Bakterie komplexu Burkholderia cepacia vyvolávají u pacientů s cystickou fibrózou (CF) závažné plicní infekce. Riziko epidemického šíření infekce přimělo centra CF zavést přísná izolační opatření. Navíc je nutné správně a včas mikrobiálního původce identifikovat, k čemuž slouží postupy molekulární mikrobiologie. V posledním desetiletí nebyl v ČR diagnostikován žádný nový případ infekce způsobené epidemickým kmenem ST-32 druhu Burkholderia cenocepacia, který stál za velkou epidemií 90. let 20. století.
Bacteria from the Burkholderia cepacia complex cause severe lung infections in people with cystic fibrosis (CF). The risk of epidemic spread of the infection led the CF centres to implement strict isolation precautions. Furthermore, it was imperative to identify the microbial agent correctly and timely which was made possible thanks to the methods of molecular microbiology. No new Burkholderia cenocepacia infections, caused by the ST-32 strain which had been responsible for a large outbreak in 1990s, were diagnosed in the Czech Republic in the last ten years.
Bacteria from the Burkholderia cepacia complex are generally considered to be non-pathogenic to the healthy population. However, some of these species may cause serious nosocomial infections in immunocompromised patients; as such, it is essential to diagnose these infections rapidly so that adequate treatment can be initiated. We report here the use of a radiolabeled siderophore, ornibactin (ORNB), for positron emission tomography imaging. We successfully radiolabeled ORNB with gallium-68 with high radiochemical purity and proved that the resulting complex has optimal in vitro characteristics. In mice, the complex did not show excessive accumulation in organs and was excreted in the urine. We demonstrated that the [68Ga]Ga-ORNB complex accumulates at the site of Burkholderia multivorans infection, including pneumonia, in two animal infection models. These results suggest that [68Ga]Ga-ORNB is a promising tool for the diagnosis, monitoring, and evaluation of the therapeutic response to B. cepacia complex infection.
- MeSH
- Burkholderia cepacia komplex * MeSH
- infekce bakteriemi rodu Burkholderia * diagnostické zobrazování epidemiologie MeSH
- myši MeSH
- pozitronová emisní tomografie MeSH
- radioizotopy galia MeSH
- siderofory MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Polyhydroxyalkanoates (PHAs) are polyesters of significant interest due to their biodegradability and properties similar to petroleum-derived plastics, as well as the fact that they can be produced from renewable sources such as by-product streams. In this study, brewer's spent grain (BSG), the main by-product of the brewing industry, was subjected to a set of physicochemical pretreatments and their effect on the release of reducing sugars (RS) was evaluated. The RS obtained were used as a substrate for further PHA production in Burkholderia cepacia, Bacillus cereus, and Cupriavidus necator in liquid cultures. Although some pretreatments proved efficient in releasing RS (acid-thermal pretreatment up to 42.1 gRS L-1 and 0.77 gRS g-1 dried BSG), the generation of inhibitors in such scenarios likely affected PHA production compared with the process run without pretreatment (direct enzymatic hydrolysis of BSG). Thus, the maximum PHA accumulation from BSG hydrolysates was found in the reference case with 0.31 ± 0.02 g PHA per g cell dried weight, corresponding to 1.13 ± 0.06 g L-1 and a PHA yield of 23 ± 1 mg g-1 BSG. It was also found that C. necator presented the highest PHA accumulation of the tested strains followed closely by B. cepacia, reaching their maxima at 48 h. Although BSG has been used as a source for other bioproducts, these results show the potential of this by-product as a no-cost raw material for producing PHAs in a waste valorization and circular economy scheme.
- Klíčová slova
- ceftazidim/avibactam,
- MeSH
- antibakteriální látky MeSH
- Burkholderia cepacia MeSH
- ceftazidim MeSH
- cystická fibróza * terapie MeSH
- dospělí MeSH
- léková rezistence MeSH
- lidé MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- antibiotická rezistence MeSH
- Burkholderia cepacia komplex izolace a purifikace MeSH
- Corynebacterium izolace a purifikace MeSH
- mikrobiální testy citlivosti metody MeSH
- Salmonella enterica izolace a purifikace MeSH
- Serratia marcescens izolace a purifikace MeSH
- shiga-toxigenní Escherichia coli izolace a purifikace MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- testování odbornosti laboratoří * MeSH
BACKGROUND: Monitoring changes in the epidemiology of cystic fibrosis (CF) pathogens is essential for clinical research, quality improvement, and clinical management. METHODS: We analyzed data reported to the European Cystic Fibrosis Society Patient Registry (ECFSPR) from 2011 to 2016 to determine the overall and the age-specific annual prevalence and incidence of selected CF pathogens and their trends during these years. The ECFSPR collects data on three chronic infections: Pseudomonas aeruginosa (PsA), Burkholderia cepacia complex Species (BCC) and Staphylococcus aureus (SA), as well as on the occurrence of non-tuberculous mycobacteria (NTM) and Stenotrophomonas maltophilia (SM). The same analyses were performed for different country groups, according to their gross national income (GNI). RESULTS: The pathogens with the highest prevalence were SA and PsA, with prevalence, in 2016, equal to 38.3% and 29.8% respectively, followed by SM (8.1%). The pathogens with the lowest prevalence were NTM (3.3%) and BCC (3.1%). The overall prevalence and incidence significantly decreased for PsA; they also decreased for BCC, while they increased significantly for SA. The overall prevalence of NTM and SM increased significantly. The most considerable prevalence changes were observed for PsA, which decreased across all income country groups and all age strata (with the exception of 0-1 years) The prevalence and incidence of pathogens differed significantly according to GNI. CONCLUSIONS: The epidemiology of CF pathogens in Europe has changed; epidemiologic data differ significantly among countries with different socio-economic status. The causes of these observations are multifactorial and include improvements in clinical care and infection control.
- MeSH
- Burkholderia cepacia komplex izolace a purifikace MeSH
- cystická fibróza * epidemiologie mikrobiologie patofyziologie MeSH
- dítě MeSH
- dospělí MeSH
- epidemiologické monitorování MeSH
- infekce dýchací soustavy * epidemiologie mikrobiologie patofyziologie MeSH
- lidé MeSH
- netuberkulózní mykobakterie izolace a purifikace MeSH
- prevalence MeSH
- Pseudomonas aeruginosa izolace a purifikace MeSH
- registrace statistika a číselné údaje MeSH
- socioekonomické faktory MeSH
- Staphylococcus aureus izolace a purifikace MeSH
- Stenotrophomonas maltophilia izolace a purifikace MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
The most problematic respiratory pathogens in patients with cystic fibrosis (CF) are arguably bacteria from Burkholderia cepacia complex (Bcc) as they cause fatal infections, untreatable by current antibiotics. Recently, first promising compounds with a therapeutic potential against Bcc have been discovered, yet underlying molecular processes of their interaction with bacterial cells have not been addressed. The project objectives are to describe inhibitory effect of novel compounds on the transcriptomic level (molecules 11026103 and 10126109 and combination of lactoferrin with hypothiocyanite), to identify targets for 11026103 and 10126109, in relation to variable susceptibility to characterize genome changes introduced over the course of infections with Bcc strain ST32 (dominant among Czech CF patients) and to identify a genotype of resistant ST32 clones. The whole genome sequencing and RNA-seq will be utilized for comparative genomics of ca. 50 longitudinal CF isolates and to describe cellular processes in association with exposure to novel antimicrobial compounds, respectively.
Mezi nejnebezpečnější respirační patogeny patří u pacientů s cystickou fibrózou (CF) bakterie z komplexu Burkholderia cepacia (Bcc), které způsobují fatální infekce, současnými antibiotickými přípravky neléčitelné. První nadějné látky s terapeutickým potenciálem proti Bcc byly nedávno objeveny, ale molekulární procesy blíže určující interakci těchto látek s bakteriální buňkou zůstávají neznámé. Předmětem projektu je popsat na úrovni transkriptomu inhibiční efekt nových antimikrobních preparátů (molekuly 11026103 a 10126109 a kombinace laktoferrinu s hypothiokyanátem), nalézt cílovou strukturu pro 11026103 a 10126109, ve vztahu k rozdílům v citlivosti vůči 4 novým látkám charakterizovat změny v genomu vzniklé v průběhu dlouhodobé infekce u kmene Bcc ST32, který je mezi pacienty s CF v ČR nejzastoupenější, a určit případný genotyp rezistentních klonů ST32. Metody celogenomového sekvenování a RNA-Seq budou užity k porovnání genomů cca 50 longitudinálních izolátů CF a k popisu buněčných procesů odehrávajících se v souvislosti s expozicí novým antimikrobním preparátům.
- MeSH
- antibakteriální látky terapeutické užití MeSH
- Burkholderia cenocepacia MeSH
- cystická fibróza MeSH
- genom MeSH
- infekce bakteriemi rodu Burkholderia terapie MeSH
- sekvenční analýza MeSH
- transkriptom MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- genetika, lékařská genetika
- bakteriologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
Burkholderia cenocepacia causes severe pulmonary infections in cystic fibrosis (CF) patients. Since the bacterium is virtually untreatable by antibiotics, chronic infections persist for years and might develop into fatal septic pneumonia (cepacia syndrome, CS). To devise new strategies to combat chronic B. cenocepacia infections, it is essential to obtain comprehensive knowledge about their pathogenesis. We conducted a comparative genomic analysis of 32 Czech isolates of epidemic clone B. cenocepacia ST32 isolated from various stages of chronic infection in 8 CF patients. High numbers of large-scale deletions were found to occur during chronic infection, affecting preferentially genomic islands and nonessential replicons. Recombination between insertion sequences (IS) was inferred as the mechanism behind deletion formation; the most numerous IS group was specific for the ST32 clone and has undergone transposition burst since its divergence. Genes functionally related to transition metal metabolism were identified as hotspots for deletions and IS insertions. This functional category was also represented among genes where nonsynonymous point mutations and indels occurred parallelly among patients. Another category exhibiting parallel mutations was oxidative stress protection; mutations in catalase KatG resulted in impaired detoxification of hydrogen peroxide. Deep sequencing revealed substantial polymorphism in genes of both categories within the sputum B. cenocepacia ST32 populations, indicating extensive adaptive evolution. Neither oxidative stress response nor transition metal metabolism genes were previously reported to undergo parallel evolution during chronic CF infection. Mutations in katG and copper metabolism genes were overrepresented in patients where chronic infection developed into CS. Among professional phagocytes, macrophages use both hydrogen peroxide and copper for their bactericidal activity; our results thus tentatively point to macrophages as suspects in pathogenesis towards the fatal CS.
- MeSH
- Burkholderia cenocepacia genetika MeSH
- chronická nemoc MeSH
- cystická fibróza komplikace mikrobiologie MeSH
- infekce bakteriemi rodu Burkholderia genetika MeSH
- infekce dýchací soustavy mikrobiologie MeSH
- lidé MeSH
- srovnávací genomová hybridizace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The opportunistic Gram-negative bacterium Burkholderia cenocepacia causes lethal infections in cystic fibrosis patients. Multivalent mannoside derivatives were prepared as potential inhibitors of lectin BC2L-A, one of the virulence factors deployed by B. cenocepacia in the infection process. An (α1→2)-thio-linked mannobioside mimic bearing an azide functionalized aglycon was conjugated to different multivalent scaffolds such as propargylated calix[4]arenes, methyl gallate and pentaerythritol by azide-alkyne 1,3-dipolar cycloaddition. The interaction between the glycoclusters and the mannose binding BC2L-A lectin from B. cenocepacia was examined by isothermal microcalorimetry, surface plasmon resonance, inhibition of yeast agglutination and analytical ultracentrifugation.
- MeSH
- aglutinační testy MeSH
- Burkholderia cenocepacia chemie MeSH
- kalorimetrie metody MeSH
- kvasinky účinky léků MeSH
- lektin vázající mannosu chemie metabolismus farmakologie MeSH
- ligandy MeSH
- mannosidy chemická syntéza chemie metabolismus MeSH
- povrchová plasmonová rezonance MeSH
- techniky syntetické chemie MeSH
- ultracentrifugace metody MeSH
- Publikační typ
- časopisecké články MeSH
