Transmission Electronic Microscopy of Vibrio cholerae Biofilms on Chitin-Containing Substrates

S. Titova, I. Simonova, E. A. Men’shikova, V. S. Osadchaya
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Abstract

Introduction. The evolutionary association of Vibrio cholerae with chitin provided resistance to stress and protection from predators. The most important mechanism that provided V. cholerae with the effectiveness of association with chitin is biofilm formation. The ability to form a biofilm in V. cholerae depends on the presence of a factor, toxin-corrected adhesion pili (TCP), which are synthesized by the tcp A-F genes. One of the key methods for studying biofilms is microscopy. It allows one to visualize the structural elements and study various parameters of biofilms and the effects of various factors on them. Aim. To determine the epidemiological significance of the biofilm-forming ability of toxigenic strains by their morphological characteristics on chitin-containing substrates. Study of structural differences in biofilms of Vibrio cholerae tcpA+– and tcpA– strains on chitin-containing substrates. Results. It has been shown that Vibrio cholerae tcpA+– and tcpA– strains are able to form biofilms on the surface of chitin-containing substrates. The intensity of biofilm formation is more pronounced in tcpA+ strains, because V. cholerae ctxA+ tcpA+ cells in the biofilm are predominantly singly located and the surface of the chitinous exoskeleton with which they are in contact is intact, V. cholerae ctxA– tcpA– cells form chains in the biofilm, which indicates division processes, and scattered chitin of the endocuticle indicates activity of metabolic processes. Conclusion. The strains of V. cholerae used in the work, regardless of the presence or absence of the ctx and tcp genes, form bioplecs on a chitin substrate. The indicator of biofilm formation in terms of the thickness of the biofilm matrix is higher in V. cholerae ctxA+ tcpA+ , in terms of the degree of degradation of the chitin substrate it is higher in V. cholerae ctxA– tcpA– .
透射电子显微镜观察含几丁质基质上的霍乱弧菌生物膜
介绍。在进化过程中,霍乱弧菌与几丁质的结合提供了抗应激和抵御天敌的能力。使霍乱弧菌与几丁质有效结合的最重要机制是生物膜的形成。霍乱弧菌形成生物膜的能力取决于毒素校正粘附纤毛(TCP)因子的存在,TCP由tcp A-F基因合成。显微镜是研究生物膜的主要方法之一。通过显微镜可以观察生物膜的结构元素,研究生物膜的各种参数以及各种因素对生物膜的影响。目的通过致毒菌株在含几丁质基质上的形态特征,确定其生物膜形成能力的流行病学意义。研究霍乱弧菌 tcpA+- 和 tcpA- 菌株在含几丁质基质上形成生物膜的结构差异。结果。研究表明,霍乱弧菌 tcpA+- 和 tcpA- 菌株能在含几丁质的基质表面形成生物膜。tcpA+ 菌株形成生物膜的强度更明显,因为生物膜中的霍乱弧菌 ctxA+ tcpA+ 细胞主要是单个分布,与之接触的几丁质外骨骼表面完好无损;霍乱弧菌 ctxA- tcpA- 细胞在生物膜中形成链状,表明有分裂过程;内壳体的几丁质散落,表明有代谢过程。结论工作中使用的霍乱弧菌菌株,无论是否存在ctx和tcp基因,都能在几丁质基质上形成生物膜。从生物膜基质的厚度来看,霍乱弧菌 ctxA+ tcpA+ 的生物膜形成指标更高;从几丁质基质的降解程度来看,霍乱弧菌 ctxA- tcpA- 的生物膜形成指标更高。
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