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occurrence of multiple genomovars of burkholderia cepacia in cystic fibrosis patients and proposal of burkholderia multivorans sp. nov.we performed an integrated genotypic and phenotypic analysis of 128 strains of the genera burkholderia, ralstonia, and pseudomonas in order to study the taxonomic structure of burkholderia cepacia and its relationships with other burkholderia species. our data show that presumed b. cepacia strains isolated from cystic fibrosis patients belong to at least five distinct genomic species, one of which was identified as burkholderia vietnamiensis. this group of five phenotypically similar species is ...19979336927
discrimination of burkholderia multivorans and burkholderia vietnamiensis from burkholderia cepacia genomovars i, iii, and iv by pcr.we present a pcr procedure for identification of burkholderia cepacia, burkholderia multivorans, and burkholderia vietnamiensis. 16s and 23s ribosomal dnas (rdnas) of b. multivorans and b. vietnamiensis were sequenced and aligned with published sequences for definition of species-specific 18-mer oligonucleotide primers. specific antisense 16s rdna primers (for b. cepacia, 5'-agc act ccc rcc tct cag-3'; for b. multivorans, 5'-agc act ccc gaa tct ctt-3') and 23s rdna primers (for b. vietnamiensis, ...199910203482
development of rrna-based pcr assays for identification of burkholderia cepacia complex isolates recovered from cystic fibrosis patients.pcr assays targeting rrna genes were developed to identify species (genomovars) within the burkholderia cepacia complex. each assay was tested with 177 bacterial isolates that also underwent taxonomic analysis by whole-cell protein profile. these isolates were from clinical and environmental sources and included 107 b. cepacia complex strains, 23 burkholderia gladioli strains, 20 ralstonia pickettii strains, 10 pseudomonas aeruginosa strains, 8 stenotrophomonas maltophilia strains, and 9 isolate ...199910488171
diagnostically and experimentally useful panel of strains from the burkholderia cepacia complex.two new species, burkholderia multivorans and burkholderia vietnamiensis, and three genomovars (genomovars i, iii, and iv) currently constitute the burkholderia cepacia complex. a panel of 30 well-characterized strains representative of each genomovar and new species was assembled to assist with identification, epidemiological analysis, and virulence studies on this important group of opportunistic pathogens.200010655415
dna-based diagnostic approaches for identification of burkholderia cepacia complex, burkholderia vietnamiensis, burkholderia multivorans, burkholderia stabilis, and burkholderia cepacia genomovars i and iii.bacteria of the burkholderia cepacia complex consist of five discrete genomic species, including genomovars i and iii and three new species: burkholderia multivorans (formerly genomovar ii), burkholderia stabilis (formerly genomovar iv), and burkholderia vietnamiensis (formerly genomovar v). strains of all five genomovars are capable of causing opportunistic human infection, and microbiological identification of these closely related species is difficult. the 16s rrna gene (16s rdna) and reca ge ...200010970351
phenotypic methods for determining genomovar status of the burkholderia cepacia complex.recent taxonomic advances have demonstrated that burkholderia cepacia is a cluster of at least seven closely related genomic species (or genomovars) collectively referred to as the b. cepacia complex, all of which may cause infections among cystic fibrosis patients and other vulnerable individuals. thus, it is important for clinical microbiologists to be able to differentiate genomovars. prior to this study, 361 b. cepacia complex isolates and 51 isolates easily confused with b. cepacia complex ...200111230429
burkholderia cepacia genomovar vi, a new member of the burkholderia cepacia complex isolated from cystic fibrosis patients.a polyphasic taxonomic study was performed on 23 strains isolated from cystic fibrosis (cf) patients in the usa. these strains were tentatively identified as burkholderia cepacia, burkholderia vietnamiensis and burkholderia or ralstonia sp. using biochemical tests and 16s rdna-based pcr assays. visual comparison of protein profiles indicated that they belonged to a single new group ('group 13'). the polyphasic taxonomic data showed that 18 of these strains represent a new member of the b. cepaci ...200111321071
burkholderia cepacia complex: distribution of genomovars among isolates from the maize rhizosphere in italy.burkholderia cepacia is a 'complex' in which seven genomic species or genomovars have so far been identified. it appears that all seven b. cepacia genomovars are capable of causing infections in vulnerable persons; in particular, the importance of burkholderia multivorans (genomovar ii) and b. cepacia genomovar iii among cystic fibrosis isolates, especially epidemic ones, has been emphasized. in order to acquire a better comprehension of the genomovar composition of environmental populations of ...200111321544
[burkholderia cepacia: dangers of a phytopathogen organism for patients with cystic fibrosis].burkholderia cepacia is an environmental bacterium, capable of colonising vegetal and animal tissues, involved in human opportunist nosocomial infections, and above all, in pulmonary colonisations in patients with cystic fibrosis. in these patients, infection may be followed by a severe deterioration with bacteraemia, leading to death. moreover, owing to the epidemic spread of some clones within cystic fibrosis communities, strict preventive guidelines have to be instituted. early detection of b ...200111397673
burkholderia ambifaria sp. nov., a novel member of the burkholderia cepacia complex including biocontrol and cystic fibrosis-related isolates.a polyphasic taxonomic study, including amplified fragment length polymorphism (aflp) fingerprinting, dna-dna hybridizations, dna base-ratio determinations, phylogenetic analysis, whole-cell fatty acid analyses and an extensive biochemical characterization, was performed on 19 burkholderia cepacia-like isolates from the environment and cystic fibrosis (cf) patients. several of the environmental isolates have attracted considerable interest due to their biocontrol properties. the polyphasic taxon ...200111491349
infection with burkholderia cepacia complex genomovars in patients with cystic fibrosis: virulent transmissible strains of genomovar iii can replace burkholderia multivorans.infection with burkholderia cepacia complex in patients with cystic fibrosis (cf) results in highly variable clinical outcomes. the purpose of this study was to determine if there are genomovar-specific disparities in transmission and disease severity. b. cepacia complex was recovered from 62 patients with cf on > or =1 occasions (genomovar iii, 46 patients; genomovar ii [b. multivorans], 19 patients; genomovar iv [b. stabilis], 1 patient; genomovar v [b. vietnamiensis], 1 patient; and an unclas ...200111588691
pcr-based detection and identification of burkholderia cepacia complex pathogens in sputum from cystic fibrosis patients.pcr amplification of the reca gene followed by restriction fragment length polymorphism (rflp) analysis was investigated for the rapid detection and identification of burkholderia cepacia complex genomovars directly from sputum. successful amplification of the b. cepacia complex reca gene from cystic fibrosis (cf) patient sputum samples containing b. cepacia genomovar i, burkholderia multivorans, b. cepacia genomovar iii, burkholderia stabilis, and burkholderia vietnamiensis was demonstrated. in ...200111724828
characterization of n-acyl homoserine lactone overproducing mutants of burkholderia multivorans atcc 17616.burkholderia multivorans atcc 17616 ordinarily produces insufficient amounts of n-acyl homoserine lactones (ahls) to promote ahl-dependent formation of the pigment violacein by the reporter strain chromobacterium violaceum cv026. we have isolated ahl-overproducing mutants of strain 17616 by screening for variants which do cross-feed ahls to strain cv026. nucleotide-sequence analysis of the bmuir locus which specifies ahl synthase (bmui) and ahl-binding transcriptional activator protein (bmur) in ...200211814664
improved molecular detection of burkholderia cepacia genomovar iii and burkholderia multivorans directly from sputum of patients with cystic fibrosis.optimum detection of the burkholderia cepacia complex (bcc) from sputum of patients with cystic fibrosis (cf) is essential in preventing patient-to-patient transmission of this organism. the aim of this study was to develop an improved pcr assay with reference to sensitivity for the direct detection of bcc organisms from cf sputum employing the reca locus. the sensitivity results of three reca pcr assays were compared using various combinations of previously published primers. these included (i) ...200211830304
evaluation of restriction fragment length polymorphism analysis of 16s rdna as a tool for genomovar characterisation within the burkholderia cepacia complex.a total of 154 burkholderia cepacia complex strains, isolated from cystic fibrosis and non-cystic fibrosis patients and the environment, representing all nine genomovars and a putative tenth, were analysed by 16s rdna-restriction fragment length polymorphism using the restriction enzymes alui, cfoi and ddei. examining this diverse strain collection resulted in very diverse restriction patterns. only b. cepacia genomovar vi could be identified unambiguously. the same restriction patterns were obs ...200212204364
suppression-subtractive hybridisation reveals variations in gene distribution amongst the burkholderia cepacia complex, including the presence in some strains of a genomic island containing putative polysaccharide production genes.some strains of the burkholderia cepacia complex, including the et12 lineage, have been implicated in epidemic spread amongst cystic fibrosis (cf) patients. suppression-subtractive hybridisation was used to identify genomic regions within strain j2315 (et12 lineage; genomovar iiia) that were absent from a non-transmissible genomovar iiib strain. sequence data from 15 subtracted clones were used to interrogate the genome sequence of strain j2315 and identify genomic regions incorporating the subt ...200312610727
lack of correlation between o-serotype, bacteriophage susceptibility and genomovar status in the burkholderia cepacia complex.the burkholderia cepacia complex comprises at least nine phylogenetically related genomic species (genomovars) which cause life-threatening infection in immunocompromised humans, particularly individuals with cystic fibrosis or chronic granulomatous disease. prior to recognition that 'b. cepacia' comprise multiple species, in vitro studies revealed that the lipopolysaccharide (lps) of these gram-negative bacteria is strongly endotoxic. in this study, we used 117 b. cepacia complex isolates to de ...200312628542
fatal outcome of lung transplantation in cystic fibrosis patients due to small-colony variants of the burkholderia cepacia complex.the aim of this study was to investigate the possible role of small-colony variant morphotypes of burkholderia cepacia-like organisms in infectious complications in cystic fibrosis patients following lung transplantation. respiratory tract specimens from 470 cystic fibrosis patients were screened over a 22-month period for burkholderia cepacia-like organisms. nineteen patients were positive for these organisms, eight of whom harboured small-colony-variant morphotypes. three patients underwent bi ...200312687415
distribution and organization of auxotrophic genes on the multichromosomal genome of burkholderia multivorans atcc 17616.the burkholderia multivorans strain atcc 17616 carries three circular chromosomes with sizes of 3.4, 2.5, and 0.9 mb. to determine the distribution and organization of the amino acid biosynthetic genes on the genome of this beta-proteobacterium, various auxotrophic mutations were isolated using a tn5 derivative that was convenient not only for the determination of its insertion site on the genome map but also for the structural analysis of the flanking regions. analysis by pulsed-field gel elect ...200312754231
dna-dna hybridization study of burkholderia species using genomic dna macro-array analysis coupled to reverse genome probing.the present study was aimed at simplifying procedures to delineate species and identify isolates based on dna-dna reassociation. dna macro-arrays harbouring genomic dna of reference strains of several burkholderia species were produced. labelled genomic dna, hybridized to such an array, allowed multiple relative pairwise comparisons. based on the relative dna-dna relatedness values, a complete data matrix was constructed and the ability of the method to discriminate strains belonging to differen ...200312807195
genomovar diversity amongst burkholderia cepacia complex isolates from an australian adult cystic fibrosis unit.in this study, a combination of reca-based pcr assays and 16s rdna restriction fragment length polymorphism (rflp) analysis was used to determine the genomovar diversity of clinical burkholderia cepacia complex isolates. twenty-eight isolates were prospectively collected from patients attending a large australian adult cystic fibrosis (cf) unit, 22 isolates were referred from other australian cf units and a further eight isolates originated from patients without cf. the 28 prospectively collecte ...200312884074
a new mini-transposon for in vivo protein epitope tagging: application to burkholderia multivorans.a short amino acid sequence coding for the mature pseudomonas aeruginosa opri lipoprotein was fused to a mini-tn5 plasposon (mini-transposon with an origin of replication) with tetracycline resistance in order to generate in-frame fusion proteins after transposition. after conjugative transfer to burkholderia multivorans, clones reacting with an anti-opri mab were selected. in-frame opri-tagged proteins were detected and identified for six clones. the six c-tagged proteins were detected by immun ...200312892852
comparative analysis of plant and animal models for characterization of burkholderia cepacia virulence.a simple alfalfa model was developed as an alternative infection model for virulence studies of the burkholderia cepacia complex. symptoms of disease were observed in wounded alfalfa seedlings within 7 days following inoculation of 10(1) to 10(5) cfu of most strains of the b. cepacia complex. strains from seven genomovars of the b. cepacia complex were tested for virulence in the alfalfa model, and the degree of virulence was generally similar in strains belonging to the same genomovar. strains ...200312933878
role of actin filament network in burkholderia multivorans invasion in well-differentiated human airway epithelia.the role of the actin-based cytoskeleton in the internalization process of burkholderia multivorans by well-differentiated human airway epithelia was investigated by immunohistology and confocal microscopy. our data suggest that an intact actin cytoskeleton is required for biofilm formation but not single cell entry or paracytosis.200314573682
in vitro activity and synergy of bismuth thiols and tobramycin against burkholderia cepacia complex.to determine the susceptibility of burkholderia multivorans and burkholderia cenocepacia to bismuth-thiols (bts), and to examine the synergistic effects of tobramycin and subinhibitory concentrations of bts against these organisms.200314585856
evidence of transmission of burkholderia cepacia, burkholderia multivorans and burkholderia dolosa among persons with cystic fibrosis.previous studies have identified specific burkholderia cepacia complex strains that are common to multiple persons with cystic fibrosis (cf). such so-called epidemic strains have an apparent enhanced capacity for inter-patient spread and reside primarily in burkholderia cenocepacia (formerly b. cepacia complex genomovar iii). we sought to identify strains from b. cepacia complex species other than b. cenocepacia that are similarly shared by multiple cf patients. we performed genotype analysis of ...200314612237
improved flagellin genotyping in the burkholderia cepacia complex.oligonucleotide primers designed to n- and c-terminal sequences of burkholderia cepacia complex flic genes were used to amplify and sequence flic genes from a strain of burkholderia vietnamiensis and three isolates of burkholderia multivorans with large flic genes. alignments incorporating the new sequences enabled the design of polymerase chain reaction (pcr) primers for extension of a published pcr/restriction fragment length polymorphism typing method, to include isolates that previously fail ...200314659536
molecular comparison of isolates of burkholderia multivorans from patients with cystic fibrosis in the united kingdom.burkholderia multivorans strains from 47 cystic fibrosis (cf) patients in 28 hospitals were compared by pulsed-field gel electrophoresis (pfge) and flagellin (flic) pcr-restriction fragment length polymorphism (pcr-rflp) analysis. a considerable degree of genetic variation was evident, with each patient harboring a strain with a unique pfge profile. four sizes of flic amplicons were produced, and these amplicons gave 13 rflp types with restriction enzyme mspi. b. multivorans did not appear to sp ...200314662975
conservation of the opcl gene encoding the peptidoglycan-associated outer-membrane lipoprotein among representatives of the burkholderia cepacia complex.members of the burkholderia cepacia complex are gram-negative beta-proteobacteria that are classified into nine genomic species or genomovars. some representatives of this group of bacteria, such as burkholderia multivorans (genomovar ii) and burkholderia cenocepacia (genomovar iii), are considered to be dangerous pathogens for cystic fibrosis (cf) patients because of their capacity to colonize cf lungs. the opcl gene, which encodes the peptidoglycan-associated outer-membrane lipoprotein (pal), ...200415096547
antibacterial effects of knotwood extractives on paper mill bacteria.hydrophilic knotwood extracts from 18 wood species were assessed in disc diffusion and liquid culture tests for antibacterial effects against three species of paper mill bacteria. the pinus sylvestris, p. resinosa, p. contorta, and p. banksiana extracts decreased or inhibited bacterial growth. the susceptibility order was p. sylvestris > p. resinosa > p. contorta > p. banksiana, correlating with the concentrations of pinosylvin and pinosylvin monomethyl ether in these wood species. also, pseudot ...200415112061
proposal to accommodate burkholderia cepacia genomovar vi as burkholderia dolosa sp. nov.phenotypic and genotypic studies revealed new tools for differentiating burkholderia cepacia genomovar vi from burkholderia multivorans and other b. cepacia-complex species. hence, the name burkholderia dolosa sp. nov. is proposed, with lmg 18943(t) (=ccug 47727(t)) as the type strain. b. dolosa can be differentiated from other b. cepacia-complex bacteria by its inability to assimilate tryptamine, azelaic acid and salicin and by its failure to grow on the b. cepacia-selective medium pcat. both 1 ...200415143009
epidemiology of burkholderia cepacia complex species recovered from cystic fibrosis patients: issues related to patient segregation.studies of the prevalence of burkholderia cepacia complex species amongst cystic fibrosis (cf) patients in different geographical regions, and the association between cross-infection and putative transmissibility markers, will further our understanding of these organisms and help to address infection-control issues. in this study, b. cepacia complex isolates from cf patients in different regions of europe were analysed. isolates were examined for b. cepacia complex species and putative transmiss ...200415184539
epidemiology of burkholderia cepacia complex colonisation in cystic fibrosis patients.in belgian cystic fibrosis (cf) clinics, sputum samples are evaluated on selective mast medium routinely every 3 months. in this study, in 1993 and 1999, isolates were further examined by reca restriction fragment length polymorphism analysis and pulsed-field gel electrophoresis of genomic dna restricted with spei. in 1993, 12 patients were colonised with burkholderia cepacia complex (bcc): b. cenocepacia (n=6), b. multivorans (n=3), b. stabilis (n=3). four patients were colonised with the same ...200415218997
two novel clinical presentations of burkholderia cepacia infection.we report two cases of multidrug-resistant burkholderia cepacia (b. cepacia genomovar i) and burkholderia multivorans causing multiple liver abscesses in a patient with bronchial asthma (case 1) and peritonitis in a patient with cirrhosis and hepatitis c virus disease (case 2), respectively. both patients were treated successfully.200415297563
persistence of burkholderia multivorans within the pulmonary macrophage in the murine lung.differences in infection kinetics and host response between burkholderia multivorans and burkholderia cenocepacia were demonstrated in a pulmonary infection model in balb/c mice. b. multivorans persisted in the lung, while b. cenocepacia was cleared. indirect immunofluorescence and electron microscopy of b. multivorans-infected lungs localized bacteria to macrophages. clearance of b. cenocepacia was associated with greater interleukin-1beta and neutrophil responses than the responses induced by ...200415385520
clinical outcome of burkholderia cepacia complex infection in cystic fibrosis adults.the burkholderia cepacia complex (bcc) is one of the most important groups of organisms infecting cystic fibrosis (cf) patients. the aim of the study was to examine how infection with bcc affects clinical outcome.200415463892
'cepacia syndrome' with burkholderia multivorans, 9 years after initial colonization.a 16-year-old boy with cystic fibrosis developed 'cepacia syndrome' 9 years after the first isolation of burkholderia multivorans. it is important to recognise that 'cepacia syndrome' is not restricted to those infected with genomovar type iii strains and that rapid, irreversible clinical decline can occur many years after the 1st isolation of burkholderia cepacia complex (bcc).200415463897
species distribution and ribotype diversity of burkholderia cepacia complex isolates from french patients with cystic fibrosis.a total of 153 burkholderia cepacia strains obtained from 153 french patients with cystic fibrosis were identified as burkholderia multivorans (51.6%) or burkholderia cenocepacia (45.1%). eighty-two genotypes were identified using pvuii and ecori ribotyping. b. multivorans genotype a (found in 32 french patients) and two other genotypes were also identified among isolates from austrian, german, italian, and canadian patients.200415472352
burkholderia cenocepacia and burkholderia multivorans: influence on survival in cystic fibrosis.burkholderia cepacia infection has been associated with a poor prognosis for patients with cystic fibrosis (cf). it is now recognised that organisms classified as b cepacia comprise a number of distinct genomic species each known as a genomovar of the b cepacia complex (bcc). the outcome of infection for cf patients with individual genomovars is unknown. the clinical outcome of infection with the two most commonly isolated genomovars (b cenocepacia and b multivorans) was studied at a specialist ...200415516469
long-term outcome of lung transplantation for cystic fibrosis--danish results.over the last decades improvements in medical therapies have delayed the progression of lung disease in cystic fibrosis (cf). however, lung disease is still the most common cause of premature death, and lung transplantation today is the only treatment for end-stage lung disease in patients with cf. we present a retrospective review of the outcome of cf patients transplanted in denmark since start of the national lung transplantation programme in 1992.200415541981
chronic burkholderia multivorans bronchial infection in a non-cystic fibrosis individual with mannose binding lectin deficiency.the case history is presented of a woman with multiple respiratory infections and mannose binding lectin (mbl) deficiency but no evidence of bronchiectasis who developed a chronic burkholderia multivorans infection. careful microbiological assessment is needed in patients with recurrent respiratory infection and the presence of b multivorans should trigger further immunological investigation including assessment of mbl status.200515681508
single-step purification of lipase from burkholderia multivorans using polypropylene matrix.lipase from burkholderia multivorans was purified with high yields directly from fermentation broth by a single-step purification protocol involving adsorption and desorption. the crude enzyme (lyophilized powder) from b. multivorans was loaded on accurel (membrana, germany), a polypropylene matrix, using butanol as the solvent in a buffer at ph 9.0 and ambient temperature for a period of 12 h. the enzyme adsorbed onto the matrix with high specific activity (33 units mg(-1) protein). this was fo ...200515711795
high-temperature-induced transposition of insertion elements in burkholderia multivorans atcc 17616.an efficient and quantitative method to analyze the transposition of various insertion sequence (is) elements in burkholderia multivorans atcc 17616 was devised. pgen500, a plasmid carrying a bacillus subtilis-derived sacb gene, was introduced into atcc 17616 cells, and 25% of their sucrose-resistant derivatives were found to carry various is elements on pgen500. a pcr-based experimental protocol, in which a mixture of several specific primer pairs was used, revealed that pgen500 captured, in ad ...200515812007
use of suppression-subtractive hybridization to identify genes in the burkholderia cepacia complex that are unique to burkholderia cenocepacia.we have previously shown differences in virulence between species of the burkholderia cepacia complex using the alfalfa infection model and the rat agar bead chronic infection model. burkholderia cenocepacia strains were more virulent in these two infection models than burkholderia multivorans and burkholderia stabilis strains. in order to identify genes that may account for the increased virulence of b. cenocepacia, suppression-subtractive hybridization was performed between b. cenocepacia k56- ...200516030222
the nematode panagrellus redivivus is susceptible to killing by human pathogens at 37 degrees c.caenorhabditis elegans has been used as a host for the study of bacteria that cause disease in mammals. however, a significant limitation of the model is that c. elegans is not viable at 37 degrees c. we report that the gonochoristic nematode panagrellus redivivus survives at 37 degrees c and maintains its life cycle at temperatures up to and including 31.5 degrees c. the c. elegans pathogens pseudomonas aeruginosa, salmonella enterica, staphylococcus aureus, but not yersinia pseudotuberculosis, ...200516040202
[use of 16s rrna gene sequencing for identification of "pseudomonas-like" isolates from sputum of patients with cystic fibrosis].since nonfermenting, gram negative bacilli recovered from patients with cystic fibrosis could be misidentified with phenotypic procedures, we used partial 16s ribosomal rna gene (16s gene) sequencing to identify these "pseudomonas-like" isolates. 473 isolates were recovered from 66 patients in 2003. sequencing was used to identify 29 (from 24 patients) of the 473 isolates, showing unclear results with routine tests. pcr with specific primers was carried out to amplify a 995 bp fragment, which wa ...200516081224
update on the burkholderia cepacia complex.the burkholderia cepacia complex is comprised of a group of related bacterial species that are capable of causing life-threatening respiratory tract infection in persons with cystic fibrosis. this article reviews advances in our understanding of burkholderia cepacia complex infection in cystic fibrosis, focusing on the taxonomy, clinical microbiology, and epidemiology, as well as the natural history and clinical outcomes associated with burkholderia cepacia complex infection.200516217180
transient isolation of burkholderia multivorans and burkholderia cenocepacia from a brazilian cystic fibrosis patient chronically colonized with burkholderia vietnamiensis.fifteen serial burkholderia cepacia complex isolates recovered over a period of 4 years from a single cystic fibrosis patient were analysed for genomovar status by means of reca sequence determination, and genetic relatedness by rapd-pcr. twelve isolates were assigned as burkholderia vietnamiensis, two as burkholderia cenocepacia and one as burkholderia multivorans. b. vietnamiensis persisted in the airways during 4 years, except in three occasions when b. cenocepacia or b. multivorans were isol ...200516266831
distribution and genomic location of active insertion sequences in the burkholderia cepacia complex.this study aimed firstly to establish the distribution and copy number within the burkholderia cepacia complex of three insertion sequences (is402, is407 and is1416) that possess the ability to activate transcription and hence influence gene expression. a second aim was to map the genomic insertion sites of one of the active insertion sequences (is407) to establish putative links between insertion site and downstream gene activation. the resulting data revealed that all three insertion sequences ...200616388024
identification of burkholderia cepacia complex pathogens by rapid-cycle pcr with fluorescent hybridization probes.members of the burkholderia cepacia complex are important bacterial pathogens in cystic fibrosis (cf) patients. the b. cepacia complex currently consists of nine genetic subgroups (genomovars) of different epidemiological relevance and possibly of different pathogenic potential in humans. in this study, a new approach was developed for the rapid identification of b. cepacia genomovar i, burkholderia multivorans (genomovar ii), burkholderia cenocepacia (lineage iii-a and iii-b), burkholderia stab ...200616687590
invasion and biofilm formation of burkholderia dolosa is comparable with burkholderia cenocepacia and burkholderia multivorans.colonisation with burkholderia cepacia complex pathogens has been associated with accelerated decline in cystic fibrosis (cf) patients. the two most common species among the cf community are burkholderia cenocepacia and burkholderia multivorans. however, burkholderia dolosa has recently been causing concern due to its transmissibility and virulence in cf patients.200716781896
exopolysaccharides produced by clinical strains belonging to the burkholderia cepacia complex.in the frame of a research line dedicated to better clarify the role of exopolysaccharides (eps) in bacterial virulence, eps produced by species of the burkholderia cepacia complex (bcc), namely burkholderia multivorans, burkholderia cenocepacia, and a bcc member of undetermined genomovar, all isolated at the cystic fibrosis regional centre of florence (italy), were investigated for they structural properties.200716860003
highly purified lipopolysaccharides from burkholderia cepacia complex clinical isolates induce inflammatory cytokine responses via tlr4-mediated mapk signalling pathways and activation of nfkappab.in cystic fibrosis (cf), bacteria of the burkholderia cepacia complex (bcc) can induce a fulminant inflammation with pneumonitis and sepsis. lipopolysaccharide (lps) may be an important virulence factor associated with this decline but little is known about the molecular pathogenesis of bcc lps. in this study we have investigated the inflammatory response to highly purified lps from different bcc clinical isolates and the cellular signalling pathways employed. the inflammatory response (tnfalpha ...200717002785
evidence of coinfection with distinct strains of burkholderia multivorans in a cystic fibrosis patient.we report a cystic fibrosis patient with chronic burkholderia multivorans infection involving persistency of one strain and temporary, consecutive coinfection with two different strains. comparison of the colony morphology with the genotype revealed no correlation. these data are important for interpretation of clinical outcome and transmission studies in cf patients.200617033756
[isolation and identification of burkholderia cepacia complex from cystic fibrosis patients].in this study, sputum samples collected from cystic fibrosis patients with preliminary diagnosis of pulmonary infection were evaluated for the presence of bacteria belonging to burkholderia cepacia complex by commercial phenotypic systems and reca polymerase chain reaction (pcr). a total of 85 patients ages between 5-30 years (mean age: 12.8 years) were included to the study with female/male ratio of 44/41. the mostly isolated bacteria from 85 sputum samples were found as staphylococcus aureus ( ...200617205687
nasal immunization with burkholderia multivorans outer membrane proteins and the mucosal adjuvant adamantylamide dipeptide confers efficient protection against experimental lung infections with b. multivorans and b. cenocepacia.chronic lung infection by opportunistic pathogens, such as pseudomonas aeruginosa and members of the burkholderia cepacia complex, is a major cause of morbidity and mortality in patients with cystic fibrosis. outer membrane proteins (omps) of gram-negative bacteria are promising vaccine antigen candidates. in this study, we evaluated the immunogenicity, protection, and cross-protection conferred by intranasal vaccination of mice with omps from b. multivorans plus the mucosal adjuvant adamantylam ...200717296759
pyrosequencing of a reca gene variable region for burkholderia cepacia complex genomovar identification.we developed an assay to identify burkholderia cepacia complex genomovars by pyrosequencing of a variable reca gene segment. fifteen reference strains and 30 clinical isolates of b. cepacia complex were sequenced. full 77-base pair target sequences were obtained from 44 of the 45 isolates, and blast queries of the sequences correctly identified the genomovar of these 44 isolates. three burkholderia multivorans isolates were identified as b. multivorans/burkholderia ambifaria, indicating that add ...200717509790
the effect of recombinant human lactoferrin on growth and the antibiotic susceptibility of the cystic fibrosis pathogen burkholderia cepacia complex when cultured planktonically or as biofilms.the cystic fibrosis (cf) pathogen burkholderia cepacia complex (bcc) is innately resistant to antibiotics and the development of effective therapeutic strategies to treat these infections is a major challenge. the objectives of this study were to investigate the effects of recombinant human lactoferrin (rhl) on planktonic and biofilm cultures of bcc organisms and to establish whether lactoferrin alters the susceptibility of these cultures to a range of antibiotic therapies.200717595284
a german external quality survey of diagnostic microbiology of respiratory tract infections in patients with cystic fibrosis.the goal of this pilot study was to design an external quality assessment (eqa) scheme for german cystic fibrosis (cf) clinical microbiology laboratories. therefore, a multicentre study of 18 german cf laboratories was performed to evaluate their proficiency in analyzing cf respiratory secretions.200817658302
environmental persistence of pseudomonas aeruginosa and burkholderia multivorans in sea water: preliminary evidence of a viable but non-culturable state. 200717910284
a novel recombinant ethyl ferulate esterase from burkholderia multivorans.isolation and identification of bacterial isolates with specific ferulic acid (fa) esterase activity and cloning of a gene encoding activity.200717953572
elucidating global epidemiology of burkholderia multivorans in cases of cystic fibrosis by multilocus sequence typing.burkholderia multivorans is a prominent b. cepacia complex (bcc) species causing infection in people with cystic fibrosis. despite infection control measures being introduced to reduce the spread of bcc there is a continued emergence of infections by b. multivorans. our objective was to analyze a global collection of b. multivorans isolates, comparing those from environmental and clinical sources with those from reported outbreaks. multilocus sequence typing (mlst) was performed on 107 b. multiv ...200818032622
cepacia-like syndrome caused by burkholderia multivorans.the variable severity of burkholderia cepacia complex infections in cystic fibrosis (cf) has recently been ascribed to differences in the virulence between genomovars. specifically, genomovar iii isolates have been associated with higher transmission rates and adverse outcomes compared to other b cepacia genomovars, and consequently further segregation between genomovar iii and non-genomovar iii b cepacia infected patients is advocated in some centres. the important role of non-genomovar iii iso ...200318159435
burkholderia cepacia complex bacteria: opportunistic pathogens with important natural biology.interaction with plants around their roots and foliage forms the natural habitat for a wide range of gram-negative bacteria such as burkholderia, pseudomonas and ralstonia. during these interactions many of these bacteria facilitate highly beneficial processes such as the breakdown of pollutants or enhancement of crop growth. all these bacterial species are also capable of causing opportunistic infections in vulnerable individuals, especially people with cystic fibrosis (cf). here we will review ...200818217926
resistance of planktonic and biofilm-grown burkholderia cepacia complex isolates to the transition metal gallium.the burkholderia cepacia complex is a group of pathogens that can cause severe pulmonary infections in cystic fibrosis (cf) patients. the aim of the present study was to investigate the in vitro activity of gallium against planktonic and biofilm-grown b. cepacia complex isolates.200818310047
differential modulation of innate immune cell functions by the burkholderia cepacia complex: burkholderia cenocepacia but not burkholderia multivorans disrupts maturation and induces necrosis in human dendritic cells.burkholderia cepacia complex (bcc) bacteria cause pulmonary infections that can evolve into fatal overwhelming septicemia in chronic granulomatous disease or cystic fibrosis patients. burkholderia cenocepacia and burkholderia multivorans are responsible for the majority of bcc infections in cystic fibrosis patients, but b. cenocepacia is generally associated with a poorer prognosis than b. multivorans. the present study investigated whether these pathogens could modulate the normal functions of ...200818624797
the structure and proinflammatory activity of the lipopolysaccharide from burkholderia multivorans and the differences between clonal strains colonizing pre and posttransplanted lungs.the burkholderia cepacia complex is a group of gram-negative bacteria that are opportunistic pathogens for humans especially in cystic fibrosis patients. lipopolysaccharide (lps) molecules are potent virulence factors of gram-negative bacteria organisms essential for bacterial survival. a complete analysis of the bacterial lipopolysaccharide structure to function relationship is required to understand the chemical basis of the inflammatory process. we have therefore investigated the structures o ...200818684854
use of quorum sensing inhibitors to interfere with biofilm formation and development in burkholderia multivorans and burkholderia cenocepacia.burkholderia cepacia complex strains are opportunistic pathogens causing life-threatening infections in cystic fibrosis patients. b. cepacia complex strains are resistant to many antimicrobial agents and commonly produce biofilms in vitro and in vivo. this contributes to their virulence and makes burkholderia infections difficult to treat. recently, the quorum sensing (qs) system of burkholderia spp. has been found to affect their biofilm-forming ability, making it an attractive target for antim ...200919146953
biocide susceptibility of the burkholderia cepacia complex.the burkholderia cepacia complex (bcc) species are important opportunistic pathogens with intrinsic antibiotic resistance. they are also well known as contaminants of disinfectants, yet their biocide susceptibility has not been studied in detail. we investigated bcc biocide susceptibility and correlated it to their taxonomy, antibiotic susceptibility and ability to form biofilms.200919153076
use of fourier transform infrared spectroscopy and chemometrics to discriminate clinical isolates of bacteria of the burkholderia cepacia complex from different species and ribopatterns.a methodology for the discrimination of burkholderia cepacia complex (bcc) clinical isolates at the species level and at the ribopattern level using fourier transform infrared (ftir) spectroscopy and chemometrics analysis was assessed in this study. different bcc sequential isolates collected at the santa maria hospital (hsm), in portugal, from clinically infected cystic fibrosis (cf) patients were previously classified by established molecular methods at the species level and differentiated at ...200919579020
virulence and cellular interactions of burkholderia multivorans in chronic granulomatous disease.chronic granulomatous disease (cgd) patients are susceptible to life-threatening infections by the burkholderia cepacia complex. we used leukocytes from cgd and healthy donors and compared cell association, invasion, and cytokine induction by burkholderia multivorans strains. a cgd isolate, cgd1, showed higher cell association than that of an environmental isolate, env1, which correlated with cell entry. all b. multivorans strains associated significantly more with cells from cgd patients than w ...200919635825
distribution of cepacian biosynthesis genes among environmental and clinical burkholderia strains and role of cepacian exopolysaccharide in resistance to stress conditions.the genus burkholderia includes strains pathogenic to animals and plants, bioremediators, or plant growth promoters. genome sequence analyses of representative burkholderia cepacia complex (bcc) and non-bcc strains for the presence of the bce-i gene cluster, directing the biosynthesis of the exopolysaccharide (eps) cepacian, further extended this previously described cluster by another 9 genes. the genes in the bce-ii cluster were named bcem to bceu and encode products putatively involved in nuc ...201019948863
approaches to measure the fitness of burkholderia cepacia complex isolates.members of the burkholderia cepacia complex (bcc) are highly resistant to many antibacterial agents and infection can be difficult to eradicate. a coordinated approach has been used to measure the fitness of bcc bacteria isolated from cystic fibrosis (cf) patients with chronic bcc infection using methods relevant to bcc growth and survival conditions. significant differences in growth rate were observed among isolates; slower growth rates were associated with isolates that exhibited higher mics ...201020185551
surface hydrophobicity of petroleum hydrocarbon degrading burkholderia strains and their interactions with napls and surfaces.bacterial cell surface hydrophobicity (csh) is an important factor governing the growth and adhesion behavior of microorganisms on non-aqueous phase liquids (napls). in this work csh and surface charge was quantified for three oil degrading burkholderia cultures: aliphatic degrader burkholderia cepacia (es1) and two strains of aromatic degrading burkholderia multivorans (ng1 and hn1) based on contact angle and zeta potential measurement. model non-aqueous phase liquids (napls) were formulated us ...201020236810
real-time pcr method for the quantification of burkholderia cepacia complex attached to lung epithelial cells and inhibition of that attachment.to develop a rapid method to quantify the attachment of the cystic fibrosis pathogen, burkholderia multivorans, to lung epithelial cells (16hbe14o(-)) using real-time pcr with a view to monitoring potential inhibition of lung cell attachment.201020337933
antisense phosphorodiamidate morpholino oligomers targeted to an essential gene inhibit burkholderia cepacia complex.members of the burkholderia cepacia complex (bcc) cause considerable morbidity and mortality in patients with chronic granulomatous disease and cystic fibrosis. many bcc strains are antibiotic resistant, which requires the exploration of novel antimicrobial approaches, including antisense technologies such as phosphorodiamidate morpholino oligomers (pmos).201020438352
structural and functional characterization of diffusible signal factor family quorum-sensing signals produced by members of the burkholderia cepacia complex.previous work has shown that burkholderia cenocepacia produces the diffusible signal factor (dsf) family signal cis-2-dodecenoic acid (c(12):delta(2), also known as bdsf), which is involved in the regulation of virulence. in this study, we determined whether c(12):delta(2) production is conserved in other members of the burkholderia cepacia complex (bcc) by using a combination of high-performance liquid chromatography, mass spectrometry, and bioassays. our results show that five bcc species are ...201020511428
unusual distribution of burkholderia cepacia complex species in danish cystic fibrosis clinics may stem from restricted transmission between patients.forty-four of 48 burkholderia cepacia complex strains cultured from danish cystic fibrosis patients were burkholderia multivorans, a distribution of species that has not been reported before. although cases of cross infections were demonstrated, no major epidemic clone was found. the species distribution may represent the sporadic acquisition of bacteria from the environment.201020519474
macrophage responses to cf pathogens: jnk map kinase signaling by burkholderia cepacia complex lipopolysaccharide.chronic bacterial colonization of the airways with opportunistic pathogens is the primary cause of morbidity and mortality in cystic fibrosis (cf) patients. burkholderia cepacia complex (bcc) organisms pose a particular challenge in cf lung disease, due in part to their ability to trigger a fulminant pneumonia. this study compares the u937 macrophage response to two bcc species, b. cenocepacia and burkholderia multivorans, against pseudomonas aeruginosa and staphylococcus aureus. the two bcc str ...201020602636
a decade of burkholderia cenocepacia virulence determinant research.the burkholderia cepacia complex (bcc) is a group of genetically related environmental bacteria that can cause chronic opportunistic infections in patients with cystic fibrosis (cf) and other underlying diseases. these infections are difficult to treat due to the inherent resistance of the bacteria to antibiotics. bacteria can spread between cf patients through social contact and sometimes cause cepacia syndrome, a fatal pneumonia accompanied by septicemia. burkholderia cenocepacia has been the ...201020643851
identification of potential diagnostic markers among burkholderia cenocepacia and b. multivorans supernatants.patients with cystic fibrosis (cf) are susceptible to chronic respiratory infections with a number of bacterial pathogens. among them, the burkholderia cepacia complex (bcc) bacteria, consisting of nine related species, have emerged as problematic cf pathogens due to their antibiotic resistance, incidence of nosocomial infection, and person-to-person transmission. bcc organisms present the clinical microbiologist with a diagnostic dilemma due to the lack of phenotypic biochemical or growth-relat ...201020810766
invasion of burkholderia cepacia complex isolates into lung epithelial cells involves glycolipid receptors.burkholderia cepacia complex (bcc) is a group of opportunistic cystic fibrosis (cf) pathogens that invade lung epithelial cells. the mechanisms of invasion are poorly understood, in particular, the receptors utilised by this bacterium in the invasion process have not been identified. the aim of this study was to investigate the epithelial receptors involved in the invasion of bcc isolates. we confirmed that invasion into two lung epithelial cell lines (16hbe14o- and cfbe41o-) which have a non-cf ...201020826205
burkholderia cenocepacia in cystic fibrosis: epidemiology and molecular mechanisms of virulence.burkholderia cepacia complex (bcc) bacteria have gained notoriety as pathogens in cystic fibrosis (cf) because they are difficult to identify and treat, and also have the ability to spread between cf individuals. of the 17 formally named species within the complex, burkholderia multivorans and burkholderia cenocepacia dominate in cf. multilocus sequence typing has proven to be a very useful tool for tracing the global epidemiology of bcc bacteria and has shown that b. cenocepacia strains with hi ...201020880411
functional quorum sensing systems are maintained during chronic burkholderia cepacia complex infections in patients with cystic fibrosis.quorum sensing (qs) contributes to the virulence of pseudomonas aeruginosa and burkholderia cepacia complex lung infections. p. aeruginosa qs mutants are frequently isolated from patients with cystic fibrosis. the objective of this study was to determine whether similar adaptations occur over time in b. cepacia complex isolates. forty-five burkholderia multivorans and burkholderia cenocepacia sequential isolates from patients with cystic fibrosis were analyzed for n-acyl-homoserine lactone activ ...201021208930
diversity analysis of burkholderia cepacia complex in the water bodies of west lake, hangzhou, china.a survey of burkholderia cepacia complex (bcc) species was conducted in water bodies of west lake in china. a total of 670 bacterial isolates were recovered on selective media. out of them, 39.6% (265 isolates) were assigned to the following species: burkholderia multivorans, burkholderia cenocepacia reca lineage iiia, iiib, burkholderia stabilis, burkholderia vietnamiensis, and burkholderia seminalis while b. cenocepacia is documented as a dominant bcc species in water of west lake. in addition ...201121538256
mucoid morphotype variation of burkholderia multivorans during chronic cystic fibrosis lung infection is correlated with changes in metabolism, motility, biofilm formation and virulence.burkholderia cepacia complex (bcc) bacteria are opportunistic pathogens infecting hosts such as cystic fibrosis (cf) patients. bcc long-term infection of cf patient airways has been associated with emergence of phenotypic variation. here we studied two burkholderia multivorans clonal isolates displaying different morphotypes from a chronically infected cf patient to evaluate traits development during lung infection. expression profiling of mucoid d2095 and nonmucoid d2214 isolates revealed decre ...201121835880
resistance of the burkholderia cepacia complex to fosmidomycin and fosmidomycin derivatives.the burkholderia cepacia complex (bcc) is a group of 17 closely related opportunistic pathogens that are able to infect the respiratory tract of cystic fibrosis patients. bcc bacteria are intrinsically resistant to many antibiotics and are therefore difficult to eradicate. fosmidomycin could be a new therapeutic agent to treat bcc infections as it inhibits 1-deoxy-d-xylulose-5-phosphate reductoisomerase (dxr), a key enzyme in the non-mevalonate pathway essential in bcc bacteria for isoprenoid sy ...201121724375
Alteration in cell surface properties of Burkholderia spp. during surfactant-aided biodegradation of petroleum hydrocarbons.Chemical surfactants may impact microbial cell surface properties, i.e., cell surface hydrophobicity (CSH) and cell surface charge, and may thus affect the uptake of components from non-aqueous phase liquids (NAPLs). This work explored the impact of Triton X-100, Igepal CA 630, and Tween 80 (at twice the critical micelle concentration, CMC) on the cell surface characteristics of Burkholderia cultures, Burkholderia cepacia (ES1, aliphatic degrader) and Burkholderia multivorans (NG1, aromatic degr ...201122089390
Screening microorganisms for insulin binding reveals binding by Burkholderia multivorans and Burkholderia cenocepacia and novel attachment of insulin to Aeromonas salmonicida via the A-layer.Exposure to microorganisms is considered an environmental factor which can contribute to Type 1 diabetes. Insulin binding proteins on microorganisms may induce production of antibodies which can react with the human insulin receptor (HIR) with possible consequences in developing a diabetic autoimmune response against HIR and insulin. The interaction of insulin with microorganisms was studied by screening forty-five microbial species for their ability to bind insulin. Binding assays were performe ...201122171975
[pneumonia-causing bacterial pathogens in intensive care patients].this prospective study aimed at determining etiologic agents causing nosocomial pneumonia in a precisely defined group of patients and resistance of bacterial pathogens to antimicrobial drugs.201122052100
characterization of the poly-β-1,6-n-acetylglucosamine polysaccharide component of burkholderia biofilms.we demonstrated the production of poly-β-1,6-n-acetylglucosamine (pnag) polysaccharide in the biofilms of burkholderia multivorans, burkholderia vietnamiensis, burkholderia ambifaria, burkholderia cepacia, and burkholderia cenocepacia using an immunoblot assay for pnag. these results were confirmed by further studies, which showed that the pnag hydrolase, dispersin b, eliminated immunoreactivity of extracts from the species that were tested (b. cenocepacia and b. multivorans). dispersin b also i ...201121984237
Identification of Hopanoid Biosynthesis Genes Involved in Polymyxin Resistance in Burkholderia multivorans.A major challenge to clinical therapy of Burkholderia cepacia complex (Bcc) pulmonary infections is their innate resistance to a broad range of antimicrobials, including polycationic agents such as aminoglycosides, polymyxins, and cationic peptides. To identify genetic loci associated with this phenotype, a transposon mutant library was constructed in B. multivorans ATCC 17616 and screened for increased susceptibility to polymyxin B. Compared to the parent strain, mutant 26D7 exhibited 8- and 16 ...201222006009
cepacia syndrome in a cystic fibrosis patient colonised with burkholderia multivorans. 201122715269
immunoproteomic analysis of proteins expressed by two related pathogens, burkholderia multivorans and burkholderia cenocepacia, during human infection.burkholderia cepacia complex (bcc) is an opportunistic bacterial pathogen that causes chronic infections in people with cystic fibrosis (cf). it is a highly antibiotic resistant organism and bcc infections are rarely cleared from patients, once they are colonized. the two most clinically relevant species within bcc are burkholderia cenocepacia and burkholderia multivorans. the virulence of these pathogens has not been fully elucidated and the virulence proteins expressed during human infection h ...201324260482
comparative metabolic systems analysis of pathogenic burkholderia.burkholderia cenocepacia and burkholderia multivorans are opportunistic drug-resistant pathogens that account for the majority of burkholderia cepacia complex infections in cystic fibrosis patients and also infect other immunocompromised individuals. while they share similar genetic compositions, b. cenocepacia and b. multivorans exhibit important differences in pathogenesis. we have developed reconciled genome-scale metabolic network reconstructions of b. cenocepacia j2315 and b. multivorans at ...201424163337
can early burkholderia cepacia complex infection in cystic fibrosis be eradicated with antibiotic therapy?organisms of the burkholderia cepacia complex (bcc) are important pathogens in cystic fibrosis (cf). the majority of those who acquire bcc develop chronic infection but it can also result in rapid decline in a significant minority. in addition, chronic infection with burkholderia cenocepacia in particular is regarded as a contraindication to lung transplantation in many units. whilst aggressive antibiotic therapy is employed in cf to eradicate pseudomonas aeruginosa before infection becomes irre ...201122919584
draft genome sequence determination for cystic fibrosis and chronic granulomatous disease burkholderia multivorans isolates.burkholderia multivorans is a gram-negative bacterium and a member of the burkholderia cepacia complex, which is frequently associated with respiratory infections in people with cystic fibrosis (cf) and chronic granulomatous disease (cgd). we are reporting the genome sequences of 4 b. multivorans strains, 2 from cf patients and 2 from cgd patients.201223105085
stress conditions triggering mucoid morphotype variation in burkholderia species and effect on virulence in galleria mellonella and biofilm formation in vitro.burkholderia cepacia complex (bcc) bacteria are opportunistic pathogens causing chronic respiratory infections particularly among cystic fibrosis patients. during these chronic infections, mucoid-to-nonmucoid morphotype variation occurs, with the two morphotypes exhibiting different phenotypic properties. here we show that in vitro, the mucoid clinical isolate burkholderia multivorans d2095 gives rise to stable nonmucoid variants in response to prolonged stationary phase, presence of antibiotics ...201324358195
genomic characterization of jg068, a novel virulent podovirus active against burkholderia cenocepacia.as is true for many other antibiotic-resistant gram-negative pathogens, members of the burkholderia cepacia complex (bcc) are currently being assessed for their susceptibility to phage therapy as an antimicrobial treatment. the objective of this study was to perform genomic and limited functional characterization of the novel bcc phage jg068 (vb_bcep_jg068).201323978260
development of a multiple-locus variable-number tandem-repeat typing scheme for genetic fingerprinting of burkholderia cenocepacia and application to nationwide epidemiological analysis.organisms of the burkholderia cepacia complex are especially important pathogens in cystic fibrosis (cf), with a propensity for patient-to-patient spread and long-term respiratory colonization. b. cenocepacia and burkholderia multivorans account for the majority of infections in cf, and major epidemic clones have been recognized throughout the world. the aim of the present study was to develop and evaluate a multilocus variable-number tandem-repeat (vntr) analysis (mlva) scheme for b. cenocepaci ...201425411181
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