Chemical profiling and probiotic viability assessment in Gueuze-style beer: Fermentation dynamics, metabolite and sensory characterization, and in vitro digestion resistance.

Probiotic viability, metabolite concentrations, physicochemical parameters, and volatile compounds were characterized in Gueuze beers formulated with probiotic lactic acid bacteria (LAB) and yeast. Additionally, the sensory profile of the beers and the resistance of the probiotics to digestion were determined. The use of 2 International Bitterness Units resulted in high concentrations of probiotic LAB but a decline in probiotic yeast as pH decreased. Secondary fermentation led to the consumption of maltose, citric acid, and malic acid, and the production of lactic and propionic acids. Carbonation and storage at 4 °C had minimal impact on probiotic viability. The addition of probiotic LAB resulted in a distinct aroma profile with improved sensory characteristics. Our results demonstrate that sour beers produced with probiotic LAB and a probiotic yeast, and fermented using a two-step fermentation process, exhibited optimal physicochemical parameters, discriminant volatile compound profiles, promising sensory characteristics, and high probiotic concentrations after digestion.

Cytotoxicity assessment and antimicrobial effects of cell-free supernatants from probiotic lactic acid bacteria and yeast against multi-drug resistant Escherichia coli.

The antibacterial, antibiofilm, and cytotoxicity activity of cell-free supernatants (CFSs) from probiotics, including Lactobacillus plantarum, Bifidobacterium bifidum, and Saccharomyces cerevisiae against multi-drug resistant Escherichia coli evaluated in current research. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the CFSs were determined by analyzing inhibition zone formation using agar disk diffusion for antibacterial activity, microtiter plate for biofilm analysis, and auto-aggregation were done. CFSs substances were analyzed by GC-MS. The MTT assay on HEK293 cells investigated CFS's influence on cell viability. CFSs were examined for biofilm-related virulence genes, including aggR and fimH using real-time polymerase chain reaction (real-time PCR). All CFSs had bacteriostatic and bactericidal effects. The B. bifidum exhibited the highest antibiofilm activity compared to the others. Bifidobacterium bifidum, L. plantarum, and S. cerevisiae produce 19, 16, and 11 mm inhibition zones against E. coli, respectively. GC-MS indicated that Hydroxyacetone, 3-Hydroxybutyric acid, and Oxime-methoxy-phenyl-dominated CFSs from L. plantarum, B. bifidum, and S. cerevisiae CFSs, respectively. The MTT test demonstrated a cell viability rate of over 90%. Statistically, adding all CFSs lowered the relative expression of both aggR and fimH virulence genes.

Untargeted Metabolomics Analysis of Lactic Acid Bacteria Fermented Acanthopanax senticosus with Regard to Regulated Gut Microbiota in Mice.

Previous studies have shown that Acanthopanax senticosus (AS) has a beneficial preventive and therapeutic effect on colitis. The fermentation of lactic acid bacteria (LAB) can alter the efficacy of AS by modifying or producing new compounds with potential bioactive properties. However, the specific components and mechanisms that enhance the efficacy are still unclear. In the present experiment, untargeted metabolomics was used to analyze the changes in active components before and after LAB fermentation of AS. The aim was to explain the mechanism of AS fermentation in treating colitis using a colitis model in mice. The results indicated that the fermentation of LAB could enhance the levels of total flavonoids and total polyphenols in FAS. Additionally, the beneficial components such as Delphinidin chloride, Diosmetin, Psoralidin, and Catechol significantly increased (p < 0.05). The colitis treatment experiment demonstrated that fermented AS could alleviate symptoms and improve the morphology of colitis in mice by enhancing antioxidant enzymes like CAT, T-SOD, and T-AOC. It also regulated the composition and abundance of intestinal flora species, such as Lactobacillus and Pseudogracilibacillus. The effectiveness of fermented AS was significantly superior to that of unfermented AS (p < 0.05). In conclusion, this study contributes to the application of lactic acid bacteria in AS fermentation and reveals the mechanism of fermentation AS for colitis.

Human breast milk isolated lactic acid bacteria: antimicrobial and immunomodulatory activity on the Galleria mellonella burn wound model.

Introduction: Managing burn injuries is a challenge in healthcare. Due to the alarming increase in antibiotic resistance, new prophylactic and therapeutic strategies are being sought. This study aimed to evaluate the potential of live Lactic Acid Bacteria for managing burn infections, using Galleria mellonella larvae as an alternative preclinical animal model and comparing the outcomes with a common antibiotic. Methods: The antimicrobial activity of LAB isolated from human breast milk was assessed in vitro against Pseudomonas aeruginosa ATCC 27853. Additionally, the immunomodulatory effects of LAB were evaluated in vivo using the G. mellonella burn wound infection model. Results and discussion: In vitro results demonstrated the antimicrobial activity of Lactic Acid Bacteria against P. aeruginosa. In vivo results show that their prophylactic treatment improves, statistically significant, larval survival and modulates the expression of immunity-related genes, Gallerimycin and Relish/NF-κB, strain-dependently. These findings lay the foundation and suggest a promising alternative for burn wound prevention and management, reducing the risk of antibiotic resistance, enhancing immune modulation, and validating the potential G. mellonella as a skin burn wound model.

Effects of gnotobiotic fermentation on global gene expression of germ-free vegetables.

Existing research has underscored the vital interplay between host organisms and their associated microbiomes, which affects health and function. In both plants and animals, host factors critically shape microbial communities and influence growth, health, and immunity. Post-harvest plants, such as those used in kimchi, a traditional Korean dish, offer a unique avenue for exploring host-microbe dynamics during fermentation. Despite the emphasis on lactic acid bacteria (LAB) in fermentation studies, the roles of host factors remain unclear. This study aimed to investigate the influence of these factors on plant transcriptomes during kimchi fermentation. We individually inoculated nine LAB strains into germ-free kimchi to generate LAB-mono-associated gnotobiotic kimchi and performed RNA-sequencing analysis for the host vegetables during fermentation. The transcriptomes of post-harvest vegetables in kimchi change over time, and microbes affect the transcriptome profiles of vegetables. Differentially expressed gene analyses revealed that microbes affected the temporal expression profiles of several genes in the plant transcriptomes in unique directions depending on the introduced LAB strains. Cluster analysis with other publicly available transcriptomes of post-harvest vegetables and fruits further revealed that the plant transcriptome is more profoundly influenced by the environment harboring the host than by host phylogeny. Our results bridge the gap in understanding the bidirectional relationship between host vegetables and microbes during food fermentation, illuminating the complex interplay between vegetable transcriptomes, fermentative microbes, and the fermentation process in food production. The different transcriptomic responses elicited by specific LAB strains suggest the possibility of microbial manipulation to achieve the desired fermentation outcomes.

Lactic acid bacteria isolated from women' breast milk and infants' faeces have appreciable immunogenic and probiotic potentials against diarrheagenic E. coli strains.

Diarrheal diseases remain the leading cause of high mortality among the infants, particularly in the developing countries; Probiotic intervention for diarrhea has been an ongoing novel approach to diarrheal prevention and treatment. This study aims to characterize immunogenic and probiotic properties of lactic acid bacteria (LAB) isolated from human breast milk and neonates' faeces. The LAB isolates from 16 mothers' breast milk and 13 infants' faeces were screened and identified by 16 S rRNA gene partial sequencing. Their antimicrobial activities against 5 strains of diarrheagenic Escherichia coli were tested. Organic acids production was quantified by HPLC, and antibiotic resistance pattern were determined by VITEK®. Autoaggregation, co-aggregation and hydrophobicity properties were assessed by UV spectrophotometry and immunomodulatory effect was determined in mouse model. Ninety-three LAB of five genera were identified. The most abundant species was Lactiplantibacillus plantarum with inhibition zones ranged from 8.0 to 25.0 ± 1 mm. Lacticaseibacillus rhamnosus A012 had 76.8 mg/mL lactic acid, (the highest concentration), was susceptible to all antibiotics tested. L. plantarum A011 and L. rhamnosus A012 were highly resistance to gastrointestinal conditions. L. rhamnosus A012 produced hydrophobicity of 25.01% (n-hexadecane), 15.4% (xylene) and its autoaggregation was 32.52%. L. rhamnosus A012 and L. plantarum A011 exert immunomodulatory effects on the cyclophosphamide-treated mice by upregulating anti-inflammatory cytokine and downregulating proinflammatory cytokines. Lactobacillus sp. demonstrated good probiotic and immunomodulatory properties. Further works are ongoing on the practical use of the strains.

Characterization of Lactic Acid Bacteria Isolated From Rotting Oranges and Use of Agropastoral Processing By-products as Carbon and Nitrogen Sources Alternative for Lactic Acid Production.

This study investigated the ability of lactic acid bacteria (LAB) isolated from oranges to use fish by-products (FB) and chicken by-products (CB) as nitrogen sources alternative to yeast extract for lactic acid (LA) production in a papaya by-product medium as a carbon source. Once the fermentation agents had been isolated, they were subjected to biochemical and molecular characterization. Inexpensive nitrogen sources, precisely CB and FB, were prepared, freeze-dried, and yield evaluated. Also, before to the fermentation experiments, the Total Kjehdahl Nitrogen (TKN) of these by-products and that of the yeast extract were determined. Then, three production media differing in terms of nitrogen source were formulated from these nitrogen sources. From the 22 LAB isolated from orange, two isolates of interest (NGO25 and NGO23) were obtained; all belonging to the Lactiplantibacillus plantarum species based on 16S rRNA gene sequencing. Furthermore, the production yield powder obtained after lyophilization of 1 L of CB and FB surpernatant were, respectively, 16.6 g and 12.933 g. The TKN of different nitrogen sources powder were 71.4 ± 0.000% DM (FB), 86.145 ± 0.001% DM (CB), and 87.5 ± 0.99% DM (yeast extract). The best kinetic parameters of LA production (LA (g/L): 31.945 ± 0.078; volumetric productivity (g/L.h): 1.331 ± 0.003; LA yield (mg/g) 63.89 ± 0.156; biomass (g/L) 7.925 ± 0.035; cell growth rate (g/L.h): 0.330 ± 0.001) were recorded by Lactiplantibacillus plantarum NGO25 after 24 h of fermentation. The latter data were obtained in the production medium containing CB as nitrogen sources. In addition, this production medium cost only $0.152 to formulate, compared to yeast extract which required $1.692 to formulate. Thus, freeze-dried CB can be used as an alternative to yeast extract in large-scale production of LA.

Effects of temperature and lactic acid Bacteria additives on the quality and microbial community of wilted alfalfa silage.

This study investigated the influence of different temperatures (35℃ High temperature and average indoor ambient temperature of 25℃) and lactic acid bacterial additives (Lactiplantibacillus plantarym, Lentilactobacillus buchneri, or a combination of Lactiplantibacillus plantarym and Lentilactobacillus buchneri) on the chemical composition, fermentation quality, and microbial community of alfalfa silage feed. After a 60-day ensiling period, a significant interaction between temperature and additives was observed, affecting the dry matter (DM), crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF) of the silage feed (p < 0.05). Temperature had a highly significant impact on the pH value of the silage feed (p < 0.0001). However, the effect of temperature on lactic acid, acetic acid, propionic acid, and butyric acid was not significant (p > 0.05), while the inoculation of additives had a significant effect on lactic acid, acetic acid, and butyric acid (p > 0.05). As for the dynamic changes of microbial community after silage, the addition of three kinds of bacteria increased the abundance of lactobacillus. Among all treatment groups, the treatment group using complex bacteria had the best fermentation effect, indicating that the effect of complex lactic acid bacteria was better than that of single bacteria in high temperature fermentation. In summary, this study explained the effects of different temperatures and lactic acid bacterial additives on alfalfa fermentation quality and microbial community, and improved our understanding of the mechanism of alfalfa related silage at high temperatures.

Microbiological quality and native lactic acid bacteria diversity of artisanal Mexican cheeses: A review.

This review aims to provide an overview of artisanal Mexican cheeses microbiota focused on microbiological quality and safety, as well as native Lactic acid Bacteria (LAB) diversity. For the search, key words of artisanal Mexican cheeses varieties was carried out through several online databases and original articles were screened and data about populations of indicator microorganisms, presence of pathogens, and native LAB identified were extracted. Several artisanal Mexican cheeses exceeded the permissible limit established in Mexican regulation (NOM-243-SSA1-2010) for indicator microorganisms, as well as in some types of cheese, the presence of pathogens was confirmed. However, other varieties of artisanal Mexican cheeses possess unique physicochemical characteristics, and during their manufacturing particular steps are used that contribute to ensuring their quality and safety. Additionally, strains able to control the growth of pathogenic and spoilage bacteria are part of the microbiota of some artisanal Mexican cheeses. About native LAB diversity, it is composed by species of Lactobacillus, Enterococcus, Streptococcus, Leuconostoc, Weisella, Lactococcus, Pediococus, Aerococus, Carnobacterium, Tetragenococus, among others genera. Otherwise, artisanal Mexican cheeses represent an important source of specific LAB with several approaches within human health because they showed potential for the development of functional foods, nutraceutical, and bioprotective cultures.

Insight into the autochthonous lactic acid bacteria as starter culture for improving the quality of Sichuan radish paocai: Changes in microbial diversity and metabolic profiles.

Paocai is a traditional Chinese fermented vegetable product popular in Asian countries. Recently, functional starters were used to control the fermentation process and improve the quality of paocai. In this study, three autochthonous lactic acid bacteria including Lactiplantibacillus plantarum LB6, Lactiplantibacillus pentosus LB3, and Weissella cibaria W51 were selected as starters and the effect of the starters on the fermentation of paocai was investigated. The results suggested that the inoculated fermentation led to a lower nitrite peak and more pronounced changes in pH and total titratable acid in the early stage of fermentation, compared with natural fermentation. Analysis of the flavor compounds indicated that the total content of volatile organic compounds of paocai through natural fermentation was significantly lower than that in inoculated fermentation. As for free amino acids, in the early stage of fermentation, the types and contents of free amino acids in the inoculated fermentation paocai were higher than those in the blank group. In the later stage of fermentation, the contents of amino acids representing umami and sweet tastes were also higher than those in the blank group. The bacterial community analysis showed that Lactobacillus and Lactococcus were the dominant bacteria in both inoculated fermentation and natural fermentation. Then, the correlations among physicochemical properties, microbial community and flavor compounds were revealed, and it was found that the dominant bacteria such as Lactococcus, Leuconostoc, Lactobacillus and Weissella displayed a considerable impact on the physical and chemical properties and flavor of paocai. In addition, the metabolic pathways involved in flavor formation and the abundance of related enzymes were elucidated. The abundance of enzymes involved in generating prephenic acid, 2-methylbutanoic acid, L-lactic acid, D-lactic acid, butanoic acid, etc., and in the pathway of producing flavor substances (His, Met, ethyl hexanoate, etc.) was up-regulated in the inoculated fermentation. Results presented in this study may provide a reference for the development of paocai starters and further guidance for the flavor improvement of Sichuan paocai.

Isolation and Identification of Lactic Acid Bacteria from Environmental Samples.

Isolation is the first and crucial step to study possible roles of lactic acid bacteria (LAB) in environment, especially in food fermentation. It is also important to use the organisms for further application. LABs are diverse bacterial group and have diverse growth characteristics. Culture condition of LAB is thus varied, and selection of a suitable culture medium is essential for the purposes. Identification is also important step, since certain desirable and undesirable characteristics are shared within species. Identification was classically carried out by phenotypic characteristics but is usually performed for DNA sequence-based approaches. 16S rRNA gene sequencing is generally used for the identification, and sequencing of housekeeping genes is used when needed. In addition, identification based on whole-genome sequence similarities was well established during the last decade. Here, we describe methods for isolation and identification of LAB briefly.

Introduction of Spontaneous Mutations Using Streptomycin as a Method for Lactic Acid Bacteria Breeding.

The chapter presents a technique for inducing spontaneous mutations using antibiotics that target microbial ribosomes and/or RNA polymerase, employed in bacterial breeding. In contrast to UV-based mutagenesis, this method allows control of the mutation sites, specifically targeting the rpsL gene. The outlined methodology introduces spontaneous mutations using streptomycin in Lacticaseibacillus rhamnosus GG ATCC 53103 (LGG), a widely studied lactic acid bacterium. Streptomycin has been shown to induce mutations in the rpsL gene, particularly altering lysine residues at position 56 or 101. It has also been reported to affect bacterial morphology and surface protein composition, thereby enhancing adhesion to human mucin.

Preparation and Structural Analysis of Lipoteichoic Acid on Cell Membranes Derived from Lactic Acid Bacteria.

Gram-positive bacteria, including lactic acid bacteria (LAB), possess lipoteichoic acid (LTA) on the cell surface. LTA is an amphiphilic molecule typically composed of hydrophilic glycerolphosphate polymer and hydrophobic anchor glycolipid moieties. It is involved in physiological properties of the cell surface and also plays roles in interactions with the host. Appropriate preparation procedures, such as extraction and purification, are required to clarify the structure-activity relationship. Structural diversity of LTA has been reported at the bacterial species and strain levels, and structural differences might affect interactions with the host. This chapter introduces techniques for preparation and structural analysis of LTA derived from LAB. It consists of four sections, covering butanol extraction, hydrophobic interaction chromatography, immunoblotting, and structural analysis. Technical notes containing supplemental information about the individual steps are also provided.

Yogurt Production.

Yogurt is a widely consumed dairy product that is obtained through the fermentation of lactic acid bacteria. During fermentation, these bacteria produce lactic acid, which lowers the pH and causes the coagulation of milk proteins. Metabolites of lactic acid bacteria, such as carbonyl compounds, nonvolatile or volatile acids, and exopolysaccharides, significantly affect the quality of yogurt. The production method also greatly influences yogurt characteristics. Yogurt can be produced in various forms, including solid, fluid, and soft-drinking types, depending on the manufacturing method employed. In addition, the raw materials, sterilization, homogenization, fermentation, and other conditions in each manufacturing method greatly affect yogurt's physical properties and flavor. This chapter summarizes common and modern methods of yogurt production.

Hydroxylation of Fatty Acids by Lactic Acid Bacteria.

Hydroxy fatty acids (HFAs) are fatty acids with hydroxyl functional groups attached to the main chain. HFAs are used in widely diverse industrial applications, healthy functional foods, artificial food flavorings, and alcoholic beverages. A lactic acid bacterium (LAB), Lactobacillus sakei, hydroxylates oleic acid. Furthermore, the hydroxyl fatty acid was identified by GC-MS as 10-hydroxystearic acid. The Lactobacillus sakei hydroxylated more than 90% of the oleic acid in the medium at 15 °C after 30-48 h. The hydroxyl enzyme needs a coenzyme for an electron donor as NADPH. The enzyme is useful for assay with monitoring NADPH concentration used an A340 device. The hydroxylate fatty acids are converted by LAB lactonize aroma lactone from commercial yeast strains, which can be detected directly by scent. Commercial beer brewing yeast T-58 produced the highest concentration of aroma lactone from hydroxyl fatty acids. Furthermore, the aroma lactone is identified by GC-MS as gamma-dodecalactone. The ratio of conversion is 87%. These results suggest that the lactonization conversion system is useful to hydroxylate fatty acids for alcoholic beverages.

Inhibition of Advanced Glycation End Products (AGEs) by Fermented Foods Using Lactic Acid Bacteria.

Glycation is an important nonenzymatic reaction between reducing sugars and amines. Advanced glycation end products (AGEs) accumulation in the human body is associated with secondary complications related to diabetes in hyperglycemic environments. These observations suggest that the inhibition of AGEs formation is important for preventing diabetes mellitus (DM) progression and the development of diabetes-related complications. Lactic acid bacteria (LAB) are probiotics commonly used in fermented foods and food additives. Therefore, it is necessary to identify starter strains of LAB to produce fermented food to decrease the risk of DM and its complications. This chapter introduces the protocols that are inhibition assay of fermented food using LAB on AGEs such as Nω-(carboxymethyl) arginine (CMA), Nε-(carboxymethyl) lysine (CML), and fluorescent AGEs.

Isolation of Lactic Acid Bacteria Eliminating Trimethylamine (TMA) for Application to Fishery Processing.

Fishy odor of fish flesh (meat) presents a severe problem for marine production. The main cause of fishy odor is trimethylamine (TMA), which increases during storage. It is produced from trimethylamine oxide (TMAO), an osmosis-regulating substance in fish cells that functions by a reduction reaction. Bacterial growth in fish meat increases TMA. Its odor reduces the commercial value of the meat. Technologies for its regulation and elimination are desired. This chapter presents a description of the use of lactic acid to eliminate TMA. The lactic acid is producible safely by bacteria during food processing using picric acid-toluene.A method of eliminating TMA was demonstrated using Lactobacillus plantarum H78. Furthermore, an assay method was explained for reducing TMA in fish meat by fermenting the H78 strain.

Lactic Acid Bacteria in the Human Oral Cavity: Assessing Metabolic Functions Relevant to Oral Health and Disease.

Many perceive lactic acid bacteria as beneficial for health. They are recognized for preventing abnormal fermentation and spoilage of ingested foods by producing lactic acid, which aids in gut acidification. Moreover, lactic acid bacteria are extensively employed in food science. In contrast, lactic acid bacteria in the oral cavity are often perceived as pathogenic factors contributing to the development of dental caries. As a consequence, substantial research has been conducted in oral and dental sciences to explore lactic acid bacteria and the oral microbiome. This research primarily involves analyzing bacterial flora, investigating metabolic activities such as acid production, and investigating metabolic regulation within the oral environment. The oral cavity serves as the gateway to the digestive tract and respiratory system, characterized by a constantly fluctuating environment that significantly impacts the metabolic activity of lactic acid bacteria. Hence, when investigating oral lactic acid bacteria, it is crucial to adopt research plans and methodologies that account for these metabolic environment changes. In this section, we present some of the methods employed in our study.

Assaying D-Alanine Racemase in Lactic Acid Bacteria Using NADH Oxidoreduction Enzymic System.

A notable characteristic of amino acids is their optical isomerism, existing as L-form and D-form. Proteins are composed exclusively of L-form amino acids. However, recently, it is reported that D-alanine is evaluated particularly highly in terms of sensory evaluation. D-body amino acids convert L-body amino acid proteolysis from a substrate such as foods during fermentation of lactic acid bacteria. This chapter presents a description of methods used for D-alanine racemase assays in the solution producing by lactic acid bacteria (LAB) using D-amino acid oxidase and lactic acid dehydrogenase via a NADH oxidoreduction system.

Biosorption of Histamine by Lactic Acid Bacteria for Detoxification.

Histamine accumulates in fish and fish products such as tuna, mackerel, skipjack, and bonito by work microorganisms. And it causes allergy reactions like IgE-mediated ones. Lactic acid bacteria (LAB) are known as one of the probiotic bacteria that indicate various health functionalities for humans. And some previous studies report that LAB can adsorb and excrete various toxic molecules. Here, this chapter introduces the methods to quantify the histamine-binding ability of LAB.