Enantioselective metabolism and cytotoxicity of the chiral herbicide ethofumesate in rat and chicken hepatocytes

We investigated the metabolic kinetics and toxicity of ethofumesate (ETO) in rat and chicken hepatocytes using a chiral high-performance liquid chromatographic (HPLC) method. The metabolic of ETO in rat hepatocytes was enantioselective, whereas it was not in chicken hepatocytes. The T_1/2 of (−)-ETO was about two times longer than that of (+)-ETO after the rat hepatocytes had been incubated with 20 μM rac-ETO. There was no chiral conversion or transformation during their incubation with the hepatocytes. Toxicity differences were observed between the two enantiomers of ETO, reflected in their EC₅₀ values in rat and chicken hepatocytes. The stereoselective cytotoxicity of the two enantiomers was opposite in rat and chicken hepatocytes. We have developed a method of studying the toxicokinetics and cytotoxicity of chiral agrochemicals in hepatocytes isolated from mammals (rats) and chicken. The data presented here allow a more thorough understanding of this pesticide and should be useful in its full environmental assessment.     

Enantioselective degradation kinetics of metalaxyl in rabbits

Metalaxyl [methyl-N-(2′-methoxyacetyl)-N-(2,6-dimethylphenyl)-d,l- alaninate] is a potent phenylamide fungicide. The (−)-(R)-isomer accounts for most of the fungicidal activity. A possible stereo and/or enantioselective kinetics of metalaxyl in rabbits was investigated by intravenous injection. The concentrations of (−)-(R)- and (+)-(S)-metalaxyl in plasma, liver, and kidney tissue were determined by HPLC with a cellulose-Tris-(3,5-dimethylphenylcarbamate)-based chiral stationary phase and gas chromatography-mass spectroscopy. After intravenous administration of racemic metalaxyl (40 mg/kg), the (+)-(S)-enantiomer levels in plasma, liver, and kidney decreased more rapidly than the (−)-(R)-isomer. The area ratio of the (−)-(R)-/(+)-(S)-enantiomer under the concentration-time curve (AUC_0 → ∞) in plasma after drug application was 1.62. The total plasma clearance value of the (+)-(S)-enantiomer was 1.53 and higher than that of the (−)-(R)-enantiomer. The [R]/[S] ratio in plasma was >1 for standard rac-metalaxyl at each time point. The other pharmacokinetic parameters of the enantiomers were also different. The results indicate substantial stereoselectivity in the degradation of metalaxyl enantiomers in rabbits.     

Development of an enzyme-linked immunosorbent assay for quantitative determination of cyhalofop-butyl

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for cyhalofop-butyl was developed with a polyclonal antibody produced against a hapten (cyhalofop acid) conjugated with bovine serum albumin (BSA). The ELISA of cyhalofop-butyl showed an IC₅₀ value of 0.067 ± 0.004 mg/l and the limit of detection (LOD, IC₁₀) of 0.0029 ± 0.0001 mg/l at the optimal conditions. No significant cross-reaction to other structure-related compounds suggested high specificity for cyhalofop-butyl of the method. The average recoveries of cyhalofop-butyl from fortified water and soil were in the range of 83.2-119.7% and 80.1-104.0%, respectively. These data indicate that this method is a convenient analytical technique for monitoring cyhalofop-butyl in water and soil without purification steps.     

Persistence and movement of ethofumesate in soil*

Persistence of ethofumesate [(±)2-ethoxy-2.3-dihydro-3,3-dimethylbenzofuran-5-yl-methansulphonate] in soil was associated with soil temperature. Ethofumesate applied at 4.5 kg/ha in November persisted about twice as long in soil as that applied the following March. In another field study, 88–91% of the herbicide had dissipated after 24 weeks in sandy loam soil, compared to 72–77% in loam soil when it was applied at rates of 2.2, 3.4, 4.5, and 9.0 kg/ha. The rate of degradation was independent of the initial rate of chemical applied. The time required for 50% of the herbicide to dissipate in sandy loam and loam soils was 7.7 and 12.6 weeks, respectively. The movement of ethofumesate in these two soils over a 24-weeks sampling period was confined mainly to the upper 7.5 cm of the soil profile.     

Single and simultaneous exposure of acetylcholinesterase to diazinon, chlorpyrifos and their photodegradation products

In vitro inhibition of electric eel acetylcholinesterase (AChE) by single and simultaneous exposure to organophosphorus insecticides diazinon and chlorpyrifos, and their transformation products, formed due to photoinduced degradation, was investigated. Increasing concentrations of diazinon, chlorpyrifos and their oxidation products, diazoxon and chlorpyrifos-oxon, inhibited AChE in a concentration-dependent manner. IC₅₀ (20 min) values, obtained from the inhibition curves, were (in mol/l): (5.1 ± 0.3) × 10⁻⁸, (4.3 ± 0.2) × 10⁻⁶ and (3.0 ± 0.1) × 10⁻⁸ for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively, while maximal diazinon concentration was lower than its IC₅₀ (20 min). Calculated K_I values, in mol/l, of 7.9 × 10⁻⁷, 9.6 × 10⁻⁶ and 4.3 × 10⁻⁷ were obtained for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively. However, 2-isopropyl-4-methyl-6-pyrimidinol (IMP) and 3,5,6-trichloro-2-pyridinol, diazinon and chlorpyrifos hydrolysis products, did not noticeably affect the enzyme activity at all investigated concentrations. Additive inhibition effect was achieved for lower concentrations of the inhibitors (diazinon/diazoxon ?1 × 10⁻⁴/1 × 10⁻⁸ mol/l i.e., chlorpyrifos/chlorpyrifos-oxon ?2 × 10⁻⁶/3 × 10⁻⁸ mol/l), while an antagonistic effect was obtained for all higher concentrations of the organophosphates. Inhibitory power of 1 × 10⁻⁴ mol/l diazinon irradiated samples can be attributed mostly to the formation of diazoxon, while the presence of non-inhibiting photodegradation product IMP did not affect diazinon and diazoxon inhibitory efficiencies.     

Studies on glutathione transferase from grasshopper (Zonocerus variegatus)

Glutathione transferase (GST) was purified from the hindgut of grasshopper (Zonocerus variegatus) a polyphagous insect. The purified enzyme had a native molecular weight of 40 kDa and a subunit molecular weight of 19 kDa. The purified enzyme could conjugate glutathione (GSH) with 1-chloro-2,4-dinitrobenzene (CDNB), paranitrobenzylchloride, paranitrophenylacetate, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBDCl), and 1,2-dichloro-4-nitrobenzene (DCNB) with specific activities of 3.3 ± 0.3, 0.49 ± 0.10, 0.10 ± 0.002, 1.2 ± 0.2, and 1.7 ± 0.4 μmol/min/mg protein, respectively. CDNB appears to be the best substrate with a specificity constant, k_cat/K_m, of 1.8 ± 0.1 × 10⁻⁴ M⁻¹ S⁻¹. The kinetic mechanism of Z. variegatus GST (zvGST) in the conjugation of GSH with some electrophilic substrates appears complex. Conjugation of GSH with DCNB was inhibited by high DCNB concentration, while with NBDCl, as the electrophilic substrates, different values of K_m were obtained at high and low concentrations of the substrates. Cibacron blue, hematin, S-hexylglutathione, and oxidized glutathione inhibited the enzyme with I₅₀ values of 0.057 ± 0.004, 0.80 ± 0.2, 33 ± 2 μM, and 5.2 ± 0.3 mM, respectively. The nature of inhibition by each of these inhibitors is either competitive or non-competitive at varying GSH or CDNB as substrates. NADH and NAD⁺ inhibited the enzyme with an I₅₀ value of 0.4 ± 0.01 and 11 ± 1 mM, respectively. NADH at a concentration of 0.54 mM completely abolished the activity. As part of its adaptation, the flexible kinetic pathway of detoxication by zvGST may assist the organism in coping with various xenobiotics encountered in its preferred food plants.     

Binding of phenthoate to bovine serum albumin and reduced inhibition on acetylcholinesterase

Organophosphorus pesticides (OPs) are of environmental significance due to their high toxicity to animals. Binding to plasma proteins may effective influence the toxicological properties of xenobiotics. In an attempt to evaluate the affinity of phenthoate (PTA) to bovine serum albumin (BSA) and inhibitory ability of bound PTA to acetylcholinesterase (AChE), we investigated the interactions between phenthoate (PTA) and bovine serum albumin (BSA) using tryptophan fluorescence quenching and subsequent inhibition on AChE activity by PTA. The results showed that PTA caused the fluorescence quenching of BSA because of the formation of a PTA-BSA complex. Quenching constants (K_sv), determined using the Sterns-Volmer equation to provide a measure of the binding affinity between PTA and BSA at 303, 306, 310 and 313 K were (3.4295 ± 0.0763) × 10⁻⁴, (3.2446 ± 0.0635) × 10⁻⁴, (3.0434 ± 0.0856) × 10⁻⁴ and (2.8262 ± 0.0569) × 10⁻⁴ M⁻¹, respectively. The thermodynamic parameters, ΔH and ΔS were −25.04 kJ mol⁻¹ and 168.94 J mol⁻¹ K⁻¹, respectively, which indicated that the electrostatic interactions played a major role in PTA-BSA association. The presence of BSA consistently reduced the inhibitory ability of PTA on AChE, with the relative activity being increased from 46.98 to 61.71% for the concentration range of BSA between 0 and 4.0 g L⁻¹.     

Avermectin exposure induces apoptosis in King pigeon brain neurons

The effect of avermectin was studied on King pigeon brain nerve cells by cytotoxicity [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide, MTT] and apoptosis [acridine orange/ethidium bromide (AO/EB) assay, transmission electron microscope (TEM) evaluation, measurement of mitochondrial membrane potential (Δψm), phosphatidylserine (PS) exposure, caspases activities, DNA fragmentation, reactive oxygen species (ROS) and caspase-3 mRNA expression] within the 2.5–10 μg L⁻¹ concentration-range. The results revealed that within the concentrations of 2.5–10 μg L⁻¹, avermectin showed obvious cytotoxicity and induced apoptosis in a dose-dependent manner to neurons of King pigeon in vitro. Cell viability were 99.93 ± 8.52%, 82.02 ± 4.99% and 78.23 ± 5.67% after 24 h of treatment with avermectin at the concentrations of 0, 2.5 and 5 μg L⁻¹, which decreased to 56.36 ± 2.17% of 10 μg L⁻¹. Treated cells showed typical apoptosis morphological changes including cytoplasmic vacuolation, chromatin condensation, unclear nuclear membrane and decreased/swollen mitochondria. Typical biochemical hallmarks of apoptosis including Δψm loss, PS exposure, activations of caspase-3, caspase-8 and caspase-9, DNA fragmentation were observed too. Moreover, the levels of ROS in the avermectin treatment groups increased significantly compared to control group. Furthermore, the caspase-3 mRNA levels increased significantly following AVM treatment. In conclusion, our experimental results show that avermectin has cytotoxicity to brain neurons of King pigeon in vitro and the mechanism of neurotoxicity induced by avermectin is closely related to apoptosis.     

The toxic effects of pyrethroid deltamethrin on the common carp (Cyprinus carpio L.) embryos and larvae

Deltamethrin, a synthetic pyrethroid pesticide contaminating aquatic ecosystems as a pollutant, was investigated in the present study for toxic effects on embryos and larvae of common carp, Cyprinus carpio as a model. The control and five test experiments were repeated five times. The water temperature in the experimental units was kept at 24 ± 1 °C. The number of dead embryos significantly increased in response to deltamethrin concentrations 0.005, 0.05, 0.5, 5, 25, and 50 μg L⁻¹ (p<0.05 for each cases). Dose-response decreases in hatching success were recorded as 75.2, 64.6, 47.4, 26.0, 14.4, and 9.0%, respectively. The lowest concentration of deltamethrin (0.005 μg L⁻¹) produced a significantly decrease in number of dead larvae compared to control group (p<0.05). With increasing deltamethrin concentrations, the larvae exposed duration 1-48 h significantly increased the number of dead larvae (p<0.05 for each cases). The 48 h LC₅₀ values (with 95% confidence limits) of deltamethrin for common carp embryos and larvae were estimated as 0.213 (0.103-0.404) and 0.074 (0.011-0.260) μg L⁻¹, respectively. The results provide evidence that deltamethrin pollution may have an adverse effect on the reproduction and development of carp, which should be considered when this chemical is used in agricultural areas near aquatic ecosystems.     

Ameliorative effects of Mesorhizobium sp. MRC4 on chickpea yield and yield components under different doses of herbicide stress

The present study was conducted to assess the plant growth promoting activities of Mesorhizobium sp. in the presence of technical grade herbicides and its ameliorating effects on herbicide toxicity to chickpea grown in herbicide treated soils. The quizalafop-p-ethyl and clodinafop-tolerant Mesorhizobium isolate MRC4 recovered from the nodules of chickpea plants significantly produced IAA, siderophores, hydrogen cyanide and ammonia in medium amended with or without technical grade quizalafop-p-ethyl and clodinafop. Quizalafop-p-ethyl at 40, 80 and 120 μg kg⁻¹ soil and clodinafop at 400, 800 and 1200 μg kg⁻¹ soil in general, decreased the growth attributes of chickpea plants inoculated with Mesorhizobium MRC4 and un-inoculated chickpeas. The three concentrations of quizalafop-p-ethyl were comparatively more toxic and substantially decreased biomass, nodulation and leghaemoglobin content, nutrient uptake, seed yield and grain protein over the un-inoculated chickpea. Interestingly, Mesorhizobium isolate MRC4 with any concentration of the two herbicides significantly increased the measured parameters when compared to the plants grown in soils treated solely (without inoculant) with similar concentration of each herbicide. Conclusively, Mesorhizobium isolate MRC4 could be exploited as bio-inoculant for facilitating chickpea growth under herbicide stress.     

Acute toxicity of organophosphorous pesticide diazinon and its effects on behavior and some hematological parameters of fingerling European catfish (Silurus glanis L.)

Diazinon is commonly used for pest control in the agricultural fields surrounding freshwater reservoirs. So this study was conducted to determine the acute toxicity of this organophosphorous pesticide, contaminating aquatic ecosystems as a pollutant, and its effects on behavior, and some hematological parameters of fingerling European catfish, Silurus glanis. Diazinon was applied at concentrations of 1, 2, 4, 8, 16, 32, and 64 mg L⁻¹. The water temperature in the experimental units was kept at 16 ± 1 °C. The number of dead fishes significantly increased in response to diazinon concentrations 2-64 mg L⁻¹ (p < 0.05). With increasing diazinon concentrations, the fishes exposed duration 1 to 96 h significantly increased the number of dead fishes (p < 0.05 for each cases). The 1, 24, 48, 72, and 96 h LC₅₀ values (with 95% confidence limits) of diazinon for fingerling European catfish were estimated as 14.597 (12.985-16.340), 12.487 (11.079-14.471), 8.932 (7.907-10.348), 6.326 (no data because of p > 0.05), and 4.142 (no data because of p > 0.05) mg L⁻¹, respectively. Compared to the control specimens, fish after an acute exposure to diazinon was significantly lower erythrocyte, leukocyte, hemoglobin, hematocrit, MCV, MCH, and MCHC values (p < 0.05). In addition, it was also showed a significantly negative correlation between these hematological parameters and exposure times of diazinon (p < 0.01).     

Fate of the herbicides glyphosate, glufosinate-ammonium, phenmedipham, ethofumesate and metamitron in two Finnish arable soils

The fate of five herbicides (glyphosate, glufosinate-ammonium, phenmedipham, ethofumesate and metamitron) was studied in two Finnish sugar beet fields for 26 months. Soil types were sandy loam and clay. Two different herbicide-tolerant sugar beet cultivars and three different herbicide application schedules were used. Meteorological data were collected throughout the study and soil properties were thoroughly analysed. An extensive data set of herbicide residue concentrations in soil was collected. Five different soil depths were sampled. The study was carried out using common Finnish agricultural practices and represents typical sugar beet cultivation conditions in Finland. The overall observed order of persistence was ethofumesate > glyphosate > phenmedipham > metamitron > glufosinate-ammonium. Only ethofumesate and glyphosate persisted until the subsequent spring. Seasonal variation in herbicide dissipation was very high and dissipation ceased almost completely during winter. During the 2 year experiment no indication of potential groundwater pollution risk was obtained, but herbicides may cause surface water pollution.     

A novel photodegradable insecticide: Preparation, characterization and properties evaluation of nano-Imidacloprid

Imidacloprid (IMI) microcrystals were directly encapsulated with nature polysaccharides chitosan (CHI) and sodium alginate (ALG) through layer-by-layer (LbL) self-assembly. The coated colloids were characterized using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). The in vitro controlled release pattern of IMI through the PE diffusion barrier was studied using a diffusion cell assembly at physiological pH of 7.4. Photocatalysts were characterized by Brunauer-Emmett-Teller (BET) surface area and SEM. The adsorption and photocatalytic activities of photocatalysts were evaluated by isothermal adsorption and IMI degradation under UV light and natural light illumination. The toxicity of the photodegradable insecticide was evaluated against the adult stage of Martianus dermestoides. The results showed that thermodynamically stable IMI microcrystals were obtained by association and had a mean length of 7 μm and a ζ-potential of −37.5. The drug loading and encapsulation efficiency were 56.15 ± 0.96% and 81.57 ± 0.96%, respectively. The polysaccharide capsules prolonged the release time of the encapsulated IMI crystals. Among the photocatalysts, SDS/Ag/TiO₂ had the highest photocatalytic activity. Toxicity of the novel 50% nano-SDS/Ag/TiO₂-IMI was higher in the adult stage compared to the 95% IMI as indicated by the lower LC₅₀ value.     

Biphasic responses of the honeybee heart to nanomolar concentrations of amitraz

Amitraz is a pesticide targeting the octopaminergic receptors. In a previous study, octopamine, a biogenic amine, was found to induce a biphasic effect on the honeybee heart, inhibition at low concentrations and excitation at high concentrations. Furthermore, the honeybee heart was found to be far more sensitive to octopamine compared to other insect hearts. The objective of the present study was to investigate the effects of amitraz on the electrical and mechanical properties of the honeybee heart ex vivo and on the heart rate in vivo. In ex vivo conditions, amitraz at 10⁻¹² M caused a significant inhibition in the mechanical (p < 0.05, n = 4) and electrical properties (p < 0.05, n = 4). Higher concentrations such as 10⁻⁹ and 10⁻⁶ M induced a biphasic effect, with total inhibition for 7.86 ± 1.26 min (n = 7), followed by strong excitation of spontaneously-generated contractions (n = 7). The initial elimination of heart activity was caused by strong hyperpolarization, while the subsequent excitation was caused by a depolarization in the membrane potential of pacemaker cells at 10⁻⁹ M (n = 8). In the in vivo experiments, abdominal injection or oral application of 0.20 ng of amitraz per bee induced a persistent increase of 134.28 ± 4.07% (p < 0.05, n = 4) in the frequency of the cardiac action potentials. The above responses clearly show that the heart of the honeybee is extremely vulnerable to amitraz, which is nevertheless still used inside beehives, ostensibly to “protect” the honeybees against their main parasite, Varroa destructor.     

Investigation of acute toxicity of (2,4-dichlorophenoxy)acetic acid (2,4-D) herbicide on crayfish (Astacus leptodactylus Esch. 1823)

The acute 96 h LC₅₀ of (2,4-dichlorophenoxy)acetic acid (2,4-D), a widely used agricultural herbicide, was determined on crayfish (Astacus leptodactylus Esch. 1823). Crayfish of 23.5 ± 1.49 g mean weight and 9.6 ± 0.21 cm mean length were selected for the bioassay experiments. The experiments were repeated three times, in 10 L tap water. The data obtained were statistically evaluated by the use of the E.P.A computer program based on Finney’s probit analysis method and the 96 h LC₅₀ value for crayfish was calculated to be 32.6 mg/L in a static bioassay test system. 95% lower and upper confidence limits for the LC₅₀ were 15.10-327.16. In conclusion, 2,4-D is highly toxic to crayfish, a non-target organism in the ecosystem. Water temperature was 23 ± 1 °C. Behavioral changes of crayfish were recorded for all herbicide concentrations.     

Effect of waterborne benomyl on the hematological and antioxidant parameters of the Nile tilapia, Oreochromis niloticus

The present study was conducted to determine the 96 h-LC₅₀ of benomyl to the Nile tilapia, Oreochromis niloticus and to investigate the biochemical or hematological indices of blood and the alterations in the antioxidant enzymes of this fish in response to sublethal concentrations of benomyl. Fish weighing 71.61 ± 12.05 g were used in this study; they were subjected to fasting for 4 weeks before treatment. An aqueous solution of benomyl (0, 0.5, 1, 2, 4, 8, and 16 mg L⁻¹) was administered for 96 h to determine the LC₅₀. The 96 h-LC₅₀ value of benomyl was 4.39 (3.23-5.60) mg L⁻¹ in the present study. For 5 weeks, the aqueous solution of benomyl (0, 100, 200, and 400 μg L⁻¹) was administered to investigate its effect on the hematological parameters and antioxidant enzymes. The predominant hematological findings in fish exposed to benomyl were as follows: no significant change in the Hb (g dL⁻¹) level, MCV (μm³), MCH (pg) and MCHC (%) as compared to the control. Benomyl exposure led to greater increases in the GPT, GOT (Karmen-unit), LDH (Wroblewski unit), total cholesterol, Fe, and Ca (mg dL⁻¹) values, whereas the levels of ALP (KA unit), total protein, triglyceride, albumin, and Mg (mg dL⁻¹) did not increase. Benomyl increased the in vivo HSI (%), GST (nmol min⁻¹ mg protein⁻¹), and SOD (U mg protein⁻¹) values in the fish livers in the test group, unlike those in the control group for 5 weeks. At concentrations higher than 100 μg L⁻¹, benomyl affected the GST and SOD levels of Nile tilapia in a dose- and time-dependent manner. The present findings suggest that the in vivo hepatotoxicity associated with benomyl may, in part, result from the hematological index, and antioxidants may provide limited protection against benomyl toxicity.     

Effects of gender and temperature on oxidative stress enzymes in Nile tilapia Oreochromis niloticus exposed to paraquat

Many classes of environmental pollutants can enhance the intracellular formation of reactive oxygen species, which can conduce to the damage of macromolecules and changes in oxidant defences levels in fish. In the present study it was analysed the hepatic levels of superoxide dismutase (SOD), glutathione reductase (GR), and glutathione S-transferase (GST) in males and females of Nile tilapia Oreochromis niloticus exposed to paraquat (PQ), at 17 and 27 °C. Tilapia were exposed to a sublethal concentration of PQ (0.5 mg L⁻¹) during 45 days. Condition factor and hepatosomatic index of males and females exposed to PQ were significantly higher when compared with the control group, except in females at 17 °C. SOD and GST activities were higher in males and females exposed to PQ than in the control group at 17 and 27 °C. The levels of both enzyme activities revealed that they are sex-dependent with males exposed to PQ showing higher SOD activity (5.05 ± 0.13 and 4.84 ± 0.23 U/g protein, respectively at 17 and 27 °C) than females (4.21 ± 0.07 and 3.87 ± 0.27 U/g protein, respectively at the same temperature). Similar results were observed in GST activity. A GR activity significantly higher (9.09 ± 0.44 and 7.97 ± 1.08 U/g protein at 17 and 27 °C, respectively) was observed in PQ-exposed females, but not in exposed males. Fish exposed to PQ showed higher values of SOD and GST activities than the control group at both temperatures. These results are gender-dependent, while GR activity was higher only in PQ-exposed females. No significant differences were found for SOD, GST and GR activities between fish exposed to 17 and 27 °C, although males and females showed higher values at 17 °C. In short, this work advanced new knowledge on influence of gender in same biochemical parameters in tilapia exposed to PQ and demonstrated that their effects could be observed at different temperatures.     

Stereoselective metabolism of fipronil in water hyacinth (Eichhornia crassipes)

The enantioselective metabolism of racemic fipronil in water hyacinth (Eichhornia crassipes) had been investigated. In this study, the degradation data and the enantiomer fraction (EF) were determined by chiral high-performance liquid chromatography (HPLC) with a column cellulose-tri-(3, 5-dimethylphe-nylcarbamate)-based chiral stationary phase (CDMPC-CSP). During the uptake phase, the EF value of plant sample increased from 0.50 at 1st day to 0.72 at 63rd day, while it was almost unchanged in water. For the depuration phase, the S- and R-enantiomer of fipronil in water hyacinth plants were degraded 92.22% and 82.07% after 17 days, respectively. The process of the degradation of the two enantiomers was followed first-order kinetics (R² ? 0.94). Stereoselective behavior was observed in both accumulation and degradation process. In this study, fipronil-sulfone and fipronil-sulfide, the metabolites of fipronil, were detected by GC-MS to show the main metabolic pathway of fipronil in water hyacinth.     

Comparison of the inhibition effect of different anticoagulants on vitamin K epoxide reductase activity from warfarin-susceptible and resistant rat

Anti-vitamin K drugs are widely used as anticoagulant in human thromboembolic diseases. Similar compounds have also been used as rodenticides to control rodent population since 1950s. Massive use of first generation anticoagulants, especially warfarin, has lead to the development of genetic resistances in rodents. Similar resistances have been reported in human. In both cases, polymorphisms in VKORC1 (Vitamin K epoxide reductase subunit 1), the subunit 1 of the VKOR (Vitamin K epoxide reductase) complex, were involved. In rats (Rattus norvegicus), the Y139F mutation confers a high degree of resistance to warfarin. Little is known about the in vitro consequences of Y139F mutation on inhibitory effect of different anticoagulants available. A warfarin-susceptible and a warfarin-resistant Y139F strain of wild rats (Rattus norvegicus) are maintained in enclosures of the Lyon College of Veterinary Medicine (France). Using liver microsomes from susceptible or resistant rats, we studied inhibition parameters by warfarin (K_i = 0.72 ± 0.1 μM; 29 ± 4.1 μM), chlorophacinone (K_i = 0.08 ± 0.01 μM; 1.6 ± 0.1 μM), diphacinone (K_i = 0.07 ± 0.01 μM; 5.0 ± 0.8 μM), coumachlor (K_i = 0.12 ± 0.02 μM; 1.9 ± 0.2 μM), coumatetralyl (K_i = 0.13 ± 0.02 μM; 3.1 ± 0.4 μM), difenacoum (K_i = 0.07 ± 0.01 μM; 0.26 ± 0.02 μM), bromadiolone (K_i = 0.13 ± 0.02 μM; 0.91 ± 0.07 μM), and brodifacoum (K_i = 0.04 ± 0.01 μM; 0.09 ± 0.01 μM) on VKOR activity. Analysis of the results leads us to highlight different anticoagulant structural elements, which influence inhibition parameters in both susceptible and Y139F resistant rats.     

Determination of carbendazim with multiwalled carbon nanotubes-polymeric methyl red film modified electrode

Multiwalled carbon nanotubes-polymeric methyl red film modified electrode (MWNT-PMRE) was made. The electrochemical behavior of carbendazim on modified electrode was studied with Cyclic Voltammetry, Linear Sweep Voltammetry, Stable Polarization Method and Chronocoulometry. The results indicated that the electrical oxidation of carbendazim on MWNT-PMRE in H₂SO₄ supporting electrolyte with concentration of 0.6 mol/L was irreversible and was mainly controlled by diffusion. Some parameters of the electrochemical process were evaluated. The impacts of experiment conditions on the electrochemical behavior of carbendazim were studied. A good linearity relationship between peak current and concentration of carbendazim in the range of 2.0 × 10⁻⁷-1.0 × 10⁻⁵ mol/L was found, of which the equation was I_p(A) = −1.149 × 10⁻⁵ − 2.301c (mol/L), the correlative coefficient R = −0.9953 and detection limit was 9.0 × 10⁻⁹ mol/L. The recovery was between 90.3% and 94.7%.