黑龍江部分地區(qū)野生鳥類AIV和NDV感染狀況的初步研究.pdf_第1頁
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1、東北林業(yè)大學(xué)碩士學(xué)位論文黑龍江部分地區(qū)野生鳥類AIV和NDV感染狀況的初步研究姓名:張智明申請(qǐng)學(xué)位級(jí)別:碩士專業(yè):生理學(xué)指導(dǎo)教師:華育平20070601東北林業(yè)人學(xué)顧‘L學(xué)位論文AbstractAvianinfluenza(A_1)andNewcastledisease(ND),whichwerecontagiousandwidespreadavaindiseaseaffectingmostspeciesofbirds,aretwomo

2、stseriousinfectiousdiseaseofpoultryMostrecentlytheoutbreaksofNDVappearsnewcharacteristicThereportsofimmunizationfallingoccurredincreasinglyAvianinfluenzaisaseriousdiseaseinpoultrythatcancausesevereeconomiclossesinmanycou

3、ntries,also渤directlyinfecthumanso,thegovernmentsintheworldattachimportancetothisdiseaseWildbirdsareconsideredapotentialreservoirandmaythereforeplayaroleintheepidemiologyofeconomicallyimportantorzoonoticdiseaseStadingthep

4、revalenceofthistwodiseases,wemayfindouttheroleofwildbirdsinthetwovirusesspreadingFurthermore,itisveryimportantfurthetwodiseasesthatwecouldpreventandcontrolthemmoreeffectivelyOnehundredandeighty—threewildbirdswerecaptured

5、inHeiLongjiangprovincethentrachealandcloacalswabswc豫takenfromeachbirdusingcommercialswabsForthedetectionofAIVandNOVviruseswereisolatedbytwoorthreepassagesusingembryonated,specific—pathogenfree(SPF)eggsPositivesamplesWCIe

6、furthertestedbyhaemagglutinationinhibition(HI)andRTPCRResultsshowedtheoverallseparationrateofAIVseropositivesampleswas115%whichwere21Andcomprising16samplesthatweIcseropositiveforNDV(prevalence=78%、Nexttwelvespecificprime

7、rsfortheeightgenesegmentsofAIVweredesignedonthebasisofpublishedAIVnuclcotidesequencedatawiththeaidofprimerpremiersoftwareAndtheneightgenesegmentsofAIVfromwaterfowlWasamplifiedbyRToPCRwhichHA、PA、PBl、PB2we咒amplifiedbydivid

8、ingtwopartsThePCRproductswerepurified,directlyclonedintothevectorpMDl8TtransformedintoEcoilandobtainedthepositiveplasmidsforsequenceconfirmationFinallyweeditednucleotidesequences,analysis,predictionofaminoacidsequencesan

9、dalignmentswereconductedusingthebiologicalsoftwareAminoacidanalysisandphylogenetictreewereconstructedwithH6N2SequenceresultsshowedthattheeightgenesegmentsofAIVallcontainingcompleteopenreadingframeand5’3untranslatedregion

10、sBLASTcompareindicatedthisstrainoriginedfromEurasiawhichmaybehomologywithHokkaidostrainsNucleotidesequenceofHAgenepointedthatithasthesamecleavagesitewithlowpathogenicAIVHAgenewhichaminoacidis340PQIETRlG346BasedonH6N2subt

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