不同预处理条件对海水养殖废弃物发酵产氢的影响

【目的】利用海水养殖场有机废弃物厌氧发酵产氢,可在减少有机污染物的同时获取氢气。【方法】以海水养殖场有机废弃物为底物,比较嗜热酶(S-TE)、酸、碱、灭菌、微波不同预处理方法对厌氧发酵产氢效果的影响,并对发酵过程中底物性质变化[SCOD、可溶性蛋白质、可溶性糖、pH、VFAs(挥发性脂肪酸)和乙醇]进行探讨。【结果】灭菌预处理产氢效果最好,产氢率为22.0 mL/g VSS,酸处理的效果最差,产氢率为7.6 mL/g VSS。可溶性糖大量消耗之后,氢气不再产生。接种S-TE预处理污泥的底物能更多地释放营养物质,并在整个发酵过程中保持较为稳定的pH值。发酵过程中产生的VFAs主要成分是乙酸,在发酵后期出现乙醇。【结论】灭菌预处理是海水养殖场有机废弃物厌氧发酵产氢的最佳预处理方法,可溶性糖为这一过程主要的营养来源。     

初始pH值对产氢产乙酸/耗氢产乙酸两段耦合工艺定向生产乙酸的影响

以葡萄糖为底物,以经加热预处理并活化过的厌氧污泥为种泥,研究了初始pH值对产氢产乙酸/耗氢产乙酸两段耦合工艺厌氧发酵定向生产乙酸的影响。实验考察了7个初始pH值(5、6、7、8、9、10、11)条件下的底物降解、产物产生和发酵过程pH值的变化。结果表明:产氢产乙酸段初始pH值的变化不仅影响本阶段产酸,而且影响耗氢产乙酸段产酸。初始pH=5时主要进行乙醇型发酵;pH=6和7时主要进行丁酸型发酵;pH=8时混合酸型发酵类型逐渐占优势,pH=8~11时均以乙酸为主要产物,耦合系统生产乙酸最优初始pH值为10。在初始pH=8~11范围内,产氢产乙酸段初期的乙醇浓度一般较高,但到后期因乙醇被微生物进一步代谢转化成乙酸而使其含量下降。     

多级逆流工艺促进城市污泥厌氧发酵生产挥发性脂肪酸

采用一种新型的厌氧发酵工艺——多级逆流发酵工艺对城市污泥进行厌氧发酵, 实现高效产挥发性脂肪酸的目的。结果表明, 实验条件下应用多级逆流发酵工艺, 挥发性脂肪酸浓度与产率分别达到(10.5±0.5) g/L和0.20 gVFAs/ gVS, 与普通厌氧发酵工艺相比, 分别提高了31%和54%。此外, 在多级逆流工艺中, 底物有机质去除率可达50%, 较普通厌氧发酵提高了37%。进一步分析多级逆流工艺产酸的机制, 发现产酸效率的提高在于降低了发酵产物对厌氧产酸细菌的抑制效应, 并且工艺的VFAs产率以及有机质去除率分别取决于第一级和第三级厌氧发酵过程。因此, 城市污泥采用多级逆流工艺厌氧发酵不仅能够有效促进挥发性脂肪酸的生成, 而且能够较大程度上提高污泥中有机质的去除率。     

生物强化污泥厌氧发酵产酸研究进展*

污泥厌氧发酵生产挥发性脂肪酸相较产甲烷,是更具应用价值的污泥稳定途径及资源化利用方式,得到国内外学者的普遍重视。考虑到产酸量低和产酸过程的不稳定性是限制污泥发酵产酸的主要问题,采用生物强化方法实现挥发性脂肪酸的大量积累,与物理和化学方法相比,具有成本低、无二次污染等优点。根据生物强化制剂的类型,归纳了微生物纯培养物、微生物混合培养物及生物酶强化对污泥厌氧发酵产酸的影响,并在此基础上对生物强化技术控制污泥定向产酸、调控奇偶数碳比率等方面的应用进行讨论。此外,分析了影响挥发性脂肪酸产量和组分的因素,如pH、温度、底物、水力停留时间和污泥龄等。最后对生物强化技术的发展方向进行了展望,以期为深入探究污泥资源化利用提供借鉴。     

污泥厌氧消化产酸发酵过程中乙酸累积机制

[目的]研究污泥厌氧消化产挥发性脂肪酸(VFA)过程中的有机物碳流的转化机制,阐明乙酸累积机理。[方法]研究溴乙烷磺酸盐(BES)和氯仿(CHCl3)抑制模型下中间代谢产物和气体的累积,检测各产乙酸功能菌群数量,推断污泥产酸发酵过程中的有机物碳流方向和乙酸累积机理。[结果]BES模型乙酸浓度达27 mmol/L,fhs基因拷贝数比对照组高2-3倍,产氢产乙酸菌略有下降。CHCl3模型乙酸浓度达22 mmol/L,fhs基因拷贝数比BES组低一个数量级,产氢产乙酸菌下降明显。[结论]BES特异性较高,除产甲烷菌外对其他厌氧产酸细菌没有影响,乙酸浓度增加并且其主要来源于水解发酵产酸以及同型产乙酸过程。氯仿除抑制产甲烷菌外,对同型乙酸菌和产氢产乙酸菌也有强烈的抑制作用。     

不同生境微生物转化H2/CO2产乙酸及其在合成气发酵中应用

【目的】合成气发酵对大力开发可再生资源和促进国家可持续发展具有重要意义,研究旨在探究不同生境微生物转化H2/CO2产乙酸及其合成气发酵的潜力。【方法】采集剩余污泥、牛粪、产甲烷污泥和河道底物样品在中温(37 °C)条件下生物转化H2/CO2气体,将来源于牛粪样品的H2/CO2转化富集物用于合成气发酵,通过454高通量技术和定量PCR技术分析复杂微生物群落的组成,GC气相色谱法检测气体转化产生的挥发性脂肪酸(VFAs)浓度。【结果】牛粪和剩余污泥微生物利用H2/CO2气体生成乙酸、乙醇和丁酸等,最高乙酸浓度分别为63 mmol/L和40 mmol/L,明显高于河道底物和产甲烷污泥样品的最高乙酸浓度3 mmol/L和16 mmol/L。牛粪和剩余污泥微生物中含有种类多样化的同型产乙酸菌,剩余污泥中同型产乙酸菌主要为Clostridium spp.、Sporomusa malonica和Acetoanaerobium noterae,牛粪中则为Clostridium spp.、Treponema azotonutricium和Oxobacter pfennigii。【结论】同型产乙酸菌的丰富度和数量两个因素都对复杂微生物群落转化H2/CO2产乙酸效率至关重要;转化H2/CO2得到的富集物可用于合成气发酵产乙酸和乙醇,这为基于混合培养技术的合成气发酵提供了依据。     

不同生境微生物转化H_2/CO_2产乙酸及其在合成气发酵中应用（英文）

【目的】合成气发酵对大力开发可再生资源和促进国家可持续发展具有重要意义,研究旨在探究不同生境微生物转化H_2/CO_2产乙酸及其合成气发酵的潜力。【方法】采集剩余污泥、牛粪、产甲烷污泥和河道底物样品在中温(37°C)条件下生物转化H_2/CO_2气体,将来源于牛粪样品的H_2/CO_2转化富集物用于合成气发酵,通过454高通量技术和定量PCR技术分析复杂微生物群落的组成,GC气相色谱法检测气体转化产生的挥发性脂肪酸(VFAs)浓度。【结果】牛粪和剩余污泥微生物利用H_2/CO_2气体生成乙酸、乙醇和丁酸等,最高乙酸浓度分别为63 mmol/L和40 mmol/L,明显高于河道底物和产甲烷污泥样品的最高乙酸浓度3 mmol/L和16 mmol/L。牛粪和剩余污泥微生物中含有种类多样化的同型产乙酸菌,剩余污泥中同型产乙酸菌主要为Clostridium spp.、Sporomusa malonica和Acetoanaerobium noterae,牛粪中则为Clostridium spp.、Treponema azotonutricium和Oxobacter pfennigii。【结论】同型产乙酸菌的丰富度和数量两个因素都对复杂微生物群落转化H_2/CO_2产乙酸效率至关重要;转化H_2/CO_2得到的富集物可用于合成气发酵产乙酸和乙醇,这为基于混合培养技术的合成气发酵提供了依据。     

玉米芯发酵法生物制氢

在批式培养试验中, 以牛粪堆肥为天然产氢菌源, 玉米芯为底物, 通过厌氧发酵生产氢气。系统考察了底物预处理条件、初始pH值和底物浓度对玉米芯产氢能力的影响。在初始pH 8.0, 1.0%盐酸预处理底物30 min, 底物浓度10 g/L的最佳产氢条件下, 玉米芯最大产氢能力〔每克TVS（总挥发性固体物）产氢量〕和最大产氢速率（每克TVS每小时产氢量）分别为107.9 mL /g、4.20 mL/g·h-1。玉米芯经酸预处理后半纤维素含量由42.2%下降至3.0%, 而酸预处理的玉米芯产氢前后纤维素、半纤维素和木质素含量只有少量变化。产氢菌主要用酸预处理产生的可溶性糖产氢, 故底物的酸预处理对玉米芯的发酵产氢非常重要。用傅里叶变换红外光谱(FTIR)分析显示酸预处理和产氢过程中玉米芯的特征峰发生变化, 酸预处理过程降解了底物纤维素的无定形区和半纤维素, 产氢微生物对纤维素的结晶区有破坏作用。     

玉米芯发酵法生物制氢

在批式培养试验中, 以牛粪堆肥为天然产氢菌源, 玉米芯为底物, 通过厌氧发酵生产氢气。系统考察了底物预处理条件、初始pH值和底物浓度对玉米芯产氢能力的影响。在初始pH 8.0, 1.0%盐酸预处理底物30 min, 底物浓度10 g/L的最佳产氢条件下, 玉米芯最大产氢能力〔每克TVS（总挥发性固体物）产氢量〕和最大产氢速率（每克TVS每小时产氢量）分别为107.9 mL /g、4.20 mL/g·h-1。玉米芯经酸预处理后半纤维素含量由42.2%下降至3.0%, 而酸预处理的玉米芯产氢前后纤维素、半纤维素和木质素含量只有少量变化。产氢菌主要用酸预处理产生的可溶性糖产氢, 故底物的酸预处理对玉米芯的发酵产氢非常重要。用傅里叶变换红外光谱(FTIR)分析显示酸预处理和产氢过程中玉米芯的特征峰发生变化, 酸预处理过程降解了底物纤维素的无定形区和半纤维素, 产氢微生物对纤维素的结晶区有破坏作用。     

污泥厌氧消化中同型产乙酸菌富集研究

在市政污泥厌氧消化过程中,采用三阶段选择性富集同型产乙酸菌,通过监测培养过程中pH值、挥发性脂肪酸、关键酶的活性以及不同碳源利用率等,研究同型产乙酸菌在系统中的累积情况.实验数据表明:经过40 d的富集培养,pH值稳定在8.0,乙酸含量逐渐趋于稳定,占COD的比例为46.4％,同型产乙酸菌对底物的利用率达到了87％,乙酸激酶和乙酰磷酸转移酶呈现明显上升的趋势.结果显示,通过本方法可在市政污泥中有效地富集同型产乙酸菌.     

Investigation of an acetate-fed denitrifying microbial community by stable isotope probing, full-cycle rRNA analysis, and fluorescent in situ hybridization-microautoradiography

The acetate-utilizing microbial consortium in a full-scale activated sludge process was investigated without prior enrichment using stable isotope probing (SIP). [13C]acetate was used in SIP to label the DNA of the denitrifiers. The [13C]DNA fraction that was extracted was subjected to a full-cycle rRNA analysis. The dominant 16S rRNA gene phylotypes in the 13C library were closely related to the bacterial families Comamonadaceae and Rhodocyclaceae in the class Betaproteobacteria. Seven oligonucleotide probes for use in fluorescent in situ hybridization (FISH) were designed to specifically target these clones. Application of these probes to the sludge of a continuously fed denitrifying sequencing batch reactor (CFDSBR) operated for 16 days revealed that there was a significant positive correlation between the CFDSBR denitrification rate and the relative abundance of all probe-targeted bacteria in the CFDSBR community. FISH-microautoradiography demonstrated that the DEN581 and DEN124 probe-targeted cells that dominated the CFDSBR were capable of taking up [14C]acetate under anoxic conditions. Initially, DEN444 and DEN1454 probe-targeted bacteria also dominated the CFDSBR biomass, but eventually DEN581 and DEN124 probe-targeted bacteria were the dominant bacterial groups. All probe-targeted bacteria assessed in this study were denitrifiers capable of utilizing acetate as a source of carbon. The rapid increase in the number of organisms positively correlated with the immediate increase in denitrification rates observed by plant operators when acetate is used as an external source of carbon to enhance denitrification. We suggest that the impact of bacteria on activated sludge subjected to intermittent acetate supplementation should be assessed prior to the widespread use of acetate in the wastewater industry to enhance denitrification.     

Dynamic transition of a methanogenic population in response to the concentration of volatile fatty acids in a thermophilic anaerobic digester

In this study, the microbial community succession in a thermophilic methanogenic bioreactor under deteriorative and stable conditions that were induced by acidification and neutralization, respectively, was investigated using PCR-mediated single-strand conformation polymorphism (SSCP) based on the 16S rRNA gene, quantitative PCR, and fluorescence in situ hybridization (FISH). The SSCP analysis indicated that the archaeal community structure was closely correlated with the volatile fatty acid (VFA) concentration, while the bacterial population was impacted by pH. The archaeal community consisted mainly of two species of hydrogenotrophic methanogen (i.e., a Methanoculleus sp. and a Methanothermobacter sp.) and one species of aceticlastic methanogen (i.e., a Methanosarcina sp.). The quantitative PCR of the 16S rRNA gene from each methanogen revealed that the Methanoculleus sp. predominated among the methanogens during operation under stable conditions in the absence of VFAs. Accumulation of VFAs induced a dynamic transition of hydrogenotrophic methanogens, and in particular, a drastic change (i.e., an approximately 10,000-fold increase) in the amount of the 16S rRNA gene from the Methanothermobacter sp. The predominance of the one species of hydrogenotrophic methanogen was replaced by that of the other in response to the VFA concentration, suggesting that the dissolved hydrogen concentration played a decisive role in the predominance. The hydrogenotrophic methanogens existed close to bacteria in aggregates, and a transition of the associated bacteria was also observed by FISH analyses. The degradation of acetate accumulated during operation under deteriorative conditions was concomitant with the selective proliferation of the Methanosarcina sp., indicating effective acetate degradation by the aceticlastic methanogen. The simple methanogenic population in the thermophilic anaerobic digester significantly responded to the environmental conditions, especially to the concentration of VFAs.     

Dynamic Transition of a Methanogenic Population in Response to the Concentration of Volatile Fatty Acids in a Thermophilic Anaerobic Digester

In this study, the microbial community succession in a thermophilic methanogenic bioreactor under deteriorative and stable conditions that were induced by acidification and neutralization, respectively, was investigated using PCR-mediated single-strand conformation polymorphism (SSCP) based on the 16S rRNA gene, quantitative PCR, and fluorescence in situ hybridization (FISH). The SSCP analysis indicated that the archaeal community structure was closely correlated with the volatile fatty acid (VFA) concentration, while the bacterial population was impacted by pH. The archaeal community consisted mainly of two species of hydrogenotrophic methanogen (i.e., a Methanoculleus sp. and a Methanothermobacter sp.) and one species of aceticlastic methanogen (i.e., a Methanosarcina sp.). The quantitative PCR of the 16S rRNA gene from each methanogen revealed that the Methanoculleus sp. predominated among the methanogens during operation under stable conditions in the absence of VFAs. Accumulation of VFAs induced a dynamic transition of hydrogenotrophic methanogens, and in particular, a drastic change (i.e., an approximately 10,000-fold increase) in the amount of the 16S rRNA gene from the Methanothermobacter sp. The predominance of the one species of hydrogenotrophic methanogen was replaced by that of the other in response to the VFA concentration, suggesting that the dissolved hydrogen concentration played a decisive role in the predominance. The hydrogenotrophic methanogens existed close to bacteria in aggregates, and a transition of the associated bacteria was also observed by FISH analyses. The degradation of acetate accumulated during operation under deteriorative conditions was concomitant with the selective proliferation of the Methanosarcina sp., indicating effective acetate degradation by the aceticlastic methanogen. The simple methanogenic population in the thermophilic anaerobic digester significantly responded to the environmental conditions, especially to the concentration of VFAs.     

一株新型同型产乙酸菌Clostridium sp.的自养和异养生长特性

【背景】同型产乙酸菌是一类利用乙酰辅酶A途径固定CO_2合成自身细胞物质并生成乙酸、乙醇等代谢产物的厌氧菌群,其分布广泛、种类繁多且代谢多样。深入研究同型产乙酸菌菌株的代谢能力及特性,对探索该种群的生理生化特性及其环境作用至关重要。【目的】研究一株同型产乙酸菌Clostridium sp. BXX的最适培养条件及其自养与异养生长特性。【方法】设置BXX菌株培养温度10-55°C、初始pH 6.0-9.0、NaCl浓度0-2.0%、不同氮源,测定菌体细胞含量和产物生成浓度,确定菌株最适培养条件。研究BXX菌株分别以H_2/CO_2、合成气、CO、葡萄糖、1,2-丙二醇、甲酸钠、乙二醇甲醚、甘油、丙酮酸和乳酸为底物时的底物消耗、产物生成、菌体细胞含量和pH等,探究其自养和异养生长特性。【结果】BXX菌株的最适培养温度为30°C,初始pH为7.0,NaCl浓度为1.0%,氮源为酵母粉。BXX菌株能以H2/CO2、合成气、葡萄糖、1,2-丙二醇、甲酸钠、乙二醇甲醚和甘油为底物生长,不能以CO、丙酮酸或乳酸为底物生长。【结论】BXX菌株既能自养生长产乙酸,又能异养生长产乙醇。BXX菌株是乙酸发酵的优良菌种资源,有较好的工业应用潜力。     

Response surface methodology analysis of anaerobic syntrophic degradation of volatile fatty acids in an upflow anaerobic sludge bed reactor inoculated with enriched cultures

Anaerobic oxidation of volatile fatty acids (VFAs) as the key intermediates is restricted thermodynamically. Presently, enriched acetogenic and methanogenic cultures were used for syntrophic anaerobic digestion of VFAs in an upflow anaerobic sludge bed reactor fed with acetic, propionic, and butyric acids at maximum concentrations of 5.0, 3.0, and 4.0 g/L, respectively. Interactive effects of propionate, butyrate and acetate were analyzed. Hydraulic retention time (HRT) and acetate oxidizing syntrophs and methanogen (hydrogenotrophs) to syntrophic bacteria (propionate- and butyrate-oxidizing bacteria) population ratio (M/A) were investigated as key microbiological and operating variables of VFA anaerobic degradations. M/A did not affect the size distribution and had little effect on extracellular polymer contents of the granules. Granular sludge with close spatial microbial proximity enhanced syntrophic degradation of VFAs compared to other cultures, such as suspended cultures. Optimum conditions were found to be propionate = 1.93 g/L, butyrate = 2.15 g/L, acetate = 2.50 g/L, HRT = 22 h, and M/A = 2.5 corresponding to maximum VFA removal and biogas production rate. Results of verification experiments and predicted values from fitted correlations were in close agreement at the 95% confidence interval. Granules seemed to be smaller particles and less stable in construction with an irregular fractured surface compared to the original granules.     

pH值和产甲烷抑制剂对两相耦合系统污泥发酵定向产乙酸的影响

采用产氢产乙酸/同型产乙酸两相耦合工艺对剩余污泥进行了半连续式厌氧发酵,主要研究了pH值和产甲烷抑制剂2-bromoethanesulphonate(BES)对耦合系统定向产乙酸的影响.结果表明:碱性pH(pH=10.0)和添加BES都能促进A相乙酸的积累,提高乙酸的产率,同时碱性pH比添加BES更有利于污泥的水解.当...     

Production of polyhydroxyalkanoates by activated sludge treating a paper mill wastewater

Production of polyhydroxyalkanoates (PHAs) in activated sludge treating wastewater represents an economical and environmental promising alternative to pure culture fermentations. A process for production of PHA from a paper mill wastewater was examined at laboratory scale. The three stage process examined consisted of acidogenic fermentation to convert wastewater organic matter to volatile fatty acids (VFAs), an activated sludge system operating under feast/famine conditions to enrich for PHA producing organisms and accumulation of PHA in batch experiments. After fermentation of the wastewater, 74% of the soluble COD was present as VFA (acetate, propionate, butyrate and valerate) and the resulting PHA after batch accumulation consisted of 31-47 mol% hydroxybutyrate and 53-69 mol% hydroxyvalerate. The maximum PHA content achieved was 48% of the sludge dry weight and the three stage process exhibited a potential to produce 0.11 kg of PHA per kg of influent COD treated.     

Molecular analysis of halophilic bacterial community for high-rate denitrification of saline industrial wastewater

A denitrification system for saline wastewater utilizing halophilic denitrifying bacteria has not been developed so far. In this study, denitrification performance and microbial community under various saline conditions were investigated using denitrifying sludge acclimated under low-salinity condition for a few years as seed sludge. A continuous denitrification experiment showed that denitrification performance and microbial community at 10% salinity was higher than that at 1% salinity. The microbial community in the denitrification sludge that was acclimated under low salinity was monitored by terminal-restriction fragment length polymorphism (T-RFLP) analysis during acclimation to high-salinity condition. T-RFLP profiles and clone analysis based on 16S rRNA-encoding genes in the sludge of the denitrification system with 10% salinity indicated that the γ-Proteobacteria, particularly Halomonas spp., were predominant species, suggesting that these bacterial members were possibly responsible for a high denitrification activity under high-salinity conditions. Furthermore, the investigation of denitrification performance under various saline conditions revealed that 4–10% salinity results in the highest denitrification rate, indicating that this salinity was optimal for predominant bacterial species to exhibit denitrification activity. These results indicate the possibility that an appropriate denitrification system for saline wastewater can be designed using acclimated sludge with a halophilic community.     

Combination of fluorescent in situ hybridization and microautoradiography-a new tool for structure-function analyses in microbial ecology

A new microscopic method for simultaneously determining in situ the identities, activities, and specific substrate uptake profiles of individual bacterial cells within complex microbial communities was developed by combining fluorescent in situ hybridization (FISH) performed with rRNA-targeted oligonucleotide probes and microautoradiography. This method was evaluated by using defined artificial mixtures of Escherichia coli and Herpetosiphon aurantiacus under aerobic incubation conditions with added [3H]glucose. Subsequently, we were able to demonstrate the potential of this method by visualizing the uptake of organic and inorganic radiolabeled substrates ([14C]acetate, [14C]butyrate, [14C]bicarbonate, and 33Pi) in probe-defined populations from complex activated sludge microbial communities by using aerobic incubation conditions and anaerobic incubation conditions (with and without nitrate). For both defined cell mixtures and activated sludge, the method proved to be useful for simultaneous identification and analysis of the uptake of labeled substrates under the different experimental conditions used. Optimal results were obtained when fluorescently labeled oligonucleotides were applied prior to the microautoradiographic developing procedure. For single-cell resolution of FISH and microautoradiographic signals within activated sludge flocs, cryosectioned sample material was examined with a confocal laser scanning microscope. The combination of in situ rRNA hybridization techniques, cryosectioning, microautoradiography, and confocal laser scanning microscopy provides a unique opportunity for obtaining cultivation-independent insights into the structure and function of bacterial communities.     

High-rate anaerobic treatment of wastewater at low temperatures

Anaerobic treatment of a volatile fatty acid (VFA) mixture was investigated under psychrophilic (3 to 8 degrees C) conditions in two laboratory-scale expanded granular sludge bed reactor stages in series. The reactor system was seeded with mesophilic methanogenic granular sludge and fed with a mixture of VFAs. Good removal of fatty acids was achieved in the two-stage system. Relative high levels of propionate were present in the effluent of the first stage, but propionate was efficiently removed in the second stage, where a low hydrogen partial pressure and a low acetate concentration were advantageous for propionate oxidation. The specific VFA-degrading activities of the sludge in each of the modules doubled during system operation for 150 days, indicating a good enrichment of methanogens and proton-reducing acetogenic bacteria at such low temperatures. The specific degradation rates of butyrate, propionate, and the VFA mixture amounted to 0.139, 0.110, and 0.214 g of chemical oxygen demand g of volatile suspended solids-1 day-1, respectively. The biomass which was obtained after 1.5 years still had a temperature optimum of between 30 and 40 degrees C.