Reproducibility of biofilm processes and the meaning of steady state in biofilm reactors.

The need for reproducing biofilm processes is undisputable - the quality of biofilm research depends on this reproducibility. However, as many biofilm researchers know, long-term biofilm processes are notoriously difficult to reproduce. To avoid problems related to biofilm reproducibility two strategies are used: (1) to study very young biofilms that have accumulated for a few hours to a few days only, and (2) to run biofilm experiments only once. The first approach trades reproducibility for relevance because natural biofilms are usually older, often much older than a few days. This approach can be applied to answer questions relevant to initial events of biofilm formation but not questions relevant to long-term biofilm accumulation. The second approach conceals the problem of biofilm reproducibility. To assure reproducibility of biofilm processes, we methodically followed a procedure for growing biofilms in terms of microbial makeup, media composition, temperature, surface preparation, etc. Despite all this effort the reproducibility of our results for long term growth is unimpressive. Consequently, the question had to be asked: Are biofilm processes reproducible? The experiments described in this paper address this question. Biofilms grown in two identical and identically operated biofilm reactors had comparable structure only until the first sloughing event. After that, biofilms had different patterns of accumulation.