Molecular genetic basis of biofilm formation as a component of Vibrio Cholerae persistence in water reservoirs of Russian Federation

Aim. In this study, we set out to identify the homologues of genes from the msh-cluster in the genomes of non-toxigenic V. cholerae, to perform the bioinformatics analysis of their products, as well as to study the adhesive properties of strains containing altered genes. Materials and methods. We analysed 17 clinical strains of non-O1/non-O139 V. cholerae and 2 strains of the O1 serogroup isolated from water bodies. Genes belonging to the msh-cluster were identified in the whole genomes using the BLASTN 2.2.29 and BioEdit 7.2.5 programs. Gene translation, comparative analysis of their nucleotide sequences and the amino acid sequences of deduced products were performed using the Vector NTI Advance 11 (Invitrogen). Results and discusssion. In 18 out of the 19 studied genomes we identified gene clusters responsible for production of adhesion pili (mshH-Q) represented by diverse alleles, the majority of which differed from the prototype genes of the msh-cluster in nucleotide composition but had the same localization and arrangement. Only one strain had a cluster that was close to that of the prototype. A bioinformatics analysis of their deduced products indicated that the amino acid sequence of the major MshA pilus subunit is homologous to the prototype only in a short N-terminal region (1-41) while sharing no similarities with the rest of the sequence. Nevertheless, this protein, similar to VcfA described by Kuroki H. et al. (2001) and designated by us as MshA-like, retained a putative pilus domain. A similar pattern was observed in the minor subunits designated as MshC-like. Other minor subunits also retained their characteristic domains. All of the strains agglutinated human erythrocytes (group O) and chicken erythrocytes, and in isolates harboring modified mshA-like and mshC-like genes the reaction was not inhibited by mannose. Since most of the studied strains were isolated from hospitalized patients, it is possible that in non-toxigenic V. cholerae lacking the pathogenicity island VPI, MSHA-like pili may serve as a colonization factor of the human intestine, in contrast to VPI-positive strains. The obtained information provides a basis for experimental verification of this assumption. This article was translated by Natalia Popova Laboratory of Scientific Translation. е.В. монахова, и.В. архангельская, с.В. Титова, р.В. Писанов MSHA-Подобные Пили нетокСигенных штаммоВ холерных ВибрионоВФКУЗ «Ростовский-на-Дону научно-исследовательский противочумный институт», Ростов-на-Дону, Российская Федерация цель исследования состояла в идентификации гомологов генов msh-кластера в геномах нетоксигенных холерных вибрионов, биоинформационном анализе их продуктов и изучении адгезивных свойств штаммов, содержащих измененные гены. материалы и методы. в работе использовано 17 клинических штаммов холерных вибрионов не о1/не о139 и 2 штамма о1 серогруппы, выделенные из водоемов. гены msh-кластера идентифицировали в полных геномах с помощью программ BLASTN 2.2.29 и BioEdit 7.2.5; трансляцию генов, сравнительный анализ их нуклеотидных...

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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MSHA-like pili of non-toxigenic Vibrio cholerae strains

Монахова

Архангельская

Титова

et al. 2019

“…полногеномное секвенирование на платформе MiSeq (Illumina) и сборку геномов выполняли, как описано ранее [3], кластерный анализ полученных сиквенсов (WGSs, whole genome sequences) проводили согласно предложенной ранее схеме [6 N o t e s : n/N -number of sequenced strains / total number of isolates; pt -prototype genes. Number of SNPs is determined against V. chole rae N16961 prototype genes.…”

Section: материалы и методыunclassified

“…иногда регистрируются случаи выделения практически идентичных штаммов из одних и тех же источников в диапазоне от трех недель до двух месяцев, и за это время может произойти заражение людей при условии наличия у вибрионов патогенетического потенциала. ретроспективный анализ генотипов показал, что чаще всего клональные комплексы образуют штаммы, содержащие остров патогенности VPI с генами токсин-корегулируемых пилей TCP, тогда как CTX -VPIотличаются более выраженной гетерогенностью, хотя некоторые тоже обладают клональностью [2][3][4][5]. так, в 2021 г. в ростове-на-дону был выявлен клональный комплекс, включающий 10 CTX -VPI + изолятов, которые обнаруживались в воде р. дон в трех разных точках отбора в течение трех недель.…”

unclassified

Genotypic Characteristics of CTX–VPI<sup>+</sup> Clonal Complexes of <i>Vibrio cholerae</i> O1 Found in Water Bodies of the Rostov Region

Monakhova,

Noskov,

Kruglikov

et al. 2023

The aim of the work was to identify genotypic features of clonal complexes formed by CTX–VPI+ strains of Vibrio cholerae isolated from water bodies of the Rostov Region in different years, based on bioinformatics analysis of their whole genome sequences (WGSs).Materials and methods. Whole genome sequencing was performed on the MiSeq (Illumina) platform; identification of genetic determinants in WGSs and bioinformatics analysis – by means of BioEdit, BLASTN, BLASTP, CARD, Vector NTI software packages.Results and discussion. The strains were almost identical in each group of 2002-, 2005-, 2007-, 2020-, 2021-, and 2022-isolates and were found at different points of water sampling over a longer or shorter periods of time (from 3 weeks to 2 months). Bioinformatics analysis of WGSs of representative strains divided them into 4 clusters, which can be considered as separate clonal complexes sharing a number of characteristic genetic markers: 1) 2002-isolates containing one SNP in regulatory rpoS gene; 2) 2007-strains carrying tandemly duplicated preCTX prophage; 3) strains isolated in 2020 in Rostov-on-Don and in 2005, during epidemiological complications in the Kamensk district of the Rostov Region, which had a “hybrid” pathogenicity island including the cluster of type 3 secretion system and the nan-nag region of the VPI-2 island, and 9 SNPs in rpoS gene; 4) 2021- and 2022-strains and a single 2018-isolate containing a 9 bp deletion in vchC collagenase gene. Despite the revealed differences, representatives of all clonal complexes possessed sufficient sets of intact determinants of pathogenicity/persistence factors to realize virulent properties and ability to survive in water bodies. Detection of clonal complexes of potentially pathogenic strains in surface water bodies indicates occasional emergence of favorable conditions for their long-term circulation, which emphasizes the importance of constant monitoring of V. cholerae in the territory of Russia.

Endemic Cholera in India and Imported Cholera in Russia: What is Common?

Монахова

Ghosh

Mutreja

et al. 2020

The endemicity of cholera in India has been well researched. Among the other endemic areas, Indian subcontinent appears to be the cradle of Vibrio cholerae genovariants, which subsequently spread worldwide. In contrast, all the cholera cases recorded in Russia are of imported origin. In the past century, such importations might result in epidemics, which, however, ended with elimination of toxigenic V. cholerae (TVC) from the affected areas. Currently, the incidence of TVC in water reservoirs or infected returnees from Asian countries are rare events, mostly due to constant surveillance activities. Furthermore, the climatic conditions in the majority of Russian regions are unfavorable for longterm environmental survival of the pathogen. On the other hand, global shifts in climate accompanied by unexpected anomalies in previously stable climatic zones may promote dissemination of imported TVC and emergence of cholera. In some regions of Russia, seasonal weather patterns are pretty similar to Indian. Molecular studies of isolates from diverse territories show that TVC have been introduced into new areas and acquired additional traits, absent in their ancestors.This article describes genomic characteristics of TVC isolates from Russia, India and some other countries. The review is complemented by bioinformatics analysis of important genetic markers to discriminate between clones that emerged in endemic regions and were imported into cholera-free locations. We have found more evidence in support of primary Indian origin of recent genovariants and their ongoing transformation, including the emergence of a new rtxA gene allele with a 60 bp deletion in addition to the previously known null mutation. In certain conditions, TVC could pose a potential threat of spreading epidemic cholera into Russia and other territories. Cholera control requires joint efforts of researchers to learn more about the pathogen at the molecular level for developing effective strategies to protect the humankind throughout the world.