Despite the implication of anaerobic soil communities in important functions related to C and N biogeochemical cycles, their responses to pesticides are rarely assessed. This study focused on the impact of alachlor, a chloroacetanilide herbicide, on two agricultural soils differing in their land use (fallow and corn-cultivated) in order to investigate the potential adaptation of anaerobic or facultative anaerobic soil microorganisms from fields with long history of herbicide use. The experiment was performed by developing slurries in anoxic conditions over 47 days. Changes in the community structure assessed through terminal restriction fragment length polymorphism analysis of 16S rRNA genes clearly showed a shift in the bacterial community of the cultivated soil, whereas the modification of the microbial community of the fallow soil was delayed. In addition, the analysis of alachlor degradation capacities of the two anaerobic communities indicated that 99 % of alachlor was removed in anoxic slurries of cultivated soil. Both these results suggested the preexistence of microorganisms in the cultivated soil able to respond promptly to the pesticide exposure. The composition of the anaerobic active community determined by 16S rRNA transcript analysis was mainly composed of strictly anaerobic Clostridia and the facultative anaerobe Bacilli classes. Some genera, described for their role in herbicide biodegradation were active in alachlor-treated slurries, whereas others were no longer detected. Finally, this study highlights, when triggered, the important diversity of the anaerobic community in soil.