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Endogenous Development of Two Coccidian Parasites : Isospora Arctopitheci and Sarcocystis Hemionilatrantis

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Endogenous Development of Two Coccidian Parasites : Isospora Arctopitheci and Sarcocystis Hemionilatrantis University of Montana ScholarWorks at University of Montana Graduate Student Theses, Dissertations, & Professional Papers Graduate School 1982 Endogenous development of two Coccidian parasites : Isospora arctopitheci and Sarcocystis hemionilatrantis Andrew T. Olcott The University of Montana Follow this and additional works at: https://scholarworks.umt.edu/etd Let us know how access to this document benefits ou.y Recommended Citation Olcott, Andrew T., "Endogenous development of two Coccidian parasites : Isospora arctopitheci and Sarcocystis hemionilatrantis" (1982). Graduate Student Theses, Dissertations, & Professional Papers. 7328. https://scholarworks.umt.edu/etd/7328 This Thesis is brought to you for free and open access by the Graduate School at ScholarWorks at University of Montana. It has been accepted for inclusion in Graduate Student Theses, Dissertations, & Professional Papers by an authorized administrator of ScholarWorks at University of Montana. For more information, please contact [email protected]. COPYRIGHT ACT OF 1976 This is an unpublished manuscript in which copyright sub­ sists. Any further reprinting of its contents must be approved BY the author. MANSFIELD L i b r a r y Un i v e r s i t y of Mo n t a n a Date : -■ 1 8 8 3 ____ ENDOGENOUS DEVELOPMENT OF TWO COCCIDIAN PARASITES ISOSPORA ARCTOPITHECI AND SARCOCYSTIS HEMIONILATRANTIS by Andrew T. Olcott B.A., University of Montana, 1976 Presented in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE UNIVERSITY OF MONTANA 1982 Chairman, £oard of Examiners rean. Graduate School Date UMI Number: EP38129 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note wilt indicate the deletion. UMT DtssMtation PüWwkhing UMI EP38129 Published by ProQuest LLC (2013). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code ProQuesf ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106 - 1346 Olcott, Andrew T., M.S. March, 1982 Microbiology Endogenous Stage of Two Coccidian Parasites: Isospora arctopitheci and Sarcocystis hemionilatrantis Director: C. A. Speer Û 4S Development of Isospora arctopitheci and Sarcocystis hemionilatrantis was studied by light, and scanning (SEM) and transmission (TEM) electron microscopy. Thirteen marmosets (Saguinus geoffroyi) were experimentally inoculated with sporulated oocysts of Isospora arctopitheci. Parasites were found at 1—7 days post inoculation (DPI) in epithelial cells of the villi of the small intestine, primarily the jejunum. No parasites were found in any extraintestinal tissue examined. Sporozoites invaded host cells and divided several times by endodyogeny to form as many as 16 zoites (3.7 x 1.5 pm) within a single parasitophorous vacuole. Usually, zoites es­ caped from host cells and entered other cells to undergo gametogony. Occasionally, zoites remained in the same parasitophorous vacuole and underwent gametogony. Microgamonts were 12.1 ym in diameter and con­ tained about 50—100 microgametes. Macrogamonts were 11.7 x 14.9 ym. An eosinophilic body (EB) was associated with the surface of micro- and macrogamonts. In mature microgamonts, the EB usually remained with the centrally located residual body, whereas in zygotes it appeared to be incorporated into the oocyst wall. Oocysts were present at 7 DPI, meas­ ured 14.2 X 18.1 ym, and were passed unsporulated. Sloughing of the intestinal epithelium occured at 7 DPI. Four of the 13 marmosets died during the infection; one at 3 and 5 DPI, and 2 at 7 DPI. Zygotes and oocysts of Sarcocystis hemionilatrantis were studied in the lamina propria of the small intestine of coyotes (Canis latrans) by SEM and TEM. Zygotes and oocysts did not occur in a parasitophorous vacuole but were only partially enveloped by host cell pseudopodia. Zygotes were spheroid, and had a thin oocyst wall. Wall forming bodies typical of Coccidia were not seen. Lipid bodies exocytosed into a space between the oocyst wall and the sporont pellicle, much of which adhered to the inner surface of the oocyst wall. Membrane layers and zones of granular material coalesced to form separate inner and outer layers of the sporont. These layers invaginated to separate the sporont into 2 sporoblasts. Mature oocysts contained 2 sporocysts, each with 4 sporo­ zoites. 11 ACKNOWLEDGMENTS Throughout the planning, investigation, and evaluation of this research and the preparation of this manuscript. Dr. C.A. Speer has been of the utmost help and professional guidance. I greatly appreciate his assistance and his serving as my thesis advisor. I also want to thank the members of my degree committee. Dr. J. Nakamura, Dr. R.A. Faust, and Dr. J. Ball, for their evaluation of this manuscript. I l l TABLE OF CONTENTS Page ABSTRACT.......................................................... ii ACKNOWLEDGMENTS.................................................. iii LIST OF PLATES................................................... vi ABBREVIATIONS.................................................... ix INTRODUCTION..................................................... 1 Taxonomie Classification of Isospora arctopitheci and Sarcocystis hemionilatrantis............................... 1 Besnoitia................................................... 2 Eimeria..................................................... 3 Frenkel ia................................................... 5 Hammond ia................................................... 6 Isospora.................................................... 7 Sarcocystis................................................. 9 Toxoplasma.................................................. 13 ENDOGENOUS DEVELOPMENT OF ISOSPORA ARCTOPITHECI................ 15 Materials and Methods....................................... 15 Results..................................................... 17 Location and Incidence.................................. 17 Asexual development...................................... 17 Sexual development....................................... 18 Oocysts.................................................. 20 Pathology................................................ 20 iv Page Plates...................................................... 22 Discussion.................................................. 27 ENDOGENOUS DEVELOPMENT OF SARCOCYSTIS HEMIONILATRANTIS........ 30 Materials and Methods...................................... 30 Results..................................................... 32 Transmission Electron Microscopy........................ 32 Scanning Electron Microscopy............................ 33 Plates...................................................... 37 Discussion.................................................. 46 LITERATURE CITED................................................. 51 LIST OF PLATES Plate Page 1. Endogenous stages of Isospora arctopitheci............. 22 1. Sporozoite 2. Zoites 3. Zoites 4. Zoites 5. Zoites 6. Zoites 7. Macrogamont 8. Macrogamont 9. Macrogamont 10. Zygote 11. Macrogamont 12. Macrogamont and Zoites 2. Endogenous stages of Isospora arctopitheci............. 24 13. Micro gamont 14. Zoite and microgamont 15. Microgamont 16. Microgamont and microgamete 17. Oocyst 18. Macrogamont 19. Macrogamont and microgamont 20. Pathology VI Plate Page 3. Scanning electron microscopy of the sexual stages....... 26 21. Microgamont and microgametes 22. Macrogamont 23. Eosinophilic body 24. Eosinophilic body 4. Oocysts of Sarcocystis hemionilatrantis.................. 37 25. Oocyst 26. Lipid body 27. Enveloping pseudopodia 5. Formation of wall around zygote and sporoblasts......... 39 28. Granular zones 29. Amylopectin and lipid bodies 30. Lipid extrusion and oocyst wall 31. Membrane fragments 6. Scanning electron microscopy of zygotes and oocysts 41 32. Zygote 33. Zygote 34. Oocyst 35. Oocyst 36. Oocyst containing 2 sporoblasts 37. Porous inner sporocyst wall 7. Sporulation of oocyst..................................... 43 38. Inner and outer sporocyst walls 39. Sporocysts 40. Cleavage of oocyst vii Plate Page 8. Single sporocyst in cross section....................... 45 41. Sporocyst viix ABBREVIATIONS In Text DPI days post inoculation DAI days after inoculation EB eosinophilic body M1-M6 membrane layers NBR National Bison Range OW oocyst wall SEM scanning electron microscopy TEM transmission electron microscopy v/v volume to volume WFB wall forming body w/v weight to volume “ 6 pm micrometer, 10 meter In Figures Am amylopectin B1 basal lamina Bv blood vessel Cc concavity Cg collagen Db dense body Eb eosinophilic body En endothelial cell ix FI flagellum Fx finger-like extension of Eb G1 granular layer Go Golgi complex Hn host cell nucleus Lb lipid body LI electron lucent layer between oocyst and sporocyst walls Lu lumen of intestinal tract Ma macrogamont Mb membrane fragment Mi microgamont in Isospora chapter Mi mitochondrion in Sarcocystis chapter No nucleolus Np nuclear pore Nu nucleus Oo oocyst Ow oocyst wall Pd pseudopodium Pv parasitophorous vacuole Rb residual body Rr rough endoplasmic reticulum Sp sporozoite Sr smooth endoplasmic reticulum Swl outer sporocyst wall Sw2 inner sporocyst wall Zo zoite xi INTRODUCTION Taxonomic Classification of
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