Some toxicological aspects op methamidophos exposure in mice 1

Abstract

Cholinesterase activity in the brain, RBC and plasma of Swiss mice was determined following different routes of administration of methamidophos. Continuous feeding with the insecticide caused a progressive inhibition of both plasma‐ and erythrocyte enzymes. The effect of methamidophos was more pronounced when applied in diet than when administered dermally or intraperitoneally. Following a single injection (i.p.) of methamidophos, the brain enzyme showed maximum inhibition 24 hr following treatment. At the appearance of tremors, the plasma and RBC‐enzymes showed considerable inhibition, the former being more inhibited. The plasma enzyme appears to be the most sensitive enzyme and may be taken as a suitable index for exposure to methamidophos.     

Toxicity and metabolism of acephate in adult and larval insects

Abstract

Adult and larval insects from the terrestrial and aquatic environments were exposed to acephate. The chemical was more toxic to adult insects than to larvae, and was a poor insect cholinesterase inhibitor in vitro compared to methamidophos which was a much stronger inhibitor. Both acephate and methamidophos inhibited the adult cholinesterase in vitro much more strongly than they did the larval enzymes. Acephate was metabolized by the insects to methamidophos which did not appear to be the only metabolite, although no other metabolites were looked for. The cholinesterase of insects exposed to sublethal levels of acephate was inhibited, but this inhibition appeared to be due to the combined effect of acephate and methamidophos and not to any hypothetical substance with greater anticholinesterase activity. This was bourne out when acephate was incubated with mixed function oxidases (MFO). No activated product with potent anticholinesterase activity was identified. Methamidophos was not produced by the MFO system but by some other unidentified mechanism.     

Hydrolytic and metabolic products of acephate in water and mouse liver

Acephate was incubated in distilled water of three different pH's at 37 degrees C for 7 days. Three hydrolytic products were formed: methamidophos, O,S-dimethyl phosphorothiolate (DMPT), and O-methylacetyl phosphoramidothiolate (OMPT). A single dose of acephate was also fed to mice, and their livers were excised and analyzed for metabolic products up to 30 hours. Three products were detected: methamidophos, DMPT, and S-methylacetyl phosphoramidothiolate (SMPT). The anticholinesterase properties of acephate, methamidophos, DMPT, SMPT, and OMPT were determined. Only acephate and methamidophos had measurable inhibitory effects on the mouse erythrocyte enzyme, methamidophos being about ten times more effective than acephate. The amount of methamidophos formed in the water and mouse liver was too low to have any direct effect on the toxicity of acephate. Acephate toxicity to aquatic insects would depend on its persistence in water, its uptake by the insects, its conversion to methamidophos, and the combined inhibitory effect of acephate and methamidophos on the cholinesterase enzyme. The toxicity of acephate to mammals would depend on the direct anticholinesterase effect of the chemical and to a small extent on methamidophos.     

几种农药对鲤鱼脑AchE的联合毒性效应

采用体内染毒的方法,以鲤鱼脑乙酰胆碱酯酶(AchE)活力为指标,研究了有机磷农药对硫磷与氯乐果、甲胺磷涕灭威之间的联合毒性效应.结果表明,这些农药之间均产生较强的协同作用,但是两种农药以不同比例加入,其产生的毒性效应有明显差别,涕灭威/对硫磷之间的协同作用要强于同类农药间的作用.     

Studies on the toxicity, metabolism, and anticholinesterase properties of acephate and methamidophos

The toxicity of acephate to four species of aquatic insects, as well as the metabolism and cholinesterase-inhibiting properties of the chemical in the rat were studied. The results indicated that mayfly larvae were very sensitive to the toxic effects of acephate, whereas larvae of the stonefly, damselfly and mosquito were much less sensitive. In the rat, orally-administered acephate was rapidly absorbed from the intestines and severely inhibited the cholinesterases in the blood and brain. The enzymes began to recover after 24 hours, while the chemical was completely eliminated within three days. The amount of methamidophos observed in the liver was extremely low. The cholinesterase-inhibiting properties of acephate and methamidophos were compared in vitro to that of paraoxon, a known strong anticholinesterase. Enzymes from four vertebrates were used. In all cases, except one, acephate was found to be six orders of magnitude weaker than paraoxon, whereas methamidophos was three orders weaker. Trout brain cholinesterase was the exception; it was as sensitive to paraoxon as it was to methamidophos. Finally, four cholinesterases were inhibited with methamidophos, and their ability to reactivate spontaneously or to recover by induction with pyridine aldoxime methiodide (PAM) in vitro were determined. The results suggested that methamidophos-inhibited cholinesterases did not reactivate spontaneously; instead the enzymes remained inhibited either in a phosphorylated or an aged state. The significance of these results are discussed in relation to the use of acephate for forest insect pests.     

Dimethoate-induced oxidative stress and DNA damage in Oncorhynchus mykiss

The present study was conducted in order to investigate pro-oxidant activity of dimethoate in liver and brain tissues following sublethal pesticide exposure for 5, 15 and 30 d by using SOD, GPx, CAT enzyme activities and lipid peroxidation as biomarkers as well as DNA damaging potential via detecting% Tail DNA, Tail moment and Olive tail moment as endpoints in erythrocytes of Oncorhynchus mykiss in an in vitro experiment. Antioxidant enzyme activities were found to elicit two staged response which was an initial induction followed by a sharp inhibition in liver tissue while a sustained increase in GPx activity and slight stimulation in SOD activity were detected in brain tissue. Lipid peroxidation showed an ascending pattern throughout the exposure period in both tissues and a decreasing trend was determined in tissue protein levels which was proved to be positively correlated with duration. Similar findings were obtained from outcomes preferred to quantify DNA damage and TM was decided to reflect the extent of damage more sensitively because of determined positive correlation with concentrations applied. Considering these results, it can be concluded that oxidative stress condition evoked by dimethoate could not be responded effectively and genotoxic nature of pesticide was proven by determined clastogenic effect possibly via being an alkylation agent or stimulating the production of reactive species.     

Importance of cholinesterase kinetic parameters in environmental monitoring using estuarine fish

The aim of the present study was to determine the kinetic parameters and cholinesterase (ChE) activity in two teleost fish, the white mouth croaker Micropogonias furnieri (Scianidae) and the Madamango sea catfish Cathorops spixii (Ariidae), to verify their suitability as sentinel species of aquatic pollution by anticholinergic compounds. Individuals of each species were captured in one reference and one polluted site in the Southern Brazilian coast. Brain tissue was used as enzyme source. Inhibition kinetic parameters indicated that ChE from C. spixii collected at the reference site showed more affinity (Ka) for eserine that from those collected at the polluted site. The opposite was observed for the carbamylation constants (Kc). Thus, similar inhibition constants (Ki) were observed. M. furnieri brain showed an extremely low sensitivity to in vitro inhibition by eserine, indicating that it is not a suitable biomarker to be employed in environmental monitoring of anticholinergic compounds. Results from the present study also point to the need for considering kinetic analysis when cholinesterase activity is employed as a biomarker.     

Analysis and fate of acephate and its metabolite,methamidophos, in pepper and cucumber

Abstract

Levels of acephate (Orthene^R) and its principle metabolite, methamidophos, in/on greenhouse‐grown pepper and cucumber fruits and leaves in relation to the applied methamidophos were monitored. Dislodgeable and total residues of acephate and methamidophos were determined by gas‐liquid chromatography equipped with a flame ionization detector (GC‐FID) and were confirmed by nitrogen phosphorus detector (GC‐NPD). The dissipation curves of the residues followed first‐order kinetics (R²> 0.96). Initial residues of acephate on fruits varied between pepper (15.12 ppm) and cucumber (2.16 ppm) . Total residues in fruits and leaves determined at intervals following application revealed the greater persistence of acephate on pepper fruits (half‐life [t_1/2] of 6 d) than on cucumber fruits (t_1/2 was 3.7 d) . T_1/2 values for the applied methamidophos were 4.7 and 5.3 d on pepper and cucumber fruits, respectively. Deacety‐lation of acephate (formation of its metabolite) was detectable 1 d following acephate treatment and reached a maximum of 2.05% of initial acephate residues 3 d after application on pepper fruits. On cucumber fruits, acephate metabolite reached a maximum of 2.12% one wk following application. No acephate residues were detected above the limit of detection of 0.001 ppm in pepper fruits 50 d following acephate application while its metabolite was detectable at that time (detectability limit was 0.0001 ppm).     

The fate of acephate and carbaryl in water.

Acephate was resistent to hydrolysis in distilled, buffered water at pH 4.0 to 6.9, but not at pH 8.2, held for 20 days at 20 or 30 degrees C. The maximum conversion to methamidophos was 4.5% of the added acephate at pH 8.2 and 20 degrees C. The persistence of acephate in two natural waters, held at 9 degrees C for up to 42 and 50 days varied: 80% were recovered from pond water after 42 days, and 45% from creek water after 50 days. Rates of acephate degradation increased greatly when treated water samples were incubated in the presence of sediments, but not if water and sediment were autoclaved prior to treatment and incubation. The greatest conversion to methamidophos, 1.3% of the added acephate, had occurred after 42 days in pond water without sediment. Under the same conditions, carbaryl was less persistent than acephate in the natural waters: 18 to 20% were recovered from pond water after 42 days, and 37 to 40% from creek water after 50 days. The presence of sediment did not affect its degradation significantly. But more than 55% were recovered after 50 days if water and sediment were autoclaved prior to treatment and incubation. Neither acephate, methamidophos, nor carbaryl could be shown to escape from water into the atmosphere.     

Adsorption,desorption, and mobility of two insecticides in Malaysian agricultural soil

Laboratory studies utilizing radioisotopic techniques were conducted to determine the adsorption, desorption, and mobility of endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxanthiepin3-oxide) and methamidophos (O,S-dimethyl phosphorothioate) in sandy loam and clay soils of the Cameron Highlands and the Muda rice-growing area, respectively. High Freundlich adsorption distribution coefficients [Kads(f)] for endosulfan (6.74 and 18.75) and low values for methamidophos (0.40 and 0.98) were obtained in the sandy loam and clay soils, respectively. The observed Koc values for endosulfan were 350.85 (sandy loam) and 1143.19 (clay) while Koc values of 20.92 (sandy loam) and 59.63 (clay) were obtained for methamidophos. Log Kow of 0.40 and 1.25 were calculated for endosulfan as well as -1.96 and -1.21 for methamidophos in the sandy loam and clay soils, respectively. Desorption was common to both pesticides but the desorption capacity of methamidophos from each soil type far exceeded that of endosulfan. Soil thin layer chromatography (TLC) and column studies showed that while methamidophos was very mobile in both soils, endosulfan displayed zero mobility in clay soil.     

Protective effects of isoflavone on some biochemical parameters affected by cypermethrin in male rabbits

Effect of isoflavone on cypermethrin-induced changes in enzyme activities and free radicals was studied in plasma, liver, brain and testes of male New Zealand White rabbits. Rabbits were orally given sublethal dose of cypermethrin (24 mg/kg BW; 1/100 LD50), while isoflavone (2 mg/kg BW) was given alone or in combination with cypermethrin. The tested doses were given to rabbits every other day for 12 weeks. Results obtained showed that cypermethrin significantly (P < 0.05) induced free radicals in plasma, liver, brain and testes. The activities of glutathione S-transferase (GST) (liver, brain and testes), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) (liver and testes), and alkaline phosphatase (AlP) (liver) were significantly (P < 0.05) decreased due to cypermethrin administration. Contrariwise, the activities of GST, AST, ALT and AIP were increased in plasma. The activity of acetylcholinesterase (AChE) did not change in plasma and brain of treated rabbits with cypermethrin. Isoflavone alone significantly (p < 0.05) decreased the levels of free radicals in plasma, liver, brain and testes, while did not produce any significant effect on the investigated enzymes. However, isoflavone is able to reverse the changes in enzyme activities due to the effect of cypermethrin. Results concluded that isoflavone confers marked protection against cypermethrin-induced oxidative stress in rabbit's plasma, liver, brain and testes.     

Studies of methamidophos-C-14 in Costa Rican vegetables and soils

In order to see the effect of time lapse between the last application of methamidophos and harvesting insecticide was applied on lettuce plants (6,84 μCi in one experiment and 4,03 μCi in the other experiment). Analysis of the crops harvested 3 days after last application showed 9,7 ppm residues on leaves, while crops harvested 1 day after application showed residues of 12,7 ppm (25% more). Treatment of tomato plants (39,65 μCi, 1,01 kg/ha) gave residues in fruits 4,92 ppm after 8 days interval between last application and harvesting. 40 days gap between the last application and harvesting leaved residues of 0,7 ppm in fruits which is much less as recommended by FAO/WHO (1 – 2 ppm).Degradation of this insecticide is dependent on the matrix of the soil, this breakdown is observed in the first ten days and than after it remains constant. C-14 radioactivity extracted from soil and plant analysis was methamidophos (92%)     

Degradation of disulfoton,oxydemeton‐methyl,methamidophos and demeton in asparagus plant

Abstract

Disulfoton and methamidophos (both at 1.12 kg a.i./ha), oxydemeton‐methyl and demeton, (both at 0.56 kg a.i./ha) were applied as post‐harvest foliar sprays to control the European asparagus aphid, Brachycolus asparagi. Oxidation of disulfoton, oxydemeton‐methyl and demeton to their corresponding sulfoxides and sulfones occurred in asparagus foliage 2 to 5 days after application. The total residues of these three compounds, including their toxic oxidative metabolites declined to less than 0.5 ppm about 47 days after the spray application whereas methamidophos persisted longer; 0.84 ppm of its residue was found even after 85 days. No residue was found above the limit of detection of 0.002 ppm in any asparagus spears which were produced in the following spring; the four compounds were sprayed on the asparagus plants during the previous season at realistic rates for aphid control.     

Effects of the aflatoxins on ATPase activities in mouse and rat liver.

The effects of the aflatoxins on ATPase activities in mouse and rat tissues were investigated in vitro. The hepatic oligomycin-sensitive (O.S.) Mg++ ATPase was inhibited significantly. The order of inhibition was G1 greater than B1 greater than G2 greater than B2. Mouse O.S. Mg++ ATPase was more sensitive than the corresponding rat enzyme. The oligomycin-insensitive (O.I.) Mg++ ATPase activities in rat and mouse liver were not altered. Although aflatoxins G1 and B1 were more potent inhibitors of hepatic O.S. Mg++ ATPase, no concentration-response was observed, whereas aflatoxins G2 and B2 inhibited enzyme activity in a concentration-dependent manner. Spectral analysis of aflatoxin G1 solutions suggested that solubility was not related to the observed effects. In addition, the effects of aflatoxin B1 and G1 on mouse brain microsomal Na+-K+ ATPase were examined. Although aflatoxin B1 was more potent that G1, both mycotoxins significantly inhibited enzyme activity in a concentration-dependent fashion.     

Survival and behavior of the insecticide-exposed predators Podisus nigrispinus and Supputius cincticeps (Heteroptera: Pentatomidae)

Pentatomid stinkbugs are important predators of defoliating caterpillars in agricultural and forestry systems, and knowledge of the impact of insecticides on natural enemies is important information for integrated pest management (IPM) programs. Thus, we assessed the toxicity and behavioral sublethal response of the predators Podisus nigrispinus and Supputius cincticeps exposed to deltamethrin, methamidophos, spinosad and chlorantraniliprole, insecticides commonly used to control the velvetbean caterpillar (Anticarsia gemmatalis) in soybean crops. With the exception of deltamethrin for S. cincticeps, all insecticides showed higher acute toxicity to the prey than to these natural enemies providing effective control of A. gemmatalis. The recommended field concentration of deltamethrin, methamidophos and spinosad for controlling A. gemmatalis caused 100% mortality of P. nigrispinus and S. cincticeps nymphs. Chlorantraniliprole was the less toxic and the most selective insecticide to these predators resulting in mortalities of less than 10% when exposed to 10× the recommended field concentration for a period of 72 h. Behavioral pattern changes in predators were found for all insecticides, especially methamidophos and spinosad, which exhibited irritability (i.e., avoidance after contact) to both predator species. However, insecticide repellence (i.e., avoidance without contact) was not observed in any of the insects tested. The lethal and sublethal effects of pesticides on natural enemies is of great importance for IPM, and our results indicate that substitution of pyrethroid and organophosphate insecticides at their field rates by chlorantraniliprole may be a key factor for the success of IPM programs of A. gemmatalis in soybeans.     

Effect of methamidophos and urea application on microbial communities in soils as determined by microbial biomass and community level physiological profiles

In this study, we evaluated the effect of the application by two agrochemicals, methamidophos (O,S-dimethyl phosphoroamidothioate) and urea, on microbial diversity in soil, using the combined approaches of soil microbial biomass analysis and community level physiological profiles (CLPPs). The results showed that both a low and a high level of methamidophos application (CS2 and CS3) and urea application (CS4) significantly decreased microbial biomass C (Cmic) by 41-83% compared with the control (CS1). The soil organic C (Corg) values of CS3 and CS4 were significantly higher and lower by 24% and 14%, respectively, than that of CS1. Similarly to Cmic, the values of Cmic/Corg of the three applied soils which decreased were lower by 31-84% than that of CS1. In contrast, the respiration activity of the three applied soils were significantly higher than the control. Agrochemical application also significantly increased the soil total of N and P (Ntol and Ptol) and decreased the Corg/Ntol and Corg/Ptol values. The CLPPs results showed that the AWCD (average well color development) of the three applied soils were significantly higher than that of CS1 during the incubation period. Substrate richness, Shannon and Simpson indices of microbial communities under chemical stresses, increased significantly. In addition, the CFU (colony-forming unit) numbers of methamidophos metabolized bacteria in CS2 and CS3 also increased significantly by 86.1% and 188.9% compared with that of CS1. The combined results suggest that agrochemicals reduce microbial biomass and enhance functional diversities of soil microbial communities; meanwhile, some species of bacteria may be enriched in soils under methamidophos stress.     

Suppression of aromatase activity in populations of bream (Abramis brama) from the river Elbe, Germany

Aromatase activity was determined in brain and gonads of wild bream collected along the river Elbe, Germany, and correlated with other endocrine and reproductive endpoints such as plasma sex steroid concentrations, secondary sex characteristics (STI), plasma vitellogenin, gonad size (GSI), and maturation stages of germ cells (MS) that were reported for the same fish in a previous study. Furthermore, regional patterns of aromatase activity were correlated to a number of environmental factors such as exposure to environmental contaminants and parasitism. While aromatase activity was not detectable in the gonads of male and female fish with the assay used, fish of both genders revealed relatively great brain enzyme activities. As for most of the endocrine and reproductive parameters, with the exception of plasma testosterone (T), aromatase activities were significantly less in fish from a river stretch characterized by elevated exposures to organic contaminants and metals. Brain aromatase activity was positively and significantly correlated with plasma estradiol (E2) and MS in females, and showed a similar trend with plasma 11-ketotestosterone (11KT) and STI in males. No comparable trend occurred for T. This decrease of the reproductively relevant hormones 11KT and E2 may be indicative of a disruption of the last step in sex hormone synthesis, a hypothesis that was supported for E2 by the strong (R2=0.78, p<0.05) linear regression between aromatase activity and E2 in female bream. It is also hypothesized that the effects on brain aromatase activity were likely to be related to the disruption of other reproductive parameters including sexual maturity and expression of secondary sex characteristics. Although a number of factors such as exposure to pollutants and prevalence of the tapeworm Ligula intestinalis correlated with the suppression of aromatase activity, the exact causes for the regional decrease in brain aromatase activity remain unclear due to inconsistencies of these correlations between sampling events or gender.     

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.     

Avoidance behaviour of Eisenia fetida to carbofuran, chlorpyrifos, mancozeb and metamidophos in natural soils from the highlands of Colombia

Earthworm avoidance behaviour test is an important screening tool in soil eco-toxicology. This test has been developed and validated under North American and European conditions. However, little research has been performed on the avoidance test in the tropics. This work demonstrates the potential suitability of the avoidance behaviour test as screening method in the highlands of Colombia using Eisenia fetida as the bio-indicator species on contaminated soils with carbofuran and chlorpyrifos. Though for the two active ingredients 100% avoidance was not reached, a curve with six meaningful concentrations is provided. No significant avoidance behaviour trend was found for mancozeb and methamidophos. Tests were conducted in the field yielded similar results to the tests carried out in the laboratory for chlorpyrifos and mancozeb. However, for the case of carbofuran and methamidophos, differences of more than double in avoidance were obtained. Divergence might be explained by soil and temperature conditions.     

甲胺磷农药的微生物降解

综述了甲胺磷降解菌的分布、类型及其实际应用，并介绍了微生物代谢甲胺磷的可能机制。