Fecal viral load and norovirus-associated gastroenteritis

We report the median cDNA viral load of norovirus genogroup II is >100-fold higher than that of genogroup I in the fecal specimens of patients with norovirus-associated gastroenteritis. We speculate that increased cDNA viral load accounts for the higher transmissibility of genogroup II strains through the fecal-oral route.     

Characterization of the norovirus 3C-like protease

The recombinant 3C-like protease of Chiba virus, a Norovirus, expressed in Escherichia coli cells was purified and characterized as to effects of pH, temperature, salt contents, and SH reagents on its proteolytic activity. The optimal pH and temperature of the 3C-like protease for the proteolytic activity were 8.6 and 37 °C, respectively. Increased concentration (100 mM) of monovalent cations such as Na⁺ and K⁺ was inhibitory to the activity. Hg²⁺ and Zn²⁺ remarkably inhibited the protease activity, while Mg²⁺ and Ca²⁺ had no virtual effect. Several sulfhydryl reagents such as p-chloromercuribenzoic acid, methyl methanethiosulfonate, N-ethylmaleimide and N-phenylmaleimide also blocked the activity, confirming the previous result that cysteine residue(s) were responsible for the proteolysis.     

Relationship between enteric pathogens and acute gastroenteritis disease severity: a prospective cohort study

Objectives

To evaluate the relationship between individual bacterial and viral pathogens and disease severity.

Vinyl-Stilbene Inhibits Human Norovirus RNA Replication by Activating Heat-Shock Factor-1

Norovirus (NV) is the most common cause of viral gastroenteritis, with the potential to develop into a fatal disease in those who are immuno-compromised, and effective vaccines and treatments are still non-existent. In this study, we aimed to elucidate the molecular mechanism of the previously identified NV replication inhibitor utilizing a vinyl-stilbene backbone, AC-1858. First, we confirmed the inhibition of the NV RNA replication by a structural analog of AC-1858, AC-2288 with its exclusive cytoplasmic subcellular localization. We further validated the induction of one specific host factor, the phosphorylated form of heat shock factor (HSF)-1, and its increased nuclear localization by AC-1858 treatment. Finally, we verified the positive and negative impact of the siRNA-mediated downregulation and lentivirus-mediated overexpression of HSF-1 on NV RNA replication. In conclusion, these data suggest the restrictive role of the host factor HSF-1 in overall viral RNA genome replication during the NV life cycle.     

Healthcare-associated foodborne outbreaks in high-income countries: a literature review and surveillance study, 16 OEDC countries, 2001 to 2019

BackgroundHealthcare-associated foodborne outbreaks (HA-FBO) may have severe consequences, especially in vulnerable groups.AimThe aim was to describe the current state of HA-FBO and propose public health recommendations for prevention.MethodsWe searched PubMed, the Outbreak Database (Charité, University Medicine Berlin), and hand-searched reference lists for HA-FBO with outbreak onset between 2001 and 2018 from Organisation for Economic Co-operation and Development (OECD) countries and HA-FBO (2012–2018) from the German surveillance system. Additionally, data from the European Food Safety Authority were analysed.ResultsThe literature search retrieved 57 HA-FBO from 16 OECD countries, primarily in the US (n = 11), Germany (n = 11) and the United Kingdom (n = 9). In addition, 28 HA-FBO were retrieved from the German surveillance system. Based on the number of outbreaks, the top three pathogens associated with the overall 85 HA-FBO were Salmonella (n = 24), norovirus (n = 22) and Listeria monocytogenes (n = 19). Based on the number of deaths, L. monocytogenes was the main pathogen causing HA-FBO. Frequently reported implicated foods were ‘mixed foods’ (n = 16), ‘vegetables and fruits’ (n = 15) and ‘meat and meat products’ (n = 10). Consumption of high-risk food by vulnerable patients, inadequate time-temperature control, insufficient kitchen hygiene and food hygiene and carriers of pathogens among food handlers were reported as reasons for HA-FBO.ConclusionTo prevent HA-FBO, the supply of high-risk food to vulnerable people should be avoided. Well working outbreak surveillance facilitates early detection and requires close interdisciplinary collaboration and exchange of information between hospitals, food safety and public health authorities.     

Viral gastrointestinal infections and norovirus genotypes in a paediatric UK hospital, 2014–2015

BackgroundDiarrhoea in children is a common disease; understanding the incidence of causative viruses can aid infection control and vaccine development.ObjectivesDescribe the incidence and characteristics of gastroenteric viruses including norovirus genotypes in a paediatric hospital cohort.Study designNorovirus, adenovirus, sapovirus, astrovirus, rotavirus qPCR and norovirus genotyping results for all stool specimens (n = 4786; 1393 patients) at a UK paediatric tertiary referral hospital June 2014–July 2015.Results and discussion24% (329/1393) of patients were positive for a GI virus; the majority were positive for norovirus (44%, 144/329) or adenovirus (44%, 146/329). The overall incidence of rotavirus (2%) is reduced compared to pre-vaccination studies; however the incidence of other GI viruses has not increased. Norovirus infections had a significantly higher virus burden compared to other GI viruses (P ≤0.03); sapovirus infections had the lowest viral burden.The number of norovirus cases per month did not follow the typical winter seasonal trend of nationally reported outbreaks. The number of cases per month correlates with the number of hospital admissions (R = 0.703, P = 0.011); the number of admissions accounts for 50% of the variability in number of cases per month. The breadth of genotypes seen (48% non-GII.4), suggests a community source for many norovirus infections and has implications for vaccine development.All GI viruses caused chronic infections, with the majority (50–100%) in immunocompromised patients. Incidence or duration of infection in chronic norovirus infections did not differ between genotypes, suggesting host-mediated susceptibility.     

A comparative evaluation of the sensitivity of two automated and two manual nucleic acid extraction methods for the detection of norovirus by RT-PCR

The aim of the study was to compare the sensitivity of a norovirus RT-PCR method using two manual RNA extraction methods (Qiagen and Roche) and two automated RNA extraction methods (Qiagen and Corbett). All four RNA extraction methods gave similar sensitivities although the automated methods, especially the Corbett, required significantly less labour than the manual methods. The automated methods also enabled RNA extraction of approximately two to three times the number of specimens in a given time period compared to manual methods.