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环境噬菌体对囊性纤维化患者铜绿假单胞菌菌株的体外和体内抗菌活性

In vitro and in vivo antibacterial activity of environmental bacteriophages against Pseudomonas aeruginosa strains from cystic fibrosis patients.

作者信息

Olszak Tomasz, Zarnowiec Paulina, Kaca Wieslaw, Danis-Wlodarczyk Katarzyna, Augustyniak Daria, Drevinek Pavel, de Soyza Anthony, McClean Siobhán, Drulis-Kawa Zuzanna

机构信息

Institute of Genetics and Microbiology, University of Wroclaw, Przybyszewskiego 63/77, 51-148, Wroclaw, Poland.

出版信息

Appl Microbiol Biotechnol. 2015 Jul;99(14):6021-33. doi: 10.1007/s00253-015-6492-6. Epub 2015 Mar 12.

Abstract

The goal of the study was to determine the relationship between in vitro/in vivo efficacy of environmental Pseudomonas phages and certain phenotypical properties of Pseudomonas aeruginosa (PA) strains. We studied the diversity between particular isolates and determined phage sensitivity in vitro and in vivo in the Galleria mellonella insect model. Twenty-eight lytic bacteriophages specific for PA were tested against 121 CF PA isolates including 29 mucoid PA strains. Most strains from cystic fibrosis (CF) patients were lysed by at least three phages (93.6 %), but completely insensitive strains were also present (6.4 %). Two phages PA5oct and KT28 exhibited high rates of lytic potency on 55-68 % of PA strains (72-86 % of mucoid isolates). We further explored phage activity against six PA strains (CF and non-CF) in vitro, comparing clonal differences in phage susceptibility with bacterial properties such as the ability to form biofilms, mucosity, twitching motility, and biochemical profiles. We observed the relationship between variation in phage susceptibility and Fourier transform infrared spectroscopy (FTIR) analysis in the spectra window of carbohydrates. The protective efficacy of two selected phages against PA PAO1 and 0038 infection was confirmed in vivo in G. mellonella larvae. Generally, the wax moth model results confirmed the data from in vitro assays, but in massive infection of CF isolates, the application of lytic phages probably led to the release of toxic compound causing an increase in larvae mortality. We assumed that apart of in vitro phage activity testing, a simple and convenient wax moth larvae model should be applied for the evaluation of in vivo effectiveness of particular phage preparations.

摘要

该研究的目的是确定环境假单胞菌噬菌体的体外/体内效力与铜绿假单胞菌(PA)菌株的某些表型特性之间的关系。我们研究了特定分离株之间的多样性,并在大蜡螟昆虫模型中测定了噬菌体的体外和体内敏感性。对28种特异性针对PA的裂解性噬菌体针对121株CF PA分离株进行了测试,其中包括29株黏液型PA菌株。大多数来自囊性纤维化(CF)患者的菌株被至少三种噬菌体裂解(93.6%),但也存在完全不敏感的菌株(6.4%)。两种噬菌体PA5oct和KT28对55 - 68%的PA菌株(72 - 86%的黏液型分离株)表现出高裂解效力。我们进一步在体外研究了针对六种PA菌株(CF和非CF)的噬菌体活性,比较了噬菌体敏感性的克隆差异与细菌特性,如形成生物膜的能力、黏液性、颤动运动性和生化谱。我们观察到在碳水化合物光谱窗口中噬菌体敏感性变化与傅里叶变换红外光谱(FTIR)分析之间的关系。在大蜡螟幼虫体内证实了两种选定噬菌体对PA PAO1和0038感染的保护效力。一般来说,蜡螟模型结果证实了体外试验的数据,但在CF分离株的大量感染中,裂解性噬菌体的应用可能导致有毒化合物的释放,从而导致幼虫死亡率增加。我们认为,除了体外噬菌体活性测试外,一个简单方便的蜡螟幼虫模型应用于评估特定噬菌体制剂的体内有效性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/894c/4480334/3c30726e6326/253_2015_6492_Fig1_HTML.jpg

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