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An Environmental Approach to Food Safety Assessment Using Artisan Cheese and Fresh Produce As Model Systems Marie Limoges University of Vermont

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An Environmental Approach to Food Safety Assessment Using Artisan Cheese and Fresh Produce As Model Systems Marie Limoges University of Vermont University of Vermont ScholarWorks @ UVM Graduate College Dissertations and Theses Dissertations and Theses 2019 An Environmental Approach To Food Safety Assessment Using Artisan Cheese And Fresh Produce As Model Systems Marie Limoges University of Vermont Follow this and additional works at: https://scholarworks.uvm.edu/graddis Part of the Agriculture Commons, Food Science Commons, and the Plant Sciences Commons Recommended Citation Limoges, Marie, "An Environmental Approach To Food Safety Assessment Using Artisan Cheese And Fresh Produce As Model Systems" (2019). Graduate College Dissertations and Theses. 1013. https://scholarworks.uvm.edu/graddis/1013 This Dissertation is brought to you for free and open access by the Dissertations and Theses at ScholarWorks @ UVM. It has been accepted for inclusion in Graduate College Dissertations and Theses by an authorized administrator of ScholarWorks @ UVM. For more information, please contact [email protected]. AN ENVIRONMENTAL APPROACH TO FOOD SAFETY ASSESSMENT USING ARTISAN CHEESE AND FRESH PRODUCE AS MODEL SYSTEMS A Dissertation Presented by Marie A. Limoges to The Faculty of the Graduate College of The University of Vermont In Partial Fulfillment of the Requirements For the Degree of Doctor of Philosophy Specializing in Animal, Nutrition and Food Sciences January, 2019 Defense Date: November 13 th , 2018 Dissertation Examination Committee: Catherine W. Donnelly, Ph.D., Advisor Deborah A. Neher, Ph.D., Chairperson Omar A. Oyarzabal, Ph.D. Paul S. Kindstedt, Ph.D. Cynthia J. Forehand, Ph.D., Dean of the Graduate College Abstract This dissertation examined recently issued regulatory standards using cheese and produce as model systems. FDA’s 2015 Domestic and Imported Cheese and Cheese Products Compliance Program Guidelines (CPG) E. coli standards on cheese safety, and the extent to which these standards affect domestic and imported cheese commerce, was assessed. Results from FDA’s Domestic and Imported Cheese Compliance Program for samples collected between January 1, 2004 and December 31, 2006 were analyzed. Of 3,007 cheese samples tested for non-toxigenic E. coli, 76% (2,300) of samples exceeded 10/g, FDA’s target for regulatory activity. In cheese samples containing E. coli levels of 10/g and 100/g, there was no statistically significant association with presence of Listeria monocytogenes . However, associations between Staphylococcus aureus levels of 10,000 CFU/g and presence of Salmonella and L. monocytogenes were statistically significant, indicating that EU regulations targeting S. aureus as the pathogen of concern may be more appropriate than E. coli for cheese safety assessment. Compost amended soils in the Northeastern U.S. were assessed for the presence and survival of E. coli and Listeria spp. against FDA Food Safety Modernization Act (FSMA) requirements. Manure and poultry litter-based biological soil amendments of animal origin (BSAAO) must achieve pathogen reduction to reduce risk of pathogen contamination on the harvested produce. Two trials of replicated field plots of loamy (L) or sandy (S) soils were tilled and un-amended (NC) or amended with dairy compost (DC), poultry litter compost (PL), or poultry pellets (PP). Colony count and most probable number (MPN) methods were used to determine persistence of E. coli in these plots over 104 days post-inoculation. Detection of indigenous Listeria spp. were also examined in all plots. Higher E. coli populations were observed in PL and PP amended soils when compared to DC and NC plots. E. coli was detected at low levels on radish crops, where PL treatments encouraged greater levels of survival and growth than DC or NC. Study results verify that a 120 day interval following BSAAO application should be sufficient to ensure food safety of edible crops subsequently planted on these soils. The sensitivity of environmental monitoring methods and collection formats were evaluated to identify optimal procedures for detection of Listeria spp. on product contact surfaces within artisan cheese production environments. Four environmental surfaces (dairy brick, stainless steel, plastic, and wood; n=405/surface type) were inoculated with L. innocua , L.m. ATTC ® 19115 and L.m. 1042, at high (10 6-10 7/cm 2) and low (0.1- 1/cm 2) target concentrations. Inoculated surfaces were swabbed with World Bioproducts © EZ Reach TM and 3M TM environmental swabs (3M TM ). Five enrichment and enumeration methods were used to compare sensitivity of recovery between environmental swabs. All swab formats performed equally on all environmental surfaces at high target concentrations. At low concentrations, Petrifilm TM and WBEZ swabs recovered Listeria spp. from 87.5% of plastic, stainless steel, and dairy brick surfaces, but only 62.7% of wooden surfaces; recovering 14.8%, 77%, and 96.3% of cells from initial inoculations of 0.01, 0.1, and 1/cm 2, respectively . Our data demonstrate that results may be discrepant due to variation in the porosity of environmental surfaces and should be taken into consideration when implementing environmental sampling plans. Results from this thesis can be used to inform regulatory policy and help to achieve improved food safety. Acknowledgments After being involved with the Nutrition and Food Sciences community at the University of Vermont throughout my undergraduate and Master of Science in Dietetics graduate studies, and now my doctorate degree, I feel extremely fortunate to have connected and worked with all faculty, staff, and students. I especially want to thank Lacey Ure, Ann Martin, and Dr. Jean Harvey for their support and assistance during this time. While this dissertation was an independent work, I want to express my deepest gratitude to my advisor, Dr. Catherine Donnelly for her direction, encouragement, and impressive mentorship. I appreciate her kindness and willingness to assist with challenges while simultaneously fostering my independent analytical and critical thinking skills. I also wish to give thanks to the Chair of my committee, Dr. Deborah Neher, who has been a remarkable mentor, given a great amount of support, and has further cultivated my interests in mathematics and statistics. She has been especially instrumental during her involvement in the compost amended soils proJect. I am very fortunate to have such a wonderful doctorate committee for which I would also like to thank Dr. Omar Oyarzybal and Dr. Paul Kinstedt for their time and contributed expertise to my dissertation. I would like to give a special thanks to Tom Weicht for his assistance and tremendous support throughout the compost amended soils proJect and a big thank you to Dr. Rachel Schattman for her collaboration and help during the second trial of the compost amended soils proJect. I also wish to thank Dr. Manan Sharma and Dr. Patricia Millner of the USDA Agriculture Research Service for their technical support and guidance during my studies. I want to thank Alan Howard for his guidance with statistical analyses and consistent availability for questions. I wish to thank all other graduate students for their ii help and humor and I especially want to thank Dr. Panos Lekkas for his mentorship and training when I first started the program. I also would like to thank my family and friends for their words of encouragement and understanding, as I have had to dedicate much of my time to this degree, resulting in missed events over the years. Lastly but ever so importantly, mom and dad, I cannot thank you both enough for your ongoing support, unconditional love, and words of wisdom. Your positivity and faith in me throughout this process has consistently redeemed my motivation to achieve and complete this lifelong goal. iii Table of Contents ABSTRACT ........................................................................................................................ I ACKNOWLEDGMENTS ............................................................................................... II LIST OF TABLES .......................................................................................................... IX LIST OF FIGURES ......................................................................................................... X CHAPTER 1: COMPREHENSIVE LITERATURE REVIEW ................................... 1 Pathogens of Concern ....................................................................................................... 1 Listeria monocytogenes .................................................................................................. 1 Genus Listeria ........................................................................................................................................ 1 Virulence ............................................................................................................................................... 2 Enrichment Methods .............................................................................................................................. 5 Dual Enrichment .................................................................................................................................... 8 Agar Plating Media ................................................................................................................................ 9 Polymerase Chain Reaction (PCR) ...................................................................................................... 11 Molecular Subtyping...........................................................................................................................
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