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Crenarchaeal biofilm formation under extreme conditions.

Koerdt A, Gödeke J, Berger J, Thormann KM, Albers SV - PLoS ONE (2010)

Bottom Line: However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments.While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells.The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biology of Archaea, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.

ABSTRACT

Background: Biofilm formation has been studied in much detail for a variety of bacterial species, as it plays a major role in the pathogenicity of bacteria. However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments.

Methodology: We have analyzed biofilm formation in three closely related hyperthermophilic crenarchaeotes: Sulfolobus acidocaldarius, S. solfataricus and S. tokodaii. We established a microtitre plate assay adapted to high temperatures to determine how pH and temperature influence biofilm formation in these organisms. Biofilm analysis by confocal laser scanning microscopy demonstrated that the three strains form very different communities ranging from simple carpet-like structures in S. solfataricus to high density tower-like structures in S. acidocaldarius in static systems. Lectin staining indicated that all three strains produced extracellular polysaccharides containing glucose, galactose, mannose and N-acetylglucosamine once biofilm formation was initiated. While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells.

Conclusion: The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

Show MeSH
Different structures of static biofilms formed by three Sulfolobus strains S. acidocaldarius, S. solfataricus and S. tokodaii visualized by CLSM and SEM.A (top views) and B (side views) display the overlays of the images of three day old biofilms treated with DAPI and DDAO. The bar is 20 µm in length. C (top view) and D (side views) show the single channels of the overlays. DAPI signal: blue; DDAO signal: yellow. E, SEM images of biofilms of the three Sulfolobus strains incubated for 6 days. CLSM: confocal laser scanning microscopy; SEM: scanning electron microscopy.
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pone-0014104-g002: Different structures of static biofilms formed by three Sulfolobus strains S. acidocaldarius, S. solfataricus and S. tokodaii visualized by CLSM and SEM.A (top views) and B (side views) display the overlays of the images of three day old biofilms treated with DAPI and DDAO. The bar is 20 µm in length. C (top view) and D (side views) show the single channels of the overlays. DAPI signal: blue; DDAO signal: yellow. E, SEM images of biofilms of the three Sulfolobus strains incubated for 6 days. CLSM: confocal laser scanning microscopy; SEM: scanning electron microscopy.

Mentions: All three Sulfolobus strains were inoculated in uncoated plastic µ-dishes and incubated without agitation at 76°C. Evaporation was prevented by placing the Petri dishes in a humidified metal box and the medium was carefully exchanged every 24 hrs with fresh, prewarmed medium to ensure nutrient and oxygen availability. After three days the formed biofilms were stained with DAPI, as described in the Materials and Methods section and analyzed by confocal laser scanning microscopy (CLSM). We employed DAPI staining to visualize cells as there is currently no fluorescent protein available that is stably expressed under the growth conditions of Sulfolobus spp.. S. solfataricus formed biofilms (20–30 µm thick) with a carpet like structure covering the whole surface of the Petri dish with a low density of cells (Fig. 2, middle column). The biofilm structure of S. tokodaii was 25–35 µm thick and also exhibited a carpet like structure but, in contrast to S. solfataricus, these had a high cell density and, occasionally, cell aggregates were visible (Fig. 3, overlay picture, last row). S. acidocaldarius readily formed biofilms (25–35 µm thick) which contained a high density of cells and large aggregates, forming towering structures above the surface of attached cells (Fig. 3, first row).


Crenarchaeal biofilm formation under extreme conditions.

Koerdt A, Gödeke J, Berger J, Thormann KM, Albers SV - PLoS ONE (2010)

Different structures of static biofilms formed by three Sulfolobus strains S. acidocaldarius, S. solfataricus and S. tokodaii visualized by CLSM and SEM.A (top views) and B (side views) display the overlays of the images of three day old biofilms treated with DAPI and DDAO. The bar is 20 µm in length. C (top view) and D (side views) show the single channels of the overlays. DAPI signal: blue; DDAO signal: yellow. E, SEM images of biofilms of the three Sulfolobus strains incubated for 6 days. CLSM: confocal laser scanning microscopy; SEM: scanning electron microscopy.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2991349&req=5

pone-0014104-g002: Different structures of static biofilms formed by three Sulfolobus strains S. acidocaldarius, S. solfataricus and S. tokodaii visualized by CLSM and SEM.A (top views) and B (side views) display the overlays of the images of three day old biofilms treated with DAPI and DDAO. The bar is 20 µm in length. C (top view) and D (side views) show the single channels of the overlays. DAPI signal: blue; DDAO signal: yellow. E, SEM images of biofilms of the three Sulfolobus strains incubated for 6 days. CLSM: confocal laser scanning microscopy; SEM: scanning electron microscopy.
Mentions: All three Sulfolobus strains were inoculated in uncoated plastic µ-dishes and incubated without agitation at 76°C. Evaporation was prevented by placing the Petri dishes in a humidified metal box and the medium was carefully exchanged every 24 hrs with fresh, prewarmed medium to ensure nutrient and oxygen availability. After three days the formed biofilms were stained with DAPI, as described in the Materials and Methods section and analyzed by confocal laser scanning microscopy (CLSM). We employed DAPI staining to visualize cells as there is currently no fluorescent protein available that is stably expressed under the growth conditions of Sulfolobus spp.. S. solfataricus formed biofilms (20–30 µm thick) with a carpet like structure covering the whole surface of the Petri dish with a low density of cells (Fig. 2, middle column). The biofilm structure of S. tokodaii was 25–35 µm thick and also exhibited a carpet like structure but, in contrast to S. solfataricus, these had a high cell density and, occasionally, cell aggregates were visible (Fig. 3, overlay picture, last row). S. acidocaldarius readily formed biofilms (25–35 µm thick) which contained a high density of cells and large aggregates, forming towering structures above the surface of attached cells (Fig. 3, first row).

Bottom Line: However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments.While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells.The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biology of Archaea, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.

ABSTRACT

Background: Biofilm formation has been studied in much detail for a variety of bacterial species, as it plays a major role in the pathogenicity of bacteria. However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments.

Methodology: We have analyzed biofilm formation in three closely related hyperthermophilic crenarchaeotes: Sulfolobus acidocaldarius, S. solfataricus and S. tokodaii. We established a microtitre plate assay adapted to high temperatures to determine how pH and temperature influence biofilm formation in these organisms. Biofilm analysis by confocal laser scanning microscopy demonstrated that the three strains form very different communities ranging from simple carpet-like structures in S. solfataricus to high density tower-like structures in S. acidocaldarius in static systems. Lectin staining indicated that all three strains produced extracellular polysaccharides containing glucose, galactose, mannose and N-acetylglucosamine once biofilm formation was initiated. While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells.

Conclusion: The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

Show MeSH