应用于PCR-DGGE分析的活性污泥微生物总DNA提取方法比较

探讨适用于PCR-DGGE分析研究的活性污泥细菌和真菌的DNA提取方法。采用5种方法提取活性污泥微生物基因组DNA,以DNA纯度、含量、片段大小及DGGE条带多样性作为考察指标评价提取方法的优劣,以确定最佳实验方案。紫外吸收法和琼脂糖凝胶电泳结果显示,试剂盒法提取的DNA含量最低,其余4种方法获得的DNA含量无显著差异,就DNA纯度而言,试剂盒法最优;除高温裂解法对真菌细胞壁裂解效果较差外,其他4种方法均能不同程度地裂解细菌和真菌细胞;DGGE结果表明,高温裂解法获得的细菌条带最多,基于SDS的细胞裂解法得到的真菌条带最多。综合分析,高温裂解法更适合于活性污泥中细菌的PCR-DGGE分析,基于SDS的细胞裂解法则更适合于污泥中真菌的PCR-DGGE分析。     

2种分步浸提方法对土壤中Cr形态提取效果的比较

实验模拟了受铬渣污染的土壤环境,比较在不同活性炭加入量下2种典型分步浸提方法:Tessier 5步法和改进的7步分离方法得到的不同结果,探究适合于铬渣污染对土壤环境的铬形态浸提方法。实验结果表明,2种方法的浸提数据虽有较大差异,但能得到相对一致的变化趋势;七步法形态分类更为细致,99%以上的铬都分布在2~3个结合形态,Tessier五步法的物料衡算结果更为准确(96%~103%),相比七步法形态分类更具包容性,数据更具重现性;随着活性炭的加入,土壤环境中游离铬离子明显减少,转化为有机结合态等更稳定的结合形态,说明活性炭能有效固定土壤环境中游离的铬离子,增加其迁移稳定性。     

Leaching Characteristics of Heavy Metals and As from Two Urban Roadside Soils

The leaching tests, including the Toxicity Characteristic Leaching Procedure (TCLP), EDTA extraction and BCR sequential extractions before and after EDTA treatments, were performed on two specific soils to elucidate heavy metal-associated mineral fractions and general leachability. The TCLP illustrated the low leachability of heavy metals in soils from two sites. EDTA is a strong chelator and therefore had higher extraction efficiency compared to that of TCLP. The lower extraction percentages by EDTA for As and Sb were found compared to the other heavy metals derived from anthropogenic sources. Sequential extractions showed that the importance of acid-extractable, organically-bound and Fe–Mn oxide fractions was identified for anthropogenic heavy metals with the exception of As and Ni while the importance of residual fraction was identified for endogenous metals. Changes in sequential fractions of heavy metals after leaching with EDTA are very complex and it is difficult to generalize on which fraction was more mobile than the others. These combined results are helpful in elucidating the association of heavy metals to soil fractions and the mobility characteristics of heavy metals under certain environmental conditions.     

Environmentally friendly assessment of organic compound bioaccessibility using sub-critical water

The evaluation of microbial availability of contaminants is of high importance for better reflecting the processes governing contaminant fate in soil and for establishing the risk associated with contaminated sites. A sub-critical water extraction technique was assessed for its potential to determine the microbially degradable fraction of [¹⁴C]phenanthrene-associated activity in two dissimilar soils at three different ageing times (14, 28 and 49 days). For the majority of determinations, no significant (p > 0.05) difference between sub-critical water-extracted ¹⁴C-activity at 160 °C and the fraction mineralized by catabolically active Pseudomonas sp. was observed. Collectively, the results suggested that the sub-critical water extraction technique was an appropriate technique for predicting the biodegradable fraction of phenanthrene-associated ¹⁴C-activity in dissimilar soils following increasing soil-contaminant contact time.     

The impact of synthetic pyrethroid and organophosphate sheep dip formulations on microbial activity in soil

Sheep dip formulations containing organophosphates (OPs) or synthetic pyrethroids (SPs) have been widely used in UK, and their spreading onto land has been identified as the most practical disposal method. In this study, the impact of two sheep dip formulations on the microbial activity of a soil was investigated over a 35-d incubation. Microbial utilisation of [1-(14)C] glucose, uptake of (14)C-activity into the microbial biomass and microbial numbers (CFUs g(-1) soil) were investigated. In control soils and soils amended with 0.01% sheep dip, after 7d a larger proportion of added glucose was allocated to microbial biomass rather than respired to CO(2). No clear temporal trends were found in soils amended with 0.1% and 1% sheep dips. Both sheep dip formulations at 0.1% and 1% concentrations resulted in a significant increase in CFUs g(-1) soil and [1-(14)C] glucose mineralisation rates, as well as a decline in microbial uptake of [1-(14)C] glucose, compared to control and 0.01% SP- or OP-amended soils. This study suggests that the growth, activity, physiological status and/or structure of soil microbial community may be affected by sheep dips.     

亚临界丙烷萃取条件对人参提取物中有机氯农药残留脱除的影响

以人参提取物为原料,利用亚临界丙烷萃取技术脱除其中的有机氯农药残留,研究了一定范围内的萃取压力(4～12 Mpa)、萃取温度(50～80 ℃)、萃取时间(60～180 min)和溶剂流量(1 96～3 92 kg/h)对亚临界丙烷萃取脱除人参提取物中农药残留的影响.由正交实验确定了最佳萃取条件: 萃取压力8 Mpa,萃取温度65 ℃,萃取时间120 min,溶剂流量2.94 kg/h.在此条件下,γ-BHC、op-DDT残留的脱除率接近100%,TCNB、pp-DDT脱除率大于60%,人参皂甙损失小于6.53%.     

Distribution and variability of redox zones controlling spatial variability of arsenic in the Mississippi River Valley alluvial aquifer, southeastern Arkansas

Twenty one of 118 irrigation water wells in the shallow (25-30 m thick) Mississippi River Valley alluvial aquifer in the Bayou Bartholomew watershed, southeastern Arkansas had arsenic (As) concentrations (<0.5 to 77 microg/L) exceeding 10 microg/L. Sediment and groundwater samples were collected and analyzed from the sites of the highest, median, and lowest concentrations of As in groundwater in the alluvial aquifers located at Jefferson County, Arkansas. A traditional five-step sequential extraction was performed to differentiate the exchangeable, carbonate, amorphous Fe and Mn oxide, organic, and hot HNO(3)-leachable fraction of As and other compounds in sediments. The Chao reagent (0.25 M hydroxylamine hydrochloride in 0.25 M HCl) removes amorphous Fe and Mn oxides and oxyhydroxides (present as coatings on grains and amorphous minerals) by reductive dissolution and is a measure of reducible Fe and Mn in sediments. The hot HNO(3) extraction removes mostly crystalline metal oxides and all other labile forms of As. Significant total As (20%) is complexed with amorphous Fe and Mn oxides in sediments. Arsenic abundance is not significant in carbonates or organic matter. Significant (40-70 microg/kg) exchangeable As is only present at shallow depth (0-1 m below ground surface). Arsenic is positively correlated to Fe extracted by Chao reagent (r=0.83) and hot HNO(3) (r=0.85). Arsenic extracted by Chao reagent decreases significantly with depth as compared to As extracted by hot HNO(3). Fe (II)/Fe (the ratio of Fe concentration in the extracts of Chao reagent and hot HNO(3)) is positively correlated (r=0.76) to As extracted from Chao reagent. Although Fe (II)/Fe increases with depth, the relative abundance of reducible Fe decreases noticeably with depth. The amount of reducible Fe, as well as As complexed to amorphous Fe and Mn oxides and oxyhydroxides decreases with depth. Possible explanations for the decrease in reducible Fe and its complexed As with depth include historic flushing of As and Fe from hydrous ferric oxides (HFO) by microbially-mediated reductive dissolution and aging of HFO to crystalline phases. Hydrogeochemical data suggests that the groundwater in the area falls in the mildly reducing (suboxic) to relatively highly reducing (anoxic) zone, and points to reductive dissolution of HFO as the dominant As release mechanism. Spatial variability of gypsum solubility and simultaneous SO(4)(2-) reduction with co-precipitation of As and sulfide is an important limiting process controlling the concentration of As in groundwater in the area.     

Multiple batch extraction test to estimate contaminant release parameters using a Bayesian approach

Industrial activities produce vast amounts of weakly contaminated materials which are commonly reused as filling materials on natural ground. There is a strong demand to define guidelines for the application of these materials, to estimate the leaching potential of contaminants from the materials, and to assess the potential hazard for groundwater pollution. We present a multiple batch experiment, where measurements of liquid-phase concentrations at varying liquid/solid ratios are used to estimate the total mass of contaminant that can be extracted from a contaminated material with a mild extractant like water. Furthermore, the experiment yields estimates of the isotherm describing the partitioning of the contaminant between the solid and liquid phases, and a concentration that might be expected under soil hydraulic conditions representative for the field situation. Model parameters are estimated from liquid-phase concentrations within a Bayesian framework by applying the Shuffled Complex Evolution Metropolis Algorithm (SCEM-UA), an efficient Markov Chain Monte Carlo sampler. A sensitivity analysis and inversions of synthetically generated data corrupted with noise show the general suitability of the proposed method. An uncertainty analysis for model parameters and model predictions shows the expected accuracy of the estimates. An application to concentration measurements obtained from a multiple batch extraction test illustrates the applicability of the approach for a real situation.     

Use of the physiologically-based extraction test to assess the oral bioaccessibility of metals in vegetable plants grown in contaminated soil

The oral bioaccessibility of metals in vegetable plants grown on contaminated soil was assessed. This was done using the physiologically-based extraction test (PBET) to simulate the human digestion of plant material. A range of vegetable plants, i.e. carrot, lettuce, radish and spinach, were grown on metal contaminated soil. After reaching maturity the plants were harvested and analysed for their total metal content (i.e. Cr, Cd, Cu, Fe, Mn, Mo, Ni, Pb and Zn) by inductively coupled plasma-mass spectrometry (ICP-MS). The plant samples were then subsequently extracted using an in vitro gastrointestinal approach or PBET to assess the likelihood of oral bioaccessibility if the material was consumed by humans.     

蜈蚣草总黄酮提取工艺的优选

采用水浴法提取总黄酮,以蜈蚣草为原料、总黄酮得率为考察指标,通过正交试验研究乙醇浓度、提取时间、提取温度及料液比对提取蜈蚣草总黄酮得率的影响。结果表明,各因素对总黄酮提取率的影响程度为∶乙醇浓度提取时间提取温度料液比;最佳的提取工艺条件为:乙醇浓度60%,料液比1∶15,提取温度60℃,每次3h,提取3次,在此条件下提取的总黄酮得率为6.15%。