INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. in the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6” x 9" black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. UMI Bell & Howell Information and Learning 300 North Zeeb Road, Ann Aitor, Mi 48106-1346 USA 800-521-0600 IMMUNE RESISTANCE TO RHIPICEPHALUS SANGUINEUS IN DOGS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Sathaporn Jittapalapong, D.V.M. The Ohio State University 1999 Dissertation Committee: Approved by Professor John C. Gordon, Adviser Adviser Professor Roger W. Stich, Co-Adviser Professor Lawrence E. Mathes Co-Adviser Professor Daral J. Jackwood Veterinary Preventive Medicine Program UMI Number: 9941352 Copyright 1999 by Jittapalapong, Sathaporn AH rights reserved. UMI Microform 9941352 Copyright 1999, by UMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arbor, MI 48103 ABSTRACT Rhipicephalus sanguineus is a cosmopolitan tick parasite of dogs that also transmits several protozoal and rickettsial pathogens. Host Immunity to ticks would be an alternative to acarlcldes for the control of ticks and tickborne diseases, but the ability of dogs to acquire resistance against R. sanguineus has been questioned. The purpose of this study was to ascertain the effects of Immunity Induced In dogs with tick salivary gland (TSG) or mIdgut (TMG) antigens compared to naturally acquired host resistance Induced by repeated Infestations. Parameters of tick feeding and fecundity were then determined for subsequently challenge fed female ticks. Female ticks fed on dogs Immunized with TSG had the lowest engorgement weight of all three groups. A delayed feeding period and a decreased number of engorged females Indicated resistance In the TSG-lmmunlzed group. Reduced fecundity was Indicated In ticks fed on dogs Immunized with TMG by prolonged periods of pre-ovlposltlon, oviposltlon, and egg Incubation In addition to reduced egg production and viability, compared to that of ticks fed on naïve or TSG- lmmunlzed dogs. ELISA results Indicated cross-reactlvlty between sera collected from dogs Immunized with either TSG or TMG for either antigen preparation. Western blot analysis demonstrated nine bands of salivary gland antigens, ranging from 10 kDa to 100 kDa, which were recognized by either anti-TSG or TMG antibodies. Seven protein bands, at 450, 250, 210, 130, 125, 105, and 38 kDa, were uniquely recognized by sera from the TSG-immunized dogs. Immunoblotting results indicated that antibodies specific for TSG might be associated with lowered female body weight at engorgement, an important feeding performance parameter that directly effects tick fecundity. At least nine protein bands, ranging from 13 to 210 kDa, found in midgut, reproductive organ and egg antigens were uniquely recognized by sera from TMG- immunized dogs that suppressed tick fecundity. This work represents the first essential step to characterize tick antigens that are associated with artificial O induction of reduced tick performance. Immunization of purified antigens associated with reduced tick feeding and fecundity performances may lead to greater resistance against R. sanguineus infestation in dogs. I l l Dedicated to PHO and SOMPORN JITTAPALAPONG IV ACKNOWLEDGMENTS I would like to express most of my gratitude and appreciation to two persons, Dr. Omar O. Barriga (a former academic advisor), who give me inspiration and intellectual support and Dr. Roger W. Stich for encouragement and enthusiasm which made this dissertation possible and helped support me in the last portion of this work. 1 also thank Dr. John C. Gordon, Dr. Cliff Monahan, Dr. Lawrence E. Matthes, Dr. Thomas E. Wittum, Dr. Daral J. Jackwood and Dr. Glen Needham for their valuable suggestions. I am grateful to Catherine A. Bremer for the excellent technical assistance as well as staff and faculties of Veterinary Preventive Medicine for their helps and supports. This research was funded by the Morris Animal Foundation, the Graduate School Alumni Research Award (The Ohio State University) and Kasetsart University Research and Development Institute of Thailand. Lastly, I wish to thank. Royal Thai Government to support for the scholarship, and Dr. Weeraphol Jansawan, the chief of Parasitology Department, Kasetsart University for their concern with my studies. I would like to dedicate this work to my father, mother and family with my thanks for their support throughout my education. VITA February 14, 1959. Born-Nonthaburi, Thailand 1981....................... B.Sc. Kasetsart University, Thailand 1983....................... D.V.M. Kasetsart University, Thailand 1983-1988............. Instructor, Faculty of Veterinary Medicine, Kasetsart University, Thailand 1988-1993............. Assistant Professor, Faculty of Veterinary Medicine, Kasetsart University, Thailand 1993-Present ......... Graduate student. The Ohio State University 1998-1999............. Graduate Teaching and Research Assistant, Department of Veterinary Preventive Medicine, The Ohio State University VI PUBLICATIONS 1. Jittapalapong, S.1981. Coumarin poisoning in Dogs. Journal of Thai Veterinary Practitioner 3 (4): 263-273. 2. Jittapalapong, S.,Jansawan, W., Sriworanat, P., and Daengsupha, P. 1986. Infection Rate of Ascaris iumbricoides in pigs. Journal of Kasetsart University Veterinary Medicine 7(3): 144-153. 3. Chantharayotha, 0., Khuphophan, S., Jansawan, W., Sriworanat, P., and Jittapalapong, S., 1986.Intestinal Parasites Causing Piglet Diarrhea in Four Provinces in Thailand. The Annual Report of Kasetsart University. 121. 4. Jittapalapong, S.,Jansawan, W., and Pinyopummin, T. 1987. Epidemiological Study of Dairy Cow in Nongpho. Journal of Kasetsart University Veterinary Medicine 8 (1): 21-29. 5. Jittapalapong, S.,Jansawan, W., and Pinyopummin, T. 1987. Epidemiological Study of Calves in Nongpho. Journal of Kasetsart University Veterinary Medicine 8 (1): 124-132. 6. Sunthrasamai, P., Warrell, M. J., Warrell, D. A., Chanthavanich, P., Looareesuwan, S., Supaphchana, A., Phanuphak, P., Jittapalapong, S., Yager, P. A., and Baer, G. M. 1987. Early Antibody Responses to Rabies Post-exposure Vaccine Regimens. American Joumai of Tropical Medicine and Hygiene 36{^): 160-165. 7. Jittapalapong, S.,Jansawan, W., Sriworanat, P., and Daengsupha, P. 1990. Survey Natural and Experimental Reservoir Host of Capiiiaria phiiippinensis in Pet and Wild Animals. Joumai of KU Veterinary Hospital 3(1): 69-77. 8. Jittapalapong, S.1990. Epidemiological Study of HeartWorm Disease in Dogs in Bangkok and Vicinity. Joumai of Thai Veterinary Practitioner^2{4): 265-277. 9. Jittapalapong, S.,and Tipsawek, S. 1991. Epidemiological Survey of Blood Protozoa and Rickettsia in Dogs in Bangkok and Vicinity. Kasetsart University Joumai25{^): 75-82. 10. Jittapalapong, S.,and Jansawan, W. 1993. Preliminary Study of Blood Parasites in Cats in Bangkok. Kasetsart University Journal 27(3): 330-335. VII 11. Jansawan, W., Jittapalapong, S.,and Jantharat, N. 1993. Effect of Stemona collinsae Extract against Cattle Ticks {Boophilus microplus). Kasetsart University Joumai 27(3): 336-340. 12. Jittapalapong, S.,Intharaksa, Y., Jantharat, N., and Phatthanathanang, K. 1993. Epidemiological Survey of Intestinal Parasites of Calves in the Northeastern Provinces of Thailand. Kasetsart University Journal 27(4): 469-473. 13. Jittapalapong, S.,Stich, R. W., Gordon, J. C., Wittum, T. E., and Barriga, O. O. 1999. Reduced Feeding and Fecundity Performance of Rhipicephalus sanguineus (Acari: Ixodidae) by Salivary Gland or Midgut Immunizations, and Repeated Infestations in Dogs (Submitted) 14. Jittapalapong, S.,Stich, R. W., Gordon, J. C., Bremer C. A, and Barriga, O. O. 1999. Humoral Immune Response in Dogs previously Immunized with Salivary gland. Midgut or Repeated Infestations with Rhipicephalus sanguineus. (New York Academy of Science:In press) FIELDS OF STUDY Major field: Veterinary Preventive Medicine VIU TABLE OF CONTENTS Pages Abstract......................................................................................................... li Dedication..................................................................................................... iv