二氧化氯奶牛乳头消毒剂杀菌效果评价及杀菌机理研究

doi:10.16431/j.cnki.1671-7236.2021.07.040

摘要/Abstract

摘要： 试验旨在对本实验室研发的用于奶牛乳头消毒的二氧化氯消毒剂进行实验室杀菌效果评价，并对二氧化氯的杀菌机理进行初步研究。采用定量法杀菌试验对二氧化氯奶牛乳头消毒剂进行实验室杀菌效果评价；利用透射电子显微镜观察二氧化氯作用后金黄色葡萄球菌超微结构的改变；利用流式细胞仪测定二氧化氯作用后金黄色葡萄球菌细胞膜通透性的变化。定量法杀菌试验结果显示，当该消毒剂中二氧化氯的含量高于1.2 mg/L时，对无乳链球菌、铜绿假单胞菌的杀灭对数值高于3.00，即杀菌率高于99.9%；二氧化氯含量高于0.6 mg/L时，对金黄色葡萄球菌、大肠杆菌和白色念珠菌杀灭对数值高于3.00，即杀菌率高于99.9%；二氧化氯的含量为0.3 mg/L时，仍能有效杀灭表皮葡萄球菌、停乳链球菌；而二氧化氯含量高于28 mg/L，对于枯草芽孢杆菌的杀菌率才能达到100%。透射电子显微镜超微结构观察结果显示，100 mg/L二氧化氯作用于金黄色葡萄球菌5 min时，细胞膜出现了轻微的皱缩，细胞壁与胞膜之间出现了轻微的空隙，绝大多数细胞仍保持原有形态；作用15 min后，金黄色葡萄球菌形态发生了较明显的变化，细胞膜发生了明显的皱缩，细胞壁与细胞膜完全脱离，细胞质也出现了凝集。流式细胞仪检测金黄色葡萄球菌细胞膜通透性变化发现，二氧化氯处理后金黄色葡萄球菌细胞膜通透性发生了改变，其中含量为50 mg/L二氧化氯作用15 min后细胞膜通透性改变最明显。结合实验室杀菌效果评价结果发现，二氧化氯对细菌细胞膜通透性的改变仅仅是致死细菌的原因之一，二氧化氯对细菌的杀灭机理较复杂，仍需进一步的探索。

关键词: 二氧化氯; 乳头消毒剂; 杀菌效果评价; 杀菌机理

Abstract: The purpose of this experiment was to evaluate the germicidal efficacy of chlorine dioxide disinfectant developed by our laboratory for dairy teat disinfectant,and to preliminarily study the germicidal mechanism of chlorine dioxide.The germicidal efficacy of chlorine dioxide dairy teat disinfectant was evaluated by quantitative sterilization test.The ultrastructural changes of Staphylococcus aureus after chlorine dioxide treatment were observed by transmission electron microscope (TEM).The changes of cell membrane permeability of Staphylococcus aureus after chlorine dioxide treatment were determined by flow cytometry.The results of quantitative germicidal test showed that when the content of chlorine dioxide in the disinfectant was higher than 1.2 mg/L,the logarithm value of killing Streptococcus agalactiae and Pseudomonas aeruginosa was higher than 3.00,that was,the germicidal rate was higher than 99.9%.When the chlorine dioxide content was higher than 0.6 mg/L,the logarithm of killing Staphylococcus aureus,Escherichia coli and Candida albicans was higher than 3.00,that was,the killing rate was higher than 99.9%.When the content of chlorine dioxide was 0.3 mg/L,it could still effectively kill Staphylococcus epidermidis and Streptococcus dyslactiae.When the chlorine dioxide content was higher than 28 mg/L,the bactericidal rate of Bacillus subtilis could reach 100%.The results of TEM showed that the cell membrane of Staphylococcus aureus treated with 100 mg/L chlorine dioxide for 5 min showed slight shrinkage,and there was a slight gap between the cell wall and the cell membrane,and most of the cells still maintained their original morphology.After treated for 15 min,the morphology of Staphylococcus aureus changed obviously,the cell membrane collapsed obviously,the cell wall completely separated from the cell membrane,and the cytoplasm also agglutinated.Flow cytometry detection results showed that the membrane permeability of Staphylococcus aureus was changed after treated with chlorine dioxide,and the change of membrane permeability was the most obvious after treated with 50 mg/L chlorine dioxide for 15 min.Combined with the results of laboratory evaluation of germicidal efficacy,it was found that the change of cell membrane permeability was only of the causes of killing bacteria.The killing mechanism of chlorine dioxide on bacteria was more complex,and it still needed to be further explored.

Key words: chlorine dioxide; teat disinfectant; evaluation of germicidal efficacy; bactericidal mechanism

中图分类号:

S859.79⁺1

刘静, 徐飞, 张靖菊, 刘春双, 李秀波. 二氧化氯奶牛乳头消毒剂杀菌效果评价及杀菌机理研究[J]. 中国畜牧兽医, 2021, 48(7): 2635-2643.

LIU Jing, XU Fei, ZHANG Jingju, LIU Chunshuang, LI Xiubo. Evaluation of Germicidal Efficacy and Germicidal Mechanism of Chlorine Dioxide Teat Disinfectant[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2635-2643.

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