Molecular identification of badger-associated Babesia sp DNA in dogs: updated phylogeny of piroplasms infecting Caniformia

Background: Piroplasms are unicellular, tick-borne parasites. Among them, during the past decade, an increasing diversity of Babesia spp. has been reported from wild carnivores. On the other hand, despite the known contact of domestic and wild carnivores (e.g. during hunting), and a number of ixodid tick species they share, data on the infection of dogs with babesiae from other families of carnivores are rare. Methods: In this study blood samples were collected from 90 dogs and five road-killed badgers. Ticks were also removed from these animals. The DNA was extracted from all blood samples, and from 33 ticks of badgers, followed by molecular analysis for piroplasms with PCR and sequencing, as well as by phylogenetic comparison of detected genotypes with piroplasms infecting carnivores. Results: Eleven of 90 blood DNA extracts from dogs, and all five samples from badgers were PCR-positive for piroplasms. In addition to the presence of B. canis DNA in five dogs, sequencing identified the DNA of badger-associated Babesia sp. Meles-Hu1 in six dogs and in all five badgers. The DNA of Babesia sp. Meles-Hu1 occurred significantly more frequently in dogs often taken to forests (i.e. the preferred habitat of badgers in Hungary), than in dogs without this characteristic. Moreover, detection of DNA from this Babesia sp. was significantly associated with hunting dogs in comparison with dogs not used for hunting. Two PCR-positive dogs (in one of which the DNA of the badger-associated Babesia sp. was identified, whereas in the other the DNA of B. canis was present) showed clinical signs of babesiosis. Engorged specimens of both I. canisuga and I. hexagonus were collected from badgers with parasitaemia, but only I. canisuga contained the DNA of Babesia sp. Meles-Hu1. This means a significant association of the DNA from Babesia sp. Meles-Hu1 with I. canisuga. Phylogenetically, Babesia sp. Meles-Hu1 belonged to the B. microti group. Conclusions: This is the first detection of the DNA from a badger-associated Babesia sp. in dogs, one of which also showed relevant clinical signs. Based on the number of dogs with blood samples containing the DNA of Babesia sp. Meles-Hu1 in this study (i.e. exceeding the number of B. canis-positives), these findings should not be regarded as isolated cases. It is assumed that dogs, which are used for hunting or frequently visit forests, are more likely to be exposed to this piroplasm, probably as a consequence of infestation with I. canisuga from badgers or from the burrows of badgers. The above results suggest that Babesia sp. Meles-Hu1 should be added to the range of piroplasms, which are naturally capable of infecting hosts from different families of Caniformia.