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研究生:陳明裕
研究生(外文):Ming-Yu Chen
論文名稱:臺灣中部地區家禽大腸桿菌分離株抗菌劑抗藥性及分子流行病學研究
論文名稱(外文):Antimicrobial Resistance and Molecular Characteristic of Avian Escherichia coli Isolates in Middle Taiwan.
指導教授:沈瑞鴻
指導教授(外文):Jui-Hung Shien
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫學系
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:2005
畢業學年度:93
語文別:中文
論文頁數:56
中文關鍵詞:家禽大腸桿菌抗藥性
外文關鍵詞:Avian Escherichia coliResistance
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本研究於2004年5月至2005年2月間收集自中興大學疾病診斷中心罹病雞隻所分離之93株大腸桿菌,依據National Committee for Clinical Laboratory Standards ( NCCLS )之Bauer-Kirby之單一劑量瓊脂擴散試驗檢測對14至20抗菌劑的敏感性及以肉湯微量稀釋法( Broth Microdilution method )檢測3種抗菌劑的最小抑制濃度(Minimum Inhibitory Concentration, MIC)。結果中部地區大腸桿菌分離株均具多重抗藥性,而且抗藥性盛行率大多數已超過70%以上,敏感性高於50%以上之抗菌劑只賸下Colistin (93.66%)、Ceftiofur (78.43%)、Florfenicol (69.06%)、Norfloxacin (62.5%) 和Ofloxacin (50%)。本研究另收集來自相同種雞場供應小雞的二個不同肉雞場的墊料、飲水及雞隻共泄腔的53株大腸桿菌分離株,連同先前收集到之大腸桿菌分離株,以其質體相(plasmid profile)及隨機增幅多型性DNA (Random Amplified Polymorphic DNA, RAPD) 的圖型來分析大腸桿菌的分佈及流行病學。依本研究結果認為禽種間飼養環境、設備條件的優劣與大腸桿菌症的發生有關;抗藥性盛行率居高不下足見抗菌劑的使用不夠謹慎;抗菌劑使用的差異會造成禽種、地區甚至每個養禽場之間抗藥性盛行率及抗藥性模式的不同;同一病例分離株有著比較相似的抗藥性模式及質體相。除了控制禽舍環境的衛生條件可預防感染外,建議平時可以依個別禽場為單位,先行監測雞群共泄腔大腸桿菌的抗藥性模式,做為必要時防治大腸桿菌症的參考,以減緩大腸桿菌抗藥性的產生和傳播。
In this study, we collected 93 strains of Escherichia coli (E. coli) from May 2004 to February 2005 in middle Taiwan. These strains were tested the antimicrobial susceptibility with 14~20 antimicrobials by the Bauer Kirby Disk Diffusion Method and the Minimum Inhibitory Concentration (MIC) of 3 antimicrobials by broth dilution method. Consequently, all of the strains we isolated in middle Taiwan resisted many kinds of antibiotics. Furthermore, there were more than 70% strains which can resist antibiotics but only 5 kinds of antibiotics reached sensitivity over 50%, they were Colistin (93.66%), Ceftiofur (78.43%), Florfenicol (69.06%), Norfloxacin (62.5%), and Ofloxacin (50%). In addition, we chose two broiler farms that their chicks came from single breeder farm. We collected litters, waters, and cloacae swabs from these farms which were isolated 53 E. coli strains. To analyze the epidemiology of E. coli, we used Plasmid Profiling and Random Amplified Polymorphic DNA (RAPD) methods. As a result, we found that the conditions of environment and equipment closely related to the distribution of avian colibacillosis. Because the antiseptics did not use seriously, the prevalence of antibiotics-resistance strains was found at high level. The difference between antibiotics、avian species、farms and areas could induce different prevalence of antibiotics-resistance strains and infected models. We found that different strains isolated from a case had similar antibiotics resistance pattern and plasmid profiles. To sum up, by monitoring the sensitivity of antibiotics and finding out the antibiotics-resistance models, we could dwindle the transmission of E. coli.
頁數
中文摘要…………………………………………………………………………… i
英文摘要…………………………………………………………………………… ii
目次………………………………………………………………………………… iii
表次………………………………………………………………………………… v
圖次………………………………………………………………………………… vi
第一章 緒言……………………………………………………………………… 1
第二章 文獻探討………………………………………………………………… 2
第一節 家禽大腸桿菌的生化特性……………………………………… 2
第二節 家禽大腸桿菌的分型…………………………………………… 3
第三節 家禽大腸桿菌的流行病學……………………………………… 5
第四節 家禽病原性大腸桿菌的致病機制……………………………… 6
第五節 家禽大腸桿菌症………………………………………………… 8
第六節 家禽大腸桿菌的防治…………………………………………… 8
第七節 家禽大腸桿菌產生抗藥性的機制……………………………… 12
第八節 家禽大腸桿菌抗藥性的監測…………………………………… 13
第九節 隨機增幅多型性DNA(RAPD)及應用………………………… 14
第三章 材料與方法……………………………………………………………… 16
第一節 家禽大腸桿菌的來源…………………………………………… 16
第二節 家禽大腸桿菌分離及鑑定……………………………………… 16
第三節 受測大腸桿菌接種菌液的準備………………………………… 18
第四節 抗菌劑紙錠瓊脂擴散法(disk agar diffusion method)…………… 18
第五節 最小抑制濃度(minimum inhibitory concentration;MIC)檢測…… 20
第六節 質體相分析(Plasmid profiling) ………………………………… 21
第七節 隨機增幅多型性DNA指紋圖譜分析
(Random Amplified Polymorphic DNA; RAPD)………………… 21
第八節 Simpson's區別指數及相似度的應用…………………………… 23
第四章 結果……………………………………………………………………… 24
第一節 家禽大腸桿菌分離的收集……………………………………… 26
第二節 抗菌劑感受性試驗……………………………………………… 22
第三節 最小抑制濃度…………………………………………………… 33
第四節 家禽大腸桿菌質體相分析……………………………………… 34
第五節 家禽大腸桿菌隨機增幅多型性DNA指紋圖譜分析 ………… 35
第五章 討論……………………………………………………………………… 45
第六章 參考文獻………………………………………………………………… 48
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