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The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Probiotics are increasingly used as starter cultures to produce fermented dairy products; however, few studies have investigated the role of probiotics in milk fermentation metabolism. The current study aimed to investigate whether adding Bifidobacterium animalis ssp. lactis Probio-M8 (Probio-M8) as a starter culture strain could improve milk fermentation by comparing the physicochemical characteristics and metabolomes of fermented milks produced by a commercial starter culture with and without Probio-M8. Our results showed that adding Probio-M8 shortened the milk fermentation time and improved the fermented milk texture and stability. Metabolomics analyses revealed that adding Probio-M8 affected mostly organic acid, AA, and fatty acid metabolism in milk fermentation. Targeted quantitative analyses revealed significant increases in various metabolites related to the sensory quality, nutritive value, and health benefits of the probiotic fermented milk, including 5 organic acids (acetic acid, lactic acid, citric acid, succinic acid, and tartaric acid), 5 EAA (valine, arginine, leucine, isoleucine, and lysine), glutamic acid, and 2 essential fatty acids (α-linolenic acid and docosahexaenoic acid). Thus, applying probiotics in milk fermentation is desirable. This study has generated useful information for developing novel functional dairy products. [Display omitted] [ABSTRACT FROM AUTHOR]

Probiotic functional products have drawn wide attention because of their increasing popularity. However, few studies have analyzed probiotic-specific metabolism in the fermentation process. This study applied UPLC-QE-MS-based metabolomics to track changes in the milk metabolomes in the course of fermentation by two probiotic strains, Lacticaseibacillus paracasei PC-01 and Bifidobacterium adolescentis B8589. We observed substantial changes in the probiotic fermented milk metabolome between 0 and 36 h of fermentation, and the differences between the milk metabolomes at the interim period (36 h and 60 h) and the ripening stage (60 h and 72 h) were less obvious. A number of time point-specific differential metabolites were identified, mainly belonging to organic acids, amino acids, and fatty acids. Nine of the identified differential metabolites are linked to the tricarboxylic acid cycle, glutamate metabolism, and fatty acid metabolism. The contents of pyruvic acid, γ-aminobutyric acid, and capric acid increased at the end of fermentation, which can contribute to the nutritional quality and functional properties of the probiotic fermented milk. This time-course metabolomics study analyzed probiotic-specific fermentative changes in milk, providing detailed information of probiotic metabolism in a milk matrix and the potential beneficial mechanism of probiotic fermented milk. [ABSTRACT FROM AUTHOR]

Probiotic fermented milk is more and more popular due to their positive health associated properties. However, fermentation temperature and other process conditions may affect the growth and metabolism of probiotic strains, thereby affecting quality of the final products. In this study, the growth behaviors and metabolomic profiles of yogurts induced by Lactobacillus casei Zhang at fermentation termination (FT) and d 10 of storage (S10d) under different fermentation temperatures at 37°C (low) and 42°C (high) were analyzed and compared using liquid chromatography-mass spectrometry (MS)- and gas chromatography-MS-based metabolomics approaches. At 37°C, the growth of L. casei Zhang at FT and S10d was significantly increased, and the potential relationship between riboflavin, starch, and sucrose metabolism and growth of L. casei Zhang may be mutually promoting. Fermentation temperature (37°C and 42°C) affected volatile and nonvolatile metabolomic profiles and pathways. The levels of acetaldehyde, 2,3-butanedione, acetoin, butyric acid, decanoic acid, hexanoic acid, and octanoic acid were significantly higher at 37°C than at 42°C at FT and S10d. This indicates that the low temperature (37°C) most likely contributes more to the formation of important flavor compounds during the fermentation process and production of short-chain fatty acids during storage. [Display omitted] [ABSTRACT FROM AUTHOR]

Interest has been growing in the co-fermentation of starter cultures with probiotic bacteria in milk. However, the representative metabolites and metabolic changes at different key time points during milk fermentation and storage in starter cultures and probiotic bacteria are still unclear. In this study, we used gas chromatography/mass spectrometry–based metabolomics to identify volatile metabolites and discriminate between 6 different time points [fermentation initiation (FI), fermentation curd (FC), fermentation termination (FT), storage 1 d (S1d), storage 7 d (S7d), and storage 14 d (S14d)] during the fermentation and storage of starter cultures and Lactobacillus casei Zhang milk. Of the 52 volatile metabolites identified, 15 contributed to discrimination of the 6 time points. Then, using the profile from the different time points, we analyzed pairwise comparisons (FI vs. FC; FC vs. FT; FT vs. S1d; S1d vs. S7d; S7d vs. S14d); these time-lapse comparisons showed metabolic progressions from one fermentation stage to the next. We found representative and exclusive metabolites at specific fermentation and storage time points. The greatest difference in metabolites occurred between FC and FT, and the metabolic profiles between S7d and S14d were most similar. Interestingly, decanoic acid, octanoic acid, and hexanoic acid reached their highest level at storage 14 d, indicating that the post-fermentation storage of fermented milk with L. casei Zhang may add more probiotic functions. This work provides detailed insight into the time-specific profiles of volatile metabolites and their dynamic changes; these data may be used for understanding and eventually predicting metabolic changes in milk fermentation and storage, where probiotic strains may be used. [ABSTRACT FROM AUTHOR]

[Display omitted] • High viable counts of PC-01 and B8589 over storage of fermented milk beverages. • Co-fermentation improves post-storage stability of fermented milk beverages. • Co-fermentation increases γ-aminobutyric acid and malate in stored products. Few studies investigated the effects of co-fermentation with bifidobacteria on post-storage changes of probiotic fermented beverages (PFBs). Thus, this study compared the post-storage changes in physicochemical index and metabolomes of PFBs produced singly by Lacticaseibacillus paracasei PC-01 (PC-01) or in combination with Bifidobacterium adolescentis B8589 (B8589). No significant differences were observed in the pH, titratable acidity, and viable cell counts between the two PFBs over 30-day storage. However, adding B8589 not only increased the stability of PFB (based on evaluating differences in PFBs metabolomics), but also the contents of beneficial amino acid metabolites, including 4-hydroxystyrene, gamma-aminobutyric acid, N -acetyl- l -aspartic acid, d -alanyl- d -alanine, and l -malic acid, after storage. Our study showed that B8589 is preferred to single-strain fermentation by PC-01. This study supports the concept of using bifidobacteria as starter culture in PFB production. [ABSTRACT FROM AUTHOR]

Probiotics are increasingly used in the fermentation industry owing to their probiotic properties. However, most previous studies of milk fermentation are based on single-strain fermentation or focused on interactions between the starter strains without considering the role of probiotics in the microbial ecosystem. Thus, in this study, we compared the differences in the fermented milk metabolomes of single-strain fermentation with Lacticaseibacillus paracasei PC-01 (PC-01) versus dual-strain fermentation with both PC-01 and a probiotic, Bifidobacterium adolescentis B8589 (B8589), with the aim of revealing the metabolic effect of adding B8589 in the milk fermentation process. Although co-fermenting with B8589 did not influence the growth of PC-01, non-targeted metabolomics revealed significant changes in the dual-strain fermented milk metabolomes, characterized by increased levels of amino acid metabolites. Targeted metabolomics analysis confirmed that, by adding B8589 as the co-fermentation culture, the level of γ-aminobutyric acid increased by seven times; and the contents of acetate, propionate, butyrate, valerate, and citrate were also significantly enriched. Our results suggested that B8589 metabolically interacts with PC-01, which increases the amounts of functional metabolites in the co-fermented milk. This study provides interesting insights into the metabolic interaction between probiotic and starter culture in an industrial fermentation process. • Dual-strain is better than single-strain in cow milk fermentation. • Co-fermenting with B8589 changes the fermented milk metabolome. • Co-fermenting with B8589 increases γ-aminobutyric acid amounts. • Co-fermenting with B8589 increases short-chain fatty acid amounts. • Milk co-fermentation increases the functional potential of products. [ABSTRACT FROM AUTHOR]

In the ecosystem of spontaneously fermented cow milk, the characteristics and relationship of bacterial communities and nonvolatile components at different scales of geographical distances (provincial, county, and village levels) are unclear. Here, 25 sampling sites from Xin Jiang and Tibet, 2 provinces of China, were selected based on the distribution of spontaneously fermented cow milk and used for metagenomic and metabolomic analysis. At the provincial geographical distance, the same predominant species, Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus , were detected in Xin Jiang and Tibet. Further, the richness of the bacterial composition of samples from Tibet was higher than those from Xin Jiang; specifically, at the species level, 28 species were identified in Tibet samples but only 7 species in Xin Jiang samples. At the provincial geographical level, we detected significant differences in bacterial structure, shown in principal coordinate analysis plots, and significant differences (Simpson index) in bacterial diversity were also detected. However, at the county and village levels, no significant differences were detected in bacterial communities and diversity, but a difference in bacterial compositions was detectable. This indicates that bacterial communities and diversity of spontaneously fermented milk dissimilarity significantly increased with geographic distance. For the nonvolatile component profiles, the partial least squares discriminant analysis plot (R2Y > 0.5 and Q2 > 0.5 for the goodness-of-fit and predictive ability parameter, respectively) showed that samples from different geographical distances (provincial, county, and village) were all separated, which indicated that all the discriminations in nonvolatile components profiles were from different geographical distances. Investigating relationships between lactic acid bacteria and discriminatory nonvolatile components at the county level showed that 9 species were positively correlated with 16 discriminatory nonvolatile components, all species with low abundance rather than the predominant species L. delbrueckii ssp. bulgaricus and Strep. thermophilus , which indicates the importance of the selection of autochthonous nonpredominant bacteria. [ABSTRACT FROM AUTHOR]

This study aimed to compare the efficacy of fermented cow milk and fermented soy milk as protective carriers for the probiotic strain, Lactiplantibacillus plantarum LP-6 (LP-6), during upper gastrointestinal digestion. Using a near-real dynamic in vitro human stomach system, we evaluated the survival of LP-6 in fermented soy milk and fermented cow milk under simulated upper gastrointestinal conditions. Additionally, we conducted metabolomics analysis at different digestion stages to investigate the potential mechanisms of probiotic survival. Our results revealed that soy milk outperformed cow milk as a carrier, significantly improving the survival of LP-6 throughout digestion. The metabolomics analysis revealed notable enrichment in aspartic acid, glutamate, glutamine, phenylalanine metabolism, and ABC transporter proteins in digested fermented soy milk compared to digested fermented cow milk. These changes likely contribute to enhanced acid tolerance and bacterial cell protection in fermented soy milk. Our findings suggest that soy milk is an optimal carrier for LP-6, effectively protecting specific strains from acid stress while ensuring efficient delivery. • L plantarum LP-6 exhibited higher survival rates in fermented soy milk. • Amino acid metabolism may enhance acid tolerance and bacterial cell protection. • Soy milk was a more suitable carrier medium for L plantarum LP-6. [ABSTRACT FROM AUTHOR]

[Display omitted] • Effects of single and binary probiotics on growth and metabolism were compared. • Growth of B. lactis V9 was improved by co-cultivation with L. casei Zhang. • Valine, leucine and isoleucine biosynthesis may contribute to growth of B. lactis V9. • Binary probiotics had additive effect on production of short-chain fatty acids. In the present study, comparative effects of the single and binary probiotics of Lacticaseibacillus casei Zhang (L. casei Zhang) and Bifidobacterium lactis V9 (B. lactis V9) on the growth and metabolomic profiles during milk fermentation and storage has been analyzed using gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–mass spectrometry (LC–MS). The growth of B. lactis V9 can be significantly increased (p < 0.001) by co-cultivation with L. casei Zhang at the end of fermentation and storage 10 days, and valine, leucine and isoleucine biosynthesis may be a major contributor to the growth promotion of B. lactis V9. However, the growth of L. casei Zhang was not affected by co-cultivation with B. lactis V9. There were notable distinctions in volatile and non-volatile metabolomic profiles and pathways between the single and binary probiotics cultures; binary probiotics L. casei Zhang and B. lactis V9 significantly affected the volatile, non-volatile metabolic profiles compared to the single probiotics. The levels of acetic acid, hexanoic acid, butanoic acid and pentanoic acid were significantly higher (p < 0.05) in binary probiotics cultures compared to the single probiotic cultures at the storage 10 days, which indicates that binary probiotics had additive effects on the production of short-chain fatty acids during storage. This work provides a detailed insight into metabolomic profiles and growth that differ between the single and binary probiotics cultures, and it can be helpful to develop probiotic yogurt with high probiotic viability and distinct metabolomic profiles. [ABSTRACT FROM AUTHOR]

There has been a growing interest in cofermentation of starter cultures with probiotics in milk. In this study, we analyzed the effects of adding the single probiotics Lactobacillus casei Zhang (Zhang) or Bifidobacterium animalis ssp. lactis Probio-M8 (M8) or a combination of Zhang and M8 to starter cultures on volatile and nonvolatile metabolomic profiles after 14 d of storage at 4°C and compared using a liquid chromatography-tandem mass spectrometry (LC-MS) and GC-MS-based metabolomics approach. Principal component analysis, heatmap plots, and Spearman correlation results showed that Zhang alone had a greater effect on volatile and nonvolatile metabolomic profiles than M8 alone. The combination of Zhang and M8 had additive effects on the production of metabolites. For volatile metabolites, the levels of acetaldehyde, diacetyl, acetoin, and acetic acid were higher for the combination of Zhang and M8 compared with either single probiotic culture. Significantly increased nonvolatile components induced by adding Zhang were identified were enriched in the galactose, amino- and nucleotide sugar, fructose and mannose, purine, phenylalanine metabolism, and arginine biosynthesis pathways. The metabolism and biosynthesis of starch, sucrose, tyrosine, galactose metabolism, and aminoacyl-tRNA biosynthesis were significantly upregulated by adding the combination of Zhang and M8. This work provides a detailed insight into different effects of Zhang and M8 used alone or in combination on the volatile and nonvolatile metabolomic profiles of yogurts. [ABSTRACT FROM AUTHOR]