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论文摘要

林麝化脓病病原菌化脓隐秘杆菌与铜绿假单胞菌互作机制研究

Interspecific Interaction Between <i>Pseudomonas aeruginosa</i> and <i>Trueperella pyogenes</i> From the Abscesses Disease of <i>Moschus berezovskii</i>

作者:袁阳, 李静, 张爱雪, 林家富, 褚以文, 王欣荣*, 赵克雷*

Author:抗生素研究与再评价四川省重点实验室, 四川抗菌素工业研究所, 成都大学, 成都 610052

收稿日期:2020-01-08          年卷(期)页码:2020,39(3): 241-248,():-

期刊名称:四川动物

Journal Name:Sichuan Journal of Zoology

关键字:林麝;化脓病;化脓隐秘杆菌;铜绿假单胞菌;基因敲除;种间相互作用

Key words:Moschus berezovskii; abscess disease; Trueperella pyogenes; Pseudomonas aeruginosa; gene knock-out; interspecific interaction

基金项目:四川省科学技术协会青年人才托举工程项目(2018RCTJ);重大新药创制项目(2018ZX09711-001)

中文摘要

化脓隐秘杆菌Trueperella pyogenes是我国Ⅰ级重点保护野生动物林麝Moschus berezovskii的化脓病原发病原菌,但在化脓病后期的病灶中常能检测到大量铜绿假单胞菌Pseudomonas aeruginosa。虽然在病灶中能同时分离到这2种病原菌,但是它们的种间关系以及优势菌转换机制很大程度上未知。本研究通过构建多种铜绿假单胞菌群体感应基因敲除菌株并结合平板距离培养实验,探讨了化脓隐秘杆菌与铜绿假单胞菌的种间互作关系。结果发现,野生型铜绿假单胞菌的胞外产物可以显著抑制化脓隐秘杆菌的生长;不同铜绿假单胞菌群体感应关键基因缺失菌株均表现出对化脓隐秘杆菌不同程度的抑制作用,但与单突变菌株相比,lasR和rhlR双突变菌株对化脓隐秘杆菌的抑制作用明显减弱。这些发现表明,铜绿假单胞菌可以通过群体感应系统介导的胞外产物来提高对化脓隐秘杆菌的竞争优势。因此,本研究为林麝化脓病过程中的优势病原菌的替换和病情恶化甚至死亡提供了合理的解释,也有助于进一步提高对林麝化脓病病理的认识、治疗方案的改进和新型抗感染药物的研发。

英文摘要

Trueperella pyogenesis the primary pathogen of the abscess disease of forest musk deer (Moschus berezovskii), but a large amount ofPseudomonas aeruginosacan usually be detected in the purulent lesions of dying or dead deer. Although these 2 pathogenic bacteria can be co-isolated, their interaction relationship and the transition mechanism of predominant bacteria in abscess lesions are largely unknown. In this study, the interspecific interaction ofP. aeruginosawithT. pyogeneswas explored by constructing different mutant strains inP. aeruginosaquorum-sensing (QS) system and performing a series of detailedin vitroproximity assays. The results showed that the extracellular product ofP. aeruginosacould significantly inhibit the growth of neighboringT. pyogenes. In comparison to the wild-typeP. aeruginosa, knock-out of any core regulatory genes ofP. aeruginosaQS system would impair the inhibition ofP. aeruginosaonT. pyogenes, albeit the growth ofT. pyogenescould still be blocked by the presence of the extracellular products ofP. aeruginosaQS mutants. Notably, the double mutant strain PAO1-ΔlasRrhlRhad the slightest inhibitory effect onT. pyogenes. These findings suggested thatP. aeruginosahad significant competitive advantage overT. pyogenesby producing various QS-controlled extracellular products. Therefore, the current study provides a reasonable explanation for the transition of dominant bacterial species during the development of forest musk deer abscess diseases, contributes to further understanding the pathology of forest musk deer abscess diseases, the improvement of treatment options and the development of novel anti-infective drugs.

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