DGGE、T-RFLP 、LH-PCR对两种活性污泥的微生物种群多样性分析的比较

采用基于PCR扩增的分子生态技术DGGE、T-RFLP和LH-PCR,对2种典型污水处理系统中的活性污泥（生活污泥、焦化污泥）进行微生物种群多样性分析;并以此比较3种技术的优劣,提出不同应用条件下研究方法的选择依据。根据实验结果：DGGE得到的条带较多,但误差来源也最多;T-RFLP技术较为灵敏,但需要选择适当的内切酶,严格控制酶切条件,并且文库比对误差较大;而LH-PCR操作简单,结果稳定性较高。虽然目前尚无法判断3种方法的准确性,但LH-PCR在活性污泥的微生物种群多样性分析中已显示出潜在优势。     

Linkage between community diversity of sulfate-reducing microorganisms and methylmercury concentration in paddy soil

Sulfate-reducing microorganisms (SRM) have been thought to play a key role in mercury (Hg) methylation in anoxic environments. The current study examined the linkage between SRM abundance and diversity and contents of methylmercury (MeHg) in paddy soils collected from a historical Hg mining area in China. Soil profile samples were collected from four sites over a distance gradient downstream the Hg mining operation. Results showed that MeHg content in the soil of each site significantly decreased with the extending distance away from Hg mine. Soil MeHg content was correlated positively with abundance of SRM and the contents of organic matter (OM), NH₄ ⁺, SO₄ ^2?, and Hg. The abundances of SRM based on dissimilatory (bi) sulfite reductase (dsrAB) gene at 0–40 cm depths were higher than those at 40–80 cm depth at all sites. The SRM community composition varied in the soils of different sampling sites following terminal restriction fragment length polymorphism (T-RFLP) and phylogenetic analyses, which appeared to be correlated with contents of MeHg, OM, NH₄ ⁺, and SO₄ ^2? through canonical correspondence analysis. The dominant groups of SRM in the soils examined belonged to Deltaproteobacteria and some unknown SRM clusters that could have potential for Hg methylation. These results advance our understanding of the relationship between SRM and methylmercury concentration in paddy soil.     

The effect of heavy metals on the activated sludgeprocess and its microbial community analysis using16S ribosomal DNA

The effect of heavy metal shock loading on biological treatment systems was studied by traditional methods and molecular biological techniques. Two kinds of SBR (sequence batch reactor) operation units, unacclimated and acclimated activated sludge systems, were studied. The addition of special nutrients and powdered activated carbon (PAC) to stimulate heavy metal uptake and recovery were studied. The kinetic constants could be used to describe the effect of the inhibition of substance utilisation. The results showed that heavy metal shock loading had a greater effect on the unacclimated activated sludge system than on the acclimated one. The special nutrients greatly enhanced the uptake of copper, and the PAC improved sludge settling and decreased the turbidity of the effluent. The variation of dominant species and the diversity of the bacterial community were analysed using 16S ribosomal DNA. Compared with the slight change of dominant species during acclimation by copper, there was a great change in the acclimated system shocked by a high concentration of copper. The results confirmed that the acclimation could improve the resistance of microorganisms to heavy metal toxicity.     

煤化工废水生化处理系统活性污泥膨胀控制及菌群迁移

针对煤化工废水生化处理系统存在的活性污泥丝状菌膨胀问题,根据微生物营养学原理,利用活性污泥调理剂对污泥膨胀进行控制和修复,并对修复前后的活性污泥细菌菌群进行高通量测序,研究了菌群的迁移变化途径。研究结果表明,修复后的煤化工废水生化处理系统中活性污泥的SVI值明显下降,维持在85 mL·g-1左右,系统对废水中有机污染物的降解能力有所提升,COD去除率由原来的93%左右提高到96%以上。细菌菌群种类、数量和丰度都有明显改善。     

Effect of methamidophos and urea application on microbial communities in soils as determined by microbial biomass and community level physiological profiles

In this study, we evaluated the effect of the application by two agrochemicals, methamidophos (O,S-dimethyl phosphoroamidothioate) and urea, on microbial diversity in soil, using the combined approaches of soil microbial biomass analysis and community level physiological profiles (CLPPs). The results showed that both a low and a high level of methamidophos application (CS2 and CS3) and urea application (CS4) significantly decreased microbial biomass C (Cmic) by 41-83% compared with the control (CS1). The soil organic C (Corg) values of CS3 and CS4 were significantly higher and lower by 24% and 14%, respectively, than that of CS1. Similarly to Cmic, the values of Cmic/Corg of the three applied soils which decreased were lower by 31-84% than that of CS1. In contrast, the respiration activity of the three applied soils were significantly higher than the control. Agrochemical application also significantly increased the soil total of N and P (Ntol and Ptol) and decreased the Corg/Ntol and Corg/Ptol values. The CLPPs results showed that the AWCD (average well color development) of the three applied soils were significantly higher than that of CS1 during the incubation period. Substrate richness, Shannon and Simpson indices of microbial communities under chemical stresses, increased significantly. In addition, the CFU (colony-forming unit) numbers of methamidophos metabolized bacteria in CS2 and CS3 also increased significantly by 86.1% and 188.9% compared with that of CS1. The combined results suggest that agrochemicals reduce microbial biomass and enhance functional diversities of soil microbial communities; meanwhile, some species of bacteria may be enriched in soils under methamidophos stress.     

Comparison of the purification performance and microbial community functional diversity in flow-directional-switching and unidirectional-flow biotrickling filters

Because of the characteristics of low operating cost and convenient operation, the biotrickling filter is extensively researched and used to treat low concentration waste gas contaminated by volatile organic compounds (VOCs) and other odors. In this paper, two laboratory-scale biotrickling filters were constructed and toluene was selected as the sole carbon source, and the effects of different waste-gas flow configuration patterns on the purification capacity and the microbial community functional diversity of biotrickling filters were evaluated. The results indicated that the flow-directional-switching (FDS) biotrickling filter had better purification performance, and the maximum elimination capacity reached 480 g·m^?3·hr^?1, which was 17.1% higher than conventional unidirectional-flow (UF) biotrickling filter. Comparing the purification capacities of different sections in two biotrickling filters, the maximum toluene elimination capacity of section III in FDS system could reach 542 g·m^?3·hr^?1, which was 2.8 times as great as that in UF system, which resulted from the difference of elimination capacity in two systems. By analyzing the metabolic activity of two systems by community-level physiological profiling (CLPP) with Biolog (Biolog Inc., Hayward, CA) ECO-plate technique, metabolic activity in three sections of FDS system was higher than that of UF system. The metabolic activity was the highest in section III of FDS system and 46.8% higher than that of UF system. Shannon index and McIntosh index of section III in FDS system were 6.2% and 31.5% higher, respectively, than those of UF system.

Implications: The flow-directional-switching (FDS) biotrickling filter had a better purification performance than unidirectional-flow (UF) biotrickling filter at high inlet loadings, because FDS produced a more uniform distribution of biomass and microbial metabolic capacity along the length of the packed bed without diminishing activity and removal capacity in the inlet section.     

光合细菌改善Cd、Pb及呋喃丹污染土壤的微生物群落DNA序列多样性的研究

应用DNA随机扩增多态性(RAPD)分子标记技术研究了光合细菌(PSB)对Cd、Pb及呋喃丹污染土壤的微生物群落DNA序列多样性的影响.结果表明,Cd、Pb及呋喃丹单一污染或3者复合污染土壤的微生物群落DNA序列的丰富度相对对照土样(S0)都有不同程度的增加,受Cd、Pb或呋哺丹污染,可能会引起土壤微生物群落DNA序列...     

Insecticide fingerprinting tecinique for detection and location of organochlorine insecticide rssiduss in foods

Abstract

Insecticide fingerprinting technique enables the detection and location of DDT and HCII residues in vegetables through the development of green and Prussian blue colors respectively. Cut vegetables are pressed against o‐tolidine impregnated paper and exposed to sunlight where colored spots appear instantly. The studies on 18 vegetable varieties revealed the pesticide residues and their distribution in different tissues. This direct method is sensitive (0.3 μg for HCII & 0.5 μg for DDT) and has special applications in quality control laboratorios and food industry.     

Organochlorine pesticides in the surrounding soils of POPs destruction facility: source fingerprinting,human health,and ecological risks assessment

Environmental Science and Pollution Research - The elimination of persistent organic pollutants (POPs) obsolete pesticides stockpiles, particularly the organochlorine pesticides (OCPs), is one of...     

Temporal trends and sources of PCDD/Fs, pentachlorophenol and chlornitrofen in paddy field soils along the Yoneshiro River basin, Japan

In order to understand the long-term behaviors of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), pentachlorophenol (PCP), and 2,4,6-trichlorophenyl-4′-nitrophenyl ether (chlornitrofen, CNP) in paddy soil, we measured their concentrations in paddy soil samples collected in 1982 and 1984 (1980s) and in 2000 and 2002 (2000s) from the Yoneshiro River basin, Japan. The concentrations of PCP and CNP decreased from the 1980s to the 2000s, whereas the concentrations of PCDD/Fs and their toxic equivalency (WHO2006-TEQ) remained. The major sources of PCDD/Fs in the paddy soil samples were attributed to impurities in PCP and CNP as a result of comparisons of homologue and congener profiles and principal component analysis. Based on the results of comparison of total input and remaining amount, it is estimated that more than 99% of PCP and CNP applied to the paddy fields had disappeared, whereas most of the applied PCDD/Fs and TEQ remained.     

Shift of microbial diversity and function in high-efficiency performance biotrickling filter for gaseous xylene treatment

ABSTRACT

Xylene is the main component of many volatile industrial pollution sources, and the use of biotechnology to remove volatile organic compounds (VOCs) has become a growing trend. In this study, a biotrickling filter for gaseous xylene treatment was developed using activated sludge as raw material to study the biodegradation process of xylene. Reaction conditions were optimized, and long-term operation was performed. The optimal pH was 7.0, gas-liquid ratio was 15:1 (v/v), and temperature was 25 °C. High-throughput sequencing technique was carried out to analyze microbial communities in the top, middle, and bottom layers of the reactor. Characteristics of microbial diversity were elucidated, and microbial functions were predicted. The result showed that the removal efficiency (RE) was stable at 86%–91%, the maximum elimination capacity (EC) was 303.61 g·m^?3·hr^?1, residence time was 33.75 sec, and the initial inlet xylene concentration was 3000 mg·m^?3, which was the highest known degradation concentration reported. Kinetic analysis of the xylene degradation indicated that it was a very high-efficiency-activity bioprocess. The r_max was 1059.8 g·m^?3·hr^?1, and K_s value was 4.78 g·m^?3 in stationary phase. In addition, microbial community structures in the bottom and top layers were significantly different: Pseudomonas was the dominant genus in the bottom layer, whereas Sphingobium was dominant in the top layer. The results showed that intermediate metabolites of xylene could affect the distribution of community structure. Pseudomonas sp. can adapt to high concentration xylene–contaminated environments.

Implications: We combined domesticated active sludge and reinforced microbial agent on biotrickling filter. This system performed continuously under a reduced residence time at 33.75 sec and high elimination capacity at 303.61 g·m^?3·hr^?1 in the biotrickling reactor for about 260 days. In this case, predomestication combined with reinforcing of microorganisms was very important to obtaining high-efficiency results. Analysis of microbial diversity and functional prediction indicated a gradient distribution along with the concentration of xylene. This implied a rational design of microbial reagent and optimizing the inoculation of different sites of reactor could reduce the preparation period of the technology.     

Soil microbial parameters and luminescent bacteria assays as indicators for in situ bioremediation of TNT-contaminated soils

In situ bioremediation is increasingly being discussed as a useful strategy for cleaning up contaminated soils. Compared to established ex situ procedures, meaningful and reliable approaches for monitoring the remediation processes and their efficiency are of special importance. The subject of this study was the significance of two bioassays for monitoring purposes. The work was performed within the scope of a research project on the in situ bioremediation of topsoil contaminated with 2,4,6-trinitrotoluene (TNT). To evaluate changes within different experimental fields during a 17-month remediation period, the results of soil microbial assays and luminescent bacteria assays were compared with chemical monitoring data. The luminescent bacteria assays showed a significant reduction of the water-soluble soil toxicants in the treated fields. This bioassay proved to be a sensitive screening indicator of toxicity and may effectively aid the ecotoxicological interpretation of chemical monitoring data. Microbial biomass (C(mic)), the metabolic quotient (qCO2), and the ratio of microbial to organic carbon (C(mic)/C(org)) showed a highly significant correlation with total concentrations of TNT in the soil. But, in contrast to luminescent bacteria assays, this approach did not reveal any recovery of the soil at the end of the remediation period. There is clear evidence for persistent adverse effects of chronic TNT contamination on the site-specific microbial community and the local carbon cycle in the soil. The study clearly exhibits the differences between, as well as the complementary value of both bioassay approaches for monitoring short-term and long-term effects of soil contamination and the efficiency of remediation.     

Microbial diversity,community composition and metabolic potential in hydrocarbon contaminated oily sludge: prospects for in situ bioremediation

Microbial community composition and metabolic potential have been explored in petroleum-hydrocarbon-contaminated sludge of an oil storage facility. Culture-independent clone library-based 16S rRNA gene analyses revealed that the bacterial community within the sludge was dominated by the members of β-Proteobacteria (35 %), followed by Firmicutes (13 %), δ-Proteobacteria (11 %), Bacteroidetes (10 %), Acidobacteria (6 %), α-Proteobacteria (3 %), Lentisphaerae (2 %), Spirochaetes (2 %), and unclassified bacteria (5 %), whereas the archaeal community was composed of Thermoprotei (54 %), Methanocellales (33 %), Methanosarcinales/Methanosaeta (8 %) and Methanoculleus (1 %) members. Methyl coenzyme M reductase A (mcrA) gene (a functional biomarker) analyses also revealed predominance of hydrogenotrophic, methanogenic Archaea (Methanocellales, Methanobacteriales and Methanoculleus members) over acetoclastic methanogens (Methanosarcinales members). In order to explore the cultivable bacterial population, a total of 28 resident strains were identified and characterized in terms of their physiological and metabolic capabilities. Most of these could be taxonomically affiliated to the members of the genera Bacillus, Paenibacillus, Micrococcus, Brachybacterium, Aerococcus, and Zimmermannella, while two strains were identified as Pseudomonas and Pseudoxanthomonas. Metabolic profiling exhibited that majority of these isolates were capable of growing in presence of a variety of petroleum hydrocarbons as sole source of carbon, tolerating different heavy metals at higher concentrations (≥1 mM) and producing biosurfactant during growth. Many strains could grow under a wide range of pH, temperature, or salinity as well as under anaerobic conditions in the presence of different electron acceptors and donors in the growth medium. Correlation between the isolates and their metabolic properties was estimated by the unweighted pair group method with arithmetic mean (UPGMA) analysis. Overall observation indicated the presence of diverse groups of microorganisms including hydrocarbonoclastic, nitrate reducing, sulphate reducing, fermentative, syntrophic, methanogenic and methane-oxidizing bacteria and Archaea within the sludge community, which can be exploited for in situ bioremediation of the oily sludge.     

Influence of lead acetate on soil microbial biomass and community structure in two different soils with the growth of Chinese cabbage (Brassica chinensis)

A greenhouse pot experiment was conducted to evaluate the impact of different concentrations of lead acetate on soil microbial biomass and community structure during growth of Chinese cabbage (Brassica chinensis) in two different soils. The field soils were used for a small pot, short-term 60-day growth chamber study. The soils were amended with different Pb concentrations, ranging from 0 to 900mgkg(-1) soil. The experimental design was a 2 soilx2 vegetation/non-vegetationx6 treatments (Pb)x3 replicate factorial experiment. At 60 days the study was terminated and soils were analyzed for microbial parameters, namely, microbial biomass, basal respiration and PLFAs. The results indicated that the application of Pb at lower concentrations (100 and 300mgkg(-1)) as lead acetate resulted in a slight increase in soil microbial biomass, whereas Pb concentrations >500mgkg(-1) caused an immediate gradual significant decline in biomass. However, the degree of impact on soil microbial biomass and basal respiration by Pb was related to management (plant vegetation) or the contents of clay and organic matter in soils. The profiles of 21 phospholipid fatty acids (PLFAs) were used to assess whether observed changes in functional microbial parameters were accompanied by changes in the composition of the microbial communities after Pb application at 0, 300 and 900mg Pbkg(-1) soil. The results of principal component analyses (PCA) indicated that there were significant increases in fungi biomarkers of 18:3omega6c, 18:1omega9c and a decrease in cy17:0, which is an indicator of gram-negative bacteria for the high levels of Pb treatments In a word, soil microbial biomass and community structure, therefore, may be sensitive indicators reflecting environmental stress in soil-Pb-plant system. However, further studies will be needed to better understand how these changes in microbial community structure might actually impact soil microbial community function.     

Long-term effects of aided phytostabilisation of trace elements on microbial biomass and activity, enzyme activities, and composition of microbial community in the Jales contaminated mine spoils

We studied the effectiveness of remediation on microbial endpoints, namely microbial biomass and activity, microbial and plant species richness, of an As-contaminated mine spoil, amended with compost (C) alone and in combination with beringite (B) or zerovalent iron grit (Z), to increase organic matter content and reduce trace elements mobility, and to allow Holcus lanatus and Pinus pinaster growth. Untreated spoil showed the lowest microbial biomass and activity and hydrolase activities, and H. lanatus as sole plant species, whereas the presented aided phytostabilisation option, especially CBZ treatment, significantly increased microbial biomass and activity and allowed colonisation by several plant species, comparable to those of an uncontaminated sandy soil. Microbial species richness was only increased in spoils amended with C alone. No clear correlation occurred between trace element mobility and microbial parameters and plant species richness. Our results indicate that the choice of indicators of soil remediation practices is a bottleneck.     

The application of positive matrix factorization in the analysis, characterisation and detection of contaminated soils

Multivariate factor analytical techniques are widely used for the approximation, in terms of a linear combination of factors, of multivariate experimental data. The chemical composition of soil samples are multivariate in nature and provide datasets suitable for the application of these statistical techniques. Recent developments of multivariate factor analytical techniques have led to the approach of Positive Matrix Factorization (PMF), a weighted least squares fit of a data matrix in which the weights are determined depending on the error estimates of each individual data value. This approach relies on more physically significant assumptions than methods like Principal Components Analysis which is frequently used in the analysis of soil datasets. In this paper we apply PMF to characterise the pollutant source in a set of geographically referenced soil samples taken within a 200 m radius of a site characterised by a high concentration of heavy metals. Each sample has been analysed for major and minor elements (using wavelength-dispersive X-ray fluorescence spectrometry), carbon, hydrogen and nitrogen (using a CHN elemental analyzer) and mercury (using cold-vapour atomic absorption spectrometry). Analysis of the soils using PMF resulted in a successful partitioning of variances into sources related to background soil geochemistry, organic influences and those associated with the contamination. Combining these results with a geostatistical approach successfully demarcated the main source of the combined organic and heavy metal contamination.     

Inhibition of residual <Emphasis Type="Italic">n</Emphasis>-hexane in anaerobic digestion of lipid-extracted microalgal wastes and microbial community shift

Converting lipid-extracted microalgal wastes to methane (CH₄) via anaerobic digestion (AD) has the potential to make microalgae-based biodiesel platform more sustainable. However, it is apparent that remaining n-hexane (C₆H₁₄) from lipid extraction could inhibit metabolic pathway of methanogens. To test an inhibitory influence of residual n-hexane, this study conducted a series of batch AD by mixing lipid-extracted Chlorella vulgaris with a wide range of n-hexane concentration (～10 g chemical oxygen demand (COD)/L). Experimental results show that the inhibition of n-hexane on CH₄ yield was negligible up to 2 g COD/L and inhibition to methanogenesis became significant when it was higher than 4 g COD/L based on quantitative mass balance. Inhibition threshold was about 4 g COD/L of n-hexane. Analytical result of microbial community profile revealed that dominance of alkane-degrading sulfate-reducing bacteria (SRB) and syntrophic bacteria increased, while that of methanogens sharply dropped as n-hexane concentration increased. These findings offer a useful guideline of threshold n-hexane concentration and microbial community shift for the AD of lipid-extracted microalgal wastes.     

Impacts of groundwater level fluctuation on soil microbial community,alkane degradation efficiency and alkane-degrading gene diversity in the critical zone: Evidence from an accelerated water table fluctuation simulation

Environmental Science and Pollution Research - Petroleum hydrocarbons are hazardous to ecosystems and human health, commonly containing n-alkanes and polycyclic aromatic hydrocarbons. Previous...     

生物可降解螯合剂[S,S]-乙二胺二琥珀酸和乳酸乙酯提取土壤中重金属的研究

研究了生物可降解的有机螯合剂[S,S]-乙二胺二琥珀酸([S,S]-EDDS,简写为EDDS)和乳酸乙酯(EL)对重金属污染土壤中重金属的提取效率,并用X射线衍射(XRD)和红外光谱(FT-IR)分析了螯合剂提取对土壤理化性质的影响.结果表明:EDDS对土壤中的Cd、Cu、Pb和Zn有较高的提取率,效果比EL好;EDDS和EL混合提取的重金属提取率增加,但相比单独使用EDDS没有超过10%.FT-IR分析表明,提取前后FT-IR谱图中特征吸收频率没有显著改变,但强度有所改变.XRD分析表明,提取前后土壤晶格结构发生明显的变化,螯合剂提取改变了土壤微观结构.