川藏地区牦牛牛病毒性腹泻病毒分子流行病学调查及分离鉴定

doi:10.11843/j.issn.0366-6964.2018.03.018

摘要/Abstract

摘要：

为了解川藏高原地区牦牛牛病毒性腹泻病毒（bovine viral diarrhea virus，BVDV）的感染情况和牦牛BVDV的主要流行亚型及遗传变异情况，采用RT-PCR方法，对四川和西藏部分高原地区采集的149份临床腹泻牦牛粪便开展分子流行病学调查，根据5'-UTR、Npro和E2基因的变异研究该地区牦牛BVDV的遗传变异情况。同时，通过RT-PCR、病毒分离培养和间接免疫荧光检测等方法鉴定牦牛BVDV分离株。结果表明，149份粪便中，BVDV阳性检出率为19.46%[95%置信区间（CI）=13.4%~26.7%]，其中西藏地区牦牛BVDV阳性检出率为27.14%（95% CI=17.2%~39.1%），四川地区牦牛BVDV阳性检出率为12.66%（95% CI=6.2%~22.0%）。随机抽取10份阳性样本测序，根据5'-UTR、Npro和E2基因序列建立系统发育进化树分析，10份样本为BVDV-1型，包括BVDV-1a（n=4）和1d（n=6）亚型。此外，成功分离鉴定出两株致细胞病变型（cytopathic，cp）BVDV，分别属于牦牛BVDV-1a和1d亚型，命名为SWU-Z10和SWU-L8。研究结果提示，川藏部分高原地区牦牛存在BVDV感染，BVDV-1a和1d为牦牛感染的主要亚型。本研究丰富了BVDV的分子流行病学资料，也为后续研制牦牛专用BVDV疫苗提供理论基础。

Abstract:

This study was conducted to investigate the infection status of Bovine viral diarrhea virus (BVDV) and the main epidemic subtypes and genetic variation of BVDV in the Sichuan-Tibet Plateau region. A total of 149 clinical diarrhea yak fecal samples were collected from Sichuan province and Tibet region in 2016, and the genetic variation of BVDV in yak was studied according to the variation of 5'-UTR, Npro and E2. Meanwhile, the BVDV strain of yak was isolated and identified by RT-PCR, cell isolation and indirect immunofluorescence. The results showed that the positive rate of BVDV was 19.46% (95% confidence interval (CI)=13.4% -26.7%) in 149 samples of clinical diarrhea yak feces. The positive rate of BVDV in yak in Tibet region was 27.14% (95% CI=17.2% -39.1%) and 12.66% (95% CI=6.2%-22.0%) in Sichuan province. The phylogenetic tree was established based on the sequences of 5'-UTR, Npro and E2. The results showed that 10 samples belongs to BVDV-1, BVDV-1a subtype (n=4) and 1d (n=6). In addition, two strains of cytopathic (cp) BVDV were identified and identified as subgroups of BVDV-1a and 1d, named SWU-Z10 and SWU-L8, respectively. The results showed that BVDV infection is present in yak in the Sichuan-Tibet Plateau region, and BVDV-1a and 1d are the dominant subtype of yak infection in Sichuan-Tibet Plateau region. The results of the study provide basic data for the comprehensive prevention and control of yak diarrhea in Sichuan-Tibet Plateau region. The data obtained enrich the molecular epidemiological investigation data of BVDV, and provide the theoretical basis for the development of BVDV vaccine for yak.

中图分类号:

S852.659.6

陈新诺, 肖敏, 阮文强, 覃思楠, 岳华, 汤承, 张斌. 川藏地区牦牛牛病毒性腹泻病毒分子流行病学调查及分离鉴定[J]. 畜牧兽医学报, 2018, 49(3): 606-613.

CHEN Xin-nuo, XIAO Min, RUAN Wen-qiang, QIN Si-nan, YUE Hua, TANG Cheng, ZHANG Bin. Molecular Epidemiological Investigation and Isolation of Bovine Viral Diarrhea Virus in Yak in Sichuan-Tibet Plateau Region[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 606-613.

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参考文献

[1] LANYON S R, HILL F I, REICHEL M P, et al. Bovine viral diarrhoea:pathogenesis and diagnosis[J]. Vet J, 2014, 199(2):201-209.
[2] 韩玉霞, 孟露萍, 孙志华, 等. 不同生物型BVDV感染对MDBK细胞类泛素基因转录水平的影响[J]. 动物医学进展, 2015, 36(12):11-17.
HAN Y X, MENG L P, SUN Z H, et al. Effects of different biological-types of Bovine Viral Diarrhea Virus infection on SUMO gene transcription levels in MDBK cells[J]. Progress in Veterinary Medicine, 2015, 36(12):11-17. (in Chinese)
[3] BACHOFEN C, VOGT H R, STALDER H, et al. Persistent infections after natural transmission of bovine viral diarrhoea virus from cattle to goats and among goats[J]. Vet Res, 2013, 44:32.
[4] TAO J, LIAO J H, WANG Y, et al. Bovine viral diarrhea virus (BVDV) infections in pigs[J]. Vet Microbiol, 2013, 165(3-4):185-189.
[5] HELAL M A, OKAMATSU H, TAJIMA M. Bovine viral diarrhea virus infection in a dairy herd with high prevalence of persistently infected calves[J]. Jpn J Vet Res, 2012, 60(2-3):111-117.
[6] PINIOR B, FIRTH C L, RICHTER V, et al. A systematic review of financial and economic assessments of bovine viral diarrhea virus (BVDV) prevention and mitigation activities worldwide[J]. Prev Vet Med, 2017, 137:77-92.
[7] GÍMEZ-ROMERO N, BASURTO-ALCÁNTARA F J, VERDUGO-RODRÍGUEZ A, et al. Genetic diversity of bovine viral diarrhea virus in cattle from Mexico[J]. J Vet Diagn Invest, 2017, 29(3):362-365.
[8] COUVREUR B, LETELLIER C, COLLARD A, et al. Genetic and antigenic variability in bovine viral diarrhea virus (BVDV) isolates from Belgium[J]. Virus Res, 2002, 85:17-28.
[9] YESILBAG K, ALPAY G, BECHER P. Variability and global distribution of Subgenotypes of bovine viral diarrhea virus[J]. Viruses, 2017, 9(6):128.
[10] DENG M L, JI S K, FEI W T, et al. Prevalence study and genetic typing of bovine viral diarrhea virus (BVDV) in four bovine species in China[J]. PLoS One, 2015, 10(4):e0134777.
[11] GIAMMARIOLI M, CEGLIE L, ROSSI E, et al. Increased genetic diversity of BVDV-1:recent findings and implications thereof[J]. Virus Genes, 2015, 50(1):147-151.
[12] ZEMKE J. Characterization of recombinant BVDV-2 vaccine prototypes based on packaged replicons and replication competent deletion mutants[D]. LMU München:Tierärztliche Fakultät, 2010.
[13] 张倩, 陶洁, 张东, 等. 国内新型基因2型牛病毒性腹泻病毒研究进展[J]. 中国动物传染病学报, 2017, 25(1):80-86.
ZHANG Q, TAO J, ZHANG D, et al. Research progress of genotype 2 bovine viral diarrhea virusin China[J]. Chinese Journal of Veterinary Parasitology, 2017, 25(1):80-86. (in Chinese)
[14] 陈新诺, 张朝辉, 徐林, 等. 青藏高原牦牛感染BVDV和BEV的分子流行病学调查[J]. 动物医学进展, 2016, 37(9):35-38.
CHEN X N, ZHANG C H, XU L, et al. Investigation on molecular epidemiology of BVDV and BEV infections in yaks of Qinghai-Tibetan Plateau[J]. Progress in Veterinary Medicine, 2016, 37(9):35-38. (in Chinese)
[15] 何美琳, 张焕容, 王永, 等. 川西北牦牛3种病毒性腹泻病血清学调查[J]. 中国畜牧兽医, 2014, 41(3):248-251.
HE M L, ZHANG H R, WANG Y, et al. Serological survey on three viral diarrhea diseases of yaks in Northwest Sichuan Province[J]. Chinese Journal of Animal Husbandry and Veterinary Medicine, 2014, 41(3):248-251. (in Chinese)
[16] KUTA A, POLAK M P, LARSKA M, et al. Predominance of bovine viral diarrhea virus 1b and 1d subtypes during eight years of survey in Poland[J]. Vet Microbiol, 2013, 166(3-4):639-644.
[17] GONG X W, LIU L H, ZHENG F Y, et al. Molecular investigation of bovine viral diarrhea virus infection in yaks (Bos gruniens) from Qinghai, China[J]. Virol J, 2014, 11:29.
[18] 拜彬强, 郝力壮, 柴沙驼, 等. 牦牛肉品质特性研究进展[J]. 食品科学, 2014, 35(17):290-296.
BAI B Q, HAO L Z, CHAI S T, et al. Progress in understanding meat quality characteristics of yak[J]. Food Science, 2014, 35(17):290-296. (in Chinese)
[19] 李齐发, 赵兴波, 刘红林, 等. 牦牛分类地位研究概述[J]. 动物分类学报, 2006, 31(3):520-524.
LI Q F, ZHAO X B, LIU H L, et al. A review of the research on taxonomic status in yak (Poephagus)[J]. Acta Zootaxonomica Sinica, 2006, 31(3):520-524. (in Chinese)
[20] 范晴, 谢芝勋, 谢志勤, 等. 牛病毒性腹泻病毒抗原捕获ELISA方法的建立[J]. 动物医学进展, 2015, 36(2):4-2.
FAN Q, XIE Z X, XIE Z Q, et al. Development of antigen capture ELISA of detection bovine viral diarrhea virus[J]. Progress in Veterinary Medicine, 2015, 36(2):4-2. (in Chinese)
[21] GAO J F, LIU M Y, MENG X R, et al. Seroprevalence of bovine viral diarrhea infection in Yaks (Bos grunniens) on the Qinghai-Tibetan Plateau of China[J]. Trop Anim Health Prod, 2013, 45(3):791-793.
[22] KALAYCIOGLU A T. Bovine viral diarrhoea virus (BVDV) diversity and vaccination. A review[J]. Vet Q, 2007, 29(2):60-67.
[23] PECORA A, MALACARI D A, PEREZ AGUIRREBURUALDE M S, et al. Development of an APC-targeted multivalent E2-based vaccine against bovine viral diarrhea virus types 1 and 2[J]. Vaccine, 2015, 33(39):5163-5171.
[24] OSTACHUK A. Bovine viral diarrhea virus structural protein E2 as a complement regulatory protein[J]. Arch Virol, 2016, 161(7):1769-1782.
[25] RIDPATH J F, LOVELL G, NEILL J D, et al. Change in predominance of Bovine viral diarrhea virus subgenotypes among samples submitted to a diagnostic laboratory over a 20-year time span[J]. J Vet Diagn Invest, 2011, 23(2):185-193.
[26] TOPLAK I, SANDVIK T, BARLIC-MAGANJA D, et al. Genetic typing of bovine viral diarrhoea virus:most Slovenian isolates are of genotypes 1d and 1f[J]. Vet Microbiol, 2004, 99(3-4):175-185.
[27] ABE Y, TAMURA T, TORⅡ S, et al. Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan[J]. J Vet Med Sci, 2016, 78(1):61-70.
[28] OEM J K, CHUNG J Y, ROH S, et al. Characterization and phylogenetic analysis of Bovine viral diarrhea virus in brain tissues from nonambulatory (downer) cattle in Korea[J]. J Vet Diagn Invest, 2010, 22(4):518-523.
[29] LANYON S R, REICHEL M P. Bovine viral diarrhoea virus (‘pestivirus’) in Australia:to control or not to control[J]. Aust Vet J, 2014, 92(8):277-282.
[30] 王炜. 牛主要呼吸道病毒病血清学调查、牛病毒性腹泻病毒分离株鉴定及疫苗研究[D]. 北京:中国农业科学院, 2014.
WANG W. Serosurvey of major bovine respiratory viruses and identification of BVDV isolates and vaccine development[D]. Beijing:Chinese Academy of Agricultural Sciences, 2014. (in Chinese)