Contamination of wastewater with organic-limited nitrates has become an urgent problem in wastewater treatment. The cooperating heterotrophic with sulfur autotrophic denitrification is an alternative process and the efficiency has been assessed in many studies treating simulated wastewater under different operating conditions. However, due to the complex and diverse nature of actual wastewater, more studies treating actual wastewater are still needed to evaluate the feasibility of collaborative denitrification. In this study, lab-scale experiments were performed with actual nitrate polluted water of two different concentrations, with glucose and sodium thiosulfate introduced as mixed electron donors in the coupling sulfur-based autotrophic and heterotrophic denitrification. Results showed that the optimum denitrification performance was exhibited when the influent substrate mass ratio of C/N/S was 1.3/1/1.9, with a maximum denitrification rate of 3.52 kg
day) and nitrate removal efficiency of 93% in the coupled systems. Illumina high-throughput sequencing analysis revealed that autotrophic, facultative, and heterotrophic bacteria jointly contributed to high nitrogen removal efficiency. The autotrophic denitrification maintained as the predominant process, while the second most prevalent denitrification process gradually changed from heterotrophic to facultative with the increase of influent concentration at optimum C/N/S ratio conditions. Furthermore, the initiation of dissimilatory nitrate reduction to ammonium (DNRA) was very pivotal in promoting the entire denitrification process. These results suggested that sulfur-based autotrophic coupled with heterotrophic denitrifying process is an alternative and promising method to treat nitrate containing wastewater.
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