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本研究旨在观察大鲵在大鲵蛙病毒(Chinese giant salamander ranavirus,CGSRV)感染过程中组织的动态病理损伤,同时定量检测CGSRV在组织中的动态分布。对大鲵腹腔注射1.0×106.5 TCID50/m L的CGSRV进行人工感染,在感染的0d,3d,5d,7d,9d,13d,16d随机采集3尾,取肝、肾、脾、肺、肠、皮肤、肌肉、脑、心脏和胃等组织,石蜡切片和HE染色对CGSRV感染大鲵的病理损伤过程进行观察,采用SYBR Green q PCR技术对病毒在组织中的动态分布进行定量研究。组织病理结果表明,CGSRV感染导致大鲵多组织器官损伤,其中肝、脾、肾和皮肤肌肉病变严重,为损伤靶器官,且在一些损伤的细胞内见嗜碱性或嗜酸性包涵体。感染后3d肝细胞与肾小管上皮细胞变性,肾间以及肝小叶中央静脉周围淋巴细胞和嗜酸性粒细胞浸润;感染后5~7d实质性器官变性、坏死,炎症加重,胃肠道呈卡他性炎。感染后9d表皮细胞坏死、脱落,肌纤维变性、坏死。感染后13~16d肝出现广泛变性、坏死与炎症,脾淋巴细胞数量显著减少,肾小球渗出性-坏死性炎,皮肤肌肉呈出血性坏死性炎和实质性心肌炎。SYBR Green qPCR结果显示,整个感染进程中各组织CGSRV含量呈上升趋势,不同组织中病毒量为2.36×103~1.84×109 copy/mg组织,其中肺、肠、肝、脾、肾和皮肤肌肉含量高,表明CGSRV具有广泛的组织分布特征,但肝、脾、肾、皮肤肌肉为其复制和损伤的靶器官,且病毒分布量与病理损伤程度呈正相关。
The aim of this study was to observe the dynamic pathological changes of the tissue during the infection of Chinese giant salamander ranavirus (CGSRV) and to quantitatively detect the dynamic distribution of CGSRV in the tissues. CGSRV (1.0 × 106.5 TCID50 / m L) was intraperitoneally injected into the scallop to induce infection. Three pigs were randomly collected on day 0, day 3, day 5, day 7, day 9, day 13, and day 16 of infection. Liver, kidney, spleen, lung, , Muscle, brain, heart and stomach tissues, paraffin sections and HE staining were used to observe the pathological damage of CGSRV infection. The SYBR Green q PCR technique was used to quantitatively study the dynamic distribution of the virus in the tissues. Histopathological results showed that CGSRV infection caused multiple tissue organ damage in catsup, and liver, spleen, kidney and skin muscle lesions were serious, which was the target organ damage. In some injured cells, basophilic or eosinophilic inclusion bodies were found. After infection, hepatocytes and renal tubular epithelial cells degenerated, interrenal and hepatic lobules around the central venous lymphocytes and eosinophils infiltration; 5 to 7 days after infection, substantial organ degeneration, necrosis, increased inflammation, catarrhal gastrointestinal tract inflammation. Epidermal cells necrosis, shedding, myofibrosis and necrosis 9 days after infection. 13 to 16 days after infection, the liver showed extensive degeneration, necrosis and inflammation, a significant decrease in the number of splenic lymphocytes, glomerular exudative - necrotizing inflammation, hemorrhagic necrotic inflammation of the skin and muscle, and substantial myocarditis. The results of SYBR Green qPCR showed that the contents of CGSRV in different tissues increased from 2.36 × 103 to 1.84 × 109 copies / mg in different tissues, including lung, intestine, liver, spleen, kidney and skin High, indicating that CGSRV has a wide range of tissue distribution characteristics, but liver, spleen, kidney, skin and muscle as its target organ for replication and damage, and the virus distribution and the degree of pathological damage was positively correlated.