Temperature is considered an important factor that influences the life cycle of annual fishes, however the thermal preferendum of Austrolebias nigrofasciatus reproduction has not been defined. The hypothesis was that the fecundity of the species would respond differently within the temperature range analyzed, presenting an optimum thermal. Thus, this study investigated the effect of temperature on the fecundity of A. nigrofasciatus for the first time under laboratory conditions. Pairs of A. nigrofasciatus were placed in aquaria with temperatures of 17, 21 or 25°C, with four pairs (replicates) for each treatment, for five weeks. Contrary to expected, there were no significant differences in fecundity parameters among the tested temperatures, but the weekly fecundity at 17 and 21°C was more homogeneous than at 25°C. During the experimental period, the temperature did not affect male growth; however, at 25°C there was a negative effect on female body growth. The condition factor was also influenced, suggesting a tendency towards a negative energy balance with increasing temperature. The results indicate that temperatures between 17 and 21°C are suitable for broodstock maintenance.