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研究生:蔡宇馨
研究生(外文):Yu-Shin Tsai
論文名稱:中部地區產蛋雞與帶殼蛋之沙門氏菌監測調查
論文名稱(外文):Investigation of Salmonella in hens and shell-eggs in central Taiwan
指導教授:陳德勛
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫公共衛生學研究所
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:2007
畢業學年度:96
語文別:中文
論文頁數:96
中文關鍵詞:沙門氏菌蛋雞雞蛋抗生素抗藥性
外文關鍵詞:SalmonellaheneggAntibiotic resistant
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依據國家衛生研究院發表,台灣主要以Salmonella Typhimurium、Salmonella Enteritidis、Salmonella Stanley、Salmonella Agona、Salmonella Albany、Salmonella Schwarzengrund、Salmonella Newport、Salmonella Choleraesuis、Salmonella Derby及Salmonella Weltevreden等為人類常見感染的血清型。沙門氏菌症 (Salmonellosis) 爲重要的人畜共通傳染病,美國每年約有140萬人以上感染非傷寒性沙門氏菌症,造成800 – 4000 件死亡案例,以公共衛生的觀點,家禽感染沙門氏菌最易引起人們食物中毒的可能。本研究藉由調查中部地區蛋雞的沙門氏菌之陽性率,同時也對生鮮的雞蛋進行檢測,所得到的沙門氏菌菌株再進行抗生素的抗藥性試驗、血清分型及相關的危險因子分析,藉此提供未來擬訂沙門氏菌控制計畫的參考。採樣自蛋雞卵巢與十二指腸部份,總計採取990隻蛋雞,雞隻的陽性率為25.5 % (252/990);沙門氏菌的樣本總陽性檢出率為18.2 % (359/1970),卵巢的沙門氏菌陽性檢出率為蛋雞十二指腸沙門氏菌陽性檢出率的1.4倍 (95 % C.I 1.09 ~ 1.73)。夏、秋兩季溫度較高進而陽性率也跟著提高,秋季的沙門氏陽性檢出風險率是春季的2.8倍(95 % C.I 1.9 ~ 4.0)。雞蛋樣本部份並未檢測到任何沙門氏菌陽性。血清分型結果以Salmonella Enteritidis血清型為最多,佔總數的91.6 %(329/359),所有菌株的35.5 %(128/359)不具抗藥性,但全都是Salmonella Enteritidis血清型,其餘菌株主要是對Colistin及Tetracycline具有較高的抗藥性。在多變相邏輯式回歸分析中,在本次實驗得到在牧場中飼養的規模、消毒的次數、環境條件、周圍是否存在其他牧場、牧場的環境、飼主平均每日在牧場內工作的時數都會影響蛋雞牧場中沙門氏菌的陽性率。以上的研究資訊將有助於發展沙門氏菌的防治策略,以減低人類感染疾病的風險及提高農產品出口競爭力。
It is reported according to NHRI ( National Health Research Institutes) that the majority of human Salmonella cases in Taiwan are caused by serotypes of Salmonella Typhimurium、S. Enteritidis、S. Stanley、S. Agona、S. Albany、S. Schwarzengrund、S. Newport、S. Choleraesuis、S. Derby and S. Weltevreden. Salmonellosis is an important zoonosis worldwide. It was estimated that 1.4 million human cases with non-typhoidal Salmonella infections, and probably 800-4000 related deaths occur in the United States annually. In terms of public health, avian salmonellosis is a hazard to cause food poisoning cases in humans. The objectives of this study were 1) to investigate the positive rate of salmonellosis of hens and fresh eggs in central Taiwan; 2) to idethntify the antimicrobial resistance, serotyping, and associated risk factors of the Salmonella isolates; 3) to provide a protocol for control of Salmonellosis. A total of 990 samples were collected from chickens’ ovaries and duodenums. The Salmonella - positive rate of these chickens was 25.5%(252/990) and of all specimens was18.2% (359/1970). The percentage of Salmonella-positive specimens of ovaries was 1.4 times more than duodenums (95 % C.I 1.09 ~ 1.73). A significantly higher percentage of Salmonella-positive specimens was identified in summer and fall accompanying higher temperature. The Salmonella-positive specimens in fall is 2.8 times more than summer (95 % C.I 1.9 ~ 4.0). None samples of the eggs are salmonella-positive. In the event of serotyping, Salmonella Enteritidis was the most common serotype accounting for 91.6 %(329/359)of Salmonella isolates. And 35.5 %(128/359)of the isolates which were all Salmonella Enteritidis were not antimicrobial resistant . The other isolates are more resistant to Colistin and Tetracycline. Multivariate Statistical Analysis indicated that magnitude of rearing in farms, the frequencies of disinfection, the neighborhood, and the working hours of the farmer were all risk factors for Salmonella preavlance in hens farms. The above information will be helpful for the development of effective strategies for prevention and reducing the risk of human Salmonella infection to strength the competence for exporting agricultural products.
中文摘要……………………………………………………………………………i
英文摘要……………………………………………………………………………ii
目次…………………………………………………………………………………iii
表目次………………………………………………………………………………vi
圖目次………………………………………………………………………………viii
第一章 前言………………………………………………………………………1
第二章 文獻探討…………………………………………………………………3
第一節 沙門氏菌……………………………………………………………3
一、 沙門氏菌簡介………………………………………………………………3
二、 沙門氏菌的歷史……………………………………………………………3
三、 沙門氏菌的生化特性………………………………………………………4
四、 人之沙門氏菌感染…………………………………………………………5
五、 家禽之沙門氏菌感染………………………………………………………7
六、 其他動物之沙門氏菌感染…………………………………………………8
七、 沙門氏菌之感染途徑………………………………………………………9
八、 沙門氏菌與公共衛生………………………………………………………10
九、 沙門氏菌之抗藥性…………………………………………………………11
第二節 沙門氏菌的命名與分類…………………………………………………13
一、 沙門氏菌的命名……………………………………………………………13
二、 沙門氏菌的抗原構造………………………………………………………14
三、 沙門氏菌的血清型分類……………………………………………………15
第三節 沙門氏菌之分離…………………………………………………………16
第四節 沙門氏菌之鑑定法………………………………………………………21
一、 血清學試驗…………………………………………………………………21
二、 生化型試驗…………………………………………………………………21
三、 細菌快速鑑定系統…………………………………………………………22
第五節 快速偵測沙門氏菌………………………………………………………23
一、 傳統方式的改良……………………………………………………………23
二、 非分離性質的快速偵測技術………………………………………………25
第六節 危害分析與重要管制點…………………………………………………25
一、危害分析與重要管制點(HACCP)的起源…………………………………26
二、危害分析與重要管制點(HACCP)的目的…………………………………27
三、 HACCP系統控制沙門氏菌污染之應用…………………………………28
第三章 材料與方法………………………………………………………………30
第一節 實驗的背景與目的………………………………………………………30
第二節 實驗材料………………………………………………………………31
一、 蛋雞採樣……………………………………………………………………31
二、 雞蛋採樣……………………………………………………………………31
第三節 樣本實驗室處理…………………………………………………………32
一、 蛋雞臟器部份…………………………………………………………32
二、 雞蛋內容物部份………………………………………………………33
三、 雞蛋表面部份………………………………………………………33
第四節 沙門氏桿菌之培養與分離………………………………………………34
第五節 沙門氏菌的血型鑑定………………………………………………………35
一、 O抗原玻片凝集試驗…………………………………………………35
二、 H抗原試管凝集試驗…………………………………………………36
三、 H抗原濾紙片相誘導…………………………………………………37
四、 血清型對照確認……………………………………………………38
第三節 抗生素耐受性試驗………………………………………………………38
第四節 統計與分析………………………………………………………………39
第四章 結果………………………………………………………………………41
第一節 樣本來源及數量………………………………………………………41
第二節 蛋雞來源樣本之沙門氏菌分離結果……………………………………42
第三節 雞蛋來源樣本之沙門氏菌分離結果……………………………………43
第四節 樣本檢出之沙門氏菌血清群及血清型的分佈…………………………43
第五節 蛋雞樣本之沙門氏菌株藥物耐受性試驗結果…………………………44
第六節 蛋雞沙門氏菌陽性之相關危險因子分析………………………………45
第五章 討論………………………………………………………………………50
第一節 蛋雞來源樣本的沙門氏菌陽性檢出情形………………………………50
第二節 雞蛋來源樣本的沙門氏菌陽性檢出情形………………………………51
第三節 蛋雞樣本分離到的沙門氏菌株以及藥物受性試驗結果………………53
第四節 蛋雞沙門氏菌陽性之相關危險因子分析………………………………55
第六章 結論………………………………………………………………………57
參考文獻……………………………………………………………………………90
附錄…………………………………………………………………………………97
表一 沙門氏桿菌生化反應特性……………………………………………………58
表二 利用生化特性區分沙門氏桿菌種別與亞種…………………………………59
表三 於1996至2000英國威爾斯郡病患分離的副傷寒型沙門氏桿菌
多重抗藥性的發生率……………………………………………………60
表四 美國CDC採用的沙門氏桿菌命名法,2000…………………………………61
表五 Salmonella biochemical and serotype test……………………………………62
表六 沙門氏菌菌體抗原測試及鞭毛抗血清檢測對照表…………………………63
表六 沙門氏菌菌體抗原測試及鞭毛抗血清檢測對照表(續)…………………64
表七 依抗生素感受性試驗抑制圈大小結果判定菌株抗藥性之標準………65
表八 蛋雞牧場每月沙門氏菌陽性檢出結果表…………………………………66
表九 二十個牧場蛋雞沙門氏菌陽性樣本檢出率及其依照不同臟器來源與季節之
比較結果……………………………………………………………………67
表十 二十個牧場蛋雞沙門氏菌陽性樣本檢出率依照不同牧場來源比較結
果……………………………………………………………………………68
表十一 蛋雞來源樣本來源之沙門氏菌血清群分佈結果…………………………69
表十二 蛋雞來源樣本之沙門氏菌菌株血清分型結果……………………………70
表十三 蛋雞樣本之沙門氏菌株藥物耐受性比較…………………………………71
表十四 蛋雞來源樣本之沙門氏菌多重抗藥性之分佈……………………………72
表十五 二十個牧場蛋雞沙門氏菌陽性樣本檢出率與使用不同品牌的飼料分類結
果表…………………………………………………………………………73
表十六 二十個牧場蛋雞沙門氏菌陽性樣本檢出率依照不同飼養規模比較結
果表………………………………………………………………………74
表十七 二十個牧場蛋雞沙門氏菌陽性樣本檢出率依照每月不同的消毒次數比較
    結果表………………………………………………………………………75
表十八 二十個牧場蛋雞沙門氏菌陽性樣本檢出率與不同環境條件分類結果
   表…………………………………………………………………………76
表十九 二十個牧場蛋雞沙門氏菌陽性樣本檢出率與飼主於牧場不同工作時數分
類結果表……………………………………………………………………77
表二十 二十個牧場蛋雞沙門氏菌陽性樣本檢出率依照牧場是否接鄰大馬路的比
較結果表………………………………………………………………78
表二十一 二十個牧場蛋雞沙門氏菌陽性樣本檢出率依照牧場周圍五十公尺是否
有其他牧場的比較結果表………………………………………………79
表二十二 S. Enteritidis 感染之危險因子多變項模式分析結果…………………80
表二十三 S. Enteritidis 感染之危險因子多變項模式分析結果…………………81
圖目次
頁次
圖一 雞蛋放置於蛋盤上,使每一個雞蛋比白可單獨站立…………………………82
圖二 沙門氏菌培養分離鑑定流程圖………………………………………………83
圖三 MSRV檢測培養沙門氏菌之情形………………………………………………84
圖四 沙門氏菌培養於XLD agar上之生長情形……………………………………85
圖五 沙門氏菌之o抗原玻片凝集試驗………………………………………………86
圖六 沙門氏菌h抗原試管凝集試驗,菌液加入抗血清……………………………87
圖七 沙門氏菌h抗原試管凝集試驗,水浴中測試凝集反應………………………88
圖八 於TSA培養基斜線部份刮除1cm寬…………………………………………89
圖九 進行沙門氏菌誘導時,於TSA培養基上使用濾紙搭成渡橋…………………90
圖十 沙門氏菌h抗原濾紙片相誘導培養法………………………………………91
圖十一 紙錠擴散法抗生素敏感性試驗藥片抑制圈情形…………………………92
圖十二 蛋雞牧場每月沙門氏菌陽性檢出結果曲線圖……………………………93
圖十三 不同季節與沙門氏菌陽性率曲線圖………………………………94
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