Nitrogen removal from wastewater using simultaneous nitrate reduction and anaerobic ammonium oxidation in single reactor

The effects of C/N ratio and total organic carbon (TOC) loading on nitrogen removal through simultaneous nitrate reduction and anaerobic ammonium oxidation in a single reactor were examined. Granular sludge taken from a methane fermentation reactor was placed in an upflow reactor and supplied with synthetic wastewater containing nitrate at a C/N ratio of 1 to grow heterotrophic denitrifying bacteria. When nitrogen removal ratio reached 30%, anammox sludge attached to nonwoven-carrier was added into the same reactor and then ammonia was added to the synthetic wastewater. Nitrogen removal ratio was markedly increased to 80-94%. In this system, nitrogen removal ratio was affected by C/N ratio and TOC loading, not by the amount of granular sludge. A stable isotopic analysis using 15N-labeled nitrate showed that N2 gas was formed by anammox reaction.     

Treatment of liquid fraction separated from liquidized food waste in an upflow anaerobic sludge blanket reactor

Thermochemical liquidization as a pretreatment for anaerobic digestion of food waste was studied using a laboratory-scale upflow anaerobic sludge blanket (UASB) reactor for a period of 82 d. Model food waste (approximately 90 wt% moisture content) was thermochemically liquidized at 175 degrees C for 1 h. The liquidized food waste was separated into a solid phase (6-10 wt%) and a liquid phase (85-89 wt%). The diluted liquid phase was continuously treated by anaerobic digestion using a UASB reactor at 35 degrees C. The volumetric loading rate was increased stepwise to 6.4-7.2 g total organic carbon (TOC)/l-reactor/d. Methane production was found to be approximately 0.35-0.61 l/g-TOC removed. The range of TOC removal efficiencies was 67-69% at an influent TOC concentration of 10.1-11.1 g/l and a volumetric loading rate of 4.8-5.3 g-TOC/l-reactor/d. This treatment process using an UASB reactor could be suitable for resource recovery from food waste.     

基于高分子载体与活性炭对比处理白酒废水的研究

实验以高分子材料做载体,与多孔颗粒活性炭(GAC)在完全相同的条件下,作厌氧流化床固定化微生物处理白酒废水的对比性研究.6d后加入已经分离出来的高效厌氧微生物,结果表明,新型高分子载体能有效促进厌氧菌固定化,可比GAC缩短反应器启动时间12d左右;且在进水COD值为3957.4mg/L、pH值为7.30、膨胀率为50%、水力停留时间为9h的条件下,COD的云除率为90.5%.     

Acceleration of cellulose degradation and shift of product via methanogenic co-culture of a cellulolytic bacterium with a hydrogenotrophic methanogen

Although the effects of syntrophic relationships between bacteria and methanogens have been reported in some environments, those on cellulose decomposition using cellulolytic bacteria from methanogenic reactors have not yet been examined. The effects of syntrophic co-culture on the decomposition of a cellulosic material were investigated in a co-culture of Clostridium clariflavum strain CL-1 and the hydrogenotrophic methanogen Methanothermobacter thermautotrophicus strain ΔH and a single-culture of strain CL-1 under thermophilic conditions. In this study, strain CL-1 was newly isolated as a cellulolytic bacterium from a thermophilic methanogenic reactor used for degrading garbage slurry. The degradation efficiency and cell density of strain CL-1 were 2.9- and 2.7-fold higher in the co-culture than in the single-culture after 60?h of incubation, respectively. Acetate, lactate and ethanol were the primary products in both cultures, and the concentration of propionate was low. The content of acetate to total organic acids plus ethanol was 59.3% in the co-culture. However, the ratio decreased to 24.9% in the single-culture, although acetate was the primary product. Therefore, hydrogen scavenging by the hydrogenotrophic methanogen strain ΔH could shift the metabolic pathway to the acetate production pathway in the co-culture. Increases in the cell density and the consequent acceleration of cellulose degradation in the co-culture would be caused by increases in adenosine 5'-triphosphate (ATP) levels, as the acetate production pathway includes ATP generation. Syntrophic cellulose decomposition by the cellulolytic bacteria and hydrogenotrophic methanogens would be the dominant reaction in the thermophilic methanogenic reactor degrading cellulosic materials.     

两级厌氧工艺处理酒精废液的工业化研究

研究了工业规模两级上流式厌氧污泥床反应器（３００ｍ３）在不同温度、不同运行方式下对酒精废液的处理效能．结果表明,将一级高温上流式厌氧污泥床与一级中温上流式厌氧污泥床反应器串联,系统出水水质最佳．当进水ＣＯＤ值为２１０００～２５７００ｍｇ／Ｌ,有机负荷７ｋｇ／（ｍ３·ｄ）左右,系统出水ＣＯＤ值可降低到１５００ｍｇ／Ｌ以下     

Membrane-aerated biofilm reactor for the treatment of acetonitrile wastewater

A membrane-aerated biofilm reactor (MABR) was studied for the treatment of wastewater containing acetonitrile, a typical organonitrile compound. The MABR used hydrophobic hollow fiber membranes as the diffusers for bubbleless aeration as well as the carriers for biofilm growth. The objectives were to prevent the stripping-loss of acetonitrile during aeration and to achieve acetonitrile biodegradation plus nitrogen removal simultaneously in a single biolfilm on the membranes. In the MABR, oxygen and substrates were supplied to the biofilm from opposite sides, in contrast to those from the same side in conventional biofilm bioreactors. Operational factors, including surface loading rate and upflow fluid velocity in the bioreactor, on the effect of acetonitrile biodegradation performance were examined. The profiles of dissolved oxygen concentration and microbial activities and populations in the biofilm were investigated. Experimental results showed that, with the adapted microorganisms, removal of acetonitrile at approximately 98.6 and 83.3%, in terms of total organic carbon and total nitrogen, were achieved at a surface loading rate (in terms of membrane surface) of up to 11.29 g acetonitrile/ m2 x d with an upflow fluid velocity of 12 cm/s and a hydraulic retention time of 30 h. The biofilm on the membranes developed an average thickness of about 1.6 mm in the steady state and consisted of oxic/anoxic/anaerobic zones that provided different functions for acetonitrile degradation, nitrification, and denitrification. The acetonitrile-degrading bacteria in the MABR appeared to secrete more extracellular polymeric substances that enhanced the attachment and development of the biofilm on the membranes. The study demonstrated the potential of using the MABR for the treatment of organonitrile wastewater.     

Effect of temperature on microbial community of a glucose-degrading methanogenic consortium under hyperthermophilic chemostat cultivation

We continuously fed an anaerobic chemostat with synthetic wastewater containing glucose as the sole source of carbon and energy to study the effects of temperature on the microbial community under hyperthermophilic (65-80 degrees C) conditions. Methane was produced normally up to 77.5 degrees C at a dilution rate of 0.025 d(-1). However, the concentration of microorganisms and the rate of gas production decreased with increasing operation temperature. The microbial community in the chemostat at various temperatures was analyzed based on the 16S rRNA gene using molecular biological techniques including clone library analysis and denaturing gradient gel electrophoresis (DGGE). Aceticlastic methanogens related to Methanosarcina thermophila were detected at 65 degrees C and hydrogenotrophilic methanogens related to Methanothermobacter thermautotrophicus were the dominant methanogens between 70 degrees C to 77.5 degrees C. Bacteria related to Clostridium stercorarium and Thermoanaerobacter subterraneus comprised the dominant glucose-fermenting bacteria at temperatures of 65 degrees C and above, respectively. Bacteria related to Thermacetogenium phaeum and to Tepidiphilus margaritifer and Petrobacter succinatimandens were the dominant acetate-oxidizing bacteria at 70 degrees C and at 75-77.5 degrees C, respectively. The results suggested that, at temperatures of 70 degrees C and above, methane production via the aceticlastic pathway was negligible and indirect methanogenesis from acetate was dominant. Since acetate oxidation is a rate limiting step and a higher temperature favors the hydrolysis and acid formation, a two stage fermentation process, acidogenic and methanogenic fermentation stages operated under different temperatures, should be more suitable for the thermophilic anaerobic treatment at temperatures above 65 degrees C.     

Syntrophic degradation of proteinaceous materials by the thermophilic strains Coprothermobacter proteolyticus and Methanothermobacter thermautotrophicus

Protein is a major component of organic solid wastes, and therefore, it is necessary to further elucidate thermophilic protein degradation process. The effects of hydrogenotrophic methanogens on protein degradation were investigated using the proteolytic bacterial strain CT-1 that was isolated from a methanogenic thermophilic (55°C) packed-bed reactor degrading artificial garbage slurry. Strain CT-1 was closely related to Coprothermobacter proteolyticus, which is frequently found in methanogenic reactors degrading organic solid wastes. Strain CT-1 was cultivated in the absence or presence of Methanothermobacter thermautotrophicus by using 3 kinds of proteinaceous substrates. Degradation rates of casein, gelatin, and bovine serum albumin were higher in co-cultures than in monocultures. Strain CT-1 showed faster growth in co-cultures than in monocultures. M. thermautotrophicus comprised 5.5-6.0% of the total cells in co-culture. Increased production of ammonia and acetate was observed in co-cultures than in monocultures, suggesting that addition of M. thermautotrophicus increases the products of protein degradation. Hydrogen produced in the monocultures was converted to methane in co-cultures. These results suggest that thermophilic proteolytic bacteria find it favorable to syntrophically degrade protein in a methanogenic environment, and that it is important to retain hydrogen-scavenging methanogens within the reactor.     

Microbial analyses by fluorescence in situ hybridization of well-settled granular sludge in brewery wastewater treatment plants

The characteristics of granular sludge from full-scale upflow anaerobic sludge blanket reactors used for the treatment of brewery wastewater were investigated. Fluorescence in situ hybridization (FISH) analyses of settled granules from a reactor that had been treating brewery wastewater stably at COD removal rates of over 90% for more than 6 months showed that a methanogen of the genus Methanosaeta was predominant near the granule surface and that Bacteria were not abundant. The center of the granules was composed of dead or resting cells, or both, which were used as a support for active archaeal and bacterial cells near the surface. Periodic analysis of granules from full-scale plants showed that granules containing methanogens deep within them tended to float. Granules with a Bacteria layer on the surface also tended to float. On the basis of these findings, well-settled granules are considered to have methanogens that develop near the granule surface so that the gases generated during methane fermentation are readily released.     

The anaerobic conversion of methanol under thermophilic conditions: pH and bicarbonate dependence

The thermophilic (55 degrees C) anaerobic conversion of methanol was studied in an unbuffered medium (pH 4+/-0.2) and in a phosphate buffered medium (pH 6.4+/-0.1), in both cases without bicarbonate addition. Our cultivated sludge consortium was unable to degrade methanol under acidic conditions. During the 160 d of continuous operation of an up-flow anaerobic sludge blanket (UASB) reactor (R1), at an organic loading rate (ORL) of 6 gCOD/(l.d) and pH around 4, only 5% of the applied methanol load was consumed and no methane (CH4) was detected. However, hydrogenotrophic methanogens were found to be resistant to exposure to such conditions. At the end of the trial, the hydrogenotrophic methanogenic activity of the sludge was 1.23+/-0.16 gCOD/(gVSS.d) at neutral pH. With methanol as the test substrate, the addition of bicarbonate led to acetate accumulation. A second reactor (R2) was operated for 303 d at OLRs ranging from 5.5 to 25.4 gCOD/(l.d) in order to assess the conversion of methanol at neutral pH (phosphate buffered) in a bicarbonate deprived medium. The reactor performance was poor with a methanol-COD removal capacity limited to about 9.5 gCOD/(l.d). The system appeared to be quite susceptible to any type of disturbance, even at low OLR. The fraction of methanol-COD converted to CH4 and acetate was found to be unaffected by the OLR applied. At the end of the trial, the outcome of the competition was about 50% methanogenesis and 50% homoacetogenesis.     

Wastewater treatment and poly-beta-hydroxybutyrate production using lighted upflow anaerobic sludge blanket method

We investigated the performance of a lighted upflow anaerobic sludge blanket (LUASB) reactor for wastewater treatment and poly-beta-hydroxybutyrate (PHB) production. Phototrophic bacteria were induced from UASB (upflow anaerobic sludge blanket) granules under light conditions (100 microE.m(-2).s(-1)). The ammonium and phosphate ion removal efficiencies of the LUASB reactor were higher than those of the UASB reactor. The difference in the results from runs under light and dark conditions suggested that the ammonium and phosphate ion removal efficiencies were improved by increasing the amount of phototrophic bacteria in the LUASB reactor. The average production rate of PHB from the biomass in the effluent from the LUASB reactor was 6.6-14.0 mg.l(-1)-reactor.d(-1) using acetate-based media and the average PHB content based on the dry bacterial biomass was 15.1-25.3%. The PHB concentration increased by reincubation of the effluent from the LUASB reactor with sodium acetate under light conditions. The UASB granular sludge can decompose a variety of organic substances and in addition the LUASB method can remove ammonium and phosphate ions. The LUASB method thus appears to be appropriate for wastewater treatment and production of phototrophic bacteria and PHB from various wastewaters.     

Microbial community in methanogenic packed-bed reactor successfully operating at short hydraulic retention time

The microbial community in a thermophilic anaerobic packed-bed reactor, which had been successfully operated to convert acetic and butyric acids to methane at a short hydraulic retention time (from 24 h to 1.9 h), was investigated. Archaea closely related to known methanogens were detected by 16S rRNA gene analyses of the effluents, together with diverse types of unidentified bacteria.     

High nitrogen removal performance at moderately low temperature utilizing anaerobic ammonium oxidation reactions

High rates of nitrogen removal from wastewater have been reported using anammox bacteria at temperatures around 37 degrees C, but not at moderately low temperatures. In this study, nitrogen removal performance of an anaerobic biological filtrated (ABF) reactor, filled with porous polyester nonwoven fabric carriers as a fixed bed for anammox bacteria, was tested at 37 degrees C and at moderately low temperature (20-22 degrees C). To attain higher nitrogen removal performance, effects of influent nitrogen concentrations and hydraulic retention time (HRT) on nitrogen removal rates were investigated. Nitrogen removal rate increased with influent ammonium and nitrite concentrations, resulting in a removal rate of 3.3 kg-N/m(3)/d on day 32 for an HRT of 180 min at 37 degrees C. However, influent nitrite concentrations greater than 280 mg/l inhibited anammox activity. Therefore, the influent nitrite concentration was adjusted to be below 280 mg/l, and high-loading tests were performed for a shorter HRT. As a result, a nitrogen conversion rate of 11.5 kg-N/m(3)/d was achieved. Moreover, to evaluate long-term anammox activity at moderately low temperatures, ABF reactors were operated for 446 d. Anammox activity could be maintained at 20-22 degrees C, and stable nitrogen removal performance was observed. Furthermore, high nitrogen conversion rate of 8.1 kg-N/m(3)/d was attained. These results clearly show that an appropriate nitrite concentration in the influent and a shorter HRT resulted in high nitrogen conversion rates. The nitrogen removal performance we obtained at moderately low temperatures will open the door for application of anammox processes to many types of industrial wastewater treatment.     

Effects of temperature and hydraulic retention time on anaerobic digestion of food waste

A modified three-stage methane fermentation system was developed to digest food waste efficiently. This system consisted of three stages: semianaerobic hydrolysis, anaerobic acidogenesis and strictly anaerobic methanogenesis. In this study, we examined the effects of temperature and hydraulic retention time (HRT) on the methanogenesis. Operation temperature was adjusted from 30 degrees C to 55 degrees C, and the HRTs ranged from 8 to 12 d. The rate of soluble chemical oxygen demand (sCOD) removal correlated with digestion time according to the first-order kinetic model developed by Grau et al. [Water Res., 9, 637-642 (1975)]. With liquor food waste, thermophilic digesters showed a higher rate of sCOD removal than mesophilic digesters. The rates of biogas and methane production by thermophilic digesters were higher than those by mesophilic digesters regardless of HRT. Although maximum biogas production occurred when an HRT of 10 d was used, the methane yield was the highest in the reactor when an HRT of 12 d was used (223 l CH4/kg sCODdegraded). However, digestion stability decreased when an HRT of 8 d was used. The concentration of NH3-N generated in this experiment did not inhibit anaerobic digestion.     

Methanogenic pathway and community structure in a thermophilic anaerobic digestion process of organic solid waste

The methanogenic pathway and microbial community in a thermophilic anaerobic digestion process of organic solid waste were investigated in a continuous-flow stirred-tank reactor using artificial garbage slurry as a feedstock. The decomposition pathway of acetate, a significant precursor of CH₄ and a key intermediate metabolite in the anaerobic digestion process, was analyzed by using stable isotopes. A tracer experiment using ¹³C-labeled acetate revealed that approximately 80% of the acetate was decomposed via a non-aceticlastic oxidative pathway, whereas the remainder was converted to methane via an aceticlastic pathway. Archaeal 16S rRNA analyses demonstrated that the hydrogenotrophic methanogens Methanoculleus spp. accounted for > 90% of detected methanogens, and the aceticlastic methanogens Methanosarcina spp. were the minor constituents. The clone library targeting bacterial 16S rRNA indicated the predominance of the novel Thermotogales bacterium (relative abundance: ~ 53%), which is related to anaerobic acetate oxidizer Thermotoga lettingae TMO, although the sequence similarity was low. Uncultured bacteria that phylogenetically belong to municipal solid waste cluster I were also predominant in the microflora (~ 30%). These results imply that the microbial community in the thermophilic degrading process of organic solid waste consists exclusively of unidentified bacteria, which efficiently remove acetate through a non-aceticlastic oxidative pathway.     

Methane fermentation of bean curd refuse

The methane fermentation of bean curd refuse was studied in a 1 l reactor with a draft tube which was operated in a fed-batch mode with a once-a-day feeding cycle using two kinds of methanogens. The effects of substrate loading rate on the methane yield and on the behaviors of the product species were examined. A unified approach was employed for the material balance between the substrate (reactant) and product species based on the elemental carbon content in the species being considered. The classified product species are methane and carbon dioxide in the gas phase, soluble TOCs excluding VFAs, ICs and VFAs in the liquid phase and cells, and the unconverted substrate in the solid phase. It is found that the methane yield increases with the increasing substrate loading rate, reaches a maximum and then decreases with a further increasing loading rate. The maximum methane yield is found to be as high as 53.7% which is very close to the theoretical yield of 55%. It is also found that there is a critical substrate loading rate beyond which the operation becomes impossible due to excessive accumulation of unconverted solids.     

Nitrogen removal via nitrite from municipal wastewater at low temperatures using real-time control to optimize nitrifying communities

Although many studies regarding nitrogen removal via nitrite have been carried out, very limited research has been undertaken on nitrogen removal via nitrite at low temperatures. In this study, to improve the nitrogen removal efficiency from municipal wastewater, a pilot-plant of sequencing batch reactor with a working volume of 54 m3 was used to investigate nitrogen removal via nitrite from municipal wastewater at normal and low water temperature. The obtained results showed that high nitrogen removal efficiency with effluent total nitrogen below 3 mg/L could be achieved. Using real-time control with temperature ranging from 11.9 to 26.5 degrees C under normal dissolved oxygen condition (> or =2.5 mg/L), nitrogen removal via nitrite was successfully and stably achieved for a long period (180 days) with average nitrite accumulation rate above 95%. Fluorescence in situ hybridization was carried out to investigate the quantitative changes of nitrifying microbial community in the activated sludge. Fluorescence in situ hybridization results approved that the nitrifying microbial communities were optimized; ammonia oxidizing bacteria became the dominant nitrifying bacteria and nitrite oxidizing bacteria had been washed out of the activated sludge.     

Nitrogen removal characteristics and biofilm analysis of a membrane-aerated biofilm reactor applicable to high-strength nitrogenous wastewater treatment

A membrane-aerated biofilm reactor (MABR) capable of simultaneous nitrification and denitrification in a single reactor vessel was developed to investigate the characteristics of nitrogen removal from high-strength nitrogenous wastewater, and biofilm analysis using microelectrodes and the fluorescence in situ hybridization (FISH) technique was performed. Mean removal percentages of total organic carbon (TOC) and nitrogen were 96% and 83% at removal rates of 5.76 g-C m(-2) d(-1) and 4.48 g-N m(-2) d(-1), respectively. For stable removal efficiency, constant washing of the biofilm was needed. Dissolved oxygen microelectrode measurement revealed that the biofilm thickness was about 1600 microm, and that oxygen penetrated about 300 to 700 microm, from the outer surface of the membrane. Furthermore, FISH analysis revealed that ammonia-oxidizing bacteria (AOB) were located near the outer surface of the membrane, whereas other bacteria were located from the inner to the outer part of the biofilm. Combining these results demonstrated that simultaneous nitrification and denitrification occurred in the biofilm of the MABR system. In addition, stoichiometric analysis revealed that after 130 d(-1), the free ammonia (FA) concentration ranged within the concentration causing inhibition of the growth of nitrite oxidizing bacteria (NOB) and that AOB consumed 86% of the oxygen supplied through the intra-membrane. These results indicate that nitrogen removal not via nitrate but via nitrite was mainly achieved in the MABR system.     

生物促生剂、解毒剂在造纸废水处理中的应用

基于废水处理系统受到上游改变纸品生产工艺的影响,造成废水处理系统中活性污泥活性不强或死亡等问题,在造纸废水处理生化段使用生物促生剂Bio-energizer(BE)、解毒剂Micatrol(MT)。结果表明:受损的造纸废水处理系统在使用BE、MT 30d时间内,剂量仅为1mg/kg,能够使系统恢复。在不增加任何基建投入的情况下,使用BE/MT后,系统COD去除率能够达到89.3%,出水COD能够稳定在100mg/l以下。使用BE/MT后,系统中微生物种类丰富,以钟虫等固着型纤毛虫类居多,出现了轮虫等后生动物,总体呈现多样化的趋势。     

Formate dehydrogenase in rice plant: growth stimulation effect of formate in rice plant

It was previously suggested that a population of phototrophic bacteria, Rhodopseudomonas palustris strain RN1 and Blastochloris sulfoviridis strain GN1 could be induced from granules in a lighted upflow anaerobic sludge blanket (LUASB) reactor. The present study showed that both strains RN1 and GN1 could use acetate, propionate, butyrate, and lactate as electron donors under anaerobic light conditions. The composition of organic acids in the effluent from the LUASB reactor was studied to investigate competitive consumption between acetogenic bacteria, methanogens, and phototrophic bacteria in the reactor. When acetate, propionate and lactate were supplied to the reactor, a small amount of acetate and propionate was observed in the effluent under light conditions. The concentrations of acetate and propionate increased under dark conditions compared with those under light conditions using organic acid and peptone media as the influent. When starch was supplied to the reactor, the concentrations of formate, acetate, propionate, butyrate, and lactate in the effluent were less than 0.5 mg C.l(-1) during operation under light and dark conditions. The concentrations of ammonium and phosphate in the effluent under dark conditions were higher than those under light conditions. These results suggested that phototrophic bacteria in the LUASB reactor consumed acetate and propionate as well as ammonium and phosphate in competition with methanogens and acetogenic bacteria.