Phyllostomidae, the most diverse family of Neotropical bats, encompass 230 species with varied dietary habits and food acquisition methods. Their feeding niche diversification has shaped skull and wing morphologies through natural selection, reflecting food processing and flight strategies. Yet, evolution of bat hindlimbs, especially in phyllostomids, remains little understood. Previous studies highlighted the femur's morphology as a key to understanding the evolution of quadrupedalism in yangochiropteran bats, including the adept walking observed in vampire bats (Desmodontinae). Here, we aimed to describe the femoral morphological variation in Phyllostomidae, correlating this with body size and assessing the effects of phylogenetic history, dietary habits, and hindlimb usage. Analyzing 15 femoral traits from 45 species across 9 subfamilies through phylogenetically informed methods, we discovered a significant phylogenetic structure in femoral morphology. Allometric analysis indicated that body mass accounts for about 85% of the variance in phyllostomid femoral size and about 11% in femoral shape. Relatively smaller femurs showed to be typical in Stenodermatinae, Lonchophyllinae, and Glossophaginae, in contrast to the larger femurs of Phyllostominae, Desmodontinae, Micronycterinae, and Lonchorrhininae. Furthermore, extensive femur shape variation was detected, with the most distinct morphologies in vampire bats, followed by frugivorous species. Adaptive evolutionary models related to diet more effectively explained variations in femoral relative size and shape than stochastic models. Contrary to the conventional belief of limited functional demand on bat femurs, our findings suggest that femoral morphology is significantly influenced by functional demands associated with diet and food capture, in addition to being partially structured by body size and shared evolutionary history.
