高铁酸钾对水体中黏菌素耐药基因mcr-1的作用  

Effect of Potassium Ferrate on Colistin Resistance Gene mcr-1 in Water

作　　者：王旺 王忠 吴俊杰 杨利利 邓芳 黎望 王立琦 WANG Wang;WANG Zhong;WU Junjie;YANG Lili;DENG Fang;LI Wang;WANG Liqi(Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau Mountainous Region,Ministry of Education,College of Animal Science,Guizhou University,Guiyang550025,China)

机构地区：[1]贵州大学/动物科学学院高原山地动物遗传育种与繁殖教育部重点实验室,贵州贵阳550025

出　　处：《畜牧与饲料科学》2024年第6期12-18,共7页Animal Husbandry and Feed Science

基　　金：国家自然科学基金项目(32160853,32260829);贵州省自然科学基金项目(黔科合基础-ZK[2024]一般040,黔科合基础[2022]一般113);贵州大学自然科学专项(特岗)科研基金项目([2022]21号)。

摘　　要：[目的]探讨高铁酸钾对水体中质粒介导的黏菌素耐药基因mcr-1的作用。[方法]采用4因素3水平正交试验法,通过实时荧光定量PCR测定水体中mcr-1基因的绝对丰度,以mcr-1基因消减率为指标,考察不同因素及水平[高铁酸钾(A):7.09、14.17、21.26 mg/L;反应温度(B):40、50、60℃;反应时间(C):5、15、30 min;反应pH值(D):5、7、9)]对mcr-1基因拷贝数的影响。[结果]4个因素中,对mcr-1基因绝对丰度影响最大的是高铁酸钾用量,其次是反应pH值和反应时间,影响最小的是反应温度;对mcr-1基因消减效果最佳的水平组合为A_(3)B_(3)C_(1)D_(3),即采用21.26 mg/L高铁酸钾(以Fe计为6 mg/L),在反应pH值为5、反应温度为60℃的条件下处理15 min,该条件下mcr-1基因的消减率为55%。但在某些因素水平组合下,mcr-1基因会出现拷贝数增加的现象。[结论]高铁酸钾可通过改变DNA浓度来影响mcr-1基因含量。在适当条件下,高铁酸钾可消减水体中的mcr-1基因;但在某些因素水平组合下,mcr-1基因复制增加。[Objective]The aim of this study was to investigate the effect of potassium ferrate on plasmid-mediated colistin resistance gene mcr-1.[Method]The absolute abundance of mcr-1 gene in water was determined by real-time fluorescence quantitative PCR using four-factor and three-level orthogonal method.The elimination rate of mcr-1 gene was used as an index to investigate different factors and levels(potassium ferrate:7.09 mg/L,14.17 mg/L,21.26 mg/L;temperature:40℃,50℃,60℃;reaction time:5 min,15 min,30 min;pH:5,7,9)on mcr-1 gene content.[Result]Among the four factors,the greatest influence on the absolute abundance of mcr-1 gene was the dosage of potassium ferrate,followed by the reaction pH and reaction time,and the least influence was the reaction temperature.The level combination with the best effect on mcr-1 gene ablation was A_(3)B_(3)C_(1)D_(3),which was treated with 21.26 mg/L potassium ferrate(6 mg/L in Fe)for 15 min at a reaction pH of 5 and a reaction temperature of 60℃,and the ablation rate of the mcr-1 gene under this condition was 55%.However,the copy number of mcr-1 gene increased under certain combinations of factors.[Conclusion]Potassium ferrate could affect mcr-1 gene levels by altering DNA concentrations.Under appropriate conditions,potassium ferrate abrogated the mcr-1 gene in the water.However,under certain combinations of factor levels,mcr-1 gene replication will increase.

关 键 词：mcr-1 高铁酸钾 耐药基因 正交试验 环境 

分 类 号：X703[环境科学与工程—环境工程] X172[农业科学—临床兽医学] S859.796[农业科学—兽医学]