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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82878完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳明汝(Ming-Ju Chen) | |
| dc.contributor.author | Chien-Ting Chen | en |
| dc.contributor.author | 陳芊廷 | zh_TW |
| dc.date.accessioned | 2022-11-25T08:01:35Z | - |
| dc.date.copyright | 2021-11-11 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-11 | |
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Effects of antibiotics (oxytetracycline, florfenicol or tulathromycin) on neonatal calves' faecal microbial diversity. Vet. Rec. 177:598. Parameswaran, N., and S. Patial. 2010. Tumor necrosis factor-α signaling in macrophages. Crit. Rev. Eukaryot. Gene Expr. 20:2. Park, H. E., K. H. Do, and W. K. Lee. 2020. The immune-modulating effects of viable Weissella cibaria JW15 on RAW 264.7 macrophage cells. J. Biomed. Res. 34:6. Park, S. Y., G. E. Ji, Y. T. Ko, H. K. Jung, Z. Ustunol, and J. J. Pestka. 1999. Potentiation of hydrogen peroxide, nitric oxide, and cytokine production in RAW 264.7 macrophage cells exposed to human and commercial isolates of Bifidobacterium. Int. J. Food Microbiol. 46:231–241. Parker, D. 1990. Manipulation of the functional activity of gut by dietary and other means (antibiotics/probiotics). J Nutr. 60:639–648. Perdigón, G., M. Locascio, M. Medici, A. P. D. R. Holagado, and G. Oliver. 2003. Interaction of bifidobacteria with the gut and t……… | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82878 | - |
| dc.description.abstract | " 仔牛健康的維持對於酪農產業至關重要，並顯著影響了後續仔牛的生長與生產；而仔牛下痢仍是一種常見的疾病，甚至可影響牧場生產經濟且造成損失。目前多使用抗生素作為治療或預防仔牛下痢的發生，然而由於抗藥性與食品安全的疑慮，許多國家逐漸禁止或減少使用抗生素作為飼料添加劑，因此，研究抗生素之替代物將成為現今相當重要的議題。本研究目的為從健康與下痢荷斯登種仔牛之腸道微生物菌群中，找出關鍵的生物指標菌，作為生物標記，同時分析微生物群間的關聯性和其與下痢之關聯性。此外，本研究將進一步篩選此生物指標菌並透過體外試驗評估其作為益生菌的潛力。 本試驗首先經過專業獸醫師診斷與糞便評分後篩選出健康與下痢仔牛各10頭，並測量其各種血液生化指標與生物標誌物。試驗使用次世代定序法 (NGS) 分析健康組與下痢組中微生物菌相之組成，雖然微生物多樣性分析 (α-diversity) 於兩組間無顯著差異，但是透過微生物樣本比較分析 (β-diversity) 則可將健康及下痢仔牛分群。研究進一步使用線性判別分析 (Linear discriminant analysis effect size, LEfSe) 鑑定出14項菌屬與2項菌種可做為微生物標誌。在下痢組結果中，菌種階層指標菌為bacterium_ic1277，健康組中則鑑定出Bifidobacterium longum subsp. longum作為菌種階層的生物指標菌。其中，菌種B. longum subsp. longum為本試驗健康組微生物標誌物之一，後續運用Spearman相關係數評估發現此菌種與生物標誌物免疫球蛋白G (immunoglobulin G, IgG) 呈現正相關，並與促發炎細胞激素第八型介白素 (Interleukin-8, IL-8) 呈負相關性，顯示B. longum subsp. longum可能具有作為仔牛益生菌的潛力。 因此，本研究進一步以B. longum subsp. longum作為目標，進行菌種的篩選，試驗依序使用了傳統微生物培養法、分子鑑定法與系統分類法，透過16S rRNA基因序列分析並根據neighbor-joining法繪製系統分類樹進行鑑定；結果顯示，經上述菌株鑑定方法後確定7株B. longum subsp. longum。試驗接著以體外試驗評估每個不同B. longum subsp. longum菌株之機能性，包括抗菌活性與免疫條件的功能。在抗菌能力結果中，4株B. longum subsp. longum菌株顯示其具有抑制仔牛下痢病原菌之一沙門氏桿菌 (Salmonella enterica) 的效力。同時，試驗亦選擇促發炎細胞激素腫瘤壞死因子-α (Tumor necrosis factor alpha, TNF-α) 與抑制發炎細胞激素第十型介白素 (Interleukin-10, IL-10) 檢測菌株免疫調節能力。結果指出，B. longum subsp. longum HCF-22和B. longum subsp. longum HCF-27是作為潛力益生菌菌株最適合的選擇。上述二者菌株不僅具有良好的細胞存活率亦具有較佳的免疫調節能力，B. longum subsp. longum HCF-22可抑制S. enterica且具有較低的發炎程度，而B. longum subsp. longum HCF-27則在所有菌株中顯著刺激巨噬細胞分泌最高濃度之IL-10 (P < 0.05)，展現了最佳的抗發炎能力。 綜上所述，本研究透過比較健康和下痢仔牛的腸道微生物菌相以及體外功能益生菌機能性評估，顯示可進行分離和找出用於酪農業的潛在益生菌。而這些發現不僅可進一步的了解仔牛的腸道微生物群，分離出的菌株更可以用於仔牛益生菌飼料添加劑，預防仔牛發生下痢，同時冀望作為抗生素之替代物。 " | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T08:01:35Z (GMT). No. of bitstreams: 1 U0001-0608202113550200.pdf: 5426887 bytes, checksum: 63058d6009e4e6b7be46affa6bb6083d (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "中文摘要 i Abstract iii 目錄 v 圖目錄 ix 表目錄 xi 壹、文獻探討 1 一、仔牛胃腸道 (Gastrointestinal tract, GIT) 環境 1 (一) 仔牛胃腸道組成介紹 1 (二) 仔牛腸道微生物分布 3 (三) 仔牛腸道微生物組功能與其對宿主之交互作用 5 二、仔牛下痢 (Calf diarrhea) 10 (一) 仔牛下痢類型與病徵 10 (二) 仔牛下痢致病機制與致病體種類 11 (三) 仔牛下痢之預防與治療 13 三、仔牛腸道菌相與仔牛下痢之關係 16 (一) 仔牛腸道菌相與仔牛下痢之影響 16 (二) 仔牛腸道菌相與仔牛下痢之潛在機制 17 四、添加活菌於仔牛飼糧之影響 20 (一) 健康狀態、生長性能之影響與作用機制 20 (二) 牛隻腸道微生物相之影響 21 貳、研究動機與目的 28 參、材料與方法 29 第一節：利用微生物體學篩選仔牛腸道中潛力微生物 29 一、試驗流程與實驗設計 29 二、試驗動物 30 三、試驗材料 30 (一) 糞樣 30 (二) 血樣 30 四、試驗方法 30 (一) 健康仔牛與下痢仔牛之現場初篩與試驗分組流程 30 (二) 試驗牛隻生物標誌物 (Biomarker) 與代謝體測定 33 (三) 微生物體學分析 34 (四) 統計分析 43 第二節：潛力菌株之篩選 44 一、試驗流程與實驗設計 44 二、試驗動物 45 三、試驗材料 45 四、試驗方法 45 (一) 目標菌株之篩選 45 (二) 菌株培養與保存方式 48 (三) 目標菌株鑑定 49 第三節：體外試驗 54 一、試驗流程與實驗設計 54 二、試驗方法 55 (一) 目標菌株之特性分析試驗 55 (二) 目標菌株之抗菌活性試驗 (Antimicrobial activity) 55 (三) 菌落計數 59 (四) 候選之乳酸菌株與小鼠巨噬細胞株RAW 264.7共培養 59 (五) 小鼠巨噬細胞株RAW 264.7之細胞存活率試驗 61 (六) 細胞激素測定 62 (七) 統計分析 62 肆、結果 63 第一節：健康與下痢仔牛之腸道微生物體與其關聯性分析 63 一、健康仔牛與下痢仔牛之分組與生物標誌物 (Biomarker) 分析 63 (一) 獸醫專業診斷評判 63 (二) 健康組與下痢組仔牛之血液生化值 67 (三) 血清中蛋白與細胞激素定性試驗 70 二、仔牛腸道微生物體學分析 (Gut microbiome) 72 (一) 腸道微生物DNA萃取 72 (二) 腸道微生物細菌菌相分析 74 (三) 腸道微生物菌相與生物標誌物關聯性分析 103 第二節：自腸道微生物體分析結果篩選仔牛潛在指標菌 109 一、篩選仔牛目標菌株 (生物指標菌) 109 (一) 菌株培養、革蘭氏染色 109 (二) 菌種鑑定 111 第三節：藉由體外試驗測定篩選微生物作為益生菌之特性與功能性 114 一、篩選指標菌之特性分析 114 二、篩選指標菌之功能性分析 117 (一) 指標菌株之抗菌試驗 117 (二) 指標菌株之細胞試驗 123 伍、討論 129 一、仔牛腸道優勢菌群分布 129 二、仔牛下痢對其腸道菌相之影響 132 三、不同健康狀態下仔牛生物標誌物與生物指標菌之關聯性 135 四、不同健康狀態下之腸道菌相功能性 138 五、以生物指標菌作為仔牛益生菌篩選對象 138 六、以功能性試驗探討菌株對病原菌之影響及免疫調節能力 140 (一) 抗菌能力 143 (二) 免疫調節能力 145 陸、結論 150 柒、參考文獻 151 捌、附錄 177" | |
| dc.language.iso | zh-TW | |
| dc.subject | 腸道菌相 | zh_TW |
| dc.subject | 仔牛下痢 | zh_TW |
| dc.subject | 微生物分析 | zh_TW |
| dc.subject | 益生菌 | zh_TW |
| dc.subject | Calf diarrhea | en |
| dc.subject | Gastrointestinal microbiome | en |
| dc.subject | Probiotics | en |
| dc.subject | Microbial analysis | en |
| dc.title | 分析健康與下痢仔牛腸道菌相應用於仔牛益生菌之篩選 | zh_TW |
| dc.title | Analysis of calf’s gastrointestinal microbiome between health and diarrhea for potential probiotic selection | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 莊士德(Shih-Te Chuang) | |
| dc.contributor.oralexamcommittee | 王翰聰(Hsin-Tsai Liu),陳進初(Chih-Yang Tseng),何尚哲 | |
| dc.subject.keyword | 仔牛下痢,腸道菌相,微生物分析,益生菌, | zh_TW |
| dc.subject.keyword | Calf diarrhea,Gastrointestinal microbiome,Microbial analysis,Probiotics, | en |
| dc.relation.page | 177 | |
| dc.identifier.doi | 10.6342/NTU202102149 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2021-08-11 | |
| 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 |
| dc.date.embargo-lift | 2025-08-11 | - |
| 顯示於系所單位： | 動物科學技術學系 | |
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| U0001-0608202113550200.pdf 未授權公開取用 | 5.3 MB | Adobe PDF |
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