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研究生:盧立章
研究生(外文):Lu, Li-Chang
論文名稱:不同季節與地區產台灣蜂膠之抗菌活性及其DPPH自由基清除能力
論文名稱(外文):The Antibacterial and DPPH Radicals Scavenging Effects of Taiwanese Propolis Obtained from Different Regions and Seasons
指導教授:周正俊周正俊引用關係陳裕文陳裕文引用關係
指導教授(外文):Cheng-Chun Chou, Ph. D.Yue-Wen Chen, Ph. D.
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:食品科技研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:73
中文關鍵詞:蜂膠地區季節抗菌活性DPPH自由基
外文關鍵詞:propolisregionsseasonsantibacterial activitiesDPPH radiacals
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蜂膠是蜜蜂蒐集特定植物樹皮或葉芽所分泌的樹脂,混入蜜蜂分泌之酵素,並結合蜂蠟及花粉所製成的膠狀物質。本研究探討台灣不同地區(北-台大,中-南投名間鄉,南-屏東枋寮鄉)、不同季節(六月、八月、十~十一月)所收集蜂膠之抗菌活性及其清除DPPH自由基之能力。
結果發現,蜂膠之香膠(balsam, 80%酒精萃出物)含量、抗菌效果及a-diphenyl-b-picryl-hydrazyl (DPPH)自由基的清除能力因產地、產季之不同而有差異。大致而言,無論其產地,香膠含量以六月產者最高,十~十一月產者最低。此外名間產者其香膠含量最高。
不同病原菌對蜂膠的敏感性也不同,Staphylococcus aureus及Listeria monocytogenes對蜂膠較為敏感,而對Salmonella typhimurium及Escherichia coli O157:H7則否。大致而言,以六月產台灣蜂膠之抗菌效果較佳。在所有蜂膠樣品中則以名間六月產蜂膠之抗菌活性最強。
進一步對Sta. aureus測試顯示,蜂膠酒精萃出物在37℃時所表現之抗菌效果比在低溫時(4、25℃)佳。且其抗菌效果在酸性環境下(pH 5.0、6.0)亦比在高pH環境下(pH 8.0、9.0)強。菌齡影響Sta. aureus對蜂膠酒精萃出物之敏感性,其敏感性依序為對數期中期、末期及定常期之菌體。此外,蜂膠酒精萃出物會造成Sta. aureus胞內260 nm吸光物質(核酸、DNA、RNA)的洩漏。蜂膠中的抗菌物質具有良好的熱安定性。蜂膠亦具DPPH自由基清除之能力,所測試之台灣產蜂膠中以六月產者具有較佳之DPPH自由基清除能力。
Propolis is a resinous substance collected by honeybees from buds and leaves of trees and plants, mixing with pollen, bee wax as well as enzymes secreted by bees. In the present study, the antibacterial activities and DPPH (a-diphenyl-b-picryl-hydrazyl) radicals scavenging effects of Taiwanese propolis obtained from various regions (NTU, Mingchien and Fangliao) at different seasons (June, August, and October-November), was examined.
Results showed that the balsam content, antibacterial activities and DPPH radicals scavenging effects of the propolis were varied with the harvesting regions and seasons. In general, the balsam content of propolis obtained in June was the highest, while those collected in October-November was the lowest, regardless of harvesting regions. The balsam content of propolis obtained from Mingchien in June was the highest.
The pathogens tested showed various extents of susceptibility to ethanolic extract of propolis (EEP). Staphylococcus aureus and Listeria monocytogenes were found susceptible while Salmonella typhimurium and Escherichia coli were resistant to propolis. The Taiwanese propolis collected in June possessed a higher antibacterial activity than those collected in other seasons, and the Mingchien propolis collected in June exhibited the highest antibacterial activity among all of Taiwanese propolis tested.
Further test on Sta. aureus revealed that EEP showed stronger antibacterial activity at 37℃ than at 4℃or 25℃, in acid environment (pH 5.0 and pH 6.0) than in higher pH environment (pH 8 and pH 9). Cell age also affected the susceptibility of Sta. aureus to EEP. Cells in the mid-exponential phase were most sensitive, followed by the late exponential phase cells and stationary phase cells. In addition, it was also noted that EEP caused the leakage of 260-nm-absorbing materials (nucleotide, DNA, RNA) from Sta. aureus cells and possessed good thermal stability. Furthermore, EEP showed scavenging effect on DPPH radicals. Among the propolis tested, the Taiwanese propolis collected in June exhibited the highest DPPH scavenging activity.
摘要-------------------------------------------------------I
Abstract--------------------------------------------------II
目錄------------------------------------------------------IV
圖目錄------------------- --------------------------------VII
表目錄---------------------------------------------------VIII
壹、前言---------------------------------------------------1
貳、文獻整理-----------------------------------------------3
一、蜂膠的來源---------------------------------------------3
(1)膠源植物------------------------------------------------3
(2)產膠蜂種------------------------------------------------4
(3)蜂膠的取得----------------------------------------------4
(4)蜂膠之萃取方法------------------------------------------6
二、蜂膠之物理化學性質-------------------------------------6
(1)蜂膠之物理性質------------------------------------------6
(2)蜂膠之化學組成------------------------------------------7
A.蜂膠之組成份---------------------------------------------7
B.蜂膠中之類黃酮-------------------------------------------7
三、蜂膠抗微生物之活性-------------------------------------9
(1)蜂膠之抗細菌活性----------------------------------------9
(2)影響蜂膠抗菌效用之因素---------------------------------11
A.產地----------------------------------------------------11
B.產季----------------------------------------------------12
C.菌株----------------------------------------------------12
(3)蜂膠之抗真菌活性---------------------------------------13
(4)蜂膠之抗濾過性病毒活性---------------------------------14
四、蜂膠抑菌作用之機制------------------------------------14
(1)蜂膠中之抗菌成分---------------------------------------14
(2)蜂膠對菌體的作用---------------------------------------15
五、蜂膠之抗氧化能力--------------------------------------16
六、蜂膠之其他生物活性與醫療效用--------------------------17
(1)蜂膠的抗腫瘤活性---------------------------------------17
(2)蜂膠之抗發炎作用---------------------------------------17
(3)蜂膠之其他活性-----------------------------------------18
七、蜂膠在使用上之安全性----------------------------------19
(1)毒性評估-----------------------------------------------19
(2)蜂膠中的重金屬-----------------------------------------19
(3)過敏反應-----------------------------------------------20
八、蜂膠在商業上之應用------------------------------------20
九、台灣蜂膠的前瞻性--------------------------------------20
參、材料方法----------------------------------------------22
一、實驗材料----------------------------------------------22
(1)菌種---------------------------------------------------22
(2)蜂膠---------------------------------------------------22
(3)培養基-------------------------------------------------22
(4)化學藥品-----------------------------------------------22
(5)儀器及器材---------------------------------------------23
二、實驗方法----------------------------------------------23
(1)菌株之保存與更新---------------------------------------23
(2)菌株之活化與接種源之製備-------------------------------24
(3)蜂膠酒精萃出液之製備-----------------------------------24
(4)香膠(balsam)含量分析-----------------------------------24
(5)蜂膠酒精萃出物對不同病原菌的抗菌試驗-------------------25
(6)蜂膠酒精萃出物對金黃色葡萄球菌的抗菌作用---------------25
(7)溫度對蜂膠酒精萃出物於金黃色葡萄球菌抗菌作用之影響-----26
(8)蜂膠酒精萃出液對不同菌齡之金黃色葡萄球菌的抗菌作用-----26
(9)不同pH對蜂膠酒精萃出物於金黃色葡萄球菌抗菌作用之影響---27
(10)溫度處理對蜂膠酒精萃出物抑制金黃色葡萄球菌
生長效應之影響----------------------------------------27
(11)蜂膠酒精萃出物對金黃色葡萄球菌菌體內
260nm吸光物質洩漏之影響-------------------------------28
(12)菌數測定----------------------------------------------28
(13)蜂膠酒精萃取液對DPPH自由基清除效應--------------------28
(14)統計分析----------------------------------------------29
肆、結果與討論--------------------------------------------30
一、不同來源蜂膠及其萃取液感官特性之差異------------------30
二、不同季節產地蜂膠之酒精萃取率--------------------------33
三、蜂膠酒精萃出物對病原菌之抑菌效果----------------------35
(1)不同病原菌對蜂膠酒精萃出物之敏感性---------------------35
(2)蜂膠酒精萃出物對金黃色葡萄球菌之抑菌效果---------------38
A.不同產地、季節生產之蜂膠及其劑量對金黃色葡萄球菌
之抗菌效果----------------------------------------------38
B.蜂膠對金黃色葡萄球菌之最低抑菌濃對及最低殺菌濃度--------45
四、蜂膠酒精萃出物在不同條件下對金黃色葡萄球菌之抑菌效果--47
(1)培養溫度-----------------------------------------------47
(2)菌齡---------------------------------------------------49
(3)pH-----------------------------------------------------51
五、金黃色葡萄球菌經蜂膠酒精萃出物作用後其細胞內
260 nm吸光物質之洩漏情形------------------------------53
六、溫度處理對蜂膠酒精萃出物抑菌效果之影響----------------53
七、不同產地及產季蜂膠酒精萃出物對DPPH 自由基之清除能-----55
伍、結論--------------------------------------------------60
陸、參考文獻----------------------------------------------61
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