Bovine paratuberculosis is a chronic disease of cattle and buffaloes, causing progressive weight loss, persistent diarrhea and finally death. Due to zoonotic nature of the disease, workers in slaughterhouses are at a high risk of infection. In this study, the status of paratuberculosis was assessed in a slaughterhouse located in district Faisalabad, Pakistan. A total of 455 blood samples were collected from slaughtered cattle and buffaloes at random and then tested by a commercially available ELISA kit for the detection of antibodies against Mycobacterium avium subsp. paratuberculosis (MAP). Data were analyzed using chi square for ELISA and odds ratio for finding association of species, gender, body weight, age and body condition score (BCS) with prevalence of antibodies against MAP. Seropositivity against MAP was significantly (P<0.05) detected. Male and female animals were equally susceptible to the disease (OR: 0.908, 95% CI= 0.6111-1.350). Age groups were not found associated with chances of being seropositive. Seropositivity was significantly (P<0.05) higher in animals under 300kg body weight. Chances of being seropositive were found more in animals belonging to BCS 1 (OR: 13.25, 95%CI=6.29-27.88) and BCS 3 (OR: 4.78, 95%CI=2.36-9.70) than BCS 4 and above. In conclusion, gender, body weight and body condition score of the animals are positively associated with the occurrence of the disease. These data will not only help in screening of animals but will also be instrumental in future for isolation of MAP to make vaccine against paratuberculosis using local isolates.