Monitoring and quantification of organohalide respiring bacteria is essential for optimization of on-site bioremediation of anoxic subsurface sites contaminated with chloroethenes. Molecular monitoring and model simulations were applied to determine degradation performance of an in situ dechlorinating bioreactor and its influence on the contamination plume. Dehalococcoides was the dominant dechlorinating microorganism as revealed by qPCR targeting 16S rRNA- and chloroethene reductive dehalogenase-encoding genes (tceA, vcrA, bvcA). The presence of all three reductive dehalogenases genes indicated coexistence of several distinct organohalide respiring bacterial populations in the bioreactor and groundwater. Mass balancing revealed that main dechlorinating activities were reduction of cis-dichloroethene and vinyl chloride. Analysis of growth kinetics showed that when performance of the bioreactor improved due to especially the addition of molasses, dechlorinating microorganisms were growing close to their maximum growth rate. Once near-complete dehalogenation was achieved, Dehalococcoides only grew slowly and population density did not further increase. The bioreactor influenced dechlorinating populations in the plume with subsequent decrease in chlorinated compound concentrations over time. In the present study, a combination of molecular diagnostics with mass-balancing and kinetic modeling improved insight into organohalide respiring bacteria and metabolite dynamics in an in situ dechlorinating bioreactor and showed its utility in monitoring bioremediation.