Unraveling the spoilage characteristics of refrigerated pork using high-throughput sequencing coupled with UHPLC-MS/MS-based non-targeted metabolomics

The spoilage of refrigerated pork involves nutrient depletion and the production of spoilage metabolites by spoilage bacteria, yet the microbe-metabolite interactions during this process remain unclear. This study employed 16S rRNA high-throughput sequencing and non-targeted metabolomics based on ultra-highperformance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to reveal the core microbiota and metabolite profiles of pork during refrigeration. A total of 45 potential biomarkers were screened through random forest model analysis. Metabolic pathway analysis indicated that eleven pathways, including biogenic amine metabolism, pentose metabolism, purine metabolism, pyrimidine metabolism, phospholipid metabolism, and fatty acid degradation, were potential mechanisms of pork spoilage. Correlation analysis revealed nine metabolites-histamine, tyramine, tryptamine, D-gluconic acid, UDP-D-glucose, xanthine, glutamine, phosphatidylcholine, and hexadecanoic acid-as spoilage biomarkers, with Pseudomonas, Serratia, and Photobacterium playing significant roles. This study provides new insights into the changes in microbial and metabolic characteristics during the spoilage of refrigerated pork.