Two stable variants of Burkholderia pseudomallei strain MSHR5848 express broadly divergent in vitro phenotypes associated with their virulence differences

Autor: Robert C. Bernhards, Jeffrey W. Koehler, Sean Lovett, Ronald G. Toothman, Christopher P. Klimko, Joel A. Bozue, Jason T. Ladner, Susan L. Welkos, Steven J. Kern, David P Fetterer, G. I. Koroleva, Gustavo Palacios, Patricia L. Worsham, Mark Wolcott, David A. Rozak, C. J. Chase, Christopher K. Cote, April A. Shea
Rok vydání: 2017
Předmět:
Bacterial Diseases
0301 basic medicine
Burkholderia pseudomallei
lcsh:Medicine
Pathology and Laboratory Medicine
Mice
White Blood Cells
Ribotyping
Animal Cells
Antibiotics
Medicine and Health Sciences
lcsh:Science
Staining
Mice
Inbred BALB C
Multidisciplinary
Virulence
biology
Antimicrobials
Drugs
Phenotype
Bacterial Pathogens
Membrane Staining
Phenotypes
Chemistry
Infectious Diseases
Medical Microbiology
Physical Sciences
Cellular Types
Pathogens
Research Article
Chemical Elements
Virulence Factors
Burkholderia
Immune Cells
Immunology
030106 microbiology
Research and Analysis Methods
Microbiology
Cell Line
03 medical and health sciences
Microbial Control
Drug Resistance
Bacterial
Genetics
Animals
Microbial Pathogens
Gene
Pharmacology
Polymorphism
Genetic
Blood Cells
Bacteria
Macrophages
lcsh:R
Organisms
Correction
Biology and Life Sciences
Phenotype microarray
Cell Biology
biology.organism_classification
030104 developmental biology
Genes
Bacterial
Specimen Preparation and Treatment
Burkholderia Infection
Multilocus sequence typing
lcsh:Q
Gram Staining
Sulfur
Zdroj: PLoS ONE
PLoS ONE, Vol 12, Iss 2, p e0171363 (2017)
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0171363
Popis: Burkholderia pseudomallei (Bp), the agent of melioidosis, causes disease ranging from acute and rapidly fatal to protracted and chronic. Bp is highly infectious by aerosol, can cause severe disease with nonspecific symptoms, and is naturally resistant to multiple antibiotics. However, no vaccine exists. Unlike many Bp strains, which exhibit random variability in traits such as colony morphology, Bp strain MSHR5848 exhibited two distinct and relatively stable colony morphologies on sheep blood agar plates: a smooth, glossy, pale yellow colony and a flat, rough, white colony. Passage of the two variants, designated "Smooth" and "Rough", under standard laboratory conditions produced cultures composed of > 99.9% of the single corresponding type; however, both could switch to the other type at different frequencies when incubated in certain nutritionally stringent or stressful growth conditions. These MSHR5848 derivatives were extensively characterized to identify variant-associated differences. Microscopic and colony morphology differences on six differential media were observed and only the Rough variant metabolized sugars in selective agar. Antimicrobial susceptibilities and lipopolysaccharide (LPS) features were characterized and phenotype microarray profiles revealed distinct metabolic and susceptibility disparities between the variants. Results using the phenotype microarray system narrowed the 1,920 substrates to a subset which differentiated the two variants. Smooth grew more rapidly in vitro than Rough, yet the latter exhibited a nearly 10-fold lower lethal dose for mice than Smooth. Finally, the Smooth variant was phagocytosed and replicated to a greater extent and was more cytotoxic than Rough in macrophages. In contrast, multiple locus sequence type (MLST) analysis, ribotyping, and whole genome sequence analysis demonstrated the variants' genetic conservation; only a single consistent genetic difference between the two was identified for further study. These distinct differences shown by two variants of a Bp strain will be leveraged to better understand the mechanism of Bp phenotypic variability and to possibly identify in vitro markers of infection.
Databáze: OpenAIRE
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