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The Effects of X-Irradiation on Embryogenesis, Infectivity, and Migratory Behavior of the Larvae of Toxocara Canis (Werner, 1782) in White Mice

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The Effects of X-Irradiation on Embryogenesis, Infectivity, and Migratory Behavior of the Larvae of Toxocara Canis (Werner, 1782) in White Mice This dissertation has been microfilmed exactly as received 70-6825 LYLES, D.V.M., Demetrice Irving, 1922- THE EFFECTS OF X-IRRADIATION ON EMBRYOGENESIS, INFECTIVITY, AND MIGRATORY BEHAVIOR OF THE LARVAE OF TOXOCARA CANIS (WERNER, 1782) IN WHITE MICE. The Ohio State University, Ph.D., 1969 Veterinary Science University Microfilms, Inc., Ann Arbor, Michigan THE EFFECTS OF X-IRRADIATION ON EMBRYOGENESIS, INFECTIVITY, AND MIGRATORY BEHAVIOR OF THE LARVAE OF TOXOCARA CANIS (WERNER, 1782) IN WHITE MICE DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University Demetrice Irving Lyles, D.V.M., M.S. The Ohio S ta te U niversity 1969 Approved by Adviser Veterinary Parasitology ACKNOWLEDGMENTS This work is dedicated in memory of my late adviser, Professor Fleetwood R. Koutz, who more than anyone else, encouraged my work in Veterinary Parasitology and gave much of his valuable time advising me on this research. I would like to sincerely thank Professor Walter G. Venzke for assuming the position as my adviser in the absence of Dr. Koutz and also for inspiration concerning the endocrinology phase of the pre­ liminary research. In regard to the latter, thanks are extended to Professor Thomas E. Powers of the Department of Veterinary Physiology and Pharmacology for steroid dosage suggestions. Sincere gratitude is extended to Professor Willard C. Myser of the Department of Zoology and Entomology and to Assistant Professor James K. Burt of the Department of Veterinary Radiology for help with the radiological techniques used. Completion of this work could not have been accomplished without their assistance. Sincere thanks are extended to Assistant Professor Marion W. Scothom, Acting Chairman, of the Department of Veterinary Parasitology, for information gained from his "storehouse" of experience on certain phases of this research. To Professor Harold Groves, Department of Veterinary Parasitology, sincere thanks are extended for help in taking colored photographs. The time spent "just talking" to Professor Carl E. Venard regarding this research and related subjects was appreciated and stimulating. Acknowledgment is due the Department of Health, Education, and Welfare, National Institutes of Health for financial support for this resea rch . I would like to express my gratitude to my wife, Peggy, and to my children whose patience and encouragement throughout' the last several years contributed immeasurably to the successful completion of this research . iii VITA August 2, 1922 Bora - Sawyerville, Alabama 1940-1942 . Study toward B.Sc., Tuskegee Institute, Alabama 1942-1945 . U. S. Military Duty. World War II, Southwest Pacific Ar 1949 . D.V.M., Tuskegee Institute, Alabama 1949 . Small animal practitioner Louisville, Kentucky 1949-1956 . Instructor of Pathology and in charge of Necropsy Laboratory, School of Veterinary Medicine, Tuskegee I n s t i tu t e , Alabama 195S • . M.S., The Ohio State university, Columbus, Ohio 1958-1966 . Assistant Professor of Pathology and Parasitology and in charge of Clinical Diagnostic Laboratory, School of Veterinary Medicine, Tuskegee Institute, Alabama. PUBLICATIONS Eperythrozoonosis in a Herd of Purebred Landrace Pigs. AVMA 135 (1959): 226-228 Ecology of Filarial Nematodes. Tuskegee Veterinarian 3 No. 3 (1959) Eimeria acervulina and Eimeria maxima Infection in Chickens (Callus domesticus) Case Report. Tuskegee Veterinarian 6 No. 2 (19625 iv FIELDS OF STUDY Major Field: Veterinaiy Parasitology Studies in Advanced Veterinary Parasitology. Professors Fleetwood R. Koutz and Harold F. Groves Minor Problems in Veterinary Parasitology. Professors Fleetwood R. Koutz and Harold F. Groves Studies in Principles of Infection, Resistance, and Serology. P rofessor M. C. Dodd. Studies in Pathogenic Protozoology. Professor Julius P. Kreier Studies in Veterinary Endocrinology. Professor Walter G. Venzke Minor Problems in Veterinary Clinical Laboratory Diagnosis. P rofessor W alter F. Loeb Studies in Germfree and Gnotobiotic Animals. Associate Professor Donald B. Gisler Studies in Histological Techniques. Professor Martin Y. Andres Studies in Radiation Biophysics. Dr. William G. Myers Studies in Acarology. Institute of Acarology, University of Maryland. Professors G. W. Wharton, C. E. Younker, J. H. Camin, Edward W. Baker, and G. Anastas Studies in Medical Entomology. Professor Carl E. Venard. Minor Problems in Parasitology Technique. Professor J. N. Miller Studies in Laboratory Diagnosis of Rabies and Veterinary Mycology. U. S. Public Health Services, CDC, Atlanta, Georgia M.S. Thesis - Morphology and Periodicity of M icrofilaria in Dogs v TABLE OF CONTENTS Page ACKNOWLEDGMENTS............................. i i VITA ........... iv FIELDS OF STUDY ...................................................................................................... v LIST OF TABLES „ v i i i LIST OF ILLUSTRATIONS/. .................................. .... ..................... 3d INTRODUCTION......................... 1 LITERATURE REVIEW ............................................... .......................... 5 NoHi6ncXsijuil6 . ............o.......... 5 Classification ...................a... 5 Life cycle and development of Toxocara canis .......... 6 Infectivity .......................................................... 7 Life cycle and development of larvae in the normal host (dog) 8 Migratory behavior of Ascarid larvae in mice ................. 12 Invasion of central nervous system ........ ................. 15 Morphology of the second-stage larvae ........... 17 X-ray and irradiation effects ............... 17 1. Historical ................................ ...... 17 Effects of radiation on eggs of helminths 18 Effects of irradiation on larvae and adult helminths .... 27 MATERIALS AND METHODS ................................. 36 Collection of worms and harvesting of eggs for studying the effects of x-rays of embryogenesis ..... ...................... 36 Collection of worms and harvesting of eggs for infectivity av.^ migratory studies ....... ............ 37 Irradiation procedures for studying the effects of x-rays on embryogenesis.......................................................... 38 Irradiation procedures for studying the effects of x-rays on infectivity and migratory behavior of Toxocara canis larvae. 39 Experimental animals .......................................... 40 Preparation of egg suspension for mouse intubation ................. 40 Egg intubation procedures ........ 41 Necropsy and tissue digestion procedures . .......................... 42 vi TABLE OF CONTENTS (Continued) Page RESULTS OF EXPERIMENTS DEALING WITH THE EFFECTS OF X-RAYS ON EMBRYOGENESIS OF TOXOCARA CANIS...................................... 7 .................... 49 Experiment 1 Development of Toxocara canis eggs . 49 Development of rionirradiated Toxocara canis eggs ......... 49 Experiment 2 Development of eggs x-irradiated at 5>000 r. 51 Experiment 3 Development of Toxocara canis eggs x-irradiated at 10,000 r ........................................................... 51 Experiment 4 Development of Toxocara canis eggs x -irra d ia te d a t 20,000 r . .................................................. 53 Experiment 5 Development of Toxocara canis eggs x-irradiated at 40,000 r . .................................................. 56 Experiment 6 Development of Toxocara canis eggs x-irradiated at 80,000 r .................................................................... 56 RESULTS OF EXPERIMENTS DEALING WITH THE EFFECTS OF X-RAYS ON TOXOCARA CANIS LARVEL MIGRATION IN MICE .................................. 77 Experiment 1..... ....................................................................... 77 Experiment 2 ............................................................................................ 81 Experiment 3 ............................................................................................ 83 Experiment 4 ....................................................................................... • 65 Experiment 5 ............................................................................................ 67 Experiment 6 ............ .......................................... 88 Trend S ta stic ........................................................................................ 113 Null hypothesis .................................................................................... 113 Alternative hypothesis .................. 113 RESULTS OF PRELIMINARY EXPERIMENTS ...................................... 125 DISCUSSION......................................... ............................................................... 125 Embryogenesis.............................................................. 125 Migratory Behavior . ....................................................................... 131 SUMMARY................................................................................................................. 139 LITERATURE C IT E D ................................................................................... 142 vii LIST OF TABLES Table Page 1 The design of experiment to study the effects of x-irradiation on embryogenesis of the eggs of Toxocara canis 43 2 The design of experiment using nonirradiated and x-irradiated embryonated eggs of Toxocara canis ....................... 44 3 The development of nonirradiated eggs of Toxocara canis at room temperature (29° - 30° C.) (Developmental stages in p e r c e n t ) ..............................................................................................................50
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