| Summary: | <i>Toxoplasma gondii</i> is a worldwide zoonotic parasite. According to the “One Health” approach, studies on toxoplasmosis are essential since it affects humans and domestic and wild animals. In the present study, antibodies to <i>T. gondii</i> were determined in serum samples from 263 wild birds located in five wildlife rehabilitation centres in mainland Portugal by using the modified agglutination test (MAT) with a cut-off titre of 20. An overall seroprevalence of 36.5% (95% confidence interval [CI]: 30.7–42.6) was observed. For the first time, antibodies to <i>T. gondii</i> were detected in some avian species, including pallid swift (<i>Apus pallidus</i>) (33.3%), black-backed gull (<i>Larus fuscus</i>) (39.3%), European turtle-dove (<i>Streptopelia turtur</i>) (100%), bee-eater (<i>Merops apiaster</i>) (50.0%), carrion crow (<i>Corvus corone</i>) (33.3%), and Egyptian vulture (<i>Neophron percnopterus</i>) (100%), which expands the list of intermediate hosts of <i>T. gondii</i>. A lower seroprevalence was found in juvenile birds (31.9%) compared to adults (48.7%) (<i>p</i> = 0.016). The central region of Portugal was considered a risk factor for <i>T. gondii</i> infection in wild birds (odds ratio: 3.61; 95% CI: 1.09–11.91). This pioneer study calls attention to the need for further studies, to provide a clearer understanding of <i>T. gondii</i> epidemiology in Portugal, because it reflects wide dispersion of <i>T. gondii</i> oocysts in the environment.
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