Improved nitrogen removal in upflow anaerobic sludge blanket (UASB) reactors by incorporation of Anammox bacteria into the granular sludge.

Upflow anaerobic sludge blanket reactors may offer a number of advantages over conventional mixed-tank, SBR, and biofilm reactors, including high space-loading, low footprint, and resistance to shocks and toxins. In this study, we assessed the use of upflow anaerobic sludge blanket (UASB) reactor technology as applied to anaerobic ammonia removal, or Anammox. Four 200 ml UASB reactors were inoculated with 50% (by volume) anaerobic granular sludge and 50% flocular sludge from different sources (all with the potential for containing Anammox organisms). Tools used to assess the reactors included basic analyses, fluorescent in-situ hybridisation, and mathematical modelling, with statistical non-linear parameter estimation. Two of the reactors showed statistically identical Anammox activity (i.e., identical kinetic parameters), with good ammonia and nitrite removal (0.14 kgNHx m(-3) reactor day(-1), with 99% ammonia removal). The third reactor also demonstrated significant Anammox activity, but with poor identifiability of parameters. The fourth reactor had no statistical Anammox activity. Modelling indicated that poor identifiability and performance in the third and fourth reactors were related to an excess of reduced carbon, probably originating in the inoculum. Accumulation of Anammox organisms was confirmed both by a volume loading much lower than the growth rate, and response to a probe specific for organisms previously reported to mediate Anammox processes. Overall, the UASB reactors were effective as Anammox systems, and identifiability of the systems was good, and repeatable (even compared to a previous study in a rotating biological contactor). This indicates that operation, design, and analysis of Anammox UASB reactors specifically, and Anammox systems in general, are reliable and portable, and that UASB systems are an appropriate technology for this process.     

Impact of high saline wastewaters on anaerobic granular sludge functionalities.

Three UASB reactors were operated at different salinity levels in order to assess the effects on the granular sludge properties. High levels of activity inhibition were observed at sodium concentrations over 7 g Na(+)/L, which resulted in low applicable organic loading rates and VFA accumulation in reactor effluents. However, either sludge adaptation or selection for saline resistant microorganisms occurred, which could be concluded from the observed increase in the 50% activity inhibitory concentrations of sodium during continuous flow experiments. Changes in Na(+) susceptibility in time are likely to be expected when treating saline wastewaters. The latter was evidenced by the high sodium tolerance of granular methanogenic sludge coming from a full-scale industrial reactor treating such wastewater. High salinity conditions resulted in a reduced granule strength, predicting process instabilities during long term reactor operation. Batch tests showed that high sodium concentrations seemed to displace the calcium from the granular sludge, a factor known to affect anaerobic granules formation.     

Study on performance of granular ANAMMOX process and characterization of the microbial community in sludge.

Anaerobic Ammonium Oxidation (ANAMMOX) is a novel biological nitrogen removal process, which is regarded as the most economical process at present. In this paper, two lab-scale UASB reactors, one of which was inoculated with the mixture of anaerobic sludge and aerobic sludge, the other with river sediments, were started up, using the inorganic synthetic water containing ammonium and nitrite as influent. After 421 days' and 356 days operation respectively, the ammonium removal efficiencies in two reactors reached 94% and 86% respectively, the total nitrogen volumetric loading rates were 2.5 and 1.6 kgN/m3 x d. ANAMMOX granules were obtained in both reactors; the color of most granules was brown, but some of them were red. Based on the observation and studies on the microstructure of the granules, three kinds of ANAMMOX granular sludge formation mechanisms were proposed: adhering biofilm and disintegrated granular core mechanism, adhering biofilm and inorganic core mechanism and the self-coherence mechanism. For phylogenetic characterization of anaerobic ammonium oxidizers, 16S rDNA approach was performed using Planctomycetales-specific PCR amplification. The dominant anammox bacteria occupied more than 90% of Planctomycetales-specific bacteria, and 27% of all bacteria in reactors. The dominant anammox bacteria distantly related to all currently reported candidate anammox genera. Functional gene of amoA was analyzed to investigate the 'aerobic' ammonium-oxidizing bacteria in beta-Proteobacteria. The 'aerobic' ammonium-oxidizing bacteria were more diverse than anammox bacteria, but most of them clustered in anoxic ammonium-oxidizing Nitrosomonas eutropha/europaea groups. The composition of 'aerobic' ammonium-oxidizing bacteria is only 2% of all of bacteria in reactors.     

好氧硝化污泥启动厌氧氨氧化试验研究

以好氧硝化污泥为培养污泥,采用经稀释的猪场废水启动厌氧氨氧化反应器,经过125 d的培养,根据ASBR反应器出水水样监测结果显示:ASBR反应器稳定运行后NH4+-N、NO2--N的去除率分别达到91.7%、92.0%,说明采用ASBR反应器,接种好氧硝化污泥可成功启动厌氧氨氧化反应器,验证了利用厌氧氨氧化工艺处理类似养殖废水的高氨氮废水的可能性.     

Rapid start-up of a nitrifying reactor using aerobic granular sludge as seed sludge

In this study, the effectiveness of aerobic granular sludge as seed sludge for rapid start-up of nitrifying processes was investigated using a laboratory-scale continuous stirred-tank reactor (CSTR) fed with completely inorganic wastewater which contained a high concentration of ammonia. Even when a large amount of granular biomass was inoculated in the reactor, and the characteristics of influent wastewater were abruptly changed, excess biomass washout was not observed, and biomass concentration was kept high at the start-up period due to high settling ability of the aerobic granular sludge. As a result, an ammonia removal rate immediately increased and reached more than 1.0 kg N/m(3)/d within 20 days and up to 1.8 kg N/m(3)/d on day 39. Subsequently, high rate nitritation was stably attained during 100 days. However, nitrite accumulation had been observed for 140 days before attaining complete nitrification to nitrate. Fluorescence in situ hybridization analysis revealed the increase in amount of ammonia-oxidizing bacteria which existed in the outer edge of the granular sludge during the start-up period. This microbial ecological change would make it possible to attain high rate ammonia removal.     

低负荷下厌氧氨氧化处理养殖废水的启动研究

厌氧氨氧化作为新型生物脱氮技术其关键在于如何实现厌氧氨氧化反应的启动,现有研究多以模拟废水为研究对象,本文以猪场废水为对象的研究,利用ASBR为反应器,接种反硝化污泥培养厌氧氨氧化细菌,在NH+4-N与NO-2-N浓度均为100 mg/L的条件下,运行125 d,经历启动初期、过渡期、系统稳定运行期三个阶段,厌氧氨氧化反应器中NH+4-N的去除率达91.70%,NO-2-N去除率92.0%;NH+4-N的容积负荷为36.90 mg/L.d,NO-2-N的容积负荷为37.55 mg/(L.d),成功实现了厌氧氨氧化反应器的启动。该研究成果对厌氧氨氧化技术在工程实践的应用具有重要的指导意义。     

EGSB反应器的启动运行研究

采用两个小试规模的EGSB(ExpandedGranularSludgeBed)反应器 ,分别接种厌氧絮状污泥和颗粒污泥来研究其启动规律。试验结果表明 ,接种厌氧絮状污泥的R1反应器 ,由于出水循环的采用导致严重的污泥流失 ,首先应按UASB反应器的运行方式培养出颗粒污泥后 ,再按EGSB反应器方式运行 ,共需 78d才能完成启动 ;接种厌氧颗粒污泥的R2反应器 ,采用适宜的回流比有利于细菌的生长和反应器运行效果的改善 ,在经过短暂的无回流驯化后 ,即可按EGSB运行方式启动运行 ,仅需 32d即完成启动。     

Comparing the performance of UASB and GRABBR treating low strength wastewaters

Anaerobic technologies have proved successful in the treatment of various high strength wastewaters with perceptible advantages over aerobic systems. The applicability of anaerobic processes to treat low strength wastewaters has been increasing with the evolution of high-rate reactors capable of achieving high sludge retention time (SRT) when operating at low HRT. However, the performance of these systems can be affected by high variations in flow and wastewater composition. This paper reports on the comparative study carried out with two such high rate reactors systems to evaluate their performances when used for the treatment of low strength wastewaters at high hydraulic rates. One of the two systems is the most commonly used upflow anaerobic sludge blanket (UASB) reactor in which all reactions occur within a single vessel. The other is the granular bed baffled reactor (GRABBR) that encourages different stages of anaerobic digestion in separate vessels longitudinally across the reactor. The reactors, with equal capacity of 10 litres, were subjected to increasing organic loading rates (OLRs) and hydraulic retention times (HRTs) of up to 60 kg COD m(-3) d(-1) and 1 h respectively. Results show that the GRABBR has greater processes stability at relatively low HRTs, whilst the UASB seems to be better equipped to cope with organic overloads or shockloads. The study also shows that the GRABBR enables the harvesting of biogas with greater energetic value and hence greater re-use potential than the UASB. Biogas of up to 86% methane content is obtainable with GRABBR treating low strength wastewaters.     

活性炭促进UASB处理高浓度啤酒废水效果研究

高浓度啤酒废水在厌氧消化过程中容易酸化,抑制甲烷转化。为提高UASB反应器的稳定性和处理效果,本研究通过序批式试验的方法,以模拟啤酒废水为底物,考察了两种不同粒径的活性炭对加速UASB污泥颗粒化进程和提高甲烷产率的影响。试验运行95 d,进水有机负荷从2.9 kg/(m3·d)增加到12.0 kg/(m3·d),监测反应器内产气量、出水VFA、出水TOC及污泥特性等参数的变化趋势。结果表明:投加活性炭能有效缩短厌氧污泥颗粒化的时间,增强产甲烷菌活性,大幅提升甲烷产量,出水TOC和VFA都维持在较低水平;并且较小粒径的粉末活性炭对UASB反应器的促进作用优于较大粒径的颗粒活性炭,能有效促进底物向甲烷气体转化。     

Enhanced granulation in UASB reactor treating low-strength wastewater by natural polymers.

Reetha (Sapindus trifoliata) seed extract and Chitosan were used as additives in the sludge bed of a UASB reactor treating low strength wastewater to enhance granulation. Five parallel laboratory scale UASB reactors were operated for 250 days with synthetic wastewater feed containing COD in the range of 600-800 mg/L. The reactors were seeded with spent sludge from a full-scale 5MLD UASB treatment plant at Jajmau, Kanpur, India. The seed sludge contained little or no granules. Different additives in the five reactors were as follows: control with no additive, cationic part of Reetha extract as additive, anionic part of Reetha extract as additive, bulk Reetha extract as additive and Chitosan as additive. The granulation rapidly increased in all the reactors beyond the 90th day of operation. The mean granule sizes as well as the fraction of granular sludge (particle size > or = 100 microm) were more in the presence of some of the additives compared to the control reactor. Chitosan significantly enhanced granulation followed by the cationic and anionic fractions of the Reetha extract. The bulk Reetha extract did not show enhancement of granulation. The ESEM/EDAX results showed that the bigger granules (3-4 mm) had porous structure and appeared as conglomerates of smaller granules.     

Stability and activity of anaerobic sludge from UASB reactors treating sewage in subtropical regions.

The production of small amounts of well-stabilized biological sludge is one of the main advantages of upflow anaerobic sludge bed (UASB) reactors over aerobic wastewater treatment systems. In this work, sludge produced in three pilot-scale UASB reactors used to treat sewage under subtropical conditions was assessed for both stability and specific methanogenic activity. Stability of primary sludge from settling tanks and digested sludge from conventional sludge digesters was also measured for comparison purposes. Kinetic parameters like the hydrolysis rate constant and the decay rate constant were calculated. High stability was observed in sludge from UASB reactors. Methanogenic activity in anaerobic sludges was relatively low, probably due to the low organic matter concentration in influent sewage. Knowledge on sludge growth rate, stability, and activity might be very useful to optimize sludge management activities in full-scale UASB reactors.     

Treatment of textile dyes in two-phase and single-phase anaerobic bio-treatment systems.

This research integrates two different concepts of anaerobic biotechnology- two-phase anaerobic treatment and anaerobic granular sludge bed technology, in treatment of colored wastewaters from textile industries. Four anaerobic reactors based on upflow anaerobic sludge blanket (UASB) technology were used as acid reactors and an expanded granular sludge bed (EGSB) reactor was used as a methane reactor. A conventional single-phase anaerobic reactor, working on EGSB technology was run in parallel to compare the performances of the two systems. Reactors were operated at different hydraulic retention times. The results from the study, which span over a period of 400 days, indicated that the two-phase system produces a higher quality of effluent in terms of color, COD and suspended solids than single-phase anaerobic treatment when operated under similar conditions. Alkalinity requirement of two-phase system was also observed to be lower than that of single-phase system which is important regarding design consideration.     

Nutrient removal and microbial granulation in an anaerobic process treating inorganic and organic nitrogenous wastewater.

The sustainable anaerobic nitrogen removal and microbial granulation were investigated by using a laboratory anaerobic granular sludge bed reactor, treating synthetic (inorganic and organic) wastewater and piggery waste. From inorganic synthetic wastewater, lithoautotrophic ammonium oxidation to nitrite/nitrate was observed by an addition of hydroxylamine. Also, the results revealed that the Anammox intermediates (particularly, hydrazine) contents in the substrate would be one of the important parameters for success of the anaerobic nitrogen removal process. The results from organic synthetic wastewater show that if the Anammox organism were not great enough in the startup of the process, denitritation and anaerobic ammonification would be a process prior to the Anammox reaction. The anaerobic ammonium removal from the piggery waste was performed successfully, probably due to the Anammox intermediates contained in the substrate. This reactor shows a complex performance including the Anammox reaction and HAP crystallization, as well as having partial denitritation occurring simultaneously. From the activity test, the maximum specific N conversion rate was 0.1 g NH4-N/g VSS/day (0.77 g T-N/g VSS/day), indicating that potential denitritation is quite high. The NO2-N/NH4-N ratio to Anammox is 1.17. The colour of the biomass treating the piggery waste changed from black to dark red. It was also observed that the red-colored granular sludge had a diameter of 1-2 mm. The settleability assessment of the granular sludge revealed that the granular sludge had a good settleability even though it was worse than that of seed granular sludge.     

Pretreatment of silk-dyeing industrial wastewater by UASB reactor.

The objective of this study was to investigate the performance of the upflow anaerobic sludge blanket (UASB) reactor as the pretreatment system for silk-dyeing wastewater. Two laboratory-scale UASB reactors, with working volume of 15.59 I, were used during May 1998 to June 1999. The actual wastewater was diluted to reduce ammonium ion toxicity on anaerobic bacteria. The experiments were conducted at the organic loading rates (OLRs) of 0.52, 1.01, 1.04, 1.54 and 2.56 kgCOD/(m3 x d), treating only wastewater generated from the acid-dye process of mixed-species raw silk. It took approximately 4 1/2 months to reach the steady-state conditions. It was found that the COD removal was in the ranges of 74.1-85.3%, except at OLR 2.56 kgCOD/(m3 x d) where efficiency significantly dropped to 55.2%. The apparent color removal was in the similar trend as COD. During the study periods, wastewater input had various color shades while the effluent generally looked pale yellowish. The methane generation rates ranged from 0.18-0.31 m3/kg COD removed, with methane composition 81.0-88.1% in biogas. The average granule size in the sludge bed had slowly increased to 0.73 mm in the last experiment. It can be concluded that the UASB reactor is suitable as a pretreatment system for silk-dyeing wastewater. An OLR of 1 kgCOD/(m3 x d) and an influent concentration diluted to 2,600 mgCOD/l are suggested while COD and apparent color removal efficiency of 80% and 70%, respectively, can be expected.     

Anammox process for synthetic and practical wastewater treatment using a novel kind of biomass carriers

The anammox process, as an alternative to conventional nitrogen removal technologies, has abstracted much attention in recent years. In this study, one column-type reactor using a novel support material--net type acrylic fiber (Biofix)--was used for anammox treatment. The Biofix reactor was operated at 25 degrees C (peak summer temperature, 31.5 degrees C). Over 330 days of operation for synthetic wastewater treatment, the nitrogen loading rates of the reactor were increased to 3.6 kg-N/m3/d and T-N removal efficiencies reached to 81.3%. For the practical anaerobic sludge digester liquor treatment, the average TN removal efficiency of 72% was obtained. A protein substance was shown to be the most abundant extracellular polymeric substances (EPS) in the granular sludge with almost two times more in the attached sludge of the Biofix reactor. Considering the EPS levels and observation by scanning electron microscopy, the anammox granules in the Biofix reactor were showing dense state. Results of DNA analyses indicated that the KSU-1 strain might prefer relatively low nutrient levels, while the KU2 strain might be better suited for the high media concentration. Other kinds of bacteria were also identified with the potentials for consuming the dissolved oxygen in the influent and facilitating anammox bacteria surviving under aerobic conditions.     

Investigation of the diversity of homoacetogenic bacteria in mesophilic and thermophilic anaerobic sludges using the formyltetrahydrofolate synthetase gene.

Homoacetogenic bacteria are strict anaerobes capable of autotrophic growth on H(2)/CO(2) or CO, and of heterotrophic growth on a wide range of sugars, alcohols, methoxylated aromatic compounds and one carbon compounds, yielding acetate as their sole metabolic end-product. Batch activity tests on anaerobic granular sludge, using H(2)/CO(2) as a substrate and 2-bromoethanesulfonate (BES) as a specific methanogenic inhibitor revealed that H(2)/CO(2) conversion and concomitant acetate production commenced only after a lag period of 60-100 h. This finding suggests that the homoacetogenic population of digester sludge could be maintained by heterotrophic growth on sugars or other organic compounds, rather than by autotrophic growth on H(2)/CO(2). In the present study, two upflow anaerobic sludge bed (UASB) reactors were operated at 37 degrees C and 55 degrees C for two distinct trial periods, each characterised by the application of influents designed to enrich for homoacetogenic bacteria. Specific primers designed for the amplification of the functional gene encoding formyltetrahydrofolate synthetase (FTHFS), a key enzyme in the acetyl-CoA pathway of acetogenesis, were used as a specific probe for acetogenic bacteria. The diversity of acetogens in the granular sludge cultivated in each reactor was revealed by application of FTHFS targeted PCR. Results show that biomass acetogenic composition was dependent upon the operational temperature of the reactor and the substrate supplied as influent.     

Difficulties in maintaining long-term partial nitritation of ammonium-rich sludge digester liquids in a moving-bed biofilm reactor (MBBR).

Nitrogen can be eliminated effectively from sludge digester effluents by anaerobic ammonium oxidation (anammox), but 55-60% of the ammonium must first be oxidized to nitrite. Although a continuous flow stirred tank reactor (CSTR) with suspended biomass could be used, its hydraulic dilution rate is limited to 0.8-1 d(-1) (30 degrees C). Higher specific nitrite production rates can be achieved by sludge retention, as shown here for a moving-bed biofilm reactor (MBBR) with Kaldnes carriers on laboratory and pilot scales. The maximum nitrite production rate amounted to 2.7 gNO2-Nm(-2)d(-1) (3 gO2m(-3)d(-1), 30.5 degrees C), thus doubling the dilution rate compared to CSTR operation with suspended biomass for a supernatant with 700 gNH4-Nm(-3). Whenever the available alkalinity was fully consumed, an optimal amount of nitrite was produced. However, a significant amount of nitrate was produced after 11 months of operation, making the effluent unsuitable for anaerobic ammonium oxidation. Because the sludge retention time (SRT) is relatively long in biofilm systems, slow growth of nitrite oxidizers occurs. None of the selection criteria applied - a high ammonium loading rate, high free ammonia or low oxygen concentration - led to selective suppression of nitrite oxidation. A CSTR or SBR with suspended biomass is consequently recommended for full-scale operation.     

Anaerobic ammonium oxidation of ammonium-rich waste streams in fixed-bed reactors.

The feasibility of anaerobic ammonium oxidation (Anammox) in fixed-bed reactors was evaluated on laboratory and pilot scales. Using synthetic wastewater, the specific nitrogen removal rate was increased from 0.05-0.1 kgNm(-3)(reactor)d(-1) to 0.35-0.38 kgNm(-3)(reactor)d(-1) within a year (T= 22-27 degrees C) in all applications. However, the anammox activity was seriously and repeatedly inhibited at prolonged high nitrite concentrations (e.g. six days at 30-50 gNO2-Nm(-3)) and recovery was always a lengthy process. But even at a moderate nitrite concentration (11+/-10 gNO2-Nm(-3)), the observed specific growth rate was only 0.018 d(-1) at 26.4+/-0.8 degrees C, which corresponds to approximately 0.025 d(-1) at 30 degrees C (doubling time: 28 days). In a second experimental period for another 250 days, one of the laboratory reactors was fed with partially nitrified sludge liquors from a domestic wastewater treatment plant (WWTP). In this case, the specific elimination rate was as high as 3.5 kgNm(-3)(reactor)d(-1) at 26-27 degrees C. Independently of the feed, the average nitrogen elimination rate lay between 80-85% in all applications. An appropriate hydraulic design is essential to prevent clogging and local nitrite inhibition in fixed-bed reactors.     

Grey water treatment in upflow anaerobic sludge blanket (UASB) reactor at different temperatures

The treatment of grey water in two upflow anaerobic sludge blanket (UASB) reactors, operated at different hydraulic retention times (HRTs) and temperatures, was investigated. The first reactor (UASB-A) was operated at ambient temperature (14-25 degrees C) and HRT of 20, 12 and 8 h, while the second reactor (UASB-30) was operated at controlled temperature of 30 degrees C and HRT of 16, 10 and 6 h. The two reactors were fed with grey water from 'Flintenbreite' settlement in Luebeck, Germany. When the grey water was treated in the UASB reactor at 30 degrees C, total chemical oxygen demand (CODt) removal of 52-64% was achieved at HRT between 6 and 16 h, while at lower temperature lower removal (31-41%) was obtained at HRT between 8 and 20 h. Total nitrogen and phosphorous removal in the UASB reactors were limited (22-36 and 10-24%, respectively) at all operational conditions. The results showed that at increasing temperature or decreasing HRT of the reactors, maximum specific methanogenic activity of the sludge in the reactors improved. As the UASB reactor showed a significantly higher COD removal (31-64%) than the septic tank (11-14%) even at low temperature, it is recommended to use UASB reactor instead of septic tank (the most common system) for grey water pre-treatment. Based on the achieved results and due to high peak flow factor, a HRT between 8 and 12 h can be considered the suitable HRT for the UASB reactor treating grey water at temperature 20-30 degrees C, while a HRT of 12-24 h can be applied at temperature lower than 20 degrees C.     

Anaerobic bioprocessing of sewage sludge, focusing on degradation of linear alkylbenzene sulfonates (LAS).

Anaerobic degradation of sludge amended with linear alkylbenzene sulfonates (LAS) was tested in a one stage continuous stirred tank reactor (CSTR) and a two stage reactor system consisting of a CSTR as first step and upflow anaerobic sludge bed (UASB) reactor as the second step. Anaerobic removal of LAS was only observed at the second step but not at the first step. Removal of LAS in the UASB reactors was approx. 80% where half was due to absorption and the other half was apparently due to biological removal as shown from the LAS mass balance. At the end of the experiment the reactors were spiked with 14C-LAS which resulted in 5.6% 14CO2 in the produced gas. Total mass balance of the radioactivity was however not achieved. In batch experiments it was found that LAS at concentrations higher than 50 mg/l is inhibitory for most microbial groups of the anaerobic process. Therefore, low initial LAS concentration is a prerequisite for successful LAS degradation. The results from the present study suggest that anaerobic degradation of LAS is possible in UASB reactors when the concentration of LAS is low enough to avoid inhibition of microorganisms active in the anaerobic process.