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The present study was conducted to Assess the efficacy of various diagnostic techniques for that all the suspected cases of buffaloes coming to TVCC, U.P. Pandit Deen Dayal Upadhyaya pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura were examined by parasitological and serological methods. (viz. giemsa stained thin blood smear, buffy coat method and TELAT) for the diagnosis of trypanosomiasis in naturally infected buffaloes. The order of decreasing diagnostic efficacy during present investigation was found as: TELAT (52.02%) > Buffy Coat method (45.62%) > Giemsa stained thin blood smear (38.20%).