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Fenitrothion, a toxic organophosphorus pesticide, can build up the concentration of nitrophenolic compound in soils and hence needs to be removed. Burkholderia sp. FDS-1, a fenitrothion-degrading strain, was used in this work to study factors affecting its growth, and then evaluated for its capacity to degrade fenitrothion in soil microcosms. Minimal salt medium containing 1% (w/v) glucose was found to be a suitable carbon source for inoculum preparation. Various factors, including soil pH, temperature, initial fenitrothion concentration, and inoculum size influenced the degradation of fenitrothion. Microcosm studies performed with varying concentrations (1–200 mg kg−1) of fenitrothion-spiked soils showed that strain FDS-1 could effectively degrade fenitrothion in the range of 1–50 mg kg−1 soil. The addition of Burkholderia sp. FDS-1 at 2×106 colony forming units g−1 soil was found to be suitable for fenitrothion degradation over a temperature range of 20–40 °C and at a slight alkaline pH (7.5). The results indicate that strain FDS-1 has potential for use in bioremediation of fenitrothion and its metabolite-contaminated sites. This is a model study that could be used for decontamination of sites contaminated with other compounds.