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.     

嗜酸氧化亚铁硫杆菌基因组学研究进展

嗜酸氧化亚铁硫杆菌(Acidithiobacillus ferrooxidan,A.ferrooxidans)广泛存在于酸性矿物废水中,与生物冶金和环境净化紧密相关。不同来源嗜酸氧化亚铁硫杆菌全基因组的测序,为我们利用比较基因组学和功能基因组学的方法去洞察嗜酸氧化亚铁硫杆菌功能基因,提供了坚实的研究基础和丰富的科研信息。简述了嗜酸氧化亚铁硫杆菌基因组学的基本特征;从比较基因组学和功能基因组学发现了嗜酸氧化亚铁硫杆菌菌株基因组水平的差异;通过生物信息学概述了该菌的铁和硫代谢机制,并从细菌的功能基因组学对其在生物冶金与环境治理等应用进行了展望。     

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。     

一种浸矿混合菌种的筛选、鉴定及浸矿的研究

旨在从某低品位硫化铜矿酸性矿坑水中分离获得浸矿效果优良的细菌,同时对细菌的浸矿行为进行初步的研究。采集福建某低品位硫化铜矿酸性矿坑水样品,9K培养基富集获得混合菌种FIM-201304。利用9K固体平板,从混合菌FIM-201304中分离获得优势浸矿菌D-1。应用高通量Illumina Miseq测序技术分析混合菌FIM-201304的群落结构。通过表型特征和16S rRNA基因序列分析鉴定菌株D-1。采用摇瓶培养对比研究混合菌和单菌对低品位硫化铜矿和低品位硫化镍矿的浸出效果。结果显示,高通量测序技术分析结果表明,混合菌FIM-201304的优势菌是嗜酸氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,A.f errooxidans)(丰度占比85.02%)、铜绿色假单胞菌(Pseudomonas sp.)(丰度占比10.07%)、嗜酸异养菌(Acidiphilium acidophilum,A.acidophilum)(丰度占比1.30%)和鞘氨醇单胞菌(Sphingomonas sp.)(丰度占比1.21%)。表型特征和16S r RNA基因序列分析确定菌株D-1属于氧化亚铁硫杆菌。摇瓶实验结果表明,浸出反应20 d后,接种混合菌的浸出体系中铜、镍浸出率分别为65%和56%,相同条件下,接种单菌的浸出体系中铜、镍的浸出率为41%和36%,接种混合菌的浸出体系比接种单菌的浸出体系中铜、镍浸出率分别提高了24%和20%。混合菌对矿石的浸出效果显著优于单菌。异养菌促进了自养菌对矿石中金属元素的浸出。     

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。     

不同能源条件下中度嗜热嗜酸细菌多样性分析

采用黄铁矿、黄铜矿、硫酸亚铁和硫粉混合物作为主要能源物质在50°C条件下分别培养中度嗜热细菌混合物,研究其细菌多样性。提取细菌基因组总DNA,采用PCR结合限制性酶切片段多态性分析(RFLP)方法进行细菌16SrRNA基因的系统发育分析,比较不同能源条件下富集培养的混合细菌群落构成的差异。从3个培养物中共获得阳性克隆303个并进行RFLP分析,对29种不同酶切谱型的克隆插入序列进行测定和系统发育分析。大部分序列与已报道的浸矿微生物16SrRNA序列相似性较高(89.1%~99.7%),归属于硫化叶菌属的耐温氧化硫化杆菌(Sulfobacillus thermotolerans)和热氧化硫化杆菌(Sulfobacillus thermosulfidooxidans),嗜酸硫杆菌属的喜温硫杆菌(Acidithiobacillus caldus),钩端螺旋菌属的嗜铁钩端螺旋菌(Leptospirillumferriphilum)以及unculturedforest soil bacterium、uncultured proteobacterium。其中Acidithiobacillus caldus,Sulfobacillus thermotolerans,Leptospirillumferriphilum3种细菌为三类能源物质培养物中的优势细菌类群。L.ferriphilum在黄铁矿培养体系(53.8%)和硫酸亚铁和硫粉为能源的培养体系中(45.9%)中丰度最高;在以黄铜矿为能源物质的培养体系中,S.thermotolerans的比例大幅上升(70.1%)。关键词:中度嗜热细菌;生物多样性;生物浸矿;喜温硫杆菌(Acidithiobacillus caldus);耐温氧化硫化杆菌(Sulfobacillus thermotolerans);嗜铁钩端螺旋菌(Leptospirillumferriphilum)     

嗜酸性硫杆菌群体感应系统研究进展

嗜酸性硫杆菌(Acidithiobacillus spp.)是一类重要的极端环境微生物与工业微生物。该类细菌通过氧化硫或亚铁获得电子以固定二氧化碳进行自养生长,是驱动矿山环境酸化和重金属溶出的关键菌群,也是生物冶金等微生物浸出技术中的核心菌群。群体感应(quorum sensing, QS)系统是细菌种内及种间信息交流的重要方式,广泛分布于嗜酸性硫杆菌等化能自养微生物中,比如类似于LuxI/R的AfeI/R系统。系统介绍近年来嗜酸性硫杆菌菌体感应系统研究成果,尤其是在AfeI/R种群分布、生物学功能、调节机制及其应用研究中的新发现与新理论。讨论今后嗜酸性硫杆菌群体感应系统研究的主要方向及需要解决的关键科学问题,以促进极端微生物群体感应系统理论研究的开展与产业应用技术的开发。     

嗜酸硫杆菌在工农业中的应用

嗜酸硫杆菌(属)(Acidithiobacillus spp.)能够氧化亚铁、硫或还原性无机硫化合物(reduced inorganic sulfur compounds,RISCs)获得能量,固定二氧化碳,是一类典型的嗜酸性化能自养微生物。嗜酸硫杆菌广泛分布于酸性矿水、热泉等酸性环境中,是地球生态系统硫和铁元素循环的主要推动者。嗜酸硫杆菌独特的生理代谢特征和极端环境适应性,使其广泛应用于生物浸出领域。本文综述了嗜酸硫杆菌的生理代谢特征和极端环境下的适应机制,阐述了嗜酸硫杆菌在工农业中的应用,讨论了面向国家重大需求,嗜酸硫杆菌在今后的主要研究方向和需要解决的关键科学问题,为嗜酸硫杆菌在生理代谢、环境适应和工农业应用的研究提供重要的线索和启示。     

一株新分离巴氏杆菌的初步鉴定

对清洁级昆明小鼠常规监测中分离的一株可疑嗜肺巴氏杆菌进行了有关遗传学和生物表型特征的检定。该菌形态学及一般培养特性与嗜肺巴氏杆菌相似 ,与嗜肺巴氏杆菌参考血清发生凝集反应 ,生化试验不符合嗜肺巴氏杆菌典型特征 ,染色体DNA的G -Cmol%测定不能排除巴斯德氏菌属细菌的可能 ,进一步生化检定不符合国外报道的嗜肺巴氏杆菌Jawetz和Hey 1 .2个生物型 ,但与国外报道的Pasteurella .sp组细菌相似。     

氧化亚铁硫杆菌的分离及其培养条件优化

目的:获得可应用于烟气脱硫的菌株,并对其培养条件进行优化.方法:从化工厂取土样分离氧化亚铁硫杆菌,分析分离菌株的形态学特征、培养特征及16S rDNA序列,确定菌株的分类地位.通过单因子实验,对培养基中主要成分硫酸亚铁和硫酸铵的浓度进行优化.利用SAS软件中的Box-Behnken法设计实验,通过响应面分析对初始pH、温度、接种量、装液量4个因素进行优化.结果:获得菌株N16,经鉴定为嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans).确定FeSO_4·7H_2O和(NH_4)_2SO_4的最适添加量分别为60/L和1g/L.确定菌株最适培养条件为:初始pH 1.8,温度28℃,转速150r/min,接种量15%,装液量30mL.在最适培养墓及培养条件下,菌株N16的亚铁氧化率可达99.78%.结论:分离得到的菌株适合于微生物法烟气脱硫的应用.     

Cooperation between two Thiobacillus strains for heavy-metal removal from municipal sludge.

A mixed culture of two fast-growing bacterial strains for heavy-metal solubilization of municipal sewage sludge has been developed. Strain VA-7 decreases the initial sludge pH (7-8.5) to a value between 4.0 and 4.5. Then, strain VA-4 begins growing and further reduces the pH to values below 2.0. The rapid decrease of sludge pH by a mixed culture through sulfur oxidation into sulfuric acid solubilizes the toxic metals (Cd 83-96%, Cr 16-54%, Cu 85-87%, Mn 91-94%, Ni 78-79%, Pb 28-46%, Zn 82-96%) to levels recommended for intensive use of residual sludge in agriculture. A study of the physiological and metabolic characteristics of these strains revealed that isolate VA-7 is a strain of Thiobacillus thioparus (ATCC 55127), while isolate VA-4 corresponds to a Thiobacillus thiooxidans (ATCC 55128). These bacterial strains possess distinctive physiological characteristics that allow them to easily grow and solubilize heavy metals in municipal sludge.     

柠檬酸、草酸和乙酸对污泥中镉、铅、铜和锌的去除效果

研究了柠檬酸、草酸和乙酸对污泥中Cd、Pb、Cu和Zn等重金属的去除效果,并分析了污泥中重金属的形态变化和生物有效性.结果表明,随着反应时间和酸浓度的增加,污泥中重金属（Cu除外）的去除率也相应增加.污泥加入柠檬酸溶液反应7 h即可去除污泥中52.0%的Pb和74.2％的Zn, 24 h后可去除76.0%的Pb和92.5%的Zn,草酸和乙酸对重金属的去除率较低.柠檬酸去除的Pb和Zn主要以稳定态存在,并导致污泥中不稳定态重金属的比例上升,其中可交换态重金属浓度有不同程度的增加.虽然有机酸对Cd和Cu的去除率较低,但反应后可交换态Cd和Cu的浓度仍有小幅增加.     

利用乙二胺四乙酸淋洗修复重金属污染的土壤及其动力学

通过室内模拟试验,采用振荡淋洗的方法研究了乙二胺四乙酸(EDTA)浓度、pH、淋洗时间对重金属去除效果的影响.利用一级反应动力学模型对试验数据进行拟合,并测定了EDTA处理前后土壤中重金属形态的变化.结果表明,EDTA溶液在浓度为0.1 mol·L^-1、pH 7、淋洗时间1 d的条件下能达到对污染土壤重金属的最大去除率,去除率分别为Cd 89.14%、Pb 34.78%、Cu 14.96%、Zn 45.14%.模型拟合结果表明,Cd的质量转移系数最大,其次是Zn、Pb和Cu.说明在土壤淋洗过程中,Cd和Zn最先达到质量转移的平衡状态,然后是Pb和Cu.形态分级结果表明,EDTA能有效地去除交换态、碳酸盐结合态和氧化物结合态重金属,而对有机态和残余态部分重金属作用效果不明显.     

Effects of Municipal Sewage Sludge Doses on the Chlorophyll Contents and Heavy Metal Concentration of Sugar Beet (Beta vulgaris var. saccharifera)

The present study was conducted to assess the suitability of sewage sludge amendment (SSA) in soil for Beta vulgaris var. saccharifera (sugar beet) by evaluating the heavy metal accumulation and physiological responses of plants grown at a 10%, 25%, and 50% sewage sludge amendment rate. The sewage sludge amendment was modified by the physicochemical properties of soil, thus increasing the availability of heavy metals in the soil and consequently increasing accumulation in plant parts. Cd, Pb, Ni, and Cu concentrations in roots were significantly higher in plants grown at 25% as compared to 50% SSA; however, Cr and Zn concentration was higher at 50% than 25% SSA. The concentrations of heavy metal showed a trend of Zn > Ni > Cu > Cr > Pb > Cd in roots and Zn > Cu > Ni > Cr > Pb > Cd in leaves. The only instance in which the chlorophyll content did not increase after the sewage sludge treatments was 50%. There were approximately 1.12-fold differences between the control and 50% sewage sludge application for chlorophyll content. The sewage sludge amendment led to a significant increase in Pb, Cr, Cd, Cu, Zn, and Ni concentrations of the soil. The heavy metal accumulation in the soil after the treatments did not exceed the limits for the land application of sewage sludge recommended by the US Environmental Protection Agency (US EPA). The increased concentration of heavy metals in the soil due to the sewage sludge amendment led to increases in heavy metal uptake and the leaf and root concentrations of Ni, Zn, Cd, Cu, Cr, Pb, and Zn in plants as compared to those grown on unamended soil. More accumulation occurred in roots and leaves than in shoots for most of the heavy metals. The concentrations of Cd, Cr, and Pb were more than the permissible limits of national standards in the edible portion of sugar beet grown on different sewage sludge amendment ratios. The study concludes that the sewage sludge amendment in the soil for growing sugar beet may not be a good option due to risk of contamination of Cr, Pb, and Cd.     

Leaching Heavy Metals from Contaminated Soil by Using Thiobacillus ferrooxidans or Thiobacillus thiooxidans

Iron- and sulfur-oxidizing bacteria identified as Thiobacillus ferrooxidans and T. thiooxidans were successfully enriched from various soil samples contaminated with heavy metals and organic compounds. Depending on the growth medium, the soil sample, and the type of contaminant, the indigenous isolates solubilized > 50% of most of the heavy metals present in the solid sample (As, Cd, Co, Cr, Cu, Ni, V, Zn, B, Be). Leaching with T. ferrooxidans strains resulted in total extraction of Cd, Co, Cu, and Ni. With sulfur-oxidizing bacteria > 80% of Cd, Co, Cu, and Zn was mobilized from rainwater sludge. Pb and Ba were not detected in the leachate, given the insolubility of their sulfate compounds. An increase in pulp density up to 20%, indicating 6.6% total organic carbon in the soil and rubble leach experiment (sample 557), did not inhibit the growth of the indigenous T. ferrooxidans strain. In view of these results, bioleaching appears to have some potential for remediation of heavy metal contaminated soils.     

Inter- and intraspecific genomic variability of the 16S-23S intergenic spacer regions (ISR) in representatives of Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans

The complete sequences of 32 intergenic spacer regions (ISR) from Acidithiobacillus strains, including 29 field strains isolated from coal, copper, molybdenum mine wastes or sediment of different geoclimatic regions in China, reference strain ATCC19859 and the type strains of the two species were determined. These data, together with other sequences available in the GenBank database, were used to carry out the first detailed assessment of the inter- and intraspecific genomic variability of the ISR sequences and to infer phylogenetic relationships within the genus. The total length of the 16S-23S rRNA intergenic spacer regions of the Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans strains ranged from 451 to 490 bp, and from 434 to 456 bp, respectively. The degree of intrageneric ISR sequence similarity was higher than the degree of intergeneric similarity, and the overall similarity values of the ISRs varied from 60.49% to 84.71% between representatives of different species of the genus Acidithiobacillus. Sequences from the spacer of the A. thiooxidans and A. ferrooxidans strains ranged from 86.71% to 99.56% and 92.36% to 100% similarity, respectively. All Acidithiobacillus strains were separated into three phylogenetic major clusters and seven phylogenetic groups. ISR may be a potential target for the development of in situ hybridization probe aimed at accurately detecting acidithiobacilli in the various acidic environments.     

Mobility and Distribution of Zinc,Cadmium and Lead in Calcareous Soils Receiving Spiked Sewage Sludge

Leaching column experiments were conducted to determine the degree of mobility and the distribution of zinc (Zn), cadmium (Cd), and lead (Pb) because of an application of spiked sewage sludge in calcareous soils. A total of 20 leaching columns were set up for four calcareous soils. Each column was leached with one of these inflows: sewage sludge (only for two soils), spiked sewage sludge, or artificial well water (control). The columns were irrigated with spiked sewage sludge containing 8.5 mg Zn l^?1, 8.5 mg Cd l^?1, and 170 mg Pb l^?1 and then allowed to equilibrate for 30 days. At the end of leaching experiments, soil samples from each column were divided into 18 layers, each being 1 cm down to 6 cm and 2 cm below that, and analyzed for total and extractable Zn, Cd and Pb. The fractionation of the heavy metals in the top three layers of the surface soil samples was investigated by the sequential extraction method. Spiked sewage sludge had little effect on metal mobility. In all soils, the surface soil layers (0-1 cm) of the columns receiving spiked sewage sludge had significantly higher concentrations of total Zn, Cd and Pb than control soils. Concentration of the heavy metals declined significantly with depth. The mobility of Zn was usually greater than Cd and Pb. The proportion of exchangeable heavy metals in soils receiving spiked sewage sludge was significantly higher than that found in the control columns. Sequential extraction results showed that in native soils the major proportion of Zn and Pb was associated with residual (RES) and organic matter (OM) fractions and major proportion of Cd was associated with carbonate (CARB) fraction, whereas after leaching with spiked sewage sludge, the major proportion of Zn and Pb was associated with Fe-oxcide (FEO), RES, and CARB fractions and major proportion of Cd was associated with CARB, RES and exchangeable (EXCH) fractions. Based on relative percent, Cd in the EXCH fraction was higher than Zn and Pb in soils leached with spiked sewage sludge.     

Accumulation of heavy metals in rainbow trout Salmo gairdneri (Richardson) maintained on a diet containing activated sewage sludge

Rainbow trout were fed for 10 weeks with a nutritionally balanced diet containing 30% by weight of activated sewage sludge. The whole body concentrations of nine heavy metals (Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, Pb), together with four major cations (Ca, Mg, Na, K) were determined at the beginning and end of the experiment and at three intermediate stages. Fish fed on the diet containing sewage sludge had significantly elevated levels of Cr, Fe, Ni, Pb and reduced levels of Na and K compared with controls, though the values obtained for all groups fell within the range reported for uncontaminated fish. The Ni and Zn showed a marked increase towards the end of the experiment, suggesting that they might have continued to rise after 70 days.     

Bio-Acidification and Leaching of Metals,Nitrogen, and Phosphorus from Soil and Sludge Mixtures

Soil and wastewater treatment sludge are commonly brought together in mixtures for a variety of beneficial purposes. The mixtures contain bioacidifying (i.e., sulfur-oxidizing) microorganisms that can easily be activated through providing the appropriate substrate and environmental conditions. In this study, contaminated soil and sludge mixtures were subjected to controlled bio-acidification and the impacts of the process on the partitioning of heavy metals, nitrogen, and phosphorus were examined. Three successive bio-acidification cycles resulted in significant leaching of metals from sludge. The leaching results, expressed as fraction of total mass of metals in the sludge, averaged 67% for Cr, 96% for Ni, 24% for Zn; 16% for Cu; 23% for Cd; and 96% for Pb. Bio-acidification of the sludge also converted 28 to 45% of the organic nitrogen into ammonia and increased the soluble orthophosphates fraction of total phosphorus by approximately 18 to 20%. Bio-acidification also resulted in significant metals leaching from the contaminated soils in the soil/sludge mixtures. Soil/sludge mixtures were prepared using six soil particle sizes (less than 0.075?mm to 2.38?mm) contaminated with 22,500?mg/kg Zn, 14,000?mg/kg Pb, 1500?mg/kg Cr, 9500?mg/kg Cu, 1000?mg/kg Ni, and 1000?mg/kg Cd. The addition of metals to the soil inhibited the sulfur-oxidizing microorganisms, preventing bio-acidification in the mixtures containing 4 to 50?g soil in 130?ml sludge, and considerably slowing bio-acidification in the mixtures containing 1 to 3?g soil. Using a mixture that contained 2-g soil samples, three successive bio-acidification cycles resulted in significant cumulative metals leaching results. The leaching results, expressed as percentage of the mass of metals added to the soil, were in the range of 56 to 98% for Cr, 77 to 95% for Zn, 33 to 66% for Ni, 64 to 82% for Cu, and 10 to 33% for Pb, with the higher results in each range belonging to the larger size soil particles. On the other hand, only Cr was leached in neutralized soil samples. The results confirmed the potential for inhibition of the sulfur-oxidizing microorganisms and bio-acidification in contaminated soil/sludge mixtures, and the significant impacts of bio-acidification on the mobility of metals, nitrogen, and phosphorus. In addition, the results confirmed the potential for using controlled bioacidification for removing heavy metals from contaminated soil using the indigenous sulfur oxidizing microorganisms in sludge.