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Aim: The present study was undertaken to know the seroprevalence of brucellosis in cattle and buffaloes. The work was carried out during the period 2011 through 2013 at Centre for Animal Diseases, Research and Diagnostics, IVRI, Izatnagar, Uttar Pradesh, India. Materials and Methods: A total of 1005 sera samples were collected and tested for bovine brucellosis using ELISA Kit; IDEXX, CHEKIT, Brucellose serum, Brucella abortus Antibody Test Kit. Results: A total of 1005 serum samples were collected from Karnataka, Uttar Pradesh and Uttarakhand during the period of 2011 to 2013 and were screened for bovine brucellosis using i-ELISA (indirect enzyme linked immunosorbent assay). The prevalence of bovine brucellosis was compared between organized and unorganized farms in order to find the epidemiology of the disease among the animal population. Sera from 5 organized farms in Karnataka were collected for seroprevalence studies. Out of 417 animals, 191 (45.80%) animals were found positive by i-ELISA. A total of 361 serum samples were collected from 5 unorganized farms or villages, of which 82 (22.71%) were positive. From Uttar Pradesh, bovine serum samples were collected from 3 organized farms. Out of 192 animals, 43 (22.39%) animals were found positive for brucellosis. Similarly, sera collected from a single organized farm from Uttarakhand, showed 3 (8.57%) positivity among 35 animals. On the whole, 319 (31.74%) animals were found positive for brucellosis among the 3 states taken for study, which includes 138 (27.21%) cattle and 181 (36.34%) buffaloes. Conclusion: It has been found that Brucella infections are widely prevalent in organized and unorganized dairy farms in investigated states of India. [Vet World 2013; 6(8.000): 550-553]