In the biological pretreatment of landfill leachate in Mechernich (Germany) a loss of inorganic nitrogen of up to 90% was observed in the nitrification step (rotating biological contactor) under low DO conditions. Ammonia was removed but only small amounts of nitrate were produced. Nitrite accumulation did not occur. In aerobic batch tests nitrogen loss was confirmed without any addition of organic substrate, even when homogenizing the biofilm mechanically to destroy possible anoxic microzones. N2 was measured as the gaseous end product of the process. From results presented it can be assumed that a largely autotrophic microorganism population performed this aerobic nitrification/denitrification.
In full scale wastewater treatment plants with at times considerable deficits in the nitrogen balances, it could hitherto not be sufficiently explained which reactions are the cause of the nitrogen losses and which micro-organisms participate in the process. The single stage conversion of ammonium into gaseous end-products--which is henceforth referred to as deammonification--occurs particularly frequently in biofilm systems. In the meantime, one has succeeded to establish the deammonification processes in a continuous flow moving-bed pilot plant. In batch tests with the biofilm covered carriers, it was possible for the first time to examine the nitrogen conversion at the intact biofilm. Depending on the dissolved oxygen (DO) concentration, two autotrophic nitrogen converting reactions in the biofilm could be proven: one nitritation process under aerobic conditions and one anaerobic ammonium oxidation. With the anaerobic ammonium oxidation, ammonium as electron donor was converted with nitrite as electron acceptor. The end-product of this reaction was N2. Ammonium and nitrite did react in a stoichiometrical ratio of 1:1.37, a ratio which has in the very same dimension been described for the ANAMMOX-process (1:1.31 +/- 0.06). Via the oxygen concentration in the surrounding medium, it was possible to control the ratio of nitritation and anaerobic ammonium oxidation in the nitrogen conversion of the biofilm. Both processes were evenly balanced at a DO concentration of 0.7 mg/l, so that it was possible to achieve a direct, almost complete elimination of ammonium without addition of nitrite. One part of the provided ammonium did participate in the nitritation, the other in the anaerobic ammonium oxidation. Through the aerobic ammonium oxidation into nitrite within the outer oxygen supplied layers of the biofilm, the reaction partner was produced for the anaerobic ammonium oxidation within the inner layers of the biofilm.
In a biological contactor that is part of the biological pretreatment of landfill leachate in Mechernich (Germany) nitrogen elimination of 60% or more was observed under low dissolved oxygen (DO) conditions. Ammonia was converted without accumulation of nitrite and with only little nitrate production. Interestingly, due to limited supply with organic substrate in the system, this observation cannot simply be explained by a combination of conventional autotrophic nitrification and heterotrophic denitrification. In situ hybridization with 16S rRNA-targeted probes revealed the presence of large microcolonies of at least three different types of ammonia-oxidizing bacteria in those biofilm regions where extremely high nitrogen losses occurred. These results were confirmed by comparative sequence analysis of biofilm-derived amoA (encoding the active-site polypeptide of ammonia-monooxygenase) clones for molecular fine-scale analysis of the ammonia-oxidizing population. In batch tests inoculated with biofilm material nitrogen loss occurred without dosage of organic substrate at a DO concentration of 1 mg/l. The simultaneous presence of ammonia and nitrite in the reactor induced the process of complete nitrogen elimination. N2 was identified to be the gaseous end product of the reaction. These results indicate that under low DO concentrations autotrophic ammonia-oxidizers might be the causative agents of the observed nitrogen loss by performing aerobic/anoxic denitrification with nitrite as electron acceptor and ammonia (or perhaps hydroxylamin) as electron donor.
Vertical-flow reed beds (VF) with intermittent feeding are extremely reliable regarding aerobic processes. For a save operation with high nitrification rates and without soil clogging it is essential to preserve aerobic conditions in the filter. The challenge is to keep aerobic conditions in the filter without oversizing the system (economical aspects). It is very difficult to determine the current oxygen content in the filters because it ultimately results from complex interactions of a large number of different influencing parameters such as loading rate, degree of clogging, temperature, and hydraulic behaviour of the reed bed. To gain better knowledge of this complex system, different tests and examinations were carried out over several years. Focusing on the questions of identification and the description of conversion and transport processes (water/gas), a full-scale treatment plant under clogged and non-clogged conditions was investigated in detail. Additionally soil column test were carried out. The results make it possible to describe some of the processes and their interactions in the filter body. Recommendations for a safe and controlled operation can be derived.
For the development of alternative concepts for the cost effective treatment of wastewaters with high ammonium content and low C/N-ratio, autotrophic consortia of micro-organisms with the ability to convert ammonium directly into N2 are of particular interest. Several full-scale industrial biofilm plants eliminating nitrogen without carbon source for years in a stable process, are suspected for some time to harbor active anaerobic ammonium oxidizers in deeper, oxygen-limited biofilm layers. In order to identify the processes of the single-stage nitrogen elimination (deammonification) in biofilm systems and to allocate them to the responsible micro-organisms, a deammonifying moving-bed pilot plant was investigated in detail. 15N-labelled tracer compounds were used as well as 16S rDNA libraries and in situ identification of dominant organisms. The usage of rRNA-targeted oligonucleotide probes (FISH) was particularly emphasized on the ammonium oxidizers of the beta-subclass of Proteobacteria and on the members of the order Planctomycetales. The combined application of these methods led to a deeper insight into the population structure and function of a deammonifying biofilm.
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