Detection of CTX-M-15 harboring Escherichia coli isolated from wild birds in Tunisia.

BACKGROUND: The spreading of antibiotic resistant bacteria is becoming nowadays an alarming threat to human and animal health. There is increasing evidence showing that wild birds could significantly contribute to the transmission and spreading of drug-resistant bacteria. However, data for antimicrobial resistance in wild birds remain scarce, especially throughout Africa. The aims of this investigation were to analyze the prevalence of ESBL-producing E. coli in faecal samples of wild birds in Tunisia and to characterize the recovered isolates. RESULTS: One hundred and eleven samples were inoculated on MacConkey agar plates supplemented with cefotaxime (2 µg/ml). ESBL-producing E. coli isolates were detected in 12 of 111 faecal samples (10.81%) and one isolate per sample was further characterized. ß-lactamase detected genes were as follows: blaCTX-M-15 (8 isolates), blaCTX-M-15 + blaTEM-1b (4 isolates). The ISEcp1 and orf477 sequences were found respectively in the regions upstream and downstream of all blaCTX-M-15 genes. Seven different plasmid profiles were observed among the isolates. IncF (FII, FIA, FIB) and IncW replicons were identified in 11 CTX-M-15 producing isolates, and mostly, other replicons were also identified: IncHI2, IncA/C, IncP, IncI1 and IncX. All ESBL-producing E. coli isolates were integron positive and possessed "empty" integron structures with no inserted region of DNA. The following detected virulence genes were: (number of isolates in parentheses): fimA (ten); papC (seven); aer (five); eae (one); and papGIII, hly, cnf, and bfp (none). Molecular typing using pulsed-field gel electrophoresis and multilocus sequence typing showed a low genetic heterogeneity among the 12 ESBL-producing strains with five unrelated PFGE types and five different sequence types (STs) respectively. CTX-M-15-producing isolates were ascribed to phylogroup A (eleven isolates) and B2 (one isolate). CONCLUSION: To our knowledge, this study provides the first insight into the contribution of wild birds to the dynamics of ESBL-producing E. coli in Tunisia.

Antimicrobial Activities of Bacteria Associated with the Brown Alga Padina pavonica.

Macroalgae belonging to the genus Padina are known to produce antibacterial compounds that may inhibit growth of human- and animal pathogens. Hitherto, it was unclear whether this antibacterial activity is produced by the macroalga itself or by secondary metabolite producing epiphytic bacteria. Here we report antibacterial activities of epiphytic bacteria isolated from Padina pavonica (Peacocks tail) located on northern coast of Tunisia. Eighteen isolates were obtained in pure culture and tested for antimicrobial activities. Based on the 16S rRNA gene sequences the isolates were closely related to Proteobacteria (12 isolates; 2 Alpha- and 10 Gammaproteobacteria), Firmicutes (4 isolates) and Actinobacteria (2 isolates). The antimicrobial activity was assessed as inhibition of growth of 12 species of pathogenic bacteria (Aeromonas salmonicida, A. hydrophila, Enterobacter xiangfangensis, Enterococcus faecium, Escherichia coli, Micrococcus sp., Salmonella typhimurium, Staphylococcus aureus, Streptococcus sp., Vibrio alginoliticus, V. proteolyticus, V. vulnificus) and one pathogenic yeast (Candida albicans). Among the Firmicutes, isolate P8, which is closely related to Bacillus pumilus, displayed the largest spectrum of growth inhibition of the pathogenic bacteria tested. The results emphasize the potential use of P. pavonica associated antagonistic bacteria as producers of novel antibacterial compounds.

Cationic, anionic and neutral dyes: effects of photosensitizing properties and experimental conditions on the photodynamic inactivation of pathogenic bacteria.

The aim of this study was to evaluate the photobactericidal effect of four photosensitizers (PSs) with different structural and physico-photochemical properties, namely mesotetracationic porphyrin (T4MPyP), dianionic rose Bengal (RB), monocationic methylene blue (MB) and neutral red (NR). Their photokilling activity was tested in vitro on pathogenic bacteria such as Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) suspended in nutrient broth (NB) and in phosphate buffered saline (PBS) through following their influence on the PSs antimicrobial efficacy. Photodynamic inactivation (PDI) experiments were performed using visible light (L) and different PSs concentrations (20-70 µM). The ability of these PSs to mediate bacterial photodynamic inactivation was investigated as a function of type of PS and its concentrations, spectral and physico-chemical properties, bacterial strain, irradiation time and suspending medium. Indeed, they showed antibacterial effects against S. aureus and P. aeruginosa with significant difference in potency. Staphylococcus aureus suspended in NB showed 0.92 log units reduction in viable count in the presence of T4MPyP at 20 µM. Changing the suspending medium from NB to PBS, S. aureus was successfully photoinactivated by T4MPyP (20 µM) when suspended in PBS at least time exposure (10 and 30 min), followed by MB and RB.

The isomerase Rrd1 mediates rapid loss of the Sgs1 helicase in response to rapamycin.

In Saccharomyces cerevisiae , rapamycin exposure inhibits the target of rapamycin (TOR) signaling pathway, causing a profound alteration in the transcription pattern of many genes, including those involved in ribosome biogenesis and nutritional changes. Deletion of the RRD1 gene encoding a peptidyl prolyl isomerase resulted in mutants that are resistant to rapamycin. These rrd1Δ mutants are unable to efficiently downregulate genes such as ribosomal protein genes, or to upregulate genes involved in diauxic shift. It is believed that the isomerase function of Rrd1 plays a role in changing the transcriptional profile upon rapamycin exposure. Herein, we set out to search for genes that when deleted in the rrd1Δ mutant would suppress the rapamycin-resistant phenotype. The analysis revealed that deletion of the SGS1 gene in the rrd1Δ mutant partially suppresses the rapamycin-resistant phenotype of the single rrd1Δ mutant. SGS1 encodes a helicase that functions in many biological processes, including transcriptional regulation. We further show, and for the first time, that Sgs1 is rapidly lost in the parent cells in response to rapamycin, but not by other agents. Interestingly, Sgs1 reduction was completely blocked in the rrd1Δ mutant, suggesting that Rrd1 is required to mediate this process. Genes such as PUT4 and HSP42, known to be upregulated in the parent in response to rapamycin, were not induced in the rrd1Δ mutant if the SGS1 gene was deleted. Since Sgs1 plays a role in transcriptional regulation, we propose that it acts as a repressor of a subset of rapamycin responsive genes. Thus, the observed Rrd1-dependent reduction in Sgs1 level may promote expression of specific classes of genes in response to rapamycin.

The role of reactive oxygen species in Staphylococcus aureus photoinactivation by methylene blue.

Methylene blue (MB) has been shown to photoinduce the direct inactivation of Gram-positive bacteria Staphylococcus aureus (S. aureus) in water. We have investigated the mechanism of S. aureus photoinactivation conducting firstly sodium azide (reactive ion N3-), as a good physical singlet oxygen quencher, then the amino acids tryptophan (Trp), as a non-specific singlet oxygen quencher and the mannitol, as an hydroxyl free radical scavenger. Inactivation of MB photosensitization is the antioxidants type dependent. When the bacteria was treated with MB (20 microM) under light during 10 min of exposure, it was found that survival fraction had decreased dramatically to about 31.27+/-5.39%. The presence of sodium azide and Trp failed to shown any protection from the MB photodynamic activity. In the presence of mannitol, S. aureus could be protected, reaching a protection level of about 27%. It is possible that the photodynamic activity of MB occurred in part, via a Type I mechanism in which *OH was produced.The interactions between MB and S. aureus were studied spectrophotometrically. This demonstrated that a metachromatic reaction took place between MB and S. aureus bacteria. Furthermore, S. aureus bacteria induced additional dimerization of MB.