Gut microbiota, body weight and histopathological examinations in experimental infection by methicillin-resistant Staphylococcus aureus: antibiotic versus bacteriocin.

Bacteriocins have been steadily reported as potential agents that may contribute, in different ways, to overcome antimicrobial drug resistance. Here, holoxenic NMRI-F mice microbiota, their body weight recovery and histopathological alterations of organs like colon, spleen and liver were examined in mice intraperitoneally infected with 108 cfu of a clinical methicillin-resistant Staphylococcus aureus (MRSA-1), and treated with enterocin DD14 alone (165 mg/kg), erythromycin alone (100 mg/kg) or their combination. Animals that received both antimicrobials presented a better body weight recovery than other groups. Less pronounced histopathological alterations were observed in mice MRSA-infected and treated with bacteriocin than in those MRSA-infected but untreated or MRSA-infected and treated with erythromycin. Noteworthy, these alterations were absent when mice were treated with MRSA-infected and treated with both antibacterial agents. Furthermore, the genus richness was significantly lower in mice infected and treated with erythromycin, compared to mice infected and treated with both antimicrobials. The beta-diversity analysis showed that non-infected mice and those infected and treated with both antimicrobials, stand apart from the other groups as supported in a NMDS model. This in vivo study shows the relevance of bacteriocin, or bacteriocin-antibiotic formulation in protecting colonic, liver and spleen soft tissues and controlling the mouse gut microbiota, following MRSA infection.

Purification and characterization of the bacteriocin produced by Lactobacillus sakei MBSa1 isolated from Brazilian salami.

AIMS: The study aimed at determining the biochemical characteristics of the bacteriocin produced by Lactobacillus sakei MBSa1, isolated from salami, correlating the results with the genetic features of the producer strain. METHODS AND RESULTS: Identification of strain MBSa1 was performed by 16S rDNA sequencing. The bacteriocin was tested for spectrum of activity, heat and pH stability, mechanism of action, molecular mass and amino acid sequence when purified by cation-exchange and reversed-phase HPLC. Genomic DNA was tested for bacteriocin genes commonly present in Lact. sakei. Bacteriocin MBSa1 was heat-stable, unaffected by pH 2·0 to 6·0 and active against all tested Listeria monocytogenes strains. Maximal production of bacteriocin MBSa1 (1600 AU ml(-1)) in MRS broth occurred after 20 h at 25°C. The molecular mass of produced bacteriocin was 4303·3 Da, and the molecule contained the SIIGGMISGWAASGLAG sequence, also present in sakacin A. The strain contained the sakacin A and curvacin A genes but was negative for other tested sakacin genes (sakacins T-α, T-ß, X, P, G and Q). CONCLUSIONS: In the studied conditions, Lact. sakei MBSa1 produced sakacin A, a class II bacteriocin, with anti-Listeria activity. SIGNIFICANCE AND IMPACT OF THE STUDY: The study covers the purification and characterization of the bacteriocin produced by a lactic acid bacteria isolated from salami (Lact. sakei MBSa1), linking genetic and expression information. Its heat-resistance, pH stability in acid conditions (pH 2·0-6·0) and activity against L. monocytogenes food isolates bring up a potential technological application to improve food safety.

Antifungal properties of durancins isolated from Enterococcus durans A5-11 and of its synthetic fragments.

The aim of this work was to study the antifungal properties of durancins isolated from Enterococcus durans A5-11 and of their chemically synthesized fragments. Enterococcus durans A5-11 is a lactic acid bacteria strain isolated from traditional Mongolian airag cheese. This strain inhibits the growth of several fungi including Fusarium culmorum, Penicillium roqueforti and Debaryomyces hansenii. It produces two bacteriocins: durancin A5-11a and durancin A5-11b, which have similar antimicrobial properties. The whole durancins A5-11a and A5-11b, as well as their N- and C-terminal fragments were synthesized, and their antifungal properties were studied. C-terminal fragments of both durancins showed stronger antifungal activities than other tested peptides. Treatment of D. hansenii LMSA2.11.003 strain with 2 mmol l(-1) of the synthetic peptides led to the loss of the membrane integrity and to several changes in the ultra-structure of the yeast cells. Chemically synthesized durancins and their synthetic fragments showed different antimicrobial properties from each other. N-terminal peptides show activities against both bacterial and fungal strains tested. C-terminal peptides have specific activities against tested fungal strain and do not show antibacterial activity. However, the C-terminal fragment enhances the activity of the N-terminal fragment in the whole bacteriocins against bacteria.

Partial purification and characterization of the mode of action of enterocin S37: a bacteriocin produced by Enterococcus faecalis S37 isolated from poultry feces.

The aim of this research was to purify and characterize the mode of action of enterocin S37, a bacteriocin produced by Enterococcus faecalis S37, a strain recently isolated from the chicken feces. Enterocin S37 has a molecular weight comprised between 4 and 5 kDa. It remained active after 1 h at 80(o)C and at pH values ranging from 4.0 to 9.0. Furthermore, cell-free supernatant of Enterococcus faecalis S37 and purified enterocin S37 were active against Gram-positive bacteria including Listeria monocytogenes EGDe, L. innocua F, Enterococcus faecalis JH2-2, and Lactobacillus brevis F145. The purification of enterocin S37 was performed by ammonium sulfate precipitation followed up by hydrophobic-interaction chromatography procedures. Treatment of enterocin S37 with proteinase K, alpha-chymotrypsin, and papain confirmed its proteinaceous nature, while its treatment with lysozyme and lipase resulted in no alteration of activity. Enterocin S37 is hydrophobic, anti-Listeria and likely acting by depletion of intracellular K(+) ions upon action on K(ATP) channels. This study contributed to gain more insights into the mode of action of enterocins.

Divercin V41 from gene characterization to food applications: 1998-2008, a decade of solved and unsolved questions.

The emergence of an increasing number of antibiotic resistant human clinical bacteria has been a great cause of concern for the last decades. As an example, Staphylococcus aureus isolates in the hospital environment are becoming more and more resistant to antibiotics including vancomycin which is considered as a last line of defence in treatment of Staphylococcus aureus-resistant methicillin. On the other hand, food safety is threatened by development of pathogenic bacteria including Listeria monocytogenes, Campylobacter jejuni, Salmonella enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. The use of antimicrobial peptides such as glycopeptides, semi-synthetic peptides, bacteriocins including lantibiotics offers a hope to face these clinical and food microbiology concerns. Clinical approval of new chemotherapeutic agents requires a long period of time. Research on bacteriocins has demonstrated potential use to fight against undesired foodborne pathogens but the use industrial use of bacteriocins is limited. To date only lantibiotic nisin and in class IIa bacteriocin Pediocin PA-1 are legally used as food preservative in many countries. The present minireview is focused on divercin V41 (DvnV41), a class IIa bacteriocin naturally produced by Carnobacterium divergens V41. The last decade has been the witness of intensive investigations carried out on this cationic peptide tempting to answer multiple questions covering basic and applied aspects. DvnV41 has shown a wide spectrum of activity either alone or in combination with nisin and/or polymixins (synergistic effect). This outcome indicates that Cb. divergens V41 could potentially be used for safe and efficient prevention of L. monocytogenes growth in cold smoked salmon.

Identification of lactic acid bacteria from poultry feces: evidence on anti-campylobacter and anti-listeria activities.

Forty-five strains of lactic acid bacteria were isolated from poultry feces sampled from an industrial farm located in the Nantes area (France), and most of them belong to the Streptococcus or Lactobacillus genus according to the results obtained by Galerie API and molecular methods. The most representative lactic acid bacteria strains were screened for antibacterial compound production against 2 indicator organisms (Listeria innocua F and Campylobacter jejuni 11168) by means of the agar diffusion test. Strain S37, identified as Enterococcus faecalis, exhibited a clear anti-listerial activity and a slight anti-Campylobacter activity, whereas strain S42, identified as Lactobacillus reuteri, exhibited only anti-Campylobacter activity. Regarding the results of proteolytic, heat, and neutralizing treatments of supernatants from the aforementioned strains, we can conclude that antagonism observed is attributed to antimicrobial peptide or bacteriocin in the case of strain S37, whereas it is ascribable to a nonbacteriocin, likely a reuterin, in the case of strain S42.