ANAEROBIC OXIDATION OF AMMONIUM IS A BIOLOGICALLY MEDIATED PROCESS

A newly discovered process by which ammonium is converted to dinitrogen gas under anaerobic conditions (the Anammox process) has now been examined in detail. In order to confirm the biological nature of this process, anaerobic batch culture experiments were used. All of ammonium provided in the medium was oxidized within 9 days. In control experiments with autoclaved or raw wastewater, without added sludge or with added sterilized (either autoclaved or gamma irradiated) sludge, no changes in the ammonium and nitrate concentrations were observed. Chemical reactions could therefore not be responsible for the ammonium conversion. The addition of chloramphenicol, ampicillin, 2,4-dinitrophenol, carbonyl cyanide m-chlorophenyl-hydrazone (CCCP), and mercuric chloride ((HgCl2)-Cl-II) completely inhibited the activity of the ammonium-oxidizing sludge. Furthermore, the rate of ammonium oxidation was proportional to the initial amount of sludge used. It was therefore concluded that anaerobic ammonium oxidation was a microbiological process. As the experiments were carried out in an oxygen-free atmosphere, the conversion of ammonium to dinitrogen gas did not even require a trace of O-2. That the end product of the reaction was nitrogen gas has been confirmed by using (NH4+)-N-15 and (NO3-)-N-14. The dominant product was N-14-15(2). Only 1.7% of the total labelled nitrogen gas produced was N-15-15(2). It is therefore proposed that the N-2 produced by the Anammox process is formed from equimolar amounts of NH4+ and NO3-.