The Toxoplasma gondii (Nicole and Manceaux, 1908) (Apicomplexa: Coccidia) is an obligate intracellular protozoan parasite, causing toxoplasmosis in humans and cattle worldwide. In the current research, seroprevalence and risk factors of toxoplasmosis in the cows (cattle), Bos indicus Linnaeus 1758 (Mammalia: Eutheria) of Dir Upper (DU), Khyber Pakhtunkhwa (KP), Pakistan were determined. Bos indicus (nt=250) including males (nm=86) and females (nf=164) were tested for the presence of antibodies to T. gondii using Latex Agglutination Test kit ® . Agglutination at 1:16 dilution was considered positive. A 14% overall prevalence was recorded. The antibody titers to T. gondii positive sera were 10%, 2%, 1% and 0% by dilution of 1:16, 1:32, 1:64 and 1:128, respectively. However, males were found more infected (16%) compared to females (12%). Moreover, the highest rate (16%) of infection was recorded in age 1-4 years, while the lowest (7%) was in 5-8 years. Further, no significant difference was found between toxoplasmosis and sex/age. Furthermore, infection was the highest (25%) in B. indicus with 5 pregnancies and no abortion was observed in infected B. indicus. In addition, presence of cats, Felis catus Linnaeus, in households, source of drinking water and type of management of B. indicus were risks for the same. In conclusion, toxoplasmosis is prevalent in B. indicus in DU. Prompt control measures and public awareness is recommended to lower the risk of infection with T. gondii.