, a newly identified member of the complex, has been isolated predominantly from hoofstock in eastern Africa and the Arabian Peninsula, and sporadically in cattle (), rhesus monkeys (), humans, and a greater one-horned rhinoceros () in South Asia. In rhinoceros, tuberculosis typically presents as a chronic progressive respiratory disease. The report describes the postmortem diagnosis of tuberculosis caused by in a greater one-horned rhinoceros with hind limb paresis due to neural granulomatosis. Serologic assays for detection of antibodies to complex proteins before culture results allowed for appropriate herd management protocols to be initiated. genus-specific polymerase chain reaction assays with direct sequencing allowed timely confirmation of the serologic results. This is the first isolation of in the western hemisphere, showing the need for mycobacterial testing of rhinoceros before international shipments and the urgency for validated antemortem complex screening assays in rhinoceros species.