Comparative efficacy of different in vitro cultivation media for Trypanosoma evansi isolated from different mammalian hosts inhabiting different geographical areas of India
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The present study was undertaken to establish an optimal medium for primary culture initiation and maintenance of T. evansi isolated from different mammalian hosts of diverse geographical regions of India viz. donkey/1 (Hardoi, Uttar Pradesh), donkey/2 (Junagarh, Gujarat), pony/1 (Hisar, Haryana), camel/1 (Bikaner, Rajasthan) which represented isolates 1, 2, 3 and 4, respectively. Primary cultures were initiated with all four isolates in five different in vitro cultivation media with seeding density of 1 × 106 trypanosomes/ml. The parasites of all four isolates could remain viable only for 48 h in medium E (Alsever’s solution) and for 72 h in medium A, C and D. Parasites reached to a maximum density (2.5–3.75 × 106/ml) within 24 h and thereafter, a sharp decline (0.5–0.75 × 106/ml) in the next 72 h was observed in 1, 2 and 3 isolates cultured in medium B. In isolate 4, parasite counts got more than doubled in 24 h and then decreased gradually up to sixth day post initiation of cultivation which thereafter increased gradually up to 34 days and a constant parasite number of 105/ml could be achieved for 90 days in medium B. During this prolonged culture the trypanosomes retained their long slender morphology and infectivity to mice.
KeywordsCulture media In vitro cultivation Livestock Trypanosoma evansi isolates
The authors wish to acknowledge their gratitude to the Director, National Research Centre on Equines, Hisar, India for providing research facilities to carry out the present study. The authors would also like to thank Mr. R. K. Dayal for technical assistance provided during the investigation. The financial support from Indian Council of Agricultural Research, New Delhi under National Fellow Scheme for the present study is duly acknowledged.
All authors certified that the animal experimentations were carried out in the present work after approval of Institute Animal Ethic Committee and as per guidelines set by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Animal Welfare Division, Government of India.
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