Dehalococcoides sequences from continously fed TCE columns. groundwater metagenome

Trichloroethylene (TCE) contamination of groundwater is a major concern for human health. The aim of this study was to identify the variations in the Dehalococcoides sp. establish across different stages of TCE dehalogenating systems. Eight large columns (1.8 m) were filled with the sediment material from Hill Air force Base, Utah, and continuously fed with the TCE for about seven years. Columns were biostimulated with the whey, Newman Zone® nonionic surfactant emulsified oil + lactate (EOLN), Newman Zone® standard surfactant emulsified oil + lactate (EOL) and un-amended controls, and were established under different stages of TCE reduction ranging from complete, partial and no TCE dehalogenation, respectively. Most of the TCE reduction activity was noticed in the top layers (0-10 cm). Total microbial community as well as variation in the Dehalococcoides sp. was measured through high throughput sequencing. Dehalococcoides sequencing was performed through nested PCR approach. Dehalococcoides 16S rRNA gene was amplified using Dehalococcoides sp. specific primers (F-Dhc1 & R-Dhc2012). The product of first PCR was cleaned and used in the nested PCR reaction. Second PCR was performed using the universal 16S rRNA gene sequencing primers (F515 and R907).