The difference and variation of gut bacterial community and host physiology can support adaptation during and after overwintering in frog population.

The hibernation of amphibians can offer a unique window into overwintering adaptation processes and host-gut microbiota interactions through changes in metabolic availability and homeostasis. We attempted to identify differences in the physiology and gut microbiome during and after hibernation in Japanese wrinkled frogs (Glandirana rugosa), an aquatic overwintering amphibian. After hibernation, the high alpha and beta diversity of the gut bacterial community appears to reflect the more diverse and complex environmental conditions. During winter, Proteobacteria dominated the majority of the gut bacterial community, likely due to high oxygen saturation. After hibernation, Firmicutes and Bacteroidetes increased, which are supportive of host metabolism by gut microbiota. Corticosterone also showed high values and variances after hibernation, presumably allowing the population to remain adaptable across a broad range of environmental gradients. Innate immunity was high after hibernation but exhibited low variation among populations, which supports the idea of a prioritized investment in immunity after hibernation. Blood biochemistry suggests that aquatic overwintering frogs have a mechanism to adapt through overhydration and regulate homeostasis through water excretion associated with the kidney and urine after hibernation. Frog populations exhibit variations and adaptability in gut microbiota and physiology during and after hibernation: Through this, they may demonstrate an adaptive response that regulates metabolic availability in preparation for unpredictable environmental changes. We also propose that the maintenance of Proteobacteria during hibernation can support the colonization of Firmicutes and Bacteroidetes after hibernation, underscoring the need to study the complex effects of gut microbiota across multiple life stages.