超氧化物歧化酶在鸡白痢沙门氏菌耐受环丙沙星中的作用分析  

Effect of Superoxide Dismutase on Ciprofloxacin Tolerance of Salmonella Pullorum

作　　者：李艳莉 程伊洛 罗青平[2] 杨玉莹[1] 张腾飞[2] LI Yanli;CHENG Yiluo;LUO Qingping;YANG Yuying;ZHANG Tengfei(College of Animal Science,Yangtze University,Jingzhou,Hubei 434025;Key Laboratory of Animal Bacterial Disease Control Agent Development,Ministry of Agriculture and Rural Affairs,Hubei Key Laboratory of Pathogenic Microbiology of Livestock and Poultry,Institute of Animal Husbandry and Veterinary Medicine,Hubei Academy of Agricultural Sciences,Wuhan,Hubei 430064)

机构地区：[1]长江大学动物科学学院,湖北荆州434025 [2]湖北省农业科学院畜牧兽医研究所,农业农村部畜禽细菌病防治制剂创制重点实验室,畜禽病原微生物学湖北省重点实验室,湖北武汉430064

出　　处：《中国家禽》2024年第12期76-84,共9页China Poultry

基　　金：国家自然科学基金面上项目(32273014);湖北省自然科学基金创新群体项目(2021CFA019)。

摘　　要：为研究超氧化物歧化酶对鸡白痢沙门氏菌耐受环丙沙星的影响,试验利用λ-Red同源重组系统构建鸡白痢沙门氏菌C79-3的sod1、sod2、sod3和sod4的单基因缺失株和同为Cu/Zn家族的sod1与sod3、Fe/Mn家族的sod2与sod4的双基因缺失株及其互补株,比较分析亲本株、基因缺失株与互补株的生长曲线、生物被膜形成能力、过氧化氢和抗生素耐受能力的差异。结果显示:6个基因缺失株与亲本株的生长曲线无明显差别,但双基因缺失株Δsod1Δsod3和Δsod2Δsod4生物被膜形成能力显著下降(P<0.05),在环丙沙星处理后活性氧产生量显著高于亲本株(P<0.05);与亲本株、4个单基因缺失株和双基因缺失株Δsod1Δsod3相比,双基因缺失株Δsod2Δsod4在过氧化氢环境中存活率显著降低(P<0.01),且在环丙沙星处理后,其耐受能力大幅下降。以上结果表明以Fe和Mn作为辅因子的铁锰超氧化物歧化酶Sod2和Sod4通过协同作用,在鸡白痢沙门氏菌对环丙沙星的耐受中发挥关键作用,为进一步研究鸡白痢沙门氏菌耐药机制奠定理论基础。In order to understand the effect of superoxide dismutases on ciprofloxacin tolerance of Salmonella Pullorum,λ-Red homologous recombination system was used to construct the single gene deletion strains of sod1,sod2,sod3 and sod4,and the double gene deletion strains of sod1 and sod3 of Cu/Zn family and sod2 and sod4 of Fe/Mn family.Their complementary strains were also constructed in this study.Then,growth curve,biofilm formation ability,hydrogen peroxide and antibiotic tolerance were analyzed in this study.The results showed that there were no significant differences in growth curves among the 6 deletion strains with the parent strain,but the biofilm formation abilities ofΔsod1Δsod3 andΔsod2Δsod4 strains were significantly decreased(P<0.05),and reactive oxygen production of which by ciprofloxacin treatment was higher than that of the parent strain.Compared with the parent strain,the four single gene deletion strains andΔsod1Δsod3 double gene deletion strain,the survival rate ofΔsod2Δsod4 strain under hydrogen peroxide treatment was significantly reduced(P<0.05),and the ciprofloxacin tolerance ability of which was also significantly decreased(P<0.01).These results indicated that Fe-Mn superoxide dismutase Sod2 and Sod4 with Fe and Mn as cofactors played a key role in the resistance of Salmonella Pullorum to ciprofloxacin through synergistic interaction,which laid a theoretical foundation for further research on the mechanism of resistance of Salmonella Pullorum.

关 键 词：鸡白痢沙门氏菌 超氧化物歧化酶 过氧化氢耐受 环丙沙星耐受 

分 类 号：S855.1[农业科学—临床兽医学]