The prFMNH2-binding chaperone LpdD assists UbiD decarboxylase activation.

The UbiD enzyme family of prenylated flavin (prFMN)-dependent reversible decarboxylases is near ubiquitously present in microbes. For some UbiD family members, enzyme activation through prFMNH2 binding and subsequent oxidative maturation of the cofactor readily occurs, both in vivo in a heterologous host and through in vitro reconstitution. However, isolation of the active holo-enzyme has proven intractable for others, notably the canonical Escherichia coli UbiD. We show that E. coli heterologous expression of the small protein LpdD-associated with the UbiD-like gallate decarboxylase LpdC from Lactobacillus plantarum-unexpectedly leads to 3,4-dihydroxybenzoic acid decarboxylation whole-cell activity. This activity was shown to be linked to endogenous E. coli ubiD expression levels. The crystal structure of the purified LpdD reveals a dimeric protein with structural similarity to the eukaryotic heterodimeric proteasome assembly chaperone Pba3/4. Solution studies demonstrate that LpdD protein specifically binds to reduced prFMN species only. The addition of the LpdD-prFMNH2 complex supports reconstitution and activation of the purified E. coli apo-UbiD in vitro, leading to modest 3,4-dihydroxybenzoic acid decarboxylation. These observations suggest that LpdD acts as a prFMNH2-binding chaperone, enabling apo-UbiD activation through enhanced prFMNH2 incorporation and subsequent oxidative maturation. Hence, while a single highly conserved flavin prenyltransferase UbiX is found associated with UbiD enzymes, our observations suggest considerable diversity in UbiD maturation, ranging from robust autocatalytic to chaperone-mediated processes. Unlocking the full (de)carboxylation scope of the UbiD-enzyme family will thus require more than UbiX coexpression.

Comparative genomic analysis of Periweissella and the characterization of novel motile species.

The genus Periweissella was proposed as a novel genus in the Lactobacillaceae in 2022. However, the phylogenetic relationship between Periweissella and other heterofermentative lactobacilli, and the genetic and physiological properties of this genus remain unclear. This study aimed to determine the phylogenetic relationship between Periweissella and the two closest genera, Weissella and Furfurilactobacillus, by the phylogenetic analysis and calculation of (core gene) pairwise average amino acid identity. Targeted genomic analysis showed that fructose bisphosphate aldolase was only present in the genome of Pw. cryptocerci. Mannitol dehydrogenase was found in genomes of Pw. beninensis, Pw. fabaria, and Pw. fabalis. Untargeted genomic analysis identified the presence of flagellar genes in Periweissella but not in other closely related genera. Phenotypes related to carbohydrate fermentation and motility matched the genotypes. Motility genes were organized in a single operon and the proteins shared a high amino acid similarity in the genus Periweissella. The relatively low similarity of motility operons between Periweissella and other motile lactobacilli indicated the acquisition of motility by the ancestral species. Our findings facilitate the phylogenetic, genetic, and phenotypic understanding of the genus Periweissella.ImportanceThe genus Periweissella is a heterofermentative genus in the Lactobacillaceae which includes predominantly isolates from cocoa fermentations in tropical climates. Despite the relevance of the genus in food fermentations, genetic and physiological properties of the genus are poorly characterized and genome sequences became available only after 2020. This study characterized strains of the genus by functional genomic analysis, and by determination of metabolic and physiological traits. Phylogenetic analysis revealed that Periweissella is the evolutionary link between rod-shaped heterofermentative lactobacilli and the coccoid Leuconostoc clade with the genera Weissella and Furfurilactobacillus as closest relatives. Periweissella is the only heterofermentative genus in the Lactobacillaceae which comprises predominantly motile strains. The genomic, physiological, and metabolic characterization of Periweissella may facilitate the potential use of strains of the genus as starter culture in traditional or novel food fermentations.

Lactiplantibacillus plantarum J-15 reduced calcium oxalate kidney stones by regulating intestinal microbiota, metabolism, and inflammation in rats.

The prevention role of Lactiplantibacillus plantarum against the formation of kidney stones has been increasingly recognized; its mechanism, however, has mainly been focused on inhibiting the inflammation in the colon in the gastrointestinal (GI) system, and the intestinal metabolites from microflora have not been revealed fully with regarding to the stone formation. In this study, we investigated the effect of L. plantarum J-15 on kidney stone formation in renal calcium oxalate (CaOx) rats induced by ethylene glycol and monitored the changes of intestinal microflora and their metabolites detected by 16S rRNA sequencing and widely targeted analysis, followed by the evaluation of the intestinal barrier function and inflammation levels in the colon, blood and kidney. The results showed that L. plantarum J-15 effectively reduced renal crystallization and urinary oxalic acid. Ten microbial genera, including anti-inflammatory and SCFAs-related Faecalibaculum, were enriched in the J-15 treatment group. There are 136 metabolites from 11 categories significantly different in the J-15 supplementation group compared with CaOx model rats, most of which were enriched in the amino acid metabolic and secondary bile acid pathways. The expression of intestinal tight junction protein Occludin and the concentration of pro-inflammatory cytokines and prostaglandin were decreased in the intestine, which further reduced the translocated lipopolysaccharide and inflammation levels in the blood upon J-15 treatment. Thus, the inflammation and injury in the kidney might be alleviated by downregulating TLR4/NF-κB/COX-2 signaling pathway. It suggested that L. plantarum J-15 might reduce kidney stone formation by restoring intestinal microflora and metabolic disorder, protecting intestinal barrier function, and alleviating inflammation. This finding provides new insights into the therapies for renal stones.

Cupuassu juice fermentation by Lactiplantibacillus plantarum Lp62 produces an antioxidant enriched probiotic beverage.

The present work aimed to produce a cupuassu juice (Theobroma grandiflorum) fermented by the probiotic bacterium Lactiplantibacillus plantarum Lp62 and to analyze its antioxidant potential, antimicrobial activity, and resistance to biological barriers. The fermented beverage showed an increase in the content of phenolics, flavonoids, and antioxidant potential. The culture showed antagonistic activity against pathogens, but this result was not observed when the juice was tested. The probiotic strain remained viable under refrigeration, even in an acidified environment, and survived simulated gastrointestinal transit in vitro. L. plantarum Lp62 showed 30% adherence to HT-29 intestinal cells and proved to be safe in terms of antibiotic resistance and production of virulence factors. Fermentation increased the functional characteristics of cupuassu juice. This drink proved to be a good vehicle for the delivery of the probiotic bacteria L. plantarum Lp62.

Rye Dietary Fiber Components upon the Influence of Fermentation Inoculated with Probiotic Microorganisms.

Rye flour is used as the main ingredient of sourdough bread, which has technological and gastronomic benefits and increased nutritional value. The transformations observed during fermentation and baking may enable the conversion or degradation of rye dietary fiber carbohydrates built mainly of arabinoxylans, fructans, and ß-glucans. This study aimed to determine the dynamics of the changes in the contents of complex carbohydrates in sourdoughs inoculated with potential probiotic microorganisms as well as the polysaccharide composition of the resulting bread. Sourdoughs were inoculated with the potential probiotic microorganisms Saccharomyces boulardii, Lactiplantibacillus plantarum, Lacticaseibacillus rhamnosus, and Bacillus coagulans, and spontaneous fermentation was performed as a control. Samples of the sourdoughs after 24 and 48 h of fermentation and of bread obtained with these sourdoughs were analyzed for the content of individual dietary fiber components. The present study demonstrated that the treatments applied contributed to an increased total content of arabinoxylans in the breads, and the inoculation of the sourdoughs with the potential probiotic strains improved their solubility in water. The use of the S.boulardii strain may seem prospective as it allowed for the greatest reduction in fructans in the rye bread. Rye sourdough bread is an attractive source of dietary fiber and can be modified for different nutritional needs.

Ligilactobacillus agilis BKN88 possesses thermo-/acid-stable heteropolymeric flagellar filaments.

Many flagellated bacteria possess multiple flagellins, but the roles and the compositions of each flagellin are diverse and poorly understood. In Ligilactobacillus agilis BKN88, there are two active flagellin gene paralogues but their function and composition in its flagellar filaments have not been described. The aim of this study is to find the function and composition of the flagellins by employing mutant strains each of which expresses a single flagellin or a modified flagellin. Two single flagellin-expressing strains were both flagellated while the number of flagella per cell in the single flagellin-expressing derivatives was lower than that in the wild type. Nonetheless, these derivative strains were apparently equally motile as the wild type. This indicates that either flagellin is sufficient for cell motility. The immunological activity via Toll-like receptor 5 of the single flagellin-expressing strains or purified single flagellins was readily detectable but mostly variably weaker than that of the wild type. The flagellar filaments of wild type L. agilis BKN88 were more acid-/thermo-stable than those of single flagellin-expressing derivatives. Using a combination of immunoprecipitation and flagellin-specific staining, wild type BKN88 appeared to possess heteropolymeric flagellar filaments consisting of both flagellins and each flagellin appeared to be equally distributed throughout the filaments. The results of this study suggest that the two flagellins together form a more robust filament than either alone and are thus functionally complementary.

Histamine-forming ability of Lentilactobacillus parabuchneri in reduced salt Cheddar cheese.

Lentilactobacillus parabuchneri, a member of the non-starter microbiota in cheese, was recently associated with fast and effective histamine-formation ability, a safety issue. The present study was performed to investigate Lentilactobacillus parabuchneri KUH8, a histamine-producer (HP) in reduced-salt Cheddar cheese. Four cheeses were manufactured: 1) normal-salt (NS); 2) reduced-salt (RS); 3) normal-salt with HP (NS+HP); 4) reduced-salt with HP (RS+HP). Two replicates were produced with milk from the same batch, and the cheeses ripened at 10 and 15 °C. Cheeses were sampled immediately after manufacture and after 1, 3 and 6 months of ripening. Ultra-high-performance-liquid chromatography indicated that with the HP, histamine reached higher levels in reduced-salt cheeses (3.5-3.7% S/M) at 15 °C (86, 1112, 2149 and 3149 mg kg-1), compared to normal-salt cheeses (5.4-6.3% S/M) at 10 °C (78, 584, 593 and 1389 mg kg-1), at each respective cheese-sampling point. Higher salt-content reduced the growth rate of non-starter microbiota, but after six months the levels in all cheeses were similar, according to the ripening temperature: at 10 °C (8.05-8.30 log10 cfu g-1), and at 15 °C (6.00-6.94 log10 cfu g-1). A correlation between increased histamine levels, non-starter-cell development and pH was found. This study highlights the importance of normal-salt content and low-ripening temperature as measures to control histamine-formation and to improve safety in cheese.

Bioprotective extracts from Lactobacillus acidophilus CRL641 and Latilactobacillus curvatus CRL705 inhibit a spoilage exopolysaccharide producer in a refrigerated meat system.

The effect of bioprotective extracts (BEs) from Latilactobacillus curvatus CRL705 and Lactobacillus acidophilus CRL641 against Latilactobacillus sakei CRL1407 was evaluated in a refrigerated meat model system under vacuum and aerobic conditions at 4 and 10 °C. As shown by culturing, the BE-1 from L. acidophilus completely inhibited the spoilage strain, while that from Lat. Curvatus CRL705 (BE-2) and its combination with BE-1 exerted a bacteriostatic effect. The antimicrobial activity and exopolysaccharide production correlated with the efficacy of inhibitory treatment while final pH decrease was higher in control samples. When flow cytometry was applied, a lack of correlation with plate counting was found; counts under the detection limit for BE-1 at 21 and 28 days at 4 and 10 °C represented between 64.15 and 73.70% of dead cells. Thus, the concurrence of lactic acid bacteria as biocontrol agents and the use of more accurate tools to prevent the growth of deteriorating species will contribute to the extension of fresh meat shelf-life without quality loss.

Fermentation as a tool for increasing food security and nutritional quality of indigenous African leafy vegetables: the case of Cucurbita sp.

Sub-Saharan region is often characterized by food and nutrition insecurity especially "hidden hunger" which results from inadequate micronutrients in diets. African indigenous leafy vegetables (AILVs) can represent a valid food source of micronutrients, but they often go to waste resulting in post-harvest losses. In an attempt to prolong AILVs shelf-life while enhancing their nutritional quality, fermentation was studied from a microbiological and nutritional point of view. Pumpkin leaves (Cucurbita sp.) were spontaneously fermented using the submerged method with 3% NaCl and 3% sucrose. Controls were set up, consisting of leaves with no additions. During fermentation, samples of both treatments were taken at 0, 24, 48, 72 and 168 h to monitor pH and characterize the microbial population through culture-based and molecular-based analyses. Variations between fresh and treated leaves in B-group vitamins, carotenoids, polyphenols, and phytic acid were evaluated. Data revealed that the treatment with addition of NaCl and sucrose hindered the growth of undesired microorganisms; in controls, unwanted microorganisms dominated the bacterial community until 168 h, while in treated samples Lactobacillaceae predominated. Furthermore, the content in folate, ß-carotene and lutein increased in treated leaves compared to the fresh ones, while phytic acid diminished indicating an amelioration in the nutritional value of the final product. Thus, fermentation could help in preserving Cucurbita sp. leaves, avoiding contamination of spoilage microorganisms and enhancing the nutritional values.

Metatranscriptomics reveals the gene functions and metabolic properties of the major microbial community during Chinese Sichuan Paocai fermentation.

Chinese Sichuan Paocai (CSP) is one of the world's best-known fermented vegetables with a large presence in the Chinese market. The dynamic microbial community is the main contributor to Paocai fermentation. However, little is known about the ecological distribution and functional importance of these community members. In this study, metatranscriptomics was used to comprehensively explore the active microbial community members and key transcripts with significant functions in the Paocai fermentation process. Enterobacter, Leuconostoc, and Lactobacillus dominated the three-fermentation stages (Pre-, Mid- and Lat-), respectively. Carbon metabolism was the most abundant pathway. GH (glycoside hydrolase) and GT (lycosyl transferase) were the two most highly expressed carbohydrate-active enzymes. The most highly differentially expressed genes were grouped in the biosynthesis of amino acids, followed by glycolysis. Meta-pathways in the Sichuan Paocai fermentation ecosystem were reconstructed, Lactobacillaceae and Enterobacteriaceae were the two most important metabolic contributors. In addition, the nrfA and nirB were two genes referred to distinct nitrite reductase enzymes and 9 specialized genes, such as eclo, ron and ent were expressed to produce autoinducer 2 (AI-2) kinase in response to population density. The present study revealed functional enzymes and meta-pathways of the active microbial communities, which provide a deeper understanding of their contribution to CSP products.

3-Hydroxypropionic acid contributes to the antibacterial activity of glycerol metabolism by the food microbe Limosilactobacillus reuteri.

Strains of Limosilactobacillus reuteri are used as starter and bioprotective cultures and contribute to the preservation of food through the production of fermentation metabolites lactic and acetic acid, and of the antimicrobial reuterin. Reuterin consists of acrolein and 3-hydroxypropionaldehyde (3-HPA), which can be further metabolized to 1,3-propanediol and 3-hydroxypropionic acid (3-HP). While reuterin has been the focus of many investigations, the contribution of 3-HP to the antimicrobial activity of food related reuterin-producers is unknown. We show that the antibacterial activity of 3-HP was stronger at pH 4.8 compared to pH 5.5 and 6.6. Gram-positive bacteria were in general more resistant against 3-HP and propionic acid than Gram-negative indicator strains including common food pathogens, while spoilage yeast and molds were not inhibited by ≤ 640 mM 3-HP. The presence of acrolein decreased the minimal inhibitory activity of 3-HP against E. coli indicating synergistic antibacterial activity. 3-HP was formed during the growth of the reuterin-producers, and by resting cells of L. reuteri DSM 20016. Taken together, this study shows that food-related reuterin producers strains synthesize a second antibacterial compound, which might be of relevance when strains are added as starter or bioprotective cultures to food products.

The Carbohydrate Metabolism of Lactiplantibacillus plantarum.

Lactiplantibacillus plantarum has a strong carbohydrate utilization ability. This characteristic plays an important role in its gastrointestinal tract colonization and probiotic effects. L. plantarum LP-F1 presents a high carbohydrate utilization capacity. The genome analysis of 165 L. plantarum strains indicated the species has a plenty of carbohydrate metabolism genes, presenting a strain specificity. Furthermore, two-component systems (TCSs) analysis revealed that the species has more TCSs than other lactic acid bacteria, and the distribution of TCS also shows the strain specificity. In order to clarify the sugar metabolism mechanism under different carbohydrate fermentation conditions, the expressions of 27 carbohydrate metabolism genes, catabolite control protein A (CcpA) gene ccpA, and TCSs genes were analyzed by quantitative real-time PCR technology. The correlation analysis between the expressions of regulatory genes and sugar metabolism genes showed that some regulatory genes were correlated with most of the sugar metabolism genes, suggesting that some TCSs might be involved in the regulation of sugar metabolism.

Bioprospecting Antimicrobials from Lactiplantibacillus plantarum: Key Factors Underlying Its Probiotic Action.

Lactiplantibacillus plantarum (L. plantarum) is a well-studied and versatile species of lactobacilli. It is found in several niches, including human mucosal surfaces, and it is largely employed in the food industry and boasts a millenary tradition of safe use, sharing a long-lasting relationship with humans. L. plantarum is generally recognised as safe and exhibits a strong probiotic character, so that several strains are commercialised as health-promoting supplements and functional food products. For these reasons, L. plantarum represents a valuable model to gain insight into the nature and mechanisms of antimicrobials as key factors underlying the probiotic action of health-promoting microbes. Probiotic antimicrobials can inhibit the growth of pathogens in the gut ensuring the intestinal homeostasis and contributing to the host health. Furthermore, they may be attractive alternatives to conventional antibiotics, holding potential in several biomedical applications. The aim of this review is to investigate the most relevant papers published in the last ten years, bioprospecting the antimicrobial activity of characterised probiotic L. plantarum strains. Specifically, it focuses on the different chemical nature, the action spectra and the mechanisms underlying the bioactivity of their antibacterial and antiviral agents. Emerging trends in postbiotics, some in vivo applications of L. plantarum antimicrobials, including strengths and limitations of their therapeutic potential, are addressed and discussed.

Microbial Application to Improve Olive Mill Wastewater Phenolic Extracts.

Olive mill wastewater (OMW) contains valuable and interesting bioactive compounds, among which is hydroxytyrosol, which is characterized by a remarkable antioxidant activity. Due to the health claims related to olive polyphenols, the aim of this study was to obtain an extract from OMW with an increased level of hydroxytyrosol by means of microbial enzymatic activity. For this purpose, four commercial adsorbent resins were selected and tested. The beta-glucosidase and esterase activity of strains of Wickerhamomyces anomalus, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae were also investigated and compared to those of a commercial enzyme and an Aspergillus niger strain. The W. anomalus strain showed the best enzymatic performances. The SP207 resin showed the best efficiency in selective recovery of hydroxytyrosol, tyrosol, oleuropein, and total phenols. The bioconversion test of the OMW extract was assessed by using both culture broths and pellets of the tested strains. The results demonstrated that the pellets of W. anomalus and L. plantarum were the most effective in hydroxytyrosol increasing in phenolic extract. The interesting results suggest the possibility to study new formulations of OMW phenolic extracts with multifunctional microorganisms.

Effect of lactic fermentation on soy protein digestive pattern assessed by an in vitro dynamic gastrointestinal digestion model and the influence on human faecal microbiota.

BACKGROUND: The aim of this study was to investigate the effect of lactic fermentation on soy protein gastrointestinal digestive pattern and the influence of protein digesta on human faecal microbiota. Soymilk and soy yogurt were prepared in this study and a novel in vitro dynamic gastrointestinal model was employed to simulate gastric and duodenum digestions. Particle size, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and peptide content were monitored at the end of duodenum tract. RESULTS: Ingestion of soy yogurt allowed a rapid drop in pH from 7.0 to 5.0 at simulated duodenal digestion (0-30 min), and resulted in a loss in soluble protein content compared to that of soymilk. The electrophoretic pattern between soymilk and soy yogurt exerted distinctive differences at early stages of duodenal digestion (0-60 min) and resulted in different peptide contents (180 min). Soy yogurt duodenal digesta collected at 180 min (D180), by co-fermentation with human intestinal flora distribution, allowed a higher population in Bifidobacterium spp., Lactobacillus/Enterococcus spp. and Streptococcus/Lactococcus spp., whereas soy yogurt D30 resulted in lower population in Clostridium and Escherichia coli compared to samples co-fermented with soymilk digesta. CONCLUSION: The results demonstrated lactic fermentation of soy protein modulated human intestinal microflora and might relate to the different protein digestive behaviours. © 2020 Society of Chemical Industry.

Lactic acid production using cheese whey based medium in a stirred tank reactor by a ccpA mutant of Lacticaseibacillus casei.

In lactobacilli, CcpA is known to modulate the expression of genes involved in sugar metabolism, stress response and aerobic adaptation. This study aimed to evaluate a ccpA mutant of Lacticaseibacillus casei BL23 to increase lactic acid production using cheese whey. The ccpA derivative (BL71) showed better growth than the L. casei wild-type in the whey medium. In a stirred tank reactor, at 48 h, lactate production by BL71 was eightfold higher than that by BL23. In batch fermentations, the final values reached were 44.23 g L-1 for BL71 and 27.58 g L-1 for BL23. Due to a decrease in the delay of lactate production in the mutant, lactate productivity increased from 0.17 g (L.h)-1 with BL23 to 0.80 g (L.h)-1 with BL71. We found that CcpA would play additional roles in nitrogen metabolism by the regulation of the proteolytic system. BL71 displayed higher activity of the PepX, PepQ and PrtP enzymes than BL23. Analysis of prtP expression confirmed this deregulation in BL71. Promoter analysis of the prtP gene revealed CcpA binding sites with high identity to the cre consensus sequence and the interaction of CcpA with this promoter was confirmed in vitro. We postulate that deregulation of the proteolytic system in BL71 allows a better exploitation of nitrogen resources in cheese whey, resulting in enhanced fermentation capacity. Therefore, the ccpA gene could be a good target for future technological developments aimed at effective and inexpensive lactate production from dairy industrial wastes.

Inactivation kinetics of beer spoilage bacteria (Lactobacillus brevis, Lactobacillus casei, and Pediococcus damnosus) during acid washing of brewing yeast.

This work aimed to estimate the inactivation kinetic parameters of four potential beer spoilage bacteria (Lactobacillus brevis DSM 6235, Lactobacillus casei ATCC 334, Pediococcus damnosus DSM 20289 and Pediococcus damnosus ATCC 29358) inoculated in brewing yeast submitted to acid washing with purposes of yeast recycle. The experiments were conducted at 4 °C in solutions with pH 1.5, pH 2, and pH 3 adjusted employing 85% phosphoric acid. The acid washing treatment of brewing yeasts in the most common pH used (pH 2.0) demanded almost 50 min for the first decimal reduction (δ) of L. brevis DSM 6235. Sensible strains to acid washing such as P. damnosus DSM 20289 demanded almost 70 min for 4 log reductions to be achieved. On the other hand, pH reduction of the acid washing from 2.0 to 1.5 allowed 4 log reduction of L. brevis DSM 6235) to be obtained in less than 50 min, without ruining brewer's yeast viability. Acid washing in pH 1.5 is a viable method for the inactivation of bacterial contaminants of brewing yeasts. Recycling of brewing yeasts through this approach may contribute to a more sustainable and environmental-friendly industry.

Comparison of backslopping and two-stage fermentation methods for koumiss powder production based on chemical composition and nutritional properties.

BACKGROUND: Koumiss is a traditional fermented beverage made from mare's milk. The traditional backslopping method for koumiss production has shortcomings in terms of microbiological diversity and nutritional characteristics. In this study, a two-stage fermentation method was established to scale up the production of koumiss powder. The chemical composition and nutritional properties of a novel two-stage fermentation koumiss powder (TKP) were compared with backslopping koumiss powder (BKP). RESULTS: The TKP exhibited important nutritional and functional properties, including a high percentage of essential amino acids, and high polyunsaturated fatty acid, vitamin, and mineral content. The essential amino acid content of TKP was not significantly different from that of BKP. The oleic acid, linoleic acid, linolenic acid, and water-soluble vitamin content of TKP was higher than that of BKP. The Ca:P ratio of TKP was also close to the optimal Ca:P ratio in humans. CONCLUSION: The novel method could be applied for the scaled-up production of koumiss powder with similar nutritional properties to traditional backslopping koumiss powder. The successful production of koumiss powder could also promote the development of the koumiss industry. © 2019 Society of Chemical Industry.

Sharpea azabuensis: a ruminal bacterium that produces trans-11 intermediates from linoleic and linolenic acid.

To investigate the metabolism of 18:2n-6 and 18:3n-3 by pure cultures of Sharpea azabuensis, two different strains (RL 1 and ST18) were each incubated in the presence of 40 µg ml-1 18:2n-6 or 18:3n-3. Pure cultures of Butyrivibriofibrisolvens D1 and Butyrivibrio proteoclasticus P18 were included as control treatments. Similar to the metabolism of B. fibrisolvens, both S. azabuensis strains converted 18:2n-6 or 18:3n-3 to cis-9, trans-11 CLA or cis-9, trans-11, cis-15 CLnA, after which it was further reduced to trans-11 18:1 or trans-11, cis-15 18:2, respectively. B. proteoclasticus additionally reduced trans-11 18:1 to 18:0. Trans-11, cis-15 18:2 was also further metabolized by B. proteoclasticus, although trans-11 18:1 did not accumulate, and only minor amounts of 18:0 were formed. The time frame of 18:2n-6 and 18:3n-3 biohydrogenation by S. azabuensis was comparable with B. fibrisolvens, indicating that S. azabuensis and B. fibrisolvens might be alternative biohydrogenators of 18:2n-6 and 18:3n-3 in the rumen.

Wheat Germ Supplementation Increases Lactobacillaceae and Promotes an Anti-inflammatory Gut Milieu in C57BL/6 Mice Fed a High-Fat, High-Sucrose Diet.

BACKGROUND: A link between high-fat diet consumption and obesity-related diseases is the disruption of the gut bacterial population, which promotes local and systemic inflammation. Wheat germ (WG) is rich in bioactive components with antioxidant and anti-inflammatory properties. OBJECTIVE: The aim of this study was to investigate the effects of WG supplementation in modulating the gut bacterial population and local and systemic inflammatory markers of mice fed a high-fat, high-sucrose (HFS) diet. METHODS: Six-week-old male C57BL/6 mice were randomly assigned to 4 groups (n = 12/group) and fed a control (C; 10% kcal fat, 10% kcal sucrose) or HFS (60% kcal fat, 20% kcal sucrose) diet with or without 10% WG (wt:wt) for 12 wk. Cecal bacteria was assessed via 16S rDNA sequencing, fecal short-chain fatty acids by GC, small intestinal CD4+ lymphocytes using flow cytometry, and gut antimicrobial peptide genes and inflammatory markers by quantitative polymerase chain reaction. Statistical analyses included Kruskal-Wallis/Dunn's test and 2-factor ANOVA using HFS and WG as factors. RESULTS: There was a 4-fold increase (P = 0.007) in the beneficial bacterial family, Lactobacillaceae, in the HFS + WG compared with the HFS group. Fecal propionic and n-butyric acids were elevated at least 2-fold in C + WG compared with the other groups (P < 0.0001). WG tended to increase (≥7%; P-trend = 0.12) small intestinal regulatory T cell:Th17 ratio, indicating a potential to induce an anti-inflammatory gut environment. WG elevated (≥35%) ileal gene expression of the anti-inflammatory cytokine Il10 compared to the unsupplemented groups (P = 0.038). Ileal gene expression of the antimicrobial peptides Reg3b and Reg3g was upregulated (≥95%) in the HFS + WG compared with other groups (P ≤ 0.040). WG reduced serum concentrations of the pro-inflammatory cytokines, interleukin (IL)-1B, IL-6, interferon-γ, and tumor necrosis factor-α (≥17%; P ≤ 0.012). CONCLUSIONS: WG selectively increased gut Lactobacillaceae, upregulated ileal antimicrobial peptides, and attenuated circulating pro-inflammatory cytokines of C57BL/6 mice fed a HFS diet. These changes may be vital in preventing HFS diet-induced comorbidities.