Tertiary Nitrogen Removal Research Articles

An innovative bilayer solid carbon source for tertiary denitrification: Synthesis, performance, and microbial diversity analysis

The low C/N ratio poses a constraint on the tertiary nitrogen removal process in domestic sewage. Composite solid carbon sources have recently gained attention in recent years for improving nitrogen removal efficacy. However, little research has focused on the lengthy start-up of denitrification systems caused by composite solid carbon sources. In this study, we employed a biodegradable sodium alginate (SA) and corn cob (CC) mixture, implanted a polycaprolactone (PCL) and corn starch (CS) mixture, and created a new composite solid carbon source polycaprolactone-corn starch‑sodium alginate-corn cob microspheres (PCSC). The early rapid carbon release and late stable carbon release properties of PCSC allow it to rapidly enrich denitrifying bacteria and form biofilms on its surface. It only takes 5 days to start denitrification biological filters utilizing PCSC as a carbon source, with an average nitrogen removal rate of 85 %, which is 60 % and 10 % higher than SA-CC and PCL-CS, respectively. The 16S rRNA gene high-throughput sequencing revealed that the primary denitrifying bacteria on the surface of PCSC were Chitinophagaceae, Comamonadaceae, and Blastocatellaceae. Overall, PCSC can make the denitrification system start up fast, steadily, and efficiently, and it has tremendous application prospects in tertiary nitrogen removal of wastewater.

Advanced nitrogen removal in a fixed-bed anaerobic ammonia oxidation reactor following an anoxic/oxic reactor: Nitrogen removal contributions and mechanisms

This study demonstrated the feasibility of anaerobic ammonia oxidation (anammox) served as tertiary nitrogen removal process. An upflow fixed-bed reactor (UFBR) pre-inoculated with anammox bacteria (AnAOB) followed an anoxic/oxic (A/O) reactor treating magnetic-coagulation pretreated municipal wastewater. When bypassing 15% of influent into UFBR, UFBR removed 5.37 mg-TN/L contributing to 23.4% on total TN removal, in which the combination of partial nitritation and partial denitrification with anammox was main nitrogen removal pathway. Relatively low concentrations of NH4+-N and anaerobic environment promoted the growth of ammonia oxidizing archaea (AOA) in the inner-layer of biofilm in UFBR. The cooperation of AOA and ammonia-oxidizing bacteria (AOB) with AnAOB was achieved, with AOA, AOB, and AnAOB abundances of 0.01–0.32%, 0.25–0.44%, and 0.77–2.18% on the biofilm, respectively. Metagenomic analysis found that although AOB was the main NH4+-N oxidizer, archaeal amo gene on biofilm increased threefold during 90 days’ treatment.

Comparison of Polycaprolactone and Poly-3-hydroxybutyrate-co-3-hydroxyvalerate for Nitrogen Removal

Two types of biodegradable polymers, polycaprolactone (PCL) and poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), were used as a denitrification slow-release carbon source and a microbial carrier. By comprehensively comparing their performances in denitrification, carbon release, surface morphology, and material composition as well as their microbial community characteristics, the PHBV was determined as the better performer. It had a shorter denitrification start time, a higher denitrification rate, a lower residual organic matter concentration, and a more stable and sustained denitrification performance than PCL. This is because its surface was rough and contained large amounts of hydrophilic groups such as C-O and C=O, which is easily attached and degraded by microorganisms. As a result, the microorganisms on its surface were diverse. The dominant ones were identified with heterotrophic denitrification potentials, such as Thiothrix, Pseudomonas, Zoogloea, Flavobacterium, and Dechloromonas. Therefore, PHBV is suitable as a carbon source medium for tertiary nitrogen removal.

Methane-based denitrification kinetics and syntrophy in a membrane biofilm reactor at low methane pressure

A methane-based membrane biofilm reactor (MBfR) was assessed for a tertiary nitrogen removal process in domestic wastewater treatment. To mitigate effluent dissolved methane concentrations, the MBfR was operated with a 20% methane mixing ratio and a low pressure of 0.003 atm. The nitrate concentration was reduced from 20 to 4 mg/L with a low methane concentration of 3.3 mg/L in the effluent at 4 h hydraulic retention time (HRT). An in situ dissolved oxygen sensor showed a concentration of 0.045 mg/L in the MBfR, demonstrating methane oxidation under hypoxic conditions. Both 16S rRNA gene sequencing and metagenomic analysis identified bacteria capable of oxidation of methane coupled to denitrification (Methylocystis), whereas other bacteria were implicated in either methane oxidation (Methylococcus) or nitrate reduction (Escherichia). Reduced genetic potential for nitrate reduction to nitrite at a shorter HRT coincided with a decreased efficiency of denitrification, suggesting rate limitation by this initial step of denitrification. Genes encoding nitrite reduction to dinitrogen were at similar relative abundance under both HRT conditions. Our results provide mechanistic evidence for microbial syntrophy between aerobic methanotrophs and denitrifiers in methane-fed MBfRs operated under varying HRTs, with important implications for novel biological nitrogen removal to dilute wastewater.

Effect of temperature on tertiary nitrogen removal from municipal wastewater in a PHBV/PLA-supported denitrification system.

In this study, a poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/poly(lactic acid) (PHBV/PLA)-supported denitrification system was built to remove nitrogen from municipal wastewater treatment plant secondary effluent, and the influence of operating temperature on nitrogen removal was further investigated. Results indicated that a PHBV/PLA-supported denitrification system could effectively fulfill the tertiary nitrogen removal. The nitrogen removal efficiency gradually declined with the operating temperature decreasing, and the denitrification rate at 30°C was 5 times higher than that at 10°C. Meanwhile, it was found that a slight TOC accumulation only occurred at 30°C (with an average of 2.03mg/L) and was avoided at 10~20°C. The reason for effluent TOC variation was further explained through the consumption and generation pathways of TOC in this system. Furthermore, the temperature coefficient was about 0.02919, indicating that the PHBV/PLA-supported denitrification system was a little sensitive to temperature. A better knowledge of the effect of operating temperature will be significant for the practical application of the solid-phase denitrification system.

Novel two stage partial denitrification (PD)-Anammox process for tertiary nitrogen removal from low carbon/nitrogen (C/N) municipal sewage

High nitrate (NO3−-N) concentration in the effluent of wastewater treatment plants (WWTPs), cannot meet the increasing stringent discharge limits. Hence, the tertiary treatment is necessary to lower the total nitrogen concentration. In this study, an innovative partial denitrification (PD)-Anammox process was applied to remove the nitrate nitrogen (20–40 mg N/L) from secondary effluent. The nitrate wastewater were firstly fed to PD sequencing batch reactor (SBR, 10 L) to produce nitrite along with the low carbon/nitrogen ratio (C/N) municipal sewage (NH4+ of 57.8 mg N/L, COD of 175.8 mg/L), then the effluent from SBR was pumped to the anaerobic sludge blanket (UASB, 3.2 L) performing anammox for further nitrogen removal. The integrated process was operated for 224 days with the secondary effluent to municipal sewage volume ratio of 2.9–6. The results suggested that an excellent nitrate removal efficiency of 97.9% was achieved, and the mean removal efficiency of NH4+-N and COD from municipal sewage were 95.2% and 81.6%, respectively, leading to the total nitrogen and COD concentration in the final effluent as low as 4.0 mg N/L and 30.1 mg/L, respectively. Anammox was the main nitrogen removal pathway with a mean proportion of 78.2%, and Candidatus_Brocadia was identified as the dominating genus. Furthermore, it was found that a minor nitrous oxide (N2O) was produced in the integrated process. The PD-Anammox process was verified to be economically and environmentally feasible for retrofitting of existing plants.

Effects of residual organics in municipal wastewater on hydrogenotrophic denitrifying microbial communities

Hydrogenotrophic denitrification is promising for tertiary nitrogen removal from municipal wastewater. To reveal the influence of residual organics in municipal wastewater on hydrogenotrophic denitrifiers, we adopted high-throughput 16S rRNA gene amplicon sequencing to examine microbial communities in hydrogenotrophic denitrification enrichments. Using effluent from a municipal wastewater treatment plant as water source, COD, nitrate and pH were controlled the same except for a gradient of biodegradable carbon (i.e., primary effluent (PE), secondary effluent (SE), or combined primary and secondary effluent (CE)). Inorganic synthetic water (IW) was used as a control. Hydrogenophaga, a major facultative autotroph, accounted for 17.1%, 5.3%, 32.7% and 12.9% of the sequences in PE, CE, SE and IW, respectively, implicating that Hydrogenophaga grew well with or without organics. Thauera, which contains likely obligate autotrophic denitrifiers, appeared to be the most dominant genera (23.6%) in IW and accounted for 2.5%, 4.6% and 8.9% in PE, CE and SE, respectively. Thermomonas, which is related to heterotrophic denitrification, accounted for 4.2% and 7.9% in PE and CE fed with a higher content of labile organics, respectively. In contrast, Thermomonas was not detected in IW and accounted for only 0.6% in SE. Our results suggest that Thermomonas are more competitive than Thauera in hydrogenotrophic denitrification with biodegradable organics. Moreover, facultative autotrophic denitrifiers, Hydrogenophaga, are accommodating to residual organic in effluent wastewater, thus we propose that hydrogenotrophic denitrification is amenable for tertiary nitrogen removal.

Tertiary nitrogen removal using simultaneous partial nitrification, anammox and denitrification (SNAD) process in packed bed reactor

The study aimed to polish wastewater effluent from an activated treatment system for nitrogen removal by a packed bed bioreactor (PBR). The nitrogen removal via simultaneous partial nitrification, anammox and denitrification (SNAD) process was investigated with different hydraulic retention times (HRTs). The results show that both effluent ammonia- and nitrate-nitrogen concentrations approached to 0 mg/L at the HRT as low as 3 h. The total nitrogen removal efficiency decreased from an average of 84% at both 18- and 24-h HRTs to 38% at 3-h HRT. This low nitrogen removal efficiency was due to the presence of incomplete degradation of organic nitrogen at shorter HRTs. The analysis using quantitative real-time polymerase chain reaction resolves that the ratio of ammonia-oxidizing bacteria (AOB) to the domain Eubacteria changed limited in the range between 2.8 and 4.0% during the continuous operation. Meanwhile, the nitrite-oxidizing bacteria (NOB) increased from 0.0 to 2.9% when the HRT was gradually reduced to 3 h. The anammox bacteria accounting for 30.5% of microbial consortia dominated the AOB and the NOB at the 3-h HRT. In sum, inorganic form of nitrogen in a typical wastewater plant effluent could be efficiently polished via SNAD process in the PBR.

Tertiary nitrogen removal for municipal wastewater using a solid-phase denitrifying biofilter with polycaprolactone as the carbon source and filtration medium

Tertiary nitrogen removal technologies are needed to reduce the excess nitrogen that is discharged into sensitive aquatic ecosystems. An integrated solid-phase denitrification biofilter (SDNF) was developed with dual media to remove nitrate and suspended solids (SS) from the secondary effluent of municipal wastewater treatment plants. Biodegradable polymer pellets of polycaprolactone (PCL) served as the biofiltration medium and carbon source for denitrification. Long-term continuous operation of the SDNF was conducted with real secondary effluent to evaluate the denitrification performance and effects of influent nitrate loading rates (NLR) and operating temperatures. The results indicated that both nitrate and SS were effectively removed. The SDNF had a strong tolerance for fluctuations in influent NLR, and a maximum denitrification rate of 3.80 g N/(L·d) was achieved. The low temperature had a significant impact on nitrogen removal, yet the denitrification rate was still maintained at a relative high level to as much as 1.23 g N/(L·d) even at approximately 8.0 °C in winter. Nitrite accumulation and excessive organics residue in the effluent were avoided throughout the whole experiment, except on occasional days in the lag phase. The observed biomass yield was calculated to be 0.44 kgVSS/kgPCL. The microbial diversity and community structure of the biofilm in the SDNF were revealed by Illumina high-throughput sequencing. The special carbon source led to an obvious succession of microbial community from the initial inoculum (activated sludge from aerobic tanks), and included a decrease in microbial diversity and a shift in the dominant groups, which were identified to be members of the family Comamonadaceae in the SDNF. The SDNF developed in this study was verified to be an efficient technology for tertiary nitrogen removal from secondary effluent.

Starch/polyvinyl alcohol blended materials used as solid carbon source for tertiary denitrification of secondary effluent

Starch/polyvinyl alcohol (PVA) blended materials for using as a solid carbon source (SCS) were prepared by blending PVA and gelatinized starch in an aqueous solution system, in which PVA served as framework material and starch as carbon source. The optimization of starch content and temperature effects were investigated. It was indicated that higher denitrification efficiency could be achieved with more starch in the materials. The average specific denitrification rates were 0.93, 0.66, 0.37 and 0.36 mg/(g·day) corresponding to starch content of 70%, 60%, 40% and 30% respectively at 37°C. The denitrification rates increased when operating temperature was raised from 23°C to 30°C and then 37°C. The mechanism of carbon release was analyzed incorporating the experimental results of abiotic release in deionized water. The organic carbon was mainly hydrolyzed by microbes, and the biological release efficiencies were at the range of 89.2% to 96.0%. A long-term experiment with a continuous flow reactor with SCS material containing 70% starch was conducted to gain some experience for practical application. When the influent nitrate concentration was in the range of 35.2 to 39.1 mg/L, hydraulic retention time of 4 hr, and operating temperature of 30°C, a nitrogen removal efficiency up to 94.6% and denitrification rate of 0.217 kg/(m3·day) was achieved. The starch-based materials developed in this study can be used as a solid carbon source for tertiary nitrogen removal from secondary effluent.

Hydrogenotrophic denitrification for tertiary nitrogen removal from municipal wastewater using membrane diffusion packed-bed bioreactor

A lab-scale membrane diffusion packed-bed bioreactor was used to investigate hydrogenotrophic denitrification for tertiary nitrogen removal from municipal wastewater. After start-up, the bioreactor had been operated for 165 days by stepwise increasing influent loading rates at 30 and 15°C. The results indicated that this bioreactor could achieve relatively high nitrogen removal efficiencies. The denitrification rates reached 0.250 and 0.230 kg N/(m(3)d) at 30 and 15°C respectively. The total nitrogen concentration in effluent was entirely below 2.0 mg/L at the steady operation state. The average increase of total organic carbon in effluent was approximately 0.41 mg/L, suggesting the risk of organic residue can be completely controlled. Dissolved oxygen (DO) did not show obviously negative effects on hydrogenotrophic denitrification. There was only slight decrease of DO concentration in effluent, which demonstrated almost all of the hydrogen was used for nitrate reduction.

Anaerobes, aerobes and phototrophs (A winning team for wastewater management)

Current mainstream technologies for wastewater treatment, such as activated sludge and tertiary nutrient removal, are too costly to provide a satisfactory solution for the increasing wastewater problems in developing regions. Besides, these technologies do not allow for re-use of valuable energy and nutrients contained in the wastewater. In light of these limitations, it is important to reconsider the technology and strategic approaches in wastewater management. This paper introduces the "Cleaner Production" concept to sewage management, which combines two approaches: pollution prevention and re-use. Pollution prevention includes a shift towards low water use sanitation technology. The more concentrated wastewater, becomes more attractive for re-use oriented treatment schemes. A combination of anaerobic pre-treatment followed by macrophyte-covered stabilisation ponds is proposed for the effective recovery of energy and nutrients from sewage. By selecting optimal applications of the plant biomass and pond effluent, nutrients will end up as fish and crop protein. This contrasts favourably to tertiary nitrogen removal in activated sludge systems, which recycles ammonia through molecular nitrogen at the expense of energy and high costs. Macrophyte ponds are proposed as a key step in waste recycling, because these form the central unit of a recycling engine, driven by photosynthesis. The process is energy efficient, cost effective and applicable under a wide variety of rural and urban conditions.