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1.
Eur J Clin Microbiol Infect Dis ; 42(12): 1449-1457, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37857919

ABSTRACT

PURPOSE: To elucidate the role of the Klebsiella oxytoca species complex (KoSC) in epidemiology of VIM-type MBL-producing Enterobacterales in Poland. METHODS: The study comprised all 106 VIM-positive KoSC isolates collected by the Polish National Reference Centre for Susceptibility Testing during 2009-2019 from 60 institutions in 35 towns. All isolates were sequenced by Illumina MiSeq, followed by MinION sequencing of selected organisms. Genomes were subjected to bioinformatic analysis, addressing taxonomy, clonality, phylogeny and structural characterisation of key resistance determinants within their chromosomal and plasmidic loci. RESULTS: Among five species identified, K. oxytoca was predominant (n = 92), followed by Klebsiella michiganensis (n = 11). MLST distinguished 18 STs, with the most prevalent Klebsiella oxytoca ST145 (n = 83). The clone segregated a lineage with the In237-like integron [blaVIM-1-aacA4 genes; n = 78], recorded in 28 cities almost all over the country. The integron was located in a ~ 49-50 kb chromosomal mosaic region with multiple other resistance genes, linked to a ~ 51 kb phage-like element. The organism might have originated from Greece, and its evolution in Poland included several events of chromosomal ~ 54-258 kb deletions, comprising the natural ß-lactamase blaOXY gene. A group of other isolates of various species and clones (n = 12) carried the integron In916 on self-transmissible IncA-type plasmids, effectively spreading in Italy, France and Poland. CONCLUSION: KoSC has been one of the major VIM producers in Poland, owing largely to clonal expansion of the specific K. oxytoca-In237-like lineage. Its apparently enhanced epidemic potential may create a danger on international scale.


Subject(s)
Carbapenem-Resistant Enterobacteriaceae , Klebsiella oxytoca , Humans , Poland/epidemiology , Klebsiella oxytoca/genetics , Multilocus Sequence Typing , beta-Lactamases/genetics , Carbapenem-Resistant Enterobacteriaceae/genetics , Plasmids/genetics , Microbial Sensitivity Tests , Anti-Bacterial Agents/pharmacology , Klebsiella pneumoniae/genetics
2.
Eur J Clin Microbiol Infect Dis ; 42(12): 1523-1530, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37857920

ABSTRACT

One-hundred Polish soldiers of a contingent in Afghanistan in 2019 were screened for Enterobacterales resistant to newer-generation ß-lactams at their departure and return. Seventeen percent were colonized in the gut at the departure, whereas 70% acquired carriage in Afghanistan. The commonest organisms were extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli (ESBL-Ec; 96.6%). All isolates were sequenced and were clonally diverse overall, even within the same sequence type, indicating that independent acquisitions mainly. ESBL-Ec were often multi-drug-resistant. Soldiers stationing in certain regions are at high risk of acquiring resistant bacteria that may cause endogenous infection, be transmitted to vulnerable individuals, and spread resistance genes.


Subject(s)
Escherichia coli Infections , Military Personnel , Humans , Escherichia coli Infections/epidemiology , Escherichia coli Infections/microbiology , beta-Lactamases/genetics , Escherichia coli/genetics , Afghanistan/epidemiology , Anti-Bacterial Agents/pharmacology
3.
J Antimicrob Chemother ; 77(6): 1561-1569, 2022 05 29.
Article in English | MEDLINE | ID: mdl-35296887

ABSTRACT

BACKGROUND: Consecutive Polish regions have become endemic for NDM-1-producing Klebsiella pneumoniae ST11, followed by K. pneumoniae ST147. Since 2017 a significant increase in NDM-positive Enterobacter hormaechei cases has been observed. OBJECTIVES: To investigate the origin and character of this increase in NDM-positive E. hormaechei. METHODS: The analysis included 160 NDM-producing Enterobacter cloacae complex isolates, recovered in 2015-20 in 37 centres of 9/16 regions. These were typed by PFGE and MLST, and screened by PCR-mapping for NDM-1-encoding Tn125-like elements. Forty-four isolates were sequenced by MiSeq. Species identification was based on whole-genome average nucleotide identity; clonality and phylogeny were inferred by SNP approaches. The structural plasmid analysis was done for 12 isolates sequenced by MinION. RESULTS: The isolates belonged to 11 STs, predominantly ST89 (65.6%), followed by ST146 (15.6%), ST198 (7.5%) and ST1303 (3.7%), representing different E. hormaechei subspecies. Most of the isolates contained the Tn125A variant of the K. pneumoniae ST11 lineage, and several had Tn125F of the ST147. Individual E. hormaechei genotypes represented various epidemiological situations, from sporadic cases to single-hospital, city and regional outbreaks, including one caused by ST89 organisms with 82 cases in 17 centres. Acquisitions of the Tn125A/Tn125F determinants by the E. hormaechei strains occurred around 10 times and were plasmid-mediated, with a significant plasmid rearrangement in case of Tn125F. CONCLUSIONS: The increase in E. hormaechei NDM-1 cases in Poland is a consequence of the uncontrolled spread of NDM-1-producing K. pneumoniae genotypes. Several E. hormaechei lineages have acquired NDM-encoding plasmids in different locales which started 'secondary' progressive outbreaks.


Subject(s)
Klebsiella Infections , Klebsiella pneumoniae , Anti-Bacterial Agents/pharmacology , Disease Outbreaks , Enterobacter , Humans , Klebsiella Infections/epidemiology , Microbial Sensitivity Tests , Multilocus Sequence Typing , Plasmids , beta-Lactamases/genetics
4.
J Antimicrob Chemother ; 76(9): 2273-2284, 2021 08 12.
Article in English | MEDLINE | ID: mdl-34179963

ABSTRACT

OBJECTIVES: To identify key factors of the expansion of metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa (MPPA) in Poland, focusing on the role of clonal epidemic(s). METHODS: MPPA isolates were typed by PFGE, followed by MLST. blaVIM/IMP MBL genes were amplified and sequenced within class 1 integrons. Their location was assessed by S1 nuclease-hybridization assays. Short-read WGS was performed, and genomes were subjected to SNP-based phylogenetic and resistome analyses. RESULTS: Of 1314 MPPA isolates collected in 2005-15 from 212 hospitals, 454 representatives were selected. The isolates belonged to 120 pulsotypes and 52 STs, of which ST235 (∼31%), ST111 (∼17%), ST273 (∼16%) and ST654 (∼9%) prevailed, followed by ST244, ST17, ST395, ST175 and ST1567. The isolates produced seven VIM variants (97.5%) and four IMPs encoded by 46 integrons, most of which were observed only or mainly in Poland. Around 60% of the isolates resulted from (inter)regional clonal outbreaks of 10 individual ST235, ST111, ST273 and ST654 genotypes. The phylogenetic analysis of 163 genomes revealed heterogeneity of ST235 and ST111 populations, arising from transnational circulation and on-site differentiation of several clades/branches. Contrarily, ST273 and ST654 formed relatively homogeneous and apparently Poland-specific lineages, and a unique ST273 genotype with integron In249 was the most expansive organism. CONCLUSIONS: Together with a previous report on self-transmissible In461-carrying IncP-2-type plasmids, this study revealed the molecular/genomic background of the rapid MPPA increase in Poland in 2001-15, evidencing multi-clonal spread as its leading factor. Numerous novel/specific MPPA characteristics were identified.


Subject(s)
Pseudomonas Infections , Pseudomonas aeruginosa , Humans , Anti-Bacterial Agents , beta-Lactamases/genetics , Genomics , Genotype , Microbial Sensitivity Tests , Molecular Epidemiology , Multilocus Sequence Typing , Phylogeny , Poland/epidemiology , Pseudomonas aeruginosa/genetics , Pseudomonas Infections/epidemiology
6.
J Antimicrob Chemother ; 74(11): 3199-3204, 2019 11 01.
Article in English | MEDLINE | ID: mdl-31406993

ABSTRACT

OBJECTIVES: In 2015 and 2016 Poland recorded rapid proliferation of New Delhi MBL (NDM)-producing Enterobacterales, with at least 470 and 1780 cases, respectively. We addressed the roles of the Klebsiella pneumoniae ST11 NDM-1 outbreak genotype, already spreading in 2012-14, and of newly imported organisms in this increase. METHODS: The study included 2136 NDM-positive isolates identified between April 2015 and December 2016, following transfer of patients with K. pneumoniae ST147 NDM-1 from Tunisia to Warsaw in March 2015. The isolates were screened by PCR mapping for variants of blaNDM-carrying Tn125-like elements. Selected isolates were typed by PFGE and MLST. NDM-encoding plasmids were analysed by nuclease S1/hybridization, transfer assays, PCR-based replicon typing and PCR mapping. RESULTS: The organisms were mainly K. pneumoniae containing the Tn125A variant of the ST11 epidemic lineage (n = 2094; ∼98%). Their representatives were of the outbreak pulsotype and ST11, and produced NDM-1, encoded by specific IncFII (pKPX-1/pB-3002cz)-like plasmids. The isolates were recovered in 145 healthcare centres in 13/16 administrative regions, predominantly the Warsaw area. The 'Tunisian' genotype K. pneumoniae ST147 NDM-1 Tn125F comprised 18 isolates (0.8%) from eight institutions. The remaining 24 isolates, mostly K. pneumoniae and Escherichia coli of diverse STs, produced NDM-1 or NDM-5 specified by various Tn125 derivatives and plasmids. CONCLUSIONS: The K. pneumoniae ST11 NDM-1 outbreak has dramatically expanded in Poland since 2012, which may bring about a countrywide endemic situation in the near future. In addition, the so-far limited K. pneumoniae ST147 NDM-1 outbreak plus multiple NDM imports from different countries were observed in 2015-16.


Subject(s)
Communicable Diseases, Imported/epidemiology , Klebsiella Infections/epidemiology , Klebsiella pneumoniae/classification , beta-Lactamases/genetics , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , Communicable Diseases, Imported/microbiology , Cross Infection/epidemiology , Disease Outbreaks , Endemic Diseases , Genome, Bacterial , Genotype , Humans , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/enzymology , Microbial Sensitivity Tests , Multilocus Sequence Typing , Poland/epidemiology , Tunisia/epidemiology
8.
Clin Microbiol Infect ; 25(11): 1399-1407, 2019 Nov.
Article in English | MEDLINE | ID: mdl-30980927

ABSTRACT

OBJECTIVES: High-quality diagnosis of bloodstream infections (BSI) is important for successful patient management. As knowledge on current practices of microbiological BSI diagnostics is limited, this project aimed to assess its current state in European microbiological laboratories. METHODS: We performed an online questionnaire-based cross-sectional survey comprising 34 questions on practices of microbiological BSI diagnostics. The ESCMID Study Group for Bloodstream Infections, Endocarditis and Sepsis (ESGBIES) was the primary platform to engage national coordinators who recruited laboratories within their countries. RESULTS: Responses were received from 209 laboratories in 25 European countries. Although 32.5% (68/209) of laboratories only used the classical processing of positive blood cultures (BC), two-thirds applied rapid technologies. Of laboratories that provided data, 42.2% (78/185) were able to start incubating BC in automated BC incubators around-the-clock, and only 13% (25/192) had established a 24-h service to start immediate processing of positive BC. Only 4.7% (9/190) of laboratories validated and transmitted the results of identification and antimicrobial susceptibility testing (AST) of BC pathogens to clinicians 24 h/day. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry from briefly incubated sub-cultures on solid media was the most commonly used approach to rapid pathogen identification from positive BC, and direct disc diffusion was the most common rapid AST method from positive BC. CONCLUSIONS: Laboratories have started to implement novel technologies for rapid identification and AST for positive BC. However, progress is severely compromised by limited operating hours such that current practice of BC diagnostics in Europe complies only partly with the requirements for optimal BSI management.


Subject(s)
Diagnostic Tests, Routine/methods , Microbiological Techniques/methods , Molecular Diagnostic Techniques/methods , Sepsis/diagnosis , Cross-Sectional Studies , Europe , Humans
10.
J Antimicrob Chemother ; 73(10): 2675-2681, 2018 10 01.
Article in English | MEDLINE | ID: mdl-29986025

ABSTRACT

Objectives: To analyse VIM/IMP-type MBL-producing Enterobacteriaceae isolates identified in Poland during 2006-12. Methods: Isolates were typed by PFGE, followed by MLST. blaVIM/IMP genes were amplified and sequenced within class 1 integrons. Their plasmidic versus chromosomal location was assessed by nuclease S1 and I-CeuI plus hybridization experiments. Plasmids were characterized by transfer assays and PCR-based replicon typing. Results: One hundred and nineteen VIM/IMP-positive Enterobacteriaceae cases were reported in Poland from the first case in 2006 until 2012. The patients were in 54 hospitals and were infected or colonized by 121 organisms, including Enterobacter cloacae complex (n = 64), Klebsiella oxytoca (n = 23), Serratia marcescens (n = 20) and Klebsiella pneumoniae (n = 11). The isolates represented numerous pulsotypes and mainly original STs, and carried eight integrons with blaVIM-1-like genes (blaVIM-1/-4/-28/-37/-40; n = 101), three with blaVIM-2 variants (blaVIM-2/-20; n = 17) and one with blaIMP-19 (n = 3). Six integrons were new, and five and two formed prevalent families of In238-like (n = 96) and In1008-like (n = 16) elements, respectively. In238 (aacA4-blaVIM-4rpt) and In1008 (blaVIM-2-aacA4) had been originally observed in Polish Pseudomonas aeruginosa, suggestive of their transfer to enterobacteria, followed by spread and diversification. Four organisms have disseminated inter-regionally, i.e. Enterobacter hormaechei ST90 with plasmidic In238/In238a integrons (n = 36), K. oxytoca ST145 with a chromosomal In237-like element (n = 18) and two subclones of E. hormaechei ST89 with In1008- or In238-type variants (n = 8 and n = 7, respectively). Conclusions: The epidemiology of VIM/IMP-producing Enterobacteriaceae in Poland has revealed a remarkable number of specific or novel characteristics of the organisms, with some possible links to other mid-southern European countries.


Subject(s)
Carbapenem-Resistant Enterobacteriaceae/isolation & purification , Enterobacteriaceae Infections/epidemiology , Enterobacteriaceae/enzymology , Klebsiella oxytoca/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Bacterial Typing Techniques , Carbapenem-Resistant Enterobacteriaceae/genetics , Carbapenems/pharmacology , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , Enterobacteriaceae/drug effects , Enterobacteriaceae/genetics , Enterobacteriaceae Infections/microbiology , Epidemics , Humans , Integrons/genetics , Klebsiella oxytoca/genetics , Microbial Sensitivity Tests , Multilocus Sequence Typing , Plasmids/genetics , Poland/epidemiology , beta-Lactamases/genetics
11.
J Antimicrob Chemother ; 73(3): 620-625, 2018 03 01.
Article in English | MEDLINE | ID: mdl-29237086

ABSTRACT

Objectives: To analyse OXA-48 (OXA-48/181)-type carbapenemase-producing Enterobacteriaceae reported in Poland from 2013 until January 2017. Methods: Bacterial isolates were typed by PFGE and MLST. Genes coding for OXA-48/181 types and other ß-lactamases were amplified and sequenced. Mobile elements with blaOXA-48/181-like genes were PCR mapped. blaOXA-48/181-carrying plasmids were characterized by nuclease S1-hybridization profiling, transfer assays and PCR-based replicon typing, while the chromosomal location of the genes was confirmed by the I-CeuI analysis. Results: Fifty-four isolates from 52 patients in 20 hospitals (14 cities) were included, in 14 cases having probable foreign origins indicated. The organisms were genetically diverse and represented numerous pandemic clones, including Klebsiella pneumoniae ST395 (n = 23), ST11, ST15 and ST101, Escherichia coli ST38, ST410 and ST648, and Enterobacter cloacae complex ST78. These produced OXA-48 (n = 49), OXA-181 (n = 4) or OXA-232 (n = 1). One of five K. pneumoniae ST395 pulsotypes caused a multicentre outbreak with 18 cases, which significantly contributed to the total number of patients. Depending on the variant, the blaOXA-48/181-like genes were parts of the Tn1999-, Tn2013- or Tn2016-like transposons, with blaOXA-48 found in an ISEcp1-associated module (Tn2016-like) for the first time. Three genotypes, including E. coli ST38, had chromosomal blaOXA-48 genes, while others carried transmissible IncL (∼60 kb; blaOXA-48; n = 30), IncM (∼80-95 kb; blaOXA-48; n = 4), IncX3 (∼50 kb; blaOXA-181; n = 4) or non-typeable (∼90-160 kb; blaOXA-48 or blaOXA-232) plasmids. Conclusions: Even though OXA-48/181 producers seem to occur infrequently in Poland, their epidemiology has been marked by various phenomena, namely multiple imports, several limited transmissions plus one larger clonal outbreak, and possible plasmid transfers.


Subject(s)
Bacterial Proteins/biosynthesis , Enterobacteriaceae Infections/epidemiology , Enterobacteriaceae/enzymology , Enterobacteriaceae/genetics , beta-Lactamases/biosynthesis , Bacterial Typing Techniques , Enterobacteriaceae/classification , Escherichia coli/enzymology , Escherichia coli Infections/epidemiology , Escherichia coli Proteins/biosynthesis , Humans , Klebsiella Infections/epidemiology , Klebsiella pneumoniae , Microbial Sensitivity Tests , Multilocus Sequence Typing , Poland/epidemiology
12.
Eur J Clin Microbiol Infect Dis ; 36(11): 2281-2287, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28744664

ABSTRACT

The aim of this study was to evaluate the Carba NP test (and CarbAcineto) for the detection of carbapenemases in Enterobacteriaceae, Pseudomonas spp. and Acinetobacter spp., and to assess its usefulness in the routine work of the National Reference Centre for Susceptibility Testing (NRCST) in Poland. The evaluation of the Carba NP/CarbAcineto tests was carried out on a group of 81 Enterobacteriaceae, Pseudomonas spp. and Acinetobacter spp. isolates producing KPC-, NDM-, VIM-, IMP- or OXA-48, -23, -24/40, -58-type carbapenemases, and on 26 carbapenemase-negative strains cultivated on a broad panel of microbiological media. Subsequently, the performance of the Carba NP/CarbAcineto tests was assessed on 1282 isolates of Enterobacteriaceae, Pseudomonas spp. and Acinetobacter spp. from Polish hospitals, submitted to the NRCST during a 9-month period in 2014. The Carba NP/CarbAcineto results were compared with other phenotypic tests and/or polymerase chain reaction (PCR). The impact of the media on the results of the Carba NP/CarbAcineto tests was observed, with the Columbia blood agar yielding the highest sensitivity and clarity of the results. Furthermore, the Carba NP/CarbAcineto tests were included in the NRCST routine procedure for carbapenemase identification. The sensitivity and specificity of the Carba NP test were 95.8% and 93.3%, respectively, for Enterobacteriaceae, and 97.5% and 99.0%, respectively, for Pseudomonas spp. The sensitivity of the CarbAcineto test for Acinetobacter spp. was 88.9%. This study confirmed the usefulness of the Carba NP/CarbAcineto tests for the rapid detection of various types of carbapenemases.


Subject(s)
Acinetobacter/enzymology , Bacterial Proteins/metabolism , Drug Resistance, Multiple, Bacterial/physiology , Enterobacteriaceae/enzymology , Pseudomonas/enzymology , beta-Lactamases/metabolism , Acinetobacter/drug effects , Acinetobacter/isolation & purification , Enterobacteriaceae/drug effects , Enterobacteriaceae/isolation & purification , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction , Pseudomonas/drug effects , Pseudomonas/isolation & purification , Sensitivity and Specificity
13.
Eur J Clin Microbiol Infect Dis ; 36(7): 1279-1286, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28197728

ABSTRACT

The significant increase of the linezolid-resistant enterococci (LRE) has been observed in Polish hospitals since 2012 and our study aimed at elucidating the possible reasons for this phenomenon. Polish LRE isolates were analysed by multilocus-sequence typing (MLST) and multiple locus variable-number tandem repeat (VNTR) analysis (MLVA), polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) to establish clonal relatedness and mechanism of linezolid resistance, respectively. Fifty analysed LRE (2008-2015) included mostly Enterococcus faecium (82%) and Enterococcus faecalis (16%). Enterococcus faecium belonged to the hospital-adapted lineages 17/18 and 78, while E. faecalis isolates represented ST6, a hospital-associated type, and ST116, found in both humans and food-production animals. The G2576T 23S rRNA mutation was the most frequent (94%) mechanism of linezolid/tedizolid resistance of LRE. None of the isolates carried the plasmid-associated gene of Cfr methyltransferase, whereas optrA, encoding the ABC-type drug transporter, was identified in two E. faecalis isolates. In these isolates, optrA was located on a plasmid, transferable to both E. faecium and E. faecalis, whose partial (36.3 kb) sequence was 100% identical to the pE394 plasmid, identified previously in China in both clinical and farm animal isolates. The optrA-E. faecium transconjugant displayed a significant growth deficiency, in contrast to the optrA-E. faecalis. Our study indicates the role of mutation acquisition by hospital-adapted clones of enterococci as a major driver of increasing resistance to linezolid and tedizolid. Transferability and apparent lack of a biological cost of resistance suggest that E. faecalis may be a natural reservoir of optrA, an emerging mechanism of oxazolidinone resistance.


Subject(s)
Anti-Bacterial Agents/pharmacokinetics , Drug Resistance, Bacterial , Enterococcus faecalis/drug effects , Enterococcus faecium/drug effects , Gram-Positive Bacterial Infections/epidemiology , Gram-Positive Bacterial Infections/microbiology , Linezolid/pharmacology , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Enterococcus faecalis/classification , Enterococcus faecalis/genetics , Enterococcus faecalis/isolation & purification , Enterococcus faecium/classification , Enterococcus faecium/genetics , Enterococcus faecium/isolation & purification , Gene Transfer, Horizontal , Hospitals , Humans , Membrane Transport Proteins/genetics , Minisatellite Repeats , Multilocus Sequence Typing , Mutation , Plasmids/analysis , Poland/epidemiology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , RNA, Ribosomal, 23S/genetics
14.
Eur J Clin Microbiol Infect Dis ; 36(2): 313-328, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27752789

ABSTRACT

The objective of this study was to investigate the antimicrobial resistance, Tn1546 transposon variability and plasmid diversity among Polish vancomycin-resistant Enterococcus faecium (VREfm) isolates of VanA phenotype in the context of their clonal structure. Two hundred sixteen clinical VREfm isolates collected between 1997 and 2010 were studied by antimicrobial susceptibility testing, MLST, MLVA and detection of IS16, esp Efm, pilA, intA and plasmid-specific genes by PCR. Tn1546 structure was revealed by overlapping PCR and sequencing. Selected isolates were subjected to PFGE-S1 and Southern hybridization analyses. The vast majority of the isolates (95.8 %) belonged to lineages 17/18 (during the whole study period 1997-2010) and 78 (mostly in 2006-2010) of hospital-adapted meroclone of E. faecium. All isolates displayed a multi-drug resistance phenotype. Twenty-eight Tn1546 types (including 26 novel ones) were associated with eight different ISs (IS1216, IS1251, ISEfa4, ISEfa5, ISEfm2, ISEf1, IS3-like, ISEfm1-like). The vanA-determinant was typically located on plasmids, which most commonly carried rep2pRE25, rep17pRUM, rep18pEF418, rep1pIP501, ω-ε-ζ and axe-txe genes. VanA isolates from 1997-2005 to 2006-2010 differed in clonal composition, prevalence of gentamicin- and tetracycline-resistance and plasmidome. Our analysis revealed high complexity of Tn1546-type transposons and vanA-plasmids, and suggested that diverse genetic events, such as conjugation transfer, recombination, chromosomal integration and DNA mutations shaped the structure of these elements among Polish VREfm.


Subject(s)
Bacterial Proteins/genetics , Carbon-Oxygen Ligases/genetics , DNA Transposable Elements , Enterococcus faecium/genetics , Genetic Variation , Plasmids , Enterococcus faecium/isolation & purification , Evolution, Molecular , Genotyping Techniques , Gram-Positive Bacterial Infections/microbiology , Humans , Microbial Sensitivity Tests , Molecular Typing , Nucleic Acid Hybridization , Poland , Polymerase Chain Reaction , Sequence Analysis, DNA
16.
Eur J Clin Microbiol Infect Dis ; 35(5): 847-56, 2016 May.
Article in English | MEDLINE | ID: mdl-26946510

ABSTRACT

The aim of this study was to investigate human invasive isolates of enterococci, obtained through prospective surveillance in Poland. The consecutive enterococcal isolates were collected in 30 hospitals between May 2010 and June 2011, and studied by species identification, antimicrobial susceptibility testing and, for Enterococcus faecium by detection of markers specific for the hospital meroclone, multilocus VNTR analysis (MLVA) and multilocus sequence typing (MLST). Additionally, the genomic difference regions (GDRs) characteristic for lineage 78 were searched by PCR. Among 259 isolates, a nearly equal number of Enterococcus faecalis (n = 140; 54.1 %) and E. faecium (n = 112; 43.2 %) was found. The observed 14-day mortality rate of infected patients reached 18.1 %. All isolates were susceptible to linezolid and daptomycin. High-level aminoglycoside resistance occurred in over 50 % of isolates. Vancomycin resistance mediated by vanA or vanB was detected in 7.1 % of E. faecium; 71.4 % of isolates were multidrug resistant. E. faecium isolates ubiquitously carried molecular markers of hospital-associated meroclone (IS16, esp(Efm), intA of ICEEfm1) and multilocus sequence typing showed the domination of representatives of lineages 78 and 17/18 (52.7 % and 46.4 %, respectively). Isolates of lineage 78 were significantly enriched in all the GDRs studied. The recent spread of E. faecium from this lineage contributed to the observed increase of E. faecium in enterococcal invasive infections in hospitals in Poland.


Subject(s)
Enterococcus/classification , Gram-Positive Bacterial Infections/epidemiology , Gram-Positive Bacterial Infections/microbiology , Adolescent , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/pharmacology , Biofilms , Child , Child, Preschool , Cross Infection , Enterococcus/drug effects , Enterococcus/genetics , Female , Humans , Infant , Infant, Newborn , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Typing , Multilocus Sequence Typing , Patient Outcome Assessment , Poland/epidemiology , Population Surveillance , Young Adult
17.
J Antimicrob Chemother ; 71(1): 85-91, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26386745

ABSTRACT

OBJECTIVES: The objective of this study was to characterize New Delhi metallo-ß-lactamase (NDM)-producing Enterobacteriaceae isolates reported in Poland in 2012-14. METHODS: Representative isolates were typed by PFGE and MLST. NDM and other ß-lactamase genes were amplified and sequenced. Plasmids with blaNDM genes were analysed by nuclease S1 plus hybridization profiling, by transfer assays and by PCR-based replicon typing. The blaNDM genetic context was studied by PCR mapping assays. RESULTS: Of 374 cases of infection/colonization with NDM-positive Enterobacteriaceae identified in 2012-14, 370 cases in 40 hospitals, 10 outpatient clinics and 1 nursing home were associated with a Klebsiella pneumoniae outbreak with epicentres in Poznan and Warsaw. The outbreak strain of K. pneumoniae ST11 was similar to an isolate from the Czech Republic from 2013. Like the Czech strain, many of the isolates had two blaNDM-1-carrying IncFII- and IncR-type plasmids of variable size, sharing a blaNDM-1-containing segment. The early isolates also produced CTX-M-15 co-encoded by the IncR-type plasmids, and differentiated later by extensive plasmid rearrangements. Four other NDM cases were reported in 2013, three being associated with arrivals from Montenegro, India or Afghanistan. The Indian Escherichia coli ST448 NDM-5 isolate revealed similarity to a recent isolate from Spain, including the blaNDM genetic context observed previously in E. coli strains in Poland and France (of Congolese and Indian origins, respectively). The Afghani Proteus mirabilis was the second isolate of this species with a chromosomal blaNDM-1 location. CONCLUSIONS: The largest NDM outbreak in a non-endemic country has been observed, being an alarming phenomenon in resistance epidemiology in Poland.


Subject(s)
Disease Outbreaks , Klebsiella Infections/epidemiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/isolation & purification , beta-Lactamases/genetics , Adolescent , Adult , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Female , Gene Transfer, Horizontal , Genotype , Humans , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/genetics , Male , Middle Aged , Molecular Typing , Nucleic Acid Hybridization , Plasmids/analysis , Poland/epidemiology , Polymerase Chain Reaction , Sequence Analysis, DNA
18.
Euro Surveill ; 20(23)2015 Jun 11.
Article in English | MEDLINE | ID: mdl-26084313

ABSTRACT

We describe the introduction of NDM-1-producing Klebsiella pneumoniae ST147 and Escherichia coli ST410, and OXA-48-producing K. pneumoniae ST101 strains to Poland by two patients transported to the country after hospitalisation in Tunisia. The patients had gunshot wounds following the terrorist attack in the Bardo National Museum in Tunis in March 2015. Our report reinforces the need for microbiological screening of patients returning from travel on admission to healthcare institutions, especially following hospitalisation in countries where carbapenemase-producing Enterobacteriaceae are endemic.


Subject(s)
Enterobacteriaceae Infections/microbiology , Escherichia coli/enzymology , Escherichia coli/isolation & purification , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/isolation & purification , Oxytocin/analogs & derivatives , beta-Lactamases/metabolism , Adult , Anti-Bacterial Agents/pharmacology , Enterobacteriaceae/classification , Enterobacteriaceae/enzymology , Enterobacteriaceae/genetics , Enterobacteriaceae/isolation & purification , Escherichia coli Infections/diagnosis , Escherichia coli Infections/drug therapy , Humans , Klebsiella Infections/diagnosis , Klebsiella Infections/drug therapy , Male , Middle Aged , Multilocus Sequence Typing , Oxytocin/genetics , Oxytocin/metabolism , Poland , Polymerase Chain Reaction , Travel , Treatment Outcome , Tunisia , beta-Lactamases/genetics
19.
Antimicrob Agents Chemother ; 58(2): 1203-7, 2014.
Article in English | MEDLINE | ID: mdl-24247128

ABSTRACT

Poland's first Enterobacteriaceae isolate producing the New Delhi metallo-ß-lactamase (NDM) was identified in August 2011. Escherichia coli sequence type ST410 NDM-1 was cultured from a critically ill patient who had been transferred directly from the Congo. The blaNDM-1 gene was carried by conjugative IncFII-type plasmid pMC-NDM (87,619 bp), which showed structural similarity to plasmid pGUE-NDM, which was identified earlier in France in an E. coli ST131 isolate of Indian origin.


Subject(s)
Escherichia coli Infections/microbiology , Escherichia coli/genetics , Plasmids/chemistry , beta-Lactamases/genetics , Anti-Bacterial Agents/therapeutic use , Drug Resistance, Multiple, Bacterial , Escherichia coli/isolation & purification , Escherichia coli Infections/drug therapy , Escherichia coli Infections/pathology , Fatal Outcome , Gene Expression , Humans , Male , Middle Aged , Multilocus Sequence Typing , Poland
20.
Med Dosw Mikrobiol ; 52(4): 317-26, 2000.
Article in Polish | MEDLINE | ID: mdl-11286173

ABSTRACT

The ability of lysogenization was examined of 50 S. intermedius strains and of 77 strains belonging to 14 different species of coagulase-negative staphylococci using 8 enterotoxin A converting bacteriophages isolated from S. aureus. All the examined bacteriophages showed lytic activity against at least 1 of 11 susceptible strains of S. intermedius to them. Lytic activity towards coagulase-negative staphylococci was observed for 6 of 8 examined bacteriophages. Two bacteriophages were active against 1 of 9 examined S. capitis strains, one against 1 of 11 examined S. haemolyticus strains, four against 1 of 6 examined S. lugdunensis strains, three against 1 of 6 examined S. warneri strains and one against 1 of 5 examined S. xylosus strains. Lysogenization with bacteriophage f421-1 able to convert positively enterotoxin A and staphylokinase and negatively beta-haemolysin of one S. intermedius strain was successful. S. intermedius lysogenized with phi 421-1 was able to produce both enterotoxin A and staphylokinase and lost ability to produce beta-haemolysin. Our results showed a broad lytic spectrum and interspecies host range of some S. aureus bacteriophages and the ability of interspecies transfer of bacteriophages between S. aureus and S. intermedius.


Subject(s)
Lysogeny , Staphylococcus Phages/physiology , Staphylococcus/metabolism , Staphylococcus/virology , Enterotoxins/biosynthesis , Hemolysin Proteins/biosynthesis , Metalloendopeptidases/biosynthesis , Species Specificity , Staphylococcus aureus/metabolism , Staphylococcus aureus/virology
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