Macrophage and Galleria mellonella infection models reflect the virulence of naturally occurring isolates of B. pseudomallei, B. thailandensis and B. oklahomensis

BackgroundBurkholderia pseudomallei is the causative agent of melioidosis, a tropical disease of humans with a variable and often fatal outcome. In murine models of infection, different strains exhibit varying degrees of virulence. In contrast, two related species, B. thailandensis and B. oklahomensis, are highly attenuated in mice. Our aim was to determine whether virulence in mice is reflected in macrophage or wax moth larvae (Galleria mellonella) infection models.ResultsB. pseudomallei strains 576 and K96243, which have low median lethal dose (MLD) values in mice, were able to replicate and induce cellular damage in macrophages and caused rapid death of G. mellonella. In contrast, B. pseudomallei strain 708a, which is attenuated in mice, showed reduced replication in macrophages, negligible cellular damage and was avirulent in G. mellonella larvae. B. thailandensis isolates were less virulent than B. pseudomallei in all of the models tested. However, we did record strain dependent differences. B. oklahomensis isolates were the least virulent isolates. They showed minimal ability to replicate in macrophages, were unable to evoke actin-based motility or to form multinucleated giant cells and were markedly attenuated in G. mellonella compared to B. thailandensis.ConclusionsWe have shown that the alternative infection models tested here, namely macrophages and Galleria mellonella, are able to distinguish between strains of B. pseudomallei, B. thailandensis and B. oklahomensis and that these differences reflect the observed virulence in murine infection models. Our results indicate that B. oklahomensis is the least pathogenic of the species investigated. They also show a correlation between isolates of B. thailandensis associated with human infection and virulence in macrophage and Galleria infection models.

[1]  W. Nierman,et al.  Bacterial genome adaptation to niches: Divergence of the potential virulence genes in three Burkholderia species of different survival strategies , 2005, BMC Genomics.

[2]  Joanne M Stevens,et al.  Identification of a bacterial factor required for actin‐based motility of Burkholderia pseudomallei , 2005, Molecular microbiology.

[3]  M. Schell,et al.  Comparative Genomics and an Insect Model Rapidly Identify Novel Virulence Genes of Burkholderia mallei , 2008, Journal of bacteriology.

[4]  S. Sirisinha,et al.  Virulent Burkholderia pseudomallei is more efficient than avirulent Burkholderia thailandensis in invasion of and adherence to cultured human epithelial cells. , 2004, Microbial pathogenesis.

[5]  J. Badger,et al.  Continuing Evolution of Burkholderia mallei Through Genome Reduction and Large-Scale Rearrangements , 2010, Genome biology and evolution.

[6]  P. Tan,et al.  Identification of a regulatory cascade controlling Type III Secretion System 3 gene expression in Burkholderia pseudomallei , 2010, Molecular microbiology.

[7]  S. Peacock,et al.  Management of melioidosis , 2006, Expert review of anti-infective therapy.

[8]  Yahua Chen,et al.  Identification of tomato plant as a novel host model for Burkholderia pseudomallei , 2010, BMC Microbiology.

[9]  A. Steigerwalt,et al.  Burkholderia oklahomensis sp. nov., a Burkholderia pseudomallei-like species formerly known as the Oklahoma strain of Pseudomonas pseudomallei. , 2006, International journal of systematic and evolutionary microbiology.

[10]  S. Cardona,et al.  An expression vector containing a rhamnose-inducible promoter provides tightly regulated gene expression in Burkholderia cenocepacia. , 2005, Plasmid.

[11]  D. DeShazer.  Virulence of clinical and environmental isolates of Burkholderia oklahomensis and Burkholderia thailandensis in hamsters and mice. , 2007, FEMS microbiology letters.

[12]  Nicholas J. White,et al.  Melioidosis: insights into the pathogenicity of Burkholderia pseudomallei , 2006, Nature Reviews Microbiology.

[13]  G. Dougan,et al.  Characterisation of an acapsular mutant of Burkholderia pseudomallei identified by signature tagged mutagenesis. , 2002, Journal of medical microbiology.

[14]  A. Steigerwalt,et al.  Pneumonia and Septicemia Caused by Burkholderia thailandensis in the United States , 2006, Journal of Clinical Microbiology.

[15]  J. Boyce,et al.  Wound infection by an indigenous Pseudomonas pseudomallei-like organism isolated from the soil: case report and epidemiologic study. , 1977, The Journal of infectious diseases.

[16]  David DeShazer,et al.  Genomic patterns of pathogen evolution revealed by comparison of Burkholderia pseudomallei, the causative agent of melioidosis, to avirulent Burkholderia thailandensis , 2006, BMC Microbiology.

[17]  D. DeShazer,et al.  Burkholderia thailandensis sp. nov., a Burkholderia pseudomallei-like species. , 1998, International journal of systematic bacteriology.

[18]  G. Crawford,et al.  Cutaneous Melioidosis in a Man Who Was Taken as a Prisoner of War by the Japanese during World War II , 2005, Journal of Clinical Microbiology.

[19]  Samuel I. Miller,et al.  Burkholderia thailandensis as a Model System for the Study of the Virulence-Associated Type III Secretion System of Burkholderia pseudomallei , 2008, Infection and Immunity.

[20]  Joanne M Stevens,et al.  Actin-Binding Proteins from Burkholderia mallei and Burkholderia thailandensis Can Functionally Compensate for the Actin-Based Motility Defect of a Burkholderia pseudomallei bimA Mutant , 2005, Journal of bacteriology.

[21]  W. Wiersinga,et al.  MELIOIDOSIS , 1975, The Lancet.

[22]  Wing-Kin Sung,et al.  Genomic acquisition of a capsular polysaccharide virulence cluster by non-pathogenic Burkholderia isolates , 2010, Genome Biology.

[23]  M. Stevens,et al.  An Inv/Mxi‐Spa‐like type III protein secretion system in Burkholderia pseudomallei modulates intracellular behaviour of the pathogen , 2002, Molecular microbiology.

[24]  B. Currie,et al.  Animal melioidosis in Australia. , 2000, Acta tropica.

[25]  D. DeShazer,et al.  Detection of Bacterial Virulence Genes by Subtractive Hybridization: Identification of Capsular Polysaccharide ofBurkholderia pseudomallei as a Major Virulence Determinant , 2001, Infection and Immunity.

[26]  R. Finkelstein,et al.  Pseudomonas (Burkholderia) pseudomallei in Thailand, 1964-1967: geographic distribution of the organism, attempts to identify cases of active infection, and presence of antibody in representative sera. , 2000, The American journal of tropical medicine and hygiene.

[27]  A. Engering,et al.  Differential intracellular fate of Burkholderia pseudomallei 844 and Burkholderia thailandensis UE5 in human monocyte-derived dendritic cells and macrophages , 2009, BMC Immunology.

[28]  R. Kinoshita,et al.  Pathology of melioidosis in captive marine mammals. , 2000, Australian veterinary journal.

[29]  Kim Rutherford,et al.  Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[30]  J. Jeddeloh,et al.  Burkholderia pseudomallei kills the nematode Caenorhabditis elegans using an endotoxin‐mediated paralysis , 2001, Cellular microbiology.

[31]  R. Roop,et al.  pBBR1MCS: a broad-host-range cloning vector. , 1994, BioTechniques.

[32]  S. Beckstrom-Sternberg,et al.  Molecular Basis of Rare Aminoglycoside Susceptibility and Pathogenesis of Burkholderia pseudomallei Clinical Isolates from Thailand , 2009, PLoS neglected tropical diseases.

[33]  Richard A. Moore,et al.  Genome-wide expression analysis of iron regulation in Burkholderia pseudomallei and Burkholderia mallei using DNA microarrays. , 2005, FEMS microbiology letters.

[34]  J. H. Crosa,et al.  Genetic and Transcriptional Analysis of the Siderophore Malleobactin Biosynthesis and Transport Genes in the Human Pathogen Burkholderia pseudomallei K96243 , 2006, Journal of bacteriology.

[35]  A. Haque,et al.  Burkholderia pseudomallei: animal models of infection. , 2008, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[36]  B. Drasar,et al.  Effects of Burkholderia pseudomallei and other Burkholderia species on eukaryotic cells in tissue culture. , 1998, Microbios.

[37]  A. Jones,et al.  Intracellular survival of Burkholderia pseudomallei , 1996, Infection and immunity.

[38]  S. Sirisinha,et al.  Burkholderia pseudomallei Induces Cell Fusion and Actin-Associated Membrane Protrusion: a Possible Mechanism for Cell-to-Cell Spreading , 2000, Infection and Immunity.

[39]  Julian J. Nussbaum,et al.  Pseudomonas pseudomallel in an Anophthalmic Orbit , 1980 .

[40]  R. Ulrich,et al.  The Animal Pathogen-Like Type III Secretion System Is Required for the Intracellular Survival of Burkholderia mallei within J774.2 Macrophages , 2006, Infection and Immunity.