臺灣博碩士論文加值系統

近年來由於抗生素的過度使用造成許多菌種產生抗藥性，因此尋找具有抗菌能力的天然抗菌物質成為現在極度受到重視的問題。本研究分別測試中草藥及真菌代謝物的抗菌能力，並採用真菌及中草藥的合併發酵進行研究，觀察發酵後效果變化判斷其作為抗菌劑的潛力。
本研究使用牛樟芝生產的發酵液使用於實驗，並選用六種不同的中草藥(龍眼殼、龍眼核、刺蔥莖部、仙人掌、左手香、金銀花)分別經過酒精萃取(95%、75%、50%)以及熱水萃取(80℃、121℃)
，並測試其抑菌效果後作為發酵添加劑與牛樟芝進行合併發酵，並以常見的兩種感染病原菌金黃葡萄球菌(Staphylococcus aureus)與大腸桿菌(Escherichia coli)以及植物病原菌青枯雷爾氏菌(Ralstonia solanacearum)進行實驗。結果發現牛樟芝發酵液與六種中草藥的酒精萃取液250 mg/ml對金黃葡萄球菌以及大腸桿菌皆有抑菌圈生成，而熱水萃取液則無明顯效果出現。對青枯菌則皆無明顯效果產生。
實驗結果表示牛樟芝液態發酵與固態發酵在相近的發酵天數下，以液態發酵效果較佳。而牛樟芝發酵液對大腸桿菌、金黃葡萄球菌與青枯菌的最小抑菌濃度(minimum inhibitory concentration :MIC)分別為7.8 mg/ml、1.95 mg/ml與7.8 mg/ml，與最小殺菌濃度(minimum bactericidal concentration :MBC)為15.6 mg/ml、1.95 mg/ml與7.8 mg/ml已有確定具有明確的抗菌能力。
本研究結果更證明牛樟芝與中草藥合併發酵能提升牛樟芝抗菌效果，其中又以牛樟芝與龍眼核75 %酒萃物合併發酵效果對大腸桿菌及青枯菌最好，抑菌圈分別為29.00 ±0.57 mm與21.33 ±0.88 mm，對金黃色葡萄球菌則是與刺蔥莖95 %酒萃物發酵效果較好抑菌圈為31.67 ±0.58 mm的抑菌圈，這表示添加不同中草藥進行發酵可改變對不同菌種的抑制效果，因此牛樟芝合併中草藥發酵具有成為天然抗菌劑的潛力。

In recent years, the wide use of antibiotics lead to the emergence of multidrug resistant bacterial strains, therefore it is essential to discover new natural substances from microbial sources with antibacterial activities. In this study ,we tested various herbs, fungal metabolites, and fermented herbs with fungi to analysis their antimicrobial activities.
The results of this study show herbs Dimocarpus longan seed, Dimocarpus longan pericarp ,Zanthoxylum ailanthoides stecn , Cactaceae, Coleus ambolnicus Lour and Lonicera japonica Thunb , extracted by various Concentrations of alcohol or different temperature of water, fermented by Antrodia can enhance the antibacterial effect.
The results showed that liquid fermentation and solid-state fermentation of Antrodia Camphorata in the similar fermentation days ,Liquid fermentation is more efficient than solid-state fermentation. cultivating 21 days fermentation broth of Antrodia camphorate to E. coli, Staphylococcus aureus and Ralstonia solanacearum ,the minimum inhibitory concentration were 7.8 mg/ml,1.95 mg/ml and 7.8 mg /ml, the minimum bactericidal concentration were 15.6 mg / ml, 1.95 mg/ml and 7.8 mg/ml.
Among them fermented Dimocarpus longan extracted by 75% alcohol was the best of Escherichia coli and Ralstonia solanacearum . Inhibition zone was 29.00 ±0.57 mm and 21.33 ±0.88 mm. Them fermented Dimocarpus longan extracted by 95% alcohol was the best of Staphylococcus aureus. Inhibition zone was 31.67 ±0.58 mm. Therefore, herbal fermented by Antrodia possess potentials as a natural antibacterial agent.

中文摘要 i
英文摘要 ii
致謝 iii
目次 iv
圖附錄 vii
表附錄 ix
第一章 前言 1
第二章 文獻回顧 3
2.1抗生素與抗藥性 3
2.1.1 抗生素 3
2.1.2 抗藥性 4
2.2菌種簡介 4
2.2.1金黃色葡萄球菌簡介 4
2.2.2大腸桿菌簡介 5
2.2.3青枯菌簡介 6
青枯病的防治現狀 7
2.2.4牛樟芝簡介 8
牛樟芝的型態與命名 9
牛樟芝現有栽培法 10
牛樟芝相關研究 11
2.3中草藥簡介 13
2.3.1龍眼簡介 13
2.3.2刺蔥簡介 14
2.3.3仙人掌簡介 14
2.3.4左手香簡介 15
2.3.5金銀花簡介 16
第三章 研究目的 8
第四章 材料與方法 19
4.1實驗架構 19
4.2材料 20
4.2.1菌種 20
4.2.2中草藥 20
4.2.3器材 20
4.2.4藥品 21
4.2.5培養基配方 21
4.2.6樣品代號 22
4.3實驗方法 24
4.3.1菌種培養、保存 24
4.3.1.1菌種培養 24
4.3.1.2菌種保存 24
4.3.2中草藥萃取樣品製備 24
4.3.2.1中草藥酒精萃取 24
4.3.2.2中草藥熱水萃取 25
4.3.3牛樟芝發酵 25
4.3.3.1牛樟芝液態發酵 25
4.3.3.2牛樟芝固態發酵 25
4.3.3.3牛樟芝與中草藥合併液態發酵 25
4.3.3.4牛樟芝液態發酵耐熱測試 26
4.3.4抑菌實驗 26
4.3.4.1紙碇擴散法 26
4.3.3.2 MIC測試 27
4.3.3.3 MBC測試 27
4.3.5 HPLC分析 28
第五章 結果與討論 29
5.1中草藥抑菌能力評估 29
5.1.1中草藥紙碇擴散實驗 29
5.1.2中草藥最小抑菌濃度與最小殺菌濃度 31
5.2樟芝抑菌能力評估 49
5.2.1牛樟芝液態發酵與固態發酵抑菌功能評估 49
5.3牛樟芝與中草藥合併發酵抑菌能力評估 56
5.3.1牛樟芝合併發酵對大腸桿菌的抑制能力評估 56
5.3.2牛樟芝合併發酵對金黃色葡萄球菌的抑制能力評估 67
5.3.3牛樟芝合併發酵對青枯菌的抑制能力評估 79
5.3.4 HPLC 分析 90
第六章 結論 93
第七章 參考文獻 94

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