Agent Behavior

The National Center for Food Protection and Defense is a multi-disciplinary and action-oriented research consortium. Collaborations have been established at over 40 academic and private sector organizations.

The goal of the Agent Behavior theme is to develop innovative detection, decontamination and inactivation technologies using research on the fundamental behaviors of select chemical or biological agents in food.

Agent Behavior Publications

Agent Behavior Theme Leaders

Peter Varelis, Fonterra
Eric Johnson, University of Wisconsin-Madison

Current Projects

Castor beans, the source of ricin

Application of Surface Enhanced Raman Spectroscopy for Detection of Chemical and Biological Terror Agents in Food Matrices
Ted Labuza, University of Minnesota
Sponsor: DHS/NCFPD

The aim for this project is to develop and validate Surface Enhanced Raman Spectroscopy (SERS) assays for detection of ricin and anthrax spores in liquid foods within 20 minutes. Initially, SERS had been used successfully to assess its capabilities of detecting and differentiating bacterial spores from several species of Bacillus.

Subsequently an aptamer-based SERS assay was validated for ricin in liquid food detection at 0.5 ppm in milk and 0.1 ppm in orange juice.

To address the B. anthracis spore challenge, immunomagnetic separation (IMS) was used on milk and orange juice. Several antibodies were tested for IMS and the procedure optimized SERS was subsequently used to measure the dipicolinic acid (DPA) in the IMS captured spores so that in combination they represent a rapid method of B. anthracis spores. Hand held SERS instruments should result in strong potential for transition of this methodology.

Enrichment Media

Srinand Sreevatsan, University of Minnesota
Sponsor: DHS

The Enrichment Media project provides research to develop new pathogen enrichment protocols that are critical to meeting the demands of the Ag Defense agriculture screening tools program. Newly developed approaches for Francisella tularensis and Yersinia pestis will utilize traditional empirical methods, more focused methods based on comparative genomics, physical selection using capture tools, and selective removal of likely competitors. Francisella tularensis and Yersinia pestis can grow over night in a pure culture but when in a matrix of food or consumable liquids and other bacteria, but these bacterial threat agents can be hard to identify. Proposed enrichment protocols include: a) to use physical means to either isolate Francisella tularensis or Yersinia pestis from a mixture of competitor microbes or to selectively remove the competitor microbe, and b) to selectively remove Francisella tularensis or Yersinia pestis cells from a mixture using aptamers or specific antibodies. To date, results utilizing spent media are promising. These fundamental findings have the potential of method modification and development not only for the microorganisms at test but for other pathogens in food.

Modification of Sample Preparation to Differentiate Live, Injured and Dead Bacteria in Polymerase Chain Reaction Assay
Teshome Yehualaeshet, Tuskegee University
Sponsor: DHS/NCFPD

Initially, the inactivation of the test microorganisms had to be characterized and validated under traditional standardized conditions.

Thermal injury and recovery of Yersinia was studied: a) to predict the kinetics of thermal death/ injury and recovery of Yersinia in milk under different temperature profiles; b) to estimate D-/and Z-value of Yersinia in milk and c) to investigate the impact of media supplement in the recovery process of Yersinia after temperature stresses for the evaluation of cell injury. The use of flow cytometry and standard serial dilution was used to determine the injury and recovery of Yersinia after thermal stresses.

Modification of PCR sample preparation was used to delineate live, injured and dead bacteria. A collection of DNA-intercalating chemicals was selected and screened to determine which are highly selective in penetrating only into ‘dead’ bacteria to subsequently block the amplification of the targeted gene. These DNA-intecalating chemicals were also assessed to examine if the chemical works similarly on DNA extracted from both culture and food samples. Expansion of these findings could greatly influence the standard assessments of efficacy of microbial control processes for food systems.

Modular and Expandable Detection Platform for Current and Potential Food Toxins and Adulterants
Lawrence Wackett, University of Minnesota
Sponsor: DHS/NCFPD

The overall objective of the present work is to develop a general platform that can be used for detecting thousands of chemical toxicants. The specific aim is to obtain enzymes and develop methodologies for several chemical threats in foods. The compounds of specific interest are ricinine (a surrogate for ricin), cyanide, acrylamide (a neurotoxin), azetidine-2-carboxylate (the toxic agent in Lily of the Valley), nicotine, semicarbazide, caffeine, and other toxicants.

Specific enzymes have been isolated for the initial agent under test and the overall refinement of the methodology is underway. It is anticipated that the procedure under study that successfully detects specific agents in food will be readily transitioned to commercial kits as was Wackett and Sadowsky's earlier successful method for melamine.

A smart temperature-sensitive liquid microlens using NIPAAm hydrogel.

From the following article: Adaptive liquid microlenses activated by stimuli-responsive hydrogels. Liang Dong, Abhishek K. Agarwal, David J. Beebe and Hongrui Jiang, Nature 442, 551-554 (3 August 2006) doi:10.1038/nature05024

Novel Methods for Detection of Clostridium botulinum and Botulinum Neurotoxin in Complex Food Matrices
Eric Johnson, University of Wisconsin-Madison
Sponsor: DHS/NCFPD

The aims for this project are to develop a BoNT (Botulinum neurotoxin) detection system, specifically focusing on sample preparation to reduce time, effort and loss as compared to current detection techniques. A vertical IFAST colorimetric immunoassay system has been developed for detection of botulinum neurotoxin type A. This allows for easier sample handling and testing larger volumes. This has involved special synthesis of the peptides crosslinker for the hydrogel. Further, two fabrication systems have been used to fabricate the crosslinked hydrogel structures. Continued optimization and expansion to different serotypes is planned before transition to possible commerical kits.

On-site Detection of Microbial Pathogens and Chemical Toxic Agents using Novel Sensors Based on Quantum Dot Modified Molecular Imprinted Polymers
Keith Warriner, University of Guelph
Sponsor: DHS/NCFPD

The overall aim of the study is to develop imprinted polymer based sensors for detection of microbes and chemical select agents using the quenching of the fluorescence of quantum dots as the transduction strategy to report the binding events. As a proof of concept Tobacco Mosiac Virus (TMV) is used as a surrogate for a human pathogen. The process first requires optimization of extraction and assay conditions for the detection of the viruses captured on the beads. Once this has been accomplished the validation of sensor performance will be conducted and the tests will be expanded to Pepper Mosiac Virus. Application of this to agriculturally significant plant and mammalian viruses as well as human viruses will be the next transition.

Validation of Agent Inactivation and Isolation from Food Systems
Alvin Lee, Institute for Food Safety and Health, Illinois Institute of Technology
Sponsor: DHS/NCFPD

This project will involve integration of projects funded by NCFPD so that validation of developed protocols by the various research groups could be brought together and validated by applying those findings in large scale situations. This will be accomplished in a secure high-level biocontainment facility at IFSH that houses pilot or full scale food processing equipment. The project will aim to integrate previous laboratory based research outcomes with scale-up activities using pilot or full scale food processing equipment and with the use of threat agents. Such activities will validate research outcomes and provide the food industry with validated protocols for the inactivation or removal of threat agents from processing equipments.

Completed Projects

For more information on completed projects, contact us through our inquiry form.

Bioluminescent Bacteria as Biological Sensors for Toxic Agents in Food
Vangie Alocilja, Michigan State University

Bioluminescent Imaging for High Throughput Screening for Bacterial Pathogens and Toxins
Mansel Griffiths, University of Guelph

Biosensors for Detection of Chemical Toxins
Paul Takhistov, Rutgers University

Botulinum Neurotoxin Sensing Technologies
Eric Johnson, University of Wisconsin-Madison

Capture and Detection of Botulinum Neurotoxin (BoNT) in Complex Food Matrices using Novel Biosensor Platforms
Eric A. Johnson, University of Wisconsin-Madison

Chemical Inactivation of Protein Toxins on Food-Contact Surfaces
Jack Cappozzo and Lauren Jackson, National Center for Food Safety and Technology (NCFST), Illinois Institute of Technology

Concentrating Bacterial Spores from Milk and Juices using Dielectrophoresis-Based Microfluidic Capture Systems
Suresh Pillai, Texas A&M University
Ali Beskok, Old Dominion University

Containment and Remediation System
Susan Harlander, BT Safety LLC

Detection and Removal of Select Agents from Complex Food Matrices: Phase II
Lee-Ann Jaykus, North Carolina State University

Determination of Y. pseudotuberculosis Survival in Milk under Temperature Stress
Teshome Yehualaeshet, Tuskegee University

Development of a Multiplex Bio-barcode DNA Biosensor for Bacillus anthracis Detection Without PCR Amplification
Vangie Alocilja, Michigan State University

Development of biologically modified electrically-active magnetic nanoparticles (nano-BEAMs) for direct capture and concentration of Bacillus anthracis spores and cells in various food matrices
Vangie Alocilja, Michigan State University

Development of Nontoxigenic Clostridium botulinum Toxins and Strains for Food Defense Studies
Eric Johnson, University of Wisconsin-Madison

Electrochemical Biosensors for B. anthracis
Vangie Alocilja, Michigan State University

Evaluation of Methods for Decontamination of Food Processing Equipment and Facilities Deliberately Contaminated with Bacillus Spores
Martin Cole, National Center for Food Safety and Technology–Illinois Institute of Technology
Peter Slade, National Center for Food Safety and Technology–Illinois Institute of Technology

Extraction of Bacillus anthracis Spores and Ricin from Liquid Foods and its Potential Utilization for Inline Detection
Ted Labuza, University of Minnesota
Francisco Diez-Gonzalez, University of Minnesota

Extraction, Concentration, and Detection of Toxins in Solid Food Systems using Molecular Imprinted Polymer Films
Keith Warriner, University of Guelph

Extraction and Concentration of Chemical (Tetramethylenedisulfotetramine) and biological (ricin) toxins using Molecular Imprinted Electrospun Fibers Coupled with Detection at Aptamer Modified Polymer Electrodes
Subrayal Reddy, University of Surrey (UK)
Keith Warriner, University of Guelph

Fate of Toxins in At-Risk Foods
Ramona Parra, New Mexico State University

FASTMAN Integrated Device for Detection of Select Agents
Vivek Kapur, University of Minnesota/ANDX

Heat Inactivation Kinetics of Spores in Liquid Milk
Ted Labuza, University of Minnesota Francisco Diez-Gonzalez, University of Minnesota

Hydrophobic Extraction of B. anthracis Spores from Liquid Foods
Francisco Diez-Gonzalez, University of Minnesota
Ted Labuza, University of Minnesota

Identification and Characterization of Temperature-Dependent Virulent Genes in Y. pestis, Y. pseudotuberculosis and Y. enterocolitica
Teshome Yehualaeshet, Tuskegee University

Impact of Select Agents on the Coagulation Properties of Milk Proteins
Salam Ibrahim, North Carolina A&T State University

Integrating Microbial Target Capture with Flow Field Fractionation for Lab on a Chip Device for Food Matrices
Suresh Pillai, Texas A&M University

Inactivation of Microbial Agents with High Concentrations of Industrial Sanitizers
Katie Swanson, Ecolab Inc.
Bruce Cords, Ecolab Inc.

Melamine-Cyanuric Acid Detection System for Purposely Adulterated Foods
Lawrence Wackett, University of Minnesota

A Microfluidic Approach for Separation and Concentration of Bacterial Spores from Milk and Juices
Suresh Pillai, Texas A&M University
Ali Beskok, Old Dominion University

Plasma Technology to Decontaminate Surfaces
Amy Wong, University of Wisconsin–Madison
Ferencz Denes, University of Wisconsin–Madison

Proof-of-Concept Study for the Development of a Cell-Free GPCR-Based Biosensor for Rapid Non-Specific Detection of Chemical and Biological Toxic Agents
Evangelyn Alocilja, Michigan State University

Protein Complementation Based Functional Detection Assay for Staphylococcal Enterotoxin B
Jody Proescher, Johns Hopkins University

Rapid Testing for Botulinum Toxin using Egg Yolk Antibodies
Mark Cook, University of Wisconsin–Madison

A Simple and Rapid Method for the Simultaneous Purification and Concentration of Nucleic Acids and Proteins from Food Matrices Simultaneous Detection of Multiple Food Safety Threat Agents Using Multiplex PCR and PCR-Based Microarray Approaches
Charles Young, The Johns Hopkins University Applied Physics Laboratory
Woubit Salah Abdela, Tuskegee University School of Veterinary Medicine

A Systematic Approach for the Detection of Bioterrorism Agents in Complex Sample Matrices
Lee-Ann Jaykus, North Carolina State University

Stability of BoNT in Food Matrices
Eric A. Johnson, University of Wisconsin-Madison

Stability of Ricin and Abrin to Conventional Food Processing Operations and Investigations on Food Contact Surface Decontamination Methods
Peter Varelis, National Center for Food Safety and Technology–Illinois Institute of Technology

Thermal and Chemical Inactivation of Select Agent Toxins in Liquid Foods
Ted Labuza, University of Minnesota
Francisco Diez-Gonzalez, University of Minnesota

Toxin/Pathogen Inactivation and Disposal of Intentionally Contaminated Foods
Craig Benson, University of Wisconsin–Madison

Use of Commercial Household Sanitizers to Inactivate Spores
Michael Davidson, University of Tennessee–Knoxville

Validation of Agent Inactivation and Isolation from Food Systems
Alvin Lee, National Center for Food Safety and Technology

Validation of Methods for Decontamination of Food Processing Equipment and Facilities Deliberately Contaminated with Bacillus Spores
Alvin Lee and Robin Kalinowski, National Center for Food Safety and Technology (NCFST), Illinois Institute of Technology