Genetic diversity of equine piroplasms in Greece with a note on speciation within Theileria genotypes (T. equi and T. equi-like)

Equine piroplasms in Greece were studied using the reverse line blot hybridization (RLB) assay. Three genotypes consisting of two Theileria (T. equi and T. equi-like) and one Babesia (B. caballi-like) were identified. Of 787 samples tested, 371 (47.14%) hybridised to catchall probe (probe specifically designed to capture any piroplasm species present in a sample), 346 (43.96%) to T. equi probe, 364 (46.25%) to T. equi-like probe, 0 (0%) to B. caballi probe and 3 (0.38%) to B. caballi-like probe. Seven samples gave faint signals with the catchall probe only, indicating the presence of known or unknown piroplasm species, or a novel genotype or a known genotype occurring at a very low level of parasitemia. A partial sequence (509 bp) of the V4 region of the 18S rRNA gene of a T. equi-like isolate showed only 99% similarity with the reference T. equi-like isolates from Northern Spain from which the detecting probe used in the present study was designed but showed 100% similarity with the T. equi-like variants from Southern Spain. This indicated a noticeable degree of polymorphism within the population of T. equi-like. No unusual parasites previously reported in horses, such as B. canis canis and B. bovis were detected in this study. The values of the bioclimatic variables were very similar between the geographic locations for T. equi and T. equi-like genotypes, suggesting the two are not yet different species as hypothesized by some authors but are possibly undergoing a speciation process within Theileria genotypes. Both T. equi and T. equi-like were found in predominantly forest type land cover.