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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳明汝 | zh_TW |
| dc.contributor.advisor | Ming-Ju Chen | en |
| dc.contributor.author | 黃楡茵 | zh_TW |
| dc.contributor.author | Yu-Yin Huang | en |
| dc.date.accessioned | 2024-09-19T16:17:24Z | - |
| dc.date.available | 2024-09-20 | - |
| dc.date.copyright | 2024-09-19 | - |
| dc.date.issued | 2024 | - |
| dc.date.submitted | 2024-08-10 | - |
| dc.identifier.citation | Abouelela, M. E., and Y. A. Helmy. 2024. Next-generation probiotics as novel therapeutics for improving human health: current trends and future perspectives. Microorganisms 12:430. doi: 10.3390/microorganisms12030430
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/95903 | - |
| dc.description.abstract | 傳統益生菌已被廣泛研究和應用，為了開發出新型益生菌，次世代益生菌 (next-generation probiotics) 的概念開始受到重視，次世代益生菌是指「根據微生物群比較分析而鎖定的活體微生物，當其被攝取足量時，可為宿主帶來健康益處」。川崎氏症 (Kawasaki disease) 是一種病因未知、好發於5歲以下兒童的全身性血管炎，伴隨發燒、淋巴結腫大和皮膚黏膜表現等的綜合症。在先前的研究中發現川崎氏症兒童腸道菌相中的Limosilactobacillus mucosae顯著低於健康兒童，顯示L. mucosae可能在川崎氏症中扮演重要角色。因此，本研究旨在從兒童糞便中分離出L. mucosae並研究其特性及功能性，以評估其作為次世代益生菌的潛力。
在本研究共招募到118位健康兒童的糞便樣本，並透過即時聚合酶連鎖反應(real-time polymerase chain reaction, qPCR) 篩選出37個L. mucosae豐度較高的樣本，將其進行細菌分離，共得出16,000個單一獨立菌落，在使用L. mucosae專一性引子對聚合酶連鎖反應 (L. mucosae-specific primer set PCR) 及逢機增幅多型性DNA聚合酶連鎖反應 (randomly amplified polymorphic DNA-PCR) 進行初步鑑定及分型後，透過16S rRNA基因定序、持家基因 (housekeeping gene) pheS及rpoA定序進行進一步的鑑定，最終篩選出兩個相異的L. mucosae菌株，分別命名為H87及H602。 隨後將其和人源的L. mucosae JCM 7735 (R1)、豬源的L. mucosae BCRC 17827T (R2) 及從牛隻糞便所分離之L. mucosae C121、C212、C1988、C251及C10進行菌株特性及功能性之測定。研究結果顯示，低濃度活菌之R1、R2、C1988、C251及H87可以提升促發炎細胞激素腫瘤壞死因子α (tumor necrosis factor-α, TNF-α)，高濃度之熱滅活 (heat-killed) R1、R2、C251及C10能同時提升促發炎細胞激素TNF-α及抗發炎細胞激素白介素-10 (interleukin-10, IL-10)，顯示出這些菌株的免疫調節能力。此外R1、R2及C10在抗氧化試驗中表現最好，具有自由基清除能力、還原力及脂質氧化抑制能力。而抗肥胖能力面，所有菌株皆具有膽鹽水解酶 (bile salt hydrolase) 活性，並且C212、C1988、C10和H87具有膽固醇同化 (cholesterol assimilation) 能力。 綜上所述，本研究從嬰兒糞便中有效的分離出L. mucosae菌株，並和其他來源的L. mucosae菌株相比後，發現H87在免疫調節及降膽固醇方面表現出色，在未來進一步研究其在人體內的功能後，期望能開發出一種新型的潛力益生菌株。 | zh_TW |
| dc.description.abstract | Traditional probiotics have been extensively studied and applied. To develop new probiotics, the concept of next-generation probiotics has begun to gain attention. Next-generation probiotics refer to "live microorganisms identified based on comparative microbiota analyses that, when administered in adequate amounts, confer a health benefit on the host." Kawasaki disease is a systemic vasculitis of unknown etiology that primarily affects children under five years old and is characterized by fever, lymphadenopathy, and mucocutaneous symptoms. Previous studies have found that the gut microbiota of children with Kawasaki disease has significantly lower levels of Limosilactobacillus mucosae compared to healthy children, suggesting that L. mucosae may play an important role in Kawasaki disease. Therefore, this study aimed to isolate L. mucosae from children's feces and investigate its characteristics and functionality to evaluate its potential as a next-generation probiotic.
In this study, fecal samples were collected from 118 healthy children and screened by real-time polymerase chain reaction (PCR) to select samples with high L. mucosae abundance. These samples were then conducted bacteria isolation, and 16,000 single colonies were isolated. After preliminary identification and typing using L. mucosae-specific primers set-PCR and random amplified polymorphic DNA-PCR, further identification was performed using 16S rRNA gene sequencing, housekeeping gene pheS and rpoA sequencing. Finally, two different L. mucosae strains were identified and named H87 and H602. These strains were then compared with L. mucosae JCM 7735 (R1) and L. mucosae BCRC 17827T (R2) isolated from human and pig, and L. mucosae C121, C212, C1988, C251, and C10 isolated from calf feces to determine their characteristics and functionality. The results showed that low doses of live R1, R2, C1988, C251, and H87 could enhance the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α). High doses of heat-inactivated R1, R2, C251, and C10 could enhance both the pro-inflammatory cytokine TNF-α and the anti-inflammatory cytokine interleukin-10 (IL-10), demonstrating the immune regulatory capabilities of these strains. Additionally, R1, R2, and C10 showed the best performance in antioxidant tests, with the ability to scavenge free radicals, reduce power, and inhibit lipid oxidation. In terms of anti-obesity capabilities, all strains exhibited bile salt hydrolase activity, and C212, C1988, C10, and H87 demonstrated cholesterol assimilation ability. In conclusion, this study effectively isolated L. mucosae strains from infant feces. Compared with L. mucosae strains from other sources, H87 showed good performance in immune regulation and cholesterol reduction. After further investigation into its functionality in the human body, the goal of this study is to develop a novel potential probiotic strain. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2024-09-19T16:17:24Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2024-09-19T16:17:24Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 誌謝 i
中文摘要 ii Abstract iv 目次 vii 圖次 x 表次 xii 壹、文獻探討 1 一、 益生菌與次世代益生菌 1 (一) 益生菌 1 (二) 次世代益生菌 6 二、 川崎氏症 (Kawasaki disease) 與Limosilactobacillus mucosae 15 (一) 川崎氏症 15 (二) 川崎氏症與腸道菌相 16 (三) Limosilactobacillus mucosae 16 貳、研究動機與目的 23 參、材料與方法 24 第一節： Limosilactobacillus mucosae之分離與鑑定 24 一、 試驗設計 24 二、 研究材料 25 (一) 受試者招募 25 (二) 糞便樣本收集與前處理 25 三、 研究方法 26 (一) 糞便樣本篩選 26 (二) 菌株分離 (isolation) 與純化 (purification) 27 (三) 菌株鑑定 (identification) 28 第二節： Limosilactobacillus mucosae之特性與功能性測定 43 一、 試驗設計 43 二、 研究材料 44 (一) 試驗菌株 44 (二) 試驗細胞 44 三、 研究方法 46 (一) 菌株特性 46 (二) 菌株功能性 47 肆、結果 57 第一節： Limosilactobacillus mucosae之分離與鑑定 57 一、 受試者招募、糞便樣本篩選及菌落分離 57 二、 菌株鑑定 62 第二節： Limosilactobacillus mucosae之特性與功能性測定 69 一、 菌株特性 69 (一) 菌落外觀 69 (二) 革蘭氏染色 69 (三) 低pH值與膽鹽耐受性 69 二、 菌株功能性 74 (一) Limosilactobacillus mucosae免疫調節能力之評估 74 (二) Limosilactobacillus mucosae對血管發炎之影響 83 (三) Limosilactobacillus mucosae抗氧化能力之評估 85 (四) Limosilactobacillus mucosae抗肥胖能力之評估 88 伍、討論 93 第一節：Limosilactobacillus mucosae之分離與鑑定 93 一、 潛在生物指標之分離與鑑定 93 第二節：Limosilactobacillus mucosae之特性與功能性測定 95 一、 Limosilactobacillus mucosae之菌株特性 95 二、 Limosilactobacillus mucosae之菌株功能性 95 (一) Limosilactobacillus mucosae免疫調節能力之評估 96 (二) Limosilactobacillus mucosae抗氧化能力之評估 97 (三) Limosilactobacillus mucosae抗肥胖能力之評估 98 (四) Limosilactobacillus mucosae之安全性評估 100 陸、結論 101 柒、參考文獻 102 | - |
| dc.language.iso | zh_TW | - |
| dc.title | 潛力益生菌Limosilactobacillus mucosae之分離、鑑定與功能性測定 | zh_TW |
| dc.title | Isolation, identification and functionality test of potential probiotic Limosilactobacillus mucosae | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 112-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 陳勁初;謝睿純;張鑾英;黃麗娜 | zh_TW |
| dc.contributor.oralexamcommittee | Chin-Chu Chen;Jui-Chun Hsieh;Luan-Yin Chang;Lina Huang | en |
| dc.subject.keyword | 次世代益生菌,黏膜乳桿菌,篩菌,免疫調節,降膽固醇, | zh_TW |
| dc.subject.keyword | next-generation probiotics,Limosilactobacillus mucosae,bacteria isolation,immunoregulatory,cholesterol-lowering activity, | en |
| dc.relation.page | 127 | - |
| dc.identifier.doi | 10.6342/NTU202403719 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2024-08-13 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 動物科學技術學系 | - |
| 顯示於系所單位： | 動物科學技術學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-112-2.pdf 目前未授權公開取用 | 4.6 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。