CP和NCP型牛病毒性腹泻病毒腹腔注射小鼠感染模型的建立  

Establishment of infection model of CP and NCP bovine viral diarrhea virus by intraperitoneal injection in mice

作　　者：吴陈华 王丽娜 岳山[1] 陈楠楠[1] 黄雯静 高丽 付金蕾 刘思雨 赵童 白彤彤 苏思雨 黄江 白彪慧 金鑫 范春玲[1,2] 刘宇 张泽财[1,2] 周玉龙 朱战波[1,2] WU Chen-hua;WANG Li-na;YUE Shan;CHEN Nan-nan;HUANG Wen-jing;GAO Li;FU Jin-lei;LIU Si-yu;ZHAO Tong;BAI Tong-tong;SU Si-yu;HUANG Jiang;BAI Biao-hui;JIN Xin;FAN Chun-ling;LIU Yu;ZHANG Ze-cai;ZHOU Yu-long;ZHU Zhan-bo(Heilongjiang Bayi Agricultural University,Daqiang 163319,China;Heilongjiang Province Cattle Disease Prevention and Control Technology Innovation Center,Daqiang 163319,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]黑龙江省牛病防控技术创新中心,黑龙江大庆163319

出　　处：《中国预防兽医学报》2022年第1期28-33,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金项目(32072896);省重点实验室开放课题(PCBD201705);黑龙江八一农垦大学研究生创新科研项目(YJSCX2019-Y31、YJSC2018-Y26)。

摘　　要：为了建立不同生物型牛病毒性腹泻病毒(BVDV)小鼠的急性感染模型,本研究将0.4 mL 104TCID50/mL致细胞病变型(CP型)BVDV NADL株和非致细胞病变型(NCP型)BVDV NY-1株分别经腹腔注射BALB/c小鼠,观察10 d,分别在感染后第4 d、7 d、10 d每组分别采集3只小鼠的血液、粪便后迫杀,无菌采集其心、肝、脾、肺、肠等组织样品分别进行相应检测。采集血液的血常规检测结果显示,与对照组相比,感染后第4 d、7 d和10 d,CP组和NCP组小鼠的白细胞、淋巴细胞和血小板数量均下降,在第7 d下降最明显;且感染后第7 d、10 d CP组小鼠的上述指标均低于NCP组。各组织、血液和粪便的荧光定量PCR检测结果显示,CP组小鼠在上述3个时间点血液中BVDV的载量均保持较高水平。感染后第7 d CP组和NCP组小鼠各组织中均能检测到病毒,其中血液中的病毒载量最高,对照组小鼠各组织均未检测到相应病毒。免疫组化检测感染后第7 d各组织中BVDV抗原的分布,结果显示,感染后第7 d CP组和NCP组小鼠各组织中均检测到了BVDV抗原,对照组小鼠则均未检测到相应抗原。各组小鼠的组织病理学观察结果显示,CP组小鼠十二指肠肠腺上皮细胞变性坏死脱落、间质疏松水肿和炎性细胞浸润,肝脏肝细胞肿胀、空泡变性和坏死,脾脏间质疏松水肿;NCP组小鼠十二指肠黏膜上皮细胞变性、坏死、脱落,肝脏肝血窦可见淋巴细胞炎性浸润,脾脏淋巴细胞变性和坏死,对照组小鼠各组织均无明显病理变化。本研究结果表明CP型BVDV的致病性强于NCP型BVDV,且建立了两种不同生物型BVDV急性感染的小鼠模型,为BVDV疫苗的研制和致病机理的研究奠定基础。In order to establish a mouse model for acute bovine viral diarrhea virus(BVDV) infection,BALB/c mice were injected intraperitoneally with BVDV NADL strain(CP type) or NY-1 strain(NCP type) at a dose of 0.4 m L containing 104 TCID50/m L,followed by observation for ten days.The blood and feces of 3 mice in each group were collected at 4 days,7 days,and 10 days post-infection(dpi).The heart,liver,spleen,lung,and intestine samples were collected aseptically for the corresponding detection.The blood routine test showed that the number of leukocytes,lymphocytes,and platelets in the CP NADL and NCP NY-1 infected groups decreased.The most significant decrease was at 7 dpi compared to the control group.The above indexes in the NADL infected group were lower than those in NY-1 infected group at 7 dpi and 10 dpi.Real-time quantitative fluorescent reverse transcriptase-PCR(RT-qPCR) detection of tissues,blood and feces showed that the viral load of BVDV in the blood of NADL infected mice maintained a high level at the above three time points.The virus could be detected in all tissues of mice in either NADL or NY-1 infection group at 7 dpi,in which the virus load in blood was the highest.In contrast,no corresponding virus was detected in all tissues of mice in the control group.The distribution of BVDV antigen in different tissues was detected by immunohistochemistry at 7 dpi.The results showed that BVDV antigen was detected in all tissues of NADL and NY-1 infection group,while no BVDV antigen was observed in the control group.The histopathological results showed that duodenal intestinal gland epithelial cells degenerated,necrotic and shed,interstitial edema and inflammatory cell infiltration,liver hepatocyte swelling,vacuolar degeneration and necrosis,and splenic interstitial edema in the NADL infection group;while the epithelial cells of duodenal mucosa degenerated,necrotic and exfoliated,the inflammatory infiltration of lymphocytes in hepatic sinusoids and the degeneration and necrosis of lymphocytes in spleen were observe

关 键 词：牛病毒性腹泻病毒 BALB/C小鼠 腹腔注射 急性感染 模型 

分 类 号：S852.65[农业科学—基础兽医学]