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The effect of Lactobacillus reuteri cell free supernatant on growth and biofilm formation of Paenibacillus larvae.

ABSTRACT: Background:Paenibacillus larvae is the etiological agent of American foulbrood (AFB) disease, the most lethal disease in honeybee (Apis mellifera) larvae. Aims:The aim of the present work was to study the antimicrobial effect of cell free supernatant (CFS) of probiotics on an Iranian isolate's biofilm formation. Methods:A local strain was identified by 16S rRNA sequencing. The antibacterial effect of some probiotics was evaluated through drop plate method, minimum inhibitory concentrations (MIC), minimum bactericidal concentrations (MBC) and time-kill assay. The biofilm formation ability of P. larvae and the inhibition of biofilm formation by CFS were studied by microplate and scanning electron microscopy (SEM). The nature of the secondary metabolites in CFS was examined by microscale optical density assay (MODA). Results:Alignment of the results of P. larvae KB10 (GenBank accession number MH000685.1) 16S rRNA with the database revealed more than 97% identity with P. larvae . The most antibacterial effect was observed in the CFS of Lactobacillus reuteri ATCC23272 with 12.75 ± 3.2 mm for zone of inhibition (ZOI) at 1000 µL/ml for MIC and MBC. Time-kill assay revealed that CFS eliminated 1.5 × 108 CFU/ml P. larvae KB10 at 2 h of exposure. Microtitre plate and SEM results revealed that CFS (at sub-MIC concentration) was able to inhibit biofilm formation by P. larvae. The results of MODA assay showed that antimicrobial activity were related to the production of organic acids. Conclusion:Cell free supernatant from L. reuteri ATCC 23272 had inhibitory effects on P. larvae KB10 growth and biofilm production due to its acidic nature. The obtained results can be used for antibiotic substitution in AFB control and treatment.

SUBMITTER: Betesho Babrud R

PROVIDER: S-EPMC6811706 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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The effect of <i>Lactobacillus reuteri</i> cell free supernatant on growth and biofilm formation of <i>Paenibacillus larvae</i>.

Betesho Babrud R R Kasra Kermanshahi R R Motamedi Sede F F Moosavinejad S Z SZ

Iranian journal of veterinary research 20190101 3

<h4>Background</h4><i>Paenibacillus larvae</i> is the etiological agent of American foulbrood (AFB) disease, the most lethal disease in honeybee (<i>Apis mellifera</i>) larvae.<h4>Aims</h4>The aim of the present work was to study the antimicrobial effect of cell free supernatant (CFS) of probiotics on an Iranian isolate's biofilm formation.<h4>Methods</h4>A local strain was identified by <i>16S rRNA</i> sequencing. The antibacterial effect of some probiotics was evaluated through drop plate meth ...[more]

PMID: 31656524

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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:Non-typhoidal Salmonella (NTS) can cause infection in poultry, livestock, and humans. Although the use of antimicrobials as feed additives is prohibited, the previous indiscriminate use and poor regulatory oversight in some parts of the world have resulted in increased bacterial resistance to antimicrobials, including cephalosporins and fluoroquinolones, which are among the limited treatment options available against NTS. This study aimed to isolate potential probiotic lactic acid bacteria (LAB) strains from the poultry gut to inhibit fluoroquinolone and cephalosporin resistant MDR Salmonella Typhimurium and S. Enteritidis. The safety profile of the LAB isolates was evaluated for the hemolytic activity, DNase activity, and antibiotic resistance. Based on the safety results, three possible probiotic LAB candidates for in vitro Salmonella control were chosen. Candidate LAB isolates were identified by 16S rDNA sequencing as Lactobacillus reuteri PFS4, Enterococcus faecium PFS13, and Enterococcus faecium PFS14. These strains demonstrated a good tolerance to gastrointestinal-related stresses, including gastric acid, bile, lysozyme, and phenol. In addition, the isolates that were able to auto aggregate had the ability to co-aggregate with MDR S. Typhimurium and S. Enteritidis. Furthermore, LAB strains competitively reduced the adhesion of pathogens to porcine mucin Type III in co-culture studies. The probiotic combination of the selected LAB isolates inhibited the biofilm formation of S. Typhimurium FML15 and S. Enteritidis FML18 by 90% and 92%, respectively. In addition, the cell-free supernatant (CFS) of the LAB culture significantly reduced the growth of Salmonella in vitro. Thus, L. reuteri PFS4, E. faecium PFS13, and E. faecium PFS 14 are potential probiotics that could be used to control MDR S. Typhimurium and S. Enteritidis in poultry. Future investigations are required to elucidate the in vivo potential of these probiotic candidates as Salmonella control agents in poultry and animal feed.

Dataset Information

The effect of Lactobacillus reuteri cell free supernatant on growth and biofilm formation of Paenibacillus larvae.

Publications

The effect of <i>Lactobacillus reuteri</i> cell free supernatant on growth and biofilm formation of <i>Paenibacillus larvae</i>.

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