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Background Theileria equi and Babesia caballi cause equine piroplasmosis (EP), one of the most important tick‐borne diseases of horses due to its high negative impact to the equine industry. Although infections with these parasites have been reported for decades in Spain, epidemiological studies have only been carried out in certain regions. Objectives To determine the (sero)prevalence of these parasites in asymptomatic horses nationwide in Spain and to identify potential individual and environmental factors associated with seropositivity to EP. Study design Sample size was calculated according to the horses registered in Spain in 2013 and by autonomous community using a random stratified sampling. A questionnaire was used to collect data on factors associated with EP seropositivity. Methods Serological (cELISA and complement fixation test) and molecular (real‐time PCR) analyses were carried out in 740 horses. Risk factors were identified computing two independent logistic regression models with the collated data. Results Antibodies against EP were detected in 42.9% (95% CI 39.4‐46.5) of horses, whereas 30.3% (95% CI 27.0‐33.6) were EP positive by PCR. A substantial strength of agreement (k = 0.721) was estimated between serological tests. Exposure to T. equi was significantly higher than to B. caballi and the highest (sero)prevalence was detected in the northern communities. Increasing horse age, presence of ticks and contact with cows were factors related to EP seropositivity in the horses, whereas tetanus vaccination and fairs attendance were associated with lower seropositivity. Conclusions Almost half of the horses residing in Spain had antibodies against EP or circulating parasitaemia. Appropriate prevention measures and implementation of a EP surveillance programme should be considered in order to reduce and control the infection.

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