The overall aim was to identify the microorganisms responsible for carbon uptake from ethylbenzene in soil microcosms using culture-independent methods. Ethylbenzene degradation was observed in only 1 (nitrate-amended agricultural soil) of the 31 different experimental setups tested. To determine which microorganisms were responsible for 13C uptake, stable isotope probing was combined with high-throughput sequencing Illumina amplicon sequencing. For this, total genomic DNA and heavy fraction samples from the live control microcosms (amended with 12C6 ethylbenzene) and sample microcosms (amended with 13C6 ethylbenzene) were subjected to Illumina amplicon sequencing. Several phylotypes were relatively more abundant in the heavy fractions from the 13C6-labeled ethylbenzene-amended soil microcosms compared to the heavy fractions from the live controls. This included unclassified phylotypes within the families Xanthomonadaceae and Rhodocyclaceae. In addition, the total microbial community was characterized in six ethylbenzene degrading microcosms and was dominated by the genus Rhodanobacter (relative abundance 11.7–15.6%), although this phylotype did not illustrate 13C uptake. Azoarcus, a known ethylbenzene degrader, was dominant in only two of the six microcosms. The work indicates that microorganisms not previously linked to ethylbenzene degradation have significant roles in the carbon uptake from this contaminant.
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