Effect of herbicides on glutathione S-transferases in the earthworm, <Emphasis Type="Italic">Eisenia fetida</Emphasis>

AIM AND BACKGROUND: Earthworms have been studied as a readily available, easily maintainable and cheap test species for assessing chemical pollution, and may be an alternative to in vivo rodent bioassays. The current investigation aims to characterize detoxification enzymes in Eisenia fetida and stress response against two herbicides with different modes of action, namely, fenoxaprop and metolachlor. METHODS: Herbicides were applied to soil containing earthworms. Animals were then collected, sacrificed and shock-frozen. Extracted protein was analyzed for glutathione S-transferase (GST) activity using CDNB (1-chloro-2,4-dinitrobenzene), DCNB (1,2-dichloro-4-nitrobenzene), pNBC (p-nitrobenzylchloride), PNOBC (p-nitrobenz-o-ylchloride) and selected herbicides. GST isoenzymes were partially purified by affinity chromatography and molecular weights were estimated by SDS-PAGE. RESULTS: In E. fetida protein extracts, GST activity towards model compounds ranked as CDNB>DCNB>PNBOC>PNBC. Fluorodifen was not conjugated at all, but fenoxaprop and metolachlor were conjugated at low rates. Furthermore, the GST isoenzyme pattern changed during the incubation with herbicides, either due to stress or as a defense reaction. After incubation with monochlorobimane, a strong fluorescence of the intestinal tract and the intersegments was observed, indicating organ-specific GST induction. DISCUSSION: According to the author's knowledge, here, for the first time, evidence is presented that E. fetida GST are also capable of conjugating a wider range of xenobiotic substrates. Different forms of GST were observed and changes in GST isoforms due to the herbicide treatment were also noticed. GST conjugation rates varied between different herbicides used in this experiment. It might be assumed that herbicides may well be detoxified by earthworms, to a certain extent, but that they are also potent stress factors influencing the detoxification system of the animal. High doses or long exposure might lead to deleterious effects on earthworms and limit their survival rate. The use of the animals as bioindicators for herbicides and herbicide residues seems very promising, but is surely influenced by the lack of detoxification for some compounds. CONCLUSIONS: Conjugation of several xenobiotics with model substances and herbicides is proven in the earthworm E. fetida. However, E. fetida has only limited capabilities of detoxifying herbicidal compounds. Different isoforms of GST were involved and altered in their activity after treatment. RECOMMENDATIONS AND PERSPECTIVES: The accumulation of GS-conjugates and their determination via fluorescence microscopy is a quick and secure, additional marker for exposure that should be further developed to complement existing biotests. The described methods and endpoints might help to understand the complex reaction of earthworms towards herbicides and lead to an adapted test methodology.     

Sonophotocatalytic degradation of alazine and gesaprim commercial herbicides in TiO2 slurry

The photocatalytic degradation of alazine and gesaprim commercial herbicides was carried out in aqueous TiO(2) suspensions under UV light (15W, 352 nm). Degradation of these herbicides was also observed by the combined effects of photocatalysis with sonolysis (sonophotocatalysis) using an ultrasound source of 20kHz. Degradation profiles were recorded by measuring the concentration of the active compounds present in the alazine (alachlor and atrazine) and gesaprim (atrazine) by HPLC as a function of irradiation time (sound and/or light). Over 90% of the active component in the gesaprim was abated and those in alazine were completely degraded. The content of total organic carbon and chemical oxygen demand was also monitored. Mineralization of the commercial herbicides was achieved. Over 80% of chemical oxygen demand abatement was attained for both herbicides with sonophotocatalysis at 150 min of irradiation time. The photocatalytic degradation of the herbicides followed a pseudo-first order kinetics and their rate constant was increased by the combined effects of sonolysis.     

Microbial phytotoxins as potential herbicides

Abstract

Microbes are sources of a diverse array of phytotoxic compounds. These compounds are generally structurally different from commercial herbicides, targeting different molecular sites of action within the plant. These novel structures and sites can be excellent leads for the discovery and development of safer synthetic herbicides. Microbial phytotoxins are often more environmentally benign than synthetic herbicides. Examples of phytotoxins from fungi (AAL‐toxin, cornexistin, cyperin, and tentoxin) with novel structures and sites of action are discussed. AAL‐toxin is toxic to a wide variety of weeds at very low dose rates. AAL‐toxin and many of its analogues kill plants by inhibiting a ceramide synthase‐like enzyme, causing rapid accumulation of free sphingoid bases that disrupt membranes. Cornexistin appears to be metabolically cnverted to an inhibitor of certain aspartate amino transferase isoenzymes. Its activity can be reversed by feeding aspartate and glutamate or with tricarboxylic acid cycle intermediates. Its activity is much like that of (aminooxy)acetate. Cyperin is a diphenylether phytotoxin that inhibits protoporphyrinogen oxidase, but does not kill plants by this mechanism. It appears to have other effects on porphyrin metabolism. Tentoxin is toxic by two mechanisms. It disrupts chloroplast development by inhibiting the processing of a nuclear‐coded plastid protein, and it also inhibits photophosphorylation by acting as an energy transfer inhibitor of coupling factor 1 ATPase. Other examples of phytotoxins from microbes with promise as herbicides will be mentioned.     

Air concentrations of currently used herbicides and legacy compounds in the Canadian prairies,subarctic, and arctic

Passive air samplers were installed in the summers of 2005 and 2007 for 90 days at four locations in the agricultural region of the Canadian Prairies and at five locations in the Canadian Subarctic and Arctic. The presence and masses of ten currently used herbicides and three legacy compounds in the polyurethane foam disks were quantified. Herbicides 2,4-D, bromoxynil and MCPA were detected at all locations in the Canadian Prairies and in both years because these herbicides are widely applied to control broadleaf weeds in cereal crops that are an integral part of Prairie agricultural production systems. MCPA was also detected at one location in the Arctic in 2007. The detection of the other seven herbicides in the 2 years combined ranged from no detections (atrazine only) to five detections for the relatively volatile herbicides trifluralin and triallate. Triallate was the only other herbicide detected in the Arctic (2005). Legacy compounds were either not detected (alachlor) or at levels near their detection level (γ-HCH and α-HCH). γ-HCH and α-HCH were more frequently detected in 2005 than in 2007 indicating that their concentrations in Canadian air have decreased over time. γ-HCH, widely used as an insecticide in Prairie oilseed production until 2002, was detected at larger concentrations in the Canadian Prairies than in the Subarctic and Arctic. α-HCH, a manufacturing by-product in technical HCH prior to 1971 in Canada, was not detected in the Canadian Prairies but was at detectable levels in the Subarctic and Arctic as the Arctic Ocean is reported to be a major source of α-HCH to the atmosphere. We conclude that some of the most widely used herbicides in Canadian agriculture today are commonly present in the air in regions where they are applied and that a portion of these herbicides may be traveling as parent molecules to the Canadian Arctic. To the authors’ knowledge, this is the first evidence of the presence of MCPA and triallate in Arctic air samples, perhaps because previous research has seldomly monitored for currently used herbicides in this region.     

A Comparison of Various Sources of Automotive Emissions

Chlorinated phenolic hydrocarbons are used as intermediate chemicals in the manufacture of 2,4-D (2,4-Dichlorophenoxy-acetic Acid) and related herbicides. These chemicals have an unusual odor with an extremely low threshold level of detection and consequently the manufacture and handling of these compounds poses a difficult odor control problem. Chipman Chemical Company at its Portland, Oregon plant has developed a system of fume collection and caustic soda solution scrubbing capable of removing phenolic compounds in the plant exhaust air to an acceptable level from an odor release standpoint. This article describes the development, present status, and projected future improvements of this odor control system.     

Effects of low glyphosate-based herbicide concentrations on endocrine-related gene expression in the decapoda Macrobrachium potiuna

Environmental Science and Pollution Research - Glyphosate-based herbicides (GBH) are the most used herbicides worldwide and are considered as endocrine-disrupting compounds (EDC) for non-target...     

Monitoring of triazine and phenylurea herbicides in the surface waters of Greece

A large-scale study was implemented to monitor triazine and phenylurea herbicides in the main surface water bodies of continental Greece from October 1998 to September 1999. Samples from 10 rivers and 7 lakes were analyzed for the presence of five triazine (atrazine, cyanazine, prometryne, simazine, terbuthylazine) and five phenylurea (chlorotoluron, diuron, linuron, metobromuron, monolinuron) herbicides. The samples were extracted with C18 cartridges and analyzed by high-performance liquid chromatography-diode array detection (HPLC-DAD). The most frequently detected herbicides were atrazine, followed by prometryne, cyanazine, and simazine. The concentrations of the compounds were generally low (< 0.78 micro g/L) and are not considered harmful for the freshwater ecosystem. Most of the positive samples were taken from the water bodies of northern Greece where agricultural activity is more intense.     

Herbicide safeners: uses,limitations, metabolism,and mechanisms of action

Several methods were examined to minimize crops injury caused by herbicides. Thus increase their selectivity. A selective herbicide is one that controls weeds at rates that do not injure the crop. Herbicides are selective in a particular crop within certain limits imposed by the herbicide, the plant, the application rate, the method and time of application, and environment conditions. Herbicide safeners are compounds of diverse chemical families. They are applied with herbicides to protect crops against their injury. Using chemical safeners offer practical, efficient and simple method of improving herbicide selectivity. This method has been applied successfully in cereal crops such as maize, rice and sorghum, against pre-emergence thiocarbamate and chloroacetanilide herbicides. Some reports indicate promising results for the development of safeners for post-emergence herbicides in broadleaved crops. Various hypotheses were proposed explaining mechanisms of action of herbicide safeners: interference with uptake and translocation of the herbicide, alteration in herbicide metabolism, and competition at site of action of the herbicide. Even though progress was made in the development of herbicide safeners and in understanding their mechanisms of action, more research is needed to elucidate clearly how these chemicals act and why their activity is restricted to particular crops and herbicides.     

Multivariate analysis of photo-Fenton degradation of the herbicides tebuthiuron, diuron and 2,4-D

The degradation of herbicides in aqueous solution by photo-Fenton process using ferrioxalate complex (FeOx) as source of Fe2+ was evaluated under blacklight irradiation. The commercial products of the herbicides tebuthiuron, diuron and 2,4-D were used. The multivariate analysis, more precisely, the response surface methodology was applied to evaluate the role of FeOx and hydrogen peroxide concentrations as variables in the degradation process, and in particular, to define the concentration ranges that result in the most efficient degradation of the herbicides. The degradation process was evaluated by the determination of the remaining total organic carbon content (TOC), by monitoring the decrease of the concentrations of the original compounds using HPLC and by the chloride ion release in the case of diuron and 2,4-D. Under optimized conditions, 20 min were sufficient to mineralize 93% of TOC from 2,4-D and 90% of diuron, including oxalate. Complete dechlorination of these compounds was achieved after 10 min reaction. It was found that the most recalcitrant herbicide is tebuthiuron, while diuron shows the highest degradability. However, under optimized conditions the initial concentration of tebuthiuron was reduced to less than 15%, while diuron and 2,4-D were reduced to around 2% after only 1 min reaction. Furthermore, it was observed that the ferrioxalate complex plays a more important role than H2O2 in the photodegradation of these herbicides in the ranges of concentrations investigated.     

Monitoring of Triazine and Phenylurea Herbicides in the Surface Waters of Greece

A large-scale study was implemented to monitor triazine and phenylurea herbicides in the main surface water bodies of continental Greece from October 1998 to September 1999. Samples from 10 rivers and 7 lakes were analyzed for the presence of five triazine (atrazine, cyanazine, prometryne, simazine, terbuthylazine) and five phenylurea (chlorotoluron, diuron, linuron, metobromuron, monolinuron) herbicides. The samples were extracted with C18 cartridges and analyzed by high-performance liquid chromatography–diode array detection (HPLC-DAD). The most frequently detected herbicides were atrazine, followed by prometryne, cyanazine, and simazine. The concentrations of the compounds were generally low (< 0.78 μ g/L) and are not considered harmful for the freshwater ecosystem. Most of the positive samples were taken from the water bodies of northern Greece where agricultural activity is more intense.     

Critical review of actually available chemical compounds for prevention and management of cyanobacterial blooms

Cyanobacteria proliferation is among the most threatening consequences of freshwater pollution. Health risks from human and other-organism exposure to cyanobacteria have led to an effort to find practical methods for cyanobacterial water-bloom reduction. Hence, methods and techniques have been developed in order to reduce the amount of phosphorus or to decrease the abundance of nuisance phytoplankton species directly in the water bodies (in-lake measures). Although these “acute” methods do not solve the problem of catchment area eutrophication, they are cheaper, easier to manage, and for some areas they are the only way to protect human and environmental health against massive cyanobacterial proliferation. This review summarizes the extent of knowledge and published data about the management using metals (Al, Fe, Cu, Ag, Ca), photosensitizers (hydrogen peroxide, phthalocyanines, TiO₂), herbicides and chemicals derived from natural compounds as fast and efficient removal agents of cyanobacteria. This review concludes that some compounds, when non-persistent and ecotoxicologically acceptable may help to manage cyanobacterial blooms in an efficient way compared to previous methods (e.g. copper sulfate).     

Dissipation of racemic mecoprop and dichlorprop and their pure R-enantiomers in three calcareous soils with and without peat addition

Two racemic herbicides, mecoprop (R,S-MCPP) and dichlorprop (R,S-DCPP), as well as their enantiopure R-forms, were incubated in three calcareous soils at 15 degrees C and 80% of their field capacity to try to elucidate their behaviour in soil and compare the dissipation rates when racemic and enantiopure compounds are used. Quantitation of pesticides is made by HPLC and the R/S ratio by GC-MS. The inactive S-enantiomer from the racemic forms persists longer than the R-forms in silt and sandy loam soils, but for shorter time in the clay loam soil. The pure R-enantiomers, both for MCPP and DCPP, after incubation in soil, are partially converted into their S-forms. In all cases, the dissipation of racemic and pure enatiomeric forms is lower in the clay loam soil than in the silt and sandy loam soils. The R-forms' peristence, in the three soils, is approximately two times lower when they are incubated alone than when they are incubated as racemic compounds. When peat is added, the persistence of these herbicides in the silt and sandy loam soils increases, while in the clay loam soil it decreases. Besides, in the clay loam soil, the enantiomeric ratio (ER) changes from its S-preferential degradation to a preferential degradation of its R-form, so an increase in the persistence of the inactive S-form occurs.     

Multiple resistance of acetolactate synthase and protoporphyrinogen oxidase inhibitors in Euphorbia heterophylla biotypes

Resistance to acetolactate synthase (ALS)-inhibiting herbicides in Brazil has been documented for six species. The probability to select biotypes of Euphorbia heterophylla (EPPHL) with multiple resistance increases in the same order of magnitude as the use of other herbicides belonging to only one mechanism of action. The objectives of this work were to evaluate the distribution of resistant populations (R) in the states of the Parana and Santa Catarina; to determine the existence of populations of EPHHL with multiple resistance to ALS and PROTOX inhibitors, and to confirm the occurrence of cross resistance to compounds of these mechanisms of action. Seeds of EPHHL of areas with suspected resistance had been sampled in 97 places during 2003. In the greenhouse experiment samples of each population were sprayed with imazethapyr or fomesafen, at only one rate. To identify the resistant ones they were sprayed with different levels of the herbicides imazethapyr and fomesafen. Later they were sprayed with diverse herbicides of the same mechanisms of action to confirm the multiple/cross resistance. There is widespread distribution in the region of populations with resistance to ALS inhibitors. Some biotypes demonstrated resistance to herbicides from the two mechanisms of action. The resistance factor (FR), or the relation of resistance between R and susceptible biotypes, confirms the existence of two biotypes of EPHHL with cross resistance to several herbicides inhibitors of ALS and PROTOX.     

Release behavior of triazine residues in stabilised contaminated soils

This paper reports the release behavior of two triazines (atrazine and simazine) in stabilised soils from a pesticide-contaminated site in South Australia. The soils were contaminated with a range of pesticides, especially with triazine herbicides. With multiple extractions of each soil sample with deionised water (eight in total), 15% of atrazine and 4% of simazine residues were recovered, resulting in very high concentrations of the two herbicides in leachate. The presence of small fractions of surfactants was found to further enhance the release of the residues. Methanol content up to 10% did not substantially influence the concentration of simazine and atrazine released. The study demonstrated that while the stabilisation of contaminated soil with particulate activated carbon (5%) and cement mix (15%) was effective in locking the residues of some pesticides, it failed to immobilise triazine herbicides residues completely. Given the higher water solubility of these herbicides than other compounds more effective strategies to immobilise their residues is needed.     

Progress in herbicide determination with the thylakoid bioassay

Chloroplast thylakoids are used as biological units to determine herbicides in different kinds of water samples as well as in aqueous extracts of compost, soil or food samples. The thylakoid bioassy shows clearly inhibition of fluorescence yield in the presence of photosystem II specific herbicides. Due to this method the ecotoxicological effect of samples with unknown pollutants can be tested fast and cost effective. It has been proven that all photosynthetic active compounds are recorded at the same time because only additive interactions occur. Therefore, the contamination level can be expressed as cumulative parameter for photosystem II active substances. Application was improved clearly by the addition of the radical scavenger sodium ascorbate to the isolation media and by a higher concentration of the measuring medium. A new data evaluation method is described yielding in a lower detection limit of 0.4 μg diuron/1. The guidelines for the quality of water for human consumption with an allowable concentration of pesticides in groups is 0,5 μg/1 [1,2] and can be controlled with the thylakoid bioassay without performing any preconcentration steps.     

Validation of the species sensitivity distribution in retrospective risk assessment of herbicides at the river basin scale—the Scheldt river basin case study

Species sensitivity distribution (SSD) is commonly used in prospective risk assessment to derive predicted no-effect concentrations, toxicity exposure ratios, and environmental quality standards for individual chemicals such as pesticides. The application of SSD in the retrospective risk assessment of chemical mixtures at the river basin scale (i.e., the estimation of “multiple substance potentially affected fractions” [msPAFs]) has been suggested, but detailed critical assessment of such an application is missing. The present study investigated the impact of different data validation approaches in a retrospective model case study focused on seven herbicides monitored at the Scheldt river basin (Belgium) between 1998 and 2009. The study demonstrated the successful application of the SSD approach. Relatively high impacts of herbicides on aquatic primary producers were predicted. Often, up to 40 % of the primary producer communities were affected, as predicted by chronic msPAF, and in some cases, the predicted impacts were even more pronounced. The risks posed by the studied herbicides decreased during the 1998–2009 period, along with decreasing concentrations of highly toxic pesticides such as simazine or isoproturon. Various data validation approaches (the removal of duplicate values and outliers, the testing of different exposure durations and purities of studied herbicides, etc.) substantially affected SSD at the level of individual studied compounds. However, the time-consuming validation procedures had only a minor impact on the outcomes of the retrospective risk assessment of herbicide mixtures at the river basin scale. Selection of the appropriate taxonomic group for SSD calculation and selection of the species-specific endpoint (i.e., the most sensitive or average value per species) were the most critical steps affecting the final risk values predicted. The present validation study provides a methodological basis for the practical use of SSD in the retrospective risk assessment of chemical mixtures.     

Laboratory study on the interaction between herbicides and sediments in water systems

Interaction between herbicides and sediments in water systems is an important process occurring in water, which influences the behaviour of the herbicides in water. This paper reports on the sorption of herbicides norflurazon, oxadiazon and trifluralin on soil and the interaction between the herbicides and sediments under stirred and non-stirred conditions. The sorption coefficients of the herbicides on soils are 3.58 and 5.41 for norflurazon, 23.43 and 28.07 for oxadiazon and 890.73 and 1217.20 for trifluralin. The sorption of the herbicides is related to the organic carbon content in the soils. This study shows a greater sorption of the herbicides on stirred sediments than on non-stirred sediments due to more significant contact under stirred conditions. The relative concentrations of the herbicides in water systems containing sediments were higher than those in pure water 6 and 13 days after treatment. When these herbicides were sorbed on sediments, their persistence in water increased. Sorption of herbicides on sediments in aquatic systems could protect them from degradation in water.     

Herbicide contamination of surficial groundwater in Northern Italy

Data on herbicide pollution in groundwater are rather scarce; monitoring data are based on single investigation, focussing on limited area and on few compounds of interest. The large number of approved active ingredients (approximately 600 chemicals) makes difficult to obtain an accurate and actual information on herbicide application in different countries, even if herbicides are the second most important class of pesticides used in the European Union. The results of a two-year monitoring campaign undertaken in two areas intensively cultivated at Lombardy, Northern Italy, showed a diffuse groundwater contamination due to active ingredients and their metabolites. More than 50% of samples overcame M.A.C. and the most common herbicides were Atrazine, Terbuthylazine and Metolachlor, while DEA and DET metabolites were often characterized by greater concentrations than their relative active principles.     

Simultaneous analysis of triasulfuron, metsulfuron-methyl and chlorsulfuron in water and alkaline soils by high-performance liquid chromatography

A method capable of simultaneously detecting residues of three sulfonylurea herbicides at microgram/l and microgram/kg level in water and alkaline soils has been described. The method is based on solid phase extraction and HPLC with UV detection. In alkaline soils especially those containing low organic carbon it was possible to extract the herbicides with de-ionised water and no clean up step was needed. Soil samples spiked with technical grade triasulfuron, metsulfuron-methyl and chlorsulfuron were extracted twice by shaking with de-ionised water for one hour and centrifuging at 10,000 rpm for 15 minutes. Supernatants filtered through glass micro-fibre filters were passed through C18 cartridges previously pre-conditioned with methanol and de-ionised water at a flow rate of < 20 ml/min. Residues of the herbicides retained on the cartridge were eluted with acidified methanol. The eluate was analysed by HPLC. A C18 column was used with a mobile phase of methanol/water (40 + 60, V/V for for the herbicide residues were 1.0 microgram/l and 3 micrograms/kg in water and soil, respectively. The average recoveries for water samples ranged from 73-94%, while for soil samples recoveries were 77-97% for the three compounds studied.     

Sorption and desorption kinetics of diuron, fluometuron, prometryn and pyrithiobac sodium in soils

The sorption and desorption characteristics of four herbicides (diuron, fluometuron, prometryn and pyrithiobac-sodium) in three different cotton growing soils of Australia was investigated. Kinetics and equilibrium sorption and desorption isotherms were determined using the batch equilibrium technique. Sorption was rapid (> 80% in 2 h) and sorption equilibrium was achieved within a short period of time (ca 4 h) for all herbicides. Sorption isotherms of the four herbicides were described by Freundlich equation with an r2 value > 0.98. The herbicide sorption as measured by the distribution coefficient (Kd) values ranged from 3.24 to 5.71 L/kg for diuron, 0.44 to 1.13 L/kg for fluometuron, 1.78 to 6.04 L/kg for prometryn and 0.22 to 0.59 L/kg for pyrithiobac-sodium. Sorption of herbicides was higher in the Moree soil than in Narrabri and Wee Waa soils. When the Kd values were normalised to organic carbon content of the soils (Koc), it suggested that the affinity of the herbicides to the organic carbon increased in the order: pyrithiobac-sodium < fluometuron < prometryn < or = diuron. The desorption isotherms were also adequately described by the Freundlich equation. For desorption, all herbicides exhibited hysteresis and the hysteresis was stronger for highly sorbed herbicides (diuron and prometryn) than the weakly sorbed herbicides (fluometuron and pyrithiobac-sodium). Hysteresis was also quantified as the percentage of sorbed herbicides which is not released during the desorption step (omega = [nad/nde - 1] x 100). Soil type and initial concentration had significant effect on omega. The effect of sorption and desorption properties of these four herbicides on the off-site transport to contaminate surface and groundwater are also discussed in this paper.