Toxoplasma gondii is one of the most common foodborne protozoan parasite causing congenital infection, abortion, and stillbirth in humans and animals. The temperate and humid climate is one of the most important factors in the high prevalence of T. gondii. Sheep are among the important sources of meat production in Guilan province, Iran. Therefore, the consumption of raw and half-cooked meat is one of the major risk factors for T. gondii infection. Toxoplasmosis in patients with intact immune systems is usually asymptomatic; however, it but can be life-threatening in patients with a weak immune system (for example, patients with the human immunodeficiency viruses/acquired immunodeficiency syndrome or cancer and transplant recipients). Guilan is divided into three geographical regions of plains with a temperate climatic condition, hillsides with a semi-humid climate, and heights with cold mountainous climate. Climate situations play a role in the prevalence of toxoplasmosis. The present study aimed to investigate the seroprevalence of T. gondii infection among sheep in Guilan province, north of Iran. In the current cross-sectional study, a total of 400 sheep sera samples were tested for the determination of immunoglobulin G (IgG) antibody against T. gondii using the enzyme-linked immunosorbent assay. The samples were divided into different groups according to the geographical location and animal age. T. gondii antibody (i.e., IgG) was detected in 166 sheep (41.5%). The highest frequency of T. gondii infection (72.7%; n=56) was observed for the age group of > 4 years; the difference was statistically significant in this regard (P=0.0001) in comparison to that reported for other groups. In addition, the seroprevalence of T. gondii was significantly higher in the plains (53.9%) than that of the hillsides and heights (P=0.0001). Consequently, the seroprevalence of T. gondii infection in Guilan was high indicating a significant relationship with geographical location and animal age.