Anticholinesterase Insecticides Research Articles

Human blood markers of cholinergic neurotoxicity and neuropathy: A useful guide for laboratory applications

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are useful markers to assess the effects of exposure to anticholinesterase insecticides (Anti-AChE). In addition, lymphocyte neuropathy target esterase (LNTE) has been used as biomarker of neuropathic organophosphate compounds (OPs). Thus, this study evaluates the main types of circulating biomarkers related to the cholinergic system and to the neuropathy induced by OPs in standardized human samples. To achieve this objective, total protein of human plasma, erythrocytes and lymphocytes were first standardized, and then AChE, BChE and LNTE activities in human blood were evaluated in the presence of inhibitors. The acceptance criteria of the regulatory agency were respected with coefficients of regression of curves of 0.9972 for cholinesterase and 0.9956 for LNTE analyses. The wavelength established to perform cholinesterase assay was 450 nm and the time of incubation of the enzymes with inhibitors was 30 min. Differences were observed among the IC50 values regarding the in vitro inhibition of AChE, BChE and LNTE in the presence of OPs. In conclusion, the procedures demonstrated by the present work were simple, fast, inexpensive, sensitive, easy to be replicated and suitable to make conclusions about the neurotoxicity induced by Anti-AChE and neuropathic OPs.

Effectiveness of calcium channel blockade for organophosphorus and carbamate pesticide poisoning – study protocol for an open label, pragmatic, 3-arm RCT repurposing two widely available medicines

Pesticide self-poisoning is one of the three most important global means of suicide, killing an estimated 110–168,000 people each year, mostly in poor rural Asian communities. Organophosphorus (OP) and carbamate anticholinesterase insecticides are responsible for about two-thirds of these deaths. Calcium channel blocking medicines (CCB) may reduce the effect of pesticides and prevent deaths. Two preclinical rodents’ studies and eight clinical studies utilising nimodipine and magnesium sulphate (MgSO4), respectively, showed mixed results. We have established a multi-centre randomised controlled trial (RCT) of patients with OP or carbamate self-poisoning admitted to at least six major hospitals in Bangladesh. The study aims to recruit maximum 3,243 patients over four years. One-third of the patients selected at random will receive standard treatment, while one-third will be treated with additional nimodipine and one-third with additional MgSO4. The additional treatments will be given for 48 h. We will check mortality (currently an estimated 11% die with standard treatment) and need for intensive care for mechanical ventilation across the three groups. This could lead to development of the first novel treatment for anticholinesterase poisoning in 50 years and its introduction into routine hospital practice worldwide.

Discovery selective acetylcholinesterase inhibitors to control Tetranychus urticae (Acari: Tetranychidae).

The two-spotted spider mite, Tetranychus urticae Koch, has a broad host plant range and presents an extreme capacity for developing pesticide resistance, becoming a major economic pest in agriculture. Anticholinesterase insecticides still account for a big part of global insecticide sales. However, there is a growing concern about the serious resistance problems of anticholinesterase insecticides and their nontarget toxicity. In this study, structure-based virtual screening was performed to discover selective AChE inhibitors from the ChemBridge database, and 39 potential species-specific AChE inhibitor were obtained targeting T. urticae AChE, but not human AChE. Among them, compound No. 8 inhibited AChE from T. urticae, but not from human, and had an inhibitory activity comparable to that of eserine. Compound No. 8 had dose-dependent toxicity to T. urticae in glass slide-dipping assay and had significant mite control effects in a pot experiment, but required a high concentration to achieve similar control effects to spirodiclofen. The toxicity evaluation suggested that compound No. 8 had no acute toxicity on pollinator honey bees and natural predator N. californicus and did not affect strawberry growth in our assay. Compound No. 8 is a potential lead compound for developing novel acaricides with reduced nontarget toxicity.

Larvicidal activity assessment for anticholinesterase insecticides against laboratory and field strains of Egyptian cotton leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)

Larvicidal activity assessment for anticholinesterase insecticides against laboratory and field strains of Egyptian cotton leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)

In vitro and in silico analysis of the Anopheles anticholinesterase activity of terpenoids.

Anopheles gambiae, An. coluzzii, An. arabiensis, and An. funestus are major vectors in high malaria endemic African regions. Various terpenoid classes form the main chemical constituent repository of essential oils, many of which have been shown to possess insecticidal effects against Anopheles species. The current study aimed to assess the bioactivity of terpenoids including four sesquiterpene alcohols, farnesol, (-)-α-bisabolol, cis-nerolidol, and trans-nerolidol; a phenylpropanoid, methyleugenol, and a monoterpene, (R)-(+)-limonene, using the larvicidal screening assay against the four Anopheles species. The mechanism of action was investigated through in vitro acetylcholinesterase inhibition assay and in silico molecular modelling. All six terpenoids showed potent larvicidal activity against the four Anopheles species. Insights into the mechanism of action revealed that the six terpenoids are strong AChE inhibitors against An. funestus and An. arabiensis, while there was a moderate inhibitory activity against An. gambiae AChE, but very weak activity against An. coluzzii. Interestingly, in the in silico study, farnesol established a favourable hydrogen bonding interaction with a conserved amino acid residue, Cys447, at the entrance to the active site gorge. While (-)-α-bisabolol and methyleugenol displayed a strong interaction with the catalytic Ser360 and adjacent amino acid residues; but sparing the mutable Gly280 residue that confers resistance to the current anticholinesterase insecticides. As a result, this study identified farnesol, (-)-α-bisabolol, and methyleugenol as selective bioinsecticidal agents with potent Anopheles AChE inhibition. These terpenoids present as natural compounds for further development as anticholinesterase bioinsecticides.

Blood cholinesterase activities and oxidative stress status among farmworkers using pesticides in Duhok, KRG, Iraq

Background: The use of pesticides by farmworkers poses considerable health risks. This study was undertaken to examine plasma and erythrocyte cholinesterase activities, plasma oxidative biomarkers malondialdehyde (MDA), and total antioxidant status (TAS) among farmworkers using different pesticide products in Duhok, northern of Iraq. Methods: This is a case-control study conducted between November 2021 to July 2022 on 92 male farmworkers who were exposed to pesticides in comparison with 44 non-exposed male subjects (control). The availability and uses of pesticides were obtained from 19 agrochemical shops and the farmworkers exposed to pesticides. Demographic data of pesticide-exposed farmworkers and their practice of pesticide applications were recorded. Plasma and erythrocyte cholinesterase activities and plasma MDA and TAS levels were determined in both groups. Results: The farmworkers had a significant 10.0% increase in plasma MDA level, with no significant changes in blood cholinesterase activities or the TAS level. Odds and risk ratios of reduced plasma cholinesterase activity (20.0%) suggested an association of health risks in pesticide-exposed farmworkers. Most of the pesticide products (278) in use were insecticides (47.0%), which comprised mainly 26.0% pyrethroids and 3.0-7.0% anticholinesterase insecticides, among others. The majority of the farmworkers (51%) were merely aware of the general target use of the pesticide, and 75% had an exposure history of > 5 years. Pesticide application was mostly (50.0%) manual, and 54.0% used insufficient personal protection equipment; 32.0% ate and drank at work, 48.0% practiced disposal of empty pesticide containers by burning and/or burying them, whereas 25.0% dumped the containers indiscriminately, and 25% disposed them at garbage sites openly. Conclusion: The farmworkers, with only a marginal increase in oxidative stress biomarker MDA, did not suffer from significant reductions in blood cholinesterase activities, although odds and risk ratios of reduced plasma cholinesterase activity suggested a health risk. Implementation of a national program is needed to measure pre-exposure blood cholinesterase activities in farmworkers.

The in silico and in vitro analysis of donepezil derivatives for Anopheles acetylcholinesterase inhibition.

Current studies on Anopheles anticholinesterase insecticides are focusing on identifying agents with high selectivity towards Anopheles over mammalian targets. Acetylcholinesterase (AChE) from electric eel is often used as the bioequivalent enzyme to study ligands designed for activity and inhibition in human. In this study, previously identified derivatives of a potent AChE, donepezil, that have exhibited low activity on electric eel AChE were assessed for potential AChE-based larvicidal effects on four African malaria vectors; An. funestus, An. arabiensis, An. gambiae and An. coluzzii. This led to the identification of four larvicidal agents with a lead molecule, 1-benzyl-N-(thiazol-2-yl) piperidine-4-carboxamide 2 showing selectivity for An. arabiensis as a larvicidal AChE agent. Differential activities of this molecule on An. arabiensis and electric eel AChE targets were studied through molecular modelling. Homology modelling was used to generate a three-dimensional structure of the An. arabiensis AChE for this binding assay. The conformation of this molecule and corresponding interactions with the AChE catalytic site was markedly different between the two targets. Assessment of the differences between the AChE binding sites from electric eel, human and Anopheles revealed that the electric eel and human AChE proteins were very similar. In contrast, Anopheles AChE had a smaller cysteine residue in place of bulky phenylalanine group at the entrance to the catalytic site, and a smaller aspartic acid residue at the base of the active site gorge, in place of the bulky tyrosine residues. Results from this study suggest that this difference affects the ligand orientation and corresponding interactions at the catalytic site. The lead molecule 2 also formed more favourable interactions with An. arabiensis AChE model than other Anopheles AChE targets, possibly explaining the observed selectivity among other assessed Anopheles species. This study suggests that 1-benzyl-N-(thiazol-2-yl) piperidine-4-carboxamide 2 may be a lead compound for designing novel insecticides against Anopheles vectors with reduced toxic potential on humans.

Potential of Essential Oil-Based Anticholinesterase Insecticides against Anopheles Vectors: A Review.

The insect nervous system is critical for its functional integrity. The cholinergic system, of which acetylcholinesterase (AChE) is a key enzyme, is essential to the Anopheles (consisting of major malaria vector species) nervous system. Furthermore, the nervous system is also the primary target site for insecticides used in malaria vector control programs. Insecticides, incorporated in insecticide-treated nets and used for indoor residual spraying, are a core intervention employed in malaria vector control. However, Anopheles resistance against these insecticides has grown rapidly. Due to this major setback, novel agents with potential activity against resistant Anopheles and/or capacity to overcome resistance against current WHO-approved insecticides are urgently needed. The essential oils have the potential to be natural sources of novel insecticides with potential to inhibit the Anopheles AChE target. In the current review, the scientific evidence highlights the ability of essential oils and specific essential oil constituents to serve as anticholinesterase insecticides. For this reason, the published data from scientific databases on the essential oils and essential oil constituents on anticholinesterase, ovicidal, larvicidal, pupicidal and adulticidal activities were analyzed. The identification of major constituents in active essential oils and their possible influence on the biological activity have also been critically evaluated. Furthermore, the toxicity to mammals as well as potential activity against the mammalian AChE target has also been reviewed. The importance of identifying novel potent insecticides from essential oils has been discussed, in relation to human safety and cost-effectiveness. Finally, the critical insights from this review can be used to inform future researchers towards potent and safe anticholinesterase insecticides for the management of Anopheles malaria vectors.

Non-covalent acetylcholinesterase inhibitors: In vitro screening and molecular modeling for novel selective insecticides

Insecticides represent the most crucial element in the integrated management approach to malaria and other vector-borne diseases. The evolution of insect resistance to long-used substances and the toxicity of organophosphates (OPs) and carbamates are the main factors contributing to the development of new, environmentally safe pesticides. In our work, fourteen compounds of 7-methoxytacrine-tacrine heterodimers were tested for their insecticidal effect. Compounds were evaluated in vitro on insect acetylcholinesterase from Anopheles gambiae (AgAChE) and Musca domestica (MdAChE). The evaluation was executed in parallel with testing on human erythrocyte acetylcholinesterase (HssAChE) and human butyrylcholinesterase (HssBChE) using a modified Ellman's method. Compound efficacy was determined as IC50 values for the respective enzymes and selectivity indexes were expressed to compare the interspecies selectivity. Docking studies were performed to predict the binding modes of selected compounds. K1328 and K1329 provided high HssAChE/AgAChE selectivity outperforming standard pesticides (carbofuran and bendiocarb), and thus can be considered as suitable lead structure for novel anticholinesterase insecticides.

Analytical and Bio-Analytical Method Development and Validation of Dichlorvos Pesticide Using RP-HPLC Method

Organophosphorus compounds were synthesised in the 1800s. Later they are used as insecticides in the late 1930s and early 1940s. The German scientist Gerhard Schrader is known for the creation of the basic chemical structure of anticholinesterase organophosphate compounds and development of the first commercialised Organophosperous insecticide. Such chemicals are anticholinesterase insecticide commonly used in agriculture and horticulture. To a lesser extent, they are used for domestic use. Due to the absence of bio persistence in organophosphates, most of the western countries opted to substitute organochlorines with organophosphates. Organophosphate pesticides are commonly used around the world, and contamination by these compounds is a serious public health concern in developing countries. Toxicokinetics and toxicodynamics of OP poisoning not only differ with path or level of exposure. But also the agent's chemical composition. Organophosphates are a group of pesticide that was developed in the 1940s in Germany and soon became an effective defence against agricultural pests. Dichlorvos which is a commonly used group of pesticide is a broad-spectrum organophosphate compound having insecticidal activity. Dichlorvos is a cholinesterase inhibitor exhibiting stomach, contact and systemic mode of action. Therefore, an accurate, fast, cost-effective and straightforward RP-HPLC technique for detecting Dichlorvos was developed. The RP-HPLC method is established by using ACN and Millipore water 50:50 v/v as mobile phase, the Flow rate is maintained at 1.5mL/minute. Detection of Dichlorvos was performed by using a PDA detector at 200nm. By this RP-HPLC procedure, RT of Dichlorvos was identified at 2.9 min.

Sublethal neurotoxicity of organophosphate insecticides to juvenile coho salmon

For decades, organophosphate (OP) insecticides have been used as chemical control agents in watersheds that support at-risk populations of Pacific salmon throughout western North America. Spray drift, runoff, and other processes transport OPs to critical surface water habitats for migratory salmonids. While most OPs share a common mechanism of action (i.e., inhibition of neuronal acetylcholinesterase, or AChE), they typically vary in toxic potency. Moreover, dose-response relationships for exposure and sublethal neurotoxicity (e.g., brain AChE inhibition) in salmonids have not been defined for many OPs. Here we exposed juvenile coho salmon (Oncorhynchus kisutch) to five common anticholinesterase insecticides (dimethoate, ethoprop, naled, phorate and phosmet) that are widely used on agricultural, commercial, residential, and public lands. Each of the five pesticides produced a concentration-dependent inhibition of AChE enzyme activity. The effective concentration for 50 % AChE inhibition (96-hr EC50) indicated the highest toxicity for phorate (EC50 = 0.57 μg/L) followed by phosmet (3.3 μg/L), naled (7.8 μg/L), ethoprop (90.6 μg/L) and dimethoate (273 μg/L). These findings can inform 1) relative hazard analyses for OP use near sensitive aquatic habitats, 2) predictions of sublethal OP mixture toxicity, and 3) ecological risk assessments for threatened or endangered species of Pacific salmon.

VARIACIÓN DE LOS NIVELES DE BUTIRILCOLINESTERASA DE UNA POBLACIÓN MEXICANA EN DISTINTOS PERIODOS DE CONTROL DE Aedes SPP.

The impact of organophosphate and carbamate pesticides used to control the Aedes spp. mosquito in sanitary campaigns was determined by measuring serum butirylcholinesterase activity in dry and rainy seasons. Differences in levels and their trend over time were analyzed, finding statistically significant differences between both seasons (men: dry season = 5272 U/L; rainy season = 4854 U/L; p < 0.05), with a decrease of 92 U/L each month. The decrease in men was 92 U/L each month. The enzymatic inhibition may be due to exposure to these anticholinesterase insecticides, and protective measures must be taken against the related health risks.

Latrophilin mediates insecticides susceptibility and fecundity through two carboxylesterases, esterase4 and esterase6, in Tribolium castaneum.

Latrophilin (LPH) is known as an adhesion G-protein-coupled receptor which involved in multiple physiological processes in organisms. Previous studies showed that lph not only involved the susceptibility to anticholinesterase insecticides but also affected fecundity in Tribolium castaneum. However, its regulatory mechanisms in these biological processes are still not clear. Here, we identified two potential downstream carboxylesterase (cce) genes of Tclph, esterase4 and esterase6, and further characterized their interactions with Tclph. After treatment of T. castaneum larvae with carbofuran or dichlorvos insecticides, the transcript levels of Tcest4 and Tcest6 were significantly induced from 12 to 72 h. RNAi against Tcest4 or Tcest6 led to the higher mortality compared with the controls after the insecticides treatment, suggesting that these two genes play a vital role in detoxification of insecticides in T. castaneum. Furthermore, with insecticides exposure to Tclph knockdown beetles, the expression of Tcest4 was upregulated but Tcest6 was downregulated, indicating that beetles existed a compensatory response against the insecticides. Additionally, RNAi of Tcest6 resulted in 43% reductions in female egg laying and completely inhibited egg hatching, which showed the similar phenotype as that of Tclph knockdown. These results indicated that Tclph affected fecundity by positively regulating Tcest6 expression. Our findings will provide a new insight into the molecular mechanisms of Tclph involved in physiological functions in T. castaneum.

Reference values for acetyl and butyrylcholinesterases in cattle under actual management conditions, hepatic and renal function by application of chlorpyrifos

ABSTRACTChlorpyrifos is an anticholinesterase organophosphate insecticide widely used in Argentina in the production of food derived from animal, fruit and horticultural origin and is reported as a residue within these products. Local reference values for acetyl and butyrylcholinesterase were determined in Aberdeen Angus bovine and cross bred cattle (n = 25), a requirement to be able to evaluate toxicity of commercial organophosphate and carbamate formulations. The activity of cholinesterase enzymes presented an overall mean of 2,183.00 ± 485.6 IU L−1 for erythrocyte acetylcholinesterase and 203.1 ± 42.06 IU L−1 for plasma butyrylcholinesterase, which are used as reference values for meat steers within a system of intensive production in a semi-arid region. The toxic potential of chlorpyrifos in steers of the same breeds (n = 12) was assessed applying chlorpyrifos 15.00% Tipertox® in a single therapeutic dose of 7.50 mg kg−1 by topical route. Prior to application and then on day 1 and day 21 post-application, both blood cholinesterases, serum chlorpyrifos concentration by ultra-high resolution liquid chromatography with mass detector, analysis of blood counts, total proteins, liver enzymes, urea and creatinine were evaluated. The mean plasma concentration of chlorpyrifos was 27.90 ug L−1 at 24 h. The findings indicate that the therapeutic treatment of castrated male bovines treated with chlorpyrifos, applied by pour-on according to the manufacturer's instructions, does not cause changes in the variables evaluated.

Structure of the G119S Mutant Acetylcholinesterase of the Malaria Vector Anopheles gambiae Reveals Basis of Insecticide Resistance

Malaria is a devastating disease in sub-Saharan Africa and is transmitted by the mosquito Anopheles gambiae. While indoor residual spraying of anticholinesterase insecticides has been useful in controlling the spread of malaria, widespread application of these compounds has led to the rise of an insecticide-resistant mosquito strain that harbors a G119S mutation in the nervous system target enzyme acetylcholinesterase. We demonstrate the atomic basis of insecticide resistance through structure determination of the G119S mutant acetylcholinesterase of An. gambiae in the ligand-free state and bound to a potent difluoromethyl ketone inhibitor. These structures reveal specific features within the active-site gorge distinct from human acetylcholinesterase, including an open channel at the base of the gorge, and provide a means for improving species selectivity in the rational design of improved insecticides for malaria vector control.

Reference intervals for B-esterases in gull, Larus michahellis (Nauman, 1840) from Northwest Spain: influence of age, gender, and tissue.

Over the last years, cholinesterase (ChE) and carboxylesterase (CbE) activities have been increasingly used in environmental biomonitoring to detect the exposure to anticholinesterase insecticides such as organophosphorates (OPs) and carbamates (CBs). The aim of this study was to determine ChE and CbE enzymatic activities present in liver and muscle of yellow-legged gulls (Larus michahellis), a seabird species considered suitable to monitor environmental pollution. In order to provide reference data for further biomonitoring studies, the influence of different factors, such as gender, age, sampling mode, and tissue, was considered in the present study. Our data report a statistically significant difference in CbE enzymatic activity comparing liver and muscle samples (P<0.05) along with an age-related CbE activity in liver samples (P<0.05). Moreover, according to our results, capture method might influence CbE and ChE activity in both liver and muscle samples (P<0.05). These findings underline the importance to assess basal levels of ChE and CbE activity considering, among other factors, gender-, age- and organ-related differences and confirm the suitability of Larus michahellis as a sentinel species especially within an urban environment.

Discovery of Species-selective and Resistance-breaking Anticholinesterase Insecticides for the Malaria Mosquito

Great reductions in malaria mortality have been accomplished in the last 15 years, in part due to the widespread roll-out of insecticide-treated bednets across sub-Saharan Africa. To date, these nets only employ pyrethroids, insecticides that target the voltage-gated sodium ion channel of the malaria vector, Anopheles gambiae. Due to the growing emergence of An. gambiae strains that are resistant to pyrethroids, there is an urgent need to develop new public health insecticides that engage a different target and possess low mammalian toxicity. In this review, we will describe efforts to develop highly species-specific and resistance-breaking inhibitors of An. gambiae acetylcholinesterase (AgAChE). These efforts have been greatly aided by advances in knowledge of the structure of the enzyme, and two major inhibitor design strategies have been explored. Since AgAChE possesses an unpaired Cys residue not present in mammalian AChE, a logical strategy to achieve selective inhibition involves design of compounds that could ligate that Cys. A second strategy involves the design of new molecules to target the catalytic serine of the enzyme. Here the challenge is not only to achieve high inhibition selectivity vs human AChE, but also to demonstrate toxicity to An. gambiae that carry the G119S resistance mutation of AgAChE. The advances made and challenges remaining will be presented. This review is part of the special issue "Insecticide Mode of Action: From Insect to Mammalian Toxicity".

Pancreatic Dysfunction Associated With Severe Acute Anticholinesterase Insecticide Poisoning In Adults

Background: The current study was carried out to study the effect of severe acute organophosphate compounds (OP) toxicity on the pancreatic profile in adults. Subjects and Methods: Six ml of venous blood were drawn from 260 patients (18-52 years) with severe acute OP toxicity who were admitted to the intensive care unit of Minia Poisoning Control Centre (PCC) during the period from May the 1st, 2012 to April, the 30th, 2014, and also from 20 healthy adults of the same age group acting as control. Blood samples were analyzed for: Pseudocholinestrase (PChE), serum insulin, serum α-amylase and serum lipase. Results: PChE level in intoxicated patients on the day of admission was highly significant decreased when compared with control. There was highly significant increase in the serum levels of amylase and lipase in intoxicated patients as compared to normal control group. In addition, there was highly significant increase in blood glucose level in intoxicated patients as compared with control group while there was no significant effect on the serum insulin level. A significant correlation was found between the degree of PChE inhibition and the elevation of serum amylase, lipase and serum glucose level. Conclusion: acute pancreatitis was evident in severe cases of OP intoxication on the day of admission, also OP -induced hyperglycemia is evident and most probably due to other cause rather than hypoinsulinemia.

Binary combinations of organophosphorus and synthetic pyrethroids are more potent acetylcholinesterase inhibitors than organophosphorus and carbamate mixtures: An in vitro assessment

Anticholinesterase insecticides such as organophosphorous (OP) and carbamates pesticides (CB); and synthetic pyrethroids (SP) pesticides commonly co-occur in the environment. This raises the possibility of antagonistic, additive, or synergistic neurotoxicity in exposed organisms. Acetylcholinesterase (AChE) inhibition has been demonstrated to be useful as a biomarker for exposure to OP and CBs in many environments. This study investigated the response of housefly (Musca domestica) head AChE (HF-AChE) exposed to five OPs; chlorpyrifos (CPF), malathion (MLT), triazophos (TRZ), monocrotophos (MCP) and profenofos (PRF) and two CBs; carbaryl (CRB) and carbofuran (CBF) as individual compounds and as binary mixtures of OPs and CBs under in vitro conditions. In addition, the selected OPs and CBs were evaluated for their toxicity in binary combinations with two SPs; deltamethrin (DLT) and cypermethrin (CYP) at fixed concentrations of 0.1 and 10μg/L. The toxicological interaction of five OPs with two CBs pesticides was evaluated under oxidised and un-oxidised conditions using a toxic unit (TU) approach and a concentration addition (CA) model. Pyrethroid combinations were assessed only under oxidised conditions. Since OPs and CBs act by a similar mechanism of inhibition of AChE, a dose additive effect was expected, but not conclusively found. TRZ with either CBF or CRB exhibited synergism under oxidised and un-oxidised conditions but the degree of synergism was stronger under un-oxidised conditions. Additivity was exhibited by CBF+MCP, CRB+MCP, CRB+MLT and CBF+MCP under un-oxidised conditions and CRB+MCP and CRB+CPF under oxidised conditions. Pyrethorids in combination with OPs (TRZ, MLT and CPF) were highly synergistic. In the present study, we used pure housefly head AChE without any interference of monooxygenase and/or esterase enzyme activities. Therefore these other enzymes were not producing the observed deviations from concentration-addition in the binary combinations between OPs, CBs and SPs. The mechanisms of OP, CB and SP interactions in pesticide mixtures requires further investigation.

Novel and selective acetylcholinesterase inhibitors for Tetranychus cinnabarinus (Acari: Tetranychidae)

The carmine spider mite, Tetranychus cinnabarinus (Acari: Tetranychidae), is an economically important and extremely polyphagous herbivorous pest, with the title of “resistance champion” among arthropods. Anticholinesterase insecticides such as organophosphate and carbamate account for more than one-third of global insecticide sales. The non-target toxicity and resistance problem of organophosphate and carbamate have become of growing concern, which may be due to the fact that they target the ubiquitous catalytic serine residue of acetylcholinesterase (AChE) in mammals, birds, and beneficial insects. In this study, the structural differences between T. cinnabarinus AChE and human AChE, at or near the catalytic pocket, were illustrated. From the SPECS chemical lead-compound database, 55 AChE inhibitor candidates were screened for high affinity for T. cinnabarinus AChE, but low affinity for human AChE, using the DOCK 6 and AutoDock Vina software. Three of the fifty-five candidates had inhibitory activity greater than that of the reversible AChE inhibitor eserine, with no observed inhibitory activities against human AChE. Two of the three had toxicity to T. cinnabarinus comparable to that of natural insecticidal pyrethrins. However, their potency is low compared with that of etoxazole, and further work is needed to optimize their potency. The selectivity of the three compounds over human and mite AChE may be due to their interaction with the mite-specific residues, as analyzed by Cyscore. The three compounds are potential lead compounds for development of novel acaricides against T. cinnabarinus with reduced toxicity to non-target species and a low propensity for resistance.