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  • 學位論文

利用分子技術檢測畜禽產品中之沙氏桿菌

Detection of Salmonella in Animal products by Molecular Techniques

指導教授 : 廖明輝

摘要


沙氏桿菌( Salmonella )為重要人畜共通傳染疾病,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,由糞便散播沙氏桿菌污染環境造成其他動物及人類的感染發病,少數血清型具有專一性,對於人類會造成嚴重腸胃炎、菌血症及傷寒等疾病,受污染之家畜禽肉類及其相關製品為亦是造成人類沙氏桿菌症的主要來源,此病造成經濟損失及為害人類甚大,迄今仍為世界各國重要的公共衛生問題。 本實驗於2009-2010年針對市售畜禽產品所分離沙氏桿菌以分子檢測進行分析,根據研究顯示以基因內轉錄間隔區 ( ITS ) 作為引子之聚合酶連鎖反應( Polymerase chain reaction, PCR ) 檢測沙氏桿菌,結果敏感性100% 呈現陽性反應,另以脈衝式膠體電泳( pulsed-field gel electrophoresis, PFGE )基因圖譜分析沙氏桿菌分離株,共鑑定出16種血清型,血清型以S. Enteritidis、S. Schwarzengrund及S. Typhimurium為最多,其中豬肉分離血清型以S. Typhimurium及S. Derby為多,雞肉分離血清型以S. Enteritidis及S. Schwarzengrund居多,加工製品分離血清型以S. Typhimurium居多,針對分離血清型做抗生素敏感試驗,發現對Tetracycline、Nalidxic acid與Streptomycin具有很高的抗藥性,在豬肉中則發現此菌對Tetracycline與Streptomycin抗藥性較高,在雞肉上中發現此菌對Nalidxic acid、Tetracycline、Trimethoprim-sulfamethoxazole與Amoxicillin抗藥性較高,在加工製品中發現此菌對Tetracycline、Trimethoprim-sulfamethoxazole、Nalidxic acid與Streptomycin抗藥性較高,不同來源之多重抗藥性及抗藥性基因亦不同,且多重抗藥性基因存在再次顯示抗生素濫用之問題,針對基因圖譜與抗藥性之間相關性仍需相關更多分析方能證明其與疾病及抗藥性相關性,以上研究資訊可提供作為沙氏桿菌防治之參考,以降低沙氏桿菌之污染。

並列摘要


Salmonella infection is a very important zoonosis that causes human and animal salmonellosis. Salmonella spp. are typical gastrointestinal pathogens which infect host via the oral-fecal route by contaminated food. Some asymptomatic hosts as reservoir because of Salmonella spp. could spread and then infected humans and animals. Some serotypes have host-specifity. It caused the serious gastroenteritis, enteric fever ( typhoid fever ) and septicemia in infected host. Most cases of human salmonellosis are associated with the consumption of contaminated food products such as meat, poultry, eggs, and dairy products. It’s still a big problem in public food safety, which may lead to health and economic loss. The current study was conducted to analyze the epidemiology of Salmonella spp. in pork, chicken, processing products and duck. Samples were collected on the National Animal Industry Foundation during 2009–2010, the internal transcribed spacer(ITS) region of 16S-23S rRNA as PCR primer detection target. The PCR sensitivity results revealed 100% positive. In addition, using pulsed-field gel electrophoresis ( PFGE ) to analyze Salmonella serotypes and genetic patterns, the results indicated that there are 16, mostly S. Typhimurium, S. Enteritidis and S. Schwarzengrund. The two common serovars were S. Typhimurium and S. Derby in pork. S. Enteritidis and S. Schwarzengrund were common serovars in chicken. S. Typhimurium was the most common serovars in processing products. Antimicrobial susceptibility testing showed that most of the serotypes strains were resistant to Tetracycline, Nalidxic acid and Streptomycin. The resistant drugs in the pork are Tetracycline and Streptomycin, in the chicken are Nalidxic acid, Trimethoprim-sulfamethoxazole, Tetracycline, and Amoxicillin, in the processing products are Trimethoprim -sulfamethoxazole , Tetracycline, Nalidxic acid and Streptomycin. The current data illustrated the different multidrug resistant and resistance genes by PFGE and antimicrobial susceptibility testing, suggested that inappropriate use of antibiotics might be the main cause multidrug- resistance in Salmonella spp. In conclusion, our results could be considered as the reference in Salmonella spp. prevention from the public health aspect and be eventually to reduce outbreaks and contaminations in food products due to Salmonella.

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