Faecal-indicator bacteria and sedimentary processes in estuarine mudflats (Seine, France).

Over a three-year period, quantification of faecal indicators and the molecular detection of Escherichia coli and Salmonella were monitored in sediments from three contrasting mudflats of the Seine estuary (France). The elevation of the mudflat surface was monitored concurrently using a high-resolution altimeter. During the period of the study, estuarine mudflats were areas of deposition for faecal-indicator bacteria and were mainly controlled by sedimentary processes. In the intertidal freshwater and subtidal mudflats, the highest abundances of faecal-indicator bacteria were counted during a depositional period. Maximum levels were observed in the freshwater mudflats during periods of high flow: thermotolerant coliforms: 3.9 x 10(4) cfu cm(-2), enterococci: 1.2 x 10(4) cfu cm(-2), Clostridium perfringens spores: 9.8 x 10(5) spores cm(-2). Loss of culturability of enteric bacteria in sediment microcosms demonstrated the remediatory capacity of the mudflats, even if they might be a secondary source of bacteria-forming spores to the water column through erosion and resuspension events.

Assessment of faecal contamination and the relationship between pathogens and faecal bacterial indicators in an estuarine environment (Seine, France).

The Seine estuary, one of the largest estuaries of the European northwest continental shelf, is subjected to numerous anthropogenic influences. Here we present an assessment of the microbial faecal contamination of the estuary water. The most vulnerable areas were defined on the basis of the fluxes of indicator organisms and the occurrence of Salmonella and Cryptosporidium sp. and Giardia sp. (oo)cysts. The microbial quality of the water changes from upstream to downstream: in the upstream area, contamination by faecal-indicator bacteria and Salmonella occurs during periods of high flow; in the urbanized area, mid-way between the uppermost areas of the estuary and its mouth, discharge from a wastewater treatment plant and a tributary degrade water quality; at the estuary mouth, the accumulation of microorganisms attached to particles in the maximum turbidity zone, particularly Clostridium perfringens spores and oocysts of Cryptosporidium, is accompanied by inputs of ThC and Escherichia coli from tributaries. In some areas, significant strong relations are observed between Salmonella, (oo)cysts of protozoan, and levels of faecal indicators.

Detection of Salmonella in environmental water and sediment by a nested-multiplex polymerase chain reaction assay.

From 1995 to 2002, 53 serovars of Salmonella were isolated in the Seine estuary (France). The 3 serovars most frequently found were S. enterica serovar Typhimurium, S. enterica serovar Infantis and S. enterica serovar Virchow. A nested multiplex PCR (nm-PCR) assay was developed to detect the presence of Salmonella in estuarine water and sediment samples. The target gene used was the phase 1 flagellin fliC chromosomal gene, present in all Salmonella serovars. A set of 4 primers was first used to amplify an 890-bp sequence of the fliC gene, and then a second set of 3 primers was used for the nested PCR. The nmPCR method has been successfully tested for 28 serovars, 13 of which are of epidemiological significance. The detection limit of the assay, without any pre-enrichment step, was estimated at 1 CFU in deionized water, and at 4-5 CFU in the reaction mixture when tested on estuarine water seeded with a Salmonella strain. When the nmPCR was used together with the classical culture method in environmental samples, it gave additional positive results for 11.3% of the sediment samples and 20% of the water samples despite a high background of other bacteria. Overall, the results demonstrated that this molecular approach informed us about the contamination by Salmonella of estuarine water and sediment samples. Positive amplifications suggested the presence of Salmonella DNA and could thus provide information about a recent (culturable) or past (non-culturable, released DNA) contamination of environmental samples by this pathogenic bacteria.