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The Pathophysiology of Astrovirus Infection in Hatchling Turkeys

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The Pathophysiology of Astrovirus Infection in Hatchling Turkeys Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1992 The ap thophysiology of astrovirus infection in hatchling turkeys Mari Lynn Thouvenelle Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Veterinary Pathology and Pathobiology Commons Recommended Citation Thouvenelle, Mari Lynn, "The ap thophysiology of astrovirus infection in hatchling turkeys " (1992). Retrospective Theses and Dissertations. 9956. https://lib.dr.iastate.edu/rtd/9956 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly firom 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 corner 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. University Microfilms International A Bell & Howell Information Company 300 North Zeeb Road, Ann Arbor, Ml 48106-1346 USA 313/761-4700 800/521-0600 Order Number 9228970 The pathophysiology of astrovirus infection in hatchling turkeys Thouvenelle, Marl Lynn, Ph.D. Iowa State University, 1992 UMI 300N.ZeebRd. Ann Arbor, MI 48106 The pathophysiology of astrovirus infection in hatchling turkeys by Mari Lynn Thouvenelle A Dissertation Submitted to the Graduate Faculty in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY Major: Veterinary Pathology Approved; Signature was redacted for privacy. Major Work Signature was redacted for privacy. For the Major Department Signature was redacted for privacy. For the Graduate College Iowa State University Ames, Iowa 1992 11 To my husband, Jerry and daughters, Lauren and Arielle whose support and patience made this work possible iii TABLE OF CONTENTS Page GENERAL INTRODUCTION 1 Explanation of dissertation format 5 LITERATURE REVIEW 6 Historical 6 Identification of Astrovirus 6 Morphology 6 Other identification techniques 6 Virus physicochemical properties 8 Virus classification 8 Virus serotypes 9 Comparative clinical disease and etiologic significance 10 Human 10 Mammal 10 Fowl 12 Comparative pathogenesis in humans and mammals 13 Human 13 Mammal 13 Clinical disease and etiologic significance in turkeys 15 Clinical disease and gross lesions 15 Etiologic significance and epidemiology 15 Pathogenesis in turkeys 16 iv PART I: ASTROVIRUS INFECTION IN HATCHLING TURKEYS: HISTOLOGIC, MORPHOMETRIC AND ULTRASTRUCTURAL FINDINGS ABSTRACT INTRODUCTION MATERIALS AND METHODS Poults and housing Immune electron microscopy Virus inoculum Antiserum Bacteriology Light microscopy and morphometry Experimental design RESULTS Clinical disease and gross lesions Virology and bacteriology Light microscopy and morphometry Electron microscopy DISCUSSION REFERENCES PART II: INTESTINAL DISACCHARIDASE ACTIVITY IN THE SMALL INTESTINE OF ASTROVIRUS INFECTED HATCHLING TURKEYS ABSTRACT INTRODUCTION MATERIALS AND METHODS V Poults and housing 68 Immune electron microscopy 68 Virus inoculum 69 Antiserum 70 Bacteriology 70 Intestinal disaccharidase and protein concentration 70 Experimental design 71 RESULTS 73 DISCUSSION 83 REFERENCES 86 PART III; LECTIN RECEPTOR CHARACTERIZATION OF THE SMALL INTESTINE IN HATCHLING TURKEYS; A PRELIMINARY TRIAL 89 ABSTRACT 91 LITERATURE REVIEW 92 INTRODUCTION 96 MATERIALS AND METHODS 98 Poults and housing 98 Tissue specimens 98 Lectin conjugates 98 Monosaccharides 99 Additional biochemicals 99 Lectin staining 99 Controls 102 Experimental design 103 vi Experimental design 103 RESULTS 104 Group I lectins 104 Group II lectins 104 Group III lectins 104 Group IV lectins 105 Controls 105 DISCUSSION 106 REFERENCES 110 GENERAL DISCUSSION AND SUMMARY 115 LITERATURE CITED 119 ACKNOWLEDGEMENTS 125 APPENDIX; MORPHOMETRIC DATA FROM PART II 126 1 GENERAL INTRODUCTION Astrovirus has been described as single-stranded, non- enveloped RNA virus in man and lambs.Astrovirus virions are 28 to 30 nm in diameter with an ultrastructural star-like morphology. However, only 10 to 20% of the astrovirus particles have the characteristic 5- or 6- pointed white star configuration superimposed on the particle surface when negatively stained. Astrovirus was first reported in association with gastroenteritis in human infants.Subsequently, it has been identified in lambs,*3 calves,** turkeys,2* dogs,*® deer,*® pigs,® cats,^^'^® and mice.^® The virus in turkeys has been incriminated with other enteric viruses as possible causative agents of turkey viral enteritis. Other viruses identified in association with this syndrome include reoviruses, adenoviruses, coronaviruses, 3 groups of rotaviruses, enteroviruses, parvoviruses, and pseudopicornaviruses.^'^®'*® The respective roles of all of these viruses in the pathogenesis of turkey viral enteritis, either individually or in various combinations, remains to be fully explained. Turkey viral enteritis is typically a disease of high morbidity and low mortality, however mortality varies, depending on concomitant health status and environmental conditions affecting the poults. The most common clinical signs are diarrhea and stunting. Necropsy findings include distension of gastrointestinal tract with gas and fluid and dilatation of ceca with yellow frothy feces. There is a high and widespread prevalence of astroviral infection across the USA in turkey flocks (1-5 weeks of age) with enteric disease.Recent studies of the aforementioned enteric viruses in healthy poults and poults with diarrhea of undetermined etiology indicate that single 2 virus isolations are infrequent.^® However, astrovirus-only infections are the most frequent single-virus infection in diseased flocks. Furthermore, astrovirus and rotavirus-like virus (rotavirus group D) are the most frequently identified combination of viruses in diseased flocks from 1-2 weeks to 3- 4 weeks of age. Finally, astroviral infections appear to occur at an earlier age than the other viral infections. Few studies relating to the pathogenicity of astrovirus in animals have been reported. Astrovirus in lambs appears to replicate within the mature villous epithelium and subepithelial macrophages of the small intestine.In a study of small viruses causing acute enteritis in calves, astrovirus alone failed to produce diarrhea although infection was verified.** Astrovirus in combination with rotavirus or breda virus however produces severe watery diarrhea in gnotobiotic calves.The dome epithelial cells of the ileum over Peyer's patches and occasionally enterocytes in the base of adjacent villi, are the sites of astroviral infection in calves infected only with astrovirus.®^ To date, documentation of enteritis associated with astrovirus infections in turkey poults consists of nonspecific gross pathological findings, and significant decreases in both weight gain and intestinal d-xylose absorption.^® There are no reports of concomitant histologic, morphometric or ultrastructural changes in astrovirus infected poults. Measuring intestinal mucosal disaccharidase levels is useful in documenting maldigestion. Complete digestion of oligo- and disaccharides is primarily dependent on the membrane-bound disaccharidases located on the microvilli of enterocytes.4° Maldigestion occurs in gnotobiotic lambs experimentally infected with astrovirus as evidenced by decreased lactase levels.*2 Hatchling poults normally experience a period of maldigestion of fats, proteins and carbohydrates and 3 malabsorption of nutrients within the first 3 to 6 days after hatch.32 Accordingly, the importance of identifying enteric pathogens which augment the innate dysfunction of small intestinal digestion and absorption in the hatchling turkey becomes readily apparent. Natural astroviral infections, as previously mentioned, are characterized by diarrhea and high morbidity with resultant stunting or decreased weight gain. The role astrovirus plays in the pathogenesis of the diarrheic condition and stunting is partially explained by concurrent malabsorption documented by Reynolds et al.^® The possibility of maldigestion occurring coincident to malabsorption in astrovirus infected poults remains unanswered. The
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