臺灣博碩士論文加值系統

以細菌性纖維及海藻酸鈉重組之濕態重組生物薄膜(wet fabricated bio-film, WFBF)水分含量高，於室溫下長期儲存易有細菌及黴菌滋生而變質。由於使用單一抗菌物質所需劑量較高，且抑菌種類有限，因此本研究使用天然抗菌劑幾丁聚醣(chitosan, Chi)及鏈黴菌素(natamycin, NAT)，探討此兩種抗菌及抗黴劑複合對黑麴菌(Aspergillus niger, BCRC 30506)、金黃色葡萄球菌(Staphylococcus aureus, BCRC 10780) 、大腸桿菌(Escheriachia coli, BCRC 10675)及綠膿桿菌(Pseudomona aeruginosa, BCRC 11633)的抗黴、抗菌能力，並模擬市售濕態面膜儲存方式，將WFBF浸漬於複合抗菌液中，探討是否達到協同抗菌效應。由微孔盤及抑菌環實驗結果顯示，NAT對A. niger的最低抑制濃度(minimum inhibitory concentration, MIC)為375 µg/mL，但對S. aureus、E. coli及P.aeruginosa無抑菌效果；Chi對S. aureus、E. coli及P. aeruginosa MIC分別為3,200 µg/mL、6.25 µg/mL及100 µg/mL，但對A. niger無抑黴效果。將Chi與NAT複合添加200/188 µg/mL即對A. niger有抑菌效果，可降低兩種抗菌物質劑量且具有協同作用。為考慮抑制S. aureus，於薄膜儲存試驗中，將Chi劑量提升，發現於Chi與NAT複合濃度1,600/47 µg/mL對S. aureus及E. coli抑菌能力達168小時，且在同樣複合條件下，可抑制A. niger至少21天不生長且皆具有協同效果，然而對P. aeruginosa在Chi與NAT複合濃度1,600/188 µg/mL下，雖具抑菌能力但效果較不明顯。在物理性質方面，複合抗菌液對浸漬24小時的WFBF機械性質並無影響，水蒸氣通透速率下降，但長時間儲存後的質地有軟化趨勢。以此複合抗菌液儲存WFBF可降低兩種抗菌物質的使用劑量，具有取代化學防腐劑的潛力。

The bacteria and mold can be observed on wet fabricated bio-film (WFBF) contaning high moisture content during long-term storage at room temperature. But treatment with single antibacterial substance with narrow targets usually requires high-dose and can be a problem. Therefore, this study investigated the combinational antibacterial effect of natural antibacterial and antifungal substances, chitosan (Chi) and natamycin (NAT), against Aspergillus niger, Staphylococcus aureus, Escheriachia coli and Pseudomona aeruginosa. In addition, the WFBF made of bacterial cellulose (BC) was immersed in combinational antibacterial solution to simulate the storage conditions of wet masks in market, and the synergistic effect was investigated. According to the 96-well microplate and inhibition zone tests, the minimum inhibitory concentration (MIC) of NAT against A. niger was 375 µg/mL, but ineffective against S. aureus, E. coli and P. aeruginosa. The MIC of Chi against S. aureus, E. coli and P. aeruginosa were 3,200 µg/mL, 6.25 µg/mL and 100 µg/mL, respectively, but ineffective against A. niger. The Chi/NAT under 200/188 µg/mL combinational ratio exhibited effective inhibition against A. niger, which reduced the dosage of antimicrobial substances and showed synergistic effect. For inhibition of S. aureus, high-dose of Chi was required during storage. The Chi/NAT dosage at 1,600/47 µg/mL inhibited the growth of S. aureus and E. coli within 168 hrs. Moreover, this dosage of Chi/NAT effectively inhibited the growth of A. niger for at less 21 days and exhibited synergistic effect. However, such dosage of Chi/NAT (1,600/188 µg/mL) could not effectively inhibit the growth of P. aeruginosa. In physical properties, there was no difference in the mechanical properties of WFBF within 24 hr storage, but water vapor transmission rate decreased. The WFBF was softened as prolonged the storage. These results indicated the combinational antibacterial effect of Chi and NAT can lower the dosage of antibacterial substances, and possesses the potential for substituting chemical preservatives to extend shelf life.

中文摘要 I
Abstract II
謝誌 IV
目錄 V
表目錄 VIII
圖目錄 IX
附表目錄 XI
附圖目錄 XII
一、前言 1
二、文獻回顧 2
1. 細菌性纖維素(bacterial cellulose, BC) 2
1.1. BC的特性及其結構 2
1.2. BC來源及其生合成 2
1.3. BC複合材料之應用 3
1.4. 面膜及其保存 3
2. 天然抗菌物質 4
2.1. 鏈黴菌素(natamycin, NAT) 4
2.1.1. NAT的特性及其結構 4
2.1.2. NAT抑菌機制 4
2.1.3. NAT應用 5
2.2. 幾丁聚醣(chitosan, Chi) 5
2.2.1. Chi的結構及其特性 5
2.2.2. Chi抑菌機制 6
2.2.3.Chi應用 6
3. 複合抗菌 7
3.1. NAT複合其他抗菌物質之應用 7
3.2. Chi 複合其他抗菌物質之應用 7
3.3. NAT與Chi複合抗菌之應用 8
三、材料與方法 9
1. 實驗架構 9
2. 實驗材料與方法 10
2.1. 實驗菌株及培養方法 10
2.2. 實驗菌種保存 10
2.3. 天然抗菌物質製備 10
2.3.1. NAT製備 10
2.3.2. Chi製備 10
2.4. 最低抑菌濃度(minimum inhibitory concentration, MIC)測定 11
2.4.1. NAT MIC測定 11
2.4.2. Chi MIC測定 11
2.4.3. Chi及NAT複合抑黴試驗 11
2.4.4. 複合抑黴及抑菌效果評估 11
2.5. 濕態重組薄膜之製備 12
2.5.1. BC 前處理 12
2.5.2. 重組薄膜製備 12
2.6. 薄膜儲存試驗 12
2.7. 物理性質分析 13
2.7.1. 拉伸強度(tensile strength)與伸張率(elongation at break) 13
2.7.2. 水蒸氣穿透速率測定(Water Vapor Transmission Rate, WVTR) 13
2.7.3. 掃描電子顯微鏡(scanning electron microscope, SEM) 14
2.8. 統計分析 14
四、結果與討論 15
1. 選擇NAT及Chi適當之抑菌條件 15
1.1. NAT對A. niger、S. aureus、E. coli及P. aeruginosa抑黴及抑菌效果 15
1.2. Chi對A. niger之抑黴效果 15
1.3. Chi對S. aureus、E. coli及P. aeruginosa之抑菌效果 16
2. Chi及NAT對A. niger複合抑黴 16
3. 複合抑菌、抑黴之儲存試驗 17
3.1. Chi/NAT複合對S. aureus、E. coli及P. aeruginosa的影響 17
3.2. Chi/NAT複合對A. niger之抑黴效果 18
4. WFBF浸漬於不同抗菌溶液之物理性質分析 18
5. 細菌對浸漬於Chi/NAT複合抗菌液之WFBF細微結構變化 19
五、結論 20
六、參考文獻 21
七、表 34
八、圖 36
九、附表 57
十、附圖 58

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