Possible mechanisms of the destruction of spores of the vaccine strain Bacillus anthracis STI-1 under the influence of alcohol sols of metal nanoparticles

G. Frolov, N. V. Zavyalova, I. Lundovskikh, M. R. Shabalina, I. P. Pogorelskiy, K. I. Gurin, A. V. Mironin
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Abstract

The results are presented of a comparative study of the sporicidal activity of alcohol sols of nanoparticles of titanium, copper, zinc and tantalum metals in the course of their interaction with a rehydrated lyophilized culture of spores of the vaccine strain Bacillus anthracis STI-1 with the loss of the ability of spores to germinate in nutrient media under optimal conditions using bacteriology and electronic microscopy. The most pronounced sporicidal effect is observed when titanium alcohol sol is exposed to spores. At the same time, spores of B. anthracis STI-1 in titanium alcohol sol are affected by ethyl alcohol, which causes dehydration and denaturation of the protein, the antiseptic cetylpyridinium chloride as a surfactant, which is also a strong electrophilic agent that destroys the exosporium and spore shells, and titanium nanoparticles, which, due to the high surface energy when exposed to spores, cause their pronounced massive adhesion. In addition, titanium nanoparticles, the zeta (? -) value of the potential of which is -44.5 mV, cause a stable energy state of the dispersed system of alcohol sol, which acts as a biocatalyst for intracellular enzymes, causing the hydrolysis of polymer structures of the spore shells and cortex, as well as destroyed nucleoid – an area of spores containing structured genetic material. Spores that do not have a nucleoid in electron microscopy acquire the form of "shadows". As a result of the action of titanium alcohol sol, the activation of germination and the formation of the so-called "metabolic" spores, from which the vegetative cells of B. anthracis STI-1 should have been formed, are prevented. The reason for the high sporicidal efficiency of the alcohol sol of titanium nanoparticles is associated both with the structure and properties and with the technology of synthesis of titanium nanoparticles in the low-temperature plasma zone with the formation, due to the excess of the surfactant cetylpyridinium chloride, of a two-layer shell around the metal nanoparticles, which leads to their "conservation" until the moment of contact with spores and a sharp slowdown in their oxidation in the aquatic environment. The destruction of the formed two-layer shell of metal nanoparticles begins with contact with spores with the simultaneous onset of the formation of titanium ions from the phase of nanoparticles, which actively destroy chemically the shells of the spores. Keywords: microorganisms, Bacillus anthracis STI-1, spores, metal nanoparticles, zeta potential, disinfectant compositions, sporicidal activity.
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金属纳米颗粒酒精溶胶对炭疽芽孢杆菌STI-1疫苗株孢子破坏的可能机制
采用细菌学和电子显微镜对钛、铜、锌和钽金属纳米颗粒的酒精溶胶与炭疽芽孢杆菌STI-1疫苗菌株的再水合冻干培养物相互作用过程中的杀孢活性进行了比较研究,结果表明孢子在最佳条件下在营养培养基中发芽的能力丧失。当钛醇溶胶暴露于孢子时,观察到最明显的杀孢效果。同时,钛醇溶胶中的炭疽芽孢杆菌STI-1孢子受到乙醇的影响,乙醇会导致蛋白质脱水变性,防腐剂氯化十六烷基吡啶作为表面活性剂,也是一种强亲电剂,可以破坏外孢子和孢子壳,钛纳米颗粒由于暴露于孢子时具有高表面能,导致其明显的大量粘附。此外,钛纳米粒子、zeta (?-)值,其电位为-44.5 mV,导致醇溶胶分散体系处于稳定的能量状态,醇溶胶作为细胞内酶的生物催化剂,导致孢子壳和皮层的聚合物结构水解,以及破坏类核——孢子中含有结构遗传物质的区域。在电子显微镜下,没有类核的孢子获得“阴影”的形式。由于钛醇溶胶的作用,激活萌发和形成所谓的“代谢”孢子(炭疽芽孢杆菌STI-1的营养细胞本应由这些孢子形成)被阻止了。钛纳米颗粒醇溶胶的高杀孢效率的原因与钛纳米颗粒的结构和性质以及在低温等离子体区合成钛纳米颗粒的技术有关,由于表面活性剂十六烷基吡啶的过量,在金属纳米颗粒周围形成了两层外壳。这导致了它们的“保存”,直到与孢子接触的那一刻,以及它们在水生环境中氧化的急剧减缓。形成的两层金属纳米颗粒外壳的破坏开始于与孢子接触,同时开始从纳米颗粒相形成钛离子,它们积极地化学破坏孢子的外壳。关键词:微生物,炭疽芽孢杆菌STI-1,孢子,金属纳米颗粒,zeta电位,消毒剂成分,杀孢活性
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