Climate change and emerging infectious diseases are often described as the main factors associated with the worldwide amphibian population decline. In this context, rising temperatures due to global warming might act as a chronic stressor for many amphibians, leading to immunosuppression. This study aimed to characterize the thermal sensitivity of the Bullfrog's (Lithobates catesbeianus) immune response and the effect of acclimation at different temperatures on it. Plasma bacterial killing ability (BKA) and phagocytosis activity of blood leukocytes were measured at different incubation temperatures (5–40°C) in individuals kept at 28°C and 34°C. First, all individuals were held under 28°C and sampled on the 16th day. Subsequently, one group was kept at 28°, and the other one was transferred to 34°C. Both groups were sampled at 83 and 106 days of maintenance. Plasma corticosterone (CORT) and testosterone (T) were assessed to evidence thermal stress and possible endocrine correlates of immune changes over time. The incubation temperature affected BKA both on animals kept at 28°C and 34°C, with maximum values at lower temperatures (5–20°C). Phagocytosis activity was constant over the range of assay temperatures. Immune and endocrine variables decreased over time in both thermal regimes, but frogs maintained at 34°C showed lower T and immunosuppression, evidencing stress response. Therefore, exposure to high temperatures might decrease immune function in bullfrogs due to chronic stress response and by exposition to temperatures of lower performance according to the thermal sensitivity curve, which might increase vulnerability to diseases in this anuran species.

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Currently, effects of increased atmospheric temperature, in the context of ongoing climate change, have been investigated in multiple organisms and levels of biological organization. While there has been a focus on the impacts of increased mean temperature, an emergent and equally important point is the consequences of recurrent exposure to extreme temperature events, simulating heat waves. This study investigated the effects of serial exposure to high temperatures on immune and endocrine variables before and after exposure to an acute secondary stressor in bullfrogs (Lithobates catesbeianus). Adult males were divided into three groups and subjected to three thermal regimes: Control (C; constant 22°C); Experimental 1 (E1; kept at 22°C and exposed to 4 days of 30°C every 16 days); and Experimental 2 (E2; kept at 22°C and exposed to 4 days of 30°C every 6 days). Blood samples were collected on the last day of key extreme heat events. Two weeks after the last extreme heat event, animals were subjected to restraint stress (1h) and sampled again. Blood samples were used to determine neutrophil: lymphocyte ratio, plasma bacterial killing ability, as well as corticosterone and plasma testosterone levels. Overall, we found exposure to extreme heat events did not affect immune and endocrine variables over time. Meanwhile, the previous exposure to extreme heat events modulated the responsiveness to restraint. The amplitude of increased corticosterone plasma levels and neutrophil: lymphocyte ratio in response to restraint decreased with the number of previous exposures to extreme heat events. These results suggest that exposure to extreme climatic events has hidden effects on bullfrog's stress response, expressed as diminished reactive scope to a novel stressor. This represents a highly deleterious facet of climate change since diminished responsiveness prevents proper coping with wildlife challenges.

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