Host odours enhance the responses of adult Rhyzopertha dominica to male-produced aggregation pheromone

The behavioural responses of adult male and female Rhyzopertha dominica (Coleoptera, Bostrichidae) to blends of host volatiles and male-produced aggregation pheromone were observed in a four-arm airflow olfactometer. The odour sources used were five pheromone-releasing males each on a single maize grain (lower maize-volatiles blend), five pheromone-releasing males on 500 g of maize (higher maize-volatiles blend) or the host volatiles emanating from 500g of maize (maize volatiles alone). Multiple-choice tests, in which volatiles from all three odour sources were presented in the exposure chamber at the same time, were used to study odour preferences of the males and females. Both sexes showed strongest attraction to the higher maize volatile blend but there were significant sex differences in response to the odour sources. Males spent significantly more time than females in the zone with only maize volatiles, and females spent significantly more time in the higher maize-volatiles zone. However, when odour sources were offered singly, females gave numerically greater responses than males to all sources although this difference was statistically significant for only the lower maize-volatiles blend. As males are more attracted than females to host odours alone it is suggested that they may be more highly adapted to seek out new hosts while females are more inclined to locate a food source by following the pheromone signals produced by males. However, both sexes responded most strongly to the odour source comprising aggregation pheromone with the higher proportion of maize volatiles.     

Olfactometer Responses of Plum Curculio <Emphasis Type="Italic">Conotrachelus nenuphar</Emphasis> (Herbst) (Coleoptera: Curculionidae) to Host Plant Volatiles,Synthetic Grandisoic Acid,and Live Conspecifics

The plum curculio Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae) is a major pest of pome and stone fruit, but will also attack other fruits. Males produce the aggregation pheromone grandisoic acid; emitting only the (+)-enantiomer which is attractive to both sexes of the univoltine and multivoltine strains, while the synthetic racemic mixture contains optical isomers with equal amounts of (+)- and (?)-enantiomers. Synergy between odours can increase trap captures and improve monitoring techniques, therefore tests were performed in a dual-choice olfactometer with odours attractive to plum curculios according to literature to determine 1) under what physiological conditions (mating status, age, starvation period) these odours are attractive or repulsive, 2) if the (+)-enantiomer or the odour of live males synergizes with host plant volatiles, and 3) if there is a difference in response between plum curculio strains. Females were exposed to: benzaldehyde; trans-2-hexenal; apples; extracts of: plums, apples, blueberries; grandisoic acid; and live males. Plum essence was found to be the most attractive host-plant odour for both immature and mature virgin females, and immature whole apples were attractive to starved females, while trans-2-hexenal, McIntosh apple essence, benzaldehyde along with the combination of benzaldehyde and plume essence was found to be repulsive. Starvation, age, and mated status all influence response to odours. No synergistic or additive affects were observed between any of the odour combinations tested, including the combination of both the natural and synthetic pheromone and plum essence or apples.     

A six-arm olfactometer permitting simultaneous observation of insect attraction and odour trapping

Abstract.  Behavioural assays to study insect attraction to specific odours are tedious, time consuming and often require large numbers of replications. Olfactometer and flight tunnel tests can usually only be conducted with one or two odour sources at a time. Moreover, chemical information on the odour sources has to be obtained in separate analytical studies. An olfactometer was developed in which six odours can be tested simultaneously for their relative attractiveness while during the assays, part of each test odour can be trapped for further analyses. The effectiveness of this six-arm olfactometer was tested by observing the responses of the solitary endoparasitoid Cotesia marginiventris (Cresson) to host-induced odours from young maize plants. For statistical analyses, we used log-linear models were adapted to account for overdispersion and possible positional biases. Female wasps responded extremely well in tests where they were offered a single odour source, as well as in tests with multiple choices. The responses of wasps released in groups were the same as those released individually and it was found that females did not attract or repel each other, but males preferred arms in which females had been released. Dose–response tests with varying numbers of plants or host larvae on plants revealed that the wasps responded in a dose-related manner, thus showing that the system is well suited to measure relative preference. The clear choices of the insects amongst six possibilities provided substantial statistical power. Gas chromatographic analyses of sampled air revealed clean and effective odour trapping, which largely facilitates the comparison of results from behavioural assays with the actual blends of volatiles that were emitted by the various odour sources. Advantages and disadvantages compared to other methods are discussed.     

Cereal aphid responses to sex pheromones and host-plant odours in the laboratory

Abstract. The movement of Sitobion fragariae (Wlk.) (Homoptera: Aphididae) males towards odours from conspecific sexual females (oviparae) and the aphid sex pheromone component (4a S ,7 S ,7a R )-nepetalactone were demonstrated in a linear-track olfactometer. Males of the closely related Sitobion avenae (Fab.) were attracted to odours from conspecific oviparae and also oviparae of S. fragariae. These results were consistent with the recent identification of this nepetalactone isomer as the major component of the sex pheromone in these species. Males of S.fragariae were not significantly attracted to oviparae of S.avenae. indicating some qualitative or quantitative difference in the pheromones of the two species. Males did not respond to the related (1R,4aS,7S,7aR)-nepetalactol or to host-plant odours. Gynoparae of these species moved towards the nepetalactone and also to host-plant odours. A combination of both stimuli was more attractive than plant odour alone. Gynoparae of S.fragariae also responded significantly to the combined treatment when this was tested against the nepetalactone.     

Specificity of sex pheromones, the role of host plant odour in the olfactory attraction of males, and mate recognition in the aphid Cryptomyzus

Abstract. Choice tests in an olfactometer showed that males of the aphid Cryptomyzus galeopsidis spent more time in the odour of conspecific females than of a sibling species or a more distantly related Cryptomyzus species, which all co-occur on red currant. Analysis of the pheromone-emissions of females showed the presence of a nepetalactol and a nepetalactone, two known aphid sex pheromone components. However, no interspecific difference in the ratio of the components ( c. 30:1) was found, and this ratio did not change as the females aged. Males were only slightly attracted by a synthetic 30:1 mixture of these two components, and in a choice test preferreď the odour of conspecific females. The sex pheromone of Cryptomyzus spp. must thus consist of more than two components, the first time this has been reported for aphids. Host plant odour did not attract males or enhance the activity of the pheromone.     

Different bioassays for investigating orientation responses of the banana weevil, Cosmopolites sordidus, show additive effects of host plant volatiles and a synthetic male-produced aggregation pheromone

Abstract Three different bioassay methods to investigate the orientation behaviour of the banana weevil, Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae), to host plant volatiles and a synthetic pheromone (cosmolure+) were compared. A locomotion compensator was used to separately record walking tracks in response to three odour sources. The data show that C. sordidus uses odour‐conditioned anemotaxis in its orientation to the odour sources tested. Of the two olfactometers tested, a dual port olfactometer using a continuous airflow showed stronger discrimination by C. sordidus to the different odours compared with a double pitfall olfactometer. The results of all three bioassays indicate that C. sordidus responds in an additive way to the combination of fermentation plant volatiles and the synthetic pheromone.     

Olfactory cues in discrimination among individuals in dominance hierarchies in the cockroach, Nauphoeta cinerea

ABSTRACT. The purpose of this study was to explore the olfactory recognition mechanisms used by individual subordinate male cockroaches, Nauphoeta cinerea (Olivier), in a dominance hierarchy. Results of two independent sets of experiments, one using an olfactometer and the other using an arena, suggest that olfactory cues are important to males. Subordinate males did not avoid dominant male odour in an olfactometer but could discriminate between the odours of familiar and unfamiliar individual dominant males, preferring the odour of the dominant male with which they had previous social contact. Since test subordinates did not avoid dominant male odours in the olfactometer, it is possible that the distance an individual remains from a dominant is important. To test this, individual dominant and subordinate males were tethered in the centre of an arena. Subordinate males were introduced into the arena and allowed to approach the tethered male. They approached tethered dominants and subordinates differently, and tended to keep away from the dominants. Tethered males unfamiliar to the test animals were avoided as well.     

The design of a phonoactograph suitable for study on the influence of pheromones on insect mobility

An instrument for recording changes in insect mobility under the influence of pheromone was described. The instrument was named "phonoactograph" because the intensity of rustle made by insects was utilized as a correlate of their movement. The Colorado beetle males exposed to the action of the female odour increased their mobility significantly.     

Preliminary experiments on the olfactory responses of Pterostichus melanarius Illiger (Coleoptera: Carabidae) to intact plants

1 In two successive years, the olfactory responses of Pterostichus melanarius Illiger (Coleoptera: Carabidae) to intact cabbage and white clover plants were studied, using a four‐arm olfactometer. 2 The first set of experiments tested the response to the odours of cabbage, white clover and the two plants grown together against potting compost; in the second and third sets of experiments, white clover and cabbage were tested, respectively. 3 The results indicate that female P. melanarius individuals are arrested by the combined odours of cabbage and white clover, spending significantly more time in the cabbage with white clover odour field than in either the cabbage, white clover or control odour fields. 4 Similar results were not found for males. 5 Cabbage plants alone did not elicit significant responses in either sex. 6 White clover alone did not elicit significant responses by females and the responses of males were not uniform in the two successive years.     

Divergent responses of two cereal aphids to previous infestation of their host plant

The role of pollen odour in resource location by the pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), a pollen-feeding insect regarded as a pest of oilseed rape, Brassica napus L., (Brassicaceae) crops, was investigated in a linear track olfactometer. Both male and female beetles were attracted to the odour of whole oilseed rape flowers, indicating that these insects can locate their host plants using floral odours as cues. The attractive odour of flowers was found to emanate from all floral parts tested: the petals/sepals, the anthers, and from pollen itself. Therefore, at least part of the attractive odour of oilseed rape flowers emanates from pollen. Beetles were more attracted to floral samples containing anthers than those without anthers when these odours were directly compared in a choice-test, and this indicates that there were detectable differences between them. Anthers and pollen may therefore release distinctive odours that are quantitatively and/or qualitatively different from the odour of the rest of the flower. These experiments support the hypothesis that pollen-seeking insects use pollen odour cues to locate this food source.     

Calcium responses to pheromones and plant odours in the antennal lobe of the male and female moth Heliothis virescens

In male moths, the primary olfactory integration centre, the antennal lobe, consists of two systems. The macroglomerular complex processes pheromone information, while the ordinary glomeruli process plant odour information. Females lack a macroglomerular complex. We measured the spatial representation of odours using in-vivo optical recording. We found that: (1) pheromone substances elicited activity exclusively in the MGC. No response was found in female antennal lobes. (2) Plant odours elicited combinatorial activity patterns in the ordinary glomeruli in both males and females. No response was found in the MGC of male moths. (3) A clean air puff often led to activity, in both males and females, suggesting that mechano-sensory information is also processed in the antennal lobe. (4) With an interstimulus interval of 5 or 10 s, strongly activated glomeruli were able to follow the temporal structure of the stimulus, while others lost their phase-locking. Some glomeruli showed "off" responses. These properties were odour dependent. This confirms and extends previous studies, showing the functional significance of the two subsystems for processing olfactory information. Pheromones are coded in a combinatorial manner within the macroglomerular complex, with each glomerulus corresponding to one information channel. Plant odours are coded in an across-glomeruli code in the ordinary glomeruli.     

Odour-genes eovariance and differential investigation of individual odours in the Mus species complex

Previous research using habituation techniques has demonstrated that greater genetic similarity between two individuals is associated with more similarity in the qualities of their individual odours ('odour-genes eovariance'). We assessed odour similarities across species in two pairs of genetically close species within the Mus species complex (M. musculus and M. domesticus; M. spicilegus and M. macedonicus). Subjects treated odours within each species pair as similar compared with an odour from the other species pair. Subjects also treated odours of M. spicilegus males from the same population as similar compared with the odour of M. spicilegus males from a different population. This confirms odour-genes eovariance across species and within populations and also supports previous findings that odour similarities are more salient than specific odour markers. When adult males were presented with odours of females from two different heterospecific species, subjects spent more time investigating the odour from his own species pair than the other species pair, indicating greater interest in the odour of the closer heterospecific and demonstrating that odour-genes eovariance is reflected in behavioural responses to odours. Implications of odour-genes eovariance as a basis for identifying degrees of genetic relatedness of unfamiliar individuals through similarities in individual odours are discussed.     

Concurrent modulation of neuronal and behavioural olfactory responses to sex and host plant cues in a male moth

Mating has profound effects on animal physiology and behaviour, not only in females but also in males, which we show here for olfactory responses. In cotton leafworm moths, Spodoptera littoralis, odour-mediated attraction to sex pheromone and plant volatiles are modulated after mating, producing a behavioural response that matches the physiological condition of the male insect. Unmated males are attracted by upwind flight to sex pheromone released by calling females, as well as to volatiles of lilac flowers and green leaves of the host plant cotton, signalling adult food and mating sites, respectively. Mating temporarily abolishes male attraction to females and host plant odour, but does not diminish attraction to flowers. This behavioural modulation is correlated with a response modulation in the olfactory system, as shown by electro-physiological recordings from antennae and by functional imaging of the antennal lobe, using natural odours and synthetic compounds. An effect of mating on the olfactory responses to pheromone and cotton plant volatiles but not to lilac flowers indicates the presence of functionally independent neural circuits within the olfactory system. Our results indicate that these circuits interconnect and weigh perception of social and habitat odour signals to generate appropriate behavioural responses according to mating state.     

Brief exposure to the odour of a parasitized male alters the subsequent mate odour responses of female mice

Parasites can affect mate choice, with females preferentially selecting parasite-free males. Prior exposure to, or experience with, males has also been suggested to influence mate responses. Here, we examined the effects of an immediate, brief (1 or 15 min) pre-exposure to the urinary and associated odours of either an uninfected male house mouse,Mus domesticus , or a male infected with the natural murine nematode parasite, Heligomosomoides polygyrus, on the subsequent responses of female mice to the odours of infected and uninfected males. Using an odour preference test we found that females displayed a marked overall preference for the odours of uninfected males. Pre-exposure to the odours of a parasitized male decreased female preference for nonparasitized males and increased female preference for parasitized males. This ‘prior male’ effect was evident for both the total odour preference and initial odour choice. Females pre-exposed to the odour of a parasitized male displayed decreased choosiness and made no particular initial choice of a male, while still maintaining an overall preference for the odours of uninfected males. These shifts in initial choice and odour preference were not directly associated with either female stress responses or stress-related odour cues of the males. Our findings show that female mice can distinguish between parasitized and nonparasitized males on the basis of odour and suggest that a brief exposure to the odours of infected males influences females' immediate odour preferences, and their subsequent preference for and choice of males. This rapid, infection-associated ‘prior male’ effect may contribute to the reported variations in female mate preference and choice. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour      

Host feeding experience affects host plant odour preference of the polyphagous leafminer Liriomyza bryoniae

Herbivorous insects use highly specific volatiles or blends of volatiles characteristic to particular plant species to locate their host plants. Thus, data on olfactory preferences can be valuable in developing integrated pest management tools that deal with manipulation of pest insect behaviour. We examined host plant odour preferences of the tomato leafminer, Liriomyza bryoniae (Kaltenbach) (Diptera: Agromyzidae), which is an economically important agricultural pest widespread throughout Europe. The odour preferences of leafminers were tested in dependence of feeding experiences. We ranked host plant odours by their appeal to L. bryoniae based on two‐choice tests using a Y‐tube olfactometer with five host plants: tomato, Solanum lycopersicum Mill.; bittersweet, Solanum dulcamara L.; downy ground‐cherry, Physalis pubescens L. (all Solanaceae); white goosefoot, Chenopodium album L. (Chenopodiaceae); and dead nettle, Lamium album L. (Lamiaceae). The results imply that ranking of host plant odours by their attractiveness to L. bryoniae is complicated due to the influence of larval and adult feeding experiences. Without any feeding experience as an adult, L. bryoniae males showed a preference for the airflow with host plant odour vs. pure air, whereas females did not display a preference. Further tests revealed that adult feeding experience can alter the odour choice of L. bryoniae females. After feeding experience, females showed a preference for host plant odour vs. pure air. Feeding experience in the larval stage influenced the choice by adults of both sexes: for males as well as females reared on bittersweet the odour of that plant was the most attractive. Thus, host feeding experience both in larval and/or adult stage of polyphagous tomato leafminer L. bryoniae influences host plant odour preference by adults.     

Evidence of a Male Sex Pheromone in the Round Goby (Neogobius melanostomus)

The reproductive success of the round goby (Neogobius melanostomus), an invasive fish, may be mediated by the use of pheromones. We hypothesized that reproductive male (RM) round gobies release sex pheromone to which reproductive females (RF) respond. In this study, we compared behavioural and electrophysiological responses of reproductive and non-reproductive female round gobies to conspeci fic males. Results of behavioural experiments in the laboratory showed that RF spent significantly more time near the source of the male odour compared with odours from control water. However, RF did not distinguish between odours from non-reproductive male (non-RM) water and control water. Non-reproductive females (non-RF) were not attracted to odours released from RM or non-RM water. Results of electro-olfactogram (EOG) responses showed that both RF and non-RF discriminated between HPLC fractionated RM and non-RM odours. However, the EOG responses of RF were about eight-fold higher than non-RF exposed to RM odours. These findings confirm that RM round gobies release a pheromone signal that attracts RF. The results of this research may be useful in developing control strategy using natural pheromones to disrupt the reproductive behaviour of the invasive round goby and to curtail its effects on native species. An erratum to this article is available at .     

Larval and adult nutrition effects on blood/nectar choice of Culex nigripalpus mosquitoes

Abstract. The impact of nutritional variables on the development of host-seeking and biting behaviours after emergence by female Culex nigripalpus mosquitoes were studied using air-flow olfactometer and close-range biting assays, respectively. Unfed females failed to develop resting stage ovarian follicles. When offered a bird host in the absence of competing stimuli, sugar-fed mosquitoes were significantly more responsive in both host-seeking and biting than unfed controls. In a choice olfactometer assay using nectar odours (honey scented with artificial apple-blossom oil) versus host odours (a bird), unfed females preferred honey over bird odours except when honey odour was weak. After sucrose feeding, females switched from honey to bird preference. This change in behaviour was accompanied by significant accumulation of lipid and by follicular growth to the resting stage. Elevation of host responsiveness after sugar feeding was reversible; starvation ultimately resulted in females preferring honey over bird odours. When the larval diet was restricted by crowding, the wing-length and total lipid of resultant adult females were reduced. Although differences were subtle, unfed bird-responding females tended to have longer wings and more lipid than their honey-responding counterparts.     

Herbivore-induced emissions of maize volatiles repel the corn leaf aphid, shape Rhopalosiphum maidis

When maize plants, Zea mays L., are mechanically damaged and the damaged sites are treated with caterpillar regurgitant, the plants will release a specific blend of volatiles. It is known that these volatiles can be attractive to natural enemies of herbivores. We hypothesise that the plant volatiles constitute part of the induced plant defence and that herbivores will be affected by the odours as well. In laboratory and semi-field studies this hypothesis was tested for the aphid Rhopalosiphum maidis (Fitch) (Rhynchota, Sternorrhyncha, Aphididae).In a Y-tube olfactometer significantly more aphids chose the odour of healthy, undamaged maize seedlings when tested against clean air or plants treated with regurgitant. Clean air was chosen more often when tested next to the odour of treated plants. This apparently repellent effect of the odour of treated plants was significant for winged aphids, but not for the wingless aphids.In field experiments aphids were released in the centre of circles of eight potted maize plants. Four plants in each circle were damaged and treated with caterpillar regurgitant while the other plants were left unharmed. At different intervals after aphid release, the number of aphids was counted on each plant. Significantly fewer winged and wingless aphids were found back on treated plants than on healthy plants.We suggest that herbivores may be repelled by the odours because they could indicate that: 1) the plant has initiated the production of toxic compounds; 2) potential competitors are present on the plant; 3) the plant is attractive to parasitoids and predators. Aphids may be particularly sensitive to induced maize volatiles because one of the major compounds emitted by the plant is (E)--farnesene, which is a common alarm pheromone for aphids. Collections and analyses of the odours emitted by crushed R. maidis confirmed that it too emits (E)--farnesene when stressed. The results are discussed in context of plant defence strategies and their possible exploitation for the control of pest insects.     

The effect of repellents Ocimum forskolei and deet on the response of Anopheles stephensi to host odours

The behavioural response of Anopheles stephensi Liston (Diptera: Culicidae) to incubated host odours (from human and goat) and to human odour in combination with a repellent plant, Ocimum forskolei (Labiatae), or deet (N, N, diethyl-toluamide) (20%) was tested in a dual-port olfactometer. An. stephensi was significantly attracted to both host odours compared with air alone, but showed no preference when given a choice between both host odours simultaneously. In choice tests, the addition of deet to human odour did not significantly divert mosquitoes to human odour alone, but did divert them to goat odour. O. forskolei combined with human odour diverted mosquitoes to goat or human odour alone. Combinations of human odour and O. forskolei, and human odour and deet were both as unattractive as air alone, and attracted mosquitoes equally when offered simultaneously. The results indicate that use of O. forskolei as a repellent would be beneficial in reducing vector biting if used in communities in areas with partially zoophilic mosquito species such as An. stephensi, and where animals are present.     

Foraging and mate-finding in the silver Y moth, Autographa gamma (Lepidoptera: Noctuidae) under the risk of predation

Animal foraging and reproductive behaviour is influenced by other simultaneous demands such as predator avoidance. The trade-offs between these demands may depend on sex or mating experience. This study demonstrates that the olfactory-mediated foraging and mate-seeking behaviours in the silver Y moths, Autographa gamma , are affected by auditory cues mimicking their bat predators. Both males and females changed their foraging behaviour under simulated predation risk. Fewer moths reached the odour source following sound stimulation and the time to find the odour source increased by up to 250%. However, there were no significant differences between male and female ability to reach the plant odour source or the duration of the flight towards the source when stimulated with ultrasound. Hence females are not more cautious than males when observed in the same behavioural context. Risk-taking in males was independent of whether they were flying toward a flower odour or sex pheromones having equal attractive value. This indicates that the trade-off between olfactory and acoustic cues is independent the type of odour. Mated females were not as strongly affected by sound as non-mated, indicating that flower odours have a higher adaptive value for mated females, suggesting that some processes following mating experience influence the trade-off between flower odours and simulated bat sounds.