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铜绿假单胞菌小菌落变异体在抑制烟曲霉生物膜方面表现出异质性。

Small Colony Variants of Pseudomonas aeruginosa Display Heterogeneity in Inhibiting Aspergillus fumigatus Biofilm.

机构信息

California Institute for Medical Research, 2260 Clove Dr., San Jose, CA, 95128, USA.

Division of Infectious Disease and Geographic Medicine, Department of Medicine, Stanford School of Medicine, Stanford University, Stanford, CA, 94305, USA.

出版信息

Mycopathologia. 2018 Feb;183(1):263-272. doi: 10.1007/s11046-017-0186-9. Epub 2017 Aug 7.

Abstract

Pseudomonas aeruginosa and Aspergillus fumigatus are major microbes in cystic fibrosis (CF). We reported non-mucoid P. aeruginosa isolates more inhibitory to A. fumigatus than mucoid ones. Another CF P. aeruginosa phenotype, small colony variants (SCVs), is an unknown factor in intermicrobial competition with A. fumigatus. Clinical SCV isolates and reference CF non-mucoid isolate (Pa10, producing normal-sized colonies) were compared. Live cells of P. aeruginosa or filtrates from P. aeruginosa planktonic or biofilm cultures were co-incubated with A. fumigatus growing under conditions allowing biofilm formation or with preformed biofilm. Metabolic activity of A. fumigatus biofilm was then measured. When necessary, assays were done after adjustment for growth differences by adding fresh medium to the planktonic culture filtrate. Pyoverdine determinations were performed spectrophotometrically on the planktonic culture filtrates. In all experimental conditions (live cells and planktonic or biofilm culture filtrates of P. aeruginosa versus A. fumigatus biofilm formation or preformed biofilm), three SCV isolates were less inhibitory than Pa10, two equal or more inhibitory. Adjusting planktonic culture filtrates for growth differences showed SCV inhibition differences variably related to growth or deficient inhibitor production. Studies suggested the principal P. aeruginosa inhibitor to be pyoverdine. SCV isolates appear heterogeneous in their capacity to inhibit A. fumigatus biofilm. SCV isolates can be important in the CF microbiome, because they are capable of intermicrobial inhibition.

摘要

铜绿假单胞菌和烟曲霉是囊性纤维化(CF)的主要微生物。我们报道了非粘液型铜绿假单胞菌分离株比粘液型铜绿假单胞菌对烟曲霉的抑制作用更强。CF 铜绿假单胞菌的另一种表型,小菌落变异体(SCV),是与烟曲霉进行微生物间竞争的未知因素。比较了临床 SCV 分离株和参考 CF 非粘液型分离株(Pa10,产生正常大小的菌落)。将铜绿假单胞菌的活细胞或来自铜绿假单胞菌浮游或生物膜培养物的滤液与在允许生物膜形成的条件下生长的烟曲霉或预先形成的生物膜一起孵育。然后测量烟曲霉生物膜的代谢活性。必要时,通过向浮游培养物滤液中添加新鲜培养基来调整生长差异,对测定进行调整。在所有实验条件下(铜绿假单胞菌的活细胞和浮游或生物膜培养物滤液与烟曲霉生物膜形成或预先形成的生物膜相比),三个 SCV 分离株的抑制作用小于 Pa10,两个分离株的抑制作用相等或更强。调整浮游培养物滤液以适应生长差异表明,SCV 抑制差异与生长或缺乏抑制剂产生有关。研究表明,铜绿假单胞菌的主要抑制剂是绿脓菌素。SCV 分离株在抑制烟曲霉生物膜的能力上表现出异质性。SCV 分离株在 CF 微生物组中可能很重要,因为它们能够进行微生物间抑制。

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