Reintroduction of endangered fishes to historic habitat has been used as a recovery tool; however, these fish may face competition from other fishes that established in their native habitat since extirpation. This study investigated the physiological response of tidewater goby, Eucyclogobius newberryi, an endangered California fish, when competing for food with threespine stickleback, Gasterosteus aculeatus, a native species, and rainwater killifish, Lucania parva, a non-native species. Survival, growth and physiological indicators of stress (i.e. cortisol, glucose and lactate concentrations) were assessed for juvenile fish held for 28 days in two food-limited conditions. When fed a 75% ration, survival of E. newberryi was significantly lower when held with G. aculeatus. In all fish assemblages, weight and relative condition decreased then stabilized over the 28 day experiment, while length remained unchanged. Whole-body cortisol in E. newberryi was not affected by fish assemblage; however, glucose and lactate concentrations were significantly higher with conspecifics than with other fish assemblages. When fed a 50% ration, survival of E. newberryi decreased during the second half of the experiment, while weight and relative condition decreased and length remained unchanged in all three fish assemblages. Cortisol concentrations were significantly higher for all fish assemblages compared with concentrations at the start of the experiment, whereas glucose and lactate concentrations were depressed relative to concentrations at the start of the experiment, with the magnitude of decrease dependent on the species assemblage. Our findings indicate that E. newberryi exhibited reduced growth and an elevated generalized stress response during low food availability. In response to reduced food availability, competition with G. aculeatus had the greatest physiological effect on E. newberryi, with minimal effects from the non-native L. parva. This study presents the first reported cortisol, glucose and lactate concentrations in response to chronic stress for E. newberryi.