Biodegradation of chloroacetamide herbicides by Paracoccus sp. FLY-8 in vitro

A butachlor-degrading strain, designated FLY-8, was isolated from rice field soil and was identified as Paracoccus sp. Strain FLY-8 could degrade and utilize six chloroacetamide herbicides as carbon sources for growth, and the degradation rates followed the order alachlor > acetochlor > propisochlor > butachlor > pretilachlor > metolachlor. The influence of molecular structure of the chloroacetamide herbicides on the microbial degradation rate was first analyzed; the results indicated that the substitutions of alkoxymethyl side chain with alkoxyethyl side chain greatly reduced the degradation efficiencies; the length of amide nitrogen's alkoxymethyl significantly affected the biodegradability of these herbicides: the longer the alkyl was, the slower the degradation efficiencies occurred. The phenyl alkyl substituents have no obvious influence on the degradation efficiency. The pathway of butachlor complete mineralization was elucidated on the basis of the results of metabolite identification and enzyme assays. Butachlor was degraded to alachlor by partial C-dealkylation and then converted to 2-chloro-N-(2,6-dimethylphenyl)acetamide by N-dealkylation, which subsequently transformed to 2,6-diethylaniline, which was further degraded via the metabolites aniline and catechol, and catechol was oxidized through an ortho-cleavage pathway. This study highlights an important potential use of strain FLY-8 for the in situ bioremediation of chloroacetamide herbicides and their metabolite-contaminated environment.     

丁草胺降解菌的分离鉴定及降解特性的研究

从处理农药生产废水的膜生物反应器中分离到一株能以丁草胺为惟一碳源和能源生长的细菌BD-1,经鉴定为施氏假单孢菌（Pseudomonas stutzeri）。在纯培养的条件下测定了BD-1对丁草胺的降解性能。结果表明,在接种量为菌浓度OD415萨0．2,pH7．0、30℃条件下,BD-1对丁草胺的降解符合一级动力学特征,1．0、10．0和100．0mg·L^-1的丁草胺的降解半衰期分别为0．11、0．60和0．96d。BD-1在不同pH及温度下对丁草胺的降解作用为pH7．0〉pH6．0〉pH8．0,30℃〉20℃〉40℃。GC／MS初步分析结果表明,丁草胺的主要微生物降解产物为2-氯-2’,6’-二乙基乙酰苯胺和2,6-二乙基苯胺。     

Enzymatic synthesis of 4-amino-3,5-diethylphenyl sulfate, a rodent metabolite of alachlor.

Rat liver tissue homogenates were utilized for in vitro enzymatic conversion of 2,6-diethylaniline (DEA) to the important alachlor metabolite 4-amino-3,5-diethylphenyl sulfate (ADEPS), which was also generated as a radiolabeled standard for use in metabolism studies. ADEPS formation in rodents is associated with the production of other reactive metabolites implicated in alachlor rodent carcinogenesis, making dependable access to an ADEPS standard highly desirable. (14)C-DEA was oxidized by rat liver microsomes to (14)C-4-amino-3,5-diethylphenol, which was further converted to ADEPS via addition of the phosphosulfate transferase cofactor adenosine-3'-phosphate-5'-phosphosulfate. Microprobe NMR was used in conjunction with high-resolution mass spectrometry to fully characterize the resulting (14)C-ADEPS and confirm its structure. Because microgram quantities sufficed for full characterization, the enzymatic transformation provides a viable alternative to radiosynthesis of (14)C-ADEPS.     

Studies on lignin removal by soil perfusion technique

Studies were carried out using a soil perfusion apparatus for the removal of lignin from waste water. It was observed that when a medium (pH 7.0) with lignin having about 1200 color units was perfused through a soil column, perfusate contained only 250 to 300 color units after three days of perfusion and pH was reduced to less than 5.0 with 70 to 80% reduction in Total Organic Carbon (TOC). With a sterile soil column the color was not removed and there was no change in TOC content. When the medium was buffered at pH 7.0, there was no reduction in color units. When the pH of the perfusate reduced to less than 5.0, it was also observed that the nitrate content had increased considerably. These results indicated that due to microbial activity lignin molecules might have been biotransformed and adsorbed on the soil column at lower pH. The major group of microorganisms were also isolated from the soil and their significance is discussed in this paper.     

Degradation of alachlor in natural and sludge-amended soils, studied by gas and liquid chromatography coupled to mass spectrometry (GC-MS and HPLC-MS)

Alachlor [2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl)acetamide] is an herbicide used worldwide. The relative rates of disappearance of alachlor, the formation kinetics of alachlor ethane sulfonic acid (ESA), and the formation of other degradation products in two different soils (a soil with natural organic matter and a sludge-amended soil) has been studied. For such a purpose, soil samples were spiked with alachlor at 2.5 mg kg(-1), concentration generally applied in agricultural soils, and were submitted to sunlight, simulating natural field conditions. Extracts were analyzed by GC-MS and HPLC-MS in scan mode. A good correlation was observed between both techniques, and HPLC-MS allowed the determination of two eluting peaks corresponding to the two stereoisomeric forms of alachlor ESA. Degradation of alachlor in the two soils followed first-order kinetics. Half-life in the natural soil was 4.2 +/- 0.1 days, and half-life in the sludge-amended soil was 5.8 +/- 0.8 days. The higher half-life observed in the sludge-amended soil was attributed to the higher sorption of alachlor to this soil compared to the natural soil. The degradation of alachlor in both soils gave rise to the production of alachlor ESA. Its concentration increased during the incubation period, and after 27 days, its concentration was about 0.59 mg kg(-1) in the natural soil and 0.37 mg kg(-1) in the sludge-amended soil. The other two alachlor transformation products were identified using GC-MS, and the abundance of these degradation products increased while alachlor was degraded.     

Dimetridazole residues in pork tissue. I. Assay by liquid chromatography with electrochemical detector

A liquid chromatographic (LC) method with electrochemical detection in the reductive mode was developed for the quantitative determination of dimetridazole (DMZ) and its major metabolite (HMMNI) at residue levels in pork tissue. For blood plasma, a sample is precipitated with 2 volumes of acetonitrile and centrifuged, and a diluted aliquot of the supernatant liquid is chromatographed. For muscle, a 10 g sample is extracted 3 times with dichloromethane. After evaporation of the combined extracts, the residue is redissolved in a mixture of hexane and mobile phase (0.3% TEA in 0.6M ammonium acetate pH 5.0 and acetonitrile, 85 + 15) and centrifuged, and an aliquot of the lower phase is chromatographed. Chromatography is accomplished using valve switching with 2 liquid circuits, employing the same mobile phase for both. The sample is deaerated by sparging with helium under slight positive pressure to prevent rediffusion of the oxygen. The sample is first loaded into a deoxygenator and the flow is stopped for complete deoxygenation. The flow is then resumed to transfer the sample into the first, low back-pressure column (ODS, 10 microns, 4.6 x 200 mm). Switching the valve at this point removes the deoxygenator from the circuit and connects the first column to a second one (ODS, 5 microns, 4.6 x 150 mm) in tandem. After the effluent is passed through a second deoxygenator to reduce the residual oxygen in the mobile phase, it is monitored by an electrochemical detector with a screened wall jet cell and a gold mercury electrode, set at -1.2 V.(ABSTRACT TRUNCATED AT 250 WORDS)     

Adsorption of Atrazine and Alachlor to Aquifer Material and Soil

Atrazine [6-chloro-N-ethyl-N′-(1-methyl)-1,3,5 triazine-2,3-diamine] and alachlor [2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl) acetamide] are agricultural herbicides used in large quantities and, as a consequence, are common contaminants in groundwater and surface water. The retention of these herbicides in soils and their degradation in aqueous environments is highly dependent upon their adsorption to solid surfaces. The adsorption of atrazine and alachlor was investigated on three typical Kansas and underlying aquifers known to be vulnerable to contamination. More alachlor was adsorbed to the soils and sediments than atrazine. The adsorption coefficients for atrazine were 2 to 5 times higher for soils than for aquifer sediments. For alachlor, the adsorption coefficients were 4 to 20 times higher for soil than for aquifer solids. Both linear and Freundlich isotherms represented the adsorption data well in all cases. The slope of the Freundlich isotherms, 1/n, was close to one, with the exception of alachlor adsorption onto the Topeka aquifer sediment (1/n = 0.67). The K _d values found in these studies were comparable to the lower range of those reported in the literature.     

泡沫法分离苜蓿叶蛋白工艺优化

为了研究泡沫分离苜蓿分离的条件,试验在单因素的基础上,采用响应面法对pH值、装液量、料液比、气体流速4个因素进行优化,以富集比和回收率为指标,最佳工艺条件：pH值为7.0、料液比为87.5 mg/L、装液量为600 mL、气体流速为600 mL/min。在此条件下叶蛋白的回收率为90.2%;富集比为7.64。泡沫分离法分离苜蓿蛋白是一种简单、有效的方法,该研究可为苜蓿叶蛋白的深加工提供参考。     

pH of the Pod-zone Affects Reproductive Growth of Groundnut

Most of the sandy soils that are suitable for production of groundnut (Arachis hypogaea L.) in the tropic and subtropics are acidic. Whilst effects of pH in the root zone have been studied, the effects of pod-zone pH on groundnut productivity remain relatively unknown. To develop appropriate soil management practices for groundnut production on acid soils, it is essential to understand how low pH affects reproductive growth of groundnut. Consequently, a glasshouse experiment was conducted in which attached groundnut gynophores were cultured in solution at pH ranging from 3.0 to 7.0. Low pH delayed pod initiation, and resulted in almost no pod expansion at pH 3.0. Only 12% and 55% of the cultured gynophores developed into pods at pH 3.0 and 4.0, respectively, compared with 91–95% at pH ≥ 5.0. Pods produced at pH 3.0 contained no seeds and those produced at pH 4.0 had a hollow, dark colored area in the cotyledon. Normal seeds and embryos were formed at pH ≥ 5.0, and plumule development was faster at solution pH ≥ 5.0 than at pH 4.0. Pod and kernel dry mass were optimised (90% of maximum) at pH 5.62–6.69 and 5.65–6.78, respectively. Septate and non-septate pod hairs were formed at all solution pH regimes, but were denser and more persistent at the higher pH. Kernel calcium (Ca) concentration decreased with decreasing pH, and was highly correlated with solution pH. Thus, pod-zone pH has important effects on the reproductive growth of groundnut, emphasizing the importance of managing pod-zone pH.     

Degradation of primisulfuron by a combination of chemical and microbiological processes

Microbial degradation of the herbicide primisulfuron was investigated using enrichment cultures from contaminated soils and 20 axenic cultures. At neutral pH, no disappearance of the herbicide was detected either in the enrichment cultures or in the growth media of the axenic microbial cultures. During the growth of some of the microbial strains, however, the pH of the medium dropped below 6, resulting in the hydrolysis of primisulfuron. The rate of primisulfuron hydrolysis was clearly pH dependent; primisulfuron was more persistent in neutral or weakly basic solutions than in acidic solutions. After hydrolysis of the herbicide, four products were observed. These were identified as methyl 2-(aminosulfonyl)benzoate, 2-amino-4,6-(difluoromethoxy)pyrimidine, 2-N-[[[[[4, 6-bis(difluoromethoxy)-2-pyrimidinyl]amino]carbonyl]amino]sulfonyl ]be nzoic acid, and 2-(aminosulfonyl)benzoic acid. After hydrolysis, it was found that the fungus Phanerochaete chrysosporium mineralized 27 and 24% of (14)C-phenyl- and (14)C-pyrimidine-labeled products, respectively, after 24 days of incubation. Similarly, Trametes versicolor mineralized 13 and 11% of (14)C-phenyl- and (14)C-pyrimidine-labeled hydrolysis products, respectively. In addition, primisulfuron in a hydrolytically stable solution, at pH 7. 0, was rapidly decomposed after ultraviolet irradiation, and two photolysis products were isolated [methylbenzoate and 4, 6-(difluoromethoxy)pyrimidin-2-ylurea]. When (14)C-phenyl-labeled primisulfuron was exposed to photolysis for 24 h, 32% of the initial radioactivity was recovered as (14)CO(2), whereas no (14)CO(2) was detected if the herbicide was labeled at the (14)C-pyrimidine position. Mineralization of (14)C-pyrimidine-labeled products of photolyzed primisulfuron by P. chrysosporium was approximately 25% after 24 days. These results clearly indicate that hydrolysis and photolysis of primisulfuron facilitated microbial degradation.     

High-performance capillary electrophoresis with indirect UV detection for determination of alpha-galactosides in Leguminosae and Brassicaceae

A rapid, easy, and reproducible capillary electrophoresis method for determination of raffinose family oligosaccharides (alpha-galactosides) was developed. Sucrose, raffinose, stachyose, verbascose, and ajugose were determined with indirect UV detection at moderate alkaline pH 9.2, using pyridine-2,6-dicarboxylic acid as background electrolyte in a sodium tetraborate buffer with added cetyltrimethylammonium bromide. The separation efficiency measured by the number of theoretical plates (N) ranged from 1.4 x 10(5) to 2.3 x 10(5). The precision of the method, measured by the relative standard deviation (RSD), was less than 0.53% for the migration times and better than 3.4% for normalized areas (NA), considering all sugars except verbascose (RSD(NA) = 11.8%). Detection limits were about 110 microg/mL, corresponding to 150-320 microM. Relative response factors (RRF) were calculated on the basis of linearity studies and used for quantification of alpha-galactosides in a lupine sample (Lupinus angustifolius).     

Separation and preconcentration of Cd(II), Cu(II), Ni(II), and Pb(II) in water and food samples using Amberlite XAD-2 functionalized with 3-(2-nitrophenyl)-1H-1,2,4-triazole-5(4H)-thione and determination by inductively coupled plasma-atomic emission spectrometry

A separation and preconcentration procedure was developed for the determination of trace amounts of Cd(II), Cu(II), Ni(II), and Pb(II) in water and food samples using Amberlite XAD-2 fuctionalized with a new chelating ligand, 3-(2-nitrophenyl)-1H-1,2,4-triazole-5(4H)-thione (Amberlite XAD-2-NPTT). The chelating resin was characterized by Fourier transform infrared spectroscopy (FT-IR) and used as a solid sorbent for enrichment of analytes from samples. The sorbed elements were subsequently eluted with 10 mL of 1.0 M HNO(3), and the eluates were analyzed by inductively coupled plasma-atomic emission spectrometry. The influences of the analytical parameters including pH, amount of adsorbent, eluent type and volume, flow rate of the sample solution, volume of the sample solution, and effect of matrix on the preconcentration of metal ions have been studied. The optimum pH for the sorption of four metal ions was about 6.0. The limits of detection were found to be 0.22, 0.18, 0.20, and 0.16 μg L(-1) for Cd(II), Cu(II), Ni(II), and Pb(II), respectively, with a preconcentration factor 60. The proposed method was applied successfully for the determination of metal ions in water and food samples.     

BIOLOG法测定土壤微生物群的一些局限性

Reduction of Cr(VI)to Cr(Ⅲ）were studied in a fresh wheat rhizosphere soil(Kuroboku,high humicandosol) pretreated with a basal fertilizer consisting of (NH4)2SO4,P2O5 and KH2PO4 and with K2Cr2O7 by using a rhizobox system.It was found that rhizosphere exerted a positive effect on Cr(VI) reduction.Part of the reason was the decrwease of pH in the rhizosphere due to application of (NH4)2SO4,implying that application of physiologically acid fertilizers would reduce Cr(VI) toxicity to plants.     

四川和重庆地区的植茶土壤与茶叶品质

A laboratory experiment was conducted to evaluate the effect of triphenyltetrazolum chloride(TTC) on soil microorganisms and the availability of pH characterization medium in BIOLOG plates.Application of TTC decreased the color development sharply and resulted in a great biocidal effect on the growth and reproduction of soil microorganisms,indicating that TTC can affect the discrimination on soil microbial community.The microtitration plates with 21 cabon sources and two different pH levels(4.7 and 7.0) were used to determine microbial community structure of eight red soils.The average utilization(average well colour development) of the carbon sources in the paltes with different pH levels generally followed the same sigmoidal pattern as that in the traditional BIOLOG plates,but the pH 4.7 plates increased the discrimination of this technique,compared with the pH 7.0 plates.Since most tested soils are acid,it seemed that it‘s better to use a suitable pH characterization medium for a specific spil in the sole carbon source test.     

中空纤维膜分离燕麦蛋白工艺及膜清洗方案

以燕麦蛋白回收率、总糖清除率、膜的污染度、浓缩效率及膜通量恢复率为指标,优化了中空纤维膜分离燕麦蛋白的工艺参数及膜的清洗方案。在温度35℃、压力0.02 MPa、pH值=8、流量25 L/h条件下利用MOF503分离经滤纸预处理的燕麦蛋白提取液,燕麦蛋白回收率、总糖清除率、膜的污染度及浓缩效率分别为80.3%、79.5%、41%、 43.5 L/(m2·h),同时,若采用分批超滤方式并辅以定期在线负压反冲清洗更有利于MOF503超滤处理燕麦蛋白提取液;通过物理和化学清洗方法对MOF503清洗效果的研究,表明物理清洗中的负压反冲清洗效果要明显优于等压水力清洗,化学清洗中HCl(pH值=2)和NaOH+H2O2+HCl比其他的效果要好,若物理和化学清洗协同作用,可使MOF503膜通量恢复率达94.74%。该研究表明利用中空纤维膜分离燕麦蛋白是可行的,并且将为膜分离蛋白的研究提供一定的技术依据。     

Determination of tri-n-butyltin and di-n-butyltin compounds in fish by gas chromatography with flame photometric detection

An analytical method is described for the simultaneous quantitative determination of tri-n-butyltin and di-n-butyltin compounds in fish. The sample was extracted with 0.5N HCl-methanol, and the methanol solution was extracted with hexane. The extract was purified by gel permeation chromatography and treated with Grignard reagent to yield the methyl derivatives, which were determined by gas chromatography with flame photometric detection operated in the tin mode (610 nm). Recoveries of tri-n-butyltin chloride (Bu3SnCl) and di-n-butyltin dichloride (Bu2SnCl2) spiked to fish at the levels of 0.2 and 1.0 ppm ranged from 80 to 105%. Detection limits were 0.02 micrograms/g for both compounds. Tri-n-butyltin compounds equivalent to Bu3SnCl levels of 0.07-2.0 ppm and di-n-butyltin compounds equivalent to Bu2SnCl2 levels of 0.02-0.11 ppm were found in reared yellowtails, and these values showed good agreement with the results from gas chromatographic-mass spectrometric analysis.     

产纤维素酶菌群的筛选及其酶学特性研究

从常年堆放的腐质豆秆中分离到1组产纤维素酶菌群MO,并在50℃、pH8.0条件下培养,90h时达到最高酶活,活性为1.3611IU。该酶最适反应温度为60℃,pH为7.0,在60℃以下和pH3~8范围内具有良好的热稳定性和pH稳定性。在最适反应条件下,该酶的最高活性可达2.13IU。通过生长动力学研究了菌群内部不同生长阶段pH、酶活和失重率的相互关系,并通过非变性电泳对酶谱进行初步研究,得到7条活性条带,说明MO中具有多种产酶菌株。     

Improvement in the determination of mancozeb residues by the carbon disulfide evolution method using flow injection analysis

The sample decomposition of the carbon disulfide evolution method for the determination of dithiocarbamate residues was carried out in a closed vial in the presence of hexane. The evolved carbon disulfide was extracted by the organic solvent and injected in a flow system for its quantification as copper complex. The conditions for batch decomposition, flow injection determination, and association of both were investigated with sodium diethyldithiocarbamate as model substance. An one-channel flow system was employed where the carrier stream was the ethanolic ethylenediamine/copper solution. The determination range was of 0. 01-1.26 microg of CS(2), with a relative standard deviation of 0.06% (n = 10), with a sample throughput of 45 samples/h. The association of the batch decomposition with the flow system was carried out with the fungicide mancozeb and was applied to the analysis of its residue in potato, lettuce, cucumber, and green bean crops. The approach allowed the analysis of 11 samples in triplicate in 2 h, with recoveries between 85% and 92% and relative standard deviation about 2%.     

Isolation of a selected microbial consortium from a pesticide-contaminated mix-load site soil capable of degrading the herbicides atrazine and alachlor

Contaminated soil from a 100-year-old mix-load site located in Reading, PA was evaluated for its potential to provide indigenous microorganisms capable of degrading two widely utilized herbicides, atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT) and alachlor (2-chloro-2′,6′-diethyl-N-[methoxymethyl]-acetanilide; AL). Three different locations from the site were chosen for experimentation based on herbicide handling activities. Standard enrichment techniques were used to isolate a selective microbial consortium (SCM) with the desirable degrading capabilities. Three enrichment treatment schemes were evaluated; AT and AL, AL alone, and only AT. Degradative organisms were isolated from only one of the sample locations. Considerable differences in the soil parameters of the three sample locations were found that might have had an effect on the ability of the indigenous microbial populations within the soil to degrade AT and AL. In the initial cultures from this location, degradation occurred in the AT and AL treatment only. Because the AT and AL were the only sources of carbon and nitrogen (N) for the microbes, these results suggest that AL alone was not a sufficient N source. In general, the ability to degrade AL by the SMC was dependent on AT degradation. Alachlor degradation did not begin until approximately 15% of the AT was transformed. Once all of the AT was removed very little further AL degradation occurred. The average half-life (t_1/2) of AT was 7.5 d, while average t_1/2 for AL degradation was 11 d. Individual colonies from the SMC were identified by fatty acids methyl ester (FAME) analysis. Five strains were identified with similarity indexes above 70%. These isolates included the following: Alcaligenes xylosoxydans subsp. denitrificans, Alcaligenes xylosoxydans subsp. xylosoxydans, Pseudomonas putida, Pseudomonas marginalis, and Providencia rustigianii.     

Mixture approach for optimizing lycopene extraction from tomato and tomato products

A simple mixture process design based on the comparison of both quadratic and special cubic models and involving three mixture components (hexane/acetone/ethanol) as a solution for extracting lycopene from raw tomato, tomato sauce, and tomato paste was used to confirm the hypothesis that lycopene extraction rates are a function of the solvent used during the extraction process. Conventional criteria (p 相似文献