As a representative member of the toluene-degrading population in a biofilter for waste gas treatment, Pseudomonas putida was investigated with a 16S rRNA targeting probe. The three-dimensional distribution of P. putida was visualized in the biofilm matrix by scanning confocal laser microscopy, demonstrating that P. putida was present throughout the biofilm. Acridine orange staining revealed a very heterogeneous structure of the fully hydrated biofilm, with cell-free channels extending from the surface into the biofilm. This indicated that toluene may penetrate to deeper layers of the biofilm, and consequently P. putida may be actively degrading toluene in all regions of the biofilm. Furthermore, measurements of growth rate-related parameters for P. putida showed reduced rRNA content and cell size (relative to that in a batch culture), indicating that the P. putida population was not degrading toluene at a maximal rate in the biofilm environment. Assuming that the rRNA content reflected the cellular activity, a lower toluene degradation rate for P. putida present in the biofilm could be estimated. This calculation indicated that P. putida was responsible for a significant part (65%) of the toluene degraded by the entire community.

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A lab-scale trickling filter for treatment of toluene-containing waste gas was investigated. The filter performance was investigated for various loads of toluene. Two levels of the gas flow were examined, 322 m d−1 and 707 m d−1. The gas inlet concentrations were varied in the range from 0.6 to 4.0 g m−3. The toluene elimination increased linearly with increasing load, and at maximum load the elimination was 50 g m−3 h−1 (70% purification efficiency). This was in accordance with reported values for toluene removal in trickling filters. The removal was determined by the gas/liquid mass transfer and the biological degradation as well. An analytical model described the toluene removal as a half-order removal by use of two sets of parameters for the gas/liquid mass transfer and the biological degradation due to the two different gas flows. The mass transfer coefficients and the surface removal rates estimated by parameter fitting corresponded to previously observed values. The effect of the gas flow on the mass transfer coefficient and the biological removal rate may be explained by different flow patterns of the gas and the liquid phases. A characterisation of the biofilm showed an almost even biofilm growth over the filter height, which was in accordance with a constant liquid concentration throughout the column.