現今鰻魚(Anguilla japonica)的疾病，大部分是用抗生素來治療，但是抗生素的使用並非只抑制一種細菌，可能同時將一些有利於鰻魚的細菌也一起殺死，使得鰻魚不易存活，同時也造成抗藥性因子散佈和累積，對於日後施藥的成效和公共衛生也將有重大之影響。目前的研究趨勢，是找出具有競爭性且不會致病的細菌，來抑制同一生態中的病原菌。本實驗以鰻魚潰瘍病致病菌愛德華氏菌(Edwardsiella tarda)為對象，從鰻魚養殖環境中，篩選出可以抑制愛德華氏菌的細菌，供日後發展生物製劑之用。本研究共測試了2412株菌，得到8株菌有抑制E. tarda的能力，其有效抑制E. tarda物質分子量皆小於12,000 daltons。利用16S rDNA序列分析方法，得其中1株為Pseudomonas aeruginosa相似度為99.86%、1株為Bacillus cereus相似度為99.93%、4株為Bacillus sphaericus相似度為98.48~98.62%之間、2株為Bacillus laterosporus相似度為98.76%和99.16%，鑑定菌種可至種的階段，也做了一些輔助性生理生化測試加以佐證。競爭測試P. aeruginosa對愛得華氏菌拮抗效果很好，其餘三群屬於Bacillus屬，競爭效果就不理想。如此，但文獻中指出Pseudomonas和Bacillus屬，是維護或控制養殖環境水質效果很好的菌屬，值得進一步研究。

草蝦(Penaeus monodon)養殖，在台灣曾風光一段時間，但民國77年發生大量死亡之後，至目前卻一蹶不振，因此研究者開始探討草蝦死亡的原因並加以解決，例如：提高草蝦本身防預能力，了解並對付病原，而養殖生態環境也是不可忽略的重要一環。另一個問題，即養殖草蝦抽取地下水，造成地層下陷的問題，少用水的養殖方式則成為現代養殖的新趨勢。本實驗為一封閉式，以優勢菌來控制養殖草蝦水質，研究池水中，可培養細菌之種類，可供日後養殖之參考。以16S rDNA序列分析方法來鑑定，共約1,500 base數，也以BIOLOG做為鑑定的輔助，但兩種方法結果相差很大。以16S rDNA序列分析，得9個族群，分別為Vibrio vulnificus相似度為99.82~100.00%、Vibrio mediterranei相似度為97.26%、Pseudoalteromonas相似度為98.83%、Porphyrobacter相似度為99.42%、Flavobacterium相似度為89.99和90.45%、Rhodthermus相似度為81.26%和3群分類地位較不明確族群，相似度分別為89.80~90.67%、98.26%和97.74%與97.21%和97.90%。鑑定至種階段者有V. vulnificus、Porphyrobacter、Pseudoalteromonas和V. mediterranei。和文獻做比較，有幾群相似，例如：Vibrio和Flavobacterium屬等，而其餘則不同，可能原因是養殖環境不同或分類方法不同所造成。

Presently, most bacterial diseases of eel (Anguilla japonica) are controlled by antibiotics. However, antibiotics not only kill the bacterial pathogens but also kill those bacteria which might be beneficial to eels. In the meantime, application of antibiotics may result in spreading and accumulation of the resistance genes which may in turn lower the efficacy the antibiotics in the future and may threat public health. The recent trend to such problems is to screen non-pathogenic bacteria which are competitive to the pathogenic bacteria in the same environments. The eel pathogen, Edwardsiella tarda, was chosen as the target in this study. Bacterial strains were isolated from different eel ponds and tested for the ability to inhibit the growth of E. tarda. Of 2,412 strains tested, eight of them showed the inhibition capability. The molecular weights of the bioactive ingredients are all smaller than 12,000 daltons indicating they are not protein in nature. One of the strains is Bacillus cereus, four of the strains are Bacillus sphaericus, two of the strains are Bacillus laterosporus, and one of the strains of identified as Pseudomonas areuginosa competed extremely well with E. tarda. These antagonistic bacteria may have the potential of becoming as bio-control agents.Tiger shrimp (Penaeus monodon) is an important agricultural product in Taiwan. The over all production peaked in 1988, since then the outbreak of viral infection has caused the shrimp aquaculture a heavy damage. The current production is merely 1/10 of the peak. Many solutions were proposed to solve the problem, such as: increase the immunity of the shrimp, study pumping of the underground water has caused serious land subsidence in the coastal areas. Therefore, conservation of water is the trend of current aquaculture. In this study, culturable bacteria were isolated from a closed tiger shrimp pond. The taxonomy of the bacteria was based on 16S rDNA sequence phylogeny. Roughly 8 groups (genera) of bacteria were identified, including: Vibrio, Pseudoalteromonas, Porphyrobacter, Flavobacterium, Rhodthermus and three uncertain genera.

壹.鰻魚病原菌Edwardsiella tarda拮抗菌株研究……………………….1
摘要…………………………………………………………………….2
Abstract…………………………………………………………………3
前言…………………………………………………………………….4
材料與方法…………………………………………………………….9
結果……………………………………………………………………19
討論……………………………………………………………………24
參考文獻………………………………………………………………30

貳.草蝦池水可培養菌株研究…………………………………………….57
摘要……………………………………………………………………58
Abstract…………………………………………………………………59
前言……………………………………………………………………60
材料與方法……………………………………………………………64
結果……………………………………………………………………71
討論……………………………………………………………………78
參考文獻………………………………………………………………82





表 目 錄

表次 頁次
表一 E. tarda拮抗菌與其最相似菌株比較……………………………38
表二 拮抗菌P. aeruginsa、B. cereus、B. sphaericus和B. laterosporus
型態、生理、生化測驗………………..…………………………39
表三 草蝦池水菌相中各族群之數量及BIOLOG鑑定結果………….87
表四 草蝦池水中可培養菌株與其最相似菌株比較…………………..88
表五 草蝦池水中可培養菌之同族群間相似度比較…………………..89
表六 草蝦池水可培養菌之相似度……………………………………..90
表七 草蝦池水可培養菌之演化距離…………………………………..91

















圖 目 錄

圖次 頁次
圖一 E. tarda拮抗菌之16S rDNA序列………………………………….40
圖二 E. tarda和P. aeruginosa (E1)競爭(拮抗)實驗之菌數變化曲線…..49
圖三 E. tarda和P. aeruginosa競爭實驗之總菌數百分比變化…………50
圖四 E. tarda和B. cereus (E2)競爭(拮抗)實驗之菌數變化曲線……….51
圖五 E. tarda和B. cereus競爭實驗之總菌數百分比變化……………...52
圖六 E. tarda和B. sphaericus (E4)競爭(拮抗)實驗之菌數變化曲線…..53
圖七 E. tarda和B. sphaericus競爭實驗之總菌數百分比變化…………54
圖八 E. tarda和B. laterosporus (E7)競爭(拮抗)實驗之菌數變化曲線…55
圖九 E. tarda和B. laterosporus競爭實驗之總菌數百分比變化……….56
圖十 草蝦池可培養菌之16S rDNA序列…………………………….…..92
圖十一 草蝦池水之可培養菌16S rDNA rooted phylogenetic tree圖……...105
圖十二 草蝦池水可培養菌16S rDNA unrooted phylogenetic tree圖……...106

Al-Hadhrami, M. N., H. M. Lappin-Scitt, and P. J. Fisher. 1996. Effects of the addition of organic carbon sources on bacterial respiration and n-alkane biodegradation of Omani crude oil. Marine Pollution Bulletin 32:351-357.

Altschul, S. F., W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. 1990. Basic local alignment search tool. Journal of Molecular Biology 215:403-410.

Amandi, A., S. F. Hiu, J. S. Rohovec, and J. L. Fryer. 1982. Isolation and characterization of Edwardsiella tarda from fall chinook salmon (Oncorhynchus tshawytscha). Applied and Environmental Microbiology 43:1380-1384.

Assche, J. V. 1991. Edwardsiella tarda infection in a puppy with possible parvovirus infection. The Veterinary Record 23:475-476.

Atlas, R. M., and R. Bartha. 1987. Micriobial ecology fundamentals and applications (2nded.). The Benjamin and Cummings Publi. Co. Inc. Menlo Park, California.

Aziz, N. H., M. Z. El-Fouly, A. A. El-Essawy, and M. A. Khalaf. 1997. Influence of bean seedling root exudates on the rhizosphere colonization by Trichoderma lignorum for the control of Rhizoctonia solani. Botanical Bulletin of Academia Sinica 38:33-39.

Aznar, R., W. Ludwig, R. I. Amann, and K. H. Schleifer. 1994. Sequence determination of rRNA genes of pathogenic Vibrio species and whole-cell identification of Vibrio vulnificus with rRNA-targeted oligonucleotide probes. International Journal of Systematic Bacteriology 44:330-337.

Baumann, P., M. A. Clark, L. Baumann, and A. H. Broadwell. 1991. Bacillus sphaericus as a mosquito pathogen: properties of the organism and its toxins. Microbiological Reviews 55:425-436.

Baya, A. M., J. L. Romalde, D. E. Green, R. B. Navarro, J. Evans, E. B. May, and A. E. Toranzo. 1997. Edwardsiellosis in wild striped bass from the Chesapeake Bay. Journal Wildlife Diseases 33:517-525.

Berner, R., F. Heinen, K. Pelz, V. V. Velthoven, M. Sauer, and R. Korinthenberg. 1997. Ventricular shunt infection and meningitis due to Bacillus cereus. Neuropediatrics 28:333-334.

Block, C. S., M. L. Levy, and V. U. Fritz. 1978. Bacillus cereus endocarditis: a case report. South African Medical Journal 53:556-557.

Burdett, I. D. J. 1988. Structure, growth, and division of the Bacillus subtilis cell surface. Canadian Journal of Microbiology 34:373-380.

Canyon, D. V., and J. L. K. Hii. 1997. The gecko: an environmentally friendly biological agent for mosquito control. Medical and Veterinary Entomology 11:319-323.

Charles, J.-F., C. Nielsen-LeRoux, and A. Delecluse. 1996. Bacillus sphaericus toxins: molecular biology and mode of action. Annual Review of Entomology 41:451-472.

Chen, J.-D., S.-Y. Lai, and S.-L. Huang. 1996. Molecular cloning, characterization, and sequencing of the hemolysin gene from Edwardsiella tarda. Archives of Microbiology 165:9-17.

Chernin, L., A. Brandis, Z. Ismailov, and I. Chet. 1996. Pyrrolnitrin production by an Enterobacter agglomerans strain with a broad spectrum of antagonistic activity towards fungal and bacterial phytopathogens. Current Microbiology 32:208-212.

Costa, A. B., K. Kanai, and K. Yoshikoshi. 1998. Serological characterization of atypical strains of Edwardsiella tarda isolated from sea breams. Fish Pathology 33:265-274.

Curtis, J. R., A. J. Wing, and J. C. Coleman. 1967. Bacillus cereus bacteraemia: a complication of intermittent haemodialysis. The Lancet 1:136-138.

Depaola, A., J. T. Peeler, and G. E. Rodrick. 1995. Effect of oxytetracycline-medicated feed on antibiotic resistance of Gram-negative bacteria in catfish ponds. Applied and Environmental Microbiology 61:2335-2340.

Favret, M. E., and A. A. Yousten. 1985. Insecticidal activity of Bacillus laterosporus. Journal of Invertebrate Pathology 45:195-203.

Grenier, D. 1996. Antagonistic effect of oral bacteria towards Treponema denticola. Journal of Clinical Microbiology 34:1249-1252.

Grondona, I., R. Hermosa, M. Tejada, M. D. Gomis, P. F. Mateos, P. D. Bridge, E. Monte, and I. Garcia-acha. 1997. Physiological and biochemical characterization of Trichoderma harzianum, a biological control agent against soilborne fungal plant pathogens. Applied and Environmental Microbiology 63: 3189-3198.

Groschel, D., M. A. Burgess, and G. P. Bodey. 1976. Gas gangrene-like infection with Bacillus cereus in a lymphoma patient. Cancer 37:988-991.

Gutierrez, M. A., T. Miyazaki, H. Hatta, and M. Kim. 1993. Protective properties of egg yolk IgY containing anti-Edwardsiella tarda antibody against paracolo disease in the Japanese eel, Anguilla japonica Temminck and Schlegel. Journal of Fish Diseases 16:113-122.

Harwood, C. R. 1989. Introduction to the biotechnology of Bacillus. p.1-4. In: Harwood, C. R. (ed.). Bacillus. Plenum Press, New York.

Hossain, Md. A., M. Furuichi, and Y. Yone. 1988. Microbial treatment of liquid from mackerel waste juice. Nippon Suian Gakkaishi. 54:2113-2115.


Hung, H.-W., C.-F. Lo, C.-C. Tseng, and G.-H. Kou. 1997a. Antibody production in Japanese eels, Anguilla japonica Temminck and Schlegel. Journal of Fish Diseases 20:195-200.

Hung, H.-W., C.-F. Lo, C.-C. Tseng, and G.-H. Kou. 1997b. Antibody response of glass eels, Anguilla japonica Temminck and Schlegel, to Pleistophora anguillarum Hoshina (microspora) infection. Journal of Fish Diseases 20:237-239.

Itou, T., T. Iida, and H. Kawatsu. 1997. The importance of hydrogen peroxide in phagocytic bactericidal activity of Japanese eel neutrophils. Fish Pathology 32:121-125.

Janda, J. M., and S. L. Abbott. 1993. Infection Associated with the gene Edwardsiella : the role of Edwardsiella tarda in humun disease. Clinical Infectious Diseases 17:742-748.

Kalfon, A., J.-F. Charles, C. Bourgouin, and H. D. Barjac. 1984. Sporulation of Bacillus sphaericus 2297: an electron microscope study of crystal-like inclusion biogenesis and toxicity to mosquito larvae. Journal of General Microbiology 130:893-900.

Kita-Tsukamoto, K., H. Oyaizu, K. nanba, and U. Simidu. 1993. Phylogenetic relationships of marine bacteria, mainly member of the family Vibrionaceae, determined on the basis of 16S rRNA sequences. International Journal of Systematic Bactriology 43:8-19.

Kokubo, T., T. Iida, and H. Wakabayashi. 1990. Production of siderophore by Edwardsiella tarda. Fish Pathology 25:237-241.

Komukai-Nakamura, S., K. Sugiura, Y. Yamauchi-Inomata, H. Toki, K. Venkateswaran, S. Yamamoto, H. Tanaka, and S. Harayama. 1996. Construction of bacterial consortia that degrade Arabian light crude oil. Journal of Fermentation and Bioenginering 82:570-574.

Krieg, N. R., and J. G. Holt. 1984. Bergey’s manual of systematic bacteriology volume l. p.164-165, and p.486-489. Williums & wilkins, USA.

Kusuda, R., and K. Kawai. 1998. Bacterial diseases of cultured marine fish in Japan. Fish Pathology 33:221-227.

Kusuda, R., K. Ono, and F. Salati. 1988. Changes in agglutinating antibody titers and serum proteins of immunized eels. Diseases of Aquatic Organisms 5:87-92.

Lovley, D. R., D. F. Dwyer, and M. J. Klug. 1982. Kinetic analysis of competition between sulfate reducers and methanogens for hydrogen in sediments. Applied and Environmental Microbiology 43:1373-1379.

Lund, B. M. 1990. Foodborne illness. The Lancet 336:982-986.

Luz, C., I. G. Silva, C. M. T. Cordeiro, and M. S. Tigano. 1998. Beauveria bassiana (hyphomycetes) as a possible agent for biological control of Chagas disease vectors. Journal of Medical Entomology 35:977-979.

Mahler, H., A. Pasi, J. M. Kramer, P. Schulte, A. C. Scoging, W. Bar, and S. Krahenbuhl. 1997. Fuminant liver failure in association with the emetic toxin of Bacillus cereus. New England Journal of Medicine 336:1142-1148.

Mano, N., T. Inui, D. Arai, H. Hirose, and Y. Deguchi. 1996. Immune response in the skin of eel against Cytophaga columnaris. Fish Pathology 31:65-70.

Middlebrook, J. L., and R. B. Dorland. 1984. Bacterial toxins: cellular mechanisms of action. Microbiological Reviews 48:199-221.

Miyazaki, T., M. A. Gutierrez, and S. Tanaka. 1992. Experimental infection of Edwardsiellosis in the Japanese eel. Gyobyo Kenkyu 27:39-47.

Nakamura, S., Y. Gohya, J. N. Losso, S. Nakai, and A. kato. 1996. Protective effect of lysozyme-galactomannan or lysozyme-palmitic acid conjugates against Edwardsiella tarda infection in carp, Cyprinus carpio L. FEBS Letters 383: 251-254.

Nordera, P., and G. Menestrina. 1998. Proteolytic cleavage of Pseudomonas aeruginosa extoxin A in the pesence of lipid bilayers of different composition. FEBS Letters 421:268-272.

Nougayrede, PH., A. Vuillaume, M. Vigneulle, B. Faivre, S. Luengo, and J. Delprat. 1994. First isolation of Edwardsiella tarda from diseased turbot (Scophthalmus maximus) reared in a sea farm in the bay of Biscay. Bulletin of the European Association of Fish Pathologists 14:128-129.

Oei, C., J. Hindley, and C. Berry. 1992. Binding of purified Bacillus sphaericus binary toxin and its deletion derivatives to Culex quinquefasciatus gut: elucidation of functional binding domains. Journal of General Microbiology 138:1515-1526.

Orlova, M. V., T. A. Smirnova, L. A. Ganushina, V. Y. Yacubovich, and R. R. Azizbekyan. 1998. Insecticidal activity of Bacillus laterosporus. Applied and Environmental Microbiology 64:2723-2725.

Owens, D. R., S. L. Nelson, and J. B. Addison. 1974. Isolation of Edwardsiella tarda from swine. Applied Microbiology 27:703-705.

Park, K. H., and H. D. Jeong. 1996. Enhanced resistance against Edwardsiella tarda infection in tilapia (Oreochromis niloticus) by administration of protein-bound polysaccharide. Aquaculture 143:135-143.

Park, S.-W., and H. Wakabayashi. 1992. Comparison of pronephric and peripheral blood neutrophils of eel, Anguilla japonica, in phagocytic activity. Gyobyo Kenkyu 27:149-152.

Payne, J. M., and E. W. Davidson. 1984. Insecticidal activity of the crystalline parasporal inclusions and other components of the Bacillus sphaericus 1593 spore complex. Journal of Invertebrate Pathology 43:383-388.

Pearson, H. E. 1970. Human infections caused by organisms of the Bacillus species. American Journal of Clinical Pathology 53:506-515.

Peruski Jr, L. F., and A. H. Peruski. 1997. Analysis of sequence data, p.206-254. In: L. F. Peruski Jr, and A. H. Peruski.(ed.). The internet and the new biology. American Society for Microbiology. Washington, D. C.

Pien, F. D., and M. T. Jackson. 1995. Tuboovarian abscess caused by Edwardsiella tarda. American Journal of Obstetrics and Gynecology 173:964-965.

Pier, G. B., G. J. Small, and H. B. Warren. 1990. Protection against mucoid Pseudomonas aeruginosa in rodent models of endobronchial infections. Science 249:537-540.

Poncet, S., C. Bernard, E. Dervyn, J. Cayley, A. Klier, and G. Rapoport. 1997. Improvement of Bacillus sphaericus toxicity against dipteran larvae by integration, via homologous recombination, of the CryllA toxin gene from Bacillus thuringiensis subsp. israelensis. Applied and Environmental Microbiology 63:4413-4420.

Porter, A. G. 1996. Mosquitocidal toxins, genes and bacteria: the hit squad. Parasitology Today 12:175-179.

Porter, A. G., E. W. Davidson, and J.-W. Liu. 1993. Mosquitocidal toxins of Bacilli and their genetic manipulation for effective biological control of mosquitoes. Microbiological Reviews 57:838-861.

Priest F. G., L. Ebdrup, V. Zahner, and P. E. Carter. 1997. Distribution and characterization of mosquitocidal toxin genes in some strains of Bacillus sphaericus. Applied and Environmental Microbiology 63:1195-1198.

Priest, F. G. 1989. Isollation and identification of aerobic endospore-forming bacteria. p.27-36. In: Harwood, C. R. (ed.). Bacillus. Plenum Press, New York.

Rahme, L. G., E. J. Stevens, S. F. Wolfort, J. Shao, R. G. Tompkins, and F. M. Ausubel. 1995. Common virulence factors for bacterial pathogenicity in plants and animals. Science 268:1899-1902.

Reithinger, R., C. R. Davies, H. Cadena, and B. Alexander. 1997. Evaluation of the fungus Beauveria bassiana as a potential biological control agent against phlebotomine sand flies in Colombian coffee plantations. Journal of Invertebrate Pathology 70:131-135.

Rivers, D. B., C. N. Vann, H. L. Zimmack, and D. H. Dean. 1991. Mosquitocidal activity of Bacillus laterosporus. Journal of Invertebrate Pathology 58:444-447.

Ruangpan, L., P. Na-anan, and S. Direkbusarakom. 1998. Inhibitory effect of Vibrio alginolyticus on the growth of V. hareyi. Fish Pathology 33:293-296.

Sakai, M., S. Atsuta, and M. Kobayashi. 1994. Survival of fish pathogen Edwardsiella tarda in sea water and fresh water. Bulletin of the European Association of Fish Pathologists 14:188-190.

Sambrook, J. E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning: a laboratory manual (2nded.). Cold Spring Harbor Laboratory Press. New York.

Schoenfeld, P. S., J. W. Myers, L. Myers, and J. C. Larocque. 1995. Suppression of cell-mediated immunity in hypothyroidism. Southern Medical Journal 88:347-349.

Slepecky, R. A. 1992. What is Bacillus. p.1-22. In: Doi, R. H., and M. M. Gloughlin (ed.). Biology of Bacillus. Butterworth Heinemann, USA.

Solny, M. N., G. R. Failing, and J. S. Borges. 1977. Bacillus cereus osteomyelitis. Archives of Internal Medicine 137:401-402.

Sonnenwirth, A. C., and B. A. Kallus. 1968. Meningitis due to Edwardsiella tarda. The American Journal of Clinical Pathology 49:92-95.

Stackebrandt, E., and M. Goodfellw. 1991. Nucleic acid techniques in bacterial systematics. p.72-73. John Wiley&Sons.

Strauss, E. J., N. Ghori, and S. Falkow. 1997. An Edwardsiella tarda strain containing a mutaion in a gene with homology to shlB and hpmB is defective for entry into epithelial cells in culture. Infection and Immunity 65:3924-3932.

Sugahara, I., L. R. Berger, T. Kimura, and K. Hayashi. 1988. Effects of temperature and pressure on the lytic and autolytic activites of Bacillus sp. V37 isolated from coastal regions. Nippon Suisan Gakkaishi 54:861-867.

Sugahara, I., T. Kimura, Y. Nakagawa, H. Kojima, M. Usuda, and M. Tanaka. 1990. Lytic enzyme-producing bacteria in marine bottom sediments. Nippon Suisan Gakkaishi 56:511-518.

Suprapto, H., T. Nakai, and K. Muroga. 1995. Toxicity of extracellular products and intracellular components of Edwardsiella tarda in the Japanese eel and flounder. Journal of Aquatic Animal Health 7:292-297.

Suprapto, H., T. Hara, T. Nakoi, and K. Muroga. 1996. Purification of a lethal toxin of Edwardsiella tarda. Fish Pathology 31:203-207.

Swannell, R. P. J., K. Lee, and M. Mcdonagh. 1996. Field evaluations of marine oil-spill bioremediation. Microbiological Reviews 60:342-365.

Terranova, W., and P. A. Blake. 1978. Bacillus cereus food poisoning. New England Journal of Medicine 298:143-144.

Thompson, J. D., D. G. Higgins, and T. J. Gibson. 1994. Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Reserch 22:4673-4680.

Tison, D., M. Nishibuchi, J. D. Greenwood, and R. J. Seidler. 1982. Vibrio vulnificus biogroup 2: new biogroup pgthogenic for eels. Applied and Environmental Microbiology 44:640-646.

Turnbull, P. C. B., K. Jorgensen, J. M. Kramer, R. J. Gilbert, and J. M. Parry. 1979. Severe clinical conditions associated with Bacillus cereus and the apparent involvement of exotoxins. Journal of Clinical Pathology 32:289-293.

Vandepitte, J., P. Lemmens, and L. D. Swert. 1983. Human Edwardsiellosis traced to ornamental fish. Journal of Clinical Microbiology 17:165-167.

Vogel, B. F., K. Jorgensen, H. Christensen, J. E. Olsen, and L. Gram. 1997. Differentiation of Shewanella putrefaciens and Shewanella alga on the basis of whole-cell protein profiles, ribotyping, phenotypoic characterization, and 16S rRNA gene sequence analysis. Applied and Environmental Microbiology 63:2189-2199.

Waltman, W. D., and E. B. Shotts. 1986. Antimicrobial susceptibility of Edwardsiella tarda from the united states and Taiwan. Veterinary Microbiology 12:277-282.

Zighelboim, J., T. W. Williams, M. W. Bradshaw, and R. L. Harris. 1992. Successful medical management of a patient with multiple hepatic abscesses due to Edwardsiella tarda. Clinical Infectious Diseases 14:117-120.

江柏輝. 1998. 以微生物製劑淨化養殖水質之研究. 國立臺灣海洋大學水產養殖研究所. 碩士論文.

吳金洌, 趙偉真和劉國鈞. 1987. 感染潰瘍病細菌Edwardsiella tarda之ψET-1噬菌體之生物性質研究. 魚病研究論文集(三)病毒及藥物專輯, 農委會漁業特刊. 第十二號:24-33.

吳金洌和趙偉真. 1987. 感染潰瘍病細菌Edwardsiella tarda之新噬菌體株ψET-1之研究. 魚病研究論文集(三)病毒及藥物專輯, 農委會漁業特刊. 第十二號:9-18.

吳俊德. 1997. 出血性紅斑病弧菌（Vibrio anguillarum）及腸炎弧菌（Vibrio parahaemolyticus）噬菌體之研究. 國立中山大學海洋資源研究所. 碩士論文.

林義雄, 鍾虎雲和郭光雄. 1989. 台灣水產養殖魚類常見的細菌性病害. 中國水產月刊 433:6-12.

徐玉標, 江漢全和林承民. 1986. 養殖用水之現況調查研究. 水產養殖研究報告彙集(一), 農委會漁業特刊. 第九號:125-137.

張金豐. 1977. 鰻魚池生態之研究. 私立中國文化學院海洋研究所. 碩士論文.

郭玉梅. 1999. 油污分解菌之研究. 國立中山大學海洋資源研究所. 碩士論文.

陳宏遠. 1978a. 鰻魚抗藥細菌之抗藥性. 國立臺灣大學海洋研究所. 碩士論文.

陳昭德. 1978b. 養殖鰻細菌分佈之研究. 國立臺灣大學動物學研究所. 碩士論文.

曾萬年. 1996. 鰻線漁獲量的變動及其原因. 中國水產月刊 446:5-11.

黃旭田和涂堅. 1992. 魚類細菌性疾病之分離與診斷. p.92-95. 台灣養豬科學研究所出版.

劉文御. 1995. 台灣養殖蝦池微生物生態學研究. 國立臺灣大學動物學研究所. 博士論文.

劉朝鑫和王建雄. 1986. 魚類病原菌抗藥性之研究-II, 分佈於養殖環境中之Edwardsiella tarda的抗藥性. 魚病研究專集(八), 農委會漁業特刊. 第八號:56-67.

羅仲邑. 1997. 噬菌體控制鰻魚(Anguilla japonica)病原菌Aeromonas hydrophila, Edwardsiella tarda之研究. 國立中山大學海洋資源研究所. 碩士論文.Altschul, S. F., W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. 1990. Basic local alignment search tool. Journal of Molecular Biology 215:403-410.

Andresson, O. S., and O. H. Fridjonsson. 1994. The sequence of the single 16S rRNA gene of the thermophilic eubacterium Rhodothermus marinus reveals a distant relationship to the group containing Flexibacter, Bacteroides, and Cytophaga species. Journal of Bacteriology 176:6165-6169.

Austin, B., and D. A. Austin. 1987. Bacterial fish pathogens: diseases in farmed and wild fish. Ellis Horwood Limited, England.

Austin, B., and D. Allen-Austin. 1985. A review: bacterial pathogens of fish. Journal of Applied Bacteriology 58:483-506.

Aznar, R., W. Ludwig, R. I. Amann, and K. H. Schleifer. 1994. Sequence determination of rRNA genes of pathogenic Vibrio species and whole-cell identification of Vibrio vulnificus with rRNA-targeted oligonucleotide probes. International Journal of Systematic Bacteriology 44:330-337.

Bhaskar, N., T. M. R. Setty, S. Mondal, M. A. Joseph, C. V. Raju, B. S. Raghunath, and C. S. Anantha. 1998. Prevalence of bacteria of public health significance in the cultured shrimp (Penaeus monodon). Food Microbiology 15:511-519.

Blake, P. A., R. E. Weaver, and D. G. Hollis. 1980. Diseases of humans (other than cholera) caused by vibrios. Annual Review of Microbiology 34:346-367.

Choudhury P., and R. Kumar. 1998. Multidrug- and metal-resistant strains of Klebsiella pneumoniae isolated from Penaeus monodon of the coastal waters of deltaic Sundarban. Canadian Journal of Microbiology 44:186-189.

Costa, A. B., K. Kanai, and K. Yoshikoshi. 1998. Serological characterization of atypical strains of Edwardsiella tarda isolated from sea breams. Fish Pathology 33:265-274.

Dorsch, M., D. Lane, and E. Stackebrandt. 1992. Towards a phylogeny of the genus Vibrio based on 16S rRNA sequences. International Journal of Systematic Bacteriology 42:58-63.

Farrow, J. A. E., P. A. Lawson, H. Hippe, U. Gauglitz, and M. D. Collins. 1995. Phylogenetic evidence that the Gram-negative nonsporulating bacterium Tissierella (Bacteroides) Praeacuta is a member of the Clostridium subphylum of the Gram-positive bacteria and description of Tissierella creatinini sp. nov. International Journal of Systematic Bacteriology 45:436-440.

Felsenstein, J. 1985. Confidence limits on phylogenies: a approch using the bootstrape. Evolution 39:783-791.

Fuerst, J. A., H. G. Gwilliam, M. Lindsay, A. Lichanska, C. Belcher, J. E. Vickers, and P. Hugenholtz. 1997. Isolation and molecular identification of planctomycete bacteria from postlarvae of the giant tiger prawn, Penaeus monodon. Applied and Environmental Microbiology 63: 254-262.

Fuerst, J. A., J. A. Hawkins, A. Holmes, L. I. Sly, C. J. Moore, and E. Stackebrandt. 1993. Porphyrobacter neustonensis gen. nov., sp. nov., an aerobic bacteriochlorophyII-synthesizing budding bacterium from fresh water. International Journal of Systematic Bacteriology 43:125-134.

Hauxhurst, J. D., M. I. Krichevsky, and R. M. Atlas. 1980. Numerical taxonomy of bacteria from the Gulf of Alaska. Journal of General Microbiology 120:131-148.

Hollis, D. G., R. E. Weaver, C. N. Baker, and C. Thornsberry. 1976. Halophilic Vibrio Species isolated from blood cultures. Journal of Clinical Microbiology 3:425-431.

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.

Kita-Tsukamoto, K., H. Oyaizu, K. nanba, and U. Simidu. 1993. Phylogenetic relationships of marine bacteria, mainly member of the family Vibrionaceae, determined on the basis of 16S rRNA sequences. International Journal of Systematic Bactriology 43:8-19.

Krieg, N. R., and J. G. Holt. 1984. Bergey’s manual of systematic bacteriology volume l. p.355-360. Williums & wilkins, USA.

Lavilla-pitogo, C. R., and L. D. de la Pena. 1998. Bacterial diseases in shrimp (Penaeus monodon) culture in the Philippines. Fish Pathology 33:405-411.

Lavilla-Pitogo, C. R., Ma. C. L. Baticados, E. R. Cruz-Lacierda, and L. D. de la Pena. 1990. Occurrence of luminous bacterial disease of Penaeus monodon larvae in the Philippines. Aquaculture 91:1-13.

Lee, K.-K., P.-C. Liu, G.-H. Kou, and S.-N. Chen. 1997. Passive immunization of the tiger prawn, Penaeus monodon, using rabbit antisera to Vibrio harveyi. Letters in Applied Microbiology 25:34-37.

Mullins, T. D., T. B. Britschgi, R. L. Krest, and S. J. Giovannoni. 1995. Genetic comparisons reveal the same unknow bacterial lineages in Atlantic and Pacific bacterioplankton communities. Limnology and Oceanography 40:148-158.

Nix, J., and R. Ingols. 1981. Oxidized manganese from hypolimnetic water as a possible cause of trout mortality in hatcheries. Progressive Fish-Culturist 43:32-36.

O’neill, K. R., S. H. Jones, and D. J. Grimes. 1992. Seasonal incidence of Vibrio vulnificus in the Great Bay estuary of New Hampshire and Maine. Applied and Environmental Microbiology 58:3257-3262.

Pascual, C., P. A. Lawson, J. A. E. Farrow, M. N. Gimenez, and M. D. Collins. 1995. Phylogenetic analysis of the genus Corynebacterium based on 16S rRNA gene sequences. International Journal of Systematic Bacteriology 45:724-728.

Paster, B. J., and F. E. Dewhirst. 1988. Phylogeny of Campylobacters, Wolinellas, Bacteroides gracilis, and Bacteroides ureolyticus by 16S ribosomal ribonucleic acid sequencing. International Journal of Systematic Bacteriology 38:56-62.

Peruski Jr, L. F., and A. H. Peruski. 1997. Analysis of sequence data, p.206-254. In: L. F. Peruski Jr, and A. H. Peruski.(ed.). The internet and the new biology. American Society for Microbiology. Washington, D. C.

Ruangpan, L., P. Na-anan, and S. Direkbusarakom. 1998. Inhibitory effect of Vibrio alginolyticus on the growth of V. hareyi. Fish Pathology 33:293-296.

Ruimy, R., V. Breittmayer, P. Elbaze, B. Lafay, O. Boussemart, M. Gauthier, and R. Christen. 1994. Phylogenetic analysis and assessment of the genera Vibrio, Photobacterium, Aeromonas, and Plesiomonas deduced from small-subunit rRNA sequences. International Journal of Systematic Bactriology 44:416-426.

Saitou, N., and M. Nei. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic tree. Molecular Biological Evolution 4:406-425.

Sako, Y., K. Takai, Y. Ishida, A. Uchida, and Y. Katayama. 1996. Rhodothermus obamensis sp. nov., a modern lineage of extremely thermophilic marine bacteria. International Journal of Systematic Bacteriology 46: 1099-1104.

Sambrook, J. E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning: a laboratory manual (2nded.). Cold Spring Harbor Laboratory Press. New York.

Stackebrandt, E., and M. Goodfellw. 1991. Nucleic acid techniques in bacterial systematics. p.72-73. John Wiley&Sons.

Takami, H., A. Inoue, F. Fuji, and K. Horikoshi. 1997. Microbial flora in the deepest sea mud of the Mariana Trench. FEMS Microbiology Letters 152:279-285.

Thompson, J. D., D. G. Higgins, and T. J. Gibson. 1994. Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Reserch 22:4673-4680.

Vogel, B. F., K. Jorgensen, H. Christensen, J. E. Olsen, and L. Gram. 1997. Differentiation of Shewanella putrefaciens and Shewanella alga on the basis of whole-cell protein profiles, ribotyping, phenotypoic characterization, and 16S rRNA gene sequence analysis. Applied and Environmental Microbiology 63:2189-2199.

Vogt, G. 1997. Hepatopancreatic brush border lysis (HBL)-a new bacterial disease of the shrimp Palaemon elegans. Diseases of Aquatic Organisms 29:151-155.

Weisburg, W. G., S. M. Barns, D. A. Pelletier, and D. J. Lane. 1991. 16S ribosomal DNA amplification for phylogenetic study. Journal Bacteriology 173:697-703.

Willems, A., and M. D. Collins. 1995. Evidence for the placement of the Gram-negative Catonella morbi (Moore and Moore) and Johnsonella ignava (Moore and Moore) within the Clostridium subphylum of the Gram-positive bacteria on the basis of 16S rRNA sequences. International Journal of Systematic Bacteriology 45: 855-857.

王泉傳. 1994. 台灣草蝦養殖池浮游動物相之研究. 國立臺灣大學海洋研究所. 碩士論文.

王憶鎧. 1996. 澳洲球形海綿共生細菌之研究. 國立中山大學海洋資源研究所. 碩士論文.

宋宏紅和蔡福明. 1998. 養殖草蝦肝臟內弧菌組成變化與病害發生的關係. 水產養殖研究報告, 農委會漁業特刊. 第六十二號:159-169.

宋宏紅. 1994. 草蝦抗弧菌的研究. 國立臺灣大學動物學研究所. 博士論文.

李武忠. 1990. 養殖斑節蝦弧菌感染症之研究. 國立臺灣大學漁業科學研究所. 碩士論文.

林義雄, 鍾虎雲和郭光雄. 1989. 台灣水產養殖魚類常見的細菌性病害. 中國水產月刊 433:6-12.

徐玉標, 江漢全和林承民. 1986. 養殖用水之現況調查研究. 水產養殖研究報告彙集(一), 農委會漁業特刊. 第九號:125-137.

徐新亮. 1994. 臺灣草蝦(Penaeus monodon)、淡水長臂大蝦(Macrobrachium rosebergii)、鰻魚(Anguilla japonica)和鱸魚(Micropterus salmoides)之養殖規劃分析. 國立臺灣海洋大學水產養殖研究所. 碩士論文.

張朴性. 1988. 臺灣養殖蝦類之桿狀病毒感染研究. 國立臺灣大學漁業科學研究所. 碩士論文.

張錦宜. 1991. 草蝦(Penaeus monodon)、大正蝦(Penaeus chinensis)養殖池水及蝦肝胰臟內細菌之數值分類研究. 國立臺灣大學漁業科學研究所. 碩士論文.

張簡淑惠. 1995. 眾多指形軟珊瑚共存細菌親緣分析研究. 國立中山大學海洋資源研究所. 碩士論文.

郭玉梅. 1999. 油污分解菌之研究. 國立中山大學海洋資源研究所. 碩士論文.

陳昭德. 1978b. 養殖鰻細菌分佈之研究. 國立臺灣大學動物學研究所. 碩士論文.

陳秋錦. 1990. 草蝦放養密度對養成系統之影響. 國立臺灣海洋大學水產養殖研究所. 碩士論文.

黃文瑛和王毓秀. 1991. 屏東地區蝦池水細菌相之調查研究. 蝦類養殖池環境調查及改善研究, 農委會漁業特刊 第二十八號:285-326.

黃世鈴. 1989. 養殖草蝦細菌感染疾病之研究. 國立臺灣大學漁業科學研究所. 碩士論文.

黃本. 1979. 草蝦池之生態研究. 私立中國文化學院海洋研究所. 碩士論文.

黃銀河. 1977. 虱目魚越冬期間細菌疾病研究-初步報告. JCRR Fisheries Series 29:50-54.

廖一久, 郭光雄, 陳秀男和賴靜誼. 1985. 屏東地區之養殖蝦纇疾病初步調查. 魚病研究專集(七), 農委會漁業特刊 第四號:86-94.

趙維良, 宋宏紅和林秀枝. 1991. 蝦類養殖環境中細菌相之調查. 蝦類養殖池環境調查及改善研究, 農委會漁業特刊 第二十八號:103-122.

趙維良和李海菁. 1998. 草蝦養殖過程中弧菌組成的變化與病害發生的關係. 水產養殖研究報告, 農委會漁業特刊. 第六十二號:171-184.

劉文御. 1995. 台灣養殖蝦池微生物生態學研究. 國立臺灣大學動物學研究所. 博士論文.

蔣先沖. 1991. 四種弧菌對養殖草蝦及斑節蝦之致病性研究. 國立臺灣大學漁業科學研究所. 碩士論文.

鄭文騰. 1989. 臺灣養蝦類病原細菌Vibrio damsela之分離與鑑定. 國立臺灣大學漁業科學研究所. 碩士論文.

謝允廷. 1993. 激發草蝦(Penaeus monodon)血球細胞產生殺菌物質-活性氧族群之分析. 國立臺灣大學漁業科學研究所. 碩士論文.