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Background: The parasites Toxoplasma gondii (T. gondii) and Neospora caninum (N. caninum) are globally distributed; they infect warm-blooded animals, including many avian species. The aim of this study was to evaluate the presence of these parasites in ostriches from central China. In total, 402 ostrich (Struthio camelus) samples (293 hearts, 77 brains, and 32 serum) from slaughterhouses of the Henan Province and Hebei Province were collected. The heart juice (n = 283) and serum samples (n = 32) were tested for antibodies to T. gondii using the modified agglutination test (MAT). Hematoxylin and eosin (H&E) staining, immunohistochemical (IHC) staining, and the polymerase chain reaction were used to examine the cysts and DNA of T. gondii and N. caninum parasites, respectively., Results: Antibodies to T. gondii were detected in 6.4% (20/315) (cut-off, 25). No cysts or DNA of T. gondii or N. caninum were observed in any of the 293 hearts and 77 brains., Conclusion: The results showed a low prevalence of T. gondii antibody in ostriches, compared to that in the other animals. N. caninum occurs at low to negligible frequencies in ostriches from China. This is the first report on screening ostriches in China for T. gondii antibodies.