Effects of toluene on microbially-mediated processes involved in the soil nitrogen cycle

The effects of toluene on indigenous microbial populations involved in the soil nitrogen cycle were examined. Ammonia oxidation potential (AOP) and nitrite oxidation potential (NOP) were both reduced after incubation with high toluene concentrations for 45 days, with the former activity showing greater sensitivity. KCl-extractable ammonium (NH4(+)ext) levels increased dramatically in soil exposed to high toluene levels, and arginine ammonification was not significantly affected. Alfalfa-amended soil incubated in the presence of 200 microgram toluene ml(-1) showed progressive accumulation of NH4(+)ext over 37 days, indicating that mineralization of plant-associated nitrogen was not hindered by toluene. AOP in treated soil was much less than in control soil on days 7 and 37, but the MPN of ammonia oxidizers in control and exposed soil were not significantly different. Soil incubated with 100 microgram toluene ml(-1) for 28 days, vented and allowed to incubate for an additional 7 to 30 days, exhibited only slight increases in AOP and NOP, while NH4+ext returned to control levels within a week. Soil exposed to 200 microgram toluene ml(-1) and treated in the same manner showed no increases in either AOP or NOP, and NH4+ext remained elevated for the duration of the experiment, indicating more long-term effects on soil nitrogen cycling had occurred. Ammonia oxidizer levels in control soil and soil incubated with 100 microgram toluene ml(-1) were not appreciably different, whereas levels of ammonia oxidizers were very low in soil exposed to 200 microgram toluene ml(-1) and increased only slightly by 30 days post vent. Experiments to determine how toluene affects the AOP of soil indicated a competitive inhibition mechanism, with an effective concentration causing 50% reduction in activity (EC50) of 11 micromolar toluene, and a competitive inhibition constant (Ki) of 0.1 +/- 0.05 micromolar toluene. These results indicate the potential for toluene to adversely impact nitrogen cycling in the terrestrial ecosystem by affecting indigenous soil nitrifiers, which are sensitive to lower levels of toluene than has been previously reported.