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There is a large burden of norovirus disease in child-care centers in Australia and around the world. Despite the ubiquity of norovirus outbreaks in child-care centers, little is known about the extent of this burden within the child-care center and the surrounding household clusters. Therefore, we performed an in-depth analysis of a gastroenteritis outbreak to examine the patterns of transmissions, household attack rates and the basic reproduction number (R0) for Norovirus in a child-care facility. We used data from parental interviews of suspected cases sent home with gastroenteritis at a child-care center between 24th of August and 18th of September 2020. A total of 52 persons in 19 household clusters were symptomatic in this outbreak investigation. Of all transmissions, 23 (46.9%) occurred in the child-care center, the rest occurring in households. We found a household attack rate of 36.5% (95% CI 27.3, 47.1%). Serial intervals were estimated as mean 2.5 ± SD1.45 days. The R0, using time-dependent methods during the growth phase of the outbreak (days 2 to 8) was 2.4 (95% CI 1.50, 3.50). The count of affected persons of a child-care center norovirus outbreak is approximately double the count of the total symptomatic staff and attending children. In the study setting, each symptomatic child-care attendee likely infected one other child-care attendee or staff and just over one household contact on average.

We investigated a large gastroenteritis outbreak that occurred in Northern Greece in 2019. A case was defined as anyone presenting with diarrhoea and/or vomiting from 24 January 2019 to 04 February 2019. We conducted a case-control study (CCS) using random selection of participants >16 years of age, residents of town X, who visited the health care centre between 25 and 28 January 2019. Moreover, we conducted a retrospective cohort study (CS) at the four elementary schools of the town. We collected clinical and water samples and the water supply system was inspected. In total, we recorded 638 cases (53% female; median age was 44 years (range 0–93)). Forty-eight cases and 52 controls participated in the CCS and 236 students in the CS. Both CCS and CS indicated tap water as the most likely source (OR 10, 95% CI 2.09–93.4, explaining 95.7% of cases; RR = 2.22, 95% CI 1.42–3.46, respectively). More than one pathogen was detected from stool samples of 6 of the 11 cases tested (norovirus, Campylobacter jejuni, Enterohemorrhagic E. coli (EHEC) and Enteropathogenic E. coli (EPEC)). Water samples, collected after ad-hoc chlorination, tested negative. Technical failures of the water tanks' status were identified. Our results suggested a waterborne outbreak. We recommended regular monitoring of the water supply system and immediate repair of technical failures.

Tese de Doutoramento em Ciências Veterinárias na especialidade de Segurança Alimentar Gastrointestinal infections have consistently been among the most frequent diseases and non-battle injuries, degrading operational effectiveness and force readiness in the military. Noroviruses appear as the most frequent agents causing acute gastrointestinal illness both in maritime and land theatres of operations, as well as in stationed troops or military training centers in many western countries. Nevertheless, little is known on the burden of norovirus gastroenteritis in the Portuguese military. In this work we set out to establish a gastroenteritis outbreak surveillance system (GOSS) to report disease cases to the Bromatology and Biologic Defense Laboratory during the five-year period, 2013-2017. During this period 14 gastroenteritis outbreaks were registered that affected a total of 410 military. Noroviruses were responsible for the majority of the outbreaks (8/14) and disease cases (210/410). Norovirus outbreaks occurred either in military exercises or in military bases and showed to have an impact on force readiness and operational effectiveness. In most cases the origin of the outbreaks was difficult to confirm but five (63%) were likely foodborne (including food handlers) or waterborne and three (37%) had predominant person-to-person transmission. Outbreaks were caused by both genogroup (G) I and II, but G II clearly outnumber those caused by G I (6/8) and case numbers (145/210). Norovirus GI was associated with waterborne outbreaks while GII was associated to foodborne and person-to-person transmission. Each outbreak was caused by a different genotype highlighting the high genetic diversity of the circulating noroviruses. Three of the identified genotypes (GI.9, GII.17 and GII.16-GII.2) were reported for the first time in Portugal. In conclusion, the present thesis identified norovirus as the most important etiologic agent of acute gastroenteritis outbreaks and the most frequent cause of food- and waterborne illness in the Portuguese Army. This work had contributed to change preventive measures and allowed the reinforcement of control actions that minimized the impact of norovirus outbreaks. RESUMO - Surtos de gastrenterite por norovírus no Exército Português - As infeções gastrointestinais encontram-se entre as doenças mais frequentemente reportadas como causa de diminuição da capacidade operacional e da prontidão de uma força militar. Os norovírus são os agentes mais vezes implicados na doença aguda gastrointestinal em teatros de operações, marítimos e terrestres, e em forças estacionadas ou centros de treino militares das Forças Armadas de Países Ocidentais. Esta tese teve como principal objetivo avaliar a relevância da gastrenterite aguda causada por norovírus no Exército Português. Foi estabelecido um sistema de vigilância no Exército, de modo que todos os surtos de gastroenterite são reportados ao Laboratório de Bromatologia e Defesa Biológica. Assim, verificou-se que entre 2013 e 2017 foram registados 14 episódios, que afetaram cerca de 410 militares, tendo os norovírus sido responsáveis pela maioria dos surtos (8/14) e dos casos (210/410). Os surtos ocorreram em exercícios militares e em bases do Exército com impacto na capacidade operacional e funcionamento regular das Unidades. Embora a origem destes tenha sido difícil de confirmar, acredita-se que 5 (63%) tiveram origem alimentar (incluindo os manipuladores de alimentos e a água) e que em 3 (37%) predominou a transmissão pessoa a pessoa. Os surtos foram causados por norovírus dos genogrupos (G) I e II, contudo o G II foi responsável pelo maior número de surtos (6/8) e casos (145/210). O genogrupo I esteve associado à água e o genogrupo II à transmissão pessoa a pessoa ou aos alimentos. Cada surto foi causado por um genótipo distinto, refletindo a grande diversidade genética dos norovírus que se encontram em circulação. Três dos genótipos identificados (GI.9, GII.17 e GII.16-GII.2) foram reportados pela primeira vez em Portugal. Ao identificar o norovírus como o mais importante agente etiológico de surtos de gastroenterite aguda e de doença com origem nos alimentos e na água no Exército Português, o presente trabalho contribuiu para a alteração de medidas preventivas de doenças de origem alimentar e para o reforço de ações de controlo dos surtos causados por este agente. The Present work was partially supported by the Portuguese Army. The author wants to acknowledge the Portuguese Army for the conditions provided for the experimental work, the development of the thesis and the financial support. N/A

Vibrio cholerae is a human pathogen, which is transmitted by the consumption of contaminated food or water. V. cholerae strains belonging to the serogroups O1 and O139 can cause cholera outbreaks and epidemics, a severe life-threatening diarrheal disease. In contrast, serogroups other than O1 and O139, denominated as non-O1/non-O139, have been mainly associated with sporadic cases of moderate or mild diarrhea, bacteremia and wound infections. Here we investigated the virulence determinants and phylogenetic origin of a non-O1/non-O139 V. cholerae strain that caused a gastroenteritis outbreak in Santiago, Chile, 2018. We found that this outbreak strain lacks the classical virulence genes harboured by O1 and O139 strains, including the cholera toxin (CT) and the toxin-coregulated pilus (TCP). However, this strain carries genomic islands (GIs) encoding Type III and Type VI secretion systems (T3SS/T6SS) and antibiotic resistance genes. Moreover, we found these GIs are wide distributed among several lineages of non-O1/non-O139 strains. Our results suggest that the acquisition of these GIs may enhance the virulence of non-O1/non-O139 strains that lack the CT and TCP-encoding genes. Our results highlight the pathogenic potential of these V. cholerae strains.DATA SUMMARYSequence data were submitted to GenBank under the accession number SRLP00000000. The authors confirm that all supporting data and protocols have been provided within the article or through supplementary data files.Data statementAll supporting data, code and protocols have been provided within the article or through supplementary data files. Four supplementary tables are available with the online version of this article.

In May 2016 a Norovirus (NoV) gastroenteritis outbreak involved a high school class visiting a seaside resort near Taormina (Mascali, Sicily). Twenty-four students and a teacher were affected and 17 of them showed symptoms on the second day of the journey, while the others got ill within the following 2 days. Symptoms included vomiting, diarrhoea and fever, and 12 students required hospitalisation. Stool samples tested positive for NoV genome by Real-Time polymerase chain reaction assay in all 25 symptomatic subjects. The GII.P2/GII.2 NoV genotype was linked to the outbreak by ORF1/ORF2 sequence analysis. The epidemiological features of the outbreak were consistent with food/waterborne followed by person-to-person and/or vomit transmission. Food consumed at a shared lunch on the first day of the trip was associated to illness and drinking un-bottled tap water was also considered as a risk factor. The analysis of water samples revealed the presence of bacterial indicators of faecal contamination in the water used in the resort as well as in other areas of the municipal water network, linking the NoV gastroenteritis outbreak to tap water pollution from sewage leakage. From a single water sample, an amplicon whose sequence corresponded to the capsid genotype recovered from patients could be obtained.

In April 2016, an outbreak of gastrointestinal illness (4,136 cases) occurred in Catalonia, Spain. We detected high levels of norovirus genogroups I and II in office water coolers associated with the outbreak. Infectious viral titer estimates were 33-49 genome copies/L for genogroup I and 327-660 genome copies/L for genogroup II.

Ulam is a group of traditional Malay vegetables, which is typically consumed raw and recognized as a popular dish in Malay culture. A number of previous studies have highlighted that Ulam are commonly used by the Malaysian natives in treating stomach pain. However, the findings have not yet been scientifically proven. The aim of this study is to investigate the antibacterial activity of four types of Ulam, Anacardium occidentale, Solanum tarvum, Diplazium esculentum, and Euodia redlevi against three gastroenteric pathogens isolated from patients involved in gastroenteritis outbreak which were obtained from the National Public Health Laboratory, Sungai Buloh. Three methods were applied in this research. The first one was the phytochemical screening to test the presence of alkaloids, flavonoids, saponins, and phenols in the methanolic extracts of the four plants. The second method was determining the total phenolic content (TPC) using the Follin-Ciocalteu method and thirdly, the Kirby-Bauer Disk Diffusion method, to test the antimicrobial activity of all plant extracts. Phytochemical screening reveals the presence of all constituents in the four extracts. The Ulam extracts also have shown positive results in inhibiting the activity of three enteric pathogens, indicated by the presence of a zone of inhibition. The evidence from this study suggests that the four Malaysian salads (Ulam), which are commonly used by Malaysian natives as herbal remedies in treating stomach pain, does contain the bioactive compounds that are the potential to be natural antibacterial in treating gastrointestinal tract disease.

Raspberries have lately caused several human norovirus (HuNoV) outbreaks in Europe. In this study, we developed and evaluated for HuNoV reverse transcription (RT)-PCR detection in frozen raspberries extraction methods that have equal sensitivity but are less time-consuming than widely used methods based on polyethylene glycol (PEG) precipitation and chloroform-butanol purification. One method was applied to stored frozen raspberries linked to previous HuNoV outbreaks and berries on sale. In the virus elution-based Method 1, sparkling water eluted viruses most efficiently from the berries. Method 2, based on direct nucleic acid extraction with minor PEG supplement, yielded the highest number of positive findings (4 out of 9) at low virus concentration level of 100 genome copies HuNoV genogroup II per 25 g raspberries. Both methods showed approximately equal sensitivity to a method including PEG precipitation and chloroform-butanol purification. Two naturally contaminated berry samples linked to HuNoV outbreaks in 2006 and 2009 were still positive for HuNoV genogroup I, but all berry products purchased from a local store remained negative for HuNoV. In conclusion, this study presents two efficient and rapid methods which can be used in urgent HuNoV outbreak investigations, since the results of the virus analysis are available in a few hours.

We report here the complete genome sequence of a novel recombinant GII.P16-GII.1 norovirus identified from eight fecal samples collected during an acute gastroenteritis outbreak in Jinan, Shandong Province, China, in 2017. The virus had nucleotide identities of 99% and 91% in the ORF1 and ORF2 genes of related strains, respectively.

Norovirus (NoV) gastroenteritis outbreaks appear frequently in food service operations (FSOs), such as in restaurants and canteens. In this study the presence of NoV and adenovirus (AdV) genomes was investigated on the surfaces of premises, especially in kitchens, of 30 FSOs where foodborne gastroenteritis outbreaks were suspected. The objective was to establish a possible association between the presence of virus genomes on surfaces and a visual hygienic status of the FSOs. NoV genome was found in 11 and AdV genome in 8 out of 30 FSOs. In total, 291 swabs were taken, of which 8.9% contained NoV and 5.8% AdV genome. The presence of NoV genomes on the surfaces was not found to associate with lower hygiene level of the premises when based on visual inspection; most (7/9) of the FSOs with NoV contamination on surfaces and a completed evaluation form had a good hygiene level (the best category). Restaurants had a significantly lower proportion of NoV-positive swabs compared to other FSOs (canteens, cafeteria, schools etc.) taken together (p = 0.00014). The presence of a designated break room for the workers was found to be significantly more common in AdV-negative kitchens (p = 0.046). Our findings suggest that swabbing is necessary for revealing viral contamination of surfaces and emphasis of hygiene inspections should be on the food handling procedures, and the education of food workers on virus transmission. Electronic supplementary material The online version of this article (doi:10.1007/s12560-017-9291-7) contains supplementary material, which is available to authorized users.

In mid-2016, norovirus GII.2 emerged as a major cause of gastroenteritis outbreak in Japan with overall detection rate of 56.3% of norovirus cases. The differences in ORF1 and ORF2 of some norovirus GII were observed. Inter-subtype recombinants GII.Pe/GII.2, GII.P16/GII.2 and GII.P17/GII.2 were detected. Three amino acid substitutions were noted at P2 antigenic site of GII.P16/GII.2 recombinants. Furthermore, this study revealed that the current immunochromatographic kit available in Japan could be used effectively for the detection of recent GII.2 genotype.

An emerging recombinant norovirus GII.P16/GII.4 Sydney 2012 strain caused a gastroenteritis outbreak amongst attendees at a large health function in regional New South Wales, Australia. This was the third outbreak caused by the recombinant GII.P16/GII.4 Sydney 2012 strain in this region in 2017, which appears to be emerging as a common strain in the Hunter New England region.

Drinking water outbreaks occur worldwide and may be caused by several factors, including raw water contamination, treatment deficiencies, and distribution network failure. This study describes two drinking water outbreaks in Finland in 2016 (outbreak I) and 2018 (outbreak II). Both outbreaks caused approximately 450 illness cases and were due to drinking water pipe breakage and subsequent wastewater intrusion into the distribution system. In both outbreaks, the sapovirus was found in patient samples as the main causative agent. In addition, adenoviruses and Dientamoeba fragilis (outbreak I), and noroviruses, astroviruses, enterotoxigenic and enterohemorragic Escherichia coli (ETEC and EHEC, respectively) and Plesiomonas shigelloides (outbreak II) were detected in patient samples. Water samples were analyzed for the selected pathogens largely based on the results of patient samples. In addition, traditional fecal indicator bacteria and host-specific microbial source tracking (MST) markers (GenBac3 and HF183) were analyzed from water. In drinking water, sapovirus and enteropathogenic E. coli (EPEC) were found in outbreak II. The MST markers proved useful in the detection of contamination and to ensure the success of contaminant removal from the water distribution system. As mitigation actions, boil water advisory, alternative drinking water sources and chlorination were organized to restrict the outbreaks and to clean the contaminated distribution network. This study highlights the emerging role of sapoviruses as a waterborne pathogen and warrants the need for testing of multiple viruses during outbreak investigation.

During an investigation of an outbreak of gastroenteritis caused by Salmonella enterica serovar Paratyphi B variant L(+) tartrate(+), we identified unpasteurized tempeh as a novel food vehicle and Rhizopus spp. starter culture as the source of the contamination. Safe handling of uncooked, unpasteurized tempeh should be emphasized for prevention of foodborne illnesses.

Introduction This study aimed to monitor the seasonality of rotavirus infection, and gain insight into the variability of Brazilian strains. Methods A total of 28 stool samples were analyzed from 698 revised cases of gastroenteritis during a norovirus outbreak in the summer of 2010 in Guarujá, Brazil. Diagnosis was performed using enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and sequencing. Results Rotavirus infection was detected in 17.9% (5/28) of samples; 4 samples were G2P[4] genotype, and one G2P[4]+P[6] genotype. G2 and P[4] sequences showed a genetic relationship to strains from India and Russia, respectively. Conclusions The seasonal pattern of rotavirus may be a consequence of human activity apart from climate factors.

In March 2010, a massive outbreak of gastroenteritis started in the region of Antofagasta (northern Chile). The outbreak was mainly attributed to Norovirus genogroup II although ETEC strains were also isolated with high frequency from clinical samples. We review this outbreak and determined that ETEC was an underestimated etiologic agent.

The first human astrovirus was discovered in 1975 by Madeley and Cosgrove through visualization of viral particles in stool using electron microscopy. Over the course of the next ∼20 years, an additional seven serotypes of human astroviruses were discovered. In the last decade, new technological advances in microarrays and sequencing strategies enabled more sophisticated methods for detecting viruses in clinical and environmental specimens. These methods led to the discovery of many novel viruses from a variety of virus families. They also brought about a dramatic realization that more astroviruses can be found in humans than previously recognized. In fact, the number of astroviruses associated with humans has nearly doubled within the last few years. Furthermore, the discovery of novel astroviruses in human specimens revealed that there is more diversity amongst them than was assumed based on the high level of similarity between human astroviruses 1 and 8. This chapter will describe the discovery and early characterization of the five novel astroviruses initially identified in human stool samples in 2008 and 2009.

Hellenic Center for Disease Control and Prevention, Athens 15123, Greece; E-Mails: kmellou@gmail.com (K.M.); gspala@gmail.com (G.S.) * Author to whom correspondence should be addressed; E-Mail: avanta@upatras.gr; Tel.: +30-26-10969875; Fax: +30-26-10969875. Received: 20 July 2011; in revised form: 12 August 2011 / Accepted: 16 August 2011 / Published: 22 August 2011 Abstract: In June 2006, an outbreak alert regarding cases of acute gastroenteritis in a region in North Eastern Greece (population 100,882 inhabitants), triggered investigations to guide control measures. The outbreak started the first days of June, and peaked in July. A descriptive epidemiological study, a virological characterization of the viral agent identified from cases as well as a phylogenetic analysis was performed. From June 5 to September 3, 2006 (weeks 23–44), 1,640 cases of gastroenteritis (45.2% male and 54.8% female, aged 3 months to 89 years) were reported. The overall attack rate for the period was 16.3 cases/1,000 inhabitants. About 57% of cases observed were under the age of 15 years. Αnalysis of faecal samples identified Norovirus GII strains. Fifteen different Norovirus GII strains were recorded, presenting a homology of 94.8% (86–97%) to GII strains obtained from GenBank. The long duration of the outbreak suggests an important role of person-to-person transmission, while the emergence of the outbreak was possibly due to contaminated potable water, although no viruses were detected in any tested water samples. This outbreak underscores the need for a national surveillance system for acute non-bacterial gastroenteritis outbreaks.

Noroviruses and rotaviruses from a gastroenteritis outbreak affecting >300 people near Garda Lake (Northern Italy) in 2009 were investigated. Characterization of viruses from 40 patient stool samples and 5 environmental samples identified three distinct rotavirus and five norovirus genotypes; two of the latter were detected in both patient and environmental samples.

Objectives: The molecular and epidemiological features of community-based norovirus-associated sporadic gastroenteritis incidents (NASGIs) are poorly understood. This study examined these features and compared the findings with studies of community-based and institutional norovirus-associated gastroenteritis outbreaks (NAGOs). Methods: Fecal specimens from NASGIs and NAGOs that occurred in Victoria, Australia (2002–2007) were tested for norovirus by reverse transcription-polymerase chain reaction methodology. Norovirus genotype was determined by nucleotide sequence analysis. Results: 106 community-based NASGIs, 116 community-based NAGOs and 902 institutional NAGOs were identified. The mean age and gender ratio of individuals associated with community-based NASGIs and community-based NAGOs were similar but differed from that found for institutional NAGOs. Although GII.4 was the predominant genotype associated with all three incident types, the mix of genotypes was similar for community-based NASGIs and community-based NAGOs but that for institutional NAGOs was different. All three incident types had a similar seasonal periodicity due to the pronounced seasonal periodicity of GII.4 incidents. Conclusions: The molecular and epidemiological features of noroviruses associated with community-based NASGIs and community-based NAGOs are similar but are different from those found for institutional NAGOs.

Gastroenteritis affected many British military personnel during the war in Iraq. In the first month, 1,340 cases were seen; 73% of patients required hospital admission and 36% were hospital staff. In a survey of 500 hospital staff, 76% reported gastroenteritis, which was more likely in clinical workers. Investigations showed only caliciviruses.

Salmonella enterica serovar Weltevreden and Vibrio fluvialis were identified as etiological agents of a foodborne gastroenteritis outbreak after an Iftar feast in North Dumdum. Of the 278 cases admitted to the Infectious Diseases Hospital, Kolkata, 44 stool samples were tested for the enteric pathogens. Six were positive for Salmonella Weltevreden, 5 for Vibrio fluvialis, and 8 contained both of the pathogens. Consumption of mutton-ghogni might have been the likely vehicle of this outbreak. In the pulsed-field gel electrophoresis, Salmonella Weltevreden was identified as a single clone but the V. fluvialis strains were heterogeneous.

Although Escherichia albertii is an emerging intestinal pathogen, it has been associated only with sporadic human infections. In this study, we determined that a human gastroenteritis outbreak at a restaurant in Japan had E. albertii as the major causative agent.

In July 2001, an outbreak of gastroenteritis occurred in Helsinki among children and adults after bathing in an outdoor wading pool. The epidemiological survey revealed that at least 242 persons were affected. Microbiological testing of both patient stool samples and of the pool water revealed the presence of two different gastroenteritis viruses: a norovirus (NV) and an astrovirus. Amplicon sequencing of the NV samples showed nucleotide sequence identity between the virus from patients and the water. After changing the pool water and the sand at the bottom of the pool followed by shock chlorination, no virus could be detected in the water. However, NV was continuously detected in the water outlet well as much as 8 months after the incident. Here we show how molecular methods aided in tracing the source of the epidemic and in finding the causative pathogens both in patients and in the environment.

Three different studies are reported concerning the environmental pollution caused by viruses in Albania. The first study describes an outbreak of gastroenteritis in the capital city, involving 2,722 children attending the Paediatric Unit of Tirana Hospital. The age group with the highest morbidity was 0-5 years, with 89.5%; no fatalities were recorded during the outbreak. Rotavirus was detected in 26/28 faecal samples by RT-PCR, although astrovirus, adenovirus and calicivirus were also present. The second study describes an outbreak of hepatitis A virus involving the city of Lac. Two hundred cases were recorded, with the highest incidence in the age-group 5-9 years. Phylogenetic analysis of the VP1/2A region showed the presence of a unique sequence: genotype IA. Rotavirus was identified in drinking-water samples collected during the outbreak. The third study describes the prevalence of HAV and HEV in 202 sera randomly collected from 12 different cities in Albania. HAV showed a high incidence (66.2%), whereas none was positive for HEV. The genomic analysis of the VP1/2A junction revealed the presence of only one genotype (IA) with few point mutations and just two amino acid substitutions at codons 22 and 34. Additionally, two potential antigenic variants were detected, the first at position 46 of VP3 and the second at position 23 of VP1.

During November 2015, the North Carolina Division of Public Health was notified by the Pitt County Health Department (PCHD) that approximately 40 persons who attended a catered company Thanksgiving lunch the previous day were ill with diarrhea and abdominal pain. The North Carolina Division of Public Health and PCHD worked together to investigate the source of illness and implement control measures. Within hours of notification, investigators developed and distributed an online survey to all lunch attendees regarding symptoms and foods consumed and initiated a cohort study. A case of illness was defined as abdominal pain or diarrhea in a lunch attendee with illness onset

Introduction: This study describes the investigation of a gastroenteritis outbreak in a group of students, associated with a dinner reunion in February 2013 in Porto, Portugal. Methodology: An anonymous structured questionnaire was developed and sent to 34 students who attended the dinner reunion. Eighteen students completed the questionnaire and thirteen met the case definition (attack rate of 72%). Stools from two students were screened for norovirus by RT-PCR using primer pairs that target the highly conserved polymerase gene and the capsid gene. Results: Norovirus genotyping confirmed the variant Sydney 2012 as the probable cause of the outbreak. Conclusion: This is the first report of an outbreak associated with the new variant Sydney 2012 in Portugal. The study was supported by FEDER funds through Programa Operacional Factores de Competividade (COMPETE), by national funding through Fundação para a Ciência e a Tecnologia (FCT) (project PTDC/CVT/113218/2009), and by grant SFRH/BD/45407/2008, and by project Ovislab ICT-2013-05-004-5314 ID-64757.

Shellfish have been implicated in norovirus (NoV) infection outbreaks worldwide. This study presents data obtained from various batches of shellfish and fishery products from a Belgian seafood company over a 6-month period. For the intact shellfish (oysters, mussels, and clams), 21 of 65 samples from 12 of 34 batches were positive for NoVs; 9 samples contained quantitative NoV levels at 3,300 to 14,300 genomic copies per g. For the semiprocessed fishery products (scallops and common sole rolls with scallop fragments), 29 of 36 samples from all eight batches were positive for NoVs; 17 samples contained quantitative NoV levels at 200 to 1,800 copies per g. This convenience study demonstrated the performance and robustness of the reverse transcription quantitative PCR detection and interpretation method and the added value of NoV testing in the framework of periodic control of seafood products bought internationally and distributed by a Belgian seafood processing company to Belgian food markets.

In January of 2008, during the peak of the rotavirus season in Guatemala, a gastroenteritis outbreak with high mortality among infants was reported in Guatemala. Despite extensive efforts, the investigation was limited by the lack of bulk stool specimens collected, particularly from the more severely dehydrated or deceased children.We evaluated the diagnostic performance of rectal swab specimens compared with bulk stool for the detection of rotavirus and norovirus.Patients with diarrhea (≥3 loose stools in 24 h) were enrolled through an ongoing surveillance system in Guatemala. From January through March 2009, we attempted to enroll 100 patients5 years old captured by the diarrhea surveillance, and collected paired bulk stool and rectal swabs specimens from them. Specimens were tested for norovirus using real-time reverse transcription-polymerase chain reaction and for rotavirus via enzyme immunoassay.We enrolled 102 patients with paired specimens; 91% of 100 paired specimens tested for rotavirus yielded concordant results positive for rotavirus with a negativity rate of 83%. Among 100 paired specimens tested for norovirus, 86% were concordant norovirus detection and the negativity rate was 85%. The diagnostic performance for rotavirus and norovirus detection did not differ significantly between the two specimen types.Testing of properly collected fecal specimens using rectal swabs may be a viable alternative to bulk stool for detection of rotavirus and norovirus, particularly during outbreaks where collection of bulk stool may be difficult.

During September/October 2012, a norovirus gastroenteritis outbreak affecting about 11,000 people occurred in Germany. Epidemiological studies suggested that frozen strawberries represented the vehicle of infection. We describe here the analysis of frozen strawberries for the presence of norovirus. Samples were taken by applying a stratified subsampling scheme. Two different methods for virus extraction from strawberries were compared. First, viruses were eluted from strawberries under alkaline conditions and concentrated using a polyethylene glycol precipitation. Second, ultrafiltration was applied for concentration of viruses rinsed off of the berries. In both cases, RNA was extracted and analyzed by real-time RT-PCR. Application of the ultrafiltration method generally resulted in a lower detection rate. Noroviruses were detected in 7/11 samples derived from the lot of strawberries implicated in the outbreak using the precipitation method. Typing of norovirus revealed three different genotypes including a combination of norovirus genotype II.16 (viral polymerase) and II.13 (viral capsid). This genotype combination was also found in some of the patients that were involved in the outbreak, but that had not been reported in Germany so far. In conclusion, heterogeneously distributed noroviruses in frozen strawberries can be detected by applying an optimized combination of sampling procedures, virus extraction methods, and real-time RT-PCR protocols. The detection of several different genotypes in the strawberries may suggest contamination from sewage rather than from a single infected food handler.

Strains of Bacillus cereus and B. thuringiensis were tested by the Tecra VIA kit for the ability to produce a diarrhoeal enterotoxin. The strains of B. thuringiensis were isolated from commercial B. thuringiensis-based insecticides (BactimosTM, DiPelTM, FlorbacTM FC, ForayTM 48B, NovodorTM FC, TurexTM, VecTobacTM, XenTariTM). The production of diarrhoeal enterotoxin varied by a factor of more than 100 among the different strains tested. B. cereus (F4433/73) produced the highest amount of enterotoxin and the B. thuringiensis strain isolated from DiPelTM the lowest. The products were tested for their content of diarrhoeal enterotoxin and all products, except MVPTM which does not contain viable B. thuringiensis spores, contained diarrhoeal enterotoxins. The results indicates an potential risk for gastroenteritis outbreak caused by B. thuringiensis.

Copyright © 2012 Rajko-Nenow et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited., Norovirus (NoV) is the single most important agent of foodborne viral gastroenteritis worldwide. Bivalve shellfish, such as oysters, grown in areas contaminated with human faecal waste may become contaminated with human pathogens including NoV. A study was undertaken to investigate NoV contamination in oysters (Crassostrea gigas) from a shellfishery over a 24 month period from October 2007 to September 2009. Oyster samples were collected monthly from a commercial shellfish harvest area classified as category B under EU regulations, but that had had been closed for commercial harvesting due to its previous association with NoV outbreaks. Real-time reverse transcription quantitative PCR (RT-qPCR) was used to determine the concentration of human NoV genogroups I and II (GI and GII) in monthly samples. Total NoV (GI and GII) concentrations in NoV positive oysters ranged from 97 to 20,080 genome copies g− 1 of digestive tissue and displayed a strong seasonal trend with greater concentrations occurring during the winter months. While NoV GII concentrations detected in oysters during both years were similar, NoV GI concentrations were significantly greater in oysters during the winter of 2008/09 than during the winter of 2007/08. To examine the NoV genotypes present in oyster samples, sequence analysis of nested RT-PCR products was undertaken. Although NoV GII.4 is responsible for the vast majority of reports of outbreaks in the community, multiple NoV genotypes were identified in oysters during this study: GI.4, GI.3, GI.2, GII.4, GII.b, GII.2, GII.12, and GII.e. NoV GI.4 was the most frequently detected genotype throughout the study period and was detected in 88.9% of positive samples, this was followed by GII.4 (43.7%) and GII.b (37.5%). This data demonstrates the diversity of NoV genotypes that can be present in sewage contaminated shellfish and that a disproportionate number of non-NoV GII.4 genotypes can be found in environmental samples compared to the number of recorded human infections associated with non-NoV GII.4 genotypes.

Faecal specimens collected from two outbreaks of acute gastroenteritis that occurred in southern Mumbai, India in March and October, 2006 were tested for seven different enteric viruses. Among the 218 specimens tested, 95 (43.6%) were positive, 73 (76.8%) for a single virus and 22 (23.2%) for multiple viruses. Single viral infections in both, March and October showed predominance of enterovirus (EV, 33.3% and 40%) and rotavirus A (RVA, 33.3% and 25%). The other viruses detected in these months were norovirus (NoV, 12.1% and 10%), rotavirus B (RVB, 12.1% and 10%), enteric adenovirus (AdV, 6.1% and 7.5%), Aichivirus (AiV, 3% and 7.5%) and human astrovirus (HAstV, 3% and 0%). Mixed viral infections were largely represented by two viruses (84.6% and 88.9%), a small proportion showed presence of three (7.7% and 11%) and four (7.7% and 0%) viruses in the two outbreaks. Genotyping of the viruses revealed predominance of RVA G2P[4], RVB G2 (Indian Bangladeshi lineage), NoV GII.4, AdV-40, HAstV-8 and AiV B types. VP1/2A junction region based genotyping showed presence of 11 different serotypes of EVs. Although no virus was detected in the tested water samples, examination of both water and sewage pipelines in gastroenteritis affected localities indicated leakages and possibility of contamination of drinking water with sewage water. Coexistence of multiple enteric viruses during the two outbreaks of gastroenteritis emphasizes the need to expand such investigations to other parts of India.

A gastroenteritis outbreak affected 45 people (40 residents and five staff) in a nursing home for the elderly in the Celje region, north-east Slovenia, between 17 December and 31 December 2010. Rotavirus group A was laboratory confirmed in the stools of five ill individuals. The outbreak was identified when the number of affected persons was high but was successfully controlled after implementing preventive measures.

BACKGROUND: Norovirus GII.4 is the predominant genotype circulating worldwide over the last decade causing 80% of all norovirus outbreaks with new GII.4 variants reported in parallel with periodic epidemic waves of norovirus outbreaks. The circulating new GII.4 variants and the epidemiology of norovirus outbreaks in Alberta, Canada have not been described. Our hypothesis is that the periodic epidemic norovirus outbreak activity in Alberta was driven by new GII.4 variants evolving by genetic drift. METHODOLOGY/PRINCIPAL FINDINGS: The Alberta Provincial Public Health Laboratory performed norovirus testing using RT-PCR for suspected norovirus outbreaks in the province and the northern Territories between 2000 and 2008. At least one norovirus strain from 707 out of 1,057 (66.9%) confirmed norovirus outbreaks were successfully sequenced. Phylogenetic analysis was performed using BioNumerics and 617 (91.1%) outbreaks were characterized as caused by GII.4 with 598 assigned as novel variants including: GII.4-1996, GII.4-2002, GII.4-2004, GII.4-2006a, GII.4-2006b, GII.4-2008a and GII.4-2008b. Defining July to June of the following year as the yearly observation period, there was clear biannual pattern of low and high outbreak activity in Alberta. Within this biannual pattern, high outbreak activity followed the emergence of novel GII.4 variants. The two variants that emerged in 2006 had wider geographic distribution and resulted in higher outbreak activity compared to other variants. The outbreak settings were analyzed. Community-based group residence was the most common for both GII.4 variants and non-GII.4 variants. GII.4 variants were more commonly associated with outbreaks in acute care hospitals while outbreaks associated non-GII.4 variants were more commonly seen in school and community social events settings (p

Analysis of 316 outbreaks of gastroenteritis in France from September 2007 through March 2009 showed that genogroup II.4 (GGII.4) noroviruses were predominant and mostly belonged to the 2006b variant. However, the new GGII.4 variants, variant 2008 and the newly discovered Cairo variant from the Middle East, were also detected. The epidemiological survey suggests that these new variants might become the next predominant strains.

An atypically high level of norovirus activity was noticed in Hong Kong beginning in early May 2006. A study was carried out to investigate whether this was caused by a new norovirus variant. Epidemiological data including monthly positivity rates and the numbers of outbreaks per month from January to July 2006 were analyzed and compared to those from 2002 to 2005. In a comparison with the epidemiological data from 2001 to 2005, an atypical peak of norovirus-associated gastroenteritis outbreak was observed beginning in May 2006, concurring with a striking increase in norovirus activity. Most of the outbreaks (>60%) were located in homes for the elderly. Phylogenetic analysis for both RdRp and 5′ capsid regions showed that this epidemic was caused by a new genogroup II/4 variant. This variant was genetically distinct from the predominant variants of 2002 and 2004 but was closely related to one of the 95/96-subset variants which caused an epidemic in Hong Kong in 2001, suggesting that the 95/96 subset may be starting to recirculate.

[ES] Se presentan los resultados de los distintos estudios realizados en la investigación de un brote de gastroenteritis y se exponen las dificultades y las limitaciones que condicionaron su estudio. Se diseñaron varios estudios analíticos: un estudio de cohortes y un estudio de casos y controles con diferente tamaño muestral. De los 189 participantes, el 43% respondió el cuestionario. Se identificaron 45 casos. Los factores que se asociaron significativamente con la enfermedad fueron: la cena del día 11 de diciembre en uno de los restaurantes del complejo, tanto en el estudio de cohortes como en el de casos y controles, y la excursión a una isla cercana en el de cohortes. Se confirmó la existencia de un brote, aunque debido a la baja tasa de respuesta y las dificultades para entrevistar a los participantes del viaje, no fue posible identificar la vía de transmisión. [EN] We present the results of different studies performed in the investigation of a gastroenteritis outbreak and we describe the difficulties and limitations encountered during its study. Several analytical studies were designed: one cohort study and one case-control study with different samples sizes. Of the 189 trip participants, 43% answered the questionnaire. Forty-five cases were identified. The factors significantly associated with the disease were: dinner in one of the restaurants of the resort on December 11 in all analytical studies and the excursion to a nearby island in the cohort study. The outbreak was confirmed however due to the low response rate and the problems encountered to interview the participants in the trip, it was not possible to identify the route of transmission. Sí

To report an acute gastroenteritis outbreak caused by a genogroup 2 genotype 6 (GII.6) strain norovirus in Shanghai, China.Noroviruses are responsible for approximately half of all reported gastroenteritis outbreaks in many countries. Genogroup 2 genotype 4 strains are the most prevalent. Rare outbreaks caused by GII.6 strains have been reported. An acute gastroenteritis outbreak occurred in an elementary school in Shanghai in December of 2013. Field and molecular epidemiologic investigations were conducted.The outbreak was limited to one class in an elementary school located in southwest Shanghai. The age of the students ranged from 9 to 10 years. The first case emerged on December 10, 2013, and the last case emerged on December 14, 2013. The cases peaked on December 11, 2013, with 21 new cases. Of 45 students in the class, 32 were affected. The main symptom was gastroenteritis, and 15.6% (5/32) of the cases exhibited a fever. A field epidemiologic investigation showed the pathogen may have been transmitted to the elementary school from employees in a delicatessen via the first case student, who had eaten food from the delicatessen one day before the gastroenteritis episodes began. A molecular epidemiologic investigation identified the cause of the gastroenteritis as norovirus strain GII.6; the viral sequence of the student cases showed 100% homology with that of the shop employees. Genetic relatedness analyses showed that the new viral strain is closely related to previously reported GII.6 sequences, especially to a strain reported in Japan.This is the first report to show that norovirus strain GII.6 can cause a gastroenteritis outbreak. Thus, the prevalence of GII.6 noroviruses requires attention.

An international outbreak linked to oyster consumption involving a group of over 200 people in Italy and 127 total subjects in 13 smaller clusters in France was analyzed using epidemiological and clinical data and shellfish samples. Environmental information from the oyster-producing area, located in a lagoon in southern France, was collected to investigate the possible events leading to the contamination. Virologic analyses were conducted by reverse transcription-PCR (RT-PCR) using the same primer sets for both clinical and environmental samples. After sequencing, the data were analyzed through the database operated by the scientific network FoodBorne Viruses in Europe. The existence of an international collaboration between laboratories was critical to rapidly connect the data and to fully interpret the results, since it was not obvious that one food could be the link because of the diversity of the several norovirus strains involved in the different cases. It was also demonstrated that heavy rain was responsible for the accidental contamination of seafood, leading to a concentration of up to hundreds of genomic copies per oyster as detected by real-time RT-PCR.

An increase in gastroenteritis outbreaks due to Norovirus has been reported worldwide. We investigated a large-scale outbreak affecting 246 residents and 33 staff members in six nursing homes in the Tel-Aviv district, Israel, during 3 weeks in 2002. Person-to-person spread was noticed in all nursing homes. The spread of disease could not be attributed to social interactions. Among the elderly residents, the hospitalization rate was 10·2% and the case-fatality rate was 2·0%. Bacteriological cultures were negative. Overall, 7 out of 15 stool specimens were positive for Norovirus by RT–PCR. All were sequenced and found to be 90% identical. The characteristics of this outbreak and the RT–PCR results suggest that illness was caused by Norovirus. Due to the high case-fatality rate of Norovirus gastroenteritis, there should be a high index of suspicion when encountering a gastroenteritis outbreak among the elderly. This will enable prompt action to stop the spread of illness.