Abstract Cladocerans are able to produce resting eggs inside a protective resistant capsule, the ephippium, that difficults the visualization of the resting eggs, because of the dark pigmentation. Therefore, before hatching experiments, methods to verify viable resting eggs in ephippia must be considered. This study aimed to evaluate the number of eggs per ephippium of Daphnia from two tropical aquatic ecosystems and the efficiency of some methods for decapsulating resting eggs. To evaluate the influence of methods on hatching rates, three different conditions were tested: immersion in sodium hypochlorite, manually decapsulated resting eggs and intact ephippia. The immersion in hypochlorite solution could evaluate differences in numbers of resting eggs per ephippium between the ecosystems studied. The exposure to sodium hypochlorite at a concentration of 2% for 20 minutes was the most efficient method for visual evaluation and isolation of the resting eggs. Hatching rate experiments with resting eggs not isolated from ephippia were underestimated (11.1 ± 5.0%), showing the need of methods to quantify and isolate viable eggs. There were no differences between the hatching rate of resting eggs submitted to hypochlorite solution (47.2 ± 7.34%) and manually decapsulated (53.7 ± 13.24%). However, the immersion in hypochlorite was a more efficient technique, faster and not requiring manual ability.