2019年云南省景洪市哨兵动物多种虫媒病毒的分离鉴定

doi:10.16431/j.cnki.1671-7236.2020.12.037

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (12): 4127-4137.doi: 10.16431/j.cnki.1671-7236.2020.12.037

2019年云南省景洪市哨兵动物多种虫媒病毒的分离鉴定

杨振兴¹, 寇美玲¹, 廖德芳¹, 高翔², 胡忠燕², 李占鸿¹, 谢佳芮¹, 李华春¹, 杨恒¹

1. 云南省畜牧兽医科学院云南省热带亚热带动物病毒病重点实验室, 昆明 650224;
2. 景洪市动物疫病预防控制中心, 景洪 666100

收稿日期:2020-05-12 出版日期:2020-12-20 发布日期:2020-12-18
通讯作者: 李华春, 杨恒 E-mail:Li_huachun@hotmail.com;yangheng2008.cool@163.com
作者简介:杨振兴(1986-),男,云南昆明人,硕士,助理研究员,研究方向:牛羊虫媒病毒,E-mail:13759463631@163.com;寇美玲(1985-),女,山西太原人,硕士,助理研究员,研究方向:牛羊虫媒病毒,E-mail:617184975@qq.com
基金资助:
国家重点研发计划（2016YFD0500908）；国家重点研发计划（2017YFC1200505）；国家公益性行业（农业）科研专项（201303035）；云南省中青年学术和技术带头人后备人才培养项目（2017HB055）

Isolation and Identification of Arbovirus from the Sentinel Herds in Jinghong City,Yunnan Province,China in 2019

YANG Zhenxing¹, KOU Meiling¹, LIAO Defang¹, GAO Xiang², HU Zhongyan², LI Zhanhong¹, XIE Jiarui¹, LI Huachun¹, YANG Heng¹

1. Yunnan Tropical and Subtropical Animal Virus Disease Laboratory, Yunnan Veterinary and Animal Science Institute, Kunming 650224, China;
2. Animal Epidemic Prevention and Control Center of Jinghong 666100, China

Received:2020-05-12 Online:2020-12-20 Published:2020-12-18

摘要/Abstract

摘要： 本研究旨在了解云南省景洪市虫媒病毒的流行情况。2019年在景洪市勐罕镇设立了3头哨兵动物牛，定期采血进行虫媒病毒的分离与鉴定，共获得7株病毒分离物。经病毒核酸的RT-PCR鉴定，分离到2株血清型分别为6型和7型流行性出血热病毒（epizootic haemorrhagic disease virus，EHDV），2株血清型分别为4型和5型的蓝舌病病毒（bluetongue virus，BTV），1株帕利亚血清群病毒（palyam serogroup virus，PALV）中的D’Aguilar virus（DAV）血清型病毒和2株未鉴定出的环状病毒。经病毒Seg-2、Seg-3序列ORF区的比对和进化分析显示，7株病毒的地域型均为Eastern型，与日本、澳大利亚和印度毒株具有最近的亲缘关系。3头哨兵动物的血液和血清，经病毒核酸及血清中和试验检测，证明3头动物均被相应的病毒感染。动物感染病毒后，血清中的特异性抗体迅速上升，3~4周后达最高点并能够在该水平维持较长时间，而血液中病毒核酸含量2~4周到达最高点后则呈迅速下降趋势。本研究报道了景洪虫媒病毒的分离、毒株序列特征以及在动物上的感染特性，研究结果为进一步了解当地的牛虫媒病毒提供数据支撑，同时3头牛分离获得7株病毒，提示当地可能还存在更多种类的虫媒病毒。

关键词: 蓝舌病病毒(BTV); 流行性出血病病毒(EHDV); 帕利亚血清群病毒(PALV); 病毒分离鉴定

Abstract: The purpose of this study was to understand the prevalence of arboviruses in Jinghong city,Yunnan province.Three sentinel cattle were set up in Menghan township of Jinghong city in 2019,blood samples were collected regularly for isolation and identification of arboviruses,a total of 7 virus isolates were obtained.Viral nucleic acid were identification by RT-PCR,and the results showed that two strains of epidemic hemorrhagic fever virus (EHDV) with serotypes of 6 and 7,two strains of bluetongue virus (BTV) with serotypes 4 and 5,D’Aguilar virus (DAV) serotype in one strain of PALV and two unidentified circoviruses were isolated,respectively.The ORF region of virus Seg-2 and Seg-3 sequences was compared and analyzed,all the 7 regional strains of the virus were Eastern,which was most closely related to strains in Japan,Australia and India.The blood and serum of three sentinel animals were tested by viral nucleic acid and serum neutralization test,which proved that all three animals were infected with the corresponding virus.When animals were infected with the virus,specific antibodies in the serum rise rapidly,it reached peaks 3 to 4 weeks later and could remain at this level for a long time.However,the content of viral nucleic acid in the blood decreased rapidly after reaching the peak from 2 to 4 weeks.The isolation,sequence characteristics and infection characteristics of Jinghong arboviruses in animals were reported in this study,the results provided data support for further understanding of local bovine arboviruses.At the same time,7 strains of virus were isolated from 3 cattle,suggesting that there might be more arboviruses in the area.

Key words: bluetongue virus (BTV); epizootic haemorrhagic disease virus (EHDV); palyam serogroup virus (PALV); virus isolation and identification

中图分类号:

S852.65

杨振兴, 寇美玲, 廖德芳, 高翔, 胡忠燕, 李占鸿, 谢佳芮, 李华春, 杨恒. 2019年云南省景洪市哨兵动物多种虫媒病毒的分离鉴定[J]. 中国畜牧兽医, 2020, 47(12): 4127-4137.

YANG Zhenxing, KOU Meiling, LIAO Defang, GAO Xiang, HU Zhongyan, LI Zhanhong, XIE Jiarui, LI Huachun, YANG Heng. Isolation and Identification of Arbovirus from the Sentinel Herds in Jinghong City,Yunnan Province,China in 2019[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 4127-4137.

参考文献

[1] 梁国栋.虫媒病毒-重要的被忽略的热带传染病病原体[J].中国热带医学,2018,18(1):1-5. LIANG G D.Arbovirus-The very important pathogen of neglected tropical infectious disease in the world[J].China Tropical Medicine,2018,18(1):1-5.(in Chinese)
[2] YANG H,XIAO L,WANG J,et al.Phylogenetic characterization genome segment 2 of bluetongue virus strains belonging to serotypes 5,7 and 24 isolated for the first time in China during 2012 to 2014[J].Transboundary and Emerging Diseases,2016,64(4):1317-1321.
[3] 寇美玲,苗海生,李乐,等.云南江城蓝舌病病毒16型毒株分离鉴定及其L2基因序列分析[J].中国畜牧兽医,2018,45(11):3192-3202. KOU M L,MIAO H S,LI L,et al.Isolation and identification of bluetongue virus type 16 strain from Jiangcheng county in Yunnan province and sequence analysis of its L2 gene[J].China Animal Husbandry & Veterinary Medicine,2018,45(11):3192-3202.(in Chinese)
[4] 杨振兴,寇美玲,李占鸿,等.血清6型流行性出血病病毒在我国云南的首次分离与鉴定[J].中国预防兽医学报,2019,41(11):1113-1119. YANG Z X,KOU M L,LI Z H,et al.Isolation and identification of the first case of epizootic haemorrhagic disease virus serotype 6 strain in Yunan,China for the first time[J].Chinese Journal of Preventive Veterinary Medicine,2019,41(11):1113-1119.(in Chinese)
[5] 寇美玲,杨振兴,李乐,等.云南边境地区流行性出血热病毒血清学监测及血清型鉴定[J].中国畜牧兽医,2019,46(10):3065-3074. KOU M L,YANG Z X,LI L,et al.Serological investigation and serotype identification of epidemic hemorrhagic virus in Yunnan border area[J].China Animal Husbandry & Veterinary Medicine,2019,46(10):3065-3074.(in Chinese)
[6] 杨恒,肖雷,李占鸿,等.2012-2016年中国南方地区帕利亚姆血清群病毒的分离与序列特征分析[J].畜牧兽医学报,2018,49(4):761-770. YANG H,XIAO L,LI Z H,et al.Sequence analysis of palyam serogroup virus isolated in south China from 2012 to 2016[J].Acta Veterinaria et Zootechnica Sinica,2018,49(4):761-770.(in Chinese)
[7] KIRKLAND P D,ZHANG N,HAWKES R A,et al.Studies on the epidemiology of bluetongue virus in China[J].Epidemiology and Infection,2002,128(2):257-263.
[8] MACLACHLAN N J,ZIENTARA S,SAVINI G,et al.Epizootic haemorrhagic disease[J]. Revue Scientifique et Technique (International Office of Epizootics),2015,34(2):341-351.
[9] WHISTLER T,SWANEPOEL R,ERASMUS B J.Characterization of palyam serogroup orbiviruses isolated in South Africa and serologic evidence for their widespread distribution in the country[J].Epidemiology and Infection,1989,102(2):317-324.
[10] ROY P.Bluetongue virus proteins[J].Journal of General Virology,1992,73(12):3051-3064.
[11] ANTHONY S J,MAAN N,MAAN S,et al.Genetic and phylogenetic analysis of the core proteins VP1,VP3,VP4,VP6 and VP7 of epizootic haemorrhagic disease virus (EHDV)[J].Virus Research,2009,154(2):190-199.
[12] ANTHONY S J,MAAN S,MAAN N,et al.Genetic and phylogenetic analysis of the outer-coat proteins VP2 and VP5 of epizootic haemorrhagic disease virus (EHDV):Comparison of genetic and serological data to characterise the EHDV serogroup[J].Virus Research,2009,145(2):200-210.
[13] VELIZAR B,NATALIA G,MARIA J,et al.Characterization of bluetongue virus serotype 28[J].Transboundary and Emerging Diseases,2020,67(1):171-182.
[14] SHIRAFUJI H,KATO T,YAMAKAWA M,et al.Characterization of genome segments 2,3 and 6 of epizootic hemorrhagic disease virus strains isolated in Japan in 1985-2013:Identification of their serotypes and geographical genetic types[J].Infection Genetics and Evolution,2017,53:38-46.
[15] WHISTLER T,SWANEPOEL R.Characterization of potentially foetotropic palyam serogroup orbiviruses isolated in Zimbabwe[J].Journal of General Virology,1988,69(9):2221-2227.
[16] CARPENTER S,WILSON A,MELLOR P S.Culicoides and the emergence of bluetongue virus in Northern Europe[J].Trends in Microbiology,2009,17(4):170-178.
[17] WILSON W C,RUDER M G,KLEMENT E,et al.Genetic characterization of epizootic hemorrhagic disease virus strains isolated from cattle in Israel[J].Journal of General Virology,2015,96(6):1400-1410.
[18] ETHEM M,TEMIZEL,KADIR,et al.Epizootic hemorrhagic disease in cattle,Western Turkey[J].Emerging Infectious Diseases,2009,15(2):317-319.
[19] TATEYAMA S,YAMAGUCHI R,UCHIDA K.An outbreak of congenital hydranencephaly and cerebellar hypoplasia among calves in south Kyusyu,Japan:A pathological study[J].Research in Veterinary Science,1990,49(2):127-131.
[20] 张海林,施华芳,米竹青,等.云南省景洪市虫媒病毒调查分析[J].地方病通报,2000,3:40-44. ZHANG H L,SHI H F,MI Z Q,et al.Investigation of arboviruses in Jinghong city,Yunnan province,China[J].Endemic Diseases Bulletin,2000,3:40-44.(in Chinese)
[21] 唐承军,付士红,张海林,等.云南省西双版纳地区2011年蚊虫及虫媒病毒调查[J].中国媒介生物学及控制杂志,2012,23(5):410-412. TANG C J,FU S H,ZHANG H L,et al.Investigation of mosquitoes and arboviruses in Xishuangbanna,Yunnan province,China in 2011[J].Chinese Journal of Vector Biology and Control,2012,23(5):410-412.(in Chinese)
[22] KATOH K,ASIMENOS G,TOH H.Multiple alignment of DNA sequences with MAFFT[J].Methods in Molecular Biology,2009,537(8):39.
[23] TAMURA K,STECHER G,PETERSON D,et al.MEGA6:Molecular evolutionary genetics analysis version 6.0[J].Molecular Biology and Evolution,2013,30(12):2725-2729.
[24] T18638-2017.蓝舌病诊断技术[S].北京:中国标准出版社,2017. T18638-2017.Diagnostic technique for bluetongue[S].Beijing:Standard Press of China,2017.(in Chinese)
[25] 潘虹,张海林,高阳,等.云南省景洪市2015年蚊虫感染病毒状况调查[J].中国媒介生物学及控制杂志,2018,29(4):331-335. PAN H,ZHANG H L,GAO Y,et al.Identification of Japanese encephalitis viral infections in mosquitoes in Jinghong city,Yunnan province,China[J].Chinese Journal of Vector Biology and Control,2018,29(4):331-335.(in Chinese)
[26] CLAVIJO A,HECKERT R A,DULAC G C,et al.Isolation and identification of bluetongue virus[J].Journal of Virological Methods,2000,87(1):13-23.
[27] 杨振兴,孟锦昕,肖雷,等.流行性出血病病毒(EHDV)血清7型毒株在中国的首次分离与鉴定[J].畜牧兽医学报,2019,50(3):150-158. YANG Z X,MENG J X,XIAO L,et al.Isolation of epizootic haemorrhagic disease virus serotype 7 strain in China for the first time[J].Acta Veterinaria et Zootechnica Sinica,2019,50(3):150-158.(in Chinese)
[28] 李占鸿,肖雷,孟锦昕,等.中国蓝舌病病毒血清5型毒株的分离与鉴定[J].中国兽医科学,2019,49(1):36-43. LI Z H,XIAO L,MENG J X,et al.Isolation and identification of bluetongue virus serotype 5 strain in China[J].Chinese Veterinary Science,2019,49(1):36-43.(in Chinese)
[29] WANG F,LIN J,CHANG J,et al.Isolation,complete genome sequencing,and phylogenetic analysis of the first Chuzan virus in China[J].Virus Genes,2016,52(1):138-141.

2019年云南省景洪市哨兵动物多种虫媒病毒的分离鉴定

Isolation and Identification of Arbovirus from the Sentinel Herds in Jinghong City,Yunnan Province,China in 2019

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 3

编辑推荐

Metrics

本文评价

[1]	李占鸿, 宋子昂, 肖雷, 朱建波, 杨振兴, 李卓然, 廖德芳, 李华春, 杨恒. 流行性出血病病毒VP7蛋白原核表达及其多克隆抗体的制备[J]. 中国畜牧兽医, 2021, 48(4): 1405-1413.
[2]	寇美玲, 苗海生, 李乐, 李楠, 廖德芳, 李华春. 云南江城蓝舌病病毒16型毒株分离鉴定及其L2基因序列分析[J]. 《中国畜牧兽医》, 2018, 45(11): 3192-3202.
[3]	孙建富;郝建伟;荣福龙;姜力;姜丽萍;白宇. 猪繁殖与呼吸综合征病毒的分离与鉴定及 ORF5基因的序列分析[J]. , 2011, 38(9): 138-142.