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Earthworms and mealworms are widely used as the primary biological feeds of Quasipaa spinosa . In the present study, a controlled feeding trial was conducted to evaluate the effects of three different biological feed types—earthworm feed (EF), mealworm feed (TM), and mixed feed (MIX)—on the growth performance, antioxidant enzyme activity, digestive enzyme activity, and gut microbial diversity of Q. spinosa to determine the optimal biological feeding strategy. The results demonstrated that frogs in the MIX group exhibited the highest final body weight (FBW), survival rate (SR), weight gain rate (WGR), and specific growth rate (SGR), all of which were significantly higher than those observed in the TM group ( P < 0.05). Our study revealed that superoxide dismutase (SOD) activity was highest in the TM group, whereas catalase (CAT) activity was the lowest in the same group. In contrast, digestive enzyme activities, including trypsin, amylase, and lipase were consistently highest in the MIX group, with trypsin and amylase activities being significantly higher than those in the EF and TM groups ( P < 0.05). In addition, high-throughput sequencing identified a total of 4020 operational taxonomic units (OTUs) in the intestinal microbiota, of which 1154 were shared among all groups. The MIX group exhibited the largest number of unique OTUs (745). At the phylum level, Bacillota (Firmicutes) and Pseudomonas (Proteobacteria) were the predominant taxa across all treatments, with Bacillota emerging as a core phylum in the intestines of Q. spinosa . At the genus level, Romboutsia and Clostridium were the dominant, with Romboutsia serving as the core genus and exhibiting the highest relative abundance across all groups. Taken together, these findings indicate that mixed biological feeding enhances growth performance, digestive capacity, and intestinal microbial diversity, thereby contributing to improved intestinal health in Q. spinosa . This study provides experimental evidence that may contribute to the optimization of biological feeding strategies in amphibian aquaculture. • Mixed biological feeding improves growth performance of Q. spinosa . • The MIX diet enhances digestive enzyme activities and antioxidant capacity. • Mixed biological feeding increases gut microbial diversity and promotes a balanced intestinal microbiota composition.