利用秀麗隱桿線蟲模式探討豬大腸桿菌的致病性

兩株具有多個毒素基因的臨床分離株對秀麗隱桿線蟲不具顯著致病性，代表這些毒素基因可能具有交互作用或基因表現的程度較低而影響大腸桿菌在秀麗隱桿線蟲的致病表現。

資料載入處理中... 跳到主要內容 臺灣博碩士論文加值系統 ::: 網站導覽| 首頁| 關於本站| 聯絡我們| 國圖首頁| 常見問題| 操作說明 English |FB專頁 |Mobile 免費會員 登入| 註冊 功能切換導覽列 (159.65.11.210)您好！臺灣時間：2022/08/1312:00 字體大小：       ::: 詳目顯示 recordfocus 第1筆/ 共1筆  /1頁 論文基本資料 摘要 外文摘要 目次 參考文獻 電子全文 紙本論文 QRCode 本論文永久網址: 複製永久網址Twitter研究生:洪嘉駿研究生(外文):Chia-ChunHung論文名稱:利用秀麗隱桿線蟲模式探討豬大腸桿菌的致病性論文名稱(外文):UsingtheCaenorhabditiselegansModeltoInvestigatethePathogenicityofEscherichiacoliinSwine指導教授:陳德勛指導教授(外文):Ter-HsinChen口試委員:陳志銘、徐維莉口試委員(外文):Chih-MingChen、Wei-LiHsu口試日期:2019-06-10學位類別:碩士校院名稱:國立中興大學系所名稱:獸醫病理生物學研究所學門:獸醫學門學類:獸醫學類論文種類:學術論文論文出版年:2019畢業學年度:107語文別:中文論文頁數:63中文關鍵詞:豬大腸桿菌、大腸桿菌毒素基因、秀麗隱桿線蟲外文關鍵詞:Escherichiacoliofswine、virulencefactorsofE.coli、Caenorhabditiselegans相關次數: 被引用:0點閱:164評分:下載:20書目收藏:0 大腸桿菌(Escherichiacoli,E.coli)為一種兼性厭氧性革蘭氏陰性菌，一般在溫血動物的腸道中即可見，屬於正常的腸道微生物，然而具有毒力的大腸桿菌會感染動物。

在豬，大腸桿菌可造成大腸桿菌性下痢、豬水腫病等。

藉由毒素基因(或稱毒力因子)歸類出其致病與否對於瞭解大腸桿菌的風險極為重要，然而具有毒素基因的大腸桿菌對動物的風險，需藉由生物模式來更透徹地研究。

秀麗隱桿線蟲(Caenorhabditiselegans,C.elegans)是一種生活在溫帶環境的土壤或腐敗水果中並以微生物為食的線蟲，被視為一種簡便的生物模式，常用來研究致病菌的毒力，但用於豬大腸桿菌的研究卻很少，故本研究挑選常見於豬隻大腸桿菌症的大腸桿菌毒素基因，以此檢測5株標準菌株(E.coliOP50、ATCC25922、ATCC43893、BCRC15372、ATCC35150)以及70株來自國立中興大學動物疾病診斷中心臨床微生物室自西元2016年1月1日至2017年6月30日期間所收集疑似豬大腸桿菌症感染之大腸桿菌臨床分離株。

三株標準菌株(ATCC43893、BCRC15372、ATCC35150)及15株臨床分離株(15/70,21.43%)具有上述毒素基因，其中兩株標準菌株(BCRC15372、ATCC35150)、12株具毒素基因的臨床分離株(12/15,80.00%)與5株不具毒素基因的臨床分離株(5/5,100.00%)對於秀麗隱桿線蟲具有顯著致病性。

由此可知秀麗隱桿線蟲模式具有評估ETEC(BCRC15372)、EPEC(ATCC35150)、EHEC(ATCC35150)感染的潛力，但於EIEC(ATCC43893)感染的評估效果需要進一步商榷。

此外，只具有east1基因的6株臨床分離株均對線蟲有顯著致病性，推測此結果原因為east1基因與造成宿主腸道病變有關，且秀麗隱桿線蟲的腸道構造極為發達，導致秀麗隱桿線蟲受到east1影響時容易有壽命顯著下降的結果。

兩株具有多個毒素基因的臨床分離株對秀麗隱桿線蟲不具顯著致病性，代表這些毒素基因可能具有交互作用或基因表現的程度較低而影響大腸桿菌在秀麗隱桿線蟲的致病表現。

五株不具毒素基因的臨床分離株均對秀麗隱桿線蟲有顯著致病性，代表僅以常見豬大腸桿菌症之毒素基因難以與秀麗隱桿線蟲致病性產生關連，或許需以全基因體定序找出豬臨床大腸桿菌的毒素基因，或以西方墨點法確認這些毒素基因的表現，甚至略過毒素基因部分直接以大腸桿菌症症狀明顯的豬隻進行大腸桿菌對秀麗隱桿線蟲的攻毒試驗等其他更具體的方式找出大腸桿菌在秀麗隱桿線蟲的致病性以及在豬的致病性之間的連結才能有更具體的結論。

另外，對秀麗隱桿線蟲具有顯著致病性的豬大腸桿菌在熱處理後均不會對秀麗隱桿線蟲有顯著的影響，代表這些大腸桿菌對於秀麗隱桿線蟲的有害毒素會受到熱處理的影響而失去活性，或代表這些大腸桿菌在秀麗隱桿線蟲的毒力需要大腸桿菌存活才能完整表現。

Escherichiacoli(E.coli)isafacultativeanaerobic,gram-negativebacterium.Generally,E.coliexistsnormallyintheintestineofwarm-bloodedanimals;however,thevirulentE.colicaninfectanimals.Inswine,E.colicancausecolibacillosisdiarrhea,edemadisease,etc.Thus,itisimportanttoknowthepathogenicityofE.colibydetectingitsvirulencegenes(orvirulencefactors).However,itspathogenicityinanimalsshouldbeinvestigatedthoroughlybyananimalmodel.Caenorhabditiselegans(C.elegans)isakindofnematode,livingintemperatesoilordecayingfruits.C.elegansisamajormodelorganism,commonlyusedtostudythepathogenicityofpathogenicbacteria,butthestudythatusingC.eleganstoidentifytheE.coliofswineisuntouched.Thus,5standardstrainsofE.coli(E.coliOP50,ATCC25922,ATCC43893,BCRC15372,ATCC35150)and70clinicalisolatesofE.coliinswinefromthebacteriallaborarotyintheAnimalDiseaseDiagnosticCenterofNationalChungHsingUniversityfromJanuary1st,2016toJune30th,2017werecollectedtodetectfortheE.colivirulencegeneswhicharecommonorimportantincolibacillosisofswine.Threestandardstrains(ATCC43893,BCRC15372,ATCC35150)and15clinicalisolateswithvirulencegeneswereincluded.Amongthesestrains,2standardstrains(BCRC15372,ATCC35150)and12clinicalisolates(12/15,80.00%)arestatisticallyharmfulonC.elegans.TheaboveresultsshowedthattheC.elegansassaymaybeabletoestimatethepathogenicityofETEC(BCRC15372),EHEC,EPEC(ATCC35150),buttheabilitytoevaluatethepathogenicityofEIEC(ATCC43893)needsmorestudiestobeproven.Besides,east1isthemostcommonvirulencegeneamongtheseclinicalisolates,and6clinicalisolateswithonlyeast1arestatisticallyharmfulonC.elegans,whichmaycorrelatewiththecombinationreasonsofintestinelesionsinducedbyeast1andwelldevelopedintestineinC.elegans.TwoclinicalisolateswithseveralvirulencegenesarenotstatisticallyharmfulonC.elegans,whichmeansthesegenesmayhaveinteractionwitheachotherortheexpressionofthesegenesislowthatinfluencestheresultsofC.elegans.Fiveclinicalisolates(5/5,100.00%)withoutvirulencegenesarestillstatisticallyharmfulonC.elegans,whichsuggeststhatassociationbetweenthecommonvirulencegenesinswinecolibacillosisandthepathogenicityonC.elegansislow.Inthefurture,theotherwayslikewholegenomesequencing(tofindvirulencegenes)ordirectlyusingisolatesinswinewhoseclinicalsignsofcolibacillosisareobviouscanbeusedtocorrelatewiththepathogenicityofC.elegans.Inaddition,thesedetrimentalE.colichangedtobebeneficialonC.elegansoncethroughheat-killedtreatment,whichsuggestedthatthesevirulencefactorsweresusceptibletoheat,orthevirulenceofE.colicorrelateswithitssurvivalstate. 摘要iAbstractii目次iii表目次v圖目次vi第一章文獻探討1第一節大腸桿菌之介紹1一、歷史背景1二、生長特性1第二節大腸桿菌感染症1一、致病性大腸桿菌1二、各類致病性大腸桿菌介紹2三、致病性大腸桿菌症診斷、治療、預防6四、豬隻大腸桿菌感染症8五、豬隻大腸桿菌症診斷、治療、預防10第三節大腸桿菌之毒素基因11一、腸內致病型大腸桿菌11二、腸外致病型大腸桿菌14第四節秀麗隱桿線蟲感染模式16一、歷史背景16二、生長特性及生活史16三、生物模式16第五節研究目的17第二章材料與方法18第一節細菌來源與培養18一、大腸桿菌菌株來源18二、大腸桿菌培養基及培養液18第二節大腸桿菌毒力因子之分析19一、豬大腸桿菌毒素基因選取19二、大腸桿菌毒素基因檢測19第三節秀麗隱桿線蟲之培養20一、秀麗隱桿線蟲專用培養基20二、秀麗隱桿線蟲之培養21第四節秀麗隱桿線蟲生存曲線之測定22一、大腸桿菌感染秀麗隱桿線蟲之攻毒試驗22二、熱處理大腸桿菌之秀麗隱桿線蟲攻毒試驗23第五節統計分析23第三章結果24第一節大腸桿菌毒素基因分析結果24一、大腸桿菌標準菌株毒素基因分析結果24二、臨床豬大腸桿菌毒素基因分析結果24第二節大腸桿菌之秀麗隱桿線蟲攻毒試驗24一、大腸桿菌標準菌株之秀麗隱桿線蟲攻毒試驗24二、具有毒素基因之臨床豬大腸桿菌之秀麗隱桿線蟲攻毒試驗24三、不具有毒素基因之臨床豬大腸桿菌之秀麗隱桿線蟲攻毒試驗25第三節熱處理大腸桿菌之秀麗隱桿線蟲攻毒試驗25第四節大腸桿菌在鏡檢下對秀麗隱桿線蟲外觀之影響25第四章討論27第一節大腸桿菌標準菌株及臨床豬大腸桿菌之毒素基因分析結果27第二節大腸桿菌標準菌株之秀麗隱桿線蟲攻毒試驗27第三節具有毒素基因之臨床豬大腸桿菌之秀麗隱桿線蟲攻毒試驗28第四節不具有毒素基因之臨床豬大腸桿菌之秀麗隱桿線蟲攻毒試驗29第五節大腸桿菌之秀麗隱桿線蟲攻毒試驗與熱處理組之結果比較29第五章總結31表次32圖次44參考書目54 一、中文論文1.吳迎晨(2012)。

台灣中南部屠宰場本土牛隻屠體受病原性大腸桿菌汙染之情況分析。

碩士論文，國立中興大學微生物暨公共衛生學研究所，台中，台灣。

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