A study was conducted to identify the status of seroprevalence of leptospirosis in animal population by Microscopic Agglutination Test (antibody detection) covering different locations in Tamil Nadu, India. A total of 447 sera samples were collected with unknown disease status from different animal species viz., 108 canine, 102 bovine, 177 swine and 60 rodents. 47.2 (51/108) per cent of canine sera, 41.1 (42/102) per cent bovine sera, 15.8 (28/177) per cent swine sera and 40 (24/60) per cent rodent sera were positive for leptospirosis by Microscopic Agglutination Test. Most predominant serovars are Australis and Autumnalis in all species found in this study. Seroprevalence of new serovars other than vaccinal serovars indicates that these serovars may be included in vaccines preparation for prevention of leptospirosis outbreaks in animal and human population.