Molecular surveillance of norovirus, 2005-16: an epidemiological analysis of data collected from the NoroNet network.

BACKGROUND: The development of a vaccine for norovirus requires a detailed understanding of global genetic diversity of noroviruses. We analysed their epidemiology and diversity using surveillance data from the NoroNet network. METHODS: We included genetic sequences of norovirus specimens obtained from outbreak investigations and sporadic gastroenteritis cases between 2005 and 2016 in Europe, Asia, Oceania, and Africa. We genotyped norovirus sequences and analysed sequences that overlapped at open reading frame (ORF) 1 and ORF2. Additionally, we assessed the sampling date and country of origin of the first reported sequence to assess when and where novel drift variants originated. FINDINGS: We analysed 16 635 norovirus sequences submitted between Jan 1, 2005, to Nov 17, 2016, of which 1372 (8·2%) sequences belonged to genotype GI, 15 256 (91·7%) to GII, and seven (<0·1%) to GIV.1. During this period, 26 different norovirus capsid genotypes circulated and 22 different recombinant genomes were found. GII.4 drift variants emerged with 2-3-year periodicity up to 2012, but not afterwards. Instead, the GII.4 Sydney capsid seems to persist through recombination, with a novel recombinant of GII.P16-GII.4 Sydney 2012 variant detected in 2014 in Germany (n=1) and the Netherlands (n=1), and again in 2016 in Japan (n=2), China (n=8), and the Netherlands (n=3). The novel GII.P17-GII.17, first reported in Asia in 2014, has circulated widely in Europe in 2015-16 (GII.P17 made up a highly variable proportion of all sequences in each country [median 11·3%, range 4·2-53·9], as did GII.17 [median 6·3%, range 0-44·5]). GII.4 viruses were more common in outbreaks in health-care settings (2239 [37·2%] of 6022 entries) compared with other genotypes (101 [12·5%] of 809 entries for GI and 263 [13·5%] of 1941 entries for GII non-GII.Pe-GII.4 or GII.P4-GII.4). INTERPRETATION: Continuous changes in the global norovirus genetic diversity highlight the need for sustained global norovirus surveillance, including assessment of possible immune escape and evolution by recombination, to provide a full overview of norovirus epidemiology for future vaccine policy decisions. FUNDING: European Union's Horizon 2020 grant COMPARE, ZonMw TOP grant, the Virgo Consortium funded by the Dutch Government, and the Hungarian Scientific Research Fund.

Shigellosis in refugees, Austria, July to November 2015.

We report on a cluster of shigellosis including 21 cases in refugees and two in local residents who worked in refugee transit centres, detected in Austria in 2015, between calendar weeks 29 and 47. The species isolated from the cluster cases, including one mixed infection, were S. sonnei (n = 13), S. flexneri (n = 10) and S. boydii (n = 1). Eleven of 18 tested isolates were extended spectrum beta-lactamase (ESBL)-positive, including five of six ciprofloxacin-resistant and three azithromycin-resistant isolates.

A norovirus GII.P21 outbreak in a boarding school, Austria 2014.

OBJECTIVES: An Austrian boarding school reported a cluster of gastroenteritis on January 10, 2014. Environmental swabs from the school cafeteria and a nearby kebab restaurant tested positive for norovirus. The outbreak was investigated to identify its source(s). METHODS: An outbreak case was defined as a student or staff member with diarrhoea or vomiting that developed between January 7 and 13. Details on food exposure were collected via a self-administered questionnaire; risk ratios (RR) and 95% confidence intervals (CI) were calculated. Norovirus from the stool specimens of cases and asymptomatic kebab restaurant workers were genotyped. RESULTS: Twenty-eight cases were identified among 144 persons (attack rate 19%). The outbreak emerged and peaked on January 9, and ended on January 12. Compared to those who did not eat kebab, those who ate kebab on 7, 8, and 9 January were respectively 11 (95% CI 4.2-28), 6.7 (95% CI 3.4-13), and 9.3 (95% CI 4.0-22) times more likely to develop disease within the following 2 days. Stool specimens from three cases and three restaurant workers were positive for norovirus GII.P21. CONCLUSIONS: The kebab prepared by norovirus-positive restaurant workers was the most likely source of the outbreak. It is recommended that food handlers comply strictly with hand hygiene and avoid bare-handed contact with ready-to-eat food to minimize the risk of food-borne infection.

A Food Handler-Associated, Foodborne Norovirus GII.4 Sydney 2012-Outbreak Following a Wedding Dinner, Austria, October 2012.

On October 12, 2012, the provincial public health directorate of Salzburg reported a suspected norovirus (NV) outbreak among guests of a wedding-reception. The investigation aimed to confirm the causative agent, to identify the mode of transmission and to implement appropriate preventive measures. A probable outbreak case was defined as a wedding guest with diarrhoea or vomiting with disease onset from 7 to 10 October 2012 and who consumed food at the wedding dinner prepared by a hotel in the province Salzburg on 6 October 2012. A confirmed outbreak case fulfilled the criteria of a probable outbreak case and had a laboratory-confirmed NV infection. We conducted a cohort-investigation among the wedding guests. The case definitions were fulfilled in 26 wedding guests (25 %) including 2 confirmed cases. Females were 3.2 times more likely to develop disease (95 % CI 1.4-7.2) as compared to males. A mushroom dish was found to be associated with disease risk among females (risk ratio 2.3, 95 % CI 1.2-4.3). Two of 2 tested case-patients and 6 of 14 kitchen workers tested were positive for NV GII.4 Sydney. One kitchen staff-member worked during the wedding dinner despite diarrhoea. No food safety training was documented for the employees and the kitchen staff's restroom was lacking operational facilities for hand hygiene. We report the first investigated outbreak due to GII.4 Sydney, which was likely due to a symptomatic kitchen worker. Gender-specific eating behaviour may have posed female guests at higher risk of NV infection.

A food-borne outbreak of Shigella sonnei gastroenteritis, Austria, 2008.

We report on the first foodborne outbreak of Shigella sonnei described in Austria. On July 14 2008, AGES was informed of a cluster of 22 laboratory-confirmed cases of infection with S. sonnei restricted to public health district X in the province of Salzburg. All cases had attended a youth-group trip to a small village in the province of Tyrol from July 7 to July 9. An outbreak case among the trip participants was a person who (1) attended the trip and (2) fell ill with diarrhea in the period between July 8 and July 12. Among the 61 trip participants, 42 fitted the outbreak case definition, including 31 culture-confirmed cases. A household outbreak case was a person who (1) did not participate in the trip, (2) fell ill with diarrhea not before July 10 and (3) had household contact with an outbreak case between one and three days before onset of illness. Of the 11 household outbreak cases, 10 were tested by stool culture and four of these had a laboratory-confirmed S. sonnei infection. In addition, one of the two hostel staff tested positive for S. sonnei. All 36 isolates were biotype gl, lysotype 12, and were indistinguishable from each other by PFGE. A cohort study among the trip participants revealed a risk ratio of 2.71 for consumption of salad (on at least 1 of the 3 days of the trip) (95% CI: 1.38-5.32, P = 0.004). Among the 42cases, 34 could be explained by consumption of salad. The landlady of the hostel, who prepared the salad with bare hands, was a carrier and was assumed to be the source of contamination of the salad. Without proper epidemiological analytical investigation of this shigellosis outbreak, its association with food consumption would not have been identified.

Multiple-locus variable-number tandem repeat analysis for subtyping of Salmonella enterica subsp. enterica serovar Enteritidis.

Salmonella enterica subsp. enterica serovar Enteritidis is a major food-borne pathogen that caused most of Salmonella infections worldwide. S. Enteritidis phage type 4 (PT4) especially presents a real challenge for the classical typing methods. We developed a simple multiple-locus variable-number tandem repeat analysis (MLVA) assay based on three hypervariable variable-number tandem repeat (VNTR) loci for subtyping of Salmonella Enteritidis. Testing an arbitrary chosen strain collection of 110 S. Enteritidis isolates, comprising PTs 4, 8, and 21, the MLVA assay yielded a higher discriminatory power, corresponding to a Simpson's index of diversity (ID) of 0.91, when compared to pulsed-field gel electrophoresis (PFGE) which had a Simpson's ID of 0.41. To simplify interpretation of results, we developed a VNTR allele code based on the repeat unit number. This code can easily be exchanged. In conclusion, MLVA is a promising new tool to investigate outbreaks of S. Enteritidis and constitutes a useful addition to the current phage typing scheme.

Lessons learned from a Salmonella enteritidis phage type 4 outbreak in Austria, 2005.

An outbreak of gastroenteritis due to Salmonella Enteritidis phage type 4 occurred in people who attended a traditional hunting festivity in a small village in western Austria 6 through 11 November 2005. Of approximately 250 attendees, 227 had consumed dishes offered at the festival, and of these consumers 35 persons fulfilled the outbreak case definition (attack rate of 15.4%). Spätzle (traditional pastalike side dish) was most likely the contaminated part of the incriminated main course (relative risk of 18.9, 95% confidence interval of 4.6 to 76.7; P < 0.001). Thirteen eggs that remained from the preparation of the spätzle were negative for Salmonella when tested individually without shell disinfection, as were 1200 eggs collected at the egg production plant and examined with shell disinfection. The back-traced egg production farm had been initially certified as Salmonella free by a voluntary quality control program. However, an intensified environmental investigation of the incriminated egg production farm performed in the first quarter of 2006 and based on an appropriate method of sampling revealed Salmonella Enteritidis phage type 4 in 4 of 13 flocks. Although a combination of epidemiological and microbiological investigations allowed elucidation of the mode of spread, no restrictions were placed on the incriminated flocks of laying hens. These flocks were kept in production until they were stalled out due to age in August 2006. In June 2006, a cluster of 23 cases of Salmonella Enteritidis phage type 6 infection was again associated with this egg production farm. Evidence provided by epidemiological analyses is often disregarded by decision makers. However, negative results from microbiological testing of food involved in an outbreak are often weighted as strong evidence against a causal association between that food and the outbreak.

An outbreak of Norovirus infection affecting an Austrian nursing home and a hospital.

On November 9, 2004, a resident in a nursing home experienced a severe episode of vomiting in the dining room, in the presence of most of the other residents and members of staff. Following that episode, 17 of the 23 (73.9%) other residents and 7 of the 18 (38.9%) staff members fell ill with diarrhea and/or vomiting in the period up to November 17. A second cluster of gastroenteritis occurred between November 11 and 28, 2004, in a nearby hospital to which eight cases among the nursing home residents had been referred. Ten of 46 (21.7%) other hospital patients and 18 of 60 (30%) members of the hospital staff suffered from vomiting or diarrhea. Epidemiological and laboratory investigations proved a causal relationship between the two institutional clusters of short-lived gastroenteritis related by time and place, and identified Norovirus genotype GGII.4 (Jamboree-like) as the causative pathogen. Control measures for Norovirus, based on epidemiological and clinical features of the outbreak, were effectively implemented in the nursing home without waiting for virological confirmation. At the hospital, specific measures were not implemented until after virological confirmation of the causative agent, by which time 16 cases had already occurred. In a suspected Norovirus outbreak it is of great importance -- especially within closed and semiclosed settings -- to implement control measures as soon as possible, even before laboratory confirmation of the agent.