This study was undertaken to detect Trypanosoma evansi infection in clinically ill cattle, buffaloes and horses that were suspected of ‘surra’ infection. Various diagnostic tests were utilized in order to assess the prevalence of infection in the clinically ill and ‘suspected’ animals. Animals in the Eastern Region of India were systematically studied for T. evansi infection, which had not been tested for previously. Examination of Giemsa-stained blood smears detected 5·3%, 9·4% and 40·6% infections; the mouse inoculation test detected 18·4%, 15·6% and 46·9% infections whereas an in-house ELISA detected anti-trypanosomal antibodies in 42·1%, 43·8% and 65·6% infections in clinically ill buffaloes, cattle and horses, respectively. It can be concluded that T. evansi is prevalent in cattle, buffaloes and horses in the Eastern Region of India.