Use of a spectrophotometric bioassay for determination of microbial sensitivity to manuka honey
Use of a spectrophotometric bioassay for determination of microbial sensitivity to manuka honey
复制标题
DOI：
10.1016/j.mimet.2005.04.007
复制
发表时间：
2006-01-01
影响因子：
2.2
通讯作者：
Moran, N
中科院分区：
文献类型：
--
作者：
Patton, T;Barrett, J;Moran, N
麦卢卡蜂蜜的抗微生物活性已被充分记载（Molan，1992 a，B，c，1997）[Molan，P.C.，1992.蜂蜜的抗菌活性1：抗菌活性的性质。蜜蜂世界73（1）5-28;莫兰，P.C.，1992.蜂蜜的抗菌活性2：抗菌活性效力的变化。蜜蜂世界73（2）59-76; Molan，P.C.，1992.蜂蜜的药用价值养蜂人季刊26;莫兰，P.C.，1997.寻找具有出色抗菌和抗真菌活性的新西兰蜂蜜。新西兰养蜂人4（10）20-26]。目前用于测定这种抗微生物效果的生物测定采用良好的扩散（Ahn和Stiles，1990）[Ahn，C.，斯泰尔斯法医1990.真空包装肉制品中乳酸菌的抗菌活性研究。Journal of Applied Bacteriology 69，302-310]，（韦斯顿等人，1999）[韦斯顿，R. J.，米切尔，K.R.，艾伦，K.L.，1999.新西兰麦卢卡蜂蜜的抗菌酚类成分。J. Food Chem.64，295-301]或圆盘扩散（陶尔米纳等人，2001）[陶尔米纳，彼得J.，Niemira，Brendan A.，Beuchat，Larry R.，2001.蜂蜜对食物家庭病原体的抑制活性受过氧化氢和抗氧化能力水平的影响。国际食品微生物学杂志69，217-225]使用抑制区作为细菌敏感性的指示剂的测定。采用96孔微量滴定板的24小时分光光度测定法的开发，比目前采用的测定法更灵敏，更适合统计分析。这种简单而快速的测定允许广泛的动力学研究，即使在低蜂蜜浓度的存在下，并且能够检测低于孔或盘扩散测定记录的抑制水平。在本文中，我们比较了测定两个孔和盘扩散测定。我们获得的结果为分光光度法MIC值表明，该方法具有更高的灵敏度比标准的孔和盘扩散测定。此外，还研究了该方法的试验间和试验内方差，证明了该方法的重现性和重复性。(c)2005 Elsevier B. V.保留所有权利。
The antimicrobial activity of manuka honey has been well documented (Molan, 1992a,b,c, 1997) [Molan, P.C., 1992. The antibacterial activity of honey. 1: the nature of the antibacterial activity. Bee World 73 (1) 5-28; Molan, P.C., 1992. The antibacterial activity of honey. 2: variation in the potency of the antibacterial activity. Bee World 73 (2) 59-76; Molan, P.C., 1992. Medicinal uses for honey. Beekeepers Quarterly 26; Molan, P.C., 1997. Finding New Zealand honeys with outstanding antibacterial and antifungal activity. New Zealand Beekeeper 4 (10) 20-26]. The current bioassays for determining this antimicrobial effect employ a well diffusion (Ahn and Stiles, 1990) [Ahn, C., Stiles, M.E., 1990. Antibacterial activity of lactic acid bacteria isolated from vacuum-packed meats. Journal of Applied Bacteriology 69, 302-310], (Weston et al., 1999) [Weston, R.J., Mitchell, K.R., Allen, K.L., 1999. Antibacterial phenolic components of New Zealand manuka honey. J. Food Chem. 64, 295-301] or disc diffusion (Taormina et al., 2001) [Taormina, Peter J., Niemira, Brendan A., Beuchat, Larry R., 2001. Inhibitory activity of honey against food home pathogens as influenced by the presence of hydrogen peroxide and level of antioxidant power. Int. J. Food Microbiol. 69, 217-225] assay using zones of inhibition as indicators of bacterial susceptibility. The development of a 24-h spectrophotometric assay employing 96-well microtiter plates, that is more sensitive and more amenable to statistical analysis than the assays currently employed, was undertaken. This simple and rapid assay permits extensive kinetic studies even in the presence of low honey concentrations, and is capable of detecting inhibitory levels below those recorded for well or disc diffusion assays. In this paper, we compare the assay to both well and disc diffusion assays. The results we obtained for the spectrophotometric method MIC values show that this method has greater sensitivity than the standard well and disc diffusion assays. In addition, inter- and intra-assay variance for this method was investigated, demonstrating the methods reproducibility and repeatability. (c) 2005 Elsevier B.V. All rights reserved.