臺灣博碩士論文加值系統

自八十七年七月至八十八年一月間，就北市某速食連鎖店進行生菜沙拉微生物品質分析。在抽檢63件生菜沙拉檢體中，生菌數以103-104 CFU/g者最多，共計22件；檢體中有30件檢出大腸桿菌群，而金黃色葡萄球菌及大腸桿菌在所有的生菜沙拉檢體中均未被檢出。
Lactococcus lactis subsp. cremoris、Lactococcus lactis subsp. lactis及Pediococcus pentosaceu所產的三種粗細菌素液經過減壓濃縮十倍後，對Aeromonas hydrophila、Escherichia coli O157:H7 、Listeria monocytogenes 、Staphylococcus aureus 及Salmonella enteritidis均有良好的抑制效果（p＜0.05）。於7℃時，粗細菌素液抑制食品病原菌的效果優於25℃。粗細菌素濃縮液於pH 3的環境下，抑制病原菌的效果最佳，越趨近中性環境，抑菌活性均顯著下降。在相同pH環境時，P. pentosaceus的十倍濃縮粗細菌素液的抑菌效果最佳，對於pH值變化的耐受力較強，即使在pH 7時，仍具抑制病原菌的活性。
以將含病原菌的濾紙浸泡P. pentosaceus的十倍濃縮粗細菌素液的方式抑制病原菌效果最明顯，浸泡60分鐘可將A. hydrophila、E. coli O157:H7、L. monocytogenes與S. aureus完全殺滅；浸泡120分鐘能降低S. enteritidis2.6個對數值。將P. pentosaceus的十倍濃縮粗細菌素液噴灑含病原菌的濾紙亦有部分抑制病原菌的效果，噴灑120分鐘後將A. hydrophila、E. coli O157:H7與S. aureus的菌數降至無法檢出，降低L. monocytogenes約2.6個對數值。而未經濃縮的P. pentosaceus的粗細菌素液抑制病原菌的效果最差，即使120分鐘後，對於A. hydrophila、L. monocytogenes與S. aureus幾乎沒有抑制能力，僅對E. coli O157:H7與S. enteritidis有顯著的抑制能力。

During July 1998 through January 1999, vegetable salads had been sampled from fast-foods chain stores in Taipei for analysis. Results indicated that Coliforms was detected in 30 samples, and the total aerobic count of 103-104 CFU/g were found in 22 out of 63 samples. E. coli and S. aureus were not found in all samples.
Tenfold concentrated crude bacteriocin solutions produced from Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis and Pediococcus pentosaceus inhibited Aeromonas hydrophila, Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus and Salmonella enteritidis (p<0.05). The inhibition effect of at 7℃was much better than that at 25℃. The antimicrobial activity of concentrated bacteriocin solutions was best at pH 3, however it decreased with increasing pH. Tenfold concentrated bacteriocin solution produced by P. pentosaceus had the strongest inhibition effect. It was stable at various pH values and still possessed the ability to inhibit pathogens even at pH7.
Immersion of filter paper containing pathogens in tenfold concentrated crude bacteriocin solutions produced by P. pentosaceus effectively inhibited the growth of pathogens. After immersion for 60 min, it completely destroyed A. hydrophila, E. coli O157:H7, L. monocytogenes and S. aureus negative and reduced 2.6 log CFU/g of S. enteritidis. Spraying filter paper containing pathogens with tenfold concentrated bacteriocin solutions produced by P. pentosaceus showed complete inhibition of A. hydrophila, E. coli O157:H7 and S. aureus after 120 min of treatment. It also reduced 2.6 log CFU/g of L. monocytogenes. Crude bacteriocin solution without concentration significantly inhibited E. coli O157:H7 and S. enteritidis, it did not inhibit the growth of A. hydrophila, L. monocytogenes and S. aureus, even after 120 min of treatment.

第一章、緒言……………………………………………………………1
第二章、文獻回顧………………………………………………………3
一、輕度加工蔬果…………………………………………………3
二、輕度加工的定義………………………………………………4
三、食品中病原菌污染輕度加工蔬菜之情形……………………4
四、輕度加工食品中常見微生物或病原菌之簡介………………21
(一)氣單胞菌屬（Aeromonas）…………………………….…21
(二)大腸桿菌屬（Coliforms）...………………………………22
(三)大腸桿菌（E. coli）……………………………..………23
(四)E. coliO157 : H7…………………………………………23
(五)單核細胞增生李斯特氏菌（L. monocytogenes）..………27
(六)沙門氏桿菌屬（Salmonella）………………………..….29
(七) 金黃色葡萄球菌（S. aureus）…………………………30
五、抑制輕度加工蔬果中微生物的方法.…………..……………31
(1)物理方法.…………………………………………………32
Œ高強度脈衝電場………………………………………33
振盪磁場………………………………………………33
Ž高強度脈衝光線………………………………………34
超音波…………………………………………………34
高靜水壓………………………………………………35
(2)化學方法.…………………………………………………35
(3)生物控制法.………………………………………………39
五、乳酸菌的簡介………………………………………………..40
六、乳酸菌抑菌機制……………………………………………..43
(1)有機酸的產生與pH值的降低………………….……….43
(2)二氧化碳的作用………………………………………….44
(3)過氧化氫的累積………………………………………….44
(4)雙乙醯…………………………………………………….46
(5)Reuterin…………………………………………………...46
(6)細菌素…………………………………………………….46
七、細菌素的介紹……………….……………………………….47
(1)細菌素的發現………………………………….…………47
(2)影響細菌素生產的因子……………………….…………49
(3)細菌素抑制病原菌之機制…………………….…………52
第三章、材料與方法…………………………………….…………….54
一、實驗材料…………………………………………….……….54
(1)市售生菜沙拉…………………………………….………54
(2)菌種…………………………………………………….…54
(3)培養基…………………………………….………………54
(4)化學試藥與材料……………………………….…………55
二、儀器設備………………………………………….………….55
三、實驗方法及步驟……………………………………….…….56
(1)稀釋緩衝溶液的製備…………………….………………56
(2)菌種保存、更新與接種源之製備……………….………56
(3)粗細菌素液的製備………………………….……………57
(4)粗細菌素十倍濃縮液的製備………………...…………..57
(5)抑菌活性測試……………………..………………….…..57
(6)速食連鎖店生菜沙拉微生物品質調查……….…………58
1.採樣方法…………………………………………….…..58
2.生菌數之測定………………………………...…..……..58
3.Coliforms之檢測…………………….………….………58
4. E. coli的測定…………………………….……………..59
5. S. aureus之檢測…………………………………....…..59
(7)產細菌素乳酸菌抑制病原菌試驗……...…………..……60
1.粗細菌素溶液濃度與抑菌效果試驗……………….…60
2.反應溫度對粗細菌素溶液與抑菌效果試驗……….…60
3.粗細菌素液之pH值的抑菌效果試驗…………..…….61
(8)模式系統下粗細菌素液抑制病原菌試驗……………….61
1.處理方式與反應時間對P. pentosaceus的粗細菌素
溶液抑制病原菌的影響………………..…….….……61
2.微生物之檢測………..………………………………...62
四、統計分析…………………………….……………………….62
第四章、結果與討論………………………….……………………….63
一、速食連鎖店生菜沙拉微生物品質調查……………….…….63
二、粗細菌素液抑制病原菌之比較………….………………….69
（一）粗細菌素溶液濃度……………………………………69
（二）反應溫度……………………………………………….73
（三）反應pH值…………………………………….……….78
三、以濾紙模式系統探討粗細菌素液抑制病原菌試驗….……..92
第五章、結論……………………………………………………….…104
參考文獻…………………………………………………………...….106
圖 目 錄
頁次
圖一、截切蔬果與其他食品保藏方式之比較……………………….5
圖二、輕度加工蔬菜之製造流程………………………………….….6
圖三、乳酸發酵………………………………………………….…….42
圖四、乳酸菌產生過氧化氫之反應機制……………………….…….45
圖五、乳酸鏈球菌素的形成及其分子結構…………….……………..50
圖六、P. pentosaceus的十倍濃縮粗細菌素液抑制S. aureus情形…72
圖七、十倍濃縮粗細菌素液於不同pH值對A. hydrophila抑菌效果之影響….…………………………………………………………79
圖八、十倍濃縮粗細菌素液於不同pH值對E. coli O157:H7抑菌效果
之影響……………………………………………………….....82
圖九、十倍濃縮粗細菌素液於不同pH值對L. monocytogenes 抑菌效果之影響………………………….……………………………85
圖十、十倍濃縮粗細菌素液於不同pH值對S. aureus抑菌效果之影響……………………………………………….………………88
圖十一、十倍濃縮粗細菌素液於不同pH值對S. enteritidis抑菌效果之影響………………………………………….……..………90
圖十二、P. pentosaceus的粗細菌素液的處理方式與反應時間抑制A. hydrophila的情形…………………………………………….94
圖十三、P. pentosaceus的粗細菌素液的處理方式與反應時間抑制E. coli O157:H7的情形……………….………………………...96
圖十四、P. pentosaceus的粗細菌素液的處理方式與反應時間抑制L. monocytogenes的情形………………………………………..98
圖十五、P. pentosaceus的粗細菌素液的處理方式與反應時間抑制S. aureus的情形………………………………………………...99
圖十六、P. pentosaceus的粗細菌素液的處理方式與反應時間抑制S. enteritidis的情形……………………………………………102
表 目 錄
頁次
表一、常見蔬菜組成份…………………………………………………..8
表二、蔬菜經截切後微生物含量變化情形………………………..……9
表三、製備沙拉之市售萵苣的生菌數分布情形………………………11
表四、生鮮蔬菜及沙拉中沙門桿菌、志賀氏桿菌及生菌數之
檢出情形……………………………………………………….12
表五、生鮮蔬菜中致病菌之污染情形…………………………………13
表六、大阪超市蔬菜沙拉之微生物數…………………………...….…14
表七、氣單胞菌污染市售葉菜類蔬菜之情形…………………………16
表八、不同採購地點之沙拉常用生菜中李斯特菌檢出情形…………17
表九、中西式速食中微生物檢出情形之比較……………………...…18
表十、台灣不同地區生菜沙拉之金黃色葡萄球菌檢出情形…….…..19
表十一、生菜沙拉中大腸菌群及大腸桿菌之檢出情形…………..…..20
表十二、腸毒性大腸桿菌的主要特性………………………………...25
表十三、溫度對培養於trypticase soy broth中E.coli O157：H7生長
之影響………………………………………………………..26
表十四、截切蔬菜與甜瓜之過氧化氫殘留量………………………...38
表十五、各種細菌素名稱及其生產菌株………………………………51
表十六、生菜沙拉中金黃色葡萄球菌、大腸桿菌群及大腸桿菌之檢
出情形………………………………………………………...65
表十七、生菜沙拉中大腸桿菌群之檢出情形…………………………66
表十八、生菜沙拉中生菌數之檢出情形………………………………67
表十九、生菜沙拉種類中大腸桿菌群之檢出情形……………………68
表二十、粗細菌素與其十倍濃縮溶液抑制食品病原菌之效果………70
表二十一、溫度對L. lactis subsp. lactis 的十倍濃縮粗細菌素液抑
制病原菌之影響…………………………………………..74
表二十二、溫度對P. pentosaceus的十倍濃縮粗細菌素液抑制病原菌
之影響……………………………………………………..76
表二十三、溫度對L. lactis subsp. cremoris的十倍濃縮粗細菌素液抑
制病原菌之影響…………………………………………..77

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