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类型抑制幽門桿菌發酵乳之研究.doc

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    抑制幽門桿菌發酵乳之研究.doc
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    1、抑制幽門桿菌發酵乳之研究Study on the fermented milk for inhibit Helicobacter pylori研究生:陳義昌 Chen Yi-Chang指導教授:謝寶全 Xie Bao-Quan【摘要】本研究收集市售商品及自然界樣品分離出生長快速之乳酸菌,並測試對幽門桿菌的抑制能力,挑選出兩株不同來源的乳酸菌,經菌種鑑定後,認定分離自市售發酵乳的 2 號菌株為Lactobacillus paracasei;分離自醃漬蔬菜的 26 號菌株為 Lactobacillus plantarum,兩株乳酸菌發酵牛乳之後能在培養皿上抑制幽門桿菌 BCRC 17023 和 B

    2、CRC 17027,產生 17 - 22 mm 的抑菌圈。L. paracasei 菌株和產生的乳酸為抑制幽門桿菌的主要原因,在發酵乳中能夠抑制幽門桿菌產生明顯的抑菌圈,而發酵乳上清液(FMS)在體外 2 個小時能抑制 45%的幽門桿菌,並降低幽門桿菌所產生的尿素酶活性 56 %;L. plantarum 能分泌出某種抑菌物質,在中性 FMS 中 2 個小時能抑制 33 - 37%的幽門桿菌,並降低 40 - 60%的尿素酶活性,乳酸存在的低 pH 值的條件下,FMS 可以在 1個小時內殺死試管中所有的幽門桿菌(8.8 Log CFU/mL)。但是兩株乳酸菌的發酵乳經過 100,30分鐘殺菌後

    3、則失去對幽門桿菌的抑制能力。動物試驗中,兩株乳酸菌的發酵乳讓小鼠連續飲用 3週,能減少小鼠胃中之幽門桿菌數量約 3 個對數值,並相對抑制尿素酶活性反應。顯示本研究之乳酸菌可以預防幽門桿菌感染。在乳酸菌的安全性上,ICR、C57BL/6J 兩個品系的小鼠飲用發酵乳 6 - 8 週後,都沒有發生生理上的明顯差異及病變。在發酵乳的應用上,L. paracasei 對脫脂乳的發酵性較好,可以直接發酵脫脂乳,但無法忍受消化道環境之低 pH 值(pH 3.0)和高濃度 0.3%膽鹽;L. plantarum 需要添加 0.5%以上的 yeast extract 於脫脂乳中,才可以得到較好的發酵性,而此菌對

    4、消化道環境適應性良好,可以存活於腸胃中。兩株乳酸菌在共同生長時不會有互相抑制的情形,並在製成發酵乳後儲藏於 4中 60 天後,仍可保有活菌數 7.05 Log CFU/mL。綜合上述,本研究所使用的兩株乳酸菌可以預防幽門桿菌感染,在食用上為安全無虞,可以用於製造發酵乳並具有良好的儲藏性。關鍵字:乳酸菌、抑制、幽門桿菌【Abstract 】Isolated strains of lactic acid bacteria (LAB) from commercial production and natural sources were tested for their efficiency in

    5、inhibiting Helicobacter pylori in this study. The No.2 strain of Lactobacillus paracasei was isolated from commercial fermented milk and the NO.26 strain of Lactobacillus plantarum was isolated from pickled vegetables. Two LAB strains could inhibited H. pylori BCRC 17023 and BCRC 17027 on the fermen

    6、ted milk plate and produced the inhibitory zones were 17 to 22 mm. The main reasons of inhibition H. pylori were alive bacteria and produced lactic acid by L. paracasei that was incubated with the fermented milk could produce obvious inhibitory zone. In vitro test, fermented milk supernatant (FMS) o

    7、f L. paracasei could kill 45% H. pylori and decreased 56% the urease activity in 2 hours. In addition, L. plantarum could excrete certain bacteriocins. In vitro test, neutral fermented milk supernatant (NFMS) of L. plantarum could kill 33 to 37% H. pylori and decreased 40 to 60% the urease activity

    8、in 2 hours. Using FMS would be kill the H. pylori (8.8 Log CFU/mL) for 100% in 1 hour. But two LAB strains of FMS after, 30 min sterilized 100 lost the ability to inhibit H. pylori. In vivo, mice drank fermented milk for four week, the results showed that the H. pylori decreased to approximately 3 l

    9、og cfu/ml and inhibited urease activity reaction in stomach. On the security of the LAB, after the mice of ICR and C57BL/6J two strains drank fermented milk 6 to 8 weeks, there were no obvious difference in the occurrence of physiological and pathological change. Application of fermented milk, the f

    10、erment of L. paracasei was better in the skim milk and could directly fermented the skim milk, but unable to tolerate low pH value 3.0 and high concentration of 0.3% bile salts in the environmental of alimentary canal. When skim milk added more than 0.5% the yeast extract, L. plantarum were got bett

    11、er fermentation, survivability and adapted fine in the environmental of alimentary canal. While two LAB strains each other growth will be no inhibition condition and made fermented milk was 7.05 Log CFU/mL of alive LAB in store 60 days at 4. The results showed that fermented milk in this research can prevent the infecting from H. pylori and do not have any toxic for the security in eating, may be used for making fermented milk and possess good shelf life. Key words: lactic acid bacteria, inhibit, Helicobacter pylori

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