芽胞抗力快速弱化技术研究

Rapid Weakening Techniques of Germ Resistance

以L-丙氨酸缓冲液为发芽剂,结合芬顿反应原理,观察发芽-氧化损伤效应对芽胞的杀灭效果,以期为新型炭疽疫源地净化方法的深入研究奠定基础。以腊样芽胞为试验菌,采用透射电镜、激光扫描共聚焦显微镜、活菌计数等方法观察芽胞发芽过程的超微结构、核酸含量变化,以及在芬顿反应的联合作用下发芽体的活性变化。在20~30 min的发芽过程中,芽胞核心密度降低,核心与皮质、皮质与外壁之间界限模糊,芽胞外壁和芽胞衣有破裂,通透性增加,进一步有皮质消失、细胞核与细胞质融合、细胞膜基本形成的现象;发芽体荧光强度不断增加,显示菌体中核酸的活性和含量不断增加;发芽体对化学因子的抗力明显下降,H2O2浓度为0.20 mol/L的Fenton反应系统作用60 min时,发芽体灭活可达到3.016个对数级。诱导发芽和反应的联合处理程序可显著提高芽胞的灭活水平。

Based on the germination agent of L-alanine buffer combined with the principle of Fendun reaction,the killing result of germination-oxidation traumatic effects against germ were observed so as to lay down foundation of studying deeply on cleaning methods of the epidemic source new type of anthrax.The germ of Bacillus cereus was used as a test bacteria and observed its super microstructure of its germination process,content changes of nucleic acid through transmission electron microscope,laser scanning confocal microscope,living cell counting etc.,as well as the activity changes of germinant body under the united effect of Fenton method.The results showed that during the germinant process in 20~30 min.the density of core were decreased and the margins between core and cortex,the bounds cortex and exospore were blurred;and there were some break at the exospore and coat,the permeability increased,some cortex vanished,nuclei and protoplasm fused,and cell membrane basically formed,and fluorescence intensity germination body continuously increased,indicating the activity and content of nucleic acid of thallus continuously increased;the resistance of germination body against chemical factor obviously decreased.After treated by Fenton reaction（0.2 mol H2O2） for 60 min,the inactivation of germination body was as high as 3.016 logarithmic level.Therefore,joint treatment procedure of germination with reaction could obviously increase the inactivation level of germs.