Metagenomics Response of Anaerobic Ammonium Oxidation (anammox) Bacteria to Bio-Refractory Humic Substances in Wastewater

Anammox-based processes have been widely applied for the treatment of wastewater (e.g., wastewater irrigation systems and constructed wetland) which consists of bio-refractory humic substances. Nonetheless, the impacts of bio-refractory humic substances on anammox consortia are rarely reported. In the present study, three identical lab-scale anammox reactors (i.e., HS0, HS1 and HS10), two of which were dosed with humic substances at 1 and 10 mg·L−1, respectively, were operated for nearly one year. The long-term operation of the reactors showed that the presence of humic substances in influent had no significant influence on nitrogen removal rates. Despite this, comparative metagenomics showed changes in anammox microbiota structure during the exposure to humic substance; e.g., the relative abundance of Candidatus Kuenenia was lower in HS10 (18.5%) than that in HS0 (22.8%) and HS1 (21.7%). More specifically, a lower level of humic substances (1 mg·L−1) in influent led to an increase of genes responsible for signal transduction, likely due to the role of humic substances as electron shuttles. In contrast, a high level of humic substances (10 mg·L−1) resulted in a slight decrease of functional genes associated with anammox metabolism. This may partially be due to the biodegradation of the humic substances. In addition, the lower dosage of humic substances (1 mg·L−1) also stimulated the abundance of hzs and hdh, which encode two important enzymes in anammox reaction. Overall, this study indicated that the anammox system could work stably over a long period under humic substances, and that the process was feasible for leachate treatment.