Heat hardening of a larval amphibian is dependent on acclimation period and temperature.

Plasticity in heat tolerance provides ectotherms the ability to reduce overheating risk during thermal extremes. However, the tolerance-plasticity trade-off hypothesis states that individuals acclimated to warmer environments have a reduced plastic response, including hardening, limiting their ability to further adjust their thermal tolerance. Heat hardening describes the short-term increase in heat tolerance following a heat shock that remains understudied in larval amphibians. We sought to examine the potential trade-off between basal heat tolerance and hardening plasticity of a larval amphibian, Lithobates sylvaticus, in response to differing acclimation temperatures and periods. Lab-reared larvae were exposed to one of two acclimation temperatures (15°C and 25°C) for either 3 or 7 days, at which time heat tolerance was measured as critical thermal maximum (CT ). A hardening treatment (sub-critical temperature exposure) was applied 2 h before the CT assay for comparison to control groups. We found that heat-hardening effects were most pronounced in 15°C acclimated larvae, particularly after 7 days of acclimation. By contrast, larvae acclimated to 25°C exhibited only minor hardening responses, while basal heat tolerance was significantly increased as shown by elevated CT temperatures. These results are in line with the tolerance-plasticity trade-off hypothesis. Specifically, while exposure to elevated temperatures induces acclimation in basal heat tolerance, shifts towards upper thermal tolerance limits constrain the capacity for ectotherms to further respond to acute thermal stress.