l-glycine and l-glutamic acid protect Pediococcus pentosaceus R1 against oxidative damage induced by hydrogen peroxide.

The effects of l-glycine (Gly) and l-glutamic acid (Glu) on oxidative damage induced by hydrogen peroxide (H2O2) in Pediococcus pentosaceus R1 were investigated. Gly and Glu significantly reduce the production of intracellular reactive oxygen species and the levels of malondialdehyde and carbonylated proteins and concomitantly increase ATP levels in P. pentosaceus R1 under H2O2-induced stress (P < 0.05). Transmission electron microscopy and atomic force microscopy of bacteria under H2O2-induced stress revealed that Gly and Glu suppress bacterial membrane deformation and cell damage. Gly exhibited stronger ferrous ion-chelating ability, whereas Glu has higher radical scavenging activities and reducing power (P < 0.05). The abilities of Gly and Glu to inhibit lipid peroxidation are comparable. Gly and Glu significantly enhance the activities of superoxide dismutase and glutathione peroxidase, respectively, and increase the total antioxidant capacity of bacteria (P < 0.05). These findings indicate that Gly and Glu alleviate H2O2-induced oxidative stress via direct antioxidant effects and increase the activities of bacterial antioxidant enzyme.

Enhanced lactic acid production from P2O5-pretreated biomass by domesticated Pediococcus pentosaceus without detoxification.

Expensive cellulase and complex detoxification procedures increase the cost of biomass lactic acid fermentation. Therefore, it is of great significance to develop a robust method to ferment lactic acid using biomass by avoiding cellulase and detoxification. This study demonstrates the advantage of combining mechanocatalytic P2O5 pre-treatment and strain domestication. We show that an enzyme-free mechanocatalytic saccharification process by combining mix-milling of P2O5 with biomass and successive hydrolysis produces a fermentable hydrolysate with much less inhibitory compounds than the hydrolysates obtained by conventional methods; only 5-HMF, furfural and acetic acid were detected in the biomass hydrolysate, and no phenolic inhibitors were detected. Pretreatment of biomass with P2O5 not only avoided cellulase, but also obtained less toxic hydrolysate. Furthermore, the Pediococcus pentosaceus strain gained superior inhibitor tolerance through domestication. It could tolerate 17.1 g/L acetic acid, 12.5 g/L 5-HMF, 11.9 g/L guaiacol and 11.5 g/L furfural and showed activity in decomposing furfural and 5-HMF for self-detoxification, allowing efficient lactic acid fermentation from biomass hydrolysate without detoxification. The lactic acid concentration and conversion rate fermented by domesticated bacteria were increased by 113.5% and 22.4%, respectively. In addition, the domesticated bacteria could utilize glucose and xylose simultaneously to produce lactic acid selectively. The combination of P2O5 pre-treatment and strain domestication to ferment lactic acid is applied to several biomass feedstocks, including corn stalk, corn stalk residue and rice husk residue. Lactic acid concentrations of 29.8 g/L, 31.1 g/L, and 46.2 g/L were produced from the hydrolysates of corn stalk, corn stalk residue and rice husk residue, respectively.

Heterogeneity of macrolide-lincosamide-streptogramin phenotype & conjugal transfer of erm(B) in Pediococcus pentosaceus.

Background & objectives: Pediococcus pentosaceus has been reported to cause clinical infections while it is being promoted as probiotic in food formulations. Antibiotic resistance (AR) genes in this species are a matter of concern for treating clinical infections. The present study was aimed at understanding the phenotypic resistance of P. pentosaceus to macrolide-lincosamide-streptogramin B (MLSB) antibiotics and the transfer of AR to pathogens. Methods: P. pentosacues isolates (n=15) recovered from fermented foods were screened for phenotypic resistance to MLSBantibiotics using disc diffusion and microbroth dilution methods. Localization and transferability of the identified resistance genes, erm(B) and msr(C) were evaluated through Southern hybridization and in vitro conjugation methods. Results: Four different phenotypes; sensitive (S) (n=5), macrolide (M) (n=7), lincosamide (L) (n=2) and constitutive (cMLSB) (n=1) were observed among the 15 P. pentosaceus isolates. High-level resistance (>256 µg/ml) to MLSBwas observed with one cMLSBphenotypic isolate IB6-2A. Intermediate resistance (8-16 µg/ml) to macrolides and lincosamides was observed among M and L phenotype isolates, respectively. Cultures with S phenotype were susceptible to all other antibiotics but showed unusual minimum inhibitory concentration (MIC) values of 8-16 µg/ml for azithromycin. Southern hybridization studies revealed that resistance genes localized on the plasmids could be conjugally transferred to Enterococcus faecalis JH2-2. Interpretation & conclusions: The study provides insights into the emerging novel resistance patterns in P. pentosaceus and their ability to disseminate AR. Monitoring their resistance phenotypes before use of MLS antibiotics can help in successful treatment of Pediococcal infections in humans.

Agrimonia procera exerts antimicrobial effects, modulates the expression of defensins and cytokines in colonocytes and increases the immune response in lipopolysaccharide-challenged piglets.

BACKGROUND: Because antibiotic use in livestock is assumed to contribute to the emerging public health crisis of antibiotic resistance, alternatives are required. Phytogenic additives are extensively studied due to their antibiotic properties. Components of Agrimonia species have been reported as candidate antimicrobials that possess antioxidative and anti-inflammatory properties. We studied the impact of Agrimonia procera (AP) on the growth of selected strains of gut bacteria, the effect of AP on the mRNA abundance of genes involved in inflammation and bacterial defense in a colon carcinoma cell line, the effect of AP in piglets challenged with lipopolysaccharides, and the effect of AP on the growth performance of healthy piglets. RESULTS: The in vitro growth rate of different bacteria strains was negatively affected by AP, especially in Pediococcus pentosaceus and all tested E. coli strains. Stimulation of Caco-2 cells with TNFα resulted in elevated mRNA expression of CXCL1, IL-8 and GPX2. After pretreatment of cells with AP, stimulation of Caco-2 cells with TNFα still resulted in elevated mRNA expression of CXCL1 and IL-8 at all measured points in time. However, mRNA expression in AP-pretreated cells was lower after 6 h and 24 h. In addition, expression of DEFB1 and GPX2 was significantly elevated after TNFα stimulation. In vivo, application of lipopolysaccharides induced significantly increased animal body temperatures. Piglets pretreated with AP prior to lipopolysaccharide application showed a faster and larger increase in body temperature than controls. In addition, piglets pretreated with AP appeared to release more TNFα than controls. In healthy piglets, AP treatment had no impact on growth performance parameters. Fecal dry matter and total plasma antioxidant capacity tended to be higher in piglets treated with AP than in control piglets (P = 0.055 and P = 0.087, respectively). CONCLUSIONS: AP has antimicrobial effects in vitro and stimulated the expression of proinflammatory cytokines in Caco-2 cells. The additive had no effect on growth in healthy piglets but increased the immune response in LPS-treated animals. In addition, AP appeared to have antioxidative effects in vivo. Therefore, AP merits testing as a future alternative to antibiotics in animal husbandry.

Prebiotic effects of bovine lactoferrin on specific probiotic bacteria.

Bovine lactoferrin (bLf) is a natural iron-binding protein and it has been suggested to be a prebiotic agent, but this finding remains inconclusive. This study explores the prebiotic potential of bLf in 14 probiotics. Initially, bLf (1-32 mg/mL) treatment showed occasional and slight prebiotic activity in several probiotics only during the late experimental period (48, 78 h) at 37 °C. We subsequently supposed that bLf exerts stronger prebiotic effects when probiotic growth has been temperately retarded. Therefore, we incubated the probiotics at different temperatures, namely 37 °C, 28 °C, room temperature (approximately 22-24 °C), and 22 °C, to retard or inhibit their growth. As expected, bLf showed more favorable prebiotic activity in several probiotics when their growth was partially retarded at room temperature. Furthermore, at 22 °C, the growth of Bifidobacterium breve, Lactobacillus coryniformis, L. delbrueckii, L. acidophilus, B. angulatum, B. catenulatum, and L. paraplantarum were completely blocked. Notably, these probiotics started regrowing in the presence of bLf (1-32 mg/mL) in a significant and dose-dependent manner. Accordingly, bLf significantly increased the growth of Pediococcus pentosaceus, L. rhamnosus, and L. paracasei (BCRC 17483; a locally isolated strain) when their growth was retarded by incubation at 22 °C. In conclusion, bLf showed inconsistent prebiotic activity in the 14 probiotics at 37 °C, but revealed strong prebiotic activity in 10 probiotic strains at 22 °C. Therefore, this study enables determining additional roles of Lf in probiotic strains, which can facilitate developing novel combinational approaches by simultaneously using Lf and specific probiotics.

Unraveling the probiotic efficiency of bacterium Pediococcus pentosaceus OBK05 isolated from buttermilk: An in vitro study for cholesterol assimilation potential and antibiotic resistance status.

The present study describes the probiotic potential and functional properties of the lactic acid bacterium Pediococcus pentosaceus OBK05 isolated from buttermilk. The isolate OBK05 was assessed for its probiotic properties. The isolate showed notable tolerance to pH 2.0 and 3.0 (8.44, 8.35 log CFU/mL), oxbile of 0.5% at 2 and 4 h of incubation (6.97, 6.35 log CFU/mL) and higher aggregation (auto-aggregation, adhesion to hydrocarbons) than the referral strain, Lactobacillus acidophilus MTCC 10307. The adhesion efficiency to HT-29 cells was found to be maximum, corresponding to 93.5% and 97% at 1 and 2 h incubation, respectively. In addition, the isolate OBK05 showed antagonistic solid activity against bacterial pathogens like Pseudomonas aeruginosa MTCC 424 and Bacillus subtilis MTCC 1133. The phenotypic antibiotic resistance of the isolate was examined before and after curing plasmids. Among the known five structural genes responsible for different antibiotic resistance, four genes indicating antibiotic resistance to kanamycin-Aph (3´´)-III, streptomycin-strA, vancomycin-vanA and ciprofloxacin-gyrA were detected by PCR amplification of genomic DNA. Further, the horizontal gene transfer from OBK05 isolate to pathogens was not found for these antibiotic resistance markers when filter and food mating were carried out as no transconjugants developed on media plates containing respective antibiotics. This indicates that the intrinsic resistance is harbored on chromosomal genes, and hence it is nontransferable to other microbes. In addition, strain OBK05 exhibited good DPPH scavenging activity of 56 to 77% and liberated free amino acid from conjugated bile acid. The strain OBK05 demonstrated a strong ability to reduce cholesterol at 12 h (17%), 24 h (27%) and 48 h (67%) of incubation.

Potential Control of Listeria monocytogenes by Bacteriocinogenic Enterococcus hirae ST57ACC and Pediococcus pentosaceus ST65ACC Strains Isolated From Artisanal Cheese.

Bacteriocinogenic Enterococcus hirae ST57ACC and Pediococcus pentosaceus ST65ACC strains, previously isolated from artisanal cheese, were evaluated for their safety with the aim to determine whether they could be used as beneficial strains, especially in the control of Listeria monocytogenes. Both isolates survived simulated gastrointestinal conditions and showed high levels of auto- and co-aggregation with L. monocytogenes, although the hydrophobicity of cells varied. Using the agar-spot test with 33 commercial drugs from different groups, only anti-inflammatory drugs and drugs containing loratadine and propranolol hydrochloride were able to affect the growth of the tested strains. Both strains were resistant to 3 out of 11 antibiotics tested by the disc diffusion method, and low frequencies of antibiotic resistance-encoding genes were observed by PCR analysis. Tested strains neither presented biogenic amine-related genes nor produced these substances. Aside from some antibiotic resistance characteristics, the tested strains were considered safe as they lack other virulence-related genes. E. hirae ST57ACC and P. pentosaceus ST65ACC both presented beneficial properties, particularly their ability to survive gastrointestinal conditions and to aggregate with L. monocytogenes, which can facilitate the elimination of this pathogen. Further studies should be conducted to better understand these interactions.

Infective endocarditis caused by Lactococcus lactis subsp. lactis and Pediococcus pentosaceus: A case report and literature review.

RATIONALE: Lactococcus lactis and Pediococcus pentosaceus are rare pathogens which rarely caused infections in humans. Several cases with L. lactis endocarditis have been reported in the literature, among them few were caused by L. lactis subsp. Lactis. Opportunistic P. pentosaceus infections were rarely reported. PATIENT CONCERNS: A 66-year-old man presented to our hospital due to persistent fever for 15 days. A physical checking revealed a grade II holosystolic murmur at the heart apex. A chest computed tomography (CT) scan suggested bronchitis. L. lactis subsp. lactis was identified in blood cultures. Transthoracic and transesophageal echocardiography revealed the presence of a large hyperechogenic mass in the left atrium, and a large floating vegetation on the mitral valve with a severe mitral regurgitation. DIAGNOSIS: Infectious endocarditis caused by L. lactis subsp. Lactis was diagnosed. INTERVENTIONS: Levofloxacin (0.5 g/day) was used for 20 days; however, L. lactis subsp. lactis remained to be isolated from blood culture. Therefore, vancomycin (2 g/day) was used to replace levofloxacin. Six days after the treatment with vancomycin, the blood culture revealed no L. lactis subspecies lactis, but yielded a growth of gram-positive and non-spore forming cocci; and P. pentosaceus was identified. Antimicrobial susceptibility test revealed P. pentosaceus was sensitive to penicillin and levofloxacin. Vancomycin was discontinued, and levofloxacin (0.5 g/day) was restarted and treated for another 7 days. The patient recovered with negative blood culture results, and discharged from the hospital. OUTCOMES: The patient recovered with negative blood culture results, and discharged from the hospital. LESSONS: Our patient had a long-period of antibiotic treatment with strategy alterations. Standard interpretation criteria of Clinical and Laboratory Standards Institute (CLSI) for L. lactis should be established, and molecular tests will increase the identification rate of L. lactis infections.