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目的:抗生素耐药性成为了全球性的健康问题。研究发现病原菌的多细胞行为在抗生素的耐药性中起着至关重要的作用(尤其是生物膜),因而通过抑制多细胞行为而控制耐药性成为当务之急。本文以奇异变形杆菌(Proteus mirabilis)为研究对象,考察它的发酵滤液对一种机会致病菌——铜绿假单胞菌(Pseudomonas aeruginosa)多细胞行为的作用,以期得到一株多细胞行为抑制菌:在不影响P.aeruginosa生长的前提下,抑制生物膜形成、EPS产生以及定向丛集运动,解除保护,减缓扩散,为降低P.aeruginosa耐药性,增强抗生素作用效果提供可能。方法:采用结晶紫生物膜测定法、蒽酮-硫酸法、平板检测法,探究P.mirabilis发酵滤液对P.aeruginosa生物膜、胞外多聚物、定向丛集运动和生长的影响。结果:P.mirabilis发酵滤液能显著抑制P.aeruginosa生物膜量,在体积百分比浓度为1%时,抑制率可达60.9%。该菌的发酵滤液还能阻碍P.aeruginosa的定向丛集运动,减弱它的吸附和扩散运动;同时,也减少了P.aeruginosa胞外多聚物的产量,在滤液体积百分比浓度为1%时,抑制率达到45.9%。更重要的是,固体平板实验证明该发酵滤液对P.aeruginosa的生长没有影响。结论:P.mirabilis在不影响病原菌生长的前提下,对病原菌的多细胞行为有一定的控制作用。其发酵滤液中存在着抑制微生物膜、定向丛集运动等的成分,在治疗细菌感染性疾病和降低抗生素耐药性方面有潜在应用价值。
Purpose: Antibiotic resistance has become a global health problem. The study found that multicellular behavior of pathogens plays a crucial role in antibiotic resistance (especially biofilms), and thus it is imperative to control drug resistance by inhibiting multicellular behavior. In this paper, Proteus mirabilis was studied to investigate the effect of its fermentation filtrate on the multicellular behavior of a opportunistic pathogen, Pseudomonas aeruginosa, in order to obtain a multicellular behavior inhibition Bacteria: inhibit the growth of P.aeruginosa, inhibit biofilm formation, EPS production and directional clustering movement, lifting the protection and slowing the spread, in order to reduce the resistance of P. aeruginosa and enhance the effect of antibiotics. Methods: The crystal violet biomembrane assay, anthrone-sulfuric acid assay and plate assay were used to investigate the effect of P.mirabilis ferment filtrate on P.aeruginosa biofilm, extracellular multimer, directional cluster movement and growth. Results: P.mirabilis ferment filtrate could significantly inhibit the amount of P.aeruginosa biofilm, and the inhibition rate could reach 60.9% when the volume percentage was 1%. The fermented filtrate of the bacterium could also hinder the directional clustering movement of P. aeruginosa and weaken the adsorption and diffusion of P.aeruginosa. At the same time, it also reduced the yield of P.aeruginosa extracellular multipolymer. When the filtrate concentration was 1% The inhibition rate reached 45.9%. More importantly, solid plate experiments demonstrated that the fermentation filtrate had no effect on the growth of P. aeruginosa. Conclusion: P.mirabilis can control the multicellular behavior of pathogenic bacteria under the premise of not affecting the growth of pathogenic bacteria. The fermentation filtrate contains components that inhibit the microbial membrane and directional cluster movement, which have potential application in the treatment of bacterial infectious diseases and the reduction of antibiotic resistance.