Effects of Lactobacillus reuteri MG5346 on Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL)-Induced Osteoclastogenesis and Ligature-Induced Experimental Periodontitis Rats.

Effects of culture supernatants of Lactobacillus reuteri MG5346 (CS-MG5346) on receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis were examined. CS-MG5346 treatment up to 400 µg/mL significantly reduced tartrate-resistant acid-phosphatase (TRAP) activity, the phenotype biomarker of osteoclast, without affecting cell viability. CS-MG5346 inhibited the expression of osteoclast specific transcriptional factors (c-fos and nuclear factor-activated T cells c1) and their target genes (TRAP, cathepsin, and matrix metallo-proteinase-9) in a dose-dependent manner (p<0.05). The administration of L. reuteri MG5346 (2×108 CFU/day) for 8 wks significantly improved furcation involvement, but no difference was observed in alveolar bone loss in ligature-induced experimental periodontitis rats. The elevated RANKL/ osteoprotegerin ratio, the biomarker of periodontitis, was significantly lowered in the gingival tissue by administration of L. reuteri MG5346 (p<0.05). L. reuteri MG5346 showed excellent stability in simulated stomach and intestinal fluids and did not have antibiotic resistance. Based on the results, L. reuteri MG5346 has the potential to be a promising probiotic strain for oral health.

Latilactobacillus sakei Wikim0066 Protects Skin through MMP Regulation on UVB-Irradiated In Vitro and In Vivo Model.

Ultraviolet (UV) B exposure induces wrinkle formation, collagen fiber breakdown, and transepidermal water loss (TEWL). UVB irradiation induces the expression of mitogen-activated protein kinase (MAPK), activator protein 1 (AP-1), and nuclear factor kappa B (NF-κB), which affect the expression of matrix metalloproteinases (MMP). We confirmed the effects of Latilactobacillus sakei wikim0066 (wikim0066) on UVB-irradiated Hs68 cells and HR-1 hairless mice cells. wikim0066 restored the production of type I procollagen by regulating the expression of MMP-1 and -3, MAPK, AP-1, and NF-κB in UVB-irradiated Hs68 cells and HR-1 mice. Oral administration of wikim0066 alleviates wrinkle formation, epidermal thickness, and TEWL in UVB-irradiated HR-1 hairless mice. These results indicated that wikim0066 has the potential to prevent UVB-induced wrinkle formation.

Efficacy and Safety of MED-01 Probiotics on Vaginal Health: A 12-Week, Multicenter, Randomized, Double-Blind, Placebo-Controlled Clinical Trial.

Bacterial vaginosis (BV) is the most common disease in women of childbearing age and is caused by the growth of abnormal microbiota in the vagina. Probiotic consumption can be an effective alternative treatment to preserve or improve vaginal health. In the present study, MED-01, a complex of five strains of probiotic candidates isolated from the vagina of Korean women, was used. This study was designed as a 12-week, randomized, multicenter, double-blind, placebo-controlled clinical trial to evaluate the efficacy and safety of MED-01 on vaginal health. A total of 101 reproductive-aged women with a Nugent score of 4-6 took MED-01 (5.0 × 109 CFU) or a placebo once a day, and 76 participants completed the procedure. MED-01 significantly reduced the Nugent score compared with the placebo. Quantitative PCR analysis confirmed that Lactobacillus plantarum was significantly increased in the vagina, whereas harmful bacteria such as Mobiluncus spp., Gardnerella vaginalis, and Atopobium vaginae were suppressed after 12 weeks of MED-01 ingestion. No adverse events to the test food supplements were observed in the participants. These results confirmed that MED-01 can be used as a probiotic for treating BV, as it improves the vaginal microbiota.

Protective Effect of Bifidobacterium animalis subs. lactis MG741 as Probiotics against UVB-Exposed Fibroblasts and Hairless Mice.

Skin photoaging, which causes wrinkles, increased epidermal thickness, and rough skin texture, is induced by ultraviolet B (UVB) exposure. These symptoms by skin photoaging have been reported to be involved in the reduction of collagen by the expression of matrix metalloproteinases (MMPs) and activator protein-1 (AP-1). This study investigated the protective effects of Bifidobacterium animalis subsp. lactis MG741 (Bi. lactis MG741) in Hs-68 fibroblasts and hairless mice (HR-1) following UVB exposure. We demonstrated that the Bi. lactis MG741 reduces wrinkles and skin thickness by downregulating MMP-1 and MMP-3, phosphorylation of extracellular signal-regulated kinase (ERK), and c-FOS in fibroblasts and HR-1. Additionally, in UVB-irradiated dorsal skin of HR-1, Bi. lactis MG741 inhibits the expression of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), an inflammation-related factor. Thus, Bi. lactis MG741 has the potential to prevent wrinkles and skin inflammation by modulating skin photoaging markers.

Lactic Acid Bacteria Improve the Photoprotective Effect via MAPK/AP-1/MMP Signaling Pathway on Skin Fibroblasts.

Ultraviolet B (UVB) exposure causes a breakdown of collagen, oxidative stress, and inflammation. UVB activates mitogen-activated protein kinase (MAPK), activator protein-1 (AP-1), and matrix metalloproteinases (MMPs). In this study, we evaluated 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) radical scavenging activity and the photoprotective effect of lactic acid bacteria LAB strains, including Lactobacillus, Bifidobacterium, and Streptococcus genera in UVB-exposed skin fibroblasts. Nine LAB strains displayed antioxidant activity by regulating superoxide dismutase in UVB-exposed skin fibroblasts. Four LAB strains (MG4684, MG5368, MG4511, and MG5140) recovered type I procollagen level by inhibiting MMPs, MAPK, and AP-1 protein expression. Additionally, these four strains reduced the expression of proinflammatory cytokines by inhibiting oxidative stress. Therefore, L. fermentum MG4684, MG5368, L. rhamnosus MG4511, and S. thermophilus MG5140 are potentially photoprotective.

Lacticaseibacillus rhamnosus MG4706 Suppresses Periodontitis in Osteoclasts, Inflammation-Inducing Cells, and Ligature-Induced Rats.

Periodontitis is a chronic inflammatory disease characterized by tooth loss due to inflammation and the loss of alveolar bone. Periodontitis is closely related to various systemic diseases and is emerging as a global health problem. In this study, we investigated the anti-inflammatory effect of lactic acid bacteria (LAB) in vitro on Porphyromonas gingivalis (P. gingivalis) LPS-activated RAW264.7 and human gingival fibroblasts-1 (HGF-1) cells and the anti-osteoclastogenic effect of LAB on RANKL-induced RAW264.7 cells. All LAB strains (Lacticaseibacillus rhamnosus MG4706, MG4709, and MG4711) inhibited nitric oxide (NO)/inducible nitric oxide synthase (iNOS) in P. gingivalis LPS-activated RAW264.7 cells and pro-inflammatory cytokines (IL-1ß and IL-6) and matrix metalloproteinase (MMP-8 and MMP-9) in HGF-1 cells. In addition, LAB treatment inhibited osteoclastogenesis by reducing tartrate-resistant acid phosphatase (TRAP) activity and cathepsin K (CtsK) through the downregulation of nuclear factor of activated T cells cytoplasmic 1 (NFATc1) and c-fos gene expression in RANKL-induced RAW264.7 cells. Administration of MG4706 alleviated alveolar bone loss indices and reduced the gene expression of IL-1ß, IL-6, MMP-8, MMP-9, and RANKL/OPG ratio in gingival tissue. In conclusion, L. rhamnosus MG4706 has the potential to alleviate periodontitis.

Evaluation of Safety and Probiotic Potential of Enterococcus faecalis MG5206 and Enterococcus faecium MG5232 Isolated from Kimchi, a Korean Fermented Cabbage.

The purpose of this study was to evaluate the genotypic and phenotypic toxicity of Enterococcus faecalis MG5206 and Enterococcus faecium MG5232 isolated from kimchi (fermented vegetable cabbage). In this study, the genotypic toxicity of the strains MG5206 and MG5232 was identified through whole-genome sequencing analysis, and phenotypic virulence, such as susceptibility to antibiotics, hemolytic activity, and gelatinase and hyaluronidase activities, was also evaluated. In addition, the in vivo toxicity of both strains was evaluated using an acute oral administration test in Sprague-Dawley rats. In all the tests, both the strains were determined to be safety by confirming that they did not show antibiotic resistance or virulence factors. In addition, these strains exhibited a low level of autoaggregation ability (37.2-66.3%) and hydrophobicity, as well as a high survival rate in gastrointestinal condition in vitro. Therefore, the safety and high gastrointestinal viability of E. faecalis MG5206 and E. faecium MG5232 suggests that both the strains could be utilized in food as potential probiotics in the future.

Lactobacilli Strain Mixture Alleviates Bacterial Vaginosis through Antibacterial and Antagonistic Activity in Gardnerella vaginalis-Infected C57BL/6 Mice.

The present study investigated the anti-bacterial vaginitis (BV) effects of a mixture of five lactobacilli strains (LM5), containing equal amounts of Ligilactobacillus salivarius MG242, Limosilactobacillus fermentum MG901, Lactiplantibacillus plantarum MG989, Lacticaseibacillus paracasei MG4272, and Lacticaseibacillus rhamnosus MG4288), in HeLa cells and Gardnerella vaginalis (GV)-infected BV mice. All strains produced lactic acid and hydrogen peroxide, and were resistant to nonoxynol-9. LM5 significantly inhibited GV growth by 80%, exhibited good adhesion to HeLa cells, and significantly inhibited GV adhesion to these cells. In GV-infected mice, LM5 administered orally at 5 × 109 CFU/mouse significantly inhibited GV proliferation in the vaginal tract and significantly reduced myeloperoxidase activity, pro-inflammatory cytokine (TNF-α, IL-1ß, and IL-6) levels, and nitric oxide levels in vaginal tissue lysates. Histopathological analysis of vaginal tissues revealed that LM5 markedly suppressed the exfoliation of vaginal epithelial cells. Overall, these results suggest that LM5 might alleviate BV by direct antibacterial and antagonistic activity in vaginal tissues of GV-infected mice.

In Vivo Confirmation of the Antimicrobial Effect of Probiotic Candidates against Gardnerella vaginalis.

Bacterial vaginosis (BV) is caused by a microbial imbalance of the vaginal ecosystem, causing genital discomfort and potentially even various complications in women. Moreover, research on the treatment or prevention of BV is increasing. In this study, we evaluated the antimicrobial and anti-inflammation effects of the lactic acid bacteria (LAB) Ligilactobacillus salivarius MG242, Limosilactobacillus fermentum MG901, and Lactiplantibacillus plantarum MG989 in a BV-induced mice model. The oral administration of the LAB significantly inhibited the growth of Gardnerella vaginalis up to 43% (p < 0.05). The LAB downregulated the expression of pro-inflammatory cytokines (IL-1ß and TNF-α) and myeloperoxidase (p < 0.05). Upon histological examination, the exfoliation of epithelial cells in the vaginal tissues was found to be reduced in the probiotic administration group compared to the infected group. In addition, the LAB tolerated the gastric and/or intestinal simulated conditions and proliferated, showing potential in promoting health based on hemolysis activity, antibiotic susceptibility, enzyme activity, and lactic acid production. Altogether, our results showed that the investigated LAB may be a good food ingredient candidate for ameliorating BV in women.

Inhibition of Nitric Oxide Production, Oxidative Stress Prevention, and Probiotic Activity of Lactic Acid Bacteria Isolated from the Human Vagina and Fermented Food.

In this study, lactic acid bacteria (LAB) with antioxidative and probiotic activities were isolated from the vaginas of Korean women and from fermented food. Among 34 isolated LAB strains, four strains (MG4221, MG4231, MG4261, and MG4270) exhibited inhibitory activity against nitric oxide production. The MG4221 and MG4270 strains were identified as Lactobacillus plantarum, and MG4231 and MG4261 were identified as Lactobacillus fermentum. These strains were able to tolerate pepsin and pancreatin, which is required for probiotic potential. The antioxidant effects of culture filtrates obtained from selected strains included 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging capacity. Most of the culture filtrates had effective DPPH scavenging activity.In conclusion, the selected strains have significant activities and are potentially applicable to the development of functional foods. These strains might also contribute to the prevention and control of several diseases associated with oxidative stress, when used as functional probiotics.

In vitro probiotic properties of vaginal Lactobacillus fermentum MG901 and Lactobacillus plantarum MG989 against Candida albicans.

Candida albicans is the most important Candida species causing vulvovaginal candidiasis (VVC). We investigated the potential of the probiotic strains Lactobacillus fermentum MG901 and L. plantarum MG989 towards control of C. albiacns. Cell viability tests following co-culturing with lactobacilli revealed that C. albicans cells lost metabolic activity and were eventually killed. Further studies revealed that MG901 and MG989 had high surface hydrophobicity that enhanced its adhesion ability to epithelial cell. The MG901 and MG989 showed coaggregation with E. coli and C. albicans to affect their adhesion and colonization. The adhesion of MG901 and MG989 to HT-29 cell and its inhibition of E. coli and C. albicans adherence to these cells were demonstrated. These incidences provided evidence of the possible colonization of MG901 and MG989 that would prevent binding and growth of E. coli and C. albicans onto intestinal epithelial cells. Following daily administration of 108 CFU of viable MG901 and MG989 orally, the animals' feces were examined for bacterial excretion. The potential probiotic MG901 and MG989 were found to persist for up to 6 days in the feces of mice. In conclusion, L. fermentum MG901 and L. plantarum MG989 have the potential to inhibit the yeast growth, which could possibly have played an important role in helping to clear VVC in vivo.

In Vitro Probiotic Properties of Lactobacillus salivarius MG242 Isolated from Human Vagina.

Vulvovaginal candidiasis (VVC) is a very common infection worldwide that is mainly caused by Candida albicans. In a previous study, we showed that Lactobacillus salivarius MG242 has anti-Gardnerella vaginalis activity. In this study, we investigated the potential of using L. salivarius MG242 for biocontrol of C. albicans. In line with the results from a spot overlay assay, MG242 inhibited the growth of C. albicans by 99.99 ± 0.01% in co-culture, suggesting that L. salivarius MG242 has the potential to be developed into a probiotic formula to treat or prevent VVC. Accelerated storage tests using dehydrated live cell powder at 50, 60, and 70 °C were performed, and the results showed that immobilization with 10% skim milk effectively increased the thermal resistance of entrapped microorganisms, resulting in sevenfold longer shelf-life than the control (in PBS). Lower storage temperatures also increased the shelf-life up to 8.31 months.

Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from oysters in Korea.

Vibrio parahaemolyticus is the most prevalent gastroenteritis-causing pathogen in Korea and in some other Asian countries. It is frequently found in oysters and other seafood. This study monitored changes in the prevalence of V. parahaemolyticus and environmental parameters in oyster aquaculture environments in Korea. From June to October 2014, we tested oysters (Crassostrea gigas) from shellfish-harvesting areas off the west coast of Korea. These 71 isolates were the sum of 16 (22.5%), 19 (26.8%), 23 (32.4%), and 13 (18.3%) isolates collected in July, August, September, and October, respectively. These 71 isolates had the following profiles of resistance against 16 antibiotics: all isolates were resistant to ampicillin and vancomycin, and 52.2, 50.7, and 50.7% of isolates exhibited resistance to cephalothin, rifampin, and streptomycin, respectively. PCR analysis for the presence of the species-specific toxR gene confirmed that 38 (53.5%) of the total 71 isolated strains were positive for V. parahaemolyticus. In PCR analysis for virulence of V. parahaemolyticus, of the 71 isolates tested in the present study, only 38 (53.5%) were positive for the trh virulence gene and 71 (100%) was negative for the tdh virulence gene.

Antibiotic resistance of Vibrio harveyi isolated from seawater in Korea.

Vibrio harveyi is an opportunistic human pathogen that may cause gastroenteritis, severe necrotizing soft-tissue infections, and primary septicemia, with a potentially high rate of lethality. In this study, we isolated and characterized V. harveyi from seawater collected from the West Sea in Korea, including sites located near shellfish farms. For the initial isolation of putative V. harveyi, isolates were incubated on thiosulfate citrate bile salt sucrose agar plates for 24h, followed by selection of greenish colonies. Gram-negative and oxidase-positive colonies were subsequently confirmed by biochemical assays and the API 20E kit test system. Species-specific 16S rRNA and hemolysin genes were used to design V. harveyi-specific PCR primers. From 840 seawater samples, a total of 2 strains of V. harveyi were isolated from shellfish farm seawater. The two isolates were subjected to profiling against 16 antibiotics and found to be resistant to cephalothin, vancomycin, ampicillin, cefepime, cefotetan, and streptomycin.