臺灣博碩士論文加值系統

摘 要
腸炎弧菌（Vibrio parahaemolyticus）是許多亞洲國家主要的食物中毒致病菌，但在1996前並無特定血清型的感染情形。自1996年印度出現特殊血清型O3:K6的食物中毒案件，之後新型菌株在亞洲許多城市蔓延，台灣也在其中。為了瞭解新O3:K6型大流行的特性，本研究從乳鼠致死實驗、Lipopolysaccharides（LPS）、蛋白質圖譜以及遺傳特性三方面著手，比較新O3:K6、舊O3:K6、環境菌株以及類新O3:K6四類菌株的差異。其中類新O3:K6是與新O3:K6具有相似遺傳特性但卻不同血清型的菌株，如O4:K68或O1:KUT等。
在乳鼠致死實驗中，除了一株新O3:K6（N5）的致死時間較長外，其他新O3:K6、舊O3:K6、環境菌株以及類新O3:K6並無顯著差異。腸毒性的實驗與致死實驗所得的結果一致，各類菌株並無差異。結果顯示所有新O3:K6和其他腸炎弧菌菌株都具有rough型的LPS，而所有菌株LPS圖譜分析結果皆相同。總體蛋白質圖譜分析亦同，不論是Coomassie blue stain 或 Sliver stain皆顯示新O3:K6與其他腸炎弧菌的圖譜一致。
無論是乳鼠致死實驗或蛋白質圖譜分析皆無法看出新O3:K6有別於舊O3:K6的特殊性，而其他文獻僅在遺傳特性分析上能找到新O3:K6與舊O3:K6的顯著差異。於是本研究放大各類菌株部分之toxR 、 laf 及進行兩序列分析。在toxR分析，新O3:K6與舊O3:K6有兩個鹼基的差異。特別的是新O3:K6與類新O3:K6的100%相似。而在laf分析，發現新O3:K6與舊O3:K6有一個鹼基的差異。這些鹼基位置的發現，提供了一個研究新O3:K6致病特性的新方向，或者可作為發展鑑定新O3:K6的有效方法。

ABSTRACT
Vibrio parahaemolyticus is an important foodborne pathogen in many Asian countries, but no dominant serovars were involved in food poisonings until the appearance of the new O3:K6 strains in India in 1996. The pandemic spread of these new O3:K6 strains has since occurred, involving other Asian countries including Taiwan. In order to identify special factors associated with these pandemic O3:K6 strains, the infant mouse lethality, lipopolysaccharide, protein profiles, and genetic assay of new O3:K6, old O3:K6, new O3:K6-like and environmental strains of V. parahaemolyticus were examined in this study. New O3:K6-like strains were genetically similar to the new O3:K6 strains, but with different serovars, such as the O4:K68 and O1:KUT.
Most of the selected new O3:K6 strains showed similar mouse lethality like old O3:K6, new O3:K6-like and environmental strains, but strain N5 was the least virulent one. The enterotoxigenicity of these strains were also similar. The lipopolysaccharide analysis identified rough type in the selected strains belonging to the new O3:K6 and other categories, and the LPS patterns of these strains were similar. Moreover, total protein profile of new O3:K6, old O3:K6 and new O3:K6-like strains were also similar as determined by both of Coomassie blue and Sliver stainings。
Partial seguences of toxR and laf of these strains were amplified and sequenced in this study. Comparing with the toxR sequences, there were two base positions changes in the new and old O3:K6 strains. Especially, toxR sequences of new O3:K6 and new O3:K6-like were 100% identical. Comparing with the laf sequences, the eight sequences were highly diverse. Difference in one base position was found in the new and old O3:K6 strains. These results may provide new approach to study the factors involved in the spreading of these pandemic strains, or from which new molecular methods can be developed for the specific detection of these new O3:K6 strains.

目 錄
誌謝……………………………………………………………………Ⅰ
目錄……………………………………………………………………Ⅱ
圖表目錄………………………………………………………………Ⅴ
中文摘要………………………………………………………………Ⅵ
英文摘要………………………………………………………………Ⅷ
第一章、緒 論 1
第一節、腸炎弧菌的特性 1
1.1. 一般特性及分類特性 1
1.2. 抗原性 2
1.3. 生長條件 2
1.4. 分佈狀況 4
第二節 腸炎弧菌與食物中毒 4
2.1 致病情形 5
2.2. 致病因子 5
2.2.1. 熱穩定溶血素（Thermostable direct hemolysin, TDH） 5
2.2.2. 類熱穩定溶血素（TDH-related hemolysin, TRH） 6
2.2.3. TDH/I 7
2.2.4. 致死毒素（Lethal toxin） 7
2.2.5. 吸附因子 7
2.2.6. 熱穩定細胞毒素 8
2.2.7. 血球凝集素 8
2.2.8. 內毒素 9
2.3. 致病菌株之偵測 10
2.4. 血清型與致病之關係 12
第三節 新O3：K6型腸炎弧菌的研究 13
3.1. 遺傳變異 13
3.2. 毒力因子 14
3.3. 對抗生素敏感特性 15
3.4. 對環境壓力的耐受性 15
3.5. 絲狀噬菌體 16
3.6. 其他血清型流行菌株 17
第二章、實驗目的與架構 18
第一節、實驗目的 18
第二節、實驗架構 18
2.1. 新O3:K6型腸炎弧菌的毒性 18
2.2. 蛋白質圖譜分析 19
2.3. 遺傳特性 19
第三章、材 料 與 方 法 21
第一節 菌株的選擇 21
第二節 藥品與器材 21
第三節 培養基成分 23
第四節 緩衝溶液配方 23
第五節 實驗條件與步驟 36
3.5.1. 菌株的保存與培養 36
3.5.2. 毒性分析 36
3.5.2.1. 動物致死毒性以及腸毒性 36
3.5.2.2. LPS差異 37
3.5.3. 總體蛋白質圖譜分析 40
3.5.3.1. 總體蛋白質萃取 40
3.5.3.2 蛋白質定量 41
3.5.3.3. .SDS-PAGE電泳 41
3.5.3.4. .Coomassie blue & silver staining 42
3.5.4. 遺傳差異 42
3.5.4.1. 染色體核酸抽取與純化 43
3.5.4.2 聚合酵素鏈連鎖反應（polymerase chain reaction, PCR） 43
3.5.4.3. 序列比對分析 45
3.5.4.4 變性梯度膠電泳 45
3.5.5. 統計方法 45
第四章、結果 47
第一節 新O3：K6型腸炎弧菌對乳鼠致死能力之分析 47
第二節 新O3:K6型腸炎弧菌對乳鼠腸腫脹之分析 47
第三節 新O3:K6型腸炎弧菌於乳鼠腸道生長情形之分析 48
第四節 LPS毒性分析 48
第五節 LPS pattern分析 49
第六節 蛋白質圖譜分析 49
第七節 toxR 序列分析 49
第八節 laf 序列分析 50
第九節 DGGE分析 51
第五章 討論 52
第一節 新O3:K6型腸炎弧菌的毒性 52
第二節 蛋白質圖譜分析 54
第三節 遺傳變異 54
第四節 總結 56
第六章 參考文獻 57

第六章 參考文獻
毛傳忠. 1996. 腸炎弧菌擬鐵調節基因之選殖與特性研究. 東吳大學微生物系碩士論文.台北.
陳育志. 1990. 腸炎弧菌因環境壓力表現在外部之蛋白質分析. 東吳大學微生物系碩士論文.台北.
戴金華. 1986. 鐵離子對腸炎弧菌致病性的影響. 東吳大學微生物系
碩士論文.台北.
Anand, C. P., D. Kakimoto, and T. Sakata. 1981. Growth and antibiotic susceptibility studies on Vibrio parahaemolyticus. Bull. Jpn. Soc. Sci. Fish. 47: 539-545.
Baba, K., H. Shirai, A. Terai, Y. Takeda, and M. Nishibuchi. 1991. Analysis of the tdh gene cloned from a tdh gene- and trh gene-positive strains of Vibrio parahaemolyticus. Microbiol. Immunol. 35:253-8.
Beucha, L. R. 1975. Environmental factors affecting survival and growth of Vibrio parahaemolyticus. A review. J. Milk Food Techn. 38: 476-480.
Bhuiyan, N. A., M. Ansaruzzaman, M. Kamruzzaman, K. Alam, N. R. Chowdhury, M. Nishibuchi, S. M. Faruque, D. A. Sack, Y. Takeda, and G. B. Nair. 2002. Prevalence of the pandemic genotype of Vibrio parahaemolyticus in Dhaka, Bangladesh, and significance of its distribution across different serotypes. J. Clin. Microbiol. 40: 284-286.
Brown, D. F., P. L. Spaulding, and R. M. Twedt. 1977. Enteropathogenicity of Vibrio parahaemolyticus in the ligated rabbit ileum. Appl. Environ. Microbiol. 33:10-14.
Chang, B., H. Taniguchi, H. Miyamoto, and S. Yoshida. 1998. Filamentous bacteriophages of Vibrio parahaemolyticus as a possible clue to genetic transmission. J. Bacteriol. 180: 5094-5101.
Chang, B., S. Yoshida, H. Miyamoto, M. Ogawa, K. Horikawa, M. Nishibuchi, and H. Taniguchi. 2000. A unique and common restriction fragment pattern of the nucleotide sequences homologous to the genome of Vf33, a filamentous bacteriophage, in pandemic strains of Vibrio parahaemolyticus O3:K6 O4:K68, and O1:K untypeable. FEMS Microbiol. Lett. 192: 231-236.
Carruthers, M. M. 1975. Cytotoxicity of Vibrio parahaemolyticus in HeLa cell culture. J. Infect. Dis. 132: 555-560.
Chiou, C. S., S. Y. Hsu, S. I. Chiu, T. K. Wang, and C. S. Chao. 2000. Vibrio parahaemolyticus serovar O3:K6 as cause of unusually high incidence of food-borne disease outbreaks in Taiwan from 1996 to 1999. J. Clin. Microbiol. 38: 4621-4625.
Chowdhury, N. R., S. Chakraborty, T. Ramamurthy, M. Nishibuchi, S. Yamasaki, Y. Takeda, and G. B. Nair. 2000. Molecular evidence of clonal Vibrio parahaemolyticus pandemic strains. Emerg. Infect. Dis. 6: 631-636.
Dai, J. W., Y. S. Lee, and H. C. Wong. 1992. Effects of iron limitation of production of a siderophore, outer-membrane proteins, and hemolysin and on hydrophobicity, cell adherence, and lethality for mice of Vibrio parahaemolyticus. Infect. Immun. 60: 2952-2956.
Dascher, C. C., K. S. Poddar, and J. Maniloff. 1990. Heat shock response in mycoplasmas, genome-limited organisms. J. Bacteriol. 172:1823-1827.
DePaola, A., C. A. Kaysner, J. Bowers, and D. W. Cook. 2000. Environmental investigations of Vibrio parahaemolyticus in Oysters after outbreaks in Washington, Texas, and New York (1997 and 1998). Appl. Environ. Microbiol. 66: 4649-4654.
Dooley, J. S. G., R. Lallier, D. H. Shaw, and T. J. Trust. 1985. Electrophoretic and immunochemical analysis of the lipopolysaccharides from various strains of Aeromonas hydrophila. J. Bacteriol. 164:263-269.
Gingras, S. P., and L. V. Howard. 1980. Adherence of Vibrio parahaemolyticus to human epithelial cell lines. Appl. Environ. Microbiol. 39:369-371.
Hackney, C. R., E. G. Kleeman, B. Ray, and M. L. Speck. 1980. Adherence as a method for differentiating virulent and avirulent strains of Vibrio parahaemolyticus. Appl. Environ. Microbiol. 40:652-658.
Han, T. J., and T. J. Chai. 1992. Electrophoretic and chemical characterization of lipopolysaccharides of Vibrio parahaemolyticus. J. Bacteriol. 174:3140-3146.
Hasegawa, J., Y. Hara-Kudo, T. Nishina, H. Konuma, and S. Kumagai. 2002. Survival of Vibrio parahaemolyticus serovar O3:K6 strains under acidic conditions. Shokuhin Eiseigaku Zasshi 43: 90-94.
Honda, T., Y. Ni, and T. Miwatani. 1988. Purification and characterization of a hemolysin produced by clinical isolate of Kanagawa phenomenon-negative Vibrio parahaemolyticus and related to the thermostable direct hemolysin. Infect. Immun. 56：961-965.
Honda, T., Y. Ni, A. Hata, M. Yoh, and T. Miwatani. 1990. Properties of a hemolysin related to the thermostable direct hemolysin produced by a Kanagawa phenomenon-negative, clinical isolates of Vibrio parahaemolyticus. Can J. Microbiol. 36:395-399.
Honda, T., M. A. Abad-Lapuebla, Y. Ni, K. Yamamoto, and T. Miwatani. 1991. Characterization of a new thermostable direct haemolysin produced by a Kanagawa-phenomenon-negative clinical isolate of Vibrio parahaemolyticus. J. Gen. Microbiol. 137:253-259.
Iguchi, T., S. Kondo, and K. Hisatsune. 1995. Vibrio parahaemolyticus O serotypes from O1 to O13 all produce R-type lipopolysaccharide: SDS-PAGE and compositional sugar analysis. FEMS Microbiol. Lett. 130：287-292.
Iida, T., A. Hattori, K. Tagomori, H. Nasu, R. Naim, and T. Honda. 2001. Filamentous phage associated with recent pandemic strains of Vibrio parahaemolyticus. Emerg. Infect. Dis. 7: 477-478.
Jay, J. M. 1980. Modern food microbiology. D. Van Nostrand Co. New York.
Johnson, D. E., L. Weinberg, J. Cisrkowski, P. West, and R. A. Colwell. 1984 Wound infection caused by Kanagawa-negative Vibrio parahaemolyticus. J. Clin. Microbiol. 20:811-812.
Kaneko, T., and R. R. Colwell. 1975. Adsorption of Vibrio parahaemolyticus onto chitin and copepods. Appl. Microbiol. 29:269-274.
Kaper, J. B., R. K. Campen, R. J. Seidler, M. M. Baldini, and S. Falkow. 1984. Cloning of the thermostable direct or Kanagawa phenomenon-associated hemolysin of Vibrio parahaemolyticus. Infect. Immun. 45:290-292.
Kaysner, C. A., C. Abeyta, P.A. Trost, J. H. Wetherington, K. C. Jinneman, W. E. Hill., and M. M. Wekell. 1994. Urea hydrolysis can predict the potential pathogenicity of Vibrio parahaemolyticus strains isolated in the Pacific Northwest. Appl. Environ. Microbiol. 60：3020-3022.
Kelly, M. T., and E. M. D. Stroh. 1989. Urease-positive, Kamagawa-negetive Vibrio parahaemolyticus from patients and the environment in the Pacific Northwest. J. Clin. Microbiol. 27:2820-2822.
Khan, A. A., S. McCarthy, R. F. Wang, and C. E. Cerniglia. 2002. Characterization of United States outbreak isolates of Vibrio parahaemolyticus using enterobacterial repetitive intergenic consensus (ERIC) PCR and development of a rapid PCR method for detection of O3:K6 isolates. FEMS Microbiol. Lett. 206: 209-214.
Koga, T., and T. Kawata. 1991. Comparative characterization of inducible and virulent Vibrio parahaemolyticus bacteriophages having unique head projections. Microbiol. Immunol. 35: 49-58.
Koga, T., and K. Takumi. 1995. Nutrient starvation induces cross protection against heat, osmotic, or H2O2 challenge in Vibrio parahaemolyticus. Microbiol. Immunol. 39:213-215.
Lee, C. Y., S. H. Pan, and C. H. Chen. 1995. Sequence of a cloned pR72H fragment and its use for detection of Vibrio parahaemolyticus in shellfish with the PCR. Appl. Environ. Microbiol. 61:1311-1317.
Lee, S. E., S. H. Shin, S. Y. Kim, Y. R. Kim, D. H. Shin, S. S. Chung, Z. H. Lee, J. Y. Lee, K. C. Jeong, S. H. Choi and J. H. Rhee. 2000. Vibrio vulnificus has the transmembrane transcription activator ToxRS stimulating the expression of the hemolysin gene vvhA. J. Bacteriol. 182: 3405-3415.
Merrell, B. R., R. I. Walker, and S. W. Joseph. 1984. In vitro and in vivo pathologic effects of Vibrio parahaemolyticus on human epithelial cells. Can. J. Microbiol. 30:381-388.
Matsumoto C., J. Okuda, M. Ishibashi, M. Iwanaga, P. Garg, T. Rammamurthy, H. Wong, A. DePaola, Y. B. Kim, M. J. Albert, and M. Nishibuchi. 2000. Pandemic spread of an O3:K6 clone of Vibrio parahaemolyticus and emergence of related strains evidenced by arbitrarily primed PCR and toxRS sequence analyses. J. Clin. Microbiol. 38: 578-585.
Makino,.K.,.K.Oshima,,.K..Kurokawa,.K..Yokoyama,.T..Uda,.K..Tagomori, Y. .Iijima, M. .Najima,.M. Nakano, A. Yamashita, Y..Kubota, S. Kimura, T. Yasunaga, T. Honda, H. Shinagawa, M. Hattori, and T. Iida. 2003. Genome sequence of Vibrio parahaemolyticus: a pathogenic mechanism distinct from that of V . cholerae. Lancet 361:743-749
Morrison, D. C., and R. J. Ulevitch. 1978. The effect of bacterial endotoxins on host mediation systems. Am. J. Pathol. 93:527-617.
Muntada-Garriga, J. M., J. J. Rodriguez-Jerez, E. I. Lopez-Sabater, and M. T. Mora-Ventura. 1995. Effect of chill and freezing temperatures on survival of Vibrio parahaemolyticus inoculated in homogenates of oyster meat. Lett. Appl. Microbiol. 20:225-227.
Myer, R. M., Maniatis, T., and L. S. Lerman. 1987. Detection and localization of single base pair changes by denaturing gradient gel eletrophoresis. Meth. Enzymol. 155:501-512
Nagayama, K., T. Oguchi, M. Arita, and T. Honda. 1994. Correlation between cell-associated mannose-sensitive hemagglutination by Vibrio parahaemolyticus and adherence to a human colonic cell line Caco-2. FEMS Microbiol. Lett. 120:207-210.
Nagayama, K., K. Yamamoto, T. Mitawani, and T. Honda. 1995. Characterisation of a hemolysin related to VP-TDH produced by a Kanagawa phenomenon-negative clinical isolate of Vibrio parahaemolyticus. J. Med. Microbiol. 42:83-90.
Nakasone, N., and M. Iwanaga. 1990. Pilli of a Vibrio parahaemolyticus strain as a possible colonization factor. Infect. Immun. 56:61-69.
Nakasone N, M. Ikema, N. Higa, T. Yamashiro, and M. Iwanaga. 1999. A Filamentous Phage of Vibrio parahaemolyticus O3:K6 Isolated in Laos. Microbiol. Immunol. 43: 385-388.
Nakasone, N., S. Insisengmay, and M. Iwanaga. 2000. Characterization of the pili isolated from Vibrio parahaemolyticus O3:K6. Southeast Asian J. Trop. Med. Public Health 31: 360-365.
Nasu, H., T. Iida, T. Sugahara, Y. Yamaichi, K. S. Park, K. Yokoyama, K. Makino, H. Shinagawa, and T. Honda. 2000. A filamentous phage associated with recent pandemic Vibrio parahaemolyticus O3:K6 strains. J. Clin. Microbiol. 38: 2156-2161.
Nishibuchi, M., A. Fasano, R. G. Russell, and J. B. Kaper. 1992. Enterotoxigenicity of Vibrio parahaemolyticus with and without genes encoding thermostable direct hemolysin. Infect. Immun. 60:3539-3545.
Nishibuchi, M., and J. B. Kaper. 1990. Duplication and variation of the thermostable direct hemolysin (tdh) gene in Vibrio parahaemolyticus. Mol. Microbiol. 4:87-99.
Nishibuchi, M., and J. B. Kaper. 1995. Thermostable direct hemolysin gene of Vibrio parahaemolyticuss: a virulence gene acquired by a marine bacterium. Infect. Immun. 63:2093-2099.
Nishibuchi, M., K. Kumagai, and J. B. Kaper. 1991. Constribution of the tdh1 gene of Kanagawa phenomenon-positive Vibrio parahaemolyticus to production of extracellular thermostable direct hemolysin. Microb. Pathog. 11:453-460.
Nishibuchi, M., T. Taniguichi, T. Misawa, V. Khaeomanee-Iam, T. Honda, and T. Miwatani. 1989. Cloning and nucleotide sequence of the gene (trh) encoding the hemolysin related to the thermostable direct hemolysin of Vibrio parahaemolyticus. Infect. Immun. 57:2691-2697.
O’Brien, A. D., M. E. Chen, R. K. Holmes, J. Kaper, M. M. Levine.
1984. Environmental and human isolates of Vibrio parahaemolyticus produce a Shigella dysenteriae 1 (Shiga)-like cytotoxin. Lancet 1:77-78.
Okuda J., M. Ishibashi, E. Hayakawa, O. T. Nishin, A. Y. Taked, A. K. Mukhopadhyay, S. Garg, S. K. Bhattacharya, G. B. Nair, and M. Nishibuchi. 1997. Emergence of a unique O3:K6 clone of Vibrio parahaemolyticus in Calcutta, India, and isolation of strains from the same clonal group from Southeast Asian travelers arriving in Japan. J. Clin. Microbiol. 35: 3150-3155.
Provenzano, D., and K. E. Klose. 2000. Altered expression of the ToxR-regulated porins OmpU and OmpT diminishes Vibrio cholerae bile resistance, virulence factor expression, and intestinal colonization. Proc. Natl. Acad. Sci. U. S. A. 97:10220-4.
Reyes, A. L., R. G. Crawford, P. L. Spaulding, J. T. Peeler, and R. M. Twedt. 1983. Hemagglutination and adhesiveness of epidemiologically distinct strains of Vibrio parahaemolyticus. Infect. Immun. 39:721-725
Sakazaki, R., K. Tamura, T. Kato, Y. Kato, Y. Obary, S. Yamai, and K. Hobo. 1968. Studies on the enteropathogenic, facultatively halophilic bacteria, Vibrio parahaemolyticus. III. Enteropathogenicity. Jpn. J. Med. 21:313-24
Sanyal, S. C., and P. C. Sen. 1974. Human volunteer study on the pathogenicity of Vibrio parahaemolyticus, p.227-235. In T. Fujino, G. Sakaguchi, R.Sslazaki, and Y.Takeda. (ed.), International symposium on Vibrio parahaemolyticus. Saiko Publ. Co., Tokyo.
Sarkar, B. L., R. Kumar, S. P. De, and S. C. Pal. 1987a. Hemolytic activity of the lethal toxin production by environmental strains of Vibrio parahaemolyticus. Appl. Environ. Microbiol. 53:2696-2698.
Sarkar, B. L., R. Kumar, S. P. De, and S. C. Pal. 1987b. Observation on a 65-Kilodalton protein isolated from Kanagawa positive strains of Vibrio parahaemolyticus. Can. J. Microbiol. 33:1113-1116.
Suthienkul, O., M. Ishibashi, T. Iida, N. Nettip, S. Supavej, B. Eampokalap, M. Makino, and T. Honda. 1995. Urease production correlates with possession of the trh gene in Vibrio Parahaemolyticus strain s isolated in Thailand. J. Infect. Dis. 172:1405-1408.
Takeda, Y. 1983. Thermostable direct hemolysin of Vibrio parahaemolyticus. Pharmac. Ther. 19:123-146.
Taniguchi H, K. Sato, M. Ogawa, T. Udou, and Y. Mizuguchi. 1984. Isolation and characterization of a filamentous phage, Vf33, specific for Vibrio parahaemolyticus. Microbiol. Immunol. 28: 327-337.
Wong, H. C., C.C. Liu, C. M. Yu, and Y. S. Lee. 1996. Utilization of iron sources and its possible roles in the pathogenesis of Vibrio parahaemolyticus. Microbiol.Immunol. 40:791-798.
Wong, H. C., K. T. Lu, T. M. Pan, C. L. Lee, and D. Y. C. Shih. 1996. Subspecies typing of Vibrio parahaemolyticus by pulsed-field gel electrophoresis. J.Clin.Microbiol. 34:1535-1539.
Wong, H. C., P. Y. Peng, J. M. Han, C. Y. Chang, and S. L. Lan. 1998. Effect of mild acid treatment on the survival, enterotoxigenicity and protein production in Vibrio parahaemolyticus. Infect.Immun. 66:3066-3071.
Wong, H. C., S. H. Liu, L. W. Ku, I. Y. Lee, T. K. Wang, Y. S. Lee, C. L. Lee, L. P. Kuo, and D. Y. C. Shih. 2000a. Characterization of Vibrio parahaemolyticus isolates obtained from foodborne illness outbreaks during 1992 through 1995 in Taiwan. J. Food Prot. 63:900-906.
Wong, H. C., S. H. Liu, T. K. Wang, C. L. Lee, C. S. Chiou, D. P. Liu, M. Nishibuchi, and B. K. Lee. 2000b. Characteristics of Vibrio parahaemolyticus O3:K6 from Asia. Appl. Environ. Microbiol. 66: 3981-3986.
Yamamoto, K., and T. Yokota. 1989. Adherence targets of Vibrio parahaemolyticus in human small intestines. Infect. Immun. 57: 2410-2419.
Yeung, P. S., M. C. Hayes, A. DePaola, C. A. Kaysner, L. Kornstein, and K. J. Boor. 2002. Comparative phenotypic, molecular, and virulence characterization of Vibrio parahaemolyticus O3:K6 isolates. Appl. Environ. Microbiol. 68: 2901-2909.
Yoh, M., T. Honda, and T. Miwatani. 1986. Purificcation and partial characterization of non-O1 Vibrio cholerae hemolysin that cross-reacts with thermostable direct hemolysin of Vibrio parahaemolyticus. Infect. Immun. 52:319-322.
Yung, B. K., J. Okuda, C. Matsumoto, N. Takahashi, S. Hashimoto, and M. Nishibuchi. 1999. Identification of Vibrio parahaemolyticus Strains at the Species Level by PCR Tarteted to the toxR Gene. J. Clin. Microbiol. 37:1173-1177