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The temperature dependence of denitrification was investigated for high nitrate nitrogen denitrification in an expanded granular sludge bed (EGSB) reactor. The optimal reaction temperatures were 15–35 °C in which nearly complete denitrification was achieved with the removal of COD maintained over 80%. Nitrite accumulation was observed at 10 °C indicating the incomplete denitrification at low temperature. However, almost complete denitrification was even accomplished as high as 52 °C. High-throughput sequencing detected a total of 84 bacterial genera and 7 phyla, and temperature variation resulted in the shift of microbial community structure and diversity. Proteobacteria thrived while Firmicutes and Bacteroidetes were inhibited by temperature stress. The predominance of Halomonas and the significant decrease of Azoarcus at low temperature indicated a more important role of these two genera in denitrification in an EGSB reactor. The results of qPCR indicated that temperature exerted effects on the abundance of denitrification function genes, nirK, nirS, narG, and nosZ, due to the shift of the bacterial community. This study provided a comprehensive understanding of temperature effects on the denitrification process in an EGSB reactor treating high concentration nitrate wastewater.

rsc advances

Temperature dependence of denitrification microbial communities and functional genes in an expanded granular sludge bed reactor treating nitrate-rich wastewater