Corynebacterium glutamicum as an indicator for environmental cobalt and silver stress–A proteome analysis

Cobalt and silver are toxic for cells, but mechanisms of this toxicity are largely unknown. Analysis of Corynebacterium glutamicum proteome from cells grown in control and cobalt or silver enriched media was performed by two dimensional gel electrophoresis (2DE) followed by mass spectrometry. Our results indicate that the cell adapted to cobalt stress by inducing five defense mechanisms: Scavenging of free radicals, promotion of the generation of energy, reparation of DNA, reparation and biogenesis of Fe-S cluster proteins and supporting and reparation of cell wall. In response to the detoxification of Ag⁺ many proteins were up-regulated, which involved reparation of damaged DNA, minimizing the toxic effect of reactive oxygen species (ROS) and energy generation. Overexpression of proteins involved in cell wall biosynthesis (1,4-alpha-glucan branching enzyme and nucleoside-diphosphate-sugar epimerase) upon cobalt stress and induction of proteins involved in energy metabolism (2-methylcitrate dehydratase and 1, 2-methylcitrate synthase) upon silver demonstrate the potential of these enzymes as biomarkers of sub-lethal Ag⁺ and Co toxicity.     

高效氨氧化菌群富集、驯化及其动态变化规律分析

利用选择性培养基对氨氧化菌群进行了连续驯化,得到了氨氮去除效率稳定的氨氧化菌群。采用平板菌落计数法结合聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术考察了氨氧化菌群在连续传代过程中数量及群落结构的动态变化,并考察了pH值、碳源(HCO3-)浓度和氨氮浓度等因素对氨氧化菌群去除氨氮效率的影响。结果表明,通过连续驯化,氨氧化菌的比例由最初的1.8%提高到了31.3%。在碳源浓度为1.5 mg/L,NH4+-N初始浓度为200 mg/L,pH值为8的条件下,菌群对氨氮的去除率达99%以上。     

ELECTROPHORETIC WATER CLARIFICATION TECHNIQUES1

Only very recently has a limited amount of attention been focused directly on electrophoresis as a method for improving water quality. This approach has been proposed primarily as a method for removal of suspended colloidal material, although solutes can also be removed by this method. Very simply electrophoresis is defined as the movement of charged suspended particles in a dc electric field. Several electrophoretic clarification systems and techniques have been developed and evaluated primarily for removing colloidal clay from suspension. The methods should apply equally well to any negatively charged particulate matter. Design for optimal clay removal efficiency and operating cost efficiency has been based upon previous theoretical results and upon modification of the combination filter-el ectrophores is model of Bier. These systems and techniques along with the theoretical developments leading to their design are discussed. Results and conclusions are given for tests that have been concluded concerning the feasibility or practicality of these electrophoretic clarification systems for commercial use. These are viewed in light of various limiting factors such as electrical conductivity of the medium, quality of water desired, colloid concentrations and electrophoretic mobility of the suspended materials.     

Toxicological characterization of the landfill leachate prior/after chemical and electrochemical treatment: A study on human and plant cells

In this research, toxicological safety of two newly developed methods for the treatment of landfill leachate from the Piškornica (Croatia) sanitary landfill was investigated. Chemical treatment procedure combined chemical precipitation with CaO followed by coagulation with ferric chloride and final adsorption by clinoptilolite. Electrochemical treatment approach included pretreatment with ozone followed by electrooxidation/electrocoagulation and final polishing by microwave irradiation. Cell viability of untreated/treated landfill leachate was examined using fluorescence microscopy. Cytotoxic effect of the original leachate was obtained for both exposure periods (4 and 24 h) while treated samples showed no cytotoxic effect even after prolonged exposure time. The potential DNA damage of the untreated/treated landfill leachate was evaluated by the comet assay and cytokinesis-block micronucleus (CBMN) assay using either human or plant cells. The original leachate exhibited significantly higher comet assay parameters compared to negative control after 24 h exposure. On the contrary, there was no significant difference between negative control and chemically/electrochemically treated leachate for any of the parameters tested. There was also no significant increase in either CBMN assay parameter compared to the negative control following the exposure of the lymphocytes to the chemically or electrochemically treated landfill leachate for both exposure periods while the original sample showed significantly higher number of micronuclei, nucleoplasmic bridges and nuclear buds for both exposure times. Results suggest that both methods are suitable for the treatment of such complex waste effluent due to high removal efficiency of all measured parameters and toxicological safety of the treated effluent.     

Investigation of physico-chemical properties and microbial community during poultry manure co-composting process

Co-composting of poultry manure and rubber wood sawdust was performed with the ratio of 2:1(V/V) for a period of 60 days. An investigation was carried out to study the extracellular enzymatic activities and structural degradation utilizing Fourier transform infrared spectroscopy(FT-IR), thermogravimetry and differential thermal analysis(TG/DTA)and scanning electron microscopy(SEM). The microbial succession was also determined by using denaturing gel gradient electrophoresis(DGGE). The compost was able to reach its highest temperature of 71°C at day 3 and stabilized between 30 and 40°C for 8 weeks.CMCase, FPase and β-glucosidase acted synergistically in order to degrade the cellulosic substrate. The xylanase activities increased gradually during the composting and reached the peak value of 11.637 U/g on day 35, followed by a sharp decline. Both Li P and Mn P activities reached their peak values on day 35 with 0.431 and 0.132 U/g respectively. The FT-IR spectra revealed an increase in aromaticity and a decrease in aliphatic compounds such as carbohydrates as decomposition proceeded. TGA/DTG data exhibited significant changes in weight loss in compost samples, indicating degradation of organic matter. SEM micrographs showed higher amounts of parenchyma exposed on the surface of rubber wood sawdust at day 60, showing significant degradation. DGGE and 16 S r DNA analyses showed that Burkholderia sp., Pandoraea sp., and Pseudomonas sp. were present throughout the composting process. Ornithinibacillus sp. and Castellaniella ginsengisoli were only found in the initial stage of the composting, while different strains of Burkholderia sp. also occurred in the later stage of composting.     

Variations of Bacterial Community Structure in Flooded Paddy Soil Contaminated with Herbicide Quinclorac

The denaturing gradient gel electrophoresis (DGGE) method was applied to determine the relative genetic complexity of microbial communities in flooded paddy soil treated with herbicide quinclorac (3,7-dichloro-8-quinoline-carboylic acid). The results obtained showed a significant effect of quinclorac on the development of bacterial populations in soils contaminated with different concentrations of the herbicide at the early time after application. In general, however, the number of populations of the same soil sample treated with the same concentration of the quinclorac differed obviously with increasing incubation time within the early 8 weeks. The scale of differences in banding patterns-showed that the microbial community structures of the quinclorac-treated and non-quinclorac-treated soils were not significantly different after 21 weeks of incubation. Quantification, as demonstrated in this paper, was studied by establishing dose-response relationships. Significant pattern variations were quantified. Prominent DGGE bands were excised, cloned and sequenced to gain insight into the identities of predominant bacterial populations. The majority of DGGE band sequences were related to bacterial genera Clostridium, Sphingobacterium, Xanthomonas and Rhodococcus.     

Bacterial diversity in paclobutrazol applied agricultural soils

The aim of this study was to investigate the bacterial communities on paclobutrazol [(2RS, 3RS)-1-(4-Chlorophenyl)-4, 4-dimethyl-2-(1H-1,2,4-triazol-1-yl) pentan-3-ol]–applied agricultural soils by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rDNA gene fragments. Three different agricultural soil samples were collected from paclobutrazol applied mango and waxapple orchards, peanut fields and untreated rice fields as a control for DGGE analysis. The DGGE pattern of PCR- generated 16S rDNA gene fragments indicated that the bacterial populations from four paclobutrazol–applied soils of peanut fields were closely related to each other and two paclobutrazol–applied soils of mango and waxapple orchards harbored closely related bacterial communities. But, paclobutrazol–free agricultural soils comprised relatively a different bacterial group. However, the bacterial populations of mango and waxapple orchard are completely different from the bacterial communities of peanut field. Further purification and sequence analysis of 40 DGGE bands followed by phylogenetic tree assay showed similar results that soil bacteria from paclobutrazol applied mango and waxapple orchard are phylogenetically related. Based on the phylogenetic analysis, the clone M-4 was clad 100 % (bootstrap value) with Mycobacterium sp. The Mycobacterium sp. has been proved to degrade the phenolic compounds such as phenol, 4-chlorphenol, 2,4-dichlorophenol and paclobutrazol molecule containing chlorobenzene ring.     

Reduction of Cr(VI) by peat and coal humic substances

The reduction of Cr(VI) by humic substances from leonardite and peat was investigated by capillary zone electrophoresis at various pHs. Both humic materials reduced Cr(VI) at pH 5.4, but not at basic pH. The capacity of leonardite humic substances to reduce Cr(VI) was lower than that of peat humic substances. Fe(III) accelerated the reduction of Cr(VI) by peat humic substances, but not by leonardite humic substances. Cr(VI) reduction mechanisms are proposed. The coal humic substances seem more suitable for remediation of Cr(VI)-contaminated sites.     

人工接种堆肥和自然堆肥微生物区系与分子多态性的变化

采用传统培养方法和PCR-DGGE技术研究了人工接种堆肥和自然堆肥微生物群落的演变过程。结果表明:(1)传统培养方法显示,两种堆肥堆制过程中微生物数量均呈“升高—降低—升高—降低”变化趋势,整个堆肥过程中细菌数量占优势。(2)DGGE图谱显示,两种堆肥不同时期存在不同的细菌种群,其条带数量亦呈“升高—降低—升高—降低”变化趋势。堆肥升温期条带丰富但优势条带不明显;堆肥高温期条带数量减少但出现优势条带,表明高温阶段以嗜热菌或耐高温菌为主;堆肥降温期条带数量再次增多;堆肥腐熟期条带数量少且无优势条带,表明腐熟阶段微生物种群数量少且代谢强度趋于平缓。(3)DGGE图谱表明,人工接种菌株成为堆肥高温期优势菌株。人工接种增加了堆肥中微生物总量,提高了堆肥微生物种群多样性,并且促进了堆肥菌群演替,从而缩短堆肥腐熟时间。     

TAR柱前络合、毛细管电泳分析痕量金属离子

1　IntroductionTransitionmetalswereamongthemostconcernelementsinenvironmentalanalysisduetotheirpotentialrolestoplantandanimals[1 ].Theserolesoftransitionmetalsinenvironmentdependedontheirconcentrationinthesoil,waterandlivingthings.Currently ,themostutiliz…