Enterotoxigenic Escherichia coli (ETEC) causes severe intestinal infections in animals and threatens public health under the One Health framework. Most conventional studies focus on acute short-term ETEC infection, while natural persistent colonization oftern induces chronic intestinal mucosal compensatory remodeling in hosts. This study evaluated the protective effects of giant panda-derived Weissella confusa BSP201703 against chronic ETEC-induced intestinal damage using a giant panda fecal microbiota-associated (GPF) mouse model. Seventy-two Kunming mice were divided into six groups: blank control (C1), GPF control (C2), ETEC control (C3), and three W. confusa BSP201703 groups at low (1.0 × 107 cfu/mL, W1), medium (1.0 × 108 cfu/mL, W2), and high (1.0 × 109 cfu/mL, W3) doses. Mice were first subjected to continuous ETEC challenge for 5 days to establish stable chronic intestinal injury, followed by a subsequent 5-day intervention with probiotic or sterile PBS for repairing existing damage. Growth performance, histopathology, serum D-lactate, SIgA, tight junction genes (ZO-1, Occludin, Claudin-1), and gut microbiota were analyzed. Histomorphologically, the chronic ETEC challenge induced compensatory increases in ileal villus height and crypt depth, which differed from typical acute necrotic atrophy. W. confusa BSP201703 mitigated ETEC-induced damage, reduced serum D-lactate (p < 0.05), increased SIgA, and upregulated tight junctions (p < 0.05). Microbial results demonstrated that medium-dose W2 maximized microbial diversity, while W1/W3 selectively enriched beneficial Bacteroidetes, Clostridium cluster IV, and Clostridium cluster XIVa taxa, confirming that moderate doses yielded optimal protection. In conclusion, W. confusa BSP201703 relieves ETEC injury by enhancing intestinal barrier function and regulating gut microbiota, highlighting its potential as a wildlife probiotic for One Health applications.
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