Source: UNIV OF MASSACHUSETTS submitted to NRP
DEVELOPMENT AND EVALUATION OF MAGNETIC LIQUIDS AS AN EMERGING FOODBORNE PATHOGEN CONCENTRATION TOOL
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1032405
Grant No.
2024-67017-42443
Cumulative Award Amt.
$649,950.00
Proposal No.
2023-08788
Multistate No.
(N/A)
Project Start Date
Jun 1, 2024
Project End Date
May 31, 2027
Grant Year
2024
Program Code
[A1332]- Food Safety and Defense
Recipient Organization
UNIV OF MASSACHUSETTS
(N/A)
AMHERST,MA 01003
Performing Department
(N/A)
Non Technical Summary
Foodborne viruses are the leading cause of foodborne illness and fourth leading cause of foodborne death in the US.Foodborne viruses are also estimated to cause billions of dollars of losses annually. There are a number of challenges to controlling these viruses, and one of them is the ability to rapidly detect them in food and environmental samples in order to take measures to prevent their spread. Often, viral contamination of foods and the environment occurs at low levels that can still get people sick, but make the viruses difficult to detect. A problem with all food and environmental detection of viruses that infect animals is the fact that a cultural enrichment stepto grow the number of viruses in the sample and increase the ease of detecting them is not as feasible as for bacterial pathogens. Therefore, the viruses need to be picked out of large, complex food and environmental samples in order to be detected--a lot like finding and capturing needles in a haystack. However, traditional methods that capture virusesare cost-prohibitive and have other limitations the reduce the ease with which they can be utilized in-field or at point-of-preparation. Magnetic ionic liquids (MILs) and deep eutectic solvents (DESs) are a highly diverse group of hydrophobic liquids that have shown promise for concentration of bacteria and other biological targets in foods; however, their ability to concentrate viruses from foods and the environment has not been studied. MILs and DESs also have the advantage of being very stable, and only require a magnetic for capture, reducing the need for heavy or electrical equipment and making portable processing of foods feasible. The goal of the proposed project is to develop and evaluate these liquids as reagents for portable, rapid concentration of viruses from foods and the environment for downstream portable detection. Further, the proposed work will attempt to anchor specific molecules that specifically target viruses to MILs and determine if this can improve the concentration of viruses from foods. This would be a new paradigm in the use of MILs for capture of biological targets if successful.
Animal Health Component
45%
Research Effort Categories
Basic
10%
Applied
45%
Developmental
45%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71240301101100%
Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
4030 - Viruses;

Field Of Science
1101 - Virology;
Goals / Objectives
Rapid, portable detection of foodborne viruses is essential for their control. Although a number of promising portable technologies exist for downstream detection of foodborne viruses, their utilization in-field and at point of service is limited by the need for upstream concentration and purification techniques that are often time-consuming and not portable. Magnetic liquids are a diverse class of stable hydrophobic liquids with magnetic properties that have shown promise for concentration and detection of a number of biological targets, but have not been evaluated for foodborne viruses.The overall objective of the proposed work is to evaluate the ability of magnetic liquids to capture and concentrate viruses from foods and the environment, as well as develop and evaluate ligand-modified MILs for the same purpose. The ultimate goal of the work is to develop magnetic liquid-based reagents to serve for realistic, portable concentration of foodborne viruses (and other foodborne pathogens) that enables rapid detection of viruses in foods and the environment in food production and service settings.The specific objectives of the proposed work are:Objective 1: Develop and evaluate the ability of different magnetic ionic liquids (MILs) to capture and concentrate foodborne viruses (norovirus, hepatitis A virus) from food and environmental samples prior to detection. In this objective, we will evaluate the ability of different formulations of different types of magnetic ionic liquids (MILs; Co, Ni, Dy) to capture foodborne viruses (norovirus, hepatitis A virus) from buffer and relevant food matrices (leafy greens and berries).Objective 2: Develop and evaluate the ability of different deep eutectic solvents (DESs) to capture and concentrate foodborne viruses (norovirus, hepatitis A virus) from food and environmental samples prior to detection. In this objective, we will evaluate the ability of different formulations of different types of deep eutectic solvents to capture foodborne viruses (norovirus, hepatitis A virus) from buffer and relevant food matrices (leafy greens and berries).Objective 3: Develop and evaluate the ability of ligand-modified magnetic ionic liquids (nucleic acid aptamer) to capture and concentrate noroviruses. In this objective, we will formulate the best performing MILs to be modified with norovirus-specific nucleic acid aptamers and peptides for specific capture and concentration of norovirus, as well as evaluate their potential as concentration reagents for these viruses in foods commonly implicated with foodborne virus transmission (leafy greens and berries).
Project Methods
The project will be conducted largely as proposed in the submitted narrative. On a global level, we will evaluate the performance of a different group of magnetic ionic liquids (MILs) and deep eutectic solvents (DESs) to concentrate noroviruses and hepatitis A virus from buffer, then testing the ability to concentrate viruses from more complex food matrices (lettuce and berries). Similarly, we will conduct work to create MILs with a set of specific anti-virus ligands (aptamers) for specific capture of virus from buffer and then foods.Efforts:The performance and strengths and weaknesses of the magnetic liquids for concentration of the viruses from buffers and foods will be evaluated by graduate students and undergraduate research volunteers, who will be trained on how to synthesize magnetic liquids, concentrate, and detect viruses from buffers and food samples. The results of the work will be disseminated in multiple forums both through publications, poster presentations, technical talks, as well as guest lectures for different food science undergraduate/graduatecourses (like Food Microbiology Lecture).Evaluation:The success of the project will be evaluated by number of scientific products produced, students trained, student awards received, posters presented, technical talks given, and seminars/invited talks given. Specifically, the project will be considered successful if 3 articles/reviews are published in respected scientific journals, 5 poster/technical talk presentations are given, 4 invited talks/seminars are given,2 PhD students are graduated, and at least 2 undergraduate researchers are trained.Scientifically, the capture and recovery efficiency of thedifferent magnetic liquid formulations will be compared to other concentration methods previously reported, with consideration of the portability of the different methods and their cost. The project will be considered successful if the magnetic liquids display concentration efficiency comparable or greater than other concentration methods that have less portability.

Progress 06/01/24 to 05/31/25

Outputs
Target Audience:The target audience of this project is primarily other members of the food safety community. Specifically, other foodborne virologists and those working for safety testing at establishments of risk will be of interest. The work is also targeting other food safety researchers and environmental virologists and microbiologists. Because of a lack of efficient, portable virus concentration methods, testing for foodborne viruses in the food and service industry in environmental and food samples is quite low. The project is in part targeting itself to those in the industry with hopes foodborne virus testing will become more commonly conducted if a cheap, efficient concentration and detection assay that has been validated methodologically is provided. Changes/Problems:We have changed personnel so there is a delay in training for the new graduate student taking over at UMass. There also has been an issue getting the hepatitis A virus host cells to propagate, so we have ordered a new set of cells from ATCC. What opportunities for training and professional development has the project provided?The UMass student primarily involved with this project, Sloane Stoufer, has had the opportunity to present her research at multiple local, national, and international meetings. Additionally, Sloane has had the opportunity to supervise a former undergraduate researcher who went on the complete an accelerated Master's program in the lab working on a project related viral capture and concentration using MILs and similar reagents. Sloane has also mentored a junior graduate student and undergraduate in the Moore lab, both of whom will be conducting further research on MIL-based sample preparation. Lastly, Sloane was able to successfully defend her dissertation, which was based largely on work related to MILs. Additionally, Sloane successfully defended her PhD during this period, and one of the undergraduates she was supervising graduated and went on to continue an MS doing this project. Maria Dugan (Student, IA State) attended the Gordon Research Conference on Ionic Liquids in Newry, Maine in August 2024. She presented poster presentations describing the USDA funded work on DESs as nucleic acid extraction media in both the Gordon Research Seminar as well as the Gordon Research Conference. Additionally, Maria presented a poster on the USDA work at the Iowa State University chemistry department open house and was chosen as a finalist to present her work to the chemistry department for the Cotton-Uphaus competition.The project has facilitated extensive professional growth opportunities. Highlights include presentations at prestigious conferences such as the 2024 Gordon Research Seminar (GRS) and Conference (GRC), showcasing our work on sustainable DESs in DNA extraction and qPCR integration. Additionally, I received the Witiak Graduate Fellowship and was named a finalist for the Cotton-Uphaus Award which recognized our commitment to advancing analytical chemistry. The Brehm-Stecher lab (IA State) has leveraged departmental recruitment funds to attract a PhD student for work on this USDA-NIFA project. Ngouana Moffo Auriol Ivane, (MS Food Science and Engineering, Jiangsu University), was awarded the inaugural Iowa State Food Sciences Laboratories TA/RA fellowship. This award provides stipend, tuition, benefits, and fees for a 2-year period, representing additional funding (~$90,000) that we are leveraging to accomplish this project's goals. How have the results been disseminated to communities of interest?These results have been deseminated through numerous scientific forums in the form of peer-reviewed publications, posters, technical talks, and invited seminars. Additionally, the work has been mentioned and referenced in popular press (Boston 25 News: Norovirus:https://www.boston25news.com/news/health/stopping-norovirus-how-umass-amherst-is-working-prevent-spread-dreaded-illness/QOEJ7ZVYHJGAFKNP7GHK2CRFUM/). The work has also been discussed through various visits to other scitneitific institutes and universities by the PIs of the grant and students. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue to evaluate the ability of MILs and DESs to concentrate noroviruses and their RNA, as well as establish and evaluate these reagents for hepatitis A virus. We also plan to develop and optimize anchoring of anti-norovirus aptamers to MILs and evaluate the performance of these liquids in relevant foods for noroviruses.

Impacts
What was accomplished under these goals? In previous years we sought to develop capture and concentration protocols for non-enveloped virus and viral ssRNA using MILs that did not require electrical equipment. To meet this objective, we evaluated the potential of MILs to capture and recover both intact viral particles and purified RNA of bacteriophage MS2, a human norovirus surrogate, from aqueous suspension using a base protocol adapted from a previous study exploring MIL-based capture and concentration of bacterial cells. Through this we established that MILs have potential for capture and recovery of both intact virus and free RNA, as well as for target enrichment. This protocol also requires no centrifugation to separate the MILs from suspension, making this a low-equipment method. We also ran a preliminary experiment to evaluate the performance of a MILs as capture reagents in complex food matrices, and found that MILs can be effective sample preparation reagents in complex suspensions, with performance being significantly impacted by the buffer used to prepare the sample for analysis. More recently, we evaluated the potential of MILs as binding substrate for viral ssRNA extraction. We first adapted our MIL-based capture and recovery protocol for use with eukaryotic human norovirus surrogates, specifically murine norovirus and Tulane virus. We then ran a preliminary experiment comparing MILs to the magnetic silica beads provided in a commercial viral RNA extraction kit, in which we used the same lysis, wash, and elution buffers with both binding substrates. We found that some MIL formulations gave comparable viral RNA recovery to the magnetic silica beads, thereby demonstrating that single-tube viral capture, concentration, and genomic extraction is possible using MILs as a binding substrate. We then developed reagents for chemical lysis, wash, and elution of viral RNA specifically for use with MILs, and identified a simple buffer scheme that gave comparable target recovery to the commercial extraction method, but with reduced time and resources required. This work represents a significant step toward in-field sample processing and detection of foodborne viruses. Our work has focused at developing magnetic DESs that are compatible with biological assays, including PCR and isothermal amplification. Our activities have largely been focused on understanding structural composition aspects of DESs that make them capable of extracting nucleic acids from complex samples and being able to be amplified in downstream analyses. A paper on this work was recently submitted for peer review to Analytica Chimica Acta and is under peer review. Our second goal in this period has been focused at developing new methodologies of preparing mDESs to make them more paramagnetic (able to be captured by permanent magnets) whileachieving extraction of nucleic acids. Compared to MILs, mDESs don't require complicated synthetic methods, but achieving solvents that can withstand leaching of the paragmagnetic metal is more complicated. Our team has achieved methods that will enable more streamlined design of mDESs and we are currently evaluating their efficacy in amplification methods, including qPCR.We have made significant progress in evaluating deep eutectic solvents (DESs) for nucleic acid (NA) extraction and direct integration into quantitative polymerase chain reaction (qPCR) assays. Over one hundred DESs spanning seven classes were screened, assessing their compatibility with qPCR and utility as extraction solvents. Notably, six DESs demonstrated successful DNA amplification, with three achieving efficiencies exceeding 97% for a 121 bp uidA fragment. This study demonstrates, for the first time, DESs' potential to streamline NA analysis workflows by eliminating intermediate purification steps.

Publications

  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2025 Citation: Stoufer S, Kim M, Anderson J, De Silva S, Brehm-Stecher BF, Moore MD. 2025. Evaluating the capacity of magnetic ionic liquids for separation and concentration of non-enveloped viral particles and free viral genomic RNA. Analytical and Bioanalytical Chemistry 417(2):435-445.
  • Type: Peer Reviewed Journal Articles Status: Submitted Year Published: 2025 Citation: Stoufer S, Dugan M, Anderson J, Brehm-Stecher B, Moore MD. 2025. Single-Tube Capture, Concentration, and Genomic Extraction of a Human Norovirus Surrogate Using Magnetic Ionic Liquids. Analytical Chemistry. (In Revision--Minor Revision).
  • Type: Peer Reviewed Journal Articles Status: Under Review Year Published: 2025 Citation: Dugan M, Anderson J. 2025. Exploiting deep eutectic solvents for direct DNA extraction and seamless integration into the quantitative polymerase chain reaction. Analytica Chimica Acta. (Under Review).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Brehm-Stecher, B.F., Moore, M.D., and J.L. Anderson. Magnetic Ionic Liquids for Capture and Concentration of Foodborne Pathogens: Current and Future Applications, Gordon Research Conference on Ionic Liquids as Transformative Materials for Addressing Challenges in Sustainability, Energy and Health, August 4  9, 2024, Grand Summit Hotel at Sunday River, Newry, Maine
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Brehm-Stecher, B.F. Food Safety Applications of Magnetic Liquids, UW-Madison Food Research Institute FRESH Seminar, November 12, 2024, Madison, WI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Grand Summit Hotel, Newry, ME. Exploiting Sustainable Deep Eutectic Solvents for Direct DNA Extraction and Integration into Quantitative Polymerase Chain Reaction. Gordon Research Seminar (GRS). August 4th, 2024
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Grand Summit Hotel, Newry, ME. Exploiting Sustainable Deep Eutectic Solvents for Direct DNA Extraction and Integration into Quantitative Polymerase Chain Reaction. Gordon Research Conference (GRC). August 5th and 6th, 2024
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Stoufer S, Soorneedi A, Kim M, Moore MD. 2024. Sample concentration and processing methods for viruses from foods and the environment prior to detection. Annual Review of Food Science and Technology 15: 455-472.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Stoufer S, Dugan M, Anderson J, Brehm-Stecher B, Moore MD. Development of a Portable Method for Single-Tube Capture, Concentration, and Genomic Extraction of a Human Norovirus Surrogate Using Magnetic Ionic Liquids, Northeast Center to Advance Food Safety Annual Conference and Meeting, January 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Development of a portable method for single-tube capture, concentration, and genomic extraction of non-enveloped virus using magnetic ionic liquids, Pioneer Valley Microbiology Symposium, March 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Development of a portable method for single-tube capture, concentration, and genomic extraction of Tulane virus, a human norovirus surrogate, using magnetic ionic liquids, IAFP Annual Meeting, July 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore MD.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 MD. 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: 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: 2024 Citation: Moore MD. Moore Lab: Applied and Environmental Virology. CNS Rising Stars in Research Innovation  informal meeting with VIP UMass CNS alumni at the invitation of UMass Chancellor Reyes and CNS Dean Fox to discuss Moore lab research. Amherst, MA, USA. 11/16/2024.