Reduction of bacterial biofilm formation using marine natural antimicrobial peptides.

There is an important need for the development of new "environmentally-friendly" antifouling molecules to replace toxic chemicals actually used to fight against marine biofouling. Marine biomass is a promising source of non-toxic antifouling products such as natural antimicrobial peptides produced by marine organisms. The aim of this study was to demonstrate the efficiency of antimicrobial peptides extracted from snow crab (SCAMPs) to reduce the formation of marine biofilms on immerged mild steel surfaces. Five antimicrobial peptides were found in the snow crab hydrolysate fraction used in this study. SCAMPs were demonstrated to interact with natural organic matter (NOM) during the formation of the conditioning film and to limit the marine biofilm development in terms of viability and bacterial structure. Natural SCAMPs could be considered as a potential alternative and non-toxic product to reduce biofouling, and as a consequence microbial induced corrosion on immerged surfaces.

Impact of polymer-coated silver nanoparticles on marine microbial communities: a microcosm study.

The use of silver nanoparticles (AgNPs) in consumer products is increasing drastically and their potential environmental impacts on aquatic organisms from bacterial communities to vertebrates are not well understood. This study reports on changes in marine bacterial richness using denaturing gradient gel electrophoresis (DGGE), and overall community abundance determined by flow cytometry in marine microcosms exposed to polymer-coated AgNPs (20±5 nm) and ionic silver (Ag(+)). Our study clearly demonstrated that at low concentrations (5 and 50 µg L(-1) total silver), un-aggregated polymer-coated AgNPs and dissolved Ag(+) contamination produced similar effects: a longer lag phase suggesting an adaptation period for microorganisms. As richness decreased in the treated samples, this longer lag phase could correspond to the selection of a fraction of the initial community that is insensitive to silver contamination. Polymer-coated AgNPs preserved their bactericidal properties even under the high ionic strength of estuarine waters.

Optimization of microbial DNA extraction and purification from raw wastewater samples for downstream pathogen detection by microarrays.

Numerous waterborne pathogens are difficult to detect and enumerate with accuracy due to methodological limitations and high costs of direct culturing. The purity of DNA extracted from wastewater samples is an important issue in the sensitivity and the usefulness of molecular methods such as polymerase chain reaction (PCR) and hybridizations on DNA microarrays. Ten different DNA extraction procedures, including physical and chemical extraction and purification steps, were examined to ascertain their relative effectiveness for extracting bacterial DNA from wastewater samples. The quality of the differentially extracted DNAs was subsequently assessed by PCR amplification and microarray hybridization. Our results showed that great differences existed among the ten procedures and only a few of the methods gave satisfactory results when applied to bacterial pathogens. This observation suggested that the extraction method needed to be carefully selected to produce significant and confident results in the detection of pathogens from environmental samples.

Evaluation of ChemChrome V6 for bacterial viability assessment in waters.

The efficiency of ChemChrome B (CB) and ChemChrome V6 (CV6) dyes to stain viable bacterial cells in water was compared. Both dyes are fluorogenic esters converted to free fluorescein by esterase activity. The dyes were applied to a wide variety of bacterial species, including those poorly stained by CB, and to natural waters. Some species tested gave unacceptable low fluorescence intensities by being inefficiently or non-labelled with the CB. In contrast, CV6-stained bacteria were easily detected by both flow cytometry and solid-phase cytometry. As a consequence, higher viable cell counts were found with CV6 compared with CB in natural waters. Viable counts determined by CV6 staining were always higher than cfu counts. In contrast, respiring cell counts (CTC) were always lower than CV6 counts and, in the case of tap and mineral waters, they were lower than cfu counts.

Diversity of Salmonella strains isolated from the aquatic environment as determined by serotyping and amplification of the ribosomal DNA spacer regions.

Salmonella species are pathogenic bacteria often detected in sewage, freshwater, marine coastal water, and groundwater. Salmonella spp. can survive for long periods in natural waters, and the persistence of specific and epidemic strains is of great concern in public health. However, the diversity of species found in the natural environment remains unknown. The aim of this study was to investigate the diversity of Salmonella strains isolated from different natural aquatic systems within a Mediterranean coastal watershed (river, wastewater, and marine coastal areas). A total of 574 strains isolated from these natural environments were identified by both conventional serotyping and the ribosomal spacer-heteroduplex polymorphism (RS-HP) method (M. A. Jensen and N. Straus, PCR Methods Appl. 3:186-194, 1993). More than 40 different serotypes were found, and some serotypes probably mobilized from widespread animal-rearing activities were detected only during storm events. These serotypes may be good indicators of specific contamination sources. Furthermore, the RS-HP method based on the PCR amplification of the intergenic spacer region between the 16S and 23S rRNA genes can produce amplicon profiles allowing the discrimination of species at both serotype and intraserotype levels. This method represents a powerful tool that could be used for rapid typing of Salmonella isolates.

Effectiveness of CSE to counterstain particles and dead bacterial cells with permeabilised membranes: application to viability assessment in waters.

The CSE dye (Chemunex, Maisons-Alfort, France) was combined with an activity marker to improve bacterial activity assessment in natural waters. Its effectiveness to counterstain dead cells with permeabilised membranes was investigated on live and dead cells of a variety of strains from collections or isolated from the natural environment. Cells were killed by heat treatment. For all strains tested, the fluorescent dye showed an intense staining of killed cells having permeabilised membranes while no significant signal was detected when applied to live cells. Furthermore, the CSE dye had no toxicity on viable cells. Then, CSE was combined with the ChemChrome V6 dye (Chemunex) to assess the activity of bacterial cells in different waters. Both fluorescences were analysed simultaneously by solid-phase cytometry. The active cell counts were sometimes lower when both dyes were combined suggesting that CSE was able to counterstain cells having a residual esterase activity and compromised membranes. These cells were subtracted from the active cell counts determined with ChemChrome V6. In most samples, active cell counts were congruent with those determined by the direct viable count method.