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ISOLATION AND CHARACTERIZATION OF A CANINE CORONAVIRUS
Abstract
A virus isolated in cell culture from the feces of dogs suffering from gastroenteritis was identified by physicochemical and morphological properties as a coronavirus. The virus, designated as CCV-18, showed antigenic cross-reactivity with feline coronavirus and transmissible gastroenteritis virus of pigs by immunofluroescence. Both hemagglutination and hemadsorption occurred with different animal erythrocytes, but were standardized using calf erythrocytes. In thin sections of infected A-72 cells, viral particles were observed intracytoplasmically, mainly within vesicles and extracellularly along plasma membranes. By scanning electron microscopy, budding virus-like particles were detected on the surface of virus infected cells. Morphologically, the virus appeared approximately circular in outline and 130 nm in diameter. CCV-18 was characterized by lipid solvents and ('3)H-uridine incorporation experiments, respectively, as containing essential lipid and a RNA genome. The virus was stable at pH values ranging from 2.3 to 10.7, inactivated by heat treatment at 56 C and was not stabilized by 1 M MgCl(,2) at 50 C. The buoyant density of the virions in sucrose density gradients was estimated to be 1.18 gm per cm('3). The CCV-18 nucleic acid was found to consist of a homogenous large-molecular-weight RNA. This viral gene was characterized as a positive-strand RNA because it was susceptible to pancreatic RNase A digestion in high salt, it was polyadenylated, and the RNA extracted from virions was infectious. Low cell-culture passage of CCV-18 produced a very mild infection in susceptible pups with the development of specific serum antibodies. Virus was reisolated from samples of jejumun, ileum, colon, and mesenteric lymph nodes of a killed pup at four days following inoculation. Specific histological features of the small intestine were studied by scanning electron microscopy, immunofluorescence, and light microscopy.
Subject Area
Microbiology
Recommended Citation
GILL, MICHAEL A, "ISOLATION AND CHARACTERIZATION OF A CANINE CORONAVIRUS" (1982). ETD collection for University of Nebraska-Lincoln. AAI8302105.
https://digitalcommons.unl.edu/dissertations/AAI8302105