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In the U.S. alone, there are an estimated 47.8 million illnesses, over 127,000 hospitalizations and 3,000-plus deaths attributed to foodborne illness each year. Traditional means for microbial detection can no longer match the pace of today’s food processing and global distribution networks. Emerging rapid sensor and detection platforms can provide the timely and actionable information needed to lessen the human and economic burdens levied by foodborne disease.

This symposium will feature presentations on emerging optical, nanotechnological, spectroscopic and electrochemical technologies for pathogen detection, including label-free and high-throughput methods. Novel ligands for pathogen capture will also be examined. Benefits and challenges of these new methods and their comparison with existing techniques will be discussed. Label-free approaches and the advantages of novel bioaffinity ligands will be highlighted.In the U.S. alone, there are an estimated 47.8 million illnesses, over 127,000 hospitalizations and 3,000-plus deaths attributed to foodborne illness each year. Traditional means for microbial detection can no longer match the pace of today’s food processing and global distribution networks. Emerging rapid sensor and detection platforms can provide the timely and actionable information needed to lessen the human and economic burdens levied by foodborne disease.

Final Agenda

Monday, June 26

(Shared Afternoon Plenary Session with Biodetection Technologies: Pathogen Detection Track)

Agro-Defense, Toxins & Allergens

1:55 pm Chairperson’s Opening Remarks

Willy Valdivia-Granda, Ph.D., CEO, Orion Integrated Biosciences

2:00 Agro-Defense - A Holistic, All of Enterprise Approach

Tammy R. Beckham, D.V.M., Ph.D., Dean, Professor, College of Veterinary Medicine, Kansas State University

The ability to protect our agricultural industries, food supply, and public health sectors from natural introductions of biological agents, agro-terror threats, and emerging and re-emerging diseases is heavily dependent on an organized, strategic, and well-funded approach. This approach should institutionalize the “One Health” concept, be highly collaborative in nature, leverage all available resources and encompass an international, global health component. The One Health concept must be understood, adopted and become part of the fabric of the way in which we approach biodefense.

2:30 xMAP® Multiplex Detection: Getting Beyond Detection to Include Built-In Confirmation, Characterization, and the Ability to Distinguish between Unanticipated Homologous Analytes

Eric Garber, Ph.D., Division of Bioanalytical Chemistry, Office of Regulatory Science, Center for Food Safety and Applied Nutrition, FDA

By using antibody-based multiplex methods (e.g., xMAP®), it is possible to generate antigenic profiles along with other second-order forms of data processing. These results can provide built-in confirmation, recognition, and characterization of unique features, as well as the detection of novel unexpected analytes. Such approaches have been applied to the detection of toxins and recently a commercial assay was developed for the detection of food allergens to meet the complexity of a growing global marketplace and an increase in the apparent prevalence of food allergies.

Screening, Capture & Biosurveillance

3:00 Development of 11-Plex MOL-PCR Assay for the Rapid Screening of Samples for Shiga Toxin-Producing Escherichia coli

Alina Deshpande, Ph.D., Group Leader, Biosecurity and Public Health, Bioscience Division, Los Alamos National Laboratory

Shiga toxin-producing Escherichia coli (STEC) strains are a serious public health threat with about half of the STEC related foodborne illnesses attributable to contaminated beef. Los Alamos National Laboratory (LANL) has developed an assay that can screen samples for several important STEC-associated serogroups (O26,O45,O103,O104,O111,O121,O145,O157) and three major virulence factors(eae,stx1,stx2) in a rapid and multiplexed format using the Multiplex oligonucleotide ligation-PCR (MOL-PCR)assay chemistry. This presentation describes the development and testing of the 11-plex assay that could serve as the backbone for high-throughput screening systems for both routine screening and outbreak investigation.

3:30 Refreshment Break in the Exhibit Hall with Poster Viewing

4:15 Broad-Based Detection and Classification of Pathogens Using Antimicrobial Peptides for Pathogen Capture

Chris Taitt, Ph.D., Research Biologist, United States Naval Research Laboratory

Many current biodetectors are designed either to detect a limited number of known pathogens or to determine whether something biological is present, with little further characterization. Here, we use broad-based, semi-selective binding molecules - antimicrobial peptides - to detect a broad variety of microbes, with classification/discrimination based on binding patterns. We have recently initiated efforts to use the unique structure/function relationship of these same peptides to achieve “reagentless” sensing.

4:45 Genomic-Based Characterization and Biosurveillance of Known and Unknown Microorganisms of Biodefense Relevance

Willy Valdivia-Granda, Ph.D., CEO, Orion Integrated Biosciences

Next generation sequencing technologies offer an unparalleled opportunity to detect and characterize microbes of public health and agrodefense concern. However, the simultaneous analysis of hundreds of genomic and metagenomic datafiles generated by different sequencing platforms remains complex. Here we present a ubiquitous approach for microbial identification that generates molecular taxonomic profiles of known and unknown microbes contextualized with geospatial attributes for attribution, bioforensics, biosurveillance and risk assessment. The implications of our system for deterrence and biodefense will be discussed.

5:15 Welcome Reception in the Exhibit Hall with Poster Viewing

6:15 End of Day

Tuesday, June 27

8:00 am Morning Coffee

8:55 Chairperson’s Opening Remarks

Byron Brehm-Stecher, Ph.D., Associate Professor, Rapid Microbial Detection and Control Laboratory, Iowa State University

9:00 KEYNOTE PRESENTATION: Food Safety Microbiology in the Metagenomics Era

John_BesserJohn Besser, Ph.D., Deputy Chief of the Enteric Diseases, Centers for Disease Control & Prevention

Metagenomics, once the sole realm of research labs, will soon become practical for many food safety activities. Together, these advancements will change how we monitor foods and food production environments, detect and investigate outbreaks, and will fundamentally change our understanding of foodborne disease. Current sequencing-based activities for CDC’s food safety programs will be described, along with a discussion of exciting possible applications in the not-too-distant future.

9:30 FEATURED PRESENTATION: Understanding Lower Limits of Detection: Promising You Can Detect a Single Bacterial Cell in Food Samples Is One You Cannot Deliver

Robert_BuchananBob Buchanan, Ph.D., Professor & Director, Center for Food Safety & Security Systems, University of Maryland

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

Optical & Genetic Approaches for Detection & Characterization of Pathogens

10:45 Rapid Detection of Enterobacteriaceae as Indicator for Pathogen Testing Using a Light Scattering Sensor

Arun Bhunia, Ph.D., Professor of Food Microbiology, Department of Food Science, Department of Comparative Pathobiology, Purdue University

The Enterobacteriaceae (EB) family and its most known members, coliforms, are used as “indicators” of hygiene monitoring, sanitization practices and process verification of food products. Colony scatter images were analyzed using image classifier and results show BARDOT can potentially be used for differentiation and identification of members of EB family and the coliforms for use in food process verification, hygiene monitoring, or food safety. (Contributing authors: Marcela Martinez and Atul K. Singh)

11:15 Genetic-Based Methods for Characterization, Detection, and Typing of E. coli

Pina M. Fratamico, Ph.D., Research Microbiologist, Research Leader, Agricultural Research Service, Eastern Regional Research Center, USDA

Developments in omic- and genetic-based technologies are enhancing the ability to detect, identify, and characterize E. coli. Whole genome sequencing (WGS) is providing a powerful and expanding range of information to identify targets for the development of rapid and specific detection and identification systems for E. coli and other bacteria and is being implemented for source tracking and as part of routine surveillance systems. It is expected that further developments in WGS and other genomic and molecular technologies will continue to contribute to the development of improved detection and identification systems and to a greater understanding of the pathogenesis of E. coli and ultimately provide better resources for improving public health.

11:45 Sponsored Presentation (Opportunity Available)

12:15 pm Lunch on Your Own

Methods Overview

1:40 Chairperson’s Remarks

Arun Bhunia, Ph.D., Professor of Food Microbiology, Department of Food Science, Department of Comparative Pathobiology, Purdue University

1:45 Advances in Pre-Analytical Sample Preparation: Unclogging the Bottleneck from Sample to Answer

Byron Brehm-Stecher, Ph.D., Associate Professor, Rapid Microbial Detection and Control Laboratory, Iowa State University

Successful detection of pathogens in foods involves the seamless integration of three interdependent steps: 1) statistically validated sampling, 2) pre-analytical sample preparation and 3) detection. Weak links in any of these three steps will propagate through the system and degrade the end result. In a worst-case scenario, this could lead to false-negative results. “Upstream” methods for sampling have long been established, and the past decade has seen a revolution in development of the hardware and reagents needed for truly rapid detection of pathogens.

2:05 Nucleic Acid Aptamers as Bioaffinity Ligands for Detection of Human Norovirus

Lee-Ann Jaykus, Ph.D., Professor, Department of Food, Bioprocessing & Nutrition Sciences, North Carolina State University

Single-stranded DNA molecules (nucleic acid aptamers) that naturally fold into complex three-dimensional shapes with target binding affinity to various foodborne pathogens have been identified by several groups. Those with binding specificity to human norovirus, the leading cause of food-associated disease, can be used for pre-analytical sample processing, detection, infectivity determination, study of virus structure, and potential therapeutics. In this presentation, we will discuss all these possible functions in the context of norovirus aptamer candidates produced using a variety of selection approaches and evaluated for use in pristine and complex sample matrices.

2:25 Development and Application of NGS toward Detection of Norovirus at Low Copy Number as a Single or Mixture of Viral Species Extracted from Food/Celery

Zhihui Yang, Ph.D., Molecular Virology Team, Division of Molecular Biology, Office of Applied Research and Safety Assessment, U.S. Food and Drug Administration

Next generation sequencing holds promise as a single application for both detection and sequence identification of foodborne viruses. However, technical challenges remain due to anticipated low quantities or mixtures of viruses in contaminated food. In our current study, with a focus on data analysis with various bioinformatics tools, we present our approach toward the amplification-independent detection of norovirus at low copy number individually or within a mixture of two virus species/strains extracted from celery.

2:55 Bacteriophage Engineering for the Onsite Detection of Pathogens and Indicators

Emma Farquharson, Ph.D. Student, Nugen Lab, Cornell University

Bacteriophages continue to offer new tools for advancing food safety. By modifying phages to be amenable for deployable, low-cost, and rapid assays - this opens the door for significant improvements to be made to current portable methods. This allows phage-based assays to be used in resource-limited areas such as those encountered in food and agriculture.

3:15 Nucleic Acid Biosensors for the Detection of Pathogens

Lingwen Zeng, Ph.D., Researcher, Institute of Environmental and Food Safety, Wuhan Academy of Agricultural Science and Technology, China

Pathogens are recognized as widespread and toxic contaminants that can cause deleterious effects on food safety and human health. In our lab, we focus on developing nucleic acid biosensors for pathogens detection using aptamers as the molecular recognition elements. Employing colorimetric analysis, lateral flow strip biosensor, and fluorescence detector as the sensing platforms, our proposed biosensors would be promising strategies for pathogens detection which offer prominent advantages of improved sensitivity and convenience.

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing

4:15 Label-Free Protein Detection Based on the Heat-Transfer Method – A Case Study with the Peanut Allergen Ara h1 and Aptamer-Based Synthetic Receptors

Marloes Peeters, Ph.D., Assistant Professor, Chemistry & Environmental Science, Manchester Metropolitan University, United Kingdom

Aptamers are an emerging class of molecules which, due to the development of the systematic evolution of ligands by exponential enrichment (SELEX) process, can recognize virtually every target ranging from ions, to proteins, and even whole cells. We will present the heat-transfer method (HTM) as an interesting alternative since this offers detection in a fast and low-cost manner and has the possibility of performing experiments with a fully integrated device. This concept has been demonstrated for a variety of applications including DNA mutation analysis and screening of cancer cells, but the concept of using this for food safety (allergen) detection is novel.

4:45 New Advanced Colorimetric Assay Performed during the Enrichment Process for the Detection of Foodborne Pathogens

Bruce Applegate, Ph.D., Professor, Center for Food Safety Engineering, Purdue University

Methodology for the detection of foodborne pathogens requires an enrichment step which is the time sink in most protocols. A colorimetric method which exploits this step for detection has been developed for E. coli O157:H7. The method also allows for the selective recovery of the pathogen if present and can be developed for other organisms as well.

5:15 End of Rapid Detection for Food Safety Track and Workshop Registration

5:30 Workshop 1: Rapid Sample Preparation for Pathogen Detection*

Instructor: Dave Alburty, CEO, InnovaPrep LLC

*Separate registration required for workshop

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