數年來石斑魚細菌皮膚及肌肉的出血性潰瘍，推測為病原菌之胞外蛋白酶分解肌肉所造成的。因此本研究調查海洋菌之胞外蛋白酶活性，並分離石斑致病菌，探討其特性。海水背景菌相調查的採樣點為台東外海，使用MB2216 及PPESⅡ兩種培養基。細菌胞外蛋白酶活性分析則採用milk seawater (MS)培養基、azocasein及azoalbumin三種方法，其活性為1~5 U/mg。由MB2216所分離的細菌菌株普遍比PPESⅡ之菌株有較高的平均蛋白酶活性。以azocasein及azoalbumin法來分析蛋白酶活性，所得到的趨勢大致相似，而MS 透明環之結果變異度大。自養殖場石斑魚皮膚所分離FI-02致病株，經過16S rDNA序列分析及生化鑑定後判定為Aeromonas hydrophila，該菌可以生長在含0~4% NaCl的LB中，此菌不被ampillicin、cefazolin、erythomycin、sulfisoxazole所抑制，而對gentamicine、streptomycin、doxycline、norfloxacin、cefotazime、oxolinic acid及tetracycline具感受性。在FI-02的胞外蛋白之蛋白酶活性研究，分別用四種培養基培養FI-02細菌，酵素活性皆在4 U/mg以上，其中以生長在LB培養基上的菌株酵素活性最高，約為6.08 U/mg。以27, 37, 42及56℃加熱二十分鐘後，測定胞外蛋白酶之活性，其結果並無顯著的差異。在魚類抗菌免疫力的研究時，由於血清成分複雜，尚無法找出對抗石斑抗體IgM之二級抗體，仍待將來進一步改進之。

In recent years, the skin and muscle hemorrhagic ulcer occurred in grouper hatchery, suggesting that the bacteria extracellular protease was involved. In this study I surveyed the extracellular protease activity of marine bacteria and investigated the characteristics of the pathogen isolated from diseased grouper. The sampling sites of background survey were located along the coast of Taitung. The cultured media were MB2216 and PPESII. The extracellular protease activity was measured using milk seawater medium (MS) and dye-modified proteins of azocasein and azoalbumin. The extracellular protease activities of the marine bacteria were 1~5 U/mg. The average protease activities of the bacteria from MB2216 media were higher than those from PPESII media. The extracellular protease activity assay using azocasein and azoalbumin had similar tendency but the result by the clear zone measurement on the MS medium was of high variations. The FI-02 isolated from the diseased grouper skin was identified as Aeromonas hydrophila by the biochemicial characteristics and 16S rDNA sequence analysis. FI-02 was able to grow on the LB broth with 0~4% NaCl. The FI-02 was resistant to ampillicin, cefazolin, erythomycin, sulfisoxazole, whereas it was sensitive to streptomycin, doxycline, norfloxacin, cefotazime, oxolinic acid, and tetracycline. When FI-02 growing in the four media, the extracellular protease activities were higher than 4 U/mg. The highest protease activity was 6.08 U/mg when the bacteria grew on the LB medium. After heating on 27, 37, 42 and 56℃ for 20 mins, the protease activities remained the same. In the investigation of the fish immunity, owing to the complex compositions of the fish serum, no conclusion can be drawn on the specificity of the antibody against the bacterium.

謝誌 ----------------------------------------Ⅰ
中文摘要 ------------------------------------Ⅱ
Abstract ------------------------------------Ⅲ
目錄 ----------------------------------------Ⅳ
表目錄 --------------------------------------Ⅴ
圖目錄 --------------------------------------Ⅵ
壹、前言
一、 台灣養殖石斑魚概況 ----------1
二、 魚病的診斷 ------------------3
三、 海洋常見致病菌 --------------6
四、 研究目的 --------------------21
貳、材料與方法
一、 海洋菌之培養 -----------------22
二、 石斑魚致病菌之分離 -----------22
三、 細菌胞外產物之製備
(extracellular products, ECP)----23
四、 蛋白酶 (protease) 活性測定 ----23
五、 蛋白質濃度定量 ----------------24
六、 革蘭氏染色 --------------------25
七、 菌種鑑定 ----------------------25
八、 細菌生理特性分析 -------------28
九、 細菌對石斑魚之攻擊試驗 -------30
十、 石斑魚免疫球蛋白之偵測 -------31
參、結果 ----------------------------------------38
肆、討論 ---------------------------------------45
伍、參考文獻 ---------------------------------52
陸、圖表 -------------------------------------76
附錄A ---------------------------------------107
附錄B ---------------------------------------108
附錄C ---------------------------------------112

王孟群。1983。實用微生物學實驗。
安可倫。1969。鰻魚鰭紅病之防治試驗。中國水產。pp.14-22。
林育興、張永昌、劉正義、王渭賢。1997。甲魚親水性產氣單菌感染症。魚病研究專輯。18: 51-57。
林曜松，蕭世民。1977。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。魚病研究專輯(一): 57-61。
呂明偉。2002。龍膽石斑神經壞死病毒外殼蛋白與其似病毒顆粒之研究。國立中山大學海洋資源研究所，博士論文。
呂學群。2003。產氣單胞菌對養殖香魚之致病性及病理學研究。國立海洋大學水產養殖學研究所，碩士論文。
郭光雄，鍾虎雲。養殖池之R+抗藥細菌。1987。農委會漁業特刊第十號，魚病研究論文集(一)細菌及免疫專輯。pp.70-77。
莊文效。2001。弧菌Vibrio carchariae引起養殖石斑魚腸胃症(gastroenteritis)之致病機制研究。國立海洋大學水產養殖學研究所，碩士論文。
陳昭德，郭光雄。1978。養殖鰻魚細菌分布之研究。魚病研究專輯(二): 15-32。
陳林陽，簡秋源。臺灣養殖黑鯛爛尾病病原菌Aeromonas hydrophila之分離。1987。農委會漁業特刊第十號，魚病研究論文集(一)細菌及免疫專輯。pp. 78-91。
陳瑤湖。1999。水產養殖概況。行政院勞工委員會職業訓練局。
曾文陽。1987。石斑魚養殖學。前程出版社。
黃卓男。2001。香魚分出親水性產氣單胞菌(Aeromonas hydrophila)重組aerolysin之表現與特性。國立台灣大學獸醫學研究所。碩士論文 。
黃世鈴、陳秀男。2000。魚病的診斷及防治(十)。漁業推廣，164期。
黃貴民。1999。實用石斑魚養殖。水產出版社。
葉玉萍。1999。溶藻弧菌及哈威弧菌對黃鰭鯛之病源性研究。。國立海洋大學水產養殖學研究所，碩士論文。
鄭敏孚。1997。腸炎弧菌疑似致病因子之純化與特性分析。國立台灣大學農業化學研究所，碩士論文。
劉富光。臺灣養殖泥鰍之疾病。1979。中國水產324: 14-18。
蕭世民，陳孝禹。1977。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。pp. 19-21。
鍾虎雲。1983。談談魚病。科學月刊，0160期。
鍾虎雲，郭光雄。1973。魚體常有細菌之研究-Ⅰ存在外觀健康鰻魚之鰓、腸血液及內臟之細菌。臺灣水產學會刊。2(2): 20-25。
簡肇衡、賈愛華。1994。赤鰭病原菌鰻之病理生理之研究。魚病研究專輯。14: 13-24。
魏紋祈，2000。封閉式草蝦養殖池細菌族群之研究。國立中山大學海洋資源所，碩士論文。
魏榮璋，1998。嗜水性產氣單胞菌對泰國長臂大蝦之致病性研究。國立海洋大學水產養殖研究所，碩士論文。
Abeyta, C. A., M. M. Wekell, J. J. Sullivn, and G. N. Stelma. 1986. Recovery of Aeromonas hydrophila from oysters implicated in an outbreak of foodborne illness. J. Food Prot. 49: 643-646.
Acuigrup. E. 1980. Flavobacteriosis in coho salmon (Oncorhynchus kisutch). In: Ahne, W. (ed), Fish Diseases, Third COPRAQ-Session, Berlin, Springer-Verlag. pp. 212-217.
Adams, D., H. M. Atkinson, and W. H. Woods. 1983. The differing effect of protease on the adhesions of Aeromonas hydrophila. FEMS Microbiol. Lett. 20: 197-200.
Agger, W. A., J. D. McCormick, and M. J. Gurwith. 1985. Clinical and microbiological features of Aeromonas hydrophila-associated diarrhea. J. Clin. Microbiol. 21: 909-913.
Ahne, W., W. Popp, and R. Hoffmann. 1982. Pseudomonas fluorescens as a pathogen of tench (Tinca tinca). Bulletin of the European Association of Fish Pathologists. 4: 56-57.
Akazawa, H. 1968. Bacterial disease of marine fishes. Bulletin of the Japanese Society of Scientific Fisheries. 34: 271-272.
Allan, B. and M. W. Stevenson. 1981. Extracellular virulence factors of Aeromonas hydrophila in fish infections. Can. J. Microbiol. 27: 1114-1122.
Altwegg, M., A. G. Steigerwalt, R. Altwegg-Bissig, J. Luthy-Hottenstein, and D. J. Bremmer. 1990. Biochemical identification of Aeromonas
genospecies isolated from humans. J. Clin. Microbiol. 28: 258-264.
Amin, N. E., I. S. Abdallah, T. Elallawy, and S. M. Ahmed. 1985. Motile Aeromonas septicaemia among Tilapia nilotica (Sarotherodon niloticus) in upper Egypt. 20: 93-97.
Aoki, T. 1988. Drug-resistant plasmids from fish pathogens. Microbiol. Sci. 5: 219-223.
Austin, B. and D. A. Austin. 1987. Aeromonads. In: Austin, B., Austin, D. (Eds.), Bacterial Fish Pathogens: Diseases in Farmed and Wild Fish. Ellis Horwood. Toronto.
Austin, B. and D. A. Austin. 1993. Bacterial Fish Pathogens: Diseases in Farmed and Wild Fish. 2nd end. Ellis Horwood. Limited, Chichester, pp. 359-373.
Azad, I. S., K. M. Shankar, C. V. Mohan, and B. Kalita. 1999. Biofilm vaccine of Aeromonas hydrophila-standardization of dose and duration for oral vaccination of carps. Fish and Shellfish Immunology. 9: 519-528.
Barham, W. T., H. Schoonbee, and G. L. Smit. 1979. The occurrence of Aeromonas and Streptococcus in rainbow trout (Salmo gairdneri). Journal of Fish Biology. 15: 457-460.
Barton, T. A., L. A. Bannister, S. G. Griffiths, and W. H. Lynch. 1997. Further characterization of Renibacterium salmoninarum extracellular products. Appl. Environ. Microbiol. 63: 3770-3775.
Baumann, P. A., L. Furniss, and J. V. Lee. 1984. Vibrio. In: Krieg, N. R. (Ed.), Bergey’s Mannual of Systematic Bacteriology. Vol. 1. Williams and Wilkins, Baltimore, pp.518-544.
Becker, C. D. and M. P. Fujiwaha. 1978. The Bacterial Pathologen Flexibacter columnaris and its Epizootiology among Columbia River Fish. Monograph No.2, Washington, DC, American Fisheries Society.
Bein, S. J. 1954. A study of certain chromogenic bacteria isolated from ‘red-tide’ water with a description of a new species. Bulletin of Marine Science in the Gulf and Caribbean. 4: 110-119.
Bejerano, Y., S. Sarig, M. T. Horne, and R. J. Roberts. 1979. Mass mortalities in silver carp Hypophthalmichthys molitrix (Valenciennes) associated with bacterial infection following handling. J. Fish Dis. 2: 49-56.
Beynon, R. J. and J. S. Bond. 1990. Proteolytic enzymes-a practical approach. IRL Press.
Bloch, S. and H. Monteil. 1989. Purification and characterization of Aeromonas hydrophila beta-hemolysin. Toxicon. 27: 1279-1287.
Bulkock, G. L. 1981. Strptococcal infections of fishes. United States Department of the Interior, Fish and Wildlife Service, Fish Disease Leaflet. 63: 1-7.
Bullock, G. L. and S. F. Snieszko. 1970. Fin rot, coldwater disease, and pseduncle disease of salmonid fish. United States Department of the Interior, Fish and Wildlife Service, Fish Disease Leaflet. 25: 1-3.
Burke, J. and L. Rodgers. 1981. Idenification of pathogenic bacteria associated with the occurrence of ‘red-spot’ in sea mullet, Mugil cephalus L., in south-eastern Queensland. J. Fish Dis. 3: 153-159.
Burke, V., J. Robinson, R. J. Beny, and M. Gracey. 1981. Detection of enterotoxins of Aeromonas hydrophila by a suckling-mouse test. J. Med. Microbiol. 14: 401-408.
Burke, V., J. Robinson, M. Gracey, D. Peterson, and K. Partidge. 1984. Isolation of Aeromonas hydrophila from a metropolitan water supply: Seasonal correlation with clinical isolates. Appl. Environ. Microbiol. 48: 361-366.
Burno, D. W. 1988. The relationship between autoagglutination, cell sueface hydrophobicity and virulence of fish pathogen Renibacterium salmoninarum. FEMS Micrboiol. Lett. 58: 135-140.
Burno, D. W. 1990. Presence of a saline-extractable protein associated with virulence strains of the fish pathogen Renibacterium salmoninarum. Bull. Eur. Assoc. Fish Pathol. 10: 8-10.
Carson, J., T. Wagner, T. Wilson, and L. Donachie. 2001. Miniaturized tests for computer-assisted identification of motile Aeromonas species with an improved probavility matrix. J. Appl. Microbiol. 90: 190-200.
Chabot, D. J. and R. L. Thune. 1991. Protease of the Aeromonas hydrophila complex: identification, characterization and relation to virulence in channel catfish, Ictalurus punctatus (Rafinesque). J. Fish Dis. 14: 171-183.
Chakraborty, T., M. A. Montenegro, S. C. Sanyal, R. Helmuth, E. Bulling, and K. N. Timmis. 1984. Cloning of enterotoxin gene from Aeromonas hydrophila provides conclusive of production a cytotonic enterotoxin. Infect. Immun. 46: 435-441.
Chakraborty, T., B. Huhle, H. Bergbaure, and W. Goebel. 1986. Cloning, expression and mapping of the Aeromonas hydrophila aerolysin gene determinant in E. coli K-12. J. Bacteriol. 167: 368-374.
Chen, F. R., P. C. Liu, and K. K. Lee. 2000. Lethal attribute of serine protease secreted by Vibrio alginolyticus strains in kuruma prawn Penaeus japonicus. Z. Naturforsch. 55: 94-99.
Chen, S. N., S. L. Huang, and G. H. Kou. 1992. Studies on the epizootiology and pathogenicity of bacterial infections in cultured giant tiger prawn, Penaeus monodon in Taiwan. In Diseases of Cultured Penaeid Shrimp in Asia and the United States ed. Fulks, W. and Main, K. L. pp.195-205. Hawaii: The Oceanic Institute.
Cheronis, J. C. and J. E. Repine. 1993. Protease, Protease Inhibitors, Protease Derived peptides: Importance in human pathophysiology and therapeutics. Birkh auser, Boston. pp.3-18.
Chopra, A. K., C. W. Houston, C. T. Genaux, J. D. Dixon, and A. Kurosky, 1986. Evidence for production of an enterotoxin and cholera toxin cross reactive factor by Aeromonas hydrophila. J. Clin. Microbiol. 24: 661-664.
Cipriano, R. C. 2001. Revision of Fish Diseases Leaflet 68 (1984), “Aeromonas hydrophila and Motile Aeromonad Septicemias of Fish.” By R. C. Cipriano, G. L. Bullock, and S. W. Pyle.
Coleman, G. and P. W. Whitby. 1993. A comparison of the amino acid sequence of the serine protease of the fish pathogen Aeromonas salmonicida subsp. salmonicida with those of other subtilisin-type enzymes relative to their substrate-binding sites. J. Gen. Microbiol. 139: 245-249.
Colorni, A., I. Paperna, and H. Gordin. 1981. Bacterial infections in gilthead sea bream Sparus aurata cultured at Elat. Aquaculture. 23: 257-267.
Colwell, R. R. and D. J. Grimes. 1984. Vibrio diseases of marine fish populations. Helgolander Meeresuntersuchungen. 37: 265-287.
Colwell, R. R., M. R. MacDonell, and J. Deley. 1986. Proposal to recognize the family Aeromonadaceae fam. Int. J. Syst. Bacteriol. 36: 473-477.
Cone, D. K. 1982. A Lactobacillus sp. from diseased female rainbow trout, Salmo gairdneri Richardson, in Newfoundland, Canada. J. Fish. Dis. 5: 479-485.
Cruz, J. M., A. Saraiva, J. C. Eiras, R. Branco, and J. C. Sousa. 1986. An outbreak of Plesiomonas shigelloides in farmed rainbow trout, Salmo gairdneri Richardson, in Portugal. Bulletin of the European Association of Fish Pathologists. 6: 20-22.
Daly, J. G. and R. M. W. Stevenson. 1987. Hydrophobic and hemoglutinating of a cytotonic produced by Aeromonas hydrophila. Can. J. Microbiol. 133: 3575-3580.
Dawson, M. P., B. A. Humphrey, and K. C. Marshall. 1981. Adhesion: A tactic in the survival strategy of a marine Vibrio during starvation. Curr. Micrbiol. 6: 195-199.
Doetch, R. H. 1981. Determative methods of light microbiology. In: R. G. E. Murray, R. N. Costilow, E. W. Nester, W. A. Wood, N. R. Krieg and G. B. Phillips (Editors), Manual of methods for general bacteriology. Am. Soc. Microbiol., Washington D C. pp. 21-33.
Doing, P., L. Emody, and T. J. Trust. 1992. Binding of laminin and fibronectin by the trypsin-resistant major structural domain of the crystalline virulence surface array protein of Aeromonas salmonicda. J. Biol. Chem. 267: 43-49.
Donta S. T. and A. D. Haddow. 1978. Cytotoxic activity of Aeromonas hydrophila. Infect. Immun. 21: 989-993.
Dooley, J. S. G., R. Lallier, D. H. Shaw, and T. J. Trust. 1985. Electrophoretic and immunochemical analyses of the lipopolysaccharides from Aeromonas hydrophila. J. Bacteriol. 164: 263-269.
Dulin, M. P. 1979. A review of tuberculosis (mycobacteriosis) in fish. Veterinary Medicine/Small Animal Clinican. pp. 735-737.
Ellis, A. E. 1991. An appraisal of the extracellular toxin of Aeromonas salmonisida sp. J. Fish. Dis. 14: 265-277.
Ellis, A. E., A. do Vale, T. J. Bowden, K. Thompson, and T. S. Hastings. 1997. In vivo production of A-protein, lipopolysaccharide, iron-regulated outer membrane proteins and 70 kDa serine protease by Aeromonas salmonicida subsp. salmonicida. FEMS. Microbiol. Lett. 149: 157-163.
Farrell, D. H. and J. H. Crosa. 1991. Purification and characterization of a secreted protease from the pathogenic marine bacterium Vibrio anguillarum. Biochemistry. 30: 3432-3436.
Farto, R., M. J. Perez, A. Fernandez-Briera, and T. P. Nieto. 2002. Purification and characterization of a fish lethal extracellular protease from Vibrio pelagius. Vet. Microbiol. 89: 181-194.
Fernandez, L., P. Secades, J. R. Lopez, I. Marquez, and J. A. Guijarro. 2002. Isolation and analysis of a protease gene with an ABC transport system in the fish pathogen Yersinia ruckeri: insertional mutagenesis and involvement in virulence. Microbiol. 148: 2233-2243.
Freij, B. J. 1984. Aeromonas: Biology of the organism and disease in children. Pediatr. Infect. Dis. J3: 164-175.
Fryer, J. L. and J. E. Sanders. 1981. Bacterial kidney diseases of salmonid fish. Ann. Rev. Microbio. 35: 273-298.
Fuhrmann, H., K. H. Bohm, and H. J. Schlotfeldt. 1983. An outbreak of enteric redmouth diseases in West Germany. J. Fish. Dis. 6: 309-311.
Fyfe, L., G. Coleman, and A. L. Munro. 1987. Identification of major common extracellular proteins secreted by Aeromonas salmonicida strains isolated from diseased fish. Appl. Environ. Microbiol. 53: 722-726.
Goguen, J. D., N. P. Hoe, and Y. V. B. K. Subrahmanyam. 1995. Protease and bacterial virulence: a view from the trenches. Infect. Agents Dis. 4: 47-54.
Gray, S. J. 1984. Aeromonas hydrophila in livestock incidence biochemical characteristics and antibiotic susceptibility. J. Hyg. 92: 365-376.
Grayson, T. H., A. J. Evenden, M. L. Gilpin, K. L. Martin, and C. B. Munn. 1995. A gene from Renibacterium salmoninarum encoding a product which shows homology to bacterial zinc-metalloproteases. Microbiology. 141: 1331-1341.
Grimes, D. J., R. R. Colwell, J. Stemmler, H. Hada, D. Maneval, F. M. Hetrick, E. B. May, R. T. Jones, and M. Stoskopf. 1984. Vibrio species as agents of elasmobranch diseases. Helgolander Meeresuntersuchungen. 37: 309-315.
Grisez, L., R. Ceusters, and F. Ollevier. 1991. The use of API 20E for the identification of Vibrio anguillarum and V. ordalii. J. Fish Dis. 14: 359-365.
Hawke, J. P. 1979. A bacterium associated with disease of pond cultured channel catfish, Ictalurus punctatus. J. Fish. Res. Board. Canada. 36: 1508-1512.
Herman, R. L. 1968. Fish furunculosis 1952-1966. Transactions of the American Fisheries Society. 97: 221-230.
Hester, F. E. 1973. Fish health: a nationwide survey of problems and needs. Progressive Fish Culturist. 35: 11-18.
Hijltnes, B., K. Anderson, and E. Egidius. 1987. Multiple antibiotic resistance to Aeromonas salmonicida. Bull. Eur. Ass. Fish Pathol. 7: 85.
Hiu, S. F., R. A. Holt, N. Sriranganathan, R. J. Seidler, and J. L. Fryer. 1984. Lactobacillus piscicola, a new species from salmonid fish. Int. J. Sys. Bacterio. 34: 393-400.
Hokama, A., Y. Honma, and N. Nakasone. 1990. Pili of an Aeromonas strain as a possible colonization factor. Microbiol. Immun. 34: 901-915.
Holt, J. G., N. R. Krieg, P. H. A. Sneath, J. T. Staley, and S. T. Williams. 1994. In Bergey’s Mannual of Determinative Bacteriology, 9th ed. William and Wilkins, Baltimore, pp. 192-289.
Howard, S. P., W. J. Garland, M. J. Green, and J. T. Buckley. 1987. Nucleotide sequence of the gene for the hole-forming toxin aerolysin of Aeromonas hydrophila. J. Bacteriol. 169: 2869-2871.
Huizinga, H. W., G. W. Esch, and T. C. Hazen, 1979. Histopathology of red-sore disease (Aeromonas hydrophila) in naturally and experimentally infected largemouth bass Micropterus salmoides (Lacepede). J. Fish. Dise. 2: 263-277.
Igbal, M. M., K. Tajima, K. Nakano, and T. Ezura. 1998. Phenotypic and genotypic identification of motile Aeromonads isolated from fish with epizootic ulcerative syndrome in Southeast Asian Countries. Fish Pathol. 33: 255-263.
Inglis, V., R. J. Roberts, and N. R. Bromage. 1993. Bacterial Diseases of Fish., Blackwell Scientific Publications, Oxford. pp. 312.
Janda, J. M. 1991. Recent advances in the study of taxonomy, pathogenicity and infectious syndromes associated with the genus Aeromonas. Clin. Microbiol. Rev. 4: 397-410.
Janda, J. M. and S. L. Abbott. 1998. Evolving concepts regarding the genus Aeromonas: an expanding panorama of species, disease presentation and unanswered questions. Clin. Infect. Dis. 27: 332-344.
Janda, J. M., L. S. Guthertz, R. P. Kokka, and T. Schimada. 1994. Aeromonas species in septicemia: laboratory characteristics and clinical observations. Clin Infect Dis. 19: 77-83.
Johnson, W. M. and H. Lior. 1981. Cytotoxicity and suckling mouse reactivity of Aeromonas hydrophila isolated from human sources. Can. J. Microbiol. 27: 1019-1027.
Jukes, T. H. and C. R. Cantor. 1969. Evolution of protein molecules. In: H. N. Munro.(ed.). Mammalian protein metabolism. Vol. 3. Academic Press., Inc. New York.
Kampfer, P. and M. Alwegg. 1992. Numerial classification and identification of the motile Aeromonas species. Ann. Rev. Fish Dis. 4: 315-343.
Kanai, K. and Y. Takagi. 1986. Alpha-hemolytic toxin of Aeromonas hydrophila produced in vivo. Fish Pathology. 21: 245-250.
Kay, B. A., C. E. Guerrero, and R. B. Sack. 1985. Media for the isolation of Aeromonas hydrophila. J. Clin. Microbiol. 22: 888-890.
Keusch, G. T. and S. T. Donta. 1975. Classification of enterotoxins in the basis of activity in cell culture. J. Infect. Dis. 131: 58-63.
Khardori, N. and V. Faonstein. 1988. Aeromonas and Plesiomonas as etiological agent. Ann. Rev. Microbiol. 42: 395-419.
Krovacek, K., K. Huang, S. Sternberg, and S. B. Svenson. 1998. Aeromonas hydrophila septicaemia in a grey seal (Halichoerus grypus) from the Baltic Sea: a case study. Com. Immunol. Microbiol. & Infect. Dis. 21: 43-49.
Kubota, S. S., N. Kaige, and T. T. Miyashita. 1981. Histopathological stydies on edwardisellosis of Tilapia-1. Natural infection. Bulletin of Faculty of Fisheries, Mie University. 9: 155-165.
Kuijper, E. J., A. G. Steigerwalt, B. S. C. I. M. Schoenmakers, M. F. Peeters, H. C. Zanen, and D. J. Brenner. 1989. Phenotypic characterization and DNA relatedness in human fecal isolates of Aeromonas spp. J. Clin. Microbiol. 27: 132-138.
Kuijper, E. J., L. V. Alphen, M. F. Peeters, and D. J. Brenner. 1990. Human serum antibody reponse to the presence of Aeromonas spp. in the intestinal tract. J. Clin. Microbiol. 28: 584-590.
Kusuda, R. and A. Sugiyama. 1981. Studies on the characters of Staphylococcus epidermidis isolated from diseased fishes-1. On the morphological, biological and biochemical properties. Fish Pathology. 16: 15-24.
Kusuda, R. and M. Yamaoka. 1972. Etiological studies on bacterial pseudotuberculosis in cultured yellowtail with Pasteurella piscicda as the causative agent-Ⅱ. On the serological properties. Fish Pathology. 13: 79-83.
Lamas, J., R. Anadon, S. Devesa, and A. E. Toranzo. 1990. Visceral neoplasia and epidermal papillomas in cultured turbot Scophthalmus maximus. Dis. Aquat. Org. 8: 179-187.
Lee, K. K. 1995. Pathogenesis studies in Vibrio alginolyticus in the grouper, Epinephelus malabaricus, Bloch et Schneider. Microbiol. Pathogen. 19: 39-48.
Lee, K. K. and A. E. Ellis. 1989. The quantitative relationship of lethality between extracellular protease and hemolysin of Aeromonas salmonicida in Atlantic salmon, Salmo salar. FEMS Microbiol. 61: 127-132.
Lee, K. K. and A. E. Ellis. 1990. Glycerophospholipid: cholesterol acyltransferase complexed with lipopolysaccharide (LPS) is a major lethal exotoxin and cytolysin of Aeromonas salmonicida: LPS stabilizes and enhances toxicity of the enzyme. J. Bacteriol. 172: 5385-5393.
Lee, K. K. and A. E. Ellis, 1991. The role of the lethal extracellular cytolysin of Aeromonas salmonicida in the pathology of furunculosis. J. Fish Dis. 172: 5385-5393.
Leung, K. Y. and M. W. Stevenson. 1988a. Characteristics and distribution of extracellular proteases from Aeromonas hydrophila. J. Gen. Microbiol. 134: 151-160.
Leung, K. Y., and M. W. Stevenson. 1988b. Tn5-Induced protease-deficient strains of Aeromonas hydrophila with reduced virulence for fish. Infect. Immun. 56: 2639-2644.
Leung, K. Y., L. S. Wong, K. W. Low, and Y. M. Sin. 1997. Mini-Tn5 induced growth- and protease-deficient mutants of Aeromonas hydrophila as live vaccines for blue gourami, Trichogaster trichopterus (Pallas). Aquaculture. 158: 11-22.
Lin, C.-S., M.-W. Lu, L. Tang, W. Liu, C.-B. Chao, C. J. Lin, N. K. Kirshna, J. E. Johnson, and A. Schneemann. 2001. Characterization of virus-like-particles assembled in recombinant baculovirus system expressing the capsid protein of a fish nodavirus. Virology. 290: 50-58.
Lio-Po, G. D., L. J. Albright, and E. V. Alapide-Tendencia. 1990. Aeromonas hydrophila in the epizootic ulcerative syndrome (EUS) of snakhead, Ophicephila striatus and catfish, Clarias batrachus: quantitative estimation in natural infection and experimental induction of sermo-mascular necrotic lesion, In“ Diseases in Asian aquaculture I”(ed. By M. Shariff, R. P. Subasinghe and J. R. Arthur). Fish Health Section, Asian Fisheries Society, Manila: 461-474.
Ljungh, A. and T. Kronevi. 1982. Aeromonas hydrophila toxins: Intestinal fluid accumulation and mucosal injury in animal models. Toxicon. 20: 397-407.
Ljungh, A. and T. Wadstrom. 1981. Aeromonas toxins. Pharmac. Ther. 15: 339-354.
Loghothetis, P. N. and B. Austin. 1994. Immune response of rainbow trout (Oncorhynchus mykiss, Walbaum) to Aeromonas hydrophila. Fish and Shellfish Immunology. 4: 239-254.
Love, M., D. Teebken-Fisher, J. E. Hose, J. J. Farmer, F. W. Hickman, and G. R. Fanning. 1981. Vibrio damsela, a marine bacterium, causes skin ulcers in the damselfish Chromis punctipinnnis. Science. 214: 1139-1140.
Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Rall. 1951. Protein measurement with the folin phenol reagent. J. Boil. Chem. 193: 265-275.
Macintosh, D. J. and M. J. Phillips. 1986. The contribution of environment factors to the ulcerative diseases condition in South East Asia. In” Field and laboratory investigations into ulcerative fish diseases in the Asia-Pacific region” (ed. By R. J. Roberts). FAO. Bangkok. pp.175-207.
McCarthy, D. H. 1975. Fish furunculosis. Journal of Institute of Fisheries Managament .6: p13-18.
Mellergaard, S. 1983. Purification and characterization of a new proteolytic enzyme production by Aeromonas salmonicida. J. Appl. Bacteriol. 54: 289-294.
Meyer, F. P. and G. L. Bullock. 1973. Edwardsiella tarda, a new pathogen of channel catfish (Ictalurus punctatus). Appl. Microbiol. 25: 155-156.
Millership, S. E. and B. Chattopadhyay. 1984. Methods for isolation of Aeromonas hydrophila and Plesiomonas shigelloides from faces. J. Hyg. Vamb. 92: 145-152.
Milton, D. L., A. Norqvist, and H. Wolf-Watz. 1992. Cloning of metalloprotease gene involved in the virulence mechanism of Vibrio anguillarum. J. Bacteriol. 174: 7235-7244.
Mitchell, A. J., J. E. Plumb. 1980. Toxicity and efficacy of Furanace on channel catfish infected experimentally with Aeromonas hydrophila. J. Fish Dis. 3: 93-100.
Morita, Y., Q. Hasan, T. Sakaguchi, Y. Murakami and K. Yokoyama. 1998. Properties of a cold-active protease from psychrotrophic Flavobacterium balustinum P104. Appl. Microbiol. Biotechnol. 50: 669-675.
Morison, D., J. Cornick, G. Shum, and B. Zwicker. 1981. Microbiology and histopathology of ‘saddleback’ disease of underyearling Atlantic salmon, Salmo salar. J. Fish Dis. 4; 243-258.
Moyer, N. P. 1987. Clinical significance of Aeromonas species isolated from patients with diarrhea. J. Clin. Microbiol. 25: 2044-2048.
Mulla, R. and S. Millership. 1993. Typing of Aeromonas spp. By numerical analysis of immunoblotted SDS-PAGE gels. J. Med. Microbiol. 39: 325-333.
Munn, C. B., E. E. Ishiguro, W. W. Kay, and T. J. Trust. 1982. Role of surface components in serum resistance of virulence Aeromonas salmonicida. Infect. Immun. 36: 1069-1075.
Munro, A. L. S., T. S. Hasting, A. E. Ellis, and J. Liversidge. 1980. Studies on an ichthyotoxic material produced extracellularly by the furnculosis bacterium, Aeromonas salmonicida. In: Proceedings of the third session of the EIPAC (FAO/OIE) CPRA. Fish Disease. pp.98-106. Berlin: Springer-Verlag KG.
Muriga, K., S. Takahashi, and H. Yamanoi. 1979. Non-cholera Vibrio isolated from diseased ayu. Bulletin of the Japanese Society of Scientific Fisheries. 45: 829-834.
Myizaki, T., Y. Jo. 1985. A histopathological study of motile Aeromonad diseases in Ayu. Fish Pathology. 20: 55-59.
Myizaki, T., Y. Jo, S. S. Kubota, and S. Egusa. 1977. Histopathological studies in vibriosis of the Japanese eel Anguilla japonica. Part1. Natural infection. Fish pathology. 12: 163-170.
Newman, S. G. 1993. Bacterial vaccines for fish. Annual. Review. of Fish Diseases Pergamon, Oxfor. pp.145-185.
Newman, S. G. and J. J. Majnarich. 1982. Direct immersion vaccination of juvenile rainbow trout, Salmo gairdneri Richardson and juvenile coho salmon, Oncorhynchus kisutch (Walbaum), with a Yersinia ruckeri bacterin. J. Fish. Dis. 5: 229-341.
Nieto, T. P., M. R. Corcobado, A. E. Toranzo, and J. L. Barja. Relation of water temperature to infection of Salmo airdneri with motile Aeromonas. Fish Pathology. 20: 99-105.
Nieto, T. P. and K. Ellis. 1986. Characterization of extracellular metallo-and serine protease of Aeromonas hydrophila strain B51. J. Gen. Microbiol. 132: 1975-1979.
Noga, E. J. 1996. Motile aeromonad infection. In: Noga, E. J. (Ed.), Fish diseases: Diagnosis and Treatment. Mosby-Year Book, St. Louis, pp.141-142.
Nord, C. E., L. Sjoberg, D. T. Wadstrom, and B. Wretlind. 1975. Characterization of three Aeromonas and nine Pseudomonas species by extracellular enzymes and hemolysins. Med. Microbiol. Immunol. 161: 79-87.
Norqvist, A., B. Norrman, and H. Wolf-Watz. 1990. Identification and characterization of a zinc-metalloprotease associated with invasion by the fish pathogen Vibrio anguillarum. Infect. Immun. 58: 3731-3736.
Oakey, H., J. Ellis, and L. Gibson. 1996. A biochemical protocol for the differentiation of current genomospecies of Aeromonas. Zentrablatt fiir Bokteriologie. 284: 32-46.
Pacha, R. E. and E. J. Ordal. 1970. Myxobacterial diseases of salmonids. In: Snieszko, S. F. (ed.), A. Symposium on Diseases of Fishes and Shellfishes, Washington DC, American Fisheries Society, Special Publication. 5: 243-257.
Palleroni, N. J. 1984. Genus 1 Pseudomonas Migual 1894 237AL (Nom. Cons. Opin. 5, Jud. Comm. 1952, 237). In: Krieg, N. R. (ed.) Bergey’s Manual of Systematic Bacteriology, Vol. 1, Baltimore, Williams and Wilkins. pp.141-199.
Plumb, J. A., J. H. Schachte, J. L. Gaines, W. Peltier, and B. Carrol. 1974. Streptococcus sp. From marine fishes along the Alabama and northwest Florida coast of the Gulf of Mexico. Transactions of the American Fisheries Society. 103: 358-361.
Popoff, M. 1984. Aeromonas. In: Krieg, N.R. (Ed.), Bergey’s Manual of Systematic Bacteriology. Vol 1. Williams and Wilkins, Baltimore, pp. 545-548.
Post, G. 1966. Reponse of rainbow trout (Salmo gairdneri) to antigens of Aeromonas hydrophila. J. Fish. Rese. Board. Canada. 23: 1487-1494.
Rasheed, V. M. and J. A. Plumb, 1984. Pathogenicity of non-haemolytiv group B Streptococcus sp. in gulf killfish (Fundulus grandis Baird and Girard). Aquacture, 37: 97-105.
Ransom, D. P. 1978. Bacteriologic, immunologic and pathologic studies of Vibrio sp. Pathogenoic to salmonids. Ph. D. thesis, Oregon State University, Corvallis.
Ransom, D. P., C. N. Lannan, J. S. Rohovec, and J. L. Fryer. 1984. Comparison of histopathology caused by Vibrio anguillarum and Vibrio ordalii and tree species of Pacific salmon. J. Fish. Dis. 7: 107-115.
Rivero, O., J. Anguita, C. Paniagua, and G. Naharro. 1990. Molecular cloning and characterization of an extracellular protease gene from Aeromonas hydrophila. J. Bacteriol. 172: 3904-3908.
Roald, S. O. and T. Hastein. 1980. Infection with an acinetobacter-like bacterium in Atlantic salmon (Salmo salar) broodfish. In Ahne, W. (ed), Fish Diseases, Thrid COPRAQ-Session. Berlin, Springer-Verlag. pp. 154-156.
Robinson, J. A. and F. P. Meyer. 1966. Streptococcal fish pathogen. Journal of Bacteriology. 92: 512.
Rodriguez, L. A., A. E. Ellis, T. P. Nieto. 1992. Purification of an extracellular metalloprotease serine protease and haemolysin of Aeromonas hydrophila strain B32 all are lethal for fish. Micro. Pathogen. 13: 17-24.
Rogers, W. A. and D. Xu. 1992. Protective immunity induced by a commercial Vibrio vaccine hybrid striped. J. Aquat. Anim. Hwal. 4: 303-305.
Roberts, R. J. 1989. Fish pathology, 2nd ed. Bailliere Tindall, London, pp.466.
Ruangpan, L. and Kitao, T. 1991. Vibrio bacteria isolated from black tiger shrimp, Penaeus monodon Fabricius. J. Fish Dis. 14: 383-388.
Saitous, N. and M. Nei. 1987. The neighbor-joining method: a new method for construction phylogentic trees. Mol. Biol. Evol. 4: 406-425.
Sambrook, J. E., F. Fritsch, and T. Maniatis. 1989. Molecular cloning: a laboratory manual. 2nd ed. Cold spring Harbor Laboratory Press. N. Y.
Santo, Y., I. Bandin, S. Nunez, M. Montero, A. Silva, and A. E. Toranzo, 1992. Comparsion of the extracellular biological activities of Vibrio anguillarum and Aeromonas hydrophila. Aquaculture. 107: 259-270.
Santo, Y., A. E. Toranzo, J. L. Barja, T. P. Nieto, and T. G. Villa. 1988. Virulence properties and enterotoxin production of Aeromonas strains isolated from fish. Infect. Immun. 56: 3285-3293.
Schaperclaus, W. 1979. Fishkrankheiten. Berlin, Akademie Verlag.
Schultz, A. J., M. J. McCardell. 1988. DNA homology and immunological cross reactivity between Aeromonas hydrophila cytotonic toxin and cholera toxin. J. Clin. Microbiol. 26: 57-61.
Secades, P., B. Alvarez, and J. A. Guijarro. 2001. Purification and characterization of a psychrophilic, calcium-induced, growth-phase-dependent metalloprotease from the fish pathogen Flavobacterium psychrophilum. Appl. Environ. Microbiol. 67: 2436-24.
Secades, P., B. Alvarez, and J. A. Guijarro. 2003. Purification and properties of a new psychrophilic metalloprotease (Fpp2) in the fish pathogen Flavobacterium psychrophilum. FEMS Microbiol. Lett. 226: 273-279.
Secades, P. and J. A. Guijarro. 1999. Purification and characterization of an extracellular protease from the fish pathogen Yersinia ruckeri and effect of culture conditions on production. Appl. Environ. Microbiol. 65: 3969-3075.
Seljelid, R. F., S. Bogwald, and A. Lundwall. 1981. Glycan stimulate of macrophage in vitro. Exp. Cell Res. 131: 121-129.
Shaw, D. H., M. J. Squires. 1984. O-antigen in a virulence strain of Aeromonas hydrophila. FEMS Microbiol. Lett. 24: 277-280.
Snieszko, S. F. 1958. Fish furunculosis. Fishery Leaflet 467. United States Fish and Wildlife Service, Washington, D.C. 4p.
Soffientino, B., T. Gwaltney, D. R. Nelson, J. L. Specker, M. Mauel, and M. Gomez-Choarri. 1999. Infectious necrotizing enteritis and mortality caused by Vibrio carchariae in summer flounder Paralichthys dentatus during intensive culture. Dis. Aquat. Org. 38: 201-210.
Song, Y. L. and G. H. Kou. 1981. Immune response of eel (Anguilla japonoca) against Aeromonas hydrophila and Edwardsiella anguilli-morteferrum (E. tarda) infection. Natural Science Communication Series. 3: 107-115.
Subsainghe, R. P., L. P. Jayasinghe, K. S. W. Balasuriya, and M. Kulathilake. 1990. Preliminary investigations into the bacterial and fungal pathogens associated with the ulcerative fish diseases syndrome in Sri Lanka, In” The second Asian fisheries Society, Manila. pp.655-657.
Thune, R. L., T. E. Grabam, L. M. Riddle, and R. L. Amborsk. 1982. Extracelular protease from Aeromonas hydrophila: partial and effects on age-0 channel catfish. Trans. Am. Fish. Soc. 111: 749-754.
Thune, R. L. and J. A. Plumb. 1982. Effect of delivery method and antigen preparation on the production of antibodies against Aeromonas salmonicida in channel catfish. Progressive Fish Culturist. 44: 53-54.
Thune, R. L., L. A. Stanley, and R. K. Copper. 1993. Pathologenesis of Gram-negative bacterial infections in warm-water fish. Annu. Rev. Fish. Dis. pp. 37-68.
Thune, R. L., J. P. Hawke, and R. J. Siebeling. 1991. Vibrio in the red swamp crawfish. J. Aquat. Anim. Heal. 3: 188-191.
Toranzo, A. E. and J. L. Barja. 1993. Virulence factors of bacteria pathogenic for coldwater fish. Ann. Rev. Fish Dis. 3: 5-36.
Torres, J. L., M. Shariff, and A. T. Law. 1990. Identification and virulence screening of Aeromonas spp. Isolated from healthy and epizootic ulcerative syndrome (EUS)-infected fish, In “ The Second Asian Fisheries Forum” (ed. By R. Hirano and I. Hanyu) The Asian Fisheries Society, Manila. pp.663-666.
Trust, T. J., M. Kostrzynska, L. Emody, and T. Wadstrom. 1993. High-affinity blinding of the basement membrane protein collagen type Ⅳ to the crystalline virulence surface protein array of Aeromonas salmonicida. Mol. Microbiol. 7: 593-600.
Trust, T. J., W. W. Kay, and E. E. Ishiguro. 1983. Cell surface hydrophobicity and macrophage association of Aeromonas salmonicida. Curr. Microbiol. 9: 315-318.
Van Duiln, C. 1981. Tuberculosis in fishes. Journal of Small Animal Practice. 22: 391-411.
Vipond, R., I. R. Bircknell, E. Durant, T. J. Bowden, A. E. Ellis, M. Smith, and S. MacIntyre. 1998. Defined delection mutants demostrate that the major secreted toxins are not essential for the virulence of Aeromonas salmonicida. Infect. Immun. 66: 1990-1998.
Wadstrom, T., A. Ljungh, and B. Wretlind. 1976. Enterotoxin, hemolysin and cytotoxic protein in Aeromonas hydrophila from human infections. Acta Pathol. Microbiol. Scand. Sect. B. 84: 112-114.
Wakabayashi, H. and S. Egusa. 1972. Characteristics of a Pseudomonas sp. From an epizootic of pond-cultured eels (Anguilla japonica). Bulletin of the Japanese Society of Scientific Fisheries. 38; 577-587.
Wagner, U., D. Hadge, B. K. Gudmundsdottir, K. Nold, and K. Drossler. 2001. Antibody response in salmonids against the 70 kDa serine protease of Aeromonas salmonicida studied by a monoclonal antibody-based ELISA. 2001. Vet. Immunol. Immunopathol. 28: 121-135.
Weisburg, W. G., S. M. Barns, D. A. Pelletier, and D. J. Lane. 1991. 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology. 173: 697-703.
Whitby, P. W., M. Landon, and G. Coleman. 1992. The cloning and nucleotide sequence of the serine protease gene (aspA) of Aeromonas salmonicida ssp. salmonicida. FEMS Microbiol. Lett. 99: 65-71.
Wood, E. M. and W. T. Yasutake. 1956. Histopathology of kidney disease in fish. American Jounal of Pathology. 32; 845-857.
Wretlind, B., L. Heden, and T. Wadstrom. 1973. Formation of extracellular hemolysin by Aeromonas hydrophila in relation to protease and staphylolytic enzyme. J. Gen. Microbiol. 78: 57-65.
Wretlind, B., R. Mollby, and Wadstrom, T. 1971. Separation of two haemolysins from Aeromonas hydrophila by isoelectric focusing. Infect. Immun. 4: 503-505.