Source: UNIVERSITY OF MASSACHUSETTS submitted to NRP
EVALUATING THE RISK OF DEVELOPING NOROVIRUS VARIANTS WITH ENHANCED RECALCITRANCE TO DISINFECTANTS AS A CONSEQUENCE OF IMPROPER SANITATION PRACTICES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1030779
Grant No.
2023-67011-40346
Cumulative Award Amt.
$180,000.00
Proposal No.
2022-11414
Multistate No.
(N/A)
Project Start Date
Jun 1, 2023
Project End Date
May 31, 2026
Grant Year
2023
Program Code
[A7101]- AFRI Predoctoral Fellowships
Recipient Organization
UNIVERSITY OF MASSACHUSETTS
600 SUFFOLK ST FL 2 S
LOWELL,MA 01854-3983
Performing Department
Dept: Food Sciences
Non Technical Summary
Though food service establishments are typically required to follow minimum sanitation requirements by state, restaurants and food service applications are still responsible for 81% of food preparation reported outbreaks in the United States. Along with this, poor cleaning and sanitation are leading contributors to the incidence of foodborne outbreaks. More than half of norovirus outbreaks (60%) can be traced to infected people or carrier of norovirus that did not use proper sanitation practices during the preparation and serving of food.Implementation of proper sanitation practices is time-consuming and costly for many processors to implement, thus, we have seen an uprise in norovirus concernsNumerous properties of noroviruses make them difficult to control, including their low infectious dose and ability to persist on surfaces for weeks to months.Additionally,the active ingredients of many commonly used disinfectants have displayed some limitations in inactivating human noroviruses. Further, these disinfectants are often not properly applied--for example, improper use of chemicals and disinfectants,or application of the agent for a shorter contact time than recommended.Hypochlorite, quaternary ammonium and peroxyacetic acid compound solutions are among the most commonly used active ingredients in food service applications. Numerous studies suggest that quaternary ammonium (QUAT) compounds lack ideal efficacy against noroviruses and their surrogates, but they are one of the most commonly used disinfectants in food service settings. There is a lack of studies related to the development of enhanced norovirus resistancewhen repeated sub-fatal exposure to disinfectants, which we address in this work. Peroxyacetic acid (PAA), is another commonly utilized inactivation agent in food preparation/service. Numerous reports suggest that PAA is can be relatively effective against norovirus; however, contact times greater than one minute must be maintained.Though previous work has indicated a correlation between Good Manufacturing Practice (GMP) compliance and lower instances of norovirus contamination amongst food service applications, variability amongst state and federal requirements impacts the effectiveness of overall food service compliance in practice.Sanitation requirements for bacterial contaminants are often followed, even though it is often more difficult to inactivate nonenveloped viruses. It has been reported that as high as 30% of norovirus cases display no symptoms, which puts a higher emphasis on frequent and effectivecleaning and disinfection procedures to mitigate contamination from workers not displaying immediate symptoms.The research objective of this proposal is to evaluate and understand the potential of Tulane virus (a norovirus surrogate) to develop enhanced recalcitrance to serial sub-fatal treatments by different inactivation agents commonly used in food production and service settings.Further, the potential for the development of enhanced recalcitrance with norovirus surrogates against a number of different commonly used inactivation agents has not been investigated: peroxyacetic acid (food-grade inactivation agent, disinfectants).Our central hypothesis is that if serial sub-fatal treatments of these inactivation agents are applied, then virus variants with significantly enhancedresistanceto these inactivation agents (sanitizers)willdevelop.We believe this overall research design will provide a more global perspective of the potentialofsubfataltreatmentswiththeseinactivationagentstoresultinnorovirusvariantswith enhanced recalcitrance. These findings will be communicated through a risk model that shows routes of contamination of norovirus based on observed practices, and workshop training for food service workers to communicate the importance of norovirus awareness.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71250101101100%
Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
5010 - Food;

Field Of Science
1101 - Virology;
Goals / Objectives
Objective 1. Concealed observation of members within food service establishments to assess the current practices of cleaning and disinfectant techniques using approved sanitizers (research and extension).Objective 2. Investigate the potential for Tulane virus to develop enhanced recalcitrance to serial sub-fatal disinfectant treatments (quaternary ammonium and PAA), as well as the potential evolutionary mechanisms by which such enhanced recalcitrance occurs if observed (research)Objective 3. Develop a risk model that can be used as a communication method for understanding norovirus risk in a food retail environment.
Project Methods
1. GMP and sanitation observations will be made and discreetly recorded in a lab notebook regarding hand-washing occurrence, disinfectant used, aptitude about norovirus, disinfectant concentration, and disinfectant contact time. This data will serve the laboratory experimental parameters for viral resistance needed to create an accurate risk model. Human subjects will be studied in large foodservice environments in Massachusetts.2. Virus suspension assays will be performed based on ASTM E 1052-96, except disinfectant will be used against Tulane virus. Initially, inactivation agent will be diluted and different treatments times tested. A treatment time found to yield ~2-3 log reduction of virus (~1-2 log PFU remaining) will be selected for each virus and used for each cycle. Untreated, neutralization, and no virus controls will also be used with each treatment assay. General susceptibility of virus pools from cycles 1, 5, and 10 will be treated in suspension with disinfectant at a range of times and general virus susceptibility compared; with the evolution process being repeated at least thrice.From selected cycles of treated and untreated virus, virus isolates will be identified using plaque purification.3.Results of recalcitrance studies and observed practices will inform modeling of the risk of norovirus variants with enhanced recalcitrance introduction and contamination in food service settings. Risk models will cater to the stakeholders within the food service industry to ensure that the experiments are relevant and practical. Modeling tools provide a visual representation of norovirus introduction risks, and can be used to communicate active strategies for mitigating these hazards.

Progress 06/01/23 to 05/31/24

Outputs
Target Audience:The target audience for the work completed during this project period includes other researchers studying concealed observations of food safety behavior,viralrecalcitrance, and cell culture, particularly those who focus on sanitation outreach programming and risk assessment for retail food establishments. It should be particularly relevant to sanitation chemical companies, as such work will help to inform label accuracy for chemicals used to treat norovirus in food retail settings.It is also highly relevant to food producers and processors, as it could eventually allow for more conversations and awareness of norovirus resistance to chemicals. Changes/Problems:Our first major change is that our original plan was to conduct concealed observations under the guise of an "intern" position, without letting any members of the retail establishment know that I was conducting research. This has since been modified to include a broad announcement of the study to those who may come into contact with me in the retail setting. We have also developed a debriefing form, an updated agreement form to be signed by our stakeholder group, and Union agreement.Approval from the IRB has been prolonged due to these modifications in the protocol, thus delaying concealed observations. However, this has given me time to develop a robust protocol for recalcitrance detection, begin preliminary bench work, and derive a backlog of viral and cell stocks. What opportunities for training and professional development has the project provided?I havehad the opportunity to present myresearch at multiple local, national, and international meetings in the past year. Notably,Iwas able to present mywork at the 2023 International Association for Food Protection (IAFP) Annual Meeting in the form of a poster presentation, and am scheduled to give a technical talk on my research at IAFP 2024. I have also presented my work at the Northeast Center to Advance Food Safety (NECAFS) Annual Meeting, and at the Pioneer Valley Microbe Symposium.Additionally, I have had the opportunity to supervise twograduate researchers in the Kinchlalab who is working on a project related to that funded by the NIFA fellowship. I have also gained valuable professional experience while working with the UMass IRB, professionals in concealed observation work, and sanitation experts. How have the results been disseminated to communities of interest?Preliminary results for this project have been shared through poster presentations at scientific conferences from the regional to international level. Additionally, a research paper detailing the work completed toward objectives 1 and 2 of this project has been drafted and is being peer-reviewed by colleagues.Two extension outputs in the form of an online and in-person program have beencompleted toward objective 1, and are currently in peer-review. Additionally, discussions with IRB that will significantly impact future work in thefield of concealed observations have been shared with retail upper management at UMass (stakeholders for this study). What do you plan to do during the next reporting period to accomplish the goals?I have begun to develop a template for navigating the IRB for concealed observation work, which will significantly benefit future studies in the field. In the next year, I plan to complete IRB review and begin my concealed observations (objective 1), continue to collect data on Tulane recalcitrance (objective 2), interpret data found from recalcitrance studies (objective 2). This work will then inform the next stage of my study, where I will adjust bench conditions based on my observations, which will give me a better illustration of my risk model (objective 3).

Impacts
What was accomplished under these goals? My first objective was to conduct concealed observation of members within food service establishments to assess the current practices of cleaning and disinfectant techniques using approved sanitizers, which use research and extension techniques. To accomplish this goal, Ihave first initiateda thorough protocol reviewwith the UMass Internal Review Board (IRB). Since this work is novel to the field, I have navigated a review with the full review board to assess risks to stakeholders andworked with the food service union at our university to navigate resources for risks encountered through this work. I have also met with upper management of our retail dining sectors at UMass to discuss a plan for IRB compliance during the study, and professionals from past concealed observation studies around the country. Our IRB protocol is currently in review, as I have added new documents to the protocol, such as a debriefing form. I expect the protocol to be reviewed by early summer. My second objective is to investigate the potential for Tulane virus to develop enhanced recalcitrance to serial sub-fatal disinfectant treatments (quaternary ammonium and PAA), as well as the potential evolutionary mechanisms by which such enhanced recalcitrance occurs if observed. To accomplish this goal, recalcitrance protocol development and training have taken place in UMass laboratory facilities. Since Tulane virus needs a host cell line to grow, I have been working to grow and develop a library of cell and virus stocks to sustain our repeated sanitizer exposure over multiple trials. We have shifted the protocol to primary focus on PAA rather than both Quaternary Ammonium andPAA, due to PAA's popularityin retail dining sectors over Quats. Our preliminary studies on recalcitrance behavior are ongoing, and I expect to begin data analysis in the coming months. My third objective is to develop a risk model that can be used as a communication method for understanding norovirus risk in a food retail environment. To accomplish this goal, I have met with risk model experts in the food safety field to determine specific parameters and details that need to be accounted for in my concealed observations. I have worked with data science experts to determine risk modeling software for this work, and have developed a preliminary sketch of the model outputs.

Publications

  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Allingham, Christina, L.; Bolton, Jason; Machado, Robson; Johnston, Lynette; Cotter, Stephanie; Stevenson, Clinton; Yamamoto, Julie; Kinchla, Amanda J. (2024). Identification of Sanitation Job Tasks and Subsequent Training Requirements for Small Food Manufacturers Using a Modified Delphi Approach. Food Protection Trends
  • Type: Theses/Dissertations Status: Under Review Year Published: 2024 Citation: Christina Allingham, Amanda Kinchla, Matthew D. Moore,(2024). Developing Preventive Controls for Human Foods Prerequisite Education Trainings for Qualified Exempt Food Processors.
  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Allingham, Christina, L.; Bolton, Jason; Machado, Robson; Johnston, Lynette; Cotter, Stephanie; Stevenson, Clinton; Yamamoto, Julie; Kinchla, Amanda J. (2024). Lessons learned on developing an interdisciplinary sanitation program for small food producers focused on hybrid learning. Food Protection Trends
  • Type: Theses/Dissertations Status: Under Review Year Published: 2024 Citation: Allingham, Christina, L.; Bolton, Jason; Machado, Robson; Johnston, Lynette; Cotter, Stephanie; Stevenson, Clinton; Yamamoto, Julie; Kinchla, Amanda J. (2024). Developing a Skill-Building In-Person Sanitation Training Program for Food Entrepreneurs.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2024 Citation: Allingham, C.L., Kinchla A.J., Machado, R., Stevenson, C., Cotter, S., Yamamoto, J., Johnston, L., Nicholas, K., Hands-On Sanitation Programming Development for Small Food Processors, Technical Talk, IAFP Annual Meeting, Upcoming: July 2024
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Allingham, C.L., Kinchla A.J., Machado, R., Stevenson, C., Cotter, S., Yamamoto, J., Johnston, L., Nicholas, K., Hands-On Sanitation Programming Development for Small Food Processors, Poster, Pioneer Valley Microbe Symposium, Amherst, MA, March 2024
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Allingham, C.L., Kinchla A.J., Machado, R., Stevenson, C., Cotter, S., Yamamoto, J., Johnston, L., Nicholas, K., Hands-On Sanitation Programming Development for Small Food Processors, Poster, NECAFS Annual Meeting, January 2024
  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Fitzsimmons J., Allingham, C.L., Kinchla A.J., The Cost Barrier for Small and Medium Processor Compliance with Preventive Controls for Human Foods  Food Control (Submitted, In Review)
  • Type: Other Status: Under Review Year Published: 2024 Citation: Sanitation Controls Practitioners Program (SCPP) Part I: Online  Cotter, Stephanie; Yamamoto, Julie; Allingham, Christina; Kinchla, Amanda; Stevenson, Clint; Machado, Robson; Johnston, Lynette; Nicholas, Kate. (In peer review, NECAFS Clearinghouse).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Advances in the Detection and Control of Foodborne Viruses. Future of Food Symposium. McGill University and the Consortium de Recherche et innovations en Bioproc�d�s industriels au Qu�bec (CRiBiQ). Montreal, Quebec, Canada. 5/16/2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore Lab: Applied and Environmental Virology. UMass Food Science Advisory Board Meeting. Amherst, MA, USA. 4/19/2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Allingham, C.L., Kinchla A.J., Machado, R., Stevenson, C., Cotter, S., Bolton, J., Yamamoto, J., Johnston, L., Hybrid Sanitation Programming for Small Processors Guided By Industry Feedback, Poster, IAFP Annual Meeting, July 2023
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore Lab: Applied and Environmental Virology. Department of Defense Army Combat Feeding Division Soldier Center. Natick, MA, USA. 2/27/2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Developments in detection and control of highly transmissible viral pathogens. World Society for Virology 2023 Meeting. Riga, Latvia. 6/16/2023.
  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Christina Allingham, Miyu Taniguchi, Amanda Kinchla, Matthew D. Moore,(2024). The Influence of Simulated Organic Matter on the Inactivation of Viruses: A Review. Viruses