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FACTORS AFFECTING GROWTH and CULTURING of Campylobacter Jejuni

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FACTORS AFFECTING GROWTH and CULTURING of Campylobacter Jejuni FACTORS AFFECTING GROWTH AND CULTURING OF Campylobacter jejuni By INDAUÊ IEDA GIRIBONI DE MELLO A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2002 COPYRIGHT 2002 by Indauê Ieda Giriboni de Mello I dedicate this dissertation to my mother, Vildes Claudio Giriboni de Camargo Mello, who has always believed in me and has provided me with unconditional love, guidance, encouragement, and financial support throughout my long and never ending education. The journey that culminated with this work may have never begun without my mother. ACKNOWLEDGMENTS First and foremost, I would like to express my most sincere and humble gratitude to God for giving me so many opportunities for personal and spiritual advancement, as well as the courage, perseverance, and determination to follow my dreams and goals. A dissertation such as this is the result of many individuals who have spent their time and efforts to assure that this work would be completed. I have many people to thank. I wish to thank my major professor, Dr. Sally Williams, for her assistance and support during my doctoral program. Thanks are also due to my supervisory committee members, Dr. Henry Aldrich, Dr. Gary Butcher, and Dr. Richard Miles, for their advice and suggestions during my doctoral program. Thank must be also granted to Dr. Ramon Littell for his help with the statistical analyses and to Dr. James Lindsay, who unknowingly sparked my interest and love for food microbiology. Finally, I thank the Department of Animal Science at the University of Florida for the financial support they have provided me with. My deepest appreciation goes to my greatest blessing, my fiancé, Luis Carlos Nogueira, the most decent, kind, loving and righteous person I have ever known. I am most thankful for his so–much–needed emotional support, iv encouragement, companionship, dedication and love during the most difficult times of my doctoral studies and life. I am grateful for his understanding and patience when I was mentally exhausted, for making me feel that everything would be fine, and above all else for reminding me to always have faith in God. Various others are appreciated for their indispensable and crucial help. A hearty and special thank you goes to Jan Hunt and Dr. Don Bark, from the U.S. Food and Drug Administration, for teaching me how to work with Campylobacter and for inspiring my career in food safety. A thank you is also dedicated to Donna Williams for her uncountable assistance with the electron microscope. Furthermore, I could not have hand–poured all the microbiological plates needed for my research without the unselfish aid of Sonia Nangó, Jennifer Gollwitzer and Dr. Clay Walker (and their automatic pour–plate machine), from the Oral Biology Department. Thanks are also extended to Anne Benner and Sergei Sennikov for helping me with the microbial and colorimetric analyses of the chicken breasts used in my study. Many other friends had a positive impact on my life and on this dissertation without even knowing. My dear friends Patrick Lannon, Andreas Kavazis, Justin Sobota, Bernhard Bodmann, and Dr. Ronald Schmidt believed in me when they decided to help me create the International Gourmet Association (IGA). The IGA meetings became my safe harbor where I was able to be myself, learn about leadership, patience, tolerance, sagacity, and where I had the opportunity to meet several individuals who became important in some aspects v of my life. Many thanks go also to all my friends who attended to the exit seminar of my defense and gave me so much support, cheering me up and keeping me confident. A special thank you is extended to a few very dear friends, Dr. Maria Luiza Braga Fernandes, Dr. Rogério Lacaz Ruiz, and Gillian Folkes, for always being there for me when I needed something, no matter how far or close I might be. Their friendship will be forever cherished. A gesture of appreciation goes to Dr. Anthony Pescatore and Mike Ford, who have always kept an interest in my academic progress even after I left the University of Kentucky and for taking me under their wings in every scientific meeting we attended. Their warm thoughts and inquiries were always received with happiness. And above all else, I would like to express my most sincere gratitude to a special friend, Luis Maia de Mello Massa, for assisting me through the admission process of my doctoral program and for his encouragement, especially during the first two years of my PhD work. I have been blessed in my life with a remarkably supportive family. This dissertation also belongs to them for encouraging my continuous pursuit of academic excellence and for supporting me in every step along the way. Many thanks go to my beloved sister, Amaitê Iara Giriboni de Mello, and my dearest brother, Rubner Vilens Giriboni de Mello, for the love they have always given me. My sister has been an inspiration and an example of dedication to professional life; her vehement sense of justice has constantly reminded me that vi fairness and moral rightness must prevail, at any cost. And foremost, I thank her for providing me with an enormous demonstration of her love during my doctoral studies by naming her daughter after me. In spite of my brother’s desire to pass unnoticed through my life, I know that hidden inside him, within his big heart, is a genuine wish to see me succeed and thrive; and for that I am most grateful. In all the ups and downs that I have encountered, he has always kept the doors open for me. I admire his search for self–identity and truth. A profound gratefulness, respect and admiration are devoted to my mother, Vildes Claudio Giriboni de Camargo Mello, to whom this work is specially dedicated. And last but not least, I deeply thank my late father, Rubens Camargo Mello, who always taught me the value and importance of education, perseverance, and endless quest for knowledge. Of humble background, he succeeded and worked hard throughout his life to provide for his family even after his death. For this and so many other reasons I am honored to be his daughter. vii TABLE OF CONTENTS page ACKNOWLEDGMENTS ............................................................................................... iv LIST OF TABLES............................................................................................................. xi LIST OF FIGURES.........................................................................................................xiv ABSTRACT ...................................................................................................................xvii CHAPTER 1 INTRODUCTION .......................................................................................................1 2 LITERATURE REVIEW..............................................................................................4 General Microbiology of Campylobacter spp. .......................................................... 4 Historical Background.......................................................................................... 4 Morphological and Biochemical Properties .................................................... 11 Antibiotic Resistance and Sensitivity ............................................................... 16 Pathogenesis and Epizootiology....................................................................... 18 Virulence and Toxin Production....................................................................... 21 Poultry as Reservoir and Vehicle of Campylobacter............................................. 29 General Properties of Fresh Poultry ................................................................. 32 Poultry microorganisms...............................................................................33 Early studies on Campylobacter and poultry..............................................37 Prevention and Control of Campylobacter ........................................................ 45 Factors that Have a Potential Effect on Growth and Survival of Campylobacter ............................................................................................................ 50 Allyl Isothiocyanates .......................................................................................... 50 Acid–Shock........................................................................................................... 59 Importance of Acid–shock in Foodborne Pathogens..................................... 62 Methods for Culturing and Analyzing Campylobacter........................................ 68 Selective Culture Media ..................................................................................... 68 Culturing Methodology ..................................................................................... 76 Porcine bile juice in culture media .............................................................78 Porcine gastric mucin in culture media .....................................................81 Microbiological Electron Microscopy............................................................... 85 viii 3 ANTIMICROBIAL PROPERTIES OF ALLYL ISOTHIOCYANATE.................93 Material and Methods ............................................................................................. 94 Selective Culture Media and Agar.................................................................... 94 Bacterial Culture.................................................................................................
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