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《Acta Biologiae Experimentalis Sinica》 1986-01
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Deng Zhen-wen Li Yon-bao Sun Hsich-chun and Yao Tseng-hsu (Shanghai Institute of Cell Biology,Academia Sinica) Liu Hong-chang Xiao Qing-lin and Mao Shi-hong (Shanghai College of Agriculture)  
Since last year,many rabbit breedingcolonies in different regions in China hadbeen successively attacked with an acute di-sease,characterized by sudden onset,earlydeath,and usually high mortality rate.Ty-pical clinical symptoms of the diseased ani-mals were dull,no appetite and continuousfever.Morbid anatomy showed haemorrhagein liver,kidney,lung and spleen.Observa-tions made previously on ultrathin sectionsof the haemorrhagic tissue under electronmicroscope indicated that numerous virus-likeparticles were present in the cytoplasm ofthe almost disintegrated cells(Fig 1).In thepresent report,virus isolation and purifica-tion were performed from homogenates of rab-bit haemorrhagic liver and kidney,aimingat characterizing the virus-like particles andascertaining the viral etiology of this infec-tious disease. Purified preparation was obtained throughsucrose density gradient centrifugation.Byelectron microscopic examination,a large nu-mber of nonenveloped virus particles,withan outer diameter 34—37 nm and an innerdiameter about 20 nm was observed.The ou-ter surface of the virus was evenly spacedwith knob-like structures.From both thesize and morphological features,the isolatedvirus is very similar to calicivirus(Fig 2).Nucleic acid extracted with phenol/chloro-form from purified virus preparation afterhaving been digested with proteinase K,wasidentified by paper chromatogram method de-scribed by Buchanan(1951).Purified viruspreparation was also stained with acridine ??orange along with those treated by RNase orDNase for differentiating RNA and DNAvirus,according to the procedure described byHsuing(1973).Both results indicated thatthe virus is of RNA type and may be singlystranded.To confirm the etiological relation-ship,we infected two healthy young rabbitsthrough intramuscular injection of suspensionof the purified virus preparation.Typicalclinical sign,as mentioned above,appearedwithin 24 hours,and haemorrhage in tissuesalso manifested after morbid anatomy.In ad-dition,when sera obtained from a few rab-bits,survived after repeated injections withhomogenate made from the haemorrhagic tis-sues and one boosted injection,were addedto the purified virus suspension,obvious ag-glutination of the viral particles occurred.This result further indicated that the survi-ved rabbits had produced antibodies agaist theisolated RNA virus after immunization.Fromthese findings,together with that similarvirus particles were usually found in thehaemorrhagic tissues from diseased rabbits inthe farm,it is evident that the isolated RNAvirus is the pathogen of the acute infectiousdisease of rabbit.In accordance with theclinical and pathological features,we tenta-tively suggest that this newly isolated virusmay be named haemorrhagic fever virus ofrabbit(Oryctolagus cuniculus domestica),be-longing to picornaviridae.
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