Australian National Enterovirus Reference Laboratory annual report, 2022.

Abstract: Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System, and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2022, no cases of poliomyelitis were reported from clinical surveillance and Australia reported 1.69 non-polio AFP cases per 100,000 children, thereby meeting the WHO's performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A2, coxsackievirus A6, coxsackievirus A10, echovirus 18, enterovirus A71 and enterovirus C96 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus and environmental surveillance to complement the clinical system focussed on children. In 2022, thirty cases of wild poliovirus were reported from three countries (Afghanistan, Mozambique and Pakistan); 24 countries also reported cases of poliomyelitis due to circulating vaccine-derived poliovirus.

Changes in norovirus incidence in Victoria, Australia, during the COVID-19 pandemic, 2020-2021.

There were 142 norovirus positive outbreaks in Victoria for the 2020-2021 calendar years; however, almost half of these (48.6%) occurred in Q1 (January-March) of 2021. For the two-year period, 69.0% of all norovirus positive outbreaks were in childcare settings, and the predominant genotype was GII.P16/GII.2 (64.9%) followed by GII.P31/GII.4_2012 (20.9%). Norovirus incidence was particularly low in 2020 (n = 26) and close to average in 2021 (n = 116), but genotype diversity was low in both years. With the thought that 2022 will approach a more normal aspect to socialising and travel, norovirus incidence in 2022 may be predicted to increase above typical levels.

Australian National Enterovirus Reference Laboratory annual report, 2021.

Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System, and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2021, no cases of poliomyelitis were reported from clinical surveillance and Australia reported 1.31 non-polio AFP cases per 100,000 children, thereby meeting the WHO's performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A4, coxsackievirus A10, coxsackievirus A13 and enterovirus A71 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus and environmental surveillance to complement the clinical system focussed on children. In 2021, there were five cases of wild poliovirus reported from the two remaining endemic countries: Afghanistan and Pakistan. Including Afghanistan and Pakistan, 22 countries also reported cases of AFP due to circulating vaccine-derived poliovirus.

Australian National Enterovirus Reference Laboratory annual report, 2020.

ABSTRACT: Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2020, no cases of poliomyelitis were reported from clinical surveillance; Australia reported 1.09 non-polio AFP cases per 100,000 children, thereby meeting the WHO's performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A10 and coxsackievirus A16 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus surveillance and environmental surveillance to complement the clinical system focussed on children. In 2020, there were 140 cases of wild poliovirus reported from the two remaining endemic countries: Afghanistan and Pakistan. Another 28 countries reported cases of circulating vaccine-derived poliovirus.

Global Trends in Norovirus Genotype Distribution among Children with Acute Gastroenteritis.

Noroviruses are a leading cause of acute gastroenteritis (AGE) among adults and children worldwide. NoroSurv is a global network for norovirus strain surveillance among children <5 years of age with AGE. Participants in 16 countries across 6 continents used standardized protocols for dual typing (genotype and polymerase type) and uploaded 1,325 dual-typed sequences to the NoroSurv web portal during 2016-2020. More than 50% of submitted sequences were GII.4 Sydney[P16] or GII.4 Sydney[P31] strains. Other common strains included GII.2[P16], GII.3[P12], GII.6[P7], and GI.3[P3] viruses. In total, 22 genotypes and 36 dual types, including GII.3 and GII.20 viruses with rarely reported polymerase types, were detected, reflecting high strain diversity. Surveillance data captured in NoroSurv enables the monitoring of trends in norovirus strains associated childhood AGE throughout the world on a near real-time basis.

Decreased incidence of enterovirus and norovirus infections during the COVID-19 pandemic, Victoria, Australia, 2020.

ABSTRACT: Significant reductions in the incidence of enteroviruses and noroviruses, both transmitted primarily by the faecal-oral route, were noted in 2020 compared to the previous decade, in Victoria, Australia. The enterovirus specimen positivity rate was reduced by 84.2% in 2020, while the norovirus outbreak positivity rate declined by 49.0%. The most likely explanation for these reductions is the concurrence of social restrictions, physical distancing, personal hygiene awareness and international and domestic border closures in response to the COVID-19 pandemic.

GII.4 norovirus recombinant causes gastroenteritis epidemic in Eastern Australia, 2017.

In Victoria, Australia, 160 gastroenteritis outbreaks were norovirus positive for the period January-September 2017. A distinctive peak in norovirus outbreaks was seen May-August, with 118 positive outbreaks occurring in the peak period. The peak was primarily due to the emergence of a GII.P4_NewOrleans_2009/GII.4_Sydney_2012 recombinant that had genetically changed sufficiently to escape herd immunity. This recombinant was also identified elsewhere in Australia, with highly similar sequences identified in Queensland during the same time period. The recombinant GII.P4_NewOrleans_2009/GII.4_Sydney_2012 has not been reported to cause norovirus epidemics outside Australia, suggesting regional factors play a role in determining norovirus genotype incidence.

The molecular epidemiology of norovirus outbreaks in Victoria, 2016.

Noroviruses are a leading cause of outbreaks of gastroenteritis. This study examined the incidence and molecular characteristics of norovirus outbreaks in Victoria, Australia in 2016. Norovirus was detected in 52.4% of gastroenteritis outbreaks surveyed and the year was notable in that there was no significant temporal peak in norovirus outbreaks. Norovirus Open Reading Frame (ORF)2 (capsid) genotypes were successfully identified in 84 of 110 norovirus outbreaks and included GI.3, GI.6, GI.9, GII.2, GII.3, GII.4, GII.6, and GII.17. Norovirus GII.4 was the most common ORF2 genotype detected (55.9%). Other relatively common ORF2 genotypes included GII.2 (19.0%), GII.17 (9.5%), GI.3 (7.1%) and GII.3 (4.8%). The GII.P4_NewOrleans_2009/GII.4_Sydney_2012 recombinant emerged as an important new GII.4 form. The study also confirmed the identity of three ORF1/ORF2 recombinant forms as follows: GII.P16/GII.2, GII.P16/GII.3 and GII.P16/GII.4. Statistical analysis indicated GII.4 (ORF2) was much more common in healthcare settings than in non-healthcare settings. The study indicates 2016 was a transition year in Victoria, Australia, in that the previous norovirus epidemic strain had diminished to the point where it was no longer dominant but as yet no replacement epidemic strain had become obvious.

Evolutionary changes in the capsid P2 region of Australian strains of the norovirus GII.Pe_GII.4.

PURPOSE: The protruding (P) 2 region of the norovirus capsid is thought to include hypervariable sites involved in receptor binding. This study examines the changes that occurred in the P2 region of GII.Pe_GII.4 norovirus in the course of its evolution from a precursor phase (2008-2009), to an intermediate phase (2010) and finally to an epidemic phase (2012-2015). METHODOLOGY: Twenty-two P2 region amino acid (aa) sequences (166 aa long) from all phases of the evolution of the virus were compared and the changes analysed.Results/key findings. Twenty sites in the P2 region underwent aa change and of these, 10 corresponded to previously proposed hypervariable sites and 10 to novel hypervariable sites. It was notable that aa changes at two sites, X and Y, only emerged as the epidemic phase progressed. 3D computer modelling of the P2 region indicated that neither X nor Y were in the uppermost 'crown', but further down in the 'neck' portion. The location of X and Y and the nature of aa change at Y suggest these sites were important in enhancing the structural integrity of the capsid, which in turn may have facilitated the longer term viability of the virus. CONCLUSION: The current study helps establish the validity of previously proposed hypervariable sites in the P2 region as well as indicating new ones. It also provides quantitative and qualitative data on how these sites changed over the evolutionary history of a particular norovirus strain.

An outbreak of foodborne norovirus gastroenteritis linked to a restaurant in Melbourne, Australia, 2014.

INTRODUCTION: In May 2014 an outbreak of norovirus occurred among patrons of a restaurant in Melbourne, Australia. Investigations were conducted to identify the infectious agent, mode of transmission and source of illness, and to implement controls to prevent further transmission. METHODS: A retrospective case-control study was conducted to test the hypothesis that food served at the restaurant between 9 and 15 May 2014 was the vehicle for infection. A structured questionnaire was used to collect demographic, illness and food exposure data from study participants. To ascertain whether any food handlers had experienced gastroenteritis symptoms and were a possible source of infection, investigators contacted and interviewed staff who had worked at the restaurant between 9 and 16 May 2014. RESULTS: Forty-six cases (including 16 laboratory-confirmed cases of norovirus) and 49 controls were interviewed and enrolled in the study. Results of the analysis revealed a statistically significant association with illness and consumption of grain salad (OR: 21.6, 95% CI: 1.8-252.7, P = 0.015) and beetroot dip (OR: 22.4, 95% CI: 1.9-267.0, P = 0.014). An interviewed staff member who reported an onset of acute gastrointestinal illness on 12 May 2014 had prepared salads on the day of onset and the previous two days. DISCUSSION: The outbreak was likely caused by person-to-food-to-person transmission. The outbreak emphasizes the importance of the exclusion of symptomatic food handlers and strict hand hygiene practices in the food service industry to prevent contamination of ready-to-eat foods and the kitchen environment.

The molecular epidemiology of norovirus outbreaks in Victoria, 2014 to 2015
.

Noroviruses are a leading cause of outbreaks of gastroenteritis. This study examined the incidence and molecular characteristics of norovirus outbreaks in healthcare and non-healthcare settings in Victoria, Australia, over 2 years (2014-2015). Norovirus was detected in 65.7% and 60.4% of gastroenteritis outbreaks investigated for the years 2014 and 2015 respectively. There was a significant decline in the number of norovirus outbreaks in the period 2014 to 2015 although in both years norovirus outbreaks peaked in the latter part of the year. Norovirus Open Reading Frame (ORF) 2 (capsid) genotypes identified included GI.2, GI.3, GI.4, GI.5, GI.6, GI.9, GII.2, GII.3, GII.4, GII.6, GII.7, GII.8, GII.13 and GII.17. GII.4 was the most common genotype detected. In addition, the following ORF 1/ORF 2 recombinant forms were confirmed: GII.P4_NewOrleans_2009/GII.4_Sydney_2012, GII.P12/GII.3, GII.Pb (GII.21)/GII.3, GII.Pe/GII.2 and GII.Pe/GII.4_Sydney_2012. A significant decline was noted in the chief norovirus strain GII.Pe/GII.4_Sydney_2012 between 2014 and 2015 but there was a re-emergence of a GII.P4_ NewOrleans _2009 norovirus strain. Outbreaks involving the GII.P17/GII.17 genotype were also detected for the first time in Victoria. GI genotypes circulating in Victoria for the 2 years 2014 and 2015 underwent a dramatic change between the 2 years of the survey. Many genotypes could occur in both healthcare and non-healthcare settings although GI.3, GII.6, and GII.4 were significantly more common in healthcare settings. The study emphasises the complex way in which norovirus circulates throughout the community.

A norovirus intervariant GII.4 recombinant in Victoria, Australia, June 2016: the next epidemic variant?

A norovirus recombinant GII.P4_NewOrleans_2009/GII.4_Sydney_2012 was first detected in Victoria, Australia, in August 2015 at low frequency, and then re-emerged in June 2016, having undergone genetic changes. Analysis of 14 years' surveillance data from Victoria suggests a typical delay of two to seven months between first detection of a new variant and occurrence of a subsequent epidemic linked to that variant. We consider that the current recombinant strain has the potential to become a pandemic variant.

The Comparative Molecular Epidemiology of GII.P7_GII.6 and GII.P7_GII.7 Norovirus Outbreaks in Victoria, Australia, 2012-2014.

The comparative molecular epidemiology of the related GII.P7_GII.6 and GII.P7_GII.7 noroviruses has not been examined in detail. ORF 1, ORF 2 and ORF 1/ORF 2 RT-PCR as well as sequencing and phylogeny analysis were carried out on faecal specimens from 873 gastroenteritis outbreaks in Victoria, Australia (2012-2014). There were 575 (66%) detected as positive for norovirus by means of ORF 1 RT-PCR and/or ORF 2 RT-PCR. Of these, 24 (4.2%) were GII.6 (ORF 2) outbreaks, 7 (1.2%) were GII.7 (ORF 2) outbreaks, and 1 outbreak (0.2%) involved both GII.6 (ORF 2) and GII.7 (ORF 2) noroviruses. The median age of patients identified with GII.6 (ORF 2) (84 years) was significantly different from that of patients identified with GII.7 (ORF 2) (39 years). ORF 2 GII.6 and ORF 2 GII.7 sequences were always associated with a GII.P7 ORF 1 sequence, and GII.P7 sequences fell into two clusters, with one corresponding to the GII.6 ORF 2 genotype and the other to the GII.7 ORF 2 genotype, thereby indicating that the ORF 1 has been evolving separately for the two viruses. Thus, two closely related noroviruses can have a markedly different incidence and epidemiology.

Emergence of GII.Pg norovirus in gastroenteritis outbreaks in Victoria, Australia.

The ORF 1 GII.Pg genotype represents an obligatory recombinant comprising the ORF 1 GII.Pg genotype and a number of ORF 2 genotypes. The emergence, incidence, and molecular features of GII.Pg norovirus have never been considered in detail and are the subject of the current study. Over the period 2002-2013, GII.Pg norovirus was detected in 16 outbreaks in Victoria, Australia. It was first identified in 2009 and thereafter was detected at low level in each year of the study. GII.Pg norovirus outbreaks occurred in both healthcare and non-healthcare settings and could be found in individuals with a broad range of ages. The seasonality of GII.Pg norovirus outbreaks was significantly different from that of all other (non-GII.Pg) norovirus outbreaks. For the 15 GII.Pg norovirus outbreaks where ORF 2 sequencing data were available, two ORF 2 recombinant genotypes were found: GII.1 in 5 (33%) outbreaks and GII.12 in 10 (67%) outbreaks. The ORF 1 phylogenetic tree shows that the GII.Pg ORF 1 genotype fell into two distinct groups. The ORF 2 phylogenetic tree indicates that the GII.1 and GII.12 clusters each corresponded to one of the groups in the ORF 1 tree. This indicates the two recombinant forms were evolving in parallel and not one from the other. Analysis of age data indicates the GII.1 and GII.12 recombinant forms circulated in different ways in the community. J. Med. Virol. 88:1521-1528, 2016. © 2016 Wiley Periodicals, Inc.

Electron microscope detection of tubular aggregates in the mosquito cell line C6/36 treated with Culex australicus (mosquito) homogenate.

'Tubular aggregates' are morphologically distinct cytoplasmic structures that have been linked to a variety of pathological conditions. This report documents the presence of tubular aggregates in an insect cell line (C6/36 cells derived from Aedes albopictus) following inoculation of the cells with material derived from cell culture passaged homogenized Culex australicus mosquitoes. The tubular aggregates were detected in ∼2% of treated cells and had three morphological forms that were termed primary, secondary and tertiary, with progressively greater levels of structural complexity. The findings indicate that tubular aggregates can be induced in an insect cell culture system by an unidentified agent present in some mosquitoes.

Chronic norovirus infection in a patient with a past history of Burkitt lymphoma.

The noroviruses are a leading cause of gastroenteritis worldwide. Although the illness is normally mild and self-limiting, there is a growing literature documenting the chronic excretion of norovirus in the immunocompromised. The aim of the current study was to examine the molecular features of chronic norovirus excretion in an immunocompromised patient with a past history of Burkitt lymphoma. During the 241 day course of the study from December 2013 to August 2014, seven faecal specimens were collected from the patient, tested for norovirus by RT-PCR and further analysed in the open reading frame (ORF) 1 and ORF 2 regions. All seven specimens were positive for norovirus by RT-PCR. Molecular sequencing in the polymerase (ORF 1) and capsid (ORF 2) regions indicated that the norovirus could be classified as GII.4 (2006b)/GII.4 (unknown). No significant mutation was found in the ORF 1 or ORF 2 regions analysed over the period of the study. The current report appears to be the first to document chronic norovirus excretion in a patient with a past history of Burkitt lymphoma. It is also the first to indicate long term norovirus excretion in a given individual need not involve major genetic change in key regions of the genome.

Evaluation of the updated RIDA®QUICK (Version N1402) immunochromatographic assay for the detection of norovirus in clinical specimens.

The sensitivity and specificity of the R-Biopharm RIDA(®)QUICK (N1402) immunochromatography assay for norovirus detection was examined using fecal material from Australian gastroenteritis incidents. The study involved the analysis of 3 groups of specimens; group 1 comprised 100 norovirus open reading frame (ORF) 1 RT-PCR positive specimens; group 2 comprised 100 ORF 1 RT-PCR norovirus negative specimens and group 3 comprised 12 specimens containing common gastroenteritis viruses other than norovirus. The RIDA(®)QUICK (N1402) assay detected both GI and GII norovirus and had an overall sensitivity of 87%. Genotype analysis of the capsid region of the genome (ORF 2) indicated the RIDA(®)QUICK (N1402) assay could detect a range of genotypes including GI.1, GI.2, GI.3, GI.4, GI.5, GII.3, GII.4 (including variants GII.4 (2009-like), GII.4 (2012), GII.4 (2012-like) and GII.4 (unknown)), GII.6, GII.13 and GII.21. The assay had good sensitivity for both GI and GII norovirus. The assay had a specificity of 97% and did not cross react with a number of common fecal viruses. However, one of eight rotavirus positive, norovirus negative specimens gave a positive result; rotavirus cannot be taken as the cause of such a false positive but cannot be excluded either. The kit was quick and easy to use and would be valuable in point-of-care testing.