Abstract
This study was performed to investigate the prevalence and genetic variation of hepatitis E virus (HEV) in Tibetan pigs and to determine its ability to infect mice. A total of 38 out of 229 (16.59%; 95% CI = 12.00%–22.10%) fecal samples from Tibetan pigs from the Qinghai-Tibetan Plateau in 2018 were positive for HEV RNA, which was detected by RT-nPCR. Significantly different detection rates were observed between samples from diarrheic and clinically healthy animals (OR = 9.56; 95% CI, 2.84–32.14; p < 0.001), suggesting a potential association between HEV infection and diarrhea in Tibetan pigs. Phylogenetic analysis showed that the HEV isolates were clustered into subtypes 4a (31 samples), 4b (1), 4d (2), and 4j (4). HEV-4a was the predominant subtype, indicating that it might be circulating in Tibetan pigs. Nine complete HEV genome sequences obtained from Tibetan pigs were found by phylogenetic analysis to be closely related to those of genotype 4 HEV isolates from humans. Two recombinant events were identified in both HEV-4a strains; a novel recombination breakpoint was first identified at the 3’ end of the ORF2 region in the SWU/L9/2018 strain, and a common recombination region was found at the junction of the ORF1 and ORF2 regions in the SWU/31-12/2018 strain. Furthermore, HEV-4a could be detected in all BALB/c mice that were experimentally infected by gavage and contact exposure. The information presented here about the prevalence and genotype diversity of HEV from Tibetan pigs provides important insights into the epidemic features of HEV on the Qinghai-Tibetan Plateau.
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References
Meng XJ (2013) Zoonotic and foodborne transmission of hepatitis E virus. Semin Liver Dis 33(1):41–49. https://doi.org/10.1055/s-0033-1338113
Nan Y, Wu C, Zhao Q, Zhou EM (2017) Zoonotic hepatitis E virus: an ignored risk for public health. Front Microbiol 8:2396. https://doi.org/10.3389/fmicb.2017.02396
Meng XJ (2010) Hepatitis E virus: animal reservoirs and zoonotic risk. Vet Microbiol 140(3–4):256–265. https://doi.org/10.1016/j.vetmic.2009.03.017
Al-Sadeq DW, Majdalawieh AF, Mesleh AG, Abdalla OM, Nasrallah GK (2018) Laboratory challenges in the diagnosis of hepatitis E virus. J Med Microbiol 67(4):466–480. https://doi.org/10.1099/jmm.0.000706
Nan Y, Zhang YJ (2016) Molecular biology and infection of hepatitis E virus. Front Microbiol 7:1419. https://doi.org/10.3389/fmicb.2016.01419
Smith DB, Simmonds P, Izopet J, Oliveira-Filho EF, Ulrich RG, Johne R, Koenig M, Jameel S, Harrison TJ, Meng XJ, Okamoto H, Van der Poel WH, Purdy MA (2016) Proposed reference sequences for hepatitis E virus subtypes. J Gen Virol 97(3):537–542. https://doi.org/10.1099/jgv.0.000393
Woo PC, Lau SK, Teng JL, Cao KY, Wernery U, Schountz T, Chiu TH, Tsang AK, Wong PC, Wong EY (2013) Yuen KY (2016) New hepatitis E virus genotype in bactrian camels, Xinjiang, China. Emerg Infect Dis 22(12):2219–2221. https://doi.org/10.3201/eid2212.160979
Lee GH, Tan BH, Teo EC, Lim SG, Dan YY, Wee A, Aw PP, Zhu Y, Hibberd ML, Tan CK, Purdy MA, Teo CG (2016) Chronic infection with Camelid hepatitis E virus in a liver transplant recipient who regularly consumes camel meat and milk. Gastroenterology 150(2):355–357 e353. https://doi.org/10.1053/j.gastro.2015.10.048
Chan MCW, Kwok K, Hung TN, Chan PKS (2017) Molecular epidemiology and strain comparison between hepatitis E viruses in human sera and pig livers during 2014 to 2016 in Hong Kong. J Clin Microbiol 55(5):1408–1415. https://doi.org/10.1128/JCM.02020-16
Liu P, Li L, Wang L, Bu Q, Fu H, Han J, Zhu Y, Lu F, Zhuang H (2012) Phylogenetic analysis of 626 hepatitis E virus (HEV) isolates from humans and animals in China (1986–2011) showing genotype diversity and zoonotic transmission. Infect Genet Evol 12(2):428–434. https://doi.org/10.1016/j.meegid.2012.01.017
Li H, Wu J, Sheng Y, Lu Q, Liu B, Chen Y, Sun Y, Zhou EM, Zhao Q (2019) Prevalence of hepatitis E virus (HEV) infection in various pig farms from Shaanxi Province, China: first detection of HEV RNA in pig semen. Transbound Emerg Dis 66(1):72–82. https://doi.org/10.1111/tbed.12966
Zhou JH, Wang YN, Li XR, Liu YS, Pan Q, Lan X (2018) Prevalence and genetic features of hepatitis E virus in swine, in Gansu, China. Acta Virol 62(2):196–201. https://doi.org/10.4149/av_2018_212
Li K, Zhang L, Zhang H, Lei Z, Luo H, Mehmood K, Shahzad M, Lan Y, Wang M, Li J (2017) Epidemiological investigation and risk factors of Echinococcus granulosus in yaks (Bos grunniens), Tibetan pigs and Tibetans on Qinghai Tibetan plateau. Acta Trop 173:147–152. https://doi.org/10.1016/j.actatropica.2017.06.019
Wu Q, An J, She R, Shi R, Hao W, Soomro M, Yuan X, Yang J, Wang J (2017) Detection of genotype 4 swine hepatitis E virus in systemic tissues in cross-species infected rabbits. PLoS One 12(1):e0171277. https://doi.org/10.1371/journal.pone.0171277
Gongga, Wang Y, Yixicuomu, Qiongda, Suolangsizhu (2018) Epidemiological investigation of hepatitis E virus infection in Tibetan swine population. Sheng Wu Gong Cheng Xue Bao 34(9):1453–1459. https://doi.org/10.13345/j.cjb.180189
Zhang L, Huang S, Li K, Rehman MU, Jiang X, Tong X, Zhang H, Iqbal MK, Mehmood K, Liu S, Shen Y, Li J (2018) Molecular detection of indigenous hepatitis E virus (HEV) from Tibetan Pigs in Tibet, China. Food Environ Virol 10(4):373–377. https://doi.org/10.1007/s12560-018-9352-6
Zhang L, Li K, Huang S, Liu D, Rehman MU, Lan Y, Zhang H, Wang L, Hao Y, Iqbal MK, Mehmood K, Chamba Y, Li J (2017) Seroprevalence and risk factors associated with hepatitis E virus infections among people and pigs in Tibet, China. Acta Trop 172:102–106. https://doi.org/10.1016/j.actatropica.2017.04.033
Li F, Meng JH, Dong C, Dai X, Yang YG, Zhou ZX (2009) Design and application of a set of universal PCR primers for genotyping of hepatitis E virus. Bing Du Xue Bao 25(1):9–16 (in Chinese)
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33(7):1870–1874. https://doi.org/10.1093/molbev/msw054
Martin DP, Murrell B, Golden M, Khoosal A, Muhire B (2015) RDP4: detection and analysis of recombination patterns in virus genomes. Virus Evol 1(1):vev003. https://doi.org/10.1093/ve/vev003
Sun Y, Lu Q, Liu B, Sheng Y, Du T, Hiscox JA, Zhou EM, Zhao Q (2018) Cross-species infection of mice by rabbit hepatitis E virus. Vet Microbiol 225:48–52. https://doi.org/10.1016/j.vetmic.2018.09.015
Wang XJ, Zhao Q, Jiang FL, Liu BY, Zhao JN, Dang L, Sun YN, Mu Y, Xiao SQ, Wang CB, Hsu WH, Liu L, Widen F, Zhou EM (2014) Genetic characterization and serological prevalence of swine hepatitis E virus in Shandong province, China. Vet Microbiol 172(3–4):415–424. https://doi.org/10.1016/j.vetmic.2014.06.006
Cao M, Liu F, Jiang S, Shao X, Lan K, Li X, Zhang Z, Wang W, Zhao W, Huang R (2002) Seroepidemiologic survey of hepatitis G virus in selected population of Shanxi, Qinghai and Xinjiang Provinces (region) of China. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 16(4):345–347
Qing Y, Juanjuan Q, Min Y, Liping Z, Wenhua X (2009) Prevalence of hepatitis E virus antibodies in human and livestock in Qinghai region. Chin J Vet Med 71:71–72 (in Chinese)
Toole MJ, Claridge F, Anderson DA, Zhuang H, Morgan C, Otto B, Stewart T (2006) Hepatitis E virus infection as a marker for contaminated community drinking water sources in Tibetan villages. Am J Trop Med Hyg 74(2):250–254
Gao X, Zhang L, Jiang X, Mehmood K, Wang L, Tong X, Wang M, Zhang H, Li J (2019) Porcine epidemic diarrhea: an emerging disease in Tibetan pigs in Tibet, China. Trop Anim Health Prod 51(2):491–494. https://doi.org/10.1007/s11250-018-1707-3
Wang M, Wang Y, Baloch AR, Pan Y, Tian L, Xu F, Shivaramu S, Chen S, Zeng Q (2018) Detection and genetic characterization of porcine deltacoronavirus in Tibetan pigs surrounding the Qinghai-Tibet Plateau of China. Transbound Emerg Dis 65(2):363–369. https://doi.org/10.1111/tbed.12819
Monini M, Di Bartolo I, Ianiro G, Angeloni G, Magistrali CF, Ostanello F, Ruggeri FM (2015) Detection and molecular characterization of zoonotic viruses in swine fecal samples in Italian pig herds. Arch Virol 160(10):2547–2556. https://doi.org/10.1007/s00705-015-2538-4
Liu B, Chen Y, Sun Y, Nan Y, Li H, Du T, Hiscox JA, Zhao Q, Zhou EM (2019) Experimental infection of rabbit with swine-derived hepatitis E virus genotype 4. Vet Microbiol 229:168–175. https://doi.org/10.1016/j.vetmic.2019.01.001
Lu YH, Qian HZ, Hu AQ, Ren H, Qin X, Jiang QW, Zheng YJ (2014) Duration of viraemia in Chinese acute sporadic hepatitis E. Eur J Clin Microbiol Infect Dis 33(5):755–759. https://doi.org/10.1007/s10096-013-2007-5
Kelly AG, Netzler NE, White PA (2016) Ancient recombination events and the origins of hepatitis E virus. BMC Evol Biol 16(1):210. https://doi.org/10.1186/s12862-016-0785-y
Wang H, Zhang W, Ni B, Shen H, Song Y, Wang X, Shao S, Hua X, Cui L (2010) Recombination analysis reveals a double recombination event in hepatitis E virus. Virol J 7:129. https://doi.org/10.1186/1743-422X-7-129
Chen X, Zhang Q, He C, Zhang L, Li J, Zhang W, Cao W, Lv YG, Liu Z, Zhang JX, Shao ZJ (2012) Recombination and natural selection in hepatitis E virus genotypes. J Med Virol 84(9):1396–1407. https://doi.org/10.1002/jmv.23237
van Cuyck H, Fan J, Robertson DL, Roques P (2005) Evidence of recombination between divergent hepatitis E viruses. J Virol 79(14):9306–9314. https://doi.org/10.1128/JVI.79.14.9306-9314.2005
Debing Y, Mishra N, Verbeken E, Ramaekers K, Dallmeier K, Neyts J (2016) A rat model for hepatitis E virus. Dis Model Mech 9(10):1203–1210. https://doi.org/10.1242/dmm.024406
Cossaboom CM, Cordoba L, Sanford BJ, Pineyro P, Kenney SP, Dryman BA, Wang Y, Meng XJ (2012) Cross-species infection of pigs with a novel rabbit, but not rat, strain of hepatitis E virus isolated in the United States. J Gen Virol 93(Pt 8):1687–1695. https://doi.org/10.1099/vir.0.041509-0
Yu W, Yang C, Hao X, Ma T, Huang F (2018) Successful infection of BALB/c mice by a swine hepatitis E virus clone constructed with reverse genetics. BMC Infect Dis 18(1):687. https://doi.org/10.1186/s12879-018-3544-4
Acknowledgements
Special thanks to Dr. Yuchen Nan of Northwest A&F University, China who kindly donated the purified sORF2-C protein and mouse monoclonal antibody against HEV. This study was supported by funding by the National Natural Science Foundation of China (Grant no. 31772766), the 13th Five-Year Plan National Key Research and Development Program of China (Grant no. 2016YFD0500705), the Innovative Research Team Program of the Department of Education of Sichuan Province (Grant no. 13TD0057), and the Fundamental Research Funds for the Central Universities of Southwest Minzu University (Grant no. 2017NZYQN01).
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Zhou, K., Zhou, Q., Yang, D. et al. Hepatitis E virus in Tibetan pigs on the Qinghai-Tibetan Plateau: detection, molecular characterization and infection of BALB/c mice. Arch Virol 164, 2943–2951 (2019). https://doi.org/10.1007/s00705-019-04410-x
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DOI: https://doi.org/10.1007/s00705-019-04410-x