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Combination of Ultra-High Pressure and Xanthene-Derivatives to Inactivate Food-Borne Spoilage and Pathogenic Bacteria

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Combination of Ultra-High Pressure and Xanthene-Derivatives to Inactivate Food-Borne Spoilage and Pathogenic Bacteria COMBINATION OF ULTRA-HIGH PRESSURE AND XANTHENE-DERIVATIVES TO INACTIVATE FOOD-BORNE SPOILAGE AND PATHOGENIC BACTERIA DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Joy Gail Waite, M.S. * * * * * The Ohio State University 2007 Dissertation Committee: Approved by Professor Ahmed E. Yousef, Adviser Professor Steven J. Schwartz _________________________________ Professor MacDonald Wick Adviser Food Science and Nutrition Graduate Program Professor Jeff Culbertson ABSTRACT Food processing methods can lead to the development of processing-resistant spoilage and pathogenic microorganisms that can cause further cost to processors and increased health risks to consumers. Thus, inactivation of these potentially problematic bacterial strains by combinations of physical and/or chemical treatments needs to be investigated. Previous studies have shown combination treatments of ultra-high pressure (UHP) with antimicrobial peptides, oxidants, and antioxidants to be efficacious at inactivating bacteria. The current study investigates the use of UHP in combination with hydroxyxanthenes, including FD&C Red No. 3, to inactivate pressure-resistant strains of Lactobacillus plantarum, Listeria monocytogenes, and Escherichia coli O157:H7. Combination treatments of UHP (400 MPa, 3 minutes) and FD&C Red No. 3 (3 to 10 ppm) resulted in significant enhancement of inactivation of several pressure-resistant Gram-positive and Gram-negative strains. FD&C Red No. 3 is a known photosensitizer and was capable of inactivating Gram-positive species with short exposure to ambient light (15-30 minutes); however this compound had no effect on Gram-negative species without UHP treatment. UHP was found to cause an irreversible change in the barrier properties of the outer membrane with pressure treatments above 250 MPa leading to accumulation of FD&C Red No. 3 correlating with cell inactivation. Inactivation of Gram-positive and Gram-negative species by combination treatment was light-dependent ii with low UHP treatments (<400 MPa), indicating a role of photooxidation. However, with increasing pressures (>400 MPa) a light-independent mechanism was identified for all species tested. Efficacy of light-dependent and light-independent inactivation was determined in the food systems: carrot juice and turkey meat product. Combination treatment was effective against L. monocytogenes and E. coli O157:H7 in carrot juice with FD&C Red No. 3 at concentrations of 10 to 100 ppm. These strains were resistant to inactivation by combination treatment in turkey meat product, indicating that specific food components or composition may decrease the efficacy of these treatments. Further studies were completed to determine the impact of superoxide production on the inactivation of E. coli K12 wild-type and sod mutants. Mutant strains were not significantly different in sensitivity to any of the treatments tested, however, all E. coli K12 strains were significantly more sensitive to UHP and UHP-FD&C Red No. 3 combination treatments when treated and recovered under aerobic conditions compared to anaerobic conditions. Pressure-resistant strains were treated with UHP alone and in combination with FD&C Red No. 3 under aerobic and anaerobic conditions to determine the impact of type I and type II photooxidation on inactivation. The majority of inactivation by combination treatment with light exposure is due to type I photooxidation based on little difference between aerobic and anaerobic treatments. Interestingly, inactivation without light exposure was oxygen dependent for all microorganisms tested. Further study is needed to determine if oxygen dependent effect is due to oxygen presence during treatment or during recovery. Inactivation of UHP processing-resistant microorganism can be achieved using combination treatment of UHP and FD&C Red No. 3 in food systems. iii ACKNOWLEDGMENTS I would like to take this opportunity to thank the people that have made this research and my education at The Ohio State University possible. First and foremost, I would like to thank Dr. Ahmed Yousef for sharing his unbelievable wisdom and passion for research and his unwavering trust in my abilities. Your support has allowed me to grow into a better researcher and teacher; I only hope to possess a small fraction of your mentoring abilities and compassion for others. Secondly, I would like to thank the members of the Yousef lab for their friendship, encouragement, and wisdom: Yoon- Kyung Chung, Zengguo (France) He, Luis Rodriguez-Romo, Mohammed Khadre, Aaron Malone, Erin Horton, Mustafa Vurma, Shara Johnson, Xueying Zhang, Yuan Yan, Jennifer Perry, Amrish Chawla, and Joe Jones. I would like to specifically thank Joe Jones for being a wonderful person and friend and having an extraordinary ability to learn from someone as disorganized as myself. Thank you, Joe, for making dilution blanks and liters upon liters of media for me; but above all for all of the hard work and time spent on the salad dressing project, you are a life-saver, or more appropriately a project-saver. I would like to thank my committee members, Dr. Steven Schwartz, Dr. MacDonald Wick, and Dr. Jeff Culbertson for their patience through the latter stages of iv this process. Thank you Dr. Schwartz and Dr. Wick for teaching graduate courses that were thought-provoking and educational. Thank you to Dr. Culbertson for listening to my crazy rants about teaching problems. A special thanks to James Douglass, Richard Nist, Dan Aruscavage, and Michele Manuzon for all of the help with FST/MICRO 636.02, putting up with my rantings, and laughing together over “maximum penetration number” and “Asperilla”. Extra special thanks to Matt Mezydlo for making the term projects work; they would have failed without you. I would especially like to thank the extremely close friends I have made over the last few years: Mary Kay Folk, Jennifer Ahn-Jarvis, April Wax, Julie Jenkins, Nurdan Kocaoglu-Vurma and Robert King. You have made the time go by quickly and have helped distract me when necessary for me to retain my sanity. Special thanks to my mother Sharon, my father Joe, and my sisters Laurie and Mindy for believing in me and supporting my crazy ideas. Most importantly, I would like to thank my son Allan. You are the best kid in the world, thank you for being a wonderful, caring, and unique individual; I love you more than anything. v VITA June 20, 1980………………………………………. Born, Florence, Oregon, U.S.A. 2002……………………........................................... B.S. Food Science and Technology, Oregon State University. 2004………………………………………………... M.S. Microbiology, Oregon State University 2004-present……………………………………….. Graduate Research and Teaching Associate, The Ohio State University PUBLICATIONS Research Publication 1. Waite, J.G., M.A. Daeschel. 2007. Contribution of wine components to inactivation of food-borne pathogens. J. Food Sci. 72(7):M286-M291. FIELDS OF STUDY Major Field: Food Science and Nutrition vi TABLE OF CONTENTS Page Abstract................................................................................................................................ ii Acknowledgements..............................................................................................................iv Vita.......................................................................................................................................vi List of Tables .......................................................................................................................ix List of Figures......................................................................................................................xi Chapters: 1. Literature Review...........................................................................................................1 1.1. Color and food ........................................................................................................1 1.2. Xanthene dyes.......................................................................................................23 1.3. Photosensitizers and photooxidation ...................................................................36 1.4. Ultra-high pressure ...............................................................................................61 1.5. “Hurdle” Technologies .........................................................................................75 2. Xanthene-derived food colorants sensitize processing-resistant foodborne pathogenic and spoilage bacteria to ultra-high pressure.................................................................96 3. Destabilization of the outer membrane of Escherichia coli by physical and chemical methods leads to inactivation by FD&C Red No. 3 ..................................................119 4. Contribution of photooxidation to inactivation of Gram-positive and Gram-negative bacteria by ultra-high pressure and xanthene-derivatives..........................................145 5. Inactivation of bacteria by ultra-high pressure and xanthene-derivatives: a secondary light-independent mechanisms...................................................................................166 6. Efficacy of FD&C Red No. 3 and ultra-high pressure combination treatment against foodborne pathogens in food systems........................................................................168 7. Impact of aerobic and anaerobic treatment and recovery on inactivation
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