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Nitrification is a key process in nitrogen cycling. Several nitrification rate measure- ment techniques rely on the use of chemical inhibitors that effectively and specifically inhibit nitri- fying bacteria. The discovery of ammonia-oxidizing archaea has raised the question of whether these inhibitors are also effective for archaea. The efficiency of 2 commonly used chemicals — allylthiourea (ATU) and nitrapyrin (commercial name N-serve) — in inhibiting nitrification was tested in natural seawater samples containing archaea. In addition, the effect of these inhibitors was tested in a pure culture of Pseudomonas chlororaphis, a bacterium that is used in the denitri- fier method to reduce nitrate to nitrous oxide, whose isotopic composition can be analyzed with isotopic ratio mass spectrometers. This method is commonly used in stable isotope based nitrifica- tion rate measurements. ATU inhibited nitrification by 80% at a 100 μM concentration, whereas N-serve inhibited nitrification completely at a 150 μM concentration. Although the inhibition of nitrification by ATU was not complete for the natural seawater ammonia-oxidizing community, <100 μM concentration has been shown to be efficient. For N-serve, the previously found effective concentration (150 μM) for the natural ammonia-oxidizing community inhibited nitrification com- pletely. Both ATU and N-serve reduced nitrous oxide production in the denitrifier method, but the 15 N atom% values were not affected. The nitrification rates measured with inhibitors should be interpreted with caution because inhibitors can alter the functioning of bacterial methods used to analyze the nitrification rates.

Effect of nitrification inhibitors on the Baltic Sea ammonia-oxidizing community and precision of the denitrifier method