of the parasite were more granular; numerous dividing ones were seen. No flagellate forms were CHILD STUDY AND THE TREATMENT OF found on this day. PARALYSIS IN CHILDREN. On the third day the stained slide from the blood showed 1 a remarkable appearance. In many of the fields were seen BY ALFRED A. MUMFORD, M.D. LOND., M.R.C.S. ENG. elongated Leishman-Donovan bodies with a thick flagellum. HONORARY PHYSICIAN TO THE GREENGATE DISPENSARY, SALFORD. In some portions of the field they were seen in thick clumps, the clumps consisting of from 30 to 40 flagellate bodies. THE art of walking and of standing as carried on by a In these clumps were seen all the stages from a simple human seems at first to ba Leishman-Donovan body to a fully developed trypano- fully-developed being sight soma-like body. Several dividing ones were also seen hardly more difficult of acquisition than the swinging of a possessing two flagella, two macronuclei, and two elongated jointed pendulum. On more careful examination, however, bodies to each other but lying parallel possession ;;- each stage of movement and each position of rest are found a micronucleus situated transversely to the body. single to be gained only as the result of many trials and not a few Several bodies were found which were indistinguishable from fa which have a of an ordinary Leishman-Donovan body except by the possession ilures, placed complex arrangement many of a long flagellum. Besides these there were the fully de- muscles under the control of the will. It is difficult to over- veloped ones, in which the body was elongated (much longer estimate the fact that each additional accomplishment is than a Leishman-Donovan body) an n rrow and provided only acquired after much individual effort of the child with a long thick flagellum. The flagellum was longer than and is not the mere exercise of an inherited nervous the body of the parasite and was thicker than the flagellum or, inisation. The intricate nature of the muscular arrange- of an ordinary trypanosoma (trypanosoma Brucei or Evansi). ment is clearly brought out when by a partial paralysis of It took its origin from the micronucleus situated at one end the limb some constituent element is thrown out of action. of the parasite ; in many the micronucleus was connected to Defective or discordant movements then result. A similar the flagellum by a portion of darkly stained protoplasm. complexity is noticeable in the movements of the upper as of A vacuole-like organ was seen near the micronucleus ; in the lower limb. This is evidenced by the fact that in some it was found towards the middle of the body of the extension of the hand on the forearm we not only contract parasite. The macronucleus was found situated in the the extensors but almost, though not quite, simultaneously middle of the body and was elongated in shape and not so we also contract the flexors of the wrist with a little less defined as in the Leishman-Donovan body. No undulating I vigour. By this means we provide not only a break which membrane could be made out. The body of the parasite was 2 Doplein and Sv. Prowazek : Handbuch der Pathologischen Mikro- organismen von Wassermann und Kolle, S. 933, fig. 45. 1 The detailed description of the flagellate bodies given here is 3 C. Keysselitz : Uber Trypanopis Grobbenii, Archiv für Protiste confirmatory of that already noticed by Captain Rogers. kunde, S. 367.