The occurrence of antimicrobial-resistant bacteria is an important public health issue. The aim of this study was the monitoring of resistant Escherichia coli in raw cow's milk with a focus on the detection of extended-spectrum β-lactamase (ESBL)-producing strains. In total, 263 samples of raw milk from 40 farms were collected and investigated in 2010 to 2013 in the Czech Republic. Detection of E. coli was performed and evaluated according to ISO 16649-2, and antibiotic resistance was screened by the disk diffusion method. The presence of E. coli was detected in 243 (92.4%) samples. In total, 270 isolates were obtained. Resistance to β-lactam (31.8%) and tetracycline (13.0%) antibiotics was detected most often and also multiresistant strains (5.5%) were observed. E. coli isolates found to be resistant to β-lactam, tetracycline, and quinolone antibiotics were assayed by PCR to detect selected genes encoding those resistance mechanisms. In isolates in which any bla genes were detected, a double-disk synergy test was performed. ESBL production was confirmed in 2 (0.7%) isolates. The genetic analysis identified the presence of the blaCTX-M gene and other resistance genes (tet(B) and qnrB). Both ESBL-positive isolates originated from the same farm and had an identical pulsed-field gel electrophoresis profile. The findings of our study indicate that milk can be a reservoir of bacteria carrying resistance genes with a potential for spreading through the food chain.

• MeSH
• antibakteriální látky farmakologie   MeSH
• beta-laktamasy biosyntéza   genetika   MeSH
• Escherichia coli účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• mléko mikrobiologie   MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• polymerázová řetězová reakce MeSH
• proteiny z Escherichia coli genetika   MeSH
• pulzní gelová elektroforéza MeSH
• skot MeSH
• syrová strava mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

The entire plasmid content of a multidrug-resistant, CTX-M-15-producing Klebsiella pneumoniae ST416 clone was investigated. Two FII(K) plasmids, pKDO1 (127 kb) and pKPN-CZ (207 kb), were identified and found to carry a formidable set of genes conferring resistance to toxic compounds, metals, and antimicrobial drugs and exhibiting novel features putatively associated with adaptation and fitness of the bacterium in the human host.

• MeSH
• antiinfekční látky farmakologie   MeSH
• bakteriální proteiny biosyntéza   metabolismus   MeSH
• beta-laktamasy biosyntéza   metabolismus   MeSH
• infekce bakteriemi rodu Klebsiella farmakoterapie   MeSH
• Klebsiella pneumoniae účinky léků   genetika   MeSH
• kovy farmakologie   MeSH
• mikrobiální testy citlivosti MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• molekulární sekvence - údaje MeSH
• plazmidy genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• antibakteriální látky farmakologie   MeSH
• beta-laktamasy biosyntéza   genetika   MeSH
• beta-laktamová rezistence genetika   MeSH
• infekce spojené se zdravotní péčí epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice fakultní MeSH
• polymerázová řetězová reakce metody   MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   MeSH
• Pseudomonas genetika   klasifikace   účinky léků   MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Česká republika MeSH

Resistance of bacteria to antibiotics is a global medical problem requiring close cooperation between veterinary and human physicians. Raw materials and foods of animal origin may be not only a source of pathogenic bacteria causing alimentary tract infections but also a source of bacteria with a dangerous extent of resistance to antibiotics, potentially entering the human food chain. This article presents results of the first study in the Czech Republic detecting the presence of Enterobacteriaceae-producing extended-spectrum b -lactamases (ESBLs) in swabs collected in slaughterhouses from surfaces of healthy animal carcasses. In 2012, swabs taken from pig (n = 166) and cattle (n = 140) carcass surfaces were analyzed. In 17 % of 53 studied slaughterhouses, ESBL-producing Escherichia coli strains were isolated. ESBLs were found in 11 and 4 % of porcine and bovine samples, respectively. Swabs collected from pigs yielded 18 ESBL-producing E. coli strains. The bla genes were found to encode production of CTX-M-1 group enzymes in 16 strains, SHV in one case, and both CTX-M-1-like and TEM in another case. In swabs taken from cattle, five ESBL-producing E. coli strains were isolated. In three cases, the bla genes for CTX-M-1-like production were identified; in two cases, genes for both CTX-M-1-like and TEM production were found. The similarity/identity of ESBL-positive isolates was compared by pulsed-field gel electrophoresis. This is the first report and characterization of the presence and nature of ESBL-producing E. coli in swabs collected from surfaces of healthy pig and cattle carcasses in slaughterhouses in the Czech Republic.

• MeSH
• beta-laktamasy biosyntéza   genetika   izolace a purifikace   MeSH
• Escherichia coli enzymologie   genetika   izolace a purifikace   MeSH
• jatka * MeSH
• lidé MeSH
• potravinářská mikrobiologie MeSH
• prasata mikrobiologie   MeSH
• prevalence MeSH
• pulzní gelová elektroforéza MeSH
• skot mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot mikrobiologie   MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

AIM: The study aimed at analyzing ESBL- and AmpC-positive Enterobacteriaceae in the gastrointestinal tracts of university hospital inpatients and persons from the Olomouc Region community, and comparing the results with data from 2007. METHODS: Bacteria were isolated from rectal swabs inoculated onto the ChromID(TM) ESBL selective medium (bioMérieux). Production of ESBL-type beta-lactamases was confirmed by the modified double-disk synergy test and AmpC enzyme production was detected by the AmpC disk test. ESBL- and AmpC-positive isolates were subjected to basic genetic analysis aimed at detecting the bla(TEM), bla(SHV), bla(CTX-M) and bla(AmpC) genes. RESULTS: Over the study period (1 March 2010 - 1 May 2010), a total of 1,279 rectal swabs (70.4% of community subjects) were analyzed on the above medium. The prevalence rates of ESBL-positive Enterobacteriaceae were 8.2% in hospitalized patients and 3.2% in community subjects. Production of the AmpC enzyme was detected in 1.1% of bacterial isolates from the community and in one (0.3%) hospital isolate. Among ESBL, the most frequent genes encoding enzymes were from the CTX-M-1-like genes. Detected AmpC beta-lactamases belonged to the CIT, DHA and EBC groups. CONCLUSION: When compared with the year 2007, the rates of carriers of ESBL-positive bacteria increased in both hospitalized patients (from 3% to 8%) and community subjects (from 1% to 3%) in 2010. Given the fact that production of extended-spectrum beta-lactamases is clinically significant, knowing the epidemiological situation is very important for selecting adequate antibiotic therapy.

• MeSH
• bakteriální proteiny biosyntéza   MeSH
• beta-laktamasy biosyntéza   MeSH
• Enterobacteriaceae enzymologie   izolace a purifikace   MeSH
• gastrointestinální trakt mikrobiologie   MeSH
• lidé MeSH
• pacienti hospitalizovaní MeSH
• přenašečství MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

We describe the isolation of an NDM-1-producing Acinetobacter baumannii in a Czech patient repatriated in July 2011 from Egypt. The infection spread to another patient on the same ward. Both isolates showed the same resistance pattern and were susceptible only to colistin. They had an identical PFGE pattern and belonged to the same sequence type ST 1. Sequencing of the blaNDM gene identified the NDM-1 variant of the carbapenemase, surrounded by two copies of insertion sequence ISAba125.

• MeSH
• Acinetobacter baumannii enzymologie   genetika   izolace a purifikace   MeSH
• amplifikace genu MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriální proteiny biosyntéza   MeSH
• beta-laktamasy biosyntéza   genetika   MeSH
• beta-laktamová rezistence MeSH
• cestování MeSH
• infekce bakteriemi rodu Acinetobacter diagnóza   farmakoterapie   přenos   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• pulzní gelová elektroforéza MeSH
• techniky typizace bakterií MeSH
• ventilátorová pneumonie mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Egypt MeSH

Carbapenemase-producing Gram-negative bacteria peak clinical interest due to their ability to hydrolyze most β-lactams, including carbapenems; moreover, their genes spread through bacterial populations by horizontal transfer. Bacteria with acquired carbapenemase have sporadically been reported in the Czech Republic, so far only in Enterobacteriaceae and Pseudomonas aeruginosa. In this study, we described the first finding of a KPC-2-producing strain of Klebsiella pneumoniae, which was isolated from a surgical wound swab, decubitus ulcer, and urine of a patient previously hospitalized in Greece. The patient underwent various antibiotic therapies including a colistin treatment. However, after approximately 20 days of the colistin therapy, the strain developed a high-level resistance to this drug. All the isolates were indistinguishable by pulsed field gel electrophoretic analysis and belonged to the international clone ST258, which is typical of KPC-producing K. pneumoniae isolates. The bla (KPC-2) gene was located on a Tn4401a transposon variant. The OmpK35 and OmpK36 genes analysis performed due to the high resistance level of the strains to β-lactams exhibited no changes in their sequence or in their expression when compared with carbapenem-susceptible isolates.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence účinky léků   genetika   MeSH
• beta-laktamasy biosyntéza   genetika   MeSH
• hospitalizace MeSH
• Klebsiella pneumoniae účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• kolistin farmakologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• poriny genetika   MeSH
• regulace genové exprese u bakterií MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Řecko MeSH

O interpretaci citlivosti k beta-laktamovým antibiotikům u gramnegativních tyček produkujících beta-laktamázy se v současnosti diskutuje v CLSI i v EUCAST, renomovaných institucích doporučujících klinické break-pointy. Článek shrnuje současné poznatky o break-pointech u enterobakterií a navrhuje postup pro klinicko-mikrobiologické laboratoře České republiky.

The interpretation of the susceptibility of Gram-negative rods to ß-lactams is currently under discussion in CLSI and EUCAST – two authorities on determination of clinical breakpoints. This article summarizes the current knowledge about clinical breakpoints in enterobacteria and proposes guidance for clinical microbiology laboratories in the Czech Republic.

• Klíčová slova
• MBL, KPC, karbapenemázy, AmpC, ESBL,
• MeSH
• beta-laktamasy biosyntéza   MeSH
• beta-laktamy farmakokinetika   farmakologie   MeSH
• cefalosporiny farmakokinetika   farmakologie   MeSH
• Enterobacteriaceae enzymologie   metabolismus   účinky záření   MeSH
• hydrolýza MeSH
• karbapenemy farmakokinetika   farmakologie   MeSH
• mikrobiální testy citlivosti MeSH

Individual cloacal swabs of mallards (Anas platyrhynchos) and of herring gulls (Larus argentatus), as well as samples of waterbird feces obtained in 2008 and 2009, were cultivated for Escherichia coli. Isolates of E. coli were tested for susceptibilities to 12 antimicrobial agents by the disk diffusion method. Moreover, the samples were subcultivated on MacConkey agar (MCA) containing cefotaxime (2 mg liter(-1)) to detect E. coli with extended-spectrum beta-lactamase (ESBL) and subsequently on MCA supplemented with ciprofloxacin (0.05 mg liter(-1)) and MCA with nalidixic acid (20 mg liter(-1)) to isolate fluoroquinolone-resistant E. coli. PCR was used to detect specific antibiotic resistance genes. We found 9 E. coli isolates producing ESBL with bla genes: bla(CTX-M-1) (6 isolates), bla(CTX-M-9) plus bla(TEM-1b) (1 isolate), bla(CTX-M-15) plus bla(OXA-1) (1 isolate), and bla(SHV-12) (1 isolate). In the isolate with bla(CTX-M-15), the gene aac(6)-Ib-cr was also detected. The bla genes were harbored by transferable plasmids of the IncN and IncI1 groups. Nine quinolone-resistant E. coli isolates with qnrS genes were found and characterized. The gene qnrS was associated with a Tn3-like transposon on the IncX1 plasmid together with bla(TEM-1) in two isolates. The gene qnrS was also harbored by conjugative plasmids of the IncN and IncX2 groups. Even if populations of wild birds are not directly influenced by antibiotic practice, we have demonstrated that antibiotic-resistant E. coli strains, including strains with various ESBL and qnrS genes, are found in the feces of wild birds on the coast of the Baltic Sea in Poland.

• MeSH
• Anseriformes mikrobiologie   MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální geny MeSH
• bakteriální léková rezistence MeSH
• beta-laktamasy biosyntéza   MeSH
• Charadriiformes mikrobiologie   MeSH
• Escherichia coli účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• feces mikrobiologie   MeSH
• kloaka mikrobiologie   MeSH
• kultivační média chemie   MeSH
• mikrobiální testy citlivosti MeSH
• oceány a moře MeSH
• plazmidy analýza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• oceány a moře MeSH
• Polsko MeSH

Thirty-eight AmpC-producing Klebsiella pneumoniae isolates identified from January to October 2006 in a large teaching hospital in the Czech Republic were analyzed. The AmpC cephalosporinase was identified as DHA-1, encoded by a plasmid-located complex class 1 integron, previously observed in a K. pneumoniae isolate from the Parisian region. The DHA-1 expression was inducible, and although in two isolates with higher resistance, the induction effect was masked at the phenotypic level. All of the isolates belonged to the international K. pneumoniae clone sequence type 11, split into two disseminated pulsed-field gel electrophoresis types. This is the first report on enterobacteriaceae with acquired AmpCs in the Czech Republic and possibly the first description of organisms with DHA-1 in the Central and Eastern Europe.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny biosyntéza   genetika   MeSH
• beta-laktamasy biosyntéza   genetika   MeSH
• infekce bakteriemi rodu Klebsiella epidemiologie   mikrobiologie   MeSH
• integrony genetika   MeSH
• Klebsiella pneumoniae účinky léků   enzymologie   genetika   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• nemocnice fakultní * MeSH
• plazmidy genetika   MeSH
• pulzní gelová elektroforéza MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH