Abstract
We established an enrichment culture of marine anaerobic ammonium oxidation (anammox) bacteria using an up-flow column reactor fed with artificial sea water supplemented with nitrogen and minerals and inoculated with coastal surface sediment collected from Hiroshima Bay. After 2 months of reactor operation, simultaneous removal of NH4+ and NO2- was observed, suggesting that an anammox reaction was proceeding. A total nitrogen removal rate of 2.17 g-N L-1 day-1 was attained on day 594 while the nitrogen loading rate was 3.33 g-N L-1 day-1. Phylogenetic analysis revealed that at least two dominant “Candidatus Scalindua” species were present in this reactor. Moreover, many uncultured bacteria and archaea, including candidate division or ammonia-oxidizing archaea, were present. Fluorescence in situ hybridization (FISH) revealed that anammox bacteria accounted for 85.5±4.5% of the total bacteria at day 393. We also designed two oligonucleotide probes specific to each dominant “Candidatus Scalindua” species. A simultaneous FISH analysis using both probes showed that two different “Candidatus Scalindua” species were clearly recognizable and coexisted during reactor operation, although there was some variation in their abundance. The marine anammox bacteria enriched in this study have potential applications to the treatment of industrial wastewater containing high levels of ammonium and salt.
