The Effect of Venipuncture Site on Hematology of Bats: Implications for Comparative Analyses

Wildlife health comparisons within and across populations and species are essential for population assessment and surveillance of emerging infectious diseases. Due to low costs and high informational yield, hematology is commonly used in the fields of ecoimmunology and disease ecology, yet consistency and proper reporting of methods within these fields are lacking. Previous investigations on various wildlife taxa have revealed noteworthy impacts of the vein used for blood collection on hematology measures. However, the impacts of venipuncture site on bats, a taxon of increasing interest in ecoimmunology and disease ecology, have not yet been tested. Here, we use a long-term study system in western Oklahoma to test the effect of venipuncture site on hematology parameters of the Mexican free-tailed bat (Tadarida brasiliensis) and cave myotis (Myotis velifer), two abundant and representative bat species from the families Molossidae and Vespertilionidae. Between September 2023 and October 2024, we collected paired peripheral blood from both the propatagial and intrafemoral veins in 25 individuals per species. We then quantified total red and white blood cells, reticulocyte counts, and leukocyte differentials and used generalized linear mixed models to compare parameters among venipuncture sites within and between bat species. Overall, venipuncture site had no effect on any hematology parameters; however, we revealed small differences in neutrophil and lymphocyte proportions between veins among the species. By contrast, we detected significant species-level differences in most cell measurements, which we propose could be explained by life-history strategy and phylogenetic differences. We encourage continued testing of additional venipuncture sites, and of the same venipuncture sites on different species, on hematology and other health metrics used in ecoimmunology and disease ecology. Lastly, we emphasize the importance of thorough method reporting in publications to enable transparent comparisons and accounting for even small sampling-based artifacts. All future efforts are especially important for bats to improve conservation monitoring, ecosystem services estimations, and their association with emerging infectious diseases.