Toxoplasma gondii is a single-celled parasite that has a wide range of hosts including humans. A cross-sectional survey was conducted to investigate T. gondii seroprevalence and associated risk factors in small ruminants, pigs, poultry and cats in the Oliver Reginald Tambo District in the Eastern Cape in South Africa between June 2016 and October 2016. Household-level and animal-level data were collected using a close-ended questionnaire. One sample of each present species was collected in each household. The Toxoreagent ®, Mast Group, United Kingdom, latex agglutination test, was used for T. gondii antibody detection. Positive samples had agglutination patterns at dilutions of 1:64 or greater, except for chickens, whose cut off titre was 1:32. A household was classified as T. gondii seropositive if at least one species tested positive. The study revealed that 78 out of 121 sheep (64.46%), 69 out of 128 goats (53.91%), 36 out of 106 pigs (33.96%), 35 out of 109 cats (32.11%) and 46 out of 137 chickens (33.58%) were seropositive for the parasite. Seropositivity was assessed for association with potential risk factors. Age, location, climate, animal production system, rodent control, cat-feed access and cat faecal disposal were found to be significantly associated with seropositivity using the Chi-Squared test or odds ratio confirmed by the Fisher's exact test. The relatively high seroprevalence of T. gondii detected in this study suggests that the infection T. gondii poses a substantial public health risk through the consumption of infected raw or undercooked meat infected with T.gondii cysts as well as contact with cat faeces infected with T. gondii oocysts.