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Marble spleen disease (MSD) of ring-necked pheasants (Phasianus colchicus L.) was transmitted in the laboratory to pheasants and turkeys (Meleagris gallopavo L.) by oral, colonic, and intravenous routes of inoculation, using cell-free supernatant fluids of splenic suspension from birds with naturally occurring MSD. Gross lesions consisted of large mottled spleens, a lesion more prominent in infected pheasants than in turkeys. The only microscopic lesion and also the criterion of infection were the presence of typical intranuclear inclusions of MSD. Similar to microscopic observations in naturally infected birds was the presence of inclusions in cells of spleen, bone marrow, liver, lung, bursa of Fabricius, and intestine-associated lymphoid tisse (IALT) OF EXPERIMENTALLY INFECTED BIRDS. Ultrastructural examination of splenic cells with intranuclear inclusions from turkey and pheasant with experimentally transmitted disease revealed viral particles and inclusions morphologically indistinguishable from those observed in naturally infected birds. Results of viral isolation procedures in turkey embryo chorioallantoic membrane (CAM) and yolk sac, as well as in turkey embryo fibroblast (TEF) and turkey kidney cell (TKC) cultures, were negative. Serologic assay by agar gel immunodiffusion tests indicated cross-reactivity between splenic MSD antigen and spleinic hemorrhagic enteritis (HE) of turkey antigen. Lines of fusion were formed between the splenic antigen from naturally occurring MSD in pheasants, experimentally transmitted MSD in turkeys, and HE of turkeys, using the homologous serum antibody to each of the 3 antigens.

A purification procedure, using chloroform or fluorocarbon extraction and centrifugation on a cushion of cesium chloride (CsCL), was designed to isolate the causative virus of marble spleen disease. Virus was purified, inoculated into turkeys, and subsequently reisolated by purification from the spleen of inoculated turkeys, thus fulfilling Koch's postulates. Splenic antigen was detected by the agar gel precipitin test, and viral inclusions with viral particles were observed by light and electron microscopy. Results of further studies indicated splenic lymphoreticulum cell hyperplasia was a sensitive indicator of marble spleen disease virus (MSDV) infection. Direct fluorescent antibody staining revealed nuclear fluorescence in virus-infected splenic cells from turkeys inoculated with purified MSDV. With negative stain electron microscopy, MSDV was observed to be 90 nm across with an icosahedral capsid composed of 252 capsomeres. This morphologic feature was consistent with that of an avian adenovirus. Serologic evidence for classification of MSDV as an adenovirus was the cross reaction of MSDV antigen with antiserum to turkey adenovirus serotypes TA-1 and TA-2 in the agar gel precipitin test.

To determine if the rabbit can act as an intermediate host for ascarids from certain carnivorous animals (skunks, pigs, dogs), 12 rabbits were infected with Ascaris columnaris, Ascaris suum, or Toxocara canis. Clinical signs, microscopic lesions, and morphologic features of ascarids with naturally occurring and experimentally induced A columnaris infections were identical. Of the 3 induced parasitic infections, A columnaris caused the most severe lesions in the central nervous system (CNS). It appears that rabbits may potentially act as intermediate hosts for A columnaris.