Assessment of methomyl-induced adrenal gland disruption in rat fetuses and pups: Potential protective effects of propolis supplementation.

The present study aimed to unravel the possible adverse effects of methomyl on the developing adrenal gland of rat fetuses and pups. Additionally, this study explored the potential improving effects of propolis against these possible hazards induced by methomyl exposure. To achieve that, pregnant rats were divided into four groups: control group, received 1 mL distilled water, propolis group, received 1 mL propolis at a dose of 300 mg/kg, methomyl group, received 1 mL methomyl at a dose of 2 mg/kg, and combined group, received 1 mL methomyl followed by 1 mL propolis, an hour later at the same previous doses. The results revealed that methomyl exposure, during pregnancy and lactation, induced many histological and ultrastructural changes, caused DNA damage and downregulated the expression of steroidogenic acute regulatory (StAR) and CYP11B2 genes in the adrenal glands of both rat fetuses and pups. Interestingly, propolis supplementation demonstrated a remarkable ability to mitigate these deleterious effects and restored the histology and ultrastructure architecture of the adrenal glands of both fetuses and pups, as well as decreased DNA damage and upregulated the expression of StAR and CYP11B2 genes in the adrenal gland of rat fetuses and pups. In conclusion, our study highlights the potential hazardous impact of methomyl exposure during pregnancy and lactation on the development of the adrenal gland in rat fetuses and pups, moreover, the study presents a new approach to alleviate these effects through propolis administration which could be used as a dietary supplement to mitigate the adverse effects of methomyl exposure.

Appraisal of sub-chronic exposure to lambada-cyhalothrin and/or methomyl on the behavior and hepato-renal functioning in Oreochromis niloticus: Supportive role of taurine-supplemented feed.

The existing study was designed to inspect the toxicological consequences of two pesticides; lambda-cyhalothrin (LCT) and methomyl (MTM) and their combination on Nile tilapia (Oreochromis niloticus) behaviors, oxidative stress, hepato-renal function indices and microarchitectural alterations. In addition, the efficiency of taurine (TUR) to rescue their toxicity was also considered. Juvenile O. niloticus were assigned into eight groups. The control and TUR groups were fed on a basal diet and TUR-enriched (10 g kg1) diet, respectively. The other groups were fed on a basal diet, and exposed to LCT (0.079 µg L-1), MTM (20.39 µg L-1 and (LCT + MTM). The last three groups were (LCT + TUR), (MTM + TUR), and (LCT + MTM + TUR) and fed on a TUR-enriched diet during exposure to LCT and/or MTM for 60 days. The exposure to LCT and/or MTM resulted in several behavioral alterations and stress via enhanced cortisol and nor-epinephrine levels. A significant elevation of serum 8-hydroxy-2- deoxyguanosine, aspartate and alanine aminotransferases, lactate dehydrogenase, Alkaline phosphatase, urea, creatinine was also observed in these groups. Furthermore, reduced antioxidant enzymes activities, including (catlase, glutathione peroxidase, and superoxide dismutase) with marked histopathological lesions in both liver and kidney tissues were detected. The up-regulated Bax and down-regulated Bcl-2 proteins were expressed in the liver and kidney tissues of LCT and/or MTM -exposed groups. Interestingly, all the observed alterations in behaviors, biochemical indices, and histo-architecture of renal and hepatic tissues were mitigated by TUR supplementation. The findings suggest that feeding O. niloticus dietary TUR may help to reduce the negative effects of LCT and/or MTM, and can also support kidney and liver health in O. niloticus, making it a promising aquaculture feed supplement.

Signal optimized rough silver nanoparticle for rapid SERS sensing of pesticide residues in tea.

Trace detection of toxic chemicals in foodstuffs is of great concern in recent years. Surface-enhanced Raman scattering (SERS) has drawn significant attention in the monitoring of food safety due to its high sensitivity. This study synthesized signal optimized flower-like silver nanoparticle-(AgNP) with EF at 25 °C of 1.39 × 106 to extend the SERS application for pesticide sensing in foodstuffs. The synthesized AgNP was deployed as SERS based sensing platform to detect methomyl, acetamiprid-(AC) and 2,4-dichlorophenoxyacetic acid-(2,4-D) residue levels in green tea via solid-phase extraction. A linear correlation was twigged between the SERS signal and the concentration for methomyl, AC and 2,4-D with regression coefficient of 0.9974, 0.9956 and 0.9982 and limit of detection of 5.58 × 10-4, 1.88 × 10-4 and 4.72 × 10-3 µg/mL, respectively; the RSD value < 5% was recorded for accuracy and precision analysis suggesting that proposed method could be deployed for the monitoring of methomyl, AC and 2,4-D residue levels in green tea.

Multiple insecticide resistance in onion thrips populations from Western USA.

Thrips tabaci is a key pest of onions, especially in the Pacific Northwestern USA. Management of T. tabaci is dominated by the application of various insecticides. However, T. tabaci is known to develop insecticide resistance which possibly leads to control failures, crop loss, and environmental concern. Here, we evaluated resistance status of T. tabaci populations from conventional and organic commercial onion fields to three widely used insecticides: oxamyl, methomyl, and abamectin with on-field concentration-mortality bioassays. The biochemistry and molecular mechanisms underlying resistance to these insecticides were also investigated by using enzymatic assays and detecting resistance-associated mutations. Field-evolved resistance to oxamyl, methomyl and abamectin were detected in most of the T. tabaci populations collected from conventional onion farms. At the labeled field rate, all the tested insecticides, particularly methomyl and oxamyl, had significantly reduced efficacy. Enzymatic assays of insecticide target and detoxification enzymes indicated that T. tabaci populations in Western USA onions harbor multiple mechanisms of resistance including enhanced activities of detoxification enzymes and target site insensitivity. Our results provide new information in understanding the dynamics of T. tabaci adaptation to multiple insecticides, which will help to design sustainable insecticide resistance management strategies for T. tabaci. Furthermore, this study provides the foundation for future research in identifying the biochemical and molecular markers associated with insecticide resistance in T. tabaci.

Insecticide toxicity to the borer Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae): developmental and egg-laying effects.

Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae) is one of the major pests of solanaceous plants in South America. It is considered a great threat by the European and Mediterranean Plant Protection Organization due to the serious economic damage that it causes on tomato farms; therefore, controlling this pest is a challenging task in South America. Controlling N. elegantalis at the egg stage is the best way to prevent it from damaging crops; however, thorough studies about the effectiveness of chemicals on the different life stages of this insect pest are lacking. In this study, the effects of different chemical classes were evaluated on N. elegantalis adults, female oviposition behavior, larvae, eggs, and embryonic development. None of the tested insecticides demonstrated toxicity to the adults; however, the results showed that cartap hydrochloride affects oviposition behavior. Moreover, methomyl and cartap hydrochloride exhibited high toxicity against the eggs and larvae, with higher than 80% of mortality. These insecticides interrupted larval hatching and caused alterations in the chorion layer. Flubendiamide and deltamethrin demonstrated toxicity on N. elegantalis larvae; however, lufenuron, indoxacarb, methoxyfenozide, and chlorantraniliprole demonstrated low toxicity on both eggs and larvae, with lower than 70% of mortality. Fruit treated with cartap hydrochloride had a deterrent effect. The ovicidal activity revealed by methomyl and cartap hydrochloride might provide new approaches regarding insecticide effects on eggs. Methomyl, cartap hydrochloride, flubendiamide, and deltamethrin demonstrated toxicity on larvae. The evaluation of the chorion of the eggshell in this study has clarified the toxic effect of methomyl and cartap hydrochloride on eggs.

Protective effects of vitamin C and selenium supplementation on methomyl-induced tissue oxidative stress in adult rats.

Methomyl (MET) is used worldwide in agriculture and health programs. Besides its advantages in the agriculture, it causes several toxic effects. The objective of this study was to examine the potential ability of vitamin C and/or selenium (Se), to alleviate the oxidative damage parameters, against MET-induced changes in blood biochemical markers and oxidative damage in liver and kidney of male Wistar rats. The animals were randomized into five groups of eight each: group I served as control rats; group II received MET (8 mg/kg body weight (BW)) in drinking water; group III received both MET and vitamin C (200 mg/kg BW; by intraperitoneal injection); group IV received both MET and Se (0.6 mg/100 g BW). Animals of group V were treated with MET, vitamin C and Se. A significant increase in the levels of hepatic markers enzymes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase) was determined. Furthermore, renal markers such as urea and creatinine were increased in MET-treated rats. Additionally, serum cholesterol and triglycerides were significantly enhanced. Exposure of rats to MET caused significant increase in malondialdehyde levels, thus causing a drastic alteration in antioxidant defense system, particularly in the activities of catalase and glutathione-S-transferase and glutathione peroxidase. However, simultaneous supplementation with vitamin C and Se restored these parameters partially. In conclusion, the results of the current study revealed that MET-induced toxicity caused perturbations of some biochemical parameters, lipid peroxidation and alterations in the antioxidant enzymes in liver and kidney homogenates. Administration of vitamin C and Se exhibited protective effect by inhibiting MET-induced toxicity in liver and kidney.

Molluscicidal activity of methomyl and cardenolide extracts from Calotropis procera and Adenium arabicum against the land snail Monacha cantiana.

In this work, we have evaluated the molluscicidal activity of two cardenolide extracts from Adenium arabicum Balf f. [the benzene (B) and methanol (M) extracts], one cardenolide extract from Calotropis procera (Aiton) W.T. Aiton (extract C), and methomyl against the harmful land snail Monacha cantiana (Montagu). The contact LD50 values for the above mentioned plant extracts were 12.62, 34.63, and 34.35 mg·kg⁻¹ of body weight, respectively, while the LD50 for methomyl was 116.62 mg·kg⁻¹, that is, the plant extracts were 9.24, 3.37, and 3.4 times more toxic than methomyl. In addition, a simple colorimetric method, based on Kedde reagent, was modified to determine cardenolide concentrations in plant extracts. Thin layer chromatography analysis (TLC) showed several cardiac glycosidal compounds in each plant extract. The results proved that cardiac glycosides are promising candidate compounds that could be used to control land snails, or exploited to develop new, effective, and environmentally friendly molluscicides.

Rapid determination of carbamate pesticides in food using dual counter-current chromatography directly interfaced with mass spectrometry.

Dual counter-current chromatography (dual CCC)-tandem mass spectrometry (MS/MS) was successfully performed with a newly designed spiral column for dual CCC. The small column capacity required for directly coupling with electrospray MS/MS was accomplished by forming a rectangular spiral groove on a plastic disk and sealing it with a PTFE sheet. This novel dual CCC-MS/MS technique was successfully applied for the rapid determination of methomyl, fenobucarb and carbaryl pesticides in food. A two-phase solvent system of n-hexane-acetonitrile-0.1% formic acid (45:45:10) was suitable for both good dual CCC separation and sufficient ionization of pesticides. Recoveries of these three pesticides from mandarin orange and spinach samples fortified at 0.05 mg/kg were in the range of 93-107% with relative standard deviations of 2.4-3.8%.

Patterns of insecticide resistance in onion thrips (Thysanoptera: Thripidae) in onion fields in New York.

To develop an insecticide resistance management program for onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), on onions (Allium spp.), we surveyed populations in commercial onion fields in New York and evaluated their susceptibility to the two most widely used classes of insecticides plus two new insecticides during 2003-2005. All insecticide evaluations were conducted using the Thrips Insecticide Bioassay System (TIBS). As in our surveys conducted during 2002-2003, there were large temporal and spatial variations in susceptibility to the pyrethroid lambda-cyhalothrin (Warrior) across onion-growing regions in 2003. New data indicate that the field rate of methomyl (Lannate LV) still provides control but that the genes for resistance to methomyl are present in some populations. Tests with the two new insecticides, acetamiprid (Assail 70 WP) and spinosad (SpinTor 2CS), indicated they provided > 85% mortality at the field rate. To determine the spatial variation in insecticide susceptibility within a region, a series of systematic assays were conducted with lambda-cyhalothrin and methomyl. In 2004 and 2005, our data indicated that the within-region spatial variation in susceptibility to lambda-cyhalothrin was not large at the field rate or for the 100 ppm rate of methomyl. In 2005, a year in which T. tabaci densities in most fields were much higher than in 2004, growers were unable to control T. tabaci in particular fields and attributed this lack of control to resistance. Yet, we found similar levels of high susceptibility in all fields when using TIBS. This finding suggests that resistance had not developed and that variation in control may have been due to other factors, such as localized higher populations, poor spray coverage, too much time between spray applications, or different onion varieties.

Application of dual counter-current chromatography for rapid sample preparation of N-methylcarbamate pesticides in vegetable oil and citrus fruit.

Dual counter-current chromatography (dual CCC) has been successfully applied to rapid sample preparation for the simultaneous determination of residual carbaryl, fenobucarb and methomyl in vegetable oil and citrus fruit. The citrus fruit samples were extracted with n-hexane solution containing stable isotopically labeled internal standards (methomyl-d3, fenobucarb-d3 and carbaryl-d9), and applied to dual CCC using a two-phase solvent system of n-hexane-acetonitrile to purify the carbamate pesticides from aliphatic sample matrix. The coiled column was rotated at 420 rpm, the lower mobile phase was introduced through the head toward the tail, and the upper mobile phase in the opposite direction. Due to the high partition efficiency of dual CCC, the lower phase fraction collected from 2 to 5 min after injection could be subjected to flow-injection tandem mass spectrometry directly after concentration. Repetitive sample injection can be performed at high reproducibility without a risk of contamination from the compounds retained in the column.

Role of selenium on antioxidant capacity in methomyl-treated mice

Methomyl carbamate is a pesticide widely used in the control of insects. The presentwork aims at studying the effect of selenium on the antioxidant system ofmethomyl-treated mice. Swiss albino mice were intraperitoneally administered a singledose of methomyl (7 mg/Kg body weight). Mice of another group were injectedwith sodium selenite (5 µmole/Kg b.wt.) 7 days before methomyl intoxication. After24 hours, methomyl exposure resulted in significant increase in lactic dehydrogenaseactivity (LDH). The antioxidant capacity of hepatic cells in terms of the activities ofsuperoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST) and glutathione (GSH) content was diminished. Itappears that methomyl exerts its toxic effect via peroxidative damage to hepatic, renaland splenic cell membranes. Also, methomyl induced DNA damage in these organsas detected by alkaline filter elution technique. The distribution of methomyl in differentorgans of mice was detected by HPLC. Selenium administration prior tomethomyl injection produced pronounced protective action against methomyleffects. It is observed that selenium enhances the endogenous antioxidant capacity ofthe cells by increasing the activities of SOD, CAT, GR and GST as well as increasingGSH content. The activity of LDH was decreased in blood and the damage ofDNA was suppressed comparable to controls. In conclusion, the adverse effects ofmethomyl in mice could be ameliorated by selenium

A case of poisoning in a man who drank a nutrition supplement containing methomyl, a carbamate pesticide.

A 50-year-old man was admitted to the emergency room complaining oppression on his chest, sweating and vomiting. He had drunk a 30 ml volume nutrition supplement 60 minutes before. As myosis and decrease of serum choline esterase activity were observed on admission examination, poisoning was suspected and toxicological analyses were carried out on the heeltap of the drink. Drug screening by gas chromatography-mass spectrometry (GC/MS) revealed the presence of methomyl and the concentration of methomyl in the heeltap determined by liquid chromatography was 2.1 mg/ml. Methomyl concentrations in the serum and urine were determined after converting methomyl to its oxime form followed by derivatization and GC/MS. Methomyl concentration in the serum collected 6 hours after ingestion was 0.63 microg/ml, and that in the urine collected 7-20 hours after ingestion was 0.10 microg/ml. Based on these values and reported data, the amount of methomyl contaminated to the drink was considered to be a toxic dose.

Role of selenium on antioxidant capacity in methomyl-treated mice.

Methomyl carbamate is a pesticide widely used in the control of insects. The present work aims at studying the effect of selenium on the antioxidant system of methomyl-treated mice. Swiss albino mice were intraperitoneally administered a single dose of methomyl (7 mg/Kg body weight). Mice of another group were injected with sodium selenite (5 pmole/Kg b.wt.) 7 days before methomyl intoxication. After 24 hours, methomyl exposure resulted in significant increase in lactic dehydrogenase activity (LDH). The antioxidant capacity of hepatic cells in terms of the activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST) and glutathione (GSH) content was diminished. It appears that methomyl exerts its toxic effect via peroxidative damage to hepatic, renal and splenic cell membranes. Also, methomyl induced DNA damage in these organs as detected by alkaline filter elution technique. The distribution of methomyl in different organs of mice was detected by HPLC. Selenium administration prior to methomyl injection produced pronounced protective action against methomyl effects. It is observed that selenium enhances the endogenous antioxidant capacity of the cells by increasing the activities of SOD, CAT, GR and GST as well as increasing GSH content. The activity of LDH was decreased in blood and the damage of DNA was suppressed comparable to controls. In conclusion, the adverse effects of methomyl in mice could be ameliorated by selenium.

Determination and degradation of methomyl in tomatoes and green beans grown in greenhouses.

A liquid chromatographic (LC) method using UV detection at 233 nm was used to study the degradation of methomyl in tomatoes and green beans grown in greenhouses. A liquid-liquid extraction with CH2Cl2-methanol (90 + 10, v/v) and a cleanup step with Florisil were combined with LC to isolate, recover, and quantitate the pesticide. Average recoveries obtained at spike levels of 0.03 and 0.40 mg/kg were 83.2-84.7% for tomatoes and 83.3-87.5% for green beans. Determination limits were 0.03 mg/kg for tomatoes and 0.01 mg/kg for green beans. Levels of methomyl residues were studied in tomatoes and green beans grown in an experimental greenhouse to establish the effect of the kind of greenhouse, application dose, species grown, and climatic conditions on the degradation of this pesticide. Analysis of variance showed that doses did not affect the response. The half-life, however, is greater in a flat-roof greenhouse than in an asymmetric-roof greenhouse and is significantly longer for green beans than for tomatoes and longer in winter than in spring. A preharvest time of about 5 days may be suitable for green beans sprayed with methomyl. Tomatoes show residue levels at the time of application lower than Spanish minimum residue levels.

Targeting the maize T-urf13 product into tobacco mitochondria confers methomyl sensitivity to mitochondrial respiration.

The URF13 protein, which is encoded by the maize mitochondrial T-urf13 gene, is thought to be responsible for pathotoxin and methomyl sensitivity and male sterility. We have investigated whether T-urf13 confers toxin sensitivity and male sterility when expressed in another plant species. The coding sequence of T-urf13 was fused to a mitochondrial targeting presequence, placed under the control of the cauliflower mosaic virus 35S promoter, and introduced into tobacco by Agrobacterium tumefaciens-mediated transformation. Plants expressing high levels of URF13 were methomyl sensitive. Subcellular analysis indicated that URF13 is mainly associated with the mitochondria. Adding methomyl to isolated mitochondria stimulated NADH-linked respiration and uncoupled oxidative phosphorylation, indicating that URF13 was imported into the mitochondria, and conferred toxin sensitivity. Most control plants, which expressed the T-urf13c construct lacking the mitochondrial presequence, were methomyl sensitive and contained URF13 in a membrane fraction. Subcellular fractionation by sucrose gradient centrifugation showed that URF13 sedimented at several positions, suggesting the protein is associated with various organelles, including mitochondria. No methomyl effect was observed in isolated mitochondria, however, indicating that URF13 was not imported and did not confer toxin sensitivity to the mitochondria. Thus, URF13 confers toxin sensitivity to transgenic tobacco with or without import into the mitochondria. There was no correlation between the expression of URF13 and male sterility, suggesting either that URF13 does not cause male sterility in transgenic tobacco or that URF13 is not expressed in sufficient amounts in the appropriate anther cells.

[Thin layer chromatographic separation and enzymatic detection of carbamate pesticides in plant extracts]. / Dünnschichtchromatographische Trennung und enzymatischer Nachweis von Carbamatpestiziden in Pflanzenextrakten

The authors developed a thin-layer chromatographic technique for the separation of carbamate pesticides from plant material and an enzymatic method for their determination. The cellulose plates were impregnated with polar substances (ethylene glycol, triethylene glycol and beta,beta'-oxydipropionitrile). n-Heptane/chloroform and iso-octane/dioxane served as solvents. The slight enzyme-inhibiting effect and the colouration of the plant extracts render the detection of the pesticides difficult. Nevertheless, the separation and the enzymatic detection of Methomyl, Carbaryl, Baygon and Aldicarb could be achieved. The Rf values of the carbamates are not influenced by the kind (apples, pears, onions) and the quantity (50--200 mg) of the plant material.