Equine piroplasmosis in different geographical areas in France: Prevalence heterogeneity of asymptomatic carriers and low genetic diversity of Theileria equi and Babesia caballi

Equine piroplasmosis is a worldwide tick-borne disease caused by the parasites Theileria equi sensu lato and Babesia caballi, with significant economic and sanitary consequences. These two parasites are genetically variable, with a potential impact on diagnostic accuracy. Our study aimed to evaluate the frequency of asymptomatic carriers of these parasites in France and describe the circulating genotypes. We developed a species-specific nested PCR protocol targeting the 18S small sub-unit (SSU) rRNA gene and used it on blood samples collected from 566 asymptomatic horses across four National Veterinary Schools. The carrier frequency varied from 18.7 % around Paris (central-north) to 56.1 % around Lyon (southeast), with an overall prevalence of 38.3 %. Theileria equi carriers were ten times more frequent (91.7 %; 209/228 isolates) compared to B. caballi carriers (8.3 %; 19/228 isolates). Notably, T. equi carrier frequency was significantly lower in the northern region (Ile de France) compared to the southeastern regions. A positive relationship was observed between the frequencies of asymptomatic carriers and the frequency of previous acute piroplasmosis reported from the owner across all four geographic areas. Neither horse gender nor age showed a significant effect on the frequency of asymptomatic carriers. In some areas, a substantial proportion of horses (22.2 % to 37.5 %) carried T. equi before the age of three years, indicating high infection pressure. Genotyping of 201 T. equi isolates revealed a predominance of genotype E (98 %) and few isolates belonging to genotype A (2 %). Notably, two of the four T. equi genotype A isolates were detected in horses originating from Spain. All 19 B. caballi isolates belonged to the genotype A. The discussion section explores the link between these results, the tick distribution and abundance, and the frequency of detection of T. equi and B. caballi in febrile cases attributed to piroplasmosis.