A study of enteric viruses in raw and treated sewage from two secondary treatment plants, which received sewage from Oslo city (plant A) and small municipalities in Hedmark county in Norway (plant B), showed high levels of noro-, adeno-, and bocavirus throughout the year. A seasonal variation was observed for adeno- and GII norovirus with higher levels during winter and bocavirus that had more positive samples during winter. The virus concentrations in raw sewage were comparable in the two plants, with medians (log10 genome copies per liter) of 6.1, 6.3, 6.0, and 4.5 for noro GI, noro GII, adeno-, and bocavirus, respectively. The level of hepatitis E virus was not determined as it was below the limit of quantification. The mean log10 virus reduction was 0.55 (plant A) and 1.44 (plant B) with the highest reduction found in the plant with longer hydraulic retention time. The adenoviruses were dominantly serotype 41, while serotype 12 appeared sporadically. Of the 102 raw and treated sewage samples that were tested, eight were positive for hepatitis E virus of which four were from treated sewage. Two of the four obtained gene sequences from hepatitis E virus originated from the rural sewage samples and showed high similarity with a genotype 3 strain of hepatitis E virus detected in local piglets. Two other hepatitis E virus sequences obtained from urban sewage samples showed high similarities with genotype 3 strains isolated from urban sewage in Spain and a human genotype 1 isolate from India. The study gives information on the levels of noroviruses in raw and treated sewage, which is valuable to risk assessment, information indicating that some infections with hepatitis E viruses in Norway have a regional origin and that human bocavirus 2 and 3 are prevalent in the Norwegian population.
A long RT-PCR method was developed to amplify the norovirus genome. Starting from RNA extracted directly from clinical samples and using broadly reactive primers, it can generate near full-length amplicons that allow for easy determination of the near complete genomic sequence. Two norovirus isolates from Toronto, Canada, in 2002 and 2005 were sequenced. This approach will facilitate molecular epidemiology studies of noroviruses.
On January 29, 2015, the city of Tampere environmental health officers were informed of a possible foodborne outbreak among customers who had eaten lunch in restaurant X. Employees of electric companies A and B had a sudden onset of gastrointestinal symptoms. We conducted a retrospective cohort study to identify the vehicle, source, and causative agent of the outbreak. A case was defined as an employee of companies A or B with diarrhea and/or vomiting who ate lunch at Restaurant X on January 26, 2015. All employees of the companies attending the implicated lunch were invited to participate in the cohort study. Environmental investigation was conducted. Twenty-one responders were included in statistical analysis, of which 11 met with the case definition. Of the 15 food items consumed by participants, four food items were associated with gastroenteritis. Of four kitchen staff, three tested positive for norovirus GIP7, the strain was found earlier in the community. No patient samples were obtained. Level of hygiene in the kitchen was inadequate. Infected kitchen staff probably transmitted norovirus by inadequate hygiene practices. No new cases associated with Restaurant X were reported after the hygiene practices were improved.
This study investigated the presence of norovirus and adenovirus, especially enteric adenovirus, on the environmental surfaces (n?=?481) and military conscripts' hands (n?=?109) in two Finnish garrisons (A and B) in 2013 and 2014. A questionnaire study was conducted to reveal possible correlations between viral findings on the conscripts' hands and their acute gastroenteritis symptoms. In addition to the swab samples, 14 fecal samples were obtained for viral analysis. In total, norovirus was present in 9.0?% of the surface swabs in 2013, whereas enteric adenovirus was present in 0.0?% and non-enteric adenovirus in 9.4?%. In the same year, 2.6?% of the hand swabs contained norovirus, 2.6?% enteric adenovirus, and 40.3?% non-enteric adenovirus. Norovirus GI.6 was continually detected on the surfaces of garrison A, and identical virus was detected in some of the fecal samples. In garrison B, two slightly different norovirus GII.4 strains were present on the surfaces. The questionnaires revealed no recent acute gastroenteritis cases in garrison A, but in garrison B, where the norovirus-positive hand swabs were collected, 30.6?% of the conscripts reported of recent symptoms. In 2014, norovirus was rarely detected, but adenovirus was again frequently present, both on the surfaces and hands. Taken together, our results suggest that gastroenteritis outbreaks occurred in 2013, but not in 2014. Due to the low number of hand swabs positive for enteric viruses, no conclusions about associations between viral findings and gastroenteritis symptoms could be drawn. This study increased our understanding of the possible transmission of viruses via contaminated environment and hands.
Cites: Int J Food Microbiol. 2006 Apr 15;108(1):84-9116473426
Common blue mussels (Mytilus edulis), horse mussels (Modiolus modiolus), and flat oysters (Ostrea edulis) obtained from various harvesting and commercial production sites along the Norwegian coast were screened for the presence of norovirus by a real-time reverse transcription (RT)-nested PCR assay and for possible indicators of fecal contamination, i.e., for F-specific RNA bacteriophages (F-RNA phages) by plaque assay and for human adenoviruses and human circoviruses by nested PCR assay. The aims were to obtain relevant information for assessing the risk of transmission of enteric viruses by shellfish and to investigate the potential of various indicator viruses in routine screening. Noroviruses were detected in 6.8% of the samples, and the indicators were detected in 23.8% (F-RNA phages), 18.6% (adenoviruses), and 8.0% (circoviruses) of the samples. A seasonal variation was observed, with the exception of circoviruses, with more positive samples in the winter. A positive correlation was found between F-RNA phages and noroviruses. However, F-RNA phages were present in only 43% of the norovirus-positive samples. The results show that mussels from the Norwegian coast can constitute a risk of infection with enteric viruses and that routine testing of samples may be justified. Advantages and disadvantages of various options for screening are discussed.
The public health impact of the emergence of new norovirus (NoV) strains is uncertain. A biennial pattern of alternating quiescent and epidemic levels of NoV outbreak activity associated with the emergence of new GII.4 variants was observed in Alberta, Canada, between July 2000 and June 2008. In this study, NoV genogroup I (GI) and GII strains isolated from 710 outbreak specimens in Alberta between July 2008 and January 2013 were characterized to update historical data. The seasonality and annual variation in NoV outbreak burden were analyzed over a 10-year period (July 2002 to June 2012). We found that GII.4-2006b had persisted as the predominant variant over three observation periods (July 2006 to June 2009) during which the biennial NoV outbreak pattern continued. The emergence of GII.4-2010 (winter 2009) was not associated with increased outbreak activity, and outbreak activity between July 2009 and June 2012 when GII.4-2010 predominated (67.5 to 97.7%) did not follow a biennial pattern. GII.4-2012 first emerged in Alberta in September 2011 and became predominant in observation period July 2012 to June 2013. NoV GI, relatively rare in past years, had a higher activity level (37.3%) as represented by GI.6 and GI.7 in the winter of 2012 to 2013. A higher proportion of GI outbreaks occurred in non-health care facility settings compared to GII. Our study suggests that factors other than new variants emergence contribute to the levels of NoV outbreak activity in Alberta.
Genotyping of noroviruses that had circulated in the territory of Nizhny Novgorod during 6 epidemic seasons (2006 - 2012), detection of dominating genovariants and analysis of their change.
Feces samples from children hospitalized in an intestinal infection department of one of the infectious disease hospitals of Nizhny Novgorod served as material for the study. Noroviruses were detected by reverse transcription polymerase chain reaction. Genotypes and gene variants were determined by analysis of nucleotide sequences of viral genome regions coding capsid protein and RNA-dependent RNA-polymerase.
During examination of 6589 children with an acute intestinal infection between July 2006 and June 2012 noroviruses were detected in 17.55% of cases. Nucleotide sequences of capsid and/or polymerase gene regions were determined for 114 norovirus isolates. Genotyping has shown that noroviruses of 8 various genotypes had circulated in the territory of Nizhny Novgorod--GII.1, GII.2, GII.3, GII.4, GII.6, GII.7, GII. 12, GII.13 with the domination of GII.4 noroviruses for the whole observation period. A dynamic of change of epidemic variants of genotype GII.4 noroviruses that had been accompanied by an increase of frequency of detection of norovirus in children hospitalized with acute intestinal infection similar to global was established. A short-term circulation of GII.4 2006b-NN 2008 norovirus subvariant in spring of 2008 and spread of genotype GII.12 norovirus during 2009, 2010 epidemic season were also shown.
The data obtained give evidence to the necessity of norovirus circulation monitoring with the aim of early detection of novel virus variants that may determine an increase of norovirus infection morbidity.
Detection of caliciviruses requires high mutation tolerance and throughput. The development of a rational simple, single tube reverse transcription-real-time quantitative PCR (QPCR) technique for human noroviruses (NV) is reported here. A dual-probe, triple-primer system (NM system) was used for simultaneous detection and preliminary differentiation of NV genogroups in fecal samples. The design was based on a comprehensive analysis of all 1140 NV sequences available in GenBank. A touch-down amplification protocol improved the frequency of detection. The final QPCR was evaluated with 71 fecal samples from outbreak and sporadic cases in Sweden (1997-2004), all calicivirus-positive by electron microscopy. Up to 56 (79 %) were positive. The method is more rational than NV detection methods described previously, and should be a developmental basis for large-scale routine methods for detection of NV.
A total of 101 food-borne and waterborne outbreaks that were caused by norovirus and that resulted in more than 4,100 cases of illness were reported to the Swedish Institute for Infectious Disease Control from January 2002 to December 2006. Sequence and epidemiological data for isolates from 73 outbreaks were analyzed. In contrast to health care-related outbreaks, no clear seasonality could be observed. Sequence analysis showed a high degree of genetic variation among the noroviruses detected. Genogroup II (GII) viruses were detected in 70% of the outbreaks, and of those strains, strains of GII.4 were the most prevalent and were detected in 25% of all outbreaks. The GII.4 variants detected in global outbreaks in health care settings during 2002, 2004, and 2006 were also found in the food-borne outbreaks. GI strains totally dominated as the cause of water-related (drinking and recreational water) outbreaks and were found in 12 of 13 outbreaks. In 14 outbreaks, there were discrepancies among the polymerase and capsid genotype results. In four outbreaks, the polymerase of the recombinant GII.b virus occurred together with the GII.1 or GII.3 capsids, while the GII.7 polymerase occurred together with the GII.6 and GII.7 capsids. Mixed infections were observed in six outbreaks; four of these were due to contaminated water, and two were due to imported frozen berries. Contaminated food and water serve as important reservoirs for noroviruses. The high degree of genetic diversity found among norovirus strains causing food-borne and waterborne infections stresses the importance of the use of broad reaction detection methods when such outbreaks are investigated.
A total of 2312 children admitted to the Nizhni Novgorod infectious disease hospitals for diagnosed acute gastroenteritis were examined using the RT-PCR. The detection rate of human caliciviruses was found to be 6.5% (range 1.2 to 11.2%). The population of calicivuruses presented with noroviruses genogroups I (16%) and II (81%), and sapoviruses (3%). Noroviruses genogroup II included genotypes GII.2 and GII.4 with a preponderance of the variant CII.4-2006b. The detected sapoviruses were clustered with sapoviruses GI.1, GI.2, and GII.1 by phylogenetic analysis.