Lactobacillus pentosus SMB718 as a probiotic starter producing allyl mercaptan in garlic and onion-enriched fermentation.

Fermenting garlic and onion provides the advantages of storage life extension, anti-oxidative and anti-diabetic activities, and their metabolite, allyl mercaptan, offers a strong aroma and various health benefits. Here, we report the probiotic properties of Lactobacillus pentosus SMB718 isolated from Korean traditional paste and its high allyl mercaptan productivity in garlic and onion fermentation. This strain was safe for use in food fermentation, as it was a non-biogenic amine producer and non-hemolytic. It showed high stability under simulated human gastrointestinal conditions and good adhesion ability to intestinal epithelial cells, including both Caco-2 and HT-29. This strain had antioxidant and anti-microbial activities. In addition, the heat-killed cells and lysate exerted anti-inflammatory effects on both LPS-stimulated RAW 264.7 cells and mouse macrophages by inhibition of pro-inflammatory cytokines and induction of anti-inflammatory cytokines. Furthermore, this strain possessed good fermentation properties in garlic and onion-enriched radish juice (GORJ); it grew well decreasing the pH and provided a rich aroma compound during fermentation. When L. pentosus SMB718 was used as a starter in garlic and onion fermentation, a significantly higher amount of allyl mercaptan (344 ppb) was produced compared with that produced by the type strain (82 ppb). In conclusion, L. pentosus SMB718 can be used as a beneficial probiotic starter for better flavor production in the fermentation of allium species plants.

Lactobacillus pentosus strain S-PT84 improves steatohepatitis by maintaining gut permeability.

Intestinal mucosal barrier dysfunction is closely related to the pathogenesis of nonalcoholic steatohepatitis (NASH). Gut immunity has been recently demonstrated to regulate gut barrier function. The Lactobacillus pentosus strain S-PT84 activates helper T cells and natural killer/natural killer T cells. In this study, we examined the effect of S-PT84 on NASH progression induced by high-cholesterol/high-fat diet (CL), focusing on the immune responses involved in gut barrier function. C57BL/6 mice were fed a normal chow or CL diet with or without 1 × 1010 S-PT84 for 22 weeks. S-PT84 administration improved hepatic steatosis by decreasing triglyceride and free fatty acid levels by 34% and 37%, respectively. Furthermore, S-PT84 inhibited the development of hepatic inflammation and fibrosis, suppressed F4/80+ macrophage/Kupffer cell infiltration, and reduced liver hydroxyproline content. Administration of S-PT84 alleviated hyperinsulinemia and enhanced hepatic insulin signalling. Compared with mice fed CL diet, mice fed CL+S-PT84 had 71% more CD11c-CD206+ M2 macrophages, resulting in a significantly decreased M1/M2 macrophage ratio in the liver. Moreover, S-PT84 inhibited the CL diet-mediated increase in intestinal permeability. Additionally, S-PT84 reduced the recruitment of interleukin-17-producing T cells and increased the levels of intestinal tight junction proteins, including zonula occludens-1, occludin, claudin-3, and claudin-7. In conclusion, our findings suggest that S-PT84 attenuates diet-induced insulin resistance and subsequent NASH development by maintaining gut permeability. Thus, S-PT84 represents a feasible approach to prevent the development of NASH.

Enhanced lactic acid bacteria viability with yeast coincubation under acidic conditions.

The enhancing effects of yeasts on the viability of lactic acid bacteria (LAB) under acidic conditions were investigated. Meyerozyma guilliermondii, coaggregative with both LAB strains under acidic conditions, significantly enhanced the viability of Lactobacillus pentosus and L. paracasei in pH 3.0 lactic acid (LA) buffer at 10°C (p < 0.05). Non-coaggregative yeasts (Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Cyberlindnera saturnus) also significantly enhanced the LAB viability (p < 0.05), and physical contact between LAB and yeasts was not essential for the viability-enhancing effect, indicating that the coaggregation had no relation to the enhancing mechanism. Although yeast metabolites and LA assimilation had no enhancing effect, hydrogen peroxide (H2O2) decreased after yeast coincubation, and H2O2 elimination improved L. pentosus viability. H2O2 elimination alone did not sufficiently improve L. paracasei viability, but the addition of antioxidants was effective. These results suggest that the antioxidant activity of yeast increased the LAB viability under acidic conditions.

Lactobacillus pentosus S-PT84 Prevents Low-Grade Chronic Inflammation-Associated Metabolic Disorders in a Lipopolysaccharide and High-Fat Diet C57/BL6J Mouse Model.

A long-term exposure to lipopolysaccharides results in the gut inflammation and its impaired barrier function, leading to the development of metabolic disorders. In this study, the role of dietary heat killed Lactobacillus pentosus S-PT84 on preventing endotoxemia to maintain metabolic homeostasis was studied. We demonstrated that the treatment of L. pentosus S-PT84 improved the gut integrity by maintaining tight-junction protein expression, in order to suppress the infiltration of endotoxin into plasma. The systemic inflammatory responses were inhibited via reducing the secretion of TNF-α and MCP-1. Furthermore, the blood lipid profile and glucose level as well as adiponectin in both plasma and white adipose tissues (WAT) were preserved by L. pentosus S-PT84 through upregulation of PPAR-γ and IRS-1 expression in WAT. The above findings suggest that the metabolic homeostasis in mice treated with HFD and LPS was sustained by L. pentosus S-PT84, leading to reducing the early risk for progression into metabolic disorders.

Enhanced viability of layer-by-layer encapsulated Lactobacillus pentosus using chitosan and sodium phytate.

Lactobacillus pentosus (LP) are widely used as probiotics in food products, dietary supplements, and nutraceuticals due to their health-promoting effects. To confer a functional effect, the probiotics need to survive during shelf-life and transit through the high acidic conditions of the stomach and bile salts in the small intestine. Herein, LP was firstly encapsulated in a layer-by-layer approach using chitosan (CS) and sodium phytate (SP). After digestion in simulated gastrointestinal fluid (SGF) for 120 min and 4% bile salts for 3 h, plain-LP exhibited a 7.40 and 6.09 colony forming units/ml (cfu/ml) reduction. Interestingly, two layer coated LP ((CS/SP)2-LP) exhibited less death, which reduced 4.34 and 2.33 log cfu/ml, respectively. Specially, (CS/SP)2-LP also showed a higher survival rate compared to plain-LP in heat treatment experiments, especially 65 °C. In conclusion, layer-by-layer encapsulation of LP has great potential for the protection and delivery of probiotics in food and nutraceutical products.

Exploring the influence of the surface proteins on probiotic effects performed by Lactobacillus pentosus HC-2 using transcriptome analysis in Litopenaeus vannamei midgut.

In order to understand the mediation function of surface proteins in probiotic effects executed by Lactobacillus pentosus HC-2 in midgut of Litopenaeus vannamei, the immune and digestion related enzymes and the transcriptome expression were analyzed after shrimp fed with normal HC-2 or with stripped surface proteins HC-2 by lithium chloride (LiCl) treatment. The results showed that the shrimp fed with normal HC-2 produced much higher immune and digestion related enzymes than the control group or LiCl-treated HC-2 group to defense the Vibrio parahaemolyticus E1 infection. We obtained total over 275,099 unigenes from L. vannamei midgut, 981 genes were significant differentially expressed in normal HC-2 group compared with control, 1314 genes were significant differentially expressed in LiCl-treated HC-2 group compared with control, and 1689 genes were significant differentially expressed in LiCl-treated HC-2 group compared with normal HC-2 group. The GO/KEGG enrichment analysis of the significantly different genes demonstrated that L. vannamei fed with normal HC-2 induced immune-related, signal transduction, ion homeostasis, cell-cell adhesion, response stress/stimulus, vascular endothelial growth factor and peritrophin genes up-regulation, which were important genes involved in improving the shrimp intestine immune response, nutrition and growth performance, and bacteria adhesion and colonization, but these genes were suppressed in the midgut of shrimp fed with deprived surface proteins bacteria. Taken together, these results indicated that the surface proteins were essential for HC-2 executing probiotic effects in midgut of shrimp. Our data contribute to improve the current understanding of host - Lactobacillus interaction and the probiotic mechanisms in shrimps.

Optimization of a single chamber microbial fuel cell using Lactobacillus pentosus: Influence of design and operating parameters.

Microbial fuel cells (MFCs) have been receiving an increased attention over the last years due to their potential to combat two global problems: waste pollution and energy demand. Additionally, when a wastewater is used, MFCs can perform its treatment while recovering energy, leading to the possibility of energy-producing wastewater treatment plants, offsetting their operational costs. However, to overcome their current limitations (lower power outputs and higher costs), a clear understanding of the effect of operation and design parameters on its overall performance is mandatory. Therefore, the goal of this work was to evaluate the effect of operating conditions - batch cycle and yeast extract concentration, and design parameters - anode electrode area, membrane thickness and active area, on the overall performance of a single chamber MFC. The MFC operated with a pure culture of Lactobacillus pentosus and a synthetic wastewater based on a real dairy industry effluent. The overall performance was evaluated through the power output and the COD removal rate. Additionally, the biofilm formed at the anode electrode was characterized in terms of biomass, proteins and polysaccharides content. For the conditions used in this work, a maximum power density of 5.04 ±â€¯0.39 mW/m2 was achieved with an anode electrode area of 61 cm2, a batch cycle of 48 h, 50 mg/L of yeast extract and a Nafion 212 membrane with an active area of 25 cm2. The different conditions tested had a clear effect on the MFC energy production and biofilm characteristics, but not on the ability of L. pentosus to treat the dairy wastewater. The COD removal rates were in the range between 42% and 58%, for all the conditions tested.

Effect of Spanish-style processing steps and inoculation with Lactobacillus pentosus starter culture on the volatile composition of cv. Manzanilla green olives.

The effects of the main steps of Spanish-style processing (alkaline treatment and fermentation) on the volatile composition of cv. Manzanilla green olives were studied. Both spontaneous and controlled fermentations were considered. In the latter case, a Lactobacillus pentosus strain from green olive fermentation brine was used as starter culture. The volatile profile was determined by headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS). Most of the volatile compounds detected in fresh olives decreased or were undetected after alkaline treatment, while several compounds (mostly acetic acid, 2-methylbutanoic acid, and ethanol) were formed as a result of this treatment. Over 50 new volatile components, mostly esters and phenols, appeared as a result of fermentation. The most outstanding finding was a considerable increase in 4-ethyl phenol (almost 100-fold increase) in inoculated olives compared to the uninoculated product. However, a sensory panel did not find significant differences in odor perception.

Multiple mechanisms applied by Lactobacillus pentosus AT6 to mute the lethal effects of Salmonella in a mouse model.

Probiotics are now prevalent world-wide, as functional food supplements with many benefits for humans and animals, such as protective effects against pathogenic infection. We showed that oral supplementation of Lactobacillus pentosus AT6 (AT6) decreased the mortality rate of mice with Salmonella infection. A series of experiments showed that the protective effects of AT6 on mice involved multiple mechanisms, including (1) the inhibition of Salmonella Typhimurium growth by AT6 or its cell-free culture supernatants (CFCSs); (2) the reduction of the bacterial loads of Salmonella Typhimurium in intestinal contents and internal organs, such as the liver and spleen; (3) the inhibition of adhesion and invasion of Salmonella Typhimurium into intestinal epithelial cells; and (4) the regulation of host immunities by modifying the production of a chain of cytokines. In conclusion, AT6 inhibited Salmonella infection via multiple mechanisms and therefore has great potential for the development of functional foods with anti-Salmonella activities.

In vitro investigation on probiotic, anti-Candida, and antibiofilm properties of Lactobacillus pentosus strain LAP1.

OBJECTIVE: To investigate the probiotic characteristics, anti-Candida activity, and antibiofilm attributes of Hentak derived Lactobacillus pentosus strain LAP1. DESIGN: The probiotic properties of strain LAP1 were depicted by adapting standard protocols. The anti-Candida and antibiofilm properties of isolate were determined using agar well diffusion assay and ELISA reader test, respectively. The time-kill assay was performed using viable colony count assay. Further, the co-aggregation property of strain LAP1 was determined based on standard methodology. RESULTS: Strain LAP1 exhibited not only tolerance to acidic pH but also showed resistivity (P ≤ 0.05) to simulated gastric juice exposure. Similarly, the strain was able to tolerate bile salt, showed hyperproteolytic activity, and also depicted susceptibility to most of the antibiotics tested. Auto-aggregation phenomenon (37.5-60%), hydrophobicity nature (42.85%), and survival potentiality of strain LAP1 under freeze-dried condition (9.0 ±â€¯0.01 log CFU/ml) made the isolate a promising probiotic candidate. Cell-free neutralized supernatant (CFNS) of strain LAP1 exhibited potent antifungal activities against C. albicans, C. tropicalis, and C. krusei with arbitrary unit of 150 ±â€¯4.34, 200 ±â€¯5.21, and 130 ±â€¯5.13 AU/ml, respectively and depicted remarkable reduction in the biofilm formation of respective Candida sp. in a concentration dependent manner. Moreover, time-kill assay data provided the growth inhibition of all Candida sp. in a time dependent manner. Additionally, strain LAP1 revealed significant co-aggregate percentage with C. albicans, C. tropicalis, and C. krusei. CONCLUSIONS: L. pentosus strain LAP1 exhibited a good probiotic characteristics, potent anti-Candida activity, and significant antibiofilm property that could be undoubtedly recommended for its vast applications not only in food industries but also as biotherapeutic agent against Candida infections in pharmaceutical industries.

The Behavior of Staphylococcus aureus Dual-Species Biofilms Treated with Bacteriophage phiIPLA-RODI Depends on the Accompanying Microorganism.

The use of bacteriophages as antimicrobials against pathogenic bacteria offers a promising alternative to traditional antibiotics and disinfectants. Significantly, phages may help to remove biofilms, which are notoriously resistant to commonly used eradication methods. However, the successful development of novel antibiofilm strategies must take into account that real-life biofilms usually consist of mixed-species populations. Within this context, this study aimed to explore the effectiveness of bacteriophage-based sanitation procedures for removing polymicrobial biofilms from food industry surfaces. We treated dual-species biofilms formed by the food pathogenic bacterium Staphylococcus aureus in combination with Lactobacillus plantarum, Enterococcus faecium, or Lactobacillus pentosus with the staphylococcal phage phiIPLA-RODI. Our results suggest that the impact of bacteriophage treatment on S. aureus mixed-species biofilms varies depending on the accompanying species and the infection conditions. For instance, short treatments (4 h) with a phage suspension under nutrient-limiting conditions reduced the number of S. aureus cells in 5-h biofilms by ∼1 log unit without releasing the nonsusceptible species. In contrast, longer infection periods (18 h) with no nutrient limitation increased the killing of S. aureus cells by the phage (decrease of up to 2.9 log units). However, in some cases, these conditions promoted the growth of the accompanying species. For example, the L. plantarum cell count in the treated sample was up to 2.3 log units higher than that in the untreated control. Furthermore, phage propagation inside dual-species biofilms also depended greatly on the accompanying species, with the highest rate detected in biofilms formed by S. aureus-L. pentosus Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) also showed changes in the three-dimensional structures of the mixed-species biofilms after phage treatment. Altogether, the results presented here highlight the need to study the impact of phage therapy on microbial communities that reflect a more realistic setting. IMPORTANCE: Biofilms represent a major source of contamination in industrial and hospital settings. Therefore, developing efficient strategies to combat bacterial biofilms is of the utmost importance from medical and economic perspectives. Bacteriophages have shown potential as novel antibiofilm agents, but further research is still required to fully understand the interactions between phages and biofilm-embedded bacteria. The results presented in this study contribute to achieving a better understanding of such interactions in a more realistic context, considering that most biofilms in the environment consist of mixed-species populations.