Chronic Cu toxicity was evaluated in the euryhaline copepod Acartia tonsa. Male and female copepods were exposed (6 d) separately to different combinations of Cu concentration and water salinity (5, 15, and 30 ppt) using different routes of exposure (waterborne, waterborne plus dietborne, and dietborne). After exposure, groups of one male and three female copepods were allowed to reproduce for 24 h. In control copepods, egg production augmented with increasing water salinity. However, egg hatching rate did not change. Copper exposure reduced egg production and hatching rate in all water salinities tested, but the reproductive response was dependent on the route of Cu exposure. Median effective concentration (EC50) values for egg production after waterborne exposure were 9.9, 36.8, and 48.8 µg/L dissolved Cu at water salinities of 5, 15, and 30 ppt, respectively. For waterborne plus dietborne exposure, they were significantly higher (40.1, 63.7, and 109.9 µg /L, respectively). After dietborne exposure, approximately 40% decrease in egg production was observed, independently of Cu concentration and water salinity tested. At water salinities of 5 and 30 ppt, egg hatching rate reduced after waterborne exposure, together or not with the dietborne exposure. At water salinity of 15 ppt, Cu toxicity was only observed after dietborne exposure. Data indicate that egg production is a more reliable reproductive endpoint to measure chronic Cu toxicity in copepods than egg hatching rate in a wide range of water salinities. They also suggest that both water salinity and route of Cu exposure should be taken into account in the development of a chronic biotic ligand model version for estuarine and marine environments.