Toxoplasmosis is one of the most common meat‐borne parasitic infections worldwide. Consumption of raw or undercooked meat which contains Toxoplasma gondii tissue cysts is an important route of human infection. In this study, we investigate the serological and molecular prevalence of T. gondii infection in sheep and goat samples in Kashan, Iran, from 2015 to 2016. Serological (IgG antibody) and molecular detections were performed by the enzyme‐linked immunosorbent assay and polymerase chain reaction on the sera and heart samples of sheep (n = 90) and goats (n = 90), respectively. T. gondii‐IgG antibody was detected in 12.2% of sheep and 4.4% of goat samples. The parasite's DNA was detected in 17.8 and 8.9% of sheep and goat samples, respectively.Practical applicationsThe results of this study emphasize on the role of the sheep and goat as reservoirs of T. gondii infection. Hence, consumption of adequately cooked meat should be considered for prevention of T. gondii infection in humans.