Hypoxic Preconditioning Enhances the Hypoxia Tolerance of the Pearl Oyster Pinctada fucata martensii and Is Associated with Changes in the Intestinal Microbiota

Hypoxia frequently triggers mass mortality events in pearl oysters during the summer months. Hypoxic preconditioning (HP), repeated exposure to sublethal low-oxygen conditions, has been proposed as a potential strategy to enhance stress resistance. Here, we investigated how HP affects hypoxia tolerance in the pearl oyster Pinctada fucata martensii, with emphasis on host apoptotic and immune regulation and the gut microbiota. Pearl oysters assigned to HP (experimental group, EG) and to a non-preconditioned control group (CG) were subjected to sustained hypoxic challenge (1.5 ± 0.1 mg/L DO for 15 days). HP significantly increased the expression of apoptosis- and immunity-related genes (MyD88, IκK, NF-κB) while suppressing JNK expression in gills after extended hypoxia (MyD88: EG 2.26 ± 0.65 vs. CG 0.96 ± 0.29, p < 0.05, ~2.3-fold increase; NF-κB: EG 1.50 ± 0.20 vs. CG 0.81 ± 0.31, p < 0.05, ~1.8-fold increase; IκK: EG 1.55 ± 0.38 vs. CG 0.65 ± 0.12, p < 0.05, ~4.0-fold increase; JNK: EG 0.49 ± 0.25 vs. CG 1.44 ± 0.51, p < 0.05, ~0.34-fold), consistent with a pre-activated yet controlled stress response. In parallel, HP markedly reshaped the intestinal microbial community under hypoxia, increasing alpha diversity (Ace, Chao, and Sobs indices) and enriching potentially beneficial bacterial phyla such as Planctomycetota, Nitrospirota, and Fusobacteriota, groups often linked to nutrient cycling and short-chain fatty acid production. Collectively, these results suggest that HP-enhanced hypoxia tolerance in P. f. martensii is associated with coordinated modulation of host apoptotic and immune signaling and concomitant shifts in gut microbiome diversity. These findings highlight the role of the host–microbiota axis in environmental acclimation and suggest that HP may be a practical tool for improving bivalve performance under hypoxic stress in aquaculture.