Massive outbreak of viral gastroenteritis associated with consumption of municipal drinking water in a European capital city.

On 24 August 2008, an outbreak alert regarding cases of acute gastroenteritis in Podgorica triggered investigations to guide control measures. From 23 August to 7 September, 1699 cases were reported in Podgorica (population 136 000) and we estimated the total size of the outbreak to be 10 000-15 000 corresponding to an attack rate of approximately 10%. We conducted an age- and neighbourhood-matched case-control study, microbiologically analysed faecal and municipal water samples and assessed the water distribution system. All cases (83/83) and 90% (80/90) [corrected] of controls drank unboiled chlorinated municipal water [matched odds ratio (mOR) 11.2, 95% confidence interval (CI), 1.6-infinity]. Consumption of bottled water was inversely associated with illness (mOR 0.3, 95% CI 0.1-0.8). Analyses of faecal samples identified six norovirus genotypes (21/38 samples) and occasionally other viruses. Multiple defects in the water distribution system were noted. These results suggest that the outbreak was caused by faecally contaminated municipal water. It is unusual to have such a large outbreak in a European city especially when the municipal water supply is chlorinated. Therefore, it is important to establish effective multiple-barrier water-treatment systems whenever possible, but even with an established chlorinated supply, sustained vigilance is central to public health.

Development of a viral concentration method for bottled water stored in hydrophobic support.

Several protocols have been described for the detection of genomes of enteric viruses from water using two-step procedures: membrane filtration and RT-PCR detection. However, these methods, when applied to bottled water, generally consider only the aqueous phase. Such procedures do not take into account the adhesion of viruses onto the hydrophobic container. Potential adhesion results in loss of viral concentration in the aqueous phase and consequently viral pollution is underestimated in such a system. A procedure based on the addition of surfactant to elute viruses followed by membrane concentration was developed to avoid this underestimation. Firstly, using poliovirus 1 as a model, this study demonstrated that the best solution to recover virus and/or viral genome is a mix of sodium dodecyl sulphate, a nonionic detergent and guanidine thiocyanate. Furthermore, temperature has a significant but low effect on elution. A positively charged 0.2 microm inorganic membrane composed of Alumina (Anodisc, Whatman) is also the best membrane to concentrate viral material before the detection by RT-PCR. Finally, the developed protocol gives significantly higher poliovirus 1 recovery rate than a reference protocol previously described (aqueous phase concentration on Zetapore). The difference can be explained by the recovery of the viruses adsorbed onto the water container.

Survival of infectious Poliovirus-1 in river water compared to the persistence of somatic coliphages, thermotolerant coliforms and Poliovirus-1 genome.

The microbiological quality of water is currently assessed by search for fecal bacteria indicators. There is, however, a body of knowledge demonstrating that bacterial indicators are less resistant to environmental factors than human pathogenic viruses and therefore underestimate the viral risk. As river water is often used as a resource for drinking water production, it is particularly important to obtain a valid estimation of the health hazard, including specific viral risk. This work was conducted to compare the survival of infectious Poliovirus-1 used as a pathogenic virus model to the persistence of, on the one hand, thermotolerant coliforms commonly used as indicators and on the other hand, to somatic coliphages and Poliovirus-1 genome considered as potential indicators. We studied the behavior of infectious Poliovirus-1 and the three (potential) indicators of viral contamination in river water at three different temperatures (4 degrees C,18 degrees C and 25 degrees C). This experiment was performed twice with river water sampled at two different periods, once in winter and once in summer. Our results showed that the survival of thermotolerant coliforms can be 1.5-fold lower than infectious Poliovirus-1. In contrast, under all our experimental conditions, somatic coliphages and Poliovirus-1 genome persisted longer than infectious Poliovirus-1, surviving, respectively, 2-6-fold and about 2-fold longer than infectious Poliovirus-1. According to our results exclusively based on survival capacity, somatic coliphages and viral genome, unlike thermotolerant coliforms appear to be better indicators of viral contamination in river water. Moreover, the disappearance of viral genome is well-correlated to that one of infectious virus irrespective of the conditions tested.

Recovery of feline calicivirus infectious particles and genome from water: comparison of two concentration techniques.

The aim of this work was to determine the recovery rate of feline calicivirus (FCV-F9) infectious particles and genome from water after a concentration step using either adsorption elution on glass wool or filtration through an electropositive membrane. The results showed that the membrane filtration technique allowed a 75% recovery rate of FCV-F9 infectious particles while the yield was only 5.3% for FCV-F9 genome. Using the glass wool adsorption-elution technique, the recovery rate of FCV-F9 infectious particles was 0.5% while the yield was 102.5% with Poliovirus 1.

Comparison of coliforms and coliphages as tools for assessment of viral contamination in river water.

The aim of the study was to evaluate the presence of pathogenic viruses in the Moselle River and to compare the usefulness of thermotolerant coliforms and somatic coliphages as tools for river water quality assessment in terms of viral contamination. Thermotolerant coliforms and somatic coliphages were enumerated by standardized methods in 170 samples of river water drawn from five sampling sites along the Moselle River (eastern France). BGM cell culture and integrated cell culture-reverse transcription-PCR DNA enzyme immunoassay were used to determine the presence of pathogenic viral genome (Enterovirus and Norovirus genogroup II [GGII]) and infectious Enterovirus spp. in 90 1-liter samples. No infectious Enterovirus spp. were isolated, but Enterovirus and Norovirus GGII genomes were detected in 38% of the samples. Norovirus GGII genome was mostly detected in winter, whereas Enterovirus genome was mostly detected in summer and fall. Somatic coliphages appeared to be less sensitive to higher river water temperature than thermotolerant coliforms. Furthermore, the number of river water samples positive for pathogenic viral genome increased with increasing concentration of somatic coliphages, whereas coliform concentration was unrelated to viral genome contamination. Consequently somatic coliphages, which are less sensitive to environmental factors than thermotolerant coliforms in river water, would provide a promising tool for assessment of river water quality in terms of fecal and viral pollution.