Microbial community structure analyses and cultivable denitrifier isolation of Myriophyllum aquaticum constructed wetland under low C/N ratio

With the rapid expansion of livestock production, the amount of livestock wastewater accumulated rapidly. Lack of biodegradable organic matter makes denitrification of livestock wastewater after anaerobic digestion more difficult. In this study, Myriophyllum aquaticum constructed wetlands (CWs) with efficient nitrogen removal performance were established under different carbon/nitrogen (C/N) ratios. Analysis of community composition reveals the change of M. aquaticum CWs in microbial community structure with C/N ratios. The proportion of Proteobacteria which is one of the dominant phyla among denitrifier communities increased significantly under low C/N ratio conditions. Besides, to obtain cultivable denitrifier that could be added into CWs in situ, 33 strains belonging to phylum Proteobacteria were isolated from efficient M. aquaticum CWs, while the best-performing denitrification strain M3-1 was identified as Bacillus velezensis JT3-1 (GenBank No. CP032506.1). Redundancy analysis and quadratic models showed that C/N ratio had significant effects on disposal of nitrate (NO3−-N) and the strains isolated could perform well in denitrification when C/N ratio is relatively low. In addition, they have relatively wide ranges of carbon sources, temperature and a high NO3− removal rate of 9.12 mg/(L·hr) at elevated concentrations of 800 mg/L nitrate. Thus, strains isolated from M. aquaticum CWs with low C/N ratio have a practical application value in the treatment of nitrate-containing wastewater. These denitrifying bacteria could be added to CWs to enhance nitrogen removal efficiency of CWs for livestock wastewater with low C/N ratio in the future.