Abstract One hundred and twenty serum samples from ostriches (Struthio camelus) in a commercial farm in Egypt were tested for anti-Toxoplasma antibodies using ELISA test (IgM and IgG ) and Modified agglutination test (MAT). Five out of 120 birds (4.2%) were found positive to IgM enzyme immunoassay and 11 birds (9.2%) were IgG enzyme immunoassay positive. Modified agglutination test detected IgG in 15 birds (12.5%). PCR amplification of Toxoplasma gondii DNA in the blood of birds was recorded in 9 birds (7.5%). The Cases were categorized into acute and chronic cases depending on the result of blood-PCR and serological test where the positive cases for IgG were the most frequent (6.7%). The tissues of five dead birds tested by PCR assay showed the amplification of DNA fragment of T. gondii from the heart and brain of two birds, from only the brain of another two birds and from the heart, brain and leg muscles of one bird. The present study concluded that, the chronic cases are the most prevalent among ostriches population and the specific serological tests as MAT alone or with ELISA can be used as a highly sensitive screening test followed by PCR as a confirmatory test for diagnosis of toxoplasmosis and for detection of primary acute and chronic toxoplasmosis in ostriches. Key word: Toxoplasma, Ostriches, ELISA, MAT, PCR, Egypt.