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.

Lethal and sublethal effects of insecticides used in the management of Plutella xylostella (Lepidoptera: Plutellidae) on the predator Cycloneda sanguinea L. (Coleoptera: Coccinellidae).

BACKGROUND: The application of synthetic insecticides is the main strategy used to reduce the damage caused by the diamondback moth Plutella xylostella in commercial Brassica crops. However, incorrect insecticide use can cause biological and ecological disturbances in agroecosystems. Cycloneda sanguinea is a generalist voracious predator and is distributed widely in cultivated and noncultivated ecosystems. This study investigated the efficiency of four insecticides for the control of P. xylostella and the lethal and sublethal effects of these insecticides on C. sanguinea. RESULTS: Spinosad (92% mortality) and chlorfenapyr (76% mortality) were highly toxic to P. xylostela. However, chlorantraniliprole (10% mortality) and methomyl (no mortality) were ineffective against this pest. Chlorantraniliprole was the only insecticide that was highly toxic to C. sanguinea by contact (90% mortality), however, it was nontoxic following the ingestion of chlorantraniliprole-contaminated aphids. Interestingly, ingestion of prey contaminated with methomyl and chlorfenapyr was highly toxic (100% mortality) to C. sanguinea. Spinosad was nontoxic to C. sanguinea via exposure to contaminated surfaces and following ingestion of contaminated prey. However, direct contact of the insects with both methomyl and spinosad significantly affected C. sanguinea flight activity (vertical flight and free-fall flight), whereas chlorfenapyr impacted vertical flight only. CONCLUSION: These findings showed that chlorantraniliprole was not only ineffective for the control of P. xylostela, but was also highly toxic to C. sanguinea. The results indicated that spinosad was efficient against P. xylostela and was of low toxicity to C. sanguinea; however, the deleterious effects of this insecticide on flight behavior could result in reduced predatory efficiency. © 2022 Society of Chemical Industry.

Assessing the neurotoxicity of the carbamate methomyl in Caenorhabditis elegans with a multi-level approach.

The neurotoxicity and developmental effects of a widely applied insecticide (methomyl) was investigated by a multi-level approach (behavior and biometry, biochemical alterations and neurodegeneration) in Caenorhabditis elegans upon a short-term exposure (1 h) and a post-exposure period (48 h). The 1-h exposure to sub-lethal concentrations of methomyl (lower than 0.320 g L-1; i.e. below the estimated LC10) triggered significant changes on motor behavior and development impairment. The type of movement was significantly altered in methomyl-exposed worms, as well as biometric parameters (worms frequently idle and moving more backwards than controls; small body area, length and wavelength). These effects were followed by an increase of acetylcholine levels. Interestingly, after the 48-h recovery period, movement of previously exposed worms was similar to controls, and a concentration-dependent reversion of biometric endpoints was recorded, pointing out the transient action of the carbamate in line with an apparent absence of cholinergic neurons damage. This study provided new insight on the neurotoxicity of methomyl by showing that effects on movement and development were transient, and apparently did not result in neurodegeneration in cholinergic neurons. Moreover, these findings reinforced the advantages of using C. elegans in a multi-level approach for pesticide effects assessment.

Cytotoxicity and genotoxicity of methomyl, carbaryl, metalaxyl, and pendimethalin in human umbilical vein endothelial cells.

Pesticides have adverse effects on the cellular functionality, which may trigger myriad of health consequences. However, pesticides-mediated toxicity in the endothelial cells (ECs) is still elusive. Hence, in this study, we have used human umbilical vein endothelial cells (HUVECs) as a model to quantify the cytotoxicity and genotoxicity of four pesticides (methomyl, carbaryl, metalaxyl, and pendimethalin). In the MTT assay, HUVECs exposed to methomyl, carbaryl, metalaxyl, and pendimethalin demonstrated significant proliferation inhibition only at higher concentrations (500 and 1000 µM). Likewise, neutral red uptake (NRU) assay also showed proliferation inhibition of HUVECs at 500 and 1000 µM by the four pesticides, confirming lysosomal fragility. HUVECs exposed to the four pesticides significantly increased the level of intracellular reactive oxygen species (ROS). Comet assay and flow cytometric data exhibited DNA damage and apoptotic cell death in HUVECs after 24 h of exposure with methomyl, metalaxyl, carbaryl, and pendimethalin. This is a first study on HUVECs signifying the cytotoxic-genotoxic and apoptotic potential of carbamate insecticides (methomyl and carbaryl), fungicide (metalaxyl), and herbicide (pendimethalin). Overall, these pesticides may affect ECs functions and angiogenesis; nonetheless, mechanistic studies are warranted from the perspective of vascular biology using in vivo test models.

Simple Colorimetric Method for Cholinesterase-inhibitor Screening in Gastric Content by Using Phytoesterase Enzyme from Kidney Bean.

BACKGROUND AND OBJECTIVE: Diagnosis of cholinesterase inhibitor insecticide ingestion is based on clinical suspicious and should be confirmed by cholinesterase essay. However, serum cholinesterase activity test requires specific instruments and procedure. This study aimed to develop simple colorimetric test to detect cholinesterase inhibitors in the gastric content, using phytoesterase and alpha naphthyl acetate as a chromogenic substrate. MATERIALS AND METHODS: Methomyl and chlorpyrifos were selected for the phytoesterase enzyme inhibition assay. The experiment was conducted using pooled insecticide-free gastric content sample from ten cadavers. The gastric content samples were prepared by simple filtration procedure or liquid-liquid extraction procedure with dichloromethane or ethyl acetate. The inhibitor concentrations measured by the developed phytoesterase enzyme inhibition assay were compared with those analyzed by the LC-MS/MS and the GC-FPD. RESULTS: Different sample preparation procedures, sensitivity and specificity and specificity of the test were investigated. Sample extracted with dichloromethane reduced the effect of matrix in gastric content as same as ethyl acetate. The developed color test method of detection showed 56.52% sensitivity and 100% specificity for methomyl, 100% sensitivity and 96.30% specificity for chlorpyrifos. The limit of detection of the assay was 422.6 ng mL-1 for methomyl and was 339.8 ng mL-1 for chlorpyrifos. CONCLUSION: This developed method could be used an alternative diagnostic test for methomyl and chlorpyrifos self-ingestion.

Can Toxicities Induced by Insecticide Methomyl be Remediated Via Soil Bacteria Ochrobactrum thiophenivorans and Sphingomonas melonis?

The research study was about revealing the biochemical response of Gammarus pulex related to insecticide methomyl before and after bioremediation by two soil bacteria species, Ochrobactrum thiophenivorans and Sphingomonas melonis. Catalase (CAT), glutathione S-transferase.(GST), cytochrome. P4501A1 (CYP1A1) activities in G. Pulex related to methomyl solution were investigated in 24 h and 96 h. ELISA method was used for test studies. CAT enzyme was decreased in Gammarus pulex that was exposed to methomyl after all exposure period (P < 0.05). CAT activities were returned to control results after bioremediation assays. GST enzyme activity was decreased depending on methomyl exposure during 24 h but increased during 4 days (P < 0.05). After 8 days of bioremediation period, GST activity increased again during 24 h while decreased during 4 days (P < 0.05). CYP1A1 activity increased in Gammarus pulex that was exposed to methomyl after all exposure period (P > 0.05). After bioremediation, statistically significant changes were not revealed in CYP1A1 activities (P > 0.05). According to the results of our study, CYP1A1, CAT, and GST activities in G. pulex sanctioned the capability of Ochrobactrum thiophenivorans and Sphingomonas melonis in methomyl bioremediation. Isolated and enriched Ochrobactrum thiophenivorans and Sphingomonas melonis that were added to 2.5 ppb concentrations of methomyl for 8 days. Each day, chemical oxygen demand (COD) and biochemical oxygen demand (BOD5), pH and dissolved oxygen parameters were monitored. At the final phase of the bioremediation step, it was determined that these bacteria have efficient methomyl bioremediation properties in a mixed corsortia at a rate of 86%. These results show that these bacteria can be used for bioremediate the receiving environments that are polluted by these kinds of insecticides.

Genotoxic and mutagenic evaluation of the protective effect of exogenous melatonin in adult rats and their offspring exposed to the insecticides methomyl and cypermethrin during pregnancy.

Exposure of pregnant women to pesticides in an occupational or incidental manner has been pointed out as a possible cause of health problems in newborns. Cypermethrin and methomyl are insecticides whose active compounds and metabolites can cause harm to the health of pregnant women and their offspring. Both contribute to increases in the concentrations of oxidizing agents that damage DNA, causing mutations. Thus, the use of antioxidant substances, such as melatonin, could serve as protection against genetic damage, reducing the problems caused by insecticides. Here, we evaluated the protective effect of exogenous melatonin in adult rats and their offspring exposed to methomyl and cypermethrin during the gestational period. Thirty adult rats were divided into five groups from the first day of pregnancy: NC - negative control, Mth received 0.02 mg/kg/day of methomyl, Cyp received 0.05 mg/kg/day cypermethrin, MthMl received 0.02 mg/kg/day methomyl combined with 10 mg/kg/day melatonin, and CypMl received 0.05 mg/kg/day cypermethrin combined with 10 mg/kg/day melatonin. Blood and liver samples were collected from the dams; blood, liver, and brain samples were collected from the offspring. All tissues were used for comet assay; blood was used for micronucleus test. Our results demonstrate that cypermethrin and methomyl induce significant genetic damage in the tissues of dams and offspring. Treatment with melatonin significantly reduced cypermethrin-induced genetic damage in dams and pups; however, melatonin was not effective against the genetic damage caused by methomyl. Thus, melatonin may be considered a promising agent for the reduction of genotoxic effects induced by cypermethrin but appears to be inefficient against the genotoxic effects caused by methomyl, probably due to differences in the metabolism of each pesticide.

Induction of hyperpigmentation and heat shock protein 70 response to the toxicity of methomyl insecticide during the organ development of the Arabian toad, Bufo arabicus (Heyden,1827).

Methomyl (MET) is a carbamate insecticide which is used as a substitute for organophosphorus compounds to protect crops against insects. The present study aims to evaluate the cytoprotection response of pigment cells and heat shock protein 70 (HSP70) after exposure to MET during the tadpole developmental stages of the Arabian toad, Bufo arabicus. Three developmental larval stages of the toad were selected and divided into two groups; Control and MET-exposed (MET-EX) tadpoles (10ppm). MET-EX tadpoles showed an increased number of pigment cells in the liver, kidney, anterior eye chamber, and skin tissues as compared to the control. The glycogen content in the developing liver and muscles (myotomes) of MET-EX tadpoles was decreased as compared to the control. In the MET-EX tadpoles, immunohistochemical staining showed an increase of HSP70 expression in the liver hepatocytes, the nucleated red blood cells (nRBC) in kidney glomeruli, the iridocorneal angle of anterior eye chamber, and the skin as compared to the control. The current study concluded that pigment cells and HSP70 represented a cytoprotecting response against MET insecticide during the organ development of B. arabicas tadpoles. Therefore, MET use should be regularly monitored in the environment to protect animals and human from exposure to this insecticide.

Soluble esterases as biomarkers of neurotoxic compounds in the widespread serpulid Ficopomatus enigmaticus (Fauvel, 1923).

The characterization of soluble cholinesterases (ChEs) together with carboxylesterases (CEs) in Ficopomatus enigmaticus as suitable biomarkers of neurotoxicity was the main aim of this study. ChEs of F. enigmaticus were characterized considering enzymatic activity, substrate affinity (acetyl-, butyryl-, propionylthiocholine), kinetic parameters (Km and Vmax) and in vitro response to model inhibitors (eserine hemisulfate, iso-OMPA, BW284C51), and carbamates (carbofuran, methomyl, aldicarb, and carbaryl). CEs were characterized based on enzymatic activity, kinetic parameters and in vitro response to carbamates (carbofuran, methomyl, aldicarb, and carbaryl). Results showed that cholinesterases from F. enigmaticus showed a substrate preference for acetylthiocholine followed by propionylthiocholine; butyrylthioline was not hydrolyzed differently from other Annelida species. CE activity was in the same range of cholinesterase activity with acetylthiocholine as substrate; the enzyme activity showed high affinity for the substrate p-nytrophenyl butyrate. Carbamates inhibited ChE activity with propionylthiocholine as substrate to a higher extent than with acetylthiocoline. Also CE activity was inhibited by all tested carbamates except carbaryl. In vitro data highlighted the presence of active forms of ChEs and CEs in F. enigmaticus that could potentially be inhibited by pesticides at environmentally relevant concentration.

Plant allelochemicals affect tolerance of polyphagous lepidopteran pest Helicoverpa armigera (Hübner) against insecticides.

Cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), is a polyphagous lepidopteran pest distributed worldwide with a broad spectrum of host plants. However, the mechanism of H. armigera adaptation to various insecticides and defensive allelochemicals in its host plants is not fully understood. Therefore, this study examined the influence of consumption of plant allelochemicals on larval tolerance to methomyl and chlorpyrifos insecticides in H. armigera and its possible mechanism. Twelve plant allelochemicals were screened to evaluate their effects on larval sensitivity to methomyl. Of which flavone, coumarin, DIMBOA (2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one) and visnagin significantly reduced larval sensitivity to methomyl. Application of cytochrome P450 inhibitor piperonyl butoxide (PBO) significantly increased the mortality of methomyl-treated larvae. In contrast, PBO addition significantly decreased the mortality of chlorpyrifos-treated larvae. Moreover, allelochemical consumption enhanced the activities of glutathione S-transferase, carboxylesterase, cytochrome P450 and acetylcholinesterase in the midgut and fat body. The qRT-PCR analysis confirms that P450 genes, CYP6B2, CYP6B6 and CYP6B7 were induced by the four allelochemicals in the midguts and the fat bodies. In conclusion, the generalist H. armigera can take benefit of plant allelochemicals from its host plants to elaborate its defense against insecticides.

Teratogenicity and neurotoxicity effects induced by methomyl insecticide on the developmental stages of Bufo arabicus.

Methomyl (MET) is a carbamate insecticide that has been widely used to protect the crop against insects as an alternative for organophosphorus insecticide. Thus the present study aims to evaluate the potential toxic effects of MET on the developmental stages of Bufo arabicus. Tadpoles were classified into three stages (25, 37, 40). Every stage was divided into two groups, control and MET-treated group (10 ppm for two weeks) after LC50 determination in acute toxicity test for 96 h. Control and MET-treated larvae were examined at the level of morphological, histological, skeleton deformities and immunohistochemical labeling of alpha-synuclein in the spinal cord and dorsal root ganglion. MET-exposed larvae showed hyperactivity, extreme agitation, abnormal swimming and kinking tail as compared to control. Alizarin Red S-Alcian blue staining showed scoliosis in MET-treated tadpoles at 25 and 37 stages; kyphosis, retarded tail regression and reduced ossification of the phalanges of digits for both fore-and hind limbs were noted in MET-exposed tadpoles at 40 stage as compared to control. Histopathological changes in myotomes, notochord and spinal cord were shown in MET-exposed tadpoles as compared to control. Immunohistochemical examination showed an over expression of alpha-synuclein either in the neurons of the spinal cord or in the dorsal root ganglion of MET-exposed tadpoles at stage 40 as compared to control. The present study concluded that MET insecticide induces malformation and teratogenicity effects which were accompanied by neurodegenerative effects for the neurons either in the spinal cord or in the dorsal root ganglion.

Effect of Chronic Exposure to Methomyl on Tissue Damage and Apoptosis in Testis of Tilapia (Oreochromis niloticus) and Recovery Pattern.

Tilapia were exposed to 0, 0.2, 2, 20, 200 µg/L methomyl for 30 days, and then transferred to methomyl-free water for 18 days. Caspase-8 in serum, apoptosis rate, microstructure and ultra-microstructure of testis were checked after methomyl exposure and at 18 days after transferring to methomyl-free water. There were no significant changes in Caspase-8 activity, apoptosis rate, and tissue structure in testis exposed to 0.2 and 2 µg/L compared with control. However, when tilapia exposed to 20 and 200 µg/L, the Caspase-8 activity and apoptosis rate were induced significantly, and tissue damage happened compared with the control. Thus it would appear 2 µg/L methomyl might be considered as the no observed adverse effect level. Recovery data showed that the effects produced by lower concentration of 20 µg/L were reversible but not at the higher 200 µg/L concentration.

Methomyl induced effect on fortilin and S100A1 in serum and cardiac tissue: Potential biomarkers of toxicity.

Methomyl toxicity has been reported as a cause of several accidental and suicidal fatalities. The study is evaluating the effect of lethal methomyl toxicity on fortilin and S100A1 in serum and cardiac tissues. Adult 96 female Sprague-Dawley rats were divided equally into a control (euthanized by cervical dislocation) and a study group (overdosed with methomyl). The levels of fortilin and S100A1 in serum were measured antemortem (to establish the basal levels in serum) and postmortem (to evaluate changes after methomyl exposure) using enzyme-linked immunoassay. S100A1 was immunostained in sections from cardiac tissues. Both proteins in the control were not significantly different (p > 0.05) compared with the antemortem levels. On the contrast, both biomarkers levels in the intoxicated group were remarkably higher (p < 0.001) than the control and the antemortem levels. Ventricular tissues from the intoxicated rats presented depleted S100A1 immunostain in cardiomyocytes localized mainly in the epicardium with deeply stained adjacent cardiac fibroblasts. The cardiomyocytes were damaged with a prominent loss of striations compared to normal cardiac tissues from the control. The present outcomes explain to a certain degree the potential toxic effect of methomyl poisoning on the cardiac tissue. Both proteins could be added to the currently available battery of markers for assessing methomyl toxicity.

Assessing interactive mixture toxicity of carbamate and organophosphorus insecticides in the yabby (Cherax destructor).

Carbamate (CB) and organophosphorus (OP) pesticides are commonly detected in aquatic ecosystems and predominantly occur as mixtures of varying complexity. These pesticides inhibit the activity of total cholinesterase (ChE) and thus have the potential to interfere with behaviours that may be essential for the survival of aquatic species. Although the effects of individual ChE insecticides on aquatic species have been reported for decades, the neurotoxicity of mixtures is still poorly understood. This study examined the chronic toxicities of two OP insecticides (chlorpyrifos (CPF) and malathion (MAL)) and one carbamate insecticide (methomyl (METH)) in binary and ternary mixtures on the ChE activity of the yabby (C. destructor). Using the concentration addition approach to estimate mixture toxicity, the observed inhibition of ChE activity caused by all binary mixtures of CPF plus MAL, CPF plus METH and MAL plus METH was additive. In ternary mixtures, all combinations of CPF, MAL and METH were either additive or antagonistic depending on the relative ratios of these chemicals in the mixtures. The effect of mixtures of these three insecticides on C. destructor has not previously been assessed, and the data suggest that individual chemical risk assessments are likely to incorrectly estimate the effect of these insecticides on C. destructor in the aquatic environment where combinations of such chemicals occur.

Chlorantraniliprole as a candidate pesticide used in combination with the attracticides for lepidopteran moths.

Methomyl is currently used as a toxicant for the attracticide BioAttract in cotton and vegetables in China. However, methomyl is highly toxic to non-target organisms and a more environmental friendly acceptable alternative is required. Larvae of three lepidopteran insects Helicoverpa armigera, Agrotis ipsilon and Spodoptera litura are important pests of these crops in China. In the present study, the toxicity of 23 commonly used insecticides were tested on H. armigera, then tested the susceptibility of A. ipsilon and S. litura moths to the insecticides which were the most toxic to H. armigera, and the acute toxicity of the most efficacious insecticides were further investigated under laboratory conditions. Chlorantraniliprole, emamectin benzoate, spinetoram, spinosad and methomyl exhibited high levels of toxicity to H. armigera moths with a mortality of 86.67%, 91.11%, 73.33%, 57.78% and 80.00%, respectively, during 24 h period at the concentration of 1 mg a.i. L-1. Among these five insecticides, A. ipsilon and S. litura moths were more sensitive to chlorantraniliprole, emamectin benzoate and methomyl. The lethal time (LT50) values of chlorantraniliprole and methomyl were shorter than emamectin benzoate for all three lepidopteran moth species at 1000 mg a.i. L-1 compared to concentrations of 500, 100 and 1 mg a.i L-1. Chlorantraniliprole was found to have similar levels of toxicity and lethal time on the three lepidopteran moths tested to the standard methomyl, and therefore, can be used as an alternative insecticide to methomyl in the attracticide for controlling these pest species.

Evaluating the non-lethal effects of organophosphorous and carbamate insecticides on the yabby (Cherax destructor) using cholinesterase (AChE, BChE), Glutathione S-Transferase and ATPase as biomarkers.

The toxicity of two organophosphorus insecticides, chlorpyrifos (CPF), malathion (MAL), and one carbamate insecticide, methomyl (METH), to the yabby (Cherax destructor) was assessed by measuring cholinesterase (AChE, BChE), Glutathione S-Transferase (GST) and Na+/K+ATPase activity after 96h of exposure. Yabbies exposed to all three insecticides at 2 and 5µgL-1 exhibited significant AChE, BChE, GST and Na+/K+ATPase inhibition. Based on these enzyme inhibition tests, the toxicity of the three insecticides to C. destructor was CPF > MAL > METH. After 14 days of recovery the yabbies enzymatic activities of AChE, BChE, GST and Na+/K+ATPase was measured. Recovery of The enzyme activity recovery was faster after the exposure to METH than for the yabbies exposed to CPF and MAL. Slow recovery of enzyme activity could affect the physical activities of organisms and produce indirect effects on populations if such crayfish are less able to elude predators or search for food.

Effect of methomyl on sex steroid hormone and vitellogenin levels in serum of male tilapia (Oreochromis niloticus) and recovery pattern.

Tilapia were exposed to sub-lethal concentrations of 0, 0.2, 2, 20 or 200 µg/L for 30 days, then transferred to methomyl-free water for 18 days. E2 , T, 11-KTand VTG in serum were examined. There were no significant changes in all the parameters in serum of tilapia exposed to 0.2 µg/L and 2 µg/L methomyl compared to the control. However, 20 µg/L and 200 µg/L have the potential to disrupt the endocrine system of male tilapia, as shown by its ability to increase VTG and E2 and decrease T and 11-KT in serum. Thus it would appear the no observed adverse effect level for sexual steroid hormones of methomyl is lower than 2 µg/L. Recovery data showed that the effects produced by 20µg/L were reversible but not at 200µg/L. Furthermore, the sensitivity of above parameters to methomyl followed the order of VTG>E2 >11-KT>T>GSI, suggesting VTG being the better biomarkers.

A comparative assessment of cytotoxicity of commonly used agricultural insecticides to human and insect cells.

The cytotoxic potential of 13 commonly used agricultural insecticides was examined using cell-based systems with three human HepG2, Hek293, HeLa cells and three insect Tn5B1-4, Sf-21, and Drosophila S2 cells. Data showed that (1) an enhancement of some insecticides (e.g. pyrethroids) on cells proliferation; (2) an inhibition of some insecticides on cells viability; (3) various levels of susceptibility of different cells to the same insecticide; and (4) the cell type dependent sensitivity to different insecticides. The degree of cytotoxicity of insecticides on human cells was significantly lower than that on insect cells (P<0.05). Methomyl, even 20µg/ml, showed little cytotoxicity at 24h exposure whereas emamectin benzoate possessed the strongest cytotoxic potential in a dose-dependent fashion. The results revealed comparable cytotoxic property of agricultural insecticides against intact cells.

Behavior and Survival of the Filth Fly Parasitoids Spalangia endius and Urolepis rufipes (Hymenoptera: Pteromalidae) in Response to Three Granular House Fly Baits and Components.

Behaviors and mortality of two filth fly parasitoid wasps, Spalangia endius Walker and Urolepis rufipes Ashmead, were tested in response to granular fly baits containing one of the three active ingredients (AI): Golden Malrin (methomyl), QuickBayt (imidacloprid), or Quikstrike (dinotefuran). Behavioral responses to each of the two components of the baits, the AIs and the fly attractant pheromone (Z)-9-tricosene, were also examined independently. Spalangia endius avoided contact with bait granules, regardless of bait type. However, when S. endius contacted bait residue, the imidacloprid bait appeared to be the least harmful of the baits for S. endius, at least in the short term. Spalangia endius was attracted to imidacloprid by itself. However, S. endius avoided (Z)-9-tricosene. In contrast to S. endius' attraction to imidacloprid, S. endius neither avoided nor was attracted to methomyl or dinotefuran. For U. rufipes, the methomyl bait appeared to be especially harmful. Urolepis rufipes avoided bait granules with imidacloprid or dinotefuran but not with methomyl, died quickly in the presence of methomyl bait residue, and had a methomyl LC50 that was lower than that for S. endius The avoidance by U. rufipes of granules with imidacloprid or dinotefuran appears to be related to components other than the AIs or the (Z)-9-tricosene because U. rufipes did not avoid either individually. The behavioral avoidance of the parasitoids in the present study occurred despite no exposure recently, if ever, to these pesticides.

1H-NMR-based metabolomic study on toxicity of methomyl and methidathion in fish.

A 1H-nuclear magnetic resonance (NMR) spectroscopy with multivariate analysis was applied to detect the toxicity of antiacetylcholinesterase insecticides, methomyl (methyl (1E)-N-(methylcarbamoyloxy)ethanimidothioate) and methidathion (3-(dimethoxyphosphinothioyl sulfanylmethyl)-5-methoxy-1,3,4-thiadiazol-2-one), using zebrafish (Danio rerio) and Chinese bleak (Aphyocypris chinensis). Generally, methomyl and methidathion have been believed not to highly accumulate in fish tissues. However, these pesticides showed their toxicity by altering patterns of whole-body metabolites in neurotransmitter balance, energy metabolism, oxidative stress, and muscle maintenance in low concentrations. We used Pearson correlation analysis to contextualize the metabolic markers in pesticide treated groups. We observed that the positive correlations of choline with acetate and betaine in untreated control were shifted to null correlations showing acetylcholinesterase specific toxicity. This research demonstrated the applicability and potential of NMR metabolomics in detecting toxic effects of insecticide with a modicum of concentrations in aquatic environment.