Department of Civil and Environmental Engineering, Graduate School of Engineering, Hiroshima University, 1–4–1 Kagamiyama, Higashihiroshima 739–8527, Japan.
Microbes Environ. 2011;26(1):67-73. doi: 10.1264/jsme2.me10158.
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 NH(4)(+) and NO(2)(-) 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.
我们使用上流式柱反应器建立了一种富培养海洋厌氧氨氧化（anammox）细菌的方法，该反应器以人工海水为基础，补充氮和矿物质，并接种取自广岛湾的沿海表层沉积物。在反应器运行 2 个月后，观察到同时去除 NH(4)(+)和 NO(2)(-)，表明正在进行 anammox 反应。在第 594 天，达到了 2.17 g-N L(-1) day(-1)的总氮去除率，而氮负荷率为 3.33 g-N L(-1) day(-1)。系统发育分析表明，该反应器中至少存在两种优势的“Candidatus Scalindua”物种。此外，还存在许多未培养的细菌和古菌，包括候选分类或氨氧化古菌。荧光原位杂交（FISH）显示，在第 393 天，anammox 细菌占总细菌的 85.5±4.5%。我们还设计了两种针对每种优势“Candidatus Scalindua”物种的特异性寡核苷酸探针。使用这两种探针同时进行 FISH 分析表明，两种不同的“Candidatus Scalindua”物种在反应器运行期间可以清晰识别并共存，尽管它们的丰度存在一些变化。本研究中富集的海洋 anammox 细菌具有在处理含有高浓度铵盐的工业废水中的应用潜力。
