Differences in the Effects of Calcium and Magnesium Ions on the Anammox Granular Properties to Alleviate Salinity Stress

Divalent cations were known to alleviate salinity stress on anammox bacteria. Understanding the mechanism of reducing the salinity stress on anammox granules is essential for the application of the anammox process for saline wastewater treatment. In this study, the effect of Ca2+ and Mg2+ augmentation on the recovery of the activity of freshwater anammox granules affected by salinity stress was evaluated. At the condition of a salinity stress of 5 g NaCl/L, the specific anammox activity (SAA) of the granule decreased to 50% of that of the SAA without NaCl treatment. Augmentation of Ca2+ at the optimum concentration of 200 mg/L increased the SAA up to 78% of the original activity, while the augmentation of Mg2+ at the optimum concentration of 70 mg/L increased the SAA up to 71%. EPS production in the granules was increased by the augmentation of divalent cations compared with the granules affected by salinity stress. In the soluble EPS, the ratio of protein to polysaccharides was higher in the granules augmented by Ca2+ than with Mg2+, and the functional groups of the EPS differed from each other. The amount of Na+ sequestered in the soluble EPS was increased by the augmentation of divalent cations, which seems to contribute to the alleviation of salinity stress. Ca. Kuenenia-like anammox bacteria, which were known to be salinity stress-tolerant, were predominant in the granules and there was no significant difference in the microbial community of the granules by the salinity stress treatment. Our results suggest that the alleviation effect of the divalent cations on the salinity stress on the anammox granules might be associated with the increased production of different EPS rather than in changes to the anammox bacteria.

Effects of Recirculating Aquaculture System Wastewater on Anammox Performance and Community Structure

Recirculating aquaculture systems (RAS) are good candidates for the sustainable development of the aquaculture sector. A current limitation of RAS is the production and accumulation of nitrogenous waste, which could affect fish health. We investigated the potential of the anaerobic ammonia oxidation (anammox) process to treat marine wastewater from a cold-water RAS. We show that the marine anammox bacteria Candidatus Scalindua is a promising candidate. However, its activity was affected by unknown compounds in the RAS wastewater and/or the sub-optimum of essential trace elements (TEs). Anammox activity dropped to 2% and 13% in NH4+ and NO2- removal, respectively, when nitrate-rich RAS wastewater was used as a medium in the absence of TE supplementation. A TE supplementation was added to the RAS wastewater in a subsequent phase, and a recovery in anammox activity was shown (25% and 24% in NH4+ and NO2- removal, respectively). Future studies need to identify the unknown factor and determine the specific needs regarding TE for optimal RAS wastewater treatment by Candidatus Scalindua.

An Analysis of Operation Conditions and Microbial Characteristics in Swine Wastewater Treatment Plants with Spontaneously Enriched Anammox Bacteria

The spontaneous enrichment of anammox bacteria has been reported in swine wastewater treatment facilities. However, their causative conditions and microbial characteristics, which this study aims to explain, are poorly understood. We discovered eight treatment facilities where the collected red biofilms exhibited high anammox activity levels at 57–843 µmol-N2/g-ignition loss (IL)/h and anammox DNA concentrations of 4.3 × 108–1.6 × 1012 copies/g-IL. The facilities used various wastewater treatment methods—six of them employed a multi-stage continuous reactor, whereas aeration tanks were continuously aerated at another combination of six facilities. Levels of dissolved oxygen (DO) in these tanks were fairly low at ≤1 mg/L. Pyrosequencing of the biofilms indicated the presence of 3–62.5% Planctomycetes, and the dominant anammox in each biofilm comprised three operational taxonomic units (OTUs) similar to Candidatus Jettenia asiatica, Ca. Brocadia fulgida, and Ca. B. caroliniensis. This suggested that some particular species of anammox bacteria naturally thrive when operating a swine wastewater treatment facility at low DO levels. The frequent enrichment of anammox biofilms at the sampled sites indicated that these treatment facilities were good seed sources of anammox; therefore, anammox treatment would be a viable method for the removal of nitrogen from swine wastewater.

Simultaneous anaerobic and aerobic ammonium and methane oxidation under oxygen limitation

Methane and ammonia have to be removed from wastewater treatment effluent in order to discharge it to receiving water bodies. A potential solution for this is a combination of simultaneous ammonia and methane oxidation by anaerobic ammonia-oxidizing (anammox) bacteria and nitrite/nitrate-dependent anaerobic methane oxidizing (N-damo) microorganisms. When applied, these microorganisms will be exposed to oxygen but little is known about the effect of a low concentration of oxygen on a culture containing these microorganisms. In this study, a stable co-culture containing anammox and N-damo microorganisms in a laboratory scale bioreactor was established under oxygen limitation. Membrane inlet mass spectrometry (MIMS) was used to directly measure the in situ simultaneous activity of N-damo, anammox and aerobic ammonia oxidizing microorganisms. In addition, batch tests revealed that the bioreactor also harbored aerobic methanotrophs and anaerobic methanogens. Together with FISH analysis and metagenomics, these results indicate that the combination of N-damo and anammox activity under the continuous supply of limiting oxygen concentrations is feasible and can be implemented for the removal of methane and ammonia from anaerobic digester effluents. Importance Nitrogen in wastewater leads to eutrophication of the receiving water bodies and methane is a potent greenhouse gas; it is therefore important that these are removed from wastewater. A potential solution for the simultaneous removal of nitrogenous compounds and methane is the application of a combination of nitrite/nitrate depended methane oxidation (N-damo) and anammox. In order to do so, it is important to investigate the effect of oxygen on these two anaerobic processes. In this study, we investigate the effect of a continuous oxygen supply on the activity of an anaerobic methane and ammonia oxidizing coculture. The findings presented in this study are important for the potential application of these two microbial processes in wastewater treatment.

Start-up Strategies for Anaerobic Ammonia Oxidation (Anammox) in In-Situ Nitrogen Removal from Polluted Groundwater in Rare Earth Mining Areas

The tremendous input of ammonium and rare earth element (REE) ions released by the enormous consumption of (NH4)2SO4 in in situ leaching for ion-adsorption RE mining caused serious ground and surface water contamination. Anaerobic ammonium oxidation (anammox) was a sustainable in situ technology that can reduce this nitrogen pollution. In this research, in situ, semi in situ, and ex situ method of inoculation that included low-concentration (0.02 mg·L−1) and high-concentration (0.10 mg·L−1) lanthanum (La)(III) were adopted to explore effective start-up strategies for starting up anammox reactors seeded with activated sludge and anammox sludge. The reactors were refrigerated for 30 days at 4 °C to investigate the effects of La(III) during a period of low-temperature. The results showed that the in situ and semi in situ enrichment strategies with the addition of La(III) at a low-concentration La(III) addition (0.02 mg·L−1) reduced the length of time required to reactivate the sludge until it reached a state of stable anammox activity and high nitrogen removal efficiency by 60–71 days. The addition of La(III) promoted the formation of sludge floc with a compact structure that enabled it to resist the adverse effects of low temperature and so to maintain a high abundance of AnAOB and microbacterial community diversity of sludge during refrigeration period. The addition of La(III) at a high concentration caused the cellular percentage of AnAOB to decrease from 54.60 ± 6.19% to 17.35 ± 6.69% during the enrichment and reduced nitrogen removal efficiency to an unrecoverable level to post-refrigeration.