In this study serum samples from 1067 horses residing in different provinces in Spain were analysed. The samples were received from August 2015 to August 2016, included 517 male horses, 316 mares and 234 samples from horses with unknown sex) and were tested using two commercial diagnostic kits: Babesia equi Antibody Test Kit and Babesia caballi Antibody Test Kit (VMRD, Pullman), in order to detect immunoglobulins type G against these parasites. For each sample, the inhibition percentage was calculated and only the samples with a percentage overo or equal to 40% were considered positive.