Propionibacterium Freudenreichii Research Articles

Matching excellence: Oxford Nanopore Technologies' rise to parity with Pacific Biosciences in genome reconstruction of non-model bacterium with high G+C content.

The reconstruction of complete bacterial genomes is essential for microbial research, offering insights into genetic content, ontology and regulation. While Pacific Biosciences (PacBio) provides high-quality genomes, its cost remains a limitation. Oxford Nanopore Technologies (ONT) offers long reads at a lower cost, yet its error rate raises scepticism. Recent ONT advancements, such as new Flow cells (R10.4.1), chemistry (V14) and duplex mode, improve data quality. Our study compares ONT with PacBio and Illumina, including hybrid data. We used Propionibacterium freudenreichii, a bacterium with a genome known for being difficult to reconstruct. By combining data from ONT's Native Barcoding and a custom-developed BARSEQ method, we achieved high-quality, near-perfect genome assemblies. Our findings demonstrate, for the first time, that the combination of nanopore-only long-native with shorter PCR DNA reads (~3 kb) results in high-quality genome reconstruction, comparable to hybrid data assembly from two sequencing platforms. This endorses ONT as a cost-effective, stand-alone strategy for bacterial genome reconstruction. Additionally, we compared methylated motif detection between PacBio and ONT R10.4.1 data, showing that results comparable to PacBio are achievable using ONT, especially when utilizing the advanced Nanomotif tool.

Anti‐mold properties of fermented apple juice in bread

AbstractBackground and ObjectivesBread may be spoiled by mold, which results in major economic losses. The aim of this study was to optimize the anti‐mold properties of apple juice fermented with Propionibacterium freudenreichii to get high levels of propionic acid with lesser amounts of acetic acid, which impairs bread taste.FindingsAfter fermentation for 7 days at 35°C, apple juice (half‐diluted to about 4% fermentable sugars; with 1% yeast extract; pH adjusted to 6.0) contained 1.0% propionic acid and little amounts of acetic acid (about 0.35%).ConclusionsIn bread, the anti‐mold activity of fermented apple juice (5%, fb) was similar to that of apple cider vinegar (2.44%, fb).Significance and NoveltyFermented apple juice is a novel mold inhibitor for bread that may replace apple cider vinegar and part of calcium propionate.

Propionibacterium freudenreichii MJ2-derived extracellular vesicles inhibit RANKL-induced osteoclastogenesis and improve collagen-induced rheumatoid arthritis

Rheumatoid arthritis causes excessive bone loss by stimulating osteoclast differentiation. Extracellular vesicles are valuable disease markers, conveyors of distant cell-to-cell communication, and carriers for drug delivery. The aim of this study was to investigate the anti-osteoclastogenic effects of extracellular vesicles derived from dairy Propionibacterium freudenreichii MJ2 (PFEVs) and the improvement effect of PFEVs on collagen-induced arthritis (CIA) animal model. PFEVs were observed by scanning electron microscopy, transmission electron microscopy, nanoparticle tracking analysis, and LC–MS/MS. The inhibitory activity of PFEVs against receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation was investigated in RAW 264.7 cells. PFEVs significantly decreased the expression levels of genes and proteins related to osteoclast differentiation. PFEVs decreased RANK–RANKL binding. In a CIA mouse model, PFEVs treatment significantly reduced arthritis scores and collagen-specific immunoglobulins. PFEVs treatment also reduced pro-inflammatory cytokines and increased anti-inflammatory cytokines. The anti-inflammatory effects were confirmed by H&E staining, and PFEVs treatment inhibited osteoclastogenesis in the CIA mouse model. In conclusion, PFEVs inhibited osteoclast differentiation by inhibiting RANK–RANKL signaling, thereby decreasing the expression of osteoclast differentiation-related genes. PFEVs also improved collagen-induced arthritis by inhibiting inflammation and osteoclastogenesis.

A multispecies bacterial-based direct-fed microbial alleviates Salmonella invasion and supports in vitro epithelial integrity.

Managing bacterial infections is of great importance in livestock production, particularly those caused by Salmonella enterica serovars Typhimurium or Dublin, which can impact both animal health and performance, as well as human food safety. Direct-fed microbials (DFM) can support gastrointestinal function and alleviate the potential negative effects of bacterial infections. In the present study, the capacity of a multispecies bacterial-based DFM containing Ligilactobacillus (formerly Lactobacillus) animalis 506, Propionibacterium freudenreichii 507, Bacillus licheniformis 809, and B. subtilis 597 to reduce S. Typhimurium ATCC14028 invasion was investigated using a co-incubation model with the HT29-MTX-E12 cell line (experiment 1). Next, a possible antagonistic effect of the DFM against S. Dublin ATCC 41286 was evaluated using an in vitro agar well diffusion method following a co-incubation of 48 h (experiment 2). At last, a series of experiments were performed to evaluate how different doses (6.25 × 106, 2.50 × 107, or 1.00 × 108 CFU/well) of the DFM would support the integrity of intestinal epithelial cells challenged or not with S. Typhimurium ATCC14028 or hydrogen peroxide under a transepithelial electrical resistance (TEER) assay with Caco-2 cells (experiments 3 and 4). In experiment 1, BDP significantly (P < 0.001) reduced by 90.8% the invasion of S. Typhimurium into HT29-MTX-E12 cells, whereas viability of the potentially harmful bacteria was reduced by 21.0% (P < 0.0001). In experiment 2, the antagonistic properties of BDP towards S. Dublin were confirmed by the detection of a clear inhibition zone (size = 8.6 mm). Lastly, without challenge, the lowest dose of the DFM (6.25 × 106 CFU) provided the greatest support to the cells (treatment × hour; P < 0.0001). However, when the cells were challenged with S. Typhimurium, all doses alleviated the loss of integrity caused by the pathogen (treatment × hour; P < 0.0001). In cells challenged with hydrogen peroxide, the greater dose (1.00 × 108 CFU) supported the cells for a longer period of time (treatment × hour; P < 0.0001). These in vitro findings set the stage for exploring the potential benefits of using a novel DFM as a promising tool and strategy to mitigate S. enterica infections in ruminants and improve animal health, food safety, and public health. Further, in vivo confirmation needs to be developed to validate these preliminary in vitro results.

Investigation of Feedlot-level Use of a Direct-fed Microbial on Fecal Shedding of E. coli O157:H7

Our objectives were to determine whether the feedlot-level use of a direct-fed microbial (DFM; Lactobacillus animalis LA51 and Propionibacterium freudenreichii PF24; Bovamine Defend®, 2 × 109 CFU/g) was associated with fecal prevalence and concentration of E. coli O157:H7, and determine pen- and feedlot-level risk factors associated with fecal E. coli O157:H7 prevalence in cattle pens from commercial feedlot operations. Twenty commercial feedlots in Nebraska, ten that included DFM (DFM) and ten that did not (no-DFM), were sampled during the summer of 2017. In each sampling month, 22 pen-floor fecal samples were collected from three pens in each feedlot. Samples were subjected to cultural and molecular procedures for the detection of E. coli O157:H7 (immunomagnetic separation, plating on selective media, followed by PCR confirmation) and spiral plating for quantification. A total of 1,320 samples from 180 pens of finishing cattle belonging to 20 feedlots, which were sampled three times throughout a 12-week period, were processed and tested. Across all feedlots and sampling months, the mean within-pen prevalence was 13.5% (95% CI = 2.6–47.4%). The association between DFM status and the within-pen prevalence of E. coli O157:H7 depended significantly (p < 0.05) on the sampling month. The second sampling month between late July and mid-August corresponded to the highest within-pen prevalence estimates reported in this study, with no-DFM pens having a higher prevalence than DFM pens. After accounting for the DFM status, and based on multivariable analyses, sampling month, average pen body weight, and weather conditions were significantly associated with the within-pen fecal prevalence of E. coli O157:H7. Collectively, these findings demonstrate that the use of a DFM containing Lactobacillus animalis LA51 and Propionibacterium freudenreichii PF26 in feedlots showed potential in reducing fecal E. coli O157:H7 prevalence in cattle during times when prevalence peaks.

Aerobic adaptation and metabolic dynamics of Propionibacterium freudenreichii DSM 20271: insights from comparative transcriptomics and surfaceome analysis.

The study of the response of Propionibacterium freudenreichii to aerobic growth is crucial for understanding how this bacterium adapts to different environments and produces essential compounds like vitamin B12. By investigating its physiological changes under aerobic conditions, we can gain insights into its metabolic adjustments and potential for enhanced growth. These findings not only deepen our understanding of P. freudenreichii's responses to oxygen availability but also offer valuable information for optimizing its growth in industrial applications. This research sheds light on the adaptive mechanisms of this bacterium, providing a foundation for further exploration and potential applications in various fields.

Microbial chaperonin 60 inhibits osteoclast differentiation by interfering with RANK/RANKL binding and overexpression of lipocalin2

Osteoclasts play a crucial role in bone destruction in rheumatoid arthritis (RA). This study aimed to investigate the inhibitory effects of chaperonin 60 (CPN60), identified in the surface proteins of Propionibacterium freudenreichii MJ2, on receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation, and elucidate the underlying mechanisms. Treatment with CPN60 inhibited RANKL-induced osteoclast differentiation by decreasing the expression of osteoclast differentiation-related genes and proteins. CPN60 interfered with the binding of RANKL to RANK, as elucidated using surface plasmon resonance (SPR) and immunofluorescence. In silico molecular docking analysis further supported the interference of CPN60 with the binding of RANKL and RANK. CPN60 suppressed the expression of molecules linked to the calcium-dependent pathway in RANKL-induced osteoclast differentiation at both mRNA and protein levels. Microarray analysis showed elevated expression of lipocalin 2 (Lcn2), which was closely linked to the inhibition of osteoclast differentiation in CPN60-treated RAW 264.7 cells. Inhibition of Lcn2 decreased the inhibitory effect of CPN60 on osteoclast differentiation, indicating that increased expression of Lcn2 by CPN60 contributes to the inhibition of osteoclastogenesis. In addition, CPN60 treatment alleviated arthritis symptoms in collagen-induced arthritis mice by reducing the generation of collagen-specific antibodies and inhibiting osteoclast differentiation. In conclusion, CPN60 of P. freudenreichii MJ2 interfered with RANKL–RANK binding, reduced the expression of genes and proteins related to osteoclast differentiation and upregulated Lcn2 expression, thereby inhibiting RANKL-induced osteoclast differentiation, which might contribute to ameliorate collagen-induced arthritis.

Exploring a novel direct-fed microbial for ruminants to mitigate in vitro Salmonella invasion and infection

Abstract Managing bacterial infections is of great importance in cattle production, particularly those caused by Salmonella enterica serovars Typhimurium or Dublin, which can impact both animal health and human food safety. Direct-fed microbial (DFM) can support gastrointestinal health and alleviate potential bacterial infections. In the present study, the capacity of a bacteria-based DFM product [BOVAMINE DEFEND Plus; BDP; Novonesis, Denmark (BDP)] containing Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus licheniformis, and B. subtilis to reduce Salmonella Typhimurium ATCC14028 invasion was investigated using the HT29-MTX cell line (Exp. 1) and also to evaluate the possible antagonistic effect against S. Dublin using an in vitro agar well diffusion method (Exp. 2). Briefly, in Exp. 1, after a 30 min pre-incubation with 1.0 ×108 CFU of BDP and/or 2.5 × 106 CFU of S. Typhimurium were added to each transwell. The quantity of S. Typhimurium invading the cells was measured after 90 min of co-incubation using CFU counts. As a control, the viability of the total S. Typhimurium was assessed to confirm the mode of action of BDP targets the pathogen invasion properties. In Exp. 2, the antagonistic efficacy of BDP against S. Dublin CHCC41286, isolated from cattle, was tested by adding BDP to wells in CLED agar plates casted with the pathogen above, zones of inhibition measured after 48 h incubation. In Exp. 1, BDP significantly (P &amp;lt; 0.05) reduced by 80.6 % the invasion of S. typhimurium into HT-29-MTX cells as well as from 16 % the viability of the pathogen. The antagonistic properties of BDP towards S. Dublin were confirmed by clear inhibition zones (8.6 mm). These two in vitro findings set the stage for exploring the potential benefits of using a novel DFM as a promising tool and strategy to mitigate Salmonella infections in ruminants and improve animal health, food safety, and public health. Further in vivo confirmation needs to be developed to validate these preliminary in vitro results.

123 Effects of a Saccharomyces cerevisiae based direct-fed microbial on performance, health, and rumen bacterial community of newly weaned beef steers during a 56-d receiving period

Abstract We examined the effects of a blend of Saccharomyces cerevisiae, multiple live probiotic bacteria and their fermentation products on performance, health, and the ruminal bacterial community of newly weaned beef steers during a 56-d receiving period. Newly weaned Angus crossbred steers [n = 40; 221 ± 25.6 kg body weight (BW); 180 ± 17 d of age] were stratified by BW into four pens (10 steers per pen) such that each pen had a similar average BW at the beginning of the experiment. The pens were randomly assigned to receive a corn silage basal diet (CON; n = 20) or the basal diet supplemented with 9 g·steer⁻¹·d⁻¹of PRO feed additive (PRO; n = 20). The PRO additive is a blend of Saccharomyces cerevisiae and the fermentation products of Enterococcus faecium, Bacillus licheniformis, Bacillus subtilis, Lactobacillus animalis, Propionibacterium freudenreichii. The DMI and water consumed were monitored using the GrowSafe intake nodes and custom flow meters, respectively. Body weights were recorded weekly to calculate average daily gain (ADG). Before morning feeding, 10 mL of blood was taken from each steer on d 0 to 7, and thereafter weekly for analyses of immune cells, plasma glucose and NEFAs. On d 56, rumen fluid samples (200 mL each) were collected from all the steers for microbiome analysis. Over the 56-d receiving period, the supplemental PRO had no effects on DMI, water intake, or ADG. However, compared with CON, beef steers fed supplemental PRO tended to have greater ADG (P = 0.08) and BW (P = 0.07) during the first 14 d of the study. There was a treatment × day interaction (P ≤ 0.05) for WBC, neutrophils and monocytes over the 56 d such that beef steers fed supplemental PRO had decreased blood concentrations on certain days during the first 7 d after weaning, indicating reduced inflammation or stress response. The results of the rumen microbiome analysis revealed that the relative abundance of complex fiber degrading or obligate proton-reducing bacterial genera such as Bacteroides, Ruminococcus gauvreauii group, Desulfovibrio, Syntrophococcus, and Acetitomaculum were greater (P ≤ 0.05) in beef steers fed supplemental PRO compared with CON. This study demonstrated that dietary supplementation of PRO improved the growth performance, reduced stress or inflammatory response during the initial days after weaning and altered the ruminal bacterial community towards increased relative abundance of bacterial genera associated with improved rumen function.

Ensuring Tree Protection, Growth and Sustainability by Microbial Isolates

Antimicrobial properties of the new strains of micro-organisms isolated from natural sources of various ecological niches in the Moscow region and the Republic of Tatarstan were studied. Antifungal activity of isolates was detected in a test culture of toxin-producing microscopic fungi that can cause animal and plant diseases: Aspergillus flavus, Candida albicans, Fusarium oxysporum and Penicillium spp. Of the 46 studied micro-organisms of genera Bacillus, Lactobacillus, Lactococcus and Streptomyces isolates, there are four strains (Bacillus subtilis, Lactobacillus plantarum, Propionibacterium freudenreichii and Streptomyces spp.) that showed an ability to produce biologically active metabolites with a pronounced antimicrobial potential against phytopathogenic fungi metabolites. Based on the selected four strains, a Bacterial product LRV composition has been created. Scots pine, pedunculate oak and small-leaved linden seedlings with single and double foliar treatment and Bacterial product LRV at a concentration of 10 mL/L led to an increase in the growth of the aboveground part by 31.8, 51.9 and 25.4%, respectively, and the underground part by 25.0, 37.2 and 25.7%, respectively, compared to the control. The weight of seedlings at the end of the study exceeded the control variant by an average of 26.0, 44.0 and 78.0%, respectively. Plant protection Bacterial product LRV use did not have a significant effect on the group of molds that caused the powdery mildew and Schütte disease damage to trees. The Biological product LRV provided plant protection from fungal diseases caused by Lophodermium pinastri Chev. and Microsphaera alphitoides.

Feed additives supplementation: a potential strategy to ameliorate heat stress in sheep

Abstract Given a significant climate-flexible and socio-economic role in developing nations, environmental heat stress imposes a major financial impact on sheep production systems globally endangering their production, reproduction, and growth. In this regard, the adverse effects of heat stress on sheep production systems have to be addressed through adoption of effective heat alleviation measures like animal management, nutritional management and genetic interventions of which the nutritional interventions seems to be the most cost effective way to alleviate heat stress. Nutritional manipulation for heat stress alleviation in sheep involves the use of antioxidant supplements (Vitamin B; Vitamin E and Selenium; Selenium; Zinc sulphate and folic acid; Vitamin C, Vitamin E, Selenium and Zinc; Naringin; Opuntia ficus-indica f. inermis; Açai oil and Brown seaweed like Ascophyllum nodosum and Sargassum latifolium). Further, electrolyte supplements (Dietary Electrolyte Balance (DEB); Sodium bicarbonate and potassium bicarbonate; Sodium hydroxide) have a beneficial effect on thermal responses, respiratory activities, gas exchange parameters, rumen fermentation, blood buffering capacity and acid-base balance. The mineral mixture supplements (Mineral blocks; Mineral mixture and antioxidants; Chromium; Zinc) play a crucial role in increasing the efficiency of antioxidant defence system, immunity-related parameters, production, reproduction, feed digestibility and insulin sensitivity. Probiotic supplements (Lactobacillus acidophilus, Saccharomyces cervisiae, Propionibacterium freudenreichii, Lactobacillus casei, Enterococcus faecium, Lactobacillus lactis, Bacillus subtilis, Propionibacterium freudenreichii, Pediococcus cerevisiae, Megaspha eraelsdenii, Bacillus licheniformis, Aspergillus oryzae, Schizochytrium limacinum, Trichoderma reesei and Saccharomyces cerevisiae) improve lactational performance, dietary energy utilization and productivity. The probiotics (live Saccharomyces cerevisiae) and prebiotics (mannan oligosaccharide plus b-glucans) used in heat stress alleviation improve dietary energy utilisation. Furthermore, the vital role of herbal supplements (Rosemary, Cinnamon, Turmeric, Clove, Naringin, Chestnut tannins, Giloy stem powder, Curcumin, Rocket oil (watercress oil), Flaxseed, Cornus, Oregano, Thyme, Chamomile flowers, Moringa oleifera, Betaine) has been highlighted to promote feed intake, antioxidant status, growth performance, feed utilization, reproductive performance and immune response. Effective adoption of nutritional strategies can thus ensure sustainable sheep production in this changing climate scenario.

The use of propionic and lactic acid bacteria to produce cobalamin and folate in injera, an Ethiopian cereal-based fermented food

Like in many developing countries, the traditional Ethiopian diet relies mainly on starchy staple foods and often lacks sufficient animal-sourced foods which are crucial for cobalamin intake. Furthermore, the concentration of folate in traditionally prepared injera, an Ethiopian cereal-based fermented staple food, is highly variable and injera contains biologically inactive corrinoids. This study aimed to improve the cobalamin and folate content of injera by using cobalamin-producing Propionibacterium freudenreichii and folate-producing Lactiplantibacillus plantarum strains, both individually and combined. Since injera is fermented using backslopping, we also assessed the ability of these strains to produce cobalamin and folate consistently across successive fermentation batches. Changes in the microbial ecosystem were monitored using real-time PCR. The theoretical contribution of the injera prepared using the selected strains to the cobalamin and folate intake of children and women of reproductive age was also calculated. Results showed that using the selected bacterial strains individually increased cobalamin (up to 19.2 μg/100 g of dry matter) and folate (up to 180.2 μg/100 g of dry matter) levels in the injera dough over several backslopping fermentation batches. Regular consumption of the injera with enhanced vitamin content produced using each strain alone would be capable of fulfilling the entire recommended nutrient intake for cobalamin and up to 29 % of the recommended intake for folate for children and women of reproductive age. However, when the strains were used together, the production of both vitamins was reduced. The presence of certain common endogenous bacterial species and genera exhibited significant variability, highlighting the complex response of the native microbiota to the different inoculation strategies employed. Future experiments should consider selecting a microbial consortium comprising non-competing microorganisms to ensure the simultaneous production of cobalamin and folate in fermented foods.

In situ fortification of cereal by-products with vitamin B12: An eco-sustainable approach for food fortification

Vitamin B12 deficiency poses significant health risks, especially among populations with limited access to animal-based foods. This study explores the utilisation of cereal bran by-products, wheat (WB) and oat bran (OB), as substrates for in situ vitamin B12 fortification through solid-state fermentation (SSF) using Propionibacterium freudenreichii. The impact of various precursors addition, including riboflavin, cobalt, nicotinamide and DMBI on vitamin B12 production, along with changes in microbial growth, chemical profiles, and vitamin B12 yields during fermentation was evaluated. Results showed that WB and OB possess favourable constituents for microbial growth and vitamin B12 synthesis. The substrates supplemented with riboflavin, cobalt, and DMBI demonstrated enhanced B12 production. In addition, pH levels are essential in microbial viability and cobalamin biosynthesis. Monosaccharides and organic acids play a crucial role, with maltose showing a strong positive association with B12 production in OB, while in WB, citric acid exhibits significant correlations with various factors.

In situ fortification of protein-enriched brewer's spent grain with vitamin B12 by fermentation with Priestia megaterium and Propionibacteriumfreudenreichii

Meat is an important protein source in the human diet; however, the consumption of animal proteins needs to be reduced. Plant-based foods naturally lack cobalamin, vitamin B12 (VitB12), an essential vitamin in the human diet. Brewer's spent grain (BSG) is a side stream available in large volumes that was chosen as a substrate for food-grade VitB12-producing microorganisms.VitB12-producing microorganisms were identified in the scientific literature and selected to study in situ production of VitB12 in a protein-enriched (53%) BSG. A first screening was performed using Propionibacterium freudenreichii and Priestia megaterium (formerly known as Bacillus megaterium) strains. Further optimisations were exploited to increase the BSG content further and resulted in levels of up to 21 μg VitB12/100 g in the BSG at lab scale. Finally, experiments were performed at 12 kg scale fermentation with a stabilized pH and resulted in 7.7 μg VitB12/100 g BSG, indicating the potential of in situ fortification of BSG with VitB12 by fermentation with P. freudenreichii. Overall, it was found that the VitB12-fortified BSG could serve as a relatively inexpensive plant-based source of VitB12-fortified protein for food and feed applications. This study shows that fermentation offers opportunities to fortify (protein-enriched) BSG or other BSG derivatives with VitB12.

Biosynthesis of propionic acid from whey permeate and corn steep liquor by Propionibacterium freudenreichii subsp ATCC 6207 and partial purification using ion exchange cryogels

Biosynthesis of propionic acid from whey permeate and corn steep liquor by Propionibacterium freudenreichii subsp ATCC 6207 and partial purification using ion exchange cryogels

Ethylene glycol is metabolized to ethanol and acetate and induces expression of bacterial microcompartments in Propionibacterium freudenreichii

Ethylene glycol (EG, 1,2-ethanediol) is a two-carbon dihydroxy alcohol that can be derived from fermentation of plant-derived xylose and arabinose and which can be formed during food fermentations. Here we show that Propionibacterium freudenreichii DSM 20271 is able to convert EG in anaerobic conditions to ethanol and acetate in almost equimolar amounts. The metabolism of EG led to a moderate increase of biomass, indicating its metabolism is energetically favourable. A proteomic analysis revealed EG induced expression of the pdu-cluster, which encodes a functional bacterial microcompartment (BMC) involved in the degradation of 1,2-propanediol, with the presence of BMCs confirmed using transmission electron microscopy. Cross-examination of the proteomes of 1,2-propanediol and EG grown cells revealed PDU BMC-expressing cells have elevated levels of DNA repair proteins and cysteine biosynthesis proteins. Cells grown in 1,2-propanediol and EG also showed enhanced resistance against acid and bile salt-induced stresses compared to lactate-grown cells. Our analysis of whole genome sequences of selected genomes of BMC-encoding microorganisms able to metabolize EG with acetaldehyde as intermediate indicate a potentially broad-distributed role of the pdu operon in metabolism of EG. Based on our results we conclude EG is metabolized to acetate and ethanol with acetaldehyde as intermediate within BMCs in P. freudenreichii.

Introducing Bacillus natto and Propionibacterium shermanii into soymilk fermentation: A promising strategy for quality improvement and bioactive peptide production during in vitro digestion

Herein, the texture properties, polyphenol contents, and in vitro protein digestion characteristics of soymilk single- or co-fermented by non-typical milk fermenter Bacillus natto (B. natto), Propionibacterium freudenreichii subsp. shermanii (P. shermanii), and traditional milk fermenter were evaluated. Co-fermenting procedure containing B. natto or P. shermanii could raise the amounts of gallic acid, caffeic acid, and GABA when compared to the unfermented soymilk. Co-fermented soymilk has higher in vitro protein digestibility and nutritional protein quality. Through peptidomic analysis, the co-work of P. shermanii and Lactobacillus plantarum (L. plantarum) may release the highest relative percentage of bioactive peptides, while the intervention of B. natto and Streptococcus thermophilus (S. thermophilus) resulted in more differentiated peptides. The multi-functional bioactive peptides were mainly released from glycine-rich protein, β-conglycinin alpha subunit 1, and ACB domain-containing protein. These findings indicated the potential usage of B. natto/S. thermophilus or P. shermanii/L. plantarum in bio-enhanced soymilk fermentation.

Oncom from Surplus Bread Enriched in Vitamin B12 via In Situ Production by Propionibacterium freudenreichii

Bread is a frequently wasted food product. Surplus or stale bread can be successfully processed by solid-state fermentation and used as the only fermentation substrate. Oncom, which originated in Indonesia, is made with moulds of the Neurospora genus. This experiment aimed to obtain oncome from stale bread enriched in vitamin B12. Co-fermentation with N. sitophila and Propionibacterium freudenreichii was carried out on two types of bread differing in chemical composition and initial pH value. Oncom obtained after 5 days of fermentation, depending on the substrate used and the fermentation variant (fungal, fungal-bacterial), contained from 35 to 40% dry mass, from 17.5 to about 23% protein, about 2 to max 5% fat, and from 65 to 74% carbohydrates by weight in dry mass. Vitamin B12 content depended largely on the bacterial strain, the colony-forming unit dose in the inoculum, and also the initial pH of the substrate. The oncom product obtained after co-fermentation with P. freudenreichii DSM 20271 contained a maximum of 1.3 µg/100 g, which corresponds to the vitamin B12 level in a chicken egg.

Short communication: a novel multispecies bacteria-based direct-fed microbial supports in vitro gut barrier integrity challenged with a pathogen or pro-inflammatory cytokines.

We conducted two experiments to evaluate the effects of a novel bacterial-based direct-fed microbial (DFM) on intestinal barrier integrity using the in vitro transepithelial electrical resistance (TEER) assay. In experiment 1, human-derived Caco-2 cells received or not (CON) a DFM containing Ligilactobacillus (formerly Lactobacillus) animalis 506, Propionibacterium freudenreichii 507, Bacillus paralicheniformis 809, and B. subtilis 597 (BDP; BOVAMINE DEFEND® Plus) at a rate of 1 × 108 CFU/transwell. Concurrently with treatment application (CON or BDP), a pathogenic challenge of Clostridium perfringens type A was added alone (PAT) or with BDP (PAT + BDP) at a rate of 2.8 × 107 CFU/transwell in a 2 × 2 factorial arrangement. In experiment 2, Caco-2 cells were also assigned in a 2 × 2 factorial design to CON or BDP and then, 2h post-treatment administration (CON and BDP), a mixture of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) was added alone (CYT) or with BDP (CYT + BDP) at a 10:1 ratio, respectively. In both experiments, TEER was measured for 18h. In experiment 1, a DFM × pathogen × hour interaction was observed for TEER (P < 0.0001). Adding the PAT alone initially tended to increase TEER vs. CON from 1.1 to 2.2h (P ≤ 0.09), increased TEER at 3.2h (P < 0.01), but reduced TEER from 5.4 to the end of the experimental period at 18.4h (P ≤ 0.01). On the other hand, adding DFM, with or without the pathogenic challenge, yielded greater TEER vs. CON-CON and CON-PAT for most of the experimental period (P ≤ 0.04). A similar interaction was detected and reported in experiment 2 (P < 0.0001). The CYT challenge reduced mean TEER compared with all other treatments from 3.2h to the remainder of the study (P ≤ 0.03). On the other hand, BDP-CYT was able to maintain the integrity of the epithelial cells when compared with CON-CON throughout the experimental period (P ≤ 0.03), the exception being at 3.2h (P = 0.20). Moreover, BDP-CON increased (P ≤ 0.04) TEER when compared with CON-CON from 3.2 to 18.4h, but also in comparison with BDP-CYT from 4.3 to 18.4h post-DFM and challenge administration into the cells. In summary, C. perfringens type A and a pro-inflammatory cytokine cocktail compromised the integrity of intestinal epithelial cell monolayers in vitro, whereas adding a multispecies bacteria-based DFM counteracted these damaging effects.

Co-culturing Propionibacterium freudenreichii and Bifidobacterium animalis subsp. lactis improves short-chain fatty acids and vitamin B12 contents in soy whey

The lack of vitamin B12 in unprocessed plant-based foods can lead to health problems in strict vegetarians and vegans. The main aim of this study was to investigate the potential synergy of co-culturing Bifidobacterium animalis subsp. lactis and Propionibacterium freudenreichii in improving production of vitamin B12 and short-chain fatty acids in soy whey. Different strategies including mono-, sequential and simultaneous cultures were adopted. Growth, short-chain fatty acids and vitamin B12 were assessed throughout the fermentation while free amino acids, volatiles, and isoflavones were determined on the final day. P. freudenreichii monoculture grew well in soy whey, whereas B. lactis monoculture entered the death phase by day 4. Principal component analysis demonstrates that metabolic changes in both sequential cultures did not show drastic differences to those of P. freudenreichii monoculture. However, simultaneous culturing significantly improved vitamin B12, acetic acid and propionic acid contents (1.3 times, 5 times, 2.5 times, compared to the next highest treatment [sequential cultures]) in fermented soy whey relative to other culturing modes. Hence, co-culturing of P. freudenreichii and B. lactis would provide an alternative method to improve vitamin B12, acetic acid and propionic acid contents in fermented foods.