The aim of this study was to e valuate the performance of dimethylacetamide (DMA) as cryoprotectant at three concentrations: 8% (0.85 M), 10% (1.07 M) and 12% (1.29 M), combined with glucose at 6% (0.33 M) and egg yolk at 12%, to cryopreserve semen of bocachico Prochllodus magdalenae. The semen was diluted at 1:4 ratio in the cryoprotective solution, packed in 2.5 mL straws and frozen in liquid nitrogen vapor (LN2) for 30 minutes, then it was stored in a container with LN2 of 34 L. The concentration, total motility, progressive motility and sperm speed in both fresh (control) and cryopreserved-thawed semen were evaluated using Sperm Class Analyzer (SCA). Straws were thawed in serological bath at 60 degrees C for 45 seconds. Fertility and hatching were evaluated inseminating 2 g of eggs (1,540 eggs/g) at a rate of 320,000 spz/egg. Fresh semen recorded the highest values of total motility, total progressive, curvilinear speed and linear speed with regard to cryopreserved-thawed semen (P < 0.05); whereas cryopreserved-thawed semen with DMA 8% and 10% recorded the higher values for these variables, without significant differences between them (P > 0.05). DMA 12% reported the lowest values of all the variables evaluating semen quality. Fresh semen obtained values of 74.0 +/- 5.3% and 62.3 +/- 3.1% for fertility and hatching, respectively, while cryopreserved-thawed semen with DMA at 8% produced adequate fertility (60.4 +/- 8.4%) and hatching (50.1 +/- 9.8%) values. The results of this study suggest that the cryoprotectant solution composed of DMA at 8%, glucose at 6% and egg yolk at 12% is a viable alternative for the cryopreservation of semen of bocachico.