Phages of Bacteroides (GB-124): a novel tool for viral waterborne disease control?

Current fecal indicator bacteria (FIB) and emerging microbial source tracking (MST) methods may indicate the presence and even the likely source of water contamination, but they are less effective at determining the potential risk to health from human enteric viruses. This paper investigates the presence of human-specific phages (detected using a low-cost MST method) in municipal wastewaters (MW) and assesses whether they may be used effectively to screen for the likely presence of human adenovirus (HAdV) and norovirus (NoV). The findings demonstrated that all samples positive for HAdV and/or NoV also contained phages infecting Bacteroides GB-124 (mean = 4.36 log(10) PFU/100 mL) and that GB-124 phages, HAdV, and NoV were absent from samples of nonhuman origin. HAdV and NoV were detected more frequently in MW samples containing higher levels of phages (e.g., >10(2)) and FIB (e.g., >10(3)). Interestingly, at one sewage treatment works (STW), the levels of GB-124 phages present in treated MW were not significantly lower (p = 0.001) than those in untreated MW. There was a positive correlation (R = 0.42) between the size of STW and the number of GB-124 phages present in the final treated effluent. Therefore, the detection of GB-124 phages by a simple phage-lysis method may have considerable potential as a low-cost surrogate for the detection of certain human pathogenic viruses in MW and receiving waters.

Comparison of fifteen commercial assays for detecting Varicella Zoster virus IgG with reference to a time resolved fluorescence immunoassay (TRFIA) and the performance of two commercial assays for screening sera from immunocompromised individuals.

The performance of fifteen, commercially available, VZV IgG assays and an "in house" indirect immunofluorescence (IF) assay has been compared to a reference VZV IgG time resolved immunofluorescence assay (VZV TRFIA). A panel of 273 VZV TRFIA IgG positive serum samples and 136 VZV TRFIA IgG susceptible sera, collected from a number of UK hospitals was used. Irrespective of the interpretation of equivocal results the most sensitive assays were Dade Behring EIA (97.4%), "in house" IF (95.2%), Human EIA (92.3%) and Becton Dickinson latex agglutination (94.1%). The least sensitive assays were Virion EIA (69.6%), Diesse EIA (68.9%) and Diasys EIA (68.5%). The least sensitive (<70%) assays all had >99.0% specificity whereas the most sensitive assays had lower specificities; for example, Dade Behring EIA had a specificity of 69.9% when equivocals were treated as VZV IgG negative. For some assays e.g. Dade Behring EIA there were major discrepancies between our findings and those reported by the manufacturer which may reflect the constitution of the panel(s) of sera used for evaluation or the reference method adopted or the choice of cut-off criteria (particularly relevant to our findings for the Behring EIA). Care must be taken to choose an assay with high specificity in order to accurately assess the need for vaccination or immunoprophylaxis; however, high sensitivity is preferable to prevent inappropriate and expensive treatment.

Application of the heteroduplex mobility assay (HMA) for the investigation of the genomic diversity among noroviruses in environmental samples.

Recent studies have demonstrated the widespread contamination of river and seawater with noroviruses (NV), often with more than one strain. The heteroduplex mobility assay (HMA) in which amplicons from study samples are hybridised (by denaturing and reannealing) to amplicons from reference strains and resolved by electrophoresis, has the potential to provide a simple and rapid means to identify samples containing multiple NV strains and to establish the diversity of strains within that sample. PCR amplicons from environmental samples that were tested directly in the HMA assay were shown to contain more than one strain. In order to evaluate HMA for investigations of NV diversity in environmental samples, amplicons from three representative samples were cloned and, for each, 20 amplicons derived from individual clones were analysed by HMA. Between two and six different HMA profiles were demonstrated among clones from a single sample indicating the extent of NV diversity in the sample. Sequence analysis confirmed the relationship of HMA profile and NV 'genotype'. Far greater diversity was seen among Genogroup (G) II (Ni/E3) amplicons than Genogroup (G) I (Ando/E3) amplicons (generated from the RNA dependent RNA polymerase region of the ORF1 of noroviruses), which often contained only a single strain, which is reflective of the greater prevalence of GII NVs over GI NVs. Overall, four GII and four GI strains were identified in these environmental water/sewage samples.

Microbial contamination of two urban sandstone aquifers in the UK.

Development of urban groundwater has historically been constrained by concerns about its quality. Rising urban water tables and overabstraction from rural aquifers in the UK have led to a renewed interest in urban groundwater, particularly the possibility of finding water of acceptable quality at depth. This study assessed the microbial quality of groundwater collected from depth-specific intervals over a 15-month period within the Permo-Triassic Sherwood Sandstone aquifers underlying the cities of Nottingham and Birmingham. Sewage-derived bacteria (thermotolerant coliforms, faecal streptococci and sulphite-reducing clostridia) and viruses (enteroviruses, Norwalk-like viruses, coliphage) were regularly detected to depths of 60 m in the unconfined sandstone and to a depth of 91 m in the confined sandstone. Microbial concentrations varied temporally and spatially but increased frequency of contamination with depth coincided with geological heterogeneities such as fissures and mudstone bands. Significantly, detection of Norwalk-like viruses and Coxsackievirus B4 in groundwater corresponded with seasonal variations in virus discharge to the sewer system. The observation of low levels of sewage-derived microbial contaminants at depth in the Triassic Sandstone aquifer is explained by the movement of infinitesimal proportions of bulk (macroscopic) groundwater flow along preferential pathways (e.g., fissures, bedding planes). The existence of very high microbial populations at source (raw sewage) and their extremely low detection limits at the receptor (multilevel piezometer) enable these statistically extreme (microscopic) flows to be traced. Rapid penetration of microbial contaminants into sandstone aquifers, not previously reported, highlights the vulnerability of sandstone aquifers to microbial contamination.

Virus hazards from food, water and other contaminated environments.

Numerous viruses of human or animal origin can spread in the environment and infect people via water and food, mostly through ingestion and occasionally through skin contact. These viruses are released into the environment by various routes including water run-offs and aerosols. Furthermore, zoonotic viruses may infect humans exposed to contaminated surface waters. Foodstuffs of animal origin can be contaminated, and their consumption may cause human infection if the viruses are not inactivated during food processing. Molecular epidemiology and surveillance of environmental samples are necessary to elucidate the public health hazards associated with exposure to environmental viruses. Whereas monitoring of viral nucleic acids by PCR methods is relatively straightforward and well documented, detection of infectious virus particles is technically more demanding and not always possible (e.g. human norovirus or hepatitis E virus). The human pathogenic viruses that are most relevant in this context are nonenveloped and belong to the families of the Caliciviridae, Adenoviridae, Hepeviridae, Picornaviridae and Reoviridae. Sampling methods and strategies, first-choice detection methods and evaluation criteria are reviewed.

Surveillance of adenoviruses and noroviruses in European recreational waters.

Exposure to human pathogenic viruses in recreational waters has been shown to cause disease outbreaks. In the context of Article 14 of the revised European Bathing Waters Directive 2006/7/EC (rBWD, CEU, 2006) a Europe-wide surveillance study was carried out to determine the frequency of occurrence of two human enteric viruses in recreational waters. Adenoviruses were selected based on their near-universal shedding and environmental survival, and noroviruses (NoV) selected as being the most prevalent gastroenteritis agent worldwide. Concentration of marine and freshwater samples was done by adsorption/elution followed by molecular detection by (RT)-PCR. Out of 1410 samples, 553 (39.2%) were positive for one or more of the target viruses. Adenoviruses, detected in 36.4% of samples, were more prevalent than noroviruses (9.4%), with 3.5% GI and 6.2% GII, some samples being positive for both GI and GII. Of 513 human adenovirus-positive samples, 63 (12.3%) were also norovirus-positive, whereas 69 (7.7%) norovirus-positive samples were adenovirus-negative. More freshwater samples than marine water samples were virus-positive. Out of a small selection of samples tested for adenovirus infectivity, approximately one-quarter were positive. Sixty percent of 132 nested-PCR adenovirus-positive samples analysed by quantitative PCR gave a mean value of over 3000 genome copies per L of water. The simultaneous detection of infectious adenovirus and of adenovirus and NoV by (RT)PCR suggests that the presence of infectious viruses in recreational waters may constitute a public health risk upon exposure. These studies support the case for considering adenoviruses as an indicator of bathing water quality.