猪δ冠状病毒HeN10株的分离鉴定及致病性研究

doi:10.16431/j.cnki.1671-7236.2021.08.031

Abstract

Abstract: The purpose of this study was to obtain clinical isolates of Porcine deltacoronavirus (PDCoV) and investigate the biological characteristics and pathogenicity of the coronavirus.The swine testicle (ST) cells were used for PDCoV isolation with the PDCoV positive samples obtained clinically from a pig farm in Henan province.The isolated virus was identified by cytopathic effect (CPE) observation, RT-PCR, indirect immunofluorescence (IFA), electron microscopy, and sequence analysis of spike (S) gene, in addition, the pathogenicity was also evaluated using piglets.The PDCoV in the ST cell was successively passaged for 10 passages and confirmed by RT-PCR.The CPE (cell shrinkage, net pulling, fragmentation, and shedding) could be observed in the ST cells from the 4th passage.The isolated virus were identified by IFA, and typical crown shaped viral particles were observed by negative staining in electron microscope, and then the strain of PDCoV was named HeN10.The S gene of HeN10 had the highest sequence identity with China SD strain (99.8%), within the same branch of domestic prevalent strains phylogenetically, such as CHN-JS-2017 and CHN-HeB1-2017.The results of animal test showed that the virus (oral inoculation at a dose of 5×10^6.3 TCID₅₀/piglet) was highly pathogenic by causing watery diarrhea in 10-day-old piglets (5/5).Overall, the PDCoV HeN10 strain was successfully isolated, it showed high sequence identity with domestic prevalent strains such as SD, and also showed high pathogenicity in piglets, which providing a foundation for the development of diagnostic methods and vaccines.

Key words: Porcine deltacoronavirus (PDCoV); isolation and identification; sequence analysis; pathogenicity

CLC Number:

S852.65⁺1

WANG Zhiyan, SUN Yujie, XU Xin, HUANG Baicheng, TIAN Kegong. Isolation,Identification and Pathogenicity of Porcine Deltacoronavirus Strain HeN10[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2982-2989.

References

[1] WANG L,HAYES J,SARVER C,et al.Porcine deltacoronavirus:Histological lesions and genetic characterization[J].Archives of Virology,2016,161(1):171-175.
[2] MARTHALER D,RAYMOND L,JIANG Y,et al.Rapid detection,complete genome sequencing,and phylogenetic analysis of Porcine deltacoronavirus[J].Emerging Infectious Diseases,2014,20(8):1347-1350.
[3] LI G,CHEN Q,HARMON K M,et al.Full-length genome sequence of Porcine deltacoronavirus strain USA/IA/2014/8734[J].Genome Announcements,2014,2(2):e00278-14.
[4] MA Y,ZHANG Y,LIANG X,et al.Origin,evolution,and virulence of Porcine deltacoronaviruses in the United States[J].mBio,2015,6(2):e00064.
[5] JUNG K,HU H,SAIF L J.Porcine deltacoronavirus infection:Etiology,cell culture for virus isolation and propagation,molecular epidemiology and pathogenesis[J].Virus Research,2016,226:50-59.
[6] WOO P C,LAU S K,LAM C S,et al.Discovery of seven novel mammalian and avian coronaviruses in the genus Deltacoronavirus supports bat coronaviruses as the gene source of Alphacoronavirus and Betacoronavirus and avian coronaviruses as the gene source of Gammacoronavirus and Deltacoronavirus[J].Journal of Virology,2012,86(7):3995-4008.
[7] WANG L,BYRUM B,ZHANG Y.Detection and genetic characterization of deltacoronavirus in pigs,Ohio,USA,2014[J].Emerging Infectious Diseases,2014,20(7):1227-1230.
[8] JANETANAKIT T,LUMYAI M,BUNPAPONG N,et al.Porcine deltacoronavirus,Thailand,2015[J].Emerging Infectious Diseases,2016,22(4):757-759.
[9] LEE J H,CHUNG H C,NGUYEN V G,et al.Detection and phylogenetic analysis of Porcine deltacoronavirus in Korean swine farms,2015[J].Transboundary and Emerging Diseases,2016,63(3):248-252.
[10] LORSIRIGOOL A,SAENG-CHUTO K,TEMEE-YASEN G,et al.The first detection and full-length genome sequence of Porcine deltacoronavirus isolated in Lao PDR[J].Archives of Virology,2016,161(10):2909-2911.
[11] SUZUKI T,SHIBAHARA T,IMAI N,et al.Genetic characterization and pathogenicity of Japanese Porcine deltacoronavirus[J].Infection,Genetics and Evolution,2018,61:176-182.
[12] SONG D,ZHOU X,PENG Q,et al.Newly emerged Porcine deltacoronavirus associated with diarrhoea in swine in China:Identification,prevalence and full-length genome sequence analysis[J].Transboundary and Emerging Diseases,2015,62(6):575-580.
[13] MAI K,FENG J,CHEN G,et al.The detection and phylogenetic analysis of Porcine deltacoronavirus from Guangdong province in Southern China[J].Transboundary and Emerging Diseases,2018,65(1):166-173.
[14] ZHANG Y,CHENG Y,XING G,et al.Detection and spike gene characterization in Porcine deltacoronavirus in China during 2016-2018[J].Infection,Genetics and Evolution,2019,73:151-158.
[15] ZHANG H,LIANG Q,LI B,et al.Prevalence,phylogenetic and evolutionary analysis of Porcine deltacoronavirus in Henan province,China[J].Preventive Veterinary Medicine,2019,166:8-15.
[16] DONG N,FANG L,ZENG S,et al.Porcine deltacoronavirus in Mainland China[J].Emerging Infectious Diseases,2015,21(12):2254-2255.
[17] MARTHALER D,JIANG Y,COLLINS J,et al.Complete genome sequence of strain SDCV/USA/Illinois121/2014,a Porcine deltacoronavirus from the United States[J].Genome Announcements,2014,2(2):e00218-14.
[18] CHEN Q,GAUGER P,STAFNE M,et al.Pathogenicity and pathogenesis of a United States Porcine deltacoronavirus cell culture isolate in 5-day-old neonatal piglets[J].Virology,2015,482:51-59.
[19] LIU B J,ZUO Y Z,GU W Y,et al.Isolation and phylogenetic analysis of Porcine deltacoronavirus from pigs with diarrhoea in Hebei province,China[J].Transboundary and Emerging Diseases,2018,65(3):874-882.
[20] 陈建飞,王潇博,焦贺勋,等.国内首株猪德尔塔冠状病毒(Porcine deltacoronavirus)的分离鉴定[J].中国预防兽医学报,2016,38(3):171-174. CHEN J F,WANG X B,JIAO H X,et al.Isolation and identification of the first Porcine deltacoronavirus strain in China[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(3):171-174.(in Chinese)
[21] DONG N,FANG L,YANG H,et al.Isolation,genomic characterization,and pathogenicity of a Chinese Porcine deltacoronavirus strain CHN-HN-2014[J].Veterinary Microbiology,2016,196:98-106.
[22] 张坤,何启盖.猪流行性腹泻病毒、猪传染性胃肠炎病毒和猪A群轮状病毒多重RT-PCR检测方法的建立及临床应用[J].畜牧兽医学报,2010,41(8):1001-1005. ZHANG K,HE Q G.Establishment and clinical application of a multiplex reverse transcription-PCR for detection of Porcine epidemic diarrhea virus,porcine Transmissible gastroenteritis virus and Porcine group A rotavirus[J].Acta Veterinaria et Zootechnica Sinica,2010,41(8):1001-1005.(in Chinese)
[23] HU H,JUNG K,VLASOVA A N,et al.Isolation and characterization of Porcine deltacoronavirus from pigs with diarrhea in the United States[J].Journal of Clinical Microbiology,2015,53(5):1537-1548.
[24] SHIRATO K,MATSUYAMA S,UJIKE M,et al.Role of proteases in the release of Porcine epidemic diarrhea virus from infected cells[J].Journal of Virology,2011,85(15):7872-7880.
[25] WICHT O,LI W,WILLEMS L,et al.Proteolytic activation of the Porcine epidemic diarrhea coronavirus spike fusion protein by trypsin in cell culture[J].Journal of Virology,2014,88(14):7952-7961.
[26] GRAHAM R L,BARIC R S.Recombination,reservoirs,and the modular spike:Mechanisms of coronavirus cross-species transmission[J].Journal of Virology,2010,84(7):3134-3146.
[27] HODGES K,GILL R.Infectious diarrhea:Cellular and molecular mechanisms[J].Gut Microbes,2010,1(1):4-21.
[28] JUNG K,HU H,EYERLY B,et al.Pathogenicity of 2 Porcine deltacoronavirus strains in gnotobiotic pigs[J].Emerging Infectious Diseases,2015,21(4):650-654.
[29] XU Z,ZHONG H,ZHOU Q,et al.A highly pathogenic strain of Porcine deltacoronavirus caused watery diarrhea in newborn piglets[J].Virologica Sinica,2018,33(2):131-141.
[30] ZHANG M J,LIU D J,LIU X L,et al.Genomic characterization and pathogenicity of Porcine deltacoronavirus strain CHN-HG-2017 from China[J].Archives of Virology,2019,164(2):413-425.
[31] WU J L,MAI K J,LI D,et al.Expression profile analysis of 5-day-old neonatal piglets infected with Porcine deltacoronavirus[J].BMC Veterinary Research,2019,15(1):117.

Isolation,Identification and Pathogenicity of Porcine Deltacoronavirus Strain HeN10

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]	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.
[3]	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.
[4]	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.
[5]	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.
[6]	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.
[7]	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.
[8]	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.
[9]	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.
[10]	WEI Jin, WANG Jia, ZHAO Haoran, LIU Bo, LIU Yuanjie, YAO Wensheng, LIU Yan, MA Xin. Isolation, Identification and Antimicrobial Susceptibility of Mycoplasma gallisepticum from Different Areas [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 368-376.
[11]	YANG Fumei, YANG Linfu, ZENG Cheng, YANG Zhi, WANG Kang, DUAN Gang, DAI Feiyan. Isolation,Identification and Biological Characterization Analysis of a Strain of Carnobacterium maltaromaticum from Barking Deer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 398-407.
[12]	ZHANG Wei, WU Hua, YIN Yajie, LI Songli, HOU Shaohua. Isolation and Identification of Novel Duck Reovirus and Its σC Gene Sequence Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3520-3529.
[13]	XI Jing, YI Jihai, WANG Yueli, XU Zhenyu, LI Peidong, ZHANG Jiangwei, CHEN Chuangfu. Isolation, Identification and Genotype Analysis of Group A Bovine Rotavirus G10 Strain XJ-2022 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3530-3538.
[14]	KANG Yuhui, ZHOU Hongshan, JIN Wenjie, QIN Aijian. Isolation, Identification and Pathogenicity of a Strain of Plesiomonas shigelloides from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3676-3682.
[15]	SHAO Mingzhu, ZHAO Yunqing, LI Songli, HOU Shaohua, ZHANG Yanying. Isolation and Identification of Escherichia coli from Dairy Cows in Hebei Province and Its Pathogenicity and Antibacterial Effect of Chinese Herbal Medicine in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3683-3691.