Growth rate effects, responses of antioxidant enzymes and metabolic fate of the herbicide Propanil in the aquatic plant Lemna minor

Propanil (3,4-dichloropropionanilide) is a selective contact pesticide, recommended for post-emergence use in rice. This herbicide may end up in surface waters and present potential risk for aquatic vascular plants. Therefore, its toxicity was evaluated on Lemna minor L., an aquatic plant regularly used for toxicological studies, during time- and concentration-dependent exposure. Toxicity assessments were based on inhibition of growth of L. minor cultures after 24 days. The obtained results showed that the growth of Lemna was affected by the herbicide. The responses of the guaiacol peroxidase (G-POD) and glutathione S-transferase (GST) involved in the xenobiotic metabolism and antioxidative system were also investigated following Propanil exposure. Our results showed that Propanil has not induced enzymatic antioxidative defenses of L. minor. Both 3,4-dichloroaniline (3,4-DCA) and 3,4-dichloroacetanilide are the major metabolites in this plant. On the contrary, only 3,4-DCA was found in culture media after 4 days. Probably, the enzymatic hydrolysis by acyl acylamidase and the acetylation by acetyl-CoA are the major pathways for these transformation products, respectively. The results of this study showed that the selected aquatic plant has the potential to accumulate and metabolize rice herbicide, like Propanil. Based on these toxicity data this herbicide should impair the establishment of non-target aquatic plants.     

Phytotoxicity and antioxidative enzymes of green microalga (Desmodesmus subspicatus) and duckweed (Lemna minor) exposed to herbicides MCPA, chloridazon and their mixtures

In this study, we evaluate the toxicity of MCPA (auxin-like growth inhibitor), chloridazon (CHD) (PSII-inhibitor) and their mixtures to floating plants and planktonic algae. Toxicity of MCPA (4-chloro-2-methylphenoxyacetic acid) and CHD (5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone) was first assessed in two growth inhibition tests with Lemna minor (ISO/DIS 20079) and Desmodesmus subspicatus (ISO 8692). Next, herbicide mixtures at concentrations corresponding to the EC values were used to assess their interactive effects, and the biomarkers were: for duckweed fresh weight, frond area, chlorophyll content and number of fronds, and for algae cell count and cell volume. The 3d EC?? and EC?? values using cell counts of D. subspicatus were 142.7 and 529.1 mg/L for MCPA and 1.7 and 5.1 mg/L for CHD. The 7d EC?? and EC?? values using frond number of L. minor amounted to 0.8 and 5.4 mg/L for MCPA and 0.7 and 10.4 mg/L for CHD. Higher sensitivity of reproductive (number of cells/fronds) than growth processes (cell volume/frond area) to herbicides applied individually and in mixtures was especially pronounced in the responses of Desmodesmus. Herbicide interactions were assessed by the two-way ANOVA and Abbott's formula. Generally, an antagonistic interaction with Lemna was revealed by MCPA and chloridazon, whereas additive effect of both herbicides was observed for Desmodesmus. A significant stimulation of SOD and APX activity by binary mixtures was noted in algal cells mainly after 24 and 48 hours of exposure. The extremely high stimulation of the activity of both enzymes was induced by the combination EC??CHD + EC??MCPA (48 h). Presumably due to oxidative stress, the treatment with CHD at concentration EC?? after 72 h was lethal for algae grown in aerated cultures, in contrast to standardized test conditions. Taking into account the consequences of risk assessment for herbicide mixtures we can state that a relatively low toxicity, as well as the lack of significant synergy between MCPA and CHD to non-target plants appears to be the most important result.     

Calcium chloride and calcium bromide aqueous solutions of technical and analytical grade in Lemna bioassay

Saturated water solutions of calcium chloride, calcium bromide and their 1:1 mixture are commonly used as "high density brines" for pressure control in oil wells. To compare the effect of these chemicals of technical grade with the effect of the chemicals of analytical grade the Lemna test was used. The multiplication rate, fresh weight, dry to fresh weight ratio, area covered by plants and chlorophyll content were measured as toxicity parameters. The concentrations of tested chemicals were 0.025, 0.05. 0.075 and 0.1 mol dm(-3). Generally, the chemicals of both technical and analytical grade in concentrations of 0.025 mol dm(-3) stimulated the Lemna minor growth, while tested chemicals in concentrations of 0.05 mol dm(-3) did not affect the growth significantly. The exceptions were results obtained by measuring fresh weight. Most of tested chemicals in concentrations of 0.075 mol dm(-3) and all chemicals in concentrations of 0.1 mol dm(-3) reduced the growth. No major differences between effects of tested chemicals of technical and analytical grade on plant growth were observed, except that tested chemicals of analytical grade in concentrations of 0.1 mol dm(-3) increased dry to fresh weight ratio much stronger than chemicals of technical grade. All tested chemicals in all concentrations increased chlorophyll content. After treatment with chemicals of analytical grade much higher increase of chlorophyll a concentration in comparison to increase of chlorophyll b was noticed, while chemicals of technical grade caused more prominent increase of chlorophyll b.     

Sorption-desorption of atrazine and diuron in soils from southern Brazil

The objective of this study was to investigate the behavior of sorption and desorption of the herbicides atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine) and diuron [3-(3,4-dichlorophenyl)-1,1-dimethyleurea] in soil samples from a typical lithosequence located in the municipality of Mamborê (PR), southern Brazil. Five concentrations of 14C-atrazine and 14C-diuron were used for both herbicides (0.48, 0.96, 1.92, 3.84, and 7.69 mg L(-1)). Sorption of both herbicides correlated positively with the organic carbon and clay content of the soil samples. Sorption isotherms were well described by the Freundlich model. The slope values of the isotherm (N) ranged from 0.84 to 0.90 (atrazine) and from 0.75 to 0.79 (diuron) for the lithosequence samples. Sorption of diuron was high regardless of the soil texture or the concentration added. The desorption isotherms for atrazine and diuron showed good fit to the Freundlich equation (R2 >or= 0,87). Atrazine slope values for the desorption isotherms were similar for the different concentrations and were much lower than those observed for the sorption isotherms. Significant hysteresis was observed in the herbicide desorption. When the two herbicides were compared, it was found that diuron (N = 0.06-0.22) presented more pronounced hysteresis than atrazine. The results showed that, quantitatively, a greater atrazine fraction applied to these soils remains available to be leached in the soil profile, as compared to diuron.     

Comparative phytotoxicity of Rac-metolachlor and S-metolachlor on rice seedlings

The toxicity of Rac-metolachlor and S-metolachlor on rice (IIYou92, Oryza sativa L.) seedlings was determined and compared in a hydroponics experiment. The elongation of shoot, main root, and the number of lateral roots were inhibited in both Rac- and S-metolachlor treatments. The 96 h- IC50 of Rac- and S-metolachlor were 12.32 and 9.44 μM for shoot, and 4.69 and 1.75 μM for root, respectively. The content of chlorophyll a, chlorophyll b and total chlorophyll of leaf treated by Rac-metolachlor was higher than that treated by S-metolachlor. The activities of superoxide dismutase, peroxidase, cytochrome P450 2E1, glutathione S-transferase, and the content of glutathione increased after herbicide exposure, and the activities or the content was higher in the S-metolachlor treatment than in the exposure to Rac-metolachlor. Ultrastructural studies revealed that Rac- and S-metolachlor had adverse effects on leaf cells, and S-metolachlor treatment caused higher damage.     

Temporal changes in the toxicity of pentachlorophenol to Chlorella pyrenidosa algae

The toxicity of pentachlorophenol (PCP) on Chlorella pyrenidosa algae was investigated with specific attention given to possible variation of toxic effects with time. A concentration-effect relationship was observed in which there was significant inhibition of PCP on cell density and chlorophyll A content. The inhibition rate of PCP on cell density was dependent on exposure time. The IC50 values after exposure times of 2, 4 and 6 days for cell growth were 4.18 +/- 0.49, 3.49 +/- 0.40 and 3.30 +/- 0.26 mg/L, respectively. There was also inhibition of chlorophyll A production, which appeared to increase marginally with exposure time for a given concentration of PCP. The corresponding IC50 values on day 2, 4 and 6 were 2.30 +/- 0.12, 2.63 +/- 0.38 and 3.30 +/- 0.34 mg/L, respectively. The effect of PCP on nitrate reductase (NR), was first stimulation followed by an inhibition phase. It is postulated that the observed temporal changes in the activity of nitrate reductase (NR) may occur through the addition or loss of phosphorus in the NR protein.     

Effects of procymidone, fludioxonil and pyrimethanil on two non-target aquatic plants.

Procymidone, fludioxonil, and pyrimethanil are widely used to control the pathogenic fungus Botrytis cinerea in Champagne's vineyards. These fungicides may end up in surface waters and present potential risks for aquatic vascular plants and algae. Therefore, their toxicity was evaluated on Lemna minor and Scenedesmus acutus in six-day or 48-h tests, respectively. Based on growth and chlorophyll (Chl) content of L. minor and S. acutus cultures, the results showed that the alga was the most sensitive to the fungicides. Among the fungicides, pyrimethanil was the most toxic for L. minor, its nominal IC50 was 46.16 mg l(-1) and that of the other two was >100 mg l(-1). In contrast, pyrimethanil appeared the least toxic for S. acutus at low concentration, nominal IC50 were 22.81, 4.85, and 4.55 mg l(-1) for pyrimethanil, fludioxonil, and procymidone, respectively. Fate of the fungicides in the media was also investigated and acute toxicity of the agrochemicals is discussed in regard to concentration in the culture media. Poor solubility of procymidone and fludioxonil appeared to be partly responsible for the low toxicity of these fungicides. Based on these toxicity data and the concentrations found in ponds collecting vineyard runoff water, these pesticides should not impair the establishment of pioneer plants.     

Impact of four pesticides on the growth and metabolic activities of two photosynthetic algae

The acute toxicity was determined for soil algae Chlorella kesslerei and Anabaena inaequalis, exposed to pesticides lindane, pentachlorophenol (PCP), isoproturon (IPU), and methyl parathion (MP). Toxicity markers included growth inhibition, chlorophyll biosynthesis, and total carbohydrate content, as a function of dose and time. Concentration response functions (EC50) were estimated by probit data transformation and weighted linear regression analyses. Lindane's toxicity to Chlorella increased sharply with time (EC50 = 7490, 10.3, 0.09 mg L(-1); 24, 48, 72 h), but remained nearly constant through 72 h with Anabaena (8.7-6.7 mg L(-1); 24-72 h). PCP at low concentrations stimulated algal growth and chlorophyll a production, an effect reversed at higher doses. Anabaena was less tolerant of PCP and MP than was Chlorella. The 96-h static EC50 values for Chlorella were: 0.003, 34, 0.05, and 291 mg L(-1) for lindane, PCP, isoproturon, and MP, respectively; for Anabaena, these were 4.2, 0.13, 0.21, and 19 mg L(-1). Carbohydrate production responses were similar to those of cell density (growth) and chlorophyll biosynthesis, with MP having the lowest adverse impact. The overall relative toxicity among the four tested pesticides was: for Chlorella, lindane > IPU > PCP > MP; and for Anabaena, PCP > IPU > lindane > MP. The results confirm that toxicants such as these pesticides may affect individual (though related) species to significantly different degrees.     

Study of the toxicity of diuron and its metabolites formed in aqueous medium during application of the electrochemical advanced oxidation process "electro-Fenton"

Diuron (N'-[3,4-dichlorophenyl]-N,N-dimethylurea) is a herbicide belonging to the phenylurea family, widely used to destroy weeds on uncultivated surfaces. Because of its toxicity for aquatic organisms and suspicion of being carcinogenic for humans, diuron is the object of growing environmental concern. Therefore, we have developed the electro-Fenton method, an electrochemical advanced oxidation process (EAOP), to degrade diuron in aqueous medium, and we have studied the evolution of the toxicity of treated solution during the process. Indeed, the EAOPs catalytically generate hydroxyl radicals that oxidize the persistent organic pollutants, and can ultimately destroy and mineralize them. But, sometimes, relatively toxic organic metabolites are formed during the oxidation reaction. In this work, the evolution of toxicity of diuron aqueous solutions was studied at different initial concentrations, during treatment by the electro-Fenton method. Samples were collected at various electrolysis times and mineralization degrees during the treatment. The toxicity of the samples was measured using the bacteria Vibrio fischeri (Microtox) and the green alga Scenedesmus obliquus. Our results demonstrated that the toxicity of diuron aqueous solutions (concentrations=3.0-27.6 mg L(-1)) varied considerably with time. The formation and disappearance of several metabolites, having toxicity often stronger than that of the initial herbicide, were observed. To improve the efficiency of water decontamination, the electro-Fenton method should be applied during a time long enough (several hours) and at relatively high electrolysis current (I=250 mA) to reach a nearly complete mineralization of the herbicide in the aqueous medium.     

Comparison of different physiological parameter responses in Lemna minor and Scenedesmus obliquus exposed to herbicide flumioxazin

The sensitivity of different physiological parameters in Scenedesmus obliquus and Lemna minor exposed to herbicide (flumioxazin) was investigated to indicate the most convenient and sensitive parameter. To assess toxicity of flumioxazin, we used a panel of biomarkers: pigment contents, chlorophyll fluorescence parameters and antioxidative enzyme activities. Algae and duckweed were exposed to 48-h IC50 for growth rate. In L. minor, the sensitivity of the parameters was as follows: QN > Oxygen emmision > phiS(PSII) > QP > phi(PSII) > CAT, GR > Pigment> APO > Growth. For S. obliquus, this ranking was as follows: CAT > Oxygen emission > QP > APO > GR > Pigment > phiS(PSII) > Growth > phi(PSII) > QN (from the greatest to the least sensitive). The results demonstrated that the observed toxicity is related not only to interspecific variations but also to the selected parameter.     

Leaching of oryzalin and diuron through undisturbed vineyard soil columns under outdoor conditions

Field studies monitoring herbicide pollution in the vineyards of Burgundy (France) have revealed that drinking water reservoirs are contaminated with several pre-emergence herbicides. An assessment of the leaching of two such herbicides, diuron and oryzalin, was therefore performed using lysimeters, under outdoor conditions, from May 2001 to May 2002. Four vineyard soils from Vosne-Romanée (Burgundy) were chosen along a topolithosequence: a rendosol and three calcosols. After 673 mm of rainfall, greater amounts of diuron than oryzalin were measured in percolates: respectively 0.10-0.84% and 0.02-0.43% of applied herbicide, depending on soils. Measurements for diuron metabolites detected greater amounts of DCPMU than DCPU in the percolates: respectively 0.05-0.13% and 0-0.04% of the applied diuron. At the end of the monitoring period, more residues of diuron than oryzalin were recovered in the soil profiles: respectively 4.6-9% and 1.4-4.4%. The oryzalin residues were found mainly in the upper 10 cm of soil columns, whereas diuron residues were present in the whole core. The mobility of both oryzalin and diuron seems fairly well-related to soil organic carbon content; the mobility of diuron is also related to soil texture (sand and coarse material contents). Under such experimental conditions, this study confirms that diuron leaching, and therefore potential groundwater contamination, is greater than that of oryzalin.     

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.     

Absorption and translocation of sulfometuron-methyl in sugarcane (Saccharum officinarum L.) at different growth stages

ABSTRACT

In Brazil, weed management in sugarcane fields is mainly done with the use of selective herbicide formulations. For many years, diuron+hexazinone was one of the main herbicide mixture formulations used in sugarcane. Later, sulfometuron-methyl was included in the same mixture, which was marketed as a new herbicide formulation for residual in-season weed control in sugarcane. The mixture diuron+hexazinone+sulfometuron-methyl has been widely used in commercial sugarcane fields in Brazil. However, recent field observations have shown that sugarcane plants at different growth stages varied in their phytotoxicity levels after treatment with diuron+hexazinone+sulfometuron-methyl. Greenhouse and laboratory studies were conducted to determine ¹⁴Csulfometuron-methyl absorption and translocation, as well as ¹⁴C distribution in sugarcane at two growth stages, 2 to 3 leaves and 5 to 6 leaves. ¹⁴Csulfometuron-methyl absorption by sugarcane did not differ between the two growth stages. Different patterns of ¹⁴C accumulation were observed, which may explain variations in sulfometuron-methyl phytotoxic responses observed in the field.     

Toxicity and removal of pesticides by selected aquatic plants

Pesticides are being detected in water bodies on an increasingly frequent basis. The present study focused on the phytoremediation potential of selected aquatic plants to remove phytosanitary products from contaminated water. We investigated the uptake capacity of Lemna minor (L. minor), Elodea canadensis (E. canadensis) and Cabomba aquatica (C. aquatica) on three pesticides: copper sulphate (fungicide), flazasulfuron (herbicide) and dimethomorph (fungicide). Pesticide toxicity was evaluated by exposing plants to five concentrations (0-1 mg L(-1)) in culture media for 7d using chlorophyll fluorescence as a biomarker. The toxicity of the contaminants was the same for all the aquatic plants studied and occurred in this descending order of toxicity: flazasulfuron>copper>dimethomorph. We found that L. minor had the most efficient uptake capacity, followed by E. canadensis and then C. aquatica. The maximum removal rate (microg g(-1)fresh weight d(-1)) of copper, flazasulfuron and dimethomorph was 30, 27 and 11, respectively.     

Detection of early effects of a single herbicide (diuron) and a mix of herbicides and pharmaceuticals (diuron, isoproturon, ibuprofen) on immunological parameters of Pacific oyster (Crassostrea gigas) spat

In the context of massive summer mortality events of the Pacific oyster Crassostrea gigas, the aim of this study was to investigate the early effects on genes, enzymes and haemocyte parameters implicated in immune defence mechanisms in C. gigas oysters exposed to a potentially hostile environment, i.e. to an herbicide alone or within a mixture. Following 2 h of exposure to the herbicide diuron at 1 μg L⁻¹, the repression of different genes implicated in immune defence mechanisms in the haemocytes and the inhibition of enzyme activities, such as laccase-type phenoloxidase (PO) in the plasma, were observed. The inhibition of superoxide dismutase (SOD) activity in the plasma was also observed after 6 and 24 h of exposure. In the mixture with the herbicides diuron and isoproturon, and the pharmaceutical ibuprofen, catecholase-type PO activity in the plasma and the percentage of phagocytosis in the haemocytes were reduced after 6 h of exposure. Our results showed that early effects on molecular, biochemical and cellular parameters can be detected in the presence of diuron alone or within a mixture, giving an insight of its potential effect in situations that can be found in natural environments, i.e. relatively high concentrations for short periods of time.     

Diuron mobility through vineyard soils contaminated with copper

The herbicide diuron is frequently applied to vineyard soils in Burgundy, along with repeated treatments with Bordeaux mixture (a blend of copper sulfate and calcium hydroxide) that result in elevated copper concentrations. Cu could in principle affect the fate and transport of diuron or its metabolites in the soil either directly by complexation or indirectly by altering the populations or activity of microbes involved in their degradation. To assess the effect of high Cu concentrations on diuron transport, an experiment was designed with ten undisturbed columns of calcareous and acidic soils contaminated with 17--509 mg kg(-1) total Cu (field-applied). Grass was planted on three columns. Diuron was applied to the soils in early May and in-ground lysimeters were exposed to outdoor conditions until November. Less than 1.2% of the diuron applied was found in the leachates as diuron or its metabolites. Higher concentrations were found in the effluents from the grass-covered columns (0.1--0.45%) than from the bare-soil columns (0.02--0.14%), and they were correlated with increases in dissolved organic carbon. The highest amounts of herbicide were measured in acidic-soil column leachates (0.98--1.14%) due to the low clay and organic matter contents of these soils. Cu also leached more readily through the acidic soils (32.8--1042 microg) than in the calcareous soils (9.5--63.4 microg). Unlike in the leachates, the amount of diuron remaining in the soils at the end of the experiment was weakly related to the Cu concentrations in the soils.     

Individual and mixture toxicity of three pesticides; atrazine,chlorpyrifos, and chlorothalonil to the marine phytoplankton species Dunaliella tertiolecta

This study analyzed the toxicity of three pesticides (the herbicide atrazine, the insecticide chlorpyrifos and the fungicide chlorothalonil) individually, and in two mixtures (atrazine and chlorpyrifos; atrazine and chlorothalonil) to the marine phytoplankton species Dunaliella tertiolecta (Chlorophyta). A standard 96 h static algal bioassay was used to determine pesticide effects on the population growth rate of D. tertiolecta. Mixture toxicity was assessed using the additive index approach. Atrazine and chlorothalonil concentrations > or = 25 microg/L and 33.3 microg/L, respectively, caused significant decreases in D. tertiolecta population growth rate. At much higher concentrations (> or = 400 microg/L) chlorpyrifos also elicited a significant effect on D. tertiolecta population growth rate, but toxicity would not be expected at typical environmental concentrations. The population growth rate EC50 values determined for D. tertiolecta were 64 microg/L for chlorothalonil, 69 microg/L for atrazine, and 769 microg/L for chlorpyrifos. Atrazine and chlorpyrifos in mixture displayed additive toxicity, whereas atrazine and chlorothalonil in mixture had a synergistic effect. The toxicity of atrazine and chlorothalonil combined was approximately 2 times greater than that of the individual chemicals. Therefore, decreases in phytoplankton populations resulting from pesticide exposure could occur at lower than expected concentrations in aquatic systems where atrazine and chlorothalonil are present in mixture. Detrimental effects on phytoplankton population growth rate could impact nutrient cycling rates and food availability to higher trophic levels. Characterizing the toxicity of chemical mixtures likely to be encountered in the environment may benefit the pesticide registration and regulation process.     

Phytotoxicity and antioxidative enzymes of green microalga (Desmodesmus subspicatus) and duckweed (Lemna minor) exposed to herbicides MCPA,chloridazon and their mixtures

In this study, we evaluate the toxicity of MCPA (auxin-like growth inhibitor), chloridazon (CHD) (PSII-inhibitor) and their mixtures to floating plants and planktonic algae. Toxicity of MCPA (4-chloro-2-methylphenoxyacetic acid) and CHD (5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone) was first assessed in two growth inhibition tests with Lemna minor (ISO/DIS 20079) and Desmodesmus subspicatus (ISO 8692). Next, herbicide mixtures at concentrations corresponding to the EC values were used to assess their interactive effects, and the biomarkers were: for duckweed fresh weight, frond area, chlorophyll content and number of fronds, and for algae cell count and cell volume. The 3d EC₁₀ and EC₅₀ values using cell counts of D. subspicatus were 142.7 and 529.1 mg/L for MCPA and 1.7 and 5.1 mg/L for CHD. The 7d EC₁₀ and EC₅₀ values using frond number of L. minor amounted to 0.8 and 5.4 mg/L for MCPA and 0.7 and 10.4 mg/L for CHD. Higher sensitivity of reproductive (number of cells/fronds) than growth processes (cell volume/frond area) to herbicides applied individually and in mixtures was especially pronounced in the responses of Desmodesmus. Herbicide interactions were assessed by the two-way ANOVA and Abbott's formula. Generally, an antagonistic interaction with Lemna was revealed by MCPA and chloridazon, whereas additive effect of both herbicides was observed for Desmodesmus. A significant stimulation of SOD and APX activity by binary mixtures was noted in algal cells mainly after 24 and 48 hours of exposure. The extremely high stimulation of the activity of both enzymes was induced by the combination EC₁₀CHD + EC₅₀MCPA (48 h). Presumably due to oxidative stress, the treatment with CHD at concentration EC₅₀ after 72h was lethal for algae grown in aerated cultures, in contrast to standardized test conditions. Taking into account the consequences of risk assessment for herbicide mixtures we can state that a relatively low toxicity, as well as the lack of significant synergy between MCPA and CHD to non-target plants appears to be the most important result.     

Effects of thiobencarb herbicide to an alga (Nannochloris oculata) and the cladoceran (Daphnia magna)

Chronic toxicity studies were conducted with an algae (Nannochloris oculata) and the cladoceran (Daphnia magna) to determine their relative sensitivities to the thiocarbamate herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate). Most of the algal populations were initially affected by exposure to the herbicide. Thiobencarb concentrations higher than 0.5 mg/L significantly reduced algal densities after 24-h exposure. The 24-h static EC50 in D. magna was 3.01 mg/L. The sublethal effects of 0.3, 0.37, 0.5, 0.75, and 1.5 mg/L of thiobencarb concentrations on the survival, reproduction, and growth of D. magna were monitored for 21 days. The parameters used to determined the effect of the herbicide on D. magna were mean total young per female; mean brood size; days to first brood; intrinsic rate of natural increase (r); growth; and survival. Reproduction was significantly reduced at thiobencarb concentrations of 0.30 mg/L and higher while survival was affected after exposure to 0.75 and 1.5 mg/L of the pesticide. The r value decreased with increasing concentrations of thiobencarb. Growth, as measured by body length, was depressed significantly after exposure to all herbicide concentrations tested.     

Sorption-Desorption of Atrazine and Diuron in Soils from Southern Brazil

The objective of this study was to investigate the behavior of sorption and desorption of the herbicides atrazine (6-chloro-N ²-ethyl-N ⁴-isopropyl-1,3,5-triazine-2,4-diamine) and diuron [3-(3,4-dichlorophenyl)-1,1-dimethyleurea] in soil samples from a typical lithosequence located in the municipality of Mamborê (PR), southern Brazil. Five concentrations of ¹⁴C-atrazine and ¹⁴C-diuron were used for both herbicides (0.48, 0.96, 1.92, 3.84, and 7.69 mg L^?1). Sorption of both herbicides correlated positively with the organic carbon and clay content of the soil samples. Sorption isotherms were well described by the Freundlich model. The slope values of the isotherm (N) ranged from 0.84 to 0.90 (atrazine) and from 0.75 to 0.79 (diuron) for the lithosequence samples. Sorption of diuron was high regardless of the soil texture or the concentration added. The desorption isotherms for atrazine and diuron showed good fit to the Freundlich equation (R ² ≥ 0,87). Atrazine slope values for the desorption isotherms were similar for the different concentrations and were much lower than those observed for the sorption isotherms. Significant hysteresis was observed in the herbicide desorption. When the two herbicides were compared, it was found that diuron (N = 0.06–0.22) presented more pronounced hysteresis than atrazine. The results showed that, quantitatively, a greater atrazine fraction applied to these soils remains available to be leached in the soil profile, as compared to diuron.