Interaction of the 2,4-dichlorophenoxyacetic acid herbicide with soil organic matter moieties: a theoretical study

The determination of the structure of humic substances from soils and natural waters is an intriguing problem in soil science. Humic substances consist of molecules covering a broad distribution of molecular size and involving different functional groups. Taking this into account, we have chosen smaller model systems with functional groups typically present in humic substances. We investigated theoretically, by quantum chemical calculations, the environmental effects on the complexes formed from the interaction of 2,4‐dichlorophenoxyacetic acid and its anion with acetaldehyde, methanol, methylamine, protonated methylamine, acetic acid and water. The important case of a cation bridge mechanism, with Ca²⁺ as the bridging cation, is also included into the set of model reactions. It is found that this cation bridge belongs to one of the most stable mechanisms of fixation of organic compounds in soils. According to our calculations the hydroxyl group forms the most stable complexes with 2,4‐D in a polar solvent environment.     

Two enzyme immunoassays to screen for 2,4-dichlorophenoxyacetic acid in water

Two solid-phase enzyme immunoassays were developed to measure 2,4-dichlorophenoxyacetic acid (2,4-D), using 2 sets of structurally distinct immunogens and enzyme ligands. The 2,4-D analog, 2-methyl-4-chlorophenoxyacetic acid (MCPA), gave a similar response with both methods, whereas other phenoxy herbicides cross-reacted differently. In method A, the aromatic moiety of 2,4-D was distal from the carrier protein and labeled enzyme, whereas in method B, the acetic acid portion of the herbicide was distal. The use of both methods to screen for this herbicide in ground water and municipal and river water reduced the number of false-positive responses. Water sources having a low background response could be monitored with either method alone. When a concentration step, with disposable C18 extraction columns, was used, the limit of sensitivity was 5 micrograms/L. Method A was the more sensitive of the 2 methods with a limit of detection of 10 micrograms/L without the concentration step.     

Metabolism of 2,4-dichlorophenoxyacetic acid in laying hens and lactating goats

2,4-Dichlorophenoxyacetic acid (2,4-D) labeled with (14)C was found to be rapidly eliminated by laying hens and lactating goats dosed orally for 7 consecutive days at 18 mg/kg of food intake and for 3 consecutive days at 483 mg/kg of food intake, respectively. Excreta of hens and goats contained >90% of the total dose within 24 h after the final dose. Tissue residues were low and accounted for <0.1% of the dose in these animals. For hens, the residues in muscle, liver, and eggs (0.006-0.030 ppm) were lower than those found in fat and kidney (0.028-0.714 ppm), 2,4-D equivalents. The tissue with highest residue in goat was the kidney at 1.44 ppm, 2,4-D equivalents. Milk, liver, composite fat, and composite muscle had significantly lower residue levels of 0.202, 0.224, 0.088, and 0.037 ppm, respectively. The most abundant tissue residue was 2,4-D and acid/base releasable residues of 2,4-D. A minor metabolite was identified as 2,4-dichlorophenol.     

Response of soil microbial communities to the herbicide mesotrione: A dose-effect microcosm approach

Mesotrione is a new selective herbicide used for maize crops. The responses of microbial communities of a chernozem soil (Limagne basin, France) to pure or formulated (Callisto^®) mesotrione, applied at three different doses [one fold field rate (1 × FR), 10 × FR and 100 × FR], were studied using a laboratory microcosm approach. The effects were assessed on the prokaryotic cell abundance, the overall microbial activities (substrate-induced respiration (SIR) and dehydrogenase activity (DHA)) and the genetic structure of the bacterial and fungal communities (temporal temperature/denaturing gradient gel electrophoresis (TT/DGGE)). Mesotrione dissipation was similar whatever the formulation applied and the amounts dissipated were positively correlated to application rates. Several biodegradation products including the metabolites 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) and 2-amino-4-methylsulfonylbenzoic acid (AMBA) were detected from day 42 post-treatment, in 10 × FR and 100 × FR treated soils. No response of the soil microbial communities was detected in soil spread with both the 1 × FR applications. Overall soil microbial activity was stimulated from day 6 by 10 × FR of Callisto^® and more strongly by 100 × FR of pure mesotrione and Callisto^®, whereas prokaryote abundance did not increase before day 95 in both the 100 × FR treatments. Genetic structural shifts recorded from day 42 in the bacterial and fungal communities were small and mainly attributable to variations in band intensity. Maximum dissimilarity of the bacterial and fungal genetic structures between control and 100 × FR treated soils did not exceed 12% and 28%, respectively. The general pattern was that more consistent effects occurred with increasing exposure times, especially in both the 100 × FR treated soils. These microbial responses could be due to the stimulation of (i) adapted mesotrione-degrading microorganisms and (ii) the activity of resistant heterotrophic microbial groups promoted by dead biomass from sensitive organisms. In addition, at 100 × FR doses, pure mesotrione seemed to induce stronger microbial responses than Callisto^®, formulation which contains adjuvants with potential side-effects on some microbial populations. This experimental approach indicated that pure mesotrione and Callisto^® affected soil microbial communities, but the effects were only detected at doses far exceeding the recommended field rates.     

Soil organic carbon and microbial communities respond to vineyard management

The farming practices in vineyards vary widely, but how does this affect vineyard soils? The main objective of this study was to evaluate the effects of vineyard management practices on soil organic matter and the soil microbial community. To this end, we investigated three adjacent vineyards in the Traisen valley, Austria, of which the soils had developed on the same parent material and under identical environmental/site conditions but were managed differently (esp. tillage, fertilizer application, cover crops) for more than 10 yrs. We found that topsoil bulk density (BD) decreased with increasing tillage intensity, while subsoil BD showed the opposite trend. Soil organic carbon (SOC) stocks in 0–50 cm depth increased from 10 kg m^?2 in an unfertilized and frequently tilled vineyard to 17 kg m^?2 in a regularly fertilized but less intensively tilled vineyard. Topsoil microbial biomass per unit SOC, estimated by the sum of microbial phospholipid fatty acids (PLFAs), followed this trend, albeit not statistically significantly. Principal component analysis of PLFA patterns revealed that the microbial communities were compositionally distinct between different management practices. The fungal PLFA marker 18:2ω6,9 was highest in the vineyard with the lowest amount of extractable Cu (by 0.01 m CaCl₂), and the bacterial‐to‐fungal biomass ratio was positively correlated with extractable Cu. Our results indicate that tillage and fertilizer application of vineyards can strongly affect vineyard soil properties such as BD and SOC stocks and that the application of Cu‐based fungicides may impair soil fungal communities.     

Fluorescent caged compounds of 2,4-dichlorophenoxyacetic acid (2,4-d): photorelease technology for controlled release of 2,4-D

A novel controlled-release formulation (CRF) of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was developed to reduce its negative environmental impacts by improving its herbicidal efficacy. The 2,4-D was chemically caged by coupling with photoremovable protecting groups (PRPGs) of coumarin derivatives. Photophysical studies of caged compounds showed that they all exhibited strong fluorescence properties. Controlled release of 2,4-D was achieved by irradiating the caged compounds using UV-vis light (310, 350, and 410 nm). The effect of various factors such as pH, solvent, and different substituents at the seventh position of coumarin moiety on the rate of photorelease was studied. The herbicidal activity of caged compounds and 4-(hydroxymethyl)-7-substituted coumarins was studied against Vigna radiata . The new formulation provided greater control over the release of 2,4-D by UV-vis light and also demonstrated the potential of the PRPGs not only to act as a delivery device but also to possess herbicidal activity after photorelease.     

DNOC, a model pollutant, adversely affects the potential of soil microbial communities to mineralise the herbicide 2,4-D: an investigation using micro-sampling procedures

The effects of the herbicide, DNOC, 4,6-dinitro-o-cresol, a model pollutant, have been studied by comparing the potential of soil microbial communities present in individual soil aggregates or in larger soil microcosms as samples of soil aggregates to mineralise the herbicide 2,4-D. We have shown that 2-3 mm soil aggregates vary widely in their 2,4-D mineralisation potential and that ageing or exposure to DNOC considerably simplified the distribution patterns of this capacity. The main factors of variation have been quantified and classified using a quasi-likelihood method derived from the Generalised Linear Model approach. Besides DNOC concentration and duration of exposure, an additional ‘rank’ factor reflecting a desiccation gradient of the aggregates on the microtiter plates was found to have statistical significance. We concluded that it should be possible to derive an experimental approach, designated as ‘functional profiling’, with potential use to detect soil chemical contamination. Curves of 2,4-D mineralisation in individual soil aggregates could be classified according to three different types of kinetics, which were assumed to reflect heterogeneous spatial distribution, differences in microbial community composition and varying efficiency of the microbial consortia involved in 2,4-D degradation. Exposure to DNOC considerably simplified the distribution patterns of the different types of kinetics with one type, showing slow rate and low cumulative mineralisation, becoming predominant as ageing, concentration and duration of DNOC exposure increased. We argue on the possible use of ‘kinetic profiling’ as a sensitive bioindicator of soil quality. By comparison, in soil microcosms, 2,4-D mineralisation showed an extra mineralisation potential of 64% over individual aggregates in the control soil and exposure to DNOC was followed by concentration and time-dependent recovery of the 2,4-D mineralisation potential. It is likely that 2 g size soil microcosms gather a larger number of biochemical capacities which could complement each other to increase the potential of soil to mineralise xenobiotic compounds.     

Soil microbial communities resistant to changes in plant functional group composition

The soil community is an often ignored part of research which links plant biodiversity and ecosystem functioning despite their influence on numerous functions such as decomposition and nutrient cycling. Few consistent patterns have been detected that link plant and soil community composition. We used a removal experiment in a northern Canadian grassland to examine the effects of plant functional group identity on soil microbial community structure and function. Plant functional groups (graminoids, legumes and forbs) were removed independently from plots for five growing seasons (2003-2007) and in the fifth year effects on the soil microbial community were examined using substrate-induced respiration (SIR - a measure of metabolic diversity) and phospholipid fatty acid analysis (PLFA - a measure of microbial community composition). Removal treatments were also crossed with both a fertilizer treatment and a fungicide treatment to determine if effects of functional group identity on the soil community were context dependent. Plant functional group identity had almost no effect on the soil microbial community as measured by either SIR or PLFA. Likewise, soil properties including total carbon, pH, moisture and nutrients showed a limited response to plant removals in the fifth year after removals. We found a direct effect of fertilizer on the soil community, with fertilized plots having decreased metabolic diversity, with a decreased ability to metabolize amino acids and a phenolic acid, but there was no direct soil microbial response to fungicide. We show that in this northern Canadian grassland the soil microbial community is relatively insensitive to changes in plant functional group composition, and suggest that in northern ecosystems, where plant material is only slowly incorporated into the soil, five growing seasons may be insufficient to detect the impact of a changing plant community on the soil microbes.     

Spatial variability of 2,4-dichlorophenoxyacetic acid (2,4-D) mineralisation potential at a millimetre scale in soil

We analysed the ability of soil units of millimetre size to mineralise a herbicide, 2,4-D, using incubations of individual aggregates (2-7 mm diameter) and 6×6×6 mm³ cubes dissected from soil cores, under standard conditions. Mineralisation of ¹⁴C-ring labelled 2,4-D was measured using a barite paper trap and a Phosphorimager to record the evolved ¹⁴C-CO₂ from these very small soil samples. We found a large variability of 2,4-D mineralisation potential between aggregate size classes, between individual aggregates of the same size and between the different dissected cubes from a given core. We explained this variability by an uneven distribution of the degrading microorganisms at this scale, and to a lesser extent, an uneven distribution of C, necessary for co-metabolism. Furthermore, we found that in a soil core, the dissected cubes with a large mineralisation potential were not randomly distributed, but rather organised into centimetre sized hot spots.     

Kinetic modeling of 2,4-dichlorophenoxyacetic acid (2,4-D) degradation in soil slurry by anodic fenton treatment

Anodic Fenton treatment (AFT) has been shown to be a promising technology in pesticide wastewater treatment. However, no research has been conducted on the AFT application to contaminated soils. In this study, the 2,4-D degradation kinetics of AFT in a silt loam soil slurry were investigated for the first time, and the effects of various experimental conditions including initial 2,4-D concentration, Fenton reagent delivery rate, amount of humic acid (HA) addition, and pH were examined. The 2,4-D degradation in soil slurry by AFT was found to follow a two-stage kinetic model. During the early stage of AFT (the first 4-5 min), the 2,4-D concentration profile followed a pseudo-first-order kinetic model. In the later stage (typically after 5 or 6 min), the AFT kinetic model provided a better fit. This result is most likely due to the existence of (*)OH scavengers and 2,4-D sorption on soil. The Fe(2+) delivery rate was shown to be a more significant factor in degradation rate than the H(2)O(2) delivery rate when the Fe(2+)/H(2)O(2) ratios were in the range of 1:2 to 1:10. The presence of HA in soil lowered the AFT rate, most probably due to the competition with 2,4-D for consumption of (*)OH and increased sorption of 2,4-D on soil. The optimal pH for 2,4-D degradation in soil slurry by AFT was observed to be in the range of pH 2-3.     

Microbial mineralization of atrazine and 2,4-dichlorophenoxyacetic acid in riparian pasture and forest soils

Microbial biomass and mineralization of atrazine [2-chloro-4(ethylamino)-6(isopropylamino)s-triazine] and 2,4-D (2,4-dichlorphenoxyacetic acid) were examined in the top 10 cm of riparian pasture soils and in the litter layer and top 10 cm of mineral soils of riparian forest ecosystems. The riparian forest litter had higher levels of active and total fungal biomass than forest or pasture mineral soils in winter, spring, and fall. Active bacterial biomass was higher in forest litter than in forest and pasture mineral soils in spring and autumn, and higher in forest mineral soils than in pasture soils in summer. Total bacterial biomass was higher in forest mineral soils than in pasture soils during all seasons. In spring, it was also higher in forest litter than in pasture soils. Atrazie and 2,4-D mineralization in pasture soils was exceeded by that in forest litter in spring and autumn and by that in forest mineral soils in summer and autumn. There was no correlation between either active or total fungal and bacterial biomass with pesticide degradation.     

The influence of nitrogen on atrazine and 2,4-dichlorophenoxyacetic acid mineralization in grassland soils

The influence of fertilizer N on the mineralization of atrazine [2-chloro-4(ethylamino)-6(isopropylamino)-s-triazine] and 2,4-D (2,4-dichlorophenoxyacetic acid) in soils was assessed in microcosms using radiometric techniques. N equivalent to 0, 250, and 500 kg N as NH₄NO₃ ha^-1 was added to three grassland soils. Compared to the control, the 250- and 500-kg treatments suppressed mineralization of atrazine by 75 and 54%, respectively, and inhibited mineralization of 2,4-D by 89 and 30%, respectively. Active fungal biomass responded to the N treatments in an opposite manner to herbicide mineralization. Compared to the control, the 250- and 500-kg treatments increased the active fungal biomass by more than 300 and 30%, respectively. These results agree with other observations that N can suppress the decomposition of resistant compounds but stimulate the primary growth of fungi. The degree of suppression was not related to the amount of N added nor to the inherent soil N levels before treatment. The interaction between the N additions and the active fungal biomass in affecting herbicide mineralization suggests that N may alter microbial processes and their use of C sources and thus influence rates of herbicide degradation in the field.