The aim of this study was to fabricate novel microparticles (MPs) for efficient and long-term delivery of amikacin (AMI). The emulsification method proposed for encapsulating AMI employed low-molecular-weight poly(lactic acid) (PLA) and poly(lactic acid-co-polyethylene glycol) (PLA-PEG), both supplemented with poly(vinyl alcohol) (PVA). The diameters of the particles obtained were determined as less than 30 μm. Based on an in-vitro release study, it was proven that the MPs (both PLA/PVA- and PLA-PEG/PVA-based) demonstrated long-term AMI release (2 months), the kinetics of which adhered to the Korsmeyer-Peppas model. The loading efficiencies of AMI in the study were determined at the followings levels: 36.5 ± 1.5 μg/mg for the PLA-based MPs and 106 ± 32 μg/mg for the PLA-PEG-based MPs. These values were relatively high and draw parallels with studies published on the encapsulation of aminoglycosides. The MPs provided antimicrobial action against the Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae bacterial strains. The materials were also comprehensively characterized by the following methods: differential scanning calorimetry; gel permeation chromatography; scanning electron microscopy; Fourier transform infrared spectroscopy-attenuated total reflectance; energy-dispersive X-ray fluorescence; and Brunauer-Emmett-Teller surface area analysis. The findings of this study contribute toward discerning new means for conducting targeted therapy with polar, broad spectrum antibiotics.

• MeSH
• amikacin aplikace a dávkování   chemie   MeSH
• antibakteriální látky aplikace a dávkování   chemie   MeSH
• Escherichia coli účinky léků   MeSH
• Klebsiella pneumoniae účinky léků   MeSH
• laktáty chemie   MeSH
• mikrobiální testy citlivosti MeSH
• molekulová hmotnost MeSH
• nosiče léků chemie   MeSH
• polyestery chemie   MeSH
• polyethylenglykoly chemie   MeSH
• polyvinylalkohol chemie   MeSH
• příprava léků metody   MeSH
• Pseudomonas aeruginosa účinky léků   MeSH
• rozpustnost MeSH
• Staphylococcus aureus účinky léků   MeSH
• tobolky MeSH
• uvolňování léčiv MeSH
• velikost částic MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In the microbiological diagnosis of bloodstream infections (BSI), blood culture (BC) is considered the gold standard test despite its limitations such as low sensitivity and slow turnaround time. A new FDA-cleared and CE-marked platform utilizing magnetic resonance to detect amplified DNA of the six most common and/or problematic BSI pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli; referred to as ESKAPEc) is available and may shorten the time to diagnosis and potentially improve antimicrobial utilization. Whole blood samples from hospitalized patients with clinical signs of sepsis were analyzed using the T2Bacteria Panel (T2Biosystems) and compared to simultaneously collected BC. Discrepant results were evaluated based on clinical infection criteria, combining supporting culture results and the opinion of treating physicians. A total of 55 samples from 53 patients were evaluated. The sensitivity and specificity of the T2Bacteria panel was 94% (16 out of 17 detections of T2Bacteria-targeted organisms) and 100%, respectively, with 36.4% (8 of 22) causes of BSI detected only by this method. The T2Bacteria Panel detected pathogens on average 55 hours faster than standard BC. In our study, 9 of 15 patients with positive T2Bacteria Panel results received early-targeted antibiotic therapy and/or modification of antimicrobial treatment based on T2Bacteria Panel findings. Given the high reliability, faster time to detection, and easy workflow, the technique qualifies as a point-of-care testing approach.

• MeSH
• Acinetobacter baumannii účinky léků   genetika   izolace a purifikace   MeSH
• antibakteriální látky farmakologie   MeSH
• antibiotická politika metody   MeSH
• bakteriemie krev   farmakoterapie   mikrobiologie   MeSH
• Enterococcus faecium účinky léků   genetika   izolace a purifikace   MeSH
• Escherichia coli účinky léků   genetika   izolace a purifikace   MeSH
• Klebsiella pneumoniae účinky léků   genetika   izolace a purifikace   MeSH
• krev mikrobiologie   MeSH
• kultivační vyšetření krve MeSH
• lidé MeSH
• prospektivní studie MeSH
• Pseudomonas aeruginosa účinky léků   genetika   izolace a purifikace   MeSH
• Staphylococcus aureus účinky léků   genetika   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Klebsiella pneumoniae (family Enterobacteriaceae) is a gram-negative bacterium that has strong pathogenicity to humans and can cause sepsis, pneumonia, and urinary tract infection. In recent years, the unreasonable use of antibacterial drugs has led to an increase in drug-resistant strains of K. pneumoniae, a serious threat to public health. Bacteriophages, viruses that infect bacteria, are ubiquitous in the natural environment. They are considered to be the most promising substitute for antibiotics because of their high specificity, high efficiency, high safety, low cost, and short development cycle. In this study, a novel phage designated vB_KpnP_IME279 was successfully isolated from hospital sewage using a multidrug-resistant strain of K. pneumoniae as an indicator. A one-step growth curve showed that vB_KpnP_IME279 has a burst size of 140 plaque-forming units/cell and a latent period of 20 min at its optimal multiplicity of infection (MOI = 0.1). Phage vB_KpnP_IME279 survives in a wide pH range between 3 and 11 and is stable at temperatures ranging from 40 to 60 °C. Ten of the 20 strains of K. pneumoniae including the host bacteria were lysed by the phage vB_KpnP_IME279, and the multilocus sequence typing and wzi typing of the 10 strains were ST11, ST37, ST375, wzi209, wzi52, and wzi72, respectively. The genome of vB_KpnP_IME279 is 42,518 bp long with a G + C content of 59.3%. Electron microscopic observation showed that the phage belongs to the family Podoviridae. BLASTN alignment showed that the genome of the phage has low similarity with currently known phages. The evolutionary relationship between phage vB_KpnP_IME279 and other Podoviridae was analyzed using a phylogenetic tree based on sequences of phage major capsid protein and indicates that the phage vB_KpnP_IME279 belongs to the Podoviridae subfamily. These data enhance understanding of K. pneumoniae phages and will help in development of treatments for multidrug-resistant bacteria using phages.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriofágy klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• fylogeneze MeSH
• genom virový MeSH
• hostitelská specificita MeSH
• Klebsiella pneumoniae účinky léků   izolace a purifikace   virologie   MeSH
• lidé MeSH
• mikrobiologické techniky MeSH
• mnohočetná bakteriální léková rezistence MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• odpadní vody mikrobiologie   virologie   MeSH
• Podoviridae klasifikace   genetika   izolace a purifikace   MeSH
• RNA ribozomální 16S MeSH
• sekvenování celého genomu MeSH
• techniky typizace bakterií MeSH
• teplota MeSH
• zastoupení bazí MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Cíl práce: Tento článek přináší přehled nejčastějších bakteriálních druhů izolovaných z hemokultur ve Fakultní nemocnici Olomouc v letech 2015–2019 a jejich profil rezistence k antibiotikům. Materiál a metody: Data byla získána z laboratorního informačního systému ENVIS LIMS. Za období 1. 1. 2015 až 31. 12. 2019 byly retrospektivně hodnoceny výsledky hemokultivací. Z pozitivních hemokultur byla vyhodnocena četnost bakteriálních izolátů a u nejčastějších původců bakteriémií zjištěna rezistence k vybraným antibiotikům. K identifikaci byly použity standardní mikrobiologické postupy za pomoci systému MALDI-TOF MS. Citlivost k antibiotikům byla stanovena standardní diluční mikrometodou podle kritérií EUCAST. Výsledky: V období 5 let bylo celkem hodnoceno 3 400 izolátů z pozitivních hemokultur. Nejčastěji byly izolovány koaguláza-negativní stafylokoky (37 %), Escherichia coli (16 %), Klebsiella pneumoniae (9 %), Staphylococcus aureus (7 %), streptokoky (5 %), Pseudomonas aeruginosa (4 %), Enterobacter cloacae (2 %), Enterococcus faecalis (2 %) a Enterococcus faecium (2 %). Escherichia coli byla v uvedeném období relativně málo rezistentní k cefalosporinům 3. generace, piperacilinu s tazobaktamem, gentamicinu a fluorochinolonům (7–33 %). Klebsiella pneumoniae naproti tomu vykazovala vyšší rezistenci k těmto antibiotikům (33–65 %). Rezistence druhu Staphylococcus aureus k oxacilinu se pohybuje v rozmezí 3–7 %. Pseudomonas aeruginosa se vyznačovala rezistencí alespoň k jednomu z následujících antibiotik – piperacilinu/tazobaktamu, ceftazidimu, cefepimu, meropenemu, ciprofloxacinu a gentamicinu v rozmezí 3–46 %. Závěr: Znalost prevalence původců bakteriémií a jejich profilů rezistence je důležitá pro iniciální antibiotickou terapii v případě probíhající infekce. Je vhodné epidemiologické přehledy tohoto typu pravidelně provádět a poskytovat klinickým lékařům jako součást „antibiotic stewardship“.

Objectives: To provide an overview of the most common bacterial species isolated from blood cultures in the University Hospital Olomouc in the years 2015–2019 and their antibiotic resistance patterns. Material and Methods: The data were obtained from the laboratory information management system ENVIS LIMS. The results were analyzed retrospectively for the period from January 1, 2015 to December 31, 2019. Among positive blood cultures, the prevalence of bacterial species was assessed and the most frequent species were evaluated for resistance to selected antibiotics. Each sample was processed using standard microbiology methods with the MALDI-TOF MS system. Susceptibility to antibiotics was tested with the microdilution method according to the EUCAST recommendations. Results: Over the study period, a total of 3 400 isolates from blood cultures were included. Coagulase-negative staphylococci were the most prevalent (37 %), followed by Escherichia coli (16 %), Klebsiella pneumoniae (9 %), Staphylococcus aureus (7 %), Streptococcus spp. (5 %), Pseudomonas aeruginosa (4 %), Enterobacter cloacae (2 %), Enterococcus faecalis (2 %) and Enterococcus faecium (2 %). Resistance of E. coli to 3rd generation cephalosporins, piperacillin/tazobactam, gentamicin and fluoroquinolones ranged from 7 % to 33 %. A high percentage of Klebsiella pneumoniae strains (33 %–65 %) was resistant to the above antibiotics. The prevalence of methicillin-resistant Staphylococcus aureus reached 3 %–7 %. Pseudomonas aeruginosa exhibited resistance to piperacillin/tazobactam, ceftazidime, cefepime, meropenem, ciprofloxacin and gentamicin ranging from 3 % to 46 %. Conclusion: Knowing the prevalence of bacterial species from blood cultures and their antimicrobial resistance patterns is important for empirical antibiotic therapy in case of an existing infection. It is advisable to conduct such epidemiological studies as part of antibiotic stewardship.

• MeSH
• antibiotická rezistence * MeSH
• bakteriální léková rezistence MeSH
• bakteriemie * krev   mikrobiologie   terapie   MeSH
• Escherichia coli účinky léků   MeSH
• Klebsiella pneumoniae účinky léků   MeSH
• kultivační vyšetření krve * metody   MeSH
• lidé MeSH
• mikrobiální testy citlivosti metody   MeSH
• mikrobiologické techniky metody   MeSH
• Pseudomonas aeruginosa účinky léků   MeSH
• Staphylococcus aureus účinky léků   MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Česká republika MeSH

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

We investigated the occurrence of plasmid-mediated colistin resistance in Enterobacteriaceae in turkey meat produced in the Czech Republic as well as in turkey meat imported into the Czech Republic from other European Union countries. Seventeen samples of raw turkey meat from the Czech Republic (n = 4), Hungary (n = 2), Poland (n = 6) and Germany (n = 5) were cultured in peptone water at 37 °C overnight and the enriched cultures were tested for the presence of mcr-1-5 genes. PCR-positive enriched cultures were inoculated onto selective agar with colistin (3.5 mg/L). A minimal inhibitory concentration (MIC) of colistin was determined by using the broth microdilution method in PCR-positive isolates. In addition, a macrorestriction analysis was performed using XbaI endonuclease. Of 17 meat samples, 12 samples from Poland (6/6), Germany (3/5) and the Czech Republic (3/4) proved positive for the presence of the mcr-1 gene. Forty-two isolates carrying the mcr-1 gene were obtained: Escherichia coli (n = 39) revealing 32 distinct XbaI profiles and Klebsiella pneumoniae (n = 3) with 2 distinct XbaI profiles. The minimal inhibitory concentration (MIC) of the mcr-1 positive isolates was as follows: 4 mg/L (n = 28), 8 mg/L (n = 12), 32 mg/L (n = 1) and 64 mg/L (n = 1). The high prevalence (70.6%; 12/17 samples) of mcr-1-mediated colistin-resistant Enterobacteriaceae found in the turkey meat samples analysed in this study, builds on previously published evidence that poultry, and their products, represent a substantial risk for the dissemination of plasmid-mediated colistin resistance in Europe.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence genetika   MeSH
• drůbeží výrobky mikrobiologie   MeSH
• Escherichia coli účinky léků   genetika   MeSH
• Klebsiella pneumoniae účinky léků   genetika   MeSH
• kolistin farmakologie   MeSH
• krocani mikrobiologie   MeSH
• mikrobiální testy citlivosti MeSH
• plazmidy genetika   MeSH
• proteiny z Escherichia coli genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika 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

BackgroundAntibiotic resistance, either intrinsic or acquired, is a major obstacle for treating bacterial infections.AimOur objective was to compare the country-specific species distribution of the four Gram-negative species Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter species and the proportions of selected acquired resistance traits within these species.MethodWe used data reported for 2016 to the European Antimicrobial Resistance Surveillance Network (EARS-Net) by 30 countries in the European Union and European Economic Area.ResultsThe country-specific species distribution varied considerably. While E. coli accounted for 31.9% to 81.0% (median: 69.0%) of all reported isolates, the two most common intrinsically resistant species P. aeruginosa and Acinetobacter spp. combined (PSEACI) accounted for 5.5% to 39.2% of isolates (median: 10.1%). Similarly, large national differences were noted for the percentages of acquired non-susceptibility to third-generation cephalosporins, carbapenems and fluoroquinolones. There was a strong positive rank correlation between the country-specific percentages of PSEACI and the percentages of non-susceptibility to the above antibiotics in all four species (rho > 0.75 for 10 of the 11 pairs of variables tested).ConclusionCountries with the highest proportion of P. aeruginosa and Acinetobacter spp. were also those where the rates of acquired non-susceptibility in all four studied species were highest. The differences are probably related to national differences in antibiotic consumption and infection prevention and control routines.

• MeSH
• Acinetobacter účinky léků   MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence účinky léků   MeSH
• bakteriemie farmakoterapie   epidemiologie   MeSH
• cefalosporiny farmakologie   MeSH
• Escherichia coli účinky léků   MeSH
• Evropská unie MeSH
• fluorochinolony farmakologie   MeSH
• gramnegativní bakteriální infekce farmakoterapie   epidemiologie   MeSH
• gramnegativní bakterie účinky léků   MeSH
• karbapenemy farmakologie   MeSH
• Klebsiella pneumoniae účinky léků   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• Pseudomonas aeruginosa účinky léků   MeSH
• sentinelová surveillance MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

Diferenciální diagnostika horečky u kojenců je široká. Po vyloučení infekce respirační, močové a gastrointestinální je nutno pomýšlet i na neuroinfekce. Prezentované kazuistické sdělení se věnuje časné diagnostice infekce centrální nervové soustavy a problematice interpretace zánětlivých parametrů v časné fázi bakteriálního infektu. Zdůrazňuje fakt, že časný likvorový nález u bakteriální meningitidy nemusí být zcela rozvinut a upozorňuje na obtíže při interpretaci zánětlivých parametrů ve vztahu k jejich biologickému poločasu.

In infant period, the differential diagnostics of fever is large. After eliminating respiratory, urinary and gastrointestinal infection, neuroinfection should also be taken into account. The presented case report deals with early diagnostics of central nervous system infection and with the issue of interpretation of inflammatory parameters in the early stage of a bacterial infection. It emphasises the fact that the early cerebrospinal fluid findings in the case of bacterial meningitis may not always be developed and calls attention to difficulties in the interpretation of inflammatory parameters with regard to their biological half-time.

• Klíčová slova
• zánětlivé parametry,
• MeSH
• ceftriaxon terapeutické užití   MeSH
• diagnostické techniky a postupy MeSH
• infekce centrálního nervového systému * diagnóza   terapie   MeSH
• Klebsiella pneumoniae účinky léků   MeSH
• klinické laboratorní techniky MeSH
• lidé MeSH
• meningitida * diagnóza   terapie   MeSH
• novorozenec MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• Publikační typ
• kazuistiky MeSH

BACKGROUND: Bacteraemia of the donor is not considered to be contraindication of organ procurement. On the other hand, infection of solid organ transplant recipients remains to be a major cause of their morbidity and mortality. When using organs from bacteraemic donors, individual risks need to be assessed and the appropriate antibiotic treatment applied. CASE PRESENTATION: In this case series we report several serious donor-derived infectious complications in four out of five recipients of different organs from one single donor in the early posttransplant period. Donor-transmitted multi-drug resistant strains of Escherichia coli and Klebsiella pneumonia was confirmed by both serologic and molecular testing. CONCLUSIONS: To prevent donor-derived infections, careful microbiological screening followed by targeted antibiotic treatment is essential. Although such complications can never by completely prevented, a high index for potential bacterial infection in organ donors and transplant recipients should be routinely employed.

• MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriemie * MeSH
• dárci tkání * MeSH
• dospělí MeSH
• Escherichia coli účinky léků   MeSH
• infekce bakteriemi rodu Klebsiella diagnóza   farmakoterapie   mikrobiologie   MeSH
• infekce vyvolané Escherichia coli diagnóza   farmakoterapie   mikrobiologie   MeSH
• Klebsiella pneumoniae účinky léků   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetná bakteriální léková rezistence * MeSH
• příjemce transplantátu MeSH
• transplantace orgánů škodlivé účinky   MeSH
• získávání tkání a orgánů MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH