Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation.
Extracellular polymers can facilitate the non-specific attachment of bacteria to surfaces and hold together developing biofilms. This study was undertaken to qualitatively and quantitatively compare the architecture of biofilms produced by Pseudomonas aeruginosa strain PAO1 and its alginate-overproducing (mucA22) and alginate-defective (algD) variants in order to discern the role of alginate in biofilm formation. These strains, PAO1, Alg+ PAOmucA22 and Alg- PAOalgD, tagged with green fluorescent protein, were grown in a continuous flow cell system to characterize the developmental cycles of their biofilm formation using confocal laser scanning microscopy. Biofilm Image Processing (BIP) and Community Statistics (COMSTAT) software programs were used to provide quantitative measurements of the two-dimensional biofilm images. All three strains formed distinguishable biofilm architectures, indicating that the production of alginate is not critical for biofilm formation. Observation over a period of 5 days indicated a three-stage development pattern consisting of initiation, establishment and maturation. Furthermore, this study showed that phenotypically distinguishable biofilms can be quantitatively differentiated.
Stapper
Andres Plata
AP
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Narasimhan
Giri
G
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Ohman
Dennis E
DE
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Barakat
Johnny
J
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Hentzer
Morten
M
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Molin
Søren
S
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Kharazmi
Arsalan
A
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Høiby
Niels
N
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
Mathee
Kalai
K
Department of Biological Sciences1 and School of Computer Science2, Florida International University, Miami, FL 33199, USA 3Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA 4Section of Molecular Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark 5Department of Clinical Microbiology, University Hospital of Copenhagen, DK-2100 Copenhagen, Denmark.
D000464:Alginates; D018441:Biofilms / Q000254:growth & development*; D005798:Genes, Bacterial; D017930:Genes, Reporter; D020723:Glucuronic Acid / Q000096:biosynthesis* / Q000502:physiology*; D049452:Green Fluorescent Proteins; D006603:Hexuronic Acids; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D009154:Mutation; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis / Q000502:physiology; D011550:Pseudomonas aeruginosa / Q000378:metabolism* / Q000502:physiology; D013194:Staining and Labeling
D000464:Alginates; D006603:Hexuronic Acids; D008164:Luminescent Proteins; D011135:Polysaccharides, Bacterial; D049452:Green Fluorescent Proteins; D020723:Glucuronic Acid