親水性產器單胞菌（Aeromonas hydrophila）為溫水魚水產養殖中重要的致病菌，而防治疾病最安全、最有效率的方法即為疫苗的使用。為了研發更有效的疫苗，使用生物技術表現重要毒力因子以添加於傳統疫苗之中，可以增加疫苗的效果。親水性單胞菌有許多毒力因子已被發現，其中aerolysin具細胞破壞及溶解紅血球的功能，為重要的毒力因子之一。故本實驗中從親水性產氣單胞菌（Aeromonas hydrophila）中選殖出aerolysin之前趨蛋白 preproaerolysin 基因，得到含有1461個核酸的DNA片段，將其接入載體後以大腸桿菌表現系統大量表現，轉譯出487個氨基酸分子量約為54kDa的重組蛋白proaerolysin。重組蛋白經過trypsin活化切割後而具有溶血毒性的aerolysin。其在血液培養中能夠溶解紅血球而觀察到β-溶血的現象，在以細胞株之毒力測試試驗中亦可觀察到aerolysin對石斑魚細胞株具有導致細胞病變的毒性，在動物實驗中，aerolysin亦能夠造成點帶石斑魚的死亡。接下來再以基因工程的方式刪去此毒素毒性相關的不同區域之核酸序列，篩選得之失去毒性的重組蛋白，已不會產生溶血、細胞破壞及魚隻死亡的現象。使用此無毒性基因刪除的抗原蛋白具RPS 82~100％且具保護力之有效疫苗，較不活化疫苗之RPS 55~64％為優。

Aeromonas hydrophila is one of the most serious pathogen in Taiwan’s aquaculture. The safest and most efficient method of disease control is the preventive vaccine. Among vaccine development, the whole cell bactrine have been employed, but in some of the inactivated vaccine, it couldn’t induce sufficient immune response. The expressed virulence factor of pathogen by genetic engineering and added it to traditional vaccine has been shown to increase the efficacy. In this study, an important virulence factors of Aeromonas hydrophila, aerolysin, was selected as the antigen for vaccine development. The gene of aerolysin precursor -proaerolysin was isolated by PCR, and a 1461 base pair neucleotide was subcloned to an expression plasmid, and expression in E. coli. The inserted nuecleotide has translated a 487 amino acid, and with a molecular weight of 54 kDa. The protein r–proaerolysin was prepared from the inclusion body, trypsin digestion, and generated hemolytic and toxicity, on the blood agar test, the cell toxicity test and in animal text.
Deleting the toxic domain of proaerolysin by genetic engineering, three gene deleted proaerolysins have been obtain that have lost their cytotoxicity and hemolytic ability, that consequently lost the toxicity to fish. We have used three gene-deleted aerolysins as antigen in fish efficacy trail, and have found they could induce protective immunity, with RPS ranged from 82-100％ v.s the inactived whole bacterial vaccine the 55-64％.

Abrami L., Fivaz M., Glauser P.E., Sugimoto N., Zurzolo C., van der Goot FG. (2003) Sensitivity of Polarized Epithelial Cells to the Pore-Forming Toxin Aerolysin. Infect Immun. 71(2):739-46.
Agius C., Horne M.T., Ward P.D. (1983) Immunization of rainbow trout, Salmo
gairdneri Richardson, against vibriosis: comparison of an extract antigen kwith whole cell bacterin by oral and intraperitoneal routes. J Fish Dis. 6:129-34.
Alexander J.B., Bowers A., Shamsoom S.M.（1981） Hyperosmotic infiltration of bacteria into trout：route of entry and the fate of infiltrated bacteria. in Anderson DP, Hennessen W（eds）：Fish symposium：Serodiagnosis and Vaccines. Dev Biol Stand Basel Karger. 49：441-5
Altarriba M., Merino S., Gavín R., Canals R., Rabaan A., Shaw J.G., Tomás J.M. (2003) A polar flagella operon (flg) of Aeromonas hydrophila contains genes required for lateral flagella expression. Microb Pathog. 34(5):249-59.
Amend D.F., Fender D.C. (1976) Uptake of bovine serum albumin by rainbow trout from hyperosmotic infiltration: a model for vaccinating fish. Science. 192:793-4.
Anbarasu K., Thangakrishnan K., Aruna B.V.; Chandran M.R. (1998) Assessment of immune response in freshwater catfish (mystus vittatus bloch) to different bacterins of Aeromonas hydrophila. Indian J Exp Biol. 36(10):990-5 
Arkoosh, M. R., S. L. Kaatari. (1991) Development of immunological memory in rainbow trout(Oncorhynchus mykiss) Ⅰ. Am immunochemical and cellular analysis of the B cell response. Dev Comp Immunol. 15:279-93.
Asao T., Kinoshita Y., Kozaki S., Uemura T., Sakaguchi G. (1984) Purification and Some Properties of Aeromonas hydrophila Hemolysin. Infect Immun. 46(1):122-7.
Asha A., Nayak D.K., Shankar K.M., Mohan C.V. (2004) Antigen expression in biofilm cells of Aeromonas hydrophila employed in oral vaccination of fish. Fish Shellfish Immunol. 16(3):429-36.
Ballard J., Crabtree J., Roe B.A., Tweten R.K. (1995) The Primary Structure of Clostridium septicum Alpha-Toxin Exhibits Similarity with That of Aeromonas hydrophila Aerolysin. Infect Immun. 63(1):340-4.
Bernheimer A.W., Avigad L.S. (1974) Partial Characterization of Aerolysin, a Lytic Exotoxin from Aeromonas hydrophila. Infect Immun. 9(6):1016-21.
Bo J.N., Howard S.P. (1991) Mutagenesis and Isolation of Aeromonas hydrophila Genes Which Are Required for Extracellular Secretion. J Bacteriol. 173(3):1241-9.
Bootland L.M., Dobos P., Stevenson R.M.W.（1990）Fry age and size effects on
immersion immunization of brook trout, Salvelinus fontinalis Mitchell, againsy infectious pancreatic necrosis virus. J Fish Dis 13:113-25
Boulanger Y., Lallier R., Cousineau G. (1977) Isolation of enterotoxigenic Aeromonas from fish. Can J Microbiol. 23(9):1161-4.
Buckley J.T., Halasa L.N., MacIntyre S. (1982) Purification and partial characterisation of a bacterial phospholipid: cholesterol acyltransferase. J Biol Chem. 257(6):3320-5.
Buckley J.T., Wilmsen H.U., Lesieur C., Schulze A., Pattus F., Parker M.W., van der Goot FG. (1995) Protonation of Histidine- 132 Promotes Oligomerization of the Channel-Forming Toxin Aerolysin. Biochemistry. 34(50):16450-5.
Buckley J.T., Howard S.P. (1999) The Cytotoxic Enterotoxin of Aeromonas hydrophila Is Aerolysin. Infect Immun. 67(1):466-7
Buckley JT. (1992) Crossing three membranes. Channel formation by aerolysin. FEBS Lett. 307(1):30-3.
Bullock G.L., Chen P.K., Stuckey H.M. (1972) Studies of motile Aeromonads isolated from diseased warmwater and coldwater fishes.Page 21 in Abstracts of the Annual Meeting of the American Society for Microbiology, Philadelphia, Pennsylvania, 23-8 April 1972.
Busch R.A. (1982) "Enteric redmouth disease (Yersinia ruckeri) " in Anderson DP, Dorson M, Dubourget P (eds): Les antigènes des micro-organismes pathogènes des poissons, Collection Fondation Marcel Merieux,201-22.
Cascón A., Yugueros J., Temprano A., Sánchez M., Hernanz C., Luengo J.M., Naharro G. (2000). A major secreted elastase is essential for pathogenicity of Aeromonas hydrophila. Infect Immun. 68(6):3233-41.
Chakraborty T., Huhle B., Hof H., Bergbauer H., Goebel W. (1987) Marker exchange mutagenesis of the aerolysin determinant in Aeromonas hydrophila demonstrates the role of aerolysin in A. hydrophila-associated systemic infections. Infect Immun. 55(9):2274-80.
Colwell R.R., MacDonell M.R., De Ley J. (1986). Proposal to recognize the family Aeromonadaceae fam. nov. Int J Syst Bacteriol, 36:473–477.
de Kinkelin P. (1995) Vaccination in fish. Vet Res 26(3): 205-6.
Davidson, G.A., Ellis A.E., Secombes C.J. (1991) Cellular responses of leucocytes isolated from the gut of rainbow trout, Oncorhynchus mykiss (Walbaum). J Fish Dis. 14: 651-9.
Dixon P. (1997) Immunization with viral antigens: viral diseases of carp and catfish.Dev Biol Stand. 90: 221-32.
Dec C., Angel P., Baudin-Laurencin F. (1990) Effects of oral vaccination against vibriosis in turbot, Scophthalmus maximus L. and sea bass, Dicentrarchus labrax. J Fish Dis. 6:473-6
Dorson M. (1997) Vaccination trials of rainbow trout fry against infectious pancreatic necrosis. Bull of Int Epiz. 87:405-6
Duff D.C.B. (1942) The immunization of trout against bacterium salmonicida. J Immun 44:87-94.
Engelking H.M., Leong J.C., Follett J., Emmenegger E., Meyers T. (1991) Subunit vaccine to IHNV provides protection in sockeye salmon fry in Alaska. Amer Fish Soc Fish Health Newsletter. 19(2):4-5.
Espelid S., Løkken G.B., Bøgwald K. (1996) Effects of cortisol and stress on the immune system in Atlantic Salmon (Salmo salar L.). Fish Shellfish Immunol. 6:95-110
Espenes A., Press, CMcL, Dannevig B., Landsverk T. (1995) Immune-complex trapping in the splenic ellipsoids of rainbow trout (Oncorynchus mykiss). 282:41-8.
Evelyn T.P.T. (1997) A historical review of fish vaccinology. Dev Biol Stand. 90:3-12
Faktorovich K.A. (1969) Histological changes in the liver, kidneys, skin and brain of fish sick with red rot. Pages 83-101 in Infectious diseases of fish and their control. Division of Fisheries Research, Bureau of Sport Fisheries and Wildlife. Washington, D. C. Translated from the Russian by R. M. Howland.
Ferguson M.A., Low M.G., Cross G.A. (1985) Glycosyl-sn-1,2-dimyristylphosphatidylinositol is covalently linked to Trypanosoma brucei variant surface glycoprotein. J Biol Chem. 260(27):14547-55.
Ferguson M.A., Williams A.F. (1988) Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. Annu Rev Biochem. 57:285-320.
Ferguson M.R., Xu X.J., Houston C.W., Peterson J.W., Chopra A.K. (1995) Amino-acid residues involved in biological functions of the cytolytic enterotoxin from Aeromonas hydrophila. Gene. 156(1):79-83.
Fuentes R.J.M. and Perez H.J.A. (1998) Isolation of Aeromonas hydrophila in the rainbow trout (Oncorhynchus mykiss) Veterinaria Mexico. 29:117-119.
Garland W.J., Buckley J.T. (1998) The Cytolytic Toxin Aerolysin Must Aggregate To Disrupt Erythrocytes, and Aggregation Is Stimulated by Human Glycophorin. Infect Immun. 56(5):1249-53.
Glomset J.A., Gelb M.H., Farnsworth C.C. (1990) Prenyl proteins in eukaryotic cells: a new type of membrane anchor. Trends Biochem Sci. 15(4):139-42.
Goldes S.A., Ferguson H.W., Daoust P-Y, Moccia R.D. (1986) Phagocytosis of the inert suspended clay kaolin by the gills of rainbow trout salmo gairdneri. J Fish Dis. 9:147-52
González-Serrano C.J., Santos J.A., García-López M.L., Otero A. (2002) Virulence markers in Aeromonas hydrophila and Aeromonas veronii biovar sobria isolates from freshwater fish and from a diarrhoea case. J Appl Microbiol. 93(3):414-9.
Green M.J., Buckley J.T.(1990) Site-Directed Mutagenesis of the Hole-Forming Toxin Aerolysin: Studies on the Roles of Histidines in Receptor Binding and Oligomerization of the Monomer. Biochemistry. 29(8):2177-80.
Hantke K., Braun V. (1973) Covalent binding of lipid to protein. Diglyceride and amide-linked fatty acid at the N-terminal end of the murein-lipoprotein of the Escherichia coli outer membrane. Eur J Biochem. 34(2):284-96.
Hardie KR, Schulze A., Parker M.W., Buckly J.T.(1995) Vibrio spp. secrete proaerolysin as a folded dimmer without the need for disulphide bond formation. Mol Microbiol. 17(6):1035-44.
Moral H.C., Emilio Flaño del Castillo, Pilar López Fierro, Alberto Villena Cortés, Juan Anguita Castillo, Alberto Cascón Soriano, María Sánchez Salazar,1 Blanca Razquín Peralta, and Germán Naharro Carrasco (1998) Molecular characterization of the Aeromonas hydrophila aroA gene and potential use if an Auxotrophic aroA mutant as a live attenuated vaccine. Infect Immun. 66(5):1813-21.
Hart S., Wrathmell A.B., Harris J.E., Grayson T.H. (1988) Gut immunology in fish: a review. Dev Comp Immunol 12(3):453-80.
Hilton S., McCubbin W.D., Kay C.M., Buckley J.T. (1990) Purification and spectral study of a microbial fatty acyltransferase: activation by limited proteolysis. Biochemistry. 29(38):9072-8.
Holmes P., Niccolls L.M., Sartory D.P. (1996). The ecology of mesophilic
Aeromonas in the aquatic environment. In: Austin B et al., eds. The genus
Aeromonas. London, Wiley: 127–150.
Hong Y., Kang J.Y., Kim Y.U., Shin D.J., Choy H.E., Maeda Y., Kinoshita T. (2005) New mutant Chinese hamster ovary cell representing an unknown gene for attachment of glycosylphosphatidylinositol to protein. Biochem Biophys Res Commun. 335(4):1060-9.
Horne, M.T. (1997) Technical aspects of the administration of vaccines. Dev Biol Stand. 90:79-89.
Howard S.P., Buckley J.T. (1982) Membrane Glycoprotein Receptor and Hole-Forming Properties of a Cytolytic Protein Toxin. Biochemistry. 21(7):1662-7.
Howard SP, Buckley J.T. (1985) Activation of the Hole-Forming Toxin Aerolysin by Extracellular Processing. J Bacteriol. 163(1):336-40.
Howard S.P., Garland W.J., Green M.J., Buckley J.T. (1987) Nucleotide Sequence of the Gene for the Hole-Forming Toxin Aerolysin of Aeromonas hydrophila. J Bacteriol. 169(6):2869-71.
Huizinga H.W., Esch G.W., Hazen T.C. (1979) Histopathology of red-sore disease
(Aeromonas hydrophila) in naturally and experimentally infected largemouth bass Micropterus salmoides (Lacépède). J Fish Dis. 2:263-77.
Ikezawa H. (1999) Development from actions of bacterial phospholipases C on eucaryotic plasma membranes to molecular biology of GPI-anchored proteins. Yakugaku Zasshi. 119(7):472—94.
Ikezawa H. (2002) Glycosylphosphatidylinositol (GPI)-Anchored Proteins. Biol Pharm Bull. 25(4):409-17.
Janda J.M. (1991). Recent advances in the study of the taxonomy, pathogenicity, and infectious syndromes associated with the genus Aeromonas. Clin Microbiol Rev. 4:397–410.
Johnson K.A., Amend D.F. (1983) Efficacy of Vibrio anguillarum and Yersinia ruckeri bacterins applied by oral and anal intubation of salmonids. J Fish Dis 6:473-3.
Karunasagar I., Rosalind G. (1991) Immunological response of the Indian major carps to Aeromonas hydrophila vaccine. J Fish Dis. 14:413-7
Karunasagar I., Ali A., Otta S.K., Karunasagar I. (1997) Immunization with bacterial antigens: infections with motile aeromonads. Dev Biol Stand. 90:135-41.
Kawahara E., Kusuda R. (1988) "Location of Pasteurella piscicida antigen in tissue of yellowtail Seriola quinqueradiata vaccinated by immersion. Nippon Suissan Gakkaishi. 54:1101-5.
Kawai K., Kusuda R. (1985) Field testing of oral Vibrio anguillarum bacterin in pond-culturew ayu." Fish Pathol. 20(2/3):413-9.
Khushiramani R., Girisha S.K., Karunasagar I., Karunasagar I. (2007) Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish. Protein Expr Purif. 51(2):303-7
Khushiramani R., Girisha S.K., Karunasagar I., Karunasagar I. (2007) Protective efficacy of recombinant OmpTS protein of Aeromonas hydrophila in Indian major carp. Vaccine. 25(7):1157-8
Lamers C.H.J., de Haas M.J.H. (1985) Antigen localization in the lymphoid organs of carp (Cyprinus carpio). Cell Tissue Res. 242:491-8
Leblanc D., Mittal K.R., Olivier G., Lallier R. (1981) Serogrouping of motile Aeromonas species isolated from healthy and moribund fish. Appl Environ Microbiol. 42(1):56-60.
Lee K.-K., Ellis A.E. (1990) Glycerophospholipid:cholesterol acyltransferase complexed with lipopolysaccharide (LPS) is a major lethal exotoxin and cytolysin of Aeromonas salmonicida: LPS stabilizes and enhances the toxicity of the enzyme. J Bacteriol. 172:5382-93
Leong J.C., Anderson E., Bootland L., Chen L., Drolet B., Engelking H.M., C. Mason, D. Mourich, K. Johnson, G. Trobridge, J. Wirkkula. (1991) Biotechnical approaches to the development of salmonid fish vaccines, in Kimura T(ed):Proceedings of the OJI international Symposium on Salmonid Diseases, Sapporo, Hokkaido University Press:250-255
Leung K.Y., Stevenson R.M. (1988) Tn5-induced protease deficient strains of Aeromonas hydrophila with reduced virulence for fish. Infect Immun. 56(10):2639-44.
Lisanti M.P., Caras I.W., Gilbert T., Hanzel D., Rodriguez-Boulan E. (1990) Vectorial apical delivery and slow endocytosis of a glycolipid-anchored fusion protein in transfected MDCK cells. Proc Natl Acad Sci U S A. 87(19):7419-23
Loewy A.G., Santer U.V., Wieczorek M., Blodgett J.K., Jones S.W., Cheronis J.C. (1993). Purification and characterization of a novel zinc-proteinase of Aeromonas hydrophila. J Biol Chem. 268(12):9071-8.
Lumsden J.S., Ostland V.E., MacPhee D.D., Ferguson H.W. (1995) Production of gill-associated and serum antibody by rainbow trout (Oncorhynchus mykiss) following immersion immunization with acetone-killed Flavobacterium branchiophilum and the relationship to protection from experimental challenge. Fish Shellfish Immunol. 5:151-65
MacLean E., Ash R. (1986) The time-course of appearance and net accumulation of horseradish peroxidase (HRP) presented orally to juvenile caro Cyprinus carpio(L.). Comp Biochem Physiol. 84A(4):687-90.
Midtlyng P.J. (1997) Vaccinated fish welfare: protection versus side-effects. Dev Biol Stand 90:371-9.
Magarino B., Romalde J.L., Santos Y., Casal J.F., Barja J.L., Toranzo A.E. (1994) Vaccination trials on gilthead seabream (Sparus aurata) against Pasteurella piscicida. Aquaculture 120:201-8.
Mutoloki S., Brudeseth B., Reite O.B., Evensen O. (2006) The contribution of Aeromonas salmonicida extracellular products to the induction of inflammation in Atlantic salmon (Salmo salar L.) following vaccination with oil-based vaccines. Fish Shellfish Immunol. 20(1):1-11.
Nayak D.K., Asha A., Shankar K.M., Mohan C.V. (2004) Evaluation of biofilm of Aeromonas hydrophila for oral vaccination of Clarias batrachusda carnivore model. Fish Shellfish Immunol. 16(5):613-9.
Nishimura T., Sasaki H., Ushiyama M., Inoue K., Suzuki Y., Ikeya F., Tanaka M., Suzuki F.,Kohara M.,Arai M., Shima N., Sano T. (1985) A trial vaccination against rainbow trout fry with formalin kulled IHN virus. Fish Path. 20：435-43
Nybelin O. (1935) Uber agglutininbildung bei fischen. Z Immunitatsforsch Exp Ther. 84:74-9
Ogara W.O., Mbuthia P.G., Kaburia H.F.A., Sorum H., Kagunya D.K., Nduthu D.I., and Colquhoun D. (1998) Motile aeromonads associated with rainbow trout (Onchorhynchus mykiss) mortality in Kenya. Bulletin of the European Association of Fish Pathologists. 18:7-9.
Olafsen J.A., Hansen G.H. (1992) Intact antigen uptake in intestinal epithelial cells of marine fish larvae. J Fish Biol 40:141-56.
Ototake, M., Iwama G.K., Nakanishi T. (1986) The uptake of bovine serum albumin by the skin of bath immunized rainbow trout Oncorhynchus mykiss. Fish Shellfish Immunol. 6:321-33.
Paniagua C., Rivero O., Anguita J., Naharro G. (1990) Pathogenicity factors and virulence for rainbow trout (Salmo gairdneri) of motile Aeromonas spp. isolated from a river. J Clin Microbiol. 28(2):350-5.
Pemberton J.M., Kidd S.P., Schmidt R. (1997) Secreted enzymes of Aeromonas. FEMS Microbiol Lett. 152(1):1-10.
Pliszka F. (1939) Untersuchungen uber die agglutinine bei karpfen. Zentralbl Bakteriol Parasitenk, Infektionskr Hyg. Erste Abt: Medizin-hyg Baktertiol Virusforsch Parasitol. 143:262-4
Pliszka F. (1939) Weitere Untersuchungen uber Immunitatsreaktionen und uber Phagozytose bei Karpfen. Zentralbl Bakteriol Parasitenk, Infektionskr Hyg. Erste Abt: Medizin-hyg Baktertiol Virusforsch Parasitol. 143:451-60
Popoff M., Véron M. (1976) A taxonomic study of the Aeromonas hydrophila-Aeromonas punctata group. J Gen Microbiol. 94(1):11-22.
Quentel C. and Vigneulle M. (1997). Antigen uptake and immune responses after oral vaccination. Dev Biol Stand. 90: 69-78.
Rivero O., Anguita J., Paniagua C., Naharro G. (1990) Molecular cloning and characterization of an extracellular protease gene from Aeromonas hydrophila. J Bacteriol. 172(7):3905-8.
Rivero O., Anguita J., Mateos D., Paniagua C., Naharro G. (1991) Cloning and characterization of an extracellular temperature-labile serine protease gene from Aeromonas hydrophila. FEMS Microbiol Lett. 65(1):1-7.
Rombout J.H.W.M.A.J. Taverne-Thiele M.I. (1993a) The gut-associated lymphoid tissue (GALT) of carp (Cyprinus carpio L.): an immunocytochemical analysis. Dev Comp Immunol 17:55-66.
Rombout J.H.W.M., Block L.J., Lamers C.H.J., Egberts E. (1986) Immunization of carp (Cyprinus carpio) with a Vibrio anguillarum bacterium: indications for a common mucosal immune system. Dev Comp Immunol 10:341-51.
Rossjohn J, Buckley J.T., Hazes B., Murzin A.G., Read R.J., and Parker M.W. (1997) Aerolysin and pertussis toxin share a common receptor-binding domain. EMBO J. 16(12):3426-34
Rossjohn J., Raja S.M., Nelson K.L., Feil S.C., van der Goot FG, Parker M.W., Buckley J.T. (1998) Movement of a Loop in Domain 3 of Aerolysin Is Required for Channel Formation. Biochemistry. 37(2):741-6
Ruaugpan L., Kitao T., Yoshida T. (1986) Protective efficacy of Aeromonas hydrophila vaccines in nile tilapia. Vet Immunol Immunopathol. 12(1-4): 345-50
Sakai M., Aoki T., Kitao T., Rohovec J.S., Fryer J.L. (1984) Comparisons of the cellular immune response of fish vaccinated by immersion and injection of Vibrio anguillarum. Nippon Suisan Gakkaishi. 50:1187-92.
Santos N.M., Taverne-Thiele J.J., Barnes A.C., Muiswinkel W.B., Elli A.E., Rombout J.H. (2001) The gill is a major organ for antibody secreting cell production following direct immersion of sea bass (Dicentratchus labrax L.)in a Photobacterium damselae ssp. Piscida bacterin:an ontogenetic study. Fish Shellfish Immunol 11:65-74.
Schmidt M.F., Bracha M., Schlesinger M.J. (1979) Evidence for covalent attachment of fatty acids to Sindbis virus glycoproteins. Proc Natl Acad Sci U S A. 76(4):1687-91
Shieh H.S. (1987) Protection of atlantic salmon against motile Aeromonad septicaemia with Aeromonas hydrophila protease. Microbios Lett. 36(143-144): 133-138
Smith P.D. (1982) Analysis of the hyperosmotic and bath methods for fish vaccination:comparison of uptake of particulate and non-particelate antigens. Dev Comp Immunol. 2:81-186.
Smith W.W. (1940) Production of anti-bacterial agglutinins by carp and trout. Sov Exptl Med. 45:726-9
Song T., Toma C., Nakasone N., Iwanaga M. (2004) Aerolysin is activated by metalloprotease in Aeromonas veronii biovar sobria. J Med Microbiol. 53(Pt 6):477-82.
Tatner M.F. (1987) The quantitative relationship between vaccine dilution length of immersion time and antigen uptake using radiolabelled Aeromonas slmonicida bath in direct immersion experiments with rainbow trout Salmo gairdneri. Aquaculture. 62:173-86
Tebbit G.L., Goodrich T.D. (1982) "Vibriosis and the development of bacterins for its control" in Anderson DP, Dorson M, Dubourget P (eds): Les antigènes des micro-organismes pathogènes des poissons, Collection Fondation Marcel Merieux,201-22.
Thomas S.R., Trust T.J. (1995) A specific PulD homolog is required for the secretion of paracrystalline surface array subunits in Aeromonas hydrophila. J Bacteriol. 177(14):3932-9.
Thornton J., Howard S.P., Buckley J.T. (1988) Molecular cloning of a phospholipid-cholesterol acyltransferase from Aeromonas hydrophila. Sequence homologies with lecithin-cholesterol acyltransferase and other lipases. Biochim Biophys Acta. 959(2):153-9
Tucker A.D., Parker M.W., Tsernoglou D., Buckley J.T. (1990) Crystallization of a Proform of Aerolysin, a Hole-forming Toxin from Aeromonas hydrophila. J Mol Biol. 212(4):561-2.
van der Goot FG, Hardie K.R., Parker M.W., Buckley J.T. (1994) The C-terminal peptide produced upon proteolytic activation of the cytolytic toxin aerolysin is not involved in channel formation. J. Biol. Chem. 269: 30496-501
van Muiswinkel W.B., Wiegertjes G.F. (1997) Immune responses after injection vaccination of fish. Dev Biol Stand. 90: 55-7.
Vivas J., Razquin B., López-Fierro P., Villena A.J. (2005) Modulation of the immune response to an Aeromonas hydrophila aroA live vaccine in rainbow trout: effect of culture media on the humoral immune response and complement consumption. Fish Shellfish Immunol. 18(3):223-33.
Vivas J., Riaño J., Carracedo B., Razquin B.E., López-Fierro P., Naharro G., Villena A.J. (2004) The auxotrophic aroA mutant of Aeromonas hydrophila as a live attenuated vaccine against A. salmonicida infections in rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immunol. 16(2):193-206.
Wang G., Clark C.G., Liu C., Pucknell C., Munro C.K., Kruk T.M., Caldeira R., Woodward D.L., Rodgers F.G. (2003) Detection and Characterization of the Hemolysin Genes in Aeromonas hydrophila and Aeromonas sobria by Multiplex PCR. J Clin Microbiol. 41(3):1048-54.
Whittington R.J., Munday B.L., Akhlaghi M., Reddacliff G.L., Carson J. (1994) Humoral and peritoneal cell responses of rainbow trout (Oncorhynchus myjiss) to ovalbumin, Vibrio anguillarum and Freund’s completet adjuvant following intrapperitoneal and bath immunization. Fish Shellfish Immunol. 4:475-88
Wong C.Y., Heuzenroeder M.W., Flower R.L. (1998) Inactivation of two haemolytic toxin genes in Aeromonas hydrophila attenuates virulence in a suckling mouse model. Microbiology. 144 ( Pt 2):291-8
Wong K.R., Buckley J.T. (1991) Site-directed Mutagenesis of a Single Tryptophan near the Middle of the Channel-forming Toxin Aerolysin Inhibits Its Transfer across the Outer Membrane of Aeromonas samonicida. J Biol Chem. 266(22):14451-6.
Wong K.R., McLean D.M., Buckley J.T. (1990) Cloned aerolysin of Aeromonas hydrophila is exported by a wild-type marine Vibrio strain but remains periplasmic in pleiotropic export mutants. J Bacteriol. 172(1):372-6.
Wretlind B., Möllby R., Wadström T. (1971) Separation of Two Hemolysins from Aeromonas hydrophila by Isoelectric Focusing. Infect Immun. 4(4):503-5.
Yambot A.V., Inglis V. (1994) Aeromonas hydrophila isolated from Nile tilapia
(Oreochromis niloticus L.) with "Eye Disease". International Symposium on Aquatic Animal Health, Seattle, WA (USA), 4 - 8 September 1994. University of California, School of Veterinary Medicine, Davis, CA. p.103.
林青丘(2002)，魚用生物包埋型口服疫苗製備與有效性測試，國立成功大學生物科技研究所碩士論文
黃卓男(2001)，香魚分離親水性產氣單胞菌Aeromonas hydrophila重組aerolysin 之表現及特性，國立台灣大學獸醫學研究所碩士論文