Species-Specific Induction of Plant Volatiles by Two Aphid Species in Apple: Real Time Measurement of Plant Emission and Attraction of Lacewings in the Wind Tunnel.

Upon damage by herbivores, plants release herbivory-induced plant volatiles (HIPVs). To find their prey, the pest's natural enemies need to be fine-tuned to the composition of these volatiles. Whereas standard methods can be used in the identification and quantitation of HIPVs, more recently introduced techniques such as PTR-ToF-MS provide temporal patterns of the volatile release and detect additional compounds. In this study, we compared the volatile profile of apple trees infested with two aphid species, the green apple aphid Aphis pomi, and the rosy apple aphid Dysaphis plantaginea, by CLSA-GC-MS complemented by PTR-ToF-MS. Compounds commonly released in conjunction with both species include nonanal, decanal, methyl salicylate, geranyl acetone, (Z)-3-hexenyl acetate, (Z)-3-hexenyl butanoate, (Z)-3-hexenyl 2-methyl-butanoate, (E)-ß-caryophyllene, ß-bourbonene and (Z)-3-hexenyl benzoate. In addition, benzaldehyde and (E)-ß-farnesene were exclusively associated with A. pomi, whereas linalool, (E)-4,8-dimethyl-1,3,7-nonatriene were exclusively associated with D. plantaginea. PTR-ToF-MS additionally detected acetic acid (AA) and 2-phenylethanol (PET) in the blends of both trees attacked by aphid species. In the wind tunnel, the aphid predator, Chrysoperla carnea (Stephens), responded strongly to a blend of AA and PET, much stronger than to AA or PET alone. The addition of common and species-specific HIPVs did not increase the response to the binary blend of AA and PET. In our setup, two host-associated volatiles AA + PET appeared sufficient in the attraction of C. carnea. Our results also show the importance of combining complementary methods to decipher the odor profile associated with plants under pest attack and identify behaviourally active components for predators.

A zooprophylaxis strategy using L-lactic acid (Abate) to divert host-seeking malaria vectors from human host to treated non-host animals.

BACKGROUND: Zooprophylaxis is a technique in which blood-seeking vectors are diverted to non-host animals in order to lower blood-feeding rates on human hosts. The success of this technique depends on the host preference of the vector being targeted. The objective of this study was to evaluate the effect of L-lactic acid (Abate) to divert malaria mosquito, Anopheles gambiae from feeding on human host. METHODS: A 14-month-old female goat was treated with Abate, a formulation incorporating L-lactic acid into a slow-release matrix. This formulation was applied on the fur of the goat's back and neck. The treated animal was then presented to Anopheles gambiae sensu stricto (s.s.) as a prospective host in a semi-field environment ('mosquito sphere') together with either an untreated animal or a human. The number of mosquitoes caught to each host choice offered were compared. RESULTS: Goat treated with the L-lactic acid formulation successfully attracted An. gambiae at higher rates (70.2%) than the untreated ones (29.8%). Furthermore, An. gambiae s.s. were attracted to a treated goat at an equivalent degree (47.3%) as to their preferred human host (52.7%), even when the preferred host was present in the same environment. CONCLUSIONS: The findings indicate that human host-seeking mosquitoes can be diverted into feeding on non-preferred hosts despite the close proximity of their favoured host, hence reducing chances for the transmission of blood-borne parasites.

Is Anopheles gambiae attraction to floral and human skin-based odours and their combination modulated by previous blood meal experience?

BACKGROUND: Mosquitoes use odours to find energy resources, blood hosts and oviposition sites. While these odour sources are normally spatio-temporally segregated in a mosquito's life history, here this study explored to what extent a combination of flower- and human-mimicking synthetic volatiles would attract the malaria vector Anopheles gambiae sensu stricto (s.s.) METHODS: In the laboratory and in large (80 m2) outdoor cages in Tanzania, nulliparous and parous A. gambiae s.s. were offered choices between a blend of human skin volatiles (Skin Lure), a blend of floral volatiles (Vectrax), or a combination thereof. The blends consisted of odours that induce distinct, non-overlapping activation patterns in the olfactory circuitry, in sensory neurons expressing olfactory receptors (ORs) and ionotropic receptors (IRs), respectively. Catches were compared between treatments. RESULTS: In the laboratory nulliparous and parous mosquitoes preferred skin odours and combinations thereof over floral odours. However, in semi-field settings nulliparous were significantly more caught with floral odours, whereas no differences were observed for parous females. Combining floral and human volatiles did not augment attractiveness. CONCLUSIONS: Nulliparous and parous A. gambiae s.s. are attracted to combinations of odours derived from spatio-temporally segregated resources in mosquito life-history (floral and human volatiles). This is favourable as mosquito populations are comprised of individuals whose nutritional and developmental state steer them to diverging odours sources, baits that attract irrespective of mosquito status could enhance overall effectiveness and use in monitoring and control. However, combinations of floral and skin odours did not augment attraction in semi-field settings, in spite of the fact that these blends activate distinct sets of sensory neurons. Instead, mosquito preference appeared to be modulated by blood meal experience from floral to a more generic attraction to odour blends. Results are discussed both from an odour coding, as well as from an application perspective.

Translating olfactomes into attractants: shared volatiles provide attractive bridges for polyphagy in fruit flies.

Tephritid flies are serious fruit pests. Despite clear niche differences, many species show considerable overlap in fruit preferences, of which we here analysed the olfactory correlate. Using the volatiles of four unrelated fruit species, antennal responses were quantified to construct a fruit-odour response database for four tephritid species. Although responses were distinct with a significant niche-correlated bias, the analyses show that the probability of detection of a volatile strongly increased with its sharedness across fruits. This also held for the unrelated fruit fly Drosophila melanogaster (DoOR repository-based analyses). We conjectured that shared volatiles signify 'host' to the fly 'nose' and induce attraction. Indeed, blends of volatiles shared by fruit and detected by all four species were very attractive for tephritid species, more than fruits. Quantitative whole antennal recordings en lieu of, or complementing bottom-up molecular neurogenetic approaches, enables comparative olfactomics in non-model species, and facilitate interpretation of olfaction in evolutionary, ecological, and applied contexts.

Genetic mapping of male pheromone response in the European corn borer identifies candidate genes regulating neurogenesis.

The sexual pheromone communication system of moths is a model system for studies of the evolution of reproductive isolation. Females emit a blend of volatile components that males detect at a distance. Species differences in female pheromone composition and male response directly reinforce reproductive isolation in nature, because even slight variations in the species-specific pheromone blend are usually rejected by the male. The mechanisms by which a new pheromone signal-response system could evolve are enigmatic, because any deviation from the optimally attractive blend should be selected against. Here we investigate the genetic mechanisms enabling a switch in male response. We used a quantitative trait locus-mapping approach to identify the genetic basis of male response in the two pheromone races of the European corn borer, Ostrinia nubilalis Male response to a 99:1 vs. a 3:97 ratio of the E and Z isomers of the female pheromone is governed by a single, sex-linked locus. We found that the chromosomal region most tightly linked to this locus contains genes involved in neurogenesis but, in accordance with an earlier study, does not contain the odorant receptors expressed in the male antenna that detect the pheromone. This finding implies that differences in the development of neuronal pathways conveying information from the antenna, not differences in pheromone detection by the odorant receptors, are primarily responsible for the behavioral response differences among the males in this system. Comparison with other moth species reveals a previously unexplored mechanism by which male pheromone response can change in evolution.

The Genetic Basis of Pheromone Evolution in Moths.

Moth sexual pheromones are widely studied as a fine-tuned system of intraspecific sexual communication that reinforces interspecific reproductive isolation. However, their evolution poses a dilemma: How can the female pheromone and male preference simultaneously change to create a new pattern of species-specific attraction? Solving this puzzle requires us to identify the genes underlying intraspecific variation in signals and responses and to understand the evolutionary mechanisms responsible for their interspecific divergence. Candidate gene approaches and functional analyses have yielded insights into large families of biosynthetic enzymes and pheromone receptors, although the factors controlling their expression remain largely unexplored. Intra- and interspecific crosses have provided tantalizing evidence of regulatory genes, although, to date, mapping resolution has been insufficient to identify them. Recent advances in high-throughput genome and transcriptome sequencing, together with established techniques, have great potential to help scientists identify the specific genetic changes underlying divergence and resolve the mystery of how moth sexual communication systems evolve.

Do Fruit Ripening Volatiles Enable Resource Specialism in Polyphagous Fruit Flies?

Frugivorous tephritid fruit flies have lineages with high levels of host generalism. These insects use olfaction to locate fruits, but how they are able to recognize the odors of so many different host species is poorly understood. We used a series of behavioral experiments to investigate the role of fruit ripening volatiles as host cues in the Queensland fruit fly, Bactrocera tryoni (Froggatt), a polyphagous pest in Australia. Odors of mature guava (Psidium guajava) attracted female and male flies more strongly than three other ripening stages and guava pulp. We analyzed volatiles from guava odor and selected eleven compounds, all of which elicited an electrophysiological response in the antenna of female flies. Three of these, ethyl acetate, ethyl butyrate, and ethyl propionate, were released at the highest rates from the most attractive ripening stage. In behavioral trials, these three esters were not attractive individually, whereas a combination was necessary and sufficient in attracting female flies. The three-component blend was as attractive as the entire 11-component blend, which without these key volatiles was not attractive. Moreover, injecting low ranking hosts (squash and cucumber) with the three volatiles increased attraction in ovipositing female flies. These fruit flies are classed as generalists, but like many polyphagous insects they could be regarded as resource specialists, preferring specific plant reproductive stages with predictable odor cues. Exploring olfaction from this perspective could improve our understanding of host choice in polyphagous insects, and the selection of volatiles to be used as attractants in insect pest management.

Early quality assessment lessens pheromone specificity in a moth.

Pheromone orientation in moths is an exemplar of olfactory acuity. To avoid heterospecific mating, males respond to female-produced blends with high specificity and temporal resolution. A finely tuned sensory to projection neuron network secures specificity, and this network is thought to assess pheromone quality continually during orientation. We tested whether male moths do indeed evaluate each pheromone encounter and surprisingly found that male European corn borer moths instead generalize across successive encounters. Although initially highly ratio specific, once "locked on" to the pheromone plume the acceptable ratio can vary widely, and even unattractive blends can become attractive. We further found that this "mental shortcut" may be a consequence of the fact that sensory neurons exposed to frequent encounters do not reliably encode blend ratios. Neurons tuned to either of the two pheromone components adapt differentially in plumes containing the preferred blend ratio (97:3) and cause the olfactory sensory signal to "evolve," even in narrowly tuned pheromonal circuits. However, apparently the brain interprets these shifting signals as invariant "gestalts." Generalization in pheromone perception may mitigate stabilizing selection and allow introgression between sympatric strains, such as in the European corn borer, that otherwise appear isolated by pheromonal differences. Generalization may also be important in responses to general odorants, as circuits underlying these display vast sensitivity differences, complex interactions, and temporal intricacies.

A herbivore-induced plant volatile interferes with host plant and mate location in moths through suppression of olfactory signalling pathways.

BACKGROUND: Plants under herbivore attack release volatiles that attract natural enemies, and herbivores in turn avoid such plants. Whilst herbivore-induced plant volatile blends appeared to reduce the attractiveness of host plants to herbivores, the volatiles that are key in this process and particularly the way in which deterrence is coded in the olfactory system are largely unknown. Here we demonstrate that herbivore-induced cotton volatiles suppress orientation of the moth Spodoptera littoralis to host plants and mates. RESULTS: We found that (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), an induced volatile, is key in herbivore deterrence: DMNT suppressed plant odour- and pheromone-induced behaviours. We then dissected the neurophysiological basis of this interaction. DMNT-responding glomeruli were also activated by other plant compounds, suggesting that S. littoralis possesses no segregated olfactory circuit dedicated exclusively to DMNT. Instead, DMNT suppressed responses to the main pheromone component, (Z)-9-(E)-11-tetradecenyl acetate, and primarily to (Z)-3-hexenyl acetate, a host plant attractant. CONCLUSION: Our study shows that olfactory sensory inhibition, which has previously been reported without reference to an animal's ecology, can be at the core of coding of ecologically relevant odours. As DMNT attracts natural enemies and deters herbivores, it may be useful in the development or enhancement of push-pull strategies for sustainable agriculture.

Loss of Drosophila pheromone reverses its role in sexual communication in Drosophila suzukii.

The Drosophila pheromone cis-11-octadecenyl acetate (cVA) is used as pheromone throughout the melanogaster group and fulfils a primary role in sexual and social behaviours. Here, we found that Drosophila suzukii, an invasive pest that oviposits in undamaged ripe fruit, does not produce cVA. In fact, its production site, the ejaculatory bulb, is atrophied. Despite loss of cVA production, its receptor, Or67d, and cognate sensillum, T1, which are essential in cVA-mediated behaviours, were fully functional. However, T1 expression was dramatically reduced in D. suzukii, and the corresponding antennal lobe glomerulus, DA1, minute. Behavioural responses to cVA depend on the input balance of Or67d neurons (driving cVA-mediated behaviours) and Or65a neurons (inhibiting cVA-mediated behaviours). Accordingly, the shifted input balance in D. suzukii has reversed cVA's role in sexual behaviour: perfuming D. suzukii males with Drosophila melanogaster equivalents of cVA strongly reduced mating rates. cVA has thus evolved from a generic sex pheromone to a heterospecific signal that disrupts mating in D. suzukii, a saltational shift, mediated through offsetting the input balance that is highly conserved in congeneric species. This study underlines that dramatic changes in a species' sensory preference can result from rather 'simple' numerical shifts in underlying neural circuits.

Electrophysiologically-Active Maize Volatiles Attract Gravid Female European Corn Borer, Ostrinia nubilalis.

The European corn borer (ECB) is an important pest of maize in the northern hemisphere, but no reliable techniques exist for monitoring females during their reproductive period. In this study, we aimed to identify host-plant volatiles used by gravid Z-strain females in search for oviposition sites. Headspace of maize plants, to which gravid females orientated in a wind tunnel, was collected, and physiologically-active components were identified by using gas chromatography (GC) coupled with electroantennographic detection followed by GC-mass spectrometry. The antennae of female moths consistently responded to two maize volatiles, nonanal and decanal. Although these compounds are individually not characteristic for maize, a synthetic mix in a ratio found in maize headspace, 1:2.4 at 1 µg µl(-1) induced source contact and landing responses similar to maize plants in the wind tunnel. However, fewer females took flight in response to the mix, and those that took flight did so with an increased latency. To our knowledge, this is the first blend of host-plant volatiles that has been found to be physiologically active and to be able to induce attraction of gravid ECB females under laboratory conditions. Future tests will evaluate the attractiveness of the blend to the E-strain of ECB, the attractiveness of the blend in the field, and its potential in monitoring ECB populations.

Identification of host blends that attract the African invasive fruit fly, Bactrocera invadens.

Bactrocera invadens, an invasive fruit fly species in the Afro-tropical region belonging to the Bactrocera dorsalis complex, causes considerable damage to fruit production and productivity. We sought to find attractants from hosts of B. invadens that could serve as baits in traps for monitoring and management of this pest. The attractiveness of volatiles from four different fruit species (mango, guava, banana and orange) at two stages of ripeness (ripe or unripe) was tested in an olfactometer assay. All fruits were attractive against a clean air control. Using hexane extracts of volatile collections of fruits, we demonstrated that male flies preferred the volatiles of ripe guava and orange over unripe fruit extracts. There was a slight difference in preference between females and males; females preferred orange to guava and mango, whereas males preferred mango and guava to orange. Gas chromatography/electroantennographic detection (GC/EAD) and GC/mass spectrometry (GC/MS) were used to identify compounds to which B. invadens antennae were sensitive. GC/EAD recordings from distal and medio-central parts of the fly antenna showed responses to a number of compounds from each fruit species, with esters dominating the responses. Synthetic blends were made for each fruit species using the shared antennally active compounds in ratios found in the extracts. In the olfactometer, B. invadens was most attracted to the banana and orange blends, followed by the mango and guava blends. The synthetic banana blend was as attractive as the volatile collection of banana, although both were less attractive than the fruit. The results demonstrate that composing attractive blends from GC/EAD-active constituents shared by host fruits can be effective for formulating attractive synthetic host mimics for generalist fruit fly species, such as B. invadens.

The maxillary palps of Tephritidae are selectively tuned to food volatiles and diverge with ecology.

The maxillary palp is an auxiliary olfactory organ in insects, which, different from the antennae, is equipped with only a few olfactory sensory neuron (OSN) types. We postulated that these derived mouthpart structures, positioned at the base of the proboscis, may be particularly important in mediating feeding behaviors. As feeding is spatio-temporally segregated from oviposition in most Tephritidae, this taxonomic group appears quite suitable to parse out sensory breadth and potential functional divergence of palps and antennae. Scanning electron microscopy and anterograde staining underlined the limited palpal olfactory circuit in Tephritidae: only three morphological subtypes of basiconic sensilla were found, each with two neurons, and project to a total of six antennal lobe glomeruli in Bactrocera dorsalis. Accordingly, the palps detected only few volatiles from the headspace of food (fermentation and protein lures) and fruit (guava and mango) compared to the antennae (17 over 77, using gas-chromatography coupled electrophysiology). Interestingly, functionally the antennae were more tuned to fruit volatiles, detecting eight times more fruit than food volatiles (63 over 8), whereas the number of fruit and food volatile detection was more comparable in the palps (14 over 8). As tephritids diverge in oviposition preferences, but converge on food substrates, we postulated that the receptive ranges of palpal circuits would be more conserved compared to the antennae. However, palpal responses of three tephritid species that differed in phylogenetic relatedness and ecologically niche, diverged across ecological rather than phylogenetic rifts. Two species with strongly overlapping ecology, B. dorsalis and Ceratitis capitata, showed inseparable response profiles, whereas the cucurbit specialist Zeugodacus cucurbitae strongly diverged. As Z. cucurbitae is phylogenetically placed between B. dorsalis and C. capitata, the results indicate that ecology overrides phylogeny in the evolution of palpal tuning, in spite of being predisposed to detecting food volatiles.

The push-pull intercrop Desmodium does not repel, but intercepts and kills pests.

Over two decades ago, an intercropping strategy was developed that received critical acclaim for synergizing food security with ecosystem resilience in smallholder farming. The push-pull strategy reportedly suppresses lepidopteran pests in maize through a combination of a repellent intercrop (push), commonly Desmodium spp., and an attractive, border crop (pull). Key in the system is the intercrop's constitutive release of volatile terpenoids that repel herbivores. However, the earlier described volatile terpenoids were not detectable in the headspace of Desmodium, and only minimally upon herbivory. This was independent of soil type, microbiome composition, and whether collections were made in the laboratory or in the field. Furthermore, in oviposition choice tests in a wind tunnel, maize with or without an odor background of Desmodium was equally attractive for the invasive pest Spodoptera frugiperda. In search of an alternative mechanism, we found that neonate larvae strongly preferred Desmodium over maize. However, their development stagnated and no larva survived. In addition, older larvae were frequently seen impaled and immobilized by the dense network of silica-fortified, non-glandular trichomes. Thus, our data suggest that Desmodium may act through intercepting and decimating dispersing larval offspring rather than adult deterrence. As a hallmark of sustainable pest control, maize-Desmodium push-pull intercropping has inspired countless efforts to emulate stimulo-deterrent diversion in other cropping systems. However, detailed knowledge of the actual mechanisms is required to rationally improve the strategy, and translate the concept to other cropping systems.

Mango headspace volatiles trigger differential responses of the mango fruit fly Ceratitis cosyra and its parasitoids.

Before the introduction of Bactrocera dorsalis (Hendel) to sub-Saharan Africa, Ceratitis cosyra (Walker) was economically the most important pest in mango farming. Its native natural enemy, the solitary parasitoid Psyttalia cosyrae (Wilkinson), played a crucial role in C. cosyra bio-control, later complemented by the exotic parasitoids Diachasmimorpha longicaudata (Ashmead) and Fopius arisanus (Sonan) among Integrated Pest Management (IPM) systems. To understand the in situ mango-C. cosyra-parasitoid tritrophic interaction, we assessed the responses of the fruit fly and the three parasitoids to headspace volatiles from various mango conditions. These conditions included non-infested mature unripe mangoes, C. cosyra-infested mangoes, 7th- and 9th-day post-infestation mangoes, non-infested ripe mangoes of three varieties (Kent, Apple, and Haden), and clean air (blank). We also compared the fruit fly's performance in the mango varieties and identified the chemical profiles of mango headspace volatiles. Ceratitis cosyra was attracted to both infested and non-infested mangoes (66-84 % of responsive C. cosyra) and showed superior performance in Kent mango (72.1 % of the 287 puparia recovered) compared to Apple and Haden varieties. Fopius arisanus displayed a stronger attraction to the volatiles of C. cosyra-infested mangoes (68-70 %), while P. cosyrae and D. longicaudata were significantly attracted to the 9th-day post-infestation mangoes (68-78 %) compared to non-infested mango volatiles. Gas chromatography-mass spectroscopy showed substantial quantitative and qualitative differences in volatile profiles among mango treatments. Esters predominated in non-infested ripe, 7th- and 9th-day post-infestation mangoes, while monoterpenes and sesquiterpenes were most dominant in the other treatments. The in situ experiments underscored varying preferences of the species for mango headspace volatiles and their subsequent treatments. These results provide valuable insights for further exploration, specifically in identifying the key volatiles responsible for species responses, to facilitate the development of applicable selective semiochemicals for managing species of African fruit fly.

A carboxylesterase, Esterase-6, modulates sensory physiological and behavioral response dynamics to pheromone in Drosophila.

BACKGROUND: Insects respond to the spatial and temporal dynamics of a pheromone plume, which implies not only a strong response to 'odor on', but also to 'odor off'. This requires mechanisms geared toward a fast signal termination. Several mechanisms may contribute to signal termination, among which odorant-degrading enzymes. These enzymes putatively play a role in signal dynamics by a rapid inactivation of odorants in the vicinity of the sensory receptors, although direct in vivo experimental evidences are lacking. Here we verified the role of an extracellular carboxylesterase, esterase-6 (Est-6), in the sensory physiological and behavioral dynamics of Drosophila melanogaster response to its pheromone, cis-vaccenyl acetate (cVA). Est-6 was previously linked to post-mating effects in the reproductive system of females. As Est-6 is also known to hydrolyze cVA in vitro and is expressed in the main olfactory organ, the antenna, we tested here its role in olfaction as a putative odorant-degrading enzyme. RESULTS: We first confirm that Est-6 is highly expressed in olfactory sensilla, including cVA-sensitive sensilla, and we show that expression is likely associated with non-neuronal cells. Our electrophysiological approaches show that the dynamics of olfactory receptor neuron (ORN) responses is strongly influenced by Est-6, as in Est-6° null mutants (lacking the Est-6 gene) cVA-sensitive ORN showed increased firing rate and prolonged activity in response to cVA. Est-6° mutant males had a lower threshold of behavioral response to cVA, as revealed by the analysis of two cVA-induced behaviors. In particular, mutant males exhibited a strong decrease of male-male courtship, in association with a delay in courtship initiation. CONCLUSIONS: Our study presents evidence that Est-6 plays a role in the physiological and behavioral dynamics of sex pheromone response in Drosophila males and supports a role of Est-6 as an odorant-degrading enzyme (ODE) in male antennae. Our results also expand the role of Est-6 in Drosophila biology, from reproduction to olfaction, and highlight the role of ODEs in insect olfaction.

What reaches the antenna? How to calibrate odor flux and ligand-receptor affinities.

Physiological studies on olfaction frequently ignore the airborne quantities of stimuli reaching the sensory organ. We used a gas chromatography-calibrated photoionization detector to estimate quantities released from standard Pasteur pipette stimulus cartridges during repeated puffing of 27 compounds and verified how lack of quantification could obscure olfactory sensory neuron (OSN) affinities. Chemical structure of the stimulus, solvent, dose, storage condition, puff interval, and puff number all influenced airborne quantities. A model including boiling point and lipophilicity, but excluding vapor pressure, predicted airborne quantities from stimuli in paraffin oil on filter paper. We recorded OSN responses of Drosophila melanogaster, Ips typographus, and Culex quinquefasciatus, to known quantities of airborne stimuli. These demonstrate that inferred OSN tuning width, ligand affinity, and classification can be confounded and require stimulus quantification. Additionally, proper dose-response analysis shows that Drosophila AB3A OSNs are not promiscuous, but highly specific for ethyl hexanoate, with other earlier proposed ligands 10- to 10 000-fold less potent. Finally, we reanalyzed published Drosophila OSN data (DoOR) and demonstrate substantial shifts in affinities after compensation for quantity and puff number. We conclude that consistent experimental protocols are necessary for correct OSN classification and present some simple rules that make calibration, even retroactively, readily possible.

Moment-to-moment flight manoeuvres of the female yellow fever mosquito (Aedes aegypti L.) in response to plumes of carbon dioxide and human skin odour.

Odours are crucial cues enabling female mosquitoes to orient to prospective hosts. However, their in-flight manoeuvres to host odours are virtually unknown. Here we analyzed in 3-D the video records of female Aedes aegypti mosquitoes flying in a wind tunnel in response to host odour plumes that differed in spatial structure and composition. Following a brief (~0.03 s) encounter with CO(2), mosquitoes surged upwind and, in the absence of further encounters, counterturned without displacing upwind. These patterns resemble moth responses to encounter and loss of a filament of pheromone. Moreover, CO(2) encounters induced a highly regular pattern of counterturning across the windline in the horizontal (crosswind) and vertical planes, causing the mosquito to transect repeatedly the area where CO(2) was previously detected. However, despite the rapid changes across all three axes following an encounter with CO(2), the angular velocities remained remarkably constant. This suggests that during these CO(2)-induced surges mosquitoes stabilize flight through sensors, such as the halteres and Johnston organs, sensitive to Coriolis forces. In contrast to the instantaneous responses of the mosquito CO(2), a brief encounter with a filament of human skin odour did not induce a consistent change in mosquito flight. These differential responses were reflected in further experiments with broad plumes. A broad homogeneous plume of skin odour induced rapid upwind flight and source finding, whereas a broad filamentous plume of skin odour lowered activation rates, kinetic responses and source finding compared with homogeneous plumes. Apparently, yellow fever mosquitoes need longer continuous exposure to complex skin-odour blends to induce activation and source finding.

bric à brac controls sex pheromone choice by male European corn borer moths.

The sex pheromone system of ~160,000 moth species acts as a powerful form of assortative mating whereby females attract conspecific males with a species-specific blend of volatile compounds. Understanding how female pheromone production and male preference coevolve to produce this diversity requires knowledge of the genes underlying change in both traits. In the European corn borer moth, pheromone blend variation is controlled by two alleles of an autosomal fatty-acyl reductase gene expressed in the female pheromone gland (pgFAR). Here we show that asymmetric male preference is controlled by cis-acting variation in a sex-linked transcription factor expressed in the developing male antenna, bric à brac (bab). A genome-wide association study of preference using pheromone-trapped males implicates variation in the 293 kb bab intron 1, rather than the coding sequence. Linkage disequilibrium between bab intron 1 and pgFAR further validates bab as the preference locus, and demonstrates that the two genes interact to contribute to assortative mating. Thus, lack of physical linkage is not a constraint for coevolutionary divergence of female pheromone production and male behavioral response genes, in contrast to what is often predicted by evolutionary theory.

Dispersal and competitive release affect the management of native and invasive tephritid fruit flies in large and smallholder farms in Ethiopia.

African horticulture is seriously affected by fruit flies, both native and invasive. Novel sustainable control methods need testing against the backdrop of smallholder-dominated farming of Africa. We evaluated the potential of male-specific attractants (parapheromones) laced with insecticide to suppress the alien invasive Bactrocera dorsalis and native Ceratitis capitata. In large-scale guava, methyl-eugenol (ME)-bait stations combined with toxic protein baits suppressed B. dorsalis within 8 months but resulted in a resurgence of the displaced Ceratitis capitata. In smallholder farms, intervention using SPLAT-ME laced with spinosad was surprisingly unsuccessful. Subsequent mark-release-recapture experiments showed high dispersal rates of flies, covering many times a typical farm size, leading to a continuous influx of flies from surrounding areas. Several other factors important for intervention were evaluated. SPLAT-MAT-ME dollops remained attractive for over two weeks, although gradually becoming less attractive than fresh baits. Further, competitive displacement was observed: C. capitata selectively emerged from fruits in which B. dorsalis infestation was low. Finally, we evaluated whether ME could be combined with C. capitata male attractants [trimedlure (TML) and terpinyl acetate (TA)] without affecting attraction. Combining male lures did not affect catches directly, although at very high populations of B. dorsalis attracted to ME interfered with C. capitata trap entry. Although ME-based methods can effectively suppress B. dorsalis, they were not effective at single smallholder scale due to the high dispersive propensity of tephritids. Further, competitive release implies the need for a combination of lures and methods. These observations are important for developing control schemes tailored for African smallholder settings.