Potential of Ocotea indecora Essential Oil for Controlling Drosophila suzukii: Molecular Predictions for Toxicity and Selectivity to Beneficial Arthropods.

The protection of soft-skinned fruits against Drosophila suzukii has relied primarily on the efficacy of a few synthetic molecules. Despite their short-term efficacy, these molecules can cause environmental pollution, unintendedly affect non-target organisms, and fail to provide sustainable control. The shortfalls of using synthetic pesticides warrant the search for alternatives, such as essential oils extracted from plants, with greater eco-friendlier properties. Here, we chemically characterized and evaluated the toxicity of the essential oil extracted from leaves of Ocotea indecora (Schott) Mez (Lauraceae) against D. suzukii via two exposure pathways (ingestion and contact). We also assessed the selectivity of the essential oil to two predatory natural enemies, Eriopis connexa and Chrysoperla externa and two pollinator bees, Apis mellifera and Partamona helleri. In addition, we conducted in silico predictions to investigate potential interactions between the major compound of the essential oil and the insects' transient receptor potential (TRP) channels. Our chromatographic analysis revealed sesquirosefuran (87%) as the major compound. Higher toxicity to adults of D. suzukii was observed in contact exposure (LC50 = 0.43 µL mL-1) compared to ingestion (LC50 = 0.72 µL mL-1). However, the essential oil did not cause mortality to the non-target organisms tested here, even when applied at 2.20 µL mL-1. Molecular predictions demonstrated that sesquirosefuran binds more stably to the TRP channels of D. suzukii than to those expressed in beneficial arthropods. Collectively, our findings provide the initial framework for the potential use of O. indecora essential oil as a sustainable alternative for managing D. suzukii infestations.

Draft genome of neotropical Bacillus thuringiensis UFT038 and its potential against lepidopteran soybean pests.

Bacillus thuringiensis (Bt) is known for its Cry and Vip3A pesticidal proteins with high selectivity to target pests. Here, we assessed the potential of a novel neotropical Bt strain (UFT038) against six lepidopteran pests, including two Cry-resistant populations of fall armyworm, Spodoptera frugiperda. We also sequenced and analyzed the genome of Bt UFT038 to identify genes involved in insecticidal activities or encoding other virulence factors. In toxicological bioassays, Bt UFT038 killed and inhibited the neonate growth in a concentration-dependent manner. Bt UFT038 and HD-1 were equally toxic against S. cosmioides, S. frugiperda (S_Bt and R_Cry1 + 2Ab populations), Helicoverpa zea, and H. armigera. However, larval growth inhibition results indicated that Bt UFT038 was more toxic than HD-1 to S. cosmioides, while HD-1 was more active against Chrysodeixis includens. The draft genome of Bt UFT038 showed the cry1Aa8, cry1Ac11, cry1Ia44, cry2Aa9, cry2Ab35, and vip3Af5 genes. Besides this, genes encoding the virulence factors (inhA, plcA, piplC, sph, and chi1-2) and toxins (alo, cytK, hlyIII, hblA-D, and nheA-C) were also identified. Collectively, our findings reveal the potential of the Bt UFT038 strain as a source of insecticidal genes against lepidopteran pests, including S. cosmioides and S. frugiperda.

Novel Lactone-Based Insecticides and Drosophila suzukii Management: Synthesis, Potential Action Mechanisms and Selectivity for Non-Target Parasitoids.

Drosophila suzukii, an invasive insect pest, poses a significant threat to various fruit crops. The use of broad-spectrum insecticides to control this pest can reduce the effectiveness of biological control agents, such as the parasitoid Trichopria anastrephae. Here, we evaluated the toxicity of newly synthesized lactone derivatives on D. suzukii and their selectivity towards T. anastrephae. We used in silico approaches to identify potential targets from the most promising molecules in the D. suzukii nervous system and to understand potential differences in susceptibilities between D. suzukii and its parasitoid. Of the nine molecules tested, (rac)-8 and compound 4 demonstrated efficacy against the fly. Exposure to the estimated LC90 of (rac)-8 and compound 4 resulted in a mortality rate of less than 20% for T. anastrephae without impairing the parasitoid's functional parasitism. The in silico predictions suggest that (rac)-8 and compound 4 target gamma amino butyric acid (GABA) receptors and transient receptor potential (TRP) channels of D. suzukii. However, only the reduced interaction with TRP channels in T. anastrephae demonstrated a potential reason for the selectivity of these compounds on the parasitoid. Our findings suggest the potential for integrating (rac)-8 and compound 4 into D. suzukii management practices.

Acute sublethal exposure to ethiprole impairs physiological and oxidative status in the Neotropical fish Astyanax altiparanae.

Ethiprole, a phenylpyrazole insecticide, has been increasingly used in the Neotropical region to control stink bug pests in soybean and maize fields. However, such abrupt increases in use may have unintended effects on non-target organisms, including those inhabiting freshwater ecosystems. Here, we evaluated the effects of acute (96 h) sublethal exposure to ethiprole (up to 180 µg/L, which is equivalent to 0.013% of the recommended field dose) on biomarkers of stress in the gills, liver, and muscle of the Neotropical fish Astyanax altiparanae. We further recorded potential ethiprole-induced effects on the structural histology of A. altiparanae gills and liver. Our results showed that ethiprole exposure increased glucose and cortisol levels in a concentration-dependent manner. Ethiprole-exposed fish also exhibited higher levels of malondialdehyde and greater activity of antioxidant enzymes, such as glutathione-S-transferase and catalase, in both gills and liver. Furthermore, ethiprole exposure led to increased catalase activity and carbonylated protein levels in muscle. Morphometric and pathological analyses of the gills revealed that increasing ethiprole concentration resulted in hyperemia and loss of integrity of the secondary lamellae. Similarly, histopathological analysis of the liver demonstrated higher prevalence of necrosis and inflammatory infiltrates with increasing ethiprole concentration. Altogether, our findings demonstrated that sublethal exposure to ethiprole can trigger a stress response in non-target fish species, which may lead to potential ecological and economic imbalances in Neotropical freshwater systems.

Sex-specific alterations in adaptive responses of Chironomus columbiensis triggered by imidacloprid chronic and acute sublethal exposures.

The use of imidacloprid is a common pest control practice in the Neotropical region. However, the imidacloprid unintended sublethal effects on Neotropical aquatic non-target arthropods and undesirable consequences for aquatic environments remain unclear. Here, we assessed the susceptibility of Chironomus columbiensis (Diptera: Chironomidae) larvae to the neonicotinoid imidacloprid and evaluated whether sublethal exposure types would trigger sex-dependent adaptive responses (e.g., emergence, body mass, reproduction, wing morphology). We conducted a concentration-mortality curve (96 h of exposure) and established chronic and acute sublethal exposure bioassays. While chronic sublethal exposures consisted of exposing individuals during their entire larval and pupal stages, the acute sublethal exposures represented a single short duration (24 h) exposure episode during either the first or fourth larval instar. Our results revealed that chronic sublethal exposure reduced the body mass of males, while acute sublethal exposures during the first instar resulted in heavier males than those that were not exposed to imidacloprid. Chronic exposure also reduced the reproduction of males and females, while the acute sublethal exposure only affected the reproduction of individuals that were imidacloprid-exposed on their later larval instar. Chronic and acute sublethal exposures did differentially affect the wing properties of C. columbiensis males (e.g., increased size when chronically exposed and highly asymmetric wings when acutely exposed in early larval phase) and females (e.g., highly asymmetric wings when chronically and acutely exposed). Collectively, our findings demonstrated that imidacloprid can cause unintended sublethal effects on C. columbiensis, and those effects are dependent on sex, exposure type, and developmental stage.

Using an Aluminum Hydroxide-Chitosan Matrix Increased the Vaccine Potential and Immune Response of Mice against Multi-Drug-Resistant Acinetobacter baumannii.

Acinetobacter baumannii is a Gram-negative, immobile, aerobic nosocomial opportunistic coccobacillus that causes pneumonia, septicemia, and urinary tract infections in immunosuppressed patients. There are no commercially available alternative antimicrobials, and multi-drug resistance is an urgent concern that requires emergency measures and new therapeutic strategies. This study evaluated a multi-drug-resistant A. baumannii whole-cell vaccine, inactivated and adsorbed on an aluminum hydroxide-chitosan (mAhC) matrix, in an A. baumannii sepsis model in immunosuppressed mice by cyclophosphamide (CY). CY-treated mice were divided into immunized, non-immunized, and adjuvant-inoculated groups. Three vaccine doses were given at 0D, 14D, and 28D, followed by a lethal dose of 4.0 × 108 CFU/mL of A. baumannii. Immunized CY-treated mice underwent a significant humoral response, with the highest IgG levels and a higher survival rate (85%); this differed from the non-immunized CY-treated mice, none of whom survived (p < 0.001), and from the adjuvant group, with 45% survival (p < 0.05). Histological data revealed the evident expansion of white spleen pulp from immunized CY-treated mice, whereas, in non-immunized and adjuvanted CY-treated mice, there was more significant organ tissue damage. Our results confirmed the proof-of-concept of the immune response and vaccine protection in a sepsis model in CY-treated mice, contributing to the advancement of new alternatives for protection against A. baumannii infections.

Methanolic Extracts of Chiococca alba in Aedes aegypti Biorational Management: Larvicidal and Repellent Potential, and Selectivity against Non-Target Organisms.

The use of formulations containing botanical products for controlling insects that vector human and animal diseases has increased in recent years. Plant extracts seem to offer fewer risks to the environment and to human health without reducing the application strategy's efficacy when compared to synthetic and conventional insecticides and repellents. Here, we evaluated the potential of extracts obtained from caninana, Chiococca alba (L.) Hitchc. (Rubiaceae), plants as a tool to be integrated into the management of Aedes aegypti, one of the principal vectors for the transmission of arborviruses in humans. We assessed the larvicidal and repellence performance against adult mosquitoes and evaluated the potential undesired effects of the extracts on non-target organisms. We assessed the susceptibility and predatory abilities of the nymphs of Belostoma anurum, a naturally occurring mosquito larva predator, and evaluated the C. alba extract's cytotoxic effects in mammalian cell lines. Our chromatographic analysis revealed 18 compounds, including rutin, naringin, myricetin, morin, and quercetin. The methanolic extracts of C. alba showed larvicidal (LC50 = 82 (72-94) mg/mL) activity without killing or affecting the abilities of B. anurum to prey upon mosquito larvae. Our in silico predictions revealed the molecular interactions between rutin and the AeagOBP1 receptor to be one possible mechanism for the repellent potential recorded for formulations containing C. alba extracts. Low cytotoxicity against mammalian cell lines reinforces the selectivity of C. alba extracts. Collectively, our findings highlight the potential of C. alba and one of its constituents (rutin) as alternative tools to be integrated into the management of A. aegypti mosquitoes.

Potential Action Mechanism and Inhibition Efficacy of Morinda citrifolia Essential Oil and Octanoic Acid against Stagonosporopsis cucurbitacearum Infestations.

The use of plant-based products has been shown to efficiently inhibit fungi-mediated diseases in agricultural crops. Here, we extracted and evaluated the composition of noni, Morinda citrifolia L., essential oil and assessed its activities against Stagonosporopsis cucurbitacearum in Cucumis melo L. Using in silico molecular approaches, potential interactions between the essential oil major components and S. cucurbitacearum tyrosine-tRNA ligase were predicted. Finally, we also measured the potential interference of plant physiology (the stomatal conductance and net photosynthesis) mediated by the application of the M. citrifolia essential oil. Chromatographic analysis revealed that octanoic acid (75.8%), hexanoic acid (12.8%), and isobutyl pent-4-enyl carbonate (3.1%) were the major essential oil compounds. Octanoic acid and noni essential oil, when used as preventive measures, reduce fungal mycelial growth at a concentration of 5 mg/mL without causing significant damage to the treated leaves, which reinforces their efficacies as preventive tools against S. cucurbitacearum. Molecular docking analyses predicted very stable interactions between the major essential oil constituents and S. cucurbitacearum tyrosine-tRNA ligase, suggesting the interference of these plant-based molecules upon enzyme activation. Octanoic acid and M. citrifolia essential oil at concentrations of 20 mg/mL decreased the stomatal conductance and net photosynthesis rate of melon plants, resulting in robust phytotoxicity. Collectively, our findings indicated that despite the phytotoxicity risks at higher concentrations, M. citrifolia essential oil and octanoic acid, have potential as alternative tools for the integrative management of S. cucurbitacearum.

Sublethal exposure to pyriproxyfen does not impair the abilities of the backswimmer Buenoa amnigenus to prey upon Aedes aegypti larvae.

Pyriproxyfen is a juvenile hormone analogue that is commonly used to control the immature stages of mosquitoes in both artificial and natural water reservoirs. Recently, concerns have been raised regarding the community effectiveness of pyriproxyfen in preventing vector-transmitted diseases. Such concerns have been based on the unintended effects on non-target organisms and the selection of resistant mosquito populations. This investigation was, therefore, conducted to evaluate the toxicity of pyriproxyfen to Aedes aegypti (Diptera: Culicidae) larvae and the backswimmer Buenoa amnigenus (Hemiptera: Notonectidae), a naturally occurring mosquito larvae predator. We also assessed the abilities of backswimmers exposed to sublethal levels of pyriproxyfen to prey upon mosquito larvae (L2) under three larval densities (3, 6, or 9 larvae/100 mL of water) using artificial containers. Our results revealed that pyriproxyfen killed backswimmers only at concentrations higher than 100 µg active ingredient [a.i.]/L, which is 10 times higher than that recommended for larvicidal field application (i.e, 10 µg a.i./L). The abilities of backswimmers exposed to sublethal levels of pyriproxyfen (100 µg a.i./L) to prey upon mosquito larvae were not affected. Harmful effects on the backswimmer predatory abilities were detected only at concentrations of 150 µg a.i./L and when there was a higher prey availability (i.e., 9 larvae/100 mL of water). Together, our findings indicate that the reduced community effectiveness of this insecticide derives from factors other than its detrimental effects on non-target organisms such as backswimmers.

Risk assessment of RNAi-based pesticides to non-target organisms: Evaluating the effects of sequence similarity in the parasitoid wasp Telenomus podisi.

RNA interference (RNAi)-based pesticides are promising novel pest management products that might reduce environmental impacts compared to other pesticides. Their sequence-guided mode of action facilitates a high species-selectivity, preventing harm on non-target organisms. However, there is currently no consensus on the minimum needed sequence similarity for efficient RNAi in insects and studies have shown that adverse effects in non-targets cannot always be ruled out a priori. This study investigates the effects of exposing the parasitoid wasp Telenomus podisi to double-stranded RNA (dsRNA) which is lethal to its host, the Neotropical brown stink bug Euschistus heros. Feeding T. podisi with wasp-specific dsRNA targeting the vATPase A and actin-2 genes led to 76.4 ± 9.9% and 76.7 ± 8.8% mortality respectively, demonstrating that dietary RNAi is functional in T. podisi. When feeding T. podisi with E. heros-specific dsRNA targeting the same genes, no lethal or sublethal effects were observed. To link sequence similarity to potential gene silencing effects in the parasitoids, the expression of genes showing the highest degree of similarity (17-21 nucleotide matches) with these two target genes was monitored and was found unaffected by the E. heros-specific dsRNA. Our study confirms that RNAi was in this case highly specific and that for E. heros, RNAi-based pesticides can be used complementary to biological control in an integrated pest management context.

Bayesian Multi-Targets Strategy to Track Apis mellifera Movements at Colony Level.

Interactive movements of bees facilitate the division and organization of collective tasks, notably when they need to face internal or external environmental challenges. Here, we present a Bayesian and computational approach to track the movement of several honey bee, Apis mellifera, workers at colony level. We applied algorithms that combined tracking and Kernel Density Estimation (KDE), allowing measurements of entropy and Probability Distribution Function (PDF) of the motion of tracked organisms. We placed approximately 200 recently emerged and labeled bees inside an experimental colony, which consists of a mated queen, approximately 1000 bees, and a naturally occurring beehive background. Before release, labeled bees were fed for one hour with uncontaminated diets or diets containing a commercial mixture of synthetic fungicides (thiophanate-methyl and chlorothalonil). The colonies were filmed (12 min) at the 1st hour, 5th and 10th days after the bees' release. Our results revealed that the algorithm tracked the labeled bees with great accuracy. Pesticide-contaminated colonies showed anticipated collective activities in peripheral hive areas, far from the brood area, and exhibited reduced swarm entropy and energy values when compared to uncontaminated colonies. Collectively, our approach opens novel possibilities to quantify and predict potential alterations mediated by pollutants (e.g., pesticides) at the bee colony-level.

Exposures to deltamethrin on immature Chironomus columbiensis drive sublethal and transgenerational effects on their reproduction and wing morphology.

Sublethal exposure to insecticides can trigger unintended responses in non-target insects that may disrupt reproductive and developmental performances of these organisms. Here, we assessed whether sublethal exposure to the pyrethroid insecticide deltamethrin in early life had sublethal and transgenerational effects on the reproduction (i.e., fecundity and fertility) and wing morphology of Chironomus columbiensis, an aquatic insect used as a water quality indicator. We first conducted concentration-response bioassays to evaluate the susceptibility of C. columbiensis larvae to deltamethrin. Our results revealed that deltamethrin toxicity was approximately 7-fold higher when C. columbiensis larvae where exposed to 96 h (LC50 = 0.17 [0.15-0.20] µg/L) than to 24 h (LC50 = 1.17 [0.97-1.43] µg/L). Furthermore, the sublethal exposures (at LC1 = 0.02 µg/L or LC10 = 0.05 µg/L) of immature C. columbiensis resulted in lower fecundity (e.g., reduced eggs production) and morphometric variation wing shapes. Further reduction in fertility rates (quantity of viable eggs) occurred at deltamethrin LC10 (0.05 µg/L). Almost 80% of the fecundity was recovered with only a single recovery generation; however, two subsequent recovery generations were not sufficient to fully recover fecundity in C. columbiensis. Specimens recovered from 98.5% of wing morphometric variation after two consecutive generations without deltamethrin exposure. Collectively, our findings demonstrates that sublethal exposure to synthetic pyrethroids such as deltamethrin detrimentally affect the reproduction and wing shape of C. columbiensis, but also indicate that proper management of these compounds (e.g., concentration and frequency of application) would suffice for these insects' population recovery.

Bioallethrin activates specific olfactory sensory neurons and elicits spatial repellency in Aedes aegypti.

BACKGROUND: Use of pyrethroid insecticides is a pivotal strategy for mosquito control globally. Commonly known for their insecticidal activity by acting on voltage-gated sodium channels, pyrethroids, such as bioallethrin and transfluthrin, are used in mosquito coils, emanators and other vaporizers to repel mosquitoes and other biting arthropods. However, whether specific olfactory receptor neurons are activated by pyrethroids to trigger spatial repellency remains unknown. RESULTS: We used behavioral and electrophysiological approaches to elucidate the mechanism of bioallethrin repellency in Aedes aegypti, a major vector of dengue, yellow fever, Zika and chikungunya viruses. We found that bioallethrin elicits spatial (i.e. non-contact) repellency and activates a specific type of olfactory receptor neuron in mosquito antennae. Furthermore, bioallethrin repellency is significantly reduced in a mosquito mutant of Orco, an obligate olfactory co-receptor that is essential for the function of odorant receptors (Ors). These results indicate that activation of specific Or(s) by bioallethrin contributes to bioallethrin repellency. In addition, bioallethrin repellency was reduced in a pyrethroid-resistant strain that carries two mutations in the sodium channel gene that are responsible for knockdown resistance (kdr) to pyrethroids, indicating a role of activation of sodium channels in bioallethrin repellency. CONCLUSION: Results from this study show that bioallethrin repellency is likely to be the result of co-activation of Or(s) and sodium channels. These findings not only contribute to our understanding of the modes of action of volatile pyrethroids in spatial repellency, but also provide a framework for developing new repellents based on the dual-target mechanism revealed. © 2021 Society of Chemical Industry.

Implications of Sublethal Insecticide Exposure and the Development of Resistance on Mosquito Physiology, Behavior, and Pathogen Transmission.

For many decades, insecticides have been used to control mosquito populations in their larval and adult stages. Although changes in the population genetics, physiology, and behavior of mosquitoes exposed to lethal and sublethal doses of insecticides are expected, the relationships between these changes and their abilities to transmit pathogens remain unclear. Thus, we conducted a comprehensive review on the sublethal effects of insecticides and their contributions to insecticide resistance in mosquitoes, with the main focus on pyrethroids. We discuss the direct and acute effects of sublethal concentrations on individuals and populations, the changes in population genetics caused by the selection for resistance after insecticide exposure, and the major mechanisms underlying such resistance. Sublethal exposures negatively impact the individual's performance by affecting their physiology and behavior and leaving them at a disadvantage when compared to unexposed organisms. How these sublethal effects could change mosquito population sizes and diversity so that pathogen transmission risks can be affected is less clear. Furthermore, despite the beneficial and acute aspects of lethality, exposure to higher insecticide concentrations clearly impacts the population genetics by selecting resistant individuals, which may bring further and complex interactions for mosquitoes, vertebrate hosts, and pathogens. Finally, we raise several hypotheses concerning how the here revised impacts of insecticides on mosquitoes could interplay with vector-mediated pathogens' transmission.

Differential defense responses of tropical grasses to Mahanarva spectabilis (Hemiptera: Cercopidae) infestation.

The spittlebugs Mahanarva spectabilis economically challenges cattle production of neotropical regions, due to its voracious feeding on tropical grasses. Here, we evaluated biochemical responses of the interaction between M. spectabilis and the widely cultivated tropical grasses Brachiaria spp. (i.e., brizantha and decumbens) and elephant grasses (cvs. Roxo de Botucatu and Pioneiro), regarding lipoxygenases, protease inhibitors, phytohormones, and proteolytic activities in the midgut of M. spectabilis. The M. spectabilis-infested grasses increased lipoxygenases activity, except for cv. Pioneiro. The levels of the phytohormones jasmonic and abscisic acids were similarly low in all genotypes and increased under herbivory. Furthermore, salicylic acid concentration was constitutively higher in Brachiaria sp., increasing only in spittlebug-infested B. decumbens. M. spectabilis infestations did not induce increases of protease inhibitors in any forage grass type. The trypsin activity remained unaltered, and the total proteolytic activity increased only in B. decumbens-fed insects. Our findings revealed that most forage grasses exposed to spittlebugs activate the lipoxygenases pathway, resulting in increased abscisic and jasmonic acids. However, greater amounts of these hormones do not induce protease inhibitory activity in response to spittlebug attack. This knowledge certainly helps to guide future projects aiming at reducing the impact of spittlebugs on forage production.

Identification of multiple odorant receptors essential for pyrethrum repellency in Drosophila melanogaster.

Pyrethrum extract from dry flowers of Tanacetum cinerariifolium (formally Chrysanthemum cinerariifolium) has been used globally as a popular insect repellent against arthropod pests for thousands of years. However, the mechanistic basis of pyrethrum repellency remains unknown. In this study, we found that pyrethrum spatially repels and activates olfactory responses in Drosophila melanogaster, a genetically tractable model insect, and the closely-related D. suzukii which is a serious invasive fruit crop pest. The discovery of spatial pyrethrum repellency and olfactory response to pyrethrum in D. melanogaster facilitated our identification of four odorant receptors, Or7a, Or42b, Or59b and Or98a that are responsive to pyrethrum. Further analysis showed that the first three Ors are activated by pyrethrins, the major insecticidal components in pyrethrum, whereas Or98a is activated by (E)-ß-farnesene (EBF), a sesquiterpene and a minor component in pyrethrum. Importantly, knockout of Or7a, Or59b or Or98a individually abolished fly avoidance to pyrethrum, while knockout of Or42b had no effect, demonstrating that simultaneous activation of Or7a, Or59b and Or98a is required for pyrethrum repellency in D. melanogaster. Our study provides insights into the molecular basis of repellency of one of the most ancient and globally used insect repellents. Identification of pyrethrum-responsive Ors opens the door to develop new synthetic insect repellent mixtures that are highly effective and broad-spectrum.

Sodium channel activation underlies transfluthrin repellency in Aedes aegypti.

BACKGROUND: Volatile pyrethroid insecticides, such as transfluthrin, have received increasing attention for their potent repellent activities in recent years for controlling human disease vectors. It has been long understood that pyrethroids kill insects by promoting activation and inhibiting inactivation of voltage-gated sodium channels. However, the mechanism of pyrethroid repellency remains poorly understood and controversial. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that transfluthrin repels Aedes aegypti in a hand-in-cage assay at nonlethal concentrations as low as 1 ppm. Contrary to a previous report, transfluthrin does not elicit any electroantennogram (EAG) responses, indicating that it does not activate olfactory receptor neurons (ORNs). The 1S-cis isomer of transfluthrin, which does not activate sodium channels, does not elicit repellency. Mutations in the sodium channel gene that reduce the potency of transfluthrin on sodium channels decrease transfluthrin repellency but do not affect repellency by DEET. Furthermore, transfluthrin enhances DEET repellency. CONCLUSIONS/SIGNIFICANCE: These results provide a surprising example that sodium channel activation alone is sufficient to potently repel mosquitoes. Our findings of sodium channel activation as the principal mechanism of transfluthrin repellency and potentiation of DEET repellency have broad implications in future development of a new generation of dual-target repellent formulations to more effectively repel a variety of human disease vectors.

Potential of Ficus carica extracts against Euschistus heros: Toxicity of major active compounds and selectivity against beneficial insects.

BACKGROUND: Using plant-based extracts and their constituents has been suggested as an alternative tool to replace or integrate with the synthetic compounds used to manage insect pests. Here, we evaluated the potential of extracts obtained from Ficus carica Linn (Moraceae) branches and leaves against the Neotropical brown stink bug, Euschistus heros, one of the most prevalent insect pests in soybean fields. We further isolated and evaluated the toxicity of the extracts' major components against E. heros. Additionally, by using computational docking analysis and toxicological approaches, we assessed the physiological basis for the selectivity of these extracts against beneficial insects such as pollinator bees (i.e. Apis mellifera and the Neotropical stingless bee Partamona helleri), ladybeetles (Eriopis connexa and Coleomegilla maculata), and lacewings (Chrysoperla externa). RESULTS: Our results demonstrate that branch (LC50  = 5.9 [4.7-7.1] mg mL-1 ) and leaf (LC50  = 14.1 [12.5-15.4] mg mL-1 ) extracts exhibited similar toxicity against E. heros. Our phytochemical analysis revealed psoralen and bergapten furanocoumarins as the major components of the extract. Based on our computational predictions, these molecules' differential abilities to physically interact with the acetylcholinesterases of E. heros and beneficial insects play relevant roles in their selectivity actions. The estimated LC90 values of branch (30.0 mg mL-1 ) and leaf (30.0 mg mL-1 ) extracts killed less than 12% of the beneficial insects. CONCLUSION: Overall, our findings revealed that furanocoumarin-rich extracts obtained from F. carica extracts have the potential to be used as alternative tools in the integrated management of stink bug pests. © 2021 Society of Chemical Industry.

A dual-target molecular mechanism of pyrethrum repellency against mosquitoes.

Pyrethrum extracts from flower heads of Chrysanthemum spp. have been used worldwide in insecticides and repellents. While the molecular mechanisms of its insecticidal action are known, the molecular basis of pyrethrum repellency remains a mystery. In this study, we find that the principal components of pyrethrum, pyrethrins, and a minor component, (E)-ß-farnesene (EBF), each activate a specific type of olfactory receptor neurons in Aedes aegypti mosquitoes. We identify Ae. aegypti odorant receptor 31 (AaOr31) as a cognate Or for EBF and find that Or31-mediated repellency is significantly synergized by pyrethrin-induced activation of voltage-gated sodium channels. Thus, pyrethrum exerts spatial repellency through a novel, dual-target mechanism. Elucidation of this two-target mechanism may have potential implications in the design and development of a new generation of synthetic repellents against major mosquito vectors of infectious diseases.

Extremely Low-Frequency Electromagnetic Fields Entrain Locust Wingbeats.

Extremely low-frequency electromagnetic fields (ELF EMFs) have been shown to impact the behavior and physiology of insects. Recent studies have highlighted the need for more research to determine more specifically how they affect flying insects. Here, we ask how locust flight is affected by acute exposure to 50 Hz EMFs. We analyzed the flights of individual locusts tethered between a pair of copper wire coils generating EMFs of various frequency using high-speed video recording. The mean wingbeat frequency of tethered locusts was 18.92 ± 0.27 Hz. We found that acute exposure to 50 Hz EMFs significantly increased absolute change in wingbeat frequency in a field strength-dependent manner, with greater field strengths causing greater changes in wingbeat frequency. The effect of EMFs on wingbeat frequency depended on the initial wingbeat frequency of a locust, with locusts flying at a frequency lower than 20 Hz increasing their wingbeat frequency, while locusts flying with a wingbeat frequency higher than 20 Hz decreasing their wingbeat frequency. During the application of 50 Hz EMF, the wingbeat frequency was entrained to a 2:5 ratio (two wingbeat cycles to five EMF cycles) of the applied EMF. We then applied a range of ELF EMFs that were close to normal wingbeat frequency and found that locusts entrained to the exact frequency of the applied EMF. These results show that exposure to ELF EMFs lead to small but significant changes in wingbeat frequency in locusts. We discuss the biological implications of the coordination of insect flight in response to electromagnetic stimuli. © 2021 Bioelectromagnetics Society.