New strain of Pediococcus pentosaceus alleviates ethanol-induced liver injury by modulating the gut microbiota and short-chain fatty acid metabolism

BACKGROUND Intestinal dysbiosis has been shown to be associated with the pathogenesis of alcoholic liver disease(ALD), which includes changes in the microbiota composition and bacterial overgrowth, but an effective microbe-based therapy is lacking. Pediococcus pentosaceus(P. pentosaceus) CGMCC 7049 is a newly isolated strain of probiotic that has been shown to be resistant to ethanol and bile salts. However, further studies are needed to determine whether P. pentosaceus exerts a protective effect on ALD and to elucidate the potential mechanism.AIM To evaluate the protective effect of the probiotic P. pentosaceus on ethanol-induced liver injury in mice.METHODS A new ethanol-resistant strain of P. pentosaceus CGMCC 7049 was isolated from healthy adults in our laboratory. The chronic plus binge model of experimental ALD was established to evaluate the protective effects. Twenty-eight C57BL/6 mice were randomly divided into three groups: The control group received a pairfed control diet and oral gavage with sterile phosphate buffered saline, the EtOH group received a ten-day Lieber-DeCarli diet containing 5% ethanol and oral gavage with phosphate buffered saline, and the P. pentosaceus group received a 5% ethanol Lieber-DeCarli diet but was treated with P. pentosaceus. One dose of isocaloric maltose dextrin or ethanol was administered by oral gavage on day 11, and the mice were sacrificed nine hours later. Blood and tissue samples(liver and gut) were harvested to evaluate gut barrier function and liver injury-related parameters. Fresh cecal contents were collected, gas chromatography–mass spectrometry was used to measure short-chain fatty acid(SCFA) concentrations, and the microbiota composition was analyzed using 16S rRNA gene sequencing.RESULTS The P. pentosaceus treatment improved ethanol-induced liver injury, with lower alanine aminotransferase, aspartate transaminase and triglyceride levels and decreased neutrophil infiltration. These changes were accompanied by decreased levels of endotoxin and inflammatory cytokines, including interleukin-5, tumor necrosis factor-α, granulocyte colony-stimulating factor, keratinocyte-derived protein chemokine, macrophage inflammatory protein-1α and monocyte chemoattractant protein-1. Ethanol feeding resulted in intestinal dysbiosis and gut barrier disruption, increased relative abundance of potentially pathogenic Escherichia and Staphylococcus, and the depletion of SCFA-producing bacteria, such as Prevotella, Faecalibacterium, and Clostridium. In contrast, P. pentosaceus administration increased the microbial diversity, restored the relative abundance of Lactobacillus, Pediococcus, Prevotella, Clostridium and Akkermansia and increased propionic acid and butyric acid production by modifying SCFA-producing bacteria. Furthermore, the levels of the tight junction protein ZO-1, mucin proteins(mucin [MUC]-1, MUC-2 and MUC-4) and the antimicrobial peptide Reg3β were increased after probiotic supplementation.CONCLUSION Based on these results, the new strain of P. pentosaceus alleviated ethanol-induced liver injury by reversing gut microbiota dysbiosis, regulating intestinal SCFA metabolism, improving intestinal barrier function, and reducing circulating levels of endotoxin and proinflammatory cytokines and chemokines. Thus, this strain is a potential probiotic treatment for ALD.

Survivability of freeze-dried probiotic Pediococcus pentosaceus strains GS4,GS17 and Lactobacillus gasseri(ATCC 19992)during storage with commonly used pharmaceutical excipients within a period of 120 days

Objective: To examine the survivability and stability of probiotic strains in presence and absence of pharmaceutical excipients for a long period of time at(4 ± 1)℃.Methods: The survival rates of probiotic strains, Pediococcus pentosaceus GS4(MTCC12683)(NCBI HM044322), GS17(NCBI KJ608061) and Lactobacillus gasseri(ATCC 19992), were evaluated. Probiotic strains were lyophilized individually and in combination of excipients(sorbitol, ascorbic acid, fructose and skim milk). The preparation was monitored for 120 d storing at(4 ± 1)℃. During storage, all the preparations were evaluated for viability and stability of probiotic properties like lactic acid production, antimicrobial effect, water activity, and adherence to epithelial cells.Results: Sorbitol, ascorbic acid and skim milk favoured the viability of freeze-dried cells and sustained probiotic properties during storage. Without excipients(control group),strains showed percentage of survivability not more than 70% while strains with excipients survived for 73%–93% for a long period of time.Conclusions: Commonly used excipients can be considered as a vehicle for delivering active principle in probiotic formulation and for sustaining the viability and stability of probiotic strains for a period of 120 d.

Prospective use of bacteriocinogenic Pediococcus pentosaceus as direct-fed microbial having methane reducing potential

Direct-fed microbials（DFM）, generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly（P〈0.05） higher in live pedicoccal culture（LPC） and dead pedicoccal culture（DPC） both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower（P〈0.05） with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse（0.07 mL CH4 mg–1 dry matter digestibility） with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.

筛选与评估减缓肠道发炎功效之益生菌

本篇利用葡聚糖硫酸(Dextran Sulfate Sodium,DSS)诱发小鼠肠道发炎的模式来筛选与评估益生菌在减缓肠道发炎与损伤的效果。实验将8周龄的BALB/c小鼠分为六组(n = 10):正常对照组(无DSS诱导)、负对照组(DSS诱导)、片球菌GKA4组(DSS + GKA4)、片球菌GKP4组(DSS + GKP4)、乳酸菌GKR1组(DSS + GKR1)、以及乳酸菌GK4组(DSS + GK4)。连续14天以相当于成人一天摄取1克的剂量管喂小鼠益生菌,并于第7天到第14天的饮水中加入3.5% DSS,藉以诱发肠道发炎。实验结果显示给予3.5% DSS后,片球菌GKA4组及GKP4组与负对照组相比下,其对于体重减轻、结肠长度、疾病活动指数等指标均有明显改善。此外,亦可降低血清中促炎细胞激素IL-1β、IL-6及TNF-α含量,藉此减缓由DSS引发的肠道发炎反应。综观上述结果,片球菌P. acidilactici GKA4与P. pentosaceus GKP4具有应用于肠道保护的益生菌潜力。