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利用新分离的噬菌体控制铜绿假单胞菌 PAO1 和 ATCC 10145 生物膜。

Use of newly isolated phages for control of Pseudomonas aeruginosa PAO1 and ATCC 10145 biofilms.

机构信息

Institute of Biotechnology and Bioengineering, Center of Biological Engineering, University of Minho, Campus de Gualtar 4710-057, Braga, Portugal.

出版信息

Res Microbiol. 2011 Oct;162(8):798-806. doi: 10.1016/j.resmic.2011.06.010. Epub 2011 Jul 5.

Abstract

Pseudomonas aeruginosa is a relevant opportunistic pathogen involved in nosocomial infections that frequently shows low antibiotic susceptibility. One of its virulence factors is associated with the ability to adhere to surfaces and form virulent biofilms. This work describes the isolation and characterization of lytic phages capable of infecting antibiotic-resistant P. aeruginosa strains. In addition, characterization of P. aeruginosa biofilms and the potential of newly isolated phages for planktonic and biofilm control was accessed. According to the results, the isolated phages showed different spectra of activity and efficiency of lysis. Four broad lytic phages were selected for infection of planktonic cells; however, despite their broad range of activity, two of the selected phages failed to efficiently control planktonic cultures. Therefore, only two phages (phiIBB-PAA2 and phiIBB-PAP21), highly capable of causing strong biomass reduction of planktonic cells, were tested against 24 h biofilms using a m.o.i. of 1. Both phages reduced approximately 1-2 log the biofilm population after 2 h of infection and reduction was further enhanced after 6 h of biofilm infection. However, biofilm cells of P. aeruginosa PAO1 acquired resistance to phiIBB-PAP21; consequently, an increase in the number of cells after 24 h of treatment was observed. Conversely, phage phiIB-PAA2 for P. aeruginosa ATCC10145 continued to destroy biofilm cells, even after 24 h of infection. In these biofilms, phages caused a 3 log reduction in the number of viable counts of biofilm cells.

摘要

铜绿假单胞菌是一种与医院感染相关的机会性病原体,其对抗生素的敏感性通常较低。其毒力因子之一与黏附表面和形成毒性生物膜的能力有关。本工作描述了能够感染抗生素耐药铜绿假单胞菌菌株的裂解噬菌体的分离和鉴定。此外,还对铜绿假单胞菌生物膜的特性和新分离噬菌体对浮游生物和生物膜控制的潜力进行了评估。结果表明,分离的噬菌体显示出不同的活性谱和裂解效率。选择了 4 种广谱裂解噬菌体感染浮游细胞;然而,尽管它们的活性范围很广,但所选的 2 种噬菌体未能有效地控制浮游培养物。因此,仅选择了 2 种噬菌体能有效地引起浮游细胞强烈的生物量减少,并用 m.o.i.为 1 测试它们对 24 小时生物膜的影响。在感染后 2 小时内,两种噬菌体均可使生物膜的群体减少约 1-2 个对数级,在感染生物膜 6 小时后,减少效果进一步增强。然而,铜绿假单胞菌 PAO1 的生物膜细胞对 phiIBB-PAP21 产生了抗性;因此,在处理 24 小时后观察到细胞数量增加。相反,phiIB-PAA2 噬菌体对铜绿假单胞菌 ATCC10145 的生物膜细胞继续造成破坏,即使在感染 24 小时后也是如此。在这些生物膜中,噬菌体使生物膜细胞的活菌数减少了 3 个对数级。

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