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Increasing Resistance and Changes in Distribution of Serotypes of Streptococcus agalactiae in Poland.

ABSTRACT: Streptococcus agalactiae is responsible for serious infections in newborn babies, pregnant women, and other patients. The aim of this study was to evaluate antimicrobial susceptibility, serotype distribution, and virulence determinants of the S. agalactiae isolates derived from clinical specimens considering the global increase of both antibiotic resistance and virulence. A total of 165 isolates were identified and serotyped by PCR techniques. Antimicrobial susceptibility was assessed by disk diffusion method, gradient diffusion method and VITEK® System. Virulence associated genes were investigated by PCR; ability to form biofilm was assessed using a microtiter plate assay. The highest observed MIC value for penicillin G was 0.12 µg/mL, seen in 8.5% of isolates. Resistance to erythromycin and clindamycin were found in 30.38% and 24.8% of the strains, respectively. The serotype III (32.73%), V (25.45%), and Ia (18.18%) were found as the most frequently represented. Previously unidentified strains in Poland, belonging to serotypes VI (three strains) and VII (one strain) were recognized. The presence of genes encoding various virulence factors as well as diverse ability to form biofilm were found. In conclusion, macrolide-resistance and decreased susceptibility to penicillin G were revealed signifying the increasing resistance among group B streptococci. Moreover, the presence of genes encoding various virulence factors and the ability to form biofilm were confirmed indicating their role in the pathomechanisms of the evaluated GBS infections.

SUBMITTER: Kaminska D

PROVIDER: S-EPMC7400139 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Increasing Resistance and Changes in Distribution of Serotypes of Streptococcus agalactiae in Poland.

Kaminska Dorota D Ratajczak Magdalena M Szumała-Kąkol Anna A Dlugaszewska Jolanta J Nowak-Malczewska Dorota M DM Gajecka Marzena M

Pathogens (Basel, Switzerland) 20200629 7

Streptococcus agalactiae is responsible for serious infections in newborn babies, pregnant women, and other patients. The aim of this study was to evaluate antimicrobial susceptibility, serotype distribution, and virulence determinants of the S. agalactiae isolates derived from clinical specimens considering the global increase of both antibiotic resistance and virulence. A total of 165 isolates were identified and serotyped by PCR techniques. Antimicrobial susceptibility was asses ...[more]

PMID: 32610654

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Project description:Group B streptococcus (GBS) or Streptococcus agalactiae is an opportunistic pathogen that causes serious illness in newborns, pregnant women, and adults. However, insufficient detection methods and disease prevention programs have contributed to an increase in the incidence and fatality rates associated with this pathogen in non-neonatal patients. This study aimed to investigate factors of the observed increased incidence by investigation of serotype distribution, virulence factors, and antimicrobial susceptibility patterns from invasive GBS disease among non-neonatal patients in Thailand. During 2017-2018, a total of 109 S. agalactiae isolates were collected from non-pregnant patients. There were 62 males and 47 females, with an average age of 63.5 years (range: 20 - 96). Serotypes were determined by latex agglutination assay and multiplex polymerase chain reaction (PCR)-based assay. Among those isolates, seven virulence genes (rib, bca, pavA, lmb, scpB, cylE, and cfb) were detected by PCR amplification, and were determined for their susceptibility to 20 antimicrobial agents using a SensititreTM Streptococcus species STP6F AST plate. Among the study isolates, serotype III was predominant (52.3%), followed by serotype V and serotype VI (13.8% for each), serotype Ib (11.9%), and other serotypes (8.2%). Of the seven virulence genes, pavA was found in 67.0%. Except for one, there were no significant differences in virulence genes between serotype III and non-serotype III. Study isolates showed an overall rate of non-susceptibility to penicillin, the first-line antibiotic, of only 0.9%, whereas the resistance rates measured in tetracycline, clindamycin, azithromycin, and erythromycin were 41.3, 22.0, 22.0, and 22.0%, respectively. Strains that were resistant to all four of those drugs were significantly associated with non-serotype III (p < 0.001). Using multi-locus sequence typing (MLST), 40.0% of the four-drug-resistant isolates belonged to serotype VI/ST1, followed by serotype Ib/ST1 (35.0%). Cluster analysis with global GBS isolates suggested that the multiple drug-resistant isolates to be strongly associated with the clonal complex (CC) 1 (p < 0.001). Compared to the 2014 study of 210 invasive GBS isolates conducted in 12 tertiary hospitals in Thailand, the proportion of serotype III has dramatically dropped from nearly 90% to about 50%. This suggests that resistances to the second-line antibiotics for GBS might be the selective pressure causing the high prevalence of non-serotype III isolates.

Project description:The mreA gene from Streptococcus agalactiae COH31 gamma/delta, resistant to macrolides and clindamycin by active efflux, has recently been cloned in Escherichia coli, where it was reported to confer macrolide resistance (J. Clancy, F. Dib-Hajj, J. W. Petitpas, and W. Yuan, Antimicrob. Agents Chemother. 41:2719--2723, 1997). Cumulative data suggested that the mreA gene was located on the chromosome of S. agalactiae COH31 gamma/delta. Analysis of the deduced amino acid sequence of mreA revealed significant homology with several bifunctional flavokinases/(flavin adenine dinucleotide (FAD) synthetases, which convert riboflavin to flavin mononucleotide (FMN) and FMN to FAD, respectively. High-performance liquid chromatography experiments showed that the mreA gene product had a monofunctional flavokinase activity, similar to that of RibR from Bacillus subtilis. Sequences identical to those of the mreA gene and of a 121-bp upstream region containing a putative promoter were detected in strains of S. agalactiae UCN4, UCN5, and UCN6 susceptible to macrolides. mreA and its allele from S. agalactiae UCN4 were cloned on the shuttle vector pAT28. Both constructs were introduced into E. coli, where they conferred a similar two- to fourfold increase in the MICs of erythromycin, spiramycin, and clindamycin. The MICs of a variety of other molecules, including crystal violet, acriflavin, sodium dodecyl sulfate, and antibiotics, such as certain cephalosporins, chloramphenicol, doxycycline, nalidixic acid, novobiocin, and rifampin, were also increased. In contrast, resistance to these compounds was not detected when the constructs were introduced into E. faecalis JH2-2. In conclusion, the mreA gene was probably resident in S. agalactiae and may encode a metabolic function. We could not provide any evidence that it was responsible for macrolide resistance in S. agalactiae COH31 gamma/delta; broad-spectrum resistance conferred by the gene in E. coli could involve multidrug efflux pumps by a mechanism that remains to be elucidated.

Project description:BackgroundIn pregnant women Streptococcus agalactiae (GBS) can be transmitted to newborn causing severe infections. It is classified into 10 serotypes (Ia, Ib, II-IX). The severity of neonatal disease is determined by the capsular serotype and virulence factors such as the polysaccharide capsule, encoded by the cps gene, protein C, which includes the Cα surface proteins (bca gene), Rib (rib gene) and Cβ (bac gene); the proteins Lmb (lmb gene), FbsB (fbsB gene), FbsA (fbsA gene), the cyl operon encoding a β-hemolysin (hylB gene), the CAMP factor (cfb gene) and the C5a peptidase (scpB gene). The aim of this work was to determine the degree of GBS colonization in pregnant women, the serotypes distribution and to investigate virulence-associated genes.MethodsWe worked with 3480 samples of vagino-rectal swabs of women with 35-37 weeks of gestation. The identification of the strains was carried out using conventional biochemical tests and group confirmatory serology using a commercial latex particle agglutination kit. Two hundred GBS strains were selected. Their serotype was determined by agglutination tests. The monoplex PCR technique was used to investigate nine virulence-associated genes (cps, bca, rib, bac, lmb, fbsB, fbsA, hylB and scpB).ResultsThe maternal colonization was 9.09%. The serotypes found were: Ia (33.50%), III (19.00%), Ib (15.50%), II (14.00%), V (7.00%) and IX (5.50%). 5.50% of strains were found to be non-serotypeable (NT). The nine virulence genes investigated were detected simultaneously in 36.50% of the strains. The genes that were most frequently detected were scpB (100.00%), fbsA (100.00%), fbsB (100.00%), cylB (95.00%), lmb (94.00%) and bca (87.50%). We found associations between serotype and genes bac (p = 0.003), cylB (p = 0.02), rib (p = 0.01) and lmb (p < 0.001).ConclusionsThe frequency of vaginal-rectal colonization, serotypes distribution and associated virulence genes, varies widely among geographical areas. Therefore, epidemiological surveillance is necessary to provide data to guide decision-making and planning of prevention and control strategies.

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Increasing Resistance and Changes in Distribution of Serotypes of Streptococcus agalactiae in Poland.

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Increasing Resistance and Changes in Distribution of Serotypes of <i>Streptococcus agalactiae</i> in Poland.

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