ABSTRACT
AIM: Study N-acetyl-ß-D-glucosaminidase (chitobiase) (EC 3.2.1.30) in strains of Vibrio cholerae of O1/non-O1 serogroups of various origin, that is a component of chitinolytic complex taking into account object of isolation and epidemiologic significance of strains. MATERIALS AND METHODS: Cultures of V. cholerae O1/non-O1 serogroup strains were obtained from the museum of live culture of Rostov RIPC. Enzymatic activity analysis was carried out in Hitachi F-2500 fluorescent spectrophotometer using FL Solutions licensed software. NCBI databases were used during enzyme characteristics. RESULTS: N-acetyl-ß-D-glucosaminidase in Vcholerae O1/non-O1 serogroup strains was detected, purified by column chromatography, studied and characterized by a number of physical-chemical and biological properties. Comparative computer analysis of amino acid sequence of N-acetyl-ß-D-glucosaminidases of V. cholerae (VC2217 gene), Serratia marcescens etc. has allowed. to attribute the enzyme from V. cholerae to glycosyl-hydrolases (chitobiases) of family 20 and classify it according to enzyme nomenclature as EC 3.2.1.30. CONCLUSION: N-acetyl-ß-D-glucosaminidase in V. cholerae of O1/non-O1 serogroups of various origin and epidemiologic significance, participating in chitin utilization was studied and characterized for the first time, and its possible role in biology of cholera causative agent was shown.
Subject(s)
Acetylglucosaminidase/genetics , Cholera/enzymology , Vibrio cholerae O1/genetics , Vibrio cholerae non-O1/isolation & purification , Acetylglucosaminidase/classification , Amino Acid Sequence , Cholera/epidemiology , Cholera/microbiology , Humans , Serratia marcescens/enzymology , Vibrio cholerae O1/isolation & purification , Vibrio cholerae O1/pathogenicity , Vibrio cholerae non-O1/genetics , Vibrio cholerae non-O1/pathogenicityABSTRACT
AIM: Comparative study of antibiotics resistance and VNTR-typing of Vibrio cholerae non O1/ non O139 strains, isolated on the territory of Rostov region in 2014. MATERIALS AND METHODS: Antibioticogramms of strains were determined by serial dilution method in dense nutrient medium according to MG 4.2.2495-09 (2009). Pheno-, sero- and VNTR-typing was carried out by conventional-methods. RESULTS: The studied strains belonged to V. cholerae species, did not agglutinate with O1 and O139 sera, were atoxigenic hemolysis-positive, did not contain genes of cholera toxin and toxin-coregulating pili of adhesion, contained genes of hemagglutinin/protease, protease PrtV, collagenase, cytotonic factor Cef, outer membrane protein-OmpW, tol- and -vps-clusters, regulatory genes toxR and hapR. Antibioticogramms of the strains have shown the presence of cultures, resistant to ampicillin, ceftazidime-furazolidone, trimethoprim/sulfamethoxazole with intermediate resistance to streptomycin, kanamycin, gentamycin, amikacin, netilmicin, Approximately 20% of isolates had multiple drug resistance. Data of VNTR- and genotyping confirmed a possibility of water transmission route of the infection. CONCLUSION: Execution of monitoring of cultures from environmental samples is necessary for timely detection of genetic characteristics, antibiotics resistance.
Subject(s)
Cholera/epidemiology , Genes, Bacterial , Vibrio cholerae O139/genetics , Vibrio cholerae non-O1/genetics , Water Microbiology , Anti-Bacterial Agents/pharmacology , Bacterial Outer Membrane Proteins/genetics , Bacterial Outer Membrane Proteins/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cholera/drug therapy , Cholera/microbiology , Cholera/transmission , Cholera Toxin/genetics , Cholera Toxin/metabolism , Collagenases/genetics , Collagenases/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Drug Resistance, Multiple, Bacterial , Epidemiological Monitoring , Fimbriae, Bacterial , Gene Deletion , Humans , Immune Sera/chemistry , Metalloendopeptidases/genetics , Metalloendopeptidases/metabolism , Microbial Sensitivity Tests , Peptide Hydrolases/genetics , Peptide Hydrolases/metabolism , Phylogeny , Russia/epidemiology , Serotyping , Transcription Factors/genetics , Transcription Factors/metabolism , Vibrio cholerae O139/classification , Vibrio cholerae O139/drug effects , Vibrio cholerae O139/isolation & purification , Vibrio cholerae non-O1/classification , Vibrio cholerae non-O1/drug effects , Vibrio cholerae non-O1/isolation & purificationABSTRACT
AIM: Application of the authors' GIS <