NoV GI Research Articles

Norovirus risk at a golf course irrigated with reclaimed water: Should QMRA doses be adjusted for infectiousness?

About 25 golf courses in Ontario, Canada have environmental compliance approvals to use reclaimed water for irrigation, where disinfection is confirmed through E. coli limits. A previous study at five Ontario municipal wastewater treatment plants (WWTPs) confirmed that enteric viruses are less susceptible to disinfection than E. coli, when plants provided conventional (secondary or tertiary) treatment and routine (chlorine or UV) disinfection. Here we query whether these four treatment-disinfection scenarios plus 60-day lagoon storage of disinfected effluent would be sufficient to reduce norovirus genogroups I and II (NoV GI and GII) risk of infection to tolerable levels for a golfer who incidentally ingests NoV after handling wet golf balls at a golf course irrigated with reclaimed water. We used our RT-qPCR NoV enumeration datasets from the four treatment-disinfection scenarios above and modeled detected and non-detected NoV by Bayesian inference in ‘R’. Monte Carlo simulation included pre-disinfection NoV GI and GII gene copy densities; Ontario WWTP-derived chlorine and UV log10 reductions; literature-derived effluent storage decay parameters and golfer ingested volumes, followed by six different NoV dose-response (DR) models. Quantitative Microbial Risk Assessment (QMRA) results suggest that there is an unacceptable NoV infection risk when using the conservative assumption that all detected NoV particles (RT-qPCR gene copies) are infectious, in both aggregated or disaggregated form. However, after adjusting for PCR target sequences and for infectiousness using data from recently published studies on cultivation of human NoV in human intestinal enteroids, we noted a significant reduction of infection risk. However, this less conservative (i.e., less protective) assumption for water reuse applications such as golf course irrigation may not be corroborated until human NoV are efficiently and routinely grown in cell cultures. In addition, further studies on drivers of NoV risk estimation by DR models are needed, e.g., the extent of NoV particle aggregation resulting from wastewater treatment, as well as the role of immunity. Meantime, regulatory agencies could consider more stringent treatment-disinfection requirements that target enteric viruses rather than E. coli and testing of actual reclaimed irrigation waters.

Genetic diversity of norovirus genogroup I, II, IV and sapovirus in environmental water in Thailand

BackgroundEnteric caliciviruses, including noroviruses (NoVs) and sapoviruses (SaVs) are the most significant pathogens associated with waterborne and foodborne outbreaks of nonbacterial acute gastroenteritis in humans worldwide. MethodsIn this study, 126 environmental water samples collected from 6 different sources in Chiang Mai, Thailand from November 2016 to July 2018 were examined for the presence of genogroups I, II, IV (GI, GII, GIV) NoVs and SaVs by using RT-nested PCR assays, genome sequencing, and phylogenetic analysis, ResultsForty out of 126 (31.7%) water samples were positive for one or more caliciviruses throughout the years of study with high prevalence in winter. Among 126 tested specimens, 34 (27.0%), 30 (23.8%), 3 (2.4%), and 2 (1.6%) were positive for NoV GI, GII, GIV, and SaV, respectively. For NoV GI, 6 different genotypes were identified with the most predominant of GI.1 genotype (17 strains). In addition, 6 different genotypes of GII were detected with high prevalence of GII.17 (12 strains) and GII.2 (11 strains). It was interesting to note that our study reported the detection of NoV GIV for the first time in water samples in Thailand, and all were GIV.1 genotype. For SaV detection, only 2 water samples were positive for SaV GI. ConclusionsThe data revealed heterogeneity and highly dynamic distribution of NoV GI, GII, GIV, and SaV in environmental water in Chiang Mai, Thailand, during the study period of 2016–2018.

Molecular Epidemiology of GI.3 Norovirus Outbreaks from Acute Gastroenteritis Surveillance System in Taiwan, 2015–2019

Norovirus is the leading cause of food-borne disease outbreaks. We conducted this study to examine the incidence and molecular characteristics of norovirus genogroup I infections from acute gastroenteritis outbreaks in Taiwan. Between January 2015 and June 2019, 2121 acute gastroenteritis clusters were reported to Taiwan CDC, of which 351 (16.5%) clusters were positive for NoV GI, and GI.3 was the most prevalent (36.8%) during the study period. The GI.3 infections were significantly higher than non-GI.3 infections in the age groups of 0–5 and 6–18 years. The phylogenetic analysis of the MCC tree revealed that VP1 genes were divided into 3 groups: the GI.P3-GI.3 strains in Taiwan were genetically close to Japan and the GI.Pd-GI.3 strains were segregated into 2 other groups which were genetically closely related to China. In addition, 7 GI.Pd-GI.3 recombinants were identified circulating in Taiwan between 2018 and 2019, and the prevalence of GI.Pd-GI.3 should be monitored to assess whether this could become the new predominant strains in neighboring Asian countries or other parts of the world. Both GI.P3-GI.3 and GI.Pd-GI.3 strains cocirculate, the recombination among these two lineages occurs frequently, contributing to the genetic diversity and multiple occurrences of different norovirus lineages, and their rapid evolution makes future control more difficult. Continued surveillance and timely interventions are critical to understand the complexity of norovirus gene variation and to monitor the new emerging norovirus strains.

Etiology and epidemiological characteristics of gastroenteritis virus in food-borne diarrhea from three cities in Shandong Province, 2017

Objective: To analyze the etiology and epidemiological characteristics of gastroenteritis virus in foodborne diseases from three cities in Shandong. Methods: From January to December 2017, six sentinel hospitals in Jinan, Yantai and Linyi city of Shandong Province were selected as the research sites. Stool samples of 1 397 diarrhea patients were collected, as well as basic information and clinical symptoms. Duplex quantitative RT-PCR was used to detect Norovirus genogroupⅠ (Nov GⅠ) and genogroupⅡ (Nov GⅡ), Sapovirus (SAV) and Human astrovirus (HAstV), respectively, quantitative RT-PCR was used to detect group A Rotavirus (RVA), and quantitative PCR was used to detect Enteric adenovirus (EAdV). The specific gene of the virus were sequenced and typed. It was compared that the gastroenteritis virus rate in cases with different characteristics and the clinical symptoms difference between the virus positive and negative cases. Results: The median age (P(25), P(75)) was 23 (1, 42) , mainly male, 57.48% with 803 cased and children under 5 years old, 36.36% with 508 cases. The positive rate of gastroenteritis virus was 33.93% (474 cases), and that of Jinan, Linyi and Yantai City were 32.03% (147/459), 41.54% (189/455) and 28.57% (138/483), respectively (P<0.001). Nov GⅡ had the highest positive rate, 16.54% (231 cases), which, mainly GⅡ.P16/GⅡ.2 (48.28%, 56/116), peaked in May (24.75%, 50/202) and June (19.59%, 38/194). In patients of gastroenteritis virus positive, 44.51% (211/474) had vomiting symptoms, higher than that of patients of gastroenteritis virus negative (34.13%, 315/923). The difference was statistically significant (P<0.001). Conclusion: In Shandong Province, the majority of gastroenteritis patients were male and children under 5 years old. Nov GⅡ possessed highest epidemic intensity, and peaked in spring and summer. Viral gastroenteritis had atypical clinical symptoms.

Evaluation of Norovirus contamination in bivalve molluscs harvested from Northern Adriatic Sea, Italy

Abstract Background Norovirus (NoV) is the leading cause of acute gastroenteritis in developed countries. Transmission is through direct contact, unsanitary food handling or ingestion of contaminated water or food. Shellfish bioaccumulate NoV and current post-harvest depuration is not effective for its removal. Materials and Methods A monitoring programme for NoV in bivalve shellfish harvested from Northern Adriatic Sea was initiated in 2016. From January 2016 to March 2019, 418 samples including 257 Manila clams, 73 mussels, 46 striped clams, and 42 oysters, were examined. Mollusc samples were tested for NoV genogroups I (NoV GI) and II (NoV GII) contamination by RealTime RT-PCR according to ISO 15216-2. Results Ninety-three out of 418 tested samples (22.2%) were contaminated by at least one NoV genogroup, the simultaneous presence of the two genogroups was detected in 19/418 of the cases. Positive samples were distributed among the tested species as follow: 29/73 (39.7%) mussels, 51/257 (19.5%) Manila clams, 8/42 (19.0%) oysters, 5/46 (10.9%) striped clam. In 2016 (n = 135), 2017 (n = 122), 2018 (n = 132) and in the first trimester of 2019 (n = 12), prevalence of NoV was 6.7%, 24.6%, 31.8% and 41.4%, respectively. NoV GII was largely predominant being detected, alone or in association with GI, in 98.9% of the contaminated samples. On the other hand, GI prevalence increased from 1.5% (2016) to 12.1% in 2018, maybe reflecting a higher circulation of this genogroup in production environments. The vast majority of positive samples (91.4%) were detected during the cold season (November to March). Conclusions Overall positive samples have increased over the years (from 6.7% in 2016 to 31.8% in 2018). NoV GII was the most frequently detected genogroup, but NoV GI prevalence significantly raised in 2018. The routine application of quantitative RT-PCR (ISO 15216-1) to determine the viral load in bivalve molluscs would expand knowledge on potential for foodborne transmission. Key messages Overall positive samples have increased over the years. The routine application of quantitative PCR to determine the viral load in bivalve molluscs would expand knowledge on potential for foodborne transmission.

Molecular detection of enteric viruses in the stool samples of children without diarrhea in Bangladesh

A number of molecular epidemiological studies reported the detection of enteric viruses in asymptomatic children. The role of these viruses in an asymptomatic infection remains unclear. This study investigated the enteric viruses in the stool samples collected from children without diarrhea. Stool samples were collected during June to October 2016, from 227 children who lived in Matlab, Bangladesh. Seventeen enteric viruses, including rotavirus A, B, and C (RVA, RVB, and RVC), norovirus GI (NoV GI), norovirus GII (NoV GII), sapovirus (SaV), adenovirus (AdV), human astrovirus (HAstV), Aichivirus (AiV), human parechovirus (HPeV), enterovirus (EV), human bocavirus (HBoV), Saffold virus (SAFV), human cosavirus (HCoSV), bufavirus (BufV), salivirus (SalV), and rosavirus (RoV), were investigated by RT-PCR method. One hundred and eighty-two (80.2%; 182/227) samples were positive for some of these viruses, and 19.8% (45/227) were negative. Among the positive samples, 46.7% (85/182) were a single infection, and 53.3% (97/182) were coinfection with multiple viruses. The HCoSV was the most prevalent virus (41.4%), followed by EV (32.2%), NoV GII (25.6%), HPeV (8.8%), RVA (6.2%), AdV (5.7%), AiV (5.3%), SAFV (4.4%), and SaV (2.6%). Each of NoV GI, HAstV, HBoV, and BufV was detected at 0.4%. However, RVB, RVC, SalV, and RoV were not detected in this study. Phylogenetic analysis showed that diverse HCoSV species and genotypes were circulating in Bangladesh, and four strains of species A are proposed to be new genotypes. The data indicated that non-diarrheal Bangladeshi children were asymptomatically infected with wide varieties of enteric viruses.

Occurrence of pathogens in the river–groundwater interface in a losing river stretch (Besòs River Delta, Spain)

The aim of this study is to investigate the occurrence of faecal indicator and microbial pathogens (bacteria and virus) in the shallow urban aquifer of the Besòs River Delta (NE Spain). To this end, human adenovirus (HAdV) and Norovirus of genogroups I and II (NoV GI and NoV GII) as well as the faecal indicator bacteria (FIB) Escherichia coli (EC) and faecal enterococci (FE) were monitored in groundwater and in the River Besòs in December 2013 and in July 2104. None of the targeted pathogens were detected in groundwater in December 2013 but contamination of human origin was observed in approximately 50% of the points sampled in July 2014 reaching concentrations up to 99 GC/100 mL for HAdV. Generally, microbial concentrations in river water were higher than those detected in groundwater. This observation indicates that pathogens are naturally attenuated when river water infiltrates and flows through the aquifer, however HAdV were detected at a sampling point located at 380 m from the river in the absence of FIB. The presence of human viral contamination may represent a risk for the use of groundwater as a drinking water source. Further research is needed to understand the dynamics of pathogens in river–groundwater interface over long time periods and a wide range of flow conditions (wet and dry periods) since the urban groundwater of this aquifer might be a valuable drinking water resource in Barcelona especially during drought periods. The methodology followed in this research can be applied to other urban aquifers with similar purposes since the scarcity and contamination of freshwater resources are worldwide issues.

Occurrence and molecular characterization of enteric viruses in bivalve shellfish marketed in Vietnam

Viral contamination of bivalve molluscs is an extensively document food safety issue. Limited data are available, however, on the prevalence and quantitative levels of such viruses in countries, as Vietnam, with an important shellfish production, both for internal consumption and for export. The aim of this study was to evaluate both the occurrence and the quantitative levels of different viruses, including Norovirus (NoV genogroups I, II and IV), Hepatitis A (HAV), Hepatitis E (HEV), Astrovirus (AstV), and Aichi virus (AiV), in bivalve shellfish commercialized in Vietnam.A total of 121 samples (63 Pacific oysters and 58 white hard clams) were collected over nine months in two fish markets and two supermarkets in Hanoi, Vietnam. Norovirus was the most frequently detected virus (81.8% of samples) with viral loads ranging from below the quantification limit (LOQ) to 2.8 × 104 genome copies (g.c.)/g of shellfish digestive tissue and an average contamination of 3.8 × 103 g.c./g. HAV, HEV and AstV were detected in 1.7%, 11.6% and 12.4% of samples respectively (average concentrations: 1.3 × 102 g.c./g for HAV, 1.2 × 102 for HEV and 1.5 × 103 for AstV). AiV were also frequently detected (11.6% of samples), while NoV GIV was never observed. Contamination by at least one virus and by more than one virus was present in 83.5% and 52.9% of samples, respectively. Molecular characterization of NoV revealed circulation, over the observed period, of a wide variety of strains (four and nine genotypes for NoV GI and NoV GII respectively: GI.2, GI.4, GI.5, GI.6, GII.3, GII.4, GII.6, GII.7, GII.13, GII.14, GII.17, GII.18, GII.21).To our knowledge, this study is the first to provide quantitative information on the occurrence of enteric viruses in bivalve shellfish marketed in Vietnam, which will help outlining monitoring plans and setting appropriate control measures.

Quantitative PCR-based identification of enteric viruses contaminating fresh produce and surface water used for irrigation in Egypt.

Fresh produce irrigated with surface water that may contain pathogens such as enteric viruses can lead to outbreaks of foodborne viral illnesses. In the current study, we performed real-time PCR (qPCR) to monitor the presence of enteric viruses such as human adenoviruses (HAdVs), hepatitis A virus (HAV), rotavirus group A (RVA), and norovirus GI (NoV GI) in surface water and fresh produce that were grown using this surface water in Egypt. Samples were collected on four occasions from different sites located in the Delta and in Greater Cairo, Egypt. Of the 32 water samples and 128 fresh produce samples, 27/32 (84.3%) and 99/128 (77.3%), respectively, were positive for at least one virus. HAdV (30/32) with a mean viral load = 1.5 × 107 genome copies/L (GC/L) was the most commonly detected virus in water, followed by RVA (16/32, with a mean viral load = 2.7 × 105 GC/L), HAV (11/32, with a mean viral load = 1.2 × 104 GC /L), and NoV GI (10/32, with a mean viral load = 3.5 × 103 GC/L). Additionally, HAdV (71/128, with a mean viral load = 9.8 × 105 GC/g) was also the most commonly detected virus in the fresh produce, followed by NoV GI (43/128, with a mean viral load = 4.5 × 103 GC/g), HAV (33/128, with a mean viral load = 6.4 × 103 GC/g), and RVA (25/128, with a mean viral load = 1.5 × 104 GC/g). Our results indicate that fresh produce may be contaminated with a wide range of enteric viruses, and these viruses may originate from virus-contaminated irrigation water. Moreover, this fresh produce may serve as a potential vector for the transmission of viral foodborne illnesses. These findings are important for future risk assessment analysis related to water/foodborne viruses. Graphical abstract . Please provide caption for Graphical AbstractGraphical abstract showing sample collection and processing.

Prevalence of Norovirus in produce sold at retail in the United Kingdom

To acquire data on contamination with Norovirus in berry fruit and salad vegetables in the United Kingdom, one thousand one hundred and fifty two samples of fresh produce sold at retail in the UK were analysed for Norovirus. Of 568 samples of lettuce, 30 (5.3%) were Norovirus-positive. Most (24/30) lettuce samples which tested positive for Norovirus were grown in the UK and 19 of those 24 samples contained NoV GI. Seven/310 (2.3%) samples of fresh raspberries were Norovirus-positive. Most (6/7) of the positively-testing fresh raspberry samples were imported, but no predominance of a genogroup, or any seasonality, was observed. Ten/274 (3.6%) samples of frozen raspberries were Norovirus-positive. The country of origin of the positively-testing frozen raspberry samples was not identified in most (7/10) instances. The collected data add to the currently limited body of prevalence information on Norovirus in fresh produce, and indicate the need for implementation of effective food safety management of foodborne viruses.

Assessment of viral and bacterial contamination of fresh and ripened semi-hard cheeses.

This study aimed to evaluate viral and bacterial contamination from typical Brazilian cheeses, such as Minas (fresh) and Prato (ripened), commercially obtained in the Greater Metropolitan Region of the State of Rio de Janeiro, Brazil. Minas [30], Prato [30] and sliced Prato [30] cheese samples were investigated for norovirus genogroup I and II (NoV GI-II) and human adenovirus (HAdV) by direct nucleic acid extraction using TRIzol and amplification by TaqMan based quantitative polymerase chain reaction. Listeria monocytogenes, Salmonella spp., coagulase-positive staphylococci (CPS) and fecal coliforms were also assessed by using standard counting methods. NoV GI and GII were detected in one sample (1.1%) each and HAdV in nine samples (10.0%) while bacteriological analysis revealed five samples (5.5%) contaminated with L. monocytogenes, 27 (30.0%) with fecal coliforms and 10 (11.1%) with CPS. Salmonella spp. was not detected in any sample. Viruses were detected in 11 samples (12.2%), of which 9 met the microbiological criteria used to evaluate the microbiological quality of the cheeses, stressing the importance of considering virological parameters for monitoring this food matrix.

Irrigating Lettuce with Wastewater Effluent: Does Disinfection with Chlorine Dioxide Inactivate Viruses?

Reclaimed water obtained from urban wastewater is currently being used as irrigation water in water-scarce regions in Spain. However, wastewater can contain enteric viruses that water reclamation treatment cannot remove or inactivate completely. In the present study, greenhouse-grown baby lettuce ( L.) was irrigated with secondary treatment effluent from a wastewater treatment plant untreated and treated using chlorine dioxide (ClO). The effect of ClO treatment on the physicochemical characteristics and the presence of enteric viruses in irrigation water and lettuce was assessed. The presence of human noroviruses genogroups I and II (NoV GI and NoV GII), and human astroviruses (HAstV), was analyzed by real-time polymerase chain reaction (RT-qPCR). Additionally, to check for the loss of infectivity induced by the disinfection treatment, positive samples were re-analyzed after pretreatment with the intercalating dye PMAxx before RNA extraction and RT-qPCR. There were no significant differences in the proportion of positive samples and the concentration of enteric viruses between treated and untreated reclaimed water without PMAxx pretreatment ( > 0.05). A significantly lower concentration of NoV GI was detected in ClO-treated water when samples were pretreated with PMAxx ( < 0.05), indicating that inactivation was due to the disinfection treatment. Laboratory-scale validation tests indicated the suitability of PMAxx-RT-qPCR for discrimination between potentially infectious and ClO-damaged viruses. Although the applied ClO treatment was not able to significantly reduce the enteric virus load of the secondary effluent from the wastewater treatment plant, none of the lettuce samples analyzed ( = 36) was positive for the presence of NoV or HAstV.

Comprehensive Study on Enteric Viruses and Indicators in Surface Water in Kyoto, Japan, During 2014-2015 Season.

Certain enteric viruses that are present in the water environment are potential risk factors of waterborne infections. To better understand the impact of viruses in water, both enteric viruses and their potential indicators should be comparatively investigated. In this study, occurrences of GI- and GII-noroviruses (NoVs), sapovirus (SaV), rotavirus (RoV), Aichi virus 1 (AiV-1), enterovirus (EV), and pepper mild mottle virus (PMMoV) were quantitatively determined in surface water samples in Japan. Additionally, the genotype distribution of GI- and GII-NoVs was determined using a next-generation amplicon sequencing. PMMoV was the most abundant virus regardless of season and location, indicating its usefulness as an indicator for the viral contamination of water. Other potential indicators, AiV and EV, were less abundant than GII-NoV. Viruses other than PMMoV showed seasonality, i.e., EV and other viruses (NoVs, SaV, RoV, and AiV-1) became prevalent during summer and winter, respectively. SaV showed a relatively high abundance at a location that was affected by untreated wastewater. Regarding NoV genotypes, GI.1, GI.2, GI.4, GI.5, GI.6, GII.3, GII.4, GII.6, and GII.17 were found from the surface water samples. GII.4 and GII.17 seemed to have contributed to the high abundance of GII-NoV in the samples. Interestingly, GII.17 strains became prevalent in the water samples before becoming prevalent among gastroenteritis patients in Japan. These findings provide further insights into the properties of viruses as contaminants in the water environment.

Noroviruses in raw sewage, secondary effluents and reclaimed water produced by sand-anthracite filters and membrane bioreactor/reverse osmosis system

The importance of noroviruses (NoVs) in the epidemiology of waterborne diseases has increased globally in the last decades. The present study aimed to monitor genogroup I and II noroviruses in different treatment stages of four wastewater treatment plants (WWTPs) in the metropolitan São Paulo. WWTPs consist of secondary (activated sludge) and tertiary treatments (coagulation, sand-anthracite filters, membrane bioreactor (MBR)/reverse osmosis (RO) and chlorination). Raw sewage (500mL) and treated effluents (1L) were concentrated by celite and reclaimed water (40L) by hollow-fiber ultrafiltration system. Quantitative (qPCR) and nested PCR with nucleotide sequencing were used for quantification and molecular characterization. NoVs were widely distributed in raw wastewater samples (83.3%–100% NoV GI and 91.6%–100% NoV GII) and viral loads varied from 3.8 to 6.66log10gcL−1 for NoV GI and 3.8 to 7.3log10gcL−1 for NoV GII. Mean virus removal efficiencies obtained for activated sludge processes ranged from 0.3 to 0.8 log10 for NoV GI and 0.4 to 1.4 log10 for NoV GII. NoVs were not detected in the reuse water produced by MBR/RO system, while sand-anthracite filters resulted in a NoV GI and GII decay of 1.1–1.6 log10 and 0.7–1.6 log10, respectively. A variety of genotypes (GI.2, GI.3a, GI.3b, GI.5, GII.1, GII.4 Sydney 2012, GII.5, GII.6, GII.17) was observed, with a predominance of GI.2 and GII.17 in the different genogroups. These results corroborate with recent data about the entry and dissemination of the emerging genotype GII.P17-GII.17 Kawasaki 2014 in the country, and may indicate a change in the epidemiological patterns of norovirus strains circulation in this region. This is the first large-scale study to evaluate burden and genotypes of noroviruses in WWTPs in Brazil, providing a rapid diagnosis of viruses circulating in the population.

Occurrence of selected viral, bacterial and protozoan pathogens in fresh juices and smoothies in Prague, Czech Republic

Fresh squeezed juices and smoothies currently enjoy great popularity among consumers. However, data regarding the microbiological safety of these products sold to the public in food outlets are either very limited or missing completely. Within the current study, 80 samples of fresh juices and smoothies collected from food outlets in Prague, Czech Republic, were examined using culture and molecular methods for the presence of selected viral, bacterial and protozoan agents. One fifth of the smoothies and juices collected at food outlets contained Staphylococcus aureus, Yersinia enterocolitica, Cryptosporidium sp. or Giardia lamblia. The number of pathogens in all samples was lower than 50 CFU/g or 10 genome equivalents/g; in most cases pre-enrichment was required for successful culture of bacteria. Mycobacteria, human noroviruses (NoV GI and NoV GII) and adenoviruses were not detected at all. Smoothies and fresh juices are consumed without any further (thermal) treatment. Thus, the presence of pathogenic agents could have a significant impact on consumer health.

MW polyomavirus in diarrheal Italian infants.

MXPyV, like MWPyV, was identified in stool samples from children suffering diarrhea in Mexico. In this study, we used a home-made real time PCR to investigate the presence of this novel viruses in stool specimen collected from under-Five- Year-Old Children with gastroenteritis. A total of 192 fecal specimens previously screened for RV, ADV, NoV, HPeV and SaV, were tested for MWPyV with Taqman real time PCR. The most commonly detected virus was NoV GII (33.8%), followed by RV (21.3%), SaV (10.9%), HPeV(8%), NoV GI (6.7%) and AdV (1%). Real time PCR detected MWPyV in 1/192 (0.5%) patients. We detected MWPyV in 0.5% of fecal specimens collected from pediatric patients suffering gastroenteritis which is smaller than the previously reported in literature (4.4% in Australia and 12% Mexico).

Emergence of GI.6 Outbreaks in a High School in Fangshan District, Beijing, China

[Background] An outbreak of gastroenteritis happened in a high school in Fangshan District, Beijing, China in March 2016，in which 44 students developed the symptoms of vomiting, nausea, diarrhea and abdominal pain. Epidemiological investigation, laboratory investigation and Statistical analysis were conducted to identify the agent and source of this outbreak.[Methods] A case–control study was performed to discover the factors associated with this infection. Questionnaires and clinical data from those infection-exposed people were assessed. Here an outbreak case was defined as a student who developed at least three following symptoms: diarrhea, vomiting, abdominal pain and nausea in 72 hours after the lunch on 2nd March., and controls were those who study or work in the school but did not have lunch on the day. Samples (Stool samples, rectal swabs and vomit samples) from infection-exposed students and controls, as well as environmental samples like food were collected to test for the existence of any suspicious bacteria or viruses. Detection of norovirus wasdone by real-time TaqMan RT-PCR and sequence analysis. Chi-square test was used to decide whether those differences could be of any statistical significance. [Results] The total number of respondents with complete data was 44. An epidemiological data combined with the epidemic curve indicated that the outbreak started initially from a point source type, and was followed by a secondary transmission. Five stool samples, three rectal swabs and one vomit sample from case group turned out to be positive for norovirus genotype I by real-time PCR, and two asymptomatic food handlers in control group were positive for NoV GI. Sequence analysis of GI positive sample confirmed that the norovirus GI.6 variant was the etiological agent of the outbreak. Comparasion between the tested results from samples of those students who had lunch in school canteen and those who did notindicates that the difference of incidence was of statistical significance (P<0.01, 95% CI:4.22-324.41). Food and environmental samples were tested to be bacteria-negative. [Conclusions] Our result suggested that the etiological agent of the outbreak was norvirus GI.6, which leads to gastroenteritis. We identified that asymptomatic infected food handlers were most likely to be the source of the outbreak. Therefore，hand hygiene practices strict adherence to regulations and access to hand washing facilities should be strengthened.

Prevalence of Human Parainfluenza Viruses and Noroviruses Genomes on Office Fomites

The aim of this study was to evaluate the potential role of office fomites in respiratory (human parainfluenza virus 1—HPIV1, human parainfluenza virus 3—HPIV3) and enteric (norovirus GI—NoV GI, norovirus GII—NoV GII) viruses transmission by assessing the occurrence of these viruses on surfaces in office buildings. Between 2016 and 2017, a total of 130 surfaces from open-space and non-open-space rooms in office buildings located in one city were evaluated for HPIV1, HPIV3, NoV GI, and NoV GII viral RNA presence. Detection of viruses was performed by RT-qPCR method. Study revealed 27 positive samples, among them 59.3% were HPIV3-positive, 25.9% HPIV1-positive, and 14.8% NoV GII-positive. All tested surfaces were NoV GI-negative. Statistical analysis of obtained data showed that the surfaces of office equipment including computer keyboards and mice, telephones, and desktops were significantly more contaminated with respiratory viruses than the surfaces of building equipment elements such as door handles, light switches, or ventilation tracts (χ2p = 0.006; Fisher’s Exact p = 0.004). All examined surfaces were significantly more contaminated with HPIVs than NoVs (χ2p = 0.002; Fisher’s Exact p = 0.003). Office fomites in open-space rooms were more often contaminated with HPIVs than with NoVs (χ2p = 0.016; Fisher’s Exact p = 0.013). The highest average concentration of HPIVs RNA copies was observed on telephones (1.66 × 102 copies/100 cm2), while NoVs on the light switches (1.40 × 102 copies/100 cm2). However, the Kruskal–Wallis test did not show statistically significant differences in concentration levels of viral RNA copies on surfaces between the all tested samples. This study unequivocally showed that individuals in office environment may have contact with both respiratory and enteric viral particles present on frequently touched surfaces.

Changing pattern of prevalence, genetic diversity, and mixed infections of viruses associated with acute gastroenteritis in pediatric patients in New Delhi, India.

There are very few studies that have assessed multiple viral agents causing Acute-Gastroenteritis (AGE) in India. The present study compared the changing pattern of prevalence and genetic diversity of five enteric viruses associated with acute-diarrhea in Delhi children within a gap of 5 years. Fecal samples were collected from diarrheal children (<4 years) during two winter seasons: year 2009-2010 (n = 59) and year 2014-2015 (n = 85). Samples were individually tested for rotavirus-A, norovirus, astrovirus, adenovirus, and sapovirus using EIA/RT-PCR and genetically characterized by phylogenetic analysis. Rotavirus was the most predominant (54.9%) virus followed by norovirus (25.7%), astrovirus (8.3%), and adenovirus (4.9%) with rare detection of sapovirus (0.7%). While detection rate increased for both rotavirus (49.2-58.8%) and astrovirus (5.1-10.6%), norovirus detection rate decreased (30.5-22.4%) from 2009 to 2015. During the same time period, adenovirus detection remained low (4.7-5.1%). Interestingly, mixed infections increased from 8.5% to 16.5% after 5 years. G1P[8] rotavirus strain was found most predominant (40%). Both type-1 and 8 astroviruses were detected. Single sapovirus detected was of genotype GII.1. Both GI (GI.5, GI.3) and GII (GII.1, GII.4, GII.7, GII.21, GII.13) genogroups of norovirus were detected. Of particular significance was the first detection of other NoV genotypes (besides GII.4 and GI.3) in Delhi. This is also the first report of NoV GI.5 from India. A change in prevalence pattern and increased diversity from 2009 to 2015 emphasizes the need for continued enteric virus surveillance to help measure the impact of new diarrhea vaccine(s) introduced in India.

Outbreak of caliciviruses in the Singapore military, 2015

BackgroundFrom 31 August to 9 September 2015, a total of 150 military personnel at a military institution in Singapore were infected with acute gastroenteritis (AGE) with an attack rate of approximately 3%. This study aimed to determine the epidemiology of the outbreak, investigate its origins, and discuss measures to prevent future occurrences.MethodsAfter the AGE outbreak was declared on 31 August 2015, symptom surveys, hygiene inspections, and the testing of water, food, and stool samples were initiated. We collected 86 stool samples from AGE cases and 58 samples from food-handlers during the course of the outbreak and these stool samples were tested for 8 bacterial pathogens and 2 viral pathogens (i.e., norovirus and sapovirus).ResultsWe detected Sapovirus (SaV), group I Norovirus (NoV GI) and group II Norovirus (NoV GII) from the stool samples of AGE cases. Further sequence analyses showed that the AGE outbreak in August was caused mainly by three rarely reported calicivirus novel genotypes: NoV GI.7, NoV GII.17 and SaV GII.3. Control measures implemented focused on the escalation of personal and environmental hygiene, which included the separation of affected and unaffected soldiers, enforcement of rigorous hand-washing and hygiene, raising awareness of food and water safety, and disinfection of communal areas with bleach.ConclusionsThis study identified both NoV and SaV as the causative agents for an AGE outbreak at a Singapore military camp in August 2015. This study is also the first to report SaV as one of the main causative agents, highlighting the importance of caliciviruses as causative agents of AGE outbreaks in the Singapore military. As there are no commercially available vaccines against caliciviruses, strict personal hygiene and proper disinfection of environmental surfaces remain crucial to prevent calicivirus outbreak and transmission.