Opposite and synergistic physiological responses to water acidity and predator cues in spadefoot toad tadpoles

Organisms are exposed to multiple environmental factors simultaneously to which they often respond behaviorally, morphologically and/or physiologically. Amphibian larvae are quite plastic and efficiently adjust their phenotype and physiology to the reigning local conditions. Here we tested whether the combination of predator presence and low water pH induces alterations in the morphology and physiology of spadefoot toad tadpoles. We raised Pelobates cultripes tadpoles in the laboratory in water at either pH 4 or 7, and in the presence or absence of caged dragonfly nymphs, and determined their changes in shape through geometric morphometrics to assess whether predator recognition was impaired or not at low pH. We also measured levels of plasma corticosterone, activity of four antioxidant enzymes, as well as markers of oxidative damage and redox status. We found that tadpoles altered their body shape in response to predator cues even at low pH, indicating that predator recognition was not interfered by water acidity and developmental responses were robust even under abiotic stress. Water acidity was associated with increased corticosterone levels in tadpoles, whereas predator presence consistently reduced corticosterone levels. Predator presence was linked to reduced antioxidant enzyme activity, whereas the combination of both factors resulted in negative synergistic effects on lipid peroxidation and the antioxidant capacity of tadpoles. Here we show that tadpoles detect predators even at low pH but that the development of adaptive anti-predatory morphology can magnify physiological imbalances when other stressors co-occur. These results emphasize the need to understand how multiple environmental perturbations can affect animal homeostasis