Effect of nutrient limitation on adhesion characteristics of Pseudomonas aeruginosa.
Pseudomonas aeruginosa causes a variety of diseases in humans including lung and ocular infections. Infections of the cornea are usually associated with wearing contact lenses and can result in loss of vision. This study aimed to determine the effect of carbon or nitrogen limitation on the adhesion to contact lenses of a strain of Ps. aeruginosa isolated from contact lens-related corneal inflammation. Cells were grown in a continuous culture apparatus in varying levels of glucose or ammonia to effect nutrient limitation. Adhesion to contact lenses was measured as total counts and viable counts. The cell surface hydrophobicity and charge were measured using adhesion to surface-modified Sepharose. Changes in lipopolysaccharide were determined using 1D SDS-PAGE and changes in cell-surface proteins were measured using 2D gel electrophoresis. The more the cultures were nitrogen limited, the greater the increase in adhesion to unworn hydrogel contact lenses 0.3 x 10(3) - 2.2 x 10(3) cells/mm2 on Etafilon A lenses. Cells that were carbon limited showed a greater increase in adhesion to contact lenses when the lenses had been coated in artificial tears. It appeared that lipopolysaccharide may have been involved in the constitutive adhesion to unworn lenses that occurred during C-limitation, whereas changes in the outer membrane proteins contributed to the increased adhesion under nitrogen limitation, or the change in adhesion that occurred to carbon-limited cells using contact lenses coated in artificial tears. Nine cell-surface proteins appeared during nitrogen limitation with kDa/pI of 75/4.8, 4.9, 5.0; 62/5.6; 89/6.5; 38/6.4; 28/1.5; 18/6.4; 12/4.5. Any or all of these may have been involved in the increased adhesion and further experiments are underway to examine this possibility.
Cowell
B A
BA
Cooperative Research Centre for Eye Research and Technology, University of New South Wales, Australia.
Willcox
M D
MD
Herbert
B
B
Schneider
R P
RP
D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D003261:Contact Lenses / Q000382:microbiology*; D003470:Culture Media; D015180:Electrophoresis, Gel, Two-Dimensional; D015818:Eye Infections, Bacterial / Q000382:microbiology; D006801:Humans; D020100:Hydrogels; D008070:Lipopolysaccharides / Q000302:isolation & purification / Q000378:metabolism; D008854:Microscopy, Electron; D009584:Nitrogen / Q000378:metabolism; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000502:physiology*
D001425:Bacterial Outer Membrane Proteins; D003470:Culture Media; D020100:Hydrogels; D008070:Lipopolysaccharides; D002244:Carbon; D009584:Nitrogen