Toxoplasmosis is a significant public health concern, primarily associated with consuming raw or undercooked dairy products. However, there is limited knowledge concerning the seroprevalence of Toxoplasma gondii in dairy cattle slaughtered in high-throughput abattoirs intended for human consumption in South Africa. This study aimed to determine the seropositivity of toxoplasmosis in dairy cattle slaughtered at three high-throughput abattoirs. In 2023, 150 blood samples were randomly collected from dairy cattle at three high-throughput abattoirs in the Eastern Cape Province, South Africa. The seropositivity was significantly higher in abattoir B (East London), which had 30% positive dairy cattle. No significant difference was observed in abattoirs A (Port Elizabeth) and C (Queenstown). In animals younger than 3 years, 22% were infected with t. gondii. The Friesland breed showed more susceptibility than jersey and crossbreed breeds. The chi-square test was used to determine associations between the prevalence of toxoplasmosis, where low seropositivity resulted from the data, showing the disease to be less likely to occur. This research represents one of the few studies on the seropositivity of T. gondii in dairy cattle in South Africa. While the prevalence of infection was found to be low from the seropositivity results of the dairy animals, our findings underscore the potential public health implications associated with T. gondii in dairy cattle based on the low level of knowledge on the disease. The study results were found to be less significant; the null hypothesis was accepted. Recognizing the significance of T. gondii infection in dairy products is critical. This study contributes to our understanding of the prevalence of T. gondii in South African dairy cattle, highlighting the need for continued monitoring and control measures to ensure the safety of dairy products for consumers.