The isolation of a Leishmania from 5.8% of 138 geckos (Hemidactylus turcicus) examined by culture of cardiac blood in two kala azar study areas in the southern Sudan is described. On morphological and antigenic grounds, this strain is designated as Leishmania hoogstraali sp. n. The work of Heisch (1958) and MansonBahr and Heisch (1961) in Kenya, as well as that of others, has suggested that reptilian Leishmania species should be considered in kala azar epidemiological studies. Leishmania adleri was isolated from the lizard Latastia longicaudata revoili during search for a vector of kala azar in Kenya. This species, which is antigenically more closely related to L. donovani than to any other mammalian species (Adler and Adler, 1955), produced local induration in man when inoculated intradermally or subcutaneously. The induration persisted for a month, and the parasite could be recovered up to a week after inoculation. The possibility of positive leishmanin skin reactions resulting from infection with such parasites under natural conditions was suggested. One facet of kala azar investigations recently carried out in southern Sudan by the United States Naval Medical Research Unit Number Three (NAMRU-3) was an appraisal of the possible significance of reptilian hemoflagellates in the epidemiology of the human disease. During these studies, in January and FebReceived for publication 22 September 1964. * From Research Project MR005.09-1603.9, Bureau of Medicine and Surgery, Navy Department, Washington, D. C. The opinions and assertions contained herein are the private ones of the author and are not to be construed as official or reflecting the views of the Navy Department or of the naval service at large. This study was supported in part by Research Grant E-4823 from NIAID, NIH, United States Public Health Service. A summary of this paper was presented at the First International Congress of Parasitology, Rome, in September, 1964. t School of Public Health and Tropical Medicine, University of Sydney, Australia, and Guest Investigator, Department of Medical Zoology, United States Naval Medical Research Unit Number Three, Cairo, Egypt, U.A.R. and Malakal, Upper Nile, Sudan. ruary 1963, the writer isolated from the gecko, Hemidactylus turcicus (Linnaeus), a Leishmania which is now described as a new species. The epidemiological significance of this and of another reptilian hemoflagellate found in the study area will be discussed in a future publication. MATERIALS AND METHODS Geckos (Hemidactylus spp.) were collected from the thatched roofs and walls of native tukls (huts) in two areas 100 miles apart, where kala azar has been intensively studied by NAMRU-3. These areas, in the vicinity of Paloich (32?32' E, 10?27' N) and Malakal (31039' E, 9031.5' N), Upper Nile Province, have been described by Hoog traal et al. (1962, 1963) and by Van Peenen and Reid (1963). Each gecko was prepared for cardiac puncture by lifting a small area of skin over the heart with fine forceps and removing the flap with heated scissors. After further cauterization of the exposed ar a, a sterile glass capillary, drawn to a point, was introduced into the heart for withdrawal of blood. Approximately 5 mm3 of blood was inoculated into individual tubes of Tobie's (1950) diphasic medium. The eventual mortality from a single cardiac puncture was about 20%. A thin blood film was prepared from each gecko and stained with Giemsa after fixation in methanol. Tissues were not examined for leishmanial forms. Slopes of the medium used were prepared in Cairo by adding 2.5 ml of base containing 10% of defibrinated rabbit blood together with 700 units crystalline penicillin and 700 ,ug streptomycin sulphate per ml into screw capped tubes. These were airfreighted to southern Sudan, the time in transit being about 2 weeks, and on arrival were stored in a refrigerator until used. An overlay of 1.5 ml of sterile Locke's solution containing similar concentrations of antibiotics as the base, was added to each tube before use. A low-temperature incubator was not available at the period of this investigation and cultures were held in an air-conditioned room in which temperatures ranged between 18.9 C to 27.7 C, with an average daily variation of 5.6 C. Cultures were examined for flagellate forms after about 7 days and at 21 days.
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