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Background: Lactobacillus species produce biosurfactants that can contribute to the bacteria's ability to prevent microbial infections associated with urogenital and gastrointestinal tracts and the skin. Here, we described the biological and physicochemical properties of biosurfactants produced by Lactobacillus jensenii P 6A and Lactobacillus gasseri P 65 ., Results: The biosurfactants produced by L. jensenii P 6A and L. gasseri P 65 reduced the water surface tension from 72 to 43.2 mN m -1 and 42.5 mN m -1 as their concentration increased up to the critical micelle concentration (CMC) values of 7.1 and 8.58 mg mL -1 , respectively. Maximum emulsifying activity was obtained at concentrations of 1 and 5 mg mL -1 for the P 6A and P 65 strains, respectively. The Fourier transform infrared spectroscopy data revealed that the biomolecules consist of a mixture of carbohydrates, lipids and proteins. The gas chromatography-mass spectrum analysis of L. jensenii P 6A biosurfactant showed a major peak for 14-methypentadecanoic acid, which was the main fatty acid present in the biomolecule; conversely, eicosanoic acid dominated the biosurfactant produced by L. gasseri P 65 . Although both biosurfactants contain different percentages of the sugars galactose, glucose and ribose; rhamnose was only detected in the biomolecule produced by L. jensenii P 6A . Emulsifying activities were stable after a 60-min incubation at 100 °C, at pH 2-10, and after the addition of potassium chloride and sodium bicarbonate, but not in the presence of sodium chloride. The biomolecules showed antimicrobial activity against clinical isolates of Escherichia coli and Candida albicans, with MIC values of 16 µg mL -1 , and against Staphylococcus saprophyticus, Enterobacter aerogenes and Klebsiella pneumoniae at 128 µg mL -1 . The biosurfactants also disrupted preformed biofilms of microorganisms at varying concentrations, being more efficient against E. aerogenes (64%) (P 6A biosurfactant), and E. coli (46.4%) and S. saprophyticus (39%) (P 65 biosurfactant). Both strains of lactobacilli could also co-aggregate pathogens., Conclusions: This report presents the first characterization of biosurfactants produced by L. jensenii P 6A and L. gasseri P 65 . The antimicrobial properties and stability of these biomolecules indicate their potential use as alternative antimicrobial agents in the medical field for applications against pathogens that are responsible for infections in the gastrointestinal and urogenital tracts and the skin.

A bacteriocinogenic Lactobacillus rhamnosus L156.4 strain isolated from the feces of NIH mice was identified by 16S rRNA gene sequencing and MALDI-TOF mass spectrometry. The entire genome was sequenced using Illumina, annotated in the PGAAP, and RAST servers, and deposited. Conserved genes associated with bacteriocin synthesis were predicted using BAGEL3, leading to the identification of an open reading frame (ORF) that shows homology with the L. rhamnosus GG (ATCC 53103) prebacteriocin gene. The encoded protein contains a conserved protein motif associated a structural gene of the Enterocin A superfamily. We found ORFs related to the prebacteriocin, immunity protein, ABC transporter proteins, and regulatory genes with 100% identity to those of L. rhamnosus HN001. In this study, we provide evidence of a putative bacteriocin produced by L. rhamnosus L156.4 that was further confirmed by in vitro assays. The antibacterial activity of the substances produced by this strain was evaluated using the deferred agar-spot and spot-on-the lawn assays, and a wide antimicrobial activity spectrum against human and foodborne pathogens was observed. The physicochemical characterization of the putative bacteriocin indicated that it was sensitive to proteolytic enzymes, heat stable and maintained its antibacterial activity in a pH ranging from 3 to 9. The activity against Lactobacillus fermentum , which was used as an indicator strain, was detected during bacterial logarithmic growth phase, and a positive correlation was confirmed between bacterial growth and production of the putative bacteriocin. After a partial purification from cell-free supernatant by salt precipitation, the putative bacteriocin migrated as a diffuse band of approximately 1.0-3.0 kDa by SDS-PAGE. Additional studies are being conducted to explore its use in the food industry for controlling bacterial growth and for probiotic applications.

Purpose: Class 1 integrons are among the main vehicles that facilitate the spread of antibiotic-resistance genes, with serious public health consequences. The aim of this cross-sectional study was to investigate the presence of class 1 integrons and to characterize their variable regions, as well as the antimicrobial resistance profiles and phylogenetic groups of a collection of Escherichia coli isolates recovered from healthy subjects (n=42) and those with urinary infection (n=40)., Methodology: The methods used included PCR, sequencing and antimicrobial susceptibility testing., Results: PCR screening for the integrase gene (intI1) revealed a higher incidence of class 1 integrons in uropathogenic E. coli (65 %, UPEC) than in commensal isolates (11.9 %). Eight of 31 intI1-positive isolates, all of them UPEC, harboured empty integrons. The variable regions of the other 23 contained gene cassettes encoding resistance to β-lactams (blaOXA-1), aminoglycosides (aadA1 and aadA5), trimethoprim (dfrA1 and dfrA17) and an ORF. To our knowledge this is the first report of an ORF identified as a putative phage tail protein associated with a class 1 integron. The aadA1 and dfrA17-addA5 arrays prevailed in commensal E. coli and UPEC, respectively. UPEC isolates were highly resistant to the antimicrobials tested, in contrast to commensal isolates. The E. coli isolates carrying gene cassettes associated with class 1 integrons were found to be unrelated to any phylogroup or multiresistance., Conclusion: Co-resistance to clinically relevant fluoroquinolone and trimethoprim-sulfamethazole in all UPEC isolates is a cause for concern. These results expand the current knowledge of gene cassettes in both commensal and pathogenic E. coli.

Wastewater treatment plants (WWTPs) harbor bacteria and antimicrobial resistance genes, favoring gene exchange events and resistance dissemination. Here, a culture-based and metagenomic survey of qnrA, qnrB, qnrS, and aac(6')-Ib genes from raw sewage (RS) and activated sludge (AS) of a full-scale municipal WWTP was performed. A total of 96 bacterial isolates were recovered from nalidixic acid-enrichment cultures. Bacteria harboring the aac(6')-Ib gene predominated in RS, whereas qnrS-positive isolates were specific to AS. Novel qnrS- and aac(6')-Ib-cr positive species were identified: Morganella morganii, Providencia rettgeri, and Pseudomonas guangdongensis (qnrS), and Alcaligenes faecalis and P. rettgeri (aac(6')-Ib-cr). Analysis of qnrS and aac(6')-Ib sequences from isolates and clone libraries suggested that the diversity of qnrS is wider than that of aac(6')-Ib. A large number of amino acid mutations were observed in the QnrS and AAC(6')-Ib proteins at previously undetected positions, whose structural implications are not clear. An accumulation of mutations at the C72, Q73, L74, A75 and M76 positions of QnrS, and D181 of AAC(6')-Ib might be important for resistance. These findings add significant information on bacteria harboring qnrS and aac(6')-Ib genes, and the presence of novel mutations that may eventually emerge in clinical isolates., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Antagonistic and synergistic substances are important for interactions between micro-organisms associated with human body surfaces, either in healthy or in diseased conditions. In the present study, such compounds produced by Gardnerella vaginalis strains isolated from women with bacterial vaginosis (BV) were detected in vitro and the antagonistic ones were partially characterized. Among 11 G. vaginalis strains tested, all showed antagonistic activity against at least one of the 22 indicator bacteria assayed. Interestingly, for some of these strains, antagonism reverted to synergism, favouring one of the indicator strains (Peptostreptococcus anaerobius) when the growth medium was changed. Partial characterization of antagonistic substances suggested a bacteriocin-like chemical nature. Depending on growth conditions, G. vaginalis isolated from women with BV produced antagonistic or synergistic compounds for other bacterial components of the vaginal ecosystem. This is the first report to our knowledge of the production of antagonistic and/or synergistic substances by G. vaginalis. This ability may be a pivotal factor in understanding BV and the ecological role of this bacterium in the vaginal environment.

Previous results in the laboratory of the authors showed that Saccharomyces cerevisiae strain 905, isolated during 'cachaça' production, was able to colonize and survive in the gastrointestinal tract of germ-free and conventional mice, and to protect these animals against oral challenge with Salmonella enterica serotype Typhimurium or Clostridium difficile. In the present work, the effects of S. cerevisiae 905 on the translocation of Salm. Typhimurium (mesenteric lymph nodes, Peyer's patches, spleen, liver) as well as on the immune system (number of Küpffer cells, immunoglobulin production, clearance of Escherichia coli B41) were evaluated in gnotobiotic and/or conventional mice. The treatment with the yeast reduced significantly the translocation of Salm. Typhimurium to liver in gnotobiotic animals and to all the organs tested in conventional mice. The number of Küpffer cells per 100 hepatocytes in liver was significantly higher (P<0.05) in yeast mono-associated mice (52.9+/-15.7) than in germ-free controls (38.1+/-9.0). Probably as a consequence, clearance of E. coli B41 from the bloodstream was more efficient in yeast mono-associated animals when compared to germ-free mice. Higher levels (P<0.05) of secretory IgA in intestinal content and of IgA and IgM in serum were observed in yeast mono-associated mice when compared to germ-free group. Concluding, the protection against pathogenic bacteria observed in a previous study was probably due to a modulation of both local and systemic immunity of mice treated with S. cerevisiae 905.