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Evaluation of Laser and LED Phototherapy for the Treatment of Canine Acral Lick Dermatitis and Staphylococcus Pseudintermedius in Vitro

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Evaluation of Laser and LED Phototherapy for the Treatment of Canine Acral Lick Dermatitis and Staphylococcus Pseudintermedius in Vitro Evaluation of laser and LED phototherapy for the treatment of canine acral lick dermatitis and Staphylococcus pseudintermedius in vitro THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By Amy H. Schnedeker, D.V.M. Graduate Program in Comparative and Veterinary Medicine The Ohio State University 2017 Master’s Examination Committee: Dr. Lynette Cole, Advisor Dr. Sandra Diaz Dr. Gwendolen Lorch Dr. Joshua Daniels Dr. Paivi Rajala-Schultz i Copyright by Amy H. Schnedeker 2017 i Abstract Staphylococcus pseudintermedius is the most common cause of bacterial skin infections in dogs. Methicillin-resistant infections have become more common and are challenging to treat. Blue light phototherapy may be an option for treating these infections. The objective of this study was to measure the in vitro bactericidal activity of 465-nm blue light on methicillin-susceptible Staphylococcus pseudintermedius (MSSP) and methicillin-resistant Staphylococcus pseudintermedius (MRSP). We hypothesized that irradiation with blue light would kill MSSP and MRSP in a dose-dependent fashion in vitro as previously reported for methicillin-resistant Staphylococcus aureus (MRSA). In six replicate experiments, each strain (MSSP: n=1), (MRSP ST-71 [KM1381]: n=1) and (MRSA [BAA-1680]: n=1) were cultivated on semisolid media, irradiated using a 465-nm blue light phototherapeutic device at the following cumulative doses: 56.25, 112.5, and 225 J/cm2 and incubated overnight at 35oC. Controls were not irradiated. Colony counts (CC) were manually performed. Descriptive statistics were performed and treatment effects assessed using the Mann-Whitney-Wilcoxon rank-sum test. Bonferroni- corrected rank sum tests were performed for post-hoc analysis when significant differences were identified. ii There was a significant decrease in CC with blue light irradiation at all doses for MRSA (P=0.0006) but not for MSSP (P=0.131) or MRSP (P=0.589). Blue light phototherapy significantly reduced CC of MRSA, but not of MSSP or MRSP. The mechanism for the relative photosensitivity of the MRSA isolate is unknown, but is hypothesized to be due to an increased concentration of porphyrin in S. aureus relative to S. pseudintermedius, which would modulate blue light absorption. Canine acral lick dermatitis (ALD) is a frustrating, chronic disease that can have numerous primary causes including pruritic and painful diseases. Conventional therapy generally consists of systemic antibiotics and anti-anxiety medications. Low-level laser therapy (LLLT) is an alternative therapy that has been used to treat conditions associated with inflammation and pain. The objective of this study was to determine whether the use of LLLT combined with conventional therapy would result in a significant decrease in the licking of the ALD lesion than conventional therapy alone. We hypothesized that the combination of LLLT and conventional therapy would result in a >50% reduction in licking visual analog score (LVAS) than conventional therapy alone. Dogs were randomly assigned to two groups. The treatment group received LLLT with blue and red light-emitting diodes (LEDs), while control group had sham therapy (laser off). Treatments were three times weekly for two weeks, then twice weekly for two weeks for a total of 10 visits. All dogs received systemic antibiotics and trazodone. Descriptive statistics were performed (mean, standard deviation). There was an overall decrease in LVAS in both control and treated dogs. Treatment dogs had a 20% greater decrease in LVAS compared to control dogs. iii The use of LLLT as a non-invasive treatment resulted in an additional decrease in licking behavior in dogs with ALD. LLLT should be considered as an adjunctive treatment in the management of dogs with ALD. iv Dedication To my parents: Thank for your love and unending support. Without you both I would not have the work ethic and drive to accomplish all that I have. To my husband Jayme: Through all these stressful years we have finally made it to the end, but I hope this is just the beginning. Thank you for everything, I would not have made it to this point and become the person I am today without you by my side. v Acknowledgements I would like to thank my advisor, Lynette Cole, for all her help and guidance throughout this entire residency. You have always pushed me to be my best and your strong mentorship is the reason I was able to accomplish my goals. This thesis, and our mutual love for Cavies, will always tie us together. Thank you to my committee members, Sandra Diaz, Wendy Lorch, Joshua Daniels and Paivi Rajala-Shultz. Your friendship and support will be forever appreciated. Thank you to my resident-mates Stephanie Abrams, Melanie Hnot and Holly Roberts. You all kept me levelheaded, smiling and laughing. Thank you to Deb Crosier for all the cute cat videos and cat noses that made the days go by easier. vi Vita May 2004………………………………………..B.S. Animal Science, Cornell University May 2013……………………………………….D.V.M., University of Minnesota July 2014 to present……………......…………...Graduate Teaching and Research Associate, The Ohio State University Fields of Study Major Field: Comparative and Veterinary Medicine Studies in Dermatology vii Table of Contents Abstract…………………………………………………………………………...……….ii Dedication………………………………………………………………………...…...…..v Acknowledgements………………………………………………………………..……...vi Vita…………………………………………………………………………………....…vii List of Tables………………………………………………..……………………...….…xi List of Figures………………………………………….………………………………..xiii CHAPTER 1 Introduction…………………………………………………..……….....….1 CHAPTER 2 Literature Review..........................................................................................4 2.1 Acral lick dermatitis………………..…………………………………………....…….4 2.1.1 Disease Pathophysiology………………….……………………..……...…..4 2.1.1.1 Primary Etiologies……………………………………………...…4 2.1.1.2 Differential Diagnoses…...……………………………………......6 2.1.1.3 Predisposing Factors……………….……………………………...8 2.1.1.4 Perpetuating Factors……………………...……….………….........8 2.1.2 Clinical Signs………………………………………..…………………....…9 2.1.3 Diagnostic Approach…………………..………………………………........9 2.2 Treatment………………………………………………………………………….....12 2.2.1 Antibiotics………………………………………………………………….12 2.2.2 Topicals…………..………………………………………………………...13 2.2.3 Treatment of Pruritic Disease……………………………...…...……….....13 2.2.3.1 Glucocorticoids……………...…………………………...14 2.2.3.2 Janus-kinase Inhibitors……………………………......….14 2.2.3.3 Monoclonal Antibody Therapy…………………….…….15 2.2.4 Anti-anxiety Medications.......................................………………………...15 2.2.4.1 Serotonin Antagonist and Reuptake Inhibitor….....……...15 2.2.4.2 Tricyclic Antidepressants...........…………………………16 2.2.4.3 Selective Serotonin reuptake Inhibitors (SSRIs)......…….17 2.2.4.4 Narcotic Antagonists.......………………………………...18 viii 2.2.5 Laser Therapy............................................................……………………...18 2.2.6 Radiation Therapy..........................................................……………….….19 2.2.7 Surgery..................................................................................……………...20 2.2.8 Electronic Stimulation......................………………………………………21 2.2.9 Orgotein........………………………………………………………………21 2.2.10 Acupuncture.............……………………………………………………...22 2.3 Prognosis.......................……………………………………………………………...22 2.4 Photostimulation.......................……………………………………………………...22 2.4.1 Low-Level Light Therapy (LLLT)………………………....…...…………22 2.4.1.1 Classification..........................…………………………………....22 2.4.1.2 Mechanism of Action………….….……....……………………...23 2.4.2 Uses of LLLT……….……….................……......…………………............24 2.4.2.1 Human Medicine……………………..………...........…………..24 2.4.2.2 Veterinary Medicine……………………………………………..27 2.4.3 Light Emitting Diodes.........................………….………………………….29 2.4.3.1 Definition………………............................……………………...29 2.4.3.2 Classification..................................................................................30 2.5 Photodynamic Therapy……………...................…………………………………….33 CHAPTER 3 In vitro bactericidal activity of blue light (465-nm) phototherapy on methicillin-susceptible and methicillin-resistant Staphylococcus pseudintermedius…………………………………………………………………..….…35 3.1 Abstract…………………………...………………………………………….35 3.2 Introduction………………….....….…………………………………………36 3.3 Materials and Methods……………………………....……………………….38 3.3.1 Bacterial Isolates and Culture…………………………….………..38 3.3.2 Blue Light Therapy……........……………………………………...39 3.3.3 Statistical Analysis………………….......………………………….40 3.4 Results.................…………………………………………………………….40 3.4 Discussion.......................................................................................................,41 CHAPTER 4 Low-level laser therapy as an adjunctive treatment for canine acral lick dermatitis: A randomized, double-blinded, sham-controlled study…………….......…....51 4.1 Abstract………………………………....…………………………………....51 4.2 Introduction…………………………………………......…………………....52 4.3 Materials and Methods…………………………………...…………………..54 4.3.1 Animals....................................………………………………….....55 4.3.2 Study Design...................…………………………………………..56 4.3.3 Laser Treatment……………………………………...........….…....59 4.4 Results.……………………………........................................……….……....60 4.4.1 Signalment and History.…………………………………….....…...60 4.4.2 Diagnostic Tests.…………………........................………………...61 4.4.2.1 Deep Skin Scrapings
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