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An outbreak of buffalopox in domestic buffaloes, with high morbidity and significant production loss, was recorded in the Aurangabad district of Maharashtra State in India in November 2003. The disease was also associated with several cases of human infection, particularly in milkers working with the affected herds. Pox lesions were observed on the udder and teats of the majority of the affected animals, while a few animals exhibited lesions on the hindquarters, indicating possible generalised infection. A significant reduction in milk yield was recorded following the outbreak. Milkers developed pox-like lesions on the skin of the hands, forearms and forehead accompanied by fever for three days, axillary lymphadenopathy and general malaise. Investigation of the disease outbreak by virus isolation in Vero cell cultures and detection of viral nucleotide sequences by polymerase chain reaction (PCR) confirmed the aetiology of the disease.