Successful application of nitritation/anammox to wastewater with elevated organic carbon to ammonia ratios

• Nitritation/anammox with high influent COD/N ratio (acetate and glucose addition).
• SRT is the key for successful AMX competition with HET at high influent COD/N ratio.
• SRT decrease due to a change of the floc morphology caused washout of AMX.
• Dominance of “Candidatus Brocadia fulgida” indicated acetate utilisation by AMX.

The nitritation/anammox process has been mainly applied to high-strength nitrogenous wastewaters with very low biodegradable organic carbon content (<0.5 g COD∙g N−1). However, several wastewaters have biodegradable organic carbon to nitrogen (COD/N) ratios between 0.5 and 1.7 g COD∙g N−1 and thus, contain elevated amounts of organic carbon but not enough for heterotrophic denitrification. In this study, the influence of elevated COD/N ratios was studied on a nitritation/anammox process with suspended sludge. In a step-wise manner, the influent COD/N ratio was increased to 1.4 g COD∙g N−1 by supplementing digester supernatant with acetate. The increasing availability of COD led to an increase of the nitrogen removal efficiency from around 85% with pure digester supernatant to >95% with added acetate while the nitrogen elimination rate stayed constant (275 ± 40 mg N∙L−1∙d−1). Anammox activity and abundance of anammox bacteria (AMX) were strongly correlated, and with increasing influent COD/N ratio both decreased steadily. At the same time, heterotrophic denitrification with nitrite and the activity of ammonia oxidising bacteria (AOB) gradually increased. Simultaneously, the sludge retention time (SRT) decreased significantly with increasing COD loading to about 15 d and reached critical values for the slowly growing AMX. When the SRT was increased by reducing biomass loss with the effluent, AMX activity and abundance started to rise again, while the AOB activity remained unaltered. Fluorescent in-situ hybridisation (FISH) showed that the initial AMX community shifted within only 40 d from a mixed AMX community to “Candidatus Brocadia fulgida” as the dominant AMX type with an influent COD/N ratio of 0.8 g COD∙g N−1 and higher. “Ca. Brocadia fulgida” is known to oxidise acetate, and its ability to outcompete other types of AMX indicates that AMX participated in acetate oxidation. In a later phase, glucose was added to the influent instead of acetate. The new substrate composition did not significantly influence the nitrogen removal nor the AMX activity, and “Ca. Brocadia fulgida” remained the dominant type of AMX. Overall, this study showed that AMX can coexist with heterotrophic bacteria at elevated influent COD/N ratios if a sufficiently high SRT is maintained.

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