Deciphering the Mechanism by Which Carbon Dioxide Extends the Shelf Life of Raw Milk: A Microbiomics- and Metabolomics-Based Approach.

Microbial community succession in raw milk determines its quality and storage period. In this study, carbon dioxide (CO2) at 2000 ppm was used to treat raw milk to investigate the mechanism of extending the shelf life of raw milk by CO2 treatment from the viewpoint of microbial colonies and metabolites. The results showed that the shelf life of CO2-treated raw milk was extended to 16 days at 4 °C, while that of the control raw milk was only 6 days. Microbiomics analysis identified 221 amplicon sequence variants (ASVs) in raw milk, and the alpha diversity of microbial communities increased (p < 0.05) with the extension of storage time. Among them, Pseudomonas, Actinobacteria and Serratia were the major microbial genera responsible for the deterioration of raw milk, with a percentage of 85.7%. A combined metagenomics and metabolomics analysis revealed that microorganisms altered the levels of metabolites, such as pyruvic acid, glutamic acid, 5'-cmp, arginine, 2-propenoic acid and phenylalanine, in the raw milk through metabolic activities, such as ABC transporters, pyrimidine metabolism, arginine and proline metabolism and phenylalanine metabolism, and reduced the shelf life of raw milk. CO2 treatment prolonged the shelf life of raw milk by inhibiting the growth of Gram-negative aerobic bacteria, such as Acinetobacter guillouiae, Pseudomonas fluorescens, Serratia liquefaciens and Pseudomonas simiae.

Characterization and Discrimination of Volatile Compounds in Chilled Tan Mutton Meat during Storage Using HiSorb-TD-GC-MS and E-Nose.

Chilled Tan mutton is currently the mainstream of Tan mutton production and consumption in China, but the reports on chilled meat quality evaluation and shelf-life discrimination by volatiles are limited. This study aimed to investigate the changes of volatile compounds in chilled Tan mutton at four storage stages (1d, 3d, 5d, 7d) in order to differentiate the various storage stages. An analysis protocol was established for the characterization and discrimination of the volatiles in chilled Tan mutton based on high capacity sorptive extraction-thermal desorption-gas coupled with chromatography-mass spectrometry (HiSorb-TD-GC-MS), electronic nose (E-nose), and multivariate statistical analysis. A total of 96 volatile compounds were identified by HiSorb-TD-GC-MS, in which six compounds with relative odor activity value >1 were screened as the key characteristic volatiles in chilled Tan mutton. Four storage stages were discriminated by partial least squares discriminant analysis, and nine differential volatile compounds showed a variable importance for the projection score >1, including octanoic acid, methyl ester, decanoic acid, methyl ester, acetic acid, heptanoic acid, methyl ester, propanoic acid, 2-hydroxy-, methyl ester, (ñ)-, hexanoic acid, propanoic acid, butanoic acid, and nonanoic acid. With the volcano plot analysis, hexadecanoic acid, methyl ester, was the common volatile marker candidate to discriminate chilled stages of Tan mutton. Meanwhile, E-nose could discriminate chilled Tan mutton at different storage stages rapidly and efficiently using linear discriminant analysis. Furthermore, E-nose sensors could obtain comprehensive volatile profile information, especially in esters, acids, and alcohols, which could confirm the potential of E-nose for meat odor recognition. Thus, this analysis protocol could characterize and discriminate the volatiles in chilled Tan mutton during storage.

Footprint analysis of CO2 in microbial community succession of raw milk and assessment of its quality.

With the growing production of raw milk, interest has been increasing in its quality control. CO2, as a cold processing additive, has been studied to extend the cold storage period and improve the quality of raw milk. However, it is yet uncertain how representative microbial species and biomarkers can succeed one another at distinct critical periods during refrigeration. Therefore, the effects of CO2 treatment on the succession footprint of the microbial community and changes in quality during the period of raw milk chilling were examined by 16S rRNA analysis combined with electronic nose, and electronic tongue techniques. The results indicated that, the refrigeration time was shown to be prolonged by CO2 in a concentration-dependent way. And CO2 treatment was linked to substantial variations in beta and alpha diversity as well as the relative abundances of various microbial taxa (p < 0.01). The dominant bacterial phylum Proteobacteria was replaced with Firmicutes, while the major bacterial genera Acinetobacter and Pseudomonas were replaced with lactic acid bacteria (LAB), including Leuconostoc, Lactococcus, and Lactobacillus. From the perspective of biomarkers enriched in CO2-treated sample, almost all of them belong to LAB, no introduction of harmful toxins has been found. The assessment of the quality of raw milk revealed that CO2 improved the quality of raw milk by lowering the acidity and the rate of protein and fat breakdown, and improved the flavor by reducing the generation of volatiles, and increasing umami, richness, milk flavor and sweetness, but reducing sourness. These findings offer a new theoretical foundation for the industrial use of CO2 in raw milk.