請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/31274
標題: 生物殺菌劑Bacillus subtilis WG6-14作為產蛋雞益生菌及其排遺於植物病害防治應用之評估
The use of biofungicide agent Bacillus subtilis WG6-14 as probiotics in chicken layer rearing and the evaluation of using faecess thereof produced on plant diseases management
作者: 梁瑩如
Liang, Ying-Ru
關鍵字: Bacillus subtilis
γ-polyglutamic acid( PGA)
出版社: 植物病理學系所
引用: 第二章 參考文獻 張德銘,黃勤鎮,林鉅鋃。抗生素濫用影響國人健康專案調查報告。改凱侖出版社/民89。 陳正忻。全球動物生技產業發展現況與趨勢。2007。台灣經濟研究月刊,第 0363 期,p1-17. (available at http://www.biotaiwan.org.tw/download/structure4/ 陳政忻/全球動物生技產業發展現況與趨勢(200712).pdf)。 廖美智,陳正忻。全球動物保健市場暨主要動物生技廠商個案介紹。農業生技產業季刊,2006年第6期,p1-8. (available at http://www.biotaiwan.org.tw/download/structure4/ 陳政忻/第六期-全球動物保健市場暨主要動物生技廠商個案介紹(200606).pdf)。 Aarestrup, FM., Seyfarth, AM., and Emborg, HD. 2001. Effect of abolishment of the use of antimicrobial agents for growth promotion on occurrence of antimicrobial resistance in fecal enterococci from food animals in Denmark. Antimicrob Agents Chemother 45: 2054–2059. Angulo, FJ., Baker, NL., Olsen, SJ., Anderson, A., and Barrett, TJ. 2004. Antimicrobial use in agriculture: controlling the transfer of antimicrobial resistance to humans. Semin Pediatr Infect Dis. 15: 78-85. Caldini, G., Trotta, F. and Cenci, G. 2002. Inhibition of 4-nitroquinolin-1-oxide genotoxicity by Bacillus strains. Res Micriobiol 153: 165–171. Chen X., Chen S., Sun M., and Yu Z. 2005. Optimization by response surface methodology for poly-γ-glutamic acid production using dairy manure as the basis of a solid substrate. Appl Microbiol Biotechnol 69: 390 – 396. Cheng C., Asada Y., and Aida T. 1989. Production of poly-γ-glutamic acid by Bacillus subtilis A35 under denitrifying conditions. Agri Bio Chem 53: 2369–2357. Edward, K., and Arnold, L. 1977. The peptide antibiotics of Bacillus: chemistry, biogenesis, and possible functions. Bacteriol. Rev. 41: 449-474. Faille, C., Membre, J., Kubaczka, M. and Gavini, F. 2002. Altered ability of Bacillus cereus spores to grow under unfavorable conditions (presence of nisin, low temperature, acidic pH, presence of NaCl) following heat treatment during sporulation. J. Food Prot. 65: 1930 – 1936. Fiddaman, P. J., and Rossall, S. 1993. The production of antifungal volatiles by Bacillus subtilis. J. Appl. Bacteriol. 74:119-126. Fuller, R. 1989. Probiotics in man and animals. J. Appl. Bacteriol. 66, 365-378. Glick, BR., and Bashan, Y. 1997. Genetic manipulation of plant growth-promoting bacteria to enhance biocontrol of phytopathogens. Biotechnol. Adv. 15:353-378. Greko, C. 1999. Antibiotics as growth promoters. Acta. Vet. Scand. 92: 87–100 (suppl.). Goto, A., and Kunioka, M. 1992. Biosynthesis and hydrolysis of poly(γ- glutamic acid) from Bacillus subtilis IFO3335. Biosci. Biotech. Biochem. 56: 1031-1035. Hara, T., Fujio, Y., and Ueda, S. 1982. Polyglutamate production by Bacillus subtilis (natto). J. Appl. Biochem. 4: 112- 120. Hara, T., Zhang, JR., and Ueda, S. 1983. Identification of plasmids linked with polyglutamate production in Bacillus subtilis (natto). J. Gen. Appl. Microbiol. 29: 354 – 354. Harrison, JW., and Svec, TA. 1998. The beginning of the end of the antibiotic era? Part I. The problem: abuse of the "miracle drugs". Quintessence Int. 29: 151 – 62. Hezayen, FF, Rehm, BHA., Tindall, BJ, and Steinbϋchel, A. 2001. Transfer of Natrialba asiatica B1T to Natrialba taiwanensis sp. nov. and description of Natrialba aegyptiaca sp. nov., a novel extremely halophilic, aerobic, non-pigmented member of the Archaea from Egypt that produces extracellular poly(glutamic acid). Int. J. Syst. Evol. Microbiol. 51: 1133-42. Hong, HA., Duc, L.H., and Cutting, S.M. 2005. The use of bacterial spore formers as probiotics. FEMS Microbiology Reviews 29: 813 – 835. Impacts of antimicrobial growth promoter termination in Denmark. The WHO International Review panel’s evaluation of the termination of the use of antimicrobial growth promoters in Denmark. Available at http:// www.who.int/salmsurv/en/Expertsreportgrowthpromoterdenmark.pdf Itoh, S. 1997. Alkine cellulases from alkaliphilic Bacillus: enzymeproperties, genetics and applications to detergents. Extremophiles, 1: 61 – 66. Ivánovics, G., and Bruckner, V. 1937a. Chemical and immunologic studies on the mechanism of anthrax infection and immunity. I. The chemical structure of capsule substance of anthrax bacilli and its identity with that of the B. mesentericus. Z. Immunitaetsforsch. 90: 304–318. Ivanovics, G., and Bruckner, V. 1937b. The chemical nature of the immuno-specific capsule substance of anthrax bacillus. Naturwissenschaften. 25: 250. Ivánovics, G, and Erdös, L. 1937. Ein Beitrag zum Wesen der Kapsel-substanz des Milzbrandbazillus. Z. Immunitätsforsch 90: 5-19 Kunioka, M. 1997. Biosynthesis and chemical reactions of poly(amino acid)s from microorganism. Appl. Microbiol. Biotechnol. 47: 469 – 475. Kyriakis, SC., Tsiloyiannis, VS., Vlemmas, J., Sarris, K., Tsinas, AC., Alexopoulos, C., and Jansegers, L. 1999. The effect of probiotic LSP122 on the control of post-weaning diarrhea syndrome in piglets. Res. Vet. Sci. 67: 223 – 228. LaTagione, RM., and Woodward, MJ. 2003. Competitive exclusion by Bacillus subtilis spore of Salmonella enterica serotype Enteritidis and Clostridium perfringens in young chickens. Vet. Microbiol. 94: 245-256. Martin, O. and Alexander, S. 2004. Microbial Degradation of Poly(amino acid)s. Biomacromolecules 5: 1166 – 1176. Mazza, G. 1983. Genetic studies on the transfer of antibiotic resistance genes in Bacillus subtilis strains. Chemioterapia. 2: 64 – 72. Mazza, P. The use of Bacillus subtilis as an anti-diarrhaeal microorganism. 1994. Boll. Chim. Farmceutico. 133: 3 -8. Mellon, M., Benbrook, C., and Benbrook, K. 2001. Hogging it: estimates of antimicrobial abuse in livestock. Cambridge, Union of Concerned Scientists Publications. Microbial threats to health: Emergence, detection, and response. 2003. Report of the Institute of Medicine. Available at http://www.nap.edu/books/030908864X/html Moir, A., and Smith, DA. 1990. The genetics of bacterial spore germination. Annu. Rev. Microbiol. 44: 531–553. Nguyen, KM. Tam, Nguyen Q., Uyen, HA., Hong, LeH., Duc, Tran TH., Claudia RS,, Adriano OH., and Simon, MC. 2006. The intestinal life cycle of Bacillus subtilis and close relatives. J. Bacteriol. 188: 2692–2700. Niemetz, R., Kärcher, U., Kandlera, O., Tindall, BJ., and König, H. 1997. The cell wall polymer of the extremely halophilic archaeon Natronococcus occultus. Eur J Biochem. 249: 905-11. Omeira, N., Barbour, EK., Nehme, PA., Hamadeh, SK., Zurayk, R., and Bashou, I. 2006. Microbiological and chemical properties of litter from different chicken types and production systems. Sci.Total Environ. 367: 156–162. Pötter, M., Oppermann-Sanio, FB., and Steinbüchel, A. 2001. Cultivation of bacteria producing polyamino acids with liquid manure as carbon and nitrogen source. Appl. Environ. Microbiol. p. 617–622. Report on the proposal for a European Parliament and Council Regulation on additives for use in animal nutrition. 2006. Document No.: A5-0373/2002. Available at http://www.health.fgov.be/WHI3/krant/krantarch2002/kranttekstnov2/021114m13eu.htm Rhee, KJ., Sethupathi, P., Friks, A., Lanning, DK., and Knight, KL. 2004. Role of commensal bacteria in development of gut-associated lymphoid tissues and preimmune antibody repertoire. J. Immunol. 172: 1118–1124. SCAN. Report of the Scientific Committee on Animal Nutrition on product BioPlus 2B® for use as feed additive. European Commission, Health and Consumer Protection Directorate - General. (SCAN) Scientific Committee on Animal Nutrition. 2000. Available at http://europa.eu.int/comm/food/fs/sc/scan/out49_en.pdf. SCAN. Assessment by the Scientific committee on Animal Nutrition (SCAN) of a microorganism product: Esporafeed Plus®. European Commission, Health and Consumer Protection Directorate - General. (SCAN) Scientific Committee on Animal Nutrition. 1999. Available at http://europa.eu.int/comm/food/fs/sc/scan/out39_en.pdf. Schallmey M, Singh A, and Ward OP. 2004. Developments in the use of Bacillus species for industrial production. Can J Microbiol. 50: 1-17. Shea, K.M. 2003. Antibiotic resistance: What is the impact of agricultural uses of antibiotics on children’s health? Pediatrics 112: 253 – 258. Shin, IL., and Van YT. 2001. The production of poly–(γ - glutamic acid) from microorganisms and its various applications. Bioresour. Technol. 79: 207–225. Sung, MH., Park, C., Kim,CJ., Poo,H., Soda, K. and Ashiuchi, M. 2005. Natural and edible biopolymer poly-γ-glutanic acid: synthesis, production and applications. The Chemical Record 5: 352 – 366. Sorensen, TL., Wegener, HC., and Frimodt-Moller, N. 2002. Resistant bacteria in retail meats and antimicrobial use in animals. N Engl J Med 346: 777 – 779 (letter). Tannock, GW. Modification of the normal microbiota by diet, stress, antimicrobial agents and probiotics. In: Mackie, R.I., With, B. A., Isaacson, R.E. (Eds.), Gastrointestinal Microbiology, Vol. 2, Gastrointestinal Microbes and Host Interactions. Chapman and Hall Microbiology Series, International Thomson Publishing, pp. 434 – 465. van den Bogaard AE., and Stobberingh, EE. 1999. Antibiotic usage in animals: impact on bacterial resistance and public health. Drugs. 58: 589-607. Verschuere, L., Rombaut, G., Sorgeloos, P. and Verstraete, W. 2000. Probiotic bacteria as biological control agents in aquaculture. Mic. Mol. Biol. Rev. 64: 655-671. Wierup, M. 1998. Preventive methods replace antibiotic growth promoters: ten years experience from Sweden. APUA Newsletter 16: 1-4. Whipps, JM. 2001. Microbial interactions and biocontrol in the rhizosphere. J. Exp. Bot. 52:487-511. World Health Organization. 1997. The medical impact of the use of antimicrobials in food animals: report and proceedings of a WHO meeting, Berlin, Germany, October 13–17. World Health Organization. 1999. Containing Antimicrobial Resistance: Review of the literature and report of a WHO workshop on the development of a global strategy for the containment of antimicrobial resistance. Geneva, Switzerland, February 4 – 5. 參考文獻 劉孟宗、施宗雄。2002。蛋雞糞添加特定微生物對堆肥化影響之研究。畜產研究 35: 205 - 213。 夏良宙。1996。雞舍空氣中之不良因素及其影響。中國畜牧雜誌第五十五冊合訂本第 28 卷第 4 期 p. 89 ~ 96。 Abdulrahim SM, Haddadin SY, Hashlamoun EA, and Robinson RK. 1996. The influence of Lactobacillus acidophilus and bacitracin on layer performance of chickens and cholesterol content of plasma and egg yolk. Br. Poult. Sci. 37:341-346. Bacon CW. 1986. Effects of broiler litter volatiles and ammonia on fungal spore germination. Poult Sci. 65: 710-6. Biochemical Tests for Identification of Medical Bacteria. 2000. Chap. 12. Gelatin Liquefaction Tests, pp. 174-175. Chen X., Chen S., Sun M., Yu Z. Mediu. 2005. Optimization by response surface methodology for poly-γ-glutamic acid production using dairy manure as the basis of a solid substrate. Appl Microbiol Biotechnol 69: 390 – 396. Gershanovich VN, Bol''shakova TN, Dobrynina OIu, Galushkina ZM, Kukanova AIa, Stepanov AI. 2005. Nitrogen assimilation enzymes in Bacillus subtilis mutants with hyperproduction of riboflavin. Mol Gen Mikrobiol Virusol: 29-34. Guo X, Li D, Lu W, Piao X and Chen X. 2006. Screening of Bacillus strains as potential probiotics and subsequent confirmation of the in vivo effectiveness of Bacillus subtilis MA139 in pigs. Antonie Van Leeuwenhoek. 90:139-46. Haddadin MS., Abdulrahim SM., Hashlamoun EA., and Robinson RK. 1996. The effect of Lactobacillus acidophilus on the production and chemical composition of hen’s eggs. Poult. Sci. 75: 491-494. Hoffmann, T., N. Frankenberg, M. Marino and D. Jahn. 1998. Ammonificationin Bacillus subtilis utilizing dissimilatory nitrite reductase is dependent on resDE. J. Bacteriol. 180:186–189. Hong HA, Duc L.H., Cutting S.M. 2005. The use of bacterial spore formers as probiotics. FEMS Microbiology Reviews 29: 813 – 835. Hosoi T, Ametani A, Kiuchi K, Kaminogawa S. 1999. Changes in fecal microflora induced by intubation of mice with Bacillus subtilis (natto) spores are dependent upon dietary components. Can J Microbiol. 45: 59-66. Itoh S. 1997. Alkine cellulases from alkaliphilic Bacillus: enzymeproperties, genetics and applications to detergents. Extremophiles 1: 61 – 66. Jin LZ, Ho YW, Abdullah N, and Jalaludin S. 1998. Growth performance, intestinal microbial populations and serum cholesterol of broilers diets containing Lactobacillus cultures. Poult. Sci. 77:1259-1265. Kithome, M., Paul, JW., and Bomke, AA., 1999. Reducing nitrogen losses during simulated composting of poultry manure using adsorbents or chemical amendments. Journal of Environmental Quality. 28: 194-201. Lalloo R, Ramchuran S, Ramduth D, Görgens J, Gardiner N. 2007. Isolation and selection of Bacillus spp. as potential biological agents for enhancement of water quality in culture of ornamental fish. J Appl Microbiol.103: 1471-9. Leonard CG., Housewright RD., and Thorne CB. 1958. Effects of some metallic ions on glutamyl polypeptide synthesis by Bacillus subtilis. J. Bacteriol. 76: 499 - 503. Linda C., Douglas and Mary E. Sanders. 2008. Probiotics and prebiotics in dietetics practice. J Am Diet Assoc. 108: 510-21. Mohan B, Kadirvel R, Bhaskaran M, and Natarajan A. 1995. An Effect of probiotic supplementation on serum/yolk cholesterol and on egg shell thickness in layers. Br. Poult. Sci. 36:799-803. Ping HU, Terrance Leighton, Galina Ishkhanova and Sydney Kustu. 1999. Sensing of Nitrogen Limitation by Bacillus subtilis: Comparison to Enteric Bacteria. Journal of Bacteriology, p. 5042–5050 Pötter M, Oppermann-Sanio FB, Steinbüchel A. 2001. Cultivation of bacteria producing polyamino acids with liquid manure as carbon and nitrogen source. Applied and Environmental Microbiology, Feb, p. 617 – 622. Ogunwande GA, Osunade JA, Adekalu KO, and Ogunjimi LA. 2008. Nitrogen loss in chicken litter compost as affected by carbon to nitrogen ratio and turning frequency. Bioresour Technol. 99: 7495-503. Omeira N., Barbour E.K., Nehme P.A., Hamadeh S.K., Zurayk R., Bashou I. 2006. Microbiological and chemical properties of litter from different chicken types and production systems. Science of the Total Environment 367, 156 - 162. Shih. IL., Yu, YT. 2005. Simultaneous and selective production of levan and poly(γ-glutamic acid) by Bacillus subtilis. Biotechnology Letters 27: 103 - 106. Teresa M. Barbosa, Cla′udia R. Serra, Roberto M. La Ragione, Martin J. Woodward, and Adriano O. Henriques. 2005. Screening for Bacillus Isolates in the Broiler Gastrointestinal Tract. Applied and Environmental Microbiology, p. 968–978. Weatherburn M.W. 1967. Phenol – Hydrochloride reaction for determination of ammonia. Anal. Chem. 39 (8): 971 - 974. Xiong Chen, Shouwen Chen, Ming Sun, Ziniu Yu. 2005. High yield of poly-γ-glutamic acid from Bacillus subtilis by solid-state fermentation using swine manure as the basis of a solid substrate. Bioresource Technology 96: 1872 - 1879. Xiong Chen, Shouwen Chen, Ming Sun, Ziniu Yu. 2005. Medium optimization by response surface methodology for poly-γ-glutamic acid production using dairy manure as the basis of a solid substrate. Appl Microbiol Biotechnol 69: 390 - 396. Yurong Y, Ruiping S, Shimin Z, Yibao J. 2005. Effect of probiotics on intestinal mucosal immunity and ultrastructure of cecal tonsils of chickens. Arch Anim Nutr. 59: 237-46.
摘要: Bacillus subtilis WG6-14 為台灣本土性可產生內生孢子之菌株,其原始分離自根圈土壤且具有廣效性抗真菌及細菌之功效。目前在生物殺菌劑之應用已相當成功,且商品化產品已成功應用在植物病害防治。此外,關於 Bacillus subtilis 亦已廣泛應用在動物飼養上,且展現益生菌之功效。本研究主要目的在於評估 Bacillus subtilis WG6-14 作為產蛋雞用益生菌之可能性。本研究將受試之 B. subtilis WG6-14 分別添加在飼料及飲水中,其中飼料試驗者係包括 30 隻 50 週齡之試驗雞隻,與 30 隻 50 週齡之對照雞隻;飲水試驗者係包括 600 隻 50 週齡之試驗雞隻,與 150 隻 50 週齡之對照雞隻。結果發現添加在飲水試驗中之試驗組,其飲水中多為典型 B. Subtilis WG6-14 菌落,而少有其他型態菌落出現;但對照組之飲水中則總菌量較高,且咸為枯草桿菌以外成員,待鑑定之微生物種類相當複雜。而將 WG6-14 添加於飲水中之試驗組,其糞便中之Salmonella 與 Staphylococcus 之微生物數量明顯較對照組低 1-2 個對數值,而 coliform 數量則明顯較對照組低約 1 個對數值。此外,在膽固醇部分,飲水中含有適量的 WG6-14 之試驗組,其蛋中膽固醇濃度亦較對照組降低約 24% 。本研究所見 WG 6-14 添加應用下,對於雞隻飼育上之諸多助益效果,顯示其於飼料添加上之應用性值得重視。 本研究進一步收集前述受試雞隻與對照雞隻之糞便排遺,以評估其應用在植物生長促進與植物病害防治之應用性。結果發現,未經堆肥化處理之雞糞,在應用上明顯可造成甘藍種子發芽不佳之影響,唯此一負面效果於 WG6-14 飼育下所收集到含有 WG6-14 之雞糞,在應用上較不明顯,且其處理植株之生長勢以葉寬受測結果看顯有比對照組為佳之現象。此外,本研究進一步發現,在雞糞混拌後,如先經數天堆置,則在小葉百喜草誘釣測試已可證實其可明顯降低土壤中存在的 R. solani AG4 之活力,此一對 R. Solani 之抑制效果在混拌含 WG6-14 雞糞 3 天即可見之而在利用未含有 WG6-14 之雞糞添加處理組,則於堆置 3 天後亦可見 AG4 活力降低的現象,然與含 WG6-14 孢子處理組相較,則有明顯差異,此一結果明白顯示堆置雞糞中含有適量 WG6-14 的存在,對於土中病原菌數量之降低確有正面效益。Bacillus 屬菌株已知可產生聚麩胺酸 (poly(γ-D-glutamic acid)(PGA))等多種聚氨基酸。針對聚麩胺酸所具有之優異保水功效一特質,本研究嘗試篩選可產生高量聚麩胺酸及兼具動物用益生菌應用性之菌株,以期能在供動物使用後,可進一步應用在植物生長及病害防治等方面。結果顯示,γ- PGA 之產生特性與對動物病原之抗生活性不見得有關,或可配合不同使用目的選取已知具不同功能之益生菌混合應用,其應可較單一菌株供應可獲較佳之成果。
Bacillus subtilis WG6-14, a Taiwan-native endospore forming strain obtained from guava rhizosphere, has been shown to be antagonistic against wide spectrum important phytopathogenic fungi and bacteria. The development of its biofungicide application has been successful, its commercialized use in plant disease control is now in progress. As B. subtilis has been widely applied as direct-fed microbial (DFM) known with beneficial bioregulator function in animal science, the possible use of B. subtilis WG6-14 as probiotic in chicken layer rearing was explored. Both liquid and powder formulations of the tester strain (contains approximately 1010 endospores per ml/gm) produced by a fermentation pilot plant located at the campus were used for the conducted tests. The tester DFM was applied as supplements together with either the chicken feeds or water supply at final concentration approximately 108 endospores per ml/gm. A total of 30 heads (trial with powder formulation) and 750 heads (trial with liquid formulation) of chicken layers at 50 wks old were used for the tests. In liquid formulation applied trial, the fermentor produced broth culture of WG6-14 was administered to drinking water to 600 heads, the rest 150 heads without WG6-14 supplementation in drinking water were used as compared control. The daily egg yield and quality were monitored on a weekly basis. A paralleled analysis was shown by dilution plate count the micro flora within the drinking water and the layer little. The result obtained indicated the presence of solely WG6-14 in drinking water among the treated groups, the presence of contaminant microbes appeared to be non-detectable. In contrast to this, that among the non-treated group contained various kinds of contaminant microorganisms. Also worth noting was that the layer faeces from the control group consisted of bountiful amount of coliform bacteria and presence of substantial amount of Salmonella species and Staphylococcus species. Whereas the layer faeces collected from WG6-14 treated group contains approximately 104 cfu/gm of WG6-14, no Salmonella and a very much reduced numbers of coli-form and Staphylococcus spp. bacteria. Results obtained 5 weeks after treatment indicated a substantial decrease among WG6-14 treated layers the cholesterol content of egg yolks by approximately 24% as compared to the non-treated control. The observed beneficial effect of WG6-14 indicated its usefulness as probiotic feed additives to layer rearing.The chicken faecess collected from the experimental farm were evaluated further for plant growth promotion and the potential of plant disease control application. The amendment of collected chicken faeces obtained from the WG6-14 treaded birds resulted in certain growth promoting effect on cabbage foliage development, although the rate a test soil (artificially inoculated with Rhizoctonia solani AG4) with seedling emergence was affected. The compost chicken faeces collected from WG6-14 fed birds was found effective in reducing the inoculums potential of AG4. Bioassay by bahia-grass baiting indicated a significant reduction of R. solani AG4 inoculum potential by composting with 2% faeces amendment. It was worth noting that the decrease AG4 propagules was consistently greater in chicken faeces with WG6-14 comparing to that without WG6-14 amendment.Furthermore, Bacillus spp. are gram-positive bacteria, and are able to synthesize the polyamino acid (PAA), poly(γ-D-glutamic acid)(PGA) as a capsular substance or as a water-soluble slime. PGA is an excellent water-keeping material. The potential of Bacillus spp. for producing γ-PGA and the relationship of γ-PGA producing ability and the probiotic function were explored. Results obtained indicated the productivity of γ-PGA of a test strain did not correlate well the effectiveness of probiotic function. For the intended probiotics application, the use of mixed strains each with suitable desired function may be a more appropriated way than the application of a single strain.
URI: http://hdl.handle.net/11455/31274
其他識別: U0005-2308200814592500
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