RESUMO
Were studied the genes encoding the virulence factors of 221 strains: E. coli O6:H1 (194) and E. coli O25:H4 (27), isolated in 2014-2018 from stool samples of children and adults examined according to epidemic indications. Molecular methods included PCR with hybridization-fluorescence and electrophoresis detection of amplified products. The strains did not have virulence genes for diarrheagenic E. coli (DEC) pathogroups EPEC, ETEC, EIEC, EHEC, EAggEC, and belonged to the phylogenetic group B2. They contained from four to eight genes encoding virulence factors of ExPEC: E. coli O6:H1 - pap (68,6%), sfa (87,6%), fimH (96,4%), hly (62,4%), cnf (74,7%), iutA (97,9%), fyuA (95,9%), chu (100%); E. coli O25:H4 - pap (66,7%), afa (22,2%), fimH (100%), hly (44,4%), cnf (44,4%), iutA (100%) , fyuA (100%), chu (100%). The antimicrobial susceptibility testing to 6 classes of antimicrobials (beta-lactams, fluoroquinolones, aminoglycosides, nitrofurantoin, sulfanilamide, trimethoprim / sulfamethoxazole) according the EUCAST. 60,3% of E. coli O6:H1 were sensitive to antibiotics, E. coli O25:H4 remained sensitive to carbapenems and nitrofurans. Extended-spectrum cephalosporins resistance was due to the production ESBL (CTX-M). The 57,1% resistant strains of E. coli O6:H1 and 100% of E. coli O25:H4 strains belonged to the MDR phenotype. The XDR phenotype had one in five MDR strains of E. coli O6:H1 and E. coli O25:H4. All strains of E. coli O25:H4 belonged to ST131. Given the important role of E. coli in human pathology, detection of virulence genes should be performed to confirm the etiological significance of the isolated strain.
Assuntos
Infecções por Escherichia coli/diagnóstico , Escherichia coli/classificação , Sorotipagem , Adulto , Criança , Escherichia coli/efeitos dos fármacos , Genes Bacterianos , Humanos , Testes de Sensibilidade Microbiana , Filogenia , Fatores de Virulência/genética , beta-LactamasesRESUMO
The article presents the results of antimicrobial resistance monitoring of Salmonella isolated from children and adults with diarrhea in St. Petersburg in 2014-2018. In 746 isolates of 42 serovars more than 90,0% belonged to three: S. enteritidis (79,6%), S. typhimurium (6,8%) and S. infantis (3,8%). The antimicrobial susceptibility testing (according the EUCAST) to 7 classes of antimicrobials revealed the resistance in 78,6% of Salmonella. Low-level quinolone resistance (MIC of ciprofloxacin 0,12-0,5 mg/l) was detected in 63,3% isolates (S. enteritidis -71,0%, S. typhimurium - 15,7%, S. infantis - 89,3%) and was due to five kinds of single nucleotide substitutions in gyrA: Asp87Tyr - 36,1% of studied isolates (only S. infantis); Ser83Phe - 22,2% (only S. enteritidis); Asp87Asn - 19,4% (S. enteritidis, S. typhimurium, S. hadar, S. newport); Ser83Tyr -11,1% (S. enteritidis and S. infantis) and Asp87Gly - 8,3% (only S. enteritidis). Only in one S. kentucky isolate with high-level fluoroquinolone resistance (MIC of ciprofloxacin > 8,0 mg/l) two substitutions (Ser83Phe and Asp87Asn) were detected. Two Salmonella isolates (S. typhimurium and S. corvallis) had plasmid-mediated quinolone resistance (qnrS). Extended-spectrum cephalosporin resistance was found in 6 Salmonella serovars (1,6%). The bla-genes were detected: of genetic group CTX-M1 - in 10 isolates (serovars S. typhimurium, S. enteritidis, S. abony, S. coeln and S. virchow), CTX-M2 - in 2 S. typhimurium isolates, CTX-M9 - in three S. enteritidis isolates. In one S. typhimurium CTX-M1 and CTX-M2 were detected. The gene of CMY-2 (molecular class C cephalosporinase) was revealed in two isolates (S. newport and S. enteritidis). Our study showed that Salmonella (the main bacterial pathogen of acute diarrhea in children and adults) isolated in Saint-Petersburg had antimicrobial resistance to drugs of choice for salmonellosis treatment.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Salmonella/efeitos dos fármacos , Adulto , Criança , Humanos , Testes de Sensibilidade Microbiana , Federação RussaRESUMO
UNLABELLED: Shiga toxin-producing Escherichia coli (STEC) food-borne infections are reported worldwide and represent a serious problem for public healthcare. In the Russian Federation there is little information on epidemiology and etiology of STEC-infections as well as on molecular-genetic peculiarities of STEC pathogens. OBJECTIVE: Our aim was to describe a food-borne outbreak as hemorrhagic colitis (HC) along with hemolytic uremic syndrome (HUS), enterocolitis, and acute gastroenteritis in children in St. Petersburg in 2013. METHODS: Epidemiological, microbiological, molecular-genetic and bioinformatic methods were applied. RESULTS: Objects to study were clinical specimens, milk and food samples, as well as STEC strains isolated during the outbreak. The outbreak of food-borne infection was found to be caused by STEC-contaminated raw milk as confirmed by epidemiological analysis, detection of STEC DNA and isolation of relevant pathogens in milk and sick children fecal specimens. The whole-genome sequencing revealed two groups ofpathogens, E. coli O157:H7 and E. coli O101:H33 among collected strains. Group I strains were attributed to the previously known sequence type ST24, while group II strains belonged to the previously non-described sequence type ST145. In strain genomes of both groups there were identified nucleotide sequences of VT2-like prophage carrying stx2c gene, plasmid enterohemolysin gene, and gene of the STEC main adhesion factor intimin. Gene of intimin gamma was identified in E. coli O157:H7 strains and intimin iota 2 in E. coli O101:H33 strains. The latter previously was identified only in enteropathogenic E. coli (EPEC) strains. CONCLUSION: The additional knowledge of epidemiology and biology of STEC pathogens would assist clinicians and epidemiologists in diagnosing, treating and preventing hemorrhagic colitis.
Assuntos
Colite/etiologia , Surtos de Doenças/estatística & dados numéricos , Infecções por Escherichia coli , Síndrome Hemolítico-Urêmica/etiologia , Leite/microbiologia , Escherichia coli Shiga Toxigênica , Animais , Criança , Pré-Escolar , Reservatórios de Doenças , Infecções por Escherichia coli/complicações , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/fisiopatologia , Infecções por Escherichia coli/prevenção & controle , Feminino , Doenças Transmitidas por Alimentos/complicações , Doenças Transmitidas por Alimentos/diagnóstico , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/fisiopatologia , Doenças Transmitidas por Alimentos/prevenção & controle , Estudo de Associação Genômica Ampla , Humanos , Lactente , Masculino , Federação Russa/epidemiologia , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/isolamento & purificaçãoRESUMO
AIM: Determine etiologic significance of clinical strains of E. coli O145:H28 isolated from feces of a patient with hemolytic-uremic syndrome (HUS). MATERIALS AND METHODS: 20 E. coli strains isolated from feces of a patient with HUS that developed after an acute intestine infection were studied. Antigenic structure of strains was determined by sequencing of rib and fliC genes; presence of virulence genes (pap, aaf, sfa, afa, eaeA, bfpA, ial, hly, cnf, stx1, stx2, LT, ST and aer)--in PCR; ESBL production --by double disk method, ESBL genes--in PCR. RESULTS: The strains contained rfb gene coding O145, fliC gene coding H7. Genes coding synthesis of stx2-toxin and intimin (eaeA) were detected. The strains were resistant to beta-lactams due to production of CTX-M class ESBL. CONCLUSION: A causative agent E. coli O145:H28 was isolated from a patient with HUS that produces stx2-toxin and CTX-M class ESBL and has not been previously registered in Russian Federation.
Assuntos
Escherichia coli Êntero-Hemorrágica , Infecções por Escherichia coli , Síndrome Hemolítico-Urêmica , Proteínas de Bactérias/metabolismo , Criança , Escherichia coli Êntero-Hemorrágica/genética , Escherichia coli Êntero-Hemorrágica/isolamento & purificação , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Flagelina , Síndrome Hemolítico-Urêmica/genética , Síndrome Hemolítico-Urêmica/microbiologia , Humanos , Masculino , Federação Russa , Análise de Sequência de DNA , Fatores de Virulência/genéticaRESUMO
The review presents the characteristics of E. coli O104:H4, the causative agent of large-scale alimentary ictus in Germany in spring time 2011. The antigenic characteristics and factors of E. coli pathogenicity are taken into account. The causative agent has a combination of pathogenic factors of two groups of diarrheigenic Escherichia: shigella similar toxin, specific for entero-hemorrhagic E. coli and adhesins of enteroaggregative E. coli.
Assuntos
Antígenos de Bactérias , Surtos de Doenças , Escherichia coli Êntero-Hemorrágica , Infecções por Escherichia coli , Toxina Shiga , Fatores de Virulência , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/metabolismo , Escherichia coli Êntero-Hemorrágica/citologia , Escherichia coli Êntero-Hemorrágica/imunologia , Escherichia coli Êntero-Hemorrágica/metabolismo , Escherichia coli Êntero-Hemorrágica/patogenicidade , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Feminino , Alemanha/epidemiologia , Humanos , Masculino , Toxina Shiga/genética , Toxina Shiga/imunologia , Toxina Shiga/metabolismo , Fatores de Virulência/genética , Fatores de Virulência/imunologia , Fatores de Virulência/metabolismoRESUMO
The surface antigen has been obtained from C. jejuni culture and its immunochemical analysis has been carried out by the methods of immunoelectrophoresis, double diffusion with homologous and heterologous rabbit sera. The protein profile of this antigen has been studied by means of vertical electrophoresis in polyacrylamide gel with sodium dodecyl sulfate, and the molecular weight of the protein components of the antigen has been determined. The presence of anti-Campylobacter antibodies has been revealed in a number of rabbit sera by enzyme immunoassay techniques. The results thus obtained indicate the possibility of developing the enzyme immunoassay for the determination of antibodies in the sera of patients with Campylobacter infection.
Assuntos
Antígenos de Bactérias/análise , Campylobacter jejuni/imunologia , Epitopos/análise , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/isolamento & purificação , Antígenos de Superfície/análise , Antígenos de Superfície/isolamento & purificação , Epitopos/isolamento & purificação , Soros Imunes , Técnicas Imunoenzimáticas , CoelhosRESUMO
Erythrite agar, a commercial culture medium produced by the Research Institute for Culture Media (Makhachkala, USSR), is suitable for the cultivation of C. jejuni after the addition of hemolized sheep blood. The addition of antibiotics to the medium permits its use for the isolation of C. jejuni from feces. By means of this medium C. jejuni has been isolated from the feces of 10% of patients hospitalized on account of acute infectious intestinal diseases.