Multiresistant bacteria, especially of carbapenemase-producing Enterobacteriaceae and multiresistant Pseudomonas spp. is a threat to current medicine, significantly reducing treatment options in critically ill patients. The project is focused on molecular-epidemiological typing of clinically/epidemiologically important resistance genes encoded on mobile genetic elements in Enterobacteriaceae and Pseudomonas spp., spreading in bacterial populations in health-care settings, diffusing into the environment by sewage water and colonizing/infecting wildlife, especially water birds. In the project, only bacterial isolates (not primary clinical samples) will be used. All these isolates will be collected anonymously, therefore, patients informed consent will not be a prerequisite for further molecular analysis. Obtained data will be published in international journals, popularized for microbiologists and infectious-disease specialists; and used for education of M.D., D.V.M. and Ph.D. students.

Šíření multirezistentních bakterií, především Enterobacteriaceae produkujících karbapenemázy a multirezistentních pseudomonád, představuje významné ohrožení dalšího rozvoje medicíny zejména u kriticky nemocných pacientů. Projekt je zaměřen na molekulárně-epidemiologickou analýzu klinicky/epidemiologicky významných genů rezistence a mobilních elementů, na nichž jsou kódovány, u enterobakterií a pseudomonád, jejich rozšíření v nemocničních zařízeních, difúzi do prostředí prostřednictvím odpadní vody a kolonizaci/infekci volně žijících zvířat, především vodního ptactva. Během řešení projektu budou zpracovávány pouze bakteriální izoláty (nikoliv primární pacientské vzorky). Tyto izoláty jsou anonymizovány. Z toho důvodu není vyžadován informovaný souhlas. Získaná data budou publikována v mezinárodních časopisech a popularizována pro využití mikrobiology a infekčními lékaři. Zároveň budou využita k výuce v magisterském (MUDr., MDDr., MVDr.) a doktorském (Ph.D.) studijním programu.

• MeSH
• bakteriální léková rezistence MeSH
• Enterobacteriaceae rezistentní na karbapenemy MeSH
• Enterobacteriaceae MeSH
• farmakoepidemiologie MeSH
• fluorochinolony MeSH
• karbapenemy MeSH
• Pseudomonas MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• molekulární biologie, molekulární medicína
• farmacie a farmakologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

This study evaluated the carriage of AmpC and extended-spectrum beta-lactamase (ESBL) genes and associated plasmids in faecal bacteria of Canadian corvids. Faecal samples from 449 birds in five roosting sites across Canada were analyzed using selective media, screening for AmpC and ESBL genes by PCR, and sequencing. Genomic relatedness was determined by PFGE and MLST. Plasmid mobility was studied by conjugation and transformation experiments, followed by plasmid typing. In total, 96 (21%, n = 449) cefotaxime-resistant Escherichia coli and three (0.7%) Klebsiella pneumoniae isolates were identified. ESBL genes blaCTX-M-1 (n = 3), blaCTX-M-14 (n = 2), blaCTX-M-32 (n = 2) and blaCTX-M-124 (n = 1) were detected in eight E. coli isolates, whereas blaSHV-2 (2) was found in two K. pneumoniae. E. coli isolates contained blaCMY-2 (n = 83) and blaCMY-42 (n = 1). The high genetic diversity of the isolates and presence of clinically important E. coli ST69 (n = 1), ST117 (n = 7) and ST131 (n = 1) was revealed. AmpC genes were predominantly carried by plasmids of incompatibility groups I1 (45 plasmids), A/C (10) and K (7). The plasmid IncI1/ST12 was most common and found in diverse E. coli STs in all sites. Highly diverse E. coli isolates containing AmpC and ESBL genes, including clinically important clones and emerging plasmids, are in circulation throughout Canadian wildlife.

• MeSH
• bakteriální proteiny genetika   MeSH
• beta-laktamasy genetika   MeSH
• Escherichia coli klasifikace   enzymologie   genetika   izolace a purifikace   MeSH
• feces mikrobiologie   MeSH
• Klebsiella pneumoniae enzymologie   genetika   izolace a purifikace   MeSH
• multilokusová sekvenční typizace MeSH
• plazmidy genetika   MeSH
• vrány mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Kanada MeSH

ST252 Enterobacter cloacae, producing GES-5 carbapenemase, was isolated in a Czech hospital. blaGES-5was part of a novel class 1 integron, In1406, which also included a new allele of the aadA15 gene cassette. In1406 was located on a ColE2-like plasmid, pEcl-35771cz (6953bp).

• MeSH
• bakteriální proteiny genetika   MeSH
• beta-laktamasy genetika   MeSH
• Enterobacter cloacae enzymologie   genetika   MeSH
• enterobakteriální infekce mikrobiologie   MeSH
• integrony genetika   MeSH
• lidé MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Multidrug resistant (MDR) Gram-negative bacteria have been increasingly reported in humans, companion animals and farm animals. The growing trend of plasmid-mediated resistance to antimicrobial classes of critical importance is attributed to the emergence of epidemic plasmids, rapidly disseminating resistance genes among the members of Enterobacteriaceae family. The use of antibiotics to treat humans and animals has had a significant impact on the environment and on wild animals living and feeding in human-influenced habitats. Wildlife can acquire MDR bacteria selected in hospitals, community or livestock from diverse sources, including wastewater, sewage systems, landfills, farm facilities or agriculture fields. Therefore, wild animals are considered indicators of environmental pollution by antibiotic resistant bacteria, but they can also act as reservoirs and vectors spreading antibiotic resistance across the globe. The level of resistance and reported plasmid-mediated resistance mechanisms observed in bacteria of wildlife origin seem to correlate well with the situation described in humans and domestic animals. Additionaly, the identification of epidemic plasmids in samples from different human, animal and wildlife sources underlines the role of horizontal gene transfer in the dissemination of resistance genes. The present review focuses on reports of plasmid-mediated resistance to critically important antimicrobial classes such as broad-spectrum beta-lactams and colistin in Enterobacteriaceae isolates from samples of wildlife origin. The role of plasmids in the dissemination of ESBL-, AmpC- and carbapenemase-encoding genes as well as plasmid-mediated colistin resistance determinants in wildlife are discussed, and their similarities to plasmids previously identified in samples of human clinical or livestock origin are highlighted. Furthermore, we present features of completely sequenced plasmids reported from wildlife Enterobacteriaceae isolates, with special focus on genes that could be associated with the plasticity and stable maintenance of these molecules in antibiotic-free environments.

• MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriální proteiny genetika   MeSH
• beta-laktamasy genetika   MeSH
• beta-laktamy terapeutické užití   MeSH
• gramnegativní bakterie účinky léků   genetika   patogenita   MeSH
• lidé MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• plazmidy genetika   MeSH
• přenos genů horizontální genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

• MeSH
• bakteriální proteiny genetika   MeSH
• beta-laktamasy genetika   MeSH
• Enterobacter aerogenes genetika   izolace a purifikace   MeSH
• enterobakteriální infekce mikrobiologie   veterinární   MeSH
• nemoci ptáků mikrobiologie   MeSH
• plazmidy analýza   MeSH
• vrány * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Kanada MeSH

The aim of the present study was to characterize sporadic cases and an outbreak of NDM-like-producing Enterobacteriaceae recovered from hospital settings, in Czechia. During 2016, 18 Entrobacteriaceae isolates including 10 Enterobacter cloacae complex (9 E. xiangfangensis and 1 E. asburiae), 4 Escherichia coli, 1 Kluyvera intermedia, 1 Klebsiella pneumoniae, 1 Klebsiella oxytoca, and 1 Raoultella ornithinolytica that produced NDM-like carbapenemases were isolated from 15 patients. Three of the patients were colonized or infected by two different NDM-like producers. Moreover, an NDM-4-producing isolate of E. cloacae complex, isolated in 2012, was studied for comparative purposes. All isolates of E. cloacae complex, except the E. asburiae, recovered from the same hospital, were assigned to ST182. Additionally, two E. coli belonged to ST167, while the remaining isolates were not clonally related. Thirteen isolates carried blaNDM-4, while six isolates carried blaNDM-1 (n = 3) or blaNDM-5 (n = 3). Almost all isolates carried blaNDM-like-carrying plasmids being positive for the IncX3 allele, except ST58 E. coli and ST14 K. pneumoniae isolates producing NDM-1. Analysis of plasmid sequences revealed that all IncX3 blaNDM-like-carrying plasmids exhibited a high similarity to each other and to previously described plasmids, like pNDM-QD28, reported from worldwide. However, NDM-4-encoding plasmids differed from other IncX3 plasmids by the insertion of a Tn3-like transposon. On the other hand, the ST58 E. coli and ST14 K. pneumoniae isolates carried two novel NDM-1-encoding plasmids, pKpn-35963cz, and pEsco-36073cz. Plasmid pKpn-35963cz that was an IncFIB(K) molecule contained an acquired sequence, encoding NDM-1 metallo-β-lactamase (MβL), which exhibited high similarity to the mosaic region of pS-3002cz from an ST11 K. pneumoniae from Czechia. Finally, pEsco-36073cz was a multireplicon A/C2+R NDM-1-encoding plasmid. Similar to other type 1 A/C2 plasmids, the blaNDM-1 gene was located within the ARI-A resistance island. These findings underlined that IncX3 plasmids have played a major role in the dissemination of blaNDM-like genes in Czech hospitals. In combination with further evolvement of NDM-like-encoding MDR plasmids through reshuffling, NDM-like producers pose an important public threat.

• Publikační typ
• časopisecké články MeSH

The spread of antimicrobial resistance from human activity derived sources to natural habitats implicates wildlife as potential vectors of antimicrobial resistance transfer. Wild birds, including corvid species can disseminate mobile genetic resistance determinants through faeces. This study aimed to determine the occurrence of plasmid-mediated quinolone resistance (PMQR) genes in Escherichia coli and Klebsiella spp. isolates obtained from winter roosting sites of American crows (Corvus brachyrhynchos) and common ravens (Corvus corax) in Canada. Faecal swabs were collected at five roosting sites across Canada. Selective media isolation and multiplex PCR screening was utilized to identify PMQR genes followed by gene sequencing, pulse-field gel electrophoresis and multilocus sequence typing to characterize isolates. Despite the low prevalence of E. coli containing PMQR (1·3%, 6/449), qnrS1, qnrB19, qnrC, oqxAB and aac(6')-Ib-cr genes were found in five sequence types (ST), including E. coli ST 131. Conversely, one isolate of Klebsiella pneumoniae contained the plasmid-mediated resistance gene qnrB19. Five different K. pneumoniae STs were identified, including two novel types. The occurrence of PMQR genes and STs of public health significance in E. coli and Klebsiella pneumoniae recovered from corvids gives further evidence of the anthropogenic derived dissemination of antimicrobial resistance determinants at the human activity-wildlife-environment interface. SIGNIFICANCE AND IMPACT OF THE STUDY: This study examined large corvids as possible vector species for the dissemination of antimicrobial resistance in indicator and pathogenic bacteria as a means to assess the anthropogenic dissemination of plasmid-mediated quinolone resistance (PMQR) genes. Although rare, PMQR genes were found among corvid populations across Canada. The clinically important Escherichia coli strain ST131 containing aac(6')-Ib-cr gene along with a four-class phenotypic antimicrobial resistance (AMR) pattern as well as one Klebsiella pneumoniae strain containing a qnrB19 gene were identified in one geographical location. Corvids are a viable vector for the circulation of PMQR genes and clinically important clones in wide-ranging environments.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence genetika   MeSH
• chinolony farmakologie   MeSH
• Escherichia coli účinky léků   genetika   izolace a purifikace   MeSH
• infekce bakteriemi rodu Klebsiella farmakoterapie   mikrobiologie   MeSH
• infekce vyvolané Escherichia coli farmakoterapie   mikrobiologie   MeSH
• Klebsiella pneumoniae účinky léků   genetika   izolace a purifikace   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• multilokusová sekvenční typizace MeSH
• nemoci ptáků farmakoterapie   mikrobiologie   MeSH
• plazmidy genetika   MeSH
• průřezové studie MeSH
• vrány mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Kanada MeSH

Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (∼30, ∼45, and ∼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.

• MeSH
• antibakteriální látky terapeutické užití   MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• Citrobacter freundii účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• enterobakteriální infekce farmakoterapie   epidemiologie   mikrobiologie   MeSH
• Escherichia coli účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• exprese genu MeSH
• izoenzymy genetika   metabolismus   MeSH
• karbapenemy terapeutické užití   MeSH
• Klebsiella pneumoniae účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• Morganella morganii účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• nemocnice MeSH
• otevřené čtecí rámce MeSH
• plazmidy chemie   klasifikace   metabolismus   MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• transpozibilní elementy DNA MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

A VIM-1-producing ST92 Enterobacter cloacae was isolated in a Czech hospital. blaVIM-1 was part of the class 1 integron In110 carried by a Tn1721-like transposon. Tn1721-like was located on a ColE1-like plasmid, pEncl-30969cz (33,003 bp). Target site duplications at the boundaries of Tn1721-like suggested its transposition into the pEncl-30969cz backbone.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny genetika   MeSH
• beta-laktamasy genetika   MeSH
• beta-laktamová rezistence genetika   MeSH
• DNA bakterií analýza   genetika   MeSH
• Enterobacter cloacae genetika   MeSH
• enterobakteriální infekce mikrobiologie   MeSH
• integrony genetika   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• plazmidy genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články 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