research article first report of hepatitis e virus infection in sika...

Research ArticleFirst Report of Hepatitis E Virus Infection inSika Deer in China

Xiao-Xuan Zhang12 Si-Yuan Qin12 Yuan Zhang1 Qing-Feng Meng3

Jing Jiang1 Gui-Lian Yang1 Quan Zhao1 and Xing-Quan Zhu24

1College of Animal Science and Technology Jilin Agricultural University Changchun Jilin 130118 China2State Key Laboratory of Veterinary Etiological Biology Lanzhou Veterinary Research InstituteChinese Academy of Agricultural Sciences Lanzhou Gansu 730046 China3Jilin Entry-Exit Inspection and Quarantine Bureau Changchun Jilin 130000 China4Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Disease Yangzhou Jiangsu 225009 China

Correspondence should be addressed to Quan Zhao zhaoquan0825163com and Xing-Quan Zhu xingquanzhu1hotmailcom

Received 20 January 2015 Revised 25 March 2015 Accepted 25 March 2015

Academic Editor Mohamed T Shata

Copyright copy 2015 Xiao-Xuan Zhang et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Hepatitis E virus (HEV) a single stranded RNA nonenveloped virus belongs to the genusHepevirus in the family ofHepeviridaeIn this study 46 (543) out of the 847 serum samples from sika deer (Cervus nippon) were detected as seropositive with hepatitisE virus (HEV) by enzyme linked immunosorbent assay (ELISA) These samples were collected from Inner Mongolia and Jilinand Heilongjiang provinces in China between October 2012 and October 2013 Seroprevalence of HEV infection in male andfemale deer was 482 and 652 respectively HEV seroprevalence in sika deer from different geographical locations varied from313 to 673 There was no significant difference in HEV seroprevalence between sika deer collected in autumn (565) andwinter (485)This is the first report of HEV seroprevalence in sika deer in China which will provide foundation information forestimating the effectiveness of future measures to control HEV infection in sika deer in China and assessing the potential risk ofhumans infected with HEV after consumption of undercooked or raw meat from infected sika deer

1 Introduction

Viral hepatitis is a global health problem [1] especially hep-atitis E HEV is the causative agent of hepatitis E [2] HepatitisE virus (HEV) a single stranded RNA nonenveloped virusbelongs to the genus Hepevirus in the family of Hepeviridae[2ndash6] Previous studies introduced four routes to transmis-sion of HEV They are fecal-oral transmission foodbornetransmission transfusion and vertical transmission [7]HEVinfection can lead to protracted coagulopathy and cholestatisand is also associated with death especially among pregnantwomen [7ndash10]

Hepatitis E was first recognized in India in 1978 [5]Avian HEV and mammalian HEV are major species ofhepatitis E virus [5] More than four genotypes (genotypes1 2 3 and 4) have been identified in HEV [6 11 12]for example genotypes 1 and 2 of HEV were commonly

prevalent in developing countries and genotype 2 appearsto be exclusively anthroponotic while genotype 1 infectsmainly humans but has also been detected in pigs [13 14]Genotype 3was distributed around theworld and could causesevere HEV infection in humans and animals Most of HEVgenotype 4 infections were in Asian countries and they havebeen found in humans and pigs [11 15ndash18] Recently genotype5 has been proposed as the designation for a variant of HEVisolated fromwild boars [12] Haqshenas et al [19] andHuanget al [20] identified a novel HEV strain which was isolatedfrom chicken but Avian HEV could not infect nonhumanprimates [20] Humans can be infected by six genotypes ofmammalian HEV namely types 1 to 4 and two additionalgenotypes from wild boar [5]

In view of such serious situation a large number ofinvestigations about HEV have been carried out worldwide[11ndash27] In China infection of HEV has also been reported

Hindawi Publishing CorporationBioMed Research InternationalVolume 2015 Article ID 502846 5 pageshttpdxdoiorg1011552015502846

2 BioMed Research International

Inner Mongolia

Heilongjiang

Jilin

Zhang et al first report of hepatitis E virus infection in sika deer in China

Figure 1 Map showing Heilongjiang province Jilin province and Inner Mongolia Autonomous Region stranded out in white color innortheastern China where sika deer serum samples were collected for detection of hepatitis E virus antibodies

in wild rats rabbits pigs sheep parrots chickens dogs catsand humans [3 29ndash35] but no information was availableaboutHEV infection in sika deerTheobjectives of this surveywere to investigate the distribution and seroprevalence ofHEV infection in sika deer from 4 cities Northern China forthe first time provide the ldquobaselinerdquo data for the control ofHEV infection and assess the potential risk that people areinfected by HEV

2 Materials and Methods

21 Ethics Statement This study was approved by the AnimalEthics Committee of Lanzhou Veterinary Research InstituteChinese Academy of Agricultural Sciences (Approval num-ber LVRIAEC2012-008) The sika deer from which bloodwas collected were handled in accordance with good animalpractices required by the Animal Ethics Procedures andGuidelines of the Peoplersquos Republic of China

22 The Study Site The samples in this study were collectedfrom4 citiesNorthernChina (Figure 1) Changchun (43∘051015840sim45∘151015840N 124∘181015840 sim127∘051015840E) is the capital of Jilin province andit is one of the central cities in Northeast China and belongsto a semiwet monsoon type zone Jilin City (42∘311015840sim44∘401015840N125∘401015840 sim127∘561015840E) is located in Jilin province and the climateof Jilin City is northerly continental monsoon type Harbin(44∘041015840sim46∘401015840N 125∘421015840sim130∘101015840E) is one of the central citiesin Northeast China and the capital of Heilongjiang provinceHarbin belongs to a temperate continentalmonsoon type climateChifeng (41∘1710158401010158401015840sim45∘2410158401510158401015840N 116∘2110158400710158401015840sim120∘5810158405210158401015840E) islocated in the upper reaches of the Xiliao River Chifeng has

a local steppe climate Sika deer are mostly originated inChangchun Chifeng and Harbin in Northern China

23 Serum Samples All the sika deer blood samples (208from Harbin city 316 from Changchun city 163 from JilinCity and 160 from Chifeng city) were randomly collectedfrom the 4 cities Northern China between October 2012 andOctober 2013 All the sika deer were bred in the semifreerange system Samples were transported to the laboratoryin the College of Animal Science and Technology JilinAgriculture University Jilin province China They were keptat 37∘C for 2 h and then at 4∘C for 1 h and then centrifugationwas carried out at 1000timesg for 10 min and the serumwas separated and kept at minus20∘C until further analysisInformation of sika deer about seasons and geographic originof sampling gender and age was recorded

24 Serological Examination Circulating antibodies (CAb)against HEV were tested by the species-independent double-antigen sandwich (das) ELISA using a commercially availablekit (MPBiomedicals Asia Pacific Pte Ltd Singapore) accord-ing to themanufacturerrsquos instructions [22] Positive negativeand blank controlswere supplied in the kit and set in each test

25 Statistical Analysis The differences of seroprevalence ofHEV infection in sika deer from different locations genderand seasons were analyzed statistically using SAS software(version 91 SAS Institute Inc Cary NC) [36] Resultswere considered statistically significant if 119875 lt 005 Oddsratios (OR) and their 95 confidence interval (95 CI) wereprovided in this study

BioMed Research International 3

Table 1 Seroprevalence of hepatitis E virus (HEV) infection in sika deer (Cervus nippon) in different region gender and season by enzymelinked immunosorbent assay (ELISA)

Variable Category Number ofexamined deer

Number ofpositive deer Prevalence () (95 CI) 119875 value OR (95 CI)

Gender Male 540 26 482 (301ndash662) 029 ReferenceFemale 307 20 652 (375ndash928) 138 (076ndash251)

Region

Harbin 208 14 673 (333ndash1014)

036

ReferenceChangchun 316 20 633 (364ndash901) 094 (046ndash190)Jilin City 163 7 429 (118ndash741) 062 (025ndash158)Chifeng 160 5 313 (043ndash582) 045 (016ndash127)

Season Autumn 620 35 565 (383ndash746) 065 ReferenceWinter 227 11 485 (205ndash764) 085 (043ndash171)

Total 847 46 543 (391ndash696)

Table 2 Prevalence of hepatitis E virus (HEV) infection in deer around the world

Country Species Testa Number of tested deer Positive () Yearb ReferencePoland Red deer (Cervus elaphus) ELISA 118 0 2012-2013 [21]Poland Roe deer (Capreolus capreolus) ELISA 38 0 2012-2013 [21]Poland Fallow deer (Dama dama) ELISA 5 0 2012-2013 [21]Poland Sika deer (Cervus nippon) ELISA 4 0 2012-2013 [21]Netherlands Red deer ELISA 38 5 2005ndash2008 [22]Netherlands Red deer RT-PCR 39 15 2005ndash2008 [22]Japan Deer ELISA 117 2 2003-2004 [23]USA Wild sika deer ELISA 174 0 UN [24]Japan Wild sika deer ELISA 976 26 UN [25]Japan Wild sika deer RT-PCR 247 0 UN [26]Japan Yezo deer ELISA 520 348 UN [26]Japan Yezo deer WB 520 28 UN [26]Spain Red deer ELISA 968 104 2000ndash2009 [27]Spain Red deer RT-PCR 968 136 2000ndash2009 [27]Hungary Roe deer RT-PCR 32 344 2001ndash2006 [28]aELISA enzyme-linked immunosorbent assay RT-PCR reverse transcription polymerase chain reaction WB Western blotbUN unknown

3 Results

A total of 46 (543 95 confidence interval (CI) 391ndash696)out of 847 sika deer were detected as seropositive of HEVby ELISA (Table 1) Seroprevalence of HEV in male deerwas 482 (95 CI 301ndash662) which was lower than that infemale deer (652 95 CI 375ndash928) but the difference wasnot statistically significant (119875 gt 005) HEV seroprevalencein sika deer from different geographical locations variedfrom 313 (95 CI 043ndash582) to 673 (95 CI 333ndash1014)but the difference was not statistically significant amongthe different regions (119875 gt 005) Sika deer collected inwinter (485 95 CI 205ndash764 119875 gt 005) had a slightlylower HEV seroprevalence compared to sika deer collectedin autumn (565 95 CI 383ndash746) and the difference wasnot significant statistically

4 Discussion

There are many reports showing that HEV could infect deerall over the world (Table 2) In the present study we surveyed

the HEV seroprevalence in sika deer at 4 cities in NorthernChina The overall HEV seroprevalence in sika deer was543 whichwas lower than that in red deer (Cervus elaphus)in Spain (104) [27] and Yezo deer in Japan (348) [26] byELISA and in red deer in Spain (136) [27] and Netherlands(15) [22] and roe deer in Hungary (344) [28] by RT-PCRbut higher than that in red deer in Netherlands (5) [22]and wild sika deer in Japan (26) [25] and USA (0) [7] byELISA and in wild sika deer in Japan by RT-PCR (0) [25]and Western blot (28) [26] (Table 2) It is well known thatdue to high discordance between assays in different detectionmethods the actual discrepancy is difficult to explain in theprevalence of HEV among different studies The differencesmay also be due to the investigated areas that challenged thesurvival of HEV the differences in sanitation animal-welfarefor deer animal husbandry practices and geographical andecological factors for example rainfall

Previous studies [29 37] demonstrated the male has thehigher positive rates than the female but the present studyindicates the opposite although the difference in prevalence


between females and males is not statistically significant(Table 1) In different regions group HEV seroprevalence insika deer varied from 313 (5160) to 673 (14208) Sikadeer in Harbin has the highest HEV seroprevalence followedby Changchun the third was Jilin City and Chifeng was thelast but the difference was not statistically significant (119875 =036) (Table 1) Climate geography degree of environmentcontamination with HEV ecological conditions feeding con-ditions and animal welfares could be the reasons for thedifferences in HEV seroprevalence in sika deer in differentcities

In terms of the sampling seasons HEV seroprevalence insika deer collected in autumn was 565 which was higherthan that collected in winter (485) but the difference wasnot statistically significant (119875 = 065) (Table 1) This mayhappen because in autumn the local climate was suitablefor the survival of HEV and the resistance of sika deer wasimproved in winter in four cities Compared with autumnsika deer has a small range of activities and the relatively lowerexposure of HEV in winter

Sika deer are popular in China and they are famous formedicinal value Velvet antlers blood and meat are the mainproducts from sika deer Deer meat especially was widelyrecognized by most people with the improvement of livingstandards in Northern China A study has shown that HEVcould be transmitted from deer to humans [37] so sika deeris considered as the potential source for the spread of HEV tohumans but in this study we did not demonstrate that theelder have higher HEV seroprevalence than the younger [38]because the samples were collected in four cities in northernChina betweenOctober 2012 andOctober 2013 just includingautumn and winter samples and the animals were adult sikadeer so our results may have not reflected the relationshipbetween HEV seroprevalence and age Moreover the studyonly detected the antibodies against HEV in sika deer and nosequence information was obtained due to limited volume ofserum so it could not reflect the true infection rate Furtherstudies should be conducted to get sequence information toconfirm the real HEV infection rates and to determine theHEV genotypes

5 Conclusion

Thepresent investigation suggests the existence ofHEV infec-tion in sika deer Northern China The result may providefundamental information for estimating the effectiveness offuturemeasure to controlHEV infection in sika deer inChinaand assessing the potential risk of humans infected by HEV

Conflict of Interests

All the authors declare no conflict of interests

Authorsrsquo Contribution

Xiao-Xuan Zhang and Si-Yuan Qin contributed equally tothis work

Acknowledgments

This study was supported by the Key Scientific and Techno-logical Project of Jilin Province (Grant no 20140204068NY)and the Science Fund for Creative Research Groups of GansuProvince (Grant no 1210RJIA006)

References

[1] J KhanM Shafiq SMushtaq et al ldquoSeropositivity and coinfec-tion of hepatitis B andC among patients seeking hospital care inIslamabad Pakistanrdquo BioMed Research International vol 2014Article ID 516859 4 pages 2014

[2] S A RutjesM Bouwknegt JW van derGiessenAM deRodaHusman andC B EM Reusken ldquoSeroprevalence of hepatitis evirus in pigs fromdifferent farming systems in theNetherlandsrdquoJournal of Food Protection vol 77 no 4 pp 640ndash642 2014

[3] X-X ZhangN-Z ZhangD-H ZhouW-P Tian Y-T Xu andX-Q Zhu ldquoSeroprevalence of hepatitis E virus in four speciesof parrots in Chinardquo Pakistan Veterinary Journal vol 34 no 1pp 130ndash132 2014

[4] W F El-Tras A A Tayel and N N El-Kady ldquoSeroprevalenceof hepatitis E virus in humans and geographicallymatched foodanimals in Egyptrdquo Zoonoses and Public Health vol 60 no 3 pp244ndash251 2013

[5] M S Khuroo ldquoDiscovery of hepatitis E the epidemic non-A non-B hepatitis 30 years down the memory lanerdquo VirusResearch vol 161 no 1 pp 3ndash14 2011

[6] I KMushahwar ldquoHepatitis E virus molecular virology clinicalfeatures diagnosis transmission epidemiology and preven-tionrdquo Journal of Medical Virology vol 80 no 4 pp 646ndash6582008

[7] B Xu H B Yu W Hui et al ldquoClinical features and risk factorsof acute hepatitis E with severe jaundicerdquo World Journal ofGastroenterology vol 18 no 48 pp 7279ndash7284 2012

[8] TNChau S T Lai C Tse et al ldquoEpidemiology and clinical fea-tures of sporadic hepatitis E as compared with hepatitis ArdquoTheAmerican Journal of Gastroenterology vol 101 no 2 pp 292ndash296 2006

[9] C Renou V Gobert C Locher et al ldquoProspective study ofHepatitis E Virus infection among pregnant women in FrancerdquoVirology Journal vol 11 no 1 article 68 2014

[10] Y Song W Park B Park et al ldquoHepatitis E virus infectionsin humans and animalsrdquo Clinical and Experimental VaccineResearch vol 3 no 1 pp 29ndash36 2014

[11] G G Schlauder and I K Mushahwar ldquoGenetic heterogeneityof hepatitis E virusrdquo Journal of Medical Virology vol 65 no 2pp 282ndash292 2001

[12] M Takahashi T Nishizawa H Sato Y Sato S Nagashima andH Okamoto ldquoAnalysis of the full-length genome of a HepatitisE virus isolate obtained from a wild boar in Japan that isclassifiable into a novel genotyperdquo Journal of General Virologyvol 92 no 4 pp 902ndash908 2011

[13] J E Arends V Ghisetti W Irving et al ldquoHepatitis E an emerg-ing infection in high income countriesrdquo Journal of ClinicalVirology vol 59 no 2 pp 81ndash88 2014

[14] A Ahmed I A Ali H Ghazal J Fazili and S Nusrat ldquoMysteryof hepatitis e virus recent advances in its diagnosis andmanagementrdquo International Journal of Hepatology vol 2015Article ID 872431 6 pages 2015


[15] X J Meng P G Halbur M S Shapiro et al ldquoGenetic andexperimental evidence for cross-species infection by swinehepatitis E virusrdquo Journal of Virology vol 72 no 12 pp 9714ndash9721 1998

[16] L Lu C Li and C H Hagedorn ldquoPhylogenetic analysis ofglobal hepatitis E virus sequences genetic diversity subtypesand zoonosisrdquo Reviews in Medical Virology vol 16 no 1 pp 5ndash36 2006

[17] F R Lorenzo B Tsatsralt-Od S Ganbat M Takahashi andH Okamoto ldquoAnalysis of the full-length genome of hepatitis Evirus isolates obtained from farm pigs in Mongoliardquo Journal ofMedical Virology vol 79 no 8 pp 1128ndash1137 2007

[18] H Okamoto ldquoGenetic variability and evolution of hepatitis Evirusrdquo Virus Research vol 127 no 2 pp 216ndash228 2007

[19] G Haqshenas H L Shivaprasad P R Woolcock D H Readand X J Meng ldquoGenetic identification and characterization ofa novel virus related to human hepatitis E virus from chickenswith hepatitis-splenomegaly syndrome in the United StatesrdquoThe Journal of General Virology vol 82 no 10 pp 2449ndash24622001

[20] F F Huang Z F Sun S U Emerson et al ldquoDetermination andanalysis of the complete genomic sequence of avian hepatitis Evirus (avian HEV) and attempts to infect rhesus monkeys withavian HEVrdquo The Journal of General Virology vol 85 no 6 pp1609ndash1618 2004

[21] M Larska M K Krzysiak A Jabłonski J Kesik M Bednarskiand J Rola ldquoHepatitis E virus antibody prevalence in wildlife inPolandrdquo Zoonoses and Public Health vol 62 no 2 pp 105ndash1102015

[22] S A Rutjes F Lodder-Verschoor W J Lodder et al ldquoSero-prevalence and molecular detection of hepatitis E virus in wildboar and red deer in The Netherlandsrdquo Journal of VirologicalMethods vol 168 no 1-2 pp 197ndash206 2010

[23] H Sonoda M Abe T Sugimoto et al ldquoPrevalence of hepatitisE virus (HEV) infection in wild boars and deer and geneticidentification of a genotype 3 HEV from a boar in JapanrdquoJournal of Clinical Microbiology vol 42 no 11 pp 5371ndash53742004

[24] C Yu C Zimmerman R Stone et al ldquoUsing improved tech-nology for filter paper-based blood collection to survey wildSika deer for antibodies to hepatitis E virusrdquo Journal of Viro-logical Methods vol 142 no 1-2 pp 143ndash150 2007

[25] Y Matsuura M Suzuki K Yoshimatsu et al ldquoPrevalence ofantibody to hepatitis e virus among wild sika deer Cervusnippon in Japanrdquo Archives of Virology vol 152 no 7 pp 1375ndash1381 2007

[26] D Tomiyama E Inoue Y Osawa and K Okazaki ldquoSerologicalevidence of infection with hepatitis e virus among wild Yezo-deer Cervus nippon yesoensis in Hokkaido Japanrdquo Journal ofViral Hepatitis vol 16 no 7 pp 524ndash528 2009

[27] M Boadella M Casas M Martın et al ldquoIncreasing Contactwith hepatitis E virus in red deer Spainrdquo Emerging InfectiousDiseases vol 16 no 12 pp 1994ndash1996 2010

[28] G Reuter D Fodor P Forgach A Katai and G SzucsldquoCharacterization and zoonotic potential of endemic hepatitisE virus (HEV) strains in humans and animals in HungaryrdquoJournal of Clinical Virology vol 44 no 4 pp 277ndash281 2009

[29] W Li D Guan J Su et al ldquoHigh prevalence of rat hepatitis Evirus in wild rats in Chinardquo Veterinary Microbiology vol 165no 3-4 pp 275ndash280 2013

[30] S Wang C Dong X Dai et al ldquoHepatitis E virus isolated fromrabbits is genetically heterogeneous but with very similar

antigenicity to human HEVrdquo Journal of Medical Virology vol85 no 4 pp 627ndash635 2013

[31] X J Wang Q Zhao F L Jiang et al ldquoGenetic characterizationand serological prevalence of swine hepatitis E virus in Shan-dong province Chinardquo Veterinary Microbiology vol 172 no 3-4 pp 415ndash424 2014

[32] J-Y Wu Q Kang W-S Bai and Z-H Bai ldquoSeroepidemiologi-cal survey of sheep hepatitis E virus infection in Aksu region ofXinjiang Autonomousrdquo Chinese Journal of Virology vol 26 no3 pp 234ndash237 2010 (Chinese)

[33] Q Zhao E M Zhou S W Dong et al ldquoAnalysis of avianhepatitis E virus from chickens Chinardquo Emerging InfectiousDiseases vol 16 no 9 pp 1469ndash1472 2010

[34] H Liang J Chen J Xie et al ldquoHepatitis E virus serosurveyamong pet dogs and cats in several developed cities in ChinardquoPLoS ONE vol 9 no 6 Article ID e98068 2014

[35] X Dai C Dong Z Zhou et al ldquoHepatitis E virus genotype 4Nanjing China 2001ndash2011rdquo Emerging Infectious Diseases vol19 no 9 pp 1528ndash1530 2013

[36] L Zhang H Liu B Xu et al ldquoRural residents in China are atincreased risk of exposure to tick-borne pathogens anaplasmaphagocytophilum and ehrlichia chaffeensisrdquo BioMed ResearchInternational vol 2014 Article ID 313867 11 pages 2014

[37] S Tei N Kitajima S Ohara et al ldquoConsumption of uncookeddeer meat as a risk factor for hepatitis E virus infection anage- and sex-matched case-control studyrdquo Journal of MedicalVirology vol 74 no 1 pp 67ndash70 2004

[38] C G Teo ldquoMuch meat much malady changing perceptionsof the epidemiology of hepatitis Erdquo Clinical Microbiology andInfection vol 16 no 1 pp 24ndash32 2010

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


Inner Mongolia

Heilongjiang

Jilin

Zhang et al first report of hepatitis E virus infection in sika deer in China

Figure 1 Map showing Heilongjiang province Jilin province and Inner Mongolia Autonomous Region stranded out in white color innortheastern China where sika deer serum samples were collected for detection of hepatitis E virus antibodies

in wild rats rabbits pigs sheep parrots chickens dogs catsand humans [3 29ndash35] but no information was availableaboutHEV infection in sika deerTheobjectives of this surveywere to investigate the distribution and seroprevalence ofHEV infection in sika deer from 4 cities Northern China forthe first time provide the ldquobaselinerdquo data for the control ofHEV infection and assess the potential risk that people areinfected by HEV

2 Materials and Methods

21 Ethics Statement This study was approved by the AnimalEthics Committee of Lanzhou Veterinary Research InstituteChinese Academy of Agricultural Sciences (Approval num-ber LVRIAEC2012-008) The sika deer from which bloodwas collected were handled in accordance with good animalpractices required by the Animal Ethics Procedures andGuidelines of the Peoplersquos Republic of China

22 The Study Site The samples in this study were collectedfrom4 citiesNorthernChina (Figure 1) Changchun (43∘051015840sim45∘151015840N 124∘181015840 sim127∘051015840E) is the capital of Jilin province andit is one of the central cities in Northeast China and belongsto a semiwet monsoon type zone Jilin City (42∘311015840sim44∘401015840N125∘401015840 sim127∘561015840E) is located in Jilin province and the climateof Jilin City is northerly continental monsoon type Harbin(44∘041015840sim46∘401015840N 125∘421015840sim130∘101015840E) is one of the central citiesin Northeast China and the capital of Heilongjiang provinceHarbin belongs to a temperate continentalmonsoon type climateChifeng (41∘1710158401010158401015840sim45∘2410158401510158401015840N 116∘2110158400710158401015840sim120∘5810158405210158401015840E) islocated in the upper reaches of the Xiliao River Chifeng has

a local steppe climate Sika deer are mostly originated inChangchun Chifeng and Harbin in Northern China

23 Serum Samples All the sika deer blood samples (208from Harbin city 316 from Changchun city 163 from JilinCity and 160 from Chifeng city) were randomly collectedfrom the 4 cities Northern China between October 2012 andOctober 2013 All the sika deer were bred in the semifreerange system Samples were transported to the laboratoryin the College of Animal Science and Technology JilinAgriculture University Jilin province China They were keptat 37∘C for 2 h and then at 4∘C for 1 h and then centrifugationwas carried out at 1000timesg for 10 min and the serumwas separated and kept at minus20∘C until further analysisInformation of sika deer about seasons and geographic originof sampling gender and age was recorded

24 Serological Examination Circulating antibodies (CAb)against HEV were tested by the species-independent double-antigen sandwich (das) ELISA using a commercially availablekit (MPBiomedicals Asia Pacific Pte Ltd Singapore) accord-ing to themanufacturerrsquos instructions [22] Positive negativeand blank controlswere supplied in the kit and set in each test

25 Statistical Analysis The differences of seroprevalence ofHEV infection in sika deer from different locations genderand seasons were analyzed statistically using SAS software(version 91 SAS Institute Inc Cary NC) [36] Resultswere considered statistically significant if 119875 lt 005 Oddsratios (OR) and their 95 confidence interval (95 CI) wereprovided in this study






Region

Harbin 208 14 673 (333ndash1014)

036



Total 847 46 543 (391ndash696)



3 Results


4 Discussion








5 Conclusion






Acknowledgments


References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















Region

Harbin 208 14 673 (333ndash1014)

036



Total 847 46 543 (391ndash696)



3 Results


4 Discussion








5 Conclusion






Acknowledgments


References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















5 Conclusion






Acknowledgments


References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















research article first report of hepatitis e virus infection in sika...

Documents