This study was carried out to investigate the incidence of Toxoplasma gondii in 105 sheep and goat raw milk samples in Assiut, Egypt. Milk samples were tested by microscopic examination, Latex agglutination test and 50 samples were tested with PCR assay. The incidence of Toxoplasma gondii in sheep and goat milk samples was 10.48% (11/105) by microscopic examination, 39% (41/105) by Latex agglutination test and 16% (8/50) by PCR assay. High significant differences were recorded in incidence of Toxoplasma in raw sheep milk in 8.62% (5/58) by micropscopic examination, in 39.66% (23/58)) by latex agglutination test and detection of B1 gene of Toxoplasma gondii in 10.71% (3/28)by PCR assay. While,significance differences were found in incidence of Toxoplasma inraw goat milk in 12.77% (6/47)) by microscopical examination, in 38.30%(18/47) by Latex agglutination test in addition to detection of B1 gene of Toxoplasma gondii in 22.73% (5/22) by PCR assay. The results showed that sheep and goats were excreting Toxoplasma gondii DNA in their milk. The results of this study revealed that Latex agglutination test and PCR assay could become useful tools to diagnose the incidence of Toxoplasma gondii in sheep and goat milk samples. This study is the first report of direct detection of Toxoplasma gondii in sheep and goat milk samples in Assiut Governorate, Egypt.Control measures and public heath importance were also discussed in this study.