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Bovine Digital Dermatitis: Natural Lesion Development and Experimental Induction Adam C Iowa State University Capstones, Theses and Graduate Theses and Dissertations Dissertations 2015 Bovine digital dermatitis: natural lesion development and experimental induction Adam C. Krull Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/etd Part of the Animal Sciences Commons, Microbiology Commons, and the Veterinary Medicine Commons Recommended Citation Krull, Adam C., "Bovine digital dermatitis: natural lesion development and experimental induction" (2015). Graduate Theses and Dissertations. 14540. https://lib.dr.iastate.edu/etd/14540 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Graduate Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Bovine Digital Dermatitis: Natural lesion development and experimental induction by Adam C. Krull A dissertation submitted to the graduate faculty in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Major: Veterinary Microbiology Program of Study Committee: Paul J. Plummer, Major Professor Gregory J. Phillips Jan K. Shearer Patrick J. Gorden Grant A. Dewell Iowa State University Ames, Iowa 2015 Copyright © Adam C. Krull, 2015. All rights reserved. ii TABLE OF CONTENTS Page ACKNOWLEDGMENTS ..................................................................................... iv ABSTRACT .......................................................................................................... v CHAPTER 1 GENERAL INTRODUCTION .................................................... 1 Introduction .................................................................................................... 1 Thesis Organization ......................................................................................... 2 Literature Review ............................................................................................ 3 Prevalence and Economic Impact of Digital Dermatitis .............................. 4 DD Morphology – Macroscopic and Microscopic ...................................... 5 Etiology of Digital Dermatitis .................................................................... 7 Treatment of Digital Dermatitis ................................................................. 22 Prevention of Digital Dermatitis ................................................................ 23 Significance of Research ................................................................................. 24 References .................................................................................................... 24 CHAPTER 2 DIGITAL DERMATITIS: NATURAL LESION PROGRESSION AND REGRESSION IN HOLSTEIN CATTLE OVER THREE YEARS .............. 32 Abstract .................................................................................................... 33 Introduction .................................................................................................... 34 Materials and Methods .................................................................................... 36 Results .................................................................................................... 40 Discussion .................................................................................................... 53 Acknowledgments ........................................................................................... 60 References .................................................................................................... 60 CHAPTER 3 DEEP SEQUENCING ANALYSIS REVEALS THE TEMPORAL MICOROBIOTA CHANGES ASSOSIATED WITH THE DEVELOPMENT OF BOVINE DIGITAL DERMATITIS ...................................................................... 63 Abstract .................................................................................................... 64 Background .................................................................................................... 65 Materials and Methods .................................................................................... 67 Results .................................................................................................... 75 Discussion .................................................................................................... 92 Acknowledgments ........................................................................................... 100 References .................................................................................................... 100 iii Appendix A .................................................................................................... 106 CHAPTER 4 A HIGHLY EFFECTIVE PROTOCOL FOR THE RAPID AND CONSISTENT INDUCTION OF DIGITAL DERMATITIS IN HOLSTEIN CALVES .................................................................................................... 123 Abstract .................................................................................................... 124 Introduction .................................................................................................... 124 Materials and Methods .................................................................................... 128 Results .................................................................................................... 135 Discussion .................................................................................................... 146 Acknowledgments ........................................................................................... 158 References .................................................................................................... 158 Appendix B .................................................................................................... 162 CHAPTER 5 CONCLUSIONS AND FUTURE RESEARCH........................... 170 Summary and Conclusions .............................................................................. 170 Recommendations for Future Research ............................................................ 172 References ..................................................................................................... 174 iv ACKNOWLEDGMENTS I would like to thank my committee chair, Paul Plummer, and my committee members, Gregory Phillips, Jan Shearer, Patrick Gorden, and Grant Dewell for their guidance and support throughout the course of this research. In addition, I would also like to thank my friends, colleagues, the department faculty and staff for their encouragement and support over the last five years. A special thanks to my wife and colleague, Amanda Kreuder-Krull, for her support and recommendations in all aspects of trial design, implementation, and interpretation of results. I want to also offer my appreciation all of the student’s that were involved in these projects, especially Cory Haglund, Michael Slattery, Tim Kruse, Lance Dahlquist, Mackenzie Dickson Chris Rock, and Laura Bro. Thanks also to the technical support staff of the Iowa State Veterinary Diagnostic Laboratory histopathology laboratory as well as to Dr. Gregory Stevenson for the microscopic photography. I would also like to thank the ISU Dairy Farm, ISU LIDIF, and ARS staff for their assistance in sample collection and animal management. Funds for this research project were provided by Boehringer Ingelheim Vetmedica Inc. and Iowa State University startup funds. v ABSTRACT Bovine Digital Dermatitis (DD) is a leading cause of lameness in dairy cattle in the US with more than 70% of herds affected. Despite 40 years of research, the definitive etiologic agent(s) associated with the disease process is unknown. While clinical lesions have been well described, little is known about the macroscopic, microscopic, and bacterial changes associated with the early stages of lesion development from normal skin to clinical lesions. The goal of this dissertation was to describe the temporal changes associated with lesion development in Holstein dairy cattle, particularly early stage lesions, and develop a model for lesion induction. By following a cohort of Holstein dairy cows for a three year period, several important epidemiologic findings were recognized. In the absence of control measures, DD lesions developed at a rate of 4 lesions per 100 cow feet-months, with the average time for a lesion to develop being 133 days. From the recognition of the macroscopic changes that preceded clinical DD lesions, a novel scoring system was developed. While 20% of the feet observations had clinical DD lesions, an additional 55% of observations had lesions that were indicative of early DD lesion development. Biopsies from these different stages of lesion development were submitted for metagenomic analysis using next generation sequencing. The bacterial microbiota of these biopsies was found to progress through a systematic series of changes that correlate with the macroscopic lesion scoring system with the microbiota of each stage being statistically different from other stages. As one of the major goals for these studies was to gain a better understanding of the etiology of disease, an experimental model was needed to test candidate pathogens. Four preliminary studies were conducted to optimize conditions and methodologies for induction of DD lesions that led to a vi final consensus protocol that was able to induce
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