Abstract: Bats and their ectoparasites play a crucial role in understanding the ecology and transmission of vector-borne pathogens, yet these dynamics remain poorly studied in Portugal. This study aimed to investigate the molecular occurrence of vector-borne bacteria (Anaplasma spp., Bartonella spp., Ehrlichia spp., and Rickettsia spp.) and protozoa (Babesia spp. and Theileria spp.) in ectoparasites of cave-dwelling bats. Bats were sampled from two caves in Portugal, and their ectoparasites included wing mites (Spinturnix myoti), ticks (Ixodes simplex), and bat flies (Penicillidia conspicua and Nycteribia schmidlii). Molecular analyses revealed the presence of Bartonella spp. in S. myoti and N. schmidlii. Phylogenetic inference based on the gltA gene positioned the detected genotypes close to those previously reported in bats and Nycteribiidae flies in Europe, Asia, and Africa. Notably, no DNA from Anaplasmataceae, Rickettsia spp., or piroplasmids was detected. The prevalence of S. myoti was high, with all examined bats being infested, showing notable differences in ectoparasite diversity concerning sex and cave-specific location. These findings suggest that host behavior, environmental conditions, and ectoparasite lifecycles play critical roles in shaping pathogen transmission dynamics. This study advances the understanding of bat ectoparasite–pathogen interactions in a region with limited data and highlights the need for continued research to assess the zoonotic potential and ecological impacts of the Bartonella genotypes detected herein.
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