Evidence for a metal disease refuge: The amphibian-killing fungus (Batrachochytrium dendrobatidis) is inhibited by environmentally-relevant concentrations of metals tolerated by amphibians

The amphibian-killing fungus Batrachochytrium dendrobatidis (Bd) has caused substantial declines in Bd-susceptible amphibian species worldwide. However, some populations of Bd-susceptible frogs have managed to survive at existing metal-polluted sites, giving rise to the hypothesis that frogs might persist in the presence of Bd if Bd is inhibited by metals at concentrations that frogs can tolerate. We tested this hypothesis by measuring the survival of Bd zoospores, the life stage that infects amphibians, and calculated the LC 50 after exposure to environmentally-relevant elevated concentrations of copper (Cu), zinc (Zn), and their combination (Cu+Zn) in two repeated 4-day acute exposure runs. We also measured the chronic sensitivity of Bd to these metals over three generations by measuring the number of colonies and live zoospores and calculating EC 50 concentrations after 42 days of exposure. We then compared acute and chronic sensitivity of Bd with amphibian sensitivities by constructing species sensitivity distributions (SSDs) using LC 50 and EC 50 data obtained from the literature. Acute sensitivity data showed that Bd zoospore survival decreased with increasing metal concentrations and exposure durations relative to the control, with the highest LC 50 values for Cu and Zn being 2.5 μg/L and 250 μg/L, respectively. Chronic exposures to metals resulted in decreased numbers of Bd colonies and live zoospores after 42 days, with EC 50 values of 0.75 μg/L and 1.19 μg/L for Cu and Zn, respectively. Bd zoospore survival was 10 and 8 times more sensitive to Cu and Zn, respectively in acute, and 2 and 5 times more sensitive to Cu and Zn in chronic exposure experiments than the most sensitive amphibian species recorded. Our findings are consistent with the hypothesis that metals in existing metal-polluted sites may have a greater impact on Bd relative to amphibians' performance, potentially enabling Bd-susceptible amphibians to persist with Bd at these sites.