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中國畜牧學會會誌
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| 篇名 | 芽孢桿菌之抗病毒活性 |
|---|---|
| 卷期 | 46增刊 |
| 並列篇名 | Antiviral ability of Bacillus strains |
| 作者 | 陳立維 、 劉嚞睿 |
| 頁次 | 232-232 |
| 關鍵字 | 抗病毒 、 芽孢桿菌 、 辛德比病毒 、 Antiviral activity 、 Bacillus 、 Sindbis virus |
| 出刊日期 | 201712 |
中文摘要
益生菌為在生物體內達一定數量後可以對宿主造成正面影響的微生物，常見之益生菌以乳酸菌與芽 孢桿菌為主。由於芽孢桿菌在高溫下較易存活，且在嚴苛環境下能形成內孢子，因此在製作產品過 程中菌體存活數量往往較乳酸菌為佳。本次研究將探討芽孢桿菌的抗病毒能力，所使用的三株桿菌 分別為枯草桿菌AC（Bacillus subtilis AC）、地衣桿菌CK（B. licheniformis CK）以及解澱粉芽孢 桿菌 LN（B. amyloliquefaciens LN）。本研究使用的病毒為經基因改造之辛德比病毒，細胞受該病 毒感染後會表現綠色螢光蛋白。根據先前的研究顯示，比較辛德比病毒對多種細胞株之感染能力， 以倉鼠腎臟細胞為最易受感染，故以倉鼠腎臟細胞為本研究之模型。實驗將芽孢桿菌分為四個部 分，分別為細胞壁、胞內萃取物、胞外分泌物和活菌，在分別與細胞進行共培養 24 小時後，分別 以四甲基偶氮唑鹽（3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT）及 7 氨基放 線菌素 D（7-aminoactinomycin D, 7AAD）檢測細胞毒性。確認芽孢桿菌不具細胞毒性時，則進一 步將病毒加入細胞進行感染 24 小時，再以流式細胞儀檢測受感染的細胞比例。結果顯示解澱粉芽 孢桿菌 LN 的所有處理均具有細胞毒性，而地衣桿菌CK 活菌以及枯草桿菌 AC 胞外分泌物亦 具有細胞毒性，而 CK 胞內萃取物不具有細胞毒性，且具有顯著的抗病毒活性，其機制需進一步 研究闡明。此結果顯示地衣桿菌 CK 的胞內物質在此倉鼠腎臟細胞模型下具有抵抗辛德比病毒的 潛力。
英文摘要
Probiotics are live microbes, which when administered in adequate amounts confer a health benefit to the host. The most commonly used probiotics include Lactobacillus and Bacillus. Because of spores being heat-stable, bacilli have advantages over other non-spore formers such as Lactobacillus spp. as the product can be stored at room temperature in a desiccated form without any loss. The aim of this study is to determine whether the Bacillus strains including Bacillus subtilis AC, B. licheniformis CK, and B. amyloliquefaciens LN, have antiviral ability against Sindbis virus (SBV). In a previous study, the susceptibilities of several cell model to the infection of the recombinant SBV which expressed enhanced green fluorescence protein (EGFP) were investigated, and the results indicated that baby hamster kidney (BHK) cells were most susceptible to SBV infection. Therefore, BHK were used as the cell model in this study. Four Bacillus cell fractions, including cell wall, intercellular extract, extracellular extract, and whole viable bacterial cell, were co-cultured with BHK cell for 24 hours and then the cytotoxicity were determined by the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and 7AAD (7-aminoactinomycin D) assays. Antiviral activity of each Bacillus cell fractions were determined after challenge the BHK cells with virus for 24 hours by flow cytometry. We found that the all fractions of LN have cytotoxicity. The extracellular extract of AC and viable CK also have cytotoxicity. In the antiviral activity test, only the intercellular extract of CK was able to significantly reduce the rate of viral infection. Further study will be conducted to investigate the antiviral mechanisms of CK. These results demonstrated that the intercellular extract of B. licheniformis CK has a potential to be used to protect viral infection.