Degradation of chloroacetanilide herbicides by anodic fenton treatment

Anodic Fenton treatment (AFT) is an electrochemical treatment employing the Fenton reaction for the generation of hydroxyl radicals, strong oxidants that can degrade organic compounds via hydrogen abstraction. AFT has potential use for the remediation of aqueous pesticide waste. The degradation rates of chloroacetanilides by AFT were investigated in this work, which demonstrates that AFT can be used to rapidly and completely remove chloroacetanilide herbicides from aqueous solutions. Acetochlor, alachlor, butachlor, metolachlor, and propachlor were treated by AFT, and parent compound concentrations were analyzed over the course of the treatment time. Degradation curves were plotted and fitted by the AFT kinetic model for each herbicide, and AFT model kinetic parameters were used to calculate degradation rate constants. The reactivity order of these five active ingredients toward hydroxyl radical was acetochlor approximately metolachlor > butachlor approximately alachlor > propachlor. Treatment of the chloroacetanilides by AFT removed the parent compounds but did not completely mineralize them. However, AFT did result in an increase in the biodegradability of chloroacetanilide aqueous solutions, as evidenced by an increase in the 5-day biochemical oxygen demand to chemical oxygen demand ratio (BOD5/COD) to >0.3, indicating completely biodegradable solutions. Several degradation products were formed and subsequently degraded, although not always completely. Some of these were identified by mass spectral analyses. Among the products, isomers of phenolic and carbonyl derivatives of parent compounds were common to each of the herbicides analyzed. More extensively oxidized products were not detected. Degradation pathways are proposed for each of the parent compounds and identified products.     

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。     

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.     

Degradation of cyhalofop-butyl (CyB) by Pseudomonas azotoformans strain QDZ-1 and cloning of a novel gene encoding CyB-hydrolyzing esterase

Cyhalofop-butyl (CyB) is a widely used aryloxyphenoxy propanoate (AOPP) herbicide for control of grasses in rice fields. Five CyB-degrading strains were isolated from rice field soil and identified as Agromyces sp., Stenotrophomonas sp., Aquamicrobium sp., Microbacterium sp., and Pseudomonas azotoformans; the results revealed high biodiversity of CyB-degrading bacteria in rice soil. One strain, P. azotoformans QDZ-1, degraded 84.5% of 100 mg L(-1) CyB in 5 days of incubation in a flask and utilized CyB as carbon source for growth. Strain QDZ-1 could also degrade a wide range of other AOPP herbicides. An esterase gene, chbH, which hydrolyzes CyB to cyhalofop acid (CyA), was cloned from strain QDZ-1 and functionally expressed. A chbH-disrupted mutant dchbH was constructed by insertion mutation. Mutant dchbH could not degrade and utilize CyB, suggesting that chbH was the only esterase gene responsible for CyB degradation in strain QDZ-1. ChbH hydrolyzed all AOPP herbicides tested as well as permethrin. The catalytic efficiency of ChbH toward different AOPP herbicides followed the order quizalofop-P-ethyl ≈ fenoxaprop-P-ethyl > CyB ≈ fluazifop-P-butyl > diclofop-methyl ≈ haloxyfop-P-methyl; the results indicated that the chain length of the alcohol moiety strongly affected the biodegradability of the AOPP herbicides, whereas the substitutions in the aromatic ring had only a slight influence.     

Hapten design in the development of competitive enyme-linked immunosorbent assays for genotoxic metabolites of alachlor.

The acetanilide compounds 2-chloro-2',6'-diethylacetanilide (CDA) and 2-hydroxy-2',6'-diethylacetanilide (HDA) are environmental degradative products of the chloroacetanilide herbicide alachlor. CDA, HDA, and alachlor are ground and surface water contaminants. CDA and HDA are genotoxic in bacterial and mammalian test systems. This paper reports hapten design in the development of two competitive enzyme-linked immunosorbent assays (cELISA) for the detection of CDA and HDA. Chloroacetanilide herbicides and other alachlor metabolites that may be present in environmental samples do not cross-react with the detection of CDA and HDA. Solid-phase extraction of CDA and HDA residues from aqueous samples results in a 1000-fold concentration factor, resulting in an effective detection limit of 15 pg/mL for both assays. The specificity of the cELISAs required preservation of the degree of substitution of the acetanilide moiety in the hapten design. The hapten synthesis strategies are suitable for metabolites of other chloroacetanilide herbicides (i.e., acetochlor, butachlor, metolachlor, and propachlor).     

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.     

Structural influences in relative sorptivity of chloroacetanilide herbicides on soil

Adsorption of the chloroacetanilide herbicides acetochlor, alachlor, metolachlor, and propachlor was determined on soils and soil components, and their structural differences were used to explain their sorptivity orders. On all soils and soil humic acids, adsorption decreased in the order: metolachlor > acetochlor > propachlor > alachlor. On Ca(2+)-saturated montmorillonite, the order changed to metolachlor > acetochlor > alachlor > propachlor. FT-IR differential spectra of herbicide-clay or herbicide-humic acid-clay showed possible formation of hydrogen bonds and charge-transfer bonds between herbicides and adsorbents. The different substitutions and their spatial arrangement in the herbicide molecule were found to affect the relative sorptivity of these herbicides by influencing the reactivity of functional groups participating in these bond interactions. It was further suggested that structural characteristics of pesticides from the same class could be used to improve prediction of pesticide adsorption on soil.     

Persistence of Atrazine and Alachlor in Ground Water Aquifers and Soil

Degradation of atrazine and alachlor in saturated aquifer materials and soil was studied in the laboratory. A static aquifer was represented by a set of stagnant flasks and a well-mixed aquifer was simulated by recirculating columns. Water was tested at selected time intervals over six months and analyzed for herbicides and metabolites. Under all conditions, atrazine was more persistent than alachlor. Increased temperature had little effect on atrazine dissipation but did increase alachlor degradation rates, especially in the sterilized treatments. The addition of carbon and nitrogen prolonged the initial period before the onset of degradation in some of the columns. Enhanced mass transfer of the herbicides, nutrients, and oxygen in the recirculating columns dramatically increased dissipation of atrazine and alachlor. The degradation rates of atrazine and alachlor were 2 to 5 times faster in the recirculating columns than in the stagnant flasks. Atrazine was more persistent in the aquifer materials than in the soils, while alachlor dissipation was similar in the soils and recirculating aquifer columns, but was slower in the stagnant flasks. The prolonged persistence of atrazine under static, aquifer conditions (t _1/2 = 206 to 710 days) indicates that natural mechanisms are not sufficient to alleviate the risk of atrazine buildup over time; however, in a well mixed aquifer, atrazine degradation rates should be higher (t _1/2 = 66 to 106 days) and the threat of atrazine accumulation is diminished. Alachlor persistence at low concentrations (< 10 μg L^?1) in aquifers should not pose a long-term threat to ground water supplies.     

丁草胺和苄嘧磺隆对钝顶螺旋藻的毒性效应研究

采用实验室摇瓶实验法研究了除草剂丁草胺和苄嘧磺隆对钝顶螺旋藻的毒性效应。结果表明,丁草胺浓度与藻生长（以OD560计）存在明显的剂量-效应关系,64mg·L^-1丁草胺严重抑制藻生长,比生长速率为0.111d-1,仅为对照的24%。在7d实验时间内EC50值随染毒时间延长呈指数下降,96h的EC50值为45.4mg·L^-1,到第7d降为15.2mg·L^-1。与丁草胺相似,在实验浓度范围内随浓度增加,苄嘧磺隆对钝顶螺旋藻的生长抑制作用逐渐增强,最高浓度44.8mg·L^-1处理的比生长速率为0.458d-1,约为对照的82%。在8d实验时间内EC50值随染毒时间延长呈指数增长,96h的EC50值为51.3mg·L^-1,到第8d达到215mg·L^-1。两种除草剂在较高浓度条件下均使钝顶螺旋藻藻丝变短,部分藻丝由螺旋型变为直线型,随浓度增加直线型藻丝逐渐增加,到最高浓度几乎全部变为直线型,且藻细胞颜色变浅。依据96h-EC50值判断丁草胺对钝顶螺旋藻的毒性大于苄嘧磺隆,二者毒性强度均为中等,但随着染毒时间延长丁草胺的毒性作用增强而苄嘧磺隆的毒性逐渐降低。同时,高浓度的两种除草剂使部分藻丝断裂变短,且诱导藻丝形态由螺旋型向直线型转变。     

除草剂对土壤温室气体排放的影响

试验设对照、尿素、尿素＋草甘膦和尿素＋丁草胺4个处理,尿素氮用量为200mg·kg-1干土,除草剂用量为10mg有效成分·kg-1干土。在实验室恒温培养条件下,研究除草剂对菜田土壤温室气体排放的影响。结果表明,菜田土壤中施用氮肥显著增加了温室气体N2O、CO2和CH4的排放。尿素氮肥中添加草甘膦显著抑制N2O、CO2的排放,分别比尿素处理降低48.4%和20.2%;添加丁草胺显著抑制N2O排放,比尿素处理降低23.2%,对CO2排放略有减少但不显著;草甘膦和丁草胺对CH4排放都无明显影响。这说明除草剂对土壤温室气体的排放具有显著影响,但不同除草剂品种的效应也存在明显差异。因此,在农田温室气体排放估算时应考虑除草剂的施用对温室气体减排所产生的效果。     

Acute Toxicity of Seven Selected Pesticides (Alachlor, Atrazine, Dieldrin, Diuron, Pirimiphos-Methyl, Chlorpyrifos, Diazinon) to the Marine Fish (Turbot, Psetta maxima)

The present study evaluated the short-term toxicity of seven selected pesticides: four insecticides (chlorpyrifos, dieldrin, diazinon and pirimiphos-methyl) and three herbicides (diuron, alachlor and atrazine). With this aim, a standard toxicity test with the highly sensitive early life stages (ELS) of a marine fish was used. The turbot, Psetta maxima, is abundant in shallow estuarine and costal habitats and is currently the most commonly cultivated fish species in Galicia, NW Spain. According to the turbot ELS test results, chlorpyrifos was the most toxic pesticide tested for both embryos and larvae and was followed in order of decreasing toxicity by dieldrin, pirimiphos-methyl, diazinon, alachlor, atrazine and diuron. Larvae were more sensitive than embryos to the seven pesticides. The median lethal concentrations of the selected pesticides during a 48- and a 96-h exposure for turbot embryos and larvae were, respectively (in micrograms per litre): chlorpyrifos, 116.6 and 94.65; dieldrin, 146 and 97; pirimiphos-methyl, 560 and 452; diazinon, 1,837 and 1,230; alachlor, 2,177 and 2,233; diuron, 10,076 and 7,826; and atrazine, 11,873 and 9,957. According to their acute toxicity, the insecticides were more toxic than the herbicides. Furthermore, all insecticides and herbicides appear to be teratogenic to turbot ELS.     

Influence of herbicide structure,clay acidity,and humic acid coating on acetanilide herbicide adsorption on homoionic clays

Adsorption of chloroacetanilide herbicides on homoionic montmorillonite was studied by coupling batch equilibration and FT-IR analysis. Adsorption decreased in the order metolachlor > acetochlor > alachlor > propachlor on Ca(2+)- or Mg(2+)-saturated clays and in the order metolachlor > alachlor > acetachlor > propachlor on Al(3+)- or Fe(3+)-saturated clays. FT-IR spectra showed that the carbonyl group of the herbicide molecule was involved in bonding. For the same herbicide, adsorption of alachlor, acetachlor, and metolachlor on clay followed the order Ca(2+) approximately Mg(2+) < Al(3+) < or = Fe(3+), which coincided with the increasing acidity of homoionic clays. Adsorption of propachlor, however, showed an opposite dependence, suggesting a different governing interaction. In clay and humic acid mixtures, herbicide adsorption was less than that expected from independent additive adsorption by the individual constituents, and the deviation was dependent on the clay-to-humic acid ratio, with the greatest deviation consistently occurring at a 60:40 clay-to-humic acid ratio.     

活性污泥中细菌对聚丙烯酰胺的生物降解研究

聚丙烯酰胺（Polyacrylamide,PAM）是一类重要的水溶性高分子聚合物,已广泛应用到工农业生产的各个领域和人们的日常生活中。同时,PAM在环境中的残留、迁移和降解对环境的污染也日趋严重,尤其是降解后的单体丙烯酰胺对人类的神经系统有很大的危害。本文从胜利油田的活性污泥中筛选出3株聚丙烯酰胺降解菌,通过比较筛选出一株降解效果较好的菌,命名为AS－2。根据生理生化特性分析,初步鉴定为海球菌属。采用室内培养方法,研究了AS－2对聚丙烯酰胺生物降解的最佳条件。结果表明,当降解时间为5d,pH=8,温度为40℃,碳源为原油,氮源为NaNO3,原油和NaNO3的含量分别为2．5,1．4g·L^-1时,AS-2对聚丙烯酰胺的降解率达到45．23％。通过对聚丙烯酰胺生物降解前后的红外谱图比较,推断出AS-2主要降解了聚丙烯酰胺侧链的酰胺基,将酰胺基降解为羧酸和游离的氡基。用高效液相色谱检测生化后的PAM溶液,未检测出单体丙烯酰胺。     

乙草胺降解菌Y-4的分离鉴定及降解特性研究

从长期经乙草胺污染的污泥中分离到一株能以乙草胺为唯一碳源和能源生长的菌株Y-4,通过生理生化实验和16S rDNA同源性序列分析,鉴定为申氏杆菌属（Shinella sp.）。采用室内培养方法,研究了Y-4对乙草胺的降解特性。结果表明,Y-4能有效地降解浓度为5～200 mg.L-1的乙草胺,在48 h内对50 mg.L-1乙草胺的降解率达到83.3%。菌株Y-4降解乙草胺的最适pH值为8.0,最适温度为30℃,其对丙草胺和丁草胺等农药也有良好的降解效果。     

Determination of alachlor and its metabolite 2,6-diethylaniline in microbial culture medium using online microdialysis enriched-sampling coupled to high-performance liquid chromatography

In this study, a simple and novel microdialysis sampling technique incorporating hollow fiber liquid phase microextraction (HF-LPME) coupled online to high-performance liquid chromatography (HPLC) for the one-step sample pretreatment and direct determination of alachlor (2-chloro-2',6'-diethyl-N -(methoxymethyl)acetanilide) and its metabolite 2,6-diethylaniline (2,6-DEA) in microbial culture medium has been developed. A reversed-phase C-18 column was utilized to separate alachlor and 2,6-DEA from other species using an acetonitrile/water mixture (1:1) containing 0.1 M phosphate buffer solution at pH 7.0 as the mobile phase. Detection was carried out with a UV detector operated at 210 nm. Parameters that influenced the enrichment efficiency of online HF-LPME sampling, including the length of the hollow fiber, the perfusion solvent and its flow rate, the pH, and the salt added in sample solution, as well as chromatographic conditions were thoroughly optimized. Under optimal conditions, excellent enrichment efficiency was achieved by the microdialysis of a sample solution (pH 7.0) using hexane as perfusate at the flow rate of 4 μL/min. Detection limits were 72 and 14 ng/mL for alachlor and 2,6-DEA, respectively. The enrichment factors were 403 and 386 (RSD < 5%) for alachlor and 2,6-DEA, respectively, when extraction was performed by using a 40 cm regenerated cellulose hollow fiber and hexane as perfusion solvent at the flow rate of 0.1 μL/min. The proposed method provides a sensitive, flexible, fast, and eco-friendly procedure to enrich and determine alachlor and its metabolite (2,6-DEA) in microbial culture medium.     

Acid- and base-catalyzed hydrolysis of chloroacetamide herbicides

Despite the prevalence of chloroacetamides as herbicides, little is known about the rates or products of acid- or base-catalyzed hydrolysis of these compounds. Mechanisms of acid-catalyzed reactions may parallel those catalyzed by (hydr)oxide minerals, while base-catalyzed processes have as important counterparts reactions with environmental nucleophiles (such as reduced sulfur species). The current study systematically investigates how the structure of nine chloroacetamides affects their reactivity in 2 N NaOH, 2 N HCl, or 6 N HCl at 25 or 85 degrees C. Base-catalyzed hydrolysis proceeds either through an intermolecular SN2 reaction to hydroxy-substituted derivatives or (in a few cases) through amide cleavage, while both amide and ether group cleavages are observed under acidic conditions. Our results reveal that subtle differences in chloroacetamide structure [notably the type of (alkoxy)alkyl substituent] can dramatically influence reactivity and reaction mechanism. Hydroxy-substituted, morpholinone, and secondary aniline derivatives were identified upon reaction for several years in deionized water at circumneutral pH.     

Biodegradation of aged residues of atrazine and alachlor in a mix-load site soil

A selected microbial consortium (SMC) capable of degrading two specific herbicides, alachlor (2-chloro-2′,6′-diethyl-N-[methoxymethyl]-acetanilide; AL) and atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT) was isolated from a pesticide-contaminated mix-load site soil. Evaluation of bioaugmentation as a feasible bioremediation strategy for this mix-load site soil (Site 5A) was initiated in standard laboratory biometer flasks utilizing the isolated SMC. The biometer flasks were monitored for CO₂ evolution and pesticide degradation. The total amount of CO₂ evolved from the treated biometer flasks was significantly different from the control flasks. The rate of CO₂ evolution was 2.6 times faster in the treated soil (0.0123 mM CO₂ d⁻¹ vs. 0.0048 mM CO₂ d⁻¹). The total net CO₂ produced in the treated biometer flasks was 0.9481 mM, representing mineralization of approximately 10% of the AT and AL initially present. Forty-eight percent of AT and 70% of AL was degraded in the inoculated biometer flasks. The first-order rate constants were 0.0064 d⁻¹ and 0.1331 d⁻¹ for AT and AL, respectively. The calculated half-life of AT was 108 d while a 50% decrease in AL occurred by Day 5. In just 2 d, 20% of the AT was degraded while only 10% of the AL disappeared. The initial fast degradation rate of AT was followed by a much slower, more gradual degradation rate period that lasted about 35 d. Alternatively, the rate of AL degradation increased after the second day resulting in 60% of the AL being transformed by the end of the first week. Alachlor degradation appeared to be dependent upon AT degradation especially during the first several days of the incubation period. Complete disappearance of the herbicides over the study time was not achieved.     

除草剂对不同种植年限柑橘园土壤氮转化过程及温室气体排放的影响

为探讨除草剂施用对柑橘园土壤氮转化及温室气体排放的影响,在实验室培养条件下,研究了0年(林地)、种植10年和30年的柑橘园土壤中分别添加除草剂草甘膦和丁草胺后,尿素态氮含量、硝化和反硝化作用以及温室气体排放的变化。研究结果表明,橘园土壤中尿素第1 d的水解率、氮肥硝化率、反硝化作用损失总量以及N_2O和CO_2排放量显著高于林地土壤(P0.05)。与10年橘园土壤相比,30年橘园土壤显著增加了尿素的水解速率、氮肥硝化率和CO_2排放量(P0.05),但二者的反硝化损失量没有显著差异。施用草甘膦和丁草胺都显著促进了林地土壤的尿素水解(P0.05),第1 d尿素态氮含量分别降低11.20%和12.43%;但对3种土壤氮肥的硝化率均没有明显影响。施用丁草胺显著降低了林地土壤的CO_2排放量(P0.05),对两种橘园土壤的CO_2排放没有明显影响,但明显增加了两种橘园土壤的N_2O排放总量(P0.05),分别比不施除草剂增加56.27%和85.41%;施用草甘膦对3种土壤的N_2O和CO_2排放均没有明显影响。可见,草甘膦和丁草胺的施用不会对柑橘园土壤的氮转化过程产生影响,但丁草胺显著增加了柑橘园土壤的N_2O排放。     

Discriminating multiple impacts of biogas residues amendment in selectively decontaminating chloroacetanilide herbicides

There is increasing concern about modifications to pesticide persistence in soil from the application of organic wastes as fertilizers. This study was conducted to discriminate the multiple effects of biogas residues (BR) amendment, including soil nutrients, soil microbial activity and biodiversity, and adsorption and degradation of chloroacetanilide herbicides (acetochlor, metolachlor, and butachlor). Addition of BR to soil increased the release of organic materials (i.e., dissolved organic carbon, dissolved organic nitrogen, and active phosphorus). It not only stimulated soil microorganisms and caused changes to microorganism diversity but also increased herbicide adsorption. Such multiple effects led to selective decontamination of chloroacetanilide herbicides, depending on herbicide structures and BR amendment levels. Stereoselectivity in degradation of acetochlor and metolachlor with biphasic character was magnified by BR amendment, which was well explained by integrating the impacts of BR amendment. Interestingly, BR amendment induced significant accumulation of herbicidally active aS,CS-metolachlor, facilitating the utilization of herbicidal activity.