Abstract
Hemotropic mycoplasmas (hemoplasmas) are uncultivable, cell wall-less bacteria that parasitizeon the surface of red blood cells of mammals, potentially causing anemia and other systemic signs. While widely distributed among domestic and wild animals, their occurrence in equids remains poorly understood, and no species has been identified as host-specific to horses or donkeys. This study presents the first systematic survey of hemoplasmas in equids from southeastern Europe and only the second molecularly confirmed case in horses in Europe. A total of 843 equids (817 horses and 26 donkeys) from different regions of Croatia, representing various ages, uses, and husbandry systems, were screened for hemoplasmas by PCR targeting the 16S rRNA gene. Only one horse tested positive, identified as Mycoplasma wenyonii, a hemoplasma typically associated with cattle. The estimated prevalence was 0.12% (95% CI: 0.003-0.68%). No donkeys were infected. The extremely low prevalence observed here-the lowest reported in any study detecting hemoplasma-positive horses-supports the hypothesis that equids do not harbor host-specific hemoplasma species and may only sporadically acquire infections from other hosts via spillover. This finding underscores the apparent absence of persistent hemoplasma lineages adapted to equids and highlights the need for further research on their epidemiology, host specificity, and transmission dynamics.