臺灣博碩士論文加值系統

台灣地處優渥擁有合適的天候條件，多年來累積豐厚的養殖技術，
近年來水產疾病嚴重的影響了養殖產業的發展，以目前產值最高的養
殖魚種石斑魚為例，在育苗的階段即會遭到病毒性神經壞死症（Viral
Nervous Necrosis disease, VNN）的侵害，導致育成率低於一成的慘況，
我們分析了目前對於VNN 防治的研究，提出一個更合適養殖現場使用
的疫苗架構：益生菌複合疫苗，利用能將水體中過量氨氮及亞硝酸進
行降解的Pantoea sp. 及Pseudomonas sp.兩種益生菌中，表現重組神經
壞死病毒外鞘蛋白(Nervous necrosis virus coat protein, NNVCP)，並以
Overlap PCR 技術連接外泌性蛋白Sia10 的DNA 片段，將NNVCP 攜
出益生菌的細胞壁外，實驗證實轉型表現質體之Hero6 和TSR 能夠表
現分子量約為56.2kDa 的融合蛋白Sia10-NNVCP，TSR 能將
Sia10-NNVCP 融合蛋白分泌至細菌培養液中，經免疫螢光染色發現，
此融合蛋白可以崁在細胞的表面。本實驗完成益生菌複合疫苗表現的
基本架構開發，未來我們將進行動物實驗驗證益生菌複合疫苗的保護
效果，並結合冷凍乾燥技術減少益生菌複合疫苗的運送體積、延長菌
種保存時間，期待益生菌複合疫苗能夠克服病毒性神經壞死症常久以
來的問題，並為水產養殖的疾病防治盡一份心力。

Taiwan is a land privileged with optimal climate conditions, and
farmers has cultivated abundant aquafarming techniques over the years.
However,Saquatic diseases have severely affected the growth of the
aquaculture industry in recent years. For example, among grouper fish, the
species with the current highest output value, invasion by the viral nervous
necrosis (VNN) disease begins at the nursery stage. This has led to the
dire state of a breeding rate of less than ten percent. In this research, we
analyzed current studies on VNN disease prevention and suggest a vaccine
more suitable for aquafarm sites, namely, the probiotics combined vaccine.
We used Pantoea sp. and Pseudomonas sp., the two species of probiotics
that can degrade excess ammonia nitrogen and nitrous acid in water, to
rearrange the nervous necrosis virus coat protein (NNVCP). We also used
overlap PCR technology to connect DNA fragments of exocrine protein
Sia10 to NNVCP extracted out of the probiotic cell wall. Studies have
confirmed that transformed expression plasmid Hero6 and TSR can express
the fusion protein Sia10-NNVCP with a molecular weight around 56.2 kDa.
TSR can also secrete Sia10-NNVCP fusion protein into a bacterial culture
medium. Immunofluorescent staining showed that this fusion protein
can be deposited on the surface of the cell. This experiment completed
the basic structural development of the probiotics combined vaccine. In
the future, we will conduct animal testing to verify the protective ability
of the vaccine. We will also incorporate lyophilization to decrease the
delivery volume of the vaccine and extend the preservation time of the
bacteria. We hope the probiotics combined vaccine can overcome the longterm
predicament caused by VNN disease, making our contribution to the
prevention of aquatic diseases.

摘要..............................................................I
Abstract.........................................................II
誌謝.............................................................III
目錄.............................................................IV
壹、前言..........................................................1
一、養殖現況......................................................2
二、氮循環........................................................3
三、益生菌........................................................7
四、病毒性神經壞死症病............................................10
五、魚類的免疫系統................................................13
六、水產類疫苗....................................................15
七、神經壞死病毒疫苗的研發.........................................21
八、研究目的......................................................22
貳、材料與方法....................................................23
一、實驗架構......................................................23
二、實驗材料......................................................24
三、藥劑配方......................................................26
四、實驗儀器設備..................................................27
五、質體和表現DNA片段.............................................28
六、Sia10-NNVCP表現載體備製.......................................28
七、表現Sia10-NNVCP之益生菌製備...................................31
八、蛋白質表現....................................................33
九、免疫螢光染色..................................................34
參、結果.........................................................36
肆、討論.........................................................41
伍、結論.........................................................45
陸、參考文獻.....................................................46
柒、圖...........................................................59
圖1、Overlap PCR實驗架構.........................................59
圖2、EcoRI-Sia10片段.............................................60
圖3、NNVCP-XbaI片段..............................................61
圖4、Sia10-NNVCP片段.............................................62
圖5、EcoRI-Sia10-NNVCP-XbaI片段製備..............................63
圖6、EcoRI-pBBR1MCS-2-XbaI載體...................................64
圖7、EcoRI-Sia10-NNVCP-XbaI片段..................................65
圖8、pBBR1MCS-2-Sia10-RNA2定序結果...............................66
圖9、pBBR1MCS-2-Sia10-RNA2定序結果...............................67
圖10、pBBR1MCS-2-Sia10-NNVCP融合蛋白的結構示意圖..................68
圖11、TSR和Hero6細胞總蛋白之表現分析..............................69
圖12、TSR和Hero6培養基內蛋白分析..................................70
圖13、Pantoea sp.和Hero6中Sia10-NNVCP融合蛋白表現之分析...........71
圖14、Pantoea sp.和Hero6培養基中Sia10-NNVCP融合蛋白表現量分析......72
圖15、Pseudomonas sp. & TSR中Sia10-NNVCP融合蛋白表現量分析........73
圖16、Pseudomonas sp.、TSR培養液中Sia10-NNVCP融合蛋白表現量分析....74
圖17、細菌中Sia10-NNVCP融合蛋白之表現..............................75
圖18、pBBR1MCS-2-Sia10-NNVCP融合蛋白表現示意圖.....................76
捌、表............................................................77
表一、PCR引子總表..................................................77

中文期刊：
田蕴、袁建軍（2007）。南美白對蝦腸道微生物群落的分子分析，微生物學報， 47（4），649-653。
朱家瑩（2009）。抗台灣石斑虹彩病毒蛋粉IgY在石斑魚隻被動保護效果（碩士論文）。國立臺灣海洋大學，基隆市，11-14 pp.。
李國誥（1994）。養殖水產動物細菌性疾病概要，水產動物防疫手冊， 44，41-47。
周信佑（2008）。石斑魚健康種苗之建立與疫苗開發策略，農業生技產業季刊，15，47-50。
和平（2012）。從市售降亞硝酸產品及草蝦腸道中分離及鑑定降亞硝酸之益生菌及其對白蝦生長之影響（碩士論文）。國立高雄海洋科技大學，高雄市，70-75 pp.。
林式修（2003）。魚類免疫與疫苗。水試專訊， 行政院農業委員會，3：14-18。
林思維（2013）。以益生菌作為蝦類白點病口服疫苗開發之研究（碩士論文），國立高雄海洋科技大學，高雄市47-52 pp.。
柯如娜（2010）。食乳酸菌對白蝦之成長、蛋白酶活性和消化道菌量之影響研究（碩士論文）。國立高雄海洋科技大學，高雄市，70-75 pp. 。
胡逸廷（2009）從環境分離和分析鑑定在水產養殖應用上具抑菌能力 之益生菌（碩士論文）。慈濟大學，花蓮縣，57-62 pp. 。
涂堅（1994）。魚類野田病毒之疫情及控制。農政與農情，行政院農業委員會，139：24-28。
翁玉宣（2006）。二價弧菌疫苗應用於養殖石斑魚之研究（碩士論文）。國立臺灣海洋大學，基隆市，42-47 pp. 。
張甘楠（2003）。疫苗發展技術與實驗。動物基因轉殖與疫苗發展技術教學資源中心，41-45。
張照夫（2003）。疫苗發展技術與實驗。動物基因轉殖與疫苗發展技術教學資源中心，台北市，27-34。
張瀚元（2005）。淺談水產養殖，台肥季刊，46(4)，27-33。
陳钰元（2007）。藻酸鈉和鹿角菜膠激活點帶石斑和棕點石斑免疫能力與抵抗Vibrio alginolyticus感染之研究（碩士論文）。國立海洋大學，基隆市，37-42 pp.。
馮忠武與段文龍（2007）。動物生物疫苗。化學工業出版社，北京，4-6。
黃春蘭（2003）。水質學。藝軒圖書出版社，台北市，154-158。
黃琪琰（2001）。水產動物疾病學。水產出版社，基隆市，50-51。
楊惠郎（2003）。疫苗發展技術與實驗。動物基因轉殖與疫苗發展技術教學資源中心，台北市，138-152。
廖一久（2008）。漫談台灣的水產養殖。卓越大師講座。
蓋玉軒（2005）。石斑魚神經壞死症病毒去活化疫苗的製備及效能評估（碩士論文）。國立臺灣大學，台北市，33-38 pp. 。
劉振軒、蔡睦宗、鄭謙仁、張本恒、王汎熒、洪信雄、徐榮彬（2002）。動物傳染病附冊。行政院業委員會動植物防疫檢疫局，278-281。
蔡喬欣（2011）。養殖密度、換水率及添加光合菌(Rhodobium sp.)和藻水對於羅氏沼蝦(Macrobrachium rosenbergii)的成長、活存及水質因子之影響（碩士論文）。國立東華大學，花蓮縣，63-68 pp. 。
賴威延、許仁弘、陳添進、吳金洌（2011）。台灣石斑魚研發能量與優勢。農業生技產業季刊，26，51-57。
謝懷貞（2008）。春夏季貧營養黑潮海域浮游細菌群聚組成之變化（碩士論文）。國立臺灣師範大學，台北市，37-42 pp. 。

英文期刊：
Amal, M. N. A., Saad, M. Z., Zahrah, A. S., & Zulkafli, A. R. (2015). Water quality influences the presence of Streptococcus agalactiae in cage cultured red hybrid tilapia, Oreochromis niloticus× Oreochromis mossambicus. Aquaculture Research, 46(2), 313-323.
Arasteh, N., Aminirissehei, A. H., Yousif, A., Albright, L. J., & Durance, T. D. (2004). Passive immunization of rainbow trout (Oncorhynchus mykiss) with chicken egg yolk immunoglobulins (IgY). Aquaculture, 231(1-4), 23-36.
Arimoto, M., Mushiake, K., Mizuta, Y., Nakai, T., Muroga, K., & Furusawa, I. (1992). Detection of Striped Jack Nervous Necrosis Virus (Sjnnv) by Enzyme-Linked-Immunosorbent-Assay (Elisa). Fish Pathology, 27(4), 191-195.
Asis, C. A., Jr. & Adachi, K. (2004). Isolation of endophytic diazotroph Pantoea agglomerans and nondiazotroph Enterobacter asburiae from sweetpotato stem in Japan. Letters in Applied Microbiology, 38(1), 19-23.
Blazer, V. (1991). Piscine Macrophage Function and Nutritional Influences: A Review. Journal of Aquatic Animal Health, 3(2), 77-86.
Bloch, B., Gravningen, K., & Larsen, J. L. (1991). Encephalomyelitis among turbot associated with a picornavirus-like agent. Diseases of Aquatic Organisms, 10(1), 65-70.
Brady, C. L., Venter, S. N., Cleenwerck, I., Engelbeen, K., Vancanneyt, M., Swings, J., & Coutinho, T. A. (2009). Pantoea vagans sp. nov., Pantoea eucalypti sp. nov., Pantoea deleyi sp. nov. and Pantoea anthophila sp. nov. International Journal of Systematic and Evolutionary Microbiology, 59(9), 2339-2345.
Brock, J. A. & Bullis, R. (2001). Disease prevention and control for gametes and embryos of fish and marine shrimp. Aquaculture, 197(1-4), 137-159.
Carbajal-Gonzalez, M. T., Fregeneda-Grandes, J. M., Suarez-Ramos, S., Rodriguez Cadenas, F., & Aller-Gancedo, J. M. (2011). Bacterial skin flora variation and in vitro inhibitory activity against Saprolegnia parasitica in brown and rainbow trout. Diseases of Aquatic Organisms, 96(2), 125-135.
Chi, S. C., Lo, B. J. & Lin, S. C. (2001). Characterization of grouper nervous necrosis virus (GNNV). Journal of Fish Diseases, 24(1), 3-13.
Costa, E., Teixidó, N., Usall, J., Atarés, E., & Viñas, I. . (2002). The effect of nitrogen and carbon sources on growth of the biocontrol agent Pantoea agglomerans strain CPA‐2. Letters in applied microbiology, 35(2), 117-120.
El-Shafei, K., Sharaf, M., Ibrahim, A., Sadek, I., & El-Sayed, S. (2013). Evaluation of Lactobacillus plantarum as a probiotic in aquaculture: Emphasis on growth performance and innate immunity. Journal of Applied Sciences Research, 9(1), 572-582.
Ellis, A. E. (1988). Fish vaccination (No. 4). London: Academic press. 255 pp..
Fenner, B. J., Thiagarajan, R., Chua, H. K., & Kwang, J. (2006). Betanodavirus B2 is an RNA interference antagonist that facilitates intracellular viral RNA accumulation. Journal of virology, 80(1), 85-94.
Fouzi, M., Shariff, M., Omar, A. R., Yusoff, F.M., & Tan, S. W. (2010). TaqMan real‐time PCR assay for relative quantification of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia. Journal of fish diseases, 33(12), 931-938.
Fryer, J. L., Rohovec, J. S., Tebbit, G. L., McMichael, J. S., & Pilcher, K. S. (1976). Vaccination for control of infectious diseases in Pacific salmon. Fish Pathology, 10(2), 155-164.
Gatesoupe, F. J. (1999). The use of probiotics in aquaculture. Aquaculture, 180(1), 147-165.
Glazebrook, J. S., Heasman, M. P., & De Beer, S. W. (1990). Picorna-like viral particles associated with mass mortalities in larval barramundi, Lates calcarifer Bloch. Journal of Fish Diseases, 13(3), 245-249.
Gram, L., Melchiorsen, J., Spanggaard, B., Huber, I., & Nielsen, T. F. (1999). Inhibition of Vibrio anguillarum by Pseudomonas fluorescens AH2, a possible probiotic treatment of fish. Applied and Environmental Microbiology, 65(3), 969-973.
Guo, H., Xian, J. A., Li, B., Ye, C. X., Wang, A. L., Miao, Y. T., & Liao, S. A. (2013). Gene expression of apoptosis-related genes, stress protein and antioxidant enzymes in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 157(4), 366-371.
Guo, Y. X., Chan, S. W., & Kwang, J. (2004). Membrane association of greasy grouper nervous necrosis virus protein A and characterization of its mitochondrial localization targeting signal. Journal of virology, 78(12), 6498-6508.
Gutierrez, M. A., Miyazaki, T., Hatta, H., & Kim, M. (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(2), 113-122.
Hansen, G. H. & Olafsen, J. A. (1999). Bacterial interactions in early life stages of marine cold water fish. Microbial ecology, 38(1), 1-26.
Hoque, F. (2014). Biocontrol of ß-haemolytic Aeromonas hydrophila infection in Labeo Rohita using antagonistic bacterium Pseudomonas aeruginosa FARP72. International Journal of Pharmaceutical Sciences & Research, 5(2), 409-501.
Hu, X. J., Li, Z. J., Cao, Y. C., Zhang, J., Gong, Y. X., & Yang, Y. F. (2010). Isolation and identification of a phosphate-solubilizing bacterium Pantoea stewartii subsp. stewartii g6, and effects of temperature, salinity, and pH on its growth under indoor culture conditions. Aquaculture International, 18(6), 1079-1091.
Isnansetyo, A. & Kamei, Y. (2009). Bioactive substances produced by marine isolates of Pseudomonas. Journal of industrial microbiology & biotechnology, 36(10), 1239-1248.
Jenkins, P. G., Wrathmell, A. B., Harris, J. E., & Pulsford, A. L. (1994). The effects of different adjuvants on intestinal antigen absorption and subsequent immune responses of the tilapian Oreochromis mossambicus. Fish Shellfish Immunology, 4(3), 167-177.
Körner, H. & Zumft, W. G. (1989). Expression of denitrification enzymes in response to the dissolved oxygen level and respiratory substrate in continuous culture of Pseudomonas stutzeri. Appl Environ Microbiol, 55(7), 1670-1676.
Kai, Y. H., & Chi, S. C. (2008). Efficacies of inactivated vaccines against betanodavirus in grouper larvae (Epinephelus coioides) by bath immunization. Vaccine, 26(11), 1450-1457.
Kovach, M. E., Elzer, P. H., Hill, D. S., Robertson, G. T., Farris, M. A., Roop, R. M., & Peterson, K. M. (1995). Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene, 166(1), 175-176.
Lilly, D. M. & Stillwell, R. H. (1965). Probiotics: growth-promoting factors produced by microorganisms. Science, 147(3659), 747-748.
Lin, C. C., Lin, J. H. Y., Chen, M. S., & Yang, H. L. (2007). An oral nervous necrosis virus vaccine that induces protective immunity in larvae of grouper (Epinephelus coioides). Aquaculture, 268(1-4), 265-273.
Liu, W. T., Hsu, C. H., Chang, C. Y., Chen, H. H., & Lin, C. S. (2006). Immune response against grouper nervous necrosis virus by vaccination of virus-like particles. Vaccine, 24(37-39), 6282-6287.
Loiret, F. G., Ortega, E., Kleiner, D., Ortega-Rodes, P., Rodes, R., & Dong, Z. (2004). A putative new endophytic nitrogen-fixing bacterium Pantoea sp. from sugarcane. Journal of applied microbiology, 97(3), 504-511.
Lorenzen, N., Olesen, N. J., Jorgensen, P. E., Etzerodt, M., Holtet, T. L., & Thogersen, H. C. (1993). Molecular cloning and expression in Escherichia coli of the glycoprotein gene of VHS virus, and immunization of rainbow trout with the recombinant protein. Journal of General Virology, 74 (4), 623-630.
Lu, M. W., Liu, W. T., & Lin, C. S. (2003). Infection competition against grouper nervous necrosis virus by virus-like particles produced in Escherichia coli. Journal of General Virology, 84(6), 1577-1582.
Manning, D. S. & Leong, J. C. (1990). Expression in Escherichia coli of the large genomic segment of infectious pancreatic necrosis virus. Virology, 179(1), 16-25.
Mori, K. I., Nakai, T., Muroga, K., Arimoto, M., Mushiake, K., & Furusawa, I. (1992). Properties of a new virus belonging to Nodaviridae found in larval striped jack (Pseudocaranx dentex) with nervous necrosis. Virology, 187(1), 368-371.
Munday, B. L. & Nakai, T. (1997). Nodaviruses as pathogens in larval and juvenile marine finfish. World Journal of Microbiology and Biotechnology, 13(4), 375-381.
Mushiake, K., Nishizawa, T., Nakai, T., Furusawa, I., & Muroga, K. (1994). Control of VNN in striped jack: selection of spawners based on the detection of SJNNV gene by polymerase chain reaction (PCR). Fish Pathology, 29(3), 177-182.
Nakajima, K., Maeno, Y., Kurita, J., & Inui, Y. (1997). Vaccination against red sea bream iridoviral disease in red sea bream. Fish Pathology, 32(4), 205-209.
Nakanishi, T. & Ototake, M. (1997). Antigen uptake and immune responses after immersion vaccination. Developments in Biological Standardization, 90, 59-68.
Nishizawa, T., Mori, K., Furuhashi, M., Nakai, T., Furusawa, I., & Muroga, K. (1995). Comparison of the coat protein genes of five fish nodaviruses, the causative agents of viral nervous necrosis in marine fish. Journal of General Virology, 76 (7), 1563-1569.
Popp, A., Cleenwerck, I., Iversen, C., De Vos, P., & Stephan, R. (2010). Pantoea gaviniae sp. nov. and Pantoea calida sp. nov., isolated from infant formula and an infant formula production environment. International Journal of Systematic and Evolutionary Microbiology, 60(12), 2786-2792.
Rengpipat, S., Phianphak, W., Piyatiratitivorakul, S., & Menasveta, P. (1998). Effects of a probiotic bacterium on black tiger shrimp Penaeus monodon survival and growth. Aquaculture, 167(3), 301-313.
Roitt, I. M. (1984). The acquired immune response. Essential Immunology, 134-153.
Rungrassamee, W., Maibunkaew, S., Karoonuthaisiri, N., & Jiravanichpaisal, P. (2013). Application of bacterial lipopolysaccharide to improve survival of the black tiger shrimp after Vibrio harveyi exposure. Developmental & Comparative Immunology, 41(2), 257-262.
Salvesen, I. & Vadstein, O. (1995). Surface disinfection of eggs from marine fish: evaluation of four chemicals. Aquaculture International, 3(3), 155-171.
Scapigliati, G., Romano, N., Buonocore, F., Picchietti, S., Baldassini, M. R., Prugnoli, D.＆Abelli, L. (2002). The immune system of sea bass, Dicentrarchus labrax, reared in aquaculture. Developmental & Comparative Immunology, 26(2), 151-160.
Secombes, C. J. & Ellis, A.E. (2012) The immunology of teleosts. In: Fish pathology. Wiley-Blackwell, Oxford, UK, pp 144–166.
Skalli, A., Castillo, M., Andree, K. B., Tort, L., Furones, D., & Gisbert, E. (2013). The LPS derived from the cell walls of the Gram-negative bacteria Pantoea agglomerans stimulates growth and immune status of rainbow trout (Oncorhynchus mykiss) juveniles. Aquaculture, 416, 272-279.
Skliris, G. P. & Richards, R. H. (1999). Induction of nodavirus disease in seabass, Dicentrarchus labrax, using different infection models. Virus Research, 63(1-2), 85-93.
Sommerset, I. & Nerland, A. H. (2004). Complete sequence of RNA1 and subgenomic RNA3 of Atlantic halibut nodavirus (AHNV). Diseases of Aquatic Organisms, 58(2-3), 117-125.
Sun, B. G., Dang, W., Sun, L., & Hu, Y. H. (2014). Vibrio harveyi Hsp70: Immunogenicity and application in the development of an experimental vaccine against V. harveyi and Streptococcus iniae. Aquaculture, 418, 144-147.
Svobodova, Z., Pravda, D. & Palackova, J. (1991): Unified methods of haematological examination of fish (in Czech). Methods No. 22, Research Institute of Fish Culture and Hydrobiology, Vodnany, 31 pp.
Tacon, A. G. J., Cody, J. J., Conquest, L. D., Divakaran, S., Forster, I. P., & Decamp, O. E. (2002). Effect of culture system on the nutrition andgrowth performance of Pacific white shrimp Litopenaeus vannamei (Boone) fed different diets. Aquaculture Nutrition, 8(2), 121-137.
Tanaka, S., Mori, K., Arimoto, M., Iwamoto, T., & Nakai, T. (2001). Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis. Journal of Fish Diseases, 24(1), 15-22.
Tang, L., Lin, C. S., Krishna, N. K., Yeager, M., Schneemann, A. & Johnson, J. E. (2002). Virus-like particles of a fish nodavirus display a capsid subunit domain organization different from that of insect nodaviruses. Journal of Virology, 76(12), 6370-6375.
Tomasso, J. R. (1994). Toxicity of nitrogenous wastes to aquaculture animals. Reviews in Fisheries Science, 2(4), 291-314.
Tomasso, J. R. (2012). Environmental nitrite and aquaculture: a perspective. Aquaculture International, 20(6), 1107-1116.
Vaseeharan, B. A. R. P. & Ramasamy, P. (2003). Control of pathogenic Vibrio spp. by Bacillus subtilis BT23, a possible probiotic treatment for black tiger shrimp Penaeus monodon. Letters in applied microbiology, 36(2), 83-87.
Jorgensen, P. E. V. (1976). Partial resistance of rainbow trout (Salmo gairdneri) to viral haemorrhagic septicaemia (VHS) following exposure to non-virulent Egtved virus. Nordisk veterinaermedicin, 28(11), 570-571.
Yoshikoshi, K. & Inoue, K. (1990). Viral nervous necrosis in hatchery-reared larvae and juveniles of Japanese parrotfish, Oplegnathus fasciatus (Temminck & Schlegel). Journal of Fish Diseases, 13(1), 69-77.
Yusoff, F. M., Banerjee, S., Khatoon, H., & Shariff, M. (2001). Biological approaches in management of nitrogenous compounds in aquaculture systems. Received: 19 January, 2010. Accepted: 5 Dynamic Biochemistry, Process Biotechnology and Molecular Biology, 5, 21-31.
Zhen, J., Zhang, X., Hu, C., & Li, W. (2013). Effect of Pseudomonas stutzeri F1M on water quality in grass carp culture. Fishery Modernization, 3, 5-9.
Zhou, M. H., Ye, H. R., & Zhao, X. W. (2014). Isolation and characterization of a novel heterotrophic nitrifying and aerobic denitrifying bacterium Pseudomonas stutzeri KTB for bioremediation of wastewater. Biotechnology and Bioprocess Engineering, 19(2), 231-238.
Zhu, D., Wang, G., Qiao, H., & Cai, J. (2008). Fermentative hydrogen production by the new marine Pantoea agglomerans isolated from the mangrove sludge. International Journal of Hydrogen Energy, 33(21), 6116-6123.
Ziaei-Nejad, S., Rezaei, M. H., Takami, G. A., Lovett, D. L., Mirvaghefi, A. R., & Shakouri, M. (2006). The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian white shrimp Fenneropenaeus indicus. Aquaculture, 252(2), 516-524.