Novel anammox granules formation from conventional activated sludge for municipal wastewater treatment through flocs management.

The direct integration of anammox process into municipal wastewater treatment has caused widespread concern, but the lack of anammox seeds limited its real application. This study successfully cultivated anammox granules (322.0 µm) from conventional activated sludge treating municipal wastewater. Through ultra-low floc sludge retention times of 8d, nitrifiers on flocs were eliminated and partial nitrification was realized. Furthermore, highly bacteria-enriched granules were initially formed, with Nitrosomonas and Ca. Competibacter 4-fold higher than that of flocs. Specific staining results revealed the microbial interaction with Ca. Brocadia, considering that Ca. Competibacter and Nitrosomonas correspondingly identified in the inner and outer layers of granules. The percentage of Ca. Brocadia present on the granules increased substantially from 0.0 % to 3.0 %, accompanied by a nitrogen removal rate of 0.3 kg·m-3·d-1. Our findings revealed a valuable reference for the anammox bacteria in-situ enrichment under mainstream conditions, which provides theoretical guidance for anammox-based processes practical application.

Anammox granule destruction and reconstruction in a partial nitrification/anammox system under hydroxylamine stress.

Nitrite oxidizing bacteria (NOB) outcompeting anammox bacteria (AnAOB) poses a challenge to the practical implementation of the partial nitrification/anammox (PN/A) process for municipal wastewater. A granules-based PN/A bioreactor was operated for 260 d with hydroxylamine (NH2OH) added halfway through. qPCR results detected the different amounts of NOB among granules and flocs and the dynamic succession during operation. CLSM images revealed a unique layered structure of granules that NOB located inside led to the inhibition effect of NH2OH delayed. Besides, the physical and morphological characteristics revealed that anammox granules experienced destruction. AnAOB took the broken granules as an initial biofilm aggregate to reconstruct new granules. RT-qPCR and high throughput sequencing results suggested that functional gene expression and community structure were regulated for the AnAOB metabolism process. Correspondingly, the rapid proliferation (0.52 â†’ 1.99%) of AnAOB was realized, and the nitrogen removal rate achieved a nearly quadruple improvement (0.21 â†’ 0.83 kg-N/m3·d). This study revealed that anammox granules can self-reconstruct in the PN/A system when granules are disintegrated under NH2OH stress, broadening the feasibility of applying PN/A process.

Successful Application of Anammox Using the Hybrid Autotrophic-Heterotrophic Denitrification Process for Low-Strength Wastewater Treatment.

Directly integrating anammox into sewage treatment is attractive, but anammox bacteria (AnAOB) enrichment is complex due to vicious competition from heterotrophic bacteria (HB). A novel strategy of optimal organics management using a preanaerobic stage and subsequent limited-oxygen conditions (0.32 ± 0.15 mg-O2/L) is applied, and a hybrid autotrophic-heterotrophic denitrification process is developed to treat sewage-like wastewater with a COD/N ratio of 3.1 for 420 days. The stable process was achieved, and a high total nitrogen removal rate of 0.53 kg-N/(m3·d) was obtained compared to conventional nitrification/denitrification. The 16S rRNA high-throughput sequencing analysis suggested that the relative abundance of the nonendogenous HB (Denitratisoma and Thauera) was drastically reduced (P ≤ 0.001), whereas the endogenous denitrifying HB (Candidatus (Ca.) Competibacter) was significantly enriched in the anammox granules (9.98%, P ≤ 0.001). Moreover, Ca. Competibacter as an inner core and Nitrospira and Ca. Brocadia as an outside coating of the anammox granules indicated the cooperation of AnAOB with HB as revealed by laser-scanning confocal microscopy and qPCR. In situ tests further confirmed nitrite from two pathways (partial nitritation and endogenous partial denitritation) that favored AnAOB enrichment. Optimal organics management can mitigate the competition of HB with AnAOB by redirecting the metabolic pathways and microbial community, which is critical to directly integrating anammox into sewage treatment.