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Bees are often exposed to pesticides affecting physiological functions and molecular mechanisms. Studies showed a potential link between altered expression of energy metabolism related transcripts and increased homing flight time of foragers exposed to pesticides. In this study, we investigated the effects of thiamethoxam and pyraclostrobin on longevity, flight behavior, and expression of transcripts involved in endocrine regulation (hbg-3, buffy, vitellogenin) and energy metabolism (cox5a, cox5b, cox17) using radio frequency identification (RFID) technology and quantitative polymerase chain reaction. Parallel, a laboratory study was conducted investigating whether pesticide exposure alone without the influence of flight activity caused similar expression patterns as in the RFID experiment. No significant effect on survival, homing flight duration, or return rate of exposed bees was detected. The overall time foragers spent outside the hive was significantly reduced post-exposure. Irrespective of the treatment group, a correlation was observed between cox5a, cox5b, cox17 and hbg-3 expression and prolonged homing flight duration. Our results suggest that flight behavior can impact gene expression and exposure to pesticides adversely affects the expression of genes that are important for maintaining optimal flight capacity. Our laboratory-based experiment showed significantly altered expression levels of cox5a, cox6c, and cox17. However, further work is needed to identify transcriptional profiles responsible for prolonged homing flight duration., (© 2024. The Author(s).)

Background: Controlling the spread of arboviral diseases remains a considerable challenge due to the rapid development of insecticide resistance in Aedes mosquitoes. This study evaluated the effects of boric acid-containing toxic sugar bait (TSB) on field populations of resistant Aedes aegypti mosquitoes. In addition, this study examined the flight activity and wing beat frequency and amplitude of males and the flight activity, fecundity, and insemination of females after pairing with males exposed to TSB. The population dynamics of Aedes mosquitoes under imbalanced sex ratios were examined to simulate realistic field conditions for male suppression under the effect of TSB., Results: The mortality of male mosquitoes was consistently high within 24 h after exposure. By contrast, the mortality of female mosquitoes was inconsistent, with over 70% mortality observed at 168 h. The flight activity and wing beat amplitude of treated males were significantly lower than those of controls, but no significant difference in wing beat frequency was detected. The fecundity and insemination of treated female mosquitoes were lower than those of controls. A simulation study indicated that considerably low male population densities led to mating failures, triggering a mate-finding Allee effect and resulting in persistently low population levels., Conclusion: Boric acid-containing TSB could effectively complement current chemical intervention approaches to control resistant mosquito populations. TSB is effective in reducing field male populations and impairing male flight activity and female-seeking behavior, resulting in decreased fecundity and insemination. Male suppression due to TSB potentially results in a small mosquito population. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Imidacloprid, chlorpyrifos, and glyphosate rank among the most extensively employed pesticides worldwide. The effects of these pesticides and their combined on the flight capability of Apis cerana, and the potential underlying mechanisms remain uncertain. To investigate these effects, we carried out flight mill, transcriptome, and metabolome experiments. Our findings reveal that individual acute oral treatments with pesticides, specifically 20 μL of 10 ng/g imidacloprid (0.2 ng per bee), 30 ng/g chlorpyrifos (0.6 ng per bee), and 60 ng/g glyphosate (1.2 ng per bee), did not impact the flight capability of the bees. However, when bees were exposed to a combination of two or three pesticides, a notable reduction in flight duration and distance was observed. In the transcriptomic and metabolomic analyses, we identified 307 transcripts and 17 metabolites that exhibited differential expression following exposure to combined pesticides, primarily associated with metabolic pathways involved in energy regulation. Our results illuminate the intricate effects and potential hazards posed by combined pesticide exposures on bee behavior. These findings offer valuable insights into the synergistic potential of pesticide combinations and their capacity to impair bee behavior. Understanding these complex interactions is essential for comprehending the broader consequences of pesticide formulations on honey bee populations., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2024. Published by Elsevier Inc.)

Ceratitis capitata (medfly) is one of the most devastating crop pests worldwide. The Sterile Insect Technique (SIT) is a control method that is based on the mass rearing of males, their sterilization, and release in the field. However, the effectiveness of the technique depends on the quality of the released males and their fitness. We previously isolated and selected a probiotic bacteria (Enterobacter sp.), from wild-caught medflies, according to criteria that improved biological quality traits of reared medfly males.We firstly evaluated the impact of the irradiation on the expression of different immune and stress genes in the medfly sterile males. Expression was measured at differents time points ranging from 0 to 168 h after irradiation to capture the response of genes with distinct temporal expression patterns. Then, we supplemented the larval diet with previously isolated Enterobacter sp.strain, live and autoclaved at various concentrations to see whether the probiotic treatments affect, through their protective role, the gene expression level, and quality traits. The irradiation had significant effect on the genes attacin, cecropin, PGPR-LC, hsp23, and hsp70 level expression. The expression of attacin and PGPR-LC was up-regulated while that of cecropin was down-regulated. Hsp genes showed decreased levels between 0 and 18 h to peak at 72 h. However, the supplementation of the probiotic strain, either live or autoclaved, was statistically significant only for attacingene. However, significant interaction time x probiotic was noticed for attacin, cecropin, hsp23 and hsp70. The probiotic treatments also improved the quality control parameters like pupal weight. From this work we can conclude that a consortium of parabiotics (autoclaved probiotics) treatment will be recommended in insectaries considering both the beneficial effects on mass reared insects and its general safety for insectary workers and for environment., Competing Interests: The authors have declared that no competing interests exist.

Parkinson disease (PD) is a progressive, neurodegenerative disorder affecting over 6.1 million people worldwide. Although the cause of PD remains unclear, studies of highly penetrant mutations identified in early-onset familial parkinsonism have contributed to our understanding of the molecular mechanisms underlying disease pathology. Dopamine (DA) transporter (DAT) deficiency syndrome (DTDS) is a distinct type of infantile parkinsonism-dystonia that shares key clinical features with PD, including motor deficits (progressive bradykinesia, tremor, hypomimia) and altered DA neurotransmission. Here, we define structural, functional, and behavioral consequences of a Cys substitution at R445 in human DAT (hDAT R445C), identified in a patient with DTDS. We found that this R445 substitution disrupts a phylogenetically conserved intracellular (IC) network of interactions that compromise the hDAT IC gate. This is demonstrated by both Rosetta molecular modeling and fine-grained simulations using hDAT R445C, as well as EPR analysis and X-ray crystallography of the bacterial homolog leucine transporter. Notably, the disruption of this IC network of interactions supported a channel-like intermediate of hDAT and compromised hDAT function. We demonstrate that Drosophila melanogaster expressing hDAT R445C show impaired hDAT activity, which is associated with DA dysfunction in isolated brains and with abnormal behaviors monitored at high-speed time resolution. We show that hDAT R445C Drosophila exhibit motor deficits, lack of motor coordination (i.e. flight coordination) and phenotypic heterogeneity in these behaviors that is typically associated with DTDS and PD. These behaviors are linked with altered dopaminergic signaling stemming from loss of DA neurons and decreased DA availability. We rescued flight coordination with chloroquine, a lysosomal inhibitor that enhanced DAT expression in a heterologous expression system. Together, these studies shed some light on how a DTDS-linked DAT mutation underlies DA dysfunction and, possibly, clinical phenotypes shared by DTDS and PD., Competing Interests: JA, MC, JF, AS, KL, SM, AB, YZ, AC, LS, MK, CF, JM, RR, HM, IB, HM, AG No competing interests declared, AD is now employed at Cyrus Biotechnology with granted stock options. However, all contributions to the present work were made during AD's graduate education at Vanderbilt University

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) can damage dopaminergic neurons in the substantia nigra in many mammals with biochemical and cellular changes that are relatively similar to those observed in Parkinson's disease. Our study examined whether MPTP-treated echolocation bats can cause changes in bat echolocation system. By considering ultrasound spectrums, auditory brainstem-evoked potentials and flight trajectories of normal bats, we observed that the vocal, auditory, orientation and movement functions of MPTP-treated bats were significantly impaired, and they exhibited various symptoms resembling those in patients with Parkinson's disease. Our immunohistochemistry and western blot analyses further indicated that expression of vocal-related FOXP2 in the superior colliculus, auditory-related otoferlin in the inferior colliculus, dopamine synthesis-related aromatic l-amino acid decarboxylase in the substantia nigra and dopamine receptor in the striatum was significantly decreased. Furthermore, protein expression related to inflammation, oxidative stress and apoptosis in the substantia nigra was significantly increased in MPTP-treated bats. These results indicate that inflammation, oxidative stress and apoptosis may be instrumental in dopaminergic neurodegeneration in the substantia nigra. The vocal, auditory and orientation and movement dysfunctions of MPTP-treated bats are relatively consistent with symptoms of Parkinson's disease., (Copyright © 2020 The Author(s). Published by Wolters Kluwer Health, Inc.)

Rapidly increasing pyrethroid insecticide resistance and changes in vector biting and resting behavior pose serious challenges in malaria control. Mosquito repellents, especially spatial repellents, have received much attention from industry. We attempted to simulate interactions between mosquitoes and repellents using a machine learning method, the Self-Propelled Particle (SPP) model, which we modified to include attractiveness/repellency effects. We simulated a random walk scenario and scenarios with insecticide susceptible/resistant mosquitoes against repellent alone and against repellent plus attractant (to mimic a human host). Simulation results indicated that without attractant/repellent, mosquitoes would fly anywhere in the cage at random. With attractant, all mosquitoes were attracted to the source of the odor by the end. With repellent, all insecticide-susceptible mosquitoes eventually moved to the corner of the cage farthest from the repellent release point, whereas, a high proportion of highly resistant mosquitoes might reach the attractant release point (the human) earlier in the simulation. At fixed concentration, a high proportion of mosquitoes could be able to reach the host when the relative repellency efficacy (compare to attractant efficacy) was <1, whereas, no mosquitoes reached the host when the relative repellency efficacy was > 1. This result implies that repellent may not be sufficient against highly physiologically insecticide resistant mosquitoes, since very high concentrations of repellent are neither practically feasible nor cost-effective., Competing Interests: The authors have declared that no competing interests exist.

Methylmercury (MeHg) is a ubiquitous environmental contaminant and developmental toxicant known to cause a variety of persistent motor and cognitive deficits. While previous research has focused predominantly on neurotoxic MeHg effects, emerging evidence points to a myotoxic role whereby MeHg induces defects in muscle development and maintenance. A genome wide association study for developmental sensitivity to MeHg in Drosophila has revealed several conserved muscle morphogenesis candidate genes that function in an array of processes from myoblast migration and fusion to myotendinous junction (MTJ) formation and myofibrillogenesis. Here, we investigated candidates for a role in mediating MeHg disruption of muscle development by evaluating morphological and functional phenotypes of the indirect flight muscles (IFMs) in pupal and adult flies following 0, 5, 10, and 15 μM MeHg exposure via feeding at the larval stage. Developmental MeHg exposure induced a dose-dependent increase in muscle detachments (myospheres) within dorsal bundles of the IFMs, which paralleled reductions eclosion and adult flight behaviors. These effects were selectively phenocopied by altered expression of kon-tiki (kon), a chondroitin sulfate proteoglycan 4/NG2 homologue and a central component of MTJ formation. MeHg elevated kon transcript expression at a crucial window of IFM development and transgene overexpression of kon could also phenocopy myosphere phenotypes and eclosion and flight deficits. Finally, the myosphere phenotype resulting from 10 μM MeHg was partially rescued in a background of reduced kon expression using a targeted RNAi approach. Our findings implicate a component of the MTJ as a MeHg toxicity target which broaden the understanding of how motor deficits can emerge from early life MeHg exposure., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Recent studies have reported interspecific differences in how bee species respond to various stressors. Evaluating the exposure and responses of different bee species to plant protection products is considered an essential part of their risk assessment. This study was conducted to assess the impacts of thiacloprid-prochloraz mixture on buff-tailed bumblebees (Bombus terrestris) and red mason bees (Osmia bicornis) in a worst-case scenario under semi-field conditions. Bumblebee colonies or solitary bee trap nests were confined in tunnels with flowering oilseed rape. The recommended maximum application rates of 72 g thiacloprid/ha and 675 g prochloraz/ha were applied as a tank mixture during bee flight in full flowering oilseed rape. Several parameters such as flight and foraging activity, population parameters, and exposure level were investigated. Our results show adverse effects of the combination of thiacloprid and prochloraz on the reproductive performance of red mason bees. The number of cocoons produced by O. bicornis was significantly reduced in the treatment compared to the control group. Regarding bumblebees, we found no effects of the thiacloprid-prochloraz mixture on any observed parameters of colony development. The maximum detected concentrations of both active substances three days after application were higher in O. bicornis pollen mass compared to B. terrestris stored pollen. We conclude that this worst-case scenario of thiacloprid-prochloraz exposure poses a high risk to solitary bees and thus the use of such mixture should be restricted.

Trehalose is a disaccharide that is the major sugar found in insect hemolymph fluid. Trehalose provides energy, and promotes growth, metamorphosis, stress recovery, chitin synthesis, and insect flight. The hydrolysis of trehalose is under the enzymatic control of the enzyme trehalase. Trehalase is critical to the role of trehalose in insect physiology, and is required for the regulation of metabolism and glucose generation. Trehalase inhibitors represent a novel class of insecticides that have not been fully developed. Here, we tested the ability of trehalose analogues to function as larvacides or adulticides in an important disease vector-Aedes aegypti. We show that validamycin A, but not 5-thiotrehalose, delays larval and pupal development and prevents flight of adult mosquitoes. Larval mosquitoes treated with validamycin A were hypoglycemic and pupae had increased levels of trehalose. Treatment also skewed the sex ratio toward male mosquitoes. These data reveal that validamycin A is a mosquito adulticide that can impair normal development of an important disease vector., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The extensive use of insecticides can cause adverse side effects on pollinators, which negatively impact crop productivity. The pollination carried out by the honeybee Apis mellifera L. (Hymenoptera: Apidae) is crucial in increasing the productivity of the melon (Cucumis melo L.). The main objective of this study was to assess if insecticides applied in the cultivation of cantaloupe melon exhibit significant levels of toxicity toward A. mellifera. We tested the toxicity of azadirachtin, pyriproxyfen, chlorantraniliprole, and imidacloprid, which are commonly sprayed to manage melon pests such as the whitefly Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae), the pickleworm Diaphania nitidalis (Stoll) and the melonworm Diaphania hyalinata (L.) (Lepidoptera: Pyralidae). Three treatments were carried out, 0.0×, 0.1x and 1.0x the concentration recommended by the manufacturer for the control of those pests. Repellency tests, analysis of mortality through contact and ingestion, and flight tests were performed. The insecticide imidacloprid caused mortality rates above 90% in all tested exposure pathways, displaying high residue persistence on plants. Although not causing significant mortality in the ingestion test, pyriproxyfen caused significant mortality after exposure through contact, and change in flight ability. Azadirachtin caused mortality in the ingestion test and impaired the flight ability of bees, while chlorantraniliprole only impaired the flight ability. Moreover, bees were not repelled by these insecticides, suggesting that they may collect contaminated food in the field while foraging. Altogether, ecofriendly, alternative pest control options should be developed, as well as the adoption of more selective insecticides, in order to reduce the non-target effects on honeybees and guarantee their pollination services.

Background: In view of widespread pyrethroid resistance in malaria vectors in Africa, two long-lasting insecticidal nets (LLINs) incorporated with a synergist, piperonyl butoxide (PBO), DawaPlus 3.0 (deltamethrin + PBO in the roof panel; deltamethrin alone in the side panels) and DawaPlus 4.0 (deltamethrin + PBO in all panels), were evaluated in an experimental hut trial in a rice growing irrigated area in Burkina Faso. Efficacy of nets was tested against free-flying malaria vector, Anopheles gambiae s.l., with high pyrethroid resistance involving L1014F kdr and CYP6P3P450 resistance mechanisms., Methodology: The efficacy of unwashed and 20-times washed DawaPlus 3.0 (polyethylene roof panel with 120 mg/m2 deltamethrin and 440 mg/m2 PBO; polyester side panels with deltamethrin 100 mg/m2) and DawaPlus 4.0 (same composition as roof of DawaPlus 3.0) was evaluated against DawaPlus 2.0 (80 mg/m2 deltamethrin; positive control). Volunteer sleepers and treatments were rotated in huts using a Latin square design on 63 consecutive nights during August-October 2016. Mortality, human blood-feeding inhibition, deterrence and exit rates of An. gambiae s.l. were monitored., Principal Findings: Significantly higher rates of mortality and blood-feeding inhibition were observed with unwashed DawaPlus 4.0 (36%; 47.5%) than unwashed DawaPlus 3.0 (11.8%; 33.3%), DawaPlus 2.0 (4.3%; 6.4%) or untreated net (P < 0.05). Washing reduced personal protective efficacy yet PBO-LLINs were more protective and both met the WHO criteria., Conclusions: The PBO-containing DawaPlus 4.0 significantly protected against An. gambiae s.l. in the study area. Unwashed DawaPlus 3.0 gave low to moderate protection against the positive control. PBO inhibits oxidase action; hence in areas with active malaria transmission having oxidase mechanisms, PBO nets could confer additional personal protection., Competing Interests: All the authors declare that they have no competing interests.

As a managed agricultural pollinator, the western honeybee Apis mellifera frequently encounters agrochemicals as contaminants of nectar and pollen. One such contaminant, the fungicide boscalid, is applied at bloom in orchards for fungal floral pathogen control. As an inhibitor of complex II in the mitochondrial electron transport chain of fungi, boscalid can potentially interfere with high energy-demanding activities of bees, including flight. We designed an indoor flight treadmill to evaluate impacts of ingesting boscalid and/or quercetin, a ubiquitous phytochemical in bee food that also affects mitochondrial respiration. Boscalid reduced the wingbeat frequencies of foragers during flight but did not alter the duration of flight. At the colony level, boscalid ingestion may thereby affect overall health by reducing forager efficiency. The consumption of quercetin, by contrast, led to higher adenosine triphosphate levels in flight muscles and a higher wingbeat frequency. Consuming the two compounds together increased wingbeat frequency, demonstrating a hitherto unrecognized mechanism by which dietary phytochemicals may act to ameliorate toxic effects of pesticides to promote honeybee health. In carrying out this work, we also introduce two methodological improvements for use in testing for pesticide effects on flight capacity-a 'force-feeding' to standardize flight fuel supply and a novel indoor flight treadmill.

Flupyradifurone (FPF, Sivanto®) is a new butenolide insecticide that, like the neonicotinoids, is a systemic nicotinic acetylcholine receptor (nAChR) agonist. However, FPF is considered bee-safe (according to standard Risk Assessment tests), and is thus a potential solution to the adverse effects of other pesticides on beneficial insects. To date, no studies have examined the impact of nutritional stress (decreased food diversity and quality) and FPF exposure on bee health although both stressors can occur, especially around agricultural monocultures. We therefore tested the effects of a field-realistic FPF concentration (4 ppm, FPF daily dose  = 241 ± 4 ng/bee/day, 1/12 of LD 50 ) and nutritional stress (nectar with low-sugar concentrations) on honey bee (Apis mellifera L.) mortality, food consumption, thermoregulation, flight success (unsuccessful vs. successful), and flight ability (duration, distance, velocity). Flight and thermoregulation are critical to colony health: bees fly to collect food and reproduce, and they thermoregulate to increase flight efficiency and to rear brood. We studied the effects across seasons because seasonality can influence bee sensitivity to environmental stress. We demonstrate that, depending upon season and nutritional stress, FPF can reduce bee survival (-14%), food consumption (-14%), thermoregulation (-4%, i.e. hypothermia), flight success (-19%), and increase flight velocity (+13%). Because pesticide exposure and nutritional stress can co-occur, we suggest that future studies and pesticide risk assessments consider both seasonality and nutritional stress when evaluating pesticide safety for bees., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

During migratory flight, desert locusts rely on fatty acids as their predominant source of energy. Lipids mobilized in the fat body are transported to the flight muscles and enter the muscle cells as free fatty acids. It has been postulated that muscle fatty acid binding protein (FABP) is needed for the efficient translocation of fatty acids through the aqueous cytosol towards mitochondrial β-oxidation. To assess whether FABP is required for this process, dsRNA was injected into freshly emerged adult males to knock down the expression of FABP. Three weeks after injection, FABP and its mRNA were undetectable in flight muscle, indicating efficient silencing of FABP expression. At rest, control and treated animals exhibited no morphological or behavioral differences. In tethered flight experiments, both control and treated insects were able to fly continually in the initial, carbohydrate-fueled phase of flight, and in both groups, lipids were mobilized and released into the hemolymph. Flight periods exceeding 30 min, however, when fatty acids become the main energy source, were rarely possible for FABP-depleted animals, while control insects continued to fly for more than 2 h. These results demonstrate that FABP is an essential element of skeletal muscle energy metabolism in vivo ., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Background: Thiamethoxam (TMX) represents the second generation of neonicotinoids that has been widely applied in agricultural activities, while how TMX alters the behavior of Apis cerana, an important native honey bee species in China, is not clear. We carried out three independent experiments to study the impact of acute oral treatment of 20 μL TMX at concentrations of 2.4 ppb (0.048 ng/bee) and 10 ppb (0.2 ng/bee) on the homing, flight, learning acquisition and short-term retention ability of A. cerana. The homing ability was assessed by the catch-and-release method, the flight ability was assessed by flight mills, and the learning acquisition and short-term retention were evaluated by the proboscis extension response method., Results: When treated with 10 ppb of TMX, bees had a significantly higher average homing time, mean flight velocity, flying distance, and flying duration than the control, whereas 2.4 ppb concentration did not cause any significant effect on homing or flight ability. Bees treated with either 2.4 ppb or 10 ppb TMX had significantly lower learning acquisition and short-term retention ability., Conclusion: Results suggest that acute oral exposure to 10 ppb of TMX altered the short-distance homing time, flight ability, and learning acquisition and short-term retention ability. Our study also highlights the concern that acute oral exposure to a low concentration of 2.4 ppb could have consequences on the behavior of A. creana. Those multiple sublethal alterations on A. cerana's behavior indicate that TMX are likely having complex but negative consequences on bee health in the field. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

The BG-Malaria trap (BGM) is an adaptation of the well-known BG-Sentinel trap (BGS) with greater trapping efficiencies for anopheline and culicine mosquitoes. Its continued optimization requires greater understanding of mosquito flight behaviors near it. We used three high-resolution infrared cameras (68 frames/second) to track flight behaviors of laboratory-reared Anopheles arabiensis females in vicinity of the BGM in comparison with BGS. Additional comparisons were done for BGM at 20, 40 and 80cm heights, and for BGMs baited with Ifakara blend plus CO2, CO2 alone, or no bait. More mosquitoes were observed near BGM than BGS. Both BGMs installed 20cm above the floor and baited with CO2 received more visits by host-seeking mosquitoes than the other BGMs evaluated in their respective experiments. Trap designs, height and attractants all influence mosquito activity in vicinity of the traps which can be readily visualized using infrared cameras to accelerate trap development and testing. The greater activity of host-seeking mosquitoes near BGM than BGS supports the proven superiority of BGM traps in field and semi-field settings., Competing Interests: The authors have declared that no competing interests exist.

While neonicotinoid insecticides impair visually guided behaviours, the effects of their metabolites are unknown and measurements of environmental concentrations of neonicotinoids, typically lower than those required to elicit toxic effects, tend to exclude metabolites. Here we examined the contributions of imidacloprid and two of its metabolites, imidacloprid-olefin and 5-hydroxy-imidacloprid, on neural conduction velocity, visual motion detection and flight in the locust (Locusta migratoria) using a combination of electrophysiological and behavioural assays. We show reduced visual motion detection and impaired flight behaviour following treatment of metabolite concentrations equal to sublethal doses of the parent compound. Additionally, we show for the first time that imidacloprid and its metabolites result in a decrease in conduction velocity along an unmyelinated axon. We suggest that secondary effects of the insecticide on the biophysical properties of the axon may result in decreased neural conduction. As these metabolites display neurotoxicity similar to the parent compound they should be considered when quantifying environmental concentrations., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The migratory flights of birds are primarily fueled by fat; however, certain fatty acids may also enhance flight performance and the capacity to oxidize fat. The natural doping hypothesis posits that n-3 long-chain polyunsaturated fatty acids (PUFA) increase membrane fluidity and aerobic and fatty acid oxidative enzymes in the flight muscles, which enables prolonged endurance flight. Support for this hypothesis is mixed, and there is no empirical evidence for increased flight performance. We fed yellow-rumped warblers ( Setophaga coronata coronata) diets enriched in either n-3 or n-6 long-chain PUFA or low in long-chain PUFA and evaluated flight muscle metabolism and endurance performance in a wind tunnel flights lasting up to 6 h. Fatty acid profiles of muscle phospholipids confirmed enrichment of the targeted dietary fatty acids, whereas less substantial differences were observed in adipose triacylglycerol. Contrary to the predictions, feeding n-3 PUFA decreased peroxisome proliferator-activated receptors-β mRNA abundance and muscle oxidative enzyme activities. However, changes in muscle metabolism were not reflected in whole animal performance. No differences were observed in flight performance among diet treatments in terms of endurance capacity, energy costs, or fuel composition. These measures of flight performance were more strongly influenced by body mass and flight duration. Overall, we found no support for the natural doping hypothesis in a songbird. Furthermore, we caution against extending changes in flight muscle metabolic enzymes or fatty acid composition to changes to migratory performance without empirical evidence.

Aedes aegypti and Anopheles gambiae harbor the causative agents of diseases such as dengue fever and malaria, afflicting human morbidity and mortality worldwide. Given the worldwide emergence of resistance to insecticides, the current mainstay for vector control, identification of alternative modes of action for future insecticides is paramount. The serotonergic (5-HT) system has been documented to impact physiological mechanisms involved in disease transmission, suggesting its potential as a new mode of action target for future insecticide development. Target 5-HT receptors were cloned and expressed in the HEK293 cell line for functional and pharmacological characterization. Manipulation of the 5-HT system through microinjection of compounds suggests its involvement in the modulation of flight performance and blood-feeding behavior. By attenuating these two determinants of vectorial capacity, transmission and burden of disease could effectively be reduced. Considering these positive global health implications, the 5-HT system is a compelling target for the novel insecticide pipeline.

The current-use brominated flame retardant, 1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane (DBE-DBCH), is capable of perturbing sex steroid and thyroid hormone pathways in vitro and in vivo. Chemicals with this capability may also disrupt metabolic processes and are candidate obesogens, but this potential has not yet been determined for DBE-DBCH. Our objective was to examine gross biomarkers of metabolic disruption in captive American kestrels. Birds were exposed by diet to the β isomer at the environmentally relevant dose of 0.239 ng β-DBE-DBCH/g kestrel/day, from 30 days (d) prior to pairing through until chicks hatched (82 d) (n = 30 breeding pairs) or for 28 d (n = 16 pre-breeding pairs), and were compared with vehicle-only exposed controls. Body mass was assessed throughout the breeding season at biologically relevant time points, flight and feeding behavior was measured in 5-min samples daily, and plasma triglycerides and cholesterol were assessed at d10 of brood rearing. Treated males were heavier than controls at pairing (p = 0.051), the final week of courtship (p = 0.061), and at d10 (p = 0.012) and d20 of brood rearing (p = 0.051); β-DBE-DBCH-exposed breeding females were similar in weight to control females. Treated birds tended to have higher plasma triglycerides (p = 0.078), which for females, was positively associated with body mass (p = 0.019). Heavier breeding males had higher plasma concentrations of testosterone and total thyroxine (p ≤ 0.046). Overall, both sexes exposed to β-DBE-DBCH demonstrated reduced flight behavior and increased feeding behavior during courtship. In the pre-breeding pairs, treated male and female kestrels had a higher percentage of body fat than respective controls (p = 0.045). These results demonstrate that β-DBE-DBCH elicited inappropriate fat and weight gain in adult American kestrels, consistent with their increased feeding, reduced flight activity and endocrine changes, and suggests that DBE-DBCH may be an obesogen warranting further research to test this hypothesis., (Crown Copyright © 2018. Published by Elsevier Inc. All rights reserved.)

A semifield study to assess the effects of iprodione on honeybees at label use rates was conducted on a bloom mustard crop. The present study followed the Organisation for Economic Co-operation and Development guideline 75 tunnel test and consisted of 3 groups: the iprodione-treated group, the untreated control group, and the toxic reference item group. In addition to the tunnels used for biological assessments, a tunnel was set up in the treatment and control groups to determine the level of residues in flowers, nectar, and pollen. The major endpoints to assess the effects of the application of iprodione were mortality, flight intensity, behavior, condition of the colonies, and development of the brood. Residue analysis showed that honeybees were exposed to significant residues of iprodione. However, no adverse effects were observed on overall mortality, flight intensity, behavior, or brood development of honeybees compared to control. It is concluded that iprodione does not adversely affect the health of honeybees when applied in agriculture at commercially relevant rates in a worst-case exposure scenario. Environ Toxicol Chem 2018;37:3086-3094. © 2018 SETAC., (© 2018 SETAC.)

Myclobutanil is currently used on the flowering plants. Little is known about how Apis cerana cerana respond to myclobutanil exposure. Hence, the acute toxicity of myclobutanil and its sublethal effects on respiration, flight and detoxification enzymes [7-ethoxycoumarin O-deethylase (ECOD) and glutathione S-transferases (GSTs)] in A. cerana cerana were investigated. The results indicated that formulation grade myclobutanil showed moderate toxicity to A. cerana cerana either contact (LD 50 =4.697μg/bee) or oral (LD 50 =2.154μg/bee) exposure. Sublethal dose of myclobutanil significantly reduced the respiration rate of workers at 24h and 48h regardless of the exposure method. However, myclobutanil didn't significantly affect the take-off flight. After nurse bees exposure to the dose (LD 5 ) of formulation-grade myclobutanil, ECOD activity was significantly induced when compared with control, but GST activity didn't change. In the forager bees, no enzyme markers response was obtained in this test. From the present study we can infer that myclobutanil disturb respiration and P450-mediated detoxification of the individual bees of A. cerana cerana. Thus, myclobutanil may has risk for A. cerana cerana, it should be cautiously used., (Copyright © 2017. Published by Elsevier Inc.)

The chemical agent sulfur mustard (SM) causes erythema, skin blisters, ulcerations, and delayed wound healing. It is accepted that the underlying molecular toxicology is based on DNA alkylation. With an expected delay, DNA damage causes impairment of protein biosynthesis and disturbance of cell division. However, using the cockroach model Blaptica dubia, the presented results show that alkylating compounds provoke immediate behavior responses along with fast changes in the electrical field potential (EFP) of neurons, suggesting that lesions of DNA are probably not the only effect of alkylating compounds. Blaptica dubia was challenged with SM or 2-chloroethyl-ethyl sulfide (CEES). Acute toxicity was objectified by a disability score. Physiological behavior responses (antennae pullback reflex, escape attempts, and grooming) were monitored after exposure. To estimate the impact of alkylating agents on neuronal activity, EFP recordings of the antennae and the thoracic ganglion were performed. After contact to neat SM, a pullback reflex of the antennae was the first observation. Subsequently, a striking escape behavior occured which was characterized by persistent movement of the legs. In addition, an instantaneous processing of the electrical firing pattern from the antennae to the descending ganglia was detectable. Remarkably, comparing the toxicity of the applied alkylating agents, effects induced by CEES were much more pronounced compared to SM. In summary, our findings document immediate effects of B. dubia after exposure to alkylating substances. These fast responses cannot be interpreted as a consequence of DNA alkylation. Therefore, the dogma that DNA alkylation is the exclusive cause for SM toxicity has to be questioned.

In 2010, the Deepwater Horizon oil spill released 134 million gallons of crude oil into the Gulf of Mexico making it the largest oil spill in US history. The three month oil spill left tens of thousands of birds dead; however, the fate of tens of thousands of other migratory birds that were affected but did not immediately die is unknown. We used the homing pigeon as a surrogate species for migratory birds to investigate the effects of a single external oiling event on the flight performance of birds. Data from GPS data loggers revealed that lightly oiled pigeons took significantly longer to return home and spent more time stopped en route than unoiled birds. This suggests that migratory birds affected by the oil spill could have experienced long term flight impairment and delayed arrival to breeding, wintering, or crucial stopover sites and subsequently suffered reductions in survival and reproductive success., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The Deepwater Horizon oil spill contaminated thousands of miles of habitat valuable to hundreds of species of migratory and resident birds of the Gulf of Mexico. Many birds died as a direct result of the oil spill; however, the indirect effects of oil exposure on the flight ability and body condition of birds are difficult to assess in situ. This study utilizes the homing pigeon as a surrogate species for migratory birds to investigate the effect of multiple external oil exposures on the flight performance and body mass change of birds over a series of repeated flights from 136.8km flight distance. Oiled pigeons took significantly longer to return home, lost more weight during flight, and were unable to recover their weight, resulting in reduction of body weight overtime. Based on our data, migratory birds that were oiled, even partially, by the Deepwater Horizon oil spill likely took longer to complete migration and were likely in poor body condition, increasing their risk of mortality and reproductive failure., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The Deepwater Horizon oil spill was the largest in U.S. history, contaminating thousands of miles of coastal habitat and affecting the lives of many avian species. The Gulf of Mexico is a critical bird migration route area and migrants that were oiled but did not suffer mortality as a direct result of the spill faced unpredictable fates. This study utilized homing pigeons as a surrogate species for migratory birds to investigate the effects a single low level external oiling event has on the flight performance and behavior of birds flying repeated 161 km flights. Data from GPS data loggers showed that lightly oiled pigeons changed their flight paths, increased their flight durations by 2.6 fold, increased their flight distances by 28 km and subsequently decreased their route efficiencies. Oiled birds also exhibited reduced rate of weight gain between flights. Our data suggest that contaminated birds surviving the oil spill may have experienced flight impairment and reduced refueling abilities, likely reducing overall migration speed. Our findings contribute new information on how oil spills affect avian species, as the effects of oil on the flight behavior of long distance free-flying birds have not been previously described., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Fluorescent dyes are commonly used in the sterile insect technique (SIT) for marking insects for a proper identification after recapture. However, the quality of the mark must be balanced against insect performance, because dyes can negatively affect some parameters of insect performance and reduce their effectiveness in control with the SIT. We determined the visibility and persistence and the effect of dyes on the quality of Anastrepha obliqua (Macquart) and Anastrepha ludens (Loew) (bisexual and genetic sexing strains) by testing four concentrations of a dye (Day-Glo) from 0 to 2.5 g dye/kg of pupae. Visibility and persistence of the mark were positively affected by dose and negatively affected by the length of time the samples were kept in a solution of 75% alcohol. However, upon dissection, even the lowest dose of dye was visible under a fluorescence microscope. Between dyed and undyed pupae (control), no significant differences were observed in rates of emergence, fliers and flight ability, and survival in two tests, with water and without food and without water and food, at any of the concentrations tested. Furthermore, no significant difference in mating competitiveness was detected between control pupae and those dyed at 1.0 and 2.5 g dye/kg pupae. We discuss our results with the possibility of reducing the dose of dye in these three flies, because the heads are large enough to capture sufficient particles to permit identification with the current methods of detection., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Insecticide-treated nets are currently a major tool to reduce malaria transmission. Their level of repellency affects contact of the mosquito with the net, but may also influence the mosquito's entry into the house. The response of host-seeking malaria mosquitoes approaching the eave of an experimental house was recorded within a large screen house. We compared entry- and exit rates in relation to the presence in the house of different insecticide-treated bed nets (ITNs) with an untreated net. Mosquitoes were lured towards the house by dispensing a synthetic host-odour blend from within the net in the house. Complementary WHO bioassays revealed that the treated nets caused high knock-down- and mortality responses to the Anopheles gambiae sensu stricto strain tested. The proportion of mosquitoes that came into view of the cameras and subsequently entered the house did not differ between treated nets and the untreated net. Treated nets did not affect proportions of mosquitoes that exited the house and departed from view around the eave. However, the percentage of house-leaving and re-entering mosquitoes when an insecticide- treated net was present, was lower than in the presence of an untreated net. Our results indicated that there was no spatial repellent effect from pyrethroid-treated nets that influences house-entry at eave level. It is argued that the toxic effect of treated bed nets resulted in a reduced number of mosquitoes re-entering the house, which could thereby affect malaria transmission in neighbouring, unprotected houses., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

The ability to takeoff quickly and accelerate away from predators is crucial to bird survival. Crude oil can disrupt the fine structure and function of feathers, and here we tested for the first time how small amounts of oil on the trailing edges of the wings and tail of Western sandpipers (Calidris mauri) affected takeoff flight performance. In oiled birds, the distance travelled during the first 0.4s after takeoff was reduced by 29%, and takeoff angle was decreased by 10° compared to unoiled birds. Three-axis accelerometry indicated that oiled sandpipers produced less mechanical power output per wingbeat during the initial phase of flight. Slower and lower takeoff would make oiled birds more likely to be targeted and captured by predators, reducing survival and facilitating the exposure of predators to oil. Whereas the direct mortality of heavily-oiled birds is often obvious and can be quantified, our results show that there are significant sub-lethal effects of small amounts crude oil on feathers, which must be considered in natural resource injury assessments for birds., (Copyright © 2017 Elsevier Inc. All rights reserved.)

To understand the effects of pollutants in a changing world we need multistressor studies that combine pollutants with other stressors associated with global change such as heat waves. We tested for the delayed and combined impact of a heat wave during the egg stage and subsequent sublethal exposure to the pesticide esfenvalerate during the larval stage on life history and physiology in the larval and adult stage of the damselfly Lestes sponsa. We studied this in a common garden experiment with replicated central- and high latitude populations to explore potential effects of local thermal adaptation and differences in life history shaping the multistressor responses. Exposure of eggs to the heat wave had no effect on larval traits, yet had delayed costs (lower fat and flight muscle mass) in the adult stage thereby crossing two life history transitions. These delayed costs were only present in central-latitude populations potentially indicating their lower heat tolerance. Exposure of larvae to the pesticide reduced larval growth rate and prolonged development time, and across metamorphosis reduced the adult fat content and the flight muscle mass, yet did not affect the adult heat tolerance. The pesticide-induced delayed emergence was only present in the slower growing central-latitude larvae, possibly reflecting stronger selection to keep development fast in the more time-constrained high-latitude populations. We observed no synergistic interactions between the egg heat wave and the larval pesticide exposure. Instead the pesticide-induced reduction in fat content was only present in animals that were not exposed to the egg heat wave. Our results based on laboratory conditions highlight that multistressor studies should integrate across life stages to fully capture cumulative effects of pollutants with other stressors related to global change., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Pesticides can pose environmental risks, and a common neonicotinoid pesticide, thiamethoxam, decreases homing success in honey bees. Neonicotinoids can alter bee navigation, but we present the first evidence that neonicotinoid exposure alone can impair the physical ability of bees to fly. We tested the effects of acute or chronic exposure to thiamethoxam on the flight ability of foragers in flight mills. Within 1 h of consuming a single sublethal dose (1.34 ng/bee), foragers showed excitation and significantly increased flight duration (+78%) and distance (+72%). Chronic exposure significantly decreased flight duration (-54%), distance (-56%), and average velocity (-7%) after either one or two days of continuous exposure that resulted in bees ingesting field-relevant thiamethoxam doses of 1.96-2.90 ng/bee/day. These results provide the first demonstration that acute or chronic exposure to a neonicotinoid alone can significantly alter bee flight. Such exposure may impair foraging and homing, which are vital to normal colony function and ecosystem services.

Sogatella furcifera Horváth (Hemiptera: Delphacidae), is a major migratory pest of rice crops in Asia. The ultrastructure of the flight muscle directly affects the flight ability of insects. The ultrastructure of the flight muscle of some insects can be affected by insecticides. However, the ultrastructure of the flight muscle of S. furcifera and the effect of insecticides on the flight muscle of S. furcifera are not well understood. The present study was conducted to determine the effect of the insecticide dinotefuran on the ultrastructure of the flight muscle of S. furcifera females. In this study, the cross-sectional area and the diameter of the myofibril cross-sections of dinotefuran-treated S. furcifera females increased with the number of days after emergence (DAE), and they were higher than in untreated females. The sarcomere length of myofibrils increased with the number of DAE, and it differed from that of the untreated females. On the first day after emergence, the higher the concentration of dinotefuran, the smaller was the extent of decrease. On the third day after emergence, the higher the concentration of dinotefuran, the larger was the extent of enhancement. For the percentage of mitochondria, those of LC10 and LC20 dinotefuran-treated S. furcifera females increased with the number of DAE and were higher than in untreated females. LC10 dinotefuran-treated S. furcifera females exhibited the largest increase. Thus, our results suggest that the flight ability of S. furcifera increased with time. Some concentrations of dinotefuran can enhance the flight capacity of S. furcifera., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: The direct lethal effects of conventional and organic insecticides have been investigated thoroughly for all life stages of Halyomorpha halys. However, the sublethal effects of insecticides on the behavior of H. halys have not been well documented. Our aims were to evaluate the impact of a brief 5 min exposure to residues of bifenthrin, dinotefuran, methomyl, thiamethoxam and thiamethoxam + λ-cyhalothrin on survivorship, horizontal and vertical movement, and flight capacity of adult H. halys under laboratory conditions., Results: Over half of the insecticide-exposed adults were classified as affected, moribund or dead after the 5 min exposure, compared with only 6% of the adults in the water-only control. We found that the horizontal movement, vertical climbing and flight capacity of adults exposed to insecticides were decreased by 20-60% overall relative to the water-only control. The most lethal insecticide was bifenthrin., Conclusion: Many insecticide-exposed H. halys adults retained significant mobility and flight capacity, with flight most pronounced immediately after exposure. These results suggest that brief exposure periods to efficacious insecticides will result in high dispersal and low mortality. Therefore, management strategies that enhance the retention of H. halys on insecticide-coated surfaces should be considered to ensure that adults are exposed to a lethal dose of insecticide. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

A mixture of behaviorally active volatiles was identified from the fruit of snowberry, Symphoricarpos albus laevigatus, for Rhagoletis zephyria flies reared from snowberry fruit. A nine-component blend containing 3-methylbutan-1-ol (3%), dimethyl trisulfide (1%), 1-octen-3-ol (40%), myrcene (8%), nonanal (9%), linalool (13%), (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT, 6%), decanal (15%), and β-caryophyllene (5%) was identified that gave consistent electroantennogram activity and was behaviorally active in flight tunnel tests. In other flight tunnel assays, snowberry flies from two sites in Washington state, USA, displayed significantly greater levels of upwind oriented flight to sources with the snowberry volatile blend compared with previously identified volatile blends from domestic apple (Malus domestica) and downy hawthorn (Crataegus mollis) fruit from the eastern USA, and domestic apple, black hawthorn (C. douglasii) and ornamental hawthorn (C. monogyna) from Washington state. Selected subtraction assays showed that whereas removal of DMNT or 1-octen-3-ol significantly reduced the level of upwind flight, removal of myrcene and β-caryophyllene, or dimethyl trisulfide alone did not significantly affect the proportion of upwind flights. Our findings add to previous studies showing that populations of Rhagoletis flies infesting different host fruit are attracted to unique mixtures of volatile compounds specific to their respective host plants. Taken together, the results support the hypothesis that differences among flies in their behavioral responses to host fruit odors represent key adaptations involved in sympatric host plant shifts, contributing to host specific mating and generating prezygotic reproductive isolation among members of the R. pomonella sibling species complex.

Physiological challenges during one part of the annual cycle can carry over and affect performance at a subsequent phase, and antioxidants could be one mediator of trade-offs between phases. We performed a controlled experiment with zebra finches to examine how songbirds use nutrition to manage trade-offs in antioxidant allocation between endurance flight and subsequent reproduction. Our treatment groups included (1) a non-supplemented, non-exercised group (control group) fed a standard diet with no exercise beyond that experienced during normal activity in an aviary; (2) a supplemented non-exercised group fed a water- and lipid-soluble antioxidant-supplemented diet with no exercise; (3) a non-supplemented exercised group fed a standard diet and trained to perform daily endurance flight for 6 weeks; and (4) a supplemented exercised group fed an antioxidant-supplemented diet and trained to perform daily flight for 6 weeks. After flight training, birds were paired within treatment groups for breeding. We analyzed eggs for lutein and vitamin E concentrations and the plasma of parents throughout the experiment for non-enzymatic antioxidant capacity and oxidative damage. Exercised birds had higher oxidative damage levels than non-exercised birds after flight training, despite supplementation with dietary antioxidants. Supplementation with water-soluble antioxidants decreased the deposition of lipid-soluble antioxidants into eggs and decreased yolk size. Flight exercise also lowered deposition of lutein, but not vitamin E, to eggs. These findings have important implications for future studies of wild birds during migration and other oxidative challenges., (© 2016. Published by The Company of Biologists Ltd.)

Flying insects have the highest known mass-specific demand for oxygen, which makes it likely that reduced availability of oxygen might limit sustained flight, either instead of or in addition to the limitation due to metabolite resources. The Glanville fritillary butterfly (Melitaea cinxia) occurs as a large metapopulation in which adult butterflies frequently disperse between small local populations. Here, we examine how the interaction between oxygen availability and fuel use affects flight performance in the Glanville fritillary. Individuals were flown under either normoxic (21 kPa O2) or hypoxic (10 kPa O2) conditions and their flight metabolism was measured. To determine resource use, levels of circulating glucose, trehalose and whole-body triglyceride were recorded after flight. Flight performance was significantly reduced in hypoxic conditions. When flown under normoxic conditions, we observed a positive correlation among individuals between post-flight circulating trehalose levels and flight metabolic rate, suggesting that low levels of circulating trehalose constrains flight metabolism. To test this hypothesis experimentally, we measured the flight metabolic rate of individuals injected with a trehalase inhibitor. In support of the hypothesis, experimental butterflies showed significantly reduced flight metabolic rate, but not resting metabolic rate, in comparison to control individuals. By contrast, under hypoxia there was no relationship between trehalose and flight metabolic rate. Additionally, in this case, flight metabolic rate was reduced in spite of circulating trehalose levels that were high enough to support high flight metabolic rate under normoxic conditions. These results demonstrate a significant interaction between oxygen and energy availability for the control of flight performance., (© 2016. Published by The Company of Biologists Ltd.)

Plants damaged by herbivores emit volatile organic compounds (VOCs) that are used by parasitoids for host location. In nature, however, plants are exposed to multiple abiotic and biotic stresses of varying intensities, which may affect tritrophic interactions. Here, we studied the effects of ozone exposure and feeding by Pieris brassicae larvae on the VOCs emitted by Brassica nigra and the effects on oriented flight of the parasitoid Cotesia glomerata. We also investigated the oriented flight of C. glomerata in a wind-tunnel with elevated ozone levels. Herbivore-feeding induced the emission of several VOCs, while ozone alone had no significant effect. However, exposure to 120 ppb ozone, followed by 24 hr of herbivore-feeding, induced higher emissions of all VOCs as compared to herbivore-feeding alone. In accordance, herbivore-damaged plants elicited more oriented flights than undamaged plants, whereas plants exposed to 120 ppb ozone and 24 hr of herbivore-feeding elicited more oriented flights than plants subjected to herbivore-feeding alone. Ozone enrichment of the wind-tunnel air appeared to negatively affect orientation of parasitoids at 70 ppb, but not at 120 ppb. These results suggest that the combination of ozone and P. brassicae-feeding modulates VOC emissions, which significantly influence foraging efficiency of C. glomerata.

Background: The oriental fruit fly Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) causes serious damage that affects fruit production. Chemical insecticides have been widely used for the prevention and control of this destructive pest. However, the resistance of B. dorsalis to these compounds has become a serious problem. This study tested six sweeteners for their toxicity to B. dorsalis., Results: B. dorsalis fed on erythritol, aspartame and saccharin exhibited significantly higher mortality than those fed on sucrose. Flies fed on erythritol died faster than did the control flies (water). However, no dose-dependent effects were observed at the concentrations tested. These three sweeteners decreased the climbing ability of B. dorsalis. Notably, adults fed on saccharin exhibited significantly decreased climbing ability after 12 h compared with those fed on sucrose. Additionally, these three sweeteners had a negative effect on the frequency and duration of the flies' behaviour patterns (flying, walking, grooming and inactivity). Saccharin not only induced a marked reduction in the frequency of flights and walks but also induced decreases in the time spent flying and walking and increases in inactivity compared with sucrose. Erythritol induced a reduction in movement and increased the time spent inactive compared with the control and other treatments., Conclusion: Three sweeteners had significant negative effects on the survival of B. dorsalis. Erythritol was toxic to B. dorsalis. Aspartame and saccharin also decreased the survival and behaviour of adult flies and may be toxic to (or contribute to poor nutrition in) B. dorsalis. These sweeteners could therefore be developed as additive ingredients in baits., (© 2015 Society of Chemical Industry.)

Background: Since 2010 there has been a resurgence of the saddle gall midge, Haplodiplosis marginata (von Roser), in Belgium and several other European countries, with this pest sometimes causing severe damage in cereals. In 2012 and 2013, field trials were conducted in heavily infested fields to assess its impact on winter wheat crops and to determine efficient ways of dealing with severe infestations., Results: Crop exposure to H. marginata varied with the different protection methods tried. These methods included 1-4 successive applications of lambda-cyhalothrin. Yield losses were significant, reaching 6% in 2012 and as high as 15% in 2013, and these losses were linearly related to the number of galls on stems., Conclusion: The trials showed that insecticide applications needed to be synchronised with H. marginata flight peaks and to target the egg hatching period. They also revealed that insecticides applied to coincide with the first flight could, in humid conditions, also reach the larvae close to the soil surface, prior to their pupation., (© 2015 Society of Chemical Industry.)

Honey bees and other pollinators are exposed to fungicides that act by inhibiting fungal mitochondria. Here we test whether a common fungicide (Pristine®) inhibits the function of mitochondria of honeybees, and whether consumption of ecologically-realistic concentrations can cause negative effects on the mitochondria of flight muscles, or the capability for flight, as judged by CO2 emission rates and thorax temperatures during flight. Direct exposure of mitochondria to Pristine® levels above 5 ppm strongly inhibited mitochondrial oxidation rates in vitro. However, bees that consumed pollen containing Pristine® at ecologically-realistic concentrations (≈ 1 ppm) had normal flight CO2 emission rates and thorax temperatures. Mitochondria isolated from the flight muscles of the Pristine®-consuming bees had higher state 3 oxygen consumption rates than control bees, suggesting that possibly Pristine®-consumption caused compensatory changes in mitochondria. It is likely that the lack of a strong functional effect of Pristine®-consumption on flight performance and the in vitro function of flight muscle mitochondria results from maintenance of Pristine® levels in the flight muscles at much lower levels than occur in the food, probably due to metabolism and detoxification. As Pristine® has been shown to negatively affect feeding rates and protein digestion of honey bees, it is plausible that Pristine® consumption negatively affects gut wall function (where mitochondria may be exposed to higher concentrations of Pristine®)., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Plant essential oils have been suggested as a promising alternative to the established mosquito repellent DEET (N,N-diethyl-meta-toluamide). Searching for an assay with generally available equipment, we designed a new audiovisual assay of repellent activity against mosquitoes "Singing in the Tube," testing single mosquitoes in Drosophila cultivation tubes. Statistics with regression analysis should compensate for limitations of simple hardware. The assay was established with female Culex pipiens mosquitoes in 60 experiments, 120-h audio recording, and 2580 estimations of the distance between mosquito sitting position and the chemical. Correlations between parameters of sitting position, flight activity pattern, and flight tone spectrum were analyzed. Regression analysis of psycho-acoustic data of audio files (dB[A]) used a squared and modified sinus function determining wing beat frequency WBF ± SD (357 ± 47 Hz). Application of logistic regression defined the repelling velocity constant. The repelling velocity constant showed a decreasing order of efficiency of plant essential oils: rosemary (Rosmarinus officinalis), eucalyptus (Eucalyptus globulus), lavender (Lavandula angustifolia), citronella (Cymbopogon nardus), tea tree (Melaleuca alternifolia), clove (Syzygium aromaticum), lemon (Citrus limon), patchouli (Pogostemon cablin), DEET, cedar wood (Cedrus atlantica). In conclusion, we suggest (1) disease vector control (e.g., impregnation of bed nets) by eight plant essential oils with repelling velocity superior to DEET, (2) simple mosquito repellency testing in Drosophila cultivation tubes, (3) automated approaches and room surveillance by generally available audio equipment (dB[A]: ISO standard 226), and (4) quantification of repellent activity by parameters of the audiovisual assay defined by correlation and regression analyses.

Current high losses of honeybees seriously threaten crop pollination. Whereas parasite exposure is acknowledged as an important cause of these losses, the role of insecticides is controversial. Parasites and neonicotinoid insecticides reduce homing success of foragers (e.g. by reduced orientation), but it is unknown whether they negatively affect flight capacity. We investigated how exposing colonies to the parasitic mite Varroa destructor and the neonicotinoid insecticide imidacloprid affect flight capacity of foragers. Flight distance, time and speed of foragers were measured in flight mills to assess the relative and interactive effects of high V. destructor load and a field-realistic, chronic sub-lethal dose of imidacloprid. Foragers from colonies exposed to high levels of V. destructor flew shorter distances, with a larger effect when also exposed to imidacloprid. Bee body mass partly explained our results as bees were heavier when exposed to these stressors, possibly due to an earlier onset of foraging. Our findings contribute to understanding of interacting stressors that can explain colony losses. Reduced flight capacity decreases the food-collecting ability of honeybees and may hamper the use of precocious foraging as a coping mechanism during colony (nutritional) stress. Ineffective coping mechanisms may lead to destructive cascading effects and subsequent colony collapse., (© 2015 The Author(s).)

Glyphosate (GLY) is a herbicide that is widely used in agriculture for weed control. Although reports about the impact of GLY in snails, crustaceans and amphibians exist, few studies have investigated its sublethal effects in non-target organisms such as the honeybee Apis mellifera, the main pollen vector in commercial crops. Here, we tested whether exposure to three sublethal concentrations of GLY (2.5, 5 and 10 mg l(-1): corresponding to 0.125, 0.250 and 0.500 μg per animal) affects the homeward flight path of honeybees in an open field. We performed an experiment in which forager honeybees were trained to an artificial feeder, and then captured, fed with sugar solution containing traces of GLY and released from a novel site either once or twice. Their homeward trajectories were tracked using harmonic radar technology. We found that honeybees that had been fed with solution containing 10 mg l(-1) GLY spent more time performing homeward flights than control bees or bees treated with lower concentrations. They also performed more indirect homing flights. Moreover, the proportion of direct homeward flights performed after a second release from the same site increased in control bees but not in treated bees. These results suggest that, in honeybees, exposure to levels of GLY commonly found in agricultural settings impairs the cognitive capacities needed to retrieve and integrate spatial information for a successful return to the hive. Therefore, honeybee navigation is affected by ingesting traces of the most widely used herbicide worldwide, with potential long-term negative consequences for colony foraging success., (© 2015. Published by The Company of Biologists Ltd.)

Understanding the interaction between dietary protein deficits and neurotoxicants such as lead (Pb) is critical since oxidative stress is a common denominator under such conditions. The Drosophila system is an extensively used model to investigate the interaction between nutrients and environmental toxicants. Accordingly, we have examined the hypothesis that casein (CSN) enrichment has the propensity to attenuate Pb-associated phenotype, oxidative stress and neurotoxicity in Drosophila melanogaster. Exposure of young (2-3 d) and adult flies (10-12 d old) to Pb acetate (0-20 mM, 7 d) in the medium resulted in a concentration dependent mortality and the survivors exhibited a hyperactive phenotype. While males showed higher susceptibility to Pb among both age groups, young flies were relatively more susceptible than adults. Pb exposure (5-10 mM, 5 d) among young flies caused robust oxidative stress as evidenced by markedly elevated levels of reactive oxygen species with concomitant perturbations in the activities of antioxidant enzymes (diminished SOD and elevated thioredoxin reductase) and altered redox state. Further, Pb caused significant elevation in the activity of acetylcholinesterase and dopamine levels. In a satellite study, we assessed the modulatory effect of CSN-enriched diet (1-2%) on Pb intoxication in terms of lethality, hyperactivity, oxidative stress and neurotoxicity. CSN markedly offset Pb-induced lethality and diminished the hyperactivity response. While CSN enrichment among Pb (5 mM) treated flies caused further elevation in ROS levels and thioredoxin reductase activity, the SOD levels were restored to normalcy. Further, CSN improved the activity levels of complex I-III and restored the dopamine levels. Our data suggest that Pb-induced toxicity in the Drosophila system may be predominantly mediated through oxidative stress mechanisms and the propensity of casein-enriched diet to abrogate such responses. Hence, we propose that enrichment of diet with protein such as casein may be a useful approach to alleviate Pb associated adverse effects in children., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The risks imposed by novel insecticides, mainly bioinsecticides, are largely unknown despite their increased use and their perceived environmental safety, which is based on their natural origin. Furthermore, unlike honeybees, native pollinator species have received little attention. In the present study, the lethal and sublethal effects of the neonicotinoid imidacloprid and the bioinsecticide spinosad were assessed in the stingless bee species Meliponaquadrifasciata, an important native pollinator in the Neotropical region. The adult stingless bee workers exhibited high oral insecticide susceptibility, with LD50s of 23.54 and 12.07 ng a.i./bee for imidacloprid and spinosad, respectively. Imidacloprid also impaired worker respiration and overall group activity and flight, while spinosad significantly impaired only worker flight despite exhibiting higher oral toxicity to adult workers than imidacloprid. These findings indicate the hazardous nature not only of imidacloprid but also the bioinsecticide spinosad to adult workers of the native pollinator M. quadrifasciata. Therefore, bioinsecticides should not be exempted from risk assessment analysis due to their lethal and sublethal components., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In moths, which include many agricultural pest species, males are attracted by female-emitted sex pheromones. Although integrated pest management strategies are increasingly developed, most insect pest treatments rely on widespread use of neurotoxic chemicals, including neonicotinoid insecticides. Residual accumulation of low concentrations of these insecticides in the environment is known to be harmful to beneficial insects such as honey bees. This environmental stress probably acts as an "info-disruptor" by modifying the chemical communication system, and therefore decreases chances of reproduction in target insects that largely rely on olfactory communication. However, low doses of pollutants could on the contrary induce adaptive processes in the olfactory pathway, thus enhancing reproduction. Here we tested the effects of acute oral treatments with different low doses of the neonicotinoid clothianidin on the behavioral responses to sex pheromone in the moth Agrotis ipsilon using wind tunnel experiments. We show that low doses of clothianidin induce a biphasic effect on pheromone-guided behavior. Surprisingly, we found a hormetic-like effect, improving orientation behavior at the LD20 dose corresponding to 10 ng clothianidin. On the contrary, a negative effect, disturbing orientation behavior, was elicited by a treatment with a dose below the LD0 dose corresponding to 0.25 ng clothianidin. No clothianidin effect was observed on behavioral responses to plant odor. Our results indicate that risk assessment has to include unexpected effects of residues on the life history traits of pest insects, which could then lead to their adaptation to environmental stress.

The insect-machine interface (IMI) is a novel approach developed for man-made air vehicles, which directly controls insect flight by either neuromuscular or neural stimulation. In our previous study of IMI, we induced flight initiation and cessation reproducibly in restrained honeybees (Apis mellifera L.) via electrical stimulation of the bilateral optic lobes. To explore the neuromechanism underlying IMI, we applied electrical stimulation to seven subregions of the honeybee brain with the aid of a new method for localizing brain regions. Results showed that the success rate for initiating honeybee flight decreased in the order: α-lobe (or β-lobe), ellipsoid body, lobula, medulla and antennal lobe. Based on a comparison with other neurobiological studies in honeybees, we propose that there is a cluster of descending neurons in the honeybee brain that transmits neural excitation from stimulated brain areas to the thoracic ganglia, leading to flight behavior. This neural circuit may involve the higher-order integration center, the primary visual processing center and the suboesophageal ganglion, which is also associated with a possible learning and memory pathway. By pharmacologically manipulating the electrically stimulated honeybee brain, we have shown that octopamine, rather than dopamine, serotonin and acetylcholine, plays a part in the circuit underlying electrically elicited honeybee flight. Our study presents a new brain stimulation protocol for the honeybee-machine interface and has solved one of the questions with regard to understanding which functional divisions of the insect brain participate in flight control. It will support further studies to uncover the involved neurons inside specific brain areas and to test the hypothesized involvement of a visual learning and memory pathway in IMI flight control.

Mercury is a widespread and persistent environmental contaminant that occurs in aquatic and terrestrial habitats. Recently, songbirds that forage from primarily terrestrial sources have shown evidence of bioaccumulation of mercury, but little research has assessed the effects of mercury on their health and fitness. There are many indications that mercury negatively affects neurological functioning, bioenergetics, and behavior through a variety of mechanisms and in a wide array of avian taxa. Effective flight is crucial to avian fitness and feather molt is an energetically expensive life history trait. Therefore, we investigated whether mercury exposure influenced flight performance and molt in a common songbird, the European starling (Sturnus vulgaris). Specifically, we dosed the diet of captive starlings with methylmercury cysteine at 0.0, 0.75, or 1.5 μg/g wet weight and recorded changes in flight performance after 1 year of dietary mercury exposure. We also recorded the annual molt of wing feathers. We found that individuals dosed with mercury exhibited decreased escape takeoff flight performance compared with controls and blood mercury was also correlated with an increased rate of molt, which can reduce flight performance and thermoregulatory ability. This study reveals two novel endpoints, flight performance and molt, that may be affected by dietary mercury exposure. These findings suggest a potential impact on wild songbirds exposed to mercury levels comparable to the high dosage levels in the present study. Any decrease in flight efficiency could reduce fitness due to a direct impact on survival during predation events or by decreased efficiency in other critical activities (such as foraging or migration) that require efficient flight.

To avoid poisoning and death when toxins are ingested, the body responds with a suite of physiological detoxification mechanisms accompanied by behaviours that in mammals often include vomiting, nausea, and lethargy. Few studies have characterised whether insects exhibit characteristic 'malaise-like' behaviours in response to intoxication. Here, we used the honeybee to investigate how intoxication produced by injection or ingestion with three toxins with different pharmacological modes of action quinine, amygdalin, and lithium chloride affected behaviour. We found that toxin-induced changes in behaviour were best characterised by more time spent grooming. Bees also had difficulty performing the righting reflex and exhibited specific toxin-induced behaviours such as abdomen dragging and curling up. The expression of these behaviours also depended on whether a toxin had been injected or ingested. When toxins were ingested, they were least 10 times less concentrated in the haemolymph than in the ingested food, suggesting that their absorption through the gut is strongly regulated. Our data show that bees exhibit changes in behaviour that are characteristic of 'malaise' and suggest that physiological signalling of toxicosis is accomplished by multiple post-ingestive pathways in animals.