猪流行性腹泻病毒SD201604株的分离鉴定及致病性研究

doi:10.16431/j.cnki.1671-7236.2018.11.030

Abstract

Abstract:

In order to obtain porcine epidemic diarrhea virus (PEDV) isolate and study its pathogenicity,virus isolation was conducted in Vero cells through inoculating diarrhea sample collected from a pig farm in Shandong province.The strain was identified by cytopathogenic effect (CPE),IFA,electron microscope observation and RT-PCR methods,and the S gene sequence of virus was analyzed.The 3-day-old piglets were inoculated orally with 10^3.5 TCID₅₀/mL virus and observed for clinical symptoms and pathological changes.The 3-day-old piglets were inoculated orally with 3 mL different titers of virus.The mortality of piglets in each group was calculated to determine the minimum lethal dose.The results indicated that a strain of PEDV was successfully isolated in the cell culture and designated PEDV SD201604.The strain replicated well in Vero cells and could cause obvious CPE.The infectious titer of the 6th passage of PEDV SD201604 was 10^3.5 TCID₅₀/mL.The strain was detected to be PEDV positive by IFA and RT-PCR methods.Electron microscope observation showed that the size of the virus particle was about 100 nm in diameter,with an obvious envelope and spikes,which was the typical morphological feature of PEDV.Then the virus was identified as a prevalent PEDV variant of China through the analysis of S gene sequence.In the animal pathogenicity experiment,five 3-day-old piglets inoculated orally with 10^3.5 TCID₅₀/mL of PEDV SD201604 strain all showed typical PEDV clinical symptoms and pathological lesions,and 3 of them died in the test.The minimum lethal dose (MLD) experiment showed that piglets all died when they were orally inoculated with 3 mL of virus with a virus content of 10^4.5 TCID₅₀/mL.Together,the experimental results provided references for PEDV isolation,identification and biological characteristics research.

Key words: porcine epidemic diarrhea virus (PEDV); isolation; identification; pathogenicity

CLC Number:

S852.65

WU Bai, SU Weiwei, JU Decai, XIA Mingqi, ZHANG Shucheng, WANG Wei. Isolation, Identification and Pathogenicity Evaluation of Porcine Epidemic Diarrhea Virus SD201604 Strain[J]. , 2018, 45(11): 3229-3236.

References

[1] JEFFREY J Z,LOCKE A K,ALEJANDRO R,et al.Diseases of Swine[M].10th Edition.Wiley-Blackwell,2014.
[2] BI J,ZENG S L,XIAO S B,et al.Complete genome sequence of porcine epidemic diarrhea virus strain AJ1102 isolated from a suckling piglet with acute diarrhea in China[J].Journal of Virology,2012,86(19):10910-10911.
[3] BENJAMIN W N,GABRIELLA K,ANDREAS H K,et al.A structural analysis of M protein in coronavirus assembly and morphology[J].Journal of Structural Biology,2011,174(1):11-22.
[4] MA Y M,ZHANG Y,LIANG X Y,et al.Origin,evolution,and virulence of porcine deltacoronaviruses in the United States[J].MBio,2015,6(2):1-13.
[5] CHEN J F,WANG C B,SHI H Y,et al.Molecular epidemiology of porcine epidemic diarrhea virus in China[J].Archives of Virology,2010,155(2):1471-1476.
[6] ZHANG H W,XIA M Q,JU D C,et al.Isolation,molecular characterization and an artificial infection model for a variant porcine epidemic diarrhea virus strain from Jiangsu province,China[J].Archives of Virology,2017,162(12):3611-3618.
[7] 孙宏伟,王泽岩,任少敏,等.猪主要腹泻病的发展机制综述[J].中国动物检疫,2016,33(3):63-70. SUN H W,WANG Z Y,REN S M,et al.Summary on the pathogenesis of porcine primary diarrhea diseases[J].China Animal Health Inspection,2016,33(3):63-70.(in Chinese)
[8] 王楷宬,王玉梅,李知新,等.宁夏某猪场猪流行性腹泻紧急流行病学调查[J].中国动物检疫,2018,35(4):17-20. WANG K C,WANG Y M,LI Z X,et al.Emergency epidemiological investigation on porcine epidemic diarrhea in a swine farm of Ningxia[J].China Animal Health Inspection,2018,35(4):17-20.(in Chinese)
[9] REED L J,MUENCH H.A simple method for estimating fifty percent endpoints[J].American Journal of Hygiene,1938,27(3):493-497.
[10] MADSON D M,ARRUDA P H E,MAGSTADT D R,et al.Characterization of porcine epidemic diarrhea virus isolate US/Iowa/18984/2013 infection in 1-day-old cesarean-derived colostrum-deprived piglets[J].Veterinary Pathology,2016,53(1):44-52.
[11] STEPHANIE N L,FRANCINE C P,KELLY M L,et al.Lactogenic immunity and vaccines for porcine epidemic diarrhea virus (PEDV):Historical and current concepts[J].Virus Research,2016,226(5):93-107.
[12] WANG J,HU G D,LIN Z,et al.Characteristic and functional analysis of a newly established porcine small intestinal epithelial cell line[J].PLoS One,2014,9(10):e110916.
[13] 朱庆贺,苗艳,张鹏宇,等.猪流行性腹泻病毒细胞受体研究进展[J].动物医学进展,2016,37(9):95-98. ZHU Q H,MIAO Y,ZHANG P Y,et al.Advance in cellular receptor of porcine diarrhea virus[J].Progress in Veterinary Medicine,2016,37(9):95-98.(in Chinese)
[14] LAWRENCE P K,BUMGARDNER E,BEY R F,et al.Genome sequences of porcine epidemic diarrhea virus:In vivo and in vitro phenotypes[J].Genome Announc,2014,2(3):1-2.
[15] LEE D K,PARK C K,KIM S H,et al.Heterogeneity in spike protein genes of porcine epidemic diarrhea viruses isolated in Korea[J].Virus Research,2010,149(2):175-182.
[16] CHANG Y C,KAO C F,CHANG C Y,et al.Evaluation and comparison of the pathogenicity and host immune responses induced by a G2b Taiwan porcine epidemic diarrhea virus (strain Pintung 52) and its highly cell-culture passaged strain in conventional 5-week-old pigs[J].Viruses,2017,9(5):121.
[17] 秦毅斌,卢冰霞,何颖,等.猪流行性腹泻病毒CH/GX/2015/750A株的分离鉴定及全基因组序列分析[J].中国畜牧兽医,2018,45(1):178-188. QIN Y B,LU B X,HE Y,et al.Isolation,identification and complete genome sequence analysis of porcine epidemic diarrhea virus CH/GX/2015/750A[J].China Animal Husbandry & Veterinary Medicine,2018,45(1):178-188.(in Chinese)
[18] GAO Y Y,KOU Q W,GE X N,et al.Phylogenetic analysis of porcine epidemic diarrhea virus field strain prevailing recently in China[J].Archives of Virology,2013,158(3):711-715.
[19] 刘政龙,王金良,沈志强,等.猪流行性腹泻病毒研究进展[J].中国畜牧兽医,2015,42(9):2506-2511. LIU Z L,WANG J L,SHEN Z Q,et al.Research progress on porcine epidemic diarrhea virus[J].China Animal Husbandry & Veterinary Medicine,2015,42(9):2506-2511.(in Chinese)
[20] 牛俊超,黄复深,胡佩佩.猪流行性腹泻研究进展[J].动物医学进展,2014,35(10):107-110. NIU J C,HUANG F S,HU P P.Progress on porcine epidemic diarrhea[J].Progress in Veterinary Medicine,2014,35(10):107-110.(in Chinese)
[21] 代洪波,岳丰雄,徐宏军,等.猪流行性腹泻病毒分离鉴定及其灭活疫苗免疫保护效果研究[J].动物医学进展,2017,38(1):50-55. DAI H B,YUE F X,XU H J,et al.Isolation and identification of porcine epidemic diarrhea virus epidemic strain and study on its immune protection efficacy of inactivated vaccine[J].Progress in Veterinary Medicine,2017,38(1):50-55.(in Chinese)
[22] KNIPPING K,MCNEAL M M,CRIENEN A,et al.A gastrointestinal rotavirus infection mouse model for immune modulation studies[J].Journal of Virology,2011,8(3):1-11.
[23] 朱记平,冯小婷.不同PED疫苗防控猪流行性腹泻的临床效果[J].黑龙江畜牧兽医,2017,9:178-180. ZHU J P,FENG X T.The clinical effect of different PED vaccine against porcine epidemic diarrhea[J].Heilongjiang Animal Science and Veterinary Medicine,2017,9:178-180.(in Chinese)

Isolation, Identification and Pathogenicity Evaluation of Porcine Epidemic Diarrhea Virus SD201604 Strain

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LU Yukui, LIU Jiaqi, ZHONG Jiacheng, CHEN Jidang, ZHU Wanjun, ZHANG Yishan, ZHANG Jipei. Isolation,Identification and Pathogenicity Analysis of Avibacterium paragallinarum from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2562-2572.
[2]	PENG Yangxiang, YANG Zhenbiao, YAN Kuiyou, WANG Han, HU De, WANG Zun, LIU Ning, ZHAO Liansheng. From Artificial to Intelligent:Research Progress of Individual Idendification Technology for Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1855-1866.
[3]	LIU Caixia, SHI Weidi, KOU Lijun, MA Xun, WANG Jing, GAO Shengjie, LYU Shuangfei, CHEN Xuke, ZENG Dongdong, KONG Cuilian, REN Huijie. Regulation of Transcription of Key Virulence Factors of Listeria monocytogenes by LIPI-4 EⅡC [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1340-1351.
[4]	LI Zhijie, ZHAO Ying, MA Xin, WANG Meng, YU Sijiu, WANG Libin, PAN Yangyang. Research Progress on Isolation Methods and Main Reproductive Functions of Exosomes from Animal Follicular Fluid [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1480-1488.
[5]	TIAN Changqing, ZHANG Kunzhong, QI Yumei, SHI Wenjing, DONG Zhijie, ZHANG Haohao, HE Zengwen, ZHI Ji, ZHAO Xuehui, CHONG Qian, XUE Huiwen, GOU Huitian. Isolation,Identification and Biological Characteristics Analysis of a Strain of Listeria monocytogenes from Environment [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1543-1555.
[6]	HUO Mingkai, GUAN Feihu, QIU Runhui, WEI Chunyan, YU Haitao, ZHU Jiale, ZHANG Hui. Study on the Mechanism of DDX1 Inhibiting Porcine Epidemic Diarrhea Virus Replication [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1556-1566.
[7]	LIU Jiaxing, HAN Xueying, ZHANG Xinru, DENG Jiaxin, YAO Lunguang, LIU Yangkun. Preparation and Immunogenicity Analysis of Ferritin Nanoparticles Carrying Antigenic Epitopes of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1567-1574.
[8]	WANG Lucai, SALANG Wenzhu, ZHANG Huanrong. Isolation,Identification and Drug Resistance and Pathogenicity Analysis of Salmonella Enteritidis from Avian [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1663-1674.
[9]	CAO Yingying, LIANG Liang, ZENG Yirong, ZHONG Qinghua, YUAN Xiaofang, TANG Haibo, LENG Jing. Isolation,Identification and Biological Characteristics of Salmonella from Tupaia belangeri chinensis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1718-1728.
[10]	LIU Kexin, WANG Chao, ZHANG Ao, LIU Jiali, TAN Bin, ZHANG Shuqin. Isolation,Identification and Genome Sequence Analysis of Bovine Viral Diarrhea Virus LN-1 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1150-1159.
[11]	CHEN Dong, ZHANG Yanhao, ZHANG Yang, LI Sen, ZHOU Zutao, LI Zili. Isolation,Identification and Drug Sensitivity of 3 Strains of Riemerella anatipestifer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1250-1258.
[12]	WANG Bingyi, MA Hongcai, ZOU Minghao, FAN Shijie, YUAN Zhenjie, Labaciren, DONG Hailong, ZENG Jiangyong. Isolation,Identification and Drug Resistance Analysis of Streptococcus from Yak [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 745-753.
[13]	XIAO Yangyang, LI Ruirui, MA Zhongchen, TANG Tian, WANG Na, CHEN Chuangfu, ZHENG Wei, WANG Yong, WANG Pengyan. Isolation,Identification and Drug Sensitivity Analysis of Mycoplasma bovis in Shihezi Region of Xinjiang [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 754-763.
[14]	ZHONG Huachen, WANG Lifang, GUO Chenyang, LIU Jialin, SONG Jie. Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 817-826.
[15]	SUN Yu, ZHANG Hao, LI Changyang, TAO Yanzi, WANG Qiuju. Isolation,Identification and Biological Characteristics of Antioxidant Bacillus from Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 359-367.