Adenoviral infection of the renal interstitium of a lamb
| Autor: | S. McConnell, P. F. Cush, J. A. Smyth, B. M. Adair |
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| Rok vydání: | 1990 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty 040301 veterinary sciences Adenoviridae Infections H&E stain Sheep Diseases Spleen Biology Kidney Adenoviridae Inclusion Bodies Viral 0403 veterinary science Immunoenzyme Techniques 03 medical and health sciences medicine Animals Fixative Antiserum Cell Nucleus Bovine adenovirus Sheep General Veterinary Virion 04 agricultural and veterinary sciences medicine.disease Molecular biology Primary and secondary antibodies Immunohistochemistry Small intestine Microscopy Electron 030104 developmental biology medicine.anatomical_structure biology.protein Kidney Diseases |
| Zdroj: | Veterinary pathology. 27(4) |
| ISSN: | 0300-9858 |
| Popis: | Adenoviral infection in sheep is common, and to date, six ovine adenovirus serotypes have been isolated from healthy and diseased sheep.l Adenoviruses serologically indistinguishable from bovine adenoviruses have also been recovered from sheep.' Naturally occumng or experimental adenovirus infections in sheep have caused lesions mainly in the respiratory tract, but adenovirus-induced inclusion bodies have also been reported in hepatocytes of naturally infected lambs.9 This communication describes a novel adenoviral disease in a lamb, characterized by intranuclear inclusion bodies in interstitial cells of kidney and spleen. A 3- to 4 week-old lamb was found dead in its pasture. Necropsy revealed a pale carcass but no other gross abnormalities were seen. Samples of small intestine, mesenteric lymph node, spleen, liver, kidney, and heart were fixed in Brunnel's primary fixative (Laboratory Supplies and Instruments, Antrim, N. Ireland) and embedded in paraffin. Sections were cut at 5 pm and stained with hematoxylin and eosin. Small blocks of fixed kidney were post-fixed, first in cacodylate buffered 4% glutaraldehyde and then in 1% osmium tetroxide, and embedded in epoxy resin. Ultrathin sections were stained with lead citrate and uranyl acetate. Paraffin processed sections of kidney were stained immunocytochemically for adenoviral antigen. An antiserum raised in rabbits against bovine adenovirus serotype 7'O was absorbed with normal ovine kidney and used as primary antibody. Specificity of the absorbed adenovirus antiserum was retested as beforelo and additionally on kidney from lambs dead from other causes. Reagents supplied in a commercially available avidin-biotin-peroxidase complex system (Vectastain Elite Kit, Vector Laboratories, Peterborough, England) were also used. Endogenous peroxidase activity was first blocked using 0.5% hydrogen peroxide in methanol for 20 minutes. After washing them in tap water, the test sections were subjected to 0, 5, 10, 15,20, or 30 minutes protease XIV (Sigma Chemical Company, Poole, Dorset, England) digestion, washed in Tris-buffered saline, and incubated in 2% normal goat serum for 15 minutes. Following overnight incubation with the adenovirus antiserum, sections were washed in Tris-buffered saline, incubated with 0.5% biotinylated goat anti-rabbit IgG for 30 minutes, washed again in Tris-buffered saline, and then incubated with avidin-biotin-peroxidase complex reagent in carbonate buffer (pH 9.4). The substrate (DAB-H,O,) was added for 7 minutes and sections were lightly counterstained with hematoxylin. Formalin-fixed paraffin sections of bovine intestine infected with adenoviruslO were used as positive controls. |
| Databáze: | OpenAIRE |
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