Selective and sensitive molecularly imprinted polymer-based electrochemical sensor for detection of deltamethrin.

Electrochemical sensors have a broad range of industrial applications due to their sensitivity, speed, and cost-effectiveness. These sensors enable the continuous monitoring and control of critical parameters in various industrial processes. For instance, they are essential in food safety, environmental monitoring, biomedical applications, and pharmaceutical production. In the food industry, electrochemical sensors facilitate the rapid and reliable detection of contaminants and pathogens in food products, thus enhancing product quality and consumer safety. An electrochemical sensor was developed with the molecularly imprinted polymer (MIP) technique to detect deltamethrin with high sensitivity and selectivity. The sensor was fabricated by electrodeposition of Co3O4 on indium tin oxide (ITO), followed by electropolymerization of o-phenylenediamine with deltamethrin as a template molecule. The template molecules were then removed from the modified electrode by a methanol. The MIP-based electrochemical sensor exhibited high sensitivity and selectivity towards deltamethrin. Under the optimized conditions, the LOD values for the MIP/Co3O4/ITO electrode in the first and second linear regressions were 1.53 nM for linear range of 2.82 nM to 56.5 nM and 0.34 µM for linear range of 0.25 µM to 3.98 µM. Moreover, the LOD values for the NIP/Co3O4/ITO electrode in the first and second regressions were 2.43 nM for the linear range of 3.91 nM to 65.0 nM and 726.0 nM for the linear range of 0.023 µM to 4.5 µM. The developed electrochromic pesticide sensor, being an electrochemical-based molecularly imprinted polymer (MIP) sensor incorporating electrochromic materials, enables both target-specific pesticide detection and visual pesticide identification based on color changes dependent on pesticide concentration. Consequently, this system is more advantageous compared to electrochemical-based MIP sensors, as it provides both qualitative and quantitative determinations. The qualitative assessment aims to enhance the ease of use of the sensor, thereby increasing the potential for it to become a commercially viable product by reducing the need for instrumental devices.

Efficacy of PermaNet® Dual compared to Interceptor® G2 and PermaNet 3.0 in experimental huts in Siaya County, western Kenya.

BACKGROUND: Pyrethroid-chlorfenapyr nets have shown significant epidemiological impact over pyrethroid-only and pyrethroid plus piperonyl-butoxide (PBO) in Africa. A non-inferiority evaluation of PermaNet® Dual, a new chlorfenapyr plus deltamethrin net, compared to Interceptor® G2, was conducted in experimental huts in Siaya, Kenya against free-flying pyrethroid-resistant Anopheles funestus. METHODS: This study was an experimental hut trial, following a 7 by 7 Latin Square design. Seven treatments and seven sleepers were deployed in the experimental huts daily and rotated weekly and daily, respectively. Mosquitoes were collected every morning between 06:30 h and 08:30 h and were assessed for blood feeding and then monitored for immediate knockdown 1-h post collection and delayed mortality after 72 h. Differences in proportional outcomes were analysed using the blocked logistic regression model, while differences in numerical outcomes were analysed using the negative binomial regression model. Non-inferiority determination was performed based on World Health Organization (WHO) protocol. RESULTS: Mortality at 72 h was 30.2% for PermaNet 3.0, 44.4% for the Interceptor® G2 and 49.2% for the PermaNet® Dual. Blood feeding was highest with PermaNet® Dual at 15%, and least with PermaNet® 3.0 at 10%. PermaNet® Dual and Interceptor® G2 had no significant differences in mortality (OR = 1.10, 95% CI 1.00-1.20) or blood feeding (OR = 1.18, 95% CI 1.04-1.33) and the lower confidence bounds were within the non-inferiority margins but for blood feeding, non-inferiority was relatively high to the upper 95% confidence bound. PermaNet® Dual was non-inferior to the Interceptor® G2 and superior to the PermaNet® 3.0 nets in causing mortality but inferior to PermaNet ®3.0 in blood feeding inhibition of the vectors. CONCLUSION: PermaNet® Dual met the WHO criteria for non-inferiority to Interceptor® G2 and may be considered for deployment for public health use against pyrethroid-resistant Anopheles vectors of malaria.

Phenotypic evidence of deltamethrin resistance and identification of selective sweeps in Anopheles mosquitoes on the Bijagós Archipelago, Guinea-Bissau.

Vector control in the Bijagós Archipelago of Guinea-Bissau currently relies on pyrethroid insecticide-treated nets. However, data on insecticide resistance in Guinea-Bissau is limited. This study identified deltamethrin resistance in the Anopheles gambiae sensu lato complex on Bubaque island using WHO tube tests in November 2022. Whole genome sequencing of An. gambiae sensu stricto mosquitoes identified six single nucleotide polymorphisms (SNPs) previously associated with, or putatively associated with, insecticide resistance: T791M, L995F, N1570Y, A1746S and P1874L in the vgsc gene, and L119V in the gste2 gene. Twenty additional non-synonymous SNPs were identified in insecticide-resistance associated genes. Four of these SNPs were present at frequencies over 5% in the population: T154S, I126F and G26S in the vgsc gene and A65S in ace1. Genome wide selection scans using Garud's H12 statistic identified two selective sweeps: one in chromosome X and one in chromosome 2R. Both selective sweeps overlap with metabolic genes previously associated with insecticide resistance, including cyp9k1 and the cyp6aa/cyp6p gene cluster. This study presents the first phenotypic testing for deltamethrin resistance and the first whole genome sequence data for Anophelesgambiae mosquitoes from the Bijagós, contributing data of significance for vector control policy in this region.

The non-inferiority of piperonyl-butoxide Yorkool® G3 insecticide-treated nets compared to Olyset®Plus measured by Anopheles arabiensis mortality in experimental huts in Tanzania.

BACKGROUND: Non-inferiority trials are recommended by the World Health Organization (WHO) to demonstrate that health products show comparable efficacy to that of existing standard of care. As part of the WHO Global Malaria Programme (GMP) process of assessment of malaria vector control products, a second-in-class insecticide-treated net (ITN) must be shown to be non-inferior to a first-in-class product based on mosquito mortality. The public health impact of the first-in-class pyrethroid-piperonyl butoxide (PBO) ITN, Olyset® Plus, has been demonstrated in epidemiological trials in areas with insecticide-resistant mosquitoes, but there is a need to determine the efficacy of other pyrethroid-PBO nets to ensure timely market availability of nets in order to increase access to ITNs. The non-inferiority of a deltamethrin-PBO ITN Yorkool® G3 was evaluated entomologically against Olyset® Plus in experimental huts in Tanzania, following WHO guidelines for non-inferiority trials. METHODS: The trial of the two pyrethroid-PBO ITNs was conducted in experimental huts in Lupiro, Tanzania, using a randomized 7 × 7 Latin square block design. The study ran for 49 nights in 14 huts assessing the mosquito mortality and blood-feeding of wild, free-flying, pyrethroid-resistant Anopheles arabiensis. Using the non-inferiority approach, the comparative efficacy (primary endpoint was mosquito mortality at 24 h and secondary endpoint was blood-feeding) of unwashed and 20 times field-washed pyrethroid-PBO Yorkool® G3 ITNs, were compared with the first-in-class product Olyset® Plus and against a pyrethroid-only ITN, PermaNet® 2.0 ITNs, as a standard comparator. RESULTS: The experimental hut trial demonstrated non-inferiority and superiority of Yorkool® G3 to Olyset® Plus based on mosquito mortality [51% vs. 39%, OR 1.68 (95% CI 1.50-1.88)], given that lower 95% CI exceeded 0.74 (delta of 39%) and the margin of no difference (1). Blood-feeding inhibition was high for all treated ITNs (> 90%) and Yorkool® G3 was non-inferior to Olyset® Plus [4% vs. 2%, OR 1.81 (95% CI 1.46-2.39)], given that upper 95% CI was less than 4.85 (delta of 4%). The pyrethroid-PBO ITNs were superior to the pyrethroid-only net, PermaNet® 2.0, as determined by both the proportion of mortality and blood-feeding of mosquitoes (p-value < 0.05). CONCLUSION: Yorkool® G3 ITNs demonstrated non-inferiority to the first-in-class Olyset® Plus and superiority over the standard pyrethroid-only ITN, PermaNet® 2.0 as measured by mortality and blood-feeding inhibition of wild pyrethroid-resistant An. arabiensis mosquitoes. Yorkool® G3 ITNs are potential tools for the control of metabolic insecticide-resistant malaria vectors, and their market availability will contribute to the cost-effective selection of ITNs by malaria control programmes to improve population access to ITNs.

The mechanistic pathway induced by fenpropathrin toxicity: Oxidative stress, signaling pathway, and mitochondrial damage.

Fenpropathrin (FNP) is a kind of insecticide and acaricide known as pyrethroid. It is very effective, has a wide range of activities, and works quickly. Internationally, it is commonly considered the most powerful pyrethroid insecticide. Nevertheless, an increasing amount of data indicates a substantial link between Fenpropathrin and adverse effects on nontarget species, including liver toxicity, kidney toxicity, nerve damage, and reproductive toxicity. Oxidative stress plays a vital role in the toxicity of fenpropathrin, in addition to its mechanical mechanism. This study offers a thorough examination of the harmful effects of Fenpropathrin on oxidative and mitochondrial processes, as well as the signaling pathways involved in these effects. The significant impact of oxidative stress emphasizes the toxicity of Fenpropathrin.

Insecticide Resistance and Target-Site Mutations kdr, N1575Y, and Ace-1 in Anopheles gambiae s.l. Populations in a Low-Malaria-Transmission Zone in the Sudanian Region of Senegal.

Background/Objectives: Significant progress in malaria control has been achieved through long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS), raising hopes for malaria elimination. However, emerging insecticide resistance threatens these gains. This study assessed the susceptibility of Anopheles gambiae s.l. populations to public health insecticides, examined the frequencies of kdr, Ace-1, and N1575Y mutations, and explored their associations with phenotypic resistance in Dielmo and Ndiop, Senegal. Methods: Anopheles larvae collected between September and December 2022 were reared to adulthood. Adult mosquitoes were exposed to discriminating concentrations of various insecticides following WHO guidelines. Knockdown times (KDT50 and KDT95) for pyrethroids were calculated using the Probit model. RT-qPCR detected target-site mutations (kdr: L1014F and L1014S, Ace-1, N1575Y) and assessed correlations with phenotypic resistance. Species-specific PCR identified species within the An. gambiae complex. Results/Conclusions: The populations of Dielmo and Ndiop showed susceptibility to pirimiphos-methyl and bendiocarb, with no Ace-1 mutation detected. Resistance to DDT and pyrethroids was observed. The knockdown times indicated that alphacypermethrin and lambdacyhalothrin were more effective than permethrin and deltamethrin. The L1014F allele was widespread, while L1014S was absent in Ndiop and low in Dielmo. The N1575Y mutation occurred only in populations with L1014F. The L1014S mutation was significantly associated with resistance to lambdacyhalothrin in both villages and to deltamethrin in Ndiop.

An observational analysis of the impact of deltamethrin + piperonyl butoxide insecticide-treated nets on malaria case incidence and entomological indicators in Ebonyi State, Nigeria, 2017-2021.

BACKGROUND: Intense pyrethroid resistance threatens the effectiveness of the primary vector control intervention, insecticide-treated nets (ITNs), in Nigeria, the country with the largest malaria burden globally. In this study, the epidemiological and entomological impact of a new type of ITN (piperonyl-butoxide [PBO] ITNs) distributed in Ebonyi State were evaluated. The epidemiological impact was also compared to the impact of standard pyrethroid-only ITNs in Cross River State. METHODS: A controlled interrupted time series analysis was conducted on monthly malaria incidence data collected at the health facility level, using a multilevel mixed-effects negative binomial model. Data were analysed two years before and after the PBO ITN campaign in Ebonyi State (December 2017 to November 2021). A pre-post analysis, with no comparison group, was used to assess the impact of PBO ITNs on human biting rates and indoor resting density in Ebonyi during the high transmission season immediately before and after the PBO ITN campaign. RESULTS: In Ebonyi, PBO ITNs were associated with a 46.7% decrease (95%CI: -51.5, -40.8%; p < 0.001) in malaria case incidence in the 2 years after the PBO ITN distribution compared to a modelled scenario of no ITNs distributed, with a significant decrease from 269.6 predicted cases per 1000 population to 143.6. In Cross River, there was a significant 28.6% increase (95%CI: -10.4, 49.1%; p < 0.001) in malaria case incidence following the standard ITN distribution, with an increase from 71.2 predicted cases per 1000 population to 91.6. In Ebonyi, the human biting rate was 72% lower (IRR: 0.28; 95%CI 0.21, 0.39; p < 0.001) and indoor resting density was 73% lower (IRR: 0.27; 95%CI 0.21, 0.35; p < 0.001) after the PBO ITNs were distributed. CONCLUSIONS: The epidemiological and entomological impact of the PBO ITNs underscore the impact of these ITNs in areas with confirmed pyrethroid resistance. These findings contribute to ongoing research on the impact of new types of ITNs in Nigeria, providing critical evidence for the Nigeria National Malaria Elimination Programme and other countries for future ITN procurement decisions as part of mass ITN campaign planning and malaria programming.

The attrition, physical and insecticidal durability of two dual active ingredient nets (Interceptor® G2 and Royal Guard®) in Benin, West Africa: results from a durability study embedded in a cluster randomised controlled trial.

BACKGROUND: Studies evaluating the attrition, physical and insecticidal durability of dual active ingredient (AI) insecticide-treated nets (ITNs) are essential for making programmatic decisions regarding their deployment. We performed a prospective study embedded in a cluster randomised controlled trial (cRCT) to evaluate the attrition, fabric integrity and insecticidal durability of Interceptor® G2 (alpha-cypermethrin-chlorfenapyr) and Royal Guard® (alpha-cypermethrin-pyriproxyfen), compared to Interceptor® (alpha-cypermethrin) in Benin. METHODS: A total of 2428 study nets in 1093 randomly selected households in five clusters per arm of the cRCT were monitored for ITN attrition and fabric integrity every 6-12 months post-distribution. Householders were further surveyed to investigate non-study net use and their preference for ITN fabric types used in the study nets. A second cohort of 120 nets per ITN type were withdrawn every 12 months and assessed for chemical content and insecticidal activity in laboratory bioassays. Alpha-cypermethrin bioefficacy was investigated using the susceptible Anopheles gambiae Kisumu strain, and chlorfenapyr and pyriproxyfen bioefficacy were investigated using the pyrethroid-resistant Anopheles coluzzii Akron strain. Net pieces were tested in WHO cone bioassays and tunnel tests for alpha-cypermethrin and in tunnel tests for chlorfenapyr; pyriproxyfen activity was assessed in cone bioassays as the reduction in fertility of blood-fed survivors using ovary dissection. Bioefficacy was expressed as the proportion of ITNs passing predetermined WHO criteria, namely knock-down ≥ 95% or 24/72 h mortality ≥ 80% or reduction in fertility ≥ 50%. RESULTS: Overall ITN survivorship was 52% at 24 months and fell to 15% at 36 months. Median ITN survival time was lower with Royal Guard® relative to Interceptor® [1.6 vs 2.3 years; hazard ratio (HR) 1.49, 95% confidence interval (CI) 1.36-1.66; p < 0.001] and Interceptor® G2 (1.6 vs 2.1 years; HR 1.33, 95% CI 1.20-1.47; p < 0.001). Householders overwhelmingly preferred polyester nets over polyethylene nets (96%), and more Royal Guard® nets were replaced with spare polyester nets from previous campaigns. All Royal Guard® nets passed efficacy criteria for alpha-cypermethrin at all time points (100%) while ITN pass rates after 24 months had fallen to < 40% for pyriproxyfen and chlorfenapyr. The chemical content analysis showed a higher loss rate of the non-pyrethroid insecticides relative to the pyrethroids in each dual ingredient AI ITN; 74% vs 47% for Royal Guard® and 85% vs 63% for Interceptor® G2 at 36 months. CONCLUSIONS: The median ITN survival time for Interceptor® G2 (2.1 years) and Royal Guard® (1.6 years) in Benin is substantially lower than 3 years. Royal Guard® nets were discarded more quickly by householders, partly due to their low preference for polyethylene nets. The insecticidal activity of the non-pyrethroid insecticides in both dual AI ITNs was short-lived compared to alpha-cypermethrin. The results corroborate the findings from the cRCT conducted in Benin.

Malaria vector control in sub-Saharan Africa: complex trade-offs to combat the growing threat of insecticide resistance.

Mass distribution of insecticide-treated nets (ITNs) has been a key factor in reducing malaria cases and deaths in sub-Saharan Africa. A shortcoming has been the over-reliance on pyrethroid insecticides, with more than 2·13 billion pyrethroid ITNs (PY ITNs) distributed in the past two decades, leading to widespread pyrethroid resistance. Progressive changes are occurring, with increased deployment of more effective pyrethroid-chlorfenapyr (PY-CFP) or pyrethroid-piperonyl butoxide (PY-PBO) ITNs in areas of pyrethroid resistance. In 2023, PY-PBO ITNs accounted for 58% of all ITNs shipped to sub-Saharan Africa. PY-PBO and PY-CFP ITNs are 30-37% more expensive than standard PY ITNs, equating to an additional US$132-159 million required per year in sub-Saharan Africa to fund the shift to more effective ITNs. Several countries are withdrawing or scaling back indoor residual spraying (IRS) programmes to cover the shortfall, which is reflected by the number of structures sprayed by the US President's Malaria Initiative decreasing by 30% from 5·67 million (2021) to 3·96 million (2023). Benin, located in West Africa, is a prime example of a country that ceased IRS in 2021 after 14 years of annual spraying. Our economic evaluation indicates that IRS in Benin cost $3·50 per person protected per year, around five times more per person protected per year compared with PY-PBO ($0·73) or PY-CFP ITNs ($0·76). Although costly to implement, a major advantage of IRS is the portfolio of at least three chemical classes for prospective resistance management. With loss of synergy to PBO developing rapidly, there is the danger of over-reliance on PY-CFP ITNs. As gains in global malaria control continue to reverse each year, current WHO projections estimate that key 2030 malaria incidence milestones will be missed by a staggering 89%. This Personal View explores contemporary malaria vector control trends in sub-Saharan Africa and cost implications for improved disease control and resistance management.

Improving Sphenophorus levis Adult Mortality Through Solid Insecticide Applications and Increased Insecticide Dose.

The sugarcane weevil (Sphenophorus levis Vaurie, 1978) is currently considered the most important sugarcane pest in Brazil, causing significant yield losses. Application methods of insecticides for S. levis control have not been effective, mostly due to the insect's cryptic behavior below the soil surface which suppresses the correct placement of insecticide active ingredients on target. Two experiments were conducted using an innovative bioassay methodology that simulates sugarcane field conditions to effectively evaluate S. levis adult mortality and insecticide residues in the soil under different treatments. The first experiment aimed to assess the efficacy of two liquid- and solid-applied insecticides, while the second aimed to examine the effect of increasing the dose of lambda-cyhalothrin + thiamethoxam on S. levis adult control. The novel bioassays simulated liquid and solid insecticide applications on sugarcane by exposing S. levis adults to chemical residuals on rhizomes and in soil after insecticide application. In the first experiment, low S. levis adult control was detected (< 53% mortality) across all treatments, where both solid and liquid applications of lambda-cyhalothrin + thiamethoxam provided greater efficacy levels than imidacloprid and control treatments, respectively. Solid applications maintained higher insecticide concentrations in the soil for longer periods than liquid insecticide applications, providing maximum insect control levels 7 days after application. The second experiment revealed that solid applications at higher insecticide doses significantly improved control of S. levis adult (76.7% mortality) and resulted in greater insecticide concentrations in the soil compared to the recommended label rate (58.8% mortality).

A preliminary estimate of the environmental burden of disease associated with exposure to pyrethroid insecticides and ADHD in Europe based on human biomonitoring.

Human biomonitoring (HBM) data indicate that exposure to pyrethroids is widespread in Europe, with significantly higher exposure observed in children compared to adults. Epidemiological, toxicological, and mechanistic studies raise concerns for potential human health effects, particularly, behavioral effects such as attention deficit hyperactivity disorder (ADHD) in children at low levels of exposure. Based on an exposure-response function from a single European study and on available quality-assured and harmonized HBM data collected in France, Germany, Iceland, Switzerland, and Israel, a preliminary estimate of the environmental burden of disease for ADHD associated with pyrethroid exposure was made for individuals aged 0-19 years. The estimated annual number of prevalence-based disability-adjusted life years (DALYs) per million inhabitants were 27 DALYs for Israel, 21 DALYs for France, 12 DALYs for both Switzerland and Iceland, and 3 DALYs for Germany; while the annual ADHD cases per million inhabitants attributable to pyrethroids were 2189 for Israel, 1710 for France, 969 for Iceland, 944 for Switzerland, and 209 for Germany. Direct health costs related to ADHD ranged between 0.3 and 2.5 million EUR yearly per million inhabitants for the five countries. Additionally, a substantial number of ADHD cases, on average 18%, were associated with pyrethroid exposure. Yet, these figures should be interpreted with caution given the uncertainty of the estimation. A sensitivity analysis showed that by applying a different exposure-response function from outside the EU, the population attributable fraction decreased from an average of 18 to 7%. To ensure more robust disease burden estimates and adequate follow-up of policy measures, more HBM studies are needed, along with increased efforts to harmonize the design of epidemiological studies upfront to guarantee meta-analysis of exposure-response functions. This is particularly important for pyrethroids as evidence of potential adverse health effects is continuously emerging.

Cytochrome P450 CYP6AE70 Confers Resistance to Multiple Insecticides in a Lepidopteran Pest, Spodoptera exigua.

Cytochrome P450 monooxygenases are associated with the detoxification of xenobiotics, and overexpression of P450 genes has been proven to be associated with insecticide resistance in insect species. Our previous study has revealed that multiple CYP6AE genes were significantly overexpressed in a resistant strain of Spodoptera exigua, and among these genes, CYP6AE70 was particularly highly expressed. However, the functional roles of the CYP6AE genes in insecticide resistance remain unknown in this pest. Here, we investigate the relationship between the CYP6AE genes and insecticide resistance by focusing on CYP6AE70. The expression of CYP6AE70 was increased after exposure to chlorpyrifos, cypermethrin, and deltamethrin. Ectopic overexpression of P450 in transgenic flies by the GAL4/UAS system dramatically enhanced the tolerance to these three insecticides. Furthermore, the recombinant CYP6AE70 was functionally expressed in Sf9 cells, and metabolic assays revealed that the recombinant P450 protein could efficiently metabolize chlorpyrifos, cypermethrin, and deltamethrin. Finally, molecular modeling and docking also showed that this P450 protein were tightly bound to the three insecticides. These results determine that the upregulation of CYP6AE genes results in resistance to multiple insecticides in S. exigua. This study improves our understanding of P450-mediated insecticide resistance and will help us to design more effective resistance management for pest control.

Stability, Inheritance, Cross-Resistance, and Fitness Cost of Resistance to λ-Cyhalothrin in Cydia pomonella.

Insecticides are commonly utilized in agriculture and forestry for pest control, but their dispersal can pose hazards to humans and environment. Understanding resistance, inheritance patterns, and fitness costs can help manage resistance. A λ-cyhalothrin-resistant population (LCR) of Cydia pomonella, a global pest of pome fruits and walnuts, was obtained through selective insecticide breeding for 15 generations, showing stable moderate resistance (23.85-fold). This population was cross-resistant to deltamethrin (4.26-fold) but not to ß-cypermethrin, chlorantraniliprole, chlorpyrifos, and avermectin. Genetic analysis revealed the resistance was autosomal, incompletely dominant, and controlled by multiple genes. Increased activity of glutathione S-transferases and cytochrome P450 monooxygenases (P450s) played a primary role in resistance, with specific genes up-regulated in LCR, and exhibited significant expression in midgut. LCR also exhibited fitness costs, including delays in development, reduced fecundity, and slower population growth. These findings contribute to understanding λ-cyhalothrin resistance in C. pomonella and can guide resistance management strategies.

Airborne metofluthrin, a pyrethroid repellent, does not impact foraging honey bees.

Outdoor spatial mosquito repellents, such as mosquito coils or heating devices, release pyrethroid insecticides into the air to provide protection from mosquitoes within a defined area. This broadcast discharge of pyrethroids into the environment raises concern about the effect on non-target organisms. A previous study found that prallethrin discharged from a heating device did not affect honey bee (Apis mellifera L.) [Hymenoptera: Apidae] foraging or recruitment. In this second study, there was no significant difference in foraging frequency (our primary outcome), waggle dance propensity, or persistency in honey bees collecting sucrose solution between those exposed to metofluthrin from a different heating device and bees exposed to a non-metofluthrin control. One measure, waggle dance frequency, was higher in the metofluthrin treatment than the control but this outcome was likely a spurious result due to the small sample size. The small particle size of the emissions, averaging 4.43 µm, from the heated spatial repellent products, which remain airborne with little settling, may play an important role in the lack of effect found on honey bee foraging.

Toxicokinetic model of the pyrethroid pesticide lambda-cyhalothrin, main exposure route and dose reconstruction predictions in agricultural workers.

A toxicokinetic model of the pyrethroid insecticide lambda-cyhalothrin (LCT) was developed to relate absorbed doses to urinary cis-3-(2-chloro-3,3,3-trifluoroprop-1-en-1-yl)-2,2-dimethylcyclopropanecarboxylic acid (CFMP) metabolite levels used as a biomarker of exposure. The model then served to reconstruct absorbed doses in agricultural workers and their probability of exceeding the EFSA Acceptable occupational Exposure Level (AOEL). The toxicokinetic model was able to reproduce the temporal profiles of CFMP in the urine of operators spraying pesticides using the optimized model parameters (adjusted to human volunteer data). Modeling also showed that simulation of an inadvertent oral exposure mainly was the exposure scenario giving the best fit to CFMP urinary time-course data in applicators. With the dermal model parameters optimized from data in volunteers, simulation of a dermal exposure in applicators did not allow to reproduce the observed peak excretions and urinary metabolite levels; extremely high applied dermal doses would be required but still simulated dermal penetration rate would remain too slow. Simulation of an inhalation exposure allowed to reproduce the observed time-courses, but with unrealistic air concentrations. For applicators with the highest urinary concentrations, there was a probability of exceeding the AOEL at some points during the biomonitoring period [>50% probability of exceeding for 27% of 24-h samples]; for non-applicator workers the probability of exceeding the AOEL value was very low [corresponding value of 5%]. Furthermore, the median [95% CI] estimates of 10 000 Monte Carlo simulations led to a biological reference value corresponding to the AOEL of 116 [113-119] ng/kg bw/d and 7.5 [7.3-7.7] µg/L. Overall, 7% of applicators and 1% of workers performing weeding and strawberry picking had a probability of exceeding this biological reference value. As a next step, it would be interesting to apply these methods to multiple exposure to various contaminants.

Resistance of Anopheles gambiae s.s. against commonly used insecticides and implication of cytochrome P450 monooxygenase in resistance to pyrethroids in Lambaréné (Gabon).

BACKGROUND: Insecticides are a crucial component of vector control. However, resistance constitute a threat on their efficacy and the gains obtained over the years through malaria vector control. In Gabon, little data on phenotypic insecticide resistance in Anopheles vectors are published, compromising the rational implementation of resistance management strategies. We assessed the susceptibility to pyrethroids, carbamates and organophosphates of Anopheles gambiae sensu lato (s.l.) and discuss the mechanisms involved in the pyrethroid resistance-phenotype. METHODS: A. gambiae s.l. larvae were collected from breeding sites in Lambaréné. Emerging adults were used in WHO tube assays at an insecticide concentration that defines resistance (diagnostic concentration). Subsequently, deltamethrin and permethrin were used at 5x and 10x diagnostic concentrations and after preexposure with the cytochrome p450 (and glutathione S-transferase) inhibitor piperonyl butoxide (PBO). A subset of mosquitoes was typed by molecular methods and screened using Taqman assays for mutations conferring target site resistance at the Voltage-gated sodium channel 1014 (Vgsc-1014) locus and the acetylcholinesterase (Ace-1) gene. RESULTS: All mosquitoes were A. gambiae sensu stricto (s.s.) and resistant to permethrin, deltamethrin and alphacypermethrin (mortality less than 98%). However, mosquitoes were susceptible to malathion but resistant to bendiocarb. The level of resistance was high for permethrin and at least moderate for deltamethrin. Pre-exposure to PBO significantly increased the mortality of resistant mosquitoes (P < 0.0001). They became fully susceptible to deltamethrin and permethrin-induced mortality increased 4-fold. The G119S Ace-1 resistance allele, which confers resistance to both organophosphates and carbamates, was not present. All sampled mosquitoes were either homozygous for the Vgsc-L1014F or heterozygous for Vgsc-L1014F/L1014S, a marker for resistance to pyrethroids and organochlorides. CONCLUSION: These findings demonstrate a role of cytochrome P450 monooxygenases in the pyrethroid-resistance of A. gambiae s.s. from Lambaréné. Combining PBO with pyrethroids, as done in second generation bednets, may be used to revert resistance. In addition, malathion could also be used in combination with pyrethroids-based methods for resistance management.

Study on susceptibility differences of Tetranychus cinnabarinus (Boisduval) males and females to acaricides.

The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is an important agricultural arthropod pest. This study investigated acaricide susceptibility differences between sexes to determine the biochemical mechanisms potentially involved in these differences. The susceptibility of females and males to seven acaricides (pyridaben, cyflumetofen, abamectin, chlorpyrifos, propargite, profenofos and fenpropathrin) was compared with laboratory bioassays. Males were more susceptible than females for each acaricide. Differences in the lethal concentration 50 % (LC50) values between the sexes were observed to range from 2.04-fold to 6.05-fold. The surface area was markedly greater for females than males (by 1.87-fold) and the weight was significantly greater for females compared to males (by 4.67-fold). The specific surface area of male was 2.61-fold higher than that of female. There were no differences in the cuticle structure, whereas the thickness of females was 1.63-fold than that of males. The penetration rate of males was statistically significantly higher than that of females, both in pyridaben and cyflumetofen. Synergism experiments and biochemical assays suggested the involvement of the three detoxification enzyme systems in the sexual susceptibility of T. cinnabarinus. The activities of cytochrome P450 monooxygenases (P450s) (p < 0.01), glutathione-S-transferase (GSTs) (p < 0.05) and carboxylesterase (CarEs) (p < 0.05) in females were significantly higher than that in males. The results reveal that the individual size (specific surface area), cuticle thickness, and detoxification enzyme activity were involved in the sexual susceptibility to acaricides of T. cinnabarinus.

The down-regulation of salivary protein gene expression by etofenprox partially contributed to reducing the risk of increased fecundity in the brown planthopper.

Etofenprox is a pyrethroid insecticide that acts on the nervous system of insects. Due to its low toxicity to aquatic animals, it is permitted for use in controlling insect pests in rice fields. The brown planthopper (BPH), Nilaparvata lugens, a significant piercing-sucking pest feeding on rice exclusively, secretes various salivary components when feeding. Salivary proteins are essential for BPH feeding, but their response to etofenprox is not well understood. The application of etofenprox down-regulated the expression of 21 salivary protein genes, among which 9 genes (NlShpa, Salivap 3, CA, NlSEF1, Nl12, NlHSC70-3, NlSP1, NlG14, and NlDNAJB9) showed significant differences. Most differentially expressed genes are found important for BPH physiological processes, except Nl12. Here we found that silencing Nl12 impeded ovary development, thereby inhibiting oocyte formation. The potential explanation was that Nl12 was highly expressed in both salivary gland and ovary, and the ovary development abnormality may be due to the direct effect from expression reduction in ovary and/or indirect influence from expression reduction in salivary gland. Altogether, our findings provide a new insight into the mechanism of action of etofenprox on insect pests and explain part of the reason why etofenprox does not stimulate reproduction in BPH.

Investigating the role of the ROS/CncC-xenobiotics signaling pathway in the response to Fenpropathrin in Cyprinus carpio lymphocytes: Involving lipid peroxidation and Fe2+ metabolism imbalance.

Fenpropathrin (FPT) is a synthetic pyrethroid insecticide, the persistence and accumulation in water of which could cause harmful effects on vulnerable groups like aquatic creatures, particularly posing significant risks to fish immune systems. This study aimed to investigate how environmentally relevant FPT concentrations (10-1000 µ/M) affect lipid peroxidation and Fe2+ metabolism in Cyprinus carpio head kidney lymphocytes, and its relationship with oxidative stress and immunotoxicity. Firstly, CCK-8 results demonstrated that FPT caused a significant increase in lymphocyte death. Secondly, lymphocytes exposed to FPT could lead ferroptosis in lymphocytes, accompanied by evidence of the Fe2+ transporter imbalance, lipid peroxidation, Fe2+ accumulation and ferroptosis related protein increment. Thirdly, we found that FPT esposure leads to a decrease in ATP, mitochondrial DNA and NADPH/NADP+ levels, and the mRNA associated with mitochondrial function-related genes (Fis1, Drp1, and OPA1) in lymphocytes. Additionally, FPT induced the increased the levels of inflammatory genes (TNF-α, IFN-γ, and IL-6) in head kidney lymphocytes. Importantly, exposure to FPT induced oxidative stress to produce intracellular ROS, disrupting the function of the CncC signaling pathway and expression disorder of xenobiotics detoxification (CYP 450 family) genes. Notably, Treatment with NAC (a ROS inhibitor, 5 µM) demonstrated that inhibiting ROS alleviated FPT-induced lymphocyte ferroptosis and inflammatory response via the ROS/CncC-xenobiotics signaling pathway. These findings not only introduces a novel approach to investigating the immunotoxicity of FPT but also offers critical insights into mitigating the adverse effects of FPT on aquatic animal health.

Resmethrin disrupts mitochondria-associated membranes and activates endoplasmic reticulum stress, leading to proliferation inhibition in cultured mouse Leydig and Sertoli cells.

Resmethrin, a pyrethroid pesticide used to control insects, is toxic to non-target organisms and other mammals. However, little is known about the reproductive toxicity of resmethrin in the testes, or its mechanism of toxicity. In this study, we investigated the testicular toxicity of resmethrin on mouse Leydig (TM3) and Sertoli (TM4) cells, focusing on the mitochondria and endoplasmic reticulum (ER). We found that resmethrin inhibited proliferation and cell cycle progression and disrupted mitochondrial membrane potential (MMP; ΔΨ) in TM3 and TM4 cells. In particular, resmethrin exposure significantly reduced the expression of mitochondria-associated membranes (MAMs) proteins, such as Vapb, Vdac, and Grp75, in both cell lines. Resmethrin also disrupts calcium homeostasis in the mitochondrial matrix and cytoplasm. In addition, resmethrin activates oxidative stress-mediated ER stress signals. Finally, we confirmed that 4-PBA, an ER stress inhibitor, restored the growth of TM3 and TM4 cells, which was decreased by resmethrin. Therefore, we confirmed that resmethrin hampered MAMs and activated ER stress, thus suppressing TM3 and TM4 cell proliferation.