各試驗乳酸菌株對Caco-2 cell 之侵入百分比如表 4.7 所示，由結 果可知本研究中所試驗之乳酸菌株均對Caco-2 cell 不具侵入性。另
外，將試驗菌株以pH 4 酸液及 0.1%膽鹽溶液連續作用後，各試驗菌
株對Caco-2 cell 之侵入百分比如表 4.8 所示，由結果發現將乳酸菌以 模擬胃腸道條件作用後之乳酸菌仍對Caco-2 cell 不具侵入性，因此可 進一步推論本試驗所使用之菌株對人體具安全性，不具侵襲人體腸道 細胞之危險性。
行政院衛生署公告可做為食品原料或食品加工使用之微生物包 括本研究中所使用的三種乳酸菌－L. acidophilus、B. bifidum、L.
paracasei (# 2.)。具侵襲力的致病菌能突破宿主黏膜屏障，侵入體內
定殖造成腸道疾病，因此成為益生菌之條件必須具有吸附人類腸道細 胞之能力，但不能具有腸道細胞侵入性，若菌株具侵入性可能同時擁 有相當程度之病原性與感染性(Ford et al., 1996; Urao et al., 1996)。
表 4.7、各試驗乳酸菌株侵入 Caco-2 cells 之百分比 Table 4.7. Invasion rate of the tested LAB
*N.D.: not detected.
Strain Invasion rate (%)
L. acidophilus N.D.*
B. bifidum N.D.
L. paracasei N.D.
表4.8、各試驗乳酸菌株經 pH 4 酸液作用 3 hr 後再以 0.1% 膽鹽 連續處理 3 hr 後侵入 Caco-2 cells 之百分比
Table 4.8. Invasion rate of the tested LAB after being treated with pH 4 acid solution for 3 hr and then 0.1% bile for 3 hr continually
*N.D.: not detected.
Strain Invasion rate (%)
L. acidophilus N.D.*
B. bifidum N.D.
L. paracasei N.D.
第五章 結論
本研究利用 L. acidophilus BCRC 10695, B. bifidum BCRC 14615, L. paracasei BCRC 14023 作為試驗菌株，探討益生菌經酸及膽鹽作用後
其耐受性、模擬腸道吸附性、拮抗 C. perfringens BCRC 13019 之能力及 菌株安全性等基礎益生性質，及菌株受模擬腸胃道不良環境因子條件作 用後對其細胞膜完整性之影響，總結試驗結果如下：
(1)所有試驗乳酸菌株於酸及膽鹽之個別試驗中，L. acidophilus 表現最佳 耐酸性，L. paracasei 表現最佳膽鹽耐受性；而結合酸與膽鹽之連續
性試驗結果，所有試驗菌株於pH 4 酸液及 0.1%膽鹽之連續作用下，
具有最高之存活率。
(2)未經酸及膽鹽作用之 L. acidophilus 擁有最佳吸附能力(8.28%)；經不
同濃度之酸及膽鹽溶液作用後，所有試驗菌株僅於pH 4 酸液及 0.1%
膽鹽條件作用後存活之乳酸菌仍保有吸附能力，且其吸附能力大於未 經酸及膽鹽作用者；其中又以 L. paracasei 擁有最佳之吸附能力 (80.68%)。
(3)本研究結果推論，低 pH 值與高濃度膽鹽會導致乳酸菌株存活率下 降，且其造成之傷害會使菌株喪失其吸附能力。
(4)預防 C. perfringens 侵襲腸道之吸附抑制試驗，不論 C. perfringens 是
否預先以pH 4 酸液及 0.1%膽鹽連續作用，試驗乳酸菌株皆可有效預
防 C. perfringens 侵襲腸道細胞，且乳酸菌對預先以酸及膽鹽處理之 病原菌具有較佳但未達顯著性之拮抗能力。
(5)取代 C. perfringens 吸附於腸道細胞之試驗結果顯示，不論乳酸菌是
否預先以pH 4 酸液及 0.1%膽鹽連續作用，試驗菌株皆可有效降低預
先吸附於Caco-2 cell 上 C. perfringens 之吸附率，因此達到取代 C.
perfringens 吸附於腸道細胞之效果；但經模擬胃腸道條件作用後之乳
酸菌其拮抗能力略低於未經作用之乳酸菌。
(6) β-半乳糖苷酶活性試驗結果得知，隨著膽鹽濃度之提高 β-半乳糖苷 酶活性也增加，顯示菌體細胞膜可能遭受之破壞程度隨膽鹽濃度提高 而增加。
(7)生體外試驗得知不論是否經酸與膽鹽作用，L. acidophilus、B.
bifidum、L. paracasei 皆不具細胞侵入性，因此初步推論其對人體腸
道不具侵襲性。
(8)本研究結果顯示，以酸及膽鹽作為模擬胃腸道之環境條件用以探討乳 酸菌經上述條件作用後之益生性質表現，應為一適當可行之生體外試 驗評估模式。
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