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Bacteria in the genus Borrelia are primarily spread by ticks and cause either Lyme borreliosis or relapsing fever. Substantial work has demonstrated the degree to which rodents and songbirds can contribute to the enzootic cycles and dispersal of these human diseases, but comparatively less attention has been paid to the role of wild bats, particularly in temperate regions. We here report human-relevant findings from a two-year, seasonal survey of migratory Mexican free-tailed bats ( Tadarida brasiliensis ) in Oklahoma, USA. We tested nearly 400 bats and identified Borrelia puertoricensis , a relapsing fever species that could infect humans. Importantly, this represents the first detection of Borrelia puertoricensis in bats and only the second detection in wild vertebrate hosts, expanding the known host range of this emerging tick-borne pathogen. Given the known migratory routes of Mexican free-tailed bats, our results have implications for the role that bats may play in tick-borne pathogen dispersal in North America.

Bats have been intensively sampled for viruses but remain mostly understudied for bacterial pathogens. However, bacterial pathogens can have significant impacts on both human health and bat morbidity and even mortality. Hemoplasmas and bartonellae are facultative intracellular bacteria of special interest in bats, given their high prevalence and substantial genetic diversity. Surveys have also supported plausible zoonotic transmission of these bacteria from bats to humans, including Candidatus Mycoplasma haematohominis and Candidatus Bartonella mayotimonensis. Greater characterization of these bacteria across global bat diversity (over 1,480 species) is therefore warranted to inform infection risks for both bats and humans, although little surveillance has thus far been conducted in North American bats. We here describe novel (and in some cases panmictic) hemoplasmas and bartonellae across three colonies of Mexican free-tailed bats and sympatric bat species. We find high genetic diversity and seasonality of these pathogens, including lineages closely related to human pathogens, such as Bartonella rochalimae .

Identifying the drivers of wildlife immunity is critical for assessing stressor impacts and zoonotic risks. However, such studies are limited by logistical challenges of wildlife research and lack of species-specific reagents. We adapt flow cytometry, typically confined to laboratory settings, to field settings to profile cellular immunity with small blood volumes and extended sample holding times. We apply these methods to analyze immune cell seasonality in migratory Mexican free-tailed bats ( ). We confirmed four antibodies recognizing CD3, CD79a, MHCII, and CD11b that were originally validated in Egyptian fruit bats ( ), allowing us to quantify T and B cells, macrophages, and neutrophils, respectively. Flow cytometry outperformed hematology in quantifying leukocyte profiles and revealed pronounced immune cell seasonality. Adaptive cells steadily increased between spring and fall migration. Neutrophils were most abundant during the reproductive period and decreased during migrations, whereas B cells were most abundant after reproduction and before fall migration; granulocytes as a whole, macrophages, and T cells had no seasonality. Females had more B cells than males but did not differ in other cells. Our findings lay the groundwork for applying flow cytometry to field studies of wildlife and provide important insights into the seasonality of bat immunity.

We detected in migratory Mexican free-tailed bats sampled in Oklahoma during 2022 and 2023, representing only the second detection of this relapsing fever species in wild vertebrates. Although prevalence was low (0.79%), our findings suggest migratory bats could contribute to dispersal of tick-borne pathogens in North America.

Bacterial pathogens remain poorly characterized in bats, especially in North America. We describe novel (and in some cases panmictic) hemoplasmas (10.5% positivity) and bartonellae (25.5% positivity) across three colonies of Mexican free-tailed bats ( ), a partially migratory species that can seasonally travel hundreds of kilometers. Molecular analyses identified three novel hemoplasma species most similar to another novel species in Neotropical molossid bats. We also detected novel hemoplasmas in sympatric cave myotis ( ) and pallid bats ( ), with sequences in the latter 96.5% related to Mycoplasma haemohominis. We identified nine genogroups, including those in cave myotis with 96.7% similarity to Bartonella mayotimonensis. We also detected in migratory Mexican free-tailed bats, representing the first report of this human pathogen in the Chiroptera. The seasonality and diversity of these bacteria observed here suggest that additional longitudinal, genomic, and immunological studies in bats are warranted.