Excess biomass accumulation and activity loss in vapor-phase bioreactors (VPBs) can lead to unreliable long-term operation. In this study, temporal and spatial variations in biomass accumulation, distribution and activity in VPBs treating toluene-contaminated air were monitored over a 96-day period. Two laboratory-scale bioreactors were subjected to a toluene loading rate of 45.8 g/m(3)-h with one VPB operating in a unidirectional (UD) mode and a second identical VPB operating in a directionally switching (DS) mode. In the UD bioreactor, the contaminated air stream was continuously fed to the bottom of the reactor, while, in the DS bioreactor, the direction of the contaminated gas flow was reversed every three days. Overall, the DS system performed better with respect to biomass distribution and microbial activity across the bioreactor, resulting in more stable bioreactor performance. In contrast, most of the biomass accumulation and activity was confined to the front half of the UD bioreactor column which caused high pressure drops, rapid activity loss and eventually toluene breakthrough. A carbon balance reveals that excess biomass accumulated continuously in both bioreactors, and biomass yield coefficients were very similar (0.59 g dry biomass/g toluene for the UD and 0.63 g dry biomass/g toluene for the DS). The viable biomass population remained relatively constant in both bioreactors over the operational period, while the inactive biomass fraction steadily increased over the same time frame. Biodegradation activity determined by the dehydrogenase enzyme activity assay was found to be a function of biomass accumulation and reflected pollutant removal profiles along the columns. In addition, biomass activity correlated well with the toluene-degrading fraction of the total bacterial population. (C) 2000 John Wiley & Sons, Inc.