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. 2025 Jul 2;16(1):93.
doi: 10.1186/s40104-025-01229-2.

Co-cultivation of Lactobacillus acidophilus and Bacillus subtilis mediates the gut-muscle axis affecting pork quality and flavor

Zhixin Lin  1 , Xinchen Zhou  1   2 , Tingting Lu  1 , Wendong An  1   2 , Shenghao Chen  1 , Suchen Li  1 , Hui Miao  1   2 , Xinyan Han  3   4
Affiliations

Co-cultivation of Lactobacillus acidophilus and Bacillus subtilis mediates the gut-muscle axis affecting pork quality and flavor

Zhixin Lin et al. J Anim Sci Biotechnol. .

Abstract

Background: Pork quality and flavor are critical determinants of consumer preference, yet the role of gut microbiota in shaping meat characteristics remains underexplored. In this study, we investigated how a probiotic consortium (FAM: Lactobacillus acidophilus and Bacillus subtilis) modulates the gut-muscle axis to enhance pork flavor.

Results: In finishing pigs, FAM supplementation significantly increased flavor-associated nucleotides and umami-enhancing amino acids in longissimus dorsi muscle. Metagenomic analysis revealed FAM-driven enrichment of glycan-degrading Prevotella and short-chain fatty acid-producing Phascolarctobacterium, accompanied by reduced antibiotic resistance genes and virulence factors. Spearman correlation linked Prevotella copri abundance with elevated muscle amino acids, suggesting microbial-encoded CAZymes as key mediators.

Conclusions: This study provides the first evidence that probiotic-induced gut microbiota remodeling enhances pork flavor through metabolic cross-talk along the gut-muscle axis. The findings suggest a novel strategy for improving pork quality via dietary interventions targeting gut microbiota.

Keywords: Gut-muscle axis; Metagenome; Pork flavor; Pork quality; Probiotic.

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: The animal experiment was approved by the Animal Care and Use Committee of Zhejiang University (permit number: ZJU20230042) and all experimental procedures conformed to the institutional guidelines for animal study. All efforts were made to minimize suffering. Consent for publication: Not applicable. Competing interests: The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Muscle fiber morphology HE staining section (A); Immunofluorescence sections of muscle fibers (B). CON group, basal diet; FAM group, basal diet added with 0.1% FAM
Fig. 2
Fig. 2
Content of flavor-presenting nucleotides in pig’s longissimus dorsi (n = 6). CON group, basal diet; FAM group, basal diet added with 0.1% FAM. AMP: adenosine monophosphate; GMP: guanosine 5'-monophosphate; IMP: inosine 5'-monophosphate
Fig. 3
Fig. 3
Content of long-chain fatty acids in pig’s longissimus dorsi (n = 6). CON group, basal diet; FAM group, basal diet added with 0.1% FAM
Fig. 4
Fig. 4
Diversity and structure of pig colonic microbiota. A Venn diagram of species numbers. B Alpha diversity index of the colonic microbiota. C Beta diversity analysis of the colonic microbiota. D Community structure of gut bacteria on Phylum level. E community structure of gut bacteria on Species-level. F LEfSe analysis. CON group, basal diet; FAM group, basal diet added with 0.1% FAM
Fig. 5
Fig. 5
Analysis of COG function in colonic microbiota. A–C Wilcoxon rank-sum test from category to COG. D and E Linear discriminant analysis from function to COG, LDA > 2. F Correlation heatmap between gut bacterial signature species and COG function
Fig. 6
Fig. 6
Analysis of KEGG function in colonic microbiota. A–C Wilcoxon rank-sum test of level 1–3. D–F Linear discriminant analysis of level 1–3, LDA > 2. G Correlation heatmap between gut bacterial signature species and KEGG function
Fig. 7
Fig. 7
Analysis of CAZy function in colonic microbiota. A and B Wilcoxon rank-sum test from Class to Family. C Linear discriminant analysis of Family, LDA > 2.5. D Correlation heatmap between gut bacterial signature species and CAZy families
Fig. 8
Fig. 8
Analysis of ARDB and VFDB in colonic microbiota. A and B Wilcoxon rank-sum test from Antibiotic class to ARDB. C and D Linear discriminant analysis from Antibiotic class to Antibiotic type, LDA > 2. E and F Linear discriminant analysis from level 2 to VFs, LDA > 2; G and H Wilcoxon rank-sum test from level 2 to VFs
Fig. 9
Fig. 9
Correlation heatmap between colonic microbiota and muscle amino acid content
See this image and copyright information in PMC

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