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Lactobacillus reuteri AN417 cell-free culture supernatant as a novel antibacterial agent targeting oral pathogenic bacteria.

ABSTRACT: Lactobacillus reuteri AN417 is a newly characterized probiotic strain. The activity of AN417 against oral pathogenic bacteria is unknown. We investigated the antibacterial activity of cell-free L. reuteri AN417 culture supernatant (LRS) against three oral pathogens: Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans. P. gingivalis and F. nucleatum have been implicated in periodontal disease, whereas S. mutans causes dental caries. Exposing these oral pathogenic bacteria to LRS significantly reduced their growth rates, intracellular ATP levels, cell viability, and time-to-kill. The minimal inhibitory volume of LRS was 10% (v/v) against P. gingivalis, 20% (v/v) for F. nucleatum, and 30% (v/v) for S. mutans. LRS significantly reduced the integrity of biofilms and significantly suppressed the expression of various genes involved in P. gingivalis biofilm formation. The L. reuteri AN417 genome lacked genes encoding reuterin, reuteran, and reutericyclin, which are major antibacterial compounds produced in L. reuteri strains. LRS treated with lipase and ?-amylase displayed decreased antibacterial activity against oral pathogens. These data suggest that the antibacterial substances in LRS are carbohydrates and/or fatty acid metabolites. Our results demonstrate that LRS has antimicrobial activity against dental pathogenic bacteria, highlighting its potential utility for the prevention and treatment of P. gingivalis periodontal disease.

SUBMITTER: Yang KM

PROVIDER: S-EPMC7810884 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Lactobacillus reuteri AN417 cell-free culture supernatant as a novel antibacterial agent targeting oral pathogenic bacteria.

Yang Kyung Mi KM Kim Ji-Sun JS Kim Hye-Sung HS Kim Young-Youn YY Oh Jeong-Kyu JK Jung Hye-Won HW Park Doo-Sang DS Bae Kwang-Hak KH

Scientific reports 20210115 1

Lactobacillus reuteri AN417 is a newly characterized probiotic strain. The activity of AN417 against oral pathogenic bacteria is unknown. We investigated the antibacterial activity of cell-free L. reuteri AN417 culture supernatant (LRS) against three oral pathogens: Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans. P. gingivalis and F. nucleatum have been implicated in periodontal disease, whereas S. mutans causes dental caries. Exposing these oral pathogenic bacteria ...[more]

PMID: 33452304

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Project description:BackgroundGut microbiota modulation has been demonstrated to be effective in protecting patients against detrimental effects of anti-cancer therapies, as well as to improve the efficacy of certain anti-cancer treatments. Among the most characterized probiotics, Lactobacillus rhamnosus GG (LGG) is currently utilized in clinics to alleviate diarrhea, mucositis or intestinal damage which might be associated with several triggers, including Clostridium difficile infections, inflammatory gut diseases, antibiotic consumption, chemotherapy or radiation therapy. Here, we investigate whether LGG cell-free supernatant (LGG-SN) might exert anti-proliferative activity toward colon cancer and metastatic melanoma cells. Moreover, we assess the potential adjuvant effect of LGG-SN in combination with anti-cancer drugs.MethodsLGG-SN alone or in combination with either 5-Fuorouracil and Irinotecan was used to treat human colon and human melanoma cancer cell lines. Dimethylimidazol-diphenyl tetrazolium bromide assay was employed to detect cellular viability. Trypan blue staining, anti-cleaved caspase-3 and anti-total versus anti-cleaved PARP western blots, and annexin V/propidium iodide flow cytometry analyses were used to assess cell death. Flow cytometry measurement of cellular DNA content (with propidium iodide staining) together with qPCR analysis of cyclins expression were used to assess cell cycle.ResultsWe demonstrate that LGG-SN is able to selectively reduce the viability of cancer cells in a concentration-dependent way. While LGG-SN does not exert any anti-proliferative activity on control fibroblasts. In cancer cells, the reduction in viability is not associated with apoptosis induction, but with a mitotic arrest in the G2/M phase of cell cycle. Additionally, LGG-SN sensitizes cancer cells to both 5-Fluorouracil and Irinotecan, thereby showing a positive synergistic action.ConclusionOverall, our results suggest that LGG-SN may contain one or more bioactive molecules with anti-cancer activity which sensitize cancer cells to chemotherapeutic drugs. Thus, LGG could be proposed as an ideal candidate for ground-breaking integrated approaches to be employed in oncology, to reduce chemotherapy-related side effects and overcome resistance or relapse issues, thus ameliorating the therapeutic response in cancer patients.

Project description:Purpose:In this study, silver nanoparticles (AgNPs) were biosynthesized using culture supernatant of strain Shewanella sp. ARY1, characterized and their antibacterial activity was investigated against Gram-negative bacteria Escherichia coli and Klebsiella pneumoniae. Methods:The strain Shewanella sp. ARY1 was isolated from river Yamuna, Delhi and used for biosynthesis of AgNPs via extracellular approach. Biosynthesized AgNPs were characterized by UV-Visible (UV-Vis) spectrophotometer, fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), energy dispersive X-ray (EDX), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Antibacterial activity of AgNPs was determined by well diffusion, broth microdilution and streaking plate assay to determine the zone of inhibition (ZOI), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), respectively. The effect of AgNPs on treated bacteria was investigated by electron microscopy analysis. Further, the biocompatibility of AgNPs was tested against mice erythrocytes (RBC) by hemolytic assay. Results:The UV-Vis spectral analysis revealed absorption maxima at 450 nm which confirmed the formation of AgNPs. The FTIR analysis suggested the involvement of various supernatant biomolecules, as reducing and capping agents in the synthesis of AgNPs. The XRD and EDX analysis confirmed the crystalline and metallic nature of AgNPs, respectively. The TEM and SEM analysis showed nanoparticles were spherical with an average size of 38 nm. The biosynthesized AgNPs inhibited the growth and formed a clear zone of inhibition (ZOI) against tested Gram-negative strains. The MIC and MBC were determined as 8-16 µg/mL and 32 µg/mL, respectively. Further, electron microscopy analysis of treated cells showed that AgNPs can damage the outer membrane, release of cytoplasmic contents, and alter the normal morphology of Gram-negative bacteria, leading to cell death. The hemolytic assay indicated that the biosynthesized AgNPs were biocompatible at low dose concentrations. Conclusion:This study demonstrates an eco-friendly process for extracellular synthesis of AgNPs using Shewanella sp. ARY1 and these AgNPs exhibited excellent antibacterial activity, which may be used to combat Gram-negative pathogens.

Dataset Information

Lactobacillus reuteri AN417 cell-free culture supernatant as a novel antibacterial agent targeting oral pathogenic bacteria.

Publications

Lactobacillus reuteri AN417 cell-free culture supernatant as a novel antibacterial agent targeting oral pathogenic bacteria.

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