丁香油对变形链球菌生物膜的影响

Study on the bacterial biofilm of Streptococcus pneumoniae by clove oil

目的 研究丁香油对变形链球菌细菌生物膜的影响.方法 通过气相色谱-质谱联用法确定丁香油的组成成分;制备变形链球菌悬液,进行杀菌实验,测定丁香油的最小抑菌浓度（minimum inhibitory concentration,MIC）及最小杀菌浓度（minimum bactericidal concentration,MBC）;利用激光共聚焦显微镜观察不同浓度的丁香油对变形链球菌生物膜形成的影响,并记录生物膜平均荧光强度.结果 通过气相色谱-质谱分析法确定8种丁香油成分,其中含量最高的为丁香酚,含量48.93%;其次为石竹烯、水杨酸甲酯,含量分别为20.78%、14.96%;丁香油对变形链球菌有抑菌作用,并随丁香油浓度增加而增强,当达到一定浓度时,可表现出杀菌作用,其MIC和MBC分别为0.3125%、2.5000%.丁香油对变形链球菌生物膜有抑制作用,随浓度增强抑制作用增强,当丁香油浓度2.5000%时,平均荧光强度为（0.082±0.007）,活菌比例明显降低,生物膜基本消失.结论 丁香油对变形链球菌及变形链球菌形成的生物膜有抑制及清除作用.

Objective To study the effect of clove oil on bacterial biofilm of Streptococcus pneumoniae. Methods The components of clove oil were determined by GC-MS mass spectrometry. The suspension of streptococcus mutans was prepared to carry out the germicidal test, and to determine the minimum inhibitory concentration （MIC） and the minimum bactericidal concentration （MBC） of clove oil. The effects of different concentrations of clove oil on the formation of biofilm were observed by laser confocal microscopy, and the average fluorescence intensity of live bacteria and dead bacteria in biofilm was recorded. Results Determination of clove oil 8 components were analyzed by GC-MS mass spectrometry method, most of the Eugenol content was 48.93%, followed by the Caryophyllene 20.78%, Methylis salicylas 14.96%. Through MIC, MBC and the experimental results showed that clove oil had antibacterial activity of Proteus, and with the increase of clove oil concentration, the inhibitory effect was better. When it reached a certain concentration, it can showed bactericidal effect that the MIC and MBC were 0.3125%, 0.6250%. The clove oil also had inhibitory effect on Streptococcus mutans biofilm with the concentration depedence. When the concentration of clove oil was 2.5000%, the average fluorescence intensity （live/dead bacteria） was 0.082 ± 0.007, the proportion of living bacteria decreased obviously, and biofilm disappeared. Conclusions The clove oil not only has inhibitory effect on Streptococcus mutants, but also can inhibit and clear the biofilm formation of Streptococcus.