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.

Effect of environmental level of methomyl on hatching, morphology, immunity and development related genes expression in zebrafish (Danio rerio) embryo.

The extensive use of carbamate pesticides has led to a range of environmental and health problems, such as surface and groundwater contamination, and endocrine disorders in organisms. In this study, we focused on examining the effects of toxic exposure to the carbamate pesticide methomyl on the hatching, morphology, immunity and developmental gene expression levels in zebrafish embryos. Four concentrations of methomyl (0, 2, 20, and 200 µg/L) were administered to zebrafish embryos for a period of 96 h. The study found that exposure to methomyl accelerated the hatching process of zebrafish embryos, with the strongest effect recorded at the concentration of 2 µg/L. Methomyl exposure also trigged significantly reductions in heart rate and caused abnormalities in larvae morphology, and it also stimulated the synthesis and release of several inflammatory factors such as IL-1ß, IL-6, TNF-α and INF-α, lowered the IgM contents, ultimately enhancing inflammatory response and interfering with immune function. All of these showed the significant effects on exposure time, concentration and their interaction (Time × Concentration). Furthermore, the body length of zebrafish exposed to methomyl for 96 h was significantly shorter, particularly at higher concentrations (200 µg/L). Methomyl also affected the expression levels of genes associated with development (down-regulated igf1, bmp2b, vasa, dazl and piwi genes), demonstrating strong developmental toxicity and disruption of the endocrine system, with the most observed at the concentration of 200 µg/L and 96 h exposure to methomyl. The results of this study provide valuable reference information on the potential damage of methomyl concentrations in the environment on fish embryo development, while also supplementing present research on the immunotoxicity of methomyl.

Toxicity of methomyl insecticides to testicular cells and protective effect of folic acid.

Methomyl is a carbamate insecticide with confirmed testicular toxicity. This study intended to observe the effect of methomyl on testicular cells and the protective effect of folic acid through in vitro experiments. The GC-1 spermatogonia, TM4 Sertoli cells, and TM3 Leydig cells were treated with methomyl (0, 250, 500, and 1000 µM) with or without folic acid (0, 10, 100, and 1000 nM) for 24 h. It was found that methomyl increased cytotoxicity to testicular cells in a dose-dependent manner. In spermatogonia, methomyl significantly inhibited the expression of proliferation genes Ki67 and PCNA at 1000 µM, and increased the expression of apoptosis genes Caspase3 and Bax at each dose. In Sertoli cells, methomyl dose-dependently inhibited the expression of blood-testis barrier function genes TJP1, Cx43, and N-cadherin, but did not affect Occludin and E-cadherin. In Leydig cells, methomyl inhibited the expression of steroid synthase P450scc, StAR, Hsd3b1 and down-regulated the level of testosterone, but did not affect Cyp17a1 and Hsd17b1. Further, folic acid could basically reduce the damage caused by methomyl. This study provided new insights into the toxicity of methomyl and the protective effect of folic acid.

Effect of methomyl on water quality, growth performance and antioxidant system in liver of GIFT (Oreochromis niloticus) in the presence of peppermint (Mentha haplocalyx Briq.) as a floating bed.

This study was conducted to investigate the effect of methomyl (MET) on water quality, growth and antioxidant system of genetically improved farmed tilapia (GIFT, Oreochromis niloticus) in the presence of peppermint as a floating bed. The concentration of NH3-N, NO2--N, NO3--N and TP in T3 (with 200 g wet peppermint) was significantly lower (P < 0.05) than that in T2 (100 g), T1 (50 g) and control, and the nutrient removal rates were 61.90%, 31.59%, 59.86% and 45.92% in 20 days, respectively. Juveniles GIFT (5.1 ± 0.2 g) were exposed to sub-lethal concentrations of 0.2, 2.0, 20 and 200 µg/L of MET for 45 days. After 6 weeks of a feeding trial, percentage weight gain (PWG), specific growth rate (SGR) and feed conversion ratio (FCR) were significantly decreased in 0.2, 2.0, 20 µg/L MET groups respectively and increased in the 200 µg/L MET group. Compared with the control, no significant changes in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) were detected in the 0.2 µg/L group. The significant increase in activities of SOD, CAT and GPx was accompanied by a diminution in reduced glutathione (GSH) levels resulting with tilapia exposed to 2.0, 20, or 200 µg/L for 45 days. The highest rates observed in SOD, CAT, GPx were 157.63%, 164.05% and 167.46% of the control respectively, and the lowest inhibition rate in GSH was 66.42% of the control. Peppermint as a floating bed can alleviate the adverse effects of MET, such as growth retardation and oxidative stress.

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.

Neurotoxicity of singular and combined exposure of Oreochromis niloticus to methomyl and copper sulphate at environmentally relevant levels: Assessment of neurotransmitters, neural stress, oxidative injury and histopathological changes.

Aquatic organisms are concomitantly exposed to multiple noxious chemicals that can be discharged into water bodies. We aimed to investigate the single and simultaneous sub-acute exposure to copper and methomyl on juvenile Oreochromis niloticus. Compared to the controls, the outcomes revealed that brain of methomyl-exposed fish displayed significant declines in the activities of SOD, CAT, and GST in addition to higher MDA and lower GSH levels. Methomyl induced notable declines in levels of GABA and acetylcholine esterase in brain and muscle of exposed fish. Noteworthy downregulated gene expression levels of TNF-α, HSP-70 together with upregulated c-fos were evident in brain of fish expose to either of tested compounds. Marked apoptotic changes were observed in fish brain exposed to copper and methomyl indicated by augmented immune expression of caspase-3. Conclusively, the results indicated the possible interaction between both compounds with subsequent toxic effects that differ from their single exposure.

Zebrafish Larvae Rapidly Recover from Locomotor Effects and Neuromuscular Alterations Induced by Cholinergic Insecticides.

Owing to the importance of acetylcholine as a neurotransmitter, many insecticides target the cholinergic system. Across phyla, cholinergic signaling is essential for many neuro-developmental processes including axonal pathfinding and synaptogenesis. Consequently, early-life exposure to such insecticides can disturb these processes, resulting in an impaired nervous system. One test frequently used to assess developmental neurotoxicity is the zebrafish light-dark transition test, which measures larval locomotion as a response to light changes. However, it is only poorly understood which structural alterations cause insecticide-induced locomotion defects and how persistent these alterations are. Therefore, this study aimed to link locomotion defects with effects on neuromuscular structures, including motorneurons, synapses, and muscles, and to investigate the longevity of the effects. The cholinergic insecticides diazinon and dimethoate (organophosphates), methomyl and pirimicarb (carbamates), and imidacloprid and thiacloprid (neonicotinoids) were used to induce hypoactivity. Our analyses revealed that some insecticides did not alter any of the structures assessed, while others affected axon branching (methomyl, imidacloprid) or muscle integrity (methomyl, thiacloprid). The majority of effects, even structural, were reversible within 24 to 72 h. Overall, we find that both neurodevelopmental and non-neurodevelopmental effects of different longevity can account for the reduced locomotion. These findings provide unprecedented insights into the underpinnings of insecticide-induced hypoactivity.

Monitoring of Spotted-Wing Drosophila (Diptera: Drosophilidae) Resistance Status Using a RAPID Method for Assessing Insecticide Sensitivity Across the United States.

Drosophila suzukii (Matsumura) has spread rapidly, challenging berry and cherry crop production due to its ability to lay eggs into ripening fruit. To prevent infestation by this pest, insecticides are applied during fruit ripening and harvest. We field-tested the Rapid Assessment Protocol for IDentification of resistance in D. suzukii (RAPID) on seventy-eight populations collected across eight U.S. states in 2017 and 2018. Exposure to LC50 rates of malathion, methomyl, spinetoram, spinosad, and zeta-cypermethrin led to average female fly mortality of 25.0% in 2017, and after adjusting concentrations the average was 39.9% in 2018. Using LC99 × 2 discriminating concentrations in 2017 and LC90 × 8 rates in 2018, average female mortalities were 93.3% and 98.5%, respectively, indicating high overall susceptibility. However, using these high concentrations we found 32.0% of assays with survival of some female flies in 2017 and 27.8% in 2018. The adjustment in discriminating dose from 2017 to 2018 also reduced the proportion of assays with <90% survival from 17.6 to 2.9%. Populations with low mortality when exposed to spinosad were identified using this assay, triggering more detailed follow-up bioassays that identified resistant populations collected in California coastal region berry crops. Widespread evaluations of this method and subsequent validation in California, Michigan, and Georgia in 2019-2021 show that it provides a quick and low-cost method to identify populations of D. suzukii that warrant more detailed testing. Our results also provide evidence that important insecticide classes remain effective in most U.S. regions of fruit production.

Side effects of insecticides applied to cotton on adult Trichogramma pretiosum by three exposure routes.

BACKGROUND: Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) is released in extensive areas cultivated with cotton worldwide, but the use of synthetic insecticides threatens the establishment of augmented populations. Thus, an assessment of insecticides' effects on T. pretiosum is required to establish their compatibility with mass releases of the parasitoid. We studied in the laboratory the impact of insecticides administered through different exposure routes (direct-spraying, ingestion and residue contact) on T. pretiosum. Based on their toxicity, the insecticides were rated into four categories (harmless, slightly harmful, moderately harmful and harmful) as per the International Organization for Biological Control. RESULTS: The survival, parasitism ability and emergence of the treated adults and offspring (F1 and F2 ), in addition to the persistence of the toxic effects in semi-field conditions, were assessed. Teflubenzuron did not reduce female survival and caused a lesser impact on T. pretiosum on both direct-spraying and ingestion bioassays. In the residue contact bioassay, teflubenzuron and flupyradifurone were rated as harmless and slightly harmful, respectively. The other active substances (chlorfenapyr, thiodicarb and methomyl) were harmful to the parasitoid by all exposure routes and were persistent (with toxicity duration surpassing 30 days). CONCLUSION: From these findings, teflubenzuron is the insecticide most compatible with T. pretiosum releases and should be preferred over the other active substances. Further studies with the harmful insecticides (thiodicarb, chlorfenapyr and methomyl) are required to determine their toxicity under field conditions and confirm their incompatibility with T. pretiosum releases.

Subacute and subchronic methomyl exposure induced toxic effects on intestines via oxidative stress in male albino rats: biochemical and histopathological study.

The aim of the present study was to estimate the potential toxic effects of a subacute and a subchronic exposure to methomyl (MET) on some parts of the digestive system in male albino rats. Wistar rats were exposed daily by oral gavage to 4 mg/kg body weight (b.w.) of MET over 28 days in subacute experiments and to 1 mg/kg b.w. of MET for 90 consecutive days in subchronic exposure. Plasma levels of liver function indicators (ALT, AST, ALP and LDH) were significantly elevated after administration of MET to rats. MET had significant pro-oxidative effects on the intestines, as indicated by decreases in nonenzymatic (GSH) and enzymatic antioxidant (GPx, GST and CAT) parameters and significant increases in TBARS levels in both the duodenum and colon of rats. Microscopically, the duodenum and colon of rats exposed to MET showed severe pathological changes. It could be concluded that MET profoundly altered the structure and antioxidant status of intestinal rat tissue, which might lead to intestinal and digestive dysfunctions.

Toxic effects of methomyl on mouse oocytes and its possible mechanisms.

Methomyl is a broad-spectrum carbamate insecticide that has a variety of toxic effects on humans and animals. However, there have been no studies on the toxicity of methomyl in female mammalian oocytes. This study investigated the toxic effects of environmental oestrogen methomyl exposure on mouse oocyte maturation and its possible mechanisms. Our results indicated that methomyl exposure inhibited polar body extrusion in mouse oocytes. Compared with that in the control group, in the methomyl treatment group, superoxide anion free radicals in oocytes were significantly increased. In addition, the mitochondrial membrane potential of metaphase II stage oocytes in the methomyl treatment group was significantly decreased, resulting in reduced mouse oocyte quality. After 8.5 h of exposure to methomyl, metaphase I stage mouse oocytes displayed an abnormal spindle morphology. mRNA expression of the pro-apoptotic genes Bax and Caspase-3 in methomyl-treated oocytes increased, which confirmed the apoptosis. Collectively, our results indicated that mouse oocyte maturation is defective after methomyl treatment at least through disruption of spindle morphology, mitochondrial function and by induction of oxidative stress.

Investigation of pesticides on honey bee carbonic anhydrase inhibition.

Carbonic anhydrase (CA, EC 4.2.1.1) plays crucial physiological roles in many different organisms, such as in pH regulation, ion transport, and metabolic processes. CA was isolated from the European bee Apis mellifera (AmCA) spermatheca and inhibitory effects of pesticides belonging to various classes, such as carbamates, thiophosphates, and pyrethroids, were investigated herein. The inhibitory effects of methomyl, oxamyl, deltamethrin, cypermethrin, dichlorodiphenyltrichloroethane (DDT) and diazinon on AmCA were analysed. These pesticides showed effective in vitro inhibition of the enzyme, at sub-micromolar levels. The IC50 values for these pesticides ranged between of 0.0023 and 0.0385 µM. The CA inhibition mechanism with these compounds is unknown at the moment, but most of them contain ester functionalities which may be hydrolysed by the enzyme with the formation of intermediates that can either react with amino acid residues or bid to the zinc ion from the active site.

Pyrethroid and carbamate resistance in Australian Helicoverpa armigera (Lepidoptera: Noctuidae) from 2008 to 2015: what has changed since the introduction of Bt cotton?

Pyrethroid and carbamate resistance was evaluated in Helicoverpa armigera from 2008 to 2015. Insects were collected as eggs primarily from cultivated hosts in the major cropping areas of New South Wales and Queensland, Australia. Larvae reared from eggs were tested for resistance to fenvalerate, bifenthrin or methomyl in the F0 generation using a topical application of a discriminating dose of insecticide. In 2008-2009, resistance to fenvalerate was 71% and no resistance to bifenthrin was recorded. In the following two seasons, resistance to pyrethroids was relatively stable with fenvalerate resistance ranging from 63% to 67% and bifenthrin resistance ranging from 5.6% and 6.4% in 2009-2010 and 2010-2011, respectively. However, in 2011-2012, pyrethroid resistance had increased to 91% and 36% for fenvalerate and bifenthrin, respectively. Resistance remained above 90% for fenvalerate and above 35% for bifenthrin in the following three seasons from 2012 to 2015. In 2008-2009, methomyl resistance was 33% and declined to 22% and 15% in 2009-2010 and 2010-2011, respectively. Methomyl resistance remained at moderate levels from 2011-12 to 2014-15, ranging from 21% to 40%. Factors that influenced selection pressure of pyrethroid and carbamate insecticides and impacted resistance frequency in H. armigera may have been associated with changes in the composition of the cropping landscape. The rapid expansion of the pulse industry and the commensurate increased use of insecticide may have played a role in reselection of high-level pyrethroid resistance, and highlights the need for an urgent and strategic response to insecticide resistance management in the Australian grains industry.

Study of Synergism, Antagonism, and Resistance Mechanisms in Insecticide-Resistant Oxycarenus hyalinipennis (Hemiptera: Lygaeidae).

Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae) is an economic and key pest of the Malvaceae family widely distributed in the world. Significant field resistance to cypermethrin (26.69-fold), chlorpyrifos (32.60-fold), methomyl (10.87), acetamiprid (20.63-fold), fipronil (5.84-fold), and spirotetramat (116.02-fold) has been reported. Cypermethrin combined with methomyl and spirotetramat, methomyl with spirotetramat, acetamiprid with spirotetramat, and fipronil with spirotetramat had synergistic effects (combination index, (CI) in a laboratory population of O. hyalinipennis named Lab-PK. Methomyl combined with acetamiprid and acetamiprid with fipronil had synergistic effects on O. hyalinipennis in a field in Multan named Field-POP. Cypermethrin combined with methomyl and spirotetramat; chlorpyrifos with methomyl, acetamiprid, and spirotetramat; methomyl with acetamiprid and spirotetramat; and fipronil with spirotetramat also had synergistic effects on O. hyalinipennis in Field-POP. Enzyme inhibitors piperonyl butoxide and S,S,S-tri-n-butyl phosphorotrithioate significantly increased the toxicity of chlorpyrifos, methomyl, acetamiprid, and spirotetramat to O. hyalinipennis in Field-POP, suggesting a monooxygenase- and esterase-based resistance mechanism. However, fipronil did not synergize with PBO and DEF. This study suggests that insecticide mixtures showing synergism must be determined for insecticide resistance management and other strategies such as rotations, mosaics, and cultural control should also be considered for the management of O. hyalinipennis.

Acetylcholinesterases from entomopathogenic nematode Heterorhabditid bacteriophora: Susceptibility to insecticides and immunological characteristics.

Acetylcholinesterases (AChEs) from the infective juveniles (IJs) of entomopathogenic nematode (EPN) have been investigated with respect to their susceptibility to insecticides and immunological characteristics, aiming at nominating the most compatible insecticide(s) to be used in conjunction with the most insecticide-tolerant EPN strain before incorporation in integrated pest management (IPM) programs. The inhibition kinetics of two purified AChE isoenzymes, AChEAII and AChEBI isolated from Heterorhabditid bacteriophora EM2 strain, by different insecticides revealed that the insensitivity to inhibition by such insecticides could be arranged in a descending order as; methomyl>carbofuran>acetamiprid>oxamyl>malathion. Except for malathion, the insecticides competitively inhibited AChEs with Ki values ranging from 0.1 to 15mM and IC50 values from 1.25 to 23mM. The two AChE isoforms are several folds less sensitive to inhibition by methomyl and carbofuran compared to those previously reported for other insect species. AChEBI was used as an immunogen to raise anti-AChEBI antisera in rabbits. The prepared antisera cross-reacted with AChEs of five different heterorhabditid nematode strains implying the presence of common epitopes shared along all the examined strains. Such studies could aid in the rational selection of the compatible insecticide(s) and the prepared polyclonal anti-AChE antisera would be a valuable immunodiagnostic tool for evaluating the most insecticide-tolerant EPN strain(s) in IPM programs.

Novel Insights in the Regulation of Phosphatidylserine Exposure in Human Red Blood Cells.

BACKGROUND/AIMS: In previous publications we were able to demonstrate the exposure of phosphatidylserine (PS) in the outer membrane leaflet after activation of red blood cells (RBCs) by lysophosphatidic acid (LPA), phorbol-12 myristate-13acetate (PMA), or 4-bromo-A23187 (A23187). It has been concluded that three different mechanisms are responsible for the PS exposure in human RBCs: (i) Ca2+-stimulated scramblase activation (and flippase inhibition) by A23187, LPA, and PMA; (ii) PKCα activation by LPA and PMA; and (iii) enhanced lipid flip flop caused by LPA. Further studies aimed to elucidate interconnections between the increased Ca2+ content, scramblase- and PKCα-activation. In addition, the role of the Ca2+-activated K+ channel (Gardos channel) activity in the process of PS exposure needs to be investigated. METHODS: The intracellular Ca2+ content and the PS exposure of RBCs have been investigated after treatment with LPA (2.5 µM), PMA (6 µM), or A23187 (2 µM). Fluo-4 and annexin V-FITC has been used to detect intracellular Ca2+ content and PS exposure, respectively. Both parameters (Ca2+ content, PS exposure) were studied using flow cytometry. Inhibitors of the scramblase, the PKCα, and the Gardos channel have been applied. RESULTS: The percentage of RBCs showing PS exposure after activation with LPA, PMA, or A23187 is significantly reduced after inhibition of the scramblase using the specific inhibitor R5421 as well as after the inhibition of the PKCα using chelerythrine chloride or calphostin C. The inhibitory effect is more pronounced when the scramblase and the PKCα are inhibited simultaneously. Additionally, the inhibition of the Gardos channel using charybdotoxin resulted in a significant reduction of the percentage of RBCs showing PS exposure under all conditions measured. Similar results were obtained when the Gardos channel activity was suppressed by increased extracellular K+ content. CONCLUSION: PS exposure is mediated by the Ca2+-dependent scramblase but also by PKCα activated by LPA and PMA in a Ca2+-dependent and a Ca2+-independent manner. Furthermore, we hypothesize that a hyperpolarisation of RBCs caused by the opening of the Gardos channel is essential for the scramblase activity as well as for a fraction of the LPA-induced Ca2+ entry.

Behavioral and metabolic effects of sublethal doses of two insecticides, chlorpyrifos and methomyl, in the Egyptian cotton leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae).

Insecticides have long been used as the main method in limiting agricultural pests, but their widespread use has resulted in environmental pollution, development of resistances, and biodiversity reduction. The effects of insecticides at low residual doses on both the targeted crop pest species and beneficial insects have become a major concern. In particular, these low doses can induce unexpected positive (hormetic) effects on pest insects, such as surges in population growth exceeding what would have been observed without pesticide application. Methomyl and chlorpyrifos are two insecticides commonly used to control the population levels of the cotton leafworm Spodoptera littoralis, a major pest moth. The aim of the present study was to examine the effects of sublethal doses of these two pesticides, known to present a residual activity and persistence in the environment, on the moth physiology. Using a metabolomic approach, we showed that sublethal doses of methomyl and chlorpyrifos have a systemic effect on the treated insects. We also demonstrated a behavioral disruption of S. littoralis larvae exposed to sublethal doses of methomyl, whereas no effects were observed for the same doses of chlorpyrifos. Interestingly, we highlighted that sublethal doses of both pesticides did not induce a change in acetylcholinesterase activity in head of exposed larvae.

Inhibition kinetics of acid and alkaline phosphatases by atrazine and methomyl pesticides.

The main objective of this work was to investigate the kinetic characteristics of acid and alkaline phosphatases isolated from different sources and to study the effects of the herbicide atrazine and insecticide methomyl on the activity and kinetic properties of the enzymes. Acid phosphatase (ACP) was isolated from the tomato plant (Solanum lycopersicum L. var. lycopersicum); alkaline phosphatase (ALP) was isolated from two sources, including mature earthworms (Aporrectodea caliginosa) and larvae of the Egyptian cotton leafworm (Spodoptera littoralis). The specific activities of the enzymes were 33.31, 5.56 and 0.72 mmol substrate hydrolyzed per minute per milligram protein for plant ACP, earthworms ALP and cotton leafworm ALP, respectively. The inhibition kinetics indicated that atrazine and methomyl caused competitive-non-competitive inhibition of the enzymes. The relationships between estimates of K(m) and V(max) calculated from the Michaelis-Menten equation have been explored. The extent of the inhibition was different, as estimated by the values of the inhibition constant Ki that were found to be 3.34 × 10(-3), 1.12 × 10(-2) and 1.07 × 10(-2) mM for plant ACP, earthworms ALP and cotton leafworm ALP, respectively, with methomyl. In the case of atrazine, K(i) were found to be 8.99 × 10(-3), 3.55 × 10(-2) and 1.36 × 10(-2) mM for plant ACP, earthworms ALP and cotton leafworm ALP, respectively.

Digestive enzyme as benchmark for insecticide resistance development in Culex pipiens larvae to chemical and bacteriologic insecticides.

This work monitored changes in some digestive enzymes (trypsin and aminopeptidase) associated with the building up of resistance in Cx. pipiens larvae to two chemical insecticides (methomyl and/or malathion) and one biological insecticide (Bacillus thuringiensis-H14 or B.t H 14). The LC50 value of methomyl for both field- and the 12th generation (F12) of the selected strain was 1.789 ppm and 8.925 ppm respectively. The LC50 value of malathion for both field and the F12 of the selected strain was 0.082 ppm and 0.156 ppm respectively, and those of B.t H14 of field strain and the F12 was 2.550ppm & 2.395ppm respectively. The specific activity of trypsin enzyme in control susceptible colony was 20.806 +/- 0.452micromol/min/mg protein; but at F4 and F8 for malathion and methomyl treated larvae were 10.810 +/- 0.860 & 15.616+/-0.408 micromol/min/mg protein, respectively. Trypsin activity of F12 in treated larvae with B.t.H14 was 2.097 +/- 0.587 microiol/min/mg protein. Aminopeptidase specific activity for susceptible control larvae was 173.05 +/- 1.3111 micromol/min/mg protein. This activity decreased to 145.15 +/- 4.12, 152.497 +/- 6.775 & 102.04 +/- 3.58a micromol/min/mg protein after larval (F 12) treatment with methomyl, malathion and B.t H 14 respectively.

[Resistance mechanisms and cross-resistance of phoxim-resistant Frankliniella occidentalis Pergande population].

To understand the resistance risks of Frankliniella occidentalis Pergande against phoxim, this paper studied the resistance mechanisms of phoxim-resistant F. occidentalis population against phoxim and the cross-resistance of the population against other insecticides. The phoxim-resistant population had medium level cross-resistance to chlorpyrifos, lambda-cyhalothrin, and methomyl, low level cross-resistance to chlorfenapyr, imidacloprid, emamectin-benzoate, and spinosad, but no cross-resistance to acetamiprid and abamectin. The synergists piperonyl butoxide (PBO), s, s, s-tributyl phosphorotrithioate (DEF), and triphenyl phosphate (TPP) had significant synergism (P < 0.05) on the toxicity of phoxim to the resistant (XK), field (BJ), and susceptible (S) populations, while diethyl maleate (DEM) had no significant synergism to XK and S populations but had significant synergism to BJ population. As compared with S population, the XK and BJ populations had significantly increased activities of mixed-functional oxidases P450 (2.79-fold and 1.48-fold), b, (2.88-fold and 1.88-fold), O-demethylase (2.60-fold and 1.68-fold), and carboxylesterase (2.02-fold and 1.61-fold, respectively), and XK population had a significantly increased acetylcholine esterase activity (3.10-fold). Both XK and BJ population had an increased activity of glutathione S-transferases (1.11-fold and 1.20-fold, respectively), but the increment was not significant. The increased detoxification enzymes activities in F. occidentalis could play an important role in the resistance of the plant against phoxim.