Infections in wild carnivores by Babesia sp., Anaplasma sp., and Mycoplasma spp.

Abstract

The aim of this research was to investigate the prevalence and genetic diversity of Babesia sp., Anaplasma sp., and Mycoplasma spp. infections in wild carnivores, contributing to a better understanding of their role as reservoirs for zoonotic pathogens and the dynamics of vector-borne disease transmission in wildlife populations. Seventy spleen samples from wolves (Canis lupus) and four samples from golden jackals (Canis aureus) were collected between 2018 and 2024 in Lithuania and stored frozen until analysis. Genomic DNA was extracted from the spleen tissues of wild carnivores using the Genomic DNA Purification Kit (Thermo Fisher Scientific, Vilnius, Lithuania), following the manufacturer’s recommendations. The presence of protozoan and bacterial pathogens was screened using conventional and nested PCR techniques, with positive and negative controls included in each PCR run. PCR products were visualized by electrophoresis on a 1.5% agarose gel. Positive PCR products were extracted from the gel and purified using the GeneJET™ Gel Extraction Kit (Thermo Fisher Scientific, Vilnius, Lithuania). Partial sequences of Babesia sp., Anaplasma sp., and Mycoplasma spp. were analyzed using the MegaXI software package and aligned with previously uploaded sequences in GenBank via BLASTn. Phylogenetic trees were constructed using the maximum-likelihood method with the Tamura-Nei model. The study showed that Babesia canis was the most widespread pathogen, detected in 38% of the tested individuals. Anaplasma phagocytophilum was identified in 12% of the samples, while Mycoplasma spp. infections were found in 7% of the tested carnivores. These results highlight the significant role of wild carnivores as potential reservoirs for zoonotic pathogens, with important implications for wildlife conservation and public health strategies.