NOVEL APPROACHES FOR THE DETECTION AND CONTROL OF FOODBORNE BACTERIAL AND VIRAL PATHOGENS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0221815
• Project No.: TEN00391
• Project Start Date: Oct 1, 2009
• Project End Date: Sep 30, 2014

Project Director
D'Souza, D.

Recipient Organization
UNIVERSITY OF TENNESSEE
2621 MORGAN CIR
KNOXVILLE,TN 37996-4540

Performing Department
Food Science & Technology

Non Technical Summary
Foodborne bacterial and viral diseases are a growing concern to the agricultural and food industry. These disease outbreaks result in significant economic losses and remain a public health concern. The proposed project aims at rapidly detecting these pathogens in the food environment using novel molecular tools. In addition, the project aims at developing mitigation and control strategies to prevent transmission of foodborne disease outbreaks and to protect publich health.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
712	4010	1040	10%
712	4010	1100	10%
712	4030	1040	10%
712	4030	1101	10%
712	4099	1040	10%
712	4099	1100	10%
722	4099	1040	10%
722	4010	1100	10%
722	4030	1101	10%
722	4030	1040	10%

Knowledge Area
722 - Zoonotic Diseases and Parasites Affecting Humans; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
4099 - Microorganisms, general/other; 4010 - Bacteria; 4030 - Viruses;

Field Of Science
1101 - Virology; 1100 - Bacteriology; 1040 - Molecular biology;

Keywords

foodborne viruses

salmonella

e. coli

detection

control

transmission

Goals / Objectives
(1) Develop rapid and robust detection technologies for identification and tracking of foodborne pathogens (bacterial and viral) in order to prevent their transmission, growth and survival in the environment; (2) Develop novel intervention strategies to control foodborne bacterial and viral pathogens in the environment and evaluate their efficacy on food contact surfaces or directly in foods; (3) Determine methods to control and prevent outbreaks by using fundamental approaches to understand gene expression profiles of foodborne bacteria under various conditions of growth and stress encountered in the food processing environments; (4) Utilize functional genomics and proteomic technologies for improved detection and development of novel strategies for controlling foodborne pathogens

Project Methods
This project aims at developing robust diagnostic tools for the rapid and sensitive detection of foodborne pathogens to reduce the risk of disease spread via consumption of food. Improved molecular and other novel approaches will be researched for application in food systems. Novel processing and control technologies such as high pressure homogenization, high-intensity ultrasound, washes or rinses, including addition of bioactive and ecologically-friendly methods will be used to inactivate or reduce the pathogen load in food matrices or the food environment. Understanding gene and protein expression for the control and prevention of outbreaks will help in the basic fundamental understanding of bacterial pathogenesis as well.

Progress 10/01/09 to 09/30/14

Outputs
Target Audience: University students and attendees nationwide at regional and national conferences/meetings and symposia. Industry attendees at grantees meetings and annual reporting meetings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training of undergraduate students, graduate students, and staff along with professional meeting attendancewere provided. How have the results been disseminated to communities of interest? At professional meetings, annual grantee's meeting, lecturesand workshops. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The investigator's research during this reporting period focused on (1) Improvedmolecularassays to detect human norovirus from clinicalsamples(2)Novel processing and intervention strategies to control foodbornebacterial and viral pathogensincluding natural antimicrobials. Natural bioactive components of plants were also investigated forantiviral and antibacterial activity. (3) Fundamental omic-basedand next-generation sequencing approachesto determineappropriate control and mitigation strategies in food environments (in collaboration with researchers atUT FST and UT-Microbiology Departments). Robust detection of foodborne pathogens from food and the food environment help to curb foodborne disease outbreaks.Rapid detection helps protect public health due to decrease in foodborne disease incidence, decreased recalls and decreaseeconomical losses to agriculture. Novel natural ecofriendly intervention strategies that help to sustain the environment andsimultaneously enhance safety of food have potential applications in the food industry. Natural plantbioactive components that have nutritional benefits with added antimicrobial properties can be used as/in functional foods andbeverages or as natural therapeutic agents. Cost-effective, ecofriendly, non-toxic, "green" approaches can be used asalternates to enhance produce safety and also protect the environment. Fundamental "omics" and next-generationsequencing helps understand the basis of foodborne bacterial survival and control in the food environment to improve theoverall safety of foods and the food environment as well as public health.

Publications

• Type: Journal Articles Status: Published Year Published: 2014 Citation: Bozkurt H, D'Souza DH, Davidson PM. A comparison of the thermal inactivation kinetics of human norovirus surrogates and hepatitis A virus in buffered cell culture medium. Food Microbiol. 2014 Sep;42:212-7.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Bozkurt, H., D. H. DSouza, and P. M. Davidson. 2014. Thermal inactivation kinetic modeling of human norovirus surrogates in blue mussel (Mytilus edilus) homogenate. Int J Food Microbiol. Feb 17;172:130-6.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: DSouza DH. 2014. Phytocompounds for the control of human enteric viruses. Current Opinion in Virology Feb;4:44-9.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Joshi, S., A. Howell, and D. H. DSouza. 2014. Cronobacter sakazakii reduction by blueberry proanthocyanidins. Food Microbiology 39: 127-131.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Bozkurt, H., D. H. D'Souza, and P. M. Davidson. 2014. Thermal inactivation of Hepatitis A virus in spinach. International Association of Food Protection, 103rd Annual Meeting, Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Chauhan, S*, S. Joshi*, and D. H. D'Souza. 2014. Salmonella enterica reduction by grape seed extract in apple juice. UT EUReCA Symposium.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: D'Souza, D. H., L. Dice, and P. M. Davidson. 2014. Aqueous extracts of Hibiscus sabdariffa to control Aichi virus. Internationl Society for Food and Environmental Virology Mtg, Corfu, Sept.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Joshi S. S.*, A. B. Howell, and D. H. D'Souza. 2014. Blueberry juice and blueberrry polyphenols against human norovirus surrogates. ASM Annual Meeting, Boston, MA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Joshi, S. S.*, L. Dice, and D. H. D'Souza. 2014. Hibiscus sabdariffa Aqueous Extract Against Human Norovirus Surrogates. American Society for Microbiology Annual Mtg, Boston, MA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Joshi, S.*, A. B. Howell, and D. H. D'Souza. 2014. Blueberry polyphenols for hepatitis A virus control. IAFP.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: M. Valladares, D. H. D'Souza, and P. M. Davidson. 2014. Comparison of the thermal inactivation parameters Shiga-toxin producing E. coli O157:H7 and Non-O157 E. coli in turkey deli meat and ground beef. International Association of Food Protection, 103rd Annual Meeting, Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Sewlikar, S*, and D. H. D'Souza. 2014. Quillaja extracts against emerging O157 STECs and non-O157 STECs. International Association for Food Protection, Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Sewlikar, S. S.*, A. B. Howell, and D. H. D'Souza. 2014. Dose and time-dependence of cranberry proanthocyanidins against human norovirus surrogates. IFT Annual Mtg, New Orleans, LA.

Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training of graduate students, undergraduate students and technical staff. How have the results been disseminated to communities of interest? At professional meetings, annual grantee's meeting and workshops. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The investigator’s research during this reporting period focused on objectives 1, 2, and 3. (1) Improved RT-LAMP isothermal assays and real-time RT-PCR assays were optimized to detect human norovirus from clinical isolates. Human norovirus surrogate studies in juices were also carried out. (2) Improved processing and intervention strategies to control foodborne bacterial and viral pathogens including natural antimicrobials were evaluated. In collaboration with researchers at Rutgers State University of New Jersey, natural alternatives such as novel bioactive components of plants were also investigated for antiviral and antibacterial activity. (3) Fundamental omic-based approaches using next-generation sequencing to determine appropriate control and mitigation strategies in food environments were also investigated in collaboration with researchers at UT FST and UT-Microbiology Departments. Rapid and robust detection of foodborne pathogens from food and the food environment help to prevent foodborne outbreaks. Rapid detection helps protect public health due to decrease in foodborne disease incidence, decreased recalls and decrease economical losses to agriculture. Novel natural ecofriendly intervention strategies that help to sustain the environment and simultaneously enhance safety of food have potential applications in the food industry and in organic farming. Natural plant bioactive components that have nutritional benefits with added antimicrobial properties can be used as/in functional foods and beverages or as natural therapeutic agents. Cost-effective, ecofriendly, non-toxic, “green” approaches can be used as alternates to enhance produce safety and also protect the environment. Fundamental “omics” and next-generation sequencing helps understand the basis of foodborne bacterial survival and control in the food environment to improve the overall safety of foods and the food environment as well as public health.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: Su, X., and D. H. DSouza. 2013. Grapeseed extract for foodborne virus reduction on produce. Food Microbiology 34(1):1-6.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Su X, D'Souza DH. 2013. Naturally occurring flavonoids against human norovirus surrogates. Food Environ Virol. 2013 Jun;5(2):97-102.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Howell, A. B, D. H. DSouza. 2013. The Pomegranate: Effects of bacteria and viruses that affect human health. Evidence-based complementary and alternative medicine 2013:606212. doi: 10.1155/2013/606212. Epub 2013 May 20.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Diaz, S., S. Pendleton, I. Hanning, and D. H. DSouza. 2013. Next generation sequencing to address food safety and poultry issues. Poultry Sci (invited and accepted). Poultry Sci. 2013 Feb; 92(2):562-72.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Gehring, AG, C. Barnett, T. Chu, C. Debroy, D. DSouza, S. Eaker et al. 2013. A high-throughput antibody-based microarray typing platform. Sensors. (open access on-line).
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Bozkurt, H., D. H. DSouza, and P. M. Davidson. 2013. Determination of the thermal inactivation kinetics of human norovirus surrogates, feline calicivirus and murine norovirus. J Food Prot. 76(1):79-84.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sanchez S, S. Joshi, A. Andino, F. Gil, D. DSouza, I Hanning. Grape seed extract as a feed additive reduces Salmonella colonization in broiler chicks. Conference of Research Workers in Animal Diseases, Chicago, Nov 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Joshi, S., A. Howell, and D. H. DSouza. Blueberry juice against human enteric virus surrogates. ASM KY-TN Branch meeting, Oral Presentation, 3rd place Award, October, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sewlikar S. and D. H. DSouza. Quillaja extracts against E. coli O157:H7 and emerging non-O157 STECS. Oral Presentation, ASM KY-TN Branch Meeting, Bowling Green, KY, October 2013
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sanchez, S. C. Yard, F. Gil, D. DSouza, I. Hanning. Preharvest sampling. Oral Presentation, ASM KY-TN Branch Meeting, Bowling Green, KY, October 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: D. DSouza, S. Joshi, L. Dice. Quillaja saponoria extracts against human norovirus surrogates. Fifth Calicivirus conference, Beijing, China, October, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: B. Ownley, N. Labbe, K. Gwinn, N. M. Dee, P. B. Filson , A. L. Vu, D. H. DSouza, N. Moustaid-Moussa. Development of switchgrass extractives as a value-added biopesticide against bacterial spot (Xanthomonas perforans) of tomato. American Plant Pathologists-MSA, Austin, TX, Aug, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: DSouza, DH, X. Su, D. A. Golden. Non-thermal cold plasma for the inactivation of human norovirus surrogates on stainless surfaces. IAFP, Charlotte, NC. Oral presentation, July 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Joshi, S., A. B. Howell, D. H. DSouza. Reduction of Cronobacter sakazakii by blueberry juice and blueberry proanthocyanidins. IAFP, Charlotte, NC, Poster, July, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cicco, A., A. B. Howell, D. H. DSouza. Comparison of grapeseed extract, pomegranate polyphenols, and cranberry extracts against Salmonella enterica serovars Typhimurium and Enteritidis. IAFP, Charlotte, NC, Poster Undergraduate Competition, 2nd Place Award, July 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Leister S, H. Bozkurt, D. H. DSouza, P. M. Davidson. 2013. Thermal inactivation of human norovirus surrogates in blue mussels IAFP, poster presentation. July 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Joshi, S., A.B. Howell, D. H. DSouza. Time-dependent effects of GSE against human norovirus surrogates. ASM Poster, Denver, Colorado, May 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cicco, A., A. B. Howell, D. H. DSouza. Plant extracts against Cronobacter sakazakii. ASM Poster, Denver, Colorado, May 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cong, C., D. H. DSouza. Sodium metasilicate for the inactivation of human norovirus surrogates. IFT, Chicago, July, 2013 Poster
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cong, C. D. H. DSouza. Effects of benzalkonium chloride against human norovirus surrogates using carrier tests. IFT, Chicago, July, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: D. DSouza. Human norovirus surrogates in food safety research. IFT, July 2013. Oral presentation. Symposium (invited speaker).

Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: The investigator's research during this reporting period focused on objectives 1, 2, and 3. (1) Rapid molecular-based technologies for Salmonella detection were applied to food environments to enable tracking of outbreaks. Improved RT-LAMP isothermal assays and real-time RT-PCR assays were optimized to detect human norovirus clinical isolates. Human norovirus studies in juices and juice blends were also carried out. (2) Improved processing and intervention strategies to control foodborne bacterial and viral pathogens including natural antimicrobials were evaluated. In collaboration with researchers at Rutgers State University of New Jersey, and University of Rochester, natural alternatives such as novel bioactive components of plants were also investigated for antiviral and antibacterial activity. High pressure homogenization and ultrasound to inactivate Salmonella and human norovirus surrogates were researched in collaboration with researchers within FST-UT. (3) Fundamental transcriptomic-based approaches to determine appropriate control and mitigation strategies in food environments were also investigated in collaboration with researchers at FST-UT and UT-Microbiology. PARTICIPANTS: Katie Horm, graduate student Chayapa Techathuvanan, graduate student Xiaowei Su, Research Associate Cong Cao, graduate student Hayriye Bozkurt, graduate student Radha Ganapathy, graduate student Barbara Gillespie, Research Associate TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Robust, sensitive, specific and timely detection of foodborne bacterial and viral pathogens from food commodities and the food environment aid in the prevention of foodborne outbreaks. Rapid detection helps protect public health due to decreased foodborne disease outbreaks with decreased recalls and decrease economical losses to agriculture. Novel natural ecofriendly intervention strategies that help to sustain the environment and simultaneously enhance safety of food products have potential applications in the food industry and in organic farming. Natural plant bioactive components that have nutritional benefits with added antimicrobial properties can be used as/in functional foods and beverages or as natural therapeutic agents. Cost-effective, ecofriendly, non-toxic, "green" approaches can be used as alternates to enhance produce safety and also protect the environment. Fundamental "omics" helps understand the basis of foodborne bacterial survival and control in the food environment to improve the overall safety of foods and the food environment as well as public health.

Publications

• Horm, K, PM Davidson, F Harte, D. H. DSouza. 2012. Survival and Inactivation of Human Norovirus Surrogates in Blueberry Juice by High Pressure Homogenization. Foodborne Pathog Dis. Nov;9(11):974-9.
• Horm, KM, F Harte, D. H. DSouza. 2012. Human Norovirus Surrogate Reduction in Milk and Juice Blends by High Pressure Homogenization. J Food Prot. Nov;75(11):1984-90.
• Su, X, D. H. DSouza. 2012. Inactivation of human norovirus surrogates by benzalkonium chloride, potassium peroxymonosulfate, tannic acid, and gallic acid. Foodborne Pathog Dis. 9(9):829-34.
• Davis, R, S Zivanovic. D. H. DSouza, P. M. Davidson. 2012. Effectiveness of chitosan on the inactivation of enteric viral surrogates. Food Microbiol. 2012 Oct;32(1):57-62.
• Su Xiaowei, D. H. DSouza. 2012. Reduction of Salmonella Typhimurium and Listeria monocytogenes on produce by trisodium phosphate LWT-Food Science and Technology, 45 (2): 221-225.
• Su X, Howell AB, D. H. DSouza. 2012. Antibacterial effects of plant-derived extracts on Methicillin-Resistant Staphylococcus aureus. Foodborne Pathog Dis. Jun;9(6):573-8.
• Techathuvanan C, D. H. DSouza. 2012. Reverse-transcriptase loop-mediated isothermal amplification as a rapid screening/monitoring tool for Salmonella enterica detection in liquid whole eggs. J Food Sci. 2012 Apr;77(4):M200-5.
• Su, X, and D. H. DSouza. Time dependent effects of grapeseed extract against hepatitis A virus and human norovirus surrogates. Poster Presentation, 3rd Food and Environmental Virology (FEV) Congress, Lisbon Portugal, Oct., 2012.
• Bozkurt, H., D. H. DSouza, P. M. Davidson. Thermal inactivation of human norovirus surrogates. Poster presentation at the International Association for Food Protection Annual meeting (IAFP), Providence, RI, 2012.
• Chen, S., D. Golden, D. DSouza, F. Critzer. Gene expression analysis of Salmonella Tennessee in a low water activity food. Oral Presentation. IAFP, Providence, RI, 2012.
• Gillespie, B., D. DSouza, et al. Comparison of enrichment media and ELISA for the rapid and sensitive detection of E. coli O157:H7. Poster presentation, IAFP, Providence, RI, 2012
• Cao, C., D. H. DSouza. RT-LAMP for the detection of human norovirus GII. Oral presentation, IAFP, Providence, RI, 2012.
• Cao, C., D. H. DSouza. Chemical inactivation of human norovirus surrogates. Oral presentation, IAFP, Providence, RI, 2012.
• Bozkurt, H., D. H. DSouza, P. M. Davidson. Thermal inactivation kinetics of human norovirus surrogates at one temperature in buffer solutions. Poster Presentation, Institute of Food Technologists Annual Meeting (IFT), Las Vegas, NV, 2012.
• Techathuvanan, C., A. Howell, D. H. DSouza. Blueberry juice against human enteric viruses. Poster Presentation, IFT, Las Vegas, NV, 2012.
• Techathuvanan, C., D. H. DSouza. HIU for the inactivation of Salmonella Enteritidis in LWE. IFT, Las Vegas, NV, 2012.
• Su, X., D. H. DSouza. Chemical inactivation of human norovirus surrogates using BAC and KPMS. Poster Presentation, IFT, 2012
• Ganapathy, R., S. Zivanovic, D. DSouza. Chitosan for the inactivation of human norovirus surrogates. Poster Presentation, IFT, Las Vegas, NV, 2012.
• Su, X., A. Howell, D. H. DSouza. Antibacterial Effects of Plant-Derived Extracts on Methicillin-Resistant Staphylococcus aureus. Poster Presentation, American Society for Microbiology Annual Meeting (ASM), San Francisco, CA, 2012.
• Su, X., D. H. DSouza. Time dependence effects of cranberry proanthocyanidins and pomegranate polyphenols on Hepatitis A virus infectivity. Oral presentation, IAFP, Providence, RI, 2012
• Su, X., D. H. DSouza. Effects of myricetin and L-epicatechin on human norovirus surrogates. Oral presentation, IAFP, Providence, RI, 2012.
• Horm, K., F. Harte, D. H. DSouza. High pressure processing for the inactivation of human norovirus surrogates in milk and juice blends. Oral presentation, IAFP, Providence, RI, 2012

Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: The investigator's research during this reporting period concentrated on objectives 1, 2, and 3. (1) Novel rapid and robust molecular-based technologies for Salmonella detection were applied to eggs to enable tracking and transmission of outbreaks in the food environment. Improved real-time RT-PCR assays to detect human norovirus clinical isolates were optimized and applied to detect human norovirus from produce. Studies to understand the survival of human norovirus surrogates in milk and juices and in order to provide data to assess risk and methods of control were undertaken. (2) Improved intervention strategies to control foodborne bacterial and viral pathogens including natural antimicrobials were evaluated. Novel chemicals were used to determine inactivation of foodborne bacterial and viral pathogens with improved efficacy. In collaboration with researchers at Rutgers State University of New Jersey, and University of Rochester, natural alternatives such as novel bioactive components of plants were also investigated for antiviral activity. Novel processing technologies to inactivate foodborne viral pathogens such as high pressure homogenization and ultrasound to inactivate Salmonella were researched in collaboration with researchers within FST-UT. (3) Fundamental transcriptomic-based approaches to determine appropriate control and mitigation strategies in food environments were also investigated in collaboration with researchers at FST-UT. PARTICIPANTS: Nathan Duray Miller, graduate student Katie Horm, graduate student Chayapa Techathuvanan, graduate student Xiaowei Su, Research Associate Dragoslava Radin, Fulbright Scholar and Visiting Associate Professor TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Robust, rapid, and sensitive detection methods for foodborne bacterial and viral pathogens from food commodities and the food environment can aid in the prevention of foodborne outbreaks. Rapid detection will help protect public health due to decreased foodborne disease outbreaks with decreased recalls and economical losses to agriculture. Novel natural ecofriendly intervention strategies that help to sustain the environment and simultaneously enhance safety of food products have potential applications in the food industry. Natural plant bioactive components that have nutritional benefits with added antimicrobial properties can be used as/in functional foods and beverages or as natural therapeutic agents. Ecofriendly, non-toxic, "green" approaches can be used as alternates to enhance produce safety and also protect the environment. Using fundamental "omics" approaches, the basis of foodborne bacterial survival and control in the food environment can be delineated to improve the overall safety of foods and the food environment.

Publications

• Techathuvanan C, DSouza DH. 2011. Optimization of rapid Salmonella enterica detection in liquid whole eggs by SYBR green I-based real-time reverse transcriptase-polymerase chain reaction. Foodborne Pathog Dis. 2011 Apr;8(4):527-34. Epub 2011 Mar 7.
• Su X, MY Sangster, DSouza DH. 2011. Time-dependent effects of pomegranate juice and pomegranate polyphenols on foodborne viral reduction. Foodborne Pathog Dis. 2011 Nov; 8(11):1177-83. Epub 2011 Jul 21.
• Horm KM, DSouza DH. 2011. Survival of human norovirus surrogates in milk, orange, and pomegranate juice, and juice blends at refrigeration (4 degree C). Food Microbiol. 2011 Aug; 28(5):1054-61. Epub 2011 Mar 2.
• Radin D, DSouza DH. 2011. Evaluation of Two Primer Sets Using Newly Developed Internal Amplification Controls for Rapid Human Norovirus Detection by SYBR Green I Based Real-Time RT-PCR. Food Environ. Virol. 3 (2): 61-69. DOI: 10.1007/s12560-011-9057-6 Published: JUN 2011
• Techathuvanan C, Draughon FA, DSouza DH. 2011. Comparison of reverse transcriptase PCR, reverse transcriptase loop-mediated isothermal amplification, and culture-based assays for Salmonella detection from pork processing environments. J Food Prot. 2011 Feb;74(2):294-301.
• Miller ND, Davidson PM, DSouza DH. 2011. Real-time reverse-transcriptase PCR for Salmonella Typhimurium detection from lettuce and tomatoes. LWT-Food Sci Tech. 44 (4) 1088-1097. DOI: 10.1016/j.lwt.2010.08.003 Published: MAY 2011
• Su X, DSouza DH. 2011. Grape seed extract for control of human enteric viruses. Appl Environ Microbiol. 2011 Jun;77(12):3982-7. Epub 2011 Apr 15.
• Su X, DSouza DH. 2011. Trisodium phosphate for foodborne virus reduction on produce. Foodborne Pathog Dis. 2011 Jun;8(6):713-7. Epub 2011 Mar 7.

Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: The investigator's research efforts during this reporting period focused mainly on objectives 1, 2, and 3. (1) Novel rapid and robust molecular-based technologies for Salmonella detection were developed and applied to produce to enable tracking and transmission of outbreaks in the food environment. Sensitive real-time RT-PCR approaches based on mRNA were developed that are capable of detecting Salmonella from produce within 24 h. Methods to track outbreaks related to E. coli and Salmonella by pulse field gel electrophoresis (PFGE) enabled phylogenetic analysis of isolated strains. (2) Improved intervention strategies to control foodborne bacterial and viral pathogens in the environment and efficacy on food contact surfaces were also evaluated. Novel chemicals were used to determine inactivation of foodborne viral pathogens on food contact surfaces with improved efficacy. In collaboration with researchers at Rutgers State University of New Jersey, natural alternatives such as novel bioactive components of plants were also investigated for antiviral activity. Novel processing technologies to inactivate foodborne viral pathogens were carried out such as high pressure homogenization and high-intensity ultrasound in collaboration with researchers within FST-UT. (3) Fundamental transcriptomic-based approaches to determine appropriate control and mitigation strategies in food environments were also investigated. Gene expression profiling of E. coli O157:H7 under conditions of stress encountered in the food processing environments were determined using microarray analysis and RT-PCR approaches in collaboration with FST-UT faculty. PARTICIPANTS: Nathan Duray Miller, graduate student Katie Horm, graduate student Chayapa Techathuvanan, graduate student Xiaowei Su, Research Associate TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Robust, rapid, and sensitive detection methods for foodborne bacterial pathogens can help in the prevention of foodborne outbreaks related to produce and food environments. Rapid detection will help in the improvement of food safety and protect public health due to decreased foodborne disease outbreaks with decreased economical losses to agriculture. Novel natural ecofriendly intervention strategies that help to sustain the environment and simultaneously enhance safety of food products have potential applications in the food industry. Natural plant bioactive components that have nutritional benefits with added antimicrobial properties can be used as/in functional foods and beverages or as natural therapeutic agents. Ecofriendly, non-toxic, "green" approaches can be used as alternates to enhance produce safety and also protect the environment. Using fundamental "omics" approaches, the basis of foodborne bacterial survival and control in the food environment can be delineated to improve the overall safety of foods and the food environment.

Publications

• D. Radin, and D. H. DSouza. Rapid detection of human noroviruses in produce. 2nd Cost929 Symposium, Future Challenges in Food and Environmental Virology, Istanbul, Turkey, October, 2010.
• X. Su, and D.H. DSouza. Virucidal effects of TSP against foodborne viruses in produce. 2nd Cost929 Symposium, Future Challenges in Food and Environmental Virology, Istanbul, Turkey, October, 2010.
• Ganapathy, G., S. Zivanovic, and D. H. DSouza. In vitro time dependence effects of chitosan against foodborne viruses. UT-CEM Symposium, June, 2010. Oral presentation, Knoxville, TN
• K. Horm, F. Harte, and D. H. DSouza. Novel processing technologies for the inactivation of foodborne viral surrogates in milk and juice. Oral Presentaion, GAMMA SIGMA DELTA AWARD for outstanding research in agriculture. UT-CEM Symposium, June, 2010.
• X. Su, F. M. Harte, S. Zivanovic, D. H. DSouza. Effect of high-pressure homogenization and chitosan on the infectivity of human enteric viral surrogates. IFT Poster Presentation, Chicago, IL, July, 2010.
• R. H. Davis, D. H. DSouza, S. Zivanovic. The effect of different molecular weight chitosans on the inactivation of bacteriophages MS2 and phi X174. IFT Poster Presentation, Chicago, IL, July, 2010.
• D. Radin and D. H. DSouza. Developing Internal Amplification Controls to Compare and Improve Detection Sensitivity of Human Noroviruses from Clinical Samples by SYBR Green I Based Real-Time Reverse Transcription-PCR, ASM Poster, San Diego, CA, May 2010.
• Su, X., M. Y. Sangster, D. H. DSouza. Pomegranate juice and pomegranate polyphenols against the infectivity of human enteric viral surrogates. ASM Poster, San Diego, CA, May 2010.
• Su, X, A. Howell, D. H. DSouza. Time dependence effects of cranberry juice and cranberry PACs on the infectivity of human enteric viral surrogates. ASM Poster, San Diego, CA, May 2010.
• Su, X, A. Howell, D. H. DSouza. The effect of cranberry juice and cranberry PACs on the infectivity of human enteric viral surrogates. ASM KY-TN Regional Meeting, Knoxville, TN, October 17, 2009.
• Horm, K., F. Harte, X. Su, D. H. DSouza. High pressure homogenization for the inactivation of feline caliciviruses in orange juice and milk. ASM KY-TN Regional Meeting, Knoxville, TN, October 17, 2009.
• Techathuvanan, C., F. A. Draughon, and D.H. DSouza, Comparison of RT-PCR, RT-LAMP and cultural method for the detection of Salmonella from carcass rinse and carcass swabs. ASM KY-TN Regional Meeting, Knoxville, TN, October 17, 2009.
• Su, X., M.Y. Sangster, D. H. DSouza. 2010. In Vitro Effects of Pomegranate Juice and Pomegranate Polyphenols on Foodborne Viral Surrogates. Foodborne Pathogens and Disease Dec;7(12):1473-9.
• Su, X., A. B. Howell, D. H. DSouza. 2010. Antiviral effects of cranberry juice and cranberry proanthocyanidins on foodborne viral surrogates - A time dependence study in vitro. Food Microbiology 2010 Dec;27(8):985-91.
• Su, X, S. Zivanovic, D. H. DSouza. 2010. Inactivation of human enteric virus surrogates by high intensity ultrasound. Foodborne Pathogens and Disease Sep;7(9):1055-61.
• Su, X., Howell, A. B., D. H. DSouza. 2010. The effect of cranberry juice and cranberry proanthocyanidins on the infectivity of human enteric viral surrogates. Food Microbiology Jun;27(4):535-40. Epub 2010 Jan 25.
• Critzer F. J., D.H. DSouza, A.M. Saxton, D.A. Golden. 2010. Increased transcription of the phosphate-specific transport system of Escherichia coli O157:H7 after exposure to sodium benzoate. Journal of Food Protection May;73(5):819-24.
• DSouza, D. H., and X. Su. 2010. Efficacy of chemical treatments against murine norovirus, feline calicivirus and MS2 bacteriophage. Foodborne Pathogens and Disease Mar;7(3):319-26.
• Miller, N.D., F. A. Draughon, D. H. DSouza. 2010. Real-time Reverse-Transcriptase Polymerase Chain Reaction for Salmonella enterica detection from Jalapeno and Serrano Peppers. Foodborne Pathogens and Disease. Apr;7(4):367-73.
• Su, X., S. Zivanovic, D. H. DSouza. 2009. The effect of chitosan on the infectivity of murine norovirus, feline calicivirus and MS2 bacteriophage. Journal of Food Protection 72(12):2623-8.
• DSouza, D. H., X. Su, A. Roach, F. Harte. 2009. High pressure homogenization for the inactivation of human enteric virus surrogates. Journal of Food Protection Nov;72(11):2418-22.