Pertaining to reported high prevalence of brucellosis in Pakistan, district Layyah on account of having thick sheep population was selected for this study.  A total of 384 sheep blood samples were collected randomly from different selected private herds in the district, and tested through Modified Rose Bengal (mRB) test and Indirect Enzyme Linked Immunosorbent Assay (iELISA) for the serological analysis against the Brucella antibodies. Positive samples from these two tests were further subjected to Competitive Enzyme Linked Immunosorbent Assay (cELISA). The individual based seroprevalence of brucellosis in sheep was found to be 7.0% by mRB. Herd based prevalence was 42.5%. The highest (P<0.05) seroprevalence (16.8%) was observed in Tehsil Layyah as compared to Tehsils Karor (2.3%) and Choubara (4.5%). Non-significant differences were recorded between breeds, age and sex groups and also for the animals with or without history of abortion. In case of indirect and competitive ELISA, no sheep serum sample was found to be positive.

Key words: Brucellosis, Pakistan, Seroprevalence, Sheep, Southern Punjab