Biofilm formed by a hypervirulent (hypermucoviscous) variant of Klebsiella pneumoniae does not enhance serum resistance or survival in an in vivo abscess model

• Published in: Virulence 2012-05, Vol.3 (3), p.309-318
• Main Authors: Kong, Qingli, Beanan, Janet M., Olson, Ruth, MacDonald, Ulrike, Shon, Alyssa S., Metzger, Daniel J., Pomakov, Alexander O., Russo, Thomas A.
• Format: Article
• Language: English
• Subjects: Abscess - microbiology; abscess model; animal model; Animals; Binding; biofilm; Biofilms - growth & development; Biology; Bioscience; Blood Bactericidal Activity; Calcium; Cancer; Cell; complement; Cycle; Disease Models, Animal; DNA Transposable Elements; Gene Knockout Techniques; Humans; hypermucoviscous; hypervirulent; Klebsiella Infections - microbiology; Klebsiella pneumoniae; Klebsiella pneumoniae - genetics; Klebsiella pneumoniae - immunology; Klebsiella pneumoniae - pathogenicity; Klebsiella pneumoniae - physiology; Landes; Male; Microbial Viability; Mutagenesis, Insertional; Organogenesis; Polysaccharides, Bacterial - metabolism; Proteins; Rats; Rats, Long-Evans; Research Paper; Serum - immunology; virulence; Virulence Factors - genetics; Virulence Factors - metabolism; Young Adult
• ISSN: 2150-5594; 2150-5608
• DOI: 10.4161/viru.20383

A new hypervirulent (hypermucoviscous) clinical variant of Klebsiella pneumoniae (hvKP) has emerged over the last decade. Our goal is to identify new mechanisms, which increase the virulence hvKP. It has been shown that hvKP strains produce more biofilm than "classical" stains of K. pneumoniae, therefore we hypothesized that biofilm formation may contribute to the pathogenesis of systemic infection. To test this hypothesis, transposon mutants of the model pathogen hvKP1 were generated and screened for decreased production of biofilm. Three mutant constructs with disruptions in glnA [putatively encodes glutamine synthetase, hvKP1 glnA:: EZ::TN < KAN-2 > (glnA::Tn)], sucD [putatively encodes succinyl-CoA synthase α subunit, hvKP1 sucD:: EZ::TN < KAN-2 > (sucD::Tn)], and tag [putatively encodes transcriptional antiterminator of glycerol uptake operon, hvKP1 tag:: EZ::TN < KAN-2 > (tag::Tn)] were chosen for further characterization and use in biologic studies. Quantitative assays performed in rich laboratory medium and human ascites confirmed the phenotype and a hypermucoviscosity assay established that capsule production was not affected. However, compared with its wild-type parent, neither planktonic cells nor biofilms of glnA::Tn, sucD::Tn and tag::Tn displayed a change to the bactericidal activity of 90% human serum. Likewise, when assessed in a rat subcutaneous abscess model, the growth and survival of glnA::Tn, sucD::Tn and tag::Tn in abscess fluid was similar to hvKP1. In this report we identify three new genes that contribute to biofilm formation in hvKP1. However, decreased biofilm production due to disruption of these genes does not affect the sensitivity of these mutant constructs to 90% human serum when in planktonic form or within a biofilm. Further, their virulence in an in vivo abscess model was unaffected.