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.

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.

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.

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.

Imidacloprid-mediated alterations on the salivary glands of the Neotropical brown stink bug, Euschistus heros.

The management of the Neotropical brown stinkbug Euschistus heros (Hemiptera: Pentatomidae) in soybean fields has been heavily dependent on the application of neonicotinoid insecticides. Neonicotinoids act primarily by impairing the function of the nicotinic acetylcholine receptors of the nervous system. These compounds also target specific organs (e.g., salivary glands), which may potentiate their insecticidal efficacy. Here, we evaluated whether the exposure to the neonicotinoid imidacloprid would cause cytomorphological changes in the salivary glands of E. heros. First, we determined the lethal concentrations (LCs) of imidacloprid through contact and ingestion. Subsequently, the cytomorphology of the salivary glands were evaluated in insect groups that survived exposure to the LC5 (3.75 mg a.i./L), LC50 (112.5 mg a.i./L), or LC75 (375.0 mg a.i./L, equivalent to the recommended field rate) doses. Imidacloprid induced apoptosis and necrosis in the salivary gland cells according to the insecticide concentration and salivary gland region. All concentrations increased apoptosis and injured cells (e.g., vacuolization, chromatin condensation, swelling of organelles, and plasma membrane rupture) in the principal and accessory salivary glands. Individuals that survived exposure to the highest concentrations (i.e., LC5 and LC50) were more affected, and exhibited several necrotic cells on their main principal salivary glands. Collectively, our results indicate that imidacloprid exerts toxic effects on the non-target organs, such as the salivary glands, which increases the efficacy of this compound in the management of stink bug infestations.

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.

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars.

Soybean is one of most consumed and produced grains in the world, and Anticarsia gemmatalis is a pest that causes great damage to this crop due to severe defoliation during its larval phase. Plants have mechanisms that lead to the inhibition of proteases in the intestine of these herbivores, hampering their development. Understanding this complex protease inhibitor is important for pest control. The objective of this study was to evaluate the enzymatic profiles of the intestinal proteases of the soybean caterpillar at different instars. For this, the proteolytic profile of the gut in the third, fourth, and fifth instars were analyzed. Irreversible inhibitors of proteases were separately incubated with A. gemmatalis enzyme extracts at the third, fourth, and fifth instar to assess the contribution of these proteases to total proteolytic activity. The enzymatic extracts were also evaluated with specific substrates to confirm changes in the specific activities of trypsin-like, chymotrypsin-like, and cysteine proteases at different instars. The results showed that the protease profile of A. gemmatalis gut changes throughout its larval development. The activity of cysteine proteases was more intense in the first instar. On the contrary, the serine proteases showed major activities in the late stages of the larval phase. Zymogram analysis and protein identification by liquid chromatography-mass spectrometry indicated serine protease as the main protease class expressed in the fifth instar. These results may shift the focus from the rational development of the protease inhibitor to A. gemmatalis and other Lepidoptera, as the expression of major proteases is not constant.

"Armed to the teeth": The multiple ways to survive insecticidal and predatory challenges in Aedes aegypti larvae.

Environmental pollutants, such as insecticides, can alter the equilibrium of aquatic ecosystems, particularly those closely located to human occupations. The use of such anthropogenic compounds frequently results in the selection of resistant individuals. However, how the underlying insecticide resistance mechanisms interplay with the abilities of the resistant individuals to cope with other environmental challenges (e.g., predators) has not received adequate attention. Here, we evaluated whether resistance to pyrethroid insecticides in larvae of the yellow fever mosquito, Aedes aegypti (Diptera: Culicidae), would affect their abilities to survive other environmental challenges. We assessed the susceptibilities of the pyrethroid-resistant larvae to other insecticides (i.e., the oxadiazine indoxacarb and juvenile hormone mimic pyriproxyfen) and determined the activities of potential detoxification enzymes. Finally, we also recorded potential alterations in larva swimming behavior in the presence of predators, such as the water bug Belostoma anurum (Hemiptera: Belostomatidae). Our results revealed that high pyrethroid resistance was associated with moderate resistance to the other two insecticides. Furthermore, this multiple resistance was associated with higher detoxification activity by glutathione-S-transferases and general esterases. Interestingly, in comparison with insecticide-susceptible larvae, the pyrethroid-resistant larvae not only swam for longer periods and distances, but also took longer to be captured by B. anurum nymphs. Collectively, our findings revealed increased abilities to survive natural environmental challenges (e.g., predatory attacks) in mosquito larvae that express physiological and behavioral changes associated with multiple resistance to insecticides.

Bti-based insecticide enhances the predatory abilities of the backswimmer Buenoa tarsalis (Hemiptera: Notonectidae).

The backswimmer Buenoa tarsalis (Hemiptera: Notonectidae) is a naturally occurring predator of immature stages of mosquitoes. These aquatic predators can suffer from non-targeted exposure to insecticides that are commonly used in aquatic environments to control mosquitoes. Here, we evaluated whether insecticide formulations containing the bacterium Bacillus thuringiensis var. israelensis (Bti) or the organophosphate pirimiphos-methyl would affect the survival and the predatory abilities of B. tarsalis. First, we conducted survival bioassays to estimate the median survival time (LT50) of B. tarsalis when exposed to Bti-based insecticide (at 0.25 and 25 mg a.i./L) and pirimiphos-methyl (at 1, 10 and 1000 mg a.i./L). The highest concentrations of the insecticides were equivalent to the label-recommended field rates. Second, the predatory abilities of B. tarsalis exposed to insecticides were evaluated at three prey densities (3, 6 and 9 mosquito larvae/100 mL water) just after insecticide exposure or after a 24 h recovery time. While the survival of B. tarsalis was significantly reduced with pirimiphos-methyl concentrations ≥10 mg a.i./L, the Bti-exposed predators exhibited similar survival as unexposed predators. Interestingly, after a recovery time of 24 h, B. tarsalis sublethally exposed to pirimiphos-methyl or Bti-based insecticide consistently killed more A. aegypti larvae (at the intermediate density) than unexposed predators. However, for the without-recovery bioassays, the pirimiphos-methyl-exposed predators exhibited reduced predatory abilities at the lowest prey density. Because they do not reduce the survival or the predatory abilities of B. tarsalis, Bti-based insecticides can be considered a safe insecticide to use in the presence of backswimmers.

Toxicity of vegetable oils to the coconut mite Aceria guerreronis and selectivity against the predator Neoseiulus baraki.

The coconut mite, Aceria guerreronis (Acari: Eriophyidae), is a major tropical pest of coconut. Here, we assessed the chemical profiles and the potential use of babassu, degummed soybean, and coconut oils to control A. guerreronis as well as their side-effects on the predatory mite Neoseiulus baraki (Acari: Phytoseiidae), a key natural enemy of the coconut mite. Babassu and coconut oils had similar fatty acids chemical profiles. All vegetable oils showed toxicity to A. guerreronis; degummed soybean oil exhibited the highest toxicity (LC50 = 0.15 µL/cm2). Although all oils were less toxic to N. baraki, their potential to attract/repel this predatory mite differed. Whereas N. baraki females were unresponsive to coconut oil at both concentrations (i.e., LC50 and LC99 estimated for A. guerreronis), irrespective of exposure period (i.e., 1 or 24 h), the babassu oil repelled the predator, independent of exposure period, when applied at its LC99 (1.48 µL/cm2). Intriguingly, this oil also exhibited attractiveness to N. baraki 24 h after exposure when applied at its LC50 (0.26 µL/cm2). A similar attractiveness pattern was recorded 24 h after N. baraki was exposed to degummed soybean oil at both concentrations tested (LC50 = 0.15 µL/cm2; LC99 = 1.39 µL/cm2). However, N. baraki was repelled by degummed soybean oil at its LC50 after 1 h of exposure. Therefore, the present study demonstrated that all the vegetable oils used here had higher toxicity to the coconut mite and considerable selectivity to the predator N. baraki, indicating they are promising tools that can potentially be included in management programs to control A. guerreronis in commercial coconut plantations.

Locomotory and physiological responses induced by clove and cinnamon essential oils in the maize weevil Sitophilus zeamais.

Plant essential oils have been suggested as a suitable alternative for controlling stored pests worldwide. However, very little is known about the physiological or behavioral responses induced by these compounds in insect populations that are resistant to traditional insecticides. Thus, this investigation evaluated the toxicity (including the impacts on population growth) as well as the locomotory and respiratory responses induced by clove, Syzygium aromaticum L., and cinnamon, Cinnamomum zeylanicum L., essential oils in Brazilian populations of the maize weevil Sitophilus zeamais. We used populations that are resistant to phosphine and pyrethroids (PyPhR), only resistant to pyrethroids (PyR1 and PyR2) or susceptible to both insecticide types (SUS). The PyPhR population was more tolerant to cinnamon essential oil, and its population growth rate was less affected by both oil types. Insects from this population reduced their respiratory rates (i.e., CO2 production) after being exposed to both oil types and avoided (in free choice-experiments) or reduced their mobility on essential oil-treated surfaces. The PyR1 and PyR2 populations reduced their respiratory rates, avoided (without changing their locomotory behavior in no-choice experiments) essential oil-treated surfaces and their population growth rates were severely affected by both oil types. Individuals from SUS population increased their mobility on surfaces that were treated with both oil types and showed the highest levels of susceptibility to these oils. Our findings indicate that S. zeamais populations that are resistant to traditional insecticides might have distinct but possibly overlapping mechanisms to mitigate the actions of essential oils and traditional insecticides.

Metabolic and Behavioral Mechanisms of Indoxacarb Resistance in Sitophilus zeamais (Coleoptera: Curculionidae).

The control of the most important pest of stored maize, the weevil Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), is mainly achieved with the use of pyrethroid insecticides. However, the intensive use of these compounds has led to the selection of resistant populations and has compromised the control efficacy of this insect pest. Here, the toxicity of indoxacarb for a potential use in the control of S. zeamais was assessed on 13 Brazilian populations. Concentration-mortality bioassays, in the presence of synergists (piperonyl butoxide, triphenyl phosphate, and diethyl maleate), were used to assess potential metabolic-based indoxacarb resistance mechanisms. We also assessed the behavioral (locomotory) responses of these populations to indoxacarb exposure. The results showed significant differences between the populations (LD50 values ranged from 0.06 to 13.99 mg a.i/kg of grains), resulting in resistance ratios of >200-fold between the least (Canarana-MT) and the most (Espirito Santo do Pinhal-SP) susceptible populations. The results obtained with synergized indoxacarb suggest the involvement of esterases and glutathione-S-transferases on indoxacarb action, and also suggest the involvement of cytochrome P450-dependent monooxygenases as a potential indoxacarb resistance mechanism in Brazilian populations of S. zeamais. Although indoxacarb-induced behavioral avoidance varied among populations, some resistant populations (e.g., Canarana-MT) were able to reduce exposure to indoxacarb by spending more time in the nontreated areas. Collectively, our findings indicate that the behavioral (locomotory) and physiological responses of these insects may compromise the control efficacy of oxadiazine insecticides (e.g., indoxacarb) in Brazilian populations of S. zeamais.

Sublethal Exposure to Clove and Cinnamon Essential Oils Induces Hormetic-Like Responses and Disturbs Behavioral and Respiratory Responses in Sitophilus zeamais (Coleoptera: Curculionidae).

Essential oils have been suggested as suitable alternatives for controlling insect pests. However, the potential adaptive responses elicited in insects for mitigating the actions of these compounds have not received adequate attention. Furthermore, as is widely reported with traditional insecticides, sublethal exposure to essential oils might induce stimulatory responses or contribute to the development of resistance strategies that can compromise the management of insect pests. The current study evaluated the locomotory and respiratory responses as well as the number of larvae per grain produced by the maize weevil, Sitophilus zeamais Motschulsky, after being sublethally exposed to the essential oils of clove, Syzygium aromaticum L., and cinnamon, Cinnamomum zeylanicum L. The essential oils showed similar insecticidal toxicity (exposure route: contact with dried residues; Clove LC95 = 3.96 [2.78-6.75] µl/cm(2); Cinnamon LC95 = 3.47 [2.75-4.73] µl/cm(2)). A stimulatory effect on the median survival time (TL50) was observed when insects were exposed to low concentrations of each oil. Moreover, a higher number of larvae per grain was produced under sublethal exposure to clove essential oil. S. zeamais avoided the treated areas (in free-choice experiments) and altered their mobility when sublethally exposed to both essential oils. The respiratory rates of S. zeamais (i.e., CO2 production) were significantly reduced under low concentrations of the essential oils. We recommend the consideration of the potential sublethal effects elicited by botanical pesticides during the development of integrated pest management programs aiming to control S. zeamais.

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.

Diversity, Prevalence and Virulence of Colletotrichum Species Causing Anthracnose on Cassava Leaves in the Northern Region of Brazil.

Cassava (Manihot esculenta Crantz) is a staple crop widely cultivated by small farmers in tropical countries. However, despite the low level of technology required for its management, it can be affected by several diseases, with anthracnose as the main threat. There is little information about the main species of Colletotrichum that infect cassava in Brazil. Thus, the objective of this work was to study the diversity, prevalence and virulence of Colletotrichum species that cause anthracnose in cassava leaves in northern Brazil. Twenty municipalities of the Pará and Tocantins states were selected, and leaves with symptoms were collected in those locations. Pure cultures were isolated in the laboratory. Species were identified using phylogenetic analyses of multiple loci, and their pathogenicity, aggressivity and virulence levels were assessed. Our results showed the greatest diversity of Colletotrichum associated with anthracnose in cassava plants of the "Formosa" cultivar in the Tocantins and Pará states. We determined the presence of Colletotrichum chrysophilum, C. truncatum, C. siamense, C. fructicola, C. plurivorum, C. musicola and C. karsti, with C. chrysophilum as the most aggressive and virulent. Our findings provide accurate identifications of species of Colletotrichum causing anthracnose in cassava crops, which are of great relevance for cassava breeding programs (e.g., the search for genotypes with polygenic resistance since the pathogen is so diverse) and for developing anthracnose management strategies that can work efficiently against species complexes of Colletotrichum.

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.

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.

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.

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.