Duarte Ana Catarina, Fernández Lucía, Jurado Andrea, Campelo Ana Belén, Shen Yang, Rodríguez Ana, García Pilar
Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Villaviciosa, Spain.
DairySafe Group, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain.
Front Microbiol. 2024 Aug 7;15:1438022. doi: 10.3389/fmicb.2024.1438022. eCollection 2024.
Bacteriophages have been shown to penetrate biofilms and replicate if they find suitable host cells. Therefore, these viruses appear to be a good option to tackle the biofilm problem and complement or even substitute more conventional antimicrobials. However, in order to successfully remove biofilms, in particular mature biofilms, phages may need to be administered along with other compounds. Phage-derived proteins, such as endolysins or depolymerases, offer a safer alternative to other compounds in the era of antibiotic resistance.
This study examined the interactions between phage with a polysaccharide depolymerase (Dpo7) from another phage () against biofilms formed by different strains, as determined by crystal violet staining, viable cell counts and microscopy analysis.
Our results demonstrated that there was synergy between the two antimicrobials, with a more significant decreased in biomass and viable cell number with the combination treatment compared to the phage and enzyme alone. This observation was confirmed by microscopy analysis, which also showed that polysaccharide depolymerase treatment reduced, but did not eliminate extracellular matrix polysaccharides. Activity assays on mutant strains did not identify teichoic acids or PNAG/PIA as the exclusive target of Dpo7, suggesting that may be both are degraded by this enzyme. Phage adsorption to cells was not significantly altered by incubation with Dpo7, indicating that the mechanism of the observed synergistic interaction is likely through loosening of the biofilm structure. This would allow easier access of the phage particles to their host cells and facilitate infection progression within the bacterial population.
噬菌体已被证明能够穿透生物膜,并且如果找到合适的宿主细胞就会进行复制。因此,这些病毒似乎是解决生物膜问题以及补充甚至替代更传统抗菌剂的一个很好的选择。然而,为了成功去除生物膜,特别是成熟的生物膜,噬菌体可能需要与其他化合物一起使用。在抗生素耐药性的时代,噬菌体衍生的蛋白质,如内溶素或解聚酶,提供了一种比其他化合物更安全的选择。
本研究通过结晶紫染色、活菌计数和显微镜分析,检测了一种噬菌体与另一种噬菌体()的多糖解聚酶(Dpo7)之间针对不同菌株形成的生物膜的相互作用。
我们的结果表明这两种抗菌剂之间存在协同作用,与单独使用噬菌体和酶相比,联合处理使生物量和活菌数量有更显著的减少。显微镜分析证实了这一观察结果,该分析还表明多糖解聚酶处理减少但并未消除细胞外基质多糖。对突变菌株的活性测定未确定磷壁酸或PNAG/PIA是Dpo7的唯一靶标,这表明两者可能都被该酶降解。与Dpo7孵育后,噬菌体对细胞的吸附没有显著改变,这表明观察到的协同相互作用机制可能是通过生物膜结构的松散。这将使噬菌体颗粒更容易接近其宿主细胞,并促进细菌群体内的感染进程。