Captivity-driven gut dysbiosis in leopards (Panthera pardus): effects of diet and rearing practices

India is a biodiversity hotspot and supports a vast diversity of life forms including large cats. Among large cats, leopards are distributed widely across geographical terrains of the country. Owing to close proximity to human settlements and persistent conflict situation, many leopards are rescued due to conflict, injury or orphaning and are subsequently maintained under long term captivity or hand reared conditions. In wild, the leopards feed on a diverse prey base; however, in captivity the diet and feeding pattern remain uniform. The impact of such dietary simplification and captive management on gut microbiome composition remains poorly understood. In the current study, the gut microbiome of wild, captive, and hand-raised leopards was evaluated and compared to estimate the microbial diversity and bioinformatics analysis was carried out to assess microbial diversity, taxonomic composition and differential abundance. A total of 22 fresh scat samples were collected, including samples from wild ( n = 9), captured captive ( n = 8), and hand-raised captive leopards ( n = 5). Two wild samples (W5 and W8) were excluded from diversity analyses due to failed genotyping, yielding 20 samples (wild n = 7, captive n = 8, hand-raised n = 5) for downstream ecological analysis. Beta diversity and multivariate analyses exhibited clear separation between wild and captive groups, highlighting strong effects captive management of gut microbial structure. Taxonomic analysis indicated dominance of Bacteroidetes and Firmicutes in wild leopards, whereas captive and hand-raised leopards showed increased abundance of Firmicutes and Proteobacteria, including several taxa associated with dysbiosis. Differential abundance analysis identified specific bacterial species as potential indicators of captivity-related microbial alteration. These findings demonstrate that captivity, particularly long-term confinement and hand-rearing with uniform diets, significantly alters the gut microbiome of leopards. The study emphasizes the importance of dietary diversification and microbiome-informed management strategies to improve health and welfare of captive leopards and to enhance conservation outcomes.