Simultaneous nitrification, denitrification, and phosphorus removal from nutrient-rich industrial wastewater using granular sludge

The biological removal of nitrogen and phosphorus from nutrient-rich abattoir wastewater using granular sludge has been investigated. A lab-scale sequencing batch reactor, seeded with granular sludge developed using synthetic wastewater, was operated for 13 months under alternating anaerobic and aerobic conditions. It is demonstrated that the granules could be sustained and indeed further developed with the use of abattoir wastewater. The organic, nitrogen, and phosphorus loading rates applied were 2.7 gCOD L(-1) day(-1), 0.43 gN L(-1) day(-1), and 0.06 gP L(-1) day(-1), respectively. The removal efficiency of soluble COD, soluble nitrogen and soluble phosphorus were 85%, 93%, and 89%, respectively. However, the high suspended solids in the effluent limited the overall removal efficiency to 68%, 86%, and 74% for total COD, TN, and TP, respectively. This good nutrient removal was achieved through the process known as simultaneous nitrification, denitrification, and phosphorus removal, likely facilitated by the presence of large anoxic zones in the center of the granules. The removal of nitrogen was likely via nitrite optimizing the use of the limited COD available in the wastewater. Accumulibacter spp. were found to be responsible for most of the denitrification, further reducing the COD requirement for nitrogen and phosphorus removal. Mineral precipitation was evaluated and was not found to significantly contribute to the overall nutrient removal. It is also shown that the minimum HRT in a granular sludge system is not governed by the sludge settleability, as is the case with floccular sludge systems, but likely by the limitations associated with the transfer of substrates in granules.     

Volatile fatty acids production from food wastes and its application to biological nutrient removal

Korean food wastes were anaerobically digested to produce volatile fatty acids (VFA) that can be used as a carbon source in biological nutrient removal in a sequential batch reactor (SBR). Acetate, propionate and butyrate were produced at a yield of 379-400 g VFA/kg VS₀ (initial volatile solids). The ratio of SCOD (Soluble Chemical Oxygen Demand) of VFA to ammonia nitrogen (N) was in the range of 36.2-36.5 and the ratio of SCOD to phosphorus was between 151 and 162. The removal rate of nitrogen and phosphorus improved from 44% and 37% to 92% and 73%, respectively when the VFA were added to the influent of the Taejon municipal wastewater plant. The concentration of nitrogen and phosphorus were maintained below 3 mg/l and 1 mg/l, respectively. The N- and P-content of the food waste was low enough not to influence the final N- and P-concentrations of the wastewater.     

Improvement in post-treatment of digested swine wastewater

The performance of sequencing batch reactor (SBR) during post-treatment of digested effluent of swine wastewater was investigated. While operating SBR to treat the digested effluent directly, the performance was very poor with COD removal rate about 10%, and NH4+-N removal rate nearly 50%, with a scarce removal of total phosphorus. The performance apparently improved after adding raw swine wastewater or alkali to digested effluent. Although similar results for NH4+-N removal were achieved adopting both measures, the addition of raw wastewater proved superior in removing total nitrogen and total phosphorus. The addition of raw wastewater obtained effluent COD around 300 mg/L which was lower than that after alkali addition i.e. around 550 mg/L. Judged from the investment, oxygen demand, sludge yield, biogas production and running cost, the traditional combined anaerobic-SBR process is unfeasible to treat swine wastewater, while the combined anaerobic-SBR process with addition of raw swine wastewater can be a suitable biotechnology.     

Application of biochemical oxygen demand (BOD) biosensor for optimization of biological carbon and nitrogen removal from synthetic wastewater in a sequencing batch reactor system

A bench scale reactor using a sequencing batch reactor process was used to evaluate the applicability of biosensors for the process optimization of biological carbon and nitrogen removal. A commercial biochemical oxygen demand (BOD) biosensor with a novel microbial membrane was used to determine the duration of each phase by measuring samples in real time in an SBR cycle with filling/anoxic-anaerobic/aerobic/sludge wasting/settling/withdrawal periods. Possible strategies to increase the efficiency for the biological removal of carbon and nitrogen from synthetic wastewater have been developed. The results show that application of a BOD biosensor enables estimation of organic carbon, in real time, allowing the optimization or reduction the SBR cycle time. Some typical consumption patterns for organic carbon in the non-aeration phase of a typical SBR operation were identified. The rate of decrease of BOD measured using a sensor BOD, was the highest in the initial glucose breakdown period and during denitrification. It then slowed down until a 'quiescent period' was observed, which may be considered as the commencement of the aeration period. Monitoring the BOD curve with a BOD biosensor allowed the reduction of the SBR cycle time, which leads to an increase in the removal efficiency. By reducing the cycle time from 8 to 4 h cycle, the removal efficiencies of nitrate, glucose, and phosphorus in a given time interval, were increased to nearly double, while the removal of nitrogen ammonium was increased by one-third.     

Hydrolyzed molasses as an external carbon source in biological nitrogen removal

Hydrolyzed molasses was evaluated as an alternative carbon source in a biological nitrogen removal process. To increase biodegradability, molasses was acidified before thermohydrolyzation. The denitrification rate was 2.9-3.6 mg N/g VSSh with hydrolyzed molasses, in which the percentage of readily biodegradable substrate was 47.5%. To consider the hydrolysate as a carbon source, a sequencing batch reactor (SBR) was chosen to treat artificial municipal wastewater. During the 14 days (28 cycles) of operation, the SBR using hydrolyzed molasses as a carbon source showed 91.6 +/- 1.6% nitrogen removal, which was higher than that using methanol (85.3 +/- 2.0%). The results show that hydrolyzed molasses can be an economical and effective external carbon source for the nitrogen removal process.     

Evaluation of the feasibility of alcohols serving as external carbon sources for biological phosphorus removal induced by the oxic/extended‐idle regime

Recently, a novel operational regime (i.e., the oxic/extended‐idle [OEI] regime) has been reported to successfully achieve enhanced biological phosphorus removal (EBPR) when employing glucose and volatile fatty acids as the sole substrate. In the OEI regime, polyphosphate accumulating organisms (PAOs) could get a selective advantage over other populations during the extended‐idle period where polyphosphate released but polyhydroxyalkanoates and glycogen transformations were negligible/low, thus energy requirements for maintenance purposes in the period could be covered by polyphosphate release. This study further evaluated the feasibility of alcohols as external carbon sources for EBPR induced by the OEI regime, as the available substrate in the raw wastewater is often deficient. First, phosphorus removal in the OEI process was compared, respectively, with methanol and ethanol as the sole substrate. The results showed that the ethanol‐reactor achieved 90.8 ± 2.3% of phosphorus removal, which was approximate twofold than the methanol‐reactor. Further studies displayed that the cells in the ethanol‐reactor contained more PAOs, and had higher activities of exopolyphosphatase and polyphosphate kinase than those in the methanol‐reactor. Also, the aerobic transformations of polyhydroxyalkanoates and glycogen in the ethanol‐reactor were, respectively, higher and lower than those in the methanol‐reactor, which were consistent with the reactors performances. Then, the feasibility of using ethanol as external substrate to enhance EBPR in the OEI process was confirmed for a municipal wastewater. Finally, EBPR performance and metabolic transformation values between the OEI and the anaerobic/oxic (A/O) regimes with ethanol as the sole substrate were compared. The results showed that EBPR in the ethanol‐OEI reactor was higher than that in the ethanol‐A/O reactor. All the above results proved that ethanol was a favorable external substrate to the OEI regime for EBPR enhancement. Biotechnol. Bioeng. 2013; 110: 827–837. © 2012 Wiley Periodicals, Inc.     

Sequencing batch membrane biofilm reactor for simultaneous nitrogen and phosphorus removal: novel application of membrane-aerated biofilm

A sequencing batch membrane biofilm reactor (SBMBfR) was developed for simultaneous carbon, nitrogen, and phosphorus removal from wastewater. This reactor was composed of two functional parts: (1) a gas-permeable membrane on which a nitrifying biofilm formed and (2) a bulk solution in which bacteria, mainly denitrifying polyphosphate-accumulating organisms (DNPAOs), were suspended. The reactor was operated sequentially under anaerobic condition and then under membrane aeration condition in one cycle. During the anaerobic period, organic carbon was consumed by DNPAOs; this was accompanied by phosphate release. During the subsequent membrane aeration period, nitrifying bacteria utilized oxygen supplied directly to them from the inside of the membrane. Consequently, the nitrite and nitrate products diffused into the bulk solution, where they were used by DNPAOs as electron acceptors for phosphate uptake. In a long-term sequencing batch operation, the mean removal efficiencies of total organic carbon (TOC), total nitrogen (T-N), and total phosphorus (T-P) under steady-state condition were 99%, 96%, and 90%, respectively. In addition, fluorescence in situ hybridization (FISH) clearly demonstrated the difference in bacterial community structure between the membrane biofilm and the suspended sludge: ammonia-oxidizing bacteria belonging to the Nitrosomonas group were dominant in the region adjacent to the membrane throughout the operation, and the occupation ratio of the well-known polyphosphate-accumulating organism (PAO) Candidatus "Accumulibacter phosphates" in the suspended sludge gradually increased to a maximum of 37%.     

The investigation of effect of organic carbon sources addition in anaerobic–aerobic (low dissolved oxygen) sequencing batch reactor for nutrients removal from wastewaters

The effect of addition of organic carbon sources (acetic acid and waste activated sludge alkaline fermentation liquid) on anaerobic–aerobic (low dissolved oxygen, 0.15–0.45 mg/L) biological municipal wastewater treatment was investigated. The results showed that carbon source addition affected not only the transformations of polyhydroxyalkanoates (PHA), glycogen, nitrogen and phosphorus, but the net removal of nitrogen and phosphorus. The removal efficiencies of TN and TP were, respectively, 61% and 61% without organic carbon source addition, 81% and 95% with acetic acid addition, and 83% and 97% with waste activated sludge alkaline fermentation liquid addition. It seems that the alkaline fermentation liquid of waste biosolids generated in biological wastewater treatment plant can be used to replace acetic acid as an additional carbon source to improve the anaerobic–aerobic (low dissolved oxygen) municipal wastewater nutrients removal although its use was observed to cause a slight increase of effluent BOD and COD concentrations.     

Denitrifying phosphorus removal and impact of nitrite accumulation on phosphorus removal in a continuous anaerobic-anoxic-aerobic (A2O) process treating domestic wastewater

A lab-scale anaerobic-anoxic-aerobic (A(2)O) process was operated to investigate denitrifying phosphorus removal and nitritation-denitritation from domestic wastewater, especially regarding the impact of nitrite accumulation caused by nitritation on phosphorus removal. The results showed that mean total nitrogen (TN) removal was only about 47% and phosphorus removal was almost zero without the pre-anoxic zone and additional carbon source. Contrastively, with configuration of pre-anoxic zone, TN and phosphorus removal was increased to 75% and 98%, respectively, as well as denitrifying phosphorus removal of 66-91% occurred in the anoxic zone. Nitritation-denitritation was achieved through a combination of short aerobic actual hydraulic retention time and low dissolved oxygen levels (0.3-0.5 mg/L); however, phosphorus removal deteriorated with increase of nitrite accumulation rates. The free nitrous acid (FNA) concentration of 0.002-0.003 mg HNO(2)-N/L in the aerobic zone inhibited phosphorus uptake, which was major cause of phosphorus removal deterioration. Through supplying the carbon sources to enhance denitrification and anaerobic phosphorus release, nitrite and FNA concentrations in the aerobic zone were reduced, and phosphorus removal was improved. Compared with nitrification-denitrification, nitritation-denitritation reduced the carbon requirement by 30% and performed biological nutrients removal well with mean TN and phosphorus removal of 85% and 96%, respectively.     

Simultaneous removal of phosphorus and nitrogen in a sequencing batch biofilm reactor with transgenic bacteria expressing polyphosphate kinase

To improve phosphorus removal from wastewater, we constructed a high-phosphate-accumulating microorganism, KTPPK, using Pseudomonas putida KT2440 as a host. The expression plasmid was constructed by inserting and expressing polyphosphate kinase gene (ppk) from Microcystis aeruginosa NIES-843 into broad-host-range plasmid, pBBR1MCS-2. KTPPK was then added to a sequencing batch biofilm reactor (SBBFR) using lava as a biological carrier. The results showed that SBBFR with KTPPK not only efficiently removed COD, NH(3)-N, and NO(3)(-)-N but also had a high removal capacity for PO(4)(3-)-P, resulting in a low phosphorus concentration remaining in the outflow of the SBBFR. The biofilm increased by 30-53% on the lava in the SBBFR that contained KTPPK after 11 days when compared with the reactor that contained P. putida KT2440. Real-time quantitative polymerase chain reaction confirmed that the copy of ppk was maintained at about 3.5 × 10(10) copies per μL general DNA in the biofilm after 20 days. Thus, the transgenic bacteria KTPPK could maintain a high density and promote phosphorus removal in the SBBFR. In summary, this study indicates that the use of SBBFR with transgenic bacteria has the potential to remove phosphorus and nitrogen from wastewater.     

Nutrient removal from slaughterhouse wastewater in an intermittently aerated sequencing batch reactor

The performance of a 10 L sequencing batch reactor (SBR) treating slaughterhouse wastewater was examined at ambient temperature. The influent wastewater comprised 4672+/-952 mg chemical oxygen demand (COD)/L, 356+/-46 mg total nitrogen (TN)/L and 29+/-10 mg total phosphorus (TP)/L. The duration of a complete cycle was 8 h and comprised four phases: fill (7 min), react (393 min), settle (30 min) and draw/idle (50 min). During the react phase, the reactor was intermittently aerated with an air supply of 0.8L/min four times at 50-min intervals, 50 min each time. At an influent organic loading rate of 1.2g COD/(Ld), average effluent concentrations of COD, TN and TP were 150 mg/L, 15 mg/L and 0.8 mg/L, respectively. This represented COD, TN and TP removals of 96%, 96% and 99%, respectively. Phase studies show that biological phosphorus uptake occurred in the first aeration period and nitrogen removal took place in the following reaction time by means of partial nitrification and denitrification. The nitrogen balance analysis indicates that denitrification and biomass synthesis contributed to 66% and 34% of TN removed, respectively.     

Effects of temperature, salinity, and carbon: nitrogen ratio on sequencing batch reactor treating shrimp aquaculture wastewater

In order to improve the water quality in the shrimp aquaculture, we tested a sequencing batch reactor (SBR) for the treatment of shrimp wastewater. A SBR is a variation of the activated sludge biological treatment process. This process uses multiple steps in the same tank to take the place of multiple tanks in a conventional treatment system. The SBR accomplishes pH correction, aeration, and clarification in a timed sequence, in a single reactor basin. This is achieved in a simple tank, through sequencing stages, which includes fill, react, settle, decant, and idle. The wastewater from the Waddell Mariculture Center, South Carolina was successfully treated using a SBR. The wastewater contained high concentration of carbon and nitrogen. By operating the reactor sequentially, viz, aerobic, anaerobic, and aerobic modes, nitrification and denitrification were achieved as well as removal of carbon. We optimized various environmental parameters such as temperature, salinity, and carbon and nitrogen ratio (C:N ratio) for the best performance of SBR. The results indicated that the salinity of 28-40 parts per thousand (ppt), temperature range of 22-37 degrees C, and a C:N ratio of 10:1 produced best results in terms of maximum nitrogen and carbon removal from the wastewater. The SBR system showed promising results and could be used as a viable treatment alternative in the shrimp industry.     

A pilot-scale study of struvite precipitation in a stirred tank reactor: conditions influencing the process

Currently, the two most developed techniques for recovering phosphorus from wastewater consist of the formation of calcium phosphates and struvite (MgNH(4)PO(4).6H(2)O). In this work the influence of the operational conditions on the struvite precipitation process (pH in the reactor, hydraulic retention time, and magnesium:phosphorus, nitrogen:phosphorus, and calcium:magnesium molar ratios) have been studied. Twenty-three experiments with artificial wastewater were performed in a stirred reactor. In order to obtain the pH value maintenance during the crystallization process, a fuzzy logic control has been developed. High phosphorus removal efficiencies were reliably achieved precipitating the struvite as easily dried crystals or as pellets made up of agglomerated crystals.     

Enhanced COD and nitrogen removals for the treatment of swine wastewater by combining submerged membrane bioreactor (MBR) and anaerobic upflow bed filter (AUBF) reactor

In order to enhance performances of organics removal and nitrification for the treatment of swine wastewater containing high concentration of organic solids and nitrogen than conventional biological nitrogen removal process, a submerged membrane bioreactor (MBR) was followed by an anaerobic upflow bed filter (AUBF) reactor in this research (AUBF–MBR process). The AUBF reactor is a hybrid reactor, which is the combination of an anoxic filter for denitrification and upflow anaerobic sludge blanket (UASB) for acid fermentation. In the AUBF–MBR process, it showed a considerable enhancement of the effluent quality in terms of COD removal and nitrification. The submerged MBR could maintain more than 14,000 mg VSS/L of the biomass concentration. Total nitrogen (T-N) removal efficiency represented 60% when internal recycle ratio was three times of flow-rate (Q), although the nitrification occurred completely. Although the volatile fatty acids produced in AUBF reactor can enhance denitrification rate, but the AUBF–MBR process showed reduction of overall removal efficiency of the nitrogen due to the reduction of carbon source by methane production in the AUBF reactor compared to that of theoretical nitrogen removal efficiency.

Long-term operation of the submerged MBR showed that the throughputs of the submerged MBR were respectively 74, 63, and 31 days at 10, 15, and 30 L/m² h (LMH) of permeate flux. Resistance to filtration by rejected solid is the primary cause of fouling, however the priority of cake resistance (R_c) and fouling resistance (R_f) with respect to filtration phenomenon was different according to the amount of permeate flux. The submerged MBR, here, achieved a steady-state flux of 15 LMH at 0.4 atm. of trans-membrane pressure (TMP) but the flux can be enhanced in the future because shear force by tangential flow will be greater when multi-layer sheets of membrane were used.     

Biological treatment of shrimp production wastewater

Over the last few decades, there has been an increase in consumer demand for shrimp, which has resulted in its worldwide aquaculture production. In the United States, the stringent enforcement of environmental regulations encourages shrimp farmers to develop new technologies, such as recirculating raceway systems. This is a zero-water exchange system capable of producing high-density shrimp yields. The system also produces wastewater characterized by high levels of ammonia, nitrate, nitrite, and organic carbon, which make waste management costs prohibitive. Shrimp farmers have a great need for a waste management method that is effective and economical. One such method is the sequencing batch reactor (SBR). A SBR is a variation of the activated sludge biological treatment process. This process uses multiple steps in the same reactor to take the place of multiple reactors in a conventional treatment system. The SBR accomplishes equalization, aeration, and clarification in a timed sequence in a single reactor system. This is achieved through reactor operation in sequences, which includes fill, react, settle, decant, and idle. A laboratory scale SBR was successfully operated using shrimp aquaculture wastewater. The wastewater contained high concentrations of carbon and nitrogen. By operating the reactors sequentially, namely, aerobic and anoxic modes, nitrification and denitrification were achieved as well as removal of carbon. Ammonia in the waste was nitrified within 4 days. The denitrification of nitrate was achieved by the anoxic process, and 100% removal of nitrate was observed within 15 days of reactor operation.     

Post-anoxic denitrification driven by PHA and glycogen within enhanced biological phosphorus removal

The objective of this research was to interrogate and develop a better understanding for a process to achieve post-anoxic denitrification without exogenous carbon augmentation within enhanced biological phosphorus removal (EBPR). Sequencing batch reactors fed real wastewater and seeded with mixed microbial consortia were operated under variable anaerobic-aerobic-anoxic and organic carbon loading conditions. The process consistently achieved phosphorus and nitrogen removal, while the observed specific denitrification rates were markedly higher than expected for post-anoxic systems operated without exogenous organic carbon addition. Investigations revealed that post-anoxic denitrification was predominantly driven by glycogen, an intracellular carbon storage polymer associated with EBPR; moreover, glycogen reserves can be significantly depleted post-anoxically without compromising EBPR. Success of the proposed process is predicated on providing sufficient organic acids in the influent wastewater, such that residual nitrate carried over from the post-anoxic period is reduced and polyhydroxyalkanoate (PHA) synthesis occurs.     

基于响应面法对一株好氧反硝化菌脱氮效能优化

【目的】水体富营养化是当今我国水环境面临的重大水域环境问题,氮素超标排放是主要的引发因素之一。好氧反硝化菌构建同步硝化反硝化工艺比传统脱氮工艺优势更大。获得高效的好氧反硝化菌株并通过生长因子优化使脱氮效率达到最高。【方法】经过序批式生物反应器(Sequencing batch reactor,SBR)的定向驯化,筛选获得高效好氧反硝化菌株,采用响应面法优化好氧反硝化过程影响总氮去除效率的关键因子(碳氮、溶解氧、pH、温度)。【结果】从运行稳定的SBR反应器中定向筛选高效好氧反硝化菌株Pseudomonas T13,采用响应面法对碳氮比、pH和溶解氧关键因子综合优化获得在18 h内最高硝酸盐去除率95%,总氮去除率90%。该菌株的高效反硝化效果的适宜温度范围为25?30 °C;最适pH为中性偏碱;适宜的COD/NO3?-N为4:1以上;最佳溶解氧浓度在2.5 mg/L。【结论】从长期稳定运行的SBR反应器中筛选获得一株高效好氧反硝化菌Pseudomonas T13,硝酸盐还原酶比例占脱氮酶基因的30%以上,通过运行条件优化获得硝氮去除率达到90%以上,对强化废水脱氮工艺具有良好应用价值。     

Batch biological treatment of nitrogen deficient synthetic wastewater using Azotobacter supplemented activated sludge

Biological treatment of nitrogen deficient wastewaters are usually accomplished by external addition of nitrogen sources to the wastewater which is an extra cost item. As an alternative for effective biological treatment of nitrogen deficient wastewaters, the nitrogen fixing bacterium, Azotobacter vinelandii, was used in activated sludge and also in pure culture. Total organic carbon (TOC) removal performances of Azotobacter-added and free activated sludge cultures were compared at different initial TN/TOC ratios. The rate and extent of TOC removal were comparable for all cultures when initial TN/TOC ratio was larger than 0.12; however, both the rate and extent of TOC removal from nitrogen deficient (TN/TOC<12%) synthetic wastewater were improved by using Azotobacter-added activated sludge as compared to the Azotobacter-free activated sludge culture. More than 90% TOC removal was obtained with pure Azotobacter or Azotobacter-added activated sludge culture from a nitrogen deficient synthetic wastewater.     

Assessing the feasibility of achieving biological nutrient removal from wastewater at an Irish food processing factory

In Ireland, wastewaters emanating from the food industry typically contain elevated levels of nitrogen and phosphorus before treatment. Two pilot scale studies were performed to determine the feasibility of achieving biological N and P removal on-site at a food ingredients plant. The wastewater treated by the pilot reactors was that which resulted from the day-to-day production in the full-scale food ingredients plant. Both reactors were of the anaerobic/anoxic/oxic (A/A/O) design, however the sizing of the zones was varied in this study. In the first pilot study, while treating a wastewater of the following strength: 1008 mg COD/l; 30.1 mg NH4-N/l and 26.7 mg P/l, removal efficiencies of 93%, 99% and 98% were obtained for COD, NH4-N and P, respectively. In the second study, while operating at reduced hydraulic retention times and lower recycle rates, the pilot plant treated a wastewater of the following strength: 1757 mg COD/l; 62 mg NH4-N/l and 57 mg P/l, with removal efficiencies of 94%, 97% and 75% obtained for COD, NH4-N and P, respectively. This work showed that biological nutrient removal could be successfully applied to treatment of food industry wastewaters.     

Sudden failure of biological nitrogen and carbon removal in the full-scale pre-denitrification process treating cokes wastewater

A full-scale pre-denitrification process treating cokes wastewater containing toxic compounds such as phenols, cyanides and thiocyanate has shown good performance in carbon and nitrogen removal. However, field operators have been having trouble with its instability without being able to identify the causes. To clarify the main cause of these sudden failures of the process, comprehensive studies were conducted on the pre-denitrification process using a lab-scale reactor system with real cokes wastewater. First, the shock loading effects of three major pollutants were investigated individually. As the loading amount of phenol increased to 600 mg/L, more COD, TOC and phenol itself were flowed into the aerobic reactor, but phenol itself did not inhibit nitrification and denitrification, owing to the effect of dilution and its rapid biodegradation. Higher loading of ammonia or thiocyanate slightly enhanced the removal efficiency of organic matter, but caused the final discharge concentration of total nitrogen to be above its legal limit of 60 mg-N/L. Meanwhile, continuous inflow of abnormal wastewater collected during unstable operation of the full-scale pre-denitrification process, caused a sudden failure of nitrogen removal in the lab-scale process, like the removal pattern of the full-scale one. This was discovered to be due to the lack of inorganic carbon in the aerobic reactor where autotrophic nitrification occurs.