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Chapter 93 : Biofilm Formation of in Complex Medium under Static and Dynamic Flow Conditions

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Biofilm Formation of in Complex Medium under Static and Dynamic Flow Conditions, Page 1 of 2

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Abstract:

This chapter analyzes biofilm formation of in rich medium which supports extracellular proliferation of the bacteria. To investigate biofilm formation of under static conditions, a 1:1 mixture of labeled with the fluorescent proteins EGFP or DsRed-Express was inoculated in a glass-bottom 35-mm petri dish. Biofilm formation was analyzed with an inverted confocal microscope (Axiovert 200M, 100 X oil objective Plan Neofluar; Zeiss). Biofilm formation of in rich medium was also quantified by crystal violet staining in upright polystyrene microtiter plates or on polystyrene pins of "inverse" lids under static (no medium exchange) or quasi-static conditions (medium replaced twice a day). The mutant strain reproducibly accumulated 30% less biomass within 5 days, demonstrating that bacterial factors contribute to biofilm formation. It is noteworthy that mutants lacking or , both of which are required for the expression of transmissive (virulence) traits of , were not affected in biofilm formation. A continuous-flow chamber system was set up to further test the hypothesis that plank-tonic cells are crucial for biofilm formation on surfaces and to noninvasively monitor in real-time the adherence and biofilm formation of under dynamic flow conditions.

Citation: Mampel J, Spirig T, S. Weber S, A. J. Haagensen J, Molin S, Hilbi H. 2006. Biofilm Formation of in Complex Medium under Static and Dynamic Flow Conditions, p 398-402. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch93

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Figures

Image of FIGURE 1
FIGURE 1

Biofilm formation and distribution of EGFP- and DsRedExpress-labeled under static conditions in rich medium. Confocal laser scanning micrograph of a representative biofilm section formed after 3 days in glass-bottom dishes by wild-type JR32 labeled with enhanced green fluorescent protein (EGFP, white) or with the red fluorescent protein DsRedExpress (black). Three-dimensional reconstruction of the image data was performed with the “Volocity” 2.6.1 software (Improvision). The strains were inoculated in a 1:1 mixture and remained homogeneously dispersed throughout the experiment without forming extended patches of clonally grown aggregates. Unit cell: 8.1 μm.

Citation: Mampel J, Spirig T, S. Weber S, A. J. Haagensen J, Molin S, Hilbi H. 2006. Biofilm Formation of in Complex Medium under Static and Dynamic Flow Conditions, p 398-402. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch93
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Image of FIGURE 2
FIGURE 2

Microcolonies formed by in a continuous flow chamber system in rich medium. The fluorescence micrographs show (A) compact microcolonies of GFP-labeled wild-type strain JR32 occasionally formed after 18 h and (B) filamentous microcolonies (“streamers”) present after 5 days. (C) Three-dimensional reconstruction of the filamentous microcolony shown in panel B was done as described in the legend to Fig. 1 . Bar (A, B), 100 μm. Unit cell (C), 23 μm.

Citation: Mampel J, Spirig T, S. Weber S, A. J. Haagensen J, Molin S, Hilbi H. 2006. Biofilm Formation of in Complex Medium under Static and Dynamic Flow Conditions, p 398-402. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch93
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References

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