Effects of Mobile Sheepfold and Supplementary Feeding on Growth Performance, Serum Indicators and Gut Microbiota in Natural Grazing Gangba Sheep

High-altitude grazing animals are continuously exposed to strong wind and low temperature, which challenge physiological homeostasis and energy metabolism. Improving living conditions and nutritional supplementation are two commonly used strategies. In this study, sixty 7-month-old Gangba sheep (initial body weight (BW) 21.00 ± 1.90 kg) were allocated to a 42-day trial with four groups (open-air sheepfold, mobile sheepfold, open-air sheepfold + supplementary feeding, mobile sheepfold + supplementary feeding) to investigate their effects on growth performance, serum parameters and gut microbiota in naturally grazing Gangba sheep. Mobile sheepfolds increased the temperature-humidity index (THI) and reducing the wind chill index (WCI) (<i>p</i> < 0.05). The sheep with mobile sheepfold showed higher serum total antioxidant capacity and lower levels of heat shock proteins HSP70 and HSP90 (<i>p</i> < 0.05), indicating alleviated stress. Supplementary feeding markedly increased final BW and average daily gain (<i>p</i> < 0.05). The interaction between sheepfold type and feeding supplementation showed increasing IgA levels in the open-air sheepfold with supplementary feeding group and increasing IL-4 levels in the mobile sheepfold with supplementary feeding group, while TNF-α concentrations were reduced in all three treatment groups (<i>p</i> < 0.05). Meanwhile, KB and FFAs were increased in the open-air sheepfold with supplementary feeding group but decreased in the mobile sheepfold with supplementary feeding group (<i>p</i> < 0.05). The mobile sheepfold also increased the Bacillota-to-Bacteroidota ratio, suggesting improved microbial community structure. Functional predictions showed enrichment of reductive acetogenesis and reduction in aerobic chemoheterotrophy and sulfur-related respiration pathways (<i>p</i> < 0.05). Moreover, key microbial genera were significantly correlated with THI and WCI (<i>p</i> < 0.05). Collectively, these results demonstrated that mobile sheepfold together with feeding supplementation improve stress responses, serum immune and lipid metabolic indicators, and potentially altered gut microbial composition and function, providing insights into host-microbiota interaction in extreme high-altitude environments.