臺灣博碩士論文加值系統

本研究是收集分別來自哺餵母乳三個月女嬰的新鮮糞便與喝配方奶的一歲男嬰的新鮮糞便及冷凍母乳，經修飾MRS培養基篩選分離，分別得到30、17及10株菌，先以乳酸菌特性做初步分析確認，結果為：革蘭氏染色陽性菌，觸酶試驗呈陰性反應，厭氧條件下生長，pH值小於4.6，型態為球菌或桿菌；符合上述條件後，挑選分離出之乳酸菌經由API快速菌種鑑定套組分析比對，結果共七株菌種其中2株為Lactobacillus fermentum、1株為Lactobacillus Plantarum、1株為Lactobacillus crisptus、1株為Lactobacillus rhamnosus、1株為Weissella viridescens與1株為Lactococcus lactis ssp. lactis。
經腸道菌數量評估於有益菌分別為：三個月女嬰糞便1.2x109 cfu/g、一歲男嬰糞便2.3x108 cfu/g、冷凍母乳3.9x104 cfu/mL；有害菌則為：三個月女嬰糞便1.3x105 cfu/g、一歲男嬰糞便4.1x105 cfu/g、冷凍母乳未檢測出。

表目錄 vi
圖目錄 vii
中文摘要 viii
英文摘要 ix
壹、前言 1
貳、文獻整理 2
ㄧ、乳酸菌的介紹 2
(一)乳酸菌簡介 2
(二)益生菌定義 2
(三)乳酸菌對腸內菌叢品質之作用機制 3
(四)益生菌應具有的特色 4
(五)乳酸菌作為益生菌使用之條件 6
(六)益生菌保健功效 9
(七)乳酸菌共同特徵 15
二、母乳與配方奶的介紹 15
(一)母乳簡介 16
(一)配方奶簡介 16
三、分離與鑑定方法 18
(一)分類方法 18
(1)表現型之類別與特徵 18
(2)基因型之類別與特徵 19
(二)自動快速鑑定方法 19
(1)Biolog系統 20
(2)Vitek系統 20
參、材料及方法 25
一、實驗材料 26
(ㄧ)樣品採集 26
(二)培養基、緩衝溶液及試劑組成及配製 26
(三)藥品 27
二、實驗設備 27
三、實驗方法 28
(一)培養基配製(修飾MRS Medium) 28
(二)乳酸菌初步分離 28
(三)乳酸菌分離篩選 28
(四)乳酸菌API菌種鑑定 29
(五)菌數測定… 30
(六)菌種保存與活化 30
肆、結果與討論 31
伍、結論 38
陸、參考文獻 39

王西華、王進琦。1996。食品微生物學。pp.17。藝軒圖書出版社。
周建家。2000。益生細菌之篩選及選擇性培養基之建立。中興大學食品科學所碩士論文。
陳勁初。1991。以乳酸菌保存食品之機制。食品工業，23(9)：17-21.
陳慶源、黃崇真、邱雪惠、廖啟成。2007。乳酸菌之保健功效與產品開發。食品生技(11):11-16。
張簡秀楓。2006。乳酸菌的分離與鑑定。中興大學食品暨應用生物科技學所碩士論文。
黃靖堯。2007。台灣酸菜中高胞外多糖益生乳酸菌之篩選與其醃漬益製作酸菜淹蛋可行性之研究。中興大學食品暨應用生物科技學所碩士論文。
程池、楊梅、李金霞、姚粟、胡海蓉。2006。Biolog微生物自動分析系統—細菌鑑定操作規範的研究。食品與發酵工業。32(5): 50-54。
焦玲、程英、田剛。2010。全自動微生物分析系統簡介及應用。檢驗醫學與臨
廖啟成。1998。乳酸菌之分類及應用。食品工業。30(2): 1-10。
廖啟成。2006。乳酸菌之分類及應用。食工所，乳酸菌專輯。3-15。
蔡英傑，1998，乳酸菌與益生菌。生物產業，9(2): 26-32。
Adams,M. R. 1999. Safety of industrial lactic acid bacteria. J. Biotechnol. 68: 171-178.
Beerada, N., Lemeland J. F., G.Laroche, G., Thouventenot, P., and Piaia, Charteris, W.p., Kelly, P.M., Morelli, L., and Collins, J. K. 1998. Development and application of an in vitro methodology to determine the transit tolerance of potenrtially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J. Appl. Microbiol. 84: 759-768.
Carlos Gomez-Gallego, Maria C. Collado, Toni Ilo, Ulla-Marjut JaaKKola, Maria J. Bernal, Maria J. Periago, Seppo Salminen, Gaspar Ros, Rafael Frias. 2012. Infant formula supplemented with polyamines alters the intestinal microbiota in neomatal BALB/cOlaHsd mice. J. Nutr. Biochemistry . 23:1508-1513.
Charteris, W. P., Kelly, P. M., Morelli, L., and Collins, J. K. 1998. Development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J. Appl. Microbiol. 84: 759-768.
Fuller R. Probiotics for farm animals, In Tannock, G W(ed) Robotics：acritical review。Horizon Scientific Press, Norwich. 1999: 15-22.
Fuller, R., Brooker, B.E., 1974.Lactobacilli which attach to the crop epithelium of the fowl.Am.J.Clin.Nutr. 27: 1305-1312.
Gardiner, G. E., O’sullivan, E., Kelly, J., Auty, M. A. E., Fitzgerald, G. F., Collins, J. K., Ross, R. P., and Stanton, C. 2000. Comparative survival ates of human-derived probiotic Lactobacillus paracasei and L.alivarius strains during heat treatment and spray drying. Appl.Environ. Microbiol. 66: 2605-2612.
Havenaar, R., Brink, B.T., and Huis In’t Veld, J. H. J. 1992. Selection of strains for probiotis use in Probiotics, The sciencific basic. 1992. Ed.R.Fuller.Chapman &, Hall London, New York .PP. 209-222.
Hose, H., and Sozzi, T. 1991. Biotechnology group meeting: probiotics-fact or fiction? J. Chem. Technol. Biotechnol. 51: 539-570.
Isolauri. E., Y. Sutas, P. Kankaanpaa, H. Arvilomm, and S. Salminen. 2001. Probiotics: effect on immunity. Am. J. Clin. Nutr. 73(suppl): 444S-50S.
Jonhson, T. R., and Case, C. L. 1998. Laboratory experiment in microbiology. 5th edition. The Benjamin/Cummings Publoshing Co.Inc.
Kandler, O., Weiss, N.1986. Genus Lactobaillus. Page 1209-124 in Bergey’s manual of systematic bacteriology j.g.holt ed. Wilmliams and Wilkin co.Baltimore m. d.
Khedkar, C. D., Mantri, J. M., Garge, R. D., Kulkarni, S. A., and Khedkar, G. D. 1993. Hypocholesterolemic effect of fermentation milks. Cult. Dairy Prod. J. 28: 14-18.
Klaver, F.A.M., and Van Der Meer, R. 1993. The assumed assimilation of cholesterol by lactobacilli and Bifidobacterium is due to their bile salt-deconjugating activity.Appl.Environ.Microbiol. 59(4): 120-1124.
Lilly, D.M.and R. H. Stillwell. 1965.Probiotics：growth promoting factors produced b microorganisms.Science. 147: 47-748.
Lin W. H., Hwang C. F., Chen L. W. , and Tsen H. Y. 2006. Viable counts, characteristic evaluation for commercial lactic acid bacteria products.Food Microbiol .23:4-81.
Lin, M.Y. 1994. The beneficial effects of lactic acid bacteria on human health. J. Chinese. Nutri. Soc. 20:67-380.
M.1991.Bifidobacterium from fermented milks：survival during gastric transit.J.Dairy Sci. 74: 409-413.
MacDonald, L. C., McFeeters, R. F., Daeschel, M. A., and Fleming, H. P. 1987. A defferential medium for the enumeration of homofermentative and heterofermentative lactic acid bacteria. Appl. Environ. Microbiol. 53(6):1382-1384.
Marteau. P., P. SeKsik, and R. Jian. 2002. Probiotics and intestinal health effects: a clinical perspective. Brit. J. Nut. 88(Suppl)1:S51-S57.
Morishita, T.,Tamura, N., Makino, T.,and Kudo, S.1999. Prodution of menaquinones by lactic acid bacteria. J. Dairy Sci. 82:1897-1903.
Mitsuoka, T. 1982. Recent trends in research on intestinal flora. Bifidobacteria Microflora 1: 3-24.
Parvez, S., K. A. Malik, S. A. Kang, and H. Y. Kim. 2006. Probiotics and their fermented food products are beneficial for health. J. Appl Micro. 100: 1171-1185.
Saarela, M.,Lahteenmaki, L., Crittenden, R., Salminen, S., and Mattila –Sandholm, T. 2002. Gut bacteria and health foods-the European perspective.Int.J.Food Microbiol. 78: 99-117.
Salminen, S., Ouwehand, A., Benno, Y., and Lee, Y. K. 1999. Probiotics:how should they be defined? Trends in Food Sci Technol. 10: 107-110.
Salminen, S., von Wrigth, A., Morelli, L., Marteau, P., Brassart, D., Vos de, W. M., Fonde’n, R., Saxelin, M., Collins, K., Mogensen, G., Birkeand, S.E., and Sandholm, T. M., 1998. Demonstration of safety of probiotics -a review. Int. J. Food Microbiol. 44: 93-106.
Sander, M. E., Walker, D. C., Walk, K. M., Aoyama, K., and Klaenhammer, T. R. 1996. Performance of commercial cultures in fluid milk applications. J. Dairy Sci. 76(6):943-955.
Scavage, D. C. 1983. Mechanisms by which indigenous microorganism colonize gastrointestinal epithetial surfaces. Prog. Fd. Nutr. Sci. 7: 65-75.
Shah, N. P.2000b.Effects of milk-derived bioactives: An overview. Br. J. Nutr. 84: S3-S10.
Shah, N. P. 2007. Functional cultures and health benefits. Int. Dairy J. 17: 1262-1277.
Sneath, P. H. A., Mair, N. S., Sharpe, M.E., and Holt, J. G. 1986.Bergey’s manual of systematic bacteriology.Volumn 2. Williams and Wilkins. Baltimore. USA.
Takahashi T., T. Oka, H. Iwana, T. Kuwata, and Yamamoto. 1993. Immune response of mice to orally admintered lactic acid bacteria. Biosci Biotechnol Biochem. 57: 1557-1560.
Uemura, J., T.Itoh, T. Kaneko and K. Noda. 1998. Chemical Characterization of exocellualar polysaccharide from Lactobacillus delbrueckii subsp.bulgaricus OLL1037R-1. Milchwissenschaft 53 (8)：443-446.
Vasiljevic, T., and N. P. Shah. 2008. Probiotics-From Metchnikoff to bioactives. Int. Dairy J. 18(7): 714-728.
Zhou, J. S., Pillidge, C. J., Gopal, P. K., and Gill, H.S.2005.Antibiotic susceptibility profiles of new probiotic Lactobacillus and bifidobacterium strains.Int.J.Food Microbiol. 98：211-217.