Quasipaa spinosa, commonly known as the spiny frog, is an economically valued amphibian in China prized for its tender meat and nutritional value. This species exhibits marked sexual dimorphism, most notably the prominent spiny structures on males that are pivotal for mating success and species identification. The spines of Q. spinosa exhibit strong seasonal variation, changing significantly with the reproductive cycle, which typically spans from April to October. Sexually mature males develop densely packed, irregularly arranged round papillae with black spines on their chests during the breeding season, which may then reduce or disappear afterward, while females have smooth chest skin. Despite their ecological importance, the developmental mechanisms and biological functions of these spines have been inadequately explored. This study integrates morphological, transcriptomic, and metabolomic analyses to elucidate the mechanisms underlying the seasonal variation in spine characteristics of Q. spinosa. Our results demonstrate that spine density inversely correlates with body size and that spine development is accompanied by significant changes in epidermal thickness and keratinization during the breeding season. Comparative transcriptomic analysis across different breeding stages revealed significant gene expression alterations in pathways related to extracellular matrix interactions, tyrosine metabolism, Wnt signaling, and melanogenesis. Metabolomic analysis further identified significant seasonal shifts in metabolites essential for energy metabolism and melanin synthesis, including notable increases in citric acid and β-alanine. These molecular changes are consistent with the observed morphological adaptations, suggesting a complex regulatory mechanism supporting spine development and functionality. This study provides novel insights into the molecular basis of spine morphogenesis and its seasonal dynamics in Q. spinosa, contributing valuable information for the species’ conservation and aquaculture.

The Chinese giant salamander (CGS, Andrias davidianus) is an endangered aquatic amphibian in China. Using environmental DNA (eDNA), this study aimed to monitor the spatial distribution of this species in Zhejiang Province, providing a scientific basis for species assessment, evaluation of reintroduction effectiveness, and non-invasive monitoring. Based on historical records of species presence, 18 representative water localities were selected for sampling. Among these localities, eDNA signals of the CGS were detected at nine. After excluding sampling sites affected by historical CGS releases and surrounding breeding farms, Jinzi Peak and Huyuan were inferred as candidate potentially natural stream habitats within the province. Additionally, eDNA monitoring was conducted at two release sites, Gutian Mountain and Banshanyuan, for two consecutive years. Results revealed lower detection rates in both areas in the second year of monitoring, potentially attributed to low survival rates, dispersal and disappearance, or sampling variability. A principal component analysis (PCA) of water quality factors (including parameters such as NH₃-N, TN, TP, COD) revealed distinct geographic clustering of CGS-positive and CGS-negative sampling sites that corresponded to their separation in topographic space, suggesting that the measured physicochemical conditions may be associated with the CGS eDNA detection probability. This study demonstrates that eDNA is a feasible and efficient method for surveying CGS in Zhejiang Province. Further, it provides an efficient non-invasive monitoring approach for endangered amphibians, with broad application prospects.