生物淋滤法去除污泥中的重金属

本研究从剩余活性污泥中分离得到两株土著硫杆菌。对两株菌进行了分类鉴定。确立二者分别为嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans, A. f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans, A. t)。将二者的单菌和混合菌分别接种于剩余活性污泥中, 进行了为期9 d的生物淋滤, 对淋滤过程中的pH变化、氧化还原电位(ORP)以及重金属含量进行了检测。结果表明, 生物淋滤9 d混合菌对于As、Cr、Cu、Ni和Zn的去除效果最好; 去除率分别达到了96.09%、93.47%、98.32%、97.88%和98.60%。对于Cd和Pb混合菌生物淋滤的去除率在第6天之后迅速下降, 但是A. t单菌淋滤保持较高的去除率。     

污泥中重金属的生物淋滤

生物淋滤法(Bioleaching)是指利用自然界中一些微生物(硫细菌)的直接作用或其代谢产物的间接作用,产生氧化、还原、络合、吸附或溶解反应,将固相中某些不溶性成分(如重金属、硫及其他金属)分离浸提出来的技术.在生物淋滤中,嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,A.f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans,A.f)被用作有效的淋滤载体[1].这种嗜酸性的化能自养型细菌以大气中的CO2为碳源,以无机物铁或硫为能源来维持生长,不需要提供外来的碳源和电子供体.另外,由于pH值很低,抑制了其他细菌的生长,所以在实际的操作过程中不需要严格的无菌条件.氧化亚铁硫杆菌和氧化硫硫杆菌去除重金属适宜于污水处理厂的开放系统,采用土著嗜酸性氧化亚铁硫杆菌(A.f)和氧化硫硫杆菌(A.f)进行重金属去除.也就是说,处理什么地方的污泥,就在什么地方分离A.f和A.t,这样分离的微生物在生物淋滤过程中能发挥较好的作用.这也是微生物在自然界生长繁殖的特点之一. 本期介绍了王聪、宋存江等[2]从剩余活性污泥中分离得到两株土著硫杆菌,对两株菌进行了分类鉴定,确定二者分别为嗜酸性氧化亚铁硫杆菌杆(Acidithiobacillus ferrooxidans,A.f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans,A.t),将二者的单菌和混合菌分别接种于剩余活性污泥中,进行了为期9 d的生物淋滤,对淋滤过程中的pH变化、氧化还原电位(ORP)以及重金属含量进行了检测.结果表明,生物淋滤9 d的混合菌对于As、Cr、Cu、Ni和Zn的去除效果最好,去除率分别达到了96.09%、93.47%、98.32%、97.88%和98.60%.混合菌生物淋滤对于Cd和Pb的去除率在第6天之后迅速下降,但是A.t单菌淋滤保持较高的去除率,此结果为进一步的应用打下了良好的基础.     

门多萨假单胞菌NK-01合成中长链聚羟基脂肪酸酯的发酵条件优化

为了提高PHA_MCL在门多萨假单胞菌NK-01中的积累,采用单因素实验和正交实验确立了发酵生产PHA_MCL的最佳条件,即以PHA产量为指标的最佳发酵条件为15 g/L葡萄糖浓度、C/N=50、发酵时间48 h,该条件下获得产量0.8 g/L以上的PHA;以PHA占菌体干重百分含量为指标的最佳发酵条件为10 g/L葡萄糖浓度、C/N=60、发酵时间48 h,该条件下获得占菌体干重50%以上的PHA。该研究将为门多萨假单胞菌NK-01用于PHA_MCL的规模化生产提供理论依据。     

厌氧发酵液中溶解性有机物对活性污泥合成聚羟基脂肪酸酯影响的研究进展

利用活性污泥微生物将剩余污泥发酵液中的挥发性脂肪酸(Volatile fatty acids,VFAs)转化为聚羟基脂肪酸酯(Polyhydroxyalkanoates,PHA)是目前环境生物技术领域的研究热点。但针对发酵液中非VFAs物质(主要是溶解性有机物,Dissolved organic matter,DOM)对活性污泥合成PHA的影响目前仍未有统一的结论,需要进行较全面的分析和总结。因此,文中首先介绍了剩余污泥发酵液中DOM的主要特性及常见分析方法,然后从微生物学、代谢调控、污泥性质等角度分析了DOM中几种主要组分(多聚糖、蛋白类物质、腐殖质)对活性污泥合成PHA的影响。综合各研究结果表明高浓度DOM不利于PHA的生产,但适量DOM的存在有利于污泥性质的稳定,同时能降低PHA提纯成本。最后提出了相应的策略来实现对活性污泥利用DOM合成PHA的调控,以期为高效利用厌氧发酵液中DOM实现PHA合成提供解决思路。     

污水处理过程中微生物群落多样性及其对环境因子响应的研究进展

污水生物处理系统的性能和稳定性与微生物群落结构和动态密切相关。通过深入了解活性污泥中微生物群落结构及其影响因素,有助于提高污水厂污染物的去除效果。在不同污水活性污泥处理系统中细菌群落主要以变形菌、绿弯菌、放线菌、厚壁菌和拟杆菌为功能菌群;活性污泥中寄居的大多数真菌来自于子囊菌门,还有少量担子菌门;古菌以产甲烷菌为主;而病毒中分布最广的噬菌体和致病性病毒是最主要的关注点。本文通过对相关文献分析及总结,综述了进水组成、不同处理工艺、参数(理化参数和运行参数)、地理位置和气候条件等环境因子对活性污泥中细菌、真菌、古菌以及病毒群落组成的影响,尽可能全面地介绍污水厂微生物群落多样性及其对环境因子的响应。同时,对未来研究方向进行探讨,以期能够为活性污泥中功能微生物的应用及调控提供理论和应用基础。     

活性污泥中菌群多样性及其功能调控研究进展

活性污泥是污水处理厂生物处理工艺的功能主体,活性污泥中菌群的种类、数量及活性是提高污水处理能力与效果的重要基础。本文综述了活性污泥处理工艺中的主要功能细菌(絮凝菌、脱氮菌、除磷菌等)生物群落的多样性与生态特征,并对目前主流的菌群鉴定方式进行总结,最后从运行条件、定向驯化及生物强化3个方面对菌群调控进行论述,以期为活性污泥法污水处理工艺提供一些理论指导。     

活性污泥法生产聚羟基烷酸（PHA）

介绍了厌氧-好氧活性污泥法生产生物降解塑料PHA的生化机制及增加活性污泥中PHA含量的新方法.     

利用剩余活性污泥合成聚羟基脂肪酸酯的研究进展

利用单一微生物发酵是现阶段获得聚羟基脂肪酸酯 (PHA) 的主要方式,但过高的生产成本限制了其大规模应用。近年来利用活性污泥菌群混合培养合成PHA被广泛研究。将剩余污泥处理与PHA合成相结合,不仅可以省去纯培养所必需的灭菌环节,同时可以实现剩余污泥的资源化利用。剩余污泥的水解酸化、菌群富集驯化及PHA合成受环境因素影响,深入的生物合成机制研究有助于混合培养合成PHA的推广应用。文中主要介绍利用剩余污泥合成PHA的可行性、影响剩余污泥水解酸化的因素、污泥菌群富集驯化合成PHA及其机制等方面的研究进展。     

构建伴有β-葡萄糖苷酶表达的乙醇发酵大肠杆菌

研究构建能够分泌表达纤维素酶的产乙醇菌株,实现降解木质纤维素生产乙醇的整合生物加工过程。文中通过克隆来自运动发酵单胞菌Zymomonas mobilis ZM4的丙酮酸脱羧酶基因pdc和乙醇脱氢酶基因adhB,并通过Red重组将二者整合到大肠杆菌Escherichia coli JM109基因组中,首先构建了一株可以利用葡萄糖进行乙醇发酵的重组菌E. coli P81。随后将来源于多粘芽胞杆菌Bacillus polymyxa1.794的β-葡萄糖苷酶基因bglB在E. coli P81中进行了分泌表达,得到了一株可以进行纤维二糖降解和乙醇发酵双重功能的重组菌E. coli P81(pUC19-bglB)。该菌胞外分泌β-糖苷酶活达到84.78 mU/mL菌液,纤维二糖酶活达到了32.32 mU/mL菌液。该重组菌E. coli P81(pUC19-bglB) 以纤维二糖为碳源进行乙醇发酵,乙醇得率达到了理论产率55.8％,而在葡萄糖和纤维二糖的共发酵中,其乙醇产量达到了理论产率46.5％。构建得到的此株整合生物加工大肠杆菌能够利用β-葡萄糖苷酶生产乙醇,为构建能利用木质纤维素分解产物生产燃料乙醇的高效、稳定生产用工程菌奠定了良好的基础。     

VC三菌种一步发酵方法的构建与研究

就维生素C微生物一步发酵方法进行了探索,构建了酮古龙酸杆菌、氧化葡萄糖酸杆菌和芽孢杆菌三菌混菌一步发酵的方法。研究发现,植物内生芽孢杆菌可以与酮古龙酸杆菌配合,促进酮古龙酸杆菌生长和产酸。在有山梨醇存在的条件下酮古龙酸杆菌及其伴生菌能够快速地生长增殖,植物内生芽孢杆菌在发酵的10h中不断消耗山梨醇。5L的发酵罐中,酮古龙酸杆菌、氧化葡萄糖酸杆菌和植物内生芽孢杆菌三菌混菌一步发酵在恒定的30℃温度,600r/min搅拌速度和1.5vvm通气条件下,补料发酵过程中醇酸质量转化率达到了81.89%,在分批发酵过程中,醇酸质量转化率达到了87.90%,进一步优化了维生素C生产工艺。     

Optimal production of polyhydroxyalkanoates (PHA) in activated sludge fed by volatile fatty acids (VFAs) generated from alkaline excess sludge fermentation

To reduce the production cost of polyhydroxyalkanoates (PHA) and disposal amount of excess sludge simultaneously, the feasibility of using fermentative volatile fatty acids (VFAs) as carbon sources to synthesize PHA by activated sludge was examined. At pH 11.0, 60 degrees C and fermentative time of 7d, the VFAs yield was 258.65 mgTOC/gVSS. To restrain cell growth during PHA production, the released phosphorus and residual ammonium in the fermentative VFAs was recovered by the formation of struvite precipitation. Acetic acid was the predominant composition of the fermentative VFAs. PHA accumulation in excess sludge occurred feeding by fermentative VFAs with aerobic dynamic feeding process. The maximum PHA content accounted for 56.5% of the dry cell. It can be concluded from this study that the VFAs generated from excess sludge fermentation were a suitable carbon source for PHA production by activated sludge.     

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.     

微生物接种密度和矿物收集时间对生物沥浸中次生铁矿物形成的影响

在嗜酸性氧化亚铁硫杆菌Acidithiobacillus ferrooxidans作用下, 污泥生物沥浸体系中常会有次生铁矿物形成, 这些矿物对污泥脱水和重金属溶出有重要影响。在FeSO4-K2SO4-H2O生物成矾临界点模拟生物沥浸过程, 考察了Acidithiobacillus ferrooxidans菌接种密度和矿物收集时间对次生铁矿物的影响。结果表明, 微生物接种密度和矿物收集时间对生物沥浸过程中次生铁矿物的重量及其类型均有一定影响, 随矿物收集时间的推迟, 溶液中含有的一价阳离子(如K+等)可导致施氏矿物向黄铁矾发生转变, 并成为影响铁矿物类型的主导因素。     

分离自活性污泥的硫酸盐还原菌用于铅锌冶炼渣重金属污染修复

【背景】从活性污泥中分离出一类具有硫酸盐还原能力的菌株,探讨了其用于铅锌冶炼渣重金属污染修复的可行性。【目的】研究硫酸盐还原菌(Sulfate reducing bacteria)对铅锌冶炼渣中重金属的固化作用。【方法】将从活性污泥中分离出的硫酸盐还原菌接种到铅锌冶炼渣中进行修复,采用X射线衍射、Tessier、电感耦合等离子体发射光谱仪检测、高通量测序等方法进行实验。检测铅锌冶炼渣中矿物组成,以及修复过程中重金属化学形态、各金属离子浓度和微生物群落结构的变化。【结果】修复实验表明,体系中电位降低、pH值提高、各重金属稳定态增加、离子浓度降低且微生物群落结构变化显著,硫酸盐还原菌成为优势菌群。【结论】接种硫酸盐还原菌后铅锌冶炼渣中的重金属原位固化效果显著,从而降低生物可利用性,将恶性循环变为良性循环,所以硫酸盐还原菌可用作重金属污染修复的固化药剂。     

PHA production by activated sludge.

The production of polyhydroxyalkanoate by anaerobic-aerobic activated sludge was reviewed concentrating on the biochemical mechanisms and on the trials to increase polyhydroxyalkanoate (PHA) content in activated sludge. The anaerobic aerobic activated sludge system selects microorganisms with the capabilities to couple glycolysis, polyphosphate degradation, and PHA accumulation for anaerobic substrate uptake. Some of the PHA-related metabolisms observed there have not been seen in pure cultures so far. Such metabolisms are the formation of PHA containing 3-hydroxy-2-methylvalerate, and '3-hydroxyvalerate fermentation' in which glucose or glycogen is converted to 3-hydroxyvalerate-rich PHA while yielding energy. The PHA content of activated sludge can be increased up to 62% by applying a microaerophilic-aerobic activated sludge process. PHA production by activated sludge is worth investigation.     

Detection of polyhydroxyalkanoate-accumulating bacteria from domestic wastewater treatment plant using highly sensitive PCR primers

Polyhydroxyalkanoate (PHA) is a class of biodegradable plastics that have great potential applications in the near future. In this study, the micro-biodiversity and productivity of PHA-accumulating bacteria in activated sludge from a domestic wastewater treatment plant were investigated. A previously reported primer set and a selfdesigned primer set (phaCF1BO/phaCR2BO) were both used to amplify the PHA synthase (phaC) gene of isolated colonies. The new primers demonstrated higher sensitivity for phaC, and combining the PCR results of the two primer sets was able to widen the range of detected genera and raise the sensitivity to nearly 90%. Results showed that 85.3% of the identified bacteria were Gram-negative, with Ralstonia as the dominant genus, and 14.7% were Gram-positive. In addition, Zoogloea and Rhizobium contained the highest amounts of intracellular PHA. It is apparent that glucose was a better carbon source than pentone or tryptone for promoting PHA production in Micrococcus. Two different classes, class I and class II, of phaC were detected from alphaproteobacteria, betaproteobacteria, and gammaproteobacteria, indicating the wide diversity of PHA-accumulating bacteria in this particular sampling site. Simultaneous wastewater treatment and PHA production is promising by adopting the high PHAaccumulating bacteria isolated from activated sludge.     

Acidophilic microbial communities catalyzing sludge bioleaching monitored by fluorescent in situ hybridization

Biological autotrophic sulfur oxidation processes have been proposed to remove heavy metals from wastewater treatment sludge by bioleaching. We made a characterization of the microbial population in batch and continuous sludge bioleaching reactors using fluorescent in situ hybridization of fluorescently-labeled oligonucleotidic probes targeting rRNA in a ‚top to bottom approach’. Batch incubations of sludge with 0.2% (w/v) elemental sulfur resulted in a pH value of 5. Alpha-Proteobacteria hybridizing with probe ALF1b were dominant in this incubation. Members of the Acidophilium-group (hybridizing with probe Acdp821) of Nitrospira/Leptospirillum phylum (Ntspa712 probe) and from the archaeal domain (ARCH915) were also detected. When sludge was incubated with 1% elemental sulfur in batch or continuous reactor experiments, final pH values were always below 2. Active microbial communities consisted almost exclusively of gamma-Proteobacteria (hybridizing with probe GAM42a). However, further hybridization experiments with probe Thio820 targeting Acidithiobacillus ferroxidans and Acidithiobacillus thioxidans gave negative results. A new probe, named THIO181, encompassing all known members of the genus was designed. Hybridization perfomed with THIO181 and GAM42a showed a perfect co-localization of the hybridization signals. Further hybridization experiments with probe THIO181 and THC642, specific for the species Acidithiobacillus caldus, confirmed that this bacteria was largely responsible for the sulfur oxidation reaction in our acidophilic sludge bioleaching reactors.