Identification of nitrogen-incorporating bacteria in a sequencing batch reactor: A combining cultivation-dependent and cultivation-independent method

Abstract Nitrogen-incorporating bacteria in activated sludge play important roles in nitrogen removal in sequencing bactch reactor (SBR), but the active microorganisms and their interactions in the complex community are rarely revealed. Herein, a combining cultivation-dependent and cultivation-independent methods associated with DNA-stable-isotope probing (SIP) was applied to determine the microbes responsible for nitrogen-incorporating in SBR. Results revealed that Cytophagaceae and Sphingobacteriales were identified to be involved in nitrification, and Anaerolineae, Plasticicumulans and Elusimicrobia were responsible for denitrification. Cultivable nitrobacter and denitrifiers were isolated from the activated sludge, but they did not participate in the nitrogen-incorporating based on the SIP results. Additionally, the molecular ecological network analysis indicated that the SIP-identified nitrogen-incorporating bacteria exhibited more links with the intra-community, which might explain the failure of isolating these active bacteria. These findings add understanding of the removal of nitrogenous compounds drived by nitrogen-incorporating bacteria in actual wastewater treatment process.