The sample size was calculated by the statistical formula presented by Naing et al. (2006). The prevalence estimation was made based on the previous article that had reported the prevalence of T. gondii in sheep (serology: 32.6% and PCR: 8%) and goats (serology: 48% and PCR: 11.3%) in Iran (Bahrami et al. 2019). For the purpose of this study, a slaughterhouse in the southwestern part of Iran (Ahvaz, Khuzestan province) was visited eight times, once a week during autumn 2018, by the researchers to collect blood samples from the common carotid arteries of a total of 150 sheep and 150 goats using a 10-mL vacutainer tube immediately after bleeding. The blood samples were then centrifuged at 4000 rpm for 5 min, and sera samples were separated and stored at − 20 °C for later serologic evaluation. Immediately after blood sampling from the slaughtered animals, their carcasses were opened and samples of blood, heart, liver, and meat (femur musculature) tissues were taken from 150 sheep (75 males and 75 females) and 150 goats (75 males and 75 females) using sterile single-use surgical blades and a 2-g piece of each tissue sample was then transferred into sterile plastic bags. The animals used were from eleven different farms which were randomly selected from the same slaughterhouse for the purpose of this study.

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