Co-culture biofilm patterns among different Pseudomonas aeruginosa clones from cystic fibrosis patients

IF 4.9 Q1 MICROBIOLOGY
Irene Cadenas-Jiménez , Morten Levin Rybtke , Doaa Higazy , Sara Martí-Martí , Tim Tolker-Nielsen , Oana Ciofu , Niels Høiby
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引用次数: 0

Abstract

Background

Pseudomonas aeruginosa chronic lung infection is the leading cause of death in the cystic fibrosis (CF) population. The high genome versatility of this microorganism allows it to adapt to the hostile CF lung where the same clone can persist for decades. Paranasal sinuses serve as a reservoir for bacterial adaptation before lung infection. Our study investigates biofilm compatibility among identical and different P. aeruginosa genotypes from sinus and lungs of CF patients. Strains were further characterized by whole genome sequencing and motility assays were performed.

Methodology

Motility, gentamicin susceptibility and growth rates were assessed in four strains coming from three CF patients. The strains were subjected to whole genome sequencing with the Illumina MiSeq platform.
Conjugation assays using the mini Tn7 transposon were performed in order to tag bacteria with the fluorescent proteins YFP (yellow) and CFP (cyan). Biofilm experiments were carried out in a flow cell system and images were acquired using a confocal laser microscope (CLSM) on days 3 and 5. Four experiments were performed: Experiment 1 with two clonal isolates from sinus and lungs from patient P01 (CF430-142, CF430-11621); experiments 2 (CF430-11621 + 75885-B) and 3 (CF430-11621 + 80271-B) with two lung isolates belonging to two different clones from different patients (P02, P03) and experiment 4 with one lung strain (CF430-11621) and P. aeruginosa PAO1 reference strain.

Results

P. aeruginosa clonal isolates coming from paranasal sinuses and lungs from the same patient were able to form mixed biofilm. When different clones were employed no mixed biofilms were observed. Similar results were observed when combining the lung strain and the reference strain PAO1. Biofilms of both strains were observed in the flow-cell channels but no mixed biofilms of them were observed, with the exception of strain 75887-B which did not appear to form any biofilm when mixed with strain CF430-11621. All strains performed swarming while strains CF430-142 and 75887B lacked twitching motility. An aminoacidic change in SadB was observed in the strain 75887B.

Conclusion

Mixed biofilms were only observed when identical clones from the same patient were cultured together. Our experiments indicate that twitching motility does not significantly affect biofilm formation or architecture in our isolates.
囊性纤维化患者不同铜绿假单胞菌克隆共培养生物膜模式
背景:铜绿假单胞菌慢性肺部感染是囊性纤维化(CF)人群死亡的主要原因。这种微生物的高基因组多功能性使其能够适应敌对的CF肺,其中相同的克隆可以持续数十年。在肺部感染之前,鼻窦是细菌适应的储存库。本研究探讨了CF患者鼻窦和肺中相同和不同铜绿假单胞菌基因型的生物膜相容性。菌株进一步通过全基因组测序和运动性测定进行鉴定。方法对来自3例CF患者的4株菌株进行活力、庆大霉素敏感性和生长速率评估。利用Illumina MiSeq平台对菌株进行全基因组测序。使用mini Tn7转座子进行偶联实验,以便用荧光蛋白YFP(黄色)和CFP(青色)标记细菌。在流式细胞系统中进行生物膜实验,并在第3天和第5天使用共聚焦激光显微镜(CLSM)获取图像。实验1从患者P01 (CF430-142, CF430-11621)的鼻窦和肺中分离出2株克隆菌株;实验2 (CF430-11621 + 75885-B)和实验3 (CF430-11621 + 80271-B)分别分离了来自不同患者(P02、P03)的两个不同克隆的两株肺分离株,实验4分离了一株肺分离株(CF430-11621)和铜绿假单胞菌PAO1参考株。来自同一患者鼻窦和肺部的铜绿假单胞菌克隆分离株能够形成混合生物膜。使用不同的克隆时,未观察到混合的生物膜。将肺菌株与参考菌株PAO1联合使用时,观察到相似的结果。除菌株75887-B与菌株CF430-11621混合后未形成生物膜外,两种菌株在流细胞通道中均观察到生物膜,但未观察到混合生物膜。菌株CF430-142和75887B没有抽搐运动。在菌株75887B中观察到SadB的氨基酸变化。结论只有同一患者的相同克隆在一起培养时才会出现混合生物膜。我们的实验表明,抽搐运动不会显著影响我们分离物的生物膜形成或结构。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biofilm
Biofilm MICROBIOLOGY-
CiteScore
7.50
自引率
1.50%
发文量
30
审稿时长
57 days
期刊介绍:
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