The enrichment characterisation of Nitrospira under high DO conditions.

Nitrite-oxidizing bacteria (NOB) are crucial to nitrification and nitrogen elimination in wastewater treatment. Mass reports exist on the links between NOB and other microorganisms, for instance, ammonia-oxidizing bacteria (AOB). However, a few studies exist on the enrichment characterisation of NOB under high dissolved oxygen (DO) conditions. In this study, NOB was designed to be enriched individually under high DO conditions in a continuous aeration sequencing batch reactor (SBR), and the kinetic characterisation of NOB was evaluated. The analysis revealed that the average NO2--N removal rate was steady above 98%, with DO and NO2--N being 3-5 mg L-1 and 50-450 mg L-1, respectively. The NO2--N removal efficiency of the system was significantly enhanced and better than in other studies. The high-throughput sequencing suggested that Parcubacteria_ genera_incertae_sedis was the first dominant genus (21.99%), which often appeared in the NOB biological community with Nitrospira. However, the dominant genus NOB was Nitrospira rather than Nitrobacter (8.49%). This result suggested that Nitrospira was capable of higher NO2--N removal. But lower relative abundance indicated that excessive NO2--N had an adverse effect on the enrichment and activity of Nitrospira. In addition, the nitrite half-saturation constant (KNO2) and the oxygen half-saturation constant (KO) were 1.71 ± 0.19 mg L-1 and 0.95 ± 0.10 mg L-1, respectively. These results showed that the enriched Nitrospira bacteria had different characteristics at the strain level, which can be used as a theoretical basis for wastewater treatment plant design and optimisation.

Efficient nitrite accumulation in partial sulfide autotrophic denitrification (PSAD) system: insights of S/N ratio, pH and temperature.

To provide the necessary nitrite for the Anaerobic Ammonium Oxidation (ANAMMOX) process, the effect of nitrite accumulation in the partial sulfide autotrophic denitrification (PSAD) process was investigated using an SBR reactor. The results revealed that the effectiveness of nitrate removal was unsatisfactory when the S/N ratio (mol/mol) fell below 0.6. The optimal conditions for nitrate removal and nitrite accumulation were achieved within the S/N ratio range of 0.7-0.8, resulting in an average Nitrate Removal Efficiency (NRE) of 95.84%±4.89% and a Nitrite Accumulation Rate (NAR) of 75.31%±6.61%, respectively. It was observed that the nitrate reduction rate was three times faster than that of nitrite reduction during a typical cycle test. Furthermore, batch tests were conducted to assess the influence of pH and temperature conditions. In the pH tests, it became evident that the PSAD process performed more effectively in alkaline environment. The highest levels of nitrate removal and nitrite accumulation were achieved at an initial pH of 8.5, resulting in a NRE of 98.30%±1.93% and a NAR of 85.83%±0.47%, respectively. In the temperature tests, the most favourable outcomes for nitrate removal and nitrite accumulation were observed at 22±1 ℃, with a NRE of 100.00% and a NAR of 81.03%±1.64%, respectively. Moreover, a comparative analysis of 16S rRNA sequencing results between the raw sludge and the sulfide-enriched culture sludge sample showed that Proteobacteria (49.51%) remained the dominant phylum, with Thiobacillus (24.72%), Prosthecobacter (2.55%), Brevundimonas (2.31%) and Ignavibacterium (2.04%) emerging as the dominant genera, assuming the good nitrogen performance of the system.

Achieving partial nitrification: A strategy for washing NOB out under high DO condition.

This study investigated the effect of high DO concentrations on PN. The experimental setup involved operating at high DO concentrations (1.5-2.5 mg/L) and environmental temperatures (15-20 °C) over a period of 180 days. Through a sludge enrichment process, the kinetic parameters of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were determined. Surprisingly, contrary to conventional reports, it was observed that NOB exhibited a stronger affinity for DO compared to AOB. As a result, high DO concentrations were necessary to provide favorable conditions for the growth of AOB. In order to establish PN, strategies including intermittent aeration, free ammonia (FA), and controlled sludge retention time (SRT) were employed. The successful PN was achieved with a specific ammonia oxidation rate of 24 mg N/g MLVSS/h and a specific nitrite oxidation rate below 0.10 mg N/g MLVSS/h. The nitrite accumulation rate (NAR) was maintained at 100% during stable operation. The abundance of Nitrosomonas, a typical genus of AOB, was found to be 68.62%, which surpasses previous studies in similar research. A slightly higher DO concentrations may increase energy consumption, but achieve higher efficiency and stability in PN. This study provided new insights into the application of PN in wastewater treatment.

Influencing factors on the activity of an enriched Nitrospira culture with granular morphology.

Nitrospira is a common genus of nitrite-oxidising bacteria (NOB) found in wastewater treatment plants (WWTPs). To identify the key factors influencing the composition of NOB communities, research was conducted using both sequencing batch reactor (SBR) and continuous flow reactor under different conditions. High-throughput 16S rRNA gene sequencing revealed that Nitrospira (18.79% in R1 and 25.77% in R3) was the dominant NOB under low dissolved oxygen (DO) and low nitrite (NO2--N) concentrations, while Nitrobacter (21.26% in R2) was the dominant NOB under high DO and high NO2--N concentrations. Flocculent and granule sludge were cultivated with Nitrospira as the dominant genus. Compared to Nitrospira flocculent sludge, Nitrospira granule sludge had higher inhibition threshold concentrations for free ammonia (FA) and free nitrous acid (FNA). It was more likely to resist adverse environmental disturbances. Furthermore, the effects of environmental factors such as temperature, pH, and DO on the activity of Nitrospira granular sludge were also studied. The results showed that the optimum temperature and pH for Nitrospira granular sludge were 36°C and 7.0, respectively. Additionally, Nitrospira granular sludge showed a higher dissolved oxygen half-saturation constant (Ko) of 3.67 ± 0.71 mg/L due to its morphological characteristics. However, the majority of WWTPs conditions do not meet the conditions for the Nitrospira granular sludge. Thus, it can be speculated that future development of aerobic partial nitrification granular sludge may automatically eliminate the influence of Nitrospira. This study provides a theoretical basis for a deeper understanding of Nitrospira and the development of future water treatment processes.