The role of contact chemoreception in the host location process of an egg parasitoid

Taste allows insects to detect palatable or toxic foods, identify a mate, and select appropriate oviposition sites. The gustatory system strongly contributes to the survival and reproductive success of many species, yet it is rarely studied in insect parasitoids. In order to locate and assess a host in which they will lay their eggs, female wasps actively search for chemical cues using their sensory organs present mainly on the antennae. In this paper, we studied the role of antennal taste sensilla chaetica in the perception of contact semiochemicals in Trissolcus brochymenae (Hymenoptera: Platygastridae), an egg parasitoid of the brassicaceae pest Murgantia histrionica (Heteroptera: Pentatomidae). Methanolic extracts obtained from male and female hosts elicited action potentials in taste neurons housed in antennal sensilla chaetica, indicating that these sensilla are involved in the perception of non volatile host kairomones. In behavioural assays, wasp females displayed an intense searching behaviour in open arenas treated with host extracts, thus confirming that these kairomones are soluble in polar solvents. We further investigated the extracts by Gas Chromatography-Mass Spectrometry (GC-MS) and found that they contain several compounds which are good candidates for these contact kairomones. This study contributes to better understanding contact chemoreception in egg parasitoids and identifying gustatory receptor neurons involved in the host location process.     

A female‐produced short‐range sex pheromone in the egg parasitoid Trissolcus brochymenae

The potential for short‐range sex pheromone communication by the egg parasitoid wasp Trissolcus brochymenae (Hymenoptera: Platygastridae) was investigated in closed arena bioassays. Males of this parasitoid showed more antennal drumming and more frequent mounting behaviour on 1‐ to 2‐d‐old virgin females compared with 8‐d‐old virgin females. Male copulation attempts were fewer with previously mated females than with virgin females. Males courted and made copulation attempts with 1‐ to 2‐d‐old female cadavers, but not with male cadavers or with female cadavers rinsed in organic solvents of different polarities. Male attraction to female cadavers was re‐established by treating cadavers with acetone extracts of females, but not with ether or hexane extracts. In experiments using female cadavers dissected into head, mesosoma, and gaster, and then reassembled using one unwashed body section and two body sections washed in acetone, males were attracted only to the reassembled cadavers with an unwashed mesosoma. These findings suggest that (1) courtship behaviour in males of T. brochymenae is triggered by a short‐range sex pheromone produced by females; (2) the age and the physiological condition of females (virgin/mated) influence pheromone release or production; (3) the female's mesosoma is the source of the sex pheromone; and (4) polar components of the sex pheromone play a major role in influencing male behaviour. Our results suggest that quasi‐gregarious egg parasitoids are selected for short‐range rather than long‐range sex pheromones.     

Closely linked lesions in a region of the X chromosome affect central and peripheral steps in gustatory processing in Drosophila

Summary We have analyzed a set of closely linked mutations on the X chromosome of Drosophila that lead to defects in gustatory behavior. The mutations map to a small region of the X chromosome between 10E1–4. Two distinct complementation groups, gustB and gustD, map to the ends of this region. These groups show complex complementation patterns with the mutations gustC and GT-1, which also map to this region. We describe the behavioral and electrophysiological properties of the mutants. These mutations affect peripheral receptor properties as well as more central processing steps in the gustatory pathway.     

Antennal and behavioural responses of Heortia vitessoides females to host plant volatiles of Aquilaria sinensis

This study investigated volatiles from Aquilaria sinensis (Lour.) Gilg (Thymelaeaceae) leaves that attracted Heortia vitessoides Moore (Lepidoptera: Crambidae). Volatiles from young and old A. sinensis leaves were identified and quantified by gas chromatography–mass spectrometry and gas chromatography‐electroantennogram detection. Both wind tunnel bioassays and field tests were conducted to measure the attraction of adults to synthetic blends of volatiles from leaves of different maturations. Consistent electroantennographic activity was obtained for nine and three compounds from headspace collections of young and old A. sinensis leaves, respectively. Qualitative and quantitative differences were found among two odour profiles. In wind tunnel experiments, the fresh young leaves proved to be more attractive to females than old leaves. A nine‐component mixture, including hexanal, limonene, 2‐hexanol, octanal, (Z)‐3‐hexenyl acetate, (Z)‐3‐hexen‐1‐ol, nonanal, decanal, and 2,6,10‐trimethyl‐dodecane (with a ratio of 2:16:9:4:63:100:13:10:5) from young leaves attracted moths significantly more than the three‐component mixture of nonanal, decanal, and 2,6,10‐trimethyl‐dodecane (with a ratio of 11:14:26) from old leaves. Further subtractive bioassays conducted in the wind tunnel showed that both the complete nine‐component mixture and a subtracted four‐component mixture of hexanal, (Z)‐3‐hexenyl acetate, nonanal, and decanal (with a ratio of 2:63:13:10) elicited equivalent responses in females. All components in the four‐component blend were essential for optimal attraction. In a field trial using the nine‐ and four‐component blends, more moths were captured using both blends than in traps baited with hexane only. Our study indicates that the odour blends of young leaves play an important role in H. vitessoides host plant recognition. The mechanisms behind host recognition and age‐dependent changes in leaf chemistry are discussed.     

Behavioural correlates of phenotypic plasticity in mouthpart chemoreceptor numbers in locusts

Rearing locusts in an impoverished chemosensory environment leads to fewer chemoreceptors developing on the mouthparts and antennae as adults but the behavioural relevance of these changes remains unknown. To address this question, locusts were reared for the final two larval stadia on either a single, nutritionally near-optimal synthetic food ('plain' pretreatment), or a diet comprising two nutritionally complementary foods containing two added flavours ('mix' pretreatment). Insects reared on the 'mix' diet had a mean 20% more chemosensilla on the maxillary palps than those fed on the 'plain' diet. Following an equilibration period, when all newly moulted adults could feed on two nutritionally complementary foods, insects were food deprived for 2 or 6 h, and then given a test meal of a single balanced food at one of two dilutions whilst their behaviour was recorded. 'Mix'-pretreated locusts had a shorter latency to feed and were more likely to reject the test food upon first contact if deprived for only 2 h; but if they did take a meal it lasted longer and contained fewer pauses. Using sensilla number as a covariate removed the statistical significance of pretreatment regime, indicating that sensilla number, or some close correlate of it, can largely account for the variation in behaviour. This suggests that sensilla numbers are behaviourally relevant; particularly where locusts are not greatly food deprived and faced with marginally acceptable foods.     

Tolerance to fermentation products in sugar reception: gustatory adaptation of adult butterfly proboscis for feeding on rotting foods

Adult Vanessa indica and Argyreus hyperbius frequently forage on flower nectar, but the former also utilizes tree sap and rotting fruits. Compared to flower nectar, these rotting foods are characterized by low sugar concentrations and the presence of fermentation products (ethanol and acetic acid). We suspected that gustatory responses by the receptors on the proboscis might differ in these species. Among the three sugars tested, sucrose elicited the greatest probing (behavioral) responses and was followed by fructose and glucose. A. hyperbius showed higher sugar sensitivity than V. indica in probing responsiveness. In electrophysiological responses of the proboscis sensilla, V. indica was slightly more sensitive than A. hyperbius to glucose and lower concentrations of the other sugars. The sugar reception in A. hyperbius was strongly inhibited by fermentation products, particularly acetic acid at natural concentrations. In contrast, V. indica was noticeably less susceptible to them than A. hyperbius, and its behavioral and sensory responses to sucrose were enhanced by 5-20% (w/v) ethanol. Thus, V. indica not only possesses tolerance to fermentation products but may perceive them as synergists for sugar reception. To utilize rotting foods, such tolerance might be more necessary than high sugar sensitivity.     

Sugar cell responses to lactose and sucrose in labellar and tarsal taste hairs of Musca domestica

Receptor cell responses in the largest labellar (LL) and tarsal (D) taste hairs of the housefly Musca domestica were investigated electrophysiologically using the tip-recording technique. In LL hairs, test series with lactose in concentrations of 12.5–400 mmol · l⁻¹ yielded a threshold concentration around 12 mmol · l⁻¹ and a calculated concentration eliciting half-maximal response of around 40 mmol · l⁻¹, the maximal response varying between 18 and 30 impulses/300 ms. D hairs are more sensitive towards lactose, indicated by a slightly lower threshold and a by 60% higher response to 400 mmol · l⁻¹ lactose. The high variation in the relative stimulating effectiveness of lactose and sucrose and experiments with sugar mixtures imply that these sugars bind to different receptor sites without noticeable cross affinity. A comparison of the concentration response characteristics for sucrose and lactose in LL and D hairs suggests that sucrose can combine with more than one site type, expressed in different proportions in both hair types. Results obtained with p-nitrophenyl-β-galactoside as stimulus indicate that a β-galactoside link is not sufficient for a substance to interact specifically with the lactose binding site. The exceptional lactose sensitivity of the sugar cell in M. domestica is discussed in the context of food acquirement and digestion. Accepted: 14 November 1997     

Morphology,ultrastructure and functional role of antennal sensilla in off-host aggregation by the bed bug,Cimex lectularius

After blood feeding on a host, bed bugs, Cimex lectularius, assemble in aggregation sites away from the host. Off-host aggregation is mediated by a combination of mechanical and chemical stimuli associated with bug feces. Partial antennectomies indicated removal of flagellomeres did not affect aggregation, but removal of the whole pedicel or its distal half significantly reduced (P < 0.01) aggregation, suggesting that sensilla related to off-host aggregation occur on the distal half of the pedicel. Scanning electron microscopy (SEM) revealed that serrated hairs were distributed throughout the pedicel, but newly described smooth hairs were present mainly on the distal half, and a distinct patch of grooved pegs, smooth pegs and immersed cones was present on the posterior edge of the distal half of the pedicel in adults, but not in nymphs. Numbers of different types of sensilla increased significantly during metamorphosis from first instar to adult (P < 0.05), but were similar between genders (P = 0.11) and between females from a laboratory and field strain of bugs (P = 0.19). Transmission electron microscopy (TEM) revealed that cuticular pores were present in the two types of pegs, indicating that the pegs have an olfactory function. The smooth hairs resembled gustatory sensilla previously described in Cimex hemipterus F. The existence of both olfactory and gustatory sensilla on the distal half of the pedicel suggests those sensilla may be the sensory basis of off-host aggregation behavior.     

Electrophysiological responses of receptor neurons in mosquito maxillary palp sensilla to carbon dioxide

Sensilla basiconica on the maxillary palps of female Aedes aegypti contain a receptor neuron which produces a phasic-tonic pattern of action potential response to low concentrations (150–300 ppm) of carbon dioxide (CO₂), a stimulus known to be involved with host seeking behavior. These receptor neurons respond reliably to small increments in CO₂ concentration (e.g., 50 ppm). We were particularly interested in evaluating the possibility that the sensitivity to step increases in CO₂ concentration could be modulated by alterations in the background levels of CO₂, over a range which might be encountered during host-seeking behavior. We report here that the response (impulses/s) to a single pulse of a given concentration of CO₂ appears to be independent of the background level of CO₂, unless that level is equal to or greater than the concentration of the stimulus pulse. Females of other mosquito species, including: Anopheles stephensi, Culex quinquefasciatus, Culiseta melanura, and Aedes taeniorhynchus, also possess sensilla with receptor neurons that respond with comparable sensitivity to CO₂ stimulation. However, there is much interspecific variation in both the external morphology of the maxillary palp and the distribution of sensilla along the palp. Male Ae. aegypti have morphologically similar sensilla which also contain a receptor neuron that responds to CO₂.     

Chemosensory basis of behavioural plasticity in response to deterrent plant chemicals in the larva of the Small Cabbage White butterfly Pieris rapae

Behavioural and electrophysiological responsiveness to three chemically different secondary plant substances was studied in larvae of Pieris rapae L. (Lepidoptera: Pieridae). Three groups of caterpillars were studied that during their larval development were exposed to different rearing diets: an artificial diet or one of two host-plants, cabbage, Brassica oleracea, or nasturtium, Tropaeolum majus. In dual-choice leaf disc assays, caterpillars reared on cabbage were strongly deterred by the phenolic chlorogenic acid, the flavonol glycoside naringin and the alkaloid strychnine. However, behavioural plasticity was found in caterpillars reared on nasturtium or artificial diet in that these did not discriminate against chlorogenic acid. Caterpillars reared on the artificial diet were also significantly less sensitive to naringin and strychnine in the behavioural assay. Electrophysiological studies of the maxillary sensilla styloconica revealed that the deterrent neuron in the medial sensillum, but not in the lateral sensillum, of cabbage-reared caterpillars was more sensitive than the same neuron type of caterpillars reared on nasturtium or artificial diet. We conclude that (1) the diet-induced behavioural habituation to deterrents can at least partly be explained by chemosensory desensitisation of a generalist type of maxillary deterrent neuron; (2) behavioural cross-habituation to the three structurally diverse deterrent compounds can be traced back to cross-sensitivity for these compounds in the same gustatory neuron.     

Antennal sucrose perception in the honey bee (Apis mellifera L.): behaviour and electrophysiology

Physiological mechanisms of antennal sucrose perception in the honey bee were analysed using behavioural and electrophysiological methods. Following sucrose stimulation of the tip of a freely moving antenna, the latency of proboscis extension was 320–340 ms, 80–100 ms after the first activity in muscle M17 controlling this response. When bees were allowed to actively touch a sucrose droplet with one antenna, contacts with the solution were frequent with durations of 10–20 ms and average intervals between contacts of approximately 40 ms. High sucrose concentrations led to short and frequent contacts. The proboscis response and M17 activity were largely independent of stimulus duration and temporal pattern. Taste hairs of the antennal tip displayed spike responses to sucrose concentrations down to at least 0.1%. The first 25 ms of the response were suitable for discrimination of sucrose concentrations. This time interval corresponds to the duration of naturally occurring gustatory stimuli. Sucrose responses between different hairs on the same antenna showed a high degree of variability, ranging from less than five to over 40 spikes per 0.5 s for a stimulus of 0.1% sucrose. This variability of receptor responses extends the dynamic range of sucrose perception over a large range of concentrations.     

Genetic variation in the behavioural mechanisms involved in the response of the egg parasitoid Trissolcus brochymenae to contact chemical cues left by the pest Murgantia histrionica

1. The ability of parasitoid females to perceive chemical traces left by their hosts is of utmost importance in the host location process. The behaviours involved in such ability have thus most likely been promoted by natural selection in the course of the evolutionary time. For this to happen, however, there must be significant genetic variation in natural populations on which natural selection could act.
2. Using the isofemale line method and motion analysis, we detected significant intra‐population genetic variation for several walking behaviour traits of the egg parasitoid Trissolcus brochymenae (Hymenoptera: Scelionidae) females responding to chemical traces left by its host Murgantia histrionica (Heteroptera: Pentatomidae).
3. Besides opening new avenues of research on the reproductive strategies, behaviour, and biological control potential of parasitoid wasps, these results also have implications for understanding their life‐history evolution in general.

     

Chemosensory neurons in the mouthparts of the spiny lobsters Panulirus argus and Panulirus interruptus (Crustacea: Decapoda)

We studied electrophysiological properties of single chemosensory neurons in the mouthparts of the spiny lobsters Panulirus argus and Panulirus interruptus to complement our growing understanding of the behavioral roles of mouthparts of decapod crustaceans. Food mixtures and 13 single compounds were used to characterize the response specificity, sensitivity, and time course of individual neurons in the endopods of maxilliped 2 and 3. Additional chemoreceptors were found in the mandibular palp and basis of maxilliped 1 but they were not characterized. Neurons were broadly tuned, with the five most potent single compounds being ammonium, adenosine-5′-monophosphate, taurine, glutamate, and aspartate. Cluster analysis indicated that the neurons constitute a heterogeneous population that could be placed into seven groups linked according to their most excitatory compound. These neurons in the mouthparts had concentration-dependent responses, with thresholds between 10⁻⁷ and 10⁻⁴ M and without saturation even at 10⁻³ or 10⁻² M. They also quickly adapted when exposed to their best compounds at 10⁻⁴ and 10⁻³ M. A comparison of the response properties of these neurons in the mouthparts with those of chemosensory neurons in other crustacean appendages shows that neurons in the mouthparts have relatively broad tuning biased toward detecting and resolving high concentrations. Based on these comparisons, we suggest a functional distinction among the chemosensors on the different appendages: long distance detection by the antennae, precise location and collection by the pereiopods, and detailed assessment of quality by the mouthparts.     

Chemoreception in four species of water mites (Acari: Hydrachnida): behavioural and morphological evidence

Silver nitrate staining of the solenidial and eupathidial sensilla on the palpi, tarsi and tibiae of legs I and II of Arrenurus acutus, Geayia ovata, Mideopsis reelfootensis and Albia caerulea, along with ultrastructural studies, showed that these setae have a porous shaft cuticle. These setae are hollow and contain dendrites in their lumens and more than one neuron is associated with each setae. Behavioural experiments supported the involvement of these sensilla in the perception of chemical cues from prey and sexual partners.     

Taste detection of phytoecdysteroids in larvae of Bombyx mori,Spodoptera littoralis and Ostrinia nubilalis

A number of plants produce significant amounts of phytoecdysteroids that can disrupt the hormonal levels of insects feeding upon them. Insects equipped with taste receptors sensitive to phytoecdysteroids are able to avoid such plants. How common is this strategy? By recording from the lateral and medial sensilla styloconica in two polyphagous species (Ostrinia nubilalis and Spodoptera littoralis) and in a monophagous species (Bombyx mori), we tested whether the receptors could detect three commonly occurring phytoecdysteroids 20-hydroxyecdysone (20E), ponasterone A (ponA) and ecdysone (E). In B. mori, 20E and ponA elicited dose-dependent responses with a threshold of 1 &mgr;M only in the medial sensilla. In O. nubilalis, 20E, E and ponA elicited responses at threshold of 1 &mgr;M in both sensilla. In S. littoralis, 20E elicited responses with a threshold of 10 &mgr;M in both sensilla. By means of behavioural choice tests, we show that 20E is an effective feeding deterrent for O. nubilalis and S. littoralis first instar larvae. This suggests that the perception of phytoecdysteroids is more common among phytophagous lepidoptera than previously thought, although their toxicity or antifeedancy varies between species.     

Avian colour perception predicts behavioural responses to experimental brood parasitism in chaffinches

Hosts of cuckoos have evolved defences allowing them to discriminate and reject parasite eggs. Mechanisms of discrimination are mostly visually mediated, and have been studied using approaches that do not account for what the receiver (i.e. host) actually can discriminate. Here, for the first time we apply a perceptual model of colour discrimination to study behavioural responses to natural variation in parasite egg appearance in chaffinches Fringilla coelebs. Discrimination of parasite eggs gradually increased with increasing differences in chromatic contrasts as perceived by birds between parasite and host eggs. These results confirm that colour differences of the eggs as perceived by birds are important integral parts of a matching signal used by chaffinch hosts.     

Chemoreception in four species of water mites (Acari: Hydrachnida): behavioural and morphological evidence

Silver nitrate staining of the solenidial and eupathidial sensilla on the palpi, tarsi and tibiae of legs I and II of Arrenurus acutus, Geayia ovata, Mideopsis reelfootensis and Albia caerulea, along with ultrastructural studies, showed that these setae have a porous shaft cuticle. These setae are hollow and contain dendrites in their lumens and more than one neuron is associated with each setae. Behavioural experiments supported the involvement of these sensilla in the perception of chemical cues from prey and sexual partners.     

Central gustatory projections and side-specificity of operant antennal muscle conditioning in the honeybee

Gustatory stimuli to the antennae, especially sucrose, are important for bees and are employed in learning paradigms as unconditioned stimulus. The present study identified primary antennal gustatory projections in the bee brain and determined the impact of stimulation of the antennal tip on antennal muscle activity and its plasticity. Central projections of antennal taste hairs contained axons of two morphologies projecting into the dorsal lobe, which is also the antennal motor centre. Putative mechanosensory axons arborised in a dorso-lateral area. Putative gustatory axons projected to a ventro-medial area. Bees scan gustatory and mechanical stimuli with their antennae using variable strategies but sensory input to the motor system has not been investigated in detail. Mechanical, gustatory, and electrical stimulation of the ipsilateral antennal tip were found to evoke short-latency responses in an antennal muscle, the fast flagellum flexor. Contralateral gustatory stimulation induced smaller responses with longer latency. The activity of the fast flagellum flexor was conditioned operantly by pairing high muscle activity with ipsilateral antennal sucrose stimulation. A proboscis reward was unnecessary for learning. With contralateral antennal sucrose stimulation, conditioning was unsuccessful. Thus, muscle activity induced by gustatory stimulation was important for learning success and conditioning was side-specific.     

Clonidine effects on protein and carbohydrate electrophysiological responses of labellar and tarsal sensilla in Phormia regina

The electrophysiological response of labellar and tarsal chemosensilla in the blowfly Phormia regina was studied in response to a complex stimulus naturally encountered by flies such as sheep faeces, and to beef liver, a proteinaceous feeding source. Responses were investigated both before or after injection of clonidine, an octopamine agonist previously shown to enhance sucrose ingestion, while decreasing that of proteins. As assessed by single sensillum recordings, the four different chemosensory - "salt", "sugar", "deterrent" and "water" - cells were all activated by both stimuli, regardless of sex and sensillum type, the "sugar" one being in all cases the most sensitive to beef liver before clonidine injection. Clonidine treatment affected neither labellar nor tarsal sensitivity to sucrose. Conversely, clonidine-injected flies showed a significant increase in the activity of the "deterrent" cell to beef liver, thus accounting for a decrease in protein ingestion. This study for the first time provides evidence of a key role of a clonidine-sensitive peripheral taste sensitivity in down-regulation of protein ingestion in blowflies. Correlation between peripheral sensitivity and behavioural output is discussed.