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

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：張智明申請學(xué)位級別：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