Abstract-Sediment and groundwater samples obtained from a benzene, toluene, ethylbenzene, and xylene (BTEX)-contaminated site were screened, using a bioluminescence-based (lux gene-marked) bacterial biosensor, to identify constraints to site remediation. Through a series of sample manipulations and linked biosensor responses, constraints to BTEX remediation such as adverse pH, presence of nonvolatile organic contaminants, and the presence of heavy metals were investigated. Conventional chemical analysis (gas chromatography, inductively coupled plasma mass spectroscopy) was used to confirm the reliable performance of the biosensor and to identify its potential contribution to site management to ensure effective remediation. The toxicity results of the biosensor were expressed in % maximum bioluminescence calculated against a blank of double deionized water with pH adjusted to 5.5. Untreated samples caused reductions in percentage bioluminescence from 10 to 95%. Water sample W1, containing a total BTEX concentration of 30,595 g/L caused the highest decrease in bioluminescence (5.78%). This toxicity was significantly reduced after elimination of volatile organic compounds (VOCs) (42.01%) and bioluminescence was further increased to 87.80% after removal of total organic matter. This suggested that VOCs were not the only toxic organopollutants present. Water samples giving high values of bioluminescence contained BTEX concentrations of 17 to 31 g/L.
Abstract-Bioluminescence-based, solid-contact toxicity assays allow test bacterium and toxicant to interact at the solid-solution interface. A lux-marked bacterium, Burkholderia sp. RASC, and 2,4-dichlorophenol (2,4-DCP) were used to characterize these interactions. In the basic bioassay, cells were added to soil slurries containing 2,4-DCP (0-120 g ml Ϫ1 ). After 15 min, soil was removed by centrifugation, and bioluminescence in the supernatant was determined. Investigation of 2,4-DCP adsorption to soil revealed that sorption was linear and not significantly (p Ͼ 0.1) affected by the presence of Burkholderia cells. The numbers of culturable Burkholderia cells in the assay supernatant were 48.2 to 64.8% of the inoculum and independent of the soil weight. The effect of soil on 2,4-DCP toxicity was investigated by comparing soil aqueous extract and contact assays. The percentage bioluminescence for the contact assay was consistently higher than the extract assay at all test concentrations, and counts of viable Burkholderia cells were enhanced by the presence of 2,4-DCP in the contact assay. Expressing results as specific bioluminescence decreased the variability in response and the discrepancy in results between the two protocols. We suggest that solid-contact assays need improvement to ensure defined contact between cells and solid phase, and that the reporting of specific activity should be emphasized.
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