Radiochemical assay to measure the biofilm produced by coagulase-negative staphylococci on solid surfaces and its use to quantitate the effects of various antibacterial compounds on the formation of the biofilm.
A firmly adherent mass of slime plus organisms (biofilm) accumulates on the sides of culture tubes when some strains of coagulase-negative staphylococci are grown in a chemically-defined medium containing [14C]glucose. This mass was washed (to remove labelled medium) and then counted after adding scintillation fluid. Organisms from the liquid culture were also washed and counted to check that [14C]glucose had been utilised to label the bacteria. Nine strains were examined in this way, and the results were compared with those obtained with four older techniques for recognising slime production or adherent bacteria. The new method is quick, and has advantages of reproducibility and good discrimination between strains; there was a 15-fold difference in counts in the biofilm between slime-producing and non-producing strains respectively. With the new radiolabel assay, the effects of several antibacterial compounds on the build-up of the biofilm were investigated with four slime-producing strains. Tunicamycin, chloramphenicol and 5-fluorouracil, at levels below their minimum growth-inhibitory concentrations, each greatly diminished biofilm formation; several other drugs had less effect.
Hussain
M
M
Department of Molecular Biology and Biotechnology, The University, Western Bank, Sheffield.
Collins
C
C
Hastings
J G
JG
White
P J
PJ
D000255:Adenosine Triphosphate; D000423:Alcian Blue / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001422:Bacterial Adhesion; D015169:Colony Count, Microbial; D003208:Concanavalin A / Q000378:metabolism; D003470:Culture Media; D006801:Humans; D011107:Polylysine / Q000378:metabolism; D012588:Scintillation Counting; D013053:Spectrophotometry; D013212:Staphylococcus epidermidis / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D003470:Culture Media; D003208:Concanavalin A; D011107:Polylysine; D000255:Adenosine Triphosphate; D000423:Alcian Blue