Homogeneous inoculation vs. microbial hot spots of isolated strain and microbial community: What is the most promising approach in remediating 1,2,4-TCB contaminated soils?

We investigated the effectiveness of different inoculation approaches in enhancing the mineralization of [U-14C] labeled 1,2,4-trichlorobenzene (1,2,4-TCB) in soil. Inoculation was conducted with a soil-borne 1,2,4-TCB mineralizing microbial community (MC) as well as the Bordetella sp. strain F2 originally isolated from this community as the key degrader organism (IS). Both were applied either via liquid medium (LM) or attached on clay particles (CP). Fluorescence in-situ hybridization in combination with 14C-1,2,4-TCB mineralization measurements as well as measurements of 14C-residues in soil were used to investigate the bioaugmentation efficiency of the different approaches. Bordetella sp. cell numbers increased about 2-5 times during the incubation process, indicating that the bacteria could survive and develop in the new soil habitat. While the native soil showed negligible 1,2,4-TCB mineralization rates, soil inoculated with the MC attached on CP showed the highest 1,2,4-TCB mineralization rate per Bordetella cell, whereas the other inoculum approaches showed an increased but lower contaminant mineralization. Additionally, the MC-CP approach showed the highest cumulative 1,2,4-TCB mineralization as well as the highest formation of bound 14C-residues which is most likely equivalent to 14C incorporated into the microbial biomass. Thus, our results allow the conclusion that the application of a specific microbe-clay-particle-complex is the most promising approach for an accelerated in-situ mineralization of chemicals in agricultural soils.