臺灣博碩士論文加值系統

本實驗將香魚分離出之親水性產氣單胞菌Aeromonas hydrophila，毒力因子aerolysin之前軀物preproaerolysin之基因加以選殖與表現。得到preproaerolysin基因完整的序列，含有1461個核酸之DNA片段，在接入表現載體後，可以大量表現出487個氨基酸，分子量約為59kDa的基因重組preproaerolysin蛋白。由基因選殖所製備出來的preproaerolysin，經trypsin活化以後，亦保留本毒力因子之功能，會對blood agar產生溶血的現象，且對於Vero細胞株有細胞溶解的毒性，但基因重組 aerolysin 在46℃加熱5分鐘後就失去溶血活性。在動物試驗中，會造成小白鼠的死亡，在兔迴腸試驗中，會造成兔子迴腸的出血水腫。

The preproaerolysin of Aeromonas hydrophila isolated from ayu was cloned and expressed. The complete preproaerolysin gene was 1461 base pairs, which contained 487 amino acids, and the molecular weight was about 59kDa. The recombinant preproaerolysin retained its toxicity after activated with trypsin, and the property was deactivated by treating at 46℃ for 5 min. Hemolytic ability on blood agar and cytolytic ability on Vero cell was noted in vitro. It caused the death of mice and ileum edema with hemorrhage in the rabbit ileum loop test in vivo.

目錄
目錄 I
圖表次 III
中文摘要 IV
英文摘要 V
第一章 緒言 1
第二章 文獻探討 2
第一節 Aeromonas hydrophila之生物分類學上的特徵 2
第二節 環境與A. hydrophila之關係 3
2-1 溫度 3
2-2 鹽度適應性 4
2-3 在各種培養基的生產毒素能力 4
2-4 對pH值之適應性 5
2-5 抗藥性 5
2-6 環境適應性 6
第三節 A. hydrophila 產生之病害 7
3-1 對魚類的致病性 7
3-2 對人類的致病性 9
3-3 A. hydrophila 的生物防治 10
第四節 A. hydrophila 之毒力因子 10
4-1 外毒素 11
4-1-1 磷脂脢 12
4-1-2 溶血素 13
4-1-3 腸毒素 15
4-1-4 細胞毒素 16
4-1-5 蛋白脢 16
4-1-6 其他細菌外毒素 17
4-2 內毒素 18
4-3 Aerolysin 19
第五節 以大腸菌大量生產特定蛋白的重要性 23
第三章 材料與方法 24
3-1 菌株 24
3-2 質體 24
3-3 培養基 25
3-4 藥品 26
3-5 實驗方法 30
3-5-1 細菌核酸之萃取 30
3-5-2 PCR 反應增幅核酸片段 30
3-5-3 PCR產物的分離與回收 31
3-5-4 小規模質體製備 31
3-5-5 限制脢的作用 33
3-5-6 接合反應 34
3-5-7 勝任細胞(competent cells)的製作 34
3-5-8 細胞轉型反應 35
3-5-9 核酸之定序 36
3-5-10 核酸之分析方法 36
3-5-11 細菌蛋白質的誘導產生 37
3-5-12 SDS-PAGE電泳 37
3-5-13 基因重組preproaerolysin的大量回收 40
3-5-14 基因重組preproaerolysin的refolding 41
3-5-15 基因重組preproaerolysin的蛋白質定量 41
3-5-16 小白鼠多株抗體血清(Mouse polyclonal antisera) 42
3-5-17 多株抗體血清之吸附反應 42
3-5-18 西方墨點法 43
3-5-19 動物血液收集與血液培養基之製作 45
3-5-20 基因重組aerolysin對各種動物血之溶血性狀測試 46
3-5-21 Vero 細胞生物毒性試驗 47
3-5-22 溫度對於重組蛋白毒性之影響 47
3-5-23 小白鼠毒性試驗 48
3-5-24 兔子迴腸結紮試驗 49
第四章 結果 50
4-1 以PCR增幅preproaerolysin基因之電泳 50
4-2 Preproaerolysin 基因定序與與轉譯氨基酸之結果 51
4-3 基因重組preproaerolysin製備之結果 53
4-4 基因重組preproaerolysin之免疫學確認 55
4-5 基因重組aerolysin溶血性狀之測試 56
4-6 不同濃度與溫度下基因重組aerolysin對Vero cell之細胞毒性 58
4-7 基因重組 aerolysin對於小白鼠之致病性 60
4-8 基因重組 aerolysin對於兔腸之腸毒性 60
第五章 討論 62
5-1 Preproerolysin 基因之選殖 62
5-2 Preproaerolysin 基因之定序 65
5-3 基因重組preproaerolysin的大量製備、回收與免疫學之確認 66
5-4 基因重組aerolysin 生物活性之測試 68
5-4-1 基因重組aerolysin溶血性狀之測試 68
5-4-2 基因重組 aerolysin 對Vero cell之細胞毒性 69
5-4-3 溫度對於基因重組 aerolysine之影響 69
5-4-4 基因重組 aerolysin對於小白鼠之致病性 70
5-4-5 基因重組 aerolysin對於兔腸之腸毒性 70
參考文獻 72
圖表次
圖1、pET29a 表現載體 25
圖2、實驗設計流程圖 27
圖3、以PCR增幅A. hydrophila 台灣分離株之
preproaerolysin基因之DNA片段 50
圖4、A. hydrophila台灣分離株preproaerolysin
基因之核酸與氨基酸序列 51
圖5、以SDS-PAGE電泳分析三株不同品系E. coli以IPTG
誘導合成基因重組preproaerolysin的產量與分子量 54
圖6、以Western blot分析小白鼠抗A. hydrophila
之多株抗體與基因重組 preproaerolysin之特異性結合 55
圖7、基因重組aerolysin對各種動物之血液培養基之溶血性測試 57
圖8、基因重組 aerolysin在各種不同條件之下對Vero cell之影響 59
圖9、腹腔注射基因重組 aerolysin 310.9μg/mouse後死亡之老鼠解剖61
圖10、基因重組 aerolysin在兔迴腸結紮試驗 61
圖11、基因重組表現載體 pET29aer 63
表一、A. hydrophila可能的外泌性致病因子 11

參考文獻：
1.宋延齡，陳秀男，郭光雄。 Aeromonas hydrophila (A. liquetaciens)免疫鰻魚(Anguilla japonica)後之凝集抗體與保護作用。 臺灣水產學會刊4: 25-49， 1976.
2.周正俊，蔡漢蓉。 不同pH與溫度下亞硝酸鈉對Aeromonas hydrophila之抗菌活性。 食品科學23: 159-167，1996。
3.林敏雄，陳秀男，郭光雄。 臺灣地區魚病研究之回顧。 農發會漁業特刊第九號，魚病研究專集(五)： 1-9，1983。
4.林曜松，蕭世民。 臺灣魚池生態環境與魚病關係之研究(I)臺灣鰻魚疾病之統計分析。 魚病研究專集(一)： 57-61，1977。
5.吳慈佩。自台灣養殖香魚分離之親水性產氣單胞菌的表現型和基因型分析及病原性試驗。 國立台灣大學獸醫學研究所碩士論文， 台北， 1-65， 1998。
6.施養志，賴紹伶，朱淑儀，王貞懿。 以回歸曲面法探討溫度、pH值和氯化鈉含量對 Aeromonas hydrophila DNA group1生長之影響。 中華生質能源會會誌14： 85-93，1995。
7.郭光雄，鐘虎雲。 養鰻池之R+抗藥細菌。 農委會漁業特刊第十號，魚病研究論文集(一)細菌及免疫專集： 70-77，1987。
8.陳林楊，簡秋源。 台灣養殖黑鯛爛尾病病原菌Aeromonas hydrophila之分離。 農委會漁業特刊第十號，魚病研究論文集(一)細菌及免疫專集： 78-91，1987。
9.劉昭君，林麗英，莊榮豐，張南驥，蔡文城，胡小婷。 臨床Aeromonas hydrophila菌株溶血素基因之選殖與核酸定序。 中華微免雜誌23： 134-146，1990。
10.劉富光。 臺灣養殖泥鰍之疾病。 中國水產324： 14-18，1979。
11.劉朝鑫，郭宗甫。 魚類病原菌抗藥性之研究-I，養鰻地Aeromonas hydrophila之抗藥性。 農委會漁業特刊第四號，魚病研究專集(七)： 169-175，1985。
12.劉昭君，林麗英，莊榮豐，張南驥，蔡文城，胡小婷。 臨床A. hydrophila 菌株溶血素基因之選殖及核酸定序。 中華微免雜誌 23：134-146，1990。
13.賴信志，胡小婷，劉世東，蔡文城。 Aeromonas hydrophila臨床菌株具有溶血素基因及與弧菌毒素基因相同核酸序列之片段。 中華微免雜誌23： 126-133，1990。
14.蕭世民，陳孝禹。 臺灣地區牛蛙(Rana catesbiana)、淡水長腳大蝦(Marrobrachium rosenbergii)及淡水養殖魚類發現細菌及寄生蟲病。 魚病研究專集(一)： 19-21，1977。
15.鍾虎雲，郭光雄。 養殖牛蛙細菌相之研究-II。 農委會漁業特刊第八號，魚病研究專集(八)： 1-5，1986。
16.Aaron B, Ingham MP. A lipase of Aeromonas hydrophila showing nonhemolytic phospholipase C activity. Cur Microbiol 31: 28-33, 1995.
17.Abeyta CA, Wekell MM, Sullivn JJ, Stelma GN. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. J. Food Prot. 49: 643-646, 1986.
18.Abrami L, Fivaz M, Decroly E, Seidah NG, Jean F, Thomas G, Leppla SH, Buckley JT, van der Goot FG. The pore-forming toxin proaerolysin is activated by furin. J Biol Chem.273: 32656-32661, 1998.
19.Abrami L, Fivaz M, Glauser PE, Parton RG, van der Goot FG. A pore-forming toxin interacts with a GPI-anchored protein and causes vacuolation of the endoplasmic reticulum. J Cell Biol 140: 525-40, 1998.
20.Allan. BJ, Stevenson. RMW. Extracellular virulence factors of Aeromonas hydrophilain fish infections. Can. J. Microbiol 27: 1114-1122, 1981.
21.Amin NE, Abdallah IS, Elallawy T, Ahmed SM. Motile Aeromonas septicaemia among Tilapia nilotica (Sarotherodon niloticus) in upper Egypt. Fish Pathology 20: 93-97, 1985.
22.Aoki T, Hirono I. Cloning and characterization of the haemolysin determinants from Aeromonas hydrophila. J Fish Dis. 14: 303-312, 1991.
23.Aoki T, Holland BI. The outer membrane proteins of fish pathogens Aeromonas hydrophila, Aeromonas salmonicida and Edwardsiella tarda. FEMS Microbiol Lett. 27: 299-305, 1985.
24.Asao T, Kozaki S, Kato K, Kinoshita Y, Otsu K, Uemura T, Sakaguchi G. Purification and characterization of an Aeromonas hydrophila hemolysin. J. Clin. Microbiol. 24: 228-232, 1986.
25.Asao T, Kozaki S, Kato K, Kinoshita Y, Uemura T, Sakaguchi G. Purification and some properties of Aeromonas hydrophila hemolysin. Infect Immun. 46: 122-127, 1984.
26.Ascencio F, Ljungh A, Wadstrom T. New lectins and other putative adhesins in Aeromonas hydrophila. Experientia 47: 414-416, 1991.
27.Austin B, Austin DA. Disease in Farmed and Wild Fish. Ellis Horwood Ltd, Chichester. 1987.
28.Barer MR, Milliership SE, Tabaqchali S. Relationship of toxin production to species in the genus Aeromonas. J Med Microbiol. 22: 303-309, 1986.
29.Becker GW, Hsiung HM. Expression, secretion and folding of human growth hormone in Escherichia coli. FEBS 204: 145-150, 1986.
30.Benson SA, Hall MN, Silhavy TJ. Genetic analysis of protein export in Escherichia coli K12. Ann Rev Biochem. 54: 101-134, 1985.
31.Bernheimer AW, Avigad LS, Avigad G. Interactions between aerolysin, erythrocytes, and erythrocyte membranes. Infect Immun.11: 1312-1319, 1975.
32.Bernheimer AW, Avigad LS. Partial characterization of aerolysin, a lytic exotoxin from Aeromonas hydrophila. Infect Immun. 9: 1016-1021, 1974.
33.Betly MJ, Miller VL, Mekalanos JJ. Genetics of bacterial enterotoxins. Ann Rev Microbiol. 40: 577-605, 1986.
34.Bhat P, Shanthakumari S, Rajan D. The characterization and significance of lesiomonas shigelloides and Aeromonas hydrophila isolated from an epidemic of diarrhoea. Indian J Med Res 62: 1051-1060, 1974.
35.Bloch S, Monteil H. Purification and characterization of Aeromonas hydrophila beta-hemolysin. Toxicon 27: 1279-1287, 1989.
36.Boulanger Y, Lallier R, Cousineau G. Isolation of enterotoxigenic Aeromonas from fish. Can J Microbiol. 23: 1161-1164, 1977.
37.Brenden R, Janda JM. Detection, quantitaion and stability of β-haemolysin of Aeromonas spp. J Med Microbiol. 24: 247-251, 1987.
38.Buchanan RL. The "New" pathogens: An update of selected examples. Assoc. Food & Drug Officials. Quarterly Bulletin. 48: 142-155, 1984.
39.Buchanan RL, Palumbo SA. Aeromonas hydrophila and Aeromonas sobria as potential food poisoning species: a review. J Food Saf. 7: 15-29, 1985.
40.Buckley JT, Halasa LN, Luns KD, MacIntyre S. Purification and some properties of the hemolytic toxin aerolysin. Can J Biochem. 59: 430-436, 1981.
41.Buckley JT, Halasa LN, MacIntyre S. Purification and partial characterization of a bacterial phospholipid-cholesterol acyltransferase. J Biol Chem. 257: 3320-3325, 1982.
42.Buckley JT, Howard SP. Aerolysin from Aeromonas hydrophila in: Microbial Toxins. Methods Enzymol. 165: 193-200, 1988.
43.Burke V, Gracey M, Robinson J, Beaman J, Bundell C. The microbiology of childhood gastroenteritis: Aeromonas species and other infective agents. J Infect Dis. 148: 68-74, 1983.
44.Burke V, Robinson J, Beny RJ, Gracey M. Detection of enterotoxins of Aeromonas hydrophilaby a suckling-mouse test. J Med Microbiol. 14: 401-408, 1981.
45.Cabiaux V, Buckley JT, Wattiez R, Ruysschaet JM, Parker MW, van der Goot, FG. Conformational changes in aerolysin during the transition from the water-soluble protoxin to the membrane channel. Biochemistry 36: 15224-15232, 1997.
46.Chabot DJ, Thune RL. Proteases of the Aeromonas hydrophila complex: identification, characterization and relation to virulence in channel catfish, Ictalurus punctatus (Rafinesque). J Fish Dis. 14: 171-183, 1991.
47.Chakraborty T, Huhle B, Bergbaure H, Goebel W. Cloning, expression, and mapping of the Aeromonas hydrophila aerolysin gene determinant in Escherichia coli K-12. J. Bacteriol. 167: 368-374, 1986.
48.Chakraborty T, Montenegro MA, Sanyal SC, Helmuth R, Bulling E, Timmis KN. Cloning of enterotoxin gene from Aeromonas hydrophila provides conclusive evidence of production of a cytotonic enterotoxin. Infect. Immun. 46: 435-441, 1984.
49.Chakraborty T, Schmid A, Notermans S, Benz R. Aerolysin of Aeromonas sobria: evidence for formation of ion-permeable channels and comparison with alpha-toxin of Staphylococcus aureus. Infect Immun 58: 2127-2132, 1990.
50.Chang TM, Liu CC, Chang MC. Cloning and sequence analysis of the gene (eprA1) encoding an extracellular protease from Aeromonas hydrophila. Gene. 199: 225-229, 1997.
51.Chopra AK, Houston CW. Purification and partial characterization of a cytotonic enterotoxin produced by Aeromonas hydrophila. Can J Microbiol. 35: 719-727, 1989.
52.Chopra AK, Houston CW, Genaux CT, Dixon JD, Kurosky A. Evidence for production of an enterotoxin and cholera toxin crossreactive factor by Aeromonas hydrophila. J Clin Microbiol. 24: 661-664, 1986.
53.Chung KT, Yu FP. Survival of Aeromonas hydrophila and Escherichia coli in aquatic environments. Chinese J Microbiol Immunol. 23: 181-188, 1990.
54.Colwell RR, MacDonell MR, DeLey J. Proposal to recognize the family Aeromonadaceae fam. Int J Syst Bacteriol. 36: 473-477, 1986.
55.Condon SG, Garcia ML, Otero A, Sala FJ. Effect of culture age, pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila. J. Appl. Bacteriol. 72: 322-326, 1992.
56.Cordingley FT, Rajanayagam A. Aeromonas hydrophila bacteremia in haematological patients. Med. J. Aust. 1: 364-365, 1981.
57.Chuang YC, Chiou SF, Su JH, Wu ML, Chang MC. Molecular analysis and expression of the extracellular lipase of Aeromonas hydrophila MCC-2. Microbiol. 143: 803-812, 1997.
58.Delepelaire P, Wandersman C. Protein secretion in gram-negative bacteria. J Biol Chem. 265: 17118-17125, 1990.
59.De SN, Chaterje DN. An experimental study of the mechanism of action of Vibrio cholerae on the instinal mucous membrane. J Pathol Bacteriol. 66:559-562, 1953.
60.Donta ST, Haddow AD. Cytotoxic activity of Aeromonas hydrophila. Infect. Immun. 21:989-993, 1978.
61.Dooley JS, Trust TJ. Surface protein composition of Aeromonas hydrophila strains virulent for fish: Identification of a surface array protein. J Bacteriol. 170: 499-506, 1988.
62.Dooley JS, Lallier GR, Shaw DH, Trust TJ. Electrophoretic and immunochemical analyses of the lipopolysaccharides from Aeromonas hydrophila. J Bacteriol. 164: 263-269, 1985.
63.Dryden M, Munro R. Aeromonas septicemia: relationship of species and clinical features. Pathology, 21: 111-114, 1989.
64.Eley A, Geary I, Wilcox MH. Growth of Aeromonas spp. at 4℃ and related toxin production. Lett. Appl. Microbiol. 16: 36-39, 1993.
65.Ellis AE. An appraisal of the extracellular toxin of Aeromonas salmonisida ssp. J. Fish Dis., 14: 265-277, 1991.
66.Filloux A, Bally M, Ball G, Akrim M, Tommassen J, Lazdunski A. Protein secretion in gram-negative bacteria transport across the outer membrane involves common mechanisms in different bacteria. EMBO J. 9: 4323-4329, 1990.
67.Freij BJ. Aeromonas: Biology of the organism and disease in children. Pediatr Infect Dis J 3: 164-175, 1984.
68.Garland WJ, Buckley JT. The cytolytic toxin aerolysin must aggregate to disrupt erythrocytes, and aggregation is stimulated by human glycophorin. Infect Immun. 56:1249-1253, 1988.
69.Gobius KS, Pemberton JM. Molecular cloning, characterization, and nucleotide sequence of an extracellular amylase gene from Aeromonas hydrophila. J Bacteriol. 170: 1325-1332, 1988.
70.Goguen JD, Hoe NP, Subrahmanyam YVBK. Protease and bacterial virulence: a view from the trenches. Infect Agents Dis. 4: 47-54, 1995.
71.Green MJ, ,Bucley JT. Site-directed mutagenesis of the hole-forming toxin aerolysin. Studies on the roles of histidines in receptor dinding and oligomerizationn of a cytolytic protein toxin. Biochemistry. 29: 2177-2180, 1990.
72.Hanes DE, Chanader DKF. The role of a 40-megadaiton plasmid in the adherence and hemolytic properties of Aeromonas hydrophila. Microb Pathog. 15: 313-317, 1993.
73.Hazen TC, Fliermans CB, Hirsch RP, Esch GW. Prevalence and distribution of Aeromonas hydrophila in the United States. Appl Environ Microbiol. 36: 731-738, 1978.
74.Hirono I, Aoki T. Cloning and characterization of three hemolysin genes from Aeromonas hydrophila. Microb pathog. 15: 269-282, 1993.
75.Hirono I, Aoki T. Nucleotide sequence and expression of an extracellular hemolysin gene of Aeromonas hydrophila. Microb Pathog. 11:189-197, 1991.
76.Hirono I, Aoki T, Asao T, Kozaki S. Nucleotide sequences and characterization of hamolysin genes from Aeromonas hydrophila and Aeromonas sorbia. Microb pathog. 13: 433-446, 1992.
77.Ho ASY, Mietzner TA, Smith AJ, Schoolnik GK. The pili of Aeromonas hydrophila: identification of an environmentally regulated ‘mini pilin’. J Exp Med. 172: 795-806, 1990.
78.Hokama A, Honma Y, Nakasone N. Pili of an Aeromonas strain as a possible colonization factor. Microbiol Immunol. 34: 901-915, 1990.
79.Holmberg SD, Farmer JJ. Aeromonas hydrophilaand Plesiomonas shigelloides as causes of intestinal infections. J. Clin. Microbiol. 633-639, 1984.
80.Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST. Bergey’s manual of determinative bacteriology, 9 th ed., Williams and Wilkins Co, Baltimore, 546-548 1994.
81.Howard SP, Buckley JT. Activation of the hole-formation toxin aerolysin by extracellular processing. J Bacteriol., 163: 336-340, 1985.
82.Howard SP, Buckley JT. Intracellular accumulation of extracellular proteins by pleiotropic export mutants of Aeromonas hydrophila. J Bacteriol. 154: 413-418, 1983.
83.Howard SP, Buckley JT. Membrane glycoprotein receptor and hole-forming properties of a cytolytic goprotein toxin. Biochemistry. 21: 1662-1667, 1982.
84.Howard SP, Buckley JT. Molecular cloning and expression in Escherichia coli of the structural gene for the hemolytic toxin aerolysin from Aeromonas hydrophila. Mol Gen Benet. 204: 289-295, 1986.
85.Howard SP, Buckley JT. Protein export by a gram- negative bacterium: production of aerolysin by Aeromonas hydrophila. J Bacteriol. 161: 1118-1124, 1985.
86.Howard SP, Critch J, Bedi A. Isolation and analysis of eight exe genes and their involvement in extracellular protein secretion and outer membrane assembly in Aeromonas hydrophila. J Bacteriol. 175: 6695-6703, 1993.
87.Howard SP, Garland WJ, Green MJ, Buckley JT. Nucleotide sequence of the gene for the hole-forming toxin aerolysin of Aeromonas hydrophila. J Bacteriol. 169: 2869-2871, 1987.
88.Hudson JA. Variation in growth kinetics and phenotype of Aeromonas spp. from clinical, meat processing and freshfood source. Int J Food Microbiol. 16: 131-139, 1992.
89.Ingham AB, Pemberton J M. A lipase of Aeromonas hydrophila showing nonhemolytic phospholipase C activity. Curr. Microbiol. 31: 28-33, 1995.
90.Janda JM. Biochemical and exoenzymatic properties of Aeromonas species. Diagn Microbiol Infect Dis 3: 223-232, 1985.
91.Janda JM. Recent advances in the study of taxonomy, pathogenicity and infectious syndromes associated with the genus Aeromonas. Clin Microbiol Rev. 4: 397-410, 1991.
92.Janda, JM, Abbott SL. Evolving concepts regarding the genus Aeromonas: an expanding panorama of species, disease presentation and unanswered questions. Clin Infect Dis. 27: 332-344, 1998.
93.Janda JM, Guthertz LS, Kokka RP, Shimada T. Aeromonas species in septicemia: laboratory characteristics and clinical observations. Clin Infect Dis. 19: 77-83, 1994.
94.Johnson WM, Lior H. Cytotoxicity and suckling mouse reactivity of Aeromonas hydrophila isolated from human sources. Can J Microbiol. 27: 1019-1027, 1981.
95.Kanai K, Takagi Y. Alpha-hemolytic toxin of Aeromonas hydrophila produced in vivo. Fish Pathology. 21:245-250, 1986.
96.Kaper JB, Lockman H, Colwell RR, Joseph SW. Aeromonas hydrophila: ecology and toxigenicity of isolates form an estuary. J Appl Bacteriol 50: 359-377, 1981.
97.Keusch GT, Donta ST. Classification of enterotoxins on the basis of activity in cell culture. J Infect Dis. 131: 58-63, 1975.
98.Khardori N, Fainstein V. Aeromonas and Plesiomonas as etiological agent. Ann Rev Microbiol. 42: 395-419, 1988.
99.Kirov SM, Ardestani EK, Hayward LJ. The growth and expression of virulence factors at refrigeration temperature by Aeromonas strains isolated from foods. Int J Food Microbiol. 20: 159-168, 1993.
100.Kirov SM, Hui DS, Hayward LJ. Milk as a potential source of Aeromonas gastrointestinal infection. J Food Prot. 56: 306-312, 1993.
101.Kozaki S, Asao T, Kamata Y, Sakaguchi G. Characterization of Aeromonas sobria hemolysin by use of monoclonal antibodies against Aeromonas hydrophila hemolysins. J Clin Microbiol. 27:1782-1786, 1989.
102.Krovacek K, Faris A, Mknsson I. Growth of and toxin production by Aeromonas hydrophila and Aeromonas sobria at low temperature. Int J Food Microbiol. 13: 165-176, 1991.
103.Kusuda R, Chen C, Kawai K. Changes in the agglutinating antibody titer and serum protein composition of colored carp after immunization with Aeromonas hydrophila. Fish Pathology. 22: 141-146, 1987.
104.Kusuda R, Chen C, Kawai K. The immunoglobulin of colored carp, Cyprinus carpio, against Aeromanas hydrophila. Fish Pathology. 22: 179-183, 1987.
105.Kanai K, Wakabayashi H. Purification and some properties of protease from Aeromonas hydrophila. Bulletin of the Japanese Society of Scientific Fisheries. 50: 1367-1374, 1984.
106.Lamers CH, Muiswinkel WBV. Natural and acquired agglutinins to Aeromonas hydrophila in carp (Cyprinus carpio). J Fish Aquat Sci. 43: 619-624, 1986.
107.Lee SY, Yin Z, Ge R, Sin YM. Isolation and characterization of fish Aeromonas hydrophila adhesins important for in vivo epithelial cell invasion. J Fish Dis 20: 169-175, 1997.
108.Lee LN, Luh KT, Hsieh CW. Bacteremia due to Aeromonas hydrophila: a report of 40 episodes. J Formas Med Assoc. 85: 123-132, 1986.
109.Leung KY, Lim TM, Lam TJ, Sin YM. Morphology changes in carp epithelial cells infected with Aeromonas hydrophila. J Fish Dis 19: 167-174, 1996.
110.Leung KY, Stevenson MW. Characterization and distribution of extracellular proteases from Aeromonas hydrophila. J. General Microbiol. 134: 151-160, 1988.
111.Leung KY, Stevenson MW. Tn5-Induced protease-deficient strains of Aeromonas hydrophila with reduced virulence for fish. Infect Immun. 56: 2639-2644, 1988.
112.Lior H, Johnson WM. Application of the polymerase chain reaction (PCR) to detection of the aerolysin gene in whole cell cultures of β-hemolytic Aeromonas hydrophila. Experientia 47:421-424, 1991.
113.Ljungh A, Kronevi T. Aeromonas hydrophilatoxins: Intestinal fluid accumulation and mucosal injury in animal models. Toxicon. 20: 397-407, 1982.
114.Ljungh A, Wadstrom T. Aeromonas toxins. Pharmacol Ther. 15: 339-354, 1982.
115.Ljungh A, Wadstrom T. Preparation of hemolysin from Aeromonas. Methods Enzymol. 165:200, 1988.
116.Ljungh A, Wretlind B, Mollby R. Separation and characterization of enterotoxin and two hemolysins from Aeromonas hydrophila. Acta Pathol Microbiol Scand Sect. 89: 387-397, 1981.
117.Loewy AG, Santer UV, Wieczorek M, Blodgett JK, Jones SW, Vheronis LC. Purification and characterization of a novel zinc-proteinase from cultures of Aeromonas hydrophila. J Biol Chem. 268: 9071-9078, 1993.
118.Loychern A, Charoensiri K, Ratanabanangkoon K. A simple pruification of Aeromonas hydrophila hemolysin. J Natural Toxins. 3:1-6, 1994.
119.MacIntyre S, Buckley JT. Presence of glycerophospholipid: cholesterol acyltransferase and phospholipase in culture supernatant of Aeromonas hydrophila. J Bacteriol. 135: 402-407, 1978.
120.MacIntyre S, Trust TJ, Buckle JT. Identification and characterization of outer membrane fragments released by Aeromonas spp. Can J Biochem. 58: 1018-1025, 1980.
121.Majeed KN. Mac Rae IC. Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures. Int J Food Microbiol. 12: 181-188, 1991.
122.Mateos D, Anguita J, Naharro G, Paniagua C. Influence of growth temperature on the production of extracellular virulence factors and pathogenicity of environmental and human strains of Aeromonas hydrophila. J Appl Bacteriol. 74: 111-118, 1993.
123.McCardell BA, Madden JM, Kothary MH, Sathyamoorthy V. Purification and characterization of CHO cell-elongating toxin produced by Aeromonas hydrophila. Microb Pathog. 19:1-9, 1995.
124.Merino S, Aguilar A, Nogueras MM, Regue M, Swift S, Thomas JM. Cloning, sequencing and role in virulence of two phospholipases (A 1 and C1) from mesophilic Aeromonas sp. Serogroup O:34. Infect Immun. 67: 4008-4013, 1999.
125.Monfort P, Baleux B. Dynamics of Aeromonas hydrophila, Aeromonas sobria and Aeromonas caviae in sewage treatment pond. Appl Environ Microbiol. 56: 1999-2006, 1990.
126.Myiazaki T, Jo Y. A histopathological study of motile Aeromonad disease in Ayu. Fish Pathology. 20: 55-59, 1985.
127.Namdari H, Bottone EJ. Cytotoxin and enterotoxin production as factors delineating enteropathogenicity of Aeromonas caviae. J. Clin. Microbiol. 28: 1796-1798, 1990.
128.Nelson KL, Raja SM, Buckley JT. Nelson. The glycosylphosphatidylinositol- anchored surface glycoprotein Thy-1 is a receptor for the channel-forming toxin aerolysin. J Biol Chem 272: 12170-12174, 1997.
129.Nieto TP, Corcobado MR, Toranzo AE, Barja JL. Relation of water temperature to infection of Salmo airdneri with motile Aeromonas. Fish Pathology 20: 99-105, 1985.
130.Nieto TP, Ellis K. Characterization of extracelluar metallo-and serine protease of Aeromonas hydrophila strain B51. J Gen Microbiol. 132: 1975-1979, 1986.
131.Nord CE, Sjoberg L, Wadstrom DT, Wretlind B. Characterization of three Aeromonas and nine Pseudomonas species by extracellular enzymes and hemolysins. Med Microbiol Immumol. 161: 79-87, 1975.
132.Parker MW, Buckley JT, Postma JPM, Tucker AD, Leonard K, Pattus F, Tsernoglou D. Structure of Aeromonas toxin proasrolysin in its water-soluble and membrane-channel states. Nature. 367: 292-295, 1994.
133.Parker MW, Tucker AD, Tsernoglou D, Pattus F. Insight into membrane insertion based on studies of colicins. Trends Biochem Sci. 15:126-129, 1990.
134.Pollard DR, Johnson WM, Lior H, Tyler SD, Rozee KR. Detection of the aerolysin gene in Aeromonas hydrophilaby the polymerase chain reaction. J Clin Microbiol. 28: 2477-2481, 1990.
135.Popoff. Aeromonas hpdrophila. In: John GH, ed. Bergey’s Manual of Systematic Bacteriology vol. I. Williams & Wilkins, Baltimore. 545-554, 1984.
136.Rahman AF, Willoughby JM. Dysentery-like syndrome associated with Aeromonas hydrophila. Br Med J 281: 976-977, 1980.
137.Rivero O, Anguita J, Paniagua C, Naharro G. Molecular cloning and characterization of an extracellular protease gene from Aeromonas hydrophila. J Bacteriol. 172: 3905-3908, 1990.
138.Rodriguez LA, Ellis AE, Nieto TP. Purification of an extracellular metalloprotease, serine protease and haemolysin of Aeromonas hydrophila strain B32 all are lethal for fish. Microb Pathogen. 13: 17-24, 1992.
139.Rose JM, Houston CW, Coppenhaver DH, Dixon JD, Kurosky A. Purification and characterization of a cholera toxin-cross-reactive cytolytic enterotoxin produced by a human isolate of Aeromonas hydrophila. Infect Immun. 57: 1165-1169, 1989.
140.Santos Y, Toranzo AE, Barja JL, Nieto TP, Villa TG. Virulence properties and enterotoxin production of Aeromonas strains isolated from fish. Infect Immun. 56: 3285-3293, 1988.
141.Schultz AJ, McCardell BA. DNA homology and immunological cross reactivity between Aeromonas hydrophila cytotonic toxin and cholera toxin. J Clin Microbiol. 26:57-61, 1988.
142.Shaw DH, Squires MJ. O-antigen in a virulent strain of Aeromonas hydrophila. FEMS Microbiol Lett. 24: 277-280, 1984.
143.Shieh HS. Toxicity of Aeromonas hydrophila protease to Atlantic salmon. Microbios Lett. 36: 17-21, 1987.
144.Shotts EB, Gaines JL, Martin L. Aeromonas-induced deaths among fish and reptiles in a eutrophic inland lake. J Am Vet Med Assoc 161: 603-607, 1972.
145.Stephen S, Rao KNA, Kumar MS, Andrani R. Human infection with Aeromonas Species: varied clinical manifestations. Ann Intern Med 83: 368-369, 1976.
146.Thelestam M, Ljungh A. Membrane damaging and cytotoxic effects on human fibroblasts of alpha- and beta- hemolysins from Aeromonas hydrophila. Infect Immun. 34: 949-956, 1981.
147.Thomton J, Howard SP, Buckley JT. Molecular cloning of a phospholipid- cholesterol acyltransferase from Aeromonas hydrophila. Sequence homologies with lecithin-cholesterol acyltransferase and other lipases. Biochim Biophys Acta. 959: 153-159, 1988.
148.Thune RL. Extracellular proteases from Aeromonas hydrophila partial purification and effects on age channel catfish. Trans Am Fish. Soc. 111:749-754, 1982.
149.Thune RL, Graham TE, Riddle LM, Amborski RL. Extracellular protease from Aeromonas hydrophila: partial purification and effects on age-0 channel catfish. Am Fish Soc. 111: 749-754, 1982.
150.Thune RL; Johnson MC; Graham TE, Amborski RL. Aeromonas hydrophilaβ-haemolysin: purification and examination of its role in virulence in O-group channel catfish, Ictalurus punctatus(Rafinesque). J Fish Dis. 9: 55-61, 1986.
151.Thune RL, Stanley LA, Cooper RK. Pathogenesis of gream-negative bacterial infections in warm-water fish. In: Faisal M, ed. Annual Review of Fish Disases. Pergamon Press Co, Oxford, 145-185, 1993.
152.Titball RW. Bacterial Phospholipases C. Microbiological Review, Microbiol Rev. 57: 347-366, 1993.
153.Todd LS, Hardy JC, Stringer MF. Toxin production by strains of Aeromonas hydrophila grown in laboratory media and prawn pure. Int J Food Microbiol. 9: 145-156, 1989.
154.Trust TJ. Pathogenesis of infectious diseases of fish. Ann Rev Microbiol. 40: 479-502, 1986.
155.van der Goot FG, Ausio J, Wong KR, Pattus F, Buckley JT. Dimerization stabilizes the pore-forming toxin aerolysin in solution. J Biol chem. 268: 18272-18279, 1993.
156.van der Goot FG, Pattus F, Parker M, Buckley JT. The cytolytic toxin aerolysin: from the soluble form to the transmembrane channel. Toxicol. 87: 19-28, 1994.
157.van der Goot FG, Pattus F, Wong KR, Buckley JT. Oligomerization of the channel-forming toxin aerolysin precedes insertion into lipid bilayers. Biochemistry. 32: 2636-2642, 1993.
158.Wadstrom T, Ljungh A, Wretlind B. Enterotoxin, hemolysin, and cytotoxic protein in Aeromonas hydrophila from human infections. Acta Pathol Microbiol Scand Sect B. 84:112-114, 1976.
159.Wandersman C, Delepelaire P. To1C, an Escherichia coli outer membrane protein required for hemolysin secretion. Proc Natl Lead Sci USA. 87: 4776-4780, 1990.
160.Wilcox MH, Cook A, Geray I, Eley A. Toxin production, adherence and protein expression by clinical Aeromonas spp. isolates in broth and human pooled ileostomy fluid. Epidemiol Inf. 113: 235-245, 1994.
161.Wretlind B, Heden L, Wadstrom T. Formation of extracellular hemolysin by Aeromonas hydrophila in relation to protease and staphylolytic enzyme. J Gen Microbiol. 78: 57-65, 1973.
162.Wu, H.S., Liu DP, Hwang CH, Chen MJ, Hwang JL, Liu Y, Shankuan LC, Lin CS, Wu TN. Survey on the distribution of Vibrionaceae at the seaport areas in Taiwan, 1991-1994. Chinease J Microbiol Immunol. 29: 197-209, 1996.