Microbial diversity reveals the partial denitrification-anammox process serves as a new pathway in the first mainstream anammox plant.

A full-scale sewage treatment plant in Xi'an city is discovered as the first mainstream anaerobic ammonia oxidation (anammox) treatment process in China. Whether its biological mechanism is the nitritation-anammox or partial denitrification (PD)-anammox brought violent controversy between two groups. As a third party, here we uncovered the mystery of the moving-bed biofilm reactor (MBBR) as a PD-anammox process by analyzing the diversity and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) of microbes in anoxic pond. Anammox bacteria was found in the MBBR anoxic tank, which abundance is 8.9 times of that in the common anaerobic-anoxic-oxic process, confirming the existence of anammox process. The denitrifying bacteria (DNB) content in the anoxic tank is 5.9 times of the content of ammonia oxidizing bacteria (AOB), thus the DNB-anammox system is proved at the microbial composition level. The PICRUSt analysis found that ammonium nitrogen is mainly derived from the deamination of urea. The functional genes NAR and AMO of DNB and AOB are 910.84 and 5.80 rpms, respectively. The NAR gene content is 157.0 times of the AMO gene content and it is proved at the genetic level that the nitrite in the anoxic pool is mainly derived from denitrification. This study demonstrated the feasibility and advantages of the PD-anammox in the anammox process, which is different from the traditional nitritation-anammox demonstrated in Strass Wastewater Treatment Plant, Austria and Changi Water Reclamation Plant, Singapore and provided an alternative option for the mainstream application of anammox.

Autoclaved sludge as the ideal seed to culture anammox bacteria: Reactor performance and microbial community diversity.

Reducing activity of commensal bacteria in inocula may enhance anammox bacteria proliferation and realization of anammox process. Fast start-up of anammox process in an UASB reactor was successfully achieved by using autoclaved sludge (anaerobic granular sludge pretreated by autoclaving) and 0.3% active anammox sludge as inoculum. Continuous experiments indicated that R2 (autoclaved sludge addition) could shorten the start-up period from 72days to 63days. The first 50days anammox population specific growth rates (µ) of R1 (the control) and R2 were determined to be 0.014d-1 and 0.045d-1 using q-PCR assays. Analysis of coefficient of variations of nitrogen removal performance during days 96-225 indicated that R2 was more stable than R1. The Illumina MiSeq sequencing showed that autoclaving could decrease microbial diversity of sludge and enhance the abundance of anammox bacteria. Furthermore, PICRUSt community functions forecast and c-di-GMP measure illuminated the result of higher stability in R2.