scholarly journals RETROSPECTIVE ANALYSIS OF BIOLOGICAL AND MOLECULAR-GENETIC PROPERTIES OF STRAINS - CAUSATIVE AGENTS OF CHOLERA - ISOLATED IN UKRAINE IN 1994 - 2011

Author(s):  
I. V. Savelieva ◽  
S. N. Tikhonov ◽  
V. N. Saveliev ◽  
D. A. Kovalev ◽  
S. V. Pisarenko ◽  
...  

Aim. Retrospective analysis of biological and molecular-genetic properties of strains - causative agents of cholera - isolated in the period of epidemics in Ukraine in 1994 - 2011. Materials and methods. Phenotypic and molecular-genetic properties of 5 strains of cholera vibrios, biovar El Tor isolated from cholera patients and 4 strains from the environmental samples were studied using traditional bacteriological and genetic methods. Detection of DNA for toxigenicity genes and genes characteristic for El Tor and classic biovar were carried out by PCR method using reagent kits «AmpliSens- Vibrio cholerae FRT» and «.Vibrio cholerae ctxB-rstR-rstC genes, REF» (an experimental test system). Sequencing of genomes of 4 strains of causative agents of cholera was carried out in genetic analyzer Ion Torrent Personal Genome Machine. Results. Strains of cholera vibrios identified in Ukraine in 1994 and 2011 such as a typical toxigenic biovar El Tor (V cholerae 01, El Tor, Ogawa, Hly-, ctxA+, tcpA+) contain genes of the classic cholera vibrio in their genome and are genetically altered (hybrid) variants of cholera vibrio biovar El Tor producing enterotoxin CT1 and having increased virulence, that was clinically manifested in predominance of severe forms of cholera in Mariupol of Donetsk region in 2011. Genome sequences of the 4 studied strains were deposited into the international database DDBJ/EMBL/GenBank. Conclusion. The studied isolates were established to belong to a clade of strains associated with cholera outbreaks in Haiti and Asian continent, from where genetically altered strains of cholera vibrios biovar El Tor were introduced to Haiti in 2010, based on results of comparison of genomic sequences of the studied strains with genomes of V. cholera strains from the international database GenBank.

Author(s):  
I. V. Savelieva ◽  
A. N. Kulichenko ◽  
V. N. Saveliev ◽  
D. A. Kovalev ◽  
O. V. Vasilieva ◽  
...  

Aim. Conduct in a comparative aspect MLVA-typing of genetically altered cholera vibrio biovar El Tor, isolated from patients during the epidemic (1994) and outbreaks (1993, 1998) in Dagestan with isolates in Mariupol (Ukraine) in 1994-2011 in Moscow (2010, 2012), India (1964, 2006, 2007), Bangladesh 1991, 1994, 2001, 2004) and to establish Phylogenetic connections between strains of cholera vibrios isolated in different years in these territories, to ascertain the source of their drift. Materials and methods. MLVA-tyP-ing was carried out in PCR at 5 variable loci of 35 clinical strains of genetically modified Vibrio cholerae byotyPe El Tor. The obtained amPlicon was studied in the system of automatic caPillary electroPhoresis ExPerion («Bio Rad Laboratories», USA). For Phylogenetic analysis, along with MLVA-genotyPes, 35 strains of Vibrio cholerae from the Institute's collection used Published genotyPes of strains isolated in India, Bangladesh, Haiti. Results. The investigated strains of cholera vibrio are referred to 21 MLVA-tyPes, divided into 2 main clades and 1 seParate branch with clonal clusters and subclusters, each of which contains closely related strains of cholera vibrio genovariants having a different degree of Phylogenetic relationshiP - full or Partial identity of allelic Profiles of five variable loci. The sources of drift of genetically modified Vibrio cholerae byotyPe El Tor to Russia and Ukraine from disadvantaged cholera of India, Bangladesh, Azerbaijan and the countries of the Middle East have been established. Conclusion. The obtained data testify to the PolymorPhism of MLVA-tyPes of genetically altered strains of cholera vibrio of the biologist El Tor, evolved in different years and caused ePidemics or outbreaks of cholera in different territories during different time Periods of the course of the seventh cholera Pandemic, and also suggest the Polyclonal origin of the Vibrio cholerae biovar El Tor and the source of their drift to the territory of the Russian Federation and Ukraine.


2012 ◽  
Vol 27 (2) ◽  
pp. 61-68 ◽  
Author(s):  
L. V. Mironova ◽  
S. V. Balakhonov ◽  
L. Ya. Urbanovich ◽  
A. S. Kozhevnikova ◽  
V. S. Polovinkina ◽  
...  

2013 ◽  
Vol 57 (8) ◽  
pp. 3950-3959 ◽  
Author(s):  
Hongxia Wang ◽  
Li Zhang ◽  
Anisia J. Silva ◽  
Jorge A. Benitez

ABSTRACTVibrio choleraestrains of serogroups O1 and O139, the causative agents of the diarrheal illness cholera, express a single polar flagellum powered by sodium motive force and require motility to colonize and spread along the small intestine. In a previous study, we described a high-throughput assay for screening for small molecules that selectively inhibit bacterial motility and identified a family of quinazoline-2,4-diamino analogs (Q24DAs) that (i) paralyzed the sodium-driven polar flagellum ofVibriosand (ii) diminished cholera toxin secreted by El Tor biotypeV. cholerae. In this study, we provide evidence that a Q24DA paralyzes the polar flagellum by interacting with the motor protein PomB. Inhibition of motility with the Q24DA enhanced the transcription of the cholera toxin genes in both biotypes. We also show that the Q24DA interacts with outer membrane protein OmpU and other porins to induce envelope stress and expression of the extracellular RNA polymerase sigma factor σE. We suggest that Q24DA-induced envelope stress could affect the correct folding, assembly, and secretion of pentameric cholera toxin in El Tor biotypeV. choleraeindependently of its effect on motility.


1940 ◽  
Vol 40 (5) ◽  
pp. 532-537 ◽  
Author(s):  
W. James Wilson ◽  
L. V. Reilly

1. A fluid enrichment medium containing sulphite and bismuth based on Read's modification of one of Wilson & Blair's media has been devised and the claims of Read (1939) and Seal (1939) as to the value of such a medium has been confirmed.2. A saccharose mannitol sulphite bismuth alcohol phenol red agar medium is described and has been found to allow rich growth of the true cholera vibrio and to inhibit the growth not only of B. coli and B. lactis aërogenes but of many vibrios liable to be mistaken for the true vibrio cholerae.3. Thirty-one strains of true cholera vibrios, obtained from the National Collection, grew rapidly and profusely in the media.4. Of twenty-five cholera-like and para-cholera strains six grew well, whilst the growth of nineteen was scanty or nil.5. Of eleven El Tor vibrios five grew in both fluid and solid bismuth media, five grew only on the solid medium and in one no growth occurred in either medium. The El Tor colonies were much smaller than those of the epidemic cholera strains.


Author(s):  
Z. A. Sagiev ◽  
R. S. Musagalieva ◽  
A. A. Abdirasilova ◽  
T. Z. Ayazbaev ◽  
M. M. Kul’baeva ◽  
...  

In 2017, from October 15 to November 21, 5 cholera cases imported from India – 3 patients and 2 carriers of V. cholerae – were recorded in Almaty. The patients recovered from the disease. Objective of the study was to characterize the imported cases of cholera and investigate the properties of cholera vibrio strains isolated from patients and carriers of V. cholerae. Materials and methods. Revised were the medical records; blood sera, feces from patients and contact persons were assayed. Studied were sensitivity spectrum to antibacterial preparations of isolated V. cholerae strains according to the “Methodological guidelines on laboratory diagnosis of cholera”, dated September 27, 2010; No 252. Epidemiological, microbiological, immunological and molecular-genetic methods were applied for investigation. Results and conclusions. Consequently to molecular genetic studies, genes of specificity, wbeN and toxicity (epidemic significance), ctxA, tcpA were detected in samples from 3 patients and 2 contact persons. The isolated strains were identified as Vibrio cholerae O1 Eltor Inaba in two cases, and in one case – as Vibrio cholerae O1 Eltor Hykoshima, Heiberg group I, toxigenic, hemolysis negative in Greig test, virulent, highly sensitive to ciprofloxacin, doxycilin, erythromycin, tetracycline and moderately sensitive to levomycetin. It was established that the country of export in all the cases was India. Relevant anti-epidemic and preventive measures were undertaken to localize and eradicate the foci in order to prevent possible threat of epidemic spread of infections among the population.


Author(s):  
V.A. Korshenko ◽  
I.Ya. Cherepakhina

Anti-lactoferrin activity (AlfA) is one of the factors contributing to the persistence of V. cholerae in human body in formation of human vibriocarriers. It is shown that in the AlfA mechanism cholera vibrio lectins are participating as an initial link specifically binding to lactoferrin at the surface of microbial cells, providing for subsequent cleavage of this protein with the help of V. cholerae exoproteases. Lectins are proteins or glycoproteins of non-immune class, capable to reversibly bind to the carbohydrate fraction of glycoconjugates without breaking the covalent structure of any of the recognizable glycosyl ligands. As glycoproteins, lectins ensure the complementary highly specific carbohydrate-protein binding of the active sites of target cells (namely lactoferrin). It is proved that all V. cholerae El Tor and Bengal strains possess hexose-specific lectin receptors (especially mannose). Atoxigenic V. cholerae O1 and O139 strains possess, in addition to hexose receptors, lectins specific to aminosugars.


2009 ◽  
Vol 58 (2) ◽  
pp. 239-247 ◽  
Author(s):  
Souvik Chatterjee ◽  
Tapas Patra ◽  
Kausik Ghosh ◽  
Amit Raychoudhuri ◽  
Gururaja P. Pazhani ◽  
...  

Retrospective analysis led to the detection of two Vibrio cholerae variant O1 strains (VC51 and VC53), which were isolated in 1992 in Kolkata from clinical cases, with identical traits to 2004 Mozambique variant O1 strains. The Mozambique O1 strains that caused a huge outbreak in 2004 have been shown to have phenotypic traits of both classical and El Tor biotypes, and thereby have been reported as variant. Our study demonstrated that two O1 strains isolated in Kolkata during 1992 were of the El Tor background as evidenced by polymyxin B (50 U ml−1) resistance, positivity in Voges–Proskauer reactions and sensitivity to biotype-specific vibrio phages. With the features of classical CTX prophage, localization in the small chromosome, and an absence of RS1 and pTLC, both Mozambique and Kolkata strains appeared to be identical. Furthermore, two Kolkata strains exhibited an identical ribotype to that of the Mozambique variant, displaying ribotype pattern RI that had been assigned to Kolkata V. cholerae O1 strains isolated on or before 1992. NotI pulsotype analysis indicated that these 1992 Kolkata strains along with the Mozambique variant O1 belonged to very closely related clones. Considering the chronological events, and the typical identity at the phenotypic and the genotypic level between the two O1 strains isolated during 1992 from Kolkata and during 2004 from Mozambique, we propose that some of the 1992 Kolkata O1 strains might have acted as progenitors for Mozambique variant O1 strains.


Author(s):  
O. V. Duvanova ◽  
B. N. Mishankin ◽  
S. V. Titova ◽  
L. A. Korneeva

Aim. To study the effect of N-acetyl-L-cysteine on biofilm of V. cholerae of different sero-groups isolated from various sources and with different epidemiological significance (the presence/ absence of the ctx AB genes and tcpA). Materials and methods. Bacterial eiiltiire of Vibrio cholerae El Tor O1 and O139 serogroups were grown as biolms. We have estimated the influence of the drug N-acetyl-L-cysteine at a concentration of 0.5 - 4 mg/ml on the formation, of the formed biofilm and in planktonic form. Results. Discovered antibacterial activity of N-acetyl-L-cysteine. Noted that it was influenced as in the formation, of the already formed biofilm and the planktonic form, the representatives of the Vibrio cholerae El Tor O1 and O139 serogroups in concentrations of 2 - 4 mg/ml, showing an antibacterial effect regardless of the presence/absence of genes ctx and tcpA AB. Conclusion. Identified the antibacterial action of the drug N-acetyl-L-cysteine against biofilm of V. cholerae indicates the desirability of considering the possibility of using drug therapy in cases variety of diseases caused by causative agents II - IV groups pathogenicity.


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