Examination of reproducibility in microbiological degradation experiments

Experimental data indicate that certain microbiological degradation experiments have a limited reproducibility. Nine identical batch experiments were carried out on 3 different days to examine reproducibility. A pure culture, isolated from soil, grew with toluene as the only carbon and energy source. Toluene was degraded under aerobic conditions at a constant temperature of 28 degrees C. The experiments were modelled by a Monod model--extended to meet the air/liquid system, and the parameter values were estimated using a statistical nonlinear estimation procedure. Model reduction analysis resulted in a simpler model without the biomass decay term. In order to test for model reduction and reproducibility of parameter estimates, a likelihood ratio test was employed. The limited reproducibility for these experiments implied that all 9 batch experiments could not be described by the same set of parameter values. However, experiments carried out the same day (within the same run) were more uniform than experiments carried out on different days (between runs), and a common set of parameter estimates could be accepted for experiments within runs, but not for experiments from different runs. The limited reproducibility may be caused by variability in the preculture, or more precisely, variations in the physiological state of the bacteria in the precultures just before used as inoculum.