RESUMO
Thymol, carvacrol, and trans-cinnamaldehyde are essential oil (EO) compounds with broad-spectrum antimicrobial activities against foodborne pathogens, including Escherichia coli O157:H7. However, little is known regarding direct resistance and cross-resistance development in E. coli O157:H7 after adaptation to sublethal levels of these compounds, and information is scarce on microbial adaptive responses at a molecular level. The present study demonstrated that E. coli O157:H7 was able to grow in the presence of sublethal thymol (1/2T), carvacrol (1/2C), or trans-cinnamaldehyde (1/2TC), displaying an extended lag phase duration and a lower maximum growth rate. EO-adapted cells developed direct resistance against lethal EO treatments and cross-resistance against heat (58°C) and oxidative (50 mM H2O2) stresses. However, no induction of acid resistance (simulated gastric fluid, pH 1.5) was observed. RNA sequencing revealed a large number (310 to 338) of differentially expressed (adjusted P value [Padj ], <0.05; fold change, ≥5) genes in 1/2T and 1/2C cells, while 1/2TC cells only showed 27 genes with altered expression. In accordance with resistance phenotypes, the genes related to membrane, heat, and oxidative stress responses and genes related to iron uptake and metabolism were upregulated. Conversely, virulence genes associated with motility, biofilm formation, and efflux pumps were repressed. This study demonstrated the development of direct resistance and cross-resistance and characterized whole-genome transcriptional responses in E. coli O157:H7 adapted to sublethal thymol, carvacrol, or trans-cinnamaldehyde. The data suggested that caution should be exercised when using EO compounds as food antimicrobials, due to the potential stress resistance development in E. coli O157:H7.IMPORTANCE The present study was designed to understand transcriptomic changes and the potential development of direct and cross-resistance in essential oil (EO)-adapted Escherichia coli O157:H7. The results demonstrated altered growth behaviors of E. coli O157:H7 during adaptation in sublethal thymol, carvacrol, and trans-cinnamaldehyde. Generally, EO-adapted bacteria showed enhanced resistance against subsequent lethal EO, heat, and oxidative stresses, with no induction of acid resistance in simulated gastric fluid. A transcriptomic analysis revealed the upregulation of related stress resistance genes and a downregulation of various virulence genes in EO-adapted cells. This study provides new insights into microbial EO adaptation behaviors and highlights the risk of resistance development in adapted bacteria.
Assuntos
Acroleína/análogos & derivados , Antibacterianos/metabolismo , Escherichia coli O157/fisiologia , Monoterpenos/metabolismo , Óleos Voláteis/metabolismo , Timol/metabolismo , Transcrição Gênica , Acroleína/metabolismo , Acroleína/farmacologia , Antibacterianos/farmacologia , Cimenos , Farmacorresistência Bacteriana , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/genética , Escherichia coli O157/crescimento & desenvolvimento , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Genoma Bacteriano , Temperatura Alta , Ferro/metabolismo , Monoterpenos/farmacologia , Óleos Voláteis/farmacologia , Estresse Fisiológico/efeitos dos fármacos , Timol/farmacologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Nucleotide substitutions in codons 67 and 119 of HLA-B*15:02:01:01 result in a novel allele, HLA-B*15:404.
Assuntos
Medula Óssea , Antígenos HLA-B , Alelos , Sequência de Bases , Éxons/genética , Antígenos HLA-B/genética , Teste de Histocompatibilidade , Humanos , Análise de Sequência de DNA , Singapura , Doadores de TecidosRESUMO
Nucleotide substitutions in exon 2 of DRB1*14:54:01:01 result in the novel DRB1*14:119 allele.
Assuntos
Medula Óssea , Alelos , Sequência de Bases , Cadeias HLA-DRB1/genética , Teste de Histocompatibilidade , Humanos , Análise de Sequência de DNA , SingapuraRESUMO
Long-range material transport is essential to maintain the physiological functions of multicellular organisms such as animals and plants. By contrast, material transport in bacteria is often short-ranged and limited by diffusion. Here, we report a unique form of actively regulated long-range directed material transport in structured bacterial communities. Using Pseudomonas aeruginosa colonies as a model system, we discover that a large-scale and temporally evolving open-channel system spontaneously develops in the colony via shear-induced banding. Fluid flows in the open channels support high-speed (up to 450 µm/s) transport of cells and outer membrane vesicles over centimeters, and help to eradicate colonies of a competing species Staphylococcus aureus. The open channels are reminiscent of human-made canals for cargo transport, and the channel flows are driven by interfacial tension mediated by cell-secreted biosurfactants. The spatial-temporal dynamics of fluid flows in the open channels are qualitatively described by flow profile measurement and mathematical modeling. Our findings demonstrate that mechanochemical coupling between interfacial force and biosurfactant kinetics can coordinate large-scale material transport in primitive life forms, suggesting a new principle to engineer self-organized microbial communities.
Assuntos
Microbiota , Infecções Estafilocócicas , Animais , Humanos , Pseudomonas aeruginosa/fisiologia , Staphylococcus aureus/fisiologia , Infecções Estafilocócicas/microbiologia , BactériasRESUMO
One nucleotide substitution in codon 108 of HLA-C*03:03:01:01 results in a novel allele, HLA-C*03:88.
Assuntos
Medula Óssea , Antígenos HLA-C , Alelos , Sequência de Bases , Éxons/genética , Antígenos HLA-C/genética , Teste de Histocompatibilidade , Humanos , Análise de Sequência de DNA , Doadores de TecidosRESUMO
One nucleotide substitution in codon 120 of HLA-B*18:01:01:01 results in a novel allele, HLA-B*18:116.
Assuntos
Medula Óssea , Antígenos HLA-B , Alelos , Sequência de Bases , Éxons/genética , Antígenos HLA-B/genética , Humanos , Análise de Sequência de DNA , Doadores de TecidosRESUMO
One nucleotide substitution in codon 95 of HLA-C*07:04:01:01 results in a novel allele, HLA-C*07:199:01.
Assuntos
Antígenos HLA-C , Transplante de Células-Tronco Hematopoéticas , Alelos , Sequência de Bases , Éxons/genética , Antígenos HLA-C/genética , Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Análise de Sequência de DNA , Doadores não RelacionadosRESUMO
One nucleotide substitution in Codon 58 of HLA-C*06:02:01:01 results in a novel allele, HLA-C*06:195.
Assuntos
Antígenos HLA-C , Transplante de Células-Tronco Hematopoéticas , Alelos , Sequência de Bases , Éxons/genética , Antígenos HLA-C/genética , Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Análise de Sequência de DNA , TaiwanRESUMO
Estrogen, a major female sex steroid hormone, has been shown to promote the selection of mucoid Pseudomonas aeruginosa in the airways of patients with chronic respiratory diseases, including cystic fibrosis. This results in long-term persistence, poorer clinical outcomes, and limited therapeutic options. In this study, we demonstrate that at physiological concentrations, sex steroids, including testosterone and estriol, induce membrane stress responses in P. aeruginosa This is characterized by increased virulence and consequent inflammation and release of proinflammatory outer membrane vesicles promoting in vivo persistence of the bacteria. The steroid-induced P. aeruginosa response correlates with the molecular polarity of the hormones and membrane fluidic properties of the bacteria. This novel mechanism of interaction between sex steroids and P. aeruginosa explicates the reported increased disease severity observed in females with cystic fibrosis and provides evidence for the therapeutic potential of the modulation of sex steroids to achieve better clinical outcomes in patients with hormone-responsive strains.IMPORTANCE Molecular mechanisms by which sex steroids interact with P. aeruginosa to modulate its virulence have yet to be reported. Our work provides the first characterization of a steroid-induced membrane stress mechanism promoting P. aeruginosa virulence, which includes the release of proinflammatory outer membrane vesicles, resulting in inflammation, host tissue damage, and reduced bacterial clearance. We further demonstrate that at nanomolar (physiological) concentrations, male and female sex steroids promote virulence in clinical strains of P. aeruginosa based on their dynamic membrane fluidic properties. This work provides, for the first-time, mechanistic insight to better understand and predict the P. aeruginosa related response to sex steroids and explain the interindividual patient variability observed in respiratory diseases such as cystic fibrosis that are complicated by gender differences and chronic P. aeruginosa infection.
Assuntos
Membrana Externa Bacteriana/efeitos dos fármacos , Fibrose Cística/complicações , Hormônios Esteroides Gonadais/metabolismo , Pseudomonas aeruginosa/patogenicidade , Estresse Fisiológico/efeitos dos fármacos , Alginatos/metabolismo , Animais , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Fibrose Cística/microbiologia , Estradiol/química , Estradiol/farmacologia , Feminino , Hormônios Esteroides Gonadais/farmacologia , Humanos , Inflamação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pseudomonas aeruginosa/genética , Fatores Sexuais , Testosterona/química , Testosterona/farmacologia , VirulênciaRESUMO
Mixed-species biofilms display a number of emergent properties, including enhanced antimicrobial tolerance and communal metabolism. These properties may depend on interspecies relationships and the structure of the biofilm. However, the contribution of specific matrix components to emergent properties of mixed-species biofilms remains poorly understood. Using a dual-species biofilm community formed by the opportunistic pathogens Pseudomonas aeruginosa and Staphylococcus aureus, we found that whilst neither Pel nor Psl polysaccharides, produced by P. aeruginosa, affect relative species abundance in mature P. aeruginosa and S. aureus biofilms, Psl production is associated with increased P. aeruginosa abundance and reduced S. aureus aggregation in the early stages of biofilm formation. Our data suggest that the competitive effect of Psl is not associated with its structural role in cross-linking the matrix and adhering to P. aeruginosa cells but is instead mediated through the activation of the diguanylate cyclase SiaD. This regulatory control was also found to be independent of the siderophore pyoverdine and Pseudomonas quinolone signal, which have previously been proposed to reduce S. aureus viability by inducing lactic acid fermentation-based growth. In contrast to the effect mediated by Psl, Pel reduced the effective crosslinking of the biofilm matrix and facilitated superdiffusivity in microcolony regions. These changes in matrix cross-linking enhance biofilm surface spreading and expansion of microcolonies in the later stages of biofilm development, improving overall dual-species biofilm growth and increasing biovolume severalfold. Thus, the biofilm matrix and regulators associated with matrix production play essential roles in mixed-species biofilm interactions.IMPORTANCE Bacteria in natural and engineered environments form biofilms that include many different species. Microorganisms rely on a number of different strategies to manage social interactions with other species and to access resources, build biofilm consortia, and optimize growth. For example, Pseudomonasaeruginosa and Staphylococcus aureus are biofilm-forming bacteria that coinfect the lungs of cystic fibrosis patients and diabetic and chronic wounds. P. aeruginosa is known to antagonize S. aureus growth. However, many of the factors responsible for mixed-species interactions and outcomes such as infections are poorly understood. Biofilm bacteria are encased in a self-produced extracellular matrix that facilitates interspecies behavior and biofilm development. In this study, we examined the poorly understood roles of the major matrix biopolymers and their regulators in mixed-species biofilm interactions and development.