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.     

Effect of anoxic/aerobic phase fraction on N2O emission in a sequencing batch reactor under low temperature

Laboratory scale anoxic/aerobic sequencing batch reactor (A/O SBR) was operated around 15 °C to evaluate the effect of anoxic/aerobic phase fraction (PF) on N₂O emission. The ammonia removal exhibited a decrease trend with the increase of PF, while the highest total nitrogen removal was achieved at PF = 0.5. Almost all the N₂O was emitted during the aerobic phase, despite of the PF value. However, the net emission of N₂O was affected by PF. Under the premise of completely aerobic nitrification, the lowest N₂O emission was achieved at PF = 0.5, with a N₂O-N conversion rate of 9.8%. At lower PF (PF = 0.2), N₂O emission was stimulated by residual nitrite caused by uncompleted denitrification during the anoxic phase. On the other hand, the exhaustion of the easily degradable carbon was the major cause for the high N₂O emission at higher PF (PF = 0.5). The N₂O emission increased with the decreasing temperature. The time-weighted N₂O emission quantity at 15 °C was 2.9 times higher than that at 25 °C.     

Assessment of a bioaugmentation strategy with polyphosphate accumulating organisms in a nitrification/denitrification sequencing batch reactor

Different alternative configurations and strategies for the simultaneous biological removal of organic matter and nutrients (N and P) in wastewater have been proposed in the literature. This work demonstrates a new successful strategy to bring in enhanced biological phosphorus removal (EBPR) to a conventional nitrification/denitrification system by means of bioaugmentation with an enriched culture of phosphorus accumulating organisms (PAO). This strategy was tested in a sequencing batch reactor (SBR), where an 8 h configuration with 3 h anoxic, 4.5 h aerobic and 25 min of settling confirmed that nitrification, denitrification and PAO activity could be maintained for a minimum of 60 days of operation after the bioaugmentation step. The successful bioaugmentation strategy opens new possibilities for retrofitting full-scale WWTP originally designed for only nitrification/denitrification. These systems could remove P simultaneously to COD and N if they were bioaugmented with waste purge of an anaerobic/aerobic SBR operated in parallel treating part of the influent wastewater.     

Enrichment of denitrifying glycogen-accumulating organisms in anaerobic/anoxic activated sludge system

Denitrifying glycogen-accumulating organisms (DGAO) were successfully enriched in a lab-scale sequencing batch reactor (SBR) running with anaerobic/anoxic cycles and acetate feeding during the anaerobic period. Acetate was completely taken up anaerobically, which was accompanied by the consumption of glycogen and the production of poly-beta-hydroxy-alkanoates (PHA). In the subsequent anoxic stage, nitrate or nitrite was utilized as electron acceptor for the oxidation of PHA, resulting in glycogen replenishment and cell growth. The above phenotype showed by the enrichment culture demonstrates the existence of DGAO. Further, it was found that the anaerobic behavior of DGAO could be predicted well by the anaerobic GAO model of Filipe et al. (2001) and Zeng et al. (2002a). The final product of denitrification during anoxic stage was mainly nitrous oxide (N(2)O) rather than N(2). The data strongly suggests that N(2)O production may be caused by the inhibition of nitrous oxide reductase by an elevated level of nitrite accumulated during denitrification. The existence of these organisms is a concern in biological nutrient removal systems that typically have an anaerobic/anoxic/aerobic reactor sequence since they are potential competitors to the polyphosphate-accumulating organisms.     

Enhanced nutrient removal in three types of step feeding process from municipal wastewater

An anoxic/oxic step feeding process was improved to enhance nutrient removal by reconfiguring the process into (1) anaerobic/anoxic/oxic step feeding process or (2) modified University of Capetown (UCT) step feeding process. Enhanced nitrogen and phosphorus removal and optimized organics utilization were obtained simultaneously in the modified UCT type with both internal and sludge recycle ratios of 75% as well as anaerobic/anoxic/oxic volume ratio of 1:3:6. Specifically, the UCT configuration and optimized operational conditions lead to the enrichment of denitrifying phosphorus removal microorganisms and achieved improved anaerobic P-release and anoxic P-uptake activities, which were beneficial to the denitrifying phosphorus removal activities and removal efficiencies. Due to high mixed liquor suspended solid and uneven distributed dissolved oxygen, 35% of total nitrogen was eliminated through simultaneous nitrification and denitrification process in aerobic zones. Moreover, 62 ± 6% of influent chemical oxygen demands was involved in the denitrification or phosphorus release processes.     

Nitrogen removal from wastewater with a low COD/N ratio at a low oxygen concentration

The goal of the study was to determine the effectiveness of nitrification and denitrification and the kinetics of ammonia removal from a mixture of wastewater and anaerobic sludge digester supernatant in an SBR at limited oxygen concentration. In addition, the COD removal efficiency and sludge production were assessed.In the SBR cycle alternating aerobic and anaerobic phases occurred; in the aeration phase the dissolved oxygen (DO) concentration was below 0.7 mg O₂/L. The low DO concentration did not inhibit ammonia oxidation-nitrification and the efficiency was ca. 96-98%. However, a relatively high COD concentration in the effluent was detected. The values of K_m and V_max, calculated from the Michaelis-Menten equation, were 43 mg N-NH₄/L and 15.64 mg N-NH₄/L h, respectively. Activated sludge production was almost stable (0.62-0.66 g MLVSS/g COD). A high net biomass production resulted from a low specific biomass decay rate of 0.0015 d⁻¹.     

Impact of COD/N ratio on nitrous oxide emission from microcosm wetlands and their performance in removing nitrogen from wastewater

Constructed wetlands (CWs) are considered to be important sources of nitrous oxide (N₂O). In order to investigate the effect of influent COD/N ratio on N₂O emission and control excess emission from nitrogen removal, free water surface microcosm wetlands were used and fed with different influent. In addition, the transformation of nitrogen was examined for better understanding of the mechanism of N₂O production under different operating COD/N ratios. It was found that N₂O emission and the performance of microcosm wetlands were significantly affected by COD/N ratio of wastewater influent. Strong relationships exist between N₂O production rate and nitrite (r = 0.421, p < 0.01). During denitrification process, DO concentration crucially influences N₂O production rate. An optimal influent COD/N ratio was obtained by adjusting external carbon sources for most effective N₂O emission control and best performance of the CWs in nitrogen removal from wastewater. It is concluded that under the operating condition of COD/N ratio = 5, total N₂O emission is minimum and the microcosm wetland is most effective in wastewater nitrogen removal.     

Temporal Change in Nitrous Oxide and Dinitrogen from Denitrification Following Onset of Anaerobiosis

Similar temporal patterns were found in three mineral soils for the composition of the gaseous products of denitrification following the onset of anaerobic conditions. During the early period of anaerobiosis (0 up to 1 to 3 h), N₂ was the dominant product of denitrification. The NO₃⁻ → N₂O activity then increased, but was not accompanied by a corresponding increase in N₂O-reducing activity. This resulted in a relatively extended period of time (1 to 3 up to 16 to 33 h) during which N₂O was a major product. Eventually (after 16 to 33 h), an increase in N₂O-reducing activity occurred without a comparable increase in the N₂O-producing activity. The increase in the rate of N₂O reduction did not occur in the presence of chloramphenicol and required the presence of N₂O or NO₃⁻ during the preceding anaerobic incubation. During the final period (16 to 33, up to 48 h), N₂ was generally the sole product of denitrification, since the rate of N₂O reduction exceeded the rate of N₂O production. A similar sequential pattern was also found for a culture of a denitrifying Flavobacterium sp. shifted to anaerobic growth. A staggered synthesis of the enzymes in the denitrification sequence apparently occurred in response to anoxia, which caused first a net production of N₂O followed by consumption of N₂O.     

Short term effect of ploughing a permanent pasture on N2O production from nitrification and denitrification

Soils are an important source of N₂O, which can be produced both in the nitrification and the denitrification processes. Grassland soils in particular have a high potential for mineralization and subsequent nitrification and denitrification. When ploughing long term grassland soils, the resulting high supply of mineral N may provide a high potential for N₂O losses. In this work, the short-term effect of ploughing a permanent grassland soil on gaseous N production was studied at different soil depths. Fertiliser and irrigation were applied in order to observe the effect of ploughing under a range of conditions. The relative proportions of N₂O produced from nitrification and denitrification and the proportion of N₂ gas produced from denitrification were determined using the methyl fluoride and acetylene specific inhibitors. Irrespectively to ploughing, fertiliser application increased the rates of N₂O production, N₂O production from nitrification, N₂O production from denitrification and total denitrification (N₂O + N₂). Application of fertiliser also increased the denitrification N₂O/N₂ ratio both in the denitrification potential and in the gaseous N productions by denitrification. Ploughing promoted soil organic N mineralization which led to an increase in the rates of N₂O production, N₂O production from nitrification, N₂O production from denitrification and total denitrification (N₂O + N₂). In both the ploughed and unploughed treatments the 0–10 cm soil layer was the major contributing layer to gaseous N production by all the above processes. However, the contribution of this layer decreased by ploughing, gaseous N productions from the 10 to 30 cm layer being significantly increased with respect to the unploughed treatment. Ploughing promoted both nitrification and denitrification derived N₂O production, although a higher proportion of N₂O lost by denitrification was observed as WFPS increased. Recently ploughed plots showed lower denitrification derived N₂O percentages than those ploughed before as a result of the lower soil water content in the former plots. Similarly, a lower mean nitrification derived N₂O percentage was found in the 10–30 cm layer compared with the 0–10 cm.     

Identifying sources of nitrous oxide emission in anoxic/aerobic sequencing batch reactors (A/O SBRs) acclimated in different aeration rates

Both long term and batch experiments were carried out to identify the sources of the N₂O emission in anoxic/aerobic sequencing batch reactors (A/O SBRs) under different aeration rates. The obtained results showed that aeration rate has an important effect on the N₂O emission of A/O SBR and most of the N₂O was emitted during the aerobic phase. During the anoxic phase, nitrate ammonification was the major source of N₂O emission while denitrification performed as a sink of N₂O, in all three bioreactors. The N₂O emission mechanisms during the aerobic phase differed with the aeration rate. At low and high aeration rates (Run 1 and Run 3), both coupled-denitrification and nitrifier denitrification were ascribed to be the source of N₂O emission. At mild aeration rate (Run 2), nitrifier denitrification by Nitrosomonas-like ammonia oxidizing-bacterial (AOB) was responsible for N₂O emission while coupled-denitrification turned out to be a sink of N₂O because of the presence of inner anaerobic region in sludge flocs.     

Characteristics of nitrous oxide (N2O) emission from intermittently aerated sequencing batch reactors (IASBRs) treating slaughterhouse wastewater at low temperature

This study investigated the characteristics of nitrous oxide (N₂O) emission from intermittently aerated sequencing batch reactors (IASBRs) treating high strength slaughterhouse wastewater at 11 °C, where partial nitrification followed by denitrification (PND) was achieved. N₂O generation and emission was examined at three aeration rates of 0.4, 0.6, and 0.8 L air/min in three IASBRs (SBR1, SBR2, and SBR3, respectively). The slaughterhouse wastewater contained chemical oxygen demand (COD) of 6057 ± 172.6 mg/L, total nitrogen (TN) of 576 ± 15.1 mg/L, total phosphorus (TP) of 52 ± 2.7 mg/L and suspended solids (SS) of 1843 ± 280.5 g/L. In the pseudo-steady state, the amount of N₂O emission was up to 5.7–11.0% of incoming TN. The aeration rate negatively affected N₂O emission and the ratio of N₂O emission to incoming TN was reduced by 48.2% when the aeration rate was increased from 0.4 to 0.8 L air/min. Results showed that more N₂O was generated in non-aeration periods than in aeration periods. Lower DO concentrations enhanced N₂O generation in the aeration periods (probably via nitrifier denitrification) while low DO concentrations (lower than 0.2 mg/L) did not affect N₂O generation in the non-aeration periods (probably via heterotrophic denitrification). When PHB was utilized as the organic substrate for denitrification, there was a high N₂O generation potential. It was estimated that 1.8 mg N₂O-N was generated accompanying per mg PHB consumed.     

Simultaneous oxidation of ammonium and p-cresol linked to nitrite reduction by denitrifying sludge

The metabolic capability of denitrifying sludge to oxidize ammonium and p-cresol was evaluated in batch cultures. Ammonium oxidation was studied in presence of nitrite and/or p-cresol by 55 h. At 50 mg/L NH₄⁺-N and 76 mg/L NO₂⁻-N, the substrates were consumed at 100% and 95%, respectively, being N₂ the product. At 50 mg/L NH₄⁺-N and 133 mg/L NO₂⁻-N, the consumption efficiencies decreased to 96% and 70%, respectively. The increase in nitrite concentration affected the ammonium oxidation rate. Nonetheless, the N₂ production rate did not change. In organotrophic denitrification, the p-cresol oxidation rate was slower than ammonium oxidation. In litho-organotrophic cultures, the p-cresol and ammonium oxidation rates were affected at 133 mg/L NO₂⁻-N. Nonetheless, at 76 mg/L NO₂⁻-N the denitrifying sludge oxidized ammonium and p-cresol, but at different rate. Finally, this is the first work reporting the simultaneous oxidation of ammonium and p-cresol with the production of N₂ from denitrifying sludge.     

传统厌氧/好氧生物除磷与厌氧/缺氧反硝化除磷效能的比较

采用序批式反应器(SBR),对比厌氧/好氧(A/O)和厌氧/缺氧(A/A)2种运行模式对模拟生活和工业混合污水同时脱氮除磷的效能。结果表明:反硝化聚磷菌完全可以在厌氧/缺氧交替运行条件下得到富集,稳定运行的2种模式对有机物和P的去除率分别保持在90%和85%以上,且A/A SBR具有更强的释磷能力,其释磷量比A/O SBR高出1.2倍。进一步试验表明:磷的释放在有无硝酸盐的情况下效果是不同的。2个系统内污泥均有反硝化除磷能力,A/A SBR中所含反硝化聚磷菌(DPAO)的比例是A/O SBR的4.56倍。2种模式出水水质都能取得较好的效果,且能实现同步除磷脱氮,而反硝化除磷在生物除磷方面更具优势。     

Potential of Aerobic Denitrification by Pseudomonas stutzeri TR2 To Reduce Nitrous Oxide Emissions from Wastewater Treatment Plants

In contrast to most denitrifiers studied so far, Pseudomonas stutzeri TR2 produces low levels of nitrous oxide (N₂O) even under aerobic conditions. We compared the denitrification activity of strain TR2 with those of various denitrifiers in an artificial medium that was derived from piggery wastewater. Strain TR2 exhibited strong denitrification activity and produced little N₂O under all conditions tested. Its growth rate under denitrifying conditions was near comparable to that under aerobic conditions, showing a sharp contrast to the lower growth rates of other denitrifiers under denitrifying conditions. Strain TR2 was tolerant to toxic nitrite, even utilizing it as a good denitrification substrate. When both nitrite and N₂O were present, strain TR2 reduced N₂O in preference to nitrite as the denitrification substrate. This bacterial strain was readily able to adapt to denitrifying conditions by expressing the denitrification genes for cytochrome cd₁ nitrite reductase (NiR) (nirS) and nitrous oxide reductase (NoS) (nosZ). Interestingly, nosZ was constitutively expressed even under nondenitrifying, aerobic conditions, consistent with our finding that strain TR2 preferred N₂O to nitrite. These properties of strain TR2 concerning denitrification are in sharp contrast to those of well-characterized denitrifiers. These results demonstrate that some bacterial species, such as strain TR2, have adopted a strategy for survival by preferring denitrification to oxygen respiration. The bacterium was also shown to contain the potential to reduce N₂O emissions when applied to sewage disposal fields.Wastewater treatment processes produce one of the major greenhouse effect gases, nitrous oxide (N₂O) (7, 25, 30). The global warming potential of N₂O relative to that of carbon dioxide (CO₂) is 298 for a 100-year time horizon, and its concentration in the atmosphere continues to increase by about 0.26% per year (9). Nitrogen removal in wastewater treatment plants is essentially based on the activities of nitrifying and denitrifying microorganisms, both of which are inhabitants of activated sludge. Nitrifying bacteria aerobically oxidize ammonium to nitrite (NO₂⁻) and nitrate (NO₃⁻), which are then reduced anaerobically by denitrifying bacteria to gaseous nitrogen forms, such as N₂O and dinitrogen (N₂). It has long been known that N₂O can be produced during both nitrification and denitrification processes of wastewater treatment (3, 19, 23), but the cause of N₂O emission during the nitrification process was not clear. We recently showed, however, using activated sludge grown under conditions that mimicked a piggery wastewater disposal, that N₂O emission during the nitrification process depends on denitrification by ammonia-oxidizing bacteria (Nitrosomonas) (18). On the other hand, it is believed that denitrifying bacteria produce N₂O as a by-product when anaerobiosis is insufficient during the denitrification process, because N₂O reductase is the enzyme that is most sensitive to oxygen (6). Piggery wastewater, in particular, contains a high concentration of ammonia, and N₂O emission tends to take place during the nitrogen removal process (5, 10). Experiments on the removal of ammonia and organic carbon by the aerobic denitrifier Pseudomonas stutzeri SU2 (24) and the heterotrophic nitrifier-aerobic denitrifier Alcaligenes faecalis no. 4 (16, 17) have been reported as examples of bioaugmentation in piggery wastewater treatment. Reduction of N₂O emissions from pig manure compost by addition of nitrite-oxidizing bacteria has also been reported (11). However, there have been no reports of methods for reducing N₂O emissions by bioaugmentation using aerobic denitrifying bacteria.Takaya et al. isolated the aerobic denitrifying bacterium Pseudomonas stutzeri TR2 (26). The denitrification activity of strain TR2 was monitored in batch and continuous cultures, using denitrification and artificial wastewater media, and the strain was found to keep a distinct activity (producing N₂ from NO₃⁻) and to produce a very low level of N₂O at a dissolved oxygen (O₂) concentration of 1.25 mg liter⁻¹. Therefore, strain TR2 should be useful in the future for reducing N₂O emissions from wastewater treatment plants by bioaugmentation. To investigate the feasibility of using strain TR2 for future application to wastewater treatment processes, we examined its denitrification activity, N₂O production, growth rate, and expression of denitrifying genes in batch cultures, using a medium that mimics the composition found in nitrogen removal wastewater plants. Comparison of the properties of strain TR2 with those of well-characterized denitrifying bacteria revealed characteristics of the strain that favor denitrification, although it can also respire oxygen.     

An integrated metabolic model for the aerobic and denitrifying biological phosphorus removal

In this work, an integrated metabolic model for biological phosphorus removal is presented. Using a previously proposed mathematical model it was shown to be possible to describe the two known biological phosphorus removal processes, under aerobic and denitrifying conditions, with the same biochemical reactions, where only the difference in electron acceptor (oxygen and nitrate) is taken into account. Though, apart from the ATP/NADH ratio, the stoichiometry in those models is identical, different kinetic parameters were found. Therefore, a new kinetic structure is proposed that adequately describes phosphorus removal under denitrifying and aerobic conditions with the same kinetic equations and parameters. The ATP/NADH ratio (delta) is the only model parameter that is different for aerobic and denitrifying growth. With the new model, simulations of anaerobic/aerobic and anaerobic/denitrifying sequencing batch reactors (A(2) SBR and A/O SBR) were made for verification of the model. Not only short-term behavior, but also steady state, was simulated. The results showed very good agreement between model predictions and experimental results for a wide range of dynamic conditions and sludge retention times. Sensitivity analysis shows the influence of the model parameters and the feed substrate concentrations on both systems. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 434-450, 1997.     

Nitrogen removal in a wastewater treatment plant through biofilters: nitrous oxide emissions during nitrification and denitrification

In order to estimate N₂O emissions from immersed biofilters during nitrogen removal in tertiary treatments at urban wastewater treatment plants (WWTPs), a fixed culture from the WWTP of “Seine Centre” (Paris conurbation) was subjected to lab-scale batch experiments under various conditions of oxygenation and a gradient of methanol addition. The results show that during nitrification, N₂O emissions are positively related to oxygenation (R ² = 0.99). However, compared to the rates of ammonium oxidation, the percentage of emitted N₂O is greater when oxygenation is low (0.5–1 mgO₂ L⁻¹), representing up to 1% of the oxidized ammonium (0.4% on average). During denitrification, the N₂O emission reaches a significant peak when the quantity of methanol allows denitrification of between 66% and 88%. When methanol concentrations lead to a denitrification of close to 100%, the flows of N₂O are much lower and represent on average 0.2% of the reduced nitrate. By considering these results, we can estimate, the emissions of N₂O during nitrogen removal, at the “Seine Centre” WWTP, to approximately 38 kgN-N₂O day⁻¹.     

Effect of aspartate and glutamate on the fate of enhanced biological phosphorus removal process and microbial community structure

This study investigated the fate of enhanced biological phosphorus removal (EBPR) and changes in microbial speciation in a sequencing batch reactor (SBR) fed with aspartate and glutamate. It involved SBR operation for 288 days, batch tests for observation of metabolic functions together with microscopic and phylogenetic analyses. Polyphosphate accumulating organisms (PAOs) were observed in abundance with complete removal of phosphorus. Fluorescence in situ hybridization (FISH) combined with 4′,6-dia-midino-2-phenylindole (DAPI) staining confirmed the accumulation of polyphosphate by Rhodocyclus-related and Actinobacterial PAOs. Aspartate seemed to favor the competitive growth of Rhodocyclus-related PAOs since EBPR population used the common biochemical pathways followed by Rhodocyclus-related PAOs in the aspartate fed batch tests. In the glutamate fed batch reactors, however, Actinobacterial PAOs appeared to be competitively selected which explains the lower levels of PHA generation. Even though operational conditions did not change, effective EBPR could not be maintained during the latter part of the study.     

Impact of chlortetracycline on sequencing batch reactor performance for swine manure treatment

Treatment of aged (500 day, 4 °C stored) chlortetracycline (CTC; 0, 20, 40, 80 mg/L CTC)-amended swine manure using two cycle, 22 day stage anaerobic sequencing batch reactors (SBR) was assessed. Eighty milligrams per liter CTC treatment inhibited SBR treatment efficiencies, although total gas production was enhanced compared to the no-CTC treatment. The 20 and 40 mg/L CTC treatments resulted in either slight or no differences to SBR treatment efficiencies and microbial diversities compared to the no-CTC treatment, and were generally similar to no-CTC treatments upon completion of the first 22 day SBR cycle. All CTC treatments enhanced SBR gas generation, however CH₄ yields were lowest for the 80 mg/L CTC treatment (0.111 L CH₄/g tCOD) upon completion of the second SBR react cycle. After a 22 day acclimation period, the 80 mg/L CTC treatment inhibited methanogenesis due to acetate accumulation, and decreased microbial diversity and CH₄ yield compared to the no-CTC treatment.     

Production of medium-chain-length polyhydroxyalkanoates by activated sludge enriched under periodic feeding with nonanoic acid

The potential use of activated sludge for the production of medium-chain-length polyhydroxyalkanoates (MCL-PHAs) was investigated. The enrichment of bacterial populations capable of producing MCL-PHAs was achieved by periodic feeding with nonanoic acid in a sequencing batch reactor (SBR). Denaturing gradient gel electrophoresis analysis revealed Pseudomonas aeruginosa strains to be predominant in the bacterial community during the SBR process. The composition of PHA synthesized by the enriched biomass from nonanoic acid consisted of a large concentration (>89 mol%) of MCL monomer units and a small amount of short-chain-length monomer units. Under fed-batch fermentation with continuous feeding of nonanoic acid at a flow rate of 0.225 g/L/h and a C/N ratio of 40, a maximum PHA content of 48.6% dry cell weight and a conversion yield (Y_p/s) of 0.94 g/g were achieved. These results indicate that MCL-PHA production by activated sludge is a promising alternative to typical pure culture approaches.     

Impacts of grazing intensity on denitrification and N<Subscript>2</Subscript>O production in a semi-arid grassland ecosystem

N₂O production from denitrification in soils contributes to the enhanced greenhouse effect and the destruction of the stratospheric ozone. Ungulate grazing affects denitrification and the production of N₂O. The short-term effect of grazing on denitrification and N₂O production has been examined in several grassland ecosystems. However, the effects of long-term grazing have rarely been studied. We measured denitrification and N₂O production during the 2005 and 2006 growing seasons in a long-term (17 years) experiment that had five grazing intensities (GI; 0.00, 1.33, 2.67, 4.00 and 5.33 sheep ha⁻¹). We found that denitrification and N₂O production rates were seasonally variable during the measurement period, with higher values observed in summer and lower values found in spring and autumn. The grazed treatments resulted in decreased denitrification and N₂O production, primarily due to the reduced soil nitrate concentration and organic N content under the long-term grazing. This supported our hypothesis that long-term over-grazing suppresses denitrification and N₂O production. Although significant differences in denitrification and N₂O production were not found between the four GI, there was a general trend that cumulative denitrification and N₂O production decreased as grazing intensity increased, especially in 2006. Lower N losses via denitrification and N₂O production in the grazed plots, to some extent, may contribute to the mitigation of greenhouse gas emission and help to preserve soil N and ameliorate the negative impacts of grazing on plant growth, productivity, and ecological restoration processes in the temperate steppe in northern China.