The low molecular weight and high cellular permeability of amides make them suitable for use as penetrative cryoprotectants for sperm cells.This study aims to evaluate the effect of dimethylformamide (DMF) and dimethylacetamide (DMA) on sperm cryopreservation of Curimba (Prochilodus lineatus).Semen samples were diluted in media containing cryoprotectants [DMF, DMA and dimethyl sulfoxide (DMSO)]. Parameters of motility, membrane integrity, DNA integrity, mitochondrial functionality, viability and fertility were assessed upon thawing.As compared to the 10% DMSO, DMA at 5% and DMF at 2% obtained the best results for the integrity of membrane, DNA and mitochondria; the motility parameters were best in the 2% and 5% DMF treatments. The best fertilization rates were demonstrated in 2%, 5%, and 8% DMF treatment groups.DMF at 2%, 5%, and 8% provided the best results for both in vitro and in vivo assessments, and can efficiently cryopreserve semen of Prochilodus lineatus.