(3.238.174.50) 您好!臺灣時間:2021/04/18 02:37
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:陳偉元
論文名稱:分離與鑑定可分解Citrobacter freundii 的噬菌體
論文名稱(外文):Isolation and identification of a bacteriophage that can lyse Citrobacter freundii
指導教授:張文興
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:生物農業科技學系研究所
學門:農業科學學門
學類:農業技術學類
論文種類:學術論文
畢業學年度:104
語文別:中文
中文關鍵詞:吳郭魚弗氏檸檬酸桿菌噬菌體噬菌體療法次世代定序
外文關鍵詞:TilapiaCitrobacter freundiiWY-1 bacteriophagePhage therapyNext generation sequencing
相關次數:
  • 被引用被引用:0
  • 點閱點閱:246
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:16
  • 收藏至我的研究室書目清單書目收藏:0
因為氣候變遷、環境污染和過度捕撈,造成全球撈捕產量成長停滯,水產養殖則因為技術及設備的提升而快速發展。而因為地球人口暴漲,對魚類蛋白質的需求大幅增加,使水產養殖供給人類需求的比重逐年增加。吳郭魚因其環境需求低、成長快速及雜食性,使之成為廣泛養殖的魚種,但也因為高密度的水產養殖,爆發出許多疫情。奠基於本實驗室分離出一株會造成吳郭魚死亡的致病菌,弗氏檸檬酸桿菌 (Citrobacter freundii),我們已經分離並鑑定一株能專一性感染弗氏檸檬酸桿菌的噬菌體 (Citrobacter freundii phage WY-1, WY-1),冀望研究噬菌體療法 (phage therapy) 的可行性,使之能夠替代抗生素,控制本菌的感染。在本實驗中,以次世代定序 (Next generation sequencing, NGS) 的方式,得到 WY-1 的基因體序列,大小約 59 kb,並確認噬菌體 WY-1 為線性雙股 DNA 病毒。在一步生長曲線實驗中,WY-1 的潛伏時間為 25 分鐘,裂解量 (burst size) 為 105 PFU/cell。對溫度耐性,WY-1 的活性在 80℃ 時無法測得。對酸鹼耐性,為 pH 4.0-10.0。鹽濃度幾乎不影響噬菌體的活性,越低溫越利於保存。當利用 WY-1 體外感染弗氏檸檬酸桿菌時,MOI 越高,裂解力越高;而 WY-1 不感染E.coli XL-1-Blue。在攻毒試驗中,弗氏檸檬酸桿菌感染造成吳郭魚極高的致死率,弗氏檸檬酸桿菌的半致死劑量 (lethal dose 50%, LD50) 為 84 CFU,噬菌體 WY-1半保護劑量 (protective dose 50% ,PD50) 為 2.473 ×108 PFU。這些發現能為吳郭魚的噬菌體療法應用,提供有用的資訊。
Because of climate change, environmental pollution, and overfishing, global marine fish capture production growth has stagnated. Owing to its low environment requirements, fast growth, and being omnivorous, tilapia is currently widely cultured to meet the growing demand of fish meat. However, high density aquaculture has caused many outbreaks of tilapia disease. A tilapia pathogen, Citrobacter freundii, has been isolated and confirmed to cause the death of tilapia. In this thesis, a bacteriophage (Citrobacter freundii phage WY-1, WY-1) is isolated, analyzed by next-generation sequencing, and characterized to have specificity toward Citrobacter freundii. Feasibility study of phage therapy hoping to replace antibiotics is also tested in this report. Data from Next-generation sequencing analysis indicate WY-1’s genome size is 59 kb. By nuclease digestion assay, the WY-1 is confirmed to be a linear double stranded DNA virus. By the one step growth curve experiment, WY-1 exhibits a latent period of 25 min and burst size of 105 PFU/cell. Moreover, thermal and pH stability tests of WY-1 phage indicate that phage will die at 80℃ and survive well at the pH ranged from 4.0 to 10.0. Changes of salt concentration seem not to affect the viability of WY-1 phage. In vitro infection assays of Citrobacter freundii by WY-1 indicate that higher MOI shows higher lysis ability. Furthermore, WY-1 is found not to infect E.coli XL-1-Blue. By pathogen challenge assays, tilapia infected by Citrobacter freundii can cause extremely high mortality rate. The median lethal dose of Citrobacter freundii for killing tilapia is calculated to be 84 CFU/ml. The WY-1 semi protecting dose is calculated to be 2.473 ×108 PFU/ml. These findings can help to apply WY-1 as a useful tool for preventing tilapia from Citrobacter freundii infection.
摘要…………………………………………………………………………………………………………………………………………I
Abstrate……………………………………………………………………………………………………………………………II
誌謝………………………………………………………………………………………………………………………………………IV
目錄………………………………………………………………………………………………………………………………………V
圖片目錄………………………………………………………………………………………………………………………VIII
表格目錄……………………………………………………………………………………………………………………………IX
1.前言……………………………………………………………………………………………………………………………………1
1.1.全球水產養殖現況………………………………………………………………………………………………1
1.2.吳郭魚 (Tilapia) 之簡介……………………………………………………………………………3
1.2.1.吳郭魚的種類與分布………………………………………………………………………………3
1.2.2.吳郭魚的養殖優勢……………………………………………………………………………………4
1.2.3.吳郭魚的疾病………………………………………………………………………………………………5
1.3.噬菌體簡介………………………………………………………………………………………………………………7
1.3.1.噬菌體的發現………………………………………………………………………………………………7
1.3.2.噬菌體和細菌的關係………………………………………………………………………………8
1.4.噬菌體療法 (phage therapy)…………………………………………………………………9
1.4.1.噬菌體療法簡介…………………………………………………………………………………………9
1.4.2.噬菌體療法和抗生素療法的比較……………………………………………………10
1.4.3.噬菌體療法應用在魚類………………………………………………………………………10
1.4.4.噬菌體療法的其他應用………………………………………………………………………11
1.5.次世代定序 (Next Generation Sequencing, NGS)………………13
1.6.實驗目的與目標………………………………………………………………………………………………14
2.實驗步驟與方法………………………………………………………………………………………………………15
2.1.菌株來源與生長曲線……………………………………………………………………………………15
2.1.1.菌株來源…………………………………………………………………………………………………15
2.1.2.細菌之培養與保存………………………………………………………………………………15
2.1.3.分光光度計 (spectrophotometer) 測定細菌生長曲線…16
2.1.4.稀釋平板法 (dilution plate method) 測定細菌生長曲
線………………………………………………………………………………………………………………16
2.2.噬菌體增量、分離與保存…………………………………………………………………………17
2.2.1.噬菌體增量 (phage enrichment)…………………………………………17
2.2.2.噬菌體分離……………………………………………………………………………………………18
2.2.3.噬菌體大量純化保存…………………………………………………………………………19
2.3.分離噬菌體基因組 DNA (genomic DNA)…………………………………………20
2.3.1 石碳酸-氯仿萃取法(phenol-chloroform extraction)萃
取噬菌體基因組 DNA ………………………………………………………………………20
2.3.2.核酸膠體電泳………………………………………………… 21
2.3.3.分析噬菌體基因組核酸是否為 DNA ……………………………………………22
2.3.4.噬菌體基因組 DNA 限制性內切酶分析(Restriction endonuclease analysis)……………………………………………………………………………………23
2.3.5.噬菌體 DNA 線狀或環狀分析…………………………………………………………24
2.4.噬菌體基因組片段的選殖及核酸序列分析…………………………………………25
2.4.1.膠體純化…………………………………………………………………………………………………25
2.4.2.噬菌體基因片段與載體之接合作用(ligation)…………………26
2.4.3.重組質體的轉型作用 (transformation)……………………………27
2.4.4.重組質體之挑選確認…………………………………………………………………………28
2.4.5.重組質體之酵素切割確認………………………………………………………………29
2.4.6.質體 DNA 定序……………………………………………………………………………………30
2.5.噬菌體基因組次世代定序 (NGS) 分析………………………………………………30
2.6.噬菌體生理特性分析……………………………………………………………………………………32
2.6.1.一步生長曲線 (one step growth cruve)…………………………32
2.6.2.噬菌體對溫度的耐受性………………………………………………………………………33
2.6.3.噬菌體對 pH 值的耐受性………………………………………………………………33
2.6.4.噬菌體對鹽濃度的耐受性…………………………………………………………………34
2.6.5.噬菌體保存條件……………………………………………………………………………………35
2.6.6.細菌裂解能力(bacteriolytic activity) 分析(液態培養
法)………………………………………………………………………………………………………………35
2.6.7.細菌裂解能力(bacteriolytic activity) 分析(固態培養
法)………………………………………………………………………………………………………………36
2.6.8.噬菌體宿主範圍 (host range)…………………………………………………37
2.7.動物實驗測試……………………………………………………………………………………………………37
2.7.1.全菌攻毒試驗…………………………………………………………………………………………38
2.7.2.弗氏檸檬酸桿菌半致死劑量 (Lethal Dose 50%, LD50)
………………………………………………………………………………………………………………………38
2.7.3.噬菌體療法半保護力劑量試驗 (Protective Dose
50%,PD50)………………………………………………………………………………………………40
3.結果………………………………………………………………………………………………………………………………42
3.1.弗氏檸檬酸桿菌 (Citrobacter freundii) 生長曲線……………42
3.1.1.弗氏檸檬酸桿菌生長曲線 (濁度測定法)…………………………………42
3.1.2.弗氏檸檬酸桿菌生長曲線 (稀釋平板法)…………………………………43
3.2.噬菌體分離…………………………………………………………………………………………………………44
3.3.噬菌體物種鑑定………………………………………………………………………………………………46
3.3.1.噬菌體核酸形式電泳分析…………………………………………………………………46
3.3.2.噬菌體核酸限制酶分析………………………………………………………………………47
3.3.3.噬菌體 DNA 線狀或環狀分析…………………………………………………………48
3.3.4.噬菌體基因組次世代定序 (NGS) 分析結果……………………………50
3.3.5.生物資訊分析…………………………………………………………………………………………53
3.4.噬菌體的特性分析…………………………………………………………………………………………53
3.4.1.噬菌體的一步生長曲線 (one step growth curve)……53
3.4.2.噬菌體對溫度的耐受性……………………………………………………………………55
3.4.3.噬菌體對 pH 值的耐受性………………………………………………………………56
3.4.4.噬菌體對鹽濃的耐受性……………………………………………………………………57
3.4.5.噬菌體保存條件…………………………………………………………………………………57
3.4.6.噬菌體 WY-1對弗氏檸檬酸桿菌的裂解能力 (液態培養法)
……………………………………………………………………………………………………………………59
3.4.7.噬菌體 WY-1對弗氏檸檬酸桿菌的裂解能力 (固態培養法)
……………………………………………………………………………………………………………………60
3.4.8.噬菌體宿主範圍…………………………………………………………………………………61
3.5.吳郭魚攻毒試驗……………………………………………………………………………………………63
3.5.1.全菌攻毒試驗………………………………………………………………………………………63
3.5.2.弗氏檸檬酸桿菌半致死劑量 (Lethal Dose 50%, LD50)
……………………………………………………………………………………………………………………64
3.5.3.噬菌體 WY-1 半保護劑量試驗 (Protective Dose 50%
,PD50)……………………………………………………………………………………………………66
4.討論……………………………………………………………………………………………………………………………70
4.1.從環境中分離噬菌體…………………………………………………………………………………70
4.2.噬菌體的生理特性對於應用的價值……………………………………………………71
4.3.次世代定序的價值………………………………………………………………………………………72
4.4.吳郭魚噬菌體攻毒試驗分析……………………………………………………………………73
4.4.1.環境因子………………………………………………………………………………………………73
4.4.2.噬菌體攻毒試驗與魚體反應………………………………………………………74
4.4.3 噬菌體療法的投藥…………………………………………………………………………75
4.5.噬菌體療法的未來……………………………………………………………………………………75
4.6.結論…………………………………………………………………………………………………………………76
5.參考文獻………………………………………………………………………………………………………………78
附錄一、藥品…………………………………………………………………………………………………………84
附錄二、DNA marker ………………………………………………………………………………………85
附錄三、噬菌體 WY-1 全基因組定序結果………………………………………………86
附錄四、Predict BlastN analysis …………………………………………………101
附錄五、Predict BlastX analysis……………………………………………………104
1.Adhya, S., & Merril, C. (2006). The road to phage therapy. Nature, 443(7113),754-755.
2.Amin, N. E., Abdallah, I. S., Faisal, M., Easa, M. E. S., Alaway, T., & Alyan, S. A. (1988).Columnaris infection among cultured Nile tilapia Oreochromis niloticus. Antonie van Leeuwenhoek, 54(6), 509-520.
3.Atterbury, R. J., Connerton, P. L., Dodd, C. E., Rees, C. E., & Connerton, I. F. (2003). Application of host-specific bacteriophages to the surface of chicken skin leads to a reduction in recovery of Campylobacter jejuni. Applied and environmental microbiology, 69(10), 6302-6306.
4.Balarin, J. D., & Haller, R. D. (1982). The intensive culture of tilapia in tanks, raceways and cages. Recent advances in aquaculture, 266-355.
5.Barrow, P. A., & Soothill, J. S. (1997). Bacteriophage therapy and prophylaxis: rediscovery and renewed assessment of potential. Trends in microbiology, 5(7), 268-271.
6.Bergh, Ø.,BØrsheim, K. Y., Bratbak, G., & Heldal, M. (1989). High abundance of viruses found in aquatic environments. Nature, 340(6233), 467-468.
7.Brüssow, H., & Hendrix, R. W. (2002). Phage genomics: small is beautiful.Cell, 108(1), 13-16.
8.Bruttin, A., & Brüssow, H. (2005). Human volunteers receiving Escherichia coli phage T4 orally: a safety test of phage therapy. Antimicrobial agents and chemotherapy, 49(7), 2874-2878.
9.Bruynoghe, R. A. J. M., & Maisin, J. (1921). Essais de thérapeutique au moyen du bacteriophage. CR Soc. Biol, 85, 1120-1121.
10.Bunch, E.C., & Bejerano, I. (1997). The effect of environmental factors on the susceptibility of hybrid tilapia, Oreohromis niloticus×Oreohromis aureus, to streptococcosis. Israeli Journal of aquaculture/Bamidgeh, 49, 67-76.
11.Canagaratnam, P. (1966). Growth of Tilapia mossambica Peters in different salinities. Bulletin of the Fisheries Research Station of Ceylon, 19, 1–2.
12.Chanishvili, N. (2012). Advances in Virus Research. Elsevier, 83, 4- 10.
13.Chaudhry, W. N., Haq, I. U., Andleeb, S., & Qadri, I. (2014). Characterization of a virulent bacteriophage LK1 specific for Citrobacter freundii isolated from sewage water. Journal of basic microbiology, 54(6), 531-541.
14.Chernomordik, A. B. (1989). Bacteriophages and their therapeutic-prophylactic use. Meditsinskaiasestra, 48(6), 44-47.
15.Chervinski, J. (1982). Environmental physiology of tilapias. In: The biology and culture of tilapias. International Center for Living Aquatic Resources Management Manila, Philippines, 7, 119-128.
16.Everitt,S., & Leung, C.-F.A. (1999). Fish disease in Hong Kong. Paper presented at the Fourth Symposium on Diseases in Asian Aquaculture: Aquatic Animal Health for Sustainability.November, Cebu City, Philippines. 22-26.
17.Farrah, S. R. (1987). Ecology of phage in freshwater environments. Phage Ecology, 125-135.
18.F. A. O. Fisheries and Aquaculture Department. (2014). The state of world fisheries and aquaculture. Food and Agriculture Organization of the United Nations, Rome.
19.F. A. O. Yearbook. (2012). Fishery and Aquaculture Statistics. Food and Agriculture Organization of the United Nations. Rome, 219.
20.Filée, J., Tétart, F., Suttle, C. A., & Krisch, H. M. (2005). Marine T4-type bacteriophages, a ubiquitous component of the dark matter of the biosphere. Proceedings of the National Academy of Sciences of the United States of America, 102(35), 12471-12476.
21.Garrity, G.M., (2005). The Proteobacteria. Bergeys’s Manaul of Systematic Bacteriology, Springer, New York.
22.Getachew, T. (1989). Stomach pH, feeding rhythm and ingestion rate in Oreochromis niloticus L.(Pisces: Cichlidae) in Lake Awasa, Ethiopia.Hydrobiologia, 174(1), 43-48.
23.Grada, A., & Weinbrecht, K. (2013). Next-generation sequencing: methodology and application. Journal of Investigative Dermatology, 133(8), e11.
24.Harshey, R. M., & Bukhari, A. I. (1983). Infecting bacteriophage Mu DNA forms a circular DNA-protein complex. Journal of molecular biology, 167(2), 427-441.
25.Helton, R. R., Liu, L., & Wommack, K. E. (2006). Assessment of factors influencing direct enumeration of viruses within estuarine sediments. Applied and environmental microbiology, 72(7), 4767-4774.
26.Henson, J., Tischler, G., & Ning, Z. (2012). Next-generation sequencing and large genome assemblies. Pharmacogenomics, 13(8), 901-915.
27.Huang, S. L., Chen, W. C., Shei, M. C., Liao, I. C., & Chen, S. N. (1999). Studies on epizootiology and pathogenicity of Staphylococcus epidermidis in tilapia (Oreochromis spp.) cultured in Taiwan. Zoological Studies-Taipei, 38, 178-188.
28.Huang, S. L., Liao, I. C., & Chen, S. N. (2000). Induction of apoptosis in tilapia, Oreochromis aureus Steindachner, and in TO‐2 cells by Staphylococcus epidermidis. Journal of Fish Diseases, 23(6), 363-368.
29.Jobling, M., & Hjelmeland, K. (1992). Ernaering og metabolisme. Fiskens Fysiologi. John Grieg Forlag, Bergen, Norway, 234-276.
30.Leverentz, B., Conway, W. S., Alavidze,Z., Janisiewicz, W. J., Fuchs, Y., Camp, M. J., Chighladze, E., & Sulakvelidze, A. (2001). Examination of bacteriophage as a biocontrol method for Salmonella on fresh-cut fruit: a model study. Journal of Food Protection®, 64(8), 1116-1121.
31.Leverentz, B., Conway, W. S., Camp, M. J., Janisiewicz, W. J., Abuladze, T., Yang, M., Saftner, R. & Sulakvelidze, A. (2003). Biocontrol of Listeria monocytogenes on fresh-cut produce by treatment with lytic bacteriophages and a bacteriocin. Applied and Environmental Microbiology, 69(8), 4519-4526.
32.Libey, George S., & Timmons, M. B. (1998). Proceedings of the Second International Conference on Recirculating Aquaculture. Blacksburg, VA :Cooperative Extension/Sea Grant, Virginia Tech.
33.Madsen, L., Bertelsen, S. K., Dalsgaard, I., & Middelboe, M. (2013). Dispersal and survival of Flavobacterium psychrophilum phages in vivo in rainbow trout and in vitro under laboratory conditions: implications for their use in phage therapy. Applied and environmental microbiology, 79(16), 4853-4861.
34.Mardis, E. R. (2008). Next-generation DNA sequencing methods. Annu. Rev. Genomics Hum. Genet., 9, 387-402.
35.Matsuzaki, S., Rashel, M., Uchiyama, J., Sakurai, S., Ujihara, T., Kuroda, M., Ikeuchi, M., Tani, T., Fujieda, M., Wakiguchi, H., & Imai, S. (2005). Bacteriophage therapy: a revitalized therapy against bacterial infectious diseases. Journal of infection and chemotherapy, 11(5), 211-219.
36.Matsuzaki, S., Yasuda, M., Nishikawa, H., Kuroda, M., Ujihara, T., Shuin, T., & Imai, S. (2003). Experimental protection of mice against lethal Staphylococcus aureus infection by novel bacteriophage ΦMR11. Journal of Infectious Diseases, 187(4), 613-624.
37.Merabishvili, M., Pirnay, J. P., Verbeken, G., Chanishvili, N., Tediashvili, M., Lashkhi, N.,Glonti, T., Krylov, V., Mast, J., Parys, l. V., Lavigne, R., Volckaert, G., Mattheus, W., Verween, G., Corte, P. D., Rose, T., Jennes, S., Zizi, M., Vos, D. D., & Vaneechoutte, M. (2009). Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PloS one, 4(3), e4944.
38.Moebus, K. (1987). Ecology of marine bacteriophages. Phage ecology, 137-156.
39.Nakai, T., & Park, S.C. (2002). Bacteriophage therapy of infectious diseases in aquaculture.Res. Microbiol. 153(1), 13-8.
40.Nakai, T., Sugimoto, R., Park, K. H., Matsuoka, S., Mori, K. I., Nishioka, T., & Maruyama, K. (1999). Protective effects of bacteriophage on experimental Lactococcus garvieae infection in yellowtail. Diseases of aquatic organisms, 37(1), 33-41.
41.O'Flynn, G., Ross, R. P., Fitzgerald, G. F., & Coffey, A. (2004). Evaluation of a cocktail of three bacteriophages for biocontrol of Escherichia coli O157: H7.Applied and Environmental Microbiology, 70(6), 3417-3424.
42.Park, S. C., & Nakai, T. (2003). Bacteriophage control of Pseudomonas plecoglossicida infection in ayu Plecoglossus altivelis. Diseases of aquatic organisms, 53(1), 33-39.
43.Perera, R.P., Johnson, S.K., & Lewis, D.H. (1997). Epizootiological aspects of Streptococcus iniae affecting tilapia in Texas. Aquaculture, 152, 25-33.
44.Prasad, Y., Kumar, D., & Sharma, A. K. (2011). Lytic bacteriophages specific to Flavobacterium columnare rescue catfish, Clarias batrachus (Linn.) from columnaris disease. Journal of Environmental Biology, 32, 161-168.
45.Prins, J. M., Van Deventer, S. J., Kuijper, E. J., & Speelman, P. (1994). Clinical relevance of antibiotic-induced endotoxin release. Antimicrobial agents and chemotherapy, 38(6), 1211.
46.Rhoads, D. D., Wolcott, R. D., Kuskowski, M. A., Wolcott, B. M., Ward, L. S., & Sulakvelidze, A. (2009). Bacteriophage therapy of venous leg ulcers in humans: results of a phase I safety trial. Journal of wound care, 18(6), 237-243.
47.Roberts, R.J., & Sommerville, C. (1982). Diseases of tilapia. In: Pullin, R.S.V., Lowe-McConnel, R.H. (Eds.), TheBiology and Culture of Tilapias. International Centre for Living Aquatic Resources Managements, Manila, Philippines, 247– 263.
48.Sanger, F., Nicklen, S., & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences,74(12), 5463-5467.
49.Shanks, R. M., Dashiff, A., Alster, J. S., & Kadouri, D. E. (2012). Isolation and identification of a bacteriocin with antibacterial and antibiofilm activity from Citrobacter freundii. Arch Microbiol. 194(7), 575-587.
50.Sillankorva, S. M., Oliveira, H., & Azeredo, J. (2012). Bacteriophages and their role in food safety. International journal of microbiology, 2012.
51.Stenholm, A. R., Dalsgaard, I., & Middelboe, M. (2008). Isolation and characterization of bacteriophages infecting the fish pathogen Flavobacterium psychrophilum. Applied and environmental microbiology, 74(13), 4070-4078.
52.Sulakvelidze, A., Alavidze, Z., & Morris, J. G. (2001). Bacteriophage therapy. Antimicrobial agents and chemotherapy, 45(3), 649-659.
53.Summers, W. C. (1999). Felix dHerelle and the Origins of Molecular Biology.Yale University Press.
54.Svetlana, J., Dobrila, J. D., & Veljović, LJ. (2003). Citrobacter freundii as a cause of disease in fish. ActaVeterinaria (Beograd), 53(5-6), 399-410.
55.Tsadik, G.G., & Kutty,M.N. (1987). Influence of ambient oxygen on feeding and growth of the tilapia Oreochromisniloticus (Linnaeus). UNDP/FAO/ NIOMR, Port Harcourt, Nigeria, 13 pp.
56.Tung, M. C., Chen, S. C., & Tsai, S. S. (1985). General septicemia of streptococcal infection in cage-cultured Tilapia, Tilapia mossambica, in southern Taiwan. COA Fisheries Series, (4), 95-105.
57.Twort, F. W. (1915). An investigation on the nature of ultra-microscopic viruses. The Lancet, 186(4814), 1241-1243
58.Wang, I. N., Smith, D. L., & Young, R. (2000). Holins: the protein clocks of bacteriophage infections. Annual Reviews in Microbiology, 54(1), 799-825.
59.Wangead, C., Geater, A., & Tansakul, R. (1988). Effects of acid water on survival and growth rate of Nile tilapia (Oreochromis niloticus). In The Second International Symposium on Tilapia in Aquaculture, 15, 433. WorldFish.
60.Watanabe, W. O., Losordo, T. M., Fitzsimmons, k., & Hanley, F. (2002). Tilapia Production Systems in the Americas: Technological Advances, Trends, and Challenges. Reviews in Fisheries Science, 10(3-4), 465-498.
61.Weinbauer, M. G. (2004). Ecology of prokaryotic viruses. FEMS microbiology reviews, 28(2), 127-181.
62.Wiggins, B. A., & Alexander, M. (1985). Minimum bacterial density for bacteriophage replication: implications for significance of bacteriophages in natural ecosystems. Applied and Environmental Microbiology, 49(1), 19-23.
63.Wills, Q. F., Kerrigan, C., &Soothill, J. S. (2005). Experimental bacteriophage protection against Staphylococcus aureus abscesses in a rabbit model. Antimicrobial agents and chemotherapy, 49(3), 1220-1221.
64.Wright, A., Hawkins, C. H., Änggård, E. E., & Harper, D. R. (2009). A controlled clinical trial of a therapeutic bacteriophage preparation in chronic otitis due to antibiotic‐resistant Pseudomonas aeruginosa; a preliminary report of efficacy. Clinical otolaryngology, 34(4), 349-357.
65.Zimmer, M. (2002). Complete genome sequence and characterization of the lysis system of the temperate Clostridium perfringens bacteriophage Φ3626. Institut fürMikrobiologie, ForschungszentrumfürMilch und LebensmittelWeihenstephan (TechnischeUniversitätMünchen), 117.
66.朱鴻鈞、陳葦芋、陳政忻 (2009)。台灣鯛產業概況及趨勢。農業生技產業季刊,19, p.17。
67.行政院農業委員會水產試驗所 (2008)。吳郭魚 168。水產試驗所特刊,10, pp.3-6。
68.李思元, & 莊以光 (2010)。DNA 定序技術之演進與發展。生物醫學暨檢驗科學雜誌,22(2),p.49。
69.彼得. 杜拉克(Peter F. Drucker)著,劉真如譯 (2002)。下一個社會。城邦文化事業股份有限公司,p.24。
70.黃惇聖 (2012)。分離並確認弗氏檸檬酸桿菌為吳郭魚的致病菌 (未出版之碩士論文)。國立嘉義大學農業生物技術研究所,嘉義市。
71.蘇偉成、劉富光 (2005)。台灣水產養殖的永續經營。科學發展,385, pp.45-46。
連結至畢業學校之論文網頁點我開啟連結
註: 此連結為研究生畢業學校所提供,不一定有電子全文可供下載,若連結有誤,請點選上方之〝勘誤回報〞功能,我們會盡快修正,謝謝!
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
系統版面圖檔 系統版面圖檔