Herbicides

Herbicides are commonly used by farmers to get rid of deleterious weeds. There is a common believe by beekeepers that herbicides have no harmful impacts on bee colonies. In this study, the effects of tow
glyphosate herbicides (herbazed and glypho-up) on some parameters of honey bee workers were investigated. Three concentrations of each herbicide were tested. Concerning survival, the highest concentrations of
herbazed and glypho-up showed significant impacts on caged bees than the control group (bees supplied with sugar syrup only) while the low concentrations showed the vice versa. The highest concentration of glypho-up caused the highest reduction in survival of caged bees. The tested herbicides showed no significant impacts on the navigation ability of bee workers than the control group. The proboscis extension reflex test was used to evaluate the learning ability of bee workers. The results showed insignificant impacts of the tested herbicides on the learning ability of bee workers than the control group. The survival was low when Nosema infected bees were exposed to the high concentration of glypho-up unlike Nosema infected bees and healthy ones. This study has a significant contribution towards understanding the potential impacts of herbicides on honey bees.

Atrazine, a chlorotriazine herbicide, is used to control annual grasses and broadleaf weeds. In this review, we summarize our laboratory's work evaluating the neuroendocrine toxicity of atrazine (and related chlorotriazines) from an historic perspective. We provide the rationale for our work as we have endeavored to determine: 1) the underlying reproductive changes leading to the development of mammary gland tumors in the atrazine-exposed female rat; 2) the cascade of physiological events that are responsible for these changes (i.e., the mode of action for mammary tumors); 3) the potential cellular mechanisms involving adverse effects of atrazine; and 4) the range of reproductive alterations associated with this pesticide.

We studied the activity of 14 compounds, all of which have been shown to interfere in plant cell division, in two animal tumor cell cultures, EL-4 and L1210. Four compounds [propham, chlorpropham, bensulide S-(O,O-diisopropylphosphorodithioate) ester of A/-(2-mercaptoethyl)benzenesulfonamide), and siduron] had a 50% inhibitory dose less than 10~4 M; six [2,3,5-triiodobenzoic acid, (2,4-dichlorophenoxy)acetic acid, bromacil, (2,4,5-trichlorophenoxy)acetic acid, naptalam, and (4-chloro-2-methylphenoxy)acetic acid] had a 50% inhibitory dose between 10~" and 10~3 M, and the remaining four 2,3: 4,6-di-O-isopropylidene-2-keto-L-gulonate, eptam, maleic hydrazide, and 4-(methylsulfonyl)-2,6-dinitro-/\/,N,-dipropylaniline] had a 50% inhibitory dose at higher than 10~3 M. There was a significant correlation between the effect on the two cell lines as well as between the inhibition of cell proliferation and that of thymidine and leucine uptake. More detailed study of cell proliferation and leucine and thymidine uptake for bensu lide and 2,3,5-triiodobenzoic acid revealed a dose-response pattern of inhibition starting shortly after exposure of the cells to the compounds. These results indicate that some inhibitors of plant cell division are capable of inhibiting the proliferation of animal tumor cells.

Fluorochloridone (FLC) is a herbicide used worldwide that is thought to be safe. However, due to its potential genotoxicity, cytotoxicity, and even systematic toxicity, there are increasing concerns about human exposure to this compound. Thus, the metabolism and bioactivation of FLC was investigated. After oral administration to mice, 27 metabolites were identified by ultrahigh performance liquid chromatography-electrospray ionization-quadrupole time-of-flight-mass spectrometry and with further structural identification by nuclear magnetic resonance spectroscopy. Hydroxylation and oxidative dechlorination were the major phase I pathways, while glutathione (GSH) and N-acetylcysteine conjugations were two major phase II pathways, indicating the formation of a reactive intermediate. In vitro microsomal and cytosolic studies revealed that a GSH conjugate (M13) was the predominant metabolite of FLC formed through a nucleophilic SN2 substitution of 3-Cl by GSH; this pathway is NADPH indep...

Fractions of the acetone extract and a mixture of two diterpenes from the marine brown alga Dictyota menstrualis were prepared with the aim of identifying potential effects on the germination of seeds and on elongation of the radicle and hypocotyl of the weeds Mimosa pudica and Senna obtusifolia. The bioassay on seed germination was performed in controlled conditions of 25 o C temperature and a 12 hour photoperiod, while the one concerning radicle and hypocotyl elongation was performed at the same temperature, though adopting a photoperiod of 24 hours. The results varied according to the receptor species, the fraction utilized, and its concentration. TLC analysis of the fractions and comparison with isolated products indicated that the diterpenes pachydictyol A and isopachydictyol A were the most abundant compounds in fraction HE, whereas the diterpene 6-hydroxy-dichotomano-2, 13-diene-16, 17-dial (3) was the most abundant compound in fractions DC and EA. Analysis of less polar fractions (in n-hexane, dichloromethane and ethyl acetate) revealed values of less than 30% inhibition. On the other hand, the ethanol/water fraction was the most active in all instances. The biological activity of these fractions must be due to the presence of known diterpenes and/or sulfated polysaccharides isolated in earlier studies.

The effects of paraquat on rat brain were studied. Activities of complex I (NADH: ubiquinone oxidoreductase) in mitochondrial electron transport system, lipid peroxidation and the amount of catecholamines in rat brain were measured after acute paraquat exposure. Complex I activities were significantly lower and lipid peroxides were higher in the brains of a paraquat-treated group than in those of a control group. Lipid peroxide in rat serum, however, did not increase after paraquat exposure. A study of the time dependency of paraquat effects disclosed that mitochondrial complex I activities in rat brain as well as those in rat lung and liver gradually decreased prior to the appearance of respiratory dysfunction. As compared to controls, the dopamine in rat striatum was significantly lower in the paraquat-treated group. These results suggest that paraquat after crossing the blood-brain barrier might be reduced to the radical in rat brain, which may damage the brain tissue, especially dopaminergic neurons in striatum. We therefore propose that cerebral damage should be taken into consideration on paraquat exposure. Patients may therefore need to be followed up after exposure to high doses of paraquat.

In ecotoxicological studies involving community-level investigations, rapid and multiparametric fluorescence-based methods may provide substantial advantages over traditional methods used for structural and functional community analysis. Therefore, multiwavelength-excitation pulse-amplitude modulated (PAM) fluorometry was applied in this study to assess long-term changes in periphyton community structure, shortterm effects on periphyton functioning (photosynthesis) and pollution induced community tolerance (PICT). For inter-calibration, periphyton structure was evaluated by chemotaxonomic analysis of accessory pigments and a four-wavelength-excitation PAM fluorometer. Short-term effects of herbicides were evaluated by fluorescence quenching analysis and 14 C-incorporation as a proxy of primary production. Subsequently, the method was applied to assess structural and functional changes in periphyton communities after isoproturon exposure for 14 and 26 days, respectively. Results showed good correlation of the PAM fluorescence-based measurements with traditional methods for biofilms in the initial colonisation phase for structural and functional parameters. However, for biofilms older than 9 weeks PAM fluorescence may underestimate biomass. Multiwavelength-excitation PAM fluorometry showed good correlation with marker pigment concentrations indicating that this method provides a reliable estimate of the community structure. PAM fluorometry was able to quantify changes of biomass and follow relative shifts in class composition of biofilms under exposure of isoproturon. Short-term tests based on the quantification of the inhibition of the effective quantum yield revealed a concentration-dependent increase of PICT. The observation of two succession phases of the biofilms after 14 and 26 days of growth, respectively, revealed that sensitivity of biofilms decreased with increasing age and biomass, respectively, but PICT remained a characteristic parameter of exposed communities in a concentration-dependent relationship. In conclusion, multiwavelength-excitation PAM fluorometry has considerable potentials in multispecies and multiparameter assessment of toxic effects on community level in terms of (1) a combined and rapid evaluation of structural and functional parameters in parallel, (2) screening of trends over time, (3) observing effects in replication and (4) being non-destructive. The approach therefore provides a perspective for a better understanding of community-level effects as species interactions in terms of PICT and therefore a higher ecological realism in risk assessment of toxicants.

A solid-phase microextraction (SPME)-GC procedure has been developed for the analysis of four selected pesticides (propanil, acetochlor, myclobutanil and fenoxycarb) in water samples. Mass spectrometry (MS) was used and two different instruments, a quadrupole MS system and an ion trap operating in the MS-MS mode, were compared. A Carbowaxdivinylbenzene SPME fiber was used. The performances of the two GC-MS instruments were comparable in terms of linearity (in the range of 0.1-10 mg / l in water samples) and sensitivity (limits of detection were in the low ng / l range); the quadrupole MS instrument gave better precision than the ion trap MS-MS system, but generally the relative standard deviations for replicates were acceptable for both instruments (,15%). Specificity with these two instruments was comparable in the analysis of ground water samples. Recovery tests were made to assess the applicability of the SPME procedure in the quantitative analysis of contaminated groundwaters.

This report describes separation and detection of chlorophenoxy acid herbicides spiked in drinking water by the technique combining solid-phase extraction, fieldamplified sample stacking, capillary electrophoresis, and potential gradient detection. The herbicide solution (400 mL) was concentrated to 0.1 mL by the solid-phase extraction procedure. The buffer containing 3 mM ammonia and 0.3 mM hydroxypropyl-bcyclodextrin was adjusted to pH 9.0 with ammonia. The sample solution was injected into the capillary to 30% of the whole length, and 29 kV and 9 kV were employed for field-amplified sample stacking and separation, respectively. The herbicides were baseline separated and the detection limits with the above combined techniques were in the range of 1-4610 22 ng/mL.

Functional identity and significant similarities in cofactors and sequence exist between the L and M reaction center proteins of the photosynthetic bacteria and the D l and D2 photosystem-I1 reaction center proteins of cyanobacteria, algae, and plants. A model of the quinone (QB) binding site of the D1 protein is presented based upon the resolved structure of the QB binding pocket of the L subunit, and introducing novel quantitative notions of complementarity and contact surface between atoms. This model, built without using traditional methods of molecular mechanics and restricted to residues in direct contact with QB, accounts for the experimentally derived functional state of mutants of the D 1 protein in the region of QB. It predicts the binding of both the classical and phenol-type PSI1 herbicides and rationalizes the relative levels of tolerance of mutant phenotypes. o 1995 Wiey-Liss, Inc.

The production of coca and poppy as well as the processing and production of cocaine and heroin involve significant environmental impacts. Both coca and poppy are grown intensively in a process that involves the clearing of land in remote areas, the planting of the crop, and protection against pests such as weeds, insects, and pathogens. The aerial spray program to control coca and poppy production in Colombia with the herbicide glyphosate is conducted with modern state-of-the-art aircraft and spray equipment. As a result of the use of best available spray and navigation technology, the likelihood of accidental off-target spraying is small and is estimated to be less than 1% of the total area sprayed. Estimated exposures in humans resulting from direct overspray, contact with treated foliage after reentry to fields, inhalation, diet, and drinking water were small and infrequent. Analyses of surface waters in five watersheds showed that, on most occasions, glyphosate was not present ...

The aim of the present study was to show the abilities of captopril as a thiol ACEi (angiotensin converting enzyme inhibitor), on mitochondria toxicity due to paraquat. Mitochondrial isolation from rat liver was divided into 4 groups. Group 1 was considered as control, group 2 received paraquat (5 mM), group 3 received captopril (0.08 mM) and group 4 received paraquat (5 mM) + captopril (0.08 mM).

The widely used herbicide paraquat was evaluated for genotoxicity in peripheral blood human lymphocyte cultures. Sister-chromatid exchanges (SCE), chromosome aberrations (CA), and micronuclei (MN) were scored as genetic endpoints. Paraquat was administered either alone or in combination with an external source of metabolic activation. Our data indicate that paraquat induced slight but significant increases in the frequency of SCE. This genotoxic effect was not modified by cotreatments with S9 fraction from rat liver. However, paraquat did not increase the frequency of CA and MN, indicating that this bipyridylium compound is not effective in these assays, which would mean a general lack of effectiveness of the herbicide to induce clastogenic damage. In addition, cotreatments with the S9 fraction, did not modify the genotoxic ability detected in the SCE assay. Teratogenesis Carcinog. Mutagen. 17:339-347, 1997/98.

Mineralisation of the phenylurea herbicide isoproturon (3-(4-isopropylphenyl)-1,1-dimethylurea) and two of its known metabolites, 3-(4-isopropylphenyl)-1-methylurea (monodesmethyl-isoproturon) and 4-isopropylaniline, was studied in Danish agricultural soils with or without previous exposure to isoproturon. A potential for rapid mineralisation of isoproturon and the two metabolites was present in soils sampled from three plots within an agricultural field previously treated regularly with the herbicide, with 34-45%, 51-58% and 33-36% of the added [phenyl-U-14 C]isoproturon, [phenyl-U-14 C]monodesmethylisoproturon and [phenyl-U-14 C]4-isopropylaniline metabolised to [ 14 C]carbon dioxide within 30 days at 20 • C. In contrast, such extensive mineralisation of these three compounds was not observed within this period in soils sampled from two other agricultural fields without previous treatment with isoproturon. The mineralisation patterns indicated growth-linked metabolism of the three compounds in the previously exposed soils, and doubling times for [ 14 C]carbon dioxide production ranged from 1.6 to 3.2, 1.0 to 2.1 and 1.3 to 1.7 days for isoproturon, monodesmethyl-isoproturon and 4-isopropylaniline, respectively. The ability to mineralise [phenyl-U-14 C]isoproturon to [ 14 C]carbon dioxide was successfully sub-cultured to a fresh mineral medium which provided isoproturon as sole source of carbon and nitrogen. One of the soils sampled from an agricultural field not previously treated with isoproturon showed accelerated mineralisation of [phenyl-U-14 C]4-isopropylaniline toward the end of the experiment, with a doubling time for [ 14 C]carbon dioxide production of 7.4 days. This study indicates that the occurrence of rapid mineralisation of the phenyl ring of isoproturon to carbon dioxide is related to previous exposure to the herbicide, which suggests that microbial adaptation upon repeated isoproturon use may occur within agricultural fields.

This paper reports the identification and functional expression of a gene that is involved in nitrate uptake in plants, a process essential for the assimilation of nitrate and the biological removal of nitrate from the soil solution. The CM1 gene of Arabidopsis, which when mutated confers resistance to the herbicide chlorate and a decrease in nitrate uptake, was isolated and found to encode a protein with 12 putative membrane-spanning segments. Injection of CM1 mRNA into Xenopus oocytes produces a nitrate-and pHdependent membrane depolarization, inward current, and nitrate uptake. These data show that the CM.7 gene encodes an electrogenic nitrate transporter. CM7 mRNA is found predominantly in roots and displays nitrate-and pH-dependent regulation.

The European Biocidal Products Directive 98/8/EC requires a risk assessment concerning possible effects of active ingredients on the environment. Biocides can be leached from treated materials exposed to outdoor use. These emissions have to be estimated and evaluated during the authorization procedure. Different immersion and irrigation tests were performed to investigate leaching of biocides from façade coatings. Several marketed formulations of textured coatings and paints spiked with a mixture of commonly used active ingredients (OIT, DCOIT, IPBC, carbendazim, isoproturon, diuron, terbutryn, and Irgarol 1051) were investigated. The emission process can be described by time-dependent functions that depend on the test conditions. The results of all test procedures confirm that leachability is related to water solubility and n-octanol-water partition coefficient of the active ingredients and that leaching of biocides from façade coatings is mainly a diffusion controlled process. Other factors like the composition of the product, availability and transport of water, concentration of active ingredients in the coatings, as well as UV-exposure of the coatings influence biocide emissions.

An optical biosensor based on vegetal cells entrapped in an inorganic translucent matrix and fluorescence detection has been developed. The biosensor uses Chlorella vulgaris immobilized in a translucent support produced from sol–gel technology. The translucence of the structure enables the algal active layer to be placed directly in contact with the optical fibers for fluorescence detection. This configuration has many advantages over the use of an opaque support because no space between the optical fibers and the active layer is required to collect fluorescence. This reagentless biosensor allows determination of diuron as an anti-PSII herbicide and its long term activity is assessed.

Arsenic (As) bioaccessibility in contaminated soils (n = 50) was assessed using the simplified bioaccessibility extraction test (SBET). Soils used in the study were collected from sites where As was used as an herbicide (railway corridor) or pesticide (cattle dip sites), from former gold mines and from highly mineralised locations containing geogenic As sources (gossans). In all but three soils, As bioaccessibility was less than 50% indicating that a significant proportion of the total As concentration may not be available for absorption in the gastrointestinal tract following incidental soil ingestion. When regression models were developed based on soil properties, the descriptive variables best able to describe As bioaccessibility in railway corridor, dip site and mine site soils were total As and total or dithionitecitrate extractable (free) iron (Fe). While As bioaccessibility could be predicted (r 2 = 0.955, n = 50) in these contaminated soils, As bioaccessibility for gossan soils was a poor fit using linear or multivariate regression analysis.

a b s t r a c t 4,5-Dichloro-1,2,3-dithiazolium chloride 2 (Appel salt) reacts with either DMSO, diphenylsulfoxide 11 or methylphenylsulfoxide 12 to give 4-chloro-5H-1,2,3-dithiazol-5-one 1 in excellent yields. The use of catalytic amounts of DMSO (1 mol %) in MeCN in the presence of water (1 equiv) transforms Appel salt 2 into dithiazolone 1 in near quantitative yields (92%). Rational mechanisms are proposed to explain the catalytic nature of these reactions.

Plants expressing Agrobacterium sp. strain CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) are known to be resistant to glyphosate, a potent herbicide that inhibits the activity of the endogenous plant EPSPS. The RR1445 transgenic cotton line (current commercial line for Roundup Ready ® Cotton) was generated using the figwort mosaic virus (FMV) 35S promoter to drive the expression of the CP4 EPSPS gene, and has excellent vegetative tolerance to glyphosate. However, with high glyphosate application rates at developmental stages later than the four-leaf stage (late-stage applications:

Anthropic changes and chemical pollution confront wild plant communities with xenobiotic combinations of bioactive molecules, degradation products, and adjuvants that constitute chemical challenges potentially affecting plant growth and fitness. Such complex challenges involving residual contamination and mixtures of pollutants are difficult to assess. The model plant Arabidopsis thaliana was confronted by combinations consisting of the herbicide glyphosate, the fungicide tebuconazole, the glyphosate degradation product aminomethylphosphonic acid (AMPA), and the atrazine degradation product hydroxyatrazine, which had been detected and quantified in soils of field margins in an agriculturally intensive region. Integrative analysis of physiological, metabolic, and gene expression responses was carried out in dose-response experiments and in comparative experiments of varying pesticide combinations. Field margin contamination levels had significant effects on plant growth and metabolism despite low levels of individual components and the presence of pesticide degradation products. Biochemical and molecular analysis demonstrated that these less toxic degradation products, AMPA and hydroxyatrazine, by themselves elicited significant plant responses, thus indicating underlying mechanisms of perception and transduction into metabolic and gene expression changes. These mechanisms may explain observed interactions, whether positive or negative, between the effects of pesticide products (AMPA and hydroxyatrazine) and the effects of bioactive xenobiotics (glyphosate and tebuconazole). Finally, the metabolic and molecular perturbations induced by low levels of xenobiotics and associated degradation products were shown to affect processes (carbon balance, hormone balance, antioxidant defence, and detoxification) that are likely to determine environmental stress sensitivity.

The joint toxicity of nonylamine and decylamine and of atrazine and decylamine was evaluated in assays with the green alga Selenastrum capricornutum based on an isobologram method. In this method, curves of constant response, isoboles, are plotted versus concentrations of two toxicants. The response parameter was growth rate based on biomass, and several response levels were used. Dose-response curves were developed for dilution series using fixed ratios between concentrations in toxic units of the compounds. Probit and Weibull dose-response curves were then determined by nonlinear regression. A model for isoboles for partially similar action was used when applicable. The no-effect concentration (NEC or EC0) was estimated based on a newly proposed model containing median effective concentration (EC50) and EC0 as explicit variables. Results show that nonylamine and decylamine are nearly concentration additive at EC50 and EC10 (similarity parameter ϭ 0.70-0.76) and to a lesser extent at EC0. By contrast, the mixtures of atrazine and decylamine show antagonism in that atrazine acts as an antidote to decylamine. The shapes of these isoboles are independent of response level. The EC50 values (mg/L) for chemicals acting singly were 0.090 (nonylamine), 0.039 to 0.044 (decylamine), and 0.225 (atrazine). In order to determine NEC effectively, the level of inhibition must be fairly low, with observed growth rates between 0.6 and 1.0 times the average growth rate of the controls.

A recent case of chronic arsenic intoxication due to prolonged accidental ingestion of a commercially available crabgrass killer illustrates one of the more common etiologies of present day arsenic poisoning. A review of 20 years of arsenic poisoning at the University of Kentucky (21 cases) revealed no cases attributable to an industrial source. A tabulation of the 21 most recent cases is included.

Few studies have been done to evaluate the transfer of catabolic plasmids from an introduced donor strain to indigenous microbial populations as a means to remediate contaminated soils. In this work we determined the effect of the conjugative transfer of two 2,4-D degradative plasmids to indigenous soil bacterial populations on the rate of 2,4-D degradation in soil. We also assessed the influence of the presence of 2,4-D on the number of transconjugants formed. The two plasmids used, pEMT1k and pEMT3k, encode 2,4-D degradative genes (tfd) that differ in DNA sequence as well as gene organisation, and confer different growth rates to Ralstonia eutropha JMP228 when grown with 2,4-D as a sole carbon source. In an agricultural soil (Ardoyen) treated with 2,4-D (100 ppm) there were ca. 10(7) CFU of transconjugants per gram bearing pEMT1k as well as a high number of pEMT3k bearing transconjugants (ca 10(6) CFU/g). In this soil the formation of a high number of 2,4-D degrading transconjugan...

Investigation on organic xenobiotics bioaccumulation/biodegradation in green algae is of great importance from environmental point of view because widespread distribution of these compounds in agricultural areas has become one of the major problems in aquatic ecosystem. Also, new technology needs to be developed for environmental detection and re-usage of the compounds as bioresources. Prometryne as a herbicide is widely used for killing annual grasses in China and other developing countries. However, overuse of the pesticide results in high risks to contamination to aquatic environments. In this study, we focused on analysis of bioaccumulation and degradation of prometryne in Chlamydomonas reinhardtii, a green alga, along with its adaptive response to prometryne toxicity. C. reinhardtii treated with prometryne at 2.5-12.5 lg L À1 for 4 d or 7.5 lg L À1 for 1-6 d accumulated a large quantity of prometryne, with more than 2 mg kg À1 fresh weight in cells exposed to 10 lg L À1 prometryne. Moreover, it showed a great ability to degrade simultaneously the cell-accumulated prometryne. Such uptake and catabolism of prometryne led to the rapid removal of prometryne from media. Physiological and molecular analysis revealed that toxicology was associated with accumulation of prometryne in the cells. The biological processes of degradation can be interpreted as an internal tolerance mechanism. These results suggest that the green alga is useful in bioremediation of prometryne-contaminated aquatic ecosystems.

The south-American cordgrass, Spartina densiflora, has become the dominant plant species on recent tidal marsh restorations in the Doñ ana National Park (SW Spain). We examined the effect of different doses of glyphosate (720-7200 g a.i. ha À1 ) and imazamox (20-68 g a.i. ha À1 ) on growth and photosynthetic apparatus of S. densiflora. Imazamox had no effect on neither on growth nor photosynthetic apparatus of S. densiflora. On the contrary, glyphosate inhibited photochemical efficiency of photosynthesis from day one. Net photosynthetic rate, stomatal conductance and photosynthetic pigments and the number of new tillers were reduced. Glyphosate at high doses (ca. 7200 g a.i. ha À1 ) could be an appropriate method of control, since it has a negative effect over the photosynthetic apparatus and growth of S. densiflora. Furthermore, glyphosate and its main metabolite, AMPA, were not extracted from the soil, since they were retained by the very high iron and aluminum oxide content of this soil.

The herbicide atrazine is one of the most commonly applied pesticides in the world. As a result, atrazine is the most commonly detected pesticide contaminant of ground, surface, and drinking water. Atrazine is also a potent endocrine disruptor that is active at low, ecologically relevant concentrations. Previous studies showed that atrazine adversely affects amphibian larval development. The present study demonstrates the reproductive consequences of atrazine exposure in adult amphibians. Atrazineexposed males were both demasculinized (chemically castrated) and completely feminized as adults. Ten percent of the exposed genetic males developed into functional females that copulated with unexposed males and produced viable eggs. Atrazineexposed males suffered from depressed testosterone, decreased breeding gland size, demasculinized/feminized laryngeal development, suppressed mating behavior, reduced spermatogenesis, and decreased fertility. These data are consistent with effects of atrazine observed in other vertebrate classes. The present findings exemplify the role that atrazine and other endocrine-disrupting pesticides likely play in global amphibian declines.

The meristematic mitotic cells of Allium cepa is an efficient cytogenetic material for chromosome aberration assay on environmental pollutants. For assessing genotoxicity of pentachlorophenol (PCP), 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-diethyl-N-(butoxymethyl) acetanilide (butachlor), 50% effective concentration (EC 50 ), c-mitosis, stickiness, chromosome breaks and mitotic index (MI) were used as endpoints of genotoxicity. EC 50 values for PCP and butachlor are 0.73 and 5.13 ppm, respectively. 2,4-D evidently induced morphological changes at higher concentrations. Some changes like crochet hooks, c-tumours and broken roots were unique to 2,4-D at 5-20 ppm. No such abnormalities were found in PCP and butachlor treated groups, however, root deteriorated and degenerated at higher concentrations (<3 ppm) in PCP. MI in 2,4-D showed a low average of 14.32% followed by PCP (19.53%), while in butachlor it was recorded 71.6%, which is near to the control value. All chemicals induced chromosome aberrations at statistically significant level. The highest chromosome aberration frequency (11.90%) was recorded in PCP at 3 ppm. Large number of c-mitotic anaphases indicated that butachlor acts as potent spindle inhibitor, whereas, breaks, bridges, stickiness and laggards were most frequently found in PCP showing that it is a potent clastogen.

Since Agent Orange with its associated TCDD contaminant was aerially disseminated on the test grids, Test Area C-52A provided a 'field laboratory' for what may have happened in Vietnam, had there been no intercepting forest cover. However, in Vietnam a 'typical' mission would have disseminated 14.8 kg of 2,4,5-T/ha, most of which was intercepted by the forest canopy, versus the 876 kg 2,4,5-T/ha on the test grid at Eglin. Moreover, each hectare on the Eglin test grid received at least 1,300 times more TCDD than a hectare sprayed with Agent Orange in Vietnam. The disappearance or persistence of TCDD is dependent upon how it enters the ecosystem. Spray equipment test and evaluations missions at Eglin were generally scheduled and conducted with environmental conditions that were optimal for spray operations. This suggests that conditions favorable for dissemination of herbicide were the same conditions favorable for photodegradation of TCDD. It was likely that 99 percent of the TCDD never persisted beyond the day of application. No long-term adverse ecological effects were documented in these studies despite the massive quantities of herbicides and TCDD that were applied to the site. Reviews by the US Environmental Protection Agency and the National Academy of Sciences' Institute of Medicine did not address the fate of Agent Orange and TCDD as described in these studies from Eglin AFB, Florida.

On/off patch spraying based on weed maps is used in site-specific weed management. Two prerequisites for realising patch spraying are accurate weed detection and targeting of the herbicides on weed patches. There is plenty in the literature about weed detection, but little attention has been paid to the spatial accuracy of herbicide application. This study was conducted in order to assess the extent to which patch-sprayed herbicides are targeted precisely according to pre-loaded prescription maps and to evaluate a new spatial assessment method. The new method consisted of spraying with red Ponceau 4R dye, unmanned aerial vehicle (UAV) imagery and spatial image analysis based on a geographic information system (GIS). The sprayed dye was clearly visible in aereal images and the locations of the sprayed areas were compared with the locations given by the prescription maps. Four different commercial sprayers with boom section width in the range of 0.5 to 3 m and driving speed in the range of 2.5 to 8 km h −1 were used in ten experiments. All the experiments were carried out in autumn in stubble fields. The results showed that the new method was fast and reliable. The incorrectly sprayed area outside targeted areas on prescription maps averaged 81% for three different sprayers with 3 m boom sections, and 5% for a sprayer with 0.5 m boom sections (individual spray nozzle control). The target areas not sprayed within the planned weed patch areas averaged 6% of the pre-defined patch area for spray-ers with 3 m boom sections, and 14% for the sprayer with 0.5 m boom sections. This study revealed that the sprayer with 0.5 m boom sections had a controller that was not quick enough at opening and closing spray nozzles at normal driving speeds. Log files from the sprayer console overestimated the sprayed area by 24% and were less accurate than the spatial analysis of the sprayed areas.

BACKGROUND: Waterhemp is a model for weed genomics research in part because it possesses many interesting biological characteristics, rapidly evolves resistance to herbicides and has a solid foundation of previous genetics work. To develop further the genomics resources for waterhemp, the transcriptome was sequenced using Roche GS-FLX 454 pyrosequencing technology.

The major herbicide used worldwide, Roundup, is a glyphosate-based pesticide with adjuvants. Glyphosate, its active ingredient in plants and its main metabolite (AMPA) are among the first contaminants of surface waters. Roundup is being used increasingly in particular on genetically modified plants grown for food and feed that contain its residues. Here we tested glyphosate and its formulation on mature rat fresh testicular cells from 1 to 10000 ppm, thus from the range in some human urine and in environment to agricultural levels. We show that from 1 to 48 h of Roundup exposure Leydig cells are damaged. Within 24-48 h this formulation is also toxic on the other cells, mainly by necrosis, by contrast to glyphosate alone which is essentially toxic on Sertoli cells. Later, it also induces apoptosis at higher doses in germ cells and in Sertoli/germ cells co-cultures. At lower non toxic concentrations of Roundup and glyphosate (1 ppm), the main endocrine disruption is a testosterone decrease by 35%. The pesticide has thus an endocrine impact at very low environmental doses, but only a high contamination appears to provoke an acute rat testicular toxicity. This does not anticipate the chronic toxicity which is insufficiently tested, and only with glyphosate in regulatory tests.

The LC/ESI/MSMS method allows the trace quantification (ng/l) of the new triketone herbicides, i.e. sulcotrione and mesotrione, and important herbicides and metabolites, in natural waters. Solid phase extraction (SPE) for sample enrichment is performed with OASIS® (recoveries 94–112% for parent herbicides). Neutral and acidic compounds were analyzed separately with ESI in positive and negative mode, respectively. Quantification limits varied between

Herbicides are still used to control weeds on hard surfaces, including municipal, private and industrial sites. Used under unfavourable conditions, especially when rain occurs shortly after application, herbicides may run off to surface waters. Such losses of herbicides from hard surfaces are estimated to be much higher than for herbicides used in arable fields. In this study, three kinds of hard surface were evaluated: asphalt, concrete surface and gravel (fine and coarse). Three herbicides were applied: glyphosate, diuron and diflufenican. Adsorption isotherms of diuron and diflufenican to the three surfaces were determined. At different times after treatment with the herbicides, rainfall was simulated by use of a rain-droplet spray nozzle, and the run-off was collected for analysis. After this run-off event, the materials were immersed in water to measure desorption which, together with the compound in the run-off, gave a measure of the dislodgable residues. The apolar herbicides diuron and especially diflufenican adsorbed strongly to asphalt. The polar herbicide glyphosate lost 75% in run-off from asphalt but was adsorbed strongly to soil and concrete pavement.

Pulse amplitude modulation (PAM) fluorometry is ideally suited to measure the sub-lethal impacts of photosystem II (PSII)-inhibiting herbicides on microalgae, but key relationships between effective quantum yield [Y(II)] and the traditional endpoints growth rate (l) and biomass increase are unknown. The effects of three PSII-inhibiting herbicides; diuron, hexazinone and atrazine, were examined on two tropical benthic microalgae; Navicula sp. (Heterokontophyta) and Nephroselmis pyriformis (Chlorophyta). The relationships between Y(II), l and biomass increase were consistent (r 2 P 0.90) and linear (1:1), validating the utility of PAM fluorometry as a rapid and reliable technique to measure sub-lethal toxicity thresholds of PSII-inhibiting herbicides in these microalgae. The order of toxicity (EC 50 range) was: diuron (16-33 nM) > hexazinone (25-110 nM) > atrazine (130-620 nm) for both algal species. Growth rate and photosynthesis were affected at diuron concentrations that have been detected in coastal areas of the Great Barrier Reef.

Plants with specific resistance to a single class of herbicides have been genetically engineered by introduction of genes encoding rationally designed mutant acetohydroxyacid synthase (AHAS) enzymes. Suitable substitution mutations were identified from a three-dimensional model of an AHAS-inhibitor complex. The structural model was generated based on homology to pyruvate oxidase and an imidazolinone inhibitor was positioned in the proposed binding site using structure-activity data for this class of herbicide. Biochemical analysis of the mutant proteins expressed in Escherichia coli enabled iterative optimization of the mutant genes. Expression of recombinant proteins in tobacco plants conferred resistance in vivo. The novel approach coupling molecular modeling and molecular biology has many advantages over traditional random mutagenesis and selection methods and will be crucial to the future development for environmentally safe and sustainable agricultural systems.

BACKGROUND: Target-site-based resistance to acetyl-CoA carboxylase (ACCase) inhibitors in Alopecurus myosuroides Huds. is essentially due to five substitutions (Isoleucine-1781-Leucine, Tryptophan-2027-Cysteine, Isoleucine-2041-Asparagine, Aspartate-2078-Glycine, Glycine-2096-Alanine). Recent studies suggested that crossresistance patterns associated with each mutation using a seed-based bioassay may not accurately reflect field resistance. The authors aimed to connect the presence of mutant ACCase isoform(s) in A. myosuroides with resistance to five ACCase inhibitors (fenoxaprop, clodinafop, haloxyfop, cycloxydim, clethodim) sprayed at the recommended field rate.

Using chlorophyll (Chl) a fluorescence many aspects of the photosynthetic apparatus can be studied, both in vitro and, noninvasively, in vivo. Complementary techniques can help to interpret changes in the Chl a fluorescence kinetics. Kalaji et al. (Photosynth Res 122:121-158, 2014a) addressed several questions about instruments, methods and applications based on Chl a fluorescence. Here, additional Chl a fluorescence-related topics are discussed again in a question and answer format. Examples are the effect of connectivity on photochemical quenching, the correction of F V /F M values for PSI fluorescence, the energy partitioning concept, the interpretation of the complementary area, probing the donor side of PSII, the assignment of bands of 77 K fluorescence emission spectra to fluorescence emitters, the relationship between prompt and delayed fluorescence, potential problems when sampling tree canopies, the use of fluorescence parameters in QTL studies, the use of Chl a fluorescence in biosensor Hazem M. Kalaji and Gert Schansker have contributed equally to this paper.

Some oenological practices, such as the massive utilisation of commercial yeast and the consequent colonisation of wineries, can contribute to reducing the native yeast biodiversity. In this context, the vineyard could be a reservoir of autochthonous yeasts of oenological interest. Thus, the evaluation of the influence of different agricultural parameters on the biodiversity of yeast population in the vineyard is necessary. This work shows the results of the influence of some floor management strategies of the vineyard in the natural yeast population associated with the grape-berries. With this objective, a three year sampling plan was designed in the Shiraz vineyards of the Madrid region using three floor management strategies: bare soil by tillage, bare soil maintained with herbicides and soil maintained with cover crop. The results of this study have shown that bare soil by tillage could be a sustainable alternative for managing the soil, due to the reduced use of agrochemicals and the resulting high yeasts biodiversity. Nevertheless, the presence of herbicides in the vineyard has a minor impact on the diversity of grape associated yeast communities, and this could have increased the yeast populations. Hence, from the fermentative yeasts' (like Saccharomyces) point of view, in hot and arid environments where soils may be affected by the tillage management, the best option could be the maintenance of the bare soil with the use of herbicides.

The triazine herbicide atrazine (2-chloro-4-ethylamino-6-isopropyl-amino-s-triazine) is one of the most used pesticides in North America. Atrazine is principally used for control of certain annual broadleaf and grass weeds, primarily in corn but also in sorghum, sugarcane, and, to a lesser extent, other crops and landscaping. Atrazine is found in many surface and ground waters in North America, and aquatic ecological effects are a possible concern for the regulatory and regulated communities. To address these concerns an expert panel (the Panel) was convened to conduct a comprehensive aquatic ecological risk assessment. This assessment was based on several newly suggested procedures and included exposure and hazard subcomponents as well as the overall risk assessment. The Panel determined that use of probabilistic risk assessment techniques was appropriate. Here, the results of this assessment are presented as a case study for these techniques. The environmental exposure assessment concentrated on monitoring data from Midwestern watersheds, the area of greatest atrazine use in North America. This analysis revealed that atrazine concentrations rarely exceed 20 g/L in rivers and streams that were the main focus of the aquatic ecological risk assessment. Following storm runoff, biota in lower-order streams may be exposed to pulses of atrazine greater than 20 g/L, but these exposures are short-lived. The assessment also considered exposures in lakes and reservoirs. The principal data set was developed by the U.S. Geological Survey, which monitored residues in 76 Midwestern reservoirs in 11 states in 1992-1993. Residue concentrations in some reservoirs were similar to those in streams but persisted longer. Atrazine residues were widespread in reservoirs (92% occurrence), and the 90th percentile of this exposure distribution for early June to July was about 5 g/L. Mathematical simulation models of chemical fate were used to generalize the exposure analysis to other sites and to assess the potential effects of reduction in the application rates. Models were evaluated, modified, and calibrated against available monitoring data to validate that these models could predict atrazine runoff. PRZM-2 overpredicted atrazine concentrations by about an order of magnitude, whereas GLEAMS underpredicted by a factor of 2 to 5. Thus, exposure models were not used to extrapolate to other regions of atrazine use in this assessment. The effects assessment considered both freshwater and saltwater toxicity test results. Phytoplankton were the most sensitive organisms, followed, in decreasing order of sensitivity, by macrophytes, benthic invertebrates, zooplankton, and fish. Atrazine inhibits photophosphorylation but typically does not result in lethality or permanent cell damage in the short term. This characteristic of atrazine required a different model than typically used for understanding the potential impact in aquatic systems, where lethality or nonreversible effects are usually assumed. In addition, recovery of phytoplankton from exposure to 5 to 20 g/L atrazine was demonstrated. In some mesocosm field experiments, phytoplankton and macrophytes were reduced after atrazine exposures greater than 20 g/L. However, populations were quickly reestablished, even while atrazine residues persisted in the water.