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Center for foodsafety engineering
2012 Newsletter
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.
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The 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 has been 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:
1. C3D: Effective food sampling protocols and filtration techniques to maximize microbial cell separation and concentration in an automated instrument.
2. 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.
3. BARDOT: An image database and refinement of the Bacterial Rapid Detection using Optical scattering Technology (BARDOT) system for microcolony detection and identification, shortening the detection time for L. monocytogenes, select Salmonella serovars, and STEC.
4. 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.
5. Raman Sensor: A Raman biosensing platform for detecting pathogens with a single cell sensitivity.
There were many highlights and banner accomplishments during 2011-2012. The CFSE participated in the Sixth International Forum on Food Safety and 2011 Annual Meeting of the MOST-USDA Joint Research Center for Food Safety in Shanghai, China, with additional visits and presentations to Jiangnan University in Wuxi, China, and Huazhong University in Wuhan, China. The BARDOT technology, licensed and produced by Advanced Bioimaging Systems, was featured in national and international trade shows. A record number of invention disclosures and patent applications were also completed. 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 food supply.
Note from the director
DirectorDr. Lisa MauerProfessor,Department of Food Science
tABLe of coNteNtSoutreaCh 3
researCh ProjeCt highlights 4
sCientifiC PubliCations and Presentations 6
Work Presented 7
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outreAch
PAthoGeN trAceABiLity uSiNG BArdot
In an effort to help people working in the food industry, from plant production personnel to sales and marketing personnel, understand the need for and application of technologies being developed through the Center for Food Safety Engineering (CFSE), the Center and USDA offered a live 90-minute webcast hosted by the Institute of Food Technologists (IFT) titled, “Rapid Detection Technologies for Foodborne Pathogens.”
Foodborne illness in the United States is an important health concern. Outbreaks highlight key deficiencies in the food distribution chain, and one of them is the lack of traceability methodology to track contaminated food back to the source in an effective manner. One of the four critical areas under the FDA Food Safety Modernization Act is the need for better detection of and response to food safety problems. Common food testing strategies include traditional bacterial culture/plating of food samples to test for bacterial pathogen contamination, as well as advanced testing options including DNA and immunology-based assays. While these assays are sensitive and reliable, they suffer from key limitations including: the inability to reliably distinguish viable
bacteria in food samples, the time required (3-7 days) to obtain a result, etc.
CFSE and USDA scientists have been working to develop technologies that will enhance the ability to sample and test increasing amounts of foods as well as improve the detection and analysis of foodborne pathogens.
The webcast featured CFSE scientists Drs. Lisa Mauer, Arun Bhunia, Michael Ladisch, and Rashid Bashir, and USDA-ARS scientist Dr. George Paoli. The scientists highlighted several technologies including a cell concentration and recovery device; light scanner imaging technology called BARDOT, for detection and analysis of foodborne pathogens; high-throughput biosensors for multiplexed foodborne pathogen detection; and USDA research activities.
The schematic below developed by Dr. J. Paul Robinson illustrates how much faster foodborne pathogens can be detected by using BARDOT technologies, developed by the CFSE, compared to traditional methods.
With BARDOTBArdot reSeArch ceNterS
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dr. rashid bashirProject titleBiochip: Microfabrication of biochips that are able to concentrate, quantify, and detect pathogenic bacteria from food using PCR
Project DescriPtioNBiochips are miniature laboratories, the size of a postage stamp, that are able to perform many simultaneous functions to rapidly screen many samples. Polymerase chain reaction (PCR) is a common laboratory technique used where a single piece or a few copies of a piece of DNA are used to generate thousands to millions of copies of a particular DNA sequence from a target bacterium. Coupling PCR with biochip technology results in a rapid detection method for identifying pathogenic bacteria in a food sample.
Project HigHligHtThis PCR on a biochip system is a fast, portable, inexpensive method for on-site testing of foodborne pathogen contamination.
reSeArch Project hiGhLiGhtSdr. MiChael ladisChProject titleC3D: Concentration of pathogenic bacteria from food samples
Project DescriPtioNSample preparation methods that quickly concentrate the microorganisms present in a large food sample with minimal impact on their viability are an important first step in pathogen detection. The complexity of food samples makes this a particular challenge. In this technique, the combined use of mechanical shearing and enzyme treatments enables rapid microfiltration of the bacteria through special membranes. The small volume of concentrated sample produced represents all of the bacteria present in the large food sample. This is important when the final detection method is only capable of analyzing small volumes, such as for the polymerase chain reaction (PCR) technique.
Project HigHligHtThe concentration of pathogenic bacteria from food samples will allow for the rapid detection of the bacterial pathogens near the source so that interventions may be implemented sooner to reduce the distribution of unsafe food to consumers.
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. bruCe aPPlegateProject titlePhage: Development of bacteriophages for the detection of E. coli O157:H7 and other pathogenic bacteria in food
Project DescriPtioNBacteriophages are viruses that are only able to infect bacteria. The goal of this project is to have the bacteriophage 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 HigHligHtThis technology does not require new sample preparation procedures or complicated concentration schemes. By integrating into existing food safety laboratory procedures, and with an estimated cost of 11 cents per assay, as compared to several dollars per test for current methods, it is a cost effective means of detecting pathogenic bacteria in a food sample.
dr. arun bhuniaProject titleBARDOT: Development of light scattering technologies for the identification of pathogenic bacteria
Project DescriPtioNA laser sensor is used to instantly screen bacterial
colonies on a Petri-dish for early pathogen detection. The sensor, designated BARDOT (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 allows for the identification of bacterial pathogens in food samples.
Project HigHligHtBARDOT 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.
dr. josePh irudayarajProject titleRaman Sensor: Development of a highly sensitive enhanced Raman spectrosensor for the identification of foodborne pathogens
Project DescriPtioNSurface enhanced Raman spectroscopy (SERS) can be used to detect bacteria in a food system 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.
Project HigHligHtRaman spectroscopy-based biosensors can detect foodborne pathogens and the portable system being developed will enable on-site detection.
Phage infecting the bacterium The light emitting response can be quantified for detection
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1. Bae, E., Patsekin, V., Rajwa, B., Bhunia, A.K., Holdman, C., Davisson, V.J., Hirleman, E.D., Robinson, J.P. 2012. Development of a microbial high-throughput screening instrument based on elastic light scatter patterns. Review of Scientific Instruments, 83: 044304.
2. Bhunia, A.K., Bae, E., Rajwa, B., Robinson, J.P., Hirleman, E.D. 2011. Utilization of optical forward scatter image biological database: food-borne pathogen colony differentiation and detection. In: Omics Technologies and Microbial Modeling in Food-borne Pathogens Study. Editors: Yan, X., Juneja, V., Huang, L. DEStech Publications, Inc. Lancaster, PA. p. 599-624.
3. Davis, R., Deering, A., Burgula, Y., Mauer, L.J., Reuhs, B.L. 2012. Differentiation of live, dead, and treated cells of E. coli O157:H7 using FT-IR spectroscopy. Journal of Applied Microbiology, 112(4):743-751.
4. Davis, R. and L.J. Mauer. 2011. Subtyping of Listeria monocytogenes at the haplotype level by Fourier transform infrared (FT-IR) spectroscopy and multivariate statistical analysis. International Journal of Food Microbiology, 150(2-3):140-149.
5. Davis, R., Paoli, G., Mauer, L.J. 2012. Evaluation of Fourier transform infrared (FT-IR) spectroscopy and chemometrics as a rapid approach for subtyping E. coli O157:H7 isolates. Food Microbiology, 31:181-190.
6. Deering, A.J., Mauer, L.J., Pruitt, R.E. 2012. Internalization of E. coli O157:H7 and Salmonella spp. in plants: A review. Food Research International, 45:567-575.
7. Deering, A.J., Pruitt, R.E., Mauer, L.J., Reuhs, B.L. 2011. Identification of the cellular location of internalized Escherichia coli O157:H7 in mung bean, Vigna radiata, by immunocytochemical techniques. Journal of Food Protection, 4(8):1224-1230.
8. Deering, A.J., Pruitt, R.E., Mauer, L.J., Reuhs, B.L. 2012. Examination of the internalization of Salmonella serovar Typhimurium in peanut,
Arachis hypogaea, using immunocytochemical techniques. Food Research International, 45:1037-1043.
9. Gehring, A.G., Albin, D.M., Bhunia, A.K., Kim, H., Reed, S.A., Tu, S.I. 2012. Mixed culture enrichment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica, and Yersinia enterocolitica. Food Control, 26: 269-273.
10. Goodridge, L.D., Fratamico, P., Christensen, L.S., Hoorfar, J., Griffiths, M., Carter, M., Bhunia, A.K., O’Kennedy, R. 2011. Strengths and shortcomings of advanced detection technologies. In: Rapid Detection, Characterization and Enumeration of Foodborne Pathogens. Editor, J. Hoorfar. ASM Press, Washington DC. p. 15-45.
11. Hoorfar, J., Christensen, B.B., Pagotto, F., Rudi, K., Bhunia, A.K., Griffith, M. 2011. Future trends in rapid methods. In: Rapid Detection, Characterization and Enumeration of Foodborne Pathogens. Editor, J. Hoorfar. ASM Press, Washington DC. p. 413-420.
12. Huff, K., Aroonnual, A., Fleishman-Littlejohn, A.E., Rajwa, B., Bae, E., Banada, P.P., Patsekin, V., Hirleman, E.D., Robinson, J.P., Richards, G.P., Bhunia, A.K. 2012. Light-scattering sensor for real-time identification of Vibrio parahaemolyticus, V. vulnificus, and V. cholerae colonies on solid agar plate. Microbial Biotechnology, Published online 21 May, DOI: 10.1111/j.1751-7915.2012.00349.x
13. Jagadeesan, B., Fleishman-Littlejohn, A.E., Amalaradjou, M.A.R., Singh, A.K., Mishra, K., La, D., Kihara, D., Bhunia, A.K. 2011. N-terminal Gly224–Gly411 domain in Listeria adhesion protein interacts with host receptor Hsp60. PLoS One, 6: e20694.
14. Jaradat, Z.W., Rashdan, A.M., Ababneh, Q.O., Jaradat, S.A., Bhunia, A.K. 2011. Characterization of surface proteins of Cronobacter muytjensii using monoclonal antibodies and MALDI-TOF Mass spectrometry. BMC Microbiology, 11:148.
ScieNtific PuBLicAtioNS ANd PreSeNtAtioNS
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15. Koo, O.K., Amalaradjou, M.A.R., Bhunia, A.K. 2012. Recombinant probiotic expressing Listeria adhesion protein attenuates Listeria monocytogenes virulence in vitro. PLoS One, 7: e29277.
16. Malorny, B., Bhunia, A.K., Aarts, H.J.M., Löfström, C., Hoorfar, J. 2011. Salmonella in pork, beef, poultry and egg. In: Rapid detection, characterization and enumeration of food-borne pathogens. Editor, J. Hoorfar, ASM Press, Washington DC. p. 179-194.
17. Mura, S., Greppi1, G., Marongiu, M.L., Roggero, P.R., Ravindranath, S.P., Mauer, L.J., Schibeci, N., Perria, F., Piccinini, M., Innocenzi, P., Irudayaraj,
J. 2012. FTIR nanobiosensors for Escherichia coli detection. Beilstein Journal of Nanotechnology, 3:485-492.
18. Robinson, J.P., Rajwa, B.P., Bae, E., Patsekin, V., Roumani, A.M., Bhunia, A.K., Dietz, J.E., Davidson, V.J., Dunder, M.M., Thomas, J., Hirleman, E.D. 2011. Using scattering to identify bacterial pathogens. Optics and Photonics News, v. 22: 21-27.
19. Salm, E., Liu, Y.S., Machwiany, D., Morisette, D.T., He, Y., Razouk, L., Bhunia, A.K., Bashir, R. 2011. Electrical detection of dsDNA and polymerase chain reaction amplification. Biomedical Microdevices, 13: 973-982.
iNterNAtioNAL:• AnnualMeetingofMOST-USDAJointResearch
Center for Food Safety - Shanghai, China• BiosensorTechnologiesforPathogenandToxin
Detection, Jiangnan University - Wuxi, China• InternationalSymposiumonRecentAdvancesin
Food Analysis - Prague, Czech Republic• TechnicalUniversityofDenmark-Copenhagen,
Denmark• KoreaAdvancedInstituteofScienceand
Technology - Daejon, Korea• KoreaInstituteofScienceandTechnology-Seoul,
Korea• AntimicrobialResearch-Beijing,China• FoodSafetyForum-Shanghai,China• NoguchiMemorialInstituteForMedicalResearch
College of Health Sciences – Legon, Ghana
domeStic:• AmericanChemicalSocietyMeeting-SanDiego,CA• AIMBE20thAnnualEvent-Washington,DC• NovozymesNorthAmerica,Inc.-Franklinton,NC• CPIASRInternationalForum,PurdueUniversity-
West Lafayette, IN• SocietyofAcuteWoundCare–LasVegas,NV• InstituteofFoodTechnologists(IFT)Annual
meeting – Las Vegas, NV• AmericanSocietyforMicrobiologyGeneral
Meeting - San Francisco, CA• TheSocietyofSigmaXi,PurdueUniversity
Chapter - West Lafayette, IN• GeorgiaInstituteofTechnology-Atlanta,GA• AmericanSocietyofAgriculturalandBiological
Engineers - Louisville, KY• IowaStateUniversity–Ames,IA• InternationalConferenceforFoodSafetyand
Quality – Chicago, IL• IEEEPhotonicsConference-SanFrancisco,CA• Nano-BioCollaborativeInternationalConference
2012 - Tampa, FL• UniversityofKentucky-Lexington,KY• TexasA&MUniversity-CollegeStation,TX
Work Presented at the FolloWing:
Center for Food Safety EngineeringPhilip E. Nelson Hall of Food Science745 Agriculture Mall DriveWest Lafayette, IN 47907
For more information, contact:Dr. Lisa MauerProfessor and [email protected]
Dr. Amanda DeeringOperations [email protected]
or visit our website:www.cfse.purdue.edu
Sixth iNterNAtioNAL forum oN food SAfety ANd 2011 ANNuAL meetiNG of moSt-uSdA joiNt reSeArch ceNter for food SAfety. ShANGhAi, chiNA.SePt 27-29, 2011
dr. bruCe aPPlegate765-496-7920
dr. euiWon bae765-494-4762
dr. rashid bashir217-333-3097
dr. arun bhunia765-494-5443
dr. josePh irudayaraj765-494-0388
cfSe Key ScieNtiStS
dr. MiChael ladisCh765-494-7022
dr. bartek rajWa765-494-0757
dr. j. Paul robinson765-494-0757
dr. eduardo XiMenes765-494-6695
