Bacterial whole-cell biosensors (BWBs) have unique advantages over conventional environmental monitoring techniques on the detection of toxicity and bioavailability of contaminants in water and soils. BWBs can also be rapid, sensitive, semiquantitative, cost-effective, and easy to use. In this study, a standard method is described for the detection of contaminants and toxicity in real water and soil samples using Acinetobacter baylyi ADP1-based biosensors.