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研究生:陳秀芬
研究生(外文):Hsiu-Fen Chine
論文名稱:以脈衝式電泳分析犬沙門氏桿菌之流行病學研究
論文名稱(外文):Epidemiologic Study of Canine Salmonellosis by
指導教授:廖明輝廖明輝引用關係
指導教授(外文):Ming-Huei Liao
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:獸醫學系所
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:2008
畢業學年度:96
語文別:中文
論文頁數:69
中文關鍵詞:沙門氏桿菌脈衝式電泳藥物敏感性試驗
外文關鍵詞:Salmonellapulsed-field gel electrophoresisantibiotic susceptibility test
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本研究於2006年針對高雄地區選定12家動物醫院、8家寵物店及高雄市動物收容處所隨機採檢糞材,並參照世界動物衛生組織(International Office of Epizootics;OIE)分離流程,進行沙門氏桿菌的分離鑑定。共採集1,828件糞材樣本進行鑑定,合計分離出256件沙門氏桿菌,其中自動物醫院分離出的陽性株58件(22.7%;58/256)、寵物店18件(7.0%;18/256),動物收容中心180(70.3%;180/256)件;分離率以動物收容中心陽性樣本數為最高。
沙門氏桿菌屬於革蘭氏陰性菌,具人畜共通傳染性,可造成人類嚴重的腸胃炎、菌血症及傷寒等疾病與經濟損失,迄今仍為世界各國重要的公共衛生問題。沙門氏桿菌屬於典型的腸道病原菌,經口為最普遍的感染途徑,主要是吃到污染病原的食物,感染後病原可從糞便排出;某些無臨床症狀的感染動物成為潛在性的帶菌者,由糞便散播沙門氏桿菌污染環境造成其他動物及人類的感染發病。
隨著生活型態的改變,圈養寵物的人口逐年攀升,由家庭寵物來源感染的沙門氏桿菌症成為新的關注焦點;根據研究結果顯示,自寵物來源的隨機採樣,犬隻感染沙門氏桿菌的盛行率為14%;進一步以脈衝式電泳(pulsed-field gel electrophoresis, PFGE)基因圖譜與傳統血清型試驗鑑定分析沙門氏桿菌分離株,共鑑定出12種血清型,其中以S. Albany(76.2%)及S. Newport(12.2%)居多;藥物敏感性試驗中顯示,藥物抗藥性比例以tetracycline(85.7%)、ampicillin(78.2%)及sulfamethoxazole- trimethoprim(78.2%)三種藥物最高;依據以上結果,顯示與人們一起生活的寵物,有可能成為人類沙門氏桿菌症的潛在感染來源,未來藉由以上資料,不但可提供台灣民眾於飼養寵物的衛教資訊及臨床獸醫師於犬隻施用抗生素藥物時之參考,並可提供未來進行沙門氏桿菌感染之風險評估資料之依據。
The experiment was conducted in the four seasons in 2006. The 1,828 fecal samples was collected from the 12 randomly selected veterinary clinics, 8 pet shops and animal shelter in Koahsiung city. Salmonella spp. isolation and identification. Were based on the standard protocol of International Office of Epizootic (OIE),. Two hundred and fifty six from all collected samples were Salmonella positive, 58 (22.7% : 58/256) from veterinary clinics, 18 (7.0% : 18/256) from pet shops and 180 (70% : 180/256) from the animal shelter.
Salmonella spp., a gram-negative organism, could cause animal and human serious gastroenteritis such as bacteremia and bacillus typhi is considered as one of the important issue of zoonosis that could cause economic catastrophe all around the world. Salmonella spp, as a classical gastrointestinal pathogen, infects hosts mainly via the face-oral route, especially from the contaminated food. Pathogen could be found in the feces of the infected hosts which didn’t show any signs clinically. The patent hosts and serves as potential carriers. Fecal transmission pathway provides the important route for Salmonella spp. infection of animals and human beings from the eco-environment.
Salmonella infection in pet animals becomes an important public issue with the life-style changing and the increasing population with pets. As the data from the survey shows that the prevalence rate of salmonellosis in pet dogs and cats and indicates that pets have become an important source worthy of more concern and further study for Salmonella. Serotpying and PFGE pattern results revealed that the isolates comprised of 12 serotypes of Salmonella. Out of all isolates, S. Albany(76.2%) had the highest percentage followed by S. Newport(12.2%). Most of the strains tested were resistant to tetracycline(85.7%), ampicillin(78.2%)and sulfamethoxazole- trimethoprim(78.2%). On the basis of these observations, it was concluded that dog as pets could be the possible source of Salmonella infection for humans, and may eventually threat human public health. The results useful for health education in people who raise or will keep dog as pets. Future risk assessment of Salmonella infection for humans is needed in Taiwan.
目 錄
中文摘要……………………………………………………I
Abstract…………………………………………………III
誌謝…………………………………………………………Ⅴ
目錄…………………………………………………………Ⅵ
圖表目錄……………………………………………………VIII
第1章 前言……………………………………………1
第2章 文獻回顧………………………………………4
2.1 沙門氏桿菌的歷史背景與其命名……………4
2.2 沙門氏桿菌的特性……………………………5
2.3 沙門氏桿菌的傳染途徑………………………6
2.4 沙門氏桿菌的疾病與致病性…………………6
2.5 治療……………………………………………8
2.6 流行性…………………………………………8
2.7 沙門氏桿菌之分離……………………………9
2.8 沙門氏桿菌血清凝集鑑定……………………10
2.9 沙門氏桿菌的分型……………………………11
2.10 脈衝式電泳之介紹……………………………12
2.10. 1脈衝式電泳判別的準則……………………14
2.11 沙門氏桿菌的抗藥性情形……………………14
第3章 材料與方法……………………………………16
3.1. 檢體來源………………………………………16
3.2 培養基與試劑…………………………………16
3.2.1培養基………………………………………………16
3.2.2試劑…………………………………………………16
3.3 藥物敏感性試驗使用藥片……………………18
3.4 實驗設備與儀器………………………………18
3.5 沙門氏桿菌採樣及分離………………………19
3.5.1採樣方法……………………………………………19
3.5.2沙門氏桿菌分離與純化……………………………19
3.5.3沙門氏桿菌生化試驗………………………………20
3.5.4沙門氏桿菌抗血清之確認…………………………21
3.5.5菌種保存……………………………………………21
3.6 脈衝式電泳……………………………………21
3.7 血清凝集試驗…………………………………23
3.8 藥物感受性試驗………………………………24
3.9 統計分析………………………………………25
3.10 問券調查………………………………………25
第4章 結果……………………………………………26
4.1 沙門氏桿菌分離結果…………………………26
4.2 脈衝式電泳分析………………………………29
4.3 血清型別鑑定結果……………………………30
4.4 藥物感受性試驗之結果………………………36
4.5 問卷調查之結果………………………………37
第5章 討論……………………………………………50
參考文獻…………………………………………………55
附錄………………………………………………………66
作者簡介…………………………………………………69
表目錄

表. 1 沙門氏桿菌陽性檢出率與犬隻感染沙門氏菌之相關危險因子比較………………………………………………28
表. 2 收容所犬隻沙門氏桿菌陽性檢出率與季節之比較………………………………………………29
表. 3 犬隻沙門氏桿菌之血清型及其來源之比較………………………………………………34
表. 4 沙門氏桿菌血清型別與犬隻來源之比較………………………………………………35
表. 5 不同來源犬隻所分離之S. Albany抗生素感受性試驗比較………………………………………………40
表. 6 不同來源犬隻所分離之S. Newport抗生素感受性試驗比較………………………………………………41
表. 7 犬隻沙門氏桿菌抗生素感受性試驗………………………………………………42
表. 8 犬隻沙門氏桿菌抗生素感受性試驗(續)……………………………………………… 43
表. 9 動物醫院沙門氏桿菌各血清型之抗藥性比例…………………44
表. 10 寵物店沙門氏桿菌各血清型之抗藥性比例……………………45
表. 11 收容所沙門氏桿菌各血清型之抗藥性比例……………………46
表. 12 犬隻沙門氏桿菌多重抗藥性之份………………………………47
表. 13 犬隻沙門氏桿菌多重抗藥性之分佈(續)……………………48
圖目錄

圖.1 PFGE patterns of XbaΙ-diggested chormosomal DNA of Salmonella isolates and 1 type strain H9812………………30
圖.2 犬沙門氏桿菌血清型別鑑定結果分佈圖…………………31
圖.3 動物醫院犬沙門氏桿菌血清型分佈圖……………………32
圖.4 寵物店犬沙門氏桿菌血清型分佈圖………………………32
圖.5 動物收容中心犬沙門氏桿菌血清型分佈圖………………33
圖.6 不同來源犬隻所分離之S. Albany抗生素感受性試驗比較分佈圖………………………………………………………39
圖.7 人與犬隻沙門氏桿菌之血清型S. Albany、S. Newport及S. Bareilly之樹狀分析圖……………………………49
附錄

表.14 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…………………………………………………………66
表.15 單價O抗原對應H抗原快速檢索表……………67
表.16 單價O抗原對應H抗原快速檢索表 (續)……68
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