摘要
为了探究4-萜烯醇对沙门菌(Salmonella)的作用机制,首先通过刃天青指示剂测定4-萜烯醇对沙门菌的最小抑菌浓度(minimum inhibitory concentration,MIC),借助抑菌曲线评价其抗菌效果;其次通过扫描电子显微镜评价4-萜烯醇对沙门菌整体形态的影响;使用多功能酶标仪检测沙门菌胞内的ATP、碱性磷酸酶的泄露情况,探究4-萜烯醇对沙门菌细胞膜通透性的影响;最后通过核酸凝胶电泳分析4-萜烯醇对沙门菌DNA的作用。结果显示4-萜烯醇对沙门菌具有良好的体外抗菌效果,其MIC为0.016 mol/L;抑菌曲线证实1 MIC的4-萜烯醇至少在24 h内能够完全抑制沙门菌的生长;扫描电子显微镜观察到4-萜烯醇处理过的沙门菌表面变得粗糙、菌体皱缩、黏连;随着4-萜烯醇浓度的升高,沙门菌胞内ATP含量显著下降,碱性磷酸酶、核酸、蛋白质发生明显泄露,且呈现剂量依赖性;核酸凝胶结果显示4-萜烯醇对沙门菌的DNA没有结合及降解作用。本研究表明4-萜烯醇对沙门菌具有良好的体外抗菌效果,主要通过增加沙门菌细胞膜的通透性,使其内容物发生泄露、菌体皱缩变形等,从而抑制沙门菌。
To investigate the antibacterial mechanism of terpinen-4-ol against Salmonella,firstly,the minimum inhibitory concentration(MIC)of terpinen-4-ol against Salmonella was determined by resazurin indicator,and its antibacterial effect was evaluated by the antibacterial curve.Secondly,the effects of terpinene-4-ol on the overall morphology of Salmonella was observed by scanning electron microscope,and the leakage of ATP and alkaline phosphatase in Salmonella was detected to explore the effect of terpinen-4-ol on the membrane permeability of Salmonella using a multifunctional microplate reader.Finally,the effect of terpinen-4-ol on Salmonella DNA was analyzed by nucleic acid gel electrophoresis.The results showed that terpinen-4-ol had a good in vitro antibacterial effect on Salmonella,and its MIC was 0.016 mol/L.The antibacterial curve confirmed that 1 MIC terpinen-4-ol was able to completely inhibit the growth of Salmonella within at least 24h.It was observed by scanning electron microscope that the surface of Salmonella treated with terpinen-4-ol became rough,the cell shrunk and adhered.As terpinen-4-ol concentration increased,the intracellular ATP of Salmonella decreased significantly,and the alkaline phosphatase,nucleic acid,and protein were significantly leaked in a dose dependent manner.The result of nucleic acid gel showed that terpinen-4-ol did not bind and degrade Salmonella DNA.This study suggested that terpinen-4-ol had a good antibacterial effect on Salmonella in vitro,mainly by increasing the cell membrane permeability of Salmonella,causing its contents to leak,and shrinking and deforming the bacteria,thereby inhibiting Salmonella.
作者
连凯琪
孟蕊
王梦婷
张向丽
王国栋
张明亮
张坤朋
周玲玲
LIAN Kaiqi;MENG Rui;WANG Mengting;ZHANG Xiangli;WANG Guodong;ZHANG Mingliang;ZHANG Kunpeng;ZHOU Lingling(Henan Joint International Research Laboratory of Veterinary Biologics Researchand Application,Henan Engineering Technology Research Center of Medicinal Plant Comprehensive Utilization,Innovation and Practice Base for Postdoctors,Anyang Institute of Technology,Anyang,Henan 455000,China;College of Animal Science and Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第8期1684-1689,共6页
Chinese Journal of Veterinary Science
基金
国家自然科学基金青年资助项目(31802170)
河南省科技攻关计划资助项目(212102110090)
安阳工学院博士后启动基金资助项目(BHJ2020005)
安阳工学院教育教学改革研究与实践资助项目(AGJ2021077)。
关键词
4-萜烯醇
沙门菌
抑菌效果
细胞膜
泄露
terpinen-4-ol
Salmonella
antibacterial effect
cell membranes
leakage
