Toxoplasmosis is very important zoonotic disease in the world cause by an obligate intracellular protozoan parasite called Toxoplasma gondii can infect human and all warm-blood animals, beef consider from most important source for infection with T. gondii and there is no really data and study about the rate of the infection in beef in Al-Diwaniyah province, so for this reason the aim of this work was designed. A total of 300 samples which collected from heart, tongue, muscles, of 100 slaughtered beef of local and imported cattle, throughout the period from September 2017 to May 2018, initially examined microscopically for searching on T. gondii bradyzoites then all suspected samples was subjected to conventional PCR technique through B1gene amplification to confirm the infection, in addition to analyzed the recorded data for each sample to determine the effect of some factors on prevalence of infection like organ, season and animal age. Out of 300 tested samples only 53 were confirm positive T. gondii DNA. The infection in local beef was higher (22%) than in imported (13.5%), while there is no difference in infection among different examined organs. Regarding to effect of some factors, the autumn season recorded highest rate of infection with significant differences rather than others seasons in both local and imported beef, whereas, age appears with no effect on infection. The local cattle meat is riskier than the imported due to the higher rate of infection with T. gondii, and the animal age cannot affect on the infection rate, in comparing with the season which play role in this rate.