<p><i>Toxoplasma gondii</i> is a zoonotic protozoan parasite with a wide host range, including cats, which are its definitive hosts. This study aimed to determine the prevalence of <i>T. gondii</i> infection among domestic and street cats in Golestan Province, Iran. A total of 94 cats, comprising 30 domestic and 64 street cats, were randomly selected and tested for <i>T. gondii</i> antibodies using ELISA. The overall prevalence of <i>T. gondii</i> was found to be 24.47% (23/94), with infection rates of 23.3% in domestic cats and 25% in street cats. Age distribution analysis revealed that 40.43% of the cats were 1–2 years old, 38.30% were 2–3 years old, and 21.27% were over 3 years old. Additionally, 20% of male cats (9/45) and 28.57% of female cats (14/49) tested positive for <i>T. gondii</i>. These findings indicate a significant presence of <i>T. gondii</i> in both domestic and street cats in Golestan Province, highlighting the need for effective control measures to prevent the transmission of the parasite to humans and other animals. Further research is required to explore the transmission dynamics of <i>T. gondii</i> and to develop strategies for the prevention and management of the disease in cat populations.</p>