Sertoli cells play an essential role in spermatogenesis, being determinant of male reproduction capability. In this study we determined and compared the Sertoli cell efficiency (SCE), i.e. the ratio of the number of round spermatids to the number of Sertoli cells, of three species of Neotropical chiropterans, Anoura geoffroyi, Artibeus lituratus and Myotis levis, and additionally we tested for correlations between SCE, the combined mass of the testes and epididymides (CMTE), and the body condition index (BCI), considering that both BCI and testis mass can influence gonadal function of males. For SCE determination, the number of round spermatids and Sertoli cells present in stage 1 of the cycle of the seminiferous epithelium were counted. The Sertoli cell efficiency (mean ± 1 SD) was 4 ± 2.4 for A. geoffroyi, 4 ± 1.0 for A. lituratus and 6 ± 2.0 for M. levis. No significant variation was observed in SCE among the species, but A. lituratus exhibited a statistically significant correlation between CMTE and BCI. Anoura geoffroyi and M. levis exhibited significant positive correlations between SCE and CMTE, but not between SCE and BCI. Previous studies have shown that, unlike A. lituratus, A. geoffroyi and M. levis are subject to strong mating pressures, leading us to hypothesize that these findings may be related to differential mating pressures between species.