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Diphylla ecaudata is a hematophagous bat endemic of South America, with food preference for bird blood. Given the lack of information about the reproductive activity of this species, this study aimed to describe the testicular morphology and histomorphometry of D. ecaudata in order to understand its reproductive biology, specially spermatogenesis. The animals were collected in Lajes city, Rio Grande do Norte, Brazil. Following euthanasia, the testes were histologically processed for morphological, morphometric, ultrastructural and immunohistochemical analyses. Their average body weight was 24.64g, with a gonadosomatic index of 0.49%, tubulesomatic index of 0.47%, and a total of 32.20m of seminiferous tubules per gram of testis. The pre-meiotic, meiotic, and post-meiotic phases accounted for 56.20%, 9.30%, and 34.50% of the seminiferous epithelium cycle, respectively. The ultrastructure of spermiogenesis was similar to that described in other mammals and the perforatorium was not observed in the sperm. Androgen receptors were detected in Sertoli cell nuclei and Leydig cell cytoplasm, while aromatase enzyme was detected only in Sertoli cell nuclei. FGF2 and BCL-2 activities were detected in the cytoplasm of zygotene and pachytene primary spermatocytes, as well as round and elongated spermatids. D. ecaudata showed testicular pattern similar to other mammals and characteristics common to other bat species. This species stood out for its high efficiency of Sertoli cells, which presented high capacity to support germ cells, besides the highest sperm production rates among those already recorded. This study is the first step towards the knowledge of D. ecaudata reproduction and the first description of its spermatogenesis. [ABSTRACT FROM AUTHOR]