CENTER FOR FOODSAFETY ENGINEERING

Imagine a world where we can detect pathogenic bacteria in a food system within hours rather than days…

…The Center for Food Safety Engineering at Purdue University is developing technologies to do just that.

2014NEWSLETTER

NOTE FROM THE DIRECTORThe Center for Food Safety Engineering (CFSE) at Purdue University was established in 2000 as a partnership with the United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Eastern Regional Research Center (ERRC). The mission of the CFSE is to develop new knowledge, technologies, and systems for detection and prevention of chemical and microbial contamination of foods while training the next generation of food safety scientists and engineers. Our researchers are developing systems that use advanced engineering principles coupled with microbiological techniques. These systems include:

• C3D: Effective food sampling protocols and filtration techniques to maximize microbial cell separation and concentration in an automated instrument.

• Biochip: Biochip systems, using immunobiology and electrochemistry, for detecting viable cells of Salmonella enterica serovars, Shiga toxin-producing Escherichia coli (STEC), and Listeria monocytogenes in food.

• BARDOT: The Bacterial Rapid Detection using Optical scattering Technology (BARDOT) system for microcolony detection and identification of bacteria, including pathogenic L. monocytogenes, select Salmonella serovars, and STEC.

• Phage: A one-step method using bacteriophages carrying reporter genes for detecting E. coli O157:H7, STEC, and other foodborne pathogenic bacteria directly in the enrichment bag.

• Raman Sensor: A Raman biosensing platform for detecting single pathogenic cells.• And many others, such as next-generation metagenomic sequencing methods, immunocytochemistry techniques, and

immunobiosensors.

There were many highlights and banner accomplishments during 2013-2014. The CFSE participated in the 2013 Annual Meeting of the MOST-USDA Joint Research Center for Food Safety in Shanghai, China, with additional laboratory visits and presentations at Shanghai Xiao Tong University and in Nanjing, China. CFSE members circled the globe, participating in important food safety conferences and activities from our home base to China, Afghanistan, Ireland, and beyond.

This newsletter contains only an overview of the many activities and accomplishments of the CFSE. If you are interested in learning more about the CFSE, please visit our Web site at www.cfse.purdue.edu or contact me directly. Together, we can work to ensure the safety of the global food supply.

Director Dr. Lisa Mauer Professor, Department of Food Science

Annual Highlights ..............................................3Research Project Overviews ............................. 5Scientific Team Recognitions ........................... 9Scientific Publications ....................................10

TABLE OF CONTENTS

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As the collaboration between the Center for Food Safety Engineering (CFSE) at Purdue University and the USDA, Agricultural Research Service (ARS) Eastern Regional Research Center (ERRC) continues, I am pleased to witness the growth, maturation and impact of this partnership. The partnership is considered an important part of USDA-ARS efforts in food safety research, addressing high priority ARS research goals focused on foodborne pathogens and pathogen detection technologies.

The outstanding research and the growth of the CFSE technologies were evident by the quality and variety of the presentations made by CFSE scientists at the ERRC-CFSE Annual Meeting held at the ERRC in January of this year. In addition to this annual meeting, USDA-ARS and CFSE scientists were invited to travel to China to participate in scientific meetings of two collaborative research centers established between the USDA and The China Ministry of Science and Technology (MOST): The USDA-MOST Virtual Food Safety Research Center at Shanghai Jiao Tong University and The China Joint Research Center for Food Quality and Safety Control at Nanjing Agricultural University; providing CFSE scientists an international forum in which to present their research accomplishments.

As the ARS principal investigator and collaborative participant, I look forward to the continued growth and maturation of the CFSE technologies and the ERRC-CFSE partnership.

Dr. George C. Paoli USDA, ARS Principal Investigator

Device speeds concentration step in food-pathogen detection Oct. 14, 2013 | www.phys.org/news

(Emil Venere) -- Researchers have developed a system that concentrates foodborne Salmonella and other pathogens faster than conventional methods by using hollow thread-like fibers that filter out the cells, representing a potential new tool for speedier detection. The machine, called a continuous cell concentration device, could make it possible to routinely analyze food or water samples to screen for pathogens within a single work shift at food processing plants. “This approach begins to address the critical need for the food industry for detecting food pathogens within six hours or less,” said Michael Ladisch, a distinguished professor of agricultural and biological engineering at Purdue University. “Ideally, you want to detect foodborne pathogens in one work shift, from start to finish, which means extracting the sample, concentrating the cells and detection.” Findings are detailed in a research paper appearing in the November issue of the journal Applied and Environmental Microbiology.

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IN THE NEWS

Spring FestScientists in the Center for Food Safety Engineering at Purdue University are teaching the next generation about microorganisms in food. CFSE faculty and students showed youngsters how to plate spinach and tests for microorganisms during Spring Fest, an annual event that draws over 30,000 people to campus to learn about science and technology.

2013 Annual Meeting of the MOST-USDA Joint Research Center for Food Safety

CFSE director Lisa Mauer and key scientist Bruce Applegate joined a team from USDA in China to participate in the 2013 Annual Meeting of the MOST-USDA Joint Research Center for Food Safety hosted at Shanghai Xiao Tong University and to attend the Sino-US Symposium on Food Quality in Nanjing, China. In addition to the scientific conferences, the group toured several agricultural production and processing facilities in China to better understand the similarities and differences in food production and handling between the USA and China.

ANNUAL HIGHLIGHTS

Read more at: http://phys.org/news/2013-10-device-food-pathogen.html

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Annual USDA-CFSE Research Planning MeetingCFSE scientists traveled to USDA-ERRC in Wyndmoor, PA in January 2014 to participate in the annual research planning meeting. In addition to research updates and planning of collaborative studies, the CFSE pathogen detection technologies were demonstrated in the USDA laboratories.

ANNUAL HIGHLIGHTS

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RESEARCH PROJECT OVERVIEW

Dr. Rashid BashirProject Title: BIOCHIP: Microfabrication of biochips able to concentrate, quantify, and detect pathogenic bacteria from food using PCR

Project Description: Biochips are miniature laboratories, the size of a postage stamp, that are able to perform many simultaneous functions to rapidly screen numerous samples. Polymerase chain reaction (PCR) is a commonly used laboratory technique in which a little DNA from a target bacterium is used to generate thousands to millions of copies of the DNA sequence. These copies can then be detected using the biochips. Coupling PCR with the biochip technology results in a rapid detection method for identifying pathogenic bacteria in a food sample.

Project Highlight: This PCR on a biochip system is a fast, portable, and inexpensive method for on-site testing of foodborne pathogen contamination.

Dr. Bruce ApplegateProject Title: PHAGE: Development of bacteriophages for the detection of E. coli O157:H7 and other pathogenic bacteria in food

Project Description: Bacteriophages are viruses that are only able to infect bacteria. The goal of this project is to have the bacteriophages produce and emit light when they have infected a target bacterium. The light can then be detected to rapidly identify if a harmful pathogenic bacteria, such as E. coli O157:H7, was present in a food sample.

Project Highlight: This bacteriophage technology does not require new sample preparation procedures or complicated concentration schemes. By integrating into existing food safety laboratory procedures, it can be implemented into current protocols with ease.

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Dr. Amanda DeeringProject Title: PLANT-PATHOGEN INTERACTIONS

Project Description: Fresh produce has been implicated in numerous outbreaks in recent years. This project aims to better understand the interactions between human pathogenic bacteria and plant tissues, using classical microbiological techniques and developing immunocytochemical methods to assess the ability of pathogens to persist, grow, and localize on and within plants.

Project Highlight: By better understanding how human pathogenic bacteria interact with plants, prevention methods can be developed to aid in providing safe fresh produce to consumers.

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RESEARCH PROJECT OVERVIEW Drs. Arun Bhunia, J. Paul Robinson, Euiwon Bae, and Bartek RajwaProject Title: BARDOT: Development of light scattering technologies for the identification of pathogenic bacteria

Project Description: The BARDOT sensor (Bacterial Rapid Detection using Optical light scattering Technology) is a noninvasive label-free detection and identification system that works by passing a laser beam through each bacterial colony present on a Petri-dish. This generates a light scatter signature that is specific to each bacterium (analogous to a human fingerprint) and enables the identification of bacterial pathogens in food samples.

Project Highlight: BARDOT is a user-friendly high throughput detection device that can rapidly screen food samples for the presence of harmful bacterial pathogens to enhance food safety, reduce foodborne outbreaks, and save lives.

Credit: http://mbio.asm.org/content/5/1/e01019-13

Dr. Joseph IrudayarajProject Title: RAMAN SENSOR: Development of a highly sensitive enhanced Raman spectrosensor for the identification of foodborne pathogens

Project Description: Surface enhanced Raman spectroscopy (SERS) can be used to detect bacteria in foods by identifying different spectra that are unique to each bacterium, much like fingerprint analysis. The SERS signal enhancement enables the detection of low concentrations of pathogenic bacteria in food samples. Sensitive fluorescent assays were also developed to quickly detect low numbers of pathogenic bacteria in foods.

Project Highlight: Two sensitive biosensor techniques, using Raman spectroscopy and fluorescent immunoassays, are portable and useful for rapid onsite detection of foodborne pathogens.

Dr. Michael LadischProject Title: C3D: CONCENTRATION OF PATHOGENIC BACTERIA FROM FOOD SAMPLES

Project Description: An important first step in pathogen detection is the ability to quickly separate the microorganisms from large food samples without harming them. The complexity of food samples makes this a particular challenge. The C3D technique combines mechanical shearing and enzyme treatments with rapid microfiltration through special membranes. After passing through the C3D, the small volume of concentrated sample contains the bacteria that were present in the initial food sample. This is important when the final detection method is only capable of analyzing small volumes, as is the case for the PCR and biochip techniques.

Project Highlight

We could think of C3D as a quick way to find a few needles in a haystack, and then be able to test the needles to find out what kind they are.

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Dr. Robert PruittProject Title: BACTERIAL COMMUNITY ANALYSIS

Project Description: Our project uses molecular methods to validate both classical selective enrichment methods and more modern laser scatter pattern detection methods for their ability to accurately identify specific human pathogenic bacteria. These techniques are being used to characterize the complete bacterial community found on fresh produce that has been associated with foodborne illness outbreaks.

Project Highlight: By using modern DNA sequencing technology to better understand the communities of bacteria that naturally associate with plants and how those communities can sometimes be infiltrated by human pathogens, we can better understand situations that result in outbreaks of foodborne illness.

RESEARCH PROJECT OVERVIEW

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Dr. Haley OliverProject Title: VIRULENCE CAPACITY

Project Description: The long-term goal of this effort is to assess genetic variation and virulence capacity within Salmonella Heidelberg and Listeria monocytogenes. This work will provide insight into characteristics that make certain strains more likely to cause human disease and may help determine which should be considered higher priority for regulation and detection.

Project Highlight: Over the last year, we found that Listeria monocytogenes isolates from retail deli environments have a higher likelihood to potentially cause disease when compared to isolates from retail foods, suggesting that cross-contamination from the deli environment to foods poses a significant public health risk.

Photo by: Brian Powell

SCIENTIFIC TEAM RECOGNITIONS CFSE scientific team recent recognitions

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Director Dr. Lisa Mauer was honored with the 2014 Marcel Loncin Research Prize, one of the top awards from the Institute of Food Technologists. The award honors and provides research funding to a scientists conducting basic chemistry/physics/engineering research applied to food processing and improvement of food quality.

Dr. Michael (Mike) Ladisch was named a Fellow of the National Academy of Inventors, a recognition given to those “who have demonstrated a highly prolific spirit of innovation in creating or facilitating outstanding inventions that have made a tangible impact on quality of life, economic development, and the welfare of society”.

Dr. Haley Oliver was recognized by Vance Publishing in their inaugural 40 Under 40 in Agriculture Awards, which were given to a group of 40 young agriculture leaders for their leadership and commitment in advancing the cause to double food production by 2050. The candidates were nominated because of their passion for agriculture, demonstrated involvement in the food industry, and their notable record of achievement.

Dr. Robert (Bob) Pruitt was named director of Purdue University’s new Center for Molecular Agriculture. The center is part of Purdue’s Plant Sciences Research and Education Pipeline initiative intended to enhance the university’s position as a world leader in plant research to help feed a rapidly growing world population.

Dr. Rashid Bashir became head of the Department of Bioengineering at the University of Illinois at Urbana-Champaign. In this role he plans to take advantage of the “tremendous opportunities to address grand challenges in medicine and biology using engineering approaches to have a profound impact on society and the world at large”.

There were also numerous awards received by CFSE students at local, national, and international scientific meetings. For example, the presentation on the bacteriophage modification to enable low-cost, sensitive detection of E. Coli O157:H7 by Carla Rosenfield was awarded the best poster prize in the area of New Analytical Means of Verifying the Integrity of the Agri Food Supply Chain at the ASsured, SafE and Traceable food (ASSET) meeting held at Queen’s University, Belfast, Ireland.

SCIENTIFIC PUBLICATIONS1) Bhunia, A. K. 2013. Forthcoming new technologies for microbial detection. p. 414-421. In R.G.L.A.S. Garcia

(ed.), Guide to Foodborne Pathogens Wiley-Blackwell, Oxford, UK.2) Chan, V., Asada, H.H., and R. Bashir. 2014. Utilization and control of bioactuators across multiple length

scales. Lab on a Chip. 14:653-657.3) Cho, I. H., and J. Irudayaraj. 2013. In-situ immuno-gold nanoparticle network ELISA biosensors for pathogen

detection. International Journal of Food Microbiology. 164:70-5.4) Cho, I. H., and J. Irudayaraj. 2013. Lateral-flow enzyme immunoconcentration for rapid detection of Listeria

monocytogenes. Analytical and Bioanalytical Chemistry. 405:3313-9.5) Craig, A. P., A. S. Franca, and J. Irudayaraj. 2013. Surface-enhanced raman spectroscopy applied to food safety.

Annual Review of Food Science and Technology, Vol 4. 4:369-380.6) Damhorst, G. L., C. E. Smith, E. M. Salm, M. M. Sobieraj, H. K. Ni, H. Kong, and R. Bashir. 2013. A liposome-

based ion release impedance sensor for biological detection. Biomedical Microdevices. 15:895-905.7) Dorvel, B., B. Reddy, and R. Bashir. 2013. Effect of biointerfacing linker chemistries on the sensitivity of silicon

nanowires for protein detection. Analytical Chemistry. 85:9493-9500.8) Duarte-Gomez, E. E., Graham, D., Budzik, M., Paxson, B., Csonka, L., Morgan, M., Applegate, B., and M.F.

San Martin-Gonzalez. 2013. High hydrostatic pressure effects on bacterial biolumenscence. LWT - Food Science and Technology. 56:484-493.

9) Ebrahimi, A., P. Dak, E. Salm, S. Dash, S. V. Garimella, R. Bashir, and M. A. Alam. 2013. Nanotextured superhydrophobic electrodes enable detection of attomolar-scale DNA concentration within a droplet by non-faradaic impedance spectroscopy. Lab on a Chip. 13:4248-4256.

10) Kim, H., Bai, N., Bhunia, A.K., King, G.B., Hirleman, E.D., and E. Bae. 2013. An integrated morphology analyzer for characterization of optical properties of bacteria micro-colonies. Journal of Biophotonics. 6:929-937.

11) Lathrop, A. A., Bailey, T.W., Kim, K.P., and A.K. Bhunia. 2014. Pathogen-specific antigen target for production of antibodies produced by comparative genomics. Antibody Technology Journal. 4:1-10.

12) Li, X., Ximenes, E., Amalaradjou, M.A.R., Vibbert, H.B., Foster, K., Jones, J., Liu, X., Bhunia, A., and M. Ladisch. 2013. Rapid sample processing for foodborne pathogen detection via crossflow microfiltration. Applied and Environmental Microbiology. 79:7048-7054.

13) Liu, J., and J. Irudayaraj. 2013. Second harmonic generation correlation spectroscopy for single molecule experiments. Optics Express. 21:27063-27073.

14) Pleitner, A.M., Trinetta, V., Morgan, M.T., Linton, R.L., and H.F. Oliver. 2014. Transcriptional and phenotypic responses of Listeria monocytogenes to chlorine dioxide. Applied and Environmental Micriobiology. 80:2951-2963.

15) Popescu, G., Park, K., Mir, M., and R. Bashir. 2014. New technologies for measuring single cell mass. Lab on a Chip. 14:646-652.

16) Singh, A. K., Bettasso, A.M., Bae, E., Rajwa, B., Dundar, M. M., Forster, M.D., Liu, L., Barrett, B., Lovchik, J., Robinson, J.P., Hirleman, E.D., and A. K. Bhunia. 2014. Laser optical sensor, a label-free on-plate Salmonella enterica colony detection tool. mBio. 5.

17) Singh, A. K., Sarkar, P., Janaswamy, S., Yao, Y., and A.K. Bhunia. 2014. Encapsulation and delivery of antimicrobial compounds. p. 218-236. In I.S. Boziaris (ed.), Novel Food Preservation and Microbial Assessment Techniques CRC Press, Boca Raton, FL.

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18) Zhang, Y., Keum, H., Park, K., Bashir, R., and S. Kim. 2014. Micro-masonry of MEMS sensors and actuators. Journal of Microelectromechanical Systems. 23:308-314.

19) Zhou, Z., Kadarn, U.S., and J. Irudayaraj. 2013. One-stop genomic DNA extraction by salicylic acid-coated magnetic nanoparticles. Analytical Biochemistry. 442:249-252.

• 15th International Conference on BioMedical Engineering – Singapore• 2013 Annual Meeting of the MOST-USDA Joint Research Center for Food Safety - Shanghai, China• 2013 Device Research Conference (DRC) – South Bend, IN• 82nd Annual Meeting, Indiana Dietetic Association – West Lafayette, IN• Advances in Microfluidics & Nanofluidics 2013, University of Notre Dame - South Bend, Indiana• American Chemical Society National Meeting – Dallas, Texas• American Institute of Chemical Engineers Annual Meeting- San Francisco, CA• American Society for Microbiology Annual Meeting- Boston, MA• Annual Meeting Center for Innovative Instrumentation Technology, University of Illinois – Urbana, IL• ASME 2014 3rd Global Congress on NanoEngineering for Medicine and Biology - San Francisco, CA• ASsured, SafE and Traceable food (ASSET) Food Integrity and Traceability Conference 2014, Queen’s University- Belfast, Ireland• Central Institute of Fisheries Education - Kolkata, India• Chinese Academy of Inspection and Quarantine Comprehensive Test (CAIQTest) Test Center- Beijing, China• Great Lakes Vegetable Working Group Annual Meeting – West Lafayette, IN• IEEE EMBS Conference of the IEEE Engineering in Medicine and Biology Society- Osaka, Japan• IEHA 63rd Annual Fall Educational Conference – Fort Wayne, IN• Illinois-Tsinghua Nanotechnology Symposium/CNST 12th Annual Nanotechnology Workshop, University of Illinois – Urbana, IL• Indiana Vegetables Growers Association Annual Meeting – West Lafayette, IN• Institute of Food Technologists Annual Meeting and Food Expo – Chicago, IL• International Association of Food Protection Annual Meeting - Des Moines, Iowa• International Fermented Food Expo (IFFE), Chonbuk National University - Jeonju, Korea• Korea Food Research Institute - SeongNam-SI, Korea• Purdue Council for Agricultural Research, Extension and Teaching Annual State Conference – West Lafayette, IN• Purdue Undergraduate Research Symposium – West Lafayette, IN• Purdue University Honors College Discovery Poster Symposium-West Lafayette, IN• Purdue University Health and Disease: Science, Culture and Policy Graduate Student Poster Competition -West Lafayette, IN• Purdue University Summer Undergraduate Research Fellowship Poster Symposium -West Lafayette, IN• Rapid Methods Europe 2014 - Noordwijkerhout, The Netherlands• Transducers 2013 & Eurosensors XXVII: The 17th International Conference on Solid-State Sensors, Actuators and Microsystems –

Barcelona, Spain• UK Food Standards Agency - London, UK

WORK PRESENTED AT THE FOLLOWING:

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CENTER FOR FOODSAFETY ENGINEERING

Philip E. Nelson Hall of Food Science - 745 Agriculture Mall Drive - West Lafayette, IN 47907

For more information, contact:Dr. Lisa Mauer Professor and Director mauer@purdue.edu 765-494-9111

Dr. Amanda Deering Operations Manager adeering@purdue.edu 765-494-0512

Or visit our website:

www.cfse.purdue.edu

2013 Annual Meeting of the MOST-USDA Joint Research Center for Food Safety

Dr. Bruce Applegate 765-496-7920 applegate@purdue.edu

Dr. Euiwon Bae 765-494-4762 baee@ecn.purdue.edu

Dr. Rashid Bashir 217-333-3097 rbashir@illinois.edu

Dr. Arun Bhunia 765-494-5443 bhunia@purdue.edu

Dr. Amanda Deering 765-494-0512 adeering@purdue.edu

Dr. Joseph Irudayaraj 765-494-0388 josephi@purdue.edu

Dr. Michael Ladisch 765-494-7022 ladisch@purdue.edu

Dr. Haley Oliver 765-496-3913 hfoliver@purdue.edu

Dr. Robert Pruitt 765-496-6794 rep@purdue.edu

Dr. Bartek Rajwa 765-496-1153 brajwa@purdue.edu

Dr. J. Paul Robinson 765-494-6449 wombat@purdue.edu

Dr. Manpreet Singh 765-494-0823 manpreet@purdue.edu

Center for Food Safety Engineering Key Scientists