Occurrence and molecular identification of haemotropic Mycoplasma species in grey wolves (Canis lupus) from southern Europe.

Although wild and domestic carnivores share some haemotropic Mycoplasma species, information about the circulation of this pathogen in grey wolves (Canis lupus) populations is still very limited. Thus, a geographically broad-based investigation was performed for determining the occurrence and diversity of Mycoplasma spp. in three different wolf populations from southern Europe. Between 2001 and 2023, spleen samples from 285 grey wolves from Spain (n = 129), Italy (n = 113), and Portugal (n = 43) were collected. The presence of haemotropic Mycoplasma was assessed targeting the 16S rRNA gene using two PCR assays in parallel; in addition, the 16S-23S rRNA intergenic spacer was analysed for further identification of the positive samples. The influence of the sampling country, sex, and age of the animals on the prevalence of Mycoplasma spp. was also assessed by a generalized linear model analysis. The percentage of positive wolves was 13.3 % (38/285), and the occurrence was significantly higher in Spain (20.9 %) than in Italy (8.0 %) and Portugal (4.7 %). Mycoplasma haemocanis (10.5 %) and Candidatus M. haematoparvum (2.1 %), were identified; in addition, an uncultured Mycoplasma sp. was also detected (0.7 %). Our results confirm the circulation of potentially zoonotic Mycoplasma in wolf populations from southern Europe. To our knowledge, this is the first report of Ca. M. haematoparvum in wolves from Italy and Portugal. In addition, a Mycoplasma sp., previously found in dogs, has been detected for the first time in wolves. Further studies are needed to fully molecularly characterise haemotropic Mycoplasma spp., which will serve as a basis for the study of its ecoepidemiology.