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Francisella novicida Forms In Vitro Biofilms Mediated by an Orphan Response Regulator

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Abstract

Francisella tularensis is associated with water and waterways and infects many species of animals, insects, and protists. The mechanism Francisella utilizes to persist in the environment and in tick vectors is currently unknown. We have demonstrated for the first time that Francisella novicida, a model organism of F. tularensis, forms a biofilm in vitro. Selected F. novicida transposon mutants were tested for their ability to form biofilm compared to the wildtype F. novicida strain. Mutation of the putative qseB gene led to an impairment in the ability to form biofilm with no impairment in bacterial growth. A qseC mutant had impaired growth but demonstrated a marked impairment in biofilm production. Mutation in capC affected both bacterial growth and biofilm formation, but no biofilm production impairment was seen with capB or pilE mutants. A deletion mutant in the orphan response regulator FTN_1465, which we propose is the putative QseB, formed significantly less biofilm than the wildtype. When FTN_1465 was complemented back into the deletion mutant, biofilm formation was restored. Thus, the orphan response regulator FTN_1465 is an important factor in biofilm production in vitro in F. novicida. These results demonstrate that Francisella species are able to form biofilms in vitro, suggesting that biofilm formation may be important for the lifecycle of this organism.

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Abbreviations

EPM:

Extracellular polysaccharide matrix

TSB-C:

Tryptic soy broth supplemented with 0.1% cysteine HCl

CFU:

Colony forming units

LPS:

Lipopolysaccharide

Pf:

Pseudomonas fluorescens

Fn:

Francisella novicida

REC:

Signal receiver domain

PS:

Polystyrene

PVC:

Polyvinylchloride

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Acknowledgements

MDC was supported by a Graduate Research Assistantship from the National Center for Biodefense and Infectious Diseases, George Mason University, and the National Science Foundation GK-12 Fellowship. ABV was supported by a High-Potential Graduate Research Assistantship from the Office of the Provost, George Mason University, and was partially supported through the Joint Science and Technology Office for Chemical and Biological Defense/Defense Threat Reduction Agency and contracted through the US Army Medical Research and Material Command under Contract No. W81XWH-06-C-0306. This research was also supported by a Small Research Grant to MVH from the Virginia Academy of Science. The confocal microscope was purchased with support from DOE Grant DE-F C52-04NA25455 and the Commonwealth of Virginia Educational Trust Fund. P. fluorescens (Pf) was a kind gift from J. Mose and Dr. P. Royt (GMU). ΔFTN_1465 and the ΔFTN_1465 complement were a kind gift from Dr. J. S. Gunn (The Ohio State University Medical Center). We would like to thank Dr. D. Cox (GMU) for technical assistance with the confocal microscope.

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  1. MS 1HS Department of Molecular and Microbiology, National Center for Biodefense and Infectious Diseases, George Mason University, 10900 University Blvd, Manassas, VA, 20110, USA

    Meghan W. Durham-Colleran, Anne Brooks Verhoeven & Monique L. van Hoek

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  1. Meghan W. Durham-Colleran
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Correspondence to Monique L. van Hoek.

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Durham-Colleran, M.W., Verhoeven, A.B. & van Hoek, M.L. Francisella novicida Forms In Vitro Biofilms Mediated by an Orphan Response Regulator. Microb Ecol 59, 457–465 (2010). https://doi.org/10.1007/s00248-009-9586-9

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