Cleavage of the diketonitrile derivative of the herbicide isoxaflutole by extracellular fungal oxidases

Isoxaflutole is a herbicide activated in soils and plants to its diketonitrile derivative, the active herbicide principle. The diketonitrile derivative undergoes cleavage to the inactive benzoic acid analogue. In this paper, it is established that an oxidative mechanism implicating two successive reactions in the presence of dimethyldioxirane can chemically initiate the cleavage of the diketonitrile. It is also shown that two white rot strains, Phanerochaete chrysosporium and Trametes versicolor, are able to convert the diketonitrile to the acid when cultured in liquid media. This main metabolite amounts to 24.6 and 15.1% of initial herbicide content after 12-15 days of culture. Another polar metabolite represents <3.7% of the parent compound amount during the same period. Oxidative enzymes produced by the fungi show a time course similar to that of diketonitrile degradation. Purified laccase (EC 1. 10.3.2), in the presence of 2 mM 2, 2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) acting as a redox mediator at pH 3 supports the reaction with rates of 0.3-0.4 nmol h(-)(1) unit(-)(1).     

Effect of soil properties on the degradation of isoxaflutole and the sorption-desorption of isoxaflutole and its diketonitrile degradate

The transformation of isoxaflutole (ISOX) to its herbicidally active diketonitrile degradate (DKN) was significantly enhanced in the presence of soil and occurred more rapidly in systems containing soil with a greater soil pH. Sorption-desorption of ISOX and DKN in five soils collected within a field revealed both ISOX and DKN were more readily sorbed to soils with greater organic matter, clay content, and lower soil pH. Sorption of ISOX residues occurred within 2 h, and extracts contained similar concentrations of ISOX and DKN at 24 h, suggesting the 24-h sorption coefficients for ISOX-treated systems were actually for mixed ISOX residues. Freundlich sorption coefficients were 3 and 4 times greater for ISOX than for DKN. On the basis of the Freundlich organic carbon sorption constants, ISOX and DKN can be categorized in the very high and high mobility classes, suggesting their potential to leach in the soils needs to be evaluated.     

Reaction pathways of the diketonitrile degradate of isoxaflutole with hypochlorite in water

Isoxaflutole (IXF; Balance) belongs to a new class of isoxazole herbicides. Isoxaflutole has a very short half-life in soil and rapidly degrades to a stable and phytotoxic degradate, diketonitrile (DKN). DKN was previously discovered to rapidly react with hypochlorite (OCl-) in tap water, yielding the benzoic acid (BA) degradate as a major product, but the complete reaction pathway and mechanism have not been elucidated. Thus, the objectives of this work were to (1) determine the stoichiometry of the reaction between DKN and OCl-; (2) identify products in addition to BA; and (3) propose a complete pathway and reaction mechanism for oxidation of DKN by OCl-. Stoichiometry of the reaction showed a molar ratio of OCl-/DKN of 2. In addition, two previously uncharacterized chlorinated intermediates were identified under conditions in which OCl- was the limiting reactant. The proposed chemical structure of a chlorinated benzoyl intermediate was inferred from a series of HPLC/MS and HPLC/MS/MS experiments and the use of mass spectral simulation software. A chlorinated ketone intermediate was also identified using ion trap GC/MS. Two additional end products were also identified: cyclopropanecarboxylic acid (CPCA) and dichloroacetonitrile (DCAN). On the basis of the reaction stoichiometry, the structure of the chlorinated intermediates, and the identification of the products, two reaction pathways are proposed. Both pathways involve a two-step nucleophilic attack and oxidation of the diketone structure of DKN, leading to formation of BA, DCAN, and CPCA.     

Fate of isoxaflutole in soil under controlled conditions

Isoxaflutole (IFT, 5-cyclopropyl-1,2-oxazol-4yl-alpha,alpha,alpha-trifluoro-2-mesyl-p-tolyl ketone) is a new pre-emergence proherbicide used in maize and sugarcane. Its two main derivatives are a diketonitrile derivative, 2-cyano-3-cyclopropyl-1-(2-methanesulfonyl-4-trifluoromethylphenyl)propane-1,3-dione, called DKN, and a benzoic acid derivative, 2-methanesulfonyl-4-trifluoromethylbenzoic acid, called BA. Few data are available of the factors influencing the degradation of IFT in soil, and the purpose of the present work was to determine the relative importance of, and factors affecting, the degradation of IFT in soil. Experiments were conducted on five soils with distinct physicochemical characteristics, at different temperatures and moisture contents in biotic and abiotic conditions. The isomerization of IFT to DKN is rapid, increasing with higher moisture contents and higher temperatures. It depends strongly on pH and is governed by chemical processes. The degradation of DKN to BA appeared to be essentially due to the biological activity of the soil.     

Mobility and degradation of the herbicide imazosulfuron in lysimeters under field conditions

The mobility and degradation of imazosulfuron, labeled with carbon-14 at the imidazole ([imi-(14)C]imazo) or pyrimidine ring ([pyr-(14)C]imazo), in lysimeters with 1 m(2) surface and 110 cm depth were investigated for three years. One lysimeter was treated with [imi-(14)C]imazo in two successive years at the rate of 50 g of active ingredient (ai)/ha each. The other two lysimeters were treated once with [pyr-(14)C]imazo and a mixture (1:1, w/w) of the two labeled imazosulfurons, respectively (50 g of ai/ha). In the first and second years of monitoring, the yearly mean concentration of (14)C in the leachate water was <0.10 microg/L in each lysimeter. Although in the third year the concentration of (14)C in the leachate water was 0.17 microg/L for the lysimeter treated twice with [imi-(14)C]imazo, the concentration of imazosulfuron and its degradation products in the leachate water ranged from 0.01 to 0.06 microg/L. At the study termination, the main portion of (14)C recovered was found in the upper 30 cm soil layer in each lysimeter, and no (14)C was detected below a depth of 50 cm. These findings indicated that imazosulfuron and its degradation products in soils translocated into groundwater only slightly.     

Recolonization of methyl bromide sterilized soils under four different field conditions

Summary The course of recovery in biological activity was assessed in the top 5 cm of undisturbed soil cores (29.7 cm diameter, 30 cm deep) that had been fumigated in the laboratory with methyl bromide. The cores were returned to their original pasture and forest sites, two with a moderate and two with a high rainfall, and untreated soils at all sites served as baselines. Sampling took place over 166 days (midsummer to midwinter). Microbial biomass (as measured by fumigation-extraction and substrate-induced respiration procedures) and dehydrogenase activity both recovered rapidly, but remained consistently lower in the fumigated than in untreated samples at both forest sites and at the moister of the two pasture sites. Bacterial numbers also recovered rapidly. Fungal hyphal lengths were, on average over 166 days, 25% lower in the fumigated soils. Levels of mineral N were initially highest in the fumigated soils, but declined with time. Fumigation generally had no detectable effects on the subsequent rates of net N mineralization and little effect on nitrification rates. Fumigation almost totally eliminated protozoa, with one to three species being recovered on day 0; the numbers recovered most rapidly in the moist forest soil and slowly in the dry pasture soil. The recoionization rate of protozoan species was similar in all soils, with species numbers on day 110 being 33 and 34 in the fumigated and untreated soils, respectively. Nematodes were eliminated by fumigation; recolonization was first detected on day 26 but by day 166, nematode numbers were still lower in fumigated than in untreated soils, the abundance being 10 and 62 g^-1 soil and diversity 10 and 31 species, respectively. Overall, the results suggest that protozoan and nematode populations and diversities could provide a useful medium-term ecological index of the recovery in comprehensive soil biological activity following major soil pollution or disturbance.     

Assessment of the leaching potential of 12 substituted phenylurea herbicides in two agricultural soils under laboratory conditions

In this study, the potential groundwater pollution of 12 substituted phenylurea herbicides (chlorbromuron, chlorotoluron, diuron, fenuron, fluometuron, isoproturon, linuron, metobromuron, metoxuron, monolinuron, Monuron, and neburon) was investigated under laboratory conditions. For this purpose, leaching studies were conducted using disturbed soil columns filled with two different agricultural soils, one hypercalcic calcisol (HC) and the other endoleptic phaeozem (EP). In the case of the HC, all of the studied herbicides were found in leachates, while for the EP only, chlorbromuron, chlorotoluron, isoproturon, monolinuron, and, especially, fenuron were recovered. For both soils, the groundwater ubiquity score (GUS) index was calculated for each herbicide on the basis of its persistence (as t(1/2)) and mobility (as K(OC)). The half-lives obtained were markedly higher in the EP (217-518 days) than in the HC (71-178 days). As a consequence, higher values of GUS indexes were observed for EP. The ratio of the GUS between the EP and the HC was about 1.3.     

小麦和土壤中环丙唑醇残留消解动态及膳食摄入评估

为评价环丙唑醇在小麦生产上应用的安全性,通过建立乙腈提取、氮磷检测器检测方法对小麦籽粒、植株和土壤样品中环丙唑醇的残留量进行检测,研究了小麦籽粒、植株和土壤中环丙唑醇的残留及其消解动态,并对小麦中的残留量进行风险评估。结果表明,环丙唑醇在小麦籽粒、植株及土壤空白样品中的添加回收率为79.2%~95.6%,相对标准偏差为1.9%~10.0%,最小检出量为8.2×10?12 g,在小麦籽粒、植株及土壤中的最低检测浓度均为0.05 mg·kg?1,乙腈提取、氮磷检测器检测方法重现性好,准确度、精密度高,可满足环丙唑醇在小麦上的残留分析要求。2010年和2011年,河南省、黑龙江省和江苏省3地环丙唑醇在小麦植株和土壤中的消解半衰期分别为3.0~5.5 d、18.1~34.5 d;不同施药次数、施药量及采样间隔,环丙唑醇在小麦籽粒中的最终残留量均≤0.415 mg·kg?1。采收间隔期为14 d和21 d时,不同施药次数、施药剂量和采收间隔期,环丙唑醇在小麦植株、小麦籽粒和土壤中的残留量差异均不显著;采收间隔期为7 d,有效成分108 g?hm-2施药2次与有效成分162 g?hm-2施药3次时小麦植株、小麦籽粒和土壤中的残留量之间均存在显著性差异。普通人群环丙唑醇的国家估算每日摄入量为0.000 109 9 mg,占日允许摄入量的0.5%左右,按本试验方式进行施药,通常不会对一般人群健康产生不可接受的风险。     

Estimation of the total gaseous nitrogen losses from clay soils under laboratory and field conditions

Acetylene blockage was evaluated as a method for measuring losses of N₂O + N₂ from two Denchworth series clay soils. The denitrification potential in anaerobic, dark incubations at 20°C with nitrate (equivalent to 100 kg N ha^?1 0–20 cm depth), maximum water holding capacity, and acetylene (1%), was equivalent to 32 ± 11 and 39 ± 6 kg N ha^?1 per day for the two 0–20 cm soils and was positively correlated with carbon content (r= 0.98). After 4 days N₂O was reduced to N₂ in the presence of C₂H₂. In April 1980 following irrigation (24 mm) and applications of ammonium nitrate (70 kg N ha^?1) and acetylene, the mean nitrous oxide flux from soil under permanent grass was 0.05 ± 0.01 kg N₂O-N ha^?1 per day for 8 days. In June 1980, the losses of nitrogen from cultivated soils under winter wheat after irrigation (36 mm) and acetylene treatment were 0.006 ± 0.002 and 0.04–0.07 ± 0.01 kg N ha^?1 per day respectively before and after fertilizer application (70 kg N ha^?1). The nitrous oxide flux in the presence of acetylene decreased briefly, indicating that nitrification was rate determining in drying soil.     

Permethrin and its two metabolite residues in seven agricultural crops

Metabolite residues of permethrin are not reported in the literature for most agricultural crops. This paper reports residues of permethrin and its 2 metabolites (dichlorovinyl acid and metaphenoxybenzyl alcohol) in 7 different agricultural crops (Chinese cabbage, spinach, asparagus, raspberries, green peas, turnip roots, and turnip greens). Permethrin residues declined approximately 85% within 7 days after treatment in all crops. In most cases, the acid metabolite residues peaked at 3 days, and declined after that. Translocation of residues into turnip roots was very slight; the average was less than 0.05 ppm for permethrin and alcohol metabolite residues and none was detected for the acid metabolite residue. Permethrin residues in the turnip greens averaged approximately 2 ppm for the 0.112 kg ai/ha treatment, and 6 ppm for the 0.224 kg ai/ha treatment.     

Enantioselective environmental behavior of the chiral herbicide fenoxaprop-ethyl and its chiral metabolite fenoxaprop in soil

The enantioselective degradation behavior of fenoxaprop-ethyl (FE) and its chiral metabolite fenoxaprop (FA) in three soils under native conditions was investigated. Two pairs of enantiomers were analyzed by high-performance liquid chromatography (HPLC) with an amylose tri-(3,5-dimethylphenylcarbamate) (ADMPC) chiral column. The degradation of racemic FE in three soils showed the herbicidally inactive S-(-)-enantiomer degraded faster than the active R-(+)-enantiomer. FE was configurationally stable in soils because no interconversion to the respective antipodes was observed during incubation of the enantiopure S-(-)- or R-(+)-FE. The main metabolites of FE were confirmed as FA and 6-chloro-2,3-dihydrobenzoxazol-2-one (CDHB), and the formation of the chiral metabolite FA showed enantioselectivity in soils. The degradation of rac-FA was also enantioselective with the S-(-)-FA preferentially degraded: the half-life (t(1/2)) of the S-form in the three soils ranged from 2.03 to 5.17 days, and that of R-form ranged from 2.42 to 20.39 days. The inversion of the S-(-)-enantiomer into the R-(+)-enantiomer occurred in two of the three soils when the enantiopure S-(-)- and R-(+)-FA were incubated. The data from sterilized control experiments indicated that the enantioselectivity of FE and FA was attributed to microbially mediated processes.     

Comparison of nitrogen mineralized under anaerobic and aerobic conditions for some agricultural and forest soils of washington

Abstract

Seven agricultural soils and eight forest soils from Washington state were tested for mineralizable nitrogen using both anaerobic and aerobic incubation procedures. Each procedure had been used previously to. develop nitrogen indices for agricultural and forested ecosystems. Forest soils mineralized less nitrogen under anaerobic than aerobic conditions, while the opposite was true for agricultural soils. There were statistically significant correlations between the two methods for each of the time periods tested. Experimental variations were consistently lower than previously reported.     

Liquid chromatographic determination of fluridone aquatic herbicide and its metabolite in fish and crayfish

A residue method is described for determination of the aquatic herbicide fluridone (1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone) and its metabolite (1-methyl-3-(4-hydroxyphenyl)-5-[3-(trifluoromethyl) phenyl]-4(1H)-pyridinone) in fish and crayfish tissues. Both compounds are extracted from tissues with methanol, and the extracts are subjected to acidic hydrolysis to release conjugated forms of fluridone and the metabolite. Sample extracts are purified by liquid-liquid partitioning and Florisil Sep-Pak column chromatography. Both compounds are separated and measured by reverse phase liquid chromatography with UV detection at 313 nm. In the absence of interfering peaks, the method has a detection limit of approximately 0.04 ppm of either compound. Overall, recoveries averaged 96% for fluridone and 78% for the metabolite for all tissue types combined.     

不同耕作方式下旱作玉米田土壤呼吸及其影响因素

为揭示不同耕作方式对旱作玉米田土壤呼吸的影响,对比研究深松耕、免耕、旋耕和翻耕4种耕作方式下土壤呼吸速率的动态变化及其与土壤水分、温度、有机质、全氮、pH值等的关系。结果表明,夏玉米生长季,4种耕作方式下土壤呼吸速率随生育时期均呈先增加后降低的趋势,平均土壤呼吸速率为深松耕>翻耕>旋耕>免耕;播种前至拔节期土壤温度为翻耕>深松耕>旋耕>免耕,抽雄期至成熟收获期为免耕>旋耕>深松耕>翻耕;各耕作方式下0～20cm层土壤有机质、全氮均逐渐增加,与免耕比较,翻耕有机质和全氮均降低;生育前期土壤pH值波动明显,抽雄期后趋于平缓,土壤pH值平均值为翻耕>旋耕>免耕>深松耕。各影响因素与土壤呼吸速率相关分析表明,深松耕和翻耕土壤水分、温度与土壤呼吸速率呈显著或极显著正相关;有机质与土壤呼吸速率呈负相关,且与深松耕措施下土壤呼吸速率呈显著负相关;除免耕外,其他耕作方式下土壤全氮、pH值与土壤呼吸呈负相关。该研究可为补充完善土壤呼吸排放机理、评估区域碳收支平衡及制定科学有效的土壤碳调控管理措施提供依据。     

Microbial immobilisation and turnover of C labelled substrates in two arable soils under field and laboratory conditions

Microbial biomass C immobilisation and turnover were studied under field and laboratory conditions in soils of high yield (HY) and low yield (LY) areas within an agricultural field. We compared the size and activity of soil microbial biomass (SMB) in the soils of the different yield areas under field and laboratory conditions. Soils were amended with ¹³C labelled mustard (Sinapis alba) residues (both experiments) and labelled glucose (laboratory only) at 500 μg C g⁻¹ dry soil. SMB-C, dissolved organic carbon (DOC) and total C content were monitored in the field and the laboratory. CO₂-efflux was also measured in laboratory treatments. Isotope ratios were determined for SMB in both experiments, but other variables only in the laboratory treatments. A positive priming effect was measured in three of four laboratory treatments. Priming was induced after a significant increase of soil derived C in the microbial biomass. Thereafter, the total C loss through priming was always smaller than or equal to the decline in microbial biomass C. In field and laboratory experiments SMB in the HY soil immobilised less of the added substrate C than LY soil SMB. Calculated turnover times in the laboratory glucose amendment were 0.24 (HY) and 0.31 y (LY), in the laboratory mustard treatment 0.58 (HY) and 0.44 y (LY) and in the field mustard amendments 1.09 (HY) and 1.25 y (LY). In both the field mustard and laboratory glucose treatments turnover in the HY soil tended to exceed that in the LY soil. These turnover times as well as the reaction of SMB-C to drying-rewetting and substrate addition, indicated that the HY soil possessed a more active microbial community with a more rapid C turnover than the LY soil. As C turnover is considered to be closely linked to nutrient cycles, faster turnover in the HY soil may involve a better nutrient supply for crops resulting in higher agricultural yield.     

Microbial immobilisation and turnover of N labelled substrates in two arable soils under field and laboratory conditions

Microbial biomass N dynamics were studied under field and laboratory conditions in soils of high yield (HY) and low yield (LY) areas in an agricultural field. The objective of the study was to determine the size and activity of soil microbial biomass in the soils of the different yield areas and to compare these data obtained under field and laboratory conditions. Soils were amended with ¹⁵N labelled mustard (Sinapis alba) residues (both experiments) and labelled nitrate (laboratory only) at 30 μg N g⁻¹ dry soil. Soil microbial biomass (SMB) N, mineral N (N_min) and total N content was monitored both in the field and in the laboratory. N₂O efflux was additionally measured in laboratory treatments. Isotope ratios were determined for SMB in both experiments, for all other parameters only in the laboratory treatments. In the laboratory less amounts of added substrate N were immobilised by the SMB in HY soils compared to LY soils, whereas in the field immobilisation of added N by SMB was higher in HY soils initially and slightly lower after 40 days of incubation. Calculated turnover times in the laboratory nitrate, laboratory mustard and field mustard amendments were 0.18, 0.27 and 0.74 years (HY) and 0.22, 0.61 and 1.01 years (LY), respectively. The turnover times of added substrate N always showed the trend to be faster in HY soils compared to LY soils. A faster turnover of nutrients in the HY soils may involve a better nutrient supply of the plants, which coincides with the higher agricultural yield observed in these areas.     

Photodegradation of rotenone in soils under environmental conditions

An environmental fate study was performed to analyze the effects of soil components on the photochemical behavior of rotenone. Photodegradation experiments were carried out on three types of soil collected in southern Italy, Valenzano (VAL), Turi (TUR), and Conversano (CON), from April to June 2006. Soil thin-layer plates (1 mm thick) were spiked with 1.5 mg/kg of rotenone and exposed under natural conditions of sunlight and temperature. The plates were removed from the sunlight at predetermined intervals of continuous irradiation. Other soil samples, control and sterilized, were kept in the dark to evaluate possible effects of chemical and microbiological degradation during the irradiation experiment. The time for 50% loss of the initially applied rotenone varied from 5 to 7 h, following the order TUR < CON < VAL. In environmental studies, changes in temperature and/or moisture affected the degradation rate and caused deviations from first-order kinetics. The photolysis reaction fit the two compartment or the multiple compartment model pathways better. A fast initial decrease during the first 5 h of rotenone irradiation was followed by a much slower decline, which clearly indicates the rather complex chemical process of rotenone photodegradation on soil surfaces. Also, the degradation was shown to be directly related to the soil concentration of clay and organic matter. Rotenolone (12abeta-hydroxyrotenone) was detected by HPLC/DAD/MS analysis as the only photodegradation byproduct of rotenone in soil thin layers. Results provide additional insights on the rates and the mechanisms of rotenone degradation, aiming to describe more clearly the degradation performance of chemical residues in the environment.     

Improved HPLC-MS/MS method for determination of isoxaflutole (balance) and its metabolites in soils and forage plants

A robust multi-residue procedure is needed for the analysis of the pro-herbicide isoxaflutole and its degradates in soil and plant materials at environmentally relevant (<1 microg kg-1) levels. An analytical method using turbo-spray and heat-nebulizer high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for the analysis of isoxaflutole (IXF) and its two metabolites, diketonitrile (DKN) and the benzoic acid metabolite (BA), at sub-microgram per kilogram levels in soil and plant samples. The average recoveries of the three compounds in spiked soil and plant samples ranged from 84 to 110% and 94 to 105%, respectively. The limits of quantification were validated at 0.06 microg kg-1 for soil and 0.3 microg kg-1 for plant samples. The limits of detection (LOD) for soil analysis were 0.01, 0.002, and 0.01 microg kg-1 for IXF, DKN, and BA, respectively. Corresponding LOD for the plant analysis method were 0.05, 0.01, and 0.05 microg kg-1. The developed method was validated using forage grass and soil samples collected from a field lysimeter study in which IXF was applied to each of four forage treatments. Forage plants and soils were sampled for analyses 25 days after IXF application to the soil. In soils, IXF was not detected in any treatment, and DKN was the predominant metabolite found. In forage plants, the concentrations of DKN and BA were 10-100-fold higher than that in soil samples, but IXF was not detected in any forage plants. The much higher proportion of BA to DKN in plant tissues (23-53%), as compared to soils (0-5%), suggested that these forages were capable of detoxifying DKN. The developed methods provided LODs at sub-microgram per kilogram levels to determine the fate of IXF and its metabolites in soils and forage plants, and they also represent considerable improvements in extraction recovery rates and detection sensitivity as compared to previous analytical methods for these compounds.