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The sperm ultrastructure of Epipedobates trivittatus and E. hahneli is described. The spermatological characteristics are identical in both species, except for the extension of the acrosomal vesicle. The structure of the acrosomal complex is shared with species of the Bufonoidea neobatrachian lineage, which differs from the condition observed in species included in the ranoid lineage. The phylogenetic significance of the mitochondrial collar, also seen in Eubufonoidea groups, is still unclear. Also the significance of the alternative mitochondrial arrangement seen here is unclear. The perpendicular arrangement of the centrioles is also shared with the Bufonoidea and is considered a synapomorphic condition. An expanded undulating membrane and the absence of a juxtaxonemal fibre appear to be apomorphic characteristics of dendrobatid species, as also observed (homoplastically) for species of Litoria and Cyclorana. However, the notable expansion of the undulating membrane appears to be an autapomorphy of Dendrobatidae frogs. The ultrastructural data presented here do not support the proposed placement of E. trivittatus in a separate genus.