_鸭副粘病毒的分离与初步鉴定

第26卷 第2期 中 国 预 防 兽 医 学 报 Vol.26,No.2 2004年3月 Chinese Journal of Preventive Veterinary Medicine Mar.2004

鸭副粘病毒的分离与初步鉴定

陈少莺,胡奇林,陈仕龙,程晓霞,吴南洋,林锋强,朱小丽,程由铨

(福建农科院畜牧兽医研究所,福建福州,350013)

摘要:从疑为流感的病死鸭肝、脾和肾脏中分离到1株病毒,该病毒能凝集鸽红细胞,并能被康复鸭血清、新城疫标准阳性血清抑制,但不能被EDS、禽流感H5、H7、H9血清抑制;电镜观察见到形态不规则、囊膜表面具有密集纤突的病毒粒子,直径100~200n m。部分生物学特性表明:该分离毒具有较强的毒力,其对鸡胚和番鸭胚的最小致死量的平均致死时间(MDT)为43.6和48.6h;接种鸡胚成纤维细胞和番鸭胚成纤维细胞均能引起细胞病变,且该病变能被康复鸭血清、新城疫标准阳性血清中和,而不能被GPV、MPV、H5、EDS血清所中和。人工感染1日龄雏番鸭和50日龄鸡均可发病并能回收到病毒。上述结果初步表明该分离毒为鸭副粘病毒。

关键词:鸭副粘病毒; 分离; 鉴定

中图分类号:S852.65+9.6 文献标识码:B 文章编号:1008_0589(2004)02_0118_03

Isolation and identification of duck paramyxovirus

CHEN Shao_ying,HU Qi_lin,C HEN Shi_long,CHENG Xiao_xia,WU Nan_yang,LIN Feng_qiang,ZHU Xiao_li,C HENG You_q uan (Ins ti tute of Ani mal Hus bandry and Veterinary M edicine,FuJian Academy of Agricul tural Sci ences,Fuz hou,350013,China) Abstract:One isolated virus was got from the liver,spleen and kidney of ducks wi th suspicions of duck Influenza.The isolate was able to agglu tinate pigeon haemocytes and inhibited by sera of recovered duck and sera agai ns t newcastle disease virus(NDV),But it didn t inhibited by sera of H5、H7、H9、EDS.The isolate possessed irregulated_shape and enveloped wi th lots of peplomeres about100__ 200n m in diameter under electronic_microscopy.The detection of biological property showed that the isolate was very virulen t,its MDT to chicken embryo and Muscovy duck embryo were43.6hrs and48.6hrs;and the isolate propagated in muscovy duck embryo fibroblast (MDEF)and chicken embryo fibroblas t(CE F)and caused to cytopathogenic effect(CPE),and the CPE were neutralized by sera of re covered duck and sera against NDV and didn t neutralized by sera of GPV,MPV,H5,EDS.The one_day_old Muscovy d ucks and50_day_ old chicks inoculated with the isolate showed disease,and the isolate was recovered from the tissues of these ducks and chicks.These re sults suggested that the isolate was duck paramyxovirus.

Key words:duck paramyxoviru; isolation; identification

2002年3月,福州郊县某鸭场饲养的2500多羽半番鸭发生一种以拉白色稀粪、肝出血、脾黄白色坏死点和胰腺出血等为特征的疫病,发病率40%左右,病死率约25%左右,疑为鸭流感,但应用抗菌素、抗流感药物和各种高免抗体均未能控制病情,为确诊该病,故采集病料进行病原分离和鉴定。现将试验结果报告如下。

1 材料和方法

1.1 材 料 番鸭胚、鸡胚、雏番鸭、鸡等均来自非疫区;新城疫标准强毒F48E8株及阳性血清、减蛋综合征阳性血清,均购自中国兽药监察所;禽流

作者简介:陈少莺(1962~),女,副研究员.主要从事畜禽病毒病及免疫防制研究.

收稿日期:2003-05-22感琼扩抗原及抗体、禽流感H5、H7和H9标准HI 抗原及抗体,均购自哈尔滨兽医研究所;鹅细小病毒(GPV)、番鸭细小病毒(MPV)、鸭病毒性肝炎阳性血清系本室自制并保存。

1.2 病毒分离

1.2.1 病料处理:取病鸭的肝、脾、肾,剪碎研磨,以Hank s液制成1 5匀浆,加双抗各1000U, 4 过夜,冻融3次,7000r/min离心10min,取上清供接种材料。

1.2.2 病毒分离:取处理的病料上清液经尿囊腔接种13日龄番鸭胚各3枚,0.1mL/枚,37 孵化,每日观察,弃去24h内的死胚,收取24~96h 的死胚或活胚,置4 过夜后,收获番鸭胚尿囊液,并观察胚胎病变。

1.3 血清学鉴定

1.3.1 血凝价(HA)测定:按常规方法测定鸡胚、