The Center for Food Safety Engineeringg 2007 - 2008 Research Report “Collaborating to make our food safer” The mission of the Center for Food Safety Engineering is to develop new knowledge, technologies and systems for detection and prevention of chemical and microbial contamination of foods. Through CFSE, Purdue University positions itself as a national leader in multi- disciplinary food safety research. Our multi- disciplinary approach, including a strong engineering component, makes Purdue University truly unique. 2007-2008 Research Report 2 Welcome from the Director • Message from Richard Linton, Center Director 2 Message from USDA • Message from our Partnership with USDA-ARS 3 Multipathogen screening using immunomicroarray • Arun Bhunia 4 Optical biosensors for food pathogen detection • Arun Bhunia 5 Optical forward scattering for bacterial colony differentiation and identifi cation • Arun Bhunia, E. Daniel. Hirleman, J. Paul Robinson 6 Immunocapture real-time PCR to detect mycotoxigenic mold spores in grains • Maribeth A. Cousin, Charles P. Woloshuk 7 Detection of foodborne pathogens via an integrated spectroscopy and biosensor-based approach • Joseph Irudayaraj, Lisa Mauer, Chitrita DebRoy, Pina Fratamico 8 Nanoparticle-based DNA-multiplexed probes for pathogen detection using confocal raman microscopy • Joseph Irudayaraj 9 Engineering of biosystems for the detection of Listeria monocytogenes in foods • Michael R. Ladisch, Rashid Bashir, Arun Bhunia, J. Paul Robinson 10 Spotlight on USDA-ARS Scientists 12 Rapid, quantitative, and reusable immunosensors for bacteria detection on a microfl uidic platform • Chang Lu, Arun Bhunia, Zhongyang Cheng 13 A method for capture and detection of Escherichia coli O157:H7 using polymer-immobilized phage • Mark Morgan, Bruce Applegate 14 Continuous monitoring of chemical agents in aqueous media using bioreporter-based sensors • David Nivens, Michael Franklin, Carlos Corvalan 15 Field-ready biosensors for high throughput and multiplexed detection of foodborne pathogens • Kinam Park, James Leary, Arthur Aronson 16 Portable biosensor for rapid and ultra-sensitive identifi cation of organophoshorous foodborne contaminants • Lia Stanciu, Silvana Andreescu 17 Scientifi c Publications and Presentations 20 Center Staff Visit us @ www.cfse.purdue.edu Welcome from the Director The year 2008 marks the eighth anniversary of the Center for Food Safety Engineering (CFSE) at Purdue University. Our partnership with the United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Eastern Regional Research Center continues to create signifi cant research and outreach impacts. This year, the CFSE team published 31 peer-reviewed research publications and presented 18 talks at national science meetings. CFSE scientists also have been forming partnerships with international countries. Scientifi c presentations and collaboration- seeking visits were made this year to Brazil, China, Hong Kong, and Scotland. Our trip to Jiao-Tong University in Shanghai, China was especially important in fostering a new international collaboration between USDA-ARS and Jiao-Tong University. This year, our research program underwent a USDA-ARS Offi ce of Scientifi c Quality Review Dr. Richard H. LintoLintonn (OSQR), and the future direction of our research program was identifi ed. A research priority for DirectorDirector ofof the Center forfor the next few years is development of technological platforms that improve microbial and chemical Food SafetySafety EngineeringEngineering hazard detection. These new technologies will advance detection of bacterial pathogens including 765.494.6481765 494 6481 Listeria monocytogenes, Escherichia coli O157:H7, Campylobacter spp., and Salmonella spp. and [email protected] of chemical hazards that may present food safety and food defense concerns. Our detection-based technologies will build upon prior success with optical biosensors, cell- based biosensors, bio-chips (lab-on-a-chip), microarrays, infrared spectroscopy (including Fourier transform infrared FTIR), enzyme linked immunosorbant assays, polymerase chain reactions, impedance-based microbiology, scanning microscopy, confocal raman microscopy, bioluminescence, DNA/RNA probes, and bioreporter-based chemical sensors. The exciting BActeria Rapid Detection using Optical scattering Technology (BARDOT) system is being evaluated by industry and regulatory agencies, and we hope that it will be available for wide-scale use sometime next year. I continue to be impressed with the collaborative research efforts of Purdue University and USDA- ARS scientists and feel privileged to serve as director of the center. If you are interested in learning more about CFSE, please visit our Web site at www.cfse.purdue.edu or contact me directly. Message from USDA As the collaboration between the CFSE at Purdue University and the USDA-ARS Eastern Regional Research Center continues, I am grateful to witness the continual growth, maturation, and research impact of this partnership. We are pleased to learn that the ARS-Purdue team has successfully completed the USDA-ARS OSQR process with outstanding ratings. This result signals that the ARS-Purdue project is now considered to be an integral part of USDA-ARS research efforts. Together, we have received increased recognition as an important contributor to the technological advancement of pathogen detection in food—evidenced by the fact that the team received invitations from the International Workshop on Rapid Methods and Automation in Microbiology to conduct half-day symposiums on molecular methodologies in August 2006, June 2007, and June 2008. The team received extraordinary praise from Dr. Daniel Y. C. Fung, Professor of Food Microbiology, Kansas State University, for invaluable contributions to the workshop, and Dr. Shu-I Tu he extended an invitation for our continuing involvement in 2009. In May 2008, our team went to Supervisory Research Shanghai, China to attend the fi rst annual meeting of the Joint United States-China Food Safety Chemist USDA-ARS, Center, an international collaboration established between the ARS and Jiao-Tong University in Eastern Regional Shanghai. This collaboration is part of the cooperative research activities between the USDA and Research Center the Ministry of Science and Technology, China (MOST). With this collaboration in place, I believe 215.233.6466 that our ARS-Purdue team will become an important international research enterprise in the near [email protected] future. 2 Center for Food Safety Engineering Bhunia Multipathogen screening using immunomicroarray Investigator: Arun Bhunia (Department of Food Science) Project Rationale Project Objectives Antibody-based methods for detecting pathogenic foodborne • Develop a microarray assay in 96-well plate and bacteria are used widely and are regarded as rapid and effi cient. glass slide using sandwich immunoassay for Application of conventional antibody-based assays and further three pathogens: Salmonella spp., Escherichia adaptation in modern biosensor tools show promise for rapid coli O157:H7, and Listeria monocytogenes detection. Most assays are developed for detection of a single • Optimize growth and enrichment of these pathogens target pathogen or toxin. As a result, these methods can be (healthy or stressed) spiked in model food samples in costly when testing for multiple pathogens from a single product a selective enrichment broth for use with microarray. because separate assay methods are required. In addition, a • Evaluate performance of the Pathogen large laboratory space is required to perform separate tests for Enrichment Device (PED). each target pathogen because separate enrichment media and procedures are needed for each pathogen detection method. Project Highlights The development of a single test capable of detecting multiple For simultaneous growth and detection of Salmonella spp., pathogens enriched in a single enrichment media would reduce E. coli O157:H7, and L. monocytogenes, we formulated a costs and yield quick results. This technology would also benefi t selective enrichment broth SEL (Salmonella, E. coli O157:H7, regulatory agencies in the evaluation of food products for key and Listeria) and found it to be suitable for enrichment of all food pathogens. these target pathogens in spiked ready-to-eat deli turkey and salami. In addition, when we evaluated the performance of SEL In the past two decades, several rapid detection methods, for its ability to detect pathogens by immunoassay, we found such as antibody-based, nucleic acid-based, and biochemical- that target pathogens grown in SEL gave a stronger response based methods, were developed. Even though these methods than when they were grown on other media. The global have shortened analysis times, it still takes time for selective protein expression profi les of the three pathogens in SEL were enrichment of samples before conventional rapid detection signifi cantly greater than their respective selective enrichment methods can be employed. Antibody-based methods, such as broths. Proteomic analysis further revealed the presence of a enzyme-immunoassay, require a minimum of 106 CFU/ml for cell unique protein in bacteria when grown on SEL. detection. To achieve that cell level, it is important to use proper enrichment media for detection of foodborne pathogens. Cell injury or stress encountered during food processing may affect cell numbers. The presence of selective agents in media can delay growth and recovery of stressed or injured cells.