New approaches for the quantitative detection of human pathogenic viruses within the freshwater-marine continuum

Lead Research Organisation: Centre for Environment, Fisheries and Aquaculture Science
Department Name: CEFAS Weymouth Laboratory

Abstract

Viruses pose one of the biggest threats to human wellbeing being responsible for numerous infections and millions of deaths worldwide each year. Most of these viral diseases are passed via the faecal-oral route in which contaminated food and water are frequently implicated in the primary infectivity phase. Although many of these infections are self-limiting, the societal and economic burden should not be underestimated. For example, Norovirus (NoV) is estimated to cause over 2 million cases of illness in the UK each year resulting in millions of days of lost productivity and an economic burden estimated to exceed £100 million to the NHS directly and over £2 billion annually to the wider economy. Worryingly, it is clear from a range of critical reviews that the burden of waterborne disease is likely to increase in Europe in response to climate change. This increasing problem is being exacerbated by increased pressure on wastewater infrastructure (due to population rise), sewer misconnections and a greater incidence of storms and flood events causing the release of untreated sewage (stormwater discharge) into river networks and the coastal zone. Considering the magnitude of the problem and the disease burden forecast for the near future, it is timely to develop new strategic approaches for mitigating against viral contamination and to develop new and improved risk assessment tools for protecting human health.

In view of this, our proposal aims to address the critical need to develop and validate new tools for the detection and surveillance of human pathogenic viruses in freshwater, estuarine and coastal environments. Specifically, we will design and test experimental and modelling tools to permit the robust recovery and quantification of viral populations from contrasting matrices (e.g. seawater, freshwater, sediments, effluent, shellfish). These tools will be designed to capture the viral populations in both space and time. We will focus on viruses of strategic importance from a human health perspective (e.g. Norovirus, Sapovirus, Hepatitis A/E), however, these will be placed in a wider context via metavirome analysis of RNA and DNA viral communities. These techniques will be deployed and demonstrated at the catchment-to-coast scale whilst simultaneously answering fundamental questions about the temporal and spatial dynamics of viral flow. This knowledge will be used to validate next generation mathematical models capable of predicting viral flow through the river network and coastal zone. Combined, this information will be used with key stakeholders (e.g. Cefas) in the implementation of new methods and guidelines for assessing infection risk (e.g. in recreational waters, beaches & shellfisheries) and for protecting human health.

Our proposal directly addresses the strategic aims of the NERC Environmental Microbiology and Human Health (EMHH) Programme. As requested by the call, we will provide "scientific evidence to support fast and efficient identification of pathogenic microorganisms in environmental media which can be used in appropriate tools and models for the protection of public health targeting the freshwater and coastal zone". The work is also directly relevant to the policy objectives and strategic aims of the Food Standards Agency, Defra and European Union (DG Sanco, and DG Mare).

Planned Impact

This proposal is designed to protect human health from the ever presence threat of viral infection that exists in the environment. For example, Norovirus is estimated to cause over 2 million cases of illness in the UK each year, resulting in millions of days of lost productivity and an economic burden estimated to exceed £100 million to the NHS directly and over £2 billion annually to the wider economy. A reduction in the disease incidence would therefore be of great benefit to the UK economy. To ensure our project has lasting impact, a range of key stakeholders have been actively engaged in the formulation and writing of this proposal to ensure that it directly meets their needs, complements existing research and is fit for purpose. Specifically, this proposal will be of benefit to water quality managers and regulators, both national and international. The main UK beneficiaries will be the Food Standards Agency, Cefas (Defra), Environment Agency, water companies, local authorities and the shellfish industry as well as UK academia. Internationally, the work will directly inform the European Environment Agency, EU-DG MARE and EU-DG SANCO, however, our project addresses an issue of global relevance, and therefore the impact will be far-reaching. Stakeholders will benefit as follows:

1. Human pathogenic viruses are rarely measured in river, estuarine and coastal waters and therefore are not used by the Environment Agency, Food Standards Agency or EU for regulatory purposes. Although ISO accredited methods of detection have been established by the team (e.g. NoV in shellfish by Cefas), uncertainty still remains on their detection in some matrices and additionally as to whether these PCR-detected viruses are still infective to humans. Therefore the formulation of regulatory standards for shellfish, bathing waters etc is currently a subject of considerable debate. As risk assessment must be based on robust scientific research, we will provide key underpinning data for policymakers and regulators.

2. Cefas work directly with the UK government and EU to monitor the marine environment and to demonstrate UK compliance with national and international marine legislation. A key part of Cefas' work is to detect and advise on the effects of anthropogenic drivers on the marine environment. Having Cefas embedded in the project will bring lasting benefits in ensuring that the information generated will have real impact. As an example, David Lees (Cefas) is Director of the European Union Reference Laboratory (EURL) monitoring bacteriological and viral contamination of bivalve molluscs which represents a primary forum for our findings. Our existing strong links with the UK Food Standards Agency and the European Union (Directorate-General for Maritime Affairs and Fisheries, DG-MARE; Directorate-General for Health and Consumer, DG-SANCO), the Consumers, Health and Food Executive Agency (CHAFEA), European Food Standards Agency (EFSA), European Centre for Environment and Human Health (ECEHH) means that we can disseminate and discuss the project findings with the regulatory and medical community through well-established channels.

3. Our project is highly relevant to water companies. Welsh Water are key stakeholders in our project while delivery to the wider water industry will be made through the UK Water Industries Research (UKWIR).

4. Key stakeholders are Public Health Wales, Public Health England and Scotland Public Health Network.

5. The Shellfish Association of Great Britain (SAGB) is an ideal vehicle to disseminate our findings to the shellfish industry and the aquaculture industry in general. Shelagh Malham attends regular SAGB meetings which bring together the shellfish industry including producers and processers, several government and non-governmental organisations and has lobby activity within Westminster for issues affecting the industry. This proposal addresses some of the top priority issues facing the industry.

Publications

10 25 50
 
Description The use of tools for quantifying the integrity of virus capsids are not a viable tool for infering virus infectivity.
Exploitation Route Several researchers are using inappropriate tools for infering viability of human norovirus in environmental samples. Future insights into viral infectivity in these samples would be better sought using surrogates for un-culturable viruses until a suitable direct culture method is developed.
Sectors Agriculture, Food and Drink,Environment

 
Title Most probable number (MPN) assay for detection of infectious genogroup II F specific RNA coliphage in bivalve shellfish molluscs 
Description This method provides a way to accurately quantify infectious genogroup II F specific RNA coliphage (FRNAP-II) in bivalve shellfish molluscs. This was developed as a replacement for the membrane hybridisation method previously used in our laboratory which was time consuming and labour intensive. The new method is based on growing FRNAP-II in broth culture in an MPN format and then detecting the presence of FRNAP-II by RT-PCR. Growth conditions, RNA extraction and RT-PCR have all been optimised to improve efficiency of the method. A protocol has now been developed in our laboratory for use by trained technicians. 
Type Of Material Technology assay or reagent 
Year Produced 2019 
Provided To Others? No  
Impact This method will be made available to others by a peer-reviewed publication which is currently being written. The detailed protocol will also be made available once the validation data have been fully analysed. 
 
Description Lisa Cross is an invited speaker at 'Soapbox Science' - a novel public outreach platform for promoting women scientists and the science they do. The event will take place in a public area in Exeter and transform it into an arena for public learning and scientific debate. 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact The event will take place on Saturday 24th of June 2017, the aim is to transform Exeter city centre into a hub of scientific learning and discussion, as female scientists take to their soapboxes to showcase science to the general public. The event's mission is to help eliminate gender inequality in science by raising the profile, and challenging the public's view, of women and science. I will be presenting on the topic of Infectious diseases - notably Norovirus.
Year(s) Of Engagement Activity 2017
URL http://soapboxscience.org/soapbox-science-2017-exeter
 
Description NERC Environmental Microbiology and Human Health Programme: Programme Integration Group (PIG), First Annual Programme Conference 16th Dec 2016 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Dr David Lees presented current research finding pertaining to work package 5: 'Establishing Viral Infectivity from Environmental Samples' at the inaugural meeting of the Environmental Microbiology and Human Health (EMHH) Programme Integration Group (PIG).
The EMHH Programme incorporates; the VIRAQUA project (tracing enteric viruses through the aquatic environment), the RESERVOIR project, identifying antimicrobial resistance in rivers and assessing the risks associated with specific pollutants and land management practices such as sewage sludge disposal. The RAMBIE and the ENDOTOX projects both focus on bioaerosols. The RAMBIE project is developing advanced tools for the rapid monitoring of Bioaerosols in urban, agricultural and industrial environments whilst ENDOTOX are developing new ways of characterising and quantifying emissions of endotoxin in the air at composting and farm sites modelling emissions and dispersion patterns.
NERC established the concept of the programme integration group in the hope that linking up certain projects would lead to a sharing of ideas and hopefully research innovation in those areas. The conference was well attended by stakeholders, project members and NERC staff and was a good opportunity for networking.
Year(s) Of Engagement Activity 2016
URL http://www.nerc.ac.uk/research/funded/programmes/emhh
 
Description Presentation at ISFEV conference in Arizona, 2018 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Presentation entitled: Towards routine analysis of infectious norovirus in bivalve shellfish
Given at the International Society for Food and Environmental Virology (ISFEV) conference in October 2018.
Year(s) Of Engagement Activity 2018
 
Description Water Microbiology Conference, Chapel Hill, North Carolina. KEYNOTE SPEAKER 'Improving health controls for viruses in Bivalve Molluscs' 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact KEYNOTE SPEAKER at Water Microbiology Conference, Chapel Hill, North Carolina. Title: 'Improving health controls for viruses in Bivalve Molluscs'
May 2016
Year(s) Of Engagement Activity 2016
URL http://waterinstitute.unc.edu/conferences/watermicro2016/speakers