Microtiter susceptibility testing of microbes growing on peg lids: a miniaturized biofilm model for high-throughput screening.
Batch culture of biofilms on peg lids is a versatile method that can be used for microtiter determinations of biofilm antimicrobial susceptibility. In this paper, we describe a core protocol and a set of parameters (surface composition, the rate of rocking or orbital motion, temperature, cultivation time, inoculum size, atmospheric gases and nutritional medium) that can be adjusted to grow single- or multispecies biofilms on peg surfaces. Mature biofilms formed on peg lids can then be fitted into microtiter plates containing test agents. After a suitable exposure time, biofilm cells are disrupted into a recovery medium using sonication. Microbicidal endpoints can be determined qualitatively using optical density measurements or quantitatively using viable cell counting. Once equipment is calibrated and growth conditions are at an optimum, the procedure requires approximately 5 h of work over 4-6 d. This efficient method allows antimicrobial agents and exposure conditions to be tested against biofilms on a high-throughput scale.
Harrison
Joe J
JJ
Department of Microbiology, University of Washington School of Medicine, Seattle, Washington, USA. jjharris@u.washington.edu
Stremick
Carol A
CA
Turner
Raymond J
RJ
Allan
Nick D
ND
Olson
Merle E
ME
Ceri
Howard
H
D000890:Anti-Infective Agents / Q000494:pharmacology; D001422:Bacterial Adhesion; D001431:Bacteriological Techniques / Q000379:methods*; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development*; D057166:High-Throughput Screening Assays / Q000379:methods*; D008087:Listeria / Q000187:drug effects / Q000254:growth & development; D008826:Microbial Sensitivity Tests / Q000379:methods*; D013499:Surface Properties; D013696:Temperature
D000890:Anti-Infective Agents