(3.227.0.150) 您好!臺灣時間:2021/05/08 09:12
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:林育震
研究生(外文):Yu-Chen Lin
論文名稱:台灣北部沿岸海水中海洋弧菌之多樣性
論文名稱(外文):The Diversity of Marine Vibrios in Coastal Seawater of Northern Taiwan
指導教授:謝文陽
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:2004
畢業學年度:92
語文別:中文
論文頁數:77
中文關鍵詞:海洋弧菌
外文關鍵詞:marine vibrios
相關次數:
  • 被引用被引用:4
  • 點閱點閱:184
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
海洋弧菌是海洋生態系中數目最多、分布最廣、生理生態功能最豐富之細菌群。因此,以海洋生態系及海洋生物多樣性之研究而言,海洋弧菌多樣性之探討,實為其中不可或缺之重要環節。為探知台灣北部沿岸海水中海洋弧菌多樣性,本研究選定翡翠灣、八斗子、深澳與南雅四處沿岸樣區,自2003年11月以迄2004年2月,逐月進行海水樣本採集。利用peptone-yeast extract-glucose(PYG)平板培養基,配合嫌氣增菌培養方式,估得四處採樣地點海洋弧菌之生菌數目大致皆為102 cells/ml,其中僅有翡翠灣及南雅各有一次所測值為101 cells/ml。海水中海洋弧菌生菌數通常約占異營性細菌生菌數之十分之一。本研究四次生菌數計數之PYG平板培養系列共分離出628株細菌,其中有433株都為具發酵葡萄糖能力者。具發酵葡萄糖能力之433株分離株,根據其各種表型特性(革蘭氏反應、能否在1﹪和10﹪之NaCl濃度中生長、能否發酵inositol、D-arabinose、ducitol和xylose等醣類)檢測結果,可以分成二十三群。其中屬於第一或第二群之43株皆為革蘭氏陽性,顯然不是海洋弧菌;其餘二十一群(第三至二十三群)所含總共390株分離株,則皆具有海洋弧菌之共通表型特性。

具有海洋弧菌共通表型特性的二十一群菌株之生長都需要鈉鹽,且在3 ﹪NaCl下都能生長良好。這顯示它們都是海洋原生型細菌。這些類海洋弧菌分離株都無法在42 ℃下生長,且只有被歸類至第十五和十七群中之十四株(分別為三株和十一株)能生長於4 ℃。此意謂它們之中只有極少數耐低溫,且都為不耐熱之常溫細菌(mesophiles)。

從二十一群類海洋弧菌分離株各挑取一株進行16S rDNA定序的結果,有十二株得到近乎全部之序列,七株只定出前半段,另外兩株定序失敗。獲得完整或部分序列資料之十九株,經互相比對發現,其序列相似度介於89.1-99.2 ﹪,其中只有從第三、六、二十一和二十三群挑取之四株,互相具有97 ﹪以上之序列相似度。由此可以推斷,十九株選自不同分群之定序株,至少應可區分為十六種。

從二十一群海洋弧菌各挑取兩株(包括以上作了16S rDNA定序之二十一株),進行其16S rDNA的Dde I、Rsa I和Sca I三種內切酶之切割反應後,再就所獲酶切片段分析其限制片段長度多型性(restriction fragment length polymorphism;簡稱RFLP),結果發現三種內切酶的RFLP類型依序為一型(只有屬於不同群中兩株例外)、十七型、十七型。作了16S rDNA定序之二十一株中,只有分屬第三、六和八群之三株具有相同型之Rsa I/RFLP及Sca I/RFLP,其餘十八菌株彼此間,及它們與第三、六、八群之三株間,都在兩RFLP型中至少有一型之差異。

綜合16S rDNA之序列比對及其RFLP型分析結果,作了16S rDNA定序之二十一菌株或可進一步區分為十七種,其中挑自第三和六群之兩株可能為同種,至於挑自第八、二十一和二十三群之三株與該兩株是否同種,有待釐清。

若將選自同群內菌株兩兩比較,其Dde I/RFLP、Rsa I/RFLP及Sca I/RFLP三型完全相同者,只有第三、六和八群內之三對菌株,其餘均有一型以上差異。這顯示大部分菌群可能都不只包含一菌種。

本研究結果顯示,台灣北部沿岸海水所含海洋弧菌具有很高的種多樣性。
Marine vibrios have the most abundance of the physiological and ecological function which are most widely distributed in marine ecosystems.Thus,it''s very important to study the diversity of marine vibrios for studying the diversity of marine ecosystems.

Attempted to know the diversity of marine vibrios in coastal seawater of northern Taiwan,the seawater samples were collected every month from Fei-Zuei Bay, Badouzih, Shenao and Nan-Ya of Keelung, Taiwan during period of November 2003 to February 2004.

Plate counting values of the marine vibrios in the samples were mostly in the range of 101 to 102 cells/ml.The values usually accounted for 10﹪of the heterotrophic bacteria.628 bacterial strains were isolated from the counting plates in the study.But 433 of these bacterial strains have the ability of fermentation for glucose.They could be divided into twenty-three groups according to the results of the following tests:Gram reaction, requirement of Na+ for growth, fermentation of inositol, D-Arabinose, dulcitol and Xylose.43 strains of group I and group Ⅱ were Gram-positive.Obviously,they were not marine vibrios.390 of the twenty-one groups all have the character of marine vibrios.

From each of the twenty-one groups,we chose one strain for 16S rDNA sequence and two strains for 16S rDNA PCR-RFLP after the response by three kinds of enzyme(Dde I,Rsa I,Sca I).After analysising the results,the twenty-one groups could be further divided into seventy species.And only three pairs of group Ⅲ,group Ⅵ and group Ⅷ compelet the same after 16S rDNA PCR-RFLP.Thus,most of the groups could not contain only one specie.The study seems that it´s been exist the high diversity of species of marine vibrios in coastal seawater of northern Taiwan.
目錄

摘要…………………………………………………1

前言…………………………………………………6

材料與方法…………………………………………11

結果…………………………………………………22

討論…………………………………………………30

研究成果……………………………………………37

參考文獻……………………………………………38

圖表…………………………………………………45

附錄…………………………………………………73
林金盈 (2003). 利用菌落聚合酶連鎖反應快速檢測海水中瘡傷弧菌可行性之探討. 碩士論文. 國立台灣大學海洋研究所.

Alsina, M. & Blanch, A. R. (1993). A set of keys for biochemical identification of environmentalVibrio species. J. Appl. Bacteriol. 1993, 76, 79-85.

Baumann, P. & Baumann, L. (1984). Genus Π. Photobacterium Beijerinck 1889, 401. In Bergey''s Manual of Systematic Bacteriology. vol. 1, pp. 539-545. Edited by N. R. Krieg & J. G. Holt. Baltimore:Williams&Wilkins.

Baumann, P. & Schubert, R. H. W. (1984). Family Π. Vibrionaceae Veron 1965, 5245. In Bergey''s Manual of Systematic Bacteriology. vol. 1, pp. 516-517. Edited by N. R. Krieg&J. G. Holt. Baltimore:Williams&Wilkins.

Baumann, P., Furniss, A. L. & Lee, J. V. (1984). Genus I. Vibrio Pacini 1854, 411. In Bergey''s Manual of Systematic Bacteriology. vol. 1, pp. 518-538. Edited by N. R. Krieg&J. G. Holt. Baltimore:Williams&Wilkins.

Benediktsdottir E., Helgason S. & Sigurjonsdottir H. (1998). Vibrio spp. isolated from salmonids with shallow skin lesions and reared at low temperature. J. Fish Dis. 21, 19-28.

Blake, P. A. Weaver, R. E. & Hollis, D. G. (1980). Diseases of humans(other than cholera)caused by vibrios. Annu. Rev. Microbiol. 34, 341-367.

Brayton, P. R., P. A. West, E. Russek, & P. R. Colwell(1983). New selective plating medium for isolation of vulnificus biogroup 1. J. Clin. Microbiol. 17: 1039-1044.

Chou, C. H. & Shieh, W. Y. (2000). Vibrio vulnificus-like bacteria of cellobiose-fermentative type in seawater from the coast of north Taiwan. Acta Oceanogra. Taiwanica 38, 163-179.

Egidius, E., Wiik, R., Anderson, K., Hoff, K. A. & Hjeltnes, B. (1986). Vibrio salmonicida sp. Nov., a new fish pathogen. Int. J. Syst. Bacteriol. 36, 518-520.

Farmer, J. J., III, Hickman-Brenner, F. W., Fanning, G. R., Gordon, C. M. & Brenner, D. J. (1988). Characterization of Vibrio metschnikovii and Vibrio gazogenes by DNA-DNA hybridization and phenotype. J. Clin. Microbiol. 26, 1993-2000.

Goudie E. D. & Gow J. (1995). The taxonomic significance of the growth response to Na+ by strains of Vibrio. Can. J. Microbiol., 41: 930-935.

Hada, H. S., West, P. A., Lee, J. V., Stemmler, J. & Collwell, P. R. (1984).Vibrio tubiashii sp. Nov., a pathogen of bivalve mollusks. Int. J. Syst. Bacteriol. 34, 1-4.
Haldy, W. G. & Klontz, K. C. (1996). The epidemiology of Vibrio infections in Florida. 1981-1993.J. Infect. Dis. 173, 1176-1183.

Huq, A. & Colwell, R. R. (1995). Vibrios in the marine and estuarine environments. J. Mar. Biotechnol. 3, 60-63.

Lee, K. K., Liu, P. C. & Chuang, W. H. (2002). Pathogenesis of gastroenteritis caused by Vibrio carchariae in cultured marine fish. Mar. Biol. 4, 267-277.

MacDonell, M. T. & Colwell, R. R. (1985). The phylogeny of the Vibrionaceae, and recommendation for two new genera, Listonella and Shewanella. Syst. Appl. Microbiol. 6, 171-182.

Maeda T., Matsuo Y., Furushita M. & Shiba T. (2003). Seasonal dynamics in a coastal Vibrio community examined by a rapid clustering method based on 16S rDNA. Fisheries Sci. 2003; 69: 385-394.

M. K. Nishiguchi (2000). Temperature Affects Species Distribution in Symbiotic Populations of Vibrio spp. Appl. Environ. Microbiol. Aug. 2000, p. 3550-3555.

Ortigosa M., Garay E. & Pujalte M. J. (1993). Numerical Taxonomy of Vibrionaceae Isolated from Oysters and Seawater Along an Annual Cycle. System. Appl. Microbiol. 17, 216-225.

Sabry S. A., Ghozlan H. A. & Abou-Zeid D. M. (1996). Metal tolerance and antibiotic resistance patterns of a bacterial population isolated from sea water. J. Appl. Microbiol. 1997, 82, 245-252.

Shieh W. Y., Simidu U. & Maruyama Y. (1989). Enumeration and Characterization of Nitrogen-Fixing Bacteria in an Eelgrass(Zostera marina)Bed. Microbiol. Ecol. 18: 249-259.

Shieh, W. Y., Chen, Y. W. & Jean, W. D. (2002). Density and Phylogeny of Facultatively Anaerobic, Halophilic Bacteria that Ferment Carbohydrates with Gas Production in Coastal Seawater of Northern Taiwan. Acta Oceanogra. Taiwanica 40, 179-198.

Shieh, W. Y., Chen, Y. W., Chaw, S. M. & Chiu, H. H. (2003). Vibrio ruber sp. Nov., a red, facultatively anaerobic, marine bacterium isolated from sea water. Int. J. Syst. Evol. Microbiol., 53, 479-484.

Simidu, U., Ashino, K. & Kaneko, E. (1971). Bacterial flora of phyto and zoo-plankton in the inshore water of Japan. Can. J. Microbiol. 17, 1157-1160.

Skjermo J. & Vadstein O. (1998). Techniques for microbial control in the intensive rearing of marine larvae. Aqua. 177, 333-343.

Stackebrandt, E. & Goebel, B. M. (1994). Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst. Bacteriol 44, 846-849.

Sugita H., Matsuo N., Hirose Y., Iwato M. & Deguchi Y. (1997). Vibrio sp. Strain NM 10, Isolated from the Intestine of a Japanese Coastal Fish, Has an Inhibitory Effect against Pasteurella piscicida. Appl. Environ. Microbiol., Dec., p. 4986-4989.

Thompson F. L., Hoste B., Vandemeulebroecke K. & Swings J. (2003). Reclassification of Vibrio hollisae as Grimontia hollisae gen. Nov., comb. Nov. Int. J. Syst. Evol. Microbiol., 53, 1615-1617.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
系統版面圖檔 系統版面圖檔