Interactions between soil, toxicant, and a lux-marked bacterium during solid phase–contact toxicity testing

Abstract

Bioluminescence‐based, solid‐contact toxicityassaysallow test bacterium and toxicant tointeract atthesolid‐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 μg ml−1). 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 > 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.