Toxicity Of Fipronil Research Articles

Phyto-toxicological effect of fipronil to plant seedlings: Assessing germination attributes, root-tip morphology, oxidative stress, and cellular respiration indices

Pesticides including insecticides are applied in agricultural practices to control insect pests. However, their excessive usage often poses a severe threat to the growth, physiology, and biochemistry of plants. Here, responses of chickpea and greengram seedlings exposed to three fipronil (FIP) concentrations i. e. 100 (1×), 200 (2×) and 300 (3×) μg mL−1 was evaluated under in vitro. Among doses, 3× had a greater negative impact on germination attributes, root-shoot elongation, vigor indices, length ratios, and survival of seedlings. Besides, the morphological distortion in root tips, oxidative stress generation, and cellular death in fipronil-supplemented root seedlings were observed under scanning electron (SEM) and confocal laser scanning (CLSM), respectively. A significant (p ≤ 0.05) and pronounced upsurge in plant stressor metabolites such as proline, malondialdehyde (MDA), electrolyte leakage (EL), hydrogen peroxide (H2O2) content, and antioxidants enzymes in plant seedlings further confirmed the fipronil toxicity. In addition, a concentration-dependent decrease in respiration efficiency (RE) and ATP content in FIP-treated seedlings was observed. Reduced mitotic index (MI) and numerous chromosomal anomalies (CAs) in root meristematic cells of seedlings are a clear indication of insecticide-induced cytotoxicity. Furthermore, a dose-dependent increase in DNA damage in root meristematic cells of greengram revealed the genotoxic potential of fipronil. Conclusively, fipronil suggested phyto and cyto-genotoxic effects that emphasize their careful monitoring in soils before application and their optimum addition in soil-plant systems. It is high time to prepare both target-specific and slow-released agrochemical formulations for crop protection with concurrent safeguarding of the soil.

Influences of TiO2 nanoparticle and fipronil co-exposure on metabolite profiles in mouse intestines.

Food contaminates, such as insecticide, may influence the toxicity of nanoparticles (NPs) to intestine. The present study investigated the combined toxicity of TiO2 NPs and fipronil to male mouse intestine. Juvenile mice (8weeks) were orally exposed to 5.74 mg/kg TiO2 NPs, 2.5mg/kg fipronil, or both, once a day, for 5days. We found that both TiO2 NPs and fipronil induced some pathological changes in intestines, accompanying with defective autophagy, but these effects were not obviously enhanced after TiO2 NP and fipronil co-exposure. Fipronil promoted Ti accumulation but induced minimal impact on other trace elements in TiO2 NP-exposed intestines. Metabolomics data revealed that the exposure altered metabolite profiles in mouse intestines, and two KEGG pathways, namely, ascorbate and aldarate metabolism (mmu00053) and glutathione metabolism (mmu00480), were only statistically significantly changed after TiO2 NP and fipronil co-exposure. Five metabolites, including 2-deoxy-D-erythro-pentofuranose 5-phosphate, 5alpha-cholestanol, beta-D-glucopyranuronic acid, elaidic acid, and isopentadecanoic acid, and maltotriose, were more significantly up-regulated after the co-exposure, whereas trisaccharide and xylonolactone were only significantly down-regulated by the co-exposure. We concluded that fipronil had minimal impact to enhance the toxicity of TiO2 NPs to mouse intestines but altered metabolite profiles.

Study of Protective Effect of Curcumin and Quercetin Against Subacute Toxicity of Fipronil in Rats

Study of Protective Effect of Curcumin and Quercetin Against Subacute Toxicity of Fipronil in Rats

Temperature influences the toxicity, behavioral, and physiology response of chlorfenapyr, fipronil, imidacloprid, and indoxacarb to Reticulitermes flaviceps

A comprehensive understanding of the interaction between pesticides and climate change is critical to the safety of pesticide applications. Many insecticides are actually more toxic at higher temperatures. However, little is currently known about the effect of temperature on the toxicity of chemicals. In order to fill this knowledge gap and improve our understanding of the mechanisms of temperature effects, the present study investigated the toxic, physiological, and behavioral effects of four insecticides, chlorfenapyr, fipronil, imidacloprid, and indoxacarb, on subterranean termites, Reticulitermes flaviceps at different temperatures (24 to 32 °C). The results show that the sensitivity of R. flaviceps to chrofenapyr (LC50 = 8.67–152.49 mg/L), fipronil (LC50 = 10.14–36.84 mg/L), imidacloprid (LC50 = 184.27–326.47 mg/L) and indoxacarb (LC50 = 12.23–46.23 mg/L) substantially increased when the temperature alters from 24 to 32 °C. Additionally, the activities of three detoxification enzymes [glutathione S-transferases (GSTs), carboxylesterase (CarE), and cytochrome P450 (P450)] of R. flaviceps were also measured of four insecticide at various temperatures. Correlation analysis reveals that changes in GSTs CarE and P450 activities of R. flaviceps are closely related to their sensitivity to chlorfenapyr, fipronil, imidacloprid, and indoxacarb. Meanwhile, the speed of action (LT50 value) of four insecticides is positively affected by temperature. In addition, intoxication, ataxia, and moribundity behaviors were observed in most concentrations and temperature combinations for chlorfenapyr, fipronil, and imidacloprid–exposed termite. The findings of our research emphasize that in the context of global climate change, the development and application of pesticides should consider the temperature dimension.

Ecological consequences when organisms avoid a contaminated environment: A study evaluating the toxicity of fipronil

The ability of aquatic organisms to sense the surrounding environment chemically and interpret these signals correctly is crucial to their survival and ecological niche. This study applied the Heterogenous Multi-Habitat Assay System - HeMHAS to evaluate the avoidance potential of Daphnia magna to detect fipronil-contaminated habitats in a connected landscape after a short (48 h), previous, forced exposure to an environmentally relevant concentration of the same insecticide. The swimming of daphnids was also analyzed by recording the total distance covered. D. magna preferred areas with less contamination, although the effect of fipronil on their swimming ability (a decrease) was observed for all the concentrations tested. The application of non-forced multi-compartment exposure methodologies is a recent trend and is ecologically relevant as it is based on how contamination can really produce changes in an organism's habitat selection. Finally, we consider the importance of more non-forced exposure approaches where Stress Ecology can be aggregated to improve systemic understanding of the risk that contaminants pose to aquatic ecosystems from a broader landscape perspective.

Assessment of Fipronil toxicity following subacute dermal exposure in rats

Assessment of Fipronil toxicity following subacute dermal exposure in rats

Fipronil Toxicity Induced Hemato-Biochemical Alterations in Male Wistar Albino Rats and its Alleviation with Pomegranate Peel Extract (Punica granatum)

Fipronil Toxicity Induced Hemato-Biochemical Alterations in Male Wistar Albino Rats and its Alleviation with Pomegranate Peel Extract (Punica granatum)

Effect of Fipronil Exposure on Hematological Aspects of Rhesus Monkeys (Macaca mulatta): Risk and Toxicity Assessment in Agro-Workers.

Fipronil (FPN) is a broad-spectrum phenylpyrazole insecticide, widely used in agriculture and veterinary medicine. Published research on FPN toxicity has established the fact that its inhalation or dermal exposure may lead to very serious clinical outcomes in non-target animals. In line to its exposure and toxicity related damage, FPN has been investigated in many invertebrates, however, its exposure-related noxiousness is less reported in higher animals. To assess the FPN-induced effects to agro-workers in the field, in the present study, we used physiological human surrogates, adult rhesus monkeys as models. We exposed well habituated, chair restraint adult rhesus monkeys with a field spray concentration of FPN (0.3 mg/1 mL distilled water) through an inhalation route in the closed system. Animals were divided into control and treatment groups, each containing three animals. Inflammatory and hematological effects were determined by evaluating the kidney and liver biomarker enzymes; serum creatinine and alanine transaminase (ALT), aspartate transaminase (AST) levels respectively. Our findings reveal that FPN treated monkeys show significantly increased levels of ALT (p = 0.000461), AST (p = 0.0681) and creatinine (p = 0.00656) as compared to the control group. Furthermore, significant differences of red blood cells (RBCs) (p = 0.0139) and white blood cells (WBCs) (p = 0.00642) were also observed in the treated and control group monkeys which reflect strong toxic effects on the blood cells. Our findings demonstrate that FPN exposure is very toxic to higher animals and causes severe damage to the liver and kidneys along with other clinical problems. The study highlights the effect and impact of passive inhalation of insecticides in intentionally carefree agro-workers and raises the concern of public awareness toward pesticides use.

Contact toxicity and proximate effect of fipronil on insect pest and predatory ant community structure in cocoa agro-ecosystem

Although usage persists in some countries, fipronil is banned or restricted in many others. Prior to its ban on cocoa in Ghana, concerns about its effect on non-target insects and secondary outbreak of Anomis leona were conflicting. This study, which predates the ban, assessed the toxicity and the short-term effect of fipronil on specific insect community structure in the cocoa agro-ecosystem alongside bifenthrin and a non-insecticide control. Although the insecticides induced a high mortality (90-100%) on the target (mirid: Sahlbergella singularis, stink bug: Bathycoelia thalassina and coreid bug: Pseudotheraptus devastans) and non-target (ants: Oecophylla longinoda, Crematogaster africana, Pheidole megacephala and Camponotus consobrinus) insects, the knockdown to fipronil was very low compared to bifenthrin. On the field, fipronil was more detrimental to the ants. Insecticide-treated plots recorded relatively lower post-treatment pest diversity compared to the control, except the last sampling month while ant abundance, richness and diversity were lowest on the fipronil-treated plots at the end of the study period. This study demonstrates that although fipronil was effective against pests and did not result in acute secondary pest outbreak, it was harmful to the ants. This effect could potentially be replicated on these ant species in other cropping systems where the insecticide is used, adversely affecting ecosystem service delivery. Hence, research on its impact on non-target organisms in other cropping systems is needed to regulate and monitor its use.

Toxicity assessment of carvacrol and its acetylated derivative in early staged zebrafish (Danio rerio): Safer alternatives to fipronil-based pesticides?

Fipronil is a broad-spectrum pesticide presenting high acute toxicity to non-target organisms, particularly to aquatic species. Natural compounds stand out as promising alternatives to the use of synthetic pesticides such as fipronil. Thus, our study aimed to compare the toxicity of carvacrol (natural), acetylcarvacrol (semisynthetic), and fipronil (synthetic) to early staged zebrafish. We conducted a series of toxicity assays at concentrations ranging from 0.01 μM to 25 μM for fipronil and 0.01 μM to 200 μM for carvacrol and acetylcarvacrol, depending on the assay, after 7-days post-fertilization (dpf). The potency (EC50) of fipronil was ∼1 μM for both deformities and mortality at 7 dpf, whereas EC50 was >50 μM for carvacrol and >70 μM for acetylcarvacrol. Fipronil at 0.1 and 1 μM caused a decrease in body length and swim bladder area of larvae at 7dpf, but no difference was observed for either carvacrol or acetylcarvacrol. Based upon the visual motor response test, fipronil induced hypoactivity in larval zebrafish at 1 μM and acetylcarvacrol induced hyperactivity at 0.1 μM. Anxiolytic-type behaviors were not affected by any of these chemicals. All chemicals increased the production of reactive oxygen species at 7 dpf, but not at 2 dpf. Genes related to swim bladder inflation, oxidative stress, lipid metabolism, and mitochondrial activity were measured; only fipronil induced upregulation of atp5f1c. There were no changes were observed in oxygen consumption rates of fish and apoptosis. Taken together, our data suggest that carvacrol and its derivative may be safer replacements for fipronil due to their lower acute toxicity.

Protective Effect of Uncaria Tomentosa Extract Against Induced Toxicity of Fipronil on Oral Mucosa of Rats. (Histological and Immunohistochemical study).

Protective Effect of Uncaria Tomentosa Extract Against Induced Toxicity of Fipronil on Oral Mucosa of Rats. (Histological and Immunohistochemical study).

Developmental toxicity of fipronil and its two metabolites towards zebrafish (Danio rerio) embryos

Fipronil (FIL) use is currently regulated in several countries due to its specific toxicity to bees. This study investigated the possible developmental toxicities as well as the acute toxicities of FIL, fipronil sulfide (FIL-SI), and fipronil sulfone (FIL-SO) to zebrafish (Danio rerio) embryos. At concentrations up to 5000 μg L−1, FIL- and FIL–SI–treated embryos exhibited significant mortality at 96 h postfertilization. Body length was significantly shortened with increasing concentrations in FIL- and FIL–SI–treated embryos. However, FIL–SO–treated embryos exhibited low mortality with high hatching rates. Body length was also significantly shortened in FIL–SO–treated embryos. Regarding the number of intersegmental vessels (ISVs), all chemical-treated embryos showed high ISV numbers with increasing concentrations of each chemical. FIL and FIL-SI induced abnormal heart formation with heart dysfunction in embryos, whereas FIL-SO did not induce any difference in heart development compared with the control. Abnormal heart formation may be related to the upregulation of nppa responsible for the expression of natriuretic peptides in embryos. Embryonic acetylcholinesterase activity was decreased gradually according to the increase in FIL and FIL-SI concentrations, whereas FIL-SO did not cause any change in enzyme activity. Il-1β responsible for the occurrence of injury or infection was highly upregulated in FIL–SI– and FIL–SO–treated embryos. Therefore, reduction to FIL-SI may be associated with FIL toxicity, whereas oxidation to FIL-SO may be a detoxification route in the environment.

Biochemical alterations in cotton leafworm, Spodoptera littoralis (Boisd.) related to emamectin benzoate and fipronil compared to their joint action

Cotton leafworm, Spodoptera littoralis (Boisduval), is one of the major destructive pests of ornamental, industrial, and vegetable crops. The efficacy of technical emamectin benzoate (EMB) and fipronil (FPR) was assessed against the 4th larval instar using leaf-dip bioassay method. EMB was more efficient than FPR based on 96 h LC50 values of 0.004 and 0.023 μg/ml, respectively. Joint toxic action of the dual exposure in sequence with time interval 24 h and in mix were evaluated at LC10:LC10, LC25:LC25 and LC50:LC50 after 96 h posttreatment, as well. Their impacts on detoxification enzymes, esterases (ESTs); alkaline phosphatase (ALP); and glutathione S-transferase (GST) as well as acetylcholine esterase (AChE) were also determined. The sequential exposure of EMB after FPR (S1) produced antagonism, potentiation, and potentiation effects, respectively while sequential exposure of FPR after EMB (S2) interacted as addition, potentiation, and potentiation respectively. The rest of binary mixtures (Mix) revealed antagonistic effect regardless of concentration. Orthogonal contrast analysis showed that the highest elevations of AChE, α-EST, β- EST and ALP enzymes were obtained from Mix at LC50:LC50 (181.6%, 288.4, 229.2 and 460.9%, respectively), LC25:LC25 (131.5%, 252.8, 205.60 and 252.0, respectively) and LC10:LC10 (106.6%, 215.6%, 201.8% and 170.0%, respectively). Differently, the greatest elevation of GST activity (157.7%) resulted from S1 at LC50:LC50, while it was significantly lower at LC25:LC25 and LC10:LC10 as well as Mix and S2 at all concentrations than corresponding concentrations of FPR. These findings shed some light on the role of GST in FPR toxicity and clarified the risk of these dual exposures in elevating detoxification enzymes dangerously compared to their individual insecticides. These dual exposures should be carefully handled. Although rotational exposure at low concentrations may enhance performance and mitigate resistance risk, rotational exposure at high concentrations and Mix may indirectly contribute to the evolution of cross-resistance to other insecticides.

Effect of melatonin and luzindole antagonist on fipronil toxicity, detoxification and antioxidant enzyme system in different tissues of Helicoverpa armigera (Lepidoptera: Noctuidae)

Studies have investigating the detoxification and antioxidant enzymes with melatonin under pesticide stress in many vertebrates, whereas no reports produced in invertebrates. In this study possible role of melatonin and luzindole effect on fipronil toxicity and the detoxification, antioxidant enzymes in H. armigera has been reported. Result showed high toxicity of fipronil treatment (LC50 4.24 ppm), followed by increased LC50 value with melatonin pretreatment (6.44 ppm). Whereas decreased toxicity was observed with melatonin and luzindole combination (3.72 ppm). The detoxification enzymes AChE, esterase and P450 were increased in larval head and whole body with exogenous melatonin level compared to control 1–1.5 μmol/mg of protein. The antioxidant levels of CAT, SOD and GST in whole body and head tissue had been increased by melatonin and fipronil combination 1.1–1.4 unit/mg of protein followed by GPx and GR in larval head (1–1.2 μmol/mg of protein). Mean while the luzindole antagonist inhibits CAT, SOD, GST and GR oxidative enzyme level (1–1.5 fold) in most of the tissue compared to melatonin and fipronil treatment (p < 0.01). Hence this study concludes that the melatonin pretreatment can reduce the fipronil toxicity by enhanced detoxification and antioxidant enzyme system in H. armigera.

Toxicity of fipronil and 2,4-D pesticides in Daphnia similis: a multiple endpoint approach

In Brazil, among the pesticides widely applied simultaneously in sugarcane monocultures are the Regent® 800 WG insecticide (active ingredient (a.i.) fipronil) and the DMA® 806 BR herbicide (a.i. 2,4-D). Thus, this study aimed to investigate, through different endpoints, the effects of the fipronil and 2,4-D pesticides, isolated and as mixtures, on the cladoceran Daphnia similis. To do this, acute toxicity tests were carried out with the compounds acting in isolation and in mixture, where the survival of the organisms was evaluated, and chronic toxicity tests with the isolated compounds, where reproduction and maternal and neonatal body length were evaluated. In this study, the physiological endpoints of D. similis were also analyzed, through the analysis of feeding rates (filtration and ingestion) in exposure and post-exposure scenarios, in order to verify the cladoceran food recovery capacity. In addition, D. similis data were compared with other species when exposed to the studied pesticides, using species sensitivity distribution curves. Acute toxicity tests of the fipronil and 2,4-D showed an average EC50-48h of 66.68μg a.i./L and 327.07mg a.i./L, respectively. In both cases, D. similis showed lower sensitivity compared to other species. For the mixture test, the evaluation by the IA model (independent action) and deviation DR (dose ratio dependent) indicated the occurrence of mostly antagonistic effects. The chronic test with fipronil showed a decrease in the fecundity of the organism at a concentration of 16μg a.i./L, a concentration already found in aquatic environments. For 2,4-D, no significant differences were observed for reproduction at the concentrations tested. Regarding the maternal body length, there were no significant changes when D. similis were exposed to both fipronil and 2,4-D, but these differences were observed in the body length of the neonates only for 2,4-D. There were no significant changes in the feeding rates of the organisms when exposed to both pesticides.

Individual and combined toxicity of imidacloprid and two seed dressing insecticides on collembolans Folsomia candida

ABSTRACT The aim of this study was to examine the chronic toxicity of imidacloprid (IMI), clothianidin (CLO) and fipronil (FIP) as a single exposure, as well as binary mixtures of IMI with CLO or FIP toward collembolans Folsomia candida, which are fauna present in the soil. Chronic toxicity assays were performed following an ISO guideline in a Tropical Artificial Soil (TAS), and the influence on the number and growth of the juveniles produced were determined. The range of nominal concentrations used in the tests with the individual compounds was 0.08–1.28 mg/kg (IMI), 0.079–1.264 mg/kg (FIP) and 0.007–0.112 mg/kg (CLO), whereas the mixture assays were performed with half the value used in the tests with individual compounds. Based upon single exposures, IMI produced a similar impact of reducing reproduction by 50% (EC50 ranging from 0.74 to 0.85 mg/kg) compared to FIP (EC50 = 0.78 mg/kg), whereas CLO was the most toxic to F. candida (EC50 = 0.08 mg/kg). Their mixtures generally resulted in a diminished effect on reproduction, as evidenced by the higher EC50 values. In contrast, in the case of the IMI+FIP combination at high concentrations at the EC50 level, a synergistic effect on toxicity was observed. The single exposure to the three insecticides and the mixture of IMI-FIP also decreased the size of generated juveniles, which was evidenced by the reduction in the proportion of large juveniles and increased proportion of small juveniles. However, both binary mixtures (IMI-FIP and IMI-CLO) presented antagonistic effects as evidenced by less than expected reductions in growth. Data on the toxic effects of IMI in a mixture with other seed dressing insecticides to collembolans provides useful information to environmental risk assessors by diminishing the uncertainties on the ecological risk of exposure to pesticides, enabling soil management degradation by utilizing multiple insecticides.

Dietary supplementation with Moringa oleifera leaves extract reduces the impacts of sub-lethal fipronil in Nile tilapia, Oreochromis niloticus

This study assessed the restorative dietary effects of Moringa oleifera (MO) leaves extract against the negative impacts of sub-lethal fipronil (FIP) toxicity in Nile tilapia. To achieve this purpose, the growth, body composition, haemato-biochemical measurements, serum immunity, and antioxidant condition of Nile tilapia have been examined. Fish were arranged into 6 experimental groups in quadruplicates. Three groups were fed on diets supplemented with 0.0 (reference group), 1.0 (MO1), and 2.0 (MO2) g kg−1 of MO leaf extract. The other three groups were fed on the same MO levels and concomitantly subjected to a sub-lethal FIP concentration (4.2 µg L−1 for 3 h only per day) and defined as FIP, FIP + MO1, and FIP + MO2. The experiment lasted for 8 weeks. Results unveiled that growth parameters were significantly decreased alongside an increased feed conversion ratio in the FIP-intoxicated group. The moisture and crude protein (%) were decreased significantly together with a significant increase of the crude lipids (%) in the fish body of the FIP group. Sub-lethal FIP toxicity induced hypochromic anemia, leukopenia, hypoproteinemia, hypoalbuminemia, hypoglobulinemia, and hepato-renal failure (increased urea and creatinine concentrations, as well as ALT and AST enzymes). Exposure to sub-lethal FIP also induced (a) immunosuppression manifested by a decline in total IgM, complement C3, and lysozyme activities, (b) enzymatic antioxidant misbalance manifested by decreases in SOD and CAT activities, and (c) oxidative stress (declined T-AOC and elevated of MDA concentrations). On the other side, dietary supplementation with MO leaf extract in FIP + MO1 and FIP + MO2 groups noticeably modulated the aforementioned parameters. Therefore, we can conclude that dietary MO could reduce sub-lethal FIP toxicity in Nile tilapia with a possible recommendation for regular prophylaxis supplementation in Nile tilapia diets.

Insights into the toxicity and biodegradation of fipronil in contaminated environment

Fipronil is a phenylpyrazole insecticide used in various agricultural, horticulture, and veterinary practices. Besides its wide range of applications, it also causes severe health hazards to the non-targeted organisms especially in developing countries. Fipronil showed hepatotoxic, nephrotoxic, neurotoxic, and altered reproductive development and endocrine system in humans and animals. Several methods have been already introduced for the removal of toxic fipronil including physicochemical and by the implementation of microorganisms. The microbial methods of fipronil degradation are the most promising and environmentally sustainable. The remediation of fipronil from the environment is an emerging task due to its enhanced residual concentration. Herein, we discuss the bioremediation potential of microbial strains in contaminated soil and water. It is shown that fipronil can be remediated from the environment using combined ecotechnologies. This review discusses the toxicity, different physico-chemical and biological methods, and sustainable developments in fipronil-contaminated agriculture and aquatic environments.

Influence of dissolved organic carbon on multimedia distribution and toxicity of fipronil and its transformation products in lotic waterways

Environmental fate and ecological impacts of fipronil and its transformation products (FIPs) in aquatic environment have caused worldwide attention, however, the influence of dissolved organic carbon (DOC) on multimedia distribution, bioavailability, and toxicity of FIPs in field waterways was largely unknown. Here, we collected 11 companion water and sediment samples along a lotic stream in Guangzhou, South China. FIPs were ubiquitous with total water concentrations ranging from 1.22 to 43.2 ng/L (14.8 ± 12.9 ng/L) and fipronil sulfone was predominant in both water and sediment. More than 70% of FIPs in aqueous phase were bound to DOC and the KDOC values of FIPs were approximately 1–2 orders of magnitude higher than Kd-s/KOC, emphasizing the significance of DOC in phase partitioning and transport of FIPs in aquatic environment. Water and sediment samples were more toxic to Chironomus dilutus than Hyallela azteca, and FIPs (especially fipronil sulfone) pronouncedly contributed toxicity to C. dilutus. Toxic units (TU) based on freely dissolved concentrations in water determined by solid phase microextraction significantly improved toxicity estimation of FIPs to the invertebrates compared to TUs based on aqueous concentrations. The present study highlights the significance of DOC association on fate and ecological risk of hydrophobic insecticides in lotic ecosystem.

Muscarinic acetylcholine receptor activation synergizes the knockdown and toxicity of GABA-gated chloride channel insecticides.

Pest management requires continual identification of new physiological targets and strategies to control pests affecting agriculture and public/animal health. We propose the muscarinic system as a target for agrochemicals because of its physiological importance. Unlike the muscarinic system, gamma-amino butyric acid (GABA) receptors are an established insecticide target. Here, we investigated target-site synergism using small molecule probes (agonist and antagonist) against the muscarinic system and their ability to enhance the toxicity of GABAergic insecticides in Drosophila melanogaster (Meigen). Oral delivery of pilocarpine (muscarinic agonist) enhanced the toxicity of dieldrin, fipronil, and lindane, resulting in synergist ratios (SRs) between 4-32-fold (orally delivered) or between 2-67-fold when insecticides were topically applied. The synergism between pilocarpine and the GABA-insecticides was greater than the synergism observed with atropine (muscarinic antagonist), and was greater, or comparable, to the synergism observed with the metabolic inhibitor piperonyl butoxide. In addition to lethality, pilocarpine increased the knockdown of lindane. The mechanism of synergism was also investigated in the central nervous system using extracellular electrophysiology, where pilocarpine (3μmo/L) lowered the half-maximal inhibitory concentration (IC50 ) of lindane from 1.3 (0.86-1.98) μmol/L to 0.17 (0.14-0.21) μmol/L and fipronil's IC50 from 2.2 (1.54-3.29) μmol/L to 0.56 (0.40-0.77) μmol/L. Convergence of the cellular function between the muscarinic and GABAergic systems enhanced the insecticidal activity of GABA receptor blocking insecticides through the modulation of the central nervous system (CNS). The future impact of the findings could be the reduction of the active ingredient needed in a formulation with the development of muscarinic synergists. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.