Jordan Brad Webster
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Heat Resistance of Escherichia coli and Salmonella enterica in Ground Beef and Chicken by Jordan Brad Webster A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Food Science and Technology Department of Agricultural, Food and Nutritional Science University of Alberta © Jordan Brad Webster, 2018 Abstract Escherichia coli and Salmonella are contaminants in meat products and pose a risk for foodborne illness. Thermal lethality is the traditional method for controlling pathogens in meat products. It is recommended that ground beef and poultry products be cooked to internal temperatures of 71°C and 74°C, respectively. However, E. coli and Salmonella have a wide variance in resistance to heat. The aim of the current research was to determine the heat resistance of heat- resistant and heat-sensitive E. coli and Salmonella in ground beef and chicken and to determine the influence of the presence of a seasoning binder on heat resistance in meat products. Ground beef with and without 5.6% seasoning binder was inoculated and heated at temperatures between 55 and 70°C for different times. A linear regression model was used to calculate the D- and z- values. Seasoning binder increased the decimal reduction times (D-values) of heat-resistant E. coli AW1.7 but not of heat-resistant S. enterica ATCC 43845 in ground beef. To determine inactivation in patties, ground beef with and without 5.6% seasoning binder was inoculated with heat-resistant and heat-sensitive E. coli and Salmonella. When cooked to an internal temperature of 71°C, the presence of seasoning binder did not increase survival of heat-resistant or heat- sensitive E. coli and Salmonella in ground beef patties. Cooking ground beef patties to an internal temperature of 74°C achieved greater than 5-log (CFU/g) reduction of heat resistant E. coli AW1.7. The addition of 5.6% seasoning binder increased the survival of a cocktail of verotoxigenic E. coli inoculated in ground beef patties cooked to an internal temperature of 71°C. The cocktail was only reduced 2.8 log (CFU/g) in ground beef patties cooked to 71°C. To determine thermal inactivation in chicken nuggets, ground chicken was inoculated with heat- resistant and heat-sensitive E. coli and Salmonella, formed into chicken nuggets, heated and frozen. After being cooked to an internal temperature of 74°C cell counts of heat-resistant E. coli ii and Salmonella were reduced less than 3-log (CFU/g) and heat-sensitive E. coli were reduced by greater than 5-log (CFU/g). In conclusion, recommended cooking guidelines are not sufficient to prevent the survival of heat-resistant E. coli and Salmonella. The results of this study can be used by producers to help to assess their process and predict the survival of heat resistant organisms in ground beef and chicken products. iii Dedication To my life-advisor, my late grandfather Charles Daryl Webster: because I owe it all to you. Many Thanks! iv Acknowledgements Four years ago I was living in my car while working at a slaughterhouse and suffering from a parasite from a contaminated water source. Through some twist of fate and love of food science I continued forward torturing myself by pursing grad school. I would like to give a tremendous amount of thanks to my advisor Dr. Lynn McMullen for all her help over the years. She was my introduction and guide through the wonderful and fascinating world of food microbiology. She is always there to believe in me as a student and give me confidence as a person. Additionally, I would like to thank Dr. Michael Gänzle for always offering his much- appreciated critical input and reminder of the need for a sexy story. Thank you to my colleagues in 2-50 who were a constant source of help and encouragement. It was great sharing a laboratory with all of you during the last two years. Thank you to all those who came before me (some who were a part of the E. coli AW1.7 research tree) and who’s work furthered our understanding of heat resistance. A notable thank you to Brian Walker and Chandré Van De Merwe who started grilling the burgers that kicked off my research. I am grateful to my siblings and parents, who have provided me with moral and emotional support throughout my life. They showed me the comfort of a home and a hug. A very special gratitude to the funding source NSERC Create – Meat Education and Training Network who’s funding allowed me to develop skills and experience to be responsible in the meat industry. Cargill Inc. is acknowledged for supplying ground beef for this project. Finally thank you, potentially unknown reader, for taking the time to read my thesis. I hope my work, best summarized as science of fast food cooking, provides you with interest, insight, or at least entertainment in the fact that it is about burgers and chicken nuggets. v Table of Contents 1. Introduction and Literature Review ............................................................................................ 1 1.1 The meat industry ..................................................................................................................2 1.1.1 Overview of the Canadian beef industry ....................................................................... 2 1.1.2 Overview of the Canadian poultry industry ................................................................... 5 1.2 Organisms of concern ............................................................................................................6 1.2.1 Pathogenic Escherichia coli ........................................................................................... 6 1.2.2 Salmonella...................................................................................................................... 8 1.3 Thermal destruction of pathogens ........................................................................................10 1.3.1 Thermal processing ...................................................................................................... 10 1.3.2 Mechanisms of heat resistance..................................................................................... 13 1.3.3 Locus of heat resistance ............................................................................................... 15 1.3.4 Heat resistance in a food matrix ................................................................................... 17 1.3.5 Seasoning binder in food matrices ............................................................................... 18 1.4 Research objectives ..............................................................................................................20 2. Materials and Methods .............................................................................................................. 21 2.1 Bacterial strains and culture conditions ...............................................................................21 2.2 Determination of D- and z-values in ground beef ................................................................21 2.2.1 Ground beef preparation .............................................................................................. 21 2.1.2 Thermal Treatment....................................................................................................... 22 2.3 Heat treatment in ground beef..............................................................................................23 2.3.1 Burger preparation ....................................................................................................... 23 2.3.1.1 Effect of seasoning binder after 48 h storage ........................................................ 23 2.3.1.2 Effect of end point core temperatures ................................................................... 23 2.3.1.3 Effect of seasoning binder on survival of VTEC .................................................. 24 2.3.2 Cooking procedures and determination of survival in ground beef patties ................. 24 2.4 Heat treatment in ground chicken ........................................................................................26 2.4.1 Ground chicken preparation ......................................................................................... 26 2.4.2 Cooking procedures and determination of reduction in chicken nuggets .................... 26 2.5 Statistical analysis ................................................................................................................28 vi 2.5.1 D-, z-, and F-values ...................................................................................................... 28 2.5.2 Heat treatment in meat ................................................................................................. 30 3. Results ...................................................................................................................................... 31 3.1 Effect of seasoning binder on D- values of E. coli AW1.7 and S. enterica ATCC 43845 in ground beef ...........................................................................................................31 3.2 Effect of seasoning binder z-values of E. coli AW1.7 and S. enterica ATCC 43845 in ground beef ......................................................................................................................32 3.3 Survival of heat-resistant and heat-sensitive organisms in cooked ground beef patties ...................................................................................................................................33 3.3.1