Biological phosphorus removal from abattoir wastewater at very short sludge ages mediated by novel PAO clade Comamonadaceae.

Recent increases in global phosphorus costs, together with the need to remove phosphorus from wastewater to comply with water discharge regulations, make phosphorus recovery from wastewater economically and environmentally attractive. Biological phosphorus (Bio-P) removal process can effectively capture the phosphorus from wastewater and concentrate it in a form that is easily amendable for recovery in contrast to traditional (chemical) phosphorus removal processes. However, Bio-P removal processes have historically been operated at medium to long solids retention times (SRTs, 10-20 days typically), which inherently increases the energy consumption while reducing the recoverable carbon fraction and hence makes it incompatible with the drive towards energy self-sufficient wastewater treatment plants. In this study, a novel high-rate Bio-P removal process has been developed as an energy efficient alternative for phosphorus removal from wastewater through operation at an SRT of less than 4 days. The process was most effective at an SRT of 2-2.5 days, achieving >90% phosphate removal. Further reducing the SRT to 1.7 days resulted in a loss of Bio-P activity. 16S pyrotag sequencing showed the community changed considerably with changes in the SRT, but that Comamonadaceae was consistently abundant when the Bio-P activity was evident. FISH analysis combined with DAPI staining confirmed that bacterial cells of Comamonadaceae arranged in tetrads contained polyphosphate, identifying them as the key polyphosphate accumulating organisms at these low SRT conditions. Overall, this paper demonstrates a novel, high-rate phosphorus removal process that can be effectively integrated with short SRT, energy-efficient carbon removal and recovery processes.

Phototrophic bacteria for nutrient recovery from domestic wastewater.

The organics and nutrients in industrial and domestic wastewater are increasingly being regarded as a valuable resource for energy and nutrient recovery. Emerging concepts to redesign wastewater treatment as resource recovery systems include the use of different bacteria and algae to partition carbon and nutrients to the particulate phase through assimilation or bio-accumulation. This study evaluates the use of purple phototrophic bacteria (PPB) (also known as purple non-sulphur bacteria or PNSB) for such a biological concentration process through a series of batch tests. The key objectives are to (a) demonstrate consistent selection and enrichment of PPB using infrared light in a non-sterile medium, and (b) achieve effective partitioning of soluble organics, ammonium and phosphate into the PPB culture. PPB were successfully enriched from pre-settled domestic wastewater within 2-3 days and identified as members of the order Rhodobacterales. Under anaerobic conditions with infrared irradiation the enrichment culture was able to simultaneously remove COD (63 ± 5%), NH4-N (99.6%-0.12 ± 0.03 mgN L(-1)) and PO4-P (88%-0.8 ± 0.6 mgP L(-1)) from primary settled domestic wastewater in 24 h. In this experiment, acetate was added as an additional carbon source to demonstrate the maximal nitrogen and phosphorous elimination potential. Almost all the COD removed was assimilated into biomass rather than oxidised to CO2, with the total COD actually increasing during the batch experiments due to phototrophic synthesis. NH4-N and PO4-P were also assimilated by the biomass rather than removed through destructive oxidation or accumulation. The process offers the opportunity to concentrate organics and macronutrients from wastewater in one solids stream that can be anaerobically digested to generate energy and recover nutrients from the concentrated digestate. Technical challenges include the design of a continuous reactor system, as well as efficient delivery of electrons, either through light or chemical sources.

Operating aerobic wastewater treatment at very short sludge ages enables treatment and energy recovery through anaerobic sludge digestion.

Conventional abattoir wastewater treatment processes for carbon and nutrient removal are typically designed and operated with a long sludge retention time (SRT) of 10-20 days, with a relatively high energy demand and physical footprint. The process also generates a considerable amount of waste activated sludge that is not easily degradable due to the long SRT. In this study, an innovative high-rate sequencing batch reactor (SBR) based wastewater treatment process with short SRT and hydraulic retention time (HRT) is developed and characterised. The high-rate SBR process was shown to be most effective with SRT of 2-3 days and HRT of 0.5-1 day, achieving >80% reduction in chemical oxygen demand (COD) and phosphorus and approximately 55% nitrogen removal. A majority of carbon removal (70-80%) was achieved by biomass assimilation and/or accumulation, rather than oxidation. Anaerobic degradability of the sludge generated in the high-rate SBR process was strongly linked to SRT, with measured degradability extent being 85% (2 days SRT), 73% (3 days), and 63% (4 days), but it was not influenced by digestion temperature. However, the rate of degradation for 3 and 4 days SRT sludge was increased by 45% at thermophilic conditions compared to mesophilic conditions. Overall, the treatment process provides a very compact and energy efficient treatment option for highly degradable wastewaters such as meat and food processing, with a substantial space reduction by using smaller reactors and a considerable net energy output through the reduced aerobic oxidation and concurrent increased methane production potential through the efficient sludge digestion.

Evaluating a strategy for maintaining nitrifier activity during long-term starvation in a moving bed biofilm reactor (MBBR) treating reverse osmosis concentrate.

A two-stage moving bed biofilm reactor (MBBR) was applied at the Bundamba advanced water treatment plant (AWTP) (Queensland, Australia) to treat the reverse osmosis concentrate (ROC) for inorganic nutrient removal. One of the operational challenges for the system was to cope with the large fluctuations of the ROC flow. This study investigated the decay rates of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) and biofilm detachment in MBBR during starvation for up to one month. An intermittent aeration strategy of 15 min aeration every 6 h was applied. This study also evaluated the activity recovery of both AOB and NOB after normal operation was resumed. The results showed that the activity loss of AOB and NOB was relatively minor (<20%) within 10 days of starvation, which ensured relatively quick recovery of ammonium removal when normal operation resumed. In contrast, the AOB and NOB activity loss reached 60-80% when the starvation time was longer than 20 days, resulting in slower recovery of ammonium removal after starvation. Starvation for less than 20 days didn't result in an apparent biomass detachment from carriers.

A sequencing batch reactor system for high-level biological nitrogen and phosphorus removal from abattoir wastewater.

A sequencing batch reactor (SBR) system is demonstrated to biologically remove nitrogen, phosphorus and chemical oxygen demand (COD) to very low levels from abattoir wastewater. Each 6 h cycle contained three anoxic/anaerobic and aerobic sub-cycles with wastewater fed at the beginning of each anoxic/anaerobic period. The step-feed strategy was applied to avoid high-level build-up of nitrate or nitrite during nitrification, and therefore to facilitate the creation of anaerobic conditions required for biological phosphorus removal. A high degree removal of total phosphorus (>98%), total nitrogen (>97%) and total COD (>95%) was consistently and reliably achieved after a 3-month start-up period. The concentrations of total phosphate and inorganic nitrogen in the effluent were consistently lower than 0.2 mg P l(-1) and 8 mg N l(-1), respectively. Fluorescence in situ hybridization revealed that the sludge was enriched in Accumulibacter spp. (20-40%), a known polyphosphate accumulating organism, whereas the known glycogen accumulating organisms were almost absent. The SBR received two streams of abattoir wastewater, namely the effluent from a full-scale anaerobic pond (75%) and the effluent from a lab-scale high-rate pre-fermentor (25%), both receiving raw abattoir wastewater as feed. The pond effluent contained approximately 250 mg N l(-1) total nitrogen and 40 mg P l(-1) of total phosphorus, but relatively low levels of soluble COD (around 500 mg l(-1)). The high-rate lab-scale pre-fermentor, operated at 37 degrees C and with a sludge retention time of 1 day, proved to be a cheap and effective method for providing supplementary volatile fatty acids allowing for high-degree of biological nutrient removal from abattoir wastewater.

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.

Elucidation of metabolic pathways in glycogen-accumulating organisms with in vivo 13C nuclear magnetic resonance.

Glycogen-accumulating organisms (GAOs) are found in enhanced biological phosphorus removal systems where they compete with polyphosphate-accumulating organisms for external carbon substrates. (13)C nuclear magnetic resonance ((13)C-NMR) was used to elucidate the metabolic pathways operating in an enriched GAO culture dominated by two known GAOs (81.2%). The experiments consisted of adding (13)C-acetate (labelled on position 1 or 2) to the culture under anaerobic conditions, and operating the culture through a cycle consisting of an anaerobic, an aerobic and a further anaerobic phase. The carbon transformations over the cycle were monitored using in vivo(13)C-NMR. The two-carbon moieties in hydroxybutyrate and hydroxyvalerate were derived from acetate, while the propionyl precursor of hydroxyvalerate was primarily derived from glycogen, with only a small fraction originating from acetate. Comparison of the labelling patterns in hydroxyvalerate at the end of the first and the second anaerobic periods in pulse experiments with 2-(13)C-acetate showed that the Entner-Doudoroff (ED) pathway was used for the breakdown of glycogen. This conclusion was further supported by the labelling pattern on glycogen observed in the pulse experiments with 1-(13)C-acetate, which can only be explained by the operation of ED with recycling of pyruvate and glyceraldehyde 3-phosphate via gluconeogenesis. The activity of the ED pathway for glycogen degradation by GAOs is demonstrated here for the first time. In addition, the decarboxylating part of the tricarboxylic acid cycle was confirmed to operate also under anaerobic conditions.

Endogenous metabolism of Candidatus Accumulibacter phosphatis under various starvation conditions.

The endogenous processes of Candidatus Accumulibacter phosphatis (referred to as Accumulibacter), a known polyphosphate-accumulating organism (PAO) responsible for enhanced biological phosphorus removal systems (EBPR), were characterized during 8-day starvation under anaerobic, anoxic, aerobic and intermittent aerobic-anaerobic conditions. A lab-scale EBPR culture with Accumulibacter representing over 85% of the entire bacterial population as quantified with fluorescence in-situ hybridization was used in the study. Cell decay rates were found to be negligible under anaerobic and anoxic conditions and may be ignored in activated sludge models. The decay rate under aerobic conditions was determined to be 0.03/d at 22 degrees C, considerably lower than the values commonly used in activated sludge modeling. Polyphosphate and glycogen were utilized simultaneously under anaerobic and anoxic conditions for maintenance energy production, with glycogen being the primary energy source until the glycogen content reached very low levels. Glycogen was used by Accumulibacter as the primary source of energy for maintenance under aerobic conditions in the absence of polyhydroxyalkanoates. However, Accumulibacter did not seem to use polyphosphate for energy production during aerobic starvation, clearly contrasting the anaerobic and particularly the anoxic case. Intermittent aerobic-anaerobic storage resulted in not only negligible cell decay rate, but also slower rates of glycogen and polyphosphate utilization, and may therefore be an effective strategy for long-term storage of EBPR sludge.

Advances in enhanced biological phosphorus removal: from micro to macro scale.

The enhanced biological phosphorus removal (EBPR) process has been implemented in many wastewater treatment plants worldwide. While the EBPR process is indeed capable of efficient phosphorus (P) removal performance, disturbances and prolonged periods of insufficient P removal have been observed at full-scale plants on numerous occasions under conditions that are seemingly favourable for EBPR. Recent studies in this field have utilised a wide range of approaches to address this problem, from studying the microorganisms that are primarily responsible for or detrimental to this process, to determining their biochemical pathways and developing mathematical models that facilitate better prediction of process performance. The overall goal of each of these studies is to obtain a more detailed insight into how the EBPR process works, where the best way of achieving this objective is through linking together the information obtained using these different approaches. This review paper critically assesses the recent advances that have been achieved in this field, particularly relating to the areas of EBPR microbiology, biochemistry, process operation and process modelling. Potential areas for future research are also proposed. Although previous research in this field has undoubtedly improved our level of understanding, it is clear that much remains to be learned about the process, as many unanswered questions still remain. One of the challenges appears to be the integration of the existing and growing scientific knowledge base with the observations and applications in practice, which this paper hopes to partially achieve.

Engineered ecosystem for sustainable on-site wastewater treatment.

A pilot-scale engineered ecosystem (PSEE) operated for over 2 years in sub-tropical conditions, produced an effluent with COD (median 38 mg/L) and TSS (median 3mg/L) levels comparable to that required by the AS/NZS 1547:2000 Onsite Domestic Wastewater Management standard. Only partial nitrification was achieved as dissimilatory nitrate reduction to ammonia occurred; however the level of NH(4)-N was reduced by 75% and total inorganic nitrogen by 53%. Phosphorus was not removed by the system due to the lack of regular sludge removal. Mass balances around the system showed that bacteria removed 36% of the influent nitrogen and 76% of the influent COD. Algae and plants were shown to remove 5% of the influent nitrogen, and 6% of the influent phosphorus. Challenges in developing a sustainable on-site wastewater treatment system were largely met by minimising chemical, energy and labour inputs, eliminating the need for frequent sludge handling, and creating an effluent quality suitable for re-use in non-potable applications. However, the sludge removal from the system needs to be adequately managed to avoid excessive accumulation as this can cause a range of negative impacts.

Modeling the aerobic metabolism of polyphosphate-accumulating organisms enriched with propionate as a carbon source.

In enhanced biological phosphorus removal (EBPR) systems, polyphosphate-accumulating organisms (PAOs) are primarily responsible for removing phosphate from wastewater. Propionate is an abundant carbon substrate in many EBPR plants and has been suggested to provide PAOs an advantage over their carbon competitors--the glycogen-accumulating organisms (GAOs). The aerobic metabolism of PAOs enriched with a propionate carbon source is studied in this paper. A metabolic model is proposed and experimentally validated to characterize the aerobic biochemical transformations by PAOs. The model predicts very well the experimental data obtained from the enriched PAO culture through solid-, liquid-, and gas-phase analyses. This model may be combined with previously formulated metabolic models to better describe the biochemical activity of PAOs with acetate and propionate as the primary carbon sources. Furthermore, it can also facilitate the study of the effect of different carbon sources on PAO-GAO competition.

Obtaining highly enriched cultures of Candidatus Accumulibacter phosphates through alternating carbon sources.

Candidatus Accumulibacter Phosphatis is widely considered to be a polyphosphate accumulating organism (PAO) of prime importance in enhanced biological phosphorus removal (EBPR) systems. This organism has yet to be isolated, despite many attempts. Previous studies on the biochemical and physiological aspects of this organism, as well as its response to different EBPR operational conditions, have generally relied on the use of mixed culture enrichments. One frequent problem in obtaining highly enriched cultures of this organism is the proliferation of glycogen accumulating organisms (GAO) that can compete with PAOs for limited carbon sources under similar operational conditions. In this study, Candidatus Accumulibacter Phosphatis has been enriched in a lab-scale bioreactor to a level greater than 90% as quantified by fluorescence in situ hyrbridisation (FISH). This is the highest enrichment of this organism that has been reported thus far, and was obtained by alternating the sole carbon source in the feed between acetate and propionate every one to two sludge ages, and operating the bioreactor within a pH range of 7.0-8.0. Simultaneously, the presence of two known groups of GAOs was eliminated under these operational conditions. Excellent phosphorus removal performance and stability were maintained in this system, where the phosphorous concentration in the effluent was below 0.2 mg/L for more than 7 months. When a disturbance was introduced to this system by adding sludge from an enriched GAO culture, Candidatus Accumulibacter Phosphatis once again became highly enriched, while the GAOs were out-competed. This feeding strategy is recommended for future studies focused on describing the physiology and biochemistry of Accumulibacter, where a highly-enriched culture of this organism is of high importance.

Identifying causes for N2O accumulation in a lab-scale sequencing batch reactor performing simultaneous nitrification, denitrification and phosphorus removal.

The recently described process of simultaneous nitrification, denitrification and phosphorus removal (SNDPR) has a great potential to save capital and operating costs for wastewater treatment plants. However, the presence of glycogen-accumulating organisms (GAOs) and the accumulation of nitrous oxide (N(2)O) can severely compromise the advantages of this process. In this study, these two issues were investigated using a lab-scale sequencing batch reactor performing SNDPR over a 5-month period. The reactor was highly enriched in polyphosphate-accumulating organisms (PAOs) and GAOs representing around 70% of the total microbial community. PAOs were the dominant population at all times and their abundance increased, while GAOs population decreased over the study period. Anoxic batch tests demonstrated that GAOs rather than denitrifying PAOs were responsible for denitrification. N(2)O accumulated from denitrification and more than half of the nitrogen supplied in a reactor cycle was released into the atmosphere as N(2)O. After mixing SNDPR sludge with other denitrifying sludge, N(2)O present in the bulk liquid was reduced immediately if external carbon was added. We therefore suggest that the N(2)O accumulation observed in the SNDPR reactor is an artefact of the low microbial diversity facilitated by the use of synthetic wastewater with only a single carbon source.

Competition between polyphosphate and glycogen accumulating organisms in enhanced biological phosphorus removal systems with acetate and propionate as carbon sources.

Enhanced biological phosphorus removal (EBPR) is a widely used process for achieving phosphorus removal from wastewater. A potential reason for EBPR failure is the undesirable growth of glycogen accumulating organisms (GAOs), which can compete for carbon sources with the bacterial group responsible for phosphorus removal from wastewater: the polyphosphate accumulating organisms (PAOs). This study investigates the impact of carbon source on EBPR performance and the competition between PAOs and GAOs. Two sequencing batch reactors (SBRs) were operated during a 4-6 month period and fed with a media containing acetate or propionate, respectively, as the sole carbon source. It was found that the acetate fed SBR rarely achieved a high level of phosphorus removal, and that a large portion of the microbial community was comprised of "Candidatus Competibacter phosphatis", a known GAO. The propionate fed SBR, however, achieved stable phosphorus removal throughout the study, apart from one brief disturbance. The bacterial community of the propionate fed SBR was dominated by "Candidatus Accumulibacter phosphatis", a known PAO, and did not contain Competibacter. In a separate experiment, another SBR was seeded with a mixture of PAOs and a group of alphaproteobacterial GAOs, both enriched with propionate as the sole carbon source. Stable EBPR was achieved and the PAO population increased while the GAOs appeared to be out-competed. The results of this paper suggest that propionate may provide PAOs with a selective advantage over GAOs in the PAO-GAO competition, particularly through the minimisation of Competibacter. Propionate may be a more suitable substrate than acetate for enhancing phosphorus removal in EBPR systems.

The effect of pH on the competition between polyphosphate-accumulating organisms and glycogen-accumulating organisms.

In enhanced biological phosphorus removal (EBPR) processes, glycogen-accumulating organisms (GAOs) may compete with polyphosphate-accumulating organisms (PAOs) for the often-limited carbon substrates, potentially resulting in disturbances to phosphorus removal. A detailed investigation of the effect of pH on the competition between PAOs and GAOs is reported in this study. The results show that a high external pH ( approximately 8) provided PAOs with an advantage over GAOs in EBPR systems. The phosphorus removal performance improved due to a population shift favouring PAOs over GAOs, which was shown through both chemical and microbiological methods. Two lab-scale reactors fed with propionate as the carbon source were subjected to an increase in pH from 7 to 8. The phosphorus removal and PAO population (as measured by quantitative fluorescence in situ hybridisation analysis of "Candidatus Accumulibacter phosphatis") increased in each system, where the PAOs appeared to out-compete a group of Alphaproteobacteria GAOs. A considerable improvement in the P removal was also observed in an acetate fed reactor, where the GAO population (primarily "Candidatus Competibacter phosphatis") decreased substantially after a similar increase in the pH. The results from this study suggest that pH could be used as a control parameter to reduce the undesirable proliferation of GAOs and improve phosphorus removal in EBPR systems.

Comparison of acetate and propionate uptake by polyphosphate accumulating organisms and glycogen accumulating organisms.

Enhanced biological phosphorus removal (EBPR) performance is directly affected by the competition between polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigates the effects of carbon source on PAO and GAO metabolism. Enriched PAO and GAO cultures were tested with the two most commonly found volatile fatty acids (VFAs) in wastewater systems, acetate and propionate. Four sequencing batch reactors (SBRs) were operated under similar conditions and influent compositions with either acetate or propionate as the sole carbon source. The stimulus for selection of the PAO and GAO phenotypes was provided only through variation of the phosphorus concentration in the feed. The abundance of PAOs and GAOs was quantified using fluorescence in situ hybridisation (FISH). In the acetate fed PAO and GAO reactors, "Candidatus Accumulibacter phosphatis" (a known PAO) and "Candidatus Competibacter phosphatis" (a known GAO) were present in abundance. A novel GAO, likely belonging to the group of Alphaproteobacteria, was found to dominate the propionate fed GAO reactor. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to take up acetate and propionate. PAOs enriched with acetate as the sole carbon source were immediately able to take up propionate, likely at a similar rate as acetate. However, an enrichment of GAOs with acetate as the sole carbon source took up propionate at a much slower rate (only about 5% of the rate of acetate uptake on a COD basis) during a short-term switch in carbon source. A GAO enrichment with propionate as the sole carbon source took up acetate at a rate that was less than half of the propionate uptake rate on a COD basis. These results, along with literature reports showing that PAOs fed with propionate (also dominated by Accumulibacter) can immediately switch to acetate, suggesting that PAOs are more adaptable to changes in carbon source as compared to GAOs. This study suggests that the PAO and GAO competition could be influenced in favour of PAOs through the provision of propionate in the feed or even by regularly switching the dominant VFA species in the wastewater. Further study is necessary in order to provide greater support for these hypotheses.

Optimisation of poly-beta-hydroxyalkanoate analysis using gas chromatography for enhanced biological phosphorus removal systems.

Poly-beta-hydroxyalkanoate (PHA) is a polymer commonly used in carbon and energy storage for many different bacterial cells. Polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), store PHA anaerobically through metabolism of carbon substrates such as acetate and propionate. Although poly-beta-hydroxybutyrate (PHB) and poly-beta-hydroxyvalerate (PHV) are commonly quantified using a previously developed gas chromatography (GC) method, poly-beta-hydroxy-2-methylvalerate (PH2MV) is seldom quantified despite the fact that it has been shown to be a key PHA fraction produced when PAOs or GAOs metabolise propionate. This paper presents two GC-based methods modified for extraction and quantification of PHB, PHV and PH2MV from enhanced biological phosphorus removal (EBPR) systems. For the extraction of PHB and PHV from acetate fed PAO and GAO cultures, a 3% sulfuric acid concentration and a 2-20 h digestion time is recommended, while a 10% sulfuric acid solution digested for 20h is recommended for PHV and PH2MV analysis from propionate fed EBPR systems.

Anaerobic metabolism of propionate by polyphosphate-accumulating organisms in enhanced biological phosphorus removal systems.

Propionate, a carbon substrate abundant in many prefermenters, has been shown in several previous studies to be a more favorable substrate than acetate for enhanced biological phosphorus removal (EBPR). The anaerobic metabolism of propionate by polyphosphate accumulating organisms (PAOs) is studied in this paper. A metabolic model is proposed to characterize the anaerobic biochemical transformations of propionate uptake by PAOs. The model is demonstrated to predict very well the experimental data from a PAO culture enriched in a laboratory-scale reactor with propionate as the sole carbon source. Quantitative fluorescence in-situ hybridization (FISH) analysis shows that Candidatus Accumulibacter phosphatis, the only identified PAO to date, constitute 63% of the bacterial population in this culture. Unlike the anaerobic metabolism of acetate by PAOs, which induces mainly poly-beta-hydroxybutyrate (PHB) production, the major fractions of poly-beta-hydroxyalkanoate (PHA) produced with propionate as the carbon source are poly-beta-hydroxyvalerate (PHV) and poly-beta-hydroxy-2-methylvalerate (PH2MV). PHA formation correlates very well with a selective (or nonrandom) condensation of acetyl-CoA and propionyl-CoA molecules. The maximum specific propionate uptake rate by PAOs found in this study is 0.18 C-mol/C-mol-biomass . h, which is very similar to the maximum specific acetate uptake rate reported in literature. The energy required for transporting 1 carbon-mole of propionate across the PAO cell membrane is also determined to be similar to the transportation of 1 carbon-mole of acetate. Furthermore, the experimental results suggest that PAOs possess a similar preference toward acetate and propionate uptake on a carbon-mole basis.

Simultaneous nitrification, denitrification, and phosphorus removal in a lab-scale sequencing batch reactor.

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic-enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the energy and COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen (DO) concentration (0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification, and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHAs), accompanied by phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to <0.5 mg/L by the end of the cycle. Ammonia was also oxidized during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis showed that the final denitrification product was mainly nitrous oxide (N(2)O), not N(2). Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen-accumulating organisms (DGAOs), rather than denitrifying polyphosphate-accumulating organisms (DPAOs), were responsible for the denitrification activity.