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ISOLATION, ANATOMICAL DISTRIBUTION and ANTIFUNGAL SUSCEPTIBILITY of DERMATOPHYTES AFFECTING HORSES in KWARA STATE, NIGERIA by RA

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ISOLATION, ANATOMICAL DISTRIBUTION and ANTIFUNGAL SUSCEPTIBILITY of DERMATOPHYTES AFFECTING HORSES in KWARA STATE, NIGERIA by RA ISOLATION, ANATOMICAL DISTRIBUTION AND ANTIFUNGAL SUSCEPTIBILITY OF DERMATOPHYTES AFFECTING HORSES IN KWARA STATE, NIGERIA By RASHIDAT BOLANLE BALOGUN DEPARTMENT OF VETERINARY MICROBIOLOGY FACULTY OF VETERINARY MEDICINE AHMADU BELLO UNIVERSITY, ZARIA NIGERIA JUNE, 2015 i ISOLATION, ANATOMICAL DISTRIBUTION AND ANTIFUNGAL SUSCEPTIBILITY OF DERMATOPHYTES AFFECTING HORSES IN KWARA STATE, NIGERIA By RashidatBolanle BALOGUN, DVM 2010 (ABU) MSc/Vet Med/5162/2011-2012 A THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTER OF SCIENCE IN VETERINARY MICROBIOLOGY DEPARTMENT OF VETERINARY MICROBIOLOGY AHMADU BELLO UNIVERSITY, ZARIA NIGERIA JUNE, 2015 ii DECLARATION I declare that the work in this Thesis entitled “Isolation, anatomical distribution and antifungal susceptibility of dermatophytes affecting horses in Kwara State, Nigeria” has been carried out by me in the Department of Veterinary Microbiology under the supervision of Dr. (Mrs.) C. N. Kwanashie and Prof. H. M. Kazeem. The information derived from the literature has been duly acknowledged in the text and a list of references provided. No part of this thesis was previously presented for another degree or diploma at this or any other institution. RashidatBolanle BALOGUN ____________ _____________ Student Signature Date iii CERTIFICATION This Thesis entitled “ISOLATION, ANATOMICAL DISTRIBUTION AND ANTIFUNGAL SUSCEPTIBILITY OF DERMATOPHYTES AFFECTING HORSES IN KWARA STATE, NIGERIA” by RashidatBolanle BALOGUN meets the regulations governing the award of the degree of Master of Science of the Ahmadu Bello University, Zaria and is approved for its contribution to knowledge and literary presentation. Dr. (Mrs.) C. N. Kwanashie Chairman, Supervisory Committee Signature Date Prof. H. M. Kazeem Member, Supervisory Committee Signature Date Dr. (Mrs.) C. N. Kwanashie Head of Department Signature Date Prof. A. Z. Hassan Dean, School of Postgraduate Studies Signature Date iv DEDICATION This work is dedicated to, my husband and children. v ACKNOWLEDGMENTS I wish to express my sincere gratitude to the Almighty Allah (Subhanahuwata‟alah) for giving me the strength and vigor and to members of my supervisory committee, Dr. (Mrs.) C. N. Kwanashie and Prof. H. M. Kazeem for their guidance and encouragement throughout the course of this study. I am grateful to the staff members of the Department of Veterinary Microbiology, for their support and encouragement throughout the period of this study. My special thanks to Dr Paul Habila, DrAnkeli, Mallam Dodo, mallamTanko, mallamSalisu and HajiaSalamat for the help and concern they demonstrated to see the completion of this work. My profound gratitude is to my professional colleagues in the University of Ilorin, especially Dr G. Atoyebi, Dr A. Obalowu, Dr Adam Mohd., Dr L. Jegede, Dr Paul and my brother, Kabir who all assisted me during my numerous trips to seek permission to collect skin samples from horses in Kwara state. I am grateful for the understanding and support of all members of my family, my dad, Shaid, Fati, Aisha, Meena and espeacially my mom and sister, Shaidat who have been patient enough to take care of my children in my absence. My special thanks also go to Prof. Sackey, A.K.B., Dr. AdamuJibril, Dr. Alam L. and Dr. M.T. Salawudeen for their help and contributions toward the success of this work. I would also like to acknowledge the support and encouragement given to me by the Dean of Faculty of Veterinary Medicine, Unilorin, Prof. S. F, Ambali and Director of V. T. H. University of Ilorin, Prof. E. O. Oyedipe, I am very grateful. Finally, I give my most special word of deep appreciation to my loving husband in the person of Suleiman ZakariyauJimoh, who stood by me in all ramifications, I am deeply indebted, and to my children (Faiza, Suleiman and Mohd- Khalid) for their patience and understanding. vi ABSTRACT Dermatophytes are fungi that colonize keratinized tissues of humans and animals. This study was conducted to isolate dermatophytes from both clinical and asymptomatic cases of dermatophytosis in horses in seven Local Government Areas (LGA) of Kwara State and to determine the susceptibility pattern of the dermatophytes isolated from different anatomical sites on horses to five antifungal agents. Ninety-one samples of plucked hair or scales and scrapings from the skin of horses were initially examined directly using microscopy prior to culture for isolation and identification usingSabouraud‟s dextrose agar (SDA), with 5% NaCl and potato dextrose agar as media for dermatophyte culture and isolation, respectively. Identification of each isolate was through observation of colonial morphology and microscopic appearance of lactophenol cotton blue stained fungal specimen obtained from culture. The assessment of antifungal susceptibility patterns of the dermatophytes isolated was by broth microdilution assay using ketoconazole, fluconazole, amphotericin B, griseofulvin and terbinafine as antifungal agents. From all the samples obtained (91 samples), 14(15.4%) were dermatophytes out of which one was from an asymptomatic horse. These dermatophytes were identified as members of Trichophyton(T.)andMicrosporum(M.)genera. The dermatophyte species isolated were T. tonsurans(7.14%) and T. soudanense (7.14%) which are anthropophylicdermatophytes, T. verrucosum(35.7%), M. gypseum (7.14%), M. persicolor (14.2%), M. equinum (7.14%) and M. fulvum (21.4%). Twelve of the 14 dermatophytes were isolated from 85 male horses while the remaining two were from six female horses. Based on anatomical sites, the highest isolation rate was from the limbs (18.7%) and the lowest from the abdomen (10%). However, the differences between the dermatophytes isolated from male and female horses or the different anatomical sites were not statistically significant (p ˃ 0.05). With regards to samples from the seven LGAs, vii samples collected from Ilorin-East LGAyielded the highest isolation rate (25%) whilst those fromBarutenLGA had the lowest isolation rate (9.1%). The antifungal susceptibility test showed that terbinafine was the most potent drug with the lowest range of MIC values (0.2-6.5µg/ml) followed by amphotericin B which had MIC range of 0.6-4.0µg/ml and then ketoconazole (0.3- 9µg/ml), whereas griseofulvin and fluconazole showed the highest MIC ranges of 1.5-8.0µg/ml and 0.6-19.2µg/ml, respectively,indicating that terbinafine was the most efficacious of the five antifungal agents used in this study. Culture and sensitivity tests should be carried out for effective diagnosis and treatment of fungal infections especially in horses. viii TABLE OF CONTENT Page Cover page -----------------------------------------------------------------------------------------------------i Title Page ------------------------------------------------------------------------------------------------------ii Declaration ---------------------------------------------------------------------------------------------------iii Certification --------------------------------------------------------------------------------------------------iv Dedication -----------------------------------------------------------------------------------------------------v Acknowledgements -----------------------------------------------------------------------------------------vi Abstract ------------------------------------------------------------------------------------------------------vii Table of Contents --------------------------------------------------------------------------------------------ix List of Tables ------------------------------------------------------------------------------------------------xii List of Figures -----------------------------------------------------------------------------------------------xiii List of Plates ------------------------------------------------------------------------------------------------xiv List of Abbreviations and Symbols -----------------------------------------------------------------------xv CHAPTER 1: INTRODUCTION------------------------------------------------------------------------1 1.1 Background of the Research ---------------------------------------------------------------------1 1.2 Statement of the Research Problem ------------------------------------------------------------4 1.3 Justification of the Study--------------------------------------------------------------------------5 1.4 Aim of the Study------------------------------------------------------------------------------------6 1.5 Objectives of the Study ---------------------------------------------------------------------------6 1.6 Research Questions --------------------------------------------------------------------------------7 CHAPTER 2: LITERATURE REVIEW---------------------------------------------------------------8 2.1 Fungal Infections----------------------------------------------------------------------------------8 2.2 Historical Background --------------------------------------------------------------------------10 2.3 Nomenclature-------------------------------------------------------------------------------------12 2.4 Virulence of Dermatophytes-------------------------------------------------------------------13 2.5 IsolationTechniques----------------------------------------------------------------------------14 2.6 Cultural Characteristics------------------------------------------------------------------------15 2.6.1 Identifying dermatophytes -----------------------------------------------------------------------15
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