title: Diversity of biofilms produced by quorum-sensing-deficient clinical isolates of Pseudomonas aeruginosa.
abstract: The quorum-sensing (QS) systems control several virulence attributes of Pseudomonas aeruginosa. Five QS-deficient P. aeruginosa clinical isolates (CI) that were obtained from wound (CI-1), tracheal (CI-2, CI-3, CI-4) and urinary tract (CI-5) infections had previously been characterized. In this study, a flow-through continuous-culture system was utilized to examine in detail the biofilms formed by these isolates in comparison with the P. aeruginosa prototrophic strain PAO1. Analysis of the biofilms by confocal laser scanning microscopy and COMSTAT image analysis at 1 and 7 days post-inoculation showed that the isolates produced diverse biofilms. In comparison with PAO1, the CI produced biofilms that scarcely or partially covered the surface at day 1, although CI-1 produced larger microcolonies. At day 7, CI-2 and CI-4 produced mature biofilms denser than that produced by PAO1, while the biofilm formed by CI-1 changed very little from day 1. CI-1 was defective in both swarming and twitching motilities, and immunoblotting analysis confirmed that it produced a reduced level of PilA protein. The twitching-motility defect of CI-1 was not complemented by a plasmid carrying intact pilA. In the 48 h colony biofilm assay, the CI varied in susceptibility to imipenem, gentamicin and piperacillin/tazobactam. These results suggest that: (1) the isolates produced biofilms with different structures and densities from that of PAO1; (2) biofilm formation by the isolates was not influenced by either the isolation site or the QS deficiencies of the isolates; (3) the behaviour of CI-1 in the different biofilm systems may be due to its lack of swarming motility and type IV pilus-related twitching motility.
authors.affiliation: Department of Surgery, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.|Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: J Andy
authors.initials: JA
authors.lastname: Schaber
authors.affiliation: Department of Surgery, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.|Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Adrienne
authors.initials: A
authors.lastname: Hammond
authors.affiliation: Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Nancy L
authors.initials: NL
authors.lastname: Carty
authors.affiliation: Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Simon C
authors.initials: SC
authors.lastname: Williams
authors.affiliation: Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Jane A
authors.initials: JA
authors.lastname: Colmer-Hamood
authors.affiliation: Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Ben H
authors.initials: BH
authors.lastname: Burrowes
authors.affiliation: School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Vijian
authors.initials: V
authors.lastname: Dhevan
authors.affiliation: Department of Surgery, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.|Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Griswold
authors.affiliation: Department of Surgery, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.|Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Abdul N
authors.initials: AN
authors.lastname: Hamood
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D018441:Biofilms / Q000254:growth & development*; D015169:Colony Count, Microbial; D005816:Genetic Complementation Test; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D008124:Locomotion / Q000235:genetics; D050296:Microbial Viability; D018613:Microscopy, Confocal; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development* / Q000502:physiology; D053038:Quorum Sensing / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins