Klebsiella pneumoniae infections have always been an important problem in public health, but today, the increasing resistance of these bacteria to antibiotics due to β-lactamases production has renewed interest in K. pneumoniae infections. The aim of the study was to present a case of a neurosurgical patient with multidrug-resistant K. pneumoniae ST11 infection after craniectomy. Four K. pneumoniae isolates from various clinical materials of the patient undergone identification and susceptibility testing with the Vitek2 system. Tests for β-lactamases production were performed according to EUCAST guidelines. Strains were analyzed for bla genes responsible for β-lactamase production (blaTEM, blaSHV, blaCTX-M, blaVIM, blaIMP, blaNDM, blaKPC, blaOXA-48) using PCR. Moreover, the genetic relatedness of these isolates was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). All tested strain presented multidrug resistance. The highest susceptibility was observed for imipenem, meropenem, and ertapenem. The strain isolated from the nervous system was ESBL-positive with blaSHV-11, blaTEM-1, and blaCTX-M-15 genes. Additionally, the strain from urine was blaKPC-3-positive. Molecular typing revealed that all strains belonged to the same clone and identified two PFGE profiles. The analysis of MLST allelic profile showed that tested K. pneumoniae strains belonged to ST11. Identification of ST11 K. pneumoniae as etiological factor of infection unfavorably impacts on prognosis among neurosurgical patient after craniectomy.

• MeSH
• antibakteriální látky farmakologie   terapeutické užití   MeSH
• beta-laktamasy genetika   MeSH
• fatální výsledek MeSH
• infekce bakteriemi rodu Klebsiella diagnostické zobrazování   farmakoterapie   mikrobiologie   MeSH
• Klebsiella pneumoniae klasifikace   účinky léků   enzymologie   MeSH
• kraniotomie škodlivé účinky   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• mladý dospělý MeSH
• mnohočetná bakteriální léková rezistence * MeSH
• multilokusová sekvenční typizace MeSH
• počítačová rentgenová tomografie MeSH
• pulzní gelová elektroforéza MeSH
• techniky typizace bakterií MeSH
• Check Tag
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Oxacillinases (OXA) have been mostly described in Enterobacteriaceae, Acinetobacter, and Pseudomonas species. Recent years have witnessed an increased prevalence of intrinsic and/or acquired β-lactamase-producing Acinetobacter in food-producing animals. This study was conducted to assess the prevalence of OXA among selected bacterial species and to characterize these enzymes by in silico analysis. Screening of OXA was performed by PCR amplification using specific pairs of oligonucleotides. Overall, 40 pairs of primers were designed, of which 6 were experimentally tested in vitro. Among 49 bacterial isolates examined, the presence of blaOXA-1-like genes was confirmed in 20 cases (41%; 19 times in Klebsiella pneumoniae and once in Enterobacter cloacae). No OXA were found in animal isolates. The study results confirmed the specificity of the designed oligonucleotide pairs. Furthermore, the designed primers were found to possess the ability to specifically detect 90.2% of all OXA. These facts suggest that the in silico and in vitro tested primers could be used for single or multiplex PCR to screen for the presence of OXA in various bacteria, as well as to monitor their spread. At the same time, the presence of conserved characteristic amino acids and motifs was confirmed by in silico analysis of sequences of representative members of OXA.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• DNA primery chemická syntéza   metabolismus   MeSH
• Enterobacter cloacae klasifikace   účinky léků   enzymologie   genetika   MeSH
• Escherichia coli klasifikace   účinky léků   enzymologie   genetika   MeSH
• exprese genu MeSH
• fylogeneze MeSH
• gramnegativní bakteriální infekce diagnóza   epidemiologie   mikrobiologie   veterinární   MeSH
• Klebsiella pneumoniae klasifikace   účinky léků   enzymologie   genetika   MeSH
• kur domácí mikrobiologie   MeSH
• lidé MeSH
• maso mikrobiologie   MeSH
• mikrobiální testy citlivosti MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• peniciliny farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Dissemination of carbapenemase-producing Klebsiella pneumoniae along with 16S rRNA methyltransferase (16S-RMTase) has been caused as a great concern for healthcare settings. The aim of this study was to investigate the prevalence of resistance genes among K. pneumoniae isolates. During October 2015 to February 2016, 30 non-duplicative K. pneumoniae strains were isolated from clinical specimens in a burn center in Kerman, Iran. Antibiotic susceptibility tests of isolates, carbapenemase, extended-spectrum beta-lactamases (ESBLs) and AmpC beta-lactamase-producing isolates were determined by phenotypic methods. The beta-lactamase, oqxA/B efflux pumps, qnr A, B, S, 16S-RMTase (rmt A, B, and C), and mcr-1 resistance genes were determined by PCR. Enterobacterial repetitive intergenic consensus (ERIC)-PCR was used for molecular typing. According to our findings, tigecycline has been shown the most active agent against K. pneumoniae isolates. Antibiotic resistance genes, blaTEM-1, blaSHV-12, blaCTX-M-15, blaCTX-M-2, blaNDM-1, blaFOX, blaMOX, blaEBC, blaACC, blaCIT, rmtC, qnrB, qnrS, oqxA, and oqxB, were detected in 11 (36.7%), 13 (43.3%), 11 (36.6%), 5 (16.6%), 9 (30%), 1 (3.3%), 1 (3.3%), 1 (3.3%), 1 (3.3%), 2 (6.7%), 1 (3.3%), 9 (30%), 2 (6.7%), 18 (60%), and 13 (43.3%) of isolates, respectively. The blaNDM-1 with rmtC was simultaneously observed in one isolate. ERIC-PCR results revealed 25 distinct patterns in eight clusters (A-H) and five singletons. This study highlights the high prevalence of blaNDM and emergence of rmtC among carbapenem-resistant K. pneumoniae. The resistance genes are often co-located on the conjugative plasmids, so it might be the reason of the rapid spread of them. The prevalence of multidrug-resistant K. pneumoniae isolates limits the available treatment options and presents tremendous challenges to public health.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence genetika   MeSH
• bakteriální proteiny genetika   MeSH
• beta-laktamasy genetika   MeSH
• Enterobacteriaceae rezistentní na karbapenemy účinky léků   genetika   izolace a purifikace   MeSH
• infekce bakteriemi rodu Klebsiella mikrobiologie   MeSH
• Klebsiella pneumoniae klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• lidé MeSH
• methyltransferasy genetika   MeSH
• mikrobiální testy citlivosti MeSH
• molekulární typizace MeSH
• plazmidy genetika   MeSH
• popáleninové jednotky * MeSH
• rozptýlené repetitivní sekvence genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Írán MeSH

Intensive fish farming systems have led to increase in disease incidence, due to higher stocking density, high organic matter levels, and poor quality of the aquatic environment. Diseased fish samples showing hemorrhages and reddish lesions were collected from different freshwater fish farms located at three different districts of West Bengal, India (Burdwan, North 24 Parganas, and Nadia). The present study was conducted to evaluate the genetic diversity of ten different Klebsiella pneumoniae strains isolated from different infected freshwater fish samples based on 16S rRNA gene sequence analysis. Primarily, Klebsiella-specific media was used for the isolation and characterization of Klebsiella pneumoniae. Further, through a biochemical test, all the strains were confirmed as K. pneumoniae. PCR analysis of 16S-23S internal transcribed spacer (PCR ribotyping) was carried out to study the species variation within different Klebsiella pneumoniae isolates. For all the isolates, a conserved PCR ribotype pattern was observed while differing from other bacterial species. Phylogenetic study showed the high degree of homology with diverse source of other strains. The multiple antibiotic resistance (MAR) values of the present study for the isolates were found to be 0.468. MAR value above 0.2 indicates that the source of isolation was highly contaminated with antibiotics. Based on the 16S rRNA gene sequence analysis, the present study revealed the genetic diversity of Klebsiella pneumoniae isolated from the different diseased fish farms of West Bengal. All the strains were found to be hypermucoviscous and multidrug-resistant, thus making it pathogenic towards the host organisms. Further, the study revealed a high prevalence of K. pneumoniae in aquaculture farms, representing a risk towards successful aquaculture.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální geny genetika   MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• hemolýza MeSH
• infekce bakteriemi rodu Klebsiella epidemiologie   mikrobiologie   veterinární   MeSH
• kapři mikrobiologie   MeSH
• Klebsiella pneumoniae klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• mezerníky ribozomální DNA genetika   MeSH
• mikrobiální testy citlivosti MeSH
• mnohočetná bakteriální léková rezistence * MeSH
• nemoci ryb mikrobiologie   MeSH
• prevalence MeSH
• ribotypizace MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• vodní hospodářství MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Indie MeSH

Sequence type 11 Klebsiella pneumoniae, coproducing NDM-1 and VIM-1 metallo-β-lactamases, were isolated in a Greek hospital. blaNDM-1 was part of a Tn125 derivative, located on an ~90-kb plasmid similar to the NDM-1-encoding plasmid pB-3002cz. blaVIM-1 was located in an In-e541-like integron, carried on a multireplicon (IncA/C and IncR) plasmid of ~180kb.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• infekce bakteriemi rodu Klebsiella farmakoterapie   mikrobiologie   MeSH
• karbapenemy farmakologie   MeSH
• Klebsiella pneumoniae klasifikace   genetika   izolace a purifikace   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• nemocnice MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Řecko MeSH

Národní referenční laboratoř pro antibiotika, Státní zdravotní ústav, Praha, zaznamenala výskyt nově popsaného druhu Klebsiella variicola v rámci surveillance antibiotické rezistence invazivních kmenů Klebsiella pneumoniae. Blízká příbuznost a případná záměna s K. pneumoniae může být příčinou podcenění významu tohoto druhu u humánních klinických onemocnění.

The National Reference Laboratory for Antibiotics, National Institute of Public Health, Prague, has reported the occurrence of the newly described species, Klebsiella variicola, in the context of surveillance of antimicrobial resistance of invasive strains of Klebsiella pneumoniae. The close relationship to and possible misidentification with K. pneumoniae may cause an underestimation of the importance of this species in human clinical disease.

• Klíčová slova
• Klebsiella variicola,
• MeSH
• antibiotická rezistence MeSH
• epidemiologické monitorování MeSH
• infekce bakteriemi rodu Klebsiella * farmakoterapie   MeSH
• Klebsiella pneumoniae klasifikace   patogenita   účinky léků   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH

Minim typing is derived from the multi-locus sequence typing (MLST). It targets the same genes, but sequencing is replaced by high resolution melt analysis. Typing can be performed by analysing six loci (6MelT), four loci (4MelT) or using data from four loci plus sequencing the tonB gene (HybridMelT). The aim of this study was to evaluate Minim typing to discriminate extended-spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-KLPN) isolates at our hospital. In total, 380 isolates were analyzed. The obtained alleles were assigned according to both the 6MelT and 4MelT typing scheme. In 97 isolates, the tonB gene was sequenced to enable HybridMelT typing. We found that the presented method is suitable to quickly monitor isolates of ESBL-KLPN; results are obtained in less than 2 hours and at a lower cost than MLST. We identified a local ESBL-KLPN outbreak and a comparison of colonizing and invasive isolates revealed a long term colonization of patients with the same strain.

• MeSH
• beta-laktamasy sekrece   MeSH
• časové faktory MeSH
• centra terciární péče MeSH
• infekce bakteriemi rodu Klebsiella mikrobiologie   MeSH
• Klebsiella pneumoniae klasifikace   enzymologie   izolace a purifikace   MeSH
• lidé MeSH
• molekulární typizace metody   MeSH
• polymerázová řetězová reakce MeSH
• přenašečství mikrobiologie   MeSH
• sekvenční analýza DNA MeSH
• tranzitní teplota MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Carbapenemase-producing Enterobacteriaceae and Pseudomonas spp. are increasingly reported in many countries all over the world. Due to the resistance of those bacteria to almost all antibiotics (e.g.beta-lactams, aminoglycosides, fluoroquinolones),treatment options are seriously limited. In the Czech Republic, the incidence of carbapenemase-producing Enterobacteriaceae seems to be low, restricted to only three cases detected between 2009 and 2010.Here, we describe molecular typing of 15 carbapenemase-producing Klebsiella pneumoniae isolates identified in the Czech Republic during 2011. Five VIM-1-producing isolates belonging to sequence type (ST)11 and one VIM-4-producing isolate of ST1029 have been detected. blaVIM-1 and blaVIM-4 as a part of class 1 integrons were chromosomally located or carried by a plasmid belonging to A/C replicon type (blaVIM-4). KPC-3-producing isolates of ST512, recovered from six patients, caused an outbreak. Three more isolates producing KPC-2 enzyme belonged to ST258. Both blaKPCgenes were part of the Tn4401a transposon carried on plasmids of the pKpQIL type. The isolates were resistant to all antibiotics tested except colistin and/or gentamicin.Four of these 15 strains were recovered from patients repatriated to the Czech Republic from Greece and Italy. This is the first report of outbreaks caused by carbapenemase-producing Enterobacteriaceae in the Czech Republic.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny metabolismus   MeSH
• beta-laktamasy metabolismus   MeSH
• beta-laktamová rezistence MeSH
• cestování MeSH
• incidence MeSH
• infekce bakteriemi rodu Klebsiella diagnóza   enzymologie   epidemiologie   MeSH
• integrony genetika   MeSH
• karbapenemy farmakologie   MeSH
• Klebsiella pneumoniae klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• mnohočetná bakteriální léková rezistence MeSH
• molekulární typizace MeSH
• plazmidy genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Itálie MeSH
• Řecko MeSH

Resistance to carbapenems in enterobacteria is mediated by the production of several types of carbapenemases or by the decreased permeability of the outer membrane, combined with the expression of extended-spectrum beta-lactamases (ESBLs) or AmpC-like cephalosporinases. The objective of this study was to characterize carbapenem-nonsusceptible (C-NS) isolates of Klebsiella pneumoniae in the University Hospital in Plzen (Czech Republic) and compare them with carbapenem-susceptible (C-S) K. pneumoniae isolates from the same patients. Six C-NS K pneumoniae isolates from different patients were collected between January 2007 and June 2008, and from three of these patients, C-S isolates were available for the study as well. The isolates were typed by pulsed-field gel electrophoresis and multilocus sequence typing. beta-Lactamases were analyzed by isoelectric focusing, bioassay, and PCR and sequencing of bla genes. Major porin channels, OmpK35 and OmpK36, were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot; porin genes were amplified and sequenced, and their expression was assessed by reverse transcriptase-PCR. The C-NS isolates belonged to three pulsotypes and to the clone ST11, produced the SHV-5 ESBL and/or DHA-1 AmpC-type cephalosporinase, did not express OmpK36, and had a reduced expression of OmpK35. The C-S isolates differed from their C-NS counterparts only by porin expression profiles.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• infekce bakteriemi rodu Klebsiella mikrobiologie   MeSH
• karbapenemy farmakologie   MeSH
• Klebsiella pneumoniae klasifikace   účinky léků   enzymologie   genetika   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• nemocnice MeSH
• techniky typizace bakterií MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

The aim of the study was to determine the prevalence of Pseudomonas aeruginosa and Klebsiella pneumoniae strains in patients with acute leukemias, to assess their clinical significance, and to define the sources and ways of their spread using genetic analysis. Thirty-four patients were investigated during the observed period. Twenty-one strains of Pseudomonas aeruginosa and 35 strains of Klebsiella pneumoniae were isolated from patient samples. In the case of Pseudomonas aeruginosa, 47.6% of strains were identified as pathogens and caused infection. By contrast, only 4 isolates (11.4%) of Klebsiella pneumoniae could be regarded as etiological agents of bacterial infection. Based on the obtained results, Klebsiella pneumoniae strains are assumed to be of mostly endogenous origin. In the case of Pseudomonas aeruginosa strains, the proportion of identical strains detected in various patients was higher and exogenous sources were more significant. In addition, our results confirmed the ability of Pseudomonas aeruginosa strains to survive on a particular site in the hospital for a longer time.

• MeSH
• antibakteriální látky farmakologie   MeSH
• financování organizované MeSH
• hematologické nádory komplikace   MeSH
• infekce bakteriemi rodu Klebsiella epidemiologie   mikrobiologie   přenos   MeSH
• Klebsiella pneumoniae genetika   izolace a purifikace   klasifikace   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• prevalence MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   přenos   MeSH
• Pseudomonas aeruginosa genetika   izolace a purifikace   klasifikace   MeSH
• Check Tag
• lidé MeSH