Simultaneous methanogenesis and denitrification of pretreated effluents from a fish canning industry

A lab-scale hybrid upflow sludge bed-filter (USBF) reactor was employed to carry out methanogenesis and denitrification of the effluent from an anaerobic industrial reactor (EAIR) in a fish canning industry. The reactor was initially inoculated with methanogenic sludge and there were two different operational steps. During the first step (Step I: days 1-61), the methanogenic process was carried out at organic loading rates (OLR) of 1.0-1.25 g COD l-1 d-1 reaching COD removal percentages of 80%. During the second step (Step II: days 62-109) nitrate was added as KNO3 to the industrial effluent and the OLR was varied between 1.0 and 1.25 g COD l-1 d-1. Two different nitrogen loads of 0.10 and 0.22 g NO3(-)-N l-1 d-1 were applied and these led to nitrogen removal percentages of around 100% in both cases and COD removal percentages of around 80%. Carbon to nitrogen ratio (C:N) in the influent was maintained at 2.0 and eventually it was increased to 3.0, by means of glucose addition, to control the denitrification process. From these results it is possible to establish that wastewater produced in a fish canning industry can be used as a carbon source for denitrification and that denitrifying microorganisms were present in the initially methanogenic sludge. Biomass productions of 0.23 and 0.61 g VSS:g TOC fed for Steps I and II, respectively, were calculated from carbon global balances, showing an increase in biomass growth due to denitrification.     

产甲烷UASB中颗粒污泥的快速培养及特性研究

在两相厌氧反应器的基础上,进行了产甲烷UASB中颗粒污泥的快速培养及特性研究.采用低负荷启动方式,通过快速提高进水COD和缩短水力停留时间使产甲烷UASB在最佳条件下运行,36d后产甲烷UASB中粒径>1.0 mm的颗粒污泥占64%,出水COD稳定在300 mg/L以下,可认为颗粒污泥培养成功.颗粒污泥的成熟经历了形成不规则核心、挤压架桥、分割成长、修剪长大四个阶段,加速污泥颗粒化的因素包括适宜的生长环境、良好的水力特征等.     

Development of a 2-sludge, 3-stage system for nitrogen and phosphorous removal from nutrient-rich wastewater using granular sludge and biofilms

A novel 2-sludge 3-stage process using a combination of granular sludge and biofilm was developed to achieve biological removal of nitrogen and phosphorus from nutrient-rich wastewater. The system consists of a granular sequencing batch reactor (SBR) working under alternating anaerobic/anoxic conditions supplemented with a short aerobic phase and an aerobic biofilm SBR. The wastewater is first fed to the granular SBR reactor, where easily biodegradable carbon sources are taken up primarily by polyphosphate accumulating organisms (PAOs). The supernatant resulting from quick settling of the granular sludge is then fed to the biofilm SBR for nitrification, which produces oxidized nitrogen that is returned to the granular reactor for simultaneous denitrification and phosphorus removal. While maximizing the utilization of organic substrates and reducing operational costs, as do other 2-sludge processes previously reported in literature, the proposed system solves the bottleneck problem of traditional 2-sludge systems, namely high effluent ammonia concentration, due to its high-volume exchange ratios. An ammonia oxidation rate of 32 mg N/Lh was achieved in the biofilm SBR, which produced nitrite as the final product. This nitrite stream was found to cause major inhibition on the anoxic P uptake and also to result in the accumulation of N(2)O. These problems were solved by feeding the nitrite-containing stream continuously to the granular reactor in the anoxic phase. With a nitrogen and phosphorus removal efficiency of 81% and 94%, respectively, the system produces an effluent that is suitable for land irrigation from a wastewater stream containing 270 mg N/L of total nitrogen and 40 mg P/L of total phosphorus.     

低浓度污水厌氧污泥颗粒化促进技术研究

向低浓度生活污水中投中絮凝剂（壳聚糖）和颗粒活性炭（GAC）的试验结果表明，这两种物质对厌氧污泥的颗粒化均有促进作用，但不如中、高浓度污水中明显。二者相比，GAC的效果更好一些（反应器的运行稳定，耐冲击能力强，出水水质良好），各反应器中形成的颗粒污泥均以产甲烷丝状菌为主。     

Effect of temperature on the anaerobic thermophilic conversion of volatile fatty acids by dispersed and granular sludge

The effect of temperature on the rate of volatile fatty acid (VFA) conversion by thermophilic methanogenic sludge, cultivated at 55°C, was studied using both batch activity tests and continuous flow experiments. The temperature dependence of acetate conversion in the range between 37–70°C could be described by an Arrhenius derived model when dispersed sludge with a low specific activity was used. For this sludge the optimum acetate conversion rate was found at 65°C. However, the maximum acetate utilization rate was not affected by temperature in the range between 50°C to 65°C when granular sludge with a high specific methanogenic activity was used. Crushing the granules led to a 2 to 3 fold increase in the maximum activity at 60–65°C, indicating that the conversion rate was very likely limited by the diffusion rate of acetate into the granules. Similar results were obtained with butyrate as the substrate. The temperature dependence of the crushed granules was similar to that of the less active dispersed sludge. In contrast, the thermophilic propionate oxidation rate was highest with the intact granular sludge while a similar temperature dependence was found for both the granular and dispersed sludges. The affinity for VFA increased with decreasing temperature. This phenomenon was most pronounced for the granular sludge. The thermophilic treatment of a VFA-mixture in a UASB reactor appeared to be only slightly affected by temperature when moderately low loading rates were applied, i.e. 20 kg COD·m⁻³·d⁻¹. However, temperature had a strong effect applying loading rates of 40–90 kg COD·m⁻³·d⁻¹ accompanied with high effluent VFA concentrations. The results reveal a high thermostability of the thermophilic wastewater treatment process in the range 45–60°C if “high-rate” reactors with a granular sludge bed are used.     

DEAMOX--new biological nitrogen removal process based on anaerobic ammonia oxidation coupled to sulphide-driven conversion of nitrate into nitrite

This paper reports about the successful laboratory testing of a new nitrogen removal process called DEAMOX (DEnitrifying AMmonium OXidation) for treatment of typical strong nitrogenous wastewater such as baker's yeast effluent. The concept of this process combines the recently discovered anammox (anaerobic ammonium oxidation) reaction with autotrophic denitrifying conditions using sulphide as an electron donor for the production of nitrite from nitrate within an anaerobic biofilm. To generate sulphide and ammonia, a Upflow Anaerobic Sludge Bed (UASB) reactor was used as a pre-treatment step. The UASB effluent was split and partially fed to a nitrifying reactor (to generate nitrate) and the remaining part was directly fed to the DEAMOX reactor where this stream was mixed with the nitrified effluent. Stable process performance and volumetric nitrogen loading rates of the DEAMOX reactor well above 1000 mgN/l/d with total nitrogen removal efficiencies of around 90% were obtained after long-term (410 days) optimisation of the process. Important prerequisites for this performance are appropriate influent ratios of the key species fed to the DEAMOX reactor, namely influent N-NO(x)/N-NH(4) ratios >1.2 (stoichiometry of the anammox reaction) and influent S-H(2)S/N-NO(3) ratios >0.57 mgS/mgN (stoichiometry of the sulphide-driven denitrification of nitrate to nitrite). The paper further describes some characteristics of the DEAMOX sludge as well as the preliminary results of its microbiological characterisation.     

Effect of shock and mixed nitrophenolic loadings on the performance of UASB reactors

The effect of nitrophenolic shock loads on the performance of three bench-scale upflow anaerobic sludge blanket (UASB) reactors was studied using synthetic wastewater. Reactors R1, R2 and R3 were fed with 30 mg/L concentration of 2-nitrophenol (2-NP), 4-nitrophenol (4-NP) and 2,4-dinitrophenol (2,4-DNP), respectively, along with methanol (COD = 2000 mg/ L), sodium nitrate (NO3(-)-N=200mg/L), and other nutrients. The reactors were in continuous operation for more than 2 years before the shock loading study was performed. Five nitrophenolic shock loadings of 45, 60, 75, 90 and 120mg/L d were administrated by increasing the influent nitrophenolic concentration to 45, 60, 75, 90 and 120mg/L, respectively, while keeping hydraulic retention time as 24h. The shocks were given continuously for a period of 4 days before switching back to normal nitrophenolic loading (30mg/Ld). The reactors were allowed to recover to normal performance level before administrating the next nitrophenolic shock load. The study showed that the nitrophenolic shock load of as high as 120 mg/L d did not affect the reactors performance irreversibly. After resuming the normal nitrophenolic loading, it took almost 3-18 days for the reactors to recover from the shock effect. The study was further extended to assess the maximum possible mixed nitrophenolic loading (2NP:4NP:2,4:DNP = 1:1:1) to which 2,4-DNP acclimated granular sludge containing reactor (R3) can be exposed without hampering the reactor (R3) performance irreversibly. The reactor was able to achieve pseudo-steady-state at a mixed nitrophenolic loading of 180 mg/L d with more than 90% removal of all the three nitrophenols, but failed at a mixed nitrophenolic loading of 225 mg/Ld.     

Long-term effects of operating temperature and sulphate addition on the methanogenic community structure of anaerobic hybrid reactors

The diversity, population dynamics, and activity profiles of methanogens in anaerobic granular sludges from two anaerobic hybrid reactors treating a molasses wastewater both mesophilically (37 degrees C) and thermophilically (55 degrees C) during a 1081 day trial were determined. The influent to one of the reactors was supplemented with sulphate, after an acclimation period of 112 days, to determine the effect of competition with sulphate-reducing bacteria on the methanogenic community structure. Sludge samples were removed from the reactors at intervals throughout the operational period and examined by amplified ribosomal DNA (rDNA) restriction analysis (ARDRA) and partial sequencing of 16S rRNA genes. In total, 18 operational taxonomic units (OTUs) were identified, 12 of which were sequenced. The methanogenic communities in both reactors changed during the operational period. The seed sludge and the reactor biomass sampled during mesophilic operation, both in the presence and absence of sulphate, was characterised by a predominance of Methanosaeta spp. Following temperature elevation, the dominant methanogenic sequences detected in the non-sulphate supplemented reactor were closely related to Methanocorpusculum parvum. By contrast, the dominant OTUs detected in the sulphate-supplemented reactor upon temperature increase were related to the hydrogen-utilising methanogen, Methanobacterium thermoautotrophicum. The observed methanogenic community structure in the reactors correlated with the operational performance of the reactors during the trial and with physiological measurements of the reactor biomass. Both reactors achieved chemical oxygen demand (COD) removal efficiencies of over 90% during mesophilic operation, with or without sulphate supplementation. During thermophilic operation, the presence of sulphate resulted in decreased reactor performance (effluent acetate concentrations of >3000 mg/l and biogas methane content of <25%). It was demonstrated that methanogenic conversion of acetate at 55 degrees C was extremely sensitive to inhibition by sulphide (50% inhibition at 8-17 mg/l unionised sulphide at pH 7.6-8.0), while the conversion of H(2)/CO(2) methanogenically was favoured. The combination of experiments carried out demonstrated the presence of specific methanogenic populations during periods of successful operational performance.     

Anoxic treatment of phenolic wastewater in sequencing batch reactor

Studies were conducted on the anoxic phenol removal using granular denitrifying sludge in sequencing batch reactor at different cycle lengths and influent phenol concentrations. Results showed that removal exceeded 80% up to an influent phenol concentration of 1050 mg/l at 6 h cycle length, which corresponded to 6.4 kg COD/m3/d. Beyond this, there was a steep decrease in phenol and COD removal efficiencies. This was accompanied by an increase in nitrite concentration in the effluent. On an average, 1 g nitrate-N was consumed per 3.4 g phenol COD removal. Fraction of COD available for sludge growth was calculated to be 11%.     

Simultaneous nitrogen and phosphate removal in aerobic granular sludge reactors operated at different temperatures

The main biological conversions taking place in two lab-scale aerobic granular sludge sequencing batch reactors were evaluated. Reactors were operated at different temperatures (20 and 30 °C) and accomplished simultaneous COD, nitrogen and phosphate removal. Nitrogen and phosphate conversions were linked to the microbial community structure as assessed by fluorescent in situ hybridization (FISH) analysis. Anoxic tests were performed to evaluate the contribution of anoxic phosphate uptake to the overall phosphate removal and to clarify the denitrification pathway. Complete nitrification/denitrification and phosphate removal were achieved in both systems. A considerable fraction of the phosphate removal was coupled to denitrification (denitrifying dephosphatation). From the results obtained in anoxic batch experiments dosing either nitrite or nitrate, denitrification was proposed to proceed mainly via the nitrate pathway. Denitrifying glycogen-accumulating organisms (DGAOs) were observed to be the main organisms responsible for the reduction of nitrate to nitrite. A significant fraction of the nitrite was further reduced to nitrogen gas while being used as electron acceptor by denitrifying polyphosphate-accumulating organisms (PAO clade II) for anoxic phosphate uptake.     

短程硝化/厌氧氨氧化联合工艺处理含氨废水的研究

在SBR中接种普通好氧活性污泥,通过控制运行条件来实现短程硝化,同时提高厌氧生物转盘系统中厌氧氨氧化的氮负荷,使之与SBR出水中NO2--N的积累量相匹配,并将二者组合形成短程硝化/厌氧氨氧化自养脱氮工艺.处理含氨废水的试验结果表明:在SBR的进水NH4+-N为150～250 mg/L、温度为(28±2)℃、pH值为7～8、DO<1 mg/L的条件下,可实现稳定的短程硝化,NO2--N积累率达85%以上,NH4+-N负荷达0.129 kgN/(kgVSS·d),AOB和NOB的数量之比为103:1.将短程硝化出水加入NH4+-N后作为厌氧氨氧化反应器的进水,在(40±1)℃下可以达到自养脱氮的目的,对NH4+-N、NO2--N和TN的去除率分别达86%、97%和90%以上,TN容积负荷为0.488 kgN/(m3·d).     

Impact of inocula and growth mode on the molecular microbial ecology of anaerobic ammonia oxidation (anammox) bioreactor communities

The composition of distinctly inoculated granular anammox and biofilm-based completely autotrophic nitrogen removal over nitrite (CANON) bioreactors was investigated from start-up through continuous long-term operation via denaturing gradient gel electrophoresis (DGGE) and sequencing. The granular anammox reactor was seeded with sludge from an operational anammox reactor in Strass, Austria. The CANON reactor was seeded with activated sludge from a local wastewater treatment plant in New York City. The principal anammox bacteria (AMX) shifted from members related to Kuenenia stuttgartiensis present in the initial inoculum to members related to Candidatus Brocadia fulgida during pre-enrichment (before this study) and to members related to Candidatus Brocadia sp. 40 (during this study) in the granular reactor. AMX related to C. Brocadia sp. 40 were also enriched from activated sludge in the CANON reactor. The estimated doubling times of AMX in the granular and CANON reactors were 5.3 and 8.9 days, respectively, which are lower than the value of 11 days, reported previously. Both the granular anammox and CANON reactors also fostered significant amounts of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The fractions of AMX and two groups of NOB were generally similar in the granular anammox and CANON reactors. However, the diversity and fractions of AOB in the two reactors was markedly different. Therefore, it is suggested that the composition of the feed and extant substrate concentrations in the reactor likely select for the microbial community composition more than the inocula and reactor configuration. Further, such selection is not equivalent for all resident communities.     

ABR中厌氧颗粒污泥的微生物学特性

了解厌氧折流板反应器(ABR)内厌氧颗粒污泥的微生态结构对于颗粒污泥的培养具有指导意义，为此对ABR各隔室中厌氧颗粒污泥的微生物组成进行扫描电镜观察，并测定了其在不同基质中的比产甲烷活性和辅酶F420的含量。结果表明，ABR各隔室颗粒污泥的微生物组成差异较大，1^#隔室颗粒污泥表面以产酸菌为主，内部以产甲烷杆菌为主，2^#、3^#隔室的颗粒污泥中没有明显的优势菌，菌群多样复杂，4^#隔室颗粒污泥中的优势菌是索氏甲烷菌；1^#隔室颗粒污泥利用葡萄糖、乙酸的产甲烷活性较低，利用丙酸的产甲烷活性最高，2^#、3^#、4^#隔室颗粒污泥利用葡萄糖、乙酸的产甲烷活性较高，利用丙酸的产甲烷活性较低。ABR中颗粒污泥的辅酶F420沿隔室逐渐升高，与产甲烷活性的变化一致，也就是说F420可以反映颗粒污泥的产甲烷活性。     

Integrated removal of nitrate and carbon in an upflow anaerobic sludge blanket (UASB) reactor: Operating performance

Denitrification and methanogenesis of a synthetic wastewater containing volatile fatty acids and nitrate were obtained in a single-stage process using an upflow anaerobic sludge blanket (UASB) reactor. The reactor was initially inoculated with methanogenic granular sludge and was gradually adapted to nitrate by increasing the nitrate concentration in the influent. Excess carbon not utilized for denitrification was converted to methane. During steady-state at a loading of 336 mg NO₃-N/l/d (24 mmol NO₃/l/d) and 6600 mg COD/l/d more than 99% removal of both nitrate and carbon was achieved. Batch experiments with biomass from the reactor showed that approximately 90% of the added nitrate was recovered as nitrogen gas indicating that true dentrification occurred. This was further verified from mass balances over the reactor. The granules changed appearance during the first 5 months of operation being fluffy and buoyant, probably reflecting changes in the microbial composition induced by the presence of nitrate. However, during the next two months more dense granules with good settling abilities gradually established in the system making this kind of combined process feasible in a UASB reactor. Characterization of the produced granules showed that while the mean diameter and density was comparable to granules from purely methanogenic systems, although the strength was lower.     

Improved brine recycling during nitrate removal using ion exchange

Ion exchange technology is currently the best for removing nitrate from drinking water. However, problems related to the disposal of spent brine from regeneration of exhausted resins must be overcome so that ion exchange can be applied more widely and economically, especially in small communities. For this purpose, a novel spent brine recycling system using combined biological denitrification and sulfate reduction processes was developed for more efficient reuse of brine. A granular activated carbon (GAC) adsorption column was introduced as an additional step to prevent contamination of resins by bio-polymers and dissolved organics present in the bio-reactor effluent. Two upflow sludge blanket reactors (USBRs) were operated in series for 166 days to provide denitrification and sulfate reduction. The denitrification reactor provided a nitrate removal efficiency of 96% at a nitrate-N loading rate of 5.4 g NO3(-)-N/l d. The sulfate reduction efficiency of the sulfate reduction reactor remained approximately 62% at a sulfate loading rate of 1.8 g SO4(2-)/l d. Five ion exchange columns containing A520E resins were repeatedly operated in up to 25 cycles of service and regeneration using five kinds of brine: one virgin 3% NaCl and four differently recycled spent brines. Throughput decreased remarkably when the biologically recycled brine was not treated with the GAC column, probably due to the presence of bio-polymers and dissolved organic compounds. The sulfate reduction reactor placed after the denitrification step increased the bicarbonate concentration, which could be used as a co-regenerant with chloride. The inclusion of the sulfate reduction reactor into the conventional brine recycling system allowed more efficient reuse of brine, resulting in both reduced salt consumption and brine discharge.     

序批式反应器的好氧颗粒污泥特性研究

对序批式反应器中好氧颗粒活性污泥的形成过程、处理性能和颗粒分布特性进行了研究。结果表明，不同操作条件下产生了结构形态不同的颗粒污泥，沉降时间是颗粒污泥形成的主要因素，有机负荷对颗粒污泥的结构有一定影响。颗粒污泥反应器对溶解性COD的去除率可达90％，对氨氮的去除率为24％。颗粒污泥在反应器中分布不均匀，反应器底部MLSS高达8g／L，SVI为34mL／g，随着反应器高度的增加则MLSS值降低、SVI值提高；此外较大的颗粒污泥聚集在反应器的底部，较小的颗粒污泥和絮体分布在反应器的中上部。     

Long-term impact of anaerobic reaction time on the performance and granular characteristics of granular denitrifying biological phosphorus removal systems

Removal of nitrogen and phosphorus (P) from wastewater is successfully and widely practiced in systems employing both granular sludge technology and enhanced biological P removal (EBPR) processes; however, the key parameter, anaerobic reaction time (AnRT), has not been thoroughly investigated. Successful EBPR is highly dependent on an appropriate AnRT, which induces carbon and polyphosphate metabolism by phosphorus accumulating organisms (PAOs). Therefore, the long-term impact of AnRT on denitrifying P removal performance and granular characteristics was investigated in three identical granular sludge sequencing batch reactors with AnRTs of 90 (R1), 120 (R2) and 150 min (R3). The microbial community structures and anaerobic stoichiometric parameters related to various AnRTs were monitored over time. Free nitrite acid (FNA) accumulation (e.g., 0.0008–0.0016 mg HNO₂–N/L) occurred frequently owing to incomplete denitrification in the adaptation period, especially in R3, which influenced the anaerobic/anoxic intracellular intermediate metabolites and activities of intracellular enzymes negatively, resulting in lower levels of poly-P and reduced activity of polyphosphate kinase. As a result, the Accumulibacter-PAOs population decreased from 51 ± 2.5% to 43 ± 2.1% when AnRT was extended from 90 to 150 min, leading to decreased denitrifying P removal performance. Additionally, frequent exposure of microorganisms to the FNA accumulation and anaerobic endogenous conditions in excess AnRT cases (e.g., 150 min) stimulated increased extracellular polymeric substances (EPS) production by microorganisms, resulting in enhanced granular formation and larger granules (size of 0.6–1.2 mm), but decreasing anaerobic PHA synthesis and glycogen hydrolysis. Phosphorus removal capacity was mediated to some extent by EPS adsorption in granular sludge systems that possessed more EPS, longer AnRT and relatively higher GAOs.     

Performance of UASB reactor treating leachate from acidogenic fermenter in the two-phase anaerobic digestion of food waste

This study was conducted to investigate the performance of the upflow anaerobic sludge blanket (UASB) reactor treating leachate from acidogenic fermenter in the two-phase anaerobic digestion of food waste. The chemical oxygen demand (COD) removal efficiency was consistently over 96% up to the loading rates of 15.8 g COD/l d. The methane production rate increased to 5.51/l d. Of all the COD removed, 92% was converted to methane and the remaining presumably to biomass. At loading rates over 18.7 g COD/l d, the COD removal efficiency decreased due to sludge flotation and washout in the reactor, which resulted from short HRT of less than 10.6 h. The residual propionate concentration was the highest among the volatile fatty acids (VFA) in the effluent. The specific methanogenic activity (SMA) analysis showed that the VFA-degrading activity of granule was the highest for butyrate, and the lowest for propionate. Typical granules were found to be mainly composed of microcolonies of Methanosaeta. The size distribution of sludge particles indicated that partially granulated sludge could maintain the original structure of granular sludge and continue to gain size in the UASB reactor treating leachate from acidogenic fermenter.     

DEGRADATION OF AROCLOR 1242 IN A SINGLE-STAGE COUPLED ANAEROBIC/AEROBIC BIOREACTOR

Degradation of Aroclor 1242 was studied in granular biofilm reactors with limited aeration. An aerobic biphenyl degrader, Rhodococcus sp. M5, was used to supplement a natural bacterial population present in a “bioaugmented” reactor, while the “non-bioaugmented” reactor only contained natural granular sludge. The bioaugmentation, however appeared to have no effect on the reactor performance. Aroclor measurements showed its disappearance in both reactors with only 16–19% of Aroclor recovered from the reactor biomass and effluent. Simultaneously, a chlorine balance indicated that dechlorination occurred at a specific rate of 1.43 mg PCB (g volatile suspended solids)⁻¹ d⁻¹, which was comparable to the observed rate of Aroclor disappearance. Intermediates detected in both reactors were biphenyl, benzoic acid, and mono-hydroxybiphenyls. This suggests that a near-complete mineralization of Aroclor can be achieved in a single-stage anaerobic/aerobic system due to a combination of reductive and oxidative degradation mechanisms.