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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳明汝 | |
dc.contributor.author | Hsin-Hui Hsieh | en |
dc.contributor.author | 謝馨慧 | zh_TW |
dc.date.accessioned | 2021-06-15T01:30:13Z | - |
dc.date.available | 2016-08-20 | |
dc.date.copyright | 2011-08-20 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-16 | |
dc.identifier.citation | 黃懿儂。2010。台灣黏質發酵乳之機能性研究。國立臺灣大學生物資源暨農學院動物科學技術學系。碩士論文。
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42953 | - |
dc.description.abstract | 本研究目的為探討糖液克弗爾粒於黑糖液、牛乳與羊乳中發酵後之微生物菌相分布,並測試從中分離的乳酸菌株其抗腸炎之機能性。實驗藉由聚合酶連鎖反應–變性梯度膠體電泳及DNA定序等方法鑑定乳酸菌與酵母菌之種類,並以掃描式電子顯微鏡觀察其結構及菌群分布之變化;從糖液克弗爾粒中分離之乳酸菌株,則透過Caco-2細胞單層膜完整性評估,與DSS誘發結腸炎小鼠動物模式探究其保護腸道之能力。
菌種鑑定結果顯示,從不同發酵基質中共分離出6種乳酸菌(Leuconostoc mesenteroides、Lactobacillus mali、Lactobacillus hordei、Lactococcus lactis、Enterococcus faecalis和Bifidobacterium psychraerophilum)與4種酵母菌(Zygosaccharomyces fermentat、Saccharomyces cerevisiae、Dekkera bruxellensis、Pichia fermentans);而直接抽取細菌之DNA可另外發現到Pseudomonas spp.、Enterobacter spp.、Sporolactobacillus spp.和Zymomonas mobilis之存在,以掃描式電子顯微鏡觀察之結果亦發現,不同發酵基質對於糖液克弗爾之菌元結構與微生物菌相分布有顯著影響。 從糖液克弗爾粒中分離出來之Lb. mali可顯著提升Caco-2細胞單層膜之跨上皮電阻值,並增加趨化激素CCL20之分泌量;在DSS誘發結腸炎小鼠動物模式中,亦可減緩因潰瘍性結腸炎而導致之體重減輕、糞便潛血與結腸長度縮短等臨床症狀,組織切片結果同樣應證其對腸道上皮具有保護之功效。 | zh_TW |
dc.description.abstract | Sugary kefir is a fermented beverage, made from sugar and water with or without fruit (usually figs and lemon) added, whose fermentation is induced by grains. Sugary kefir grains are small transparent mucilaginous masses, also consisting of a polysaccharide gel embedding lactic acid bacteria (LAB) and yeasts. In contrast to milk kefir grains made of complex heteropolysaccharides namely kefiran, the sugary kefir grains are mainly consist of dextran, an alpha 1-6 linked glucose polymer. Sugary kefir grains are likely to differ from the milk kefir grains because the materials for fermentation differ widely in their composition. In the present study, we fermented sugary kefir grain in brown sugar, bovine and caprine milk, and evaluated the effect of different fermentation materials on the microbial distribution in both grains and filtrates using culture-dependent and culture-independent methods.
The identification results indicated that the strains in the grains were significantly affected by the fermentation materials. Three LAB species (Leuconostoc mesenteroides, Lactobacillus mali, Lactobacillus hordei) were found in the grains fermented with brown sugar. Whereas, four species including Leu. mesenteroides, Lactococcus lactis, Bifidobacterium psychraerophilum and Enterococcus faecalis were identified in the grains fermented with bovine and caprine milk. For PDA results, three yeasts (Zygosaccharomyces fermentati, Saccharomyces cerevisiae, Dekkera bruxellensis) were found in all three grains. Only Pichia fermentans was identified in the grains grown in the bovine and caprine milk. The profiles obtained by culture-independent denaturing gradient gel electrophoresis (DGGE) generally agreed with the results obtained by culture-dependent methods. However, two additional DGGE bands in the sugary kefir samples were found. Further identification by DNA sequencing revealed that they were Zymomonas mobilis and Sporolactobacillus spp. Additional three bands were also found in the milk samples, they were Pseudomonas spp. and Enterobacter spp. It is worth to notice that the strains and their distributions were different between the grains and filtrates. Additionally, we examined the anti-colitis effects of lactobacilli isolated from sugary kefir grains using in-vitro and on the in-vivo chemical-induced colitis model. Results indicated that Lb. mali could strengthen the epithelial barrier function in vitro by increasing the transepithelial electrical resistance (TEER) with significantly upregulated the level of CCL20 in both apical and basolateral sites. The in-vivo effects of Lb. hordei and Lb. mali on the regulation of intestinal physiology indicated that both strains could ameliorate DSS-induced colitis with a significant attenuation of bleeding score and colon length shortening. In the present study, we demonstrated that the fermentation materials presented remarkable differences in the microbial ecological profiles both in the grains and the filtrates. The strains were varied depending on the carbon and energy sources under grain fermentation. Furthermore, Lb. mali clearly demonstrated an anti-colitis effect. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T01:30:13Z (GMT). No. of bitstreams: 1 ntu-100-R98626007-1.pdf: 5730692 bytes, checksum: c387fc435410ae7b9e4faa3cea987746 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 序言 i
中文摘要 ii 英文摘要 iii 壹、文獻探討 1 一、糖液克弗爾之微生物組成 1 (一)糖液克弗爾 1 (二)菌相分析 2 二、糖液克弗爾之微生物鑑定 5 (一)表現型法 5 (二)逢機增幅多型性DNA 6 (三)限制片段長度多型性 6 (四)聚合酶鏈鎖反應-變性梯度膠體電泳 7 三、糖液克弗爾粒之結構 13 四、糖液克弗爾之機能性 16 貳、材料與方法 19 第一部分:糖液克弗爾粒於不同發酵基質中之微生物菌相分析 19 一、實驗材料 19 (一)糖液克弗爾菌元及其發酵基質 19 (二)微生物培養基 19 二、實驗方法 19 (一)不同發酵基質所需之發酵時間測定 19 (二)糖液克弗爾之製備 20 (三)微生物菌落之分離與保存 20 (四)乳酸菌與酵母菌之DNA純化 21 (五)聚合酶鏈鎖反應 22 (六)變性梯度膠體電泳 24 (七)變性梯度膠體之亮帶純化 25 (八)DNA定序分析 28 (九)分離菌株之生化特性分析 29 (十)掃描式電子顯微鏡觀察 32 第二部分:糖液克弗爾粒分離乳酸菌株抗腸炎機能性之評估 34 一、實驗材料 34 (一)乳酸菌株 34 (二)實驗細胞 34 (三)實驗動物 35 二、實驗方法 35 (一)免疫細胞體外試驗 35 (二)腸道上皮細胞體外試驗 36 (三)結腸炎小鼠動物試驗 38 (四)統計分析 40 叁、結果與討論 41 第一部分:糖液克弗爾粒於不同發酵基質中之微生物菌相分析 41 一、不同發酵基質所需之發酵時間測定 41 二、不同發酵基質之微生物菌相分析 43 (一)由MRS培養基分離之微生物菌相分析 43 (二)由PD培養基分離之微生物菌相分析 53 三、直接抽取樣品DNA之微生物菌相分析 60 (一)以338fGC/518r引子對增幅細菌16S rDNA 60 (二)以NL1GC/LS2引子對增幅真菌26S rDNA. 63 四、掃描式電子顯微鏡觀察 65 第二部分:糖液克弗爾粒分離乳酸菌株抗腸炎機能性之評估 68 一、免疫細胞體外試驗 68 (一)TH1細胞激素 69 (二)抗發炎反應細胞激素 72 (三)前發炎反應細胞激素 74 二、腸道上皮細胞體外試驗 79 (一)單層膜完整性之評估 79 (二)趨化素CCL20之測定 81 三、結腸炎小鼠動物試驗 83 (一)結腸炎臨床症狀之評估 83 (二)組織切片觀察 86 肆、結論 89 參考文獻 90 作者小傳 99 | |
dc.language.iso | zh-TW | |
dc.title | 糖液克弗爾粒於不同發酵基質中之菌相分布與其分離菌株抗腸炎機能性之研究 | zh_TW |
dc.title | Identification of microbial distribution of sugary kefir grains inoculated in different fermented media and investigation of anti-colitis effect of their lactobacilli isolates | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林慶文,黃英豪,陳小玲,陳希嘉 | |
dc.subject.keyword | 糖液克弗爾粒,微生物菌相分布,抗腸炎機能性, | zh_TW |
dc.subject.keyword | Sugary kefir grain,Microbial distribution,Anti-colitis effect, | en |
dc.relation.page | 99 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-16 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
顯示於系所單位: | 動物科學技術學系 |
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