Bacteria that exist in a biofilm are phenotypically different from suspended cells of the same genotype. The study of biofilm in the laboratory requires protocols that account for this difference. Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. The purpose of this method is to direct a user in the laboratory study of biofilms by clearly defining each system parameter. This method will enable a person to grow, sample, and analyze a laboratory biofilm.

1.1 This test method is used for growing a reproducible (1) Pseudomonas aeruginosa biofilm in a continuously stirred tank reactor (CSTR) under medium shear conditions. In addition, the test method describes how to sample and analyze biofilm for viable cells.

1.2 Although this test method was created to mimic conditions within a toilet bowl, it can be adapted for the growth and characterization of varying species of biofilm (rotating disk reactorrepeatability and relevance (2)).

1.3 This test method describes how to sample and analyze biofilm for viable cells. Biofilm population density is recorded as log₁₀ colony forming units per surface area (rotating disk reactorefficacy test method (3)).

1.4 Basic microbiology training is required to perform this test method.

1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.

1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.