Adhesiveness of the specific phages to yersiniae cells assessed by optical trapping

I. Konyshev, L. Dudina, N. A. Morozova, A. Byvalov
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Abstract

In recent years, there has been increasing interest in using bacteriophages in the complex therapy of some infectious diseases. Objectively, it is related to the spread of antibiotic resistance and a small number of side effects triggered by phages applied locally and parenterally. Yersiniae are no exception to this rule, but their interaction with virions has not been studied thoroughly. In three types of lytic Yersiniae bacteriophages are used for diagnostic purposes, which differ in specificity, virulence, and morphological features. We evaluated the interaction force between phages and bacteriae by optical trapping. Materials and methods. Live Yersinia pseudotuberculosis and Y. pestis cells with different LPS structure and preparations of three bacteriophages Pokrovskayas phage, dHerelles phage (also named as the pseudotuberculosis phage) and the phage L-413C were used. The Pokrovskayas phage is well studied and widely used in diagnostics, but the mechanism of its adsorption on Y. pestis and Y. pseudotuberculosis cells is not well-understood. The phage L-413C lyses plague bacteria, while dHerrels phage can lyse both pathogens. Virions were linked to the surface of aminated glass. Optically trapped microbial cells were approached to the glass surface and then retracted in the opposite direction. The amplitude of the leap was recalculated into force through calibration coefficients. The primary data were processed in the program R; statistical analysis was performed using Matlab 7.0 and Statistica 12 Results and discussion. The analysis of histograms revealed the presence of a specific component in the Y. pseudotuberculosisdHerrells phage (Fmean = 7.463.52 pN) system, as well as in all pairs involving plague microbe. In the latter case, the interaction forces were comparable: 8.643.83 pN for dHerelles phage,11.034.22 pN for Pokrovskayas phage, and 10.424.79 pN for phage L-413C. The average detachment force from the substrate treated with bovine serum albumin (BSA) was comparable for both cell types. Our results convincingly prove an opportunity for using the optical trapping to estimate interaction force in other bacteriumphage systems.
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用光学诱捕法评估特定噬菌体对耶尔森氏菌细胞的粘附性
近年来,利用噬菌体进行感染性疾病的综合治疗已引起人们越来越多的兴趣。客观上,它与抗生素耐药性的传播以及局部和非肠道应用噬菌体引发的少量副作用有关。耶尔森尼菌也不例外,但它们与病毒粒子的相互作用还没有得到彻底的研究。在三种类型的裂解耶尔森菌噬菌体用于诊断目的,其特异性,毒力和形态特征不同。我们用光学捕获法评估了噬菌体与细菌之间的相互作用力。材料和方法。采用不同LPS结构的假结核耶尔森氏菌和鼠疫耶尔森氏菌活细胞,制备三种噬菌体Pokrovskayas噬菌体、dHerelles噬菌体(又称假结核噬菌体)和噬菌体L-413C。Pokrovskayas噬菌体在诊断中得到了广泛的研究和应用,但其吸附鼠疫菌和假结核杆菌细胞的机制尚不清楚。噬菌体L-413C可以裂解鼠疫菌,而大肠杆菌噬菌体可以裂解这两种病原体。病毒粒子被连接到胺化玻璃的表面。光学捕获的微生物细胞靠近玻璃表面,然后向相反方向缩回。通过校正系数将跳跃幅度重新计算为力。原始数据在R程序中处理;使用Matlab 7.0和Statistica 12进行统计分析。直方图分析显示,假结核杆菌-赫氏噬菌体(Fmean = 7.463.52 pN)系统以及所有涉及鼠疫微生物的噬菌体对中存在一种特定成分。后一种情况下,相互作用力具有可比性:dHerelles噬菌体为8.643.83 pN, Pokrovskayas噬菌体为11.034.22 pN, L-413C噬菌体为10.424.79 pN。用牛血清白蛋白(BSA)处理的底物的平均脱离力对两种细胞类型是相当的。我们的结果令人信服地证明了利用光学捕获来估计其他噬菌体系统中的相互作用力的机会。
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