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The prevalence of piroplasm (order Piroplasmida) infection was assessed in blood and bone marrow samples from 91 red foxes (Vulpes vulpes) from northern, central and southern Portugal by means of molecular methods. PCR for the 18S rRNA gene of Babesia spp. followed by sequencing revealed 63 foxes positive for the Babesia microti-like piroplasm (syn. Theileria annae) (69.2%; 95% confidence interval [CI]: 58.7-78.5%) and one fox positive for Babesia canis (1.1%; 95% CI: 0.0-6.0%). Positivity to the B. microti-like piroplasm or B. canis in 43 blood samples (83.7%) was significantly higher (p<0.001) than in 43 paired bone marrow samples (20.9%). There were no statistically significant differences in the prevalence of infection between genders (p=0.219) or age groups (<2 years vs. ≥ 2 years) (p=1.0). This is the first report of the B. microti-like piroplasm in foxes from Portugal as well as the first report on detection by PCR and genotyping of B. canis in a red fox worldwide. A natural cycle of the B. microti-like piroplasm is suggested in red fox populations based on the high prevalence of the protozoan. Red foxes might be a reservoir of the B. microti-like piroplasm and a source of infection to dogs.

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We report an investigation of antibodies to Toxoplasma gondii in 52 wild birds and 20 wild mammals from northern and central areas of Portugal by using the modified agglutination test. The birds comprised 26 common buzzards (Buteo buteo), five tawny owls (Strix aluco), four white storks (Ceconia ceconia), three Eurasian eagle owls (Bubo bubo), three northern goshawks (Accipiter gentilis), two booted eagles (Hieraaetus pennatus), two common barn owls (Tyto alba), two Eurasian sparrowhawks (Accipiter nisus), two short-toed eagles (Circaetus gallicus), one black kite (Milvus migrans), one Griffin vulture (Gyps fulvus), and one peregrine falcon (Falco peregrinus). The mammals were eight wild boars (Sus scrofa), six red foxes (Vulpes vulpes), two common genets (Genetta genetta), two European badgers (Meles meles), one European roe deer (Capreolus capreolus), and one Iberian wolf (Canis lupus signatus). Fifty percent of the wild birds and 90% of the wild mammals were seropositive; the overall seroprevalence of infection was 61.1%. When comparing the prevalence of antibodies in birds and mammals from northern Portugal, a significant difference was found, but the same was not true for birds and mammals from central Portugal. Seroprevalence levels were 30.0% in juvenile and 62.5% in adult birds (p=0.046), 0.0% in juvenile and 94.7% in adult mammals (p=0.100), 80.0% in female and 66.7% in male birds (p=1.000), and 81.8% in female and 100% in male mammals (p=0.479). This is the first study performed on T. gondii in birds of prey, white storks, and wild carnivores in Portugal.