With increasing concerns about industrial gas contaminants and the growing demand for durable and sustainable technologies, attentions have been gradually shifted to biological air pollution controls. The ability of Pseudomonas putida PTCC 1694 (bacteria) and Pleurotus ostreatus IRAN 1781C (fungus) to treat contaminated gas stream with toluene and its biological degradation was compared under similar operating conditions. For this purpose, a biofilter on the laboratory scale was designed and constructed and the tests were carried out in two stages. The first stage, bacterial testing, lasted 20 days and the second stage, fungal testing, lasted 16 days. Inlet loading rates (IL) for bacterial and fungal biofilters were 21.62 ± 6.04 and 26.24 ± 7.35 g/m 3 h respectively. In general, fungal biofilter showed a higher elimination capacity (EC) than bacterial biofilter (18.1 ± 6.98 vs 13.7 ± 4.7 g/m 3 h). However, the pressure drop in the fungal biofilter was higher than the bacterial biofilter (1.26 ± 0.3 vs 1 ± 0.3 mm water), which was probably due to the growth of the mycelium. Fungal biofiltration showed a better performance in the removal of toluene from the air stream.