Sewage impact on shellfish microbial contamination.

Coastal areas are frequently contaminated by microorganisms of human origin, due to high population density and low seawater renewal. To evaluate the impact of wastewater input on shellfish quality, a study was conducted in Brittany (France) over a period of 20 months. A hydrodynamic model was used to simulate wastewater impact on microbial water quality. To validate the model, wastewater from the three main sewage treatment plants and shellfish from three sites were sampled monthly. Bacterial indicators (E. coli), F-RNA phages were searched for by culture and noroviruses by RT-PCR and hybridisation. These microorganisms were detected in the three effluents and clams, with no marked seasonal variation. The microbial concentrations in the two oyster beds, distant from the effluent outfall, were low, and only three of the samples were positive for norovirus. For simulation, the winter wastewater inputs of E. coli and phages were calculated and an estimation for norovirus flux was made from the epidemic situation in the population. The microbial behaviour was included in the model by a decay-rate factor. Results from the model calculations were found to be very similar to E. coli and phage concentrations observed in shellfish. For noroviruses, the model indicated that shellfish distant from the wastewater input were under the detection limit of the RT-PCR method. This study demonstrated the use of modelisation to interpret norovirus contamination in various areas.

Human enteric viruses in Coquina clams associated with a large hepatitis A outbreak.

An outbreak of hepatitis A, affecting 183 people, occurred in Valencia (Spain). Epidemiological evidence pointed to an association of the outbreak with consumption of Coquina clams (Donax sp), imported frozen from Peru. Shellfish were analysed for the presence of hepatitis A virus (HAV), enteroviruses, rotaviruses, astroviruses, caliciviruses and hepatitis E virus. HAV was detected in 75% of assayed shellfish samples. Other enteric viruses were occasionally found in the same samples. Molecular epidemiological analysis of fragments of the VP1/2A and the 5' end of the genome from shellfish and sera isolates, revealed the presence of six variants belonging to a single genotype.

[Magnitude of rainfall on viral contamination of the marine environment during gastroenteritis epidemics in human coastal population]. / Importance de la pluviométrie sur la contamination virale du milieu littoral lors de phénomènes épidémiques dans la population.

BACKGROUND: Sewage treatments are not efficient to eliminate enteric microorganisms. Viruses are able to persist and are discharged into the marine environment with treated effluents. Few data are now available on the magnitude and the contributive processes of marine viral contamination. This work evaluates the relationship between the magnitude of rainfall and the viral contamination of the marine environment during winter epidemics of gastroenteritis in human coastal populations. METHODS: A RT-PCR method was used to detect enterovirus, hepatitis A virus, Norwalk-like virus, astrovirus and rotavirus in shellfish, harvested monthly between August 1995 and July 1998. The frequency of virus detection in shellfish was expressed as an Index of Viral Contamination. Acute gastroenteritis in the population was estimated using the French Sentinel System for Monitoring of Communicable Diseases. Rainfall effects on the efficiency of sewage treatment were assessed using an estimated staying time of sewage effluents in the plant. RESULTS: The results indicate that the highest viral contamination occurs in winter. Maximal indexes of viral contamination were respectively 70% in January 1996, 100% in January 1997, but only 31% in January 1998. Viral contamination variations seemed to follow the pattern of the winter epidemic of acute gastroenteritis in the local population in 1996 and 1997. These observations should be linked to the winter rainfalls. Heavy rains on short periods of time could create an hydraulic overload in the sewage treatment plant, reducing the staying time of the sewage effluents and thus the efficiency of the disinfection process. CONCLUSION: The magnitude of the viral contamination of shellfish seems to result from the simultaneity between the winter epidemics of acute gastroenteritis in the coastal population and heavy rainfall. To prevent public health hazards associated with shellfish consumption, the monitoring of microbiological quality in shellfish harvesting areas should include accompagning survey of viral epidemic in the coastal population, and of sewage outputs in the coastal environment.

Three-year study to assess human enteric viruses in shellfish.

The main pathogenic enteric viruses able to persist in the environment, such as hepatitis A virus (HAV), Norwalk-like virus (NLV), enterovirus (EV), rotavirus (RV), and astrovirus (AV), were detected by reverse transcription-PCR and hybridization in shellfish during a 3-year study. Oyster samples (n = 108), occasionally containing bacteria, were less frequently contaminated, showing positivity for AV (17%), NLV (23%), EV (19%), and RV (27%), whereas mussel samples, collected in areas routinely impacted by human sewage, were more highly contaminated: AV (50%), HAV (13%), NLV (35%), EV (45%), and RV (52%). Sequences obtained from HAV and NLV amplicons showed a great variety of strains, especially for NLV (strains close to Mexico, Snow Mountain Agent, or Norwalk virus). Viral contamination was mainly observed during winter months, although there were some seasonal differences among the viruses. This first study of virus detection over a fairly long period of time suggests that routine analysis of shellfish by a molecular technique is feasible.

Molecular epidemiological survey of rotaviruses in sewage by reverse transcriptase seminested PCR and restriction fragment length polymorphism assay.

Rotavirus double-stranded RNA was detected directly in sewage treatment plant samples over a 1-year period by reverse transcription followed by PCR amplification of the VP7 gene and Southern blot hybridization. The presence of naturally occurring rotaviruses was demonstrated in 42% of raw sewage samples and in 67% of treated effluent samples. Amplified viral sequences were analyzed by restriction enzymes. Ten different restriction profiles were characterized, most of which were found in treated effluent samples. A mixture of restriction profiles was observed in 75% of contaminated effluent samples. The profiles were compared with those obtained from human rotavirus isolates involved in infections in children from the same area (six different profiles were detected). Five identical viral sequences were detected in both environmental and clinical samples. Restriction profiles were also compared to profiles from known genomic sequences of human and animal viruses. Both human and animal origins of rotavirus contamination of water seemed likely.