Biomedical subjects

Waleria Hryniewicz

Publications and source records attributed to Waleria Hryniewicz.

At least 19 recordsLinked to original sources

PubMed2007

Clonal diversity and resistance mechanisms in tetracycline-nonsusceptible Streptococcus pneumoniae isolates in Poland.

The frequency of tetracycline resistance in Streptococcus pneumoniae isolates in Poland is one of the highest in Europe. The aim of this study was to analyze the clonal diversity and resistance determinants of tetracycline-nonsusceptible S. pneumoniae isolates identified in Poland and to investigate the effect of tetracycline resistance on their susceptibilities to tigecycline, doxycycline, and minocycline. We have analyzed 866 pneumococcal isolates collected from 1998 to 2003 from patients with respiratory tract diseases, and 242 of these (27.9%) were found to be resistant to tetracycline. All of the resistant isolates were characterized by testing of their susceptibilities to other antimicrobials, serotyping, pulsed-field gel electrophoresis (PFGE), and identification of tetracycline resistance genes and transposons. Selected isolates representing the main PFGE types were analyzed by multilocus sequence typing. Among the isolates investigated, 27 serotypes and 146 various PFGE patterns, grouped into 90 types, were discerned. The most common PFGE type, corresponding to serotype 19F and sequence type 423, was represented by 22.3% of all of the tetracycline-resistant isolates. The tet(M) gene was the sole resistance gene in the group of isolates studied, and in over 96% of the isolates, the Tn916 family of tet(M)-containing conjugative transposons was detected. Several isolates contained specific variants of the transposons, the Tn1545-like, Tn3872-like, or Tn2009-like element. The correlation between the MICs of tetracycline, doxycycline, and minocycline was revealed, whereas no cross-resistance to tetracycline and tigecycline was observed.

Bacterial Proteins↗

PubMed2006

Clonal structure of Enterococcus faecalis isolated from Polish hospitals: characterization of epidemic clones.

To study the population structure of Enterococcus faecalis from Polish hospitals, 291 isolates were typed by pulsed-field gel electrophoresis and a novel multilocus sequence typing scheme (P. Ruiz-Garbajosa et al., J. Clin. Microbiol. 44:2220-2228, 2006). The isolates originated from geographically widespread medical institutions and were recovered during a 10-year period (1996 to 2005) from different clinical sources. The analysis grouped the isolates into five epidemic and 71 sporadic clones. The importance of the previously identified global clonal complexes CC2 and CC9 was corroborated by our findings that two of the Polish epidemic clones, A and J, were classified into these clonal complexes (CCs). However, the two most predominant clones, C (ST40) and F (CC87), did not cluster in the aforementioned CCs and may represent novel epidemic CCs. These clones may have emerged in Central Europe. Clone F, carrying glycopeptide resistance determinants of VanA or VanB phenotypes, caused several outbreaks in hematology units and appeared to be the most prevalent clone in recent years in Poland. Antimicrobial susceptibility testing and additional tests for pathogenicity-related phenotypes (hemolysin and gelatinase production) and genes (asa1 and esp) were performed to further characterize these epidemic clones. Multidrug resistance, glycopeptide resistance, presence of asa1, and production of hemolysin appeared to be statistically significant features related to epidemicity. Production of gelatinase was significant for two of the epidemic clones, whereas presence of the esp gene was not specific for the epidemic clones.

Anti-Bacterial Agents↗

PubMed2006

Phenotypic and molecular analysis of penicillin-nonsusceptible Streptococcus pneumoniae isolates in Poland.

beta-Lactams are the drugs of choice for the treatment of infections caused by the important bacterial pathogen Streptococcus pneumoniae. The recent growth of resistance of this organism to penicillin observed worldwide is of the highest concern. In this study, using 887 surveillance pneumococcal isolates recovered in Poland from 1998 to 2002, we observed the increase in penicillin nonsusceptibility from 8.7% to 20.3%. All of the 109 penicillin-nonsusceptible S. pneumoniae (PNSP) isolates identified, together with 22 archival PNSP isolates from 1995 to 1997, were subsequently analyzed by susceptibility testing, serotyping, profiling of pbp genes, pulsed-field gel electrophoresis, and multilocus sequence typing (MLST). Four predominant serotypes, serotypes 6B, 9V, 14, and 23F, characterized 85.5% of the isolates. MLST revealed the presence of 34 sequence types, 15 of which were novel types. Representatives of seven multiresistant international clones (Spain(23F)-1, Spain(6B)-2, Spain(9V)-3, Taiwan(23F)-15, Poland(23F)-16, Poland(6B)-20, and Sweden(15A)-25) or their closely related variants comprised the majority of the study isolates. The spread of Spain(9V)-3 and its related clone of serotype 14/ST143 has remarkably contributed to the recent increase in penicillin resistance in pneumococci in the country.

Anti-Bacterial Agents↗

PubMed2006

Characteristics of Staphylococcus aureus strains isolated in Poland in 1996 to 2004 that were deficient in species-specific proteins.

One hundred seventy Staphylococcus aureus isolates, collected in 1996 to 2004, were reidentified by phenotypic and genotypic methods. One hundred ten of these (65%) were confirmed, as previously denoted, to be clumping factor (CF)- or free coagulase-deficient S. aureus, based on their phenotype. Based on the CF or coagulase production, three groups of phenotypically deficient S. aureus isolates were distinguished. Group 1 encompassed CF-positive and coagulase-deficient isolates, group 2 consisted of CF-deficient and coagulase-positive isolates, and group 3 included isolates that were CF positive, had delayed coagulase activity, and were deficient in other species-specific features. All investigated strains harbored the clfA, clfB, coa, spa, and nuc genes, but the presence of their products was not detected by the phenotypic methods. Glycopeptide susceptibility testing showed that 26 isolates (23.6%) were hetero-glycopeptide-intermediate S. aureus(hGISA) or hetero-teicoplanin-intermediate S. aureus (hTISA), based on the population analysis profile. The relatedness of the isolates was evaluated by multiple-locus variable number of tandem repeats analysis, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing. The phenotypically deficient S. aureus isolates were classified into PFGE types B (ST239-III) and D (ST246-IA) and were related to the common clones, Hungarian and Iberian, respectively, which have been widely disseminated in Poland and globally. The simultaneous occurrence of hGISA/hTISA and the CF-deficient phenotypes was found for 62.1% of isolates belonging to group 2. The majority of these isolates were assigned to the Iberian clone (PFGE type D; ST247-IA). An association between the defect in coagulase and that in thermonuclease production was observed, which concerned 59.2% of isolates of group 1. The majority of these isolates belonged to the Hungarian clone (PFGE type B; ST239-III).

Bacterial Typing Techniques↗

PubMed2006

Group A streptococci from invasive-disease episodes in Poland are remarkably divergent at the molecular level.

Forty-one clinical isolates of group A streptococcus (GAS) were recovered in Poland from patients with severe invasive infections and were analyzed by phenotypic and genotypic techniques. All isolates were characterized by determining their susceptibilities to antimicrobial agents and by determining their types by pulsed-field gel electrophoresis, multilocus sequence typing, emm typing, and the detection of five streptococcal pyrogenic exotoxin genes (speA, speB, speC, speF, ssa). The isolates studied were fully susceptible to penicillin G, levofloxacin, quinupristin-dalfopristin, and linezolid. Resistance to tetracycline, chloramphenicol, and erythromycin was detected in 46.3, 12.1, and 9.8% of the isolates, respectively. A total of 23 different emm sequence types were identified, of which emm1 and emm12 (19.5% each) were the most common, followed by emm81, emm44/61, and emm85. All the emm1 isolates had the speA2 allele. Twenty-three unrelated sequence types (STs) were identified, with the most frequent STs, ST28 and ST36, corresponding to emm1 and emm12, respectively. Six newly found STs (STs 375, 376, 377, 378, 379, and 385) corresponded to emm types 74, 102, 77, 76, 84 and 63, respectively. The emm1 type and the presence of speA2 gene were associated with the severity of GAS infections. This work presents the first molecular study on Polish invasive GAS isolates.

Adolescent↗

PubMed2006

Invasive meningococcal disease associated with a very high case fatality rate in the North-West of Poland.

The aim of the study was to investigate invasive meningococcal disease in the North-West of Poland, associated with a case fatality rate of 42.9%, where among the first 11 cases, eight had fatal outcome. All fatal cases were diagnosed as fulminant meningococcal severe sepsis with Waterhouse-Friderichsen syndrome and disseminated intravascular coagulation. Serotyping and pulsed-field gel electrophoresis analysis revealed that the high case fatality rate was not associated with the dissemination of one epidemic clone. However, six cases, all with good outcomes, were caused by C:2b:(P1.2,P1.5) isolates of the same pulsed-field gel electrophoresis type belonging to ST8 complex/Cluster A4.

Adolescent↗

PubMed2006

Molecular epidemiology of acquired-metallo-beta-lactamase-producing bacteria in Poland.

We have analyzed 40 metallo-beta-lactamase (MBL)-producing isolates of Pseudomonas aeruginosa (n = 38), Pseudomonas putida (n = 1), and Acinetobacter genospecies 3 (n = 1) from 17 hospitals in 12 cities in Poland that were identified in 2000 to 2004. Pulsed field gel electrophoresis typing classified the P. aeruginosa isolates into eight types, with two types differentiated further into subtypes. Each of the types was specific either to a given center or to several hospitals of the same or neighboring geographic area. Almost all of the organisms produced beta-lactamase VIM-2; the only exceptions were several P. aeruginosa isolates from two centers which expressed VIM-4. The bla(VIM) genes resided exclusively within class 1 integrons, and these were located in either chromosomal or plasmid DNA. PCR-restriction fragment length polymorphism study of the variable regions of the integrons, followed by DNA sequencing, revealed the presence of eight different, mostly novel gene cassette arrays, six of which contained bla(VIM-2) and two of which contained bla(VIM-4). The occurrence of the integron variants correlated well with the geographic distribution of the MBL-producing organisms, and this suggested that their emergence in particular parts of the country had been likely due to a number of independent events. The following regional dissemination of MBL producers could be attributed to various phenomena, including their clonal spread, horizontal transmission of resistance determinants, or both. All of the data collected in this study revealed that even at this early stage of detection, the epidemiological situation concerning MBL producers in Poland has already been complex and very dynamic.

Acinetobacter↗

PubMed2006

Multilocus sequence types, serotypes, and variants of the surface antigen PspA in Streptococcus pneumoniae isolates from meningitis patients in Poland.

Meningitis caused by Streptococcus pneumoniae represents an important factor of morbidity and mortality in humans. In a significant number of cases, this disease is associated with specific clones of the organism, the so-called invasive pneumococcal clones. The aim of the study was to analyze 156 S. pneumoniae isolates identified as etiological agents of meningitis in Poland in the years 1997 through 2002. The isolates were characterized by multilocus sequence typing (MLST), and the results were compared with those obtained by pulsed-field gel electrophoresis (PFGE) and with the MLST data on invasive pneumococci from other countries. Eighty-nine different sequence types were found in the group of isolates, 50 of which had been known before including 19 of the major invasive clones. However, a significant fraction of the isolates possessed novel combinations of known and new MLST alleles. The majority of penicillin-nonsusceptible isolates belonged to the group of international multiresistant clones (Spain(23F)-1, Spain(6B)-2, Spain(9V)-3, Poland(23F)-16, and Poland(6B)-20), which underlined the importance of these in the dissemination of antimicrobial resistance. The results of the MLST analysis correlated well with the PFGE data, thus again demonstrating good congruence between the two typing methods for S. pneumoniae. The majority of the isolates (95.5%) belonged to families 1 or 2 of the surface protein PspA, confirming its potential usefulness as the vaccine antigen candidate.

Animals↗

PubMed2006

Comparison of PCR-based methods for typing Staphylococcus aureus isolates.

In this study, we compared the potentials of (i) a multiplex PCR-based multilocus variable-number tandem repeat (VNTR) assay; (ii) a triplex PCR coamplifying fragments of spa, coa, and the hypervariable region adjacent to the mecA gene; (iii) restriction profile analysis of the STAR repetitive element; (iv) randomly amplified polymorphic DNA analysis; (v) inter-IS256 PCR; and (vi) rep-MP3 PCR. Multilocus VNTR typing and triplex PCR (coa, spa, and hypervariable region) approaches showed excellent reproducibility and high discriminatory power; however, only multilocus VNTR typing could distinguish all pulsed-field gel electrophoresis and spa types. Multilocus VNTR typing appears to be the most useful PCR-based method for the rapid genotyping of Staphylococcus aureus strains.

Bacterial Typing Techniques↗

PubMed2006

Distribution of the serine-aspartate repeat protein-encoding sdr genes among nasal-carriage and invasive Staphylococcus aureus strains.

The sdr locus was found in all 497 investigated Staphylococcus aureus strains, although in 29 strains it contained only the sdrC gene (sdrD negative, sdrE negative). The sdrC-positive, sdrD-negative, sdrE-negative gene profile was exclusive to methicillin-sensitive S. aureus (MSSA) strains (Fisher's exact test; P = 0.0005) and was not found in the strains collected from bone infections (P = 0.0019). We also found a strong association between the presence of the sdrD gene and methicillin-resistant S. aureus strains (P < 0.0001). Our findings suggest that MSSA strains with the newly uncovered sdrC-positive, sdrD-negative, sdrE-negative gene profile have a substantially decreased potential to establish bone infection.

Bacterial Proteins↗

PubMed2006

[Self-medication with antibiotics in Poland].

UNLABELLED: Antibiotic resistance, the important public health threat, depends on antibiotic overuse/misuse. Self-medication with antibiotics is of serious medical concern. The aim of the study, as a part of SAR project (Self-medication with antibiotic in Europe) was to survey the incidence of this phenomenon. MATERIAL AND METHODS: In Poland among 938 respondents of the questionnaire 27% adults were treated with antibiotics at least once a year, 13% of the therapies were undertaken without medical consultation. The most frequently prescribed antibiotic was amoxicillin, the most often self-ordered--fusafungine. RESULTS: The main self-medicated symptoms were sore throat and common cold. The duration of self-ordered therapy was less then 5 days. The sources of antibiotics used for self-medication were leftovers and those given by friends or family. CONCLUSION: The results of the study confirmed that antibiotic self-medication is relatively frequent problem in Poland and interventions are required in order to reduce the frequency of antibiotics misuse.

Adult↗

PubMed2005

Clonal structure of the methicillin-resistant Staphylococcus aureus (MRSA) population in Poland: revision and update.

The clonal structure of the methicilin-resistant Staphylococcus aureus (MRSA) population in Poland has been analyzed in several reports since the mid-1990s. The present study was performed on 253 MRSA isolates (146 archival and 107 new isolates) recovered in 26 hospitals between 1990 and 2001. Whereas all isolates were typed by pulsed-field gel electrophoresis (PFGE) and the analysis of the ClaI::mecA and ClaI::Tn554 RFLP polymorphism, selected isolates were also subjected to multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) comparisons. Based on the PFGE data, 15 MRSA clones were discerned, seven of which were observed in multiple hospitals. Five of these were related to the pandemic Hungarian (MLST clonal complex, CC8), Iberian (CC8), Pediatric (CC5), Mexican (CC30), and Brazilian clones (CC8). MLST confirmed the earlier reports on the similarity of the Hungarian and Brazilian clones, and it revealed that one of the two remaining epidemic clones was related to the Hungarian/Brazilian, and the other--to the Berlin clones. A local strain from the Northeastern part of the country was found to be similar to a minor Greek clone. The MRSA clonal structure and the increasing complexity of the relationships between the genetic and phenotypic traits of this micro-organism in Poland has now been firmly established.

Chromosomes, Bacterial↗

PubMed2005

Evolution of TEM-type extended-spectrum beta-lactamases in clinical Enterobacteriaceae strains in Poland.

Seventeen extended-spectrum beta-lactamase (ESBL)-producing isolates of the family Enterobacteriaceae recovered from 1998 to 2000 in hospitals of five different cities in Poland were analyzed. They expressed several TEM-type ESBLs, TEM-4, TEM-29, TEM-85, TEM-86, TEM-93, and TEM-94. TEM-85 (L21F, R164S, E240K, T265M), TEM-86 (L21F, R164S, A237T, E240K, T265M), TEM-93 (M182T, G238S, E240K), and TEM-94 (L21F, E104K, M182T, G238S, T265M) were identified for the first time. Including the enzymes described earlier, TEM-47, TEM-48, TEM-49, and TEM-68, the group of known ESBLs of the TEM family produced by enterobacteria in Polish hospitals has increased to 10 variants. Comparative sequence analysis of the genes coding for all these beta-lactamases revealed a view of their possible evolution, which, apart from the gradual acquisition of various mutations, could also have involved recombination events. Two different bla(TEM-1) gene alleles were precursors of the ESBL genes: bla(TEM-1A), which was the ancestor of bla(TEM-93), and bla(TEM-1F), from which all the remaining genes originated. The evolution of the bla(TEM-1F)-related genes most probably consisted of three major separate lineages, one of which, including bla(TEM-4), bla(TEM-47), bla(TEM-48), bla(TEM-49), bla(TEM-68), and bla(TEM-94), was highly structured itself and could have been initiated by the bla(TEM-25) gene, identified exclusively in France so far. Plasmid fingerprinting analysis revealed a high degree of diversity of plasmids carrying related bla(TEM) genes, which suggested either the intense diversification or transposition of bla(TEM) genes between different plasmids or some contribution of convergent evolution. The results of this study clearly demonstrate that the environment of Polish hospitals has been highly favorable for the rapid evolution of ESBLs.

Cloning, Molecular↗

PubMed2005

High genetic diversity of ciprofloxacin-nonsusceptible isolates of Streptococcus pneumoniae in Poland.

We have analyzed the susceptibility to ciprofloxacin of 697 pneumococcal isolates collected in 1998-2002 in Poland from patients with respiratory tract diseases. Thirty-one ciprofloxacin-nonsusceptible isolates (MICs, > or =4 microg/ml) were identified, of which two were resistant to levofloxacin (MIC, 8 microg/ml). Serotyping, pulsed-field gel electrophoresis, multilocus sequence typing, and the analysis of resistance determinants showed their great genetic diversity.

Anti-Infective Agents↗

PubMed2005

Characteristics of Haemophilus influenzae type b responsible for meningitis in Poland from 1997 to 2004.

Two hundred forty-five H. influenzae isolates responsible for meningitis in Poland from 1997 to 2004 were studied. Among these, 233 (95.1%) belonged to serotype b (Hib), 2 belonged to serotype f, and 10 were noncapsulated. The relatedness of all isolates was evaluated by pulsed-field gel electrophoresis (PFGE), and selected representatives were evaluated by multilocus sequence typing. Resistance to ampicillin was identified in 34 (14.6%) of the Hib isolates and was associated with the production of beta-lactamase only. Except for four isolates nonsusceptible to chloramphenicol, all isolates were susceptible to cefotaxime, ciprofloxacin, and rifampin. The PFGE analysis divided the Hib isolates into five PFGE types; however, all of them were possibly related. The most common PFGE type, with 25 subtypes, was characteristic for 97.4% of the isolates. The most prevalent PFGE subtype found in our study was also the most common among the Hib isolates responsible for invasive disease in Italy and the Czech Republic and was found among isolates causing lower respiratory tract infections in Poland. The most prevalent sequence types (STs) in the studied group were ST6 and ST92. Four new STs were found: ST188, ST189, ST190, and ST268. Results of this study support the evidence that the genetic structure of encapsulated H. influenzae is clonal. The continuing high number of meningitis cases due to Hib in Poland underlines the need for mass vaccination against Hib in Poland.

Ampicillin↗

PubMed2005

Prevalence and serotype distribution of encapsulated Haemophilus influenzae isolates from patients with lower respiratory tract infections in Poland.

Among 1,272 H. influenzae isolates obtained from patients with lower respiratory tract infections, 5.6% were found to be encapsulated. The most common were serotypes b (40.3%) and e (38.9%) followed by serotypes f (16.7%) and d (4.1%). All encapsulated isolates within a particular serotype showed high genetic homogeneity and did not produce beta-lactamases.

Adolescent↗

PubMed2005

Comparison of multiple-locus variable-number tandem-repeat analysis with pulsed-field gel electrophoresis, spa typing, and multilocus sequence typing for clonal characterization of Staphylococcus aureus isolates.

Multiple-locus variable-number tandem-repeat analysis (MLVA), a new PCR-based method of typing Staphylococcus aureus, was compared to pulsed-field gel electrophoresis (PFGE), spa typing, and multilocus sequence typing (MLST) on a group of 59 S. aureus (mostly methicillin-resistant) clinical isolates. The aim of the study was to establish possible criteria of clustering MLVA patterns and to check concordance levels between the results produced by MLVA and the three other typing methods. As in our earlier study, MLVA turned out to have discriminatory power similar to that of PFGE. Comparison of data obtained by the two approaches allowed us to propose a 70% or ca. 80% cutoff value of the similarity between two MLVA patterns, depending on a cutoff level applied to interpret the PFGE results, 75% or ca. 90%, respectively. The cutoff values corresponded to the difference of up to six or four bands, respectively, among maximum 14 bands in total produced by two isolates in the analysis. The MLVA clusters matched well those obtained by PFGE, and they were also consistent in general with clusters generated by spa typing and MLST, these latter methods characterized lower resolution. Our results suggest that MLVA may be reliable in shorter-term S. aureus epidemiological studies, including analyses of outbreaks and hospital-to-hospital strain transmission events. Well-known advantages of typing methods based on PCR (low cost, short time, and easiness of performance) make MLVA a method that may be useful in a variety of laboratories, including those performing routine microbiological analyses within medical centers.

Anti-Bacterial Agents↗

PubMed2005

Lysostaphin as a potential therapeutic agent for staphylococcal biofilm eradication.

The aim was to study the activity of lysostaphin in monotherapy or in combination with oxacillin, towards biofilms built by clinical and reference S. aureus and S. epidermidis strains in the wells of microplate, in the chambers of a LabTekII chamber slide or on the polyethylene catheter. MICs of oxacillin and lysostaphin for planktonic bacteria were determined according to the standards of NCCLS. BIC (Biofilm Inhibitory Concentration) was estimated by the MTT assay. The integrity of biofilm treated with antimicrobials was also examined: by staining with FITC and laser scanning fluorescence confocal microscopy and visually by TTC reduction assay. Despite the fact that susceptibility of planktonic cultures of 25 staphylococcal strains to lysostaphin action was various, we have demonstrated the effectiveness of lysostaphin in the treatment of biofilm, built not only on the flat surface of the microplates but also on catheter's surface. The synergistic effect of subBIC lysostaphin+oxacillin was observed for MSSA and MRSA biofilms but not for 1474/01 hVISA strain. Also BICOXA for S. epidermidis RP12 and A4c strains, but not for 6756/99 MRSE biofilms was reduced when lysostaphin was simultaneously used.

Anti-Infective Agents, Local↗