The impact of elevated temperature and CO(2) on growth, physiological and immune responses of Polypedates cruciger (common hourglass tree frog)

BACKGROUND: Amphibians are one of the most susceptible groups to climate change as their development occurs in aquatic environments or in microhabitats with high humidity. Accordingly, our primary objective was to investigate the chronic physiological responses seen in early larval to adult stages of Polypedates cruciger (Common hourglass tree frog) to future climate change based on continuous exposure to elevated temperature and elevated CO(2) -induced low water pH. Free-swimming and free-feeding tadpoles were observed until metamorphosis under four experimental treatments; two elevated temperatures, one elevated CO(2) (reduced pH) and a control maintained at ambient temperature (29 °C ± 1 °C) and CO(2) (pH = 7). Elevated temperature treatments were maintained at 32 °C ± 0.5 °C and 34 °C ± 0.5 °C to represent respectively, the future climate scenarios RCP2.6 (Representative Concentration Pathway 2.6, the ‘base-case’ scenario) and RCP8.5 (‘business-as-usual’ scenario) according to the 5(th) Assessment Report of the IPCC. Elevated CO(2) treatment was maintained within the pH range of 5.5–5.6 representing the range expected between RCP8.5 and RCP2.6. RESULTS: Compared to the control, elevated CO(2) accelerated phenological progression of tadpoles through Gosner stages, thus resulting in lower body size at metamorphosis. Both elevated temperatures significantly delayed the development and reduced the growth of tadpoles. 100% mortality was observed in 34 °C treatment before metamorphosis (before Gosner stage 36) while all the tadpoles died after metamorphosis (at Gosner stage 46) in 32 °C treatment. Elevated CO(2) increased tadpole activity, in terms of their swimming speed, while both of the elevated temperatures reduced it compared to the control. Catalase activity increased at elevated CO(2). Ammonia excretion by tadpoles was decreased by elevated CO(2), but increased under temperature elevation. Both Elevated CO(2) and temperature treatments reduced the white blood cell count and its percentage of thrombocytes. Percentages of lymphocytes, monocytes and neutrophils were increased at 32 °C, while lymphocyte percentage and lysozyme activity were increased at elevated CO(2). Several deformities were observed in tadpoles at elevated temperature and CO(2). CONCLUSIONS: Elevated temperatures and reduced pH due to elevated CO(2), being major features of climate change, increase the vulnerability of amphibians, who are already one of the most threatened vertebrate groups. Based on our observations on the model amphibian species P. cruciger, increased vulnerability to climate change occurs by reducing their growth, body size and motility while also reducing their immunity and inducing physical deformities. These impacts are highly-likely to reduce the foraging, competitive and reproductive capabilities in their natural habitats. We conclude further that even the ‘best-case’ scenario of future climate change can impose significant physiological impacts that could threaten amphibian populations on broader spatial and temporal scales. GRAPHICAL ABSTRACT: [Image: see text]