Startup, performance, and microbial communities of an anammox reactor inoculated with indigenous sludge for the treatment of high-salinity and mesophilic underground brine
Anaerobic ammonium oxidation (anammox) has been implemented as a cost-effective nitrogen removal process in wastewater treatment. To apply the process to saline wastewater treatment at temperatures below 30 °C, the effectiveness of marine anammox bacteria has been demonstrated in a pilot-scale reactor. Nevertheless, mesophilic conditions, often found in underground brine containing high NH4+ concentrations, have yet to employ an anammox process. The objective of this study was to enrich anammox bacteria capable of removing nitrogen from underground brine possessing a salinity of 3% and at a temperature over 30 °C. To select a promising inoculum, biomass from a brine settling tank in a natural gas plant was subjected to quantifying transcripts of anammox 16S rRNA and hydrazine oxidoreductase (hzo) genes by quantitative reverse transcription polymerase chain reaction. A fixed-bed column anammox reactor was fed with the selected inoculum and operated feeding underground brine mixed with NaNO2 solution at a temperature of 38 °C. As a result, a nitrogen removal rate (NRR) of 1.42 kg-N/m3/day was obtained on day 167. An average NRR of 1.21 kg-N/m3/day and nitrogen removal efficiency of 88% were maintained for 50 days. Amplicon sequencing based on the 16S rRNA revealed that anammox bacteria which are phylogenetically close to Candidatus Kuenenia were successfully enriched in the reactor. These results indicate that nonmarine anammox bacteria can be active and predominant under both high-salinity and mesophilic conditions, making it a likely candidate for effective nitrogen removal from underground and waste brine.
KeywordsAnammox Brine Candidatus Kuenenia Cultivation Temperature
This study was partially funded by the Japanese Society for the Promotion of Science (Grant 17H01893). The authors also thank Professor Barth F. Smets (Technical University of Denmark) and Emeritus Professor Kenji Furukawa (Kumamoto University) for valuable comments.
- Byrne N, Strous M, Crépeau V, Kartal B, Birrien JL, Schmid M, Lesongeur F, Schouten S, Jaeschke A, Jetten M, Prieur D, Godfroy A (2009) Presence and activity of anaerobic ammonium-oxidizing bacteria at deep-sea hydrothermal vents. ISME J 3:117–123. https://doi.org/10.1038/ismej.2008.72 CrossRefGoogle Scholar
- Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Huntley J, Fierer N, Owens SM, Betley J, Fraser L, Bauer M, Gormley N, Gilbert JA, Smith G, Knight R (2012) Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J 6:1621–1624. https://doi.org/10.1038/ismej.2012.8 CrossRefGoogle Scholar
- Kawagoshi Y, Fujisaki K, Tomoshige Y, Yamashiro K, Wei Q (2012) Temperature effect on nitrogen removal performance and bacterial community in culture of marine anammox bacteria derived from sea-based waste disposal site. J Biosci Bioeng 113:515–520. https://doi.org/10.1016/j.jbiosc.2011.11.024 CrossRefGoogle Scholar
- Khramenkov SV, Kozlov MN, Kevbrina MV, Dorofeev AG, Kazakova EA, Grachev VA, Kuznetsov BB, Polyakov DY, Nikolaev YA (2013) A novel bacterium carrying out anaerobic ammonium oxidation in a reactor for biological treatment of the filtrate of wastewater fermented sludge. Microbiology 82:628–636. https://doi.org/10.1134/S002626171305007x CrossRefGoogle Scholar
- Schmid MC, Hooper AB, Klotz MG, Woebken D, Lam P, Kuypers MMM (2008) Environmental detection of octahaem cytochrome c hydroxylamine/hydrazine oxidoreductase genes of aerobic and anaerobic ammonium-oxidizing bacteria. Environ Microbiol 10:3140–3149. https://doi.org/10.1111/j.1462-2920.2008.01732.x CrossRefGoogle Scholar
- Terada A, Zhou S, Hosomi M (2011) Presence and detection of anaerobic ammonium-oxidizing (anammox) bacteria and appraisal of anammox process for high-strength nitrogenous wastewater treatment: a review. Clean Technnol Environ Policy 13:759–781. https://doi.org/10.1007/s10098-011-0355-3 CrossRefGoogle Scholar
- van de Vossenberg J, Rattray JE, Geerts W, Kartal B, van Niftrik L, van Donselaar EG, Damste JSS, Strous M, Jetten MSM (2008) Enrichment and characterization of marine anammox bacteria associated with global nitrogen gas production. Environ Microbiol 10:3120–3129. https://doi.org/10.1111/j.1462-2920.2008.01643.x CrossRefGoogle Scholar
- Yokota N, Watanabe Y, Tokutomi T, Kiyokawa T, Hori T, Ikeda D, Song K, Hosomi M, Terada A (2018) High-rate nitrogen removal from waste brine by marine anammox bacteria in a pilot-scale UASB reactor. Appl Microbiol Biotechnol 102:1501–1512. https://doi.org/10.1007/s00253-017-8663-0 CrossRefGoogle Scholar