Shotgun Metagenomic Analysis of Gut Microbiota and Antibiotic Resistance Genes in a High-Fat Diet Mouse Model Treated with Heat-Killed Lactiplantibacillus plantarum beLP1

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The gut microbiota is a central regulator of metabolic function, and its disruption by a high-fat diet (HFD) is strongly linked to obesity and metabolic impairment. This study evaluated the potential of heat-killed Lactiplantibacillus plantarum beLP1 (beLP1®) in alleviating HFD-induced metabolic and microbial imbalances in mice. Male C57BL/6N mice were fed an HFD for 10 weeks, with or without daily oral supplementation of beLP1 (≥3 × 1010 cells). Compared with untreated HFD mice, beLP1 supplementation reduced serum triglycerides by 35% and lowered liver enzymes AST and ALT by 17% and 36%, respectively. Blood glucose levels remained similar to the HFD group throughout the study period. Shotgun metagenomic analysis revealed that beLP1 restored gut microbial diversity, increased beneficial taxa such as Akkermansia and Faecalibaculum high. and reduced pro-inflammatory species including Streptococcus sp., Mucispirillum schaedleri and Clostridium cocleatum. These microbial changes were associated with partial normalization of the Firmicutes/Bacteroidota ratio and improvements in antibiotic resistance gene (ARG) profiles. Specifically, in silico analysis of the short-chain fatty acid (SCFA) synthesis pathways indicated that the potential for acetate and propionate production was maximized in the beLP1 group, resulting in the highest relative abundance among all groups. This functional enhancement directly correlated with the enrichment of key SCFA-producing taxa, particularly Akkermansia muciniphila, confirming that increased bacterial abundance suggests an enhanced functional potential for SCFA production. Furthermore, beLP1® induced a selective modulation of gut ARGs, significantly reducing specific subtypes such as tetracycline and multidrug efflux genes, despite a slight increase in vancomycin resistance markers. Overall, our findings suggest that beLP1® attenuated the rate of body weight gain during the initial weeks of HFD exposure and significantly improved markers of hepatic stress and lipid metabolism.

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