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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.

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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.

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Despite being effective in controlling mosquito larvae and a few other target organisms, the application of insecticides into aquatic systems may cause unintended alterations to the physiology or behavioral responses of several aquatic non-target organisms, which can ultimately lead to their death. Here, we firstly evaluated whether the susceptibility of the giant water bug, Belostoma anurum (Hemiptera: Belostomatidae), a predator of mosquito larvae, to pyriproxyfen would be similar to that of its potential prey, larvae of Aedes aegypti (Diptera: Culicidae). Secondly, we recorded the nominal concentrations of pyriproxyfen in water and evaluated whether sublethal exposures would lead to physiological or behavioral alterations on the B. anurum nymphs. We characterized the activities of three major families of detoxification enzymes (i.e., cytochrome P450 monooxygenases, glutathione-S-transferase, and general esterases) and further evaluated the abilities of pyriproxyfen sublethally-exposed B. anurum to prey upon A. aegypti larvae at different prey densities. Our findings revealed that nominal pyriproxyfen concentration significantly decreased (approximately 50%) over the first 24 h. Furthermore, when applied at the concentration of 10 µg a.i./L, pyriproxyfen was approximately four times more toxic to A. aegypti larvae (LT50 = 48 h) than to B. anurum nymphs (LT50 = 192 h). Interestingly, the pyriproxyfen sublethally-exposed (2.5 µg a.i./L) B. anurum nymphs exhibited reduced enzyme activities (cytochrome P450 monooxygenases) involved in detoxication processes and preyed significantly less on A. aegypti larvae when compared to unexposed predators. Collectively, our findings demonstrate that mortality-based pyriproxyfen risk assessments are not always protective of aquatic non-target organisms.

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Fishes can change their physiological responses when threatened by the presence of predators. Such physiological plasticity, however, usually implies costs that may impede organismal development and reproduction and reduce the ability to cope with other biotic and abiotic stresses. Here, we evaluated the growth and stress biomarker responses in sexually reversed Nile tilapia, Oreochromis niloticus, fingerlings indirectly threatened by the presence of the aquatic insect predator Belostoma anurum (Hemiptera: Belostomatidae). We also evaluated whether the presence of B. anurum would affect growth in fingerlings that received food containing the masculinizing hormone 17 α-methyltestosterone. The antioxidant responses were evaluated by measuring the activity of enzymes (e.g., superoxide dismutase, catalase, and glutathione-S-transferase). Oxidative stress biomarkers (e.g., malondialdehyde and nitric oxide) and blood glucose and lactate responses were also evaluated. Our results revealed that predator exposure did not affect growth in O. niloticus fingerlings reared in the presence or absence of the masculinizing hormone. However, sexually reversed tilapia fingerlings significantly increased not only the glucose and lactate blood levels, but also exhibited increased activities of superoxide dismutase and glutathione-S-transferases enzymes when threatened by the presence of B. anurum nymphs. Collectively, our findings indicate that despite not exhibiting reduced growth performance, sexually reversed tilapia fingerlings were physiologically stressed by the presence of the predator, which may reduce their ability to face environmental and abiotic stresses.

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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.

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Target-site mutations and changes in insect metabolism or behavior are common mechanisms in insecticide-resistant insects. The co-occurrence of such mechanisms in a pest strain is a prominent threat to their management, particularly when alternative compounds are scarce. Pyrethroid resistance among stored grain weevils (i.e., Sitophilus spp.) is an example of a long-standing concern, for which reports of resistance generally focus on a single mechanism in a single species. Here, we investigated pyrethroid resistance in maize and rice weevils (i.e., Sitophilus zeamais and S. oryzae), exploring potential knockdown resistance (kdr) mutations in their sodium channels (primary site for pyrethroid actions) and potential changes in their detoxification and walking processes. Resistance in pyrethroid-resistant rice weevils was associated with the combination of a kdr mutation (L1014F) and increases in walking and detoxification activities, while another kdr mutation (T929I) combined with increases in walking activity were the primary pyrethroid resistance mechanisms in maize weevils. Our results suggest that the selection of pyrethroid-resistant individuals in these weevil species may result from multiple and differential mechanisms because the L1014F mutation was only detected in Latin American rice weevils (e.g., Brazil, Argentina and Uruguay), not in Australian and Turkish rice weevils or Brazilian maize weevils.

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Freshwater ecosystems provide environmental conditions for many arthropod species, including pests like mosquitoes and beneficial insects. Giant water bugs, Belostoma anurum (Hemiptera: Belostomatidae), are aquatic insects that provide biological control of mosquitoes and small vertebrates in freshwater environments. However, the application of insecticides aiming to control mosquitoes can lead to insecticide exposures of aquatic predators that can result in their death or significant reductions in their behavioral abilities. Here, we assessed the susceptibilities of B. anurum to the pyrethroid insecticide deltamethrin and evaluated whether sublethal exposure to deltamethrin would change the abilities of B. anurum to prey upon larvae of Aedes aegypti (Diptera: Culicidae). Bioassays of predator performance were conducted at three prey densities (i.e., 3, 6 and 9 larvae/100 mL of water) just after insecticide exposure and on the three following days. Our results revealed that B. anurum (LC50 = 90.9 µg a. i./L) was approximately 32-fold less susceptible to deltamethrin than A. aegypti larvae (LC50 = 2.8 µg a. i./L). However, the number of larvae eaten by B. anurum sublethally exposed to deltamethrin (at 13 µg a. i./L for 24 h) was significantly (P < 0.05) smaller than that recorded for unexposed predators. Furthermore, the deltamethrin-mediated behavioral changes were higher at the highest availability of prey and, as expected, just after insecticide exposure. Thus, sublethal exposure to deltamethrin reduces the ability of B. anurum to capture and prey upon A. aegypti larvae, compromising the efficacy of these insects as naturally occurring mosquito control agents.

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The yellow fever mosquito, Aedes aegypti, particularly in Neotropical regions, is the principal vector of dengue, yellow fever, Zika and Chikungunya viruses. Pyrethroids remain one of the most used insecticides to control Aedes mosquitoes, despite the development of pyrethroid resistance in many mosquito populations worldwide. Here, we report a Brazilian strain of A. aegypti with high levels (approximately 100-60,000 fold) of resistance to both type I and type II pyrethroids. We detected two mutations (V410L and F1534C) in the sodium channel from this resistant strain. This study is the first report of the V410L mutation in mosquitoes. Alone or in combination with the F1534C mutation, the V410L mutation drastically reduced the sensitivity of mosquito sodium channels expressed in Xenopus oocytes to both type I and type II pyrethroids. The V410L mutation presents a serious challenge for the control of A. aegypti and will compromise the use of pyrethroids for the control of A. aegypti in Brazil; therefore, early monitoring of the frequency of the V410L mutation will be a key resistance management strategy to preserve the effectiveness of pyrethroid insecticides.

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The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), one of the principal pests of brassicas, can reduce productivity and thus cause losses for brassica farmers. Alternative controls, such as biological agents and plant extracts, may be used to reduce insect populations, either alone or in conjunction with pest management programs. The objective was to evaluate the insecticidal activity of Beauveria bassiana (Bals.) Vuill. and castor bean oil and mixtures of both components against the diamondback moth. To do so, we separately used castor bean oil (at 2% concentration), the isolate ESALQ-447 and a commercial formulation (Boveril® WP), and a mixture of castor bean oil with the isolate and the B. bassiana product formulation, totaling six treatments with a control. Assays were carried out under greenhouse with the respective treatments sprayed on cabbage plants infested with four second instar larvae of P. xylostella. The evaluated parameters were larval mortality and pupal and larval viability. All treatments reduced larval viability in relation to the control, however, only the ESALQ-447 isolate or a mixture of the isolate with castor bean oil reduced pupal viability, significantly reducing the pest population levels in the next generation. Castor bean oil mixed with B. bassiana, however, does not augment pest mortality.

A traça-das-crucíferas, Plutella xylostella (L.) (Lepidoptera: Plutellidae), uma das principais pragas das brássicas, pode ocasionar redução na produtividade e consequentemente prejuízos aos produtores rurais destas culturas. A busca por alternativas de controle, como o uso do controle biológico e extratos de plantas são métodos de redução populacional de insetos que podem ser usados isoladamente ou associados em programas de manejo fitossanitário. Dessa forma, o objetivo foi avaliar a atividade inseticida de Beauveria bassiana (Bals.) Vuill., do óleo de mamona e a associação destes, visando ao controle da traça-das-crucíferas. Para isso, foram utilizados isoladamente o óleo de mamona (na concentração de 2%), o isolado ESALQ-447 e um formulado comercial (Boveril® WP) e a mistura do óleo de mamona com o respectivo isolado e com o produto formulado de B. bassiana, totalizando assim seis tratamentos com a testemunha. Os ensaios foram realizados em casa telada, onde plantas de repolho infestadas com quatro larvas de segundo ínstar de P. xylostella foram pulverizadas com os respectivos tratamentos mencionados. Os parâmetros avaliados foram mortalidade larval, viabilidade larval e pupal. Todos os tratamentos reduziram a viabilidade larval em relação à testemunha, no entanto apenas o isolado ESALQ-447 ou a sua associação com o óleo de mamona reduziram a viabilidade pupal, diminuindo significativamente o nível populacional da praga na próxima geração. Contudo, o óleo de mamona misturado com B. bassiana não aumenta a mortalidade da praga.

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