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1.
J Med Virol ; 95(1): e28423, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36546412

RESUMO

The SARS-CoV-2 variant Omicron emerged in late 2021. In British Columbia (BC), Canada, and globally, three genetically distinct subvariants of Omicron, BA.1, BA.2, and BA.5, emerged and became dominant successively within an 8-month period. SARS-CoV-2 subvariants continue to circulate in the population, acquiring new mutations that have the potential to alter infectivity, immunity, and disease severity. Here, we report a propensity-matched severity analysis from residents of BC over the course of the Omicron wave, including 39,237 individuals infected with BA.1, BA.2, or BA.5 based on paired high-quality sequence data and linked to comprehensive clinical outcomes data between December 23, 2021 and August 31, 2022. Relative to BA.1, BA.2 cases were associated with a 15% and 28% lower risk of hospitalization and intensive care unit (ICU) admission (aHRhospital = 1.17; 95% confidence interval [CI] = 1.096-1.252; aHRICU = 1.368; 95% CI = 1.152-1.624), whereas BA.5 infections were associated with an 18% higher risk of hospitalization (aHRhospital = 1.18; 95% CI = 1.133-1.224) after accounting for age, sex, comorbidities, vaccination status, geography, and social determinants of health. Phylogenetic analysis revealed no specific subclades associated with more severe clinical outcomes for any Omicron subvariant. In summary, BA.1, BA.2, and BA.5 subvariants were associated with differences in clinical severity, emphasizing how variant-specific monitoring programs remain critical components of patient and population-level public health responses as the pandemic continues.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Colúmbia Britânica/epidemiologia , SARS-CoV-2/genética , Estudos de Coortes , Filogenia , COVID-19/epidemiologia
2.
BMC Genomics ; 23(1): 710, 2022 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-36258173

RESUMO

BACKGROUND: The COVID-19 pandemic remains a global public health concern. Advances in sequencing technologies has allowed for high numbers of SARS-CoV-2 whole genome sequence (WGS) data and rapid sharing of sequences through global repositories to enable almost real-time genomic analysis of the pathogen. WGS data has been used previously to group genetically similar viral pathogens to reveal evidence of transmission, including methods that identify distinct clusters on a phylogenetic tree. Identifying clusters of linked cases can aid in the regional surveillance and management of the disease. In this study, we present a novel method for producing stable genomic clusters of SARS-CoV-2 cases, cov2clusters, and compare the accuracy and stability of our approach to previous methods used for phylogenetic clustering using real-world SARS-CoV-2 sequence data obtained from British Columbia, Canada. RESULTS: We found that cov2clusters produced more stable clusters than previously used phylogenetic clustering methods when adding sequence data through time, mimicking an increase in sequence data through the pandemic. Our method also showed high accuracy when predicting epidemiologically informed clusters from sequence data. CONCLUSIONS: Our new approach allows for the identification of stable clusters of SARS-CoV-2 from WGS data. Producing high-resolution SARS-CoV-2 clusters from sequence data alone can a challenge and, where possible, both genomic and epidemiological data should be used in combination.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Pandemias , COVID-19/epidemiologia , Filogenia , Genoma Viral , Genômica , Análise por Conglomerados
3.
Genome Res ; 27(4): 650-662, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28325850

RESUMO

Chronic bacterial infections of the lung are the leading cause of morbidity and mortality in cystic fibrosis patients. Tracking bacterial evolution during chronic infections can provide insights into how host selection pressures-including immune responses and therapeutic interventions-shape bacterial genomes. We carried out genomic and phenotypic analyses of 215 serially collected Burkholderia cenocepacia isolates from 16 cystic fibrosis patients, spanning a period of 2-20 yr and a broad range of epidemic lineages. Systematic phenotypic tests identified longitudinal bacterial series that manifested progressive changes in liquid media growth, motility, biofilm formation, and acute insect virulence, but not in mucoidy. The results suggest that distinct lineages follow distinct evolutionary trajectories during lung infection. Pan-genome analysis identified 10,110 homologous gene clusters present only in a subset of strains, including genes restricted to different molecular types. Our phylogenetic analysis based on 2148 orthologous gene clusters from all isolates is consistent with patient-specific clades. This suggests that initial colonization of patients was likely by individual strains, followed by subsequent diversification. Evidence of clonal lineages shared by some patients was observed, suggesting inter-patient transmission. We observed recurrent gene losses in multiple independent longitudinal series, including complete loss of Chromosome III and deletions on other chromosomes. Recurrently observed loss-of-function mutations were associated with decreases in motility and biofilm formation. Together, our study provides the first comprehensive genome-phenome analyses of B. cenocepacia infection in cystic fibrosis lungs and serves as a valuable resource for understanding the genomic and phenotypic underpinnings of bacterial evolution.


Assuntos
Infecções por Burkholderia/microbiologia , Burkholderia cenocepacia/genética , Fibrose Cística/microbiologia , Fenótipo , Polimorfismo Genético , Adolescente , Animais , Biofilmes , Infecções por Burkholderia/complicações , Burkholderia cenocepacia/isolamento & purificação , Burkholderia cenocepacia/patogenicidade , Burkholderia cenocepacia/fisiologia , Criança , Pré-Escolar , Fibrose Cística/complicações , Genótipo , Humanos , Pulmão/microbiologia , Mariposas/microbiologia , Virulência , Adulto Jovem
4.
Am J Transplant ; 19(3): 933-938, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30091842

RESUMO

"Cepacia syndrome", caused by Burkholderia cepacia complex and often associated with cystic fibrosis, carries a high mortality rate. It is rare for Burkholderia multivorans, a species within the B. cepacia complex, to cause cepacia syndrome even among patients with cystic fibrosis. This is the first reported fatal case of cepacia syndrome caused by B. multivorans occurring in a pediatric liver transplant recipient who does not have cystic fibrosis. We describe the unique characteristics of this pathogen among the non-cystic fibrosis population and the importance of early recognition and treatment.


Assuntos
Infecções por Burkholderia/microbiologia , Complexo Burkholderia cepacia/patogenicidade , Febre de Causa Desconhecida/cirurgia , Transplante de Fígado/efeitos adversos , Sepse/etiologia , Infecções por Burkholderia/complicações , Evolução Fatal , Febre de Causa Desconhecida/patologia , Humanos , Lactente , Masculino , Sepse/patologia
5.
Antimicrob Agents Chemother ; 60(1): 348-55, 2016 01.
Artigo em Inglês | MEDLINE | ID: mdl-26503664

RESUMO

Pulmonary infection with Burkholderia cepacia complex in cystic fibrosis (CF) patients is associated with more-rapid lung function decline and earlier death than in CF patients without this infection. In this study, we used confocal microscopy to visualize the effects of various concentrations of tobramycin, achievable with systemic and aerosolized drug administration, on mature B. cepacia complex biofilms, both in the presence and absence of CF sputum. After 24 h of growth, biofilm thickness was significantly reduced by exposure to 2,000 µg/ml of tobramycin for Burkholderia cepacia, Burkholderia multivorans, and Burkholderia vietnamiensis; 200 µg/ml of tobramycin was sufficient to reduce the thickness of Burkholderia dolosa biofilm. With a more mature 48-h biofilm, significant reductions in thickness were seen with tobramycin at concentrations of ≥100 µg/ml for all Burkholderia species. In addition, an increased ratio of dead to live cells was observed in comparison to control with tobramycin concentrations of ≥200 µg/ml for B. cepacia and B. dolosa (24 h) and ≥100 µg/ml for Burkholderia cenocepacia and B. dolosa (48 h). Although sputum significantly increased biofilm thickness, tobramycin concentrations of 1,000 µg/ml were still able to significantly reduce biofilm thickness of all B. cepacia complex species with the exception of B. vietnamiensis. In the presence of sputum, 1,000 µg/ml of tobramycin significantly increased the dead-to-live ratio only for B. multivorans compared to control. In summary, although killing is attenuated, high-dose tobramycin can effectively decrease the thickness of B. cepacia complex biofilms, even in the presence of sputum, suggesting a possible role as a suppressive therapy in CF.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Complexo Burkholderia cepacia/efeitos dos fármacos , Burkholderia/efeitos dos fármacos , Fibrose Cística/microbiologia , Tobramicina/farmacologia , Biofilmes/crescimento & desenvolvimento , Burkholderia/crescimento & desenvolvimento , Burkholderia/ultraestrutura , Complexo Burkholderia cepacia/crescimento & desenvolvimento , Complexo Burkholderia cepacia/ultraestrutura , Criança , Fibrose Cística/patologia , Relação Dose-Resposta a Droga , Humanos , Testes de Sensibilidade Microbiana , Microscopia Confocal , Especificidade da Espécie , Escarro/química , Escarro/microbiologia
6.
Antimicrob Agents Chemother ; 59(1): 711-3, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25348526

RESUMO

Burkholderia cepacia complex and Stenotrophomonas maltophilia infections are associated with poor clinical outcomes in persons with cystic fibrosis (CF). The MIC50 based on planktonic growth and the biofilm concentration at which 50% of the isolates tested are inhibited (BIC50) of tobramycin were measured for 180 B. cepacia complex and 101 S. maltophilia CF isolates and were 100 µg/ml for both species. New inhalation devices that deliver high tobramycin levels to the lung may be able to exceed these MICs.


Assuntos
Antibacterianos/uso terapêutico , Complexo Burkholderia cepacia/efeitos dos fármacos , Fibrose Cística/microbiologia , Stenotrophomonas maltophilia/efeitos dos fármacos , Tobramicina/uso terapêutico , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Infecções por Burkholderia/tratamento farmacológico , Infecções por Burkholderia/etiologia , Infecções por Burkholderia/microbiologia , Fibrose Cística/complicações , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/etiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Tobramicina/farmacologia
7.
Int J Syst Evol Microbiol ; 65(7): 2265-2271, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25872960

RESUMO

Nine Burkholderia cepacia complex (Bcc) bacteria were isolated during environmental surveys for the ecological niche of Burkholderia pseudomallei, the aetiological agent of melioidosis, in the Northern Territory of Australia. They represented two multi-locus sequence analysis-based clusters, referred to as Bcc B and Bcc L. Three additional environmental and clinical Bcc B isolates were identified upon deposition of the sequences in the PubMLST database. Analysis of the concatenated nucleotide sequence divergence levels within both groups (1.4 and 1.9%, respectively) and towards established Bcc species (4.0 and 3.9%, respectively) demonstrated that the two taxa represented novel Bcc species. All 12 isolates were further characterized using 16S rRNA and recA gene sequence analysis, RAPD analysis, DNA base content determination, fatty acid methyl ester analysis and biochemical profiling. Analysis of recA gene sequences revealed a remarkable diversity within each of these taxa, but, together, the results supported the affiliation of the two taxa to the Bcc. Bcc B strains can be differentiated from most other Bcc members by the assimilation of maltose. Bcc L strains can be differentiated from other Bcc members by the absence of assimilation of N-acetylglucosamine. The names Burkholderia stagnalis sp. nov. with type strain LMG 28156(T) ( = CCUG 65686(T)) and Burkholderia territorii sp. nov. with type strain LMG 28158(T) ( = CCUG 65687(T)) are proposed for Bcc B and Bcc L bacteria, respectively.


Assuntos
Complexo Burkholderia cepacia/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , Complexo Burkholderia cepacia/genética , Complexo Burkholderia cepacia/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Humanos , Dados de Sequência Molecular , Tipagem de Sequências Multilocus , Northern Territory , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Análise de Sequência de DNA , Microbiologia do Solo , Escarro/microbiologia , Microbiologia da Água
8.
J Biol Chem ; 288(21): 15098-109, 2013 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-23572517

RESUMO

Metabolic footprinting of supernatants has been proposed as a tool for assigning gene function. We used NMR spectroscopy to measure the exometabolome of 86 single-gene transposon insertion mutant strains (mutants from central carbon metabolism and regulatory mutants) of the opportunistic pathogen Pseudomonas aeruginosa, grown on a medium designed to represent the nutritional content of cystic fibrosis sputum. Functionally related genes had similar metabolic profiles. E.g. for two-component system mutants, the cognate response regulator and sensor kinase genes clustered tightly together. Some strains had metabolic phenotypes (metabotypes) that could be related to the known gene function. E.g. pyruvate dehydrogenase mutants accumulated large amounts of pyruvate in the medium. In other cases, the metabolic phenotypes were not easily interpretable. The rpoN mutant, which lacks the alternative σ factor RpoN (σ(54)), accumulated high levels of gluconate in the medium. In addition, endometabolome profiling of intracellular metabolites identified a number of systemic metabolic changes. We linked this to indirect regulation of the catabolite repression protein Crc via the non-coding RNA crcZ and found that a crcZ (but not crc) mutant also shared the high-gluconate phenotype. We profiled an additional set of relevant metabolic enzymes and transporters, including Crc targets, and showed that the Crc-regulated edd mutant (gluconate-6-phosphate dehydratase) had similar gluconate levels as the rpoN mutant. Finally, a set of clinical isolates showed patient- and random amplification of polymorphic DNA (RAPD) type-specific differences in gluconate production, which were associated significantly with resistance across four antibiotics (tobramycin, ciprofloxacin, aztreonam, and imipenem), indicating that this has potential clinical relevance.


Assuntos
Proteínas de Bactérias/metabolismo , Fibrose Cística/microbiologia , Gluconatos/metabolismo , Metaboloma , Infecções por Pseudomonas/metabolismo , Pseudomonas aeruginosa/metabolismo , Proteínas de Bactérias/genética , Fibrose Cística/patologia , Farmacorresistência Bacteriana/fisiologia , Feminino , Humanos , Masculino , Infecções por Pseudomonas/genética , Infecções por Pseudomonas/patologia , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Técnica de Amplificação ao Acaso de DNA Polimórfico
9.
Antimicrob Agents Chemother ; 58(9): 5211-9, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24957830

RESUMO

Burkholderia cepacia complex (Bcc) pulmonary infections in people living with cystic fibrosis (CF) are difficult to treat because of the extreme intrinsic resistance of most isolates to a broad range of antimicrobials. Fosmidomycin is an antibacterial and antiparasitic agent that disrupts the isoprenoid biosynthesis pathway, a precursor to hopanoid biosynthesis. Hopanoids are involved in membrane stability and contribute to polymyxin resistance in Bcc bacteria. Checkerboard MIC assays determined that although isolates of the Bcc species B. multivorans were highly resistant to treatment with fosmidomycin or colistin (polymyxin E), antimicrobial synergy was observed in certain isolates when the antimicrobials were used in combination. Treatment with fosmidomycin decreased the MIC of colistin for isolates as much as 64-fold to as low as 8 µg/ml, a concentration achievable with colistin inhalation therapy. A liquid chromatography-tandem mass spectrometry technique was developed for the accurate quantitative determination of underivatized hopanoids in total lipid extracts, and bacteriohopanetetrol cyclitol ether (BHT-CE) was found to be the dominant hopanoid made by B. multivorans. The amount of BHT-CE made was significantly reduced upon fosmidomycin treatment of the bacteria. Uptake assays with 1-N-phenylnaphthylamine were used to determine that dual treatment with fosmidomycin and colistin increases membrane permeability, while binding assays with boron-dipyrromethene-conjugated polymyxin B illustrated that the addition of fosmidomycin had no impact on polymyxin binding. This work indicates that pharmacological suppression of membrane hopanoids with fosmidomycin treatment can increase the susceptibility of certain clinical B. multivorans isolates to colistin, an agent currently in use to treat pulmonary infections in CF patients.


Assuntos
Antibacterianos/farmacologia , Burkholderia/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Colistina/farmacologia , Fosfomicina/análogos & derivados , Triterpenos Pentacíclicos/análise , Membrana Celular/química , Sinergismo Farmacológico , Fosfomicina/farmacologia , Testes de Sensibilidade Microbiana , Triterpenos Pentacíclicos/antagonistas & inibidores
10.
Sci Rep ; 14(1): 8926, 2024 04 18.
Artigo em Inglês | MEDLINE | ID: mdl-38637558

RESUMO

To evaluate immune responses to COVID-19 vaccines in adults aged 50 years and older, spike protein (S)-specific antibody concentration, avidity, and function (via angiotensin-converting enzyme 2 (ACE2) inhibition surrogate neutralization and antibody dependent cellular phagocytosis (ADCP)), as well as S-specific T cells were quantified via activation induced marker (AIM) assay in response to two-dose series. Eighty-four adults were vaccinated with either: mRNA/mRNA (mRNA-1273 and/or BNT162b2); ChAdOx1-S/mRNA; or ChAdOx1-S/ChAdOx1-S. Anti-S IgG concentrations, ADCP scores and ACE2 inhibiting antibody concentrations were highest at one-month post-second dose and declined by four-months post-second dose for all groups. mRNA/mRNA and ChAdOx1-S/mRNA schedules had significantly higher antibody responses than ChAdOx1-S/ChAdOx1-S. CD8+ T-cell responses one-month post-second dose were associated with increased ACE2 surrogate neutralization. Antibody avidity (total relative avidity index) did not change between one-month and four-months post-second dose and did not significantly differ between groups by four-months post-second dose. In determining COVID-19 correlates of protection, a measure that considers both antibody concentration and avidity should be considered.


Assuntos
Vacinas contra COVID-19 , COVID-19 , Adulto , Humanos , Pessoa de Meia-Idade , Idoso , Enzima de Conversão de Angiotensina 2 , Vacina BNT162 , Estudos Prospectivos , COVID-19/prevenção & controle , Canadá/epidemiologia , Anticorpos , ChAdOx1 nCoV-19 , RNA Mensageiro , Anticorpos Antivirais , Vacinação
11.
Microorganisms ; 11(1)2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36677442

RESUMO

There is an impending crisis in healthcare brought about by a new era of untreatable infections caused by bacteria resistant to all available antibiotics. Thus, there is an urgent need to identify novel antimicrobial agents to counter the continuing threat posed by formerly treatable infections. We previously reported that a natural mineral clay known as Kisameet clay (KC) is a potent inhibitor of the organisms responsible for acute infections. Chronic bacterial infections present another major challenge to treatment by antimicrobials, due to their prolonged nature, which results in repeated exposure to antibiotics and a constant selection for antimicrobial resistance. A prime example is bacteria belonging to the Burkholderia cepacia complex (Bcc), which particularly causes some of the most serious chronic lung infections in patients with cystic fibrosis (CF) associated with unpredictable clinical outcomes, poor prognosis, and high mortality rates. Eradication of these organisms from CF patients with limited effective antimicrobial options is a major challenge. Novel therapeutic approaches are urgently required. Here, we report the in vitro antibacterial activity of KC aqueous suspensions (1-10% w/v) and its aqueous extract (L100) against a collection of extensively and multi-drug resistant clinical isolates of Bcc, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia isolated from patients with CF. These findings present a potential novel therapy for further investigation in the clinic.

12.
Infect Genet Evol ; 113: 105484, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37531976

RESUMO

OBJECTIVES: Clustering pathogen sequence data is a common practice in epidemiology to gain insights into the genetic diversity and evolutionary relationships among pathogens. We can find groups of cases with a shared transmission history and common origin, as well as identifying transmission hotspots. Motivated by the experience of clustering SARS-CoV-2 cases using whole genome sequence data during the COVID-19 pandemic to aid with public health investigation, we investigated how differences in epidemiology and sampling can influence the composition of clusters that are identified. METHODS: We performed genomic clustering on simulated SARS-CoV-2 outbreaks produced with different transmission rates and levels of genomic diversity, along with varying the proportion of cases sampled. RESULTS: In single outbreaks with a low transmission rate, decreasing the sampling fraction resulted in multiple, separate clusters being identified where intermediate cases in transmission chains are missed. Outbreaks simulated with a high transmission rate were more robust to changes in the sampling fraction and largely resulted in a single cluster that included all sampled outbreak cases. When considering multiple outbreaks in a sampled jurisdiction seeded by different introductions, low genomic diversity between introduced cases caused outbreaks to be merged into large clusters. If the transmission and sampling fraction, and diversity between introductions was low, a combination of the spurious break-up of outbreaks and the linking of closely related cases in different outbreaks resulted in clusters that may appear informative, but these did not reflect the true underlying population structure. Conversely, genomic clusters matched the true population structure when there was relatively high diversity between introductions and a high transmission rate. CONCLUSION: Differences in epidemiology and sampling can impact our ability to identify genomic clusters that describe the underlying population structure. These findings can help to guide recommendations for the use of pathogen clustering in public health investigations.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/epidemiologia , Pandemias , Surtos de Doenças , Genômica , Análise por Conglomerados
13.
Am J Respir Crit Care Med ; 183(1): 67-72, 2011 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-20709823

RESUMO

RATIONALE: infection with Burkholderia cepacia complex (BCC) bacteria in cystic fibrosis (CF) is associated with an unpredictable rate of pulmonary decline. Some BCC, but not others, elaborate copious mucoid exopolysaccharide, endowing them with a gross mucoid phenotype, the clinical significance of which has not been described. OBJECTIVES: to determine whether there was a correlation between bacterial mucoid phenotype, as assessed in a semiquantitative manner from plate culture, and severity of disease as assessed by the rate of decline in lung function. METHODS: we performed a retrospective clinical review of 100 patients with CF attending the Vancouver clinics between 1981 and 2007 and analyzed the rate of lung function decline (% predicted FEV(1)). MEASUREMENTS AND MAIN RESULTS: patients infected exclusively with nonmucoid BCC had a more rapid decline in lung function (annual FEV(1) change, -8.51 ± 2.41%) than those infected with mucoid bacteria (-3.01 ± 1.09%; P < 0.05). Linear mixed-effects data modeling revealed a statistically significant inverse association between semiquantitative mucoid exopolysaccharide production and rate of decline of lung function. In vitro incubation of BCC with ceftazidime and ciprofloxacin but not meropenem caused conversion of BCC from mucoid to nonmucoid. CONCLUSIONS: our data suggest an inverse correlation between the quantity of mucoid exopolysaccharide production by BCC bacteria and rate of decline in CF lung function. Certain antibiotics may induce a change in bacterial morphology that enhances their virulence. A simple in vitro test of bacterial mucoidy may be useful in predicting the rate of decline of respiratory function in CF.


Assuntos
Infecções por Burkholderia/microbiologia , Complexo Burkholderia cepacia/patogenicidade , Fibrose Cística/microbiologia , Pulmão/microbiologia , Muco/microbiologia , Adolescente , Adulto , Infecções por Burkholderia/complicações , Infecções por Burkholderia/fisiopatologia , Complexo Burkholderia cepacia/isolamento & purificação , Fibrose Cística/complicações , Fibrose Cística/fisiopatologia , Progressão da Doença , Feminino , Humanos , Masculino , Testes de Função Respiratória , Estudos Retrospectivos , Índice de Gravidade de Doença , Virulência , Adulto Jovem
14.
J Infect Dis ; 203(3): 383-92, 2011 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-21208930

RESUMO

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 activity. All but one isolate produced N-acyl-homoserine lactones. The B. cenocepacia N-acyl-homoserine lactone-negative isolate contained mutations in cepR and cciR. Growth competition assays were performed that compared B. cenocepacia clinical and laboratory defined wild-type and QS mutants. Survival of the laboratory wild-type and QS mutants varied, dependent on the mutation. The clinical wild-type isolate demonstrated a growth advantage over its QS mutant. These data suggest that there is a selective advantage for strains with QS systems and that QS mutations do not occur at a high frequency in B. cepacia complex isolates.


Assuntos
Infecções por Burkholderia/microbiologia , Complexo Burkholderia cepacia/fisiologia , Fibrose Cística/complicações , Percepção de Quorum/fisiologia , Acil-Butirolactonas/metabolismo , Infecções por Burkholderia/complicações , Complexo Burkholderia cepacia/genética , Complexo Burkholderia cepacia/metabolismo , Regulação Bacteriana da Expressão Gênica , Humanos , Mutação
15.
J Immunol Methods ; 507: 113306, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35705121

RESUMO

We developed a salivary assay for the detection of naturally acquired IgA antibody against Haemophilus influenzae type a (Hia) capsular polysaccharide in healthy Indigenous children from Northwestern Ontario, Canada. Hia-specific IgA antibody was detected in the saliva of 93% of Indigenous children aged 2-7 years.


Assuntos
Infecções por Haemophilus , Anticorpos Antibacterianos , Criança , Infecções por Haemophilus/diagnóstico , Haemophilus influenzae , Humanos , Imunoglobulina A , Lactente , Saliva
16.
Antimicrob Agents Chemother ; 55(5): 2256-64, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21321142

RESUMO

Burkholderia cepacia complex (BCC) bacteria are opportunistic pathogens that can cause severe disease in cystic fibrosis (CF) patients and other immunocompromised individuals and are typically multidrug resistant. Here we observed that unlike other BCC species, most environmental and clinical Burkholderia vietnamiensis isolates were intrinsically susceptible to aminoglycosides but not to cationic antimicrobial peptides or polymyxin B. Furthermore, strains acquired aminoglycoside resistance during chronic CF infection, a phenomenon that could be induced under tobramycin or azithromycin pressure in vitro. In comparing susceptible and resistant B. vietnamiensis isolates, no gross differences in lipopolysaccharide structure were observed, all had lipid A-associated 4-amino-4-deoxy-L-arabinose residues, and all were resistant to the permeabilizing effects of aminoglycosides, a measure of drug entry via self-promoted uptake. However, susceptible isolates accumulated 5 to 6 times more gentamicin than a resistant isolate, and aminoglycoside susceptibility increased in the presence of an efflux pump inhibitor. B. vietnamiensis is therefore unusual among BCC bacteria in its susceptibility to aminoglycosides and capacity to acquire resistance. Aminoglycoside resistance appears to be due to decreased cellular accumulation as a result of active efflux.


Assuntos
Aminoglicosídeos/farmacologia , Antibacterianos/farmacologia , Burkholderia/efeitos dos fármacos , Gentamicinas/farmacologia , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Polimixina B/farmacologia
17.
Microbiology (Reading) ; 157(Pt 11): 3124-3137, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21835880

RESUMO

Burkholderia cepacia complex (Bcc) bacteria are opportunistic pathogens infecting hosts such as cystic fibrosis (CF) patients. Long-term Bcc infection of CF patients' 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 trait development during lung infection. Expression profiling of mucoid D2095 and non-mucoid D2214 isolates revealed decreased expression of genes encoding products related to virulence-associated traits and metabolism in D2214. Furthermore, D2214 showed no exopolysaccharide production, lower motility and chemotaxis, and more biofilm formation, particularly under microaerophilic conditions, than the clonal mucoid isolate D2095. When Galleria mellonella was used as acute infection model, D2214 at a cell number of approximately 7 × 106 c.f.u. caused a higher survival rate than D2095, although 6 days post-infection most of the larvae were dead. Infection with the same number of cells by mucoid D2095 caused larval death by day 4. The decreased expression of genes involved in carbon and nitrogen metabolism may reflect lower metabolic needs of D2214 caused by lack of exopolysaccharide, but also by the attenuation of pathways not required for survival. As a result, D2214 showed higher survival than D2095 in minimal medium for 28 days under aerobic conditions. Overall, adaptation during Bcc chronic lung infections gave rise to genotypic and phenotypic variation among isolates, contributing to their fitness while maintaining their capacity for survival in this opportunistic human niche.


Assuntos
Biofilmes , Infecções por Burkholderia/microbiologia , Complexo Burkholderia cepacia/patogenicidade , Fibrose Cística/complicações , Infecções Respiratórias/microbiologia , Adaptação Fisiológica , Complexo Burkholderia cepacia/genética , Complexo Burkholderia cepacia/isolamento & purificação , Complexo Burkholderia cepacia/metabolismo , DNA Bacteriano/genética , Regulação Bacteriana da Expressão Gênica , Genótipo , Humanos , Pulmão/microbiologia , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Infecções Oportunistas/microbiologia , Fenótipo , RNA Bacteriano/genética , Transcriptoma , Virulência
19.
J Infect Dis ; 202(5): 770-81, 2010 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-20670172

RESUMO

Bacteria of the Burkholderia cepacia complex (BCC) are associated with severe infection in cystic fibrosis. Recent evidence shows that the mucoid phenotype is common in BCC bacteria; however, during chronic infection, transitions from the mucoid to nonmucoid morphology have been shown to take place. Here we use RNA microarray and proteomic isobaric tagging relative and absolute quantitation technologies to gain insight into a pair of mucoid and nonmucoid isolates of B. cenocepacia obtained from a chronically infected patient with cystic fibrosis in the year prior to her death. During chronic infection, the mucoid isolate lost the B. cepacia epidemic strain marker and acquired a mutation in the cepR gene. In the nonmucoid isolate, we observed overexpression at both the RNA and protein level of several described putative virulence factors, including a nematocidal protein AidA and the oxidative stress response protein AhpC. We show that this translates into increased resistance to oxidative stress in the nonmucoid isolate, a key microbial determinant for resistance against phagocytic cell killing. These data illuminate the biological differences between mucoid and nonmucoid BCC bacteria, provide targets for elucidating the genetic control of exopolysaccharide production in the BCC, and highlight that chronic infection can produce both genetically and phenotypically distinct microbial variants in the cystic fibrosis lung.


Assuntos
Infecções por Burkholderia/fisiopatologia , Complexo Burkholderia cepacia/fisiologia , Complexo Burkholderia cepacia/patogenicidade , Fibrose Cística/complicações , Perfilação da Expressão Gênica , Proteômica , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Infecções por Burkholderia/microbiologia , Complexo Burkholderia cepacia/crescimento & desenvolvimento , Complexo Burkholderia cepacia/isolamento & purificação , Fibrose Cística/microbiologia , Fibrose Cística/fisiopatologia , Regulação Bacteriana da Expressão Gênica , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Virulência , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
20.
Microbiol Spectr ; 9(1): e0012721, 2021 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-34132614

RESUMO

The Burkholderia cepacia complex (Bcc) comprises several species of closely related, versatile bacteria. Some Bcc strains produce 4-hydroxy-3-methyl-2-alkylquinolines (HMAQs), analogous to the 4-hydroxy-2-alkylquinolines of Pseudomonas aeruginosa. Using in silico analyses, we previously estimated that the hmqABCDEFG operon, which encodes enzymes involved in the biosynthesis of HMAQs, is carried by about one-third of Bcc strains, with considerable inter- and intraspecies variability. In the present study, we investigated by PCR, using consensus primers, the distribution of hmqABCDEFG in a collection of 312 Bcc strains (222 of clinical and 90 of environmental origins) belonging to 18 Bcc species. We confirmed that this operon is not distributed evenly among Bcc species. Among the 30% of strains bearing the hmqABCDEFG operon, we found that 92% of environmental isolates and 82% of clinically isolated Bcc strains produce levels of HMAQs detectable by liquid chromatography-mass spectrometry in at least one of the tested culture conditions. Among the hmqABCDEFG-positive but HMAQ-negative strains, none expressed the hmqA gene under the specified culture conditions. Interestingly, the hmqABCDEFG operon is more prevalent among plant root environment species (e.g., Burkholderia ambifaria and Burkholderia cepacia) and absent in species commonly found in chronically colonized individuals with cystic fibrosis (e.g., Burkholderia cenocepacia and Burkholderia multivorans), suggesting a role for the Hmq system in niche adaptation. We investigated the impact of the Hmq system on plant growth promotion and found that Pisum sativum root development by B. ambifaria required a functional HMAQ system. IMPORTANCE Environmental bacteria belonging to the various closely related species forming the Burkholderia cepacia complex (Bcc) can infect plants and animals, including humans. Their pathogenicity is regulated by intercellular communication, or quorum sensing, allowing them to collaborate instead of acting individually. Bcc organisms generally exploit interacting quorum sensing systems based on N-acyl-homoserine lactones as signaling molecules. Several Bcc strains also carry an hmqABCDEFG operon responsible for the biosynthesis of 4-hydroxy-3-methyl-2-alkylquinolines (HMAQs), molecules analogous to the Pseudomonas quinolone signal (PQS) system of P. aeruginosa. Our finding that the prevalences of the Hmq system and HMAQ production are very different between various Bcc species suggests a key role in niche adaptation or pathogenicity. This is supported by a significant reduction in plant growth promotion in the absence of HMAQ production for a beneficial Bcc strain.


Assuntos
Proteínas de Bactérias/genética , Infecções por Burkholderia/microbiologia , Complexo Burkholderia cepacia/metabolismo , Óperon , Raízes de Plantas/microbiologia , Quinolinas/metabolismo , Proteínas de Bactérias/metabolismo , Complexo Burkholderia cepacia/classificação , Complexo Burkholderia cepacia/genética , Complexo Burkholderia cepacia/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Fibrose Cística/microbiologia , Humanos , Espectrometria de Massas , Raízes de Plantas/crescimento & desenvolvimento , Quinolinas/química
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