Anaerobic digestion of sludge by different pretreatments: Changes of amino acids and microbial community

•Tryptophan protein, and aromatic protein I/II were the key identified proteins. •Cysteine was more correlated with methane production than other amino acids. •The presence of cysteine can promote methane production and degradation of VFAs. •The presence of cysteine can lower ORP and increase biomass activity. •Predominant Tissierella and Proteiniphilum were noted in pretreated sludge samples. Many studies have investigated the effects of different pretreatments on the performance of anaerobic digestion of sludge. However, the detailed changes of dissolved organic nitrogen, particularly the release behavior of proteins and the byproducts of protein hydrolysis-amino acids, are rarely known during anaerobic digestion of sludge by different pretreatments. Here we quantified the changes of three types of proteins and 17 types of amino acids in sludge samples solubilized by ultrasonic, thermal, and acid/alkaline pretreatments and their transformation during anaerobic digestion of sludge. Tryptophan protein, aromatic protein I, aromatic protein II, and cysteine were identified as the key dissolved organic nitrogen responsible for methane production during anaerobic digestion of sludge, regardless of the different pretreatment methods. Different from the depletion of other amino acids, cysteine was resistant to degradation after an incubation period of 30 days in all sludge samples. Meanwhile, the “cysteine and methionine metabolism (K00270)” was absent in all sludge samples by identifying 6755 Kyoto Encyclopedia of Genes and Genomes assignments of genes hits. Cysteine contributed to the generation of methane and the degradation of acetic, propionic, and n-butyric acids through decreasing oxidation-reduction potential and enhancing biomass activity. This study provided an alternative strategy to enhance anaerobic digestion of sludge through in situ production of cysteine.     

添加蚓粪对玉米秸秆厌氧发酵产气特性的影响

以玉米秸秆为原料,总固体(TS)含量为50g.kg-1,采用中温35℃批式发酵,考察了添加蚓粪对玉米秸秆厌氧发酵产气的影响。结果表明:玉米秸秆单独发酵时,单位TS产气量为333.3mL.g-1,甲烷产量为202.0mL.g-1,并在反应初期出现酸化现象;蚓粪与玉米秸秆TS质量比为1:1进行发酵时,单位TS产气量为400.0mL.g-1,甲烷产量为269.6mL.g-1,相对于玉米秸秆单独厌氧发酵分别提高了20.0%和35.5%,且反应过程中未出现酸化现象。说明蚓粪适合作为玉米秸秆厌氧发酵产气的添加剂。     

Enhanced methane production during long-term UASB operation at high organic loads as enabled by the immobilized Fungi

• Fungi enable the constant UASB operation even at OLR of 25.0 kg/(m³×d). • The COD removal of 85.9% and methane production of 5.6 m³/(m³×d) are achieved. • Fungi inhibit VFAs accumulation and favor EPS generation and sludge granulation. • Fungi enrich methanogenic archaea and promote methanogenic pathways. Anaerobic digestion is widely applied in organic wastewater treatment coupled with bioenergy production, and how to stabilize its work at the high organic loading rate (OLR) remains a challenge. Herein, we proposed a new strategy to address this issue via involving the synergetic role of the Aspergillus sydowii 8L-9-F02 immobilized beads (AEBs). A long-term (210-day) continuous-mode operation indicated that the upflow anaerobic sludge bed (UASB) reactor (R1, with AEBs added) could achieve the OLR as high as 25.0 kg/(m³×d), whereas the control reactor (R0, with AEBs free) could only tolerate the maximum OLR of 13.3 kg/(m³×d). Remarkably, much higher COD removal (85.9% vs 23.9%) and methane production (5.4 m³/(m³×d) vs 2.2 m³/(m³×d)) were achieved in R1 than R0 at the OLR of 25.0 kg/(m³×d). Such favorable effect results from the facts that fungi inhibit VFAs accumulation, favor the pH stabilization, promote the generation of more extracellular polymeric substance, and enhance the sludge granulation and settleability. Moreover, fungi may enhance the secretion of acetyl-coenzyme A, a key compound in converting organic matters to CO₂. In addition, fungi are favorable to enrich methanogenic archaea even at high OLR, improving the activity of acetate kinase and coenzyme F₄₂₀ for more efficient methanogenic pathway. This work may shed new light on how to achieve higher OLR and methane production in anaerobic digestion of wastewater.     

微波碱解处理剩余污泥的厌氧消化性能

为了研究微波碱解预处理（microwave/NaOH pretreatment,MNP）剩余污泥在高温和中温条件下厌氧消化的性能,采用半连续完全混合式反应器分别研究了MNP处理后的剩余污泥在高温厌氧消化工艺（thermophilic anaerobic di-gestion process,TADP）和中温厌氧消化工艺...     

添加剂对污泥厌氧消化性能的影响

在间歇培养条件下,研究了还原型辅酶Ⅱ（NADPH）、乙酰辅酶A（Acetyl Co A）和对氨基苯甲酸（PABA）3种添加剂对污泥厌氧消化性能的影响.结果表明,3种微生物活性促进剂均能促进污泥厌氧消化产气.其中,NADPH的促进效果最为显著,消化第35 d,产甲烷量比对照组高15.90%.在污泥含固率为3%、未调初始pH（pH=6.7）和温度35℃的厌氧消化条件下,NADPH的最佳添加量为50 mg.L-1,消化第36 d,污泥累积产甲烷量127.13 mL.g-1VSS.在含固率3%、初始pH=8.5、温度55℃和NADPH添加量为50 mg.L-1的工艺条件下,污泥厌氧产气效果最佳,消化第30 d时累积产甲烷量达158.02 mL.g-1VSS.     

Increasing the performance of anaerobic digestion: Pilot scale experimental study for thermal hydrolysis of mixed sludge

The performance of a pilot plant operation combining thermal hydrolysis (170°C, 30 min) and anaerobic digestion (AD) was studied, determining the main properties for samples of fresh mixed sludge, hydrolyzed sludge, and digested sludge, in order to quantify the thermal pretreatment performance (disintegration, solubilisation, and dewaterability) and its impact on the anaerobic digestion performance (biodegradability, volatile solids reduction, and digester rheology) and end product characteristics (dewaterability, sanitation, organic and nitrogen content). The disintegration achieved during the thermal treatment enhances the sludge centrifugation, allowing a 70% higher total solids concentration in the feed to anaerobic digestion. The digestion of this sludge generates 40% more biogas in half the time, due to the higher solids removal compared to a conventional digester. The waste generated can be dewatered by centrifugation to 7% dry solids without polymer addition, and is pathogen free.     

厌氧条件下剩余污泥中磷及相关指标的释放和变化规律

污水处理过程中产生的剩余污泥富含大量的氮磷元素,从剩余污泥中回收磷是解决磷资源日益缺乏的一种有效途径。探寻出剩余污泥中磷的释放规律是实现剩余污泥中磷回收的首要前提。因此,以实际污水处理厂污泥为研究对象,建立污泥停留时间为5d的中试模型系统。通过系统分析5d停留时间的厌氧条件下污泥中污泥浓度、上清液总磷和氨氮浓度的变化情况,为后续的污泥磷回收提供支撑条件。研究结果表明,在中试系统污泥停留时间5d的厌氧条件下,剩余污泥微生物衰亡自溶或被分解,胞内物质释放,从而使固态物质转化为液态,污泥中磷及相关的氮等物质得到了较大的释放,污泥上清液总磷和氨氮质量浓度可分别达到100和40 mg·L^-1以上。所释放出的氮磷浓度足以满足鸟粪石回收氮磷方法所需的最低经济性要求,为污泥进行厌氧消化后采用鸟粪石的方法回收释放的氮磷提供了重要的基础依据。研究中还发现5d停留时间下, SS和VSS都有不同程度的降低,二者分别减少8.34%和10.14%以上,其中VSS的减少量占SS减少量的65%左右。同时,进入厌氧反应系统的初始污泥浓度对于氮磷的释放有着较大的影响,反应系统的SS在6300~7200 mg·L^-1的条件下,磷和氮的单位质量污泥释放量达到最佳,分别达到单位干污泥0.015和0.006 mg·mg^-1。研究结果为剩余污泥中回收氮磷提供了重要的依据。     

Use of gamma-irradiation pretreatment for enhancement of anaerobic digestibility of sewage sludge

The effects of γ-irradiation pretreatment on anaerobic digestibility of sewage sludge was investigated in this paper. Parameters like solid components, soluble components, and biogas production of anaerobic digestion experiment for sewage sludge were measured. The values of these parameters were compared before and after γ-irradiation pretreatment. Total solid (TS), volatile solid (VS), suspended solid (SS), volatile suspended solid (VSS), and average floc size of samples decreased after γ-irradiation treatment. Besides, floc size distribution of sewage sludge shifted from 80–100 μm to 0–40 μm after γ-irradiation treatment at the doses from 0 to 30 kGy, which indicated the disintegration of sewage sludge. Moreover, microbe cells of sewage sludge were ruptured by γ-irradiation treatment, which resulted in the release of cytoplasm and increase of soluble chemical oxygen demand (SCOD). Both sludge disintegration and microbe cells rupture enhanced the subsequent anaerobic digestion process, which was demonstrated by the increase of accumulated biogas production. Compared with digesters fed with none irradiated sludge, the accumulated biogas production increased 44, 98, and 178 mL for digesters fed sludge irradiated at 2.48, 6.51, and 11.24 kGy, respectively. The results indicated that γ-irradiation pretreatment could effectively enhance anaerobic digestibility of sewage sludge, and correspondingly, could accelerate hydrolysis process, shorten sludge retention time of sludge anaerobic digestion process.     

猪场废水厌氧消化过程中的除磷效果

采用序批式半连续厌氧消化试验方法,研究猪场废水厌氧消化过程中磷的去除情况。结果表明,水力停留时间（HRT）为1、3、6和9 d的厌氧反应器平均除磷率分别为65.0%、81.1%、82.7%和83.0%,而COD平均去除率分别为54.5%、82.3%、87.0%和85.9%。厌氧反应器除磷能力随沼气产量的增加而增加,说明厌氧反应器中磷的去除与产甲烷过程密切相关。对厌氧消化前后的污泥进行浸提后发现,厌氧消化过程中,化学反应生成磷酸盐沉淀的除磷作用十分显著,污泥中正磷酸盐,与铁结合的磷化合物（Fe-RP）,还原可溶性磷,与钙、镁离子结合的磷化合物（Ca-RP、Mg-RP）以及无机或有机聚合磷增加量分别为0.027 8～0.101 5、0.013 5～0.081 0、0.2165～0.430 5、23.4～54.8和7.2～21.5 mg.g-1;且总体而言,HRT越长,污泥中与不同金属结合的磷增加量就越大。从HRT、磷和有机物的去除效果以及沼气产气速率3个方面综合考虑,猪场废水厌氧消化反应器的HRT控制在3 d为宜。     

厌氧好氧一体化生物反应器处理发酵废水的研究

采用新型厌氧好氧一体化生物反应器对发酵废水进行了中试处理研究.试验结果表明,系统总有机负荷最高可达到8.88kg(COD)m-3d-1,系统去除率稳定在88.10%~96.88%,说明反应器处理效率高,抗冲击能力强.反应器结构合理,利于保持丰富、高活性的微生物,反应器厌氧区颗粒污泥TS高达83.9gL-1,VS/TS为56.9%~57.4%,比产甲烷活性为280~350mL(CH4)gvss-1d-1;好氧区固定化微生物TS高达64.03gL-1载体,VS/TS为94.02%~94.30%.反应器各功能区对废水的降解过程分析,说明反应器厌氧区和好氧区一体化结构合理,可将废水逐级降解,从而保证整个系统的处理效果.图8表4参11     

Roles of acid-producing bacteria in anaerobic digestion of waste activated sludge

Three acid-producing strains, AFB-1, AFB-2 and AFB-3, were isolated during this study, and their roles in anaerobic digestion of waste activated sludge (WAS) were evaluated. Data of 16S rRNA method showed that AFB-1 and AFB-2 were Bacillus coagulans, and AFB-3 was Escherichia coli. The removal in terms of volatile solids (VS) and total chemical oxygen demand (TCOD) was maximized at 42.7% and 44.7% by inoculating Bacillus coagulans AFB-1. Besides, the optimal inoculum concentration of Bacillus coagulans AFB-1 was 30% (v/v). Solubilization degree experiments indicated that solubilization ratios (SR) of WAS reached 20.8%±2.2%, 17.7%±1.48%, and 11.1%±1.53%. Volatile fatty acids (VFAs) concentrations and compositions were also explored with a gas chromatograph. The results showed that VFAs improved by 98.5%, 53.0% and 11.6% than those of the control, respectively. Biochemical methane potential (BMP) experiments revealed that biogas production increased by 90.7% and 75.3% when inoculating with Bacillus coagulans AFB-1 and AFB-2. These results confirmed that the isolated acid-producing bacteria, especially Bacillus coagulans, was a good candidate for anaerobic digestion of WAS.

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Developing “precise-acting” strategies for improving anaerobic methanogenesis of organic waste: Insights from the electron transfer system of syntrophic partners

Methanogenesis is the last step in anaerobic digestion, which is usually a rate-limiting step in the biological treatment of organic waste. The low methanogenesis efficiency (low methane production rate, low methane yield, low methane content) substantially limits the development of anaerobic digestion technology. Traditional pretreatment methods and bio-stimulation strategies have impacts on the entire anaerobic system and cannot directly enhance methanogenesis in a targeted manner, which was defined as “broad-acting” strategies in this perspective. Further, we discussed our opinion of methanogenesis process with insights from the electron transfer system of syntrophic partners and provided potential targeted enhancing strategy for high-efficiency electron transfer system. These “precise-acting” strategies are expected to achieve an efficient methanogenesis process and enhance the bio-energy recovery of organic waste.     

Experimental study of nitrite accumulation in predenitrification biological nitrogen removal process

The effect of dissolved oxygen (DO) concentration on nitrite accumulation was investigated in a pilot-scale pre-denitrification process at room temperature for 100 days. In the first 10 days, due to the instability of the system, the DO concentration fluctuated between 1.0 and 2.0 mg/L. In the next 14 days, the DO concentration was kept at 0.5 mg/L and nitrite accumulation occurred, with the average nitrite accumulation rate at 91%. From the 25th day, the DO concentration was increased to 2.0 mg/L to destroy the nitrite accumulation, but nitrite accumulation rate was still as high as 90%. From the 38th day the nitrite accumulation rate decreased to 15%–30% linearly. From the 50th day, DO concentration was decreased to 0.5 mg/L to resume nitrite accumulation. Until the 83rd day the nitrite accumulation rate began to increase to 80%. Dissolved oxygen was the main cause of nitrite accumulation, taking into account other factors such as pH, free ammonia concentration, temperature, and sludge retention time. Because of the different affinity for oxygen between nitrite oxidizing bacteria and ammonia oxidizing bacteria when DO concentration was kept at 0.5 mg/L, nitrite accumulation occurred.     

Treatment of spent wash in anaerobic mesophilic suspended growth reactor (AMSGR)

Approximately 400 KL of spent wash or vinasse per annum is generated at an average COD concentration of 100,000 mg/l, by over 250 distilleries in India. There is an urgent need to develop, assess and use ecofriendly methods for the disposal of this high strength wastewater. Therefore, an attempt was made to investigate a few aspects of anaerobic digestion of spent wash collected from a distillery. The study was carried out in a 4 L laboratory scale anaerobic mesophilic suspended growth reactor. After the successful startup, the organic loading was increased stepwise to assess the performance of the reactor. During the study period, biogas generated was recorded and the maximum gas generated was found to be 16.9 L at an Organic Loading Rate (OLR) of 38 g COD/L. A 500% increase in the Volatile Fatty Acid (VFA) concentration (2150 mg/L) was observed, when the OLR was increased from 38 to 39 g COD/L. During the souring phase the removal of COD, Total Solids (TS) and Volatile Solids (VS) were in the order of 52%, 40% and 46% respectively. The methane content in the biogas varied from 65% to 75%.     

四环素类抗生素和铜复合污染对猪粪厌氧消化的影响

兽用抗生素和矿物元素添加剂可起到预防动物疾病、促进动物生长、提高饲料转化率等作用,因此被广泛应用于畜禽养殖业。本研究以猪粪中温厌氧消化为研究目标,采用全自动甲烷潜势测试系统,考察了一定浓度的四环素(TC:30 mg·kg~(-1)dry weight,DW)、土霉素(OTC:50 mg·kg~(-1)DW)和金霉素(CTC:15 mg·kg~(-1)DW)对厌氧累积产甲烷量和日产甲烷速率的影响。结果表明,TC、OTC和CTC对猪粪中温厌氧消化累积产甲烷量和日产甲烷速率均有促进作用(累积产甲烷总量提高比例分别为7.9%、0.4%和5.4%)。另外,采用超高效液相色谱-四极杆串联质谱对猪粪厌氧消化前后样品中四环素类抗生素及其代谢产物进行了分析。结果表明,液相中的四环素类抗生素在猪粪厌氧消化过程中得到了明显的去除,去除率达到90%~100%;而固相中只有金霉素和差向异构金霉素有明显的去除效果,去除率分别为41.69%和41.58%。采用Tessier连续提取法对猪粪厌氧消化前后样品中5种形态的铜包括可交换态、碳酸盐结合态、铁锰氧化结合态、有机物结合态、残渣态进行了分析,结果表明,猪粪厌氧消化后,可交换态、碳酸盐结合态和铁锰结合态的铜浓度比厌氧消化前分别降低了1%~9%、0.1%~3%、12%~19%,而有机态和残渣态的铜浓度却在厌氧消化后分别增加了15%~35%、1%~2%。厌氧消化后,70%~80%的铜都是以有机铜的形态存在。铜逐渐从不稳定态转化为相对稳定的有机态和残渣态铜,因此,厌氧消化过程使铜从可生物利用态转变为不可生物利用态,趋于稳定化。     

Comparison of different valent iron on anaerobic sludge digestion: Focusing on oxidation reduction potential,dissolved organic nitrogen and microbial community

• ORP value from −278.71 to −379.80 mV showed indiscernible effects on methane yield. • Fe(II) and Fe(III) promoted more degradation of proteins and amino acids than Fe⁰. • The highest enrichment of Geobacter was noted in samples added with Fe⁰. • Cysteine was accumulated during iron enhanced anaerobic sludge digestion. • Both iron content and valence were important for methane production. This study compared effects of three different valent iron (Fe⁰, Fe(II) and Fe(III)) on enhanced anaerobic sludge digestion, focusing on the changes of oxidation reduction potential (ORP), dissolved organic nitrogen (DON), and microbial community. Under the same iron dose in range of 0−160 mg/L after an incubation period of 30 days (d), the maximum methane production rate of sludge samples dosed with respective Fe⁰, Fe(II) and Fe(III) at the same concentration showed indiscernible differences at each iron dose, regardless of the different iron valence. Moreover, their behavior in changes of ORP, DON and microbial community was different: (1) the addition of Fe⁰ made the ORP of sludge more negative, and the addition of Fe(II) and Fe(III) made the ORP of sludge less negative. However, whether being more or less negative, the changes of ORP may show unobservable effects on methane yield when it ranged from −278.71 to −379.80 mV; (2) the degradation of dissolved organic nitrogen, particularly proteins, was less efficient in sludge samples dosed with Fe⁰ compared with those dosed with Fe(II) and Fe(III) after an incubation period of 30 d. At the same dose of 160 mg/L iron, more cysteine was noted in sludge samples dosed with Fe(II) (30.74 mg/L) and Fe(III) (27.92 mg/L) compared with that dosed with Fe⁰ (21.75 mg/L); (3) Fe⁰ particularly promoted the enrichment of Geobacter, and it was 6 times higher than those in sludge samples dosed with Fe(II) and Fe(III) at the same dose of 160 mg/L iron.     

Anaerobic treatment of wastewater containing methanol in upflow anaerobic sludge bed (UASB) reactor

The direct conversion of methanol into methane is the main process in anaerobic treatment of methanol containing wastewater. However, acetic acid can also be produced from methanol theoretically, which may probably result in an abrupt pH drop and deteriorate the anaerobic process. Therefore, it is interesting to know what would really happen in an anaerobic reactor treating methanol wastewater. In this study, an up-flow anaerobic sludge bed (UASB) reactor treating methanol wastewater was operated. The chemical oxygen demand (COD), acetic acid and pH of the effluent were monitored at different loadings and influent alkalinity. The results showed that the anaerobic reactor could be operated steadily at as low as 119 mg/L of influent alkalinity and high organic loading rate with no obvious pH drops. Volatile fatty acids accumulation was not observed even at strong shock loadings. The microorganisms in the sludge at the end of the test became homogeneous in morphology, which were mainly spherical or spheroidal in shape.     

Degradation of polyacrylamide (PAM) and methane production by mesophilic and thermophilic anaerobic digestion: Effect of temperature and concentration

• PAM degradation in thermophilic AD in comparison with mesophilic AD. • PAM degradation and its impact on thermophilic and mesophilic AD. • Enhanced methane yield in presence of PAM during thermophilic and mesophilic AD. • PAM degradation and microbial community analysis in thermophilic and mesophilic AD. Polyacrylamide (PAM) is generally employed in wastewater treatment processes such as sludge dewatering and therefore exists in the sludge. Furthermore, it degrades slowly and can deteriorate methane yield during anaerobic digestion (AD). The impact or fate of PAM in AD under thermophilic conditions is still unclear. This study mainly focuses on PAM degradation and enhanced methane production from PAM-added sludge during 15 days of thermophilic (55°C) AD compared to mesophilic (35°C) AD. Sludge and PAM dose from 10 to 50 g/kg TSS were used. The results showed that PAM degraded by 76% to 78% with acrylamide (AM) content of 0.2 to 3.3 mg/L in thermophilic AD. However, it degraded only 27% to 30% with AM content of 0.5 to 7.2 mg/L in mesophilic AD. The methane yield was almost 230 to 238.4 mL/g VSS on the 8th day in thermophilic AD but was 115.2 to 128.6 mL/g VSS in mesophilic AD. Mechanism investigation revealed that thermophilic AD with continuous stirring not only enhanced PAM degradation but also boosted the organics release from the sludge with added PAM and gave higher methane yield than mesophilic AD.     

Mechanism on minimization of excess sludge in oxic-settling-anaerobic (OSA) process

The oxic-settling-anaerobic (OSA) process is a promising wastewater treatment technique for efficiently reducing sludge production and improving the stability of process operation. In this paper, the possible factors of sludge reduction such as sludge decay, uncoupled metabolism, and anaerobic oxidation with low sludge production were discussed in the OSA process. It has been confirmed that sludge decay is the decisive cause in the OSA process, accounting for 66.7% of sludge production reduction. Sludge decay includes hydrolysis and acidogenesis of dead microorganisms and particle organic carbon adsorbed in sludge floc and endogenous metabolism. By batch experiments, it has been proven that there is energetic uncoupling in the OSA system since microorganisms were exposed to alternative anaerobic and aerobic environment. It accounts for about 7.5% of sludge production reduction. Soluble chemical oxygen demand (SCOD) released from the anaerobic sludge tank in the OSA process was used as the substrate for cryptic growth. The substrate was used for anoxic denitrifying, anaerobic phosphorus release, sulfate reduction, and methane production. These anaerobic reactions in the sludge anaerobic tank have lower sludge production than in the aerobic oxidation when equivalent SCOD is consumed, which may lead to approximately 23% of sludge reduction in the OSA process. It has been concluded that multiple causes resulted in the minimization of excess sludge in the OSA system. The microbial community structure and diversity of sludge samples from the CAS (conventional activated sludge) and OSA systems were investigated by 16 SrDNA PCR-DG-DGGE (polymerase chain reaction-double gradient-denaturing gradient gel electrophoresis). DGGE profile and cluster analysis showed more abundant species in the OSA system contrasting to microbial communities in the CAS system.     

两相厌氧发酵互花米草的特性

利用两相(固态水解酸化相+湿式发酵产甲烷相)发酵互花米草,通过尿素的添加来调整厌氧发酵互花米草的碳氮比,考察两相厌氧发酵互花米草的特性;并利用FTIR考察水解发酵前后互花米草结构的变化.研究结果表明,由于固态水解渗滤液对尿素的淋滤,产生高浓度的氨氮溶液,抑制了湿式发酵产甲烷反应器中的甲烷菌;通过53 d的两相发酵互花米草,单位挥发性固体(VS)累积产气量仅为98.6 mL.g-1(0℃),转化率是理论产气量的17.5%;游离性氨(FA)浓度超过55 mg.L-1,就会严重抑制高温甲烷菌;红外光谱分析表明,石灰堆沤预处理对互花米草结构改变较小;只有大幅度的强化预处理,才能显著提高互花米草发酵的产气量.