Detection of Paramyxovirus, Reovirus and Adenovirus Infection in King Snakes (Lampropeltis triangulum spp.) by Transmission Electron Microscopy and Histopathology Techniques.

Abstract Viruses diverse occur worldwide in reptilian. Paramyxoviruses that infect reptiles belong to Paramyxoviridae family and Ferlavirus genus and are one of the major agents responsible for causing pneumonia in snakes. Reptilian adenovirus has already been documented in various species of snakes, associated with liver, gastrointestinal, respiratory and central nervous system disease. Reptilian orthoreovirus has been demonstrated in several species of reptiles associated with sudden death, central nervous system disorders, skin lesions and pneumonia. In this study 3 kingsnakes (Lampropeltis triangulum spp), from a breeding in Rio de Janeiro, RJ, Brazil, presented a variable clinical picture and death. Feces and organ fragments were processed by the transmission electron microcopy (negative staining) and histopathology (H & E) techniques. By the negative staining, paramyxovirus-like particles, pleomorphic roughly spherical or filamentous, ranging in diameter from 100 to 500 nm, containing internal “herring-borne” nucleocapsid and an outer envelope covered by spikes, were visualized in all samples of the feces and fragments of organs examined. In samples of the small intestine, stomach, pancreas and spleen fragments, adenovirus-like particles, isometric, nonenveloped, containing icosahedral symmetry capsid, measuring 70-90 nm in diameter, were visualized. Reovirus-like particles, isometric, nonenveloped, spherical, characterized as “complete” or “empty”, measuring between 65 and 70 nm in diameter, were also visualized in samples of the feces and small intestine. By the H & E, they were observed in the spleen numerous heterophiles, hypoplastic lymphoid follicles and hyperplastic red pulp. The lung presented hypertrophy and hyperplasia of the alveolar walls, alveoli with cellular debris and mucus; numerous heterophiles and monolymphocytic inflammatory cells. The liver had a marked macro and microgoticular steatosis, with a multifocal presence of nodules in the parenchyma. Monolymphocytic hepatitis and large nuclear basophilic inclusion bodies were also observed in hepatocytes, Kupffer cells and occasionally in endothelial cells. The large intestine presented monolymphocytic enteritis with hyperplasia of enteric lymph nodes and marked proliferation of eosinophils. Some areas showed flattened villi. The skin presented areas with hyperkeratosis, foci with ballooniform degeneration and presence of eosinophilic inclusion corpuscles. In these areas a large number of eosinophils were observed. The kidneys presented monolymphocytic glomerulonephritis. The evaluation of the techniques employed allowed the rapid diagnosis of the viruses in the snakes.

Keywords Transmission electron microscopy, Histopathology, Snakes, Viruses.

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Agriculture Journal- Detection of Paramyxovirus, Reovirus and Adenovirus Infection in King Snakes (Lampropeltis triangulum spp.) by Transmission Electron Microscopy and Histopathology Techniques.

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