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研究生:周青慧
研究生(外文):ChingHuei Chou
論文名稱:臺灣北部沿岸海水中類創傷弧菌之研究
論文名稱(外文):Vibrio vulnificus-like bacteria in seawater from the coast of northern Taiwan
指導教授:謝文陽
指導教授(外文):Wung Yang Shieh
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:海洋研究所
學門:自然科學學門
學類:海洋科學學類
論文種類:學術論文
論文出版年:2000
畢業學年度:88
語文別:中文
論文頁數:66
中文關鍵詞:創傷弧菌B型多黏桿菌素黏桿菌素海水浴場
外文關鍵詞:Vibrio vulnificuspolymyxin Bcolistinseawater
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創傷弧菌 ( Vibrio vulnificus ) 是廣泛分布於沿岸和河口生態系的海洋原生細菌,可經由海產食物和外傷感染人體,導致腸胃炎、蜂窩組織炎等疾病,嚴重者甚至引發致死性敗血症。以往已有不少關於海產介殼類動物體內創傷弧菌的報告,但是有關此病原菌在天然海水中分佈之調查研究仍相當有限。創傷弧菌、霍亂弧菌(Vibrio cholerae)和腸炎弧菌(Vibrio parahaemolyticus )是現知弧菌類中僅有對B型多黏桿菌素(polymyxin B)和黏桿菌素(colistin)兼具抗藥性者;該三弧菌中只有創傷弧菌具有發酵纖維二醣(cellobiose)和柳糖(salicin)之能力。本研究綜合創傷弧菌的這些特性,改良了一前人使用過之纖維二醣-B型多黏桿菌素-黏桿菌素培養基(cellobiose-polymyxin B-colistin medium;簡稱CPC培養基),並設計出一柳糖-B型多黏桿菌素-黏桿菌素培養基(salicin-polymyxin B-colistin medium;簡稱SPC培養基),以進行台灣北部沿岸海水中對象菌的培養、計數、分離和鑑定之探討,並將能在此二培養基上生長者,一概稱之為類創傷弧菌(Vibrio vulnificus-like bacteria)。調查結果顯示,沙崙、白沙灣、金山和翡翠灣等海水浴場海水以及關渡和竹圍紅樹林區之河口水中,類創傷弧菌的生菌數多不出100~103 cells / ml。利用CPC與SPC培養基進行最可能數計數(most-probable-number counts;簡稱MPN counts)所估算得到的菌數無明顯差異。本研究一共分離出66株類創傷弧菌,根據生理和生化等表型特徵之檢測結果,可將這些分離株區分為六群,包括四群具需鹽性和葡萄糖發酵性的弧菌科海洋細菌,及兩群不具需鹽性的葡萄糖發酵性陸源細菌。隸屬弧菌科之四菌群中,只有含21株之一群(佔全部分離株之32%)的代表株與創傷弧菌,具有97%以上之16S rDNA序列相似性。利用CPC和SPC兩培養基進行沿岸及河口水域中創傷弧菌之篩選實驗,結果顯示,該菌在二培養基上之出現率皆以採自沿岸之水樣為高,顯示該二培養基對沿岸地區海水中創傷弧菌的篩選力較佳。本研究結果亦顯示,適度改變CPC與SPC二培養基中B型多黏桿菌素和黏桿菌素的濃度,應能提昇其對創傷弧菌篩選之有效性。
Vibrio vulnificus is an autochthonous marine bacterium which is widely distributed in estuarine and coastal ecosystems; seafood and water can be vectors for transmission of this pathogenic species to humans. Its infection might result in a variety of clinical syndromes, including gastroenteritis, cellulitis, and septicemia. There are many reports for abundance of V. vulnificus in shellfish, but rare studies for its distribution in seawater. V. vulnificus, Vibrio cholerae and Vibrio parahaemolyticus are the only species in the genus Vibrio, which are known to be resistant to both polymyxin B and colistin. Among the three species only V. vulnificus can ferment cellobiose and salicin. According to the characteristics found in V. vulnificus, we used a modified cellobiose-polymyxin B-colistin medium ( CPC medium ) and developed a salicin-polymyxin B-colistin medium ( SPC medium ) for cultivation, enumeration, isolation and identification of V. vulnificus-like bacteria in seawater samples from the coast of northern Taiwan. Most MPN counting values of these bacteria were in the range of 100 to 103 cells / ml for coastal water samples from Shalun, Paishawan, Chinshan and Fetsueiwan, and for estuarine water samples from Kuantu and Chuwei. The counting values on CPC and SPC media were not significantly different. Sixty-six strains of V. vulnificus-like bacteria isolated in this study were divided into six groups according to their phenotypic characteristics. All the isolated bacteria fermented glucose while only four groups included halophilic isolates. Only the representative strains included in the greatest group ( 21 isolates ) showed levels of 16S rDNA sequence similarity more than 97% with V. vulnificus. Higher incidence of V. vulnificus were observed on coastal water than on estuarine water samples ( 60-100% : 0-40% ) by using the present cultivation methods. Appropriate modifications of the concentration of polymyxin B and colistin in CPC and SPC media would enhance the selectivity of these media for V. vulnificus.
目 錄
摘 要. . . . . . . . . . . . . . . . . . . . . . . . . . 1
英文摘要. . . . . . . . . . . . . . . . . . . . . . . . . 3
前 言. . . . . . . . . . . . . . . . . . . . . . . . . . 5
材料與方法. . . . . . . . . . . . . . . . . . . . . . . .10
結 果. . . . . . . . . . . . . . . . . . . . . . . . . .19
討 論. . . . . . . . . . . . . . . . . . . . . . . . . .25
參考文獻. . . . . . . . . . . . . . . . . . . . . . . . .28
附 表. . . . . . . . . . . . . . . . . . . . . . . . . .34
附 圖. . . . . . . . . . . . . . . . . . . . . . . . . .44
附 錄. . . . . . . . . . . . . . . . . . . . . . . . . .62
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