沼气发酵菌的富集培养和应用

1972年以来,我省农村大办沼气的群众运动蓬勃发展,许多生产队、生产大队和人民公社已基本实现沼气化。在大办沼气的群众运动中,我们开展了沼气发酵菌的研究工作。活性污泥的选择为了提高沼气产量并加速产气以及增加沼气中的甲烷成分,我们采集了沼气较多的活性污泥,经富集培养,选取产气量和沼气中甲烷含量较高的污泥作为“菌     

牲畜粪便与麦秆混合厌氧发酵的产气量、发酵时间及最优温度

探索牲畜粪便与作物秸秆混合发酵的产气量和发酵时间与发酵温度之间的关系,是解决农村户用沼气原料选择、确定最优发酵温度和提高农作物秸秆资源化利用效率的关键.采用可控型恒温发酵装置,以猪粪、牛粪和麦秆作为发酵原料,以常温厌氧发酵池的底物为接种物,在总固体（total solid,TS）质量分数为8%的条件下进行批量试验,研究了混合发酵的产气量、发酵时间及最优温度.结果表明：粪便与麦秆混合发酵明显提高了原料的产气效率,其中猪粪与麦秆混合发酵的累积产气量比猪粪作为单一发酵原料高24倍,而牛粪与麦杆混合发酵的累积产气量与单一牛粪无显著差异.猪粪、牛粪与麦秆混合发酵的最优温度均在30 ℃以上,发酵时间在60 d左右.厌氧发酵的发酵时间不总是随着温度的升高而缩短,单一以温度来断定厌氧发酵时间的长短是不可行的.     

高效产氢菌株Enterococcus sp. LG1的分离及产氢特性

采用Hungate厌氧培养技术分别从厌氧污泥、好氧污泥及河底泥中分离出12株厌氧产氢细菌,并对其中的Enterococcus sp.LG1(注册号:EU258743)进行了研究.结果表明,该株细菌为专性厌氧菌,经革兰氏染色结果为阴性.通过16S rDNA碱基测序和比对证实,该菌株是目前尚未报道过的1个新菌种,初步确定其细菌学上的分类地位.同时,以灭菌预处理的污泥为底物培养基,对该菌的产氢能力及污泥发酵过程中底物性质变化(SCOD、可溶性蛋白质、总糖和pH值等)进行了探讨.实验结果显示,产氢茵Enterococcus sp.LG1的发酵过程中只有H2和CO2产生,无CH4产生.产气量最高为36.48 mL/g TCOD,氢气含量高达73.5%,为已报道文献中以污泥为底物发酵制氢中之最高.根据污泥发酵产物分析得知,该菌的发酵类行为典型的丁酸型发酵.     

巴氏杀菌对餐厨垃圾高温厌氧发酵的影响

通过对杀菌、未杀菌餐厨垃圾高温发酵对比试验的研究分析,阐述了餐厨垃圾杀菌处理对发酵的影响。杀菌方式为巴氏消毒法,温度70℃,持续时间10 min,接种物为德青源实验室自行培养的高温发酵菌,底物为餐厨垃圾,有机负荷5 gVS/L,污泥负荷(F/M)为0.5,试验持续32 d,结果显示:杀菌组累计产气量895 mL/gVS,未杀菌组累计产气量795 mL/gVS;杀菌样品产气速率高于未杀菌样品,其VS去除率略低于未杀菌样品。     

预处理对泔脚发酵产氢的强化研究

采用热(80℃,15 min)预处理的城市生活垃圾厌氧消化污泥为接种物,考察了不同预处理方法对泔脚中温(36℃)批式发酵产氢的影响。Gompertz模型拟合结果表明:微波850 W,4 min与pH9.0下预处理泔脚的发酵产氢延迟时间λ、最大比产氢率、产氢率、生物气中氢气的最高体积分数分别为:3.47 h,9.43 mL/(g.h),186.23 mL/g及46.0%时,具有更大的产氢优越性。泔脚的发酵产氢过程也是一个酸化过程,发酵产氢结束后,4个预处理方案的发酵产氢余物的pH在4.40~5.00之间,pH均有较大幅度的下降。     

污泥与餐厨垃圾发酵产氢种泥预处理方法比较

以剩余污泥和餐厨垃圾作为混合基质进行厌氧发酵产氢批式试验,比较六种常用的产氢种泥预处理方法[热处理、化学抑制剂2-溴乙基磺酸钠(BESA)处理、酸处理、碱处理、连续曝气、重复曝气]对产氢的影响。结果表明,未经预处理的种泥氢气产率最低,且有明显的吸氢和产甲烷现象。BESA处理、酸处理、连续曝气和重复曝气种泥产氢效果较好,其中重复曝气预处理种泥氢气产率最高,为86.9 ml-H2/g-VSadded,对产甲烷菌有明显抑制。热处理和碱处理种泥产氢效果较差,反应后期出现吸氢反应并有明显的甲烷累积现象。发酵产氢过程中p H值从中性下降到5.0左右,对产甲烷菌活性也具有一定的抑制作用。     

低温沼气发酵优良菌系筛选及优势菌群分析

【目的】为获得低温沼气发酵高效菌系, 从张家口、承德和邯郸地区采集低温产气良好的沼气池中沼泥样品12份。【方法】以沼泥为接种源进行16 °C?5 °C阶段降温模拟沼气发酵试验, 对处理组HL2、ZG2、CW1及其相应的接种源HLA、ZGB、CWB进行DGGE分析。【结果】ZG2处理组模拟沼气发酵综合性能最优, 与其他处理组呈显著性差异; DGGE图谱显示, 被检测样品中古菌种属多样性丰富, 但图谱中代表优势种属条带的位置存在较大差异。通过16S rDNA克隆及测序分析, 样品中主要优势菌属为甲烷八叠球菌属、甲烷鬃毛菌属和甲烷粒菌属。【结论】DGGE图谱中代表甲烷八叠球菌属的条带是样品ZG2和ZGB中唯一重复出现的条带, 且未作为优势条带出现在其他样品中, 推测甲烷八叠球菌属与低温产沼气有密切相关性。     

产气抑制剂对污泥和食品废弃物混合发酵产酸过程的应用研究

为了提高有机酸的浓度,探讨污泥产酸工业化的可行性,考察了三种产气抑制剂对污泥和食 品废弃物混合发酵的VFA浓度及其组成、底物转化率、产气量及产甲烷势的影响.结果表明 ,采用BES、氯仿和碱(pH10)三种手段可以有效抑制发酵后期产气作用,提高底物产酸转化率,并且对有机酸组成也有一定影响.     

猪羊粪及其配比发酵沼气试验初报

提高农村户用沼气的产气速率和畜禽粪便等农业废弃物的利用率,是当前循环农业领域面临的主要问题。本实验利用自行设计的恒温厌氧发酵装置,模拟农村户用沼气发酵过程,研究不同畜禽粪便混合配比(干物质比)对沼气发酵的影响。厌氧发酵试验表明,单纯发酵原料难以满足产甲烷菌对C:N的需求,分别存在产气启动慢,产气量低等缺点。而通过合理的富氮和富碳的发酵原料配比可以有效地加快发酵产气并提高产气量和沼气中CH4含量。     

秸秆蓝藻混合发酵产沼气过程中微生物群落分析

采用实验室自制秸秆蓝藻混合厌氧反应装置进行沼气发酵实验,利用16S rRNA基因克隆文库的方法,对不同产气阶段的细菌和古菌的优势菌群进行多样性研究。结果表明:(1)不同产气阶段细菌优势种类存在差异,供试秸秆沼气反应器阶段细菌种类较为丰富,分属于6个门:产气初始阶段优势菌群为变形菌门(Proteobacteria),相对丰度为51.76%;产气高峰阶段优势菌群为厚壁菌门(Firmicutes),相对丰度为47.13%;产气结束阶段优势菌群为厚壁菌门(Firmicutes),相对丰度为28.89%;此外,还包括绿弯菌门、螺旋体门、绿菌门的细菌。(2)古菌种类明显少于细菌,均属于甲烷微菌纲(Methanomicrobia)、甲烷杆菌纲(Methanobacteria)和热原体纲(Thermoplasmata)。秸秆蓝藻沼气系统微生物群落结构的阐明具有一定的意义,可为秸秆沼气工程调控提供科学依据。     

Inoculum source for anaerobic fermentation of coffee pulp

Summary An experiment was conducted to find out the correct seeding (inoculum) material for the start up of anaerobic digestion using coffee pulp as substrate. Three different inocula viz., slurry from cowdung biogas plant, sewage sludge and rumen fluid were tried for their efficiency in producing maximum methanogenic activities. The result revealed that the sludge from a biogas plant is the best source of inoculum, which enhanced microbial activities, substrate utilization and gas yield. The various physico-chemical changes that occurred during fermentation were also discussed in detail.     

Bioconversion of wheat stalk to hydrogen by dark fermentation: effect of different mixed microflora on hydrogen yield and cellulose solubilisation

This study determined hydrogen production, volatile fatty acids (VFAs) generation and cellulose solubilisation from anaerobic dark fermentation of wheat stalk and showed the effect of different mixed microflora. The cumulative hydrogen yields of anaerobic digested activated sludge (AS)-inoculated and anaerobic digested dairy manure (DM)-inoculated system were 23.3 and 37.0 mL/g VS at 204 h, respectively. A modified Gompertz equation was able to adequately describe the production of hydrogen from the batch fermentation by both mixed microflora. During the process, acetate and butyrate accounted for more than 76.1% of total VFAs for both fermentations. The extent of cellulose solubilisation approached 46.6% and 75.2% for AS- and DM-inoculated fermentation, respectively. The X-ray diffraction (XRD) showed that the crystallinities of both fermented stalks were partly disrupted by the mixed microflora, and DM-inoculated fermentation had more disruption than AS-inoculated one.     

Psychrophilic methane fermentation of excess sludge by enrichment culture

This study was carried out to determine the possibility of the psychrophilic methane fermentation of the excess sludge produced from fish-processing wastewater treatment. Three different kinds of seed sludge, named M seed sludge, K seed sludge and KL seed sludge were used for batch methane fermentation in Erlenmeyer flasks. M seed sludge was mesophilic, while the other two were prepared in our laboratory by soil collection to obtain a psychrophilic biological community. The effects of temperature on the total volume of biogas generated and on the rate of biogas generation were examined. The activity of K seen sludge with respect to gas yield at 15°C was found to be 1.6-fold higher than that of M seed sludge. Enrichment techniques were then used to accumulate psychrophilic bacteria in KL seed sludge, and the activity at 15°C increased by about two times due to these techniques. Comparison of the three kinds of seed sludge indicates that methane fermentation at low temperature may be feasible, and therefore further study to isolate the psychrophilic bacteria in anaerobic digestion is needed.     

Feasibility of hydrogen production in thermophilic mixed fermentation by natural anaerobes

The biological sludge from an animal wastewater treatment plant was treated to enrich hydrogen-producing mixed bacteria, and effects on hydrogen yield were investigated during anaerobic fermentation at 55 degrees C. Enrichment of hydrogen-producing bacteria was conducted at pH adjustment of inocula to 3 and 5 with and without additional heat treatment (NHT and HT). The enriched mixed bacteria were cultivated at initial pHs of 5, 6, and 7 with synthetic organic wastewater containing different levels of nitrogen (2.0 and 0.8 g/l as total nitrogen) under static batch conditions. The main effects of heat treatment and enrichment pH were significant on hydrogen production. There was no significant effect of different nitrogen concentrations on hydrogen production. The methane-free biogas contained hydrogen levels of up to 64% for a fermentative condition that showed maximum hydrogen evolution (at culture pH 5 after enrichment at pH 5 with HT). The dominating intermediate metabolites were acetate, n-butyrate, and ethanol. Yields of produced hydrogen were significantly dependent upon levels of n-butyrate.     

Hydrogen production with immobilized sewage sludge in three-phase fluidized-bed bioreactors

Municipal sewage sludge was immobilized with a modified alginate gel entrapment method, and the immobilized cells were used to produce hydrogen gas in a three-phase fluidized bed. The hydrogen-producing fluidized beds were operated at different liquid velocity (U(0)) and hydraulic retention time (HRT). The results show that in response to operating liquid velocities, the fluidized-bed system had three flow regimes, namely, plug flow, slug flow, and free bubbling. Pressure fluctuation analysis was used to analyze the hydrodynamic properties in this three-phase fluidized bed when it was under a steady-state production of biogas. With a steady-state biogas production rate (U(g)) of 0.196 mL/s/L, a transition state occurred at a liquid velocity (U(0)) of 0.85 cm/s. As U(0) < 0.85 cm/s, the system was basically a nonhomogeneous fluidized bed, whereas the bed became homogeneous when U(0) was higher than 0.85 cm/s. The fluidized bed can be stably carried out at high loading rates (HRT as low as 2 h). Hydrogen fermentation results show that the maximal hydrogen production rate was 0.93 L/h/L and the best yield (Y(H)2(/sucrose)) was 2.67 mol H(2)/mol sucrose.     

剩余活性污泥完全资源化利用微生物集成技术

剩余活性污泥完全资源化利用微生物集成技术包括: 使用土著PHA合成菌回注法驯化并发酵活性污泥, 生产生物降解材料聚羟基脂肪酸酯(PHA); 采用土著嗜酸性氧化亚铁硫杆菌和氧化硫硫杆菌进行生物淋滤, 去除重金属; 以解磷菌和解钾菌为菌种, 进行固态发酵, 生产生物菌肥。结果表明, 500 L中试PHA占挥发性悬浮固体的20%以上; 重金属含量达到国家排放要求; 生物菌肥中活菌数大于1×108 个/g以上。实现了剩余活性污泥的近零排放。     

Pretreatment of municipal waste activated sludge for volatile sulfur compounds control in anaerobic digestion

The effect of combination of mechanical and chemical pretreatment of municipal waste activated sludge (WAS) prior to anaerobic digestion was studied using a laboratory scale system with an objective to decrease volatile sulfur compounds in biogas and digested sludge. Mechanical pretreatment was conducted using depressurization of WAS through a valve from a batch pretreatment reactor pressurized at 75 ± 1 psi, while combined pretreatments were conducted using six different dosages of hydrogen peroxide (H₂O₂) and ferrous chloride (FeCl₂) along with mechanical pretreatment. About 37-46% removal of H₂S in biogas occurred for different combined pretreatment conditions. Sludge solubilization achieved due to the mechanical pretreatment increased total cumulative methane production by 8-10% after 30 days during the biochemical methane potential (BMP) test. The pretreatment also improved dewaterability in terms of time to filter (TTF), and decreased methyl mercaptan generation potential of the digested sludge.     

微生物烟气脱硫中硫酸盐还原阶段的限制性因素及其影响

【目的】利用硫酸盐还原菌(SRB)厌氧活性污泥进行烟气脱硫,探索硫酸盐生物还原的最适条件及重金属离子对硫酸盐生物还原的影响,以提高硫酸盐还原阶段的效率。【方法】对取自污水处理厂的SRB厌氧活性污泥进行高浓度硫酸盐胁迫驯化。分析生物脱硫过程中SRB厌氧污泥还原硫酸盐的限制性因素及影响。【结果】在最适生长条件下(pH 6.5,32°C),经驯化获得的SRB厌氧活性污泥有较强的硫酸盐还原能力。Fe2+的适量添加对硫酸盐还原有一定促进作用。SRB厌氧污泥还原硫酸盐的ThCOD/SO42-最适值为3.00,ThCOD=3.33为最适理论化学需氧量,硫酸盐还原率可达72.15%。SRB厌氧污泥还原硫酸盐反应体系中抑制SRB活性的硫化物浓度为300 mg/L。Pb2+和Ni2+在较低的浓度下(1.0 mg/L和2.0 mg/L)对硫酸盐的还原产生较强的抑制作用,而Cu2+在稍高的浓度下(8.0 mg/L)显示出明显的抑制作用。【结论】经驯化,SRB厌氧活性污泥显示出较强的硫酸盐还原能力,具有应用于工业烟气生物脱硫的潜力。去除重金属离子Pb2+、Ni2+和Cu2+可有效解除对硫酸盐生物还原作用的抑制。