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Some Immunological and Other Studies in Mice on Infection with Embryonated Eggs of Toxocara Canis (Werner, 1782)

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Some Immunological and Other Studies in Mice on Infection with Embryonated Eggs of Toxocara Canis (Werner, 1782) This dissertation has been 69-11,668 microfilmed exactly as received MALIK, Prem Dutt, 1918- SOME IMMUNOLOGICAL AND OTHER STUDIES IN MICE ON INFECTION WITH EMBRYONATED EGGS OF TOXOCARA CANIS (WERNER, 1782). The Ohio State University, Ph.D., 1968 Agriculture, animal pathology Health Sciences, immunology University Microfilms, Inc., Ann Arbor, Michigan SOME IMMUNOLOGICAL AND OTHER STUDIES IN MICE ON INFECTION WITH EMBRYONATED EGGS OF TOXOCARA CANIS (WERNER, 1782) DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Prem Dutt Malik, L.V.P., B.V.Sc., M.Sc ****** The Ohio State University 1968 Approved by Adviser / Department of Veterinary Parasitology ACKNOWLEDGMENTS I wish to express my earnest thanks to my adviser, Dr. Fleetwood R. Koutz, Professor and Chairman, Department of Veterinary Parasitology, for planning a useful program of studies for me, and ably guiding my research project to a successful conclusion. His wide and varied experience in the field of Veterinary Parasitology came handy to me at all times during the conduct of this study. My grateful thanks are expressed to Dr. Harold F. Groves, for his sustained interest in the progress of this work, and careful scrutiny of the manuscript. Thanks are extended to Dr. Walter G. Venzke, for making improvements in the manuscript. Dr. Marion W. Scothorn deserves my thanks for his wholehearted cooperation. To Dr. Walter F. Loeb, I am really indebted for his valuable time in taking pictures of the eggs, the larvae, and the spermatozoa of Toxocara canis. The help of Mr. William A. Bruce, a graduate student in acarology, in identifying and taking pictures of Myocoptes musculinus mites, is acknowledged. My thanks also go to my friend and fellow student, Dr. Demetrice I. Lyles, for his valuable discussions and mutual help. Lastly, I bow my head in solemn prayer for those cute, and lovely little mice whom I layed to eternal rest to satisfy man's hunger for the unknown. VITA August 15, 1918 Born - Multan, India (now Pakistan) 1941 .......... L.V.P., Punjab Veterinary College, Lahore 1942-1957. Veterinary Assistant Surgeon, Civil Veterinary Department, Punjab, India. 1952 .......... B.V.Sc., Punjab College of Veterinary Science and Animal Husbandry, Hissar, India. 1955 ........... Advanced training in rabies research, Central Research Institute, Kasuli, India. 1956 , ........ Post-graduate course in Veterinary Science, Indian Veterinary Research Institute, Mukteswar/Izatnagar, U.P., India. 1958-1960. Research Assistant, College of Veterinary and Animal Science, Hissar, India. 1960-1965. Assistant Disease Investigation Officer (Camels) Punjab, College of Veterinary and Animal Science, Hissar, India. 1966 .......... M.Sc., The Ohio State University, Columbus, Ohio. PUBLICATIONS "Amphistomiasis in Cattle in the Punjab." Journal of Animal Health, Calcutta, 1960. "Incidence, Diagnosis, and Control of Immature Amphistomiasis in the Punjab." Conference on Parasites and Parasitic Diseases, Indian Council of Agricultural Research, 1961. "Parasitic Gastro-enteritis in India with Particular Reference to the Main Pathogenic forms and Their Control." 11th All India Confer­ ence on Parasitic Diseases, Indian Council of Agricultural Research, 1964. "Salmonella Serotypes from Camel in India." Journal of Research, P.A.U., Ludhiana, 1967. iii VITA (continued) Four Annual Technical Reports on "Normal Worm Burden, and Normal Fecal Egg Output Rate in Sheep, Goats, Cattle, and Buffaloes in the Punjab." Submitted to the Indian Council of Agricultural Research, New Delhi, 1957-1960. Five Annual Technical Reports on "The Diseases of Camels in the Punjab. Submitted to the Indian Council of Agricultural Research, New Delhi 1960-1965. FIELDS OF STUDY Major Field: Veterinary 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 and Resistance. Professor M. C. Dodd Studies in Serology. Professor F. W. Chorpenning Studies in Pathogenic Protozoology. Professor Julius P. Kreier Studies in Veterinary Endocrinology. Professor Walter G. Venzke Studies in Medical Entomology. Professor Carl E. Venard Studies in the Prevention and Control of Diseases of Poultry. Professor Clyde A. Marsh Minor Problem in Veterinary Clinical Laboratory Diagnosis. Professor Walter F. Loeb Minor Problem in Zoology (Molluscs). Professor David H. Stansberry. TABLE OF CONTENTS Page ACKNOWLEDGMENTS ................................................... ii VITA ................................. iii LIST OF T A B L E S ................................................... vii LIST OF ILLUSTRATIONS ............................................viii INTRODUCTION ..................................................... 1 Objectives............................................ 3 LITERATURE REVIEW ............................................... 5 Nomenclature ............................................... 5 Classification ............................................ 3 Historical review ........................................ 6 Life h i s t o r y ............................................... 8 Prenatal infection ........................................ 9 Migratory behavior ........................................ 9 Survival of larvae in host t i s s u e s .......................... 11 Central nervous system involvement ........................ 11 Morphology of second-stage larvae ....................... 14 MATERIALS AND METH O D S ........................................ 16 Collection of e g g s .......................................... 16 Collection of eggs from fe c e s .............................. 17 Collection of parasites from hosts ....................... 17 Collection of eggs from parasites .......................... 18 1. Maceration of the whole w o r m s ..................... 18 2. Dissection of the u t e r i ............................18 3. (a) Natural oviposition............................20 (b) At incubator t e m p e r a t u r e ......................21 (c) Maintaining female worms with male worms . 21 (d) Maintaining worms in 1 per cent dextrose s o l u t i o n .......................................21 Experimental animals ...................................... 22 Preparation of inoculum ............................. 32 Experimental procedure ......................................33 Plan of s t u d y ............................................... 41 v TABLE OF CONTENTS (Continued) Page RESULTS ..................................................... 46 Longevity of larvae in tissues ........................ 47 Lethal effects of Toxocara canis eggs ................ 56 Lethal doses of Toxocara canis eggs .................. 60 Challenge experiments ................................. 65 Conclusions of challenge experiments .................. 70 Infectivity of 27-month-old Toxocara canis eggs .... 71 Lethal effects of 27-month-old Toxocara canis eggs . 73 Challenge experiments with 27-month-old Toxocara canis e g g s ................................................. 73 Treatment and control of murine mange .......... 76 Toxocara canis spermatozoa . ........................ 77 DISCUSSION................................................... 80 Criteria of immunity inhelminth i n f e c t i o n s ........... 80 Lethal doses ........................................... 82 Resistance to reinfection .......................... 83 Prepatent period ....................................... 88 The effect of storage on viability of Toxocara canis e g g s ................................................. 90 SUMMARY ................................................... 91 LITERATURE CITED ............................................. 95 vi LIST OF TABLES Table Page 1 The design of experiment to determine the lethal effect of different doses of 30-day-old embryonated eggs of Toxocara canis by oral intubation............................. 42 2 The design of experiment for determining the minimum lethal dose (M.L.D.) and lethal dose 50 (L.D. 50) of Toxocara canis 30-day-old embryonated eggs for mice by oral intubation . 43 3 The design of experiment to determine the minimum immuniz­ ing dose (M.I.D.) of Toxocara canis eggs for m i c e ...........44 4 The design of experiment to test the infectivity of Toxocara canis eggs stored in a refrigerator for 27 m o n t h s ........... 44 5 The design of experiment to determine the lethal effects of ■27-month-old Toxocara canis eggs for m i c e ................... 45 6 Lethal effects of 30-day-old embryonated eggs of Toxocara canis for mice by oral i n t u b a t i o n ........................... 57 7 Minimum lethal dose of 30-day-old embryonated eggs of Toxocara canis for mice by oral intubation................... 63 8 Lethal dose 50 (L.D. 50) of Toxocara canis eggs for mice by oral intubation..................................... 64 9 Results of challenge of surviving mice with 40,000 eggs of Toxocara canis four weeks after primary infection ........ 68 10 Results of challenging previously infected mice with 15,000 and 20,000 eggs of Toxocara canis three weeks after the primary infection ........................... 69 11 Distribution of Toxocara canis larvae in white mice tissues infected orally with 27-month-old eggs stored in a refrigerator ............................................... 72 12 Lethal effects of 27-month-old embryonated eggs of
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