RESUMEN
Prokaryotic cell transcriptomics has been limited to mixed or sub-population dynamics and individual cells within heterogeneous populations, which has hampered further understanding of spatiotemporal and stage-specific processes of prokaryotic cells within complex environments. Here we develop a 'TRANSITomic' approach to profile transcriptomes of single Burkholderia pseudomallei cells as they transit through host cell infection at defined stages, yielding pathophysiological insights. We find that B. pseudomallei transits through host cells during infection in three observable stages: vacuole entry; cytoplasmic escape and replication; and membrane protrusion, promoting cell-to-cell spread. The B. pseudomallei 'TRANSITome' reveals dynamic gene-expression flux during transit in host cells and identifies genes that are required for pathogenesis. We find several hypothetical proteins and assign them to virulence mechanisms, including attachment, cytoskeletal modulation, and autophagy evasion. The B. pseudomallei 'TRANSITome' provides prokaryotic single-cell transcriptomics information enabling high-resolution understanding of host-pathogen interactions.
Asunto(s)
Proteínas Bacterianas/genética , Burkholderia pseudomallei/genética , Biología Computacional/métodos , Perfilación de la Expresión Génica/métodos , Genes Bacterianos/genética , Factores de Virulencia/genética , Animales , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/patogenicidad , Línea Celular Tumoral , Membrana Celular/microbiología , Citoplasma/microbiología , Células HEK293 , Interacciones Huésped-Patógeno , Humanos , Melioidosis/microbiología , Ratones , Ratones Endogámicos BALB C , Células RAW 264.7 , Análisis de la Célula Individual/métodos , Vacuolas/microbiología , Virulencia/genéticaRESUMEN
Burkholderia pseudomallei is the causative agent of the tropical disease melioidosis. Its genome encodes an arsenal of virulence factors that allow it, when required, to switch from a soil dwelling bacterium to a deadly intracellular pathogen. With a high intrinsic resistance to antibiotics and the ability to overcome challenges from the host immune system, there is an increasing requirement for new antibiotics and a greater understanding into the molecular mechanisms of B. pseudomallei virulence and dormancy. The peptidoglycan remodeling enzymes, lytic transglycosylases (Ltgs) are potential targets for such new antibiotics. Ltgs cleave the glycosidic bonds within bacterial peptidoglycan allowing for the insertion of peptidoglycan precursors during cell growth and division, and cell membrane spanning structures such as flagella and secretion systems. Using bioinformatic analysis we have identified 8 putative Ltgs in B. pseudomallei K96243. We aimed to investigate one of these Ltgs, LtgG (BPSL3046) through the generation of deletion mutants and biochemical analysis. We have shown that LtgG is a key contributor to cellular morphology, division, motility and virulence in BALB/c mice. We have determined the crystal structure of LtgG and have identified various amino acids likely to be important in peptidoglycan binding and catalytic activity. Recombinant protein assays and complementation studies using LtgG containing a site directed mutation in aspartate 343, confirmed the essentiality of this amino acid in the function of LtgG.
Asunto(s)
Proteínas Bacterianas/metabolismo , Burkholderia pseudomallei/metabolismo , Melioidosis/microbiología , Peptidoglicano Glicosiltransferasa/metabolismo , Animales , Proteínas Bacterianas/genética , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/genética , Burkholderia pseudomallei/patogenicidad , Membrana Celular/metabolismo , Forma de la Célula , Biología Computacional , Ratones , Ratones Endogámicos BALB C , Peptidoglicano Glicosiltransferasa/genética , Virulencia/genéticaRESUMEN
Intracellular actin-based motility of the melioidosis pathogen Burkholderia pseudomallei requires the bacterial factor BimA. Located at one pole of the bacterium, BimA recruits and polymerizes cellular actin to promote bacterial motility within and between cells. Here, we describe an affinity approach coupled with mass spectrometry to identify cellular proteins recruited to BimA-expressing bacteria under conditions that promote actin polymerization. We identified a group of cellular proteins that are recruited to the B. pseudomallei surface in a BimA-dependent manner, a subset of which were independently validated with specific antisera including the ubiquitous scaffold protein Ras GTPase-activating-like protein (IQGAP1). IQGAP1 integrates several key cellular signaling pathways including those involved in actin dynamics and has been shown to be involved in the adhesion of attaching and effacing Escherichia coli to infected cells and invasion of host cells by Salmonella enterica serovar Typhimurium. Although a direct interaction between BimA and IQGAP1 could not be detected using either conventional pulldown or yeast two hybrid techniques, confocal microscopy revealed that IQGAP1 is recruited to B. pseudomallei actin tails in infected cells, and siRNA-mediated knockdown highlighted a role for this protein in controlling the length and actin density of B. pseudomallei actin tails.
Asunto(s)
Actinas/metabolismo , Burkholderia pseudomallei/química , Movimiento Celular , Proteínas Bacterianas/análisis , Proteínas Bacterianas/fisiología , Burkholderia pseudomallei/citología , Polaridad Celular , Humanos , Proteínas de Microfilamentos/metabolismo , Proteínas de Microfilamentos/fisiología , Polimerizacion , Proteínas Activadoras de ras GTPasa/metabolismo , Proteínas Activadoras de ras GTPasa/fisiologíaRESUMEN
Melioidosis sepsis, caused by Burkholderia pseudomallei, is associated with high mortality due to an overwhelming inflammatory response. Plasmacytoid dendritic cells (pDC) are potent producers of type I interferons (IFN). This study investigated whether pDC and type I IFN play a role during the early stages of B. pseudomallei infection. Human and murine pDC internalised and killed B. pseudomallei as efficiently as murine conventional DC (cDC). pDC derived from B. pseudomallei-susceptible (BALB/c) mice demonstrated poor intracellular killing and increased IFN-alpha compared to pDC derived from B. pseudomallei-resistant (C57BL/6) mice. This is the first evidence of pDC bactericidal activity against B. pseudomallei infection.
Asunto(s)
Burkholderia pseudomallei/fisiología , Células Dendríticas/inmunología , Animales , Burkholderia pseudomallei/citología , Células Dendríticas/citología , Modelos Animales de Enfermedad , Interferón-alfa , Interferón beta , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BLRESUMEN
Colony morphology variation is a characteristic of Burkholderia pseudomallei primary clinical isolates, associated with variations in expression of virulence factors. Here, we performed comparative investigations on adhesion, invasion, plaque-forming abilities and protein profiles of B. pseudomallei wild-type (WT) and a small colony variant (SCV). The percentage of SCV adherence to A549 cells was significantly higher (2.73%) than WT (1.91%). In contrast, WT was significantly more efficient (0.63%) than SCV (0.31%) in invasiveness and in inducing cellular damage. Using 2-DE and MALDI TOF/TOF, 263 and 258 protein spots were detected in WT and SCV, respectively. Comparatively, 49 proteins were differentially expressed in SCV when compared with WT. Of these, 31 proteins were up-regulated, namely, nucleoside diphosphate kinase (Ndk), phosphoglycerate kinase (Pgk), thioredoxin (TrxA), putative ferritin DPS-family DNA-binding protein (DPS) and oxidoreductase (AhpC) that are known to be involved in adhesion, intracellular survival and persistence. However, among the 18 down-regulated proteins, enolase (Eno), elongation factor (EF-Tu) and universal stress-related proteins were associated with invasion and virulence. Differences observed in these protein profiles provide ample clues to their association with the morphotypic and phenotypic characteristics of colony variants, providing additional insights into the potential association of B. pseudomallei colony morphotypes with disease pathogenesis. BIOLOGICAL SIGNIFICANCE: Comparative analyses were performed on the ability of wild-type and small colony variant of B. pseudomallei to adhere, invade and form plaques on human epithelial cells. In addition, proteomic analysis of these different colony morphotypes was also carried out. This research provides insights into the virulence and pathogenesis attributes of B. pseudomallei and contributes to better understand the pathogenesis of melioidosis.
Asunto(s)
Burkholderia pseudomallei/citología , Células Epiteliales/microbiología , Regulación Bacteriana de la Expresión Génica , Factores de Virulencia/metabolismo , Adhesión Bacteriana , Proteínas Bacterianas/metabolismo , Biomarcadores/metabolismo , Burkholderia pseudomallei/patogenicidad , Línea Celular Tumoral , Bases de Datos de Proteínas , Perfilación de la Expresión Génica , Humanos , Espectrometría de Masas , Melioidosis/microbiología , Proteoma , Proteómica , Virulencia/genéticaRESUMEN
TA (toxin-antitoxin) systems are widely distributed amongst bacteria and are associated with the formation of antibiotic tolerant (persister) cells that may have involvement in chronic and recurrent disease. We show that overexpression of the Burkholderia pseudomallei HicA toxin causes growth arrest and increases the number of persister cells tolerant to ciprofloxacin or ceftazidime. Furthermore, our data show that persistence towards ciprofloxacin or ceftazidime can be differentially modulated depending on the level of induction of HicA expression. Deleting the hicAB locus from B. pseudomallei K96243 significantly reduced persister cell frequencies following exposure to ciprofloxacin, but not ceftazidime. The structure of HicA(H24A) was solved by NMR and forms a dsRBD-like (dsRNA-binding domain-like) fold, composed of a triple-stranded ß-sheet, with two helices packed against one face. The surface of the protein is highly positively charged indicative of an RNA-binding protein and His24 and Gly22 were functionality important residues. This is the first study demonstrating a role for the HicAB system in bacterial persistence and the first structure of a HicA protein that has been experimentally characterized.
Asunto(s)
Toxinas Bacterianas/metabolismo , Burkholderia pseudomallei/metabolismo , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/genética , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/efectos de los fármacos , Burkholderia pseudomallei/genética , Ceftazidima/farmacología , Ciprofloxacina/farmacología , Clonación Molecular , Farmacorresistencia Bacteriana , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Regulación Bacteriana de la Expresión Génica/fisiología , Pruebas de Sensibilidad Microbiana , Mutación , Unión Proteica , Conformación Proteica , Pliegue de Proteína , Estructura Terciaria de Proteína , ARN BicatenarioRESUMEN
Burkholderia pseudomallei, a Gram-negative saprophytic bacterium, is the causative agent of the potentially fatal melioidosis disease in humans. In this study, environmental parameters including temperature, nutrient content, pH and the presence of glucose were shown to play a role in in vitro biofilm formation by 28 B. pseudomallei clinical isolates, including four isolates with large colony variants (LCVs) and small colony variants (SCVs) morphotypes. Enhanced biofilm formation was observed when the isolates were tested in LB medium, at 30 °C, at pH 7.2, and in the presence of as little as 2 mM glucose respectively. It was also shown that all SVCs displayed significantly greater capacity to form biofilms than the corresponding LCVs when cultured in LB at 37 °C. In addition, octanoyl-homoserine lactone (C(8)-HSL), a quorum sensing molecule, was identified by mass spectrometry analysis in bacterial isolates referred to as LCV CTH, LCV VIT, SCV TOM, SCV CTH, 1 and 3, and the presence of other AHL's with higher masses; decanoyl-homoserine lactone (C(10)-HSL) and dodecanoyl-homoserine lactone (C(12)-HSL) were also found in all tested strain in this study. Last but not least, we had successfully acquired two Bacillus sp. soil isolates, termed KW and SA respectively, which possessed strong AHLs degradation activity. Biofilm formation of B. pseudomallei isolates was significantly decreased after treated with culture supernatants of KW and SA strains, demonstrating that AHLs may play a role in B. pseudomallei biofilm formation.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Burkholderia pseudomallei/aislamiento & purificación , Burkholderia pseudomallei/fisiología , Ambiente , 4-Butirolactona/análogos & derivados , Animales , Bacillus/efectos de los fármacos , Bacillus/fisiología , Bioensayo , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/efectos de los fármacos , Caenorhabditis elegans/citología , Caenorhabditis elegans/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Cromatografía en Capa Delgada , Recuento de Colonia Microbiana , Medios de Cultivo/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/fisiología , Glucosa/farmacología , Humanos , Concentración de Iones de Hidrógeno/efectos de los fármacos , Espectrometría de Masas , Percepción de Quorum , TemperaturaRESUMEN
Colony morphology variation of Burkholderia pseudomallei is a notable feature of a proportion of primary clinical cultures from patients with melioidosis. Here, we examined the hypothesis that colony morphology switching results in phenotypic changes associated with enhanced survival under adverse conditions. We generated isogenic colony morphology types II and III from B. pseudomallei strain 153 type I, and compared their protein expression profiles using 2D gel electrophoresis. Numerous proteins were differentially expressed, the most prominent of which were flagellin, arginine deiminase (AD) and carbamate kinase (CK), which were over-expressed in isogenic types II and III compared with parental type I. AD and CK (encoded by arcA and arcC) are components of the arginine deiminase system (ADS) which facilitates acid tolerance. Reverse transcriptase PCR of arcA and arcC mRNA expression confirmed the proteomic results. Transcripts of parental type I strain 153 arcA and arcC were increased in the presence of arginine, in a low oxygen concentration and in acid. Comparison of wild type with arcA and arcC defective mutants demonstrated that the B. pseudomallei ADS was associated with survival in acid, but did not appear to play a role in intracellular survival or replication within the mouse macrophage cell line J774A.1. These data provide novel insights into proteomic alterations that occur during the complex process of morphotype switching, and lend support to the idea that this is associated with a fitness advantage in vivo.
Asunto(s)
Proteínas Bacterianas/biosíntesis , Burkholderia pseudomallei/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Animales , Proteínas Bacterianas/genética , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/genética , Humanos , Melioidosis/genética , Melioidosis/metabolismo , Mutación , Proteómica/métodos , ARN Bacteriano/biosíntesis , ARN Bacteriano/genética , Especificidad de la EspecieRESUMEN
The size and density of microbial cells determine the time that pathogens can remain airborne and thus, their potential to infect by the respiratory route. We determined the density and size distribution of Burkholderia pseudomallei cells in comparison with other Burkholderia species, including B. mallei and B. thailandensis, all prepared and analyzed under similar conditions. The observed size distribution and densities of several bacterial strains indicates that aerosolized particles consisting of one or of a few B. pseudomallei cells should be efficiently retained in the lungs, highlighting the risk of transmission of melioidosis by the respiratory route when the pathogen is present in fluids from infected patients or aerosolized from the environment.
Asunto(s)
Burkholderia pseudomallei/fisiología , Melioidosis/transmisión , Aerosoles , Burkholderia/citología , Burkholderia/fisiología , Burkholderia pseudomallei/citología , Humanos , Melioidosis/microbiología , Factores de Riesgo , Gravedad EspecíficaRESUMEN
BACKGROUND: Primary diagnostic cultures from patients with melioidosis demonstrate variation in colony morphology of the causative organism, Burkholderia pseudomallei. Variable morphology is associated with changes in the expression of a range of putative virulence factors. This study investigated the effect of B. pseudomallei colony variation on survival in the human macrophage cell line U937 and under laboratory conditions simulating conditions within the macrophage milieu. Isogenic colony morphology types II and III were generated from 5 parental type I B. pseudomallei isolates using nutritional limitation. Survival of types II and III were compared with type I for all assays. RESULTS: Morphotype was associated with survival in the presence of H2O2 and antimicrobial peptide LL-37, but not with susceptibility to acid, acidified sodium nitrite, or resistance to lysozyme, lactoferrin, human neutrophil peptide-1 or human beta defensin-2. Incubation under anaerobic conditions was a strong driver for switching of type III to an alternative morphotype. Differences were noted in the survival and replication of the three types following uptake by human macrophages, but marked strain-to strain-variability was observed. Uptake of type III alone was associated with colony morphology switching. CONCLUSIONS: Morphotype is associated with phenotypes that alter the ability of B. pseudomallei to survive in adverse environmental conditions.
Asunto(s)
Antibacterianos/farmacología , Burkholderia pseudomallei/citología , Farmacorresistencia Bacteriana , Macrófagos/microbiología , Melioidosis/microbiología , Viabilidad Microbiana , Péptidos Catiónicos Antimicrobianos , Burkholderia pseudomallei/clasificación , Burkholderia pseudomallei/efectos de los fármacos , Burkholderia pseudomallei/crecimiento & desarrollo , Burkholderia pseudomallei/aislamiento & purificación , Catelicidinas/farmacología , Humanos , Peróxido de Hidrógeno/farmacología , Viabilidad Microbiana/efectos de los fármacos , Células U937RESUMEN
BACKGROUND: Burkholderia pseudomallei, the causative agent of melioidosis, is generally considered to be acquired via inhalation of dust or water droplets from the environment. In this study, we show that infection of the nasal mucosa is potentially an important portal of entry in melioidosis. METHODS: After intranasal inoculation of mice, infection was monitored by bioluminescence imaging and by immunohistological analysis of coronal sections. The bacterial loads in organ and tissue specimens were also monitored. RESULTS: Bioluminescence imaging showed colonization and replication in the nasal cavity, including the nasal-associated lymphoid tissue (NALT). Analysis of coronal sections and immunofluorescence microscopy further demonstrated the presence of infection in the respiratory epithelium and the olfactory epithelium (including associated nerve bundles), as well as in the NALT. Of significance, the olfactory epithelium and the brain were rapidly infected before bacteria were detected in blood, and a capsule-deficient mutant infected the brain without significantly infecting blood. CONCLUSIONS: These data suggest that the olfactory nerve is the route of entry into the brain and that this route of entry may be paralleled in cases of human neurologic melioidosis. This study focuses attention on the upper respiratory tract as a portal of entry, specifically focusing on NALT as a route for the development of systemic infection via the bloodstream and on the olfactory epithelium as a direct route to the brain.
Asunto(s)
Burkholderia pseudomallei , Tejido Linfoide/microbiología , Melioidosis/microbiología , Mucosa Olfatoria/microbiología , Animales , Burkholderia pseudomallei/citología , División Celular , Femenino , Proteínas Luminiscentes , Melioidosis/patología , Ratones , Ratones Endogámicos BALB C , Nariz/anatomía & histología , Nariz/microbiología , Bulbo Olfatorio/microbiología , Células Receptoras Sensoriales/microbiologíaRESUMEN
Burkholderia pseudomallei and B. mallei are Gram-negative bacterial pathogens that cause melioidosis in humans and glanders in horses, respectively. Both bacteria are classified as category B select agents in the United States. Due to strict select-agent regulations, the number of antibiotic selection markers approved for use in these bacteria is greatly limited. Approved markers for B. pseudomallei include genes encoding resistance to kanamycin (Km), gentamicin (Gm), and zeocin (Zeo); however, wild type B. pseudomallei is intrinsically resistant to these antibiotics. Selection markers for B. mallei are limited to Km and Zeo resistance genes. Additionally, there are few well developed counter-selection markers for use in Burkholderia. The use of SacB as a counter-selection method has been of limited success due to the presence of endogenous sacBC genes in the genomes of B. pseudomallei and B. mallei. These impediments have greatly hampered the genetic manipulation of B. pseudomallei and B. mallei and currently few reliable tools for the genetic manipulation of Burkholderia exist. To expand the repertoire of genetic tools for use in Burkholderia, we developed the suicide plasmid pMo130, which allows for the compliant genetic manipulation of the select agents B. pseudomallei and B. mallei using allelic exchange. pMo130 harbors an aphA gene which allows for Km selection, the reporter gene xylE, which allows for reliable visual detection of Burkholderia transformants, and carries a modified sacB gene that allows for the resolution of co-integrants. We employed this system to generate multiple unmarked and in-frame mutants in B. pseudomallei, and one mutant in B. mallei. This vector significantly expands the number of available tools that are select-agent compliant for the genetic manipulation of B. pseudomallei and B. mallei.
Asunto(s)
Alelos , Burkholderia mallei/genética , Burkholderia pseudomallei/genética , Técnicas Genéticas , Burkholderia mallei/citología , Burkholderia pseudomallei/citología , Flagelos/genética , Prueba de Complementación Genética , Vectores Genéticos/genética , Movimiento , Plásmidos/genética , Reacción en Cadena de la Polimerasa , Eliminación de SecuenciaRESUMEN
Burkholderia pseudomallei is the causative agent of melioidosis, a tropical infection of humans and other animals. The bacterium is an intracellular pathogen that can escape from endosomes into the host cytoplasm, where it replicates and infects adjacent cells. We investigated the role played by autophagy in the intracellular survival of B. pseudomallei in phagocytic and non-phagocytic cell lines. Autophagy was induced in response to B. pseudomallei invasion of murine macrophage (RAW 264.7) cells and a proportion of the bacteria co-localized with the autophagy effector protein LC3, a marker for autophagosome formation. Pharmacological stimulation of autophagy in RAW 264.7 and murine embryonic fibroblast (MEF) cell lines resulted in increased co-localization of B. pseudomallei with LC3 while basal levels of co-localization could be abrogated using inhibitors of the autophagic pathway. Furthermore, induction of autophagy decreased the intracellular survival of B. pseudomallei in these cell lines, but bacterial survival was not affected in MEF cell lines deficient in autophagy. Treatment of infected macrophages with chloramphenicol increased the proportion of bacteria within autophagosomes indicating that autophagic evasion is an active process relying on bacterial protein synthesis. Consistent with this hypothesis, we identified a B. pseudomallei type III secreted protein, BopA, which plays a role in mediating bacterial evasion of autophagy. We conclude that the autophagic pathway is a component of the innate defense system against invading B. pseudomallei, but which the bacteria can actively evade. However, when autophagy is pharmacologically induced using rapamycin, bacteria are actively sequestered in autophagosomes, ultimately decreasing their survival.
Asunto(s)
Autofagia/fisiología , Burkholderia pseudomallei/fisiología , Línea Celular/microbiología , Supervivencia Celular , Androstadienos/farmacología , Animales , Antibacterianos/farmacología , Autofagia/efectos de los fármacos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Biomarcadores/metabolismo , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/patogenicidad , Humanos , Ratones , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Sirolimus/farmacología , WortmaninaAsunto(s)
Burkholderia pseudomallei , Melioidosis , Animales , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Bioterrorismo , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/genética , Burkholderia pseudomallei/patogenicidad , Burkholderia pseudomallei/fisiología , Ensayos Clínicos como Asunto , Enfermedades Endémicas , Humanos , Melioidosis/diagnóstico , Melioidosis/tratamiento farmacológico , Melioidosis/epidemiología , Melioidosis/microbiología , Ratones , Prevalencia , Tailandia/epidemiología , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/fisiologíaRESUMEN
Melioidosis is a notoriously protracted illness and is difficult to cure. We hypothesize that the causative organism, Burkholderia pseudomallei, undergoes a process of adaptation involving altered expression of surface determinants which facilitates persistence in vivo and that this is reflected by changes in colony morphology. A colony morphotyping scheme and typing algorithm were developed using clinical B. pseudomallei isolates. Morphotypes were divided into seven types (denoted I to VII). Type I gave rise to other morphotypes (most commonly type II or III) by a process of switching in response to environmental stress, including starvation, iron limitation, and growth at 42 degrees C. Switching was associated with complex shifts in phenotype, one of which (type I to type II) was associated with a marked increase in production of factors putatively associated with in vivo concealment. Isogenic types II and III, derived from type I, were examined using several experimental models. Switching between isogenic morphotypes occurred in a mouse model, where type II appeared to become adapted for persistence in a low-virulence state. Isogenic type II demonstrated a significant increase in intracellular replication fitness compared with parental type I after uptake by epithelial cells in vitro. Isogenic type III demonstrated a higher replication fitness following uptake by macrophages in vitro, which was associated with a switch to type II. Mixed B. pseudomallei morphologies were common in individual clinical specimens and were significantly more frequent in samples of blood, pus, and respiratory secretions than in urine and surface swabs. These findings have major implications for therapeutics and vaccine development.
Asunto(s)
Burkholderia pseudomallei/genética , Burkholderia pseudomallei/patogenicidad , Animales , Adhesión Bacteriana/genética , Adhesión Bacteriana/fisiología , Burkholderia pseudomallei/citología , Células Epiteliales/microbiología , Genotipo , Humanos , Macrófagos/microbiología , Melioidosis/microbiología , Melioidosis/mortalidad , Ratones , Fenotipo , Tasa de Supervivencia , Virulencia/genéticaRESUMEN
Based on the morphological, physiological, biochemical and nutritional characteristics, cellular fatty acid and lipid composition, ubiquinone-8 as the major respiratory quinone, guanine-plus-cytosine content of DNA, DNA-DNA homology value, and sequence alignment of 16S rRNA nucleotides, Oklahoma isolate was reidentified as a strain of Pseudomonas pseudomallei.
Asunto(s)
Burkholderia pseudomallei/clasificación , Melioidosis/microbiología , Antibacterianos/farmacología , Composición de Base , Burkholderia pseudomallei/química , Burkholderia pseudomallei/citología , Burkholderia pseudomallei/fisiología , Metabolismo de los Hidratos de Carbono , ADN Bacteriano/química , Ácidos Grasos/análisis , Humanos , Lípidos/análisis , Masculino , Oklahoma , Fenotipo , Ubiquinona/análisisRESUMEN
A study was made on the growth and survival of Pseudomonas pseudomallei in culture environments differing in nutrients, initial pH, and aeration, in comparison with Pseudomonas cepacia and Pseudomonas aeruginosa. The observations led us to a view that P. pseudomallei has the highest adaptability to acidic environments among the three species. Unlike the other species, it grew in heart infusion broth of initial pH 4.5 under aeration and survived keeping a high level (10(9) per ml) of viable counts for as long as 30 days. This sort of adaptation was found to be more evident in the media of poor nutrition and under limited aeration.