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The Efficacy of Antimicrobials for the Control of Alicyclobacillus Acidoterrestris in Fruit and Vegetable Juices by Angela D. Ha

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The Efficacy of Antimicrobials for the Control of Alicyclobacillus Acidoterrestris in Fruit and Vegetable Juices by Angela D. Ha The Efficacy of Antimicrobials for the Control of Alicyclobacillus acidoterrestris in Fruit and Vegetable Juices By Angela D. Hartman A Thesis submitted in partial fulfillment of the Requirements for the degree of Master of Science in Food Science and Technology Virginia Polytechnic Institute and State University, Blacksburg, Virginia Approved: _________________________ Robert C. Williams, Chair _____________________ ______________________ Susan S. Sumner Bruce W. Zoecklein June 12, 2003 Blacksburg, Virginia Keywords: Alicyclobacillus acidoterrestris, juices, antimicrobials Copyright 2003, Angela Hartman The Efficacy of Antimicrobials for the Control of Alicyclobacillus acidoterrestris in Fruit and Vegetable Juices Angela D. Hartman (ABSTRACT) The efficacy of antimicrobials for control of A. acidoterrestris spoilage in juices was analyzed. Apple and tomato juices were inoculated with 4 log spores/ml. Antimicrobials were added at: 1000, 500 and 250 ppm (sodium benzoate, potassium sorbate, and sodium metabisulfite); 500, 250, and 125 ppm (cinnamic acid, dimethyl dicarbonate, and ascorbic acid); 125, 75 and 25 ppm (lysozyme); and 5, 3, and 1 IU/ml (nisin). In apple juice, A. acidoterrestris population reductions were caused by the following antimicrobials (reduction in log CFU/ml): lysozyme - all levels and nisin - 5 IU/ml (5.1), nisin - 3 IU/ml (4.2), cinnamic acid - 125 ppm (3.1), cinnamic acid - 250 ppm (2.6), potassium sorbate - 250ppm (2.5), nisin - 1 IU/ml (2.4), potassium sorbate - 500 and 1,000 ppm (2.3), dimethyl dicarbonate - 500 ppm (1.9), cinnamic acid - 500 ppm (1.4). In tomato juice, A. acidoterrestris population reductions were caused by the following antimicrobials (reduction log CFU/ml): nisin - all levels and lysozyme - 125 ppm and 75 ppm (4.4), lysozyme - 25 ppm (3.8), potassium sorbate - 500 ppm (2.6), cinnamic acid - 500 ppm (2.5), cinnamic acid - 250 ppm (2.4), cinnamic acid - 125 ppm (2.1), potassium sorbate - 1,000 ppm (1.9), and potassium sorbate - 250 ppm (1.6). Antimicrobial treatments: nisin - ≥ 1 IU/ml, lysozyme - ≥ 25 ppm, cinnamic acid - ≥ 125 ppm, and potassium sorbate - ≥ 250 ppm may be appropriate controls to prevent A. acidoterrestris spoilage in juices or juice containing beverages. ACKNOWLEDGMENTS I would like to think my graduate advisor Dr. Robert Williams for all of his help and knowledge over the span of my research so that my project could be at its full potential. I would also like to thank him for taking a chance on a student he didn’t know and for allowing me to ask numerous questions and take risks. His encouragement and advice always kept me motivated so that my project ideas could be fulfilled. I am excited to continue to work with him on my PhD. I would also like to thank my committee members, Dr. Susan Sumner and Dr. Bruce Zoecklein. Dr. Sumner really allowed me to do a project that was different and fit my style. I would also like to thank my family for all of the love, encouragement, and advice along the way. I want to thank Ricardo Richardson and Susan Horwatt for all of their support and laughter to ensure that all of my goals and dreams were met. I would also like to thank Richard Smith for all of his insight on my research, for always giving me ideas that could be employed in my research, and for giving me an industry standpoint to the problem of A. acidoterrestris- associated spoilage. A big thanks to undergraduate researchers Katie Pribisko and Zelmira Arias for all of their help in the lab and on other experiments along the way. Thank you so much to all of the staff that have helped me, including Brian Smith for his knowledge about many different things and Brian Yaun for helping me at a moments notice. Joe Boling, Harriet Williams, Walter Hartman and Hengjian Wang have also been very helpful in helping complete this project. I would also like to thank my statistician, Wes Schilling for the hours helping me with my statistics and the numerous questions that went along with it. I would like to thank all of my fellow graduate students for allowing me to not be totally consumed in only research. Thank you especially to Renee Raiden, Megan Hereford, Karol Gailunas, Donna Greene, Jomel Quicho, Christine Piotrowski, and Gabe Sanglay. In addition, I would like to think Canadian Inovatech for the contribution of the lysozyme and all of their help. Finally I would like to thank God for giving me this great opportunity in life and blessing me each day with great things and life lessons. iii TABLE OF CONTENTS page Acknowledgements..................................................................................................................... iii Table of Contents.........................................................................................................................iv List of Tables ............................................................................................................................. vii List of Figures........................................................................................................................... viii Introduction/Justification ..............................................................................................................1 CHAPTER 1: Literature Review.................................................................................................4 A. Alicyclobacillus spp. ................................................................................................................4 B. Alicyclobacillus acidoterrestris................................................................................................6 1. Ecology .........................................................................................................................6 2. Heat and Acid Tolerance ..............................................................................................6 3. Biochemistry of Heat and Acid Tolerance....................................................................8 4. Enumeration and Identification Techniques .................................................................9 5. Sources........................................................................................................................11 6. Spoilage.......................................................................................................................16 7. Spoilage Compounds ..................................................................................................19 8. Control Measures........................................................................................................20 C. Antimicrobials........................................................................................................................24 D. Nisin 1. Chemical Description and Approval...........................................................................24 2. Mechanism of Action..................................................................................................25 3. Interaction with Juice Components.............................................................................27 4. Previous Work............................................................................................................29 E. Sodium Benzoate 1. Chemical Description and Approval...........................................................................30 2. Mechanism of Action..................................................................................................31 3. Interaction with Juice Components.............................................................................32 4. Previous Work............................................................................................................33 F. Potassium Sorbate 1. Chemical Description and Approval...........................................................................33 2. Mechanism of Action..................................................................................................34 3. Interaction with Juice Components.............................................................................37 4. Previous Work............................................................................................................39 G. Trans-Cinnamic Acid 1. Chemical Description and Approval...........................................................................39 2. Mechanism of Action..................................................................................................40 iv 3. Interaction with Juice Components.............................................................................41 4. Previous Work............................................................................................................41 H. Sodium Metabisulfite 1. Chemical Description and Approval...........................................................................42 2. Mechanism of Action..................................................................................................43 3. Interaction with Juice Components.............................................................................44 4. Previous Work............................................................................................................45 I. Lysozyme 1. Chemical Description and Approval...........................................................................46 2. Mechanism of Action..................................................................................................47 3. Interaction with Juice Components.............................................................................48
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