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.     

Mixed blends of herbivore-induced plant volatiles and foraging success of carnivorous arthropods

Food webs are overlaid with infochemical webs that mediate direct and indirect interactions. Behavioural ecologists have extensively documented that carnivorous arthropods exploit herbivore-induced plant volatiles during foraging for herbivorous arthropods. Most studies on the role of infochemicals in multitrophic interactions have been conducted against an odour-free background, although field studies show that carnivores also use herbivore-induced plant volatiles under more complex conditions. Here we investigated the effect of mixing the blends of volatiles emitted by two plant species on the foraging behaviour of the predatory mite Phytoseiulus persimilis . This was done in an olfactometer under laboratory conditions and in a semi-field setup under greenhouse conditions. The olfactometer setup ensured directed mixing of the two odour blends, while odour mixing in the greenhouse setup was much less controlled and resulted from diffusion. In 4 out of 5 olfactometer experiments the behaviour towards volatiles from spider-mite ( Tetranychus urticae ) infested Lima bean plants was not affected by mixing with volatiles from caterpillar ( Pieris brassicae ) infested Brussels sprouts plants. In the fifth olfactometer experiment the response shifted significantly towards the volatiles from infested Lima bean leaves without volatiles from infested cabbage leaves. In the greenhouse setup no effect of infested cabbage plants or their volatiles on the location of spider-mite infested bean plants was recorded. The two odour blends used in this study, i.e. those from spider-mite infested Lima bean leaves and from caterpillar-infested Brussels sprouts plants, are very different and there is no overlap in compounds that are known to attract the predators. The results are discussed in the context of other types of odour-blend mixing and the effects on food web interactions.     

Anthocorid predators learn to associate herbivore-induced plant volatiles with presence or absence of prey

We investigated how the plant‐inhabiting, anthocorid predator, Anthocoris nemoralis, copes with variation in prey, host plant and associated herbivore‐induced plant volatiles and in particular whether the preference for these plant odours is innate or acquired. We found a marked difference between the olfactory response of orchard‐caught predators and that of their first generation reared on flour moth eggs in the laboratory, i.e. under conditions free of herbivory‐induced volatiles. Whereas the orchard‐caught predators preferred odour from psyllid‐infested pear leaves, when offered against clean air in a Y‐tube olfactometer, the laboratory‐reared first generation of (naive) predators did not. The same difference was found when a single component (methyl salicylate) of the herbivore‐induced plant volatiles was offered against clean air. After experiencing methyl salicylate with prey, however, the laboratory‐reared predators showed a pronounced preference for this volatile. This acquired preference did not depend on whether the volatile had been experienced in the juvenile period or in the adult phase, but it did depend on whether it had been offered in presence or absence of prey. In the first case, they were attracted to the plant volatile in subsequent olfactometer experiments, but when the volatile had been offered during a period of prey deprivation, the predators were not attracted. We conclude that associative learning is the most likely mechanism underlying acquired odour preference.     

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.     

The effect of rearing history and aphid density on volatile‐mediated foraging behaviour of Diaeretiella rapae

1. Parasitoid females foraging for hosts rely on cues derived from the insect host, the host plant and/or their interaction, and all of these can be learned during the immature and adult stages. 2. The present study investigated the importance of rearing history on foraging behaviour of Diaeretiella rapae, an endoparasitoid often associated with aphids feeding on brassicaceous plant species. 3. Parasitoids were reared on one of the four possible combinations, comprising two brassicaceous host plant species, Brassica nigra or Raphanus sativus, and two aphid species Brevicoryne brassicae or Myzus persicae. These parasitoids were tested in a Y‐tube olfactometer and given the choice between volatiles emitted by an aphid‐infested plant (25 or 100 aphids per plant) and an uninfested control plant. The parasitoid's responses were compared when offered: (i) the same plant–aphid combination as the one on which it had been reared; (ii) the same host plant infested with the alternative aphid species; or (iii) an alternative plant with the alternative aphid species. 4. Aphid density did affect the behavioural responses to the various odour sources, but rearing history did not. Diaeretiella rapae only preferred aphid‐induced to non‐induced plant volatiles at low aphid infestation level, whereas they did not discriminate between volatiles at high aphid infestation level. 5. It is concluded that aphid‐induced volatiles of brassicaceous plants play an important role during host habitat location, but seem less important for parasitoids to locate the aphid host itself. The data are discussed in the light of manipulation of host plant defences by aphids.     

Innate olfactory responses of female and male parasitoid Apanteles taragamae Viereck (Hymenoptera: Braconidae) toward host plant infested by the cucumber moth Diaphania indica Saunders (Lepidoptera: Crambidae)

The behavioural responses of Apanteles taragamae, a larval parasitoid of the cucumber moth Diaphania indica, to the volatiles of cucumber plants was investigated in a four-arm olfactometer. Females and males were given a choice between several odour sources that included (1) clean air, (2) uninfested, (3) host-infested, and (4) mechanically damaged cucumber plants. Females and males showed different preferences for volatiles emanating from these plants. Females responded significantly longer to the volatiles from uninfested plants than clean air, and to host-infested plants than uninfested plants. There were no significant differences in female responses to the volatiles from mechanically damaged and uninfested plants. Males responded significantly longer to clean air rather than uninfested plants. The volatiles from both uninfested and host-infested cucumber plants may play important roles in host habitat location of A. taragamae females.     

Odour masking of tomato volatiles by coriander volatiles in host plant selection of Bemisia tabaci biotype B

The silverleaf whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is one of the most important pest insects in tomato crop systems worldwide. It has been previously demonstrated that intercropping tomato [Solanum lycopersicum L. Mill. (Solanaceae)] with coriander [Coriandrum sativum L. (Apiaceae)] reduces the incidence and severity of damage caused by B. tabaci. However, it is not yet known how coriander affects the insect′s behaviour. We evaluated the attractiveness of tomato constitutive volatiles to B. tabaci and what effect coriander constitutive volatiles have on the insect′s behaviour. To this end, we conducted three bioassays in a multiple‐choice four‐arm olfactometer (‘×’ type), measuring B. tabaci behaviour when offered tomato and coriander constitutive volatiles presented alone as well as together. We also evaluated the colonisation and establishment of B. tabaci in experimental plots with only single tomato plants and tomatoes intercropped with coriander in a greenhouse. Bemisia tabaci males and females recognised tomato constitutive volatiles as a positive stimulus (kairomonal effect), indicating that semiochemicals from this plant can play an important role in the insect’s host plant selection. Coriander constitutive volatiles reduced the attractiveness of tomato volatiles but no repellency to these volatiles was observed. Greater numbers of adults and nymphs of B. tabaci per plant were observed in tomato monoculture plots than in tomato intercropped with coriander. We suggest that coriander constitutive volatiles have an odour masking effect on tomato volatiles, thus interfering in the host plant selection of B. tabaci.     

The effects of non-host plant essential oil volatiles on the behaviour of the pollen beetle Meligethes aeneus

The use of semiochemicals for manipulation of the pollen beetle Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae) is being investigated for potential incorporation into a push‐pull control strategy for this pest, which damages oilseed rape, Brassica napus L. (Brassicaceae), throughout Europe. The response of M. aeneus to non‐host plant volatiles was investigated in laboratory assays to establish whether they have any effect on host plant location behaviour. Two approaches were used. First a novel, moving‐air bioassay using air funnels was developed to compare the response of M. aeneus to several non‐host plant essential oils. The beetles avoided the host plant flowers in the presence of non‐host volatiles, suggesting that M. aeneus uses olfactory cues in host location and/or acceptance. The results were expressed as ‘repellency values’ in order to compare the effects of the different oils tested. Lavender (Lavendula angustifolia Miller) (Lamiaceae) essential oil gave the highest repellency value. In addition, a four‐arm olfactometer was used to investigate olfactory responses, as this technique eliminated the influence of host plant visual and contact cues. The attraction to host plant volatiles was reduced by the addition of non‐host plant volatiles, but in addition to masking the host plant volatiles, the non‐host volatiles were avoided when these were presented alone. This is encouraging for the potential use of non‐host plants within a push‐pull strategy to reduce the pest colonisation of crops. Further testing in more realistic semi‐field and field trials is underway.     

Odour‐mediated orientation behaviour of the lily beetle Lilioceris lilii

The lily beetle Lilioceris lilii (Scopoli) (Coleoptera: Chrysomelidae) feeds on Lilium, Fritillaria and Cardiocrinum plants and is a serious pest in gardens and amenity plantings in parts of Northern Europe and North America. In the present study, the odour‐mediated behaviour of L. lilii is investigated by behavioural bioassays using a linear‐track olfactometer. The behavioural responses of L. lilii to hosts and conspecifics are, at least in part, odour‐mediated and the responses differ with respect to the physiological (reproductive) state of the adult beetle (i.e. pre‐ or post‐diapause). Significantly more diapaused female L. lilii move into air streams containing the odour of intact host plants than into clean air, and move into air streams containing odour of host plants and beetles combined in preference to odour from manually‐damaged host plants. Diapaused females also move into air streams containing odours from intact plants over those from larval‐infested plants. Pre‐diapause males move into the air streams of intact plants rather than L. lilii‐infested plants. Pre‐diapause females show no significant response in any experiment. The data indicate that the odour‐mediated responses of L. lilii are consistent with those known for other chrysomelids that produce a male aggregation pheromone to which reproductive individuals of both sexes respond.     

The responses of the cabbage seed weevil (Ceutorhynchus assimilis) to the odour of oilseed rape (Brassica napus) and to some volatile isothiocyanates

The responses of the cabbage seed weevil,Ceutorhynchus assimilis Payk. (Coleoptera: Curculionidae) to volatiles from oilseed rape (Brassica napus L.) were tested using a linear track olfactometer. Weevils were attracted towards the odour of rape during a short period before diapause and for most of their postdiapause life. Odours from both the flowering and green parts of the plant were attractive, but the odour of a non-host plant (Bellis perennis L.) was not. An entrainment extract of flowering rape volatiles in pentane was attractive, but significantly less so than the odour of flowering rape itself. Attraction was found to 3-butenyl and 4-pentenyl isothiocyanate (NCS), but not to 2-phenylethyl isothiocyanate. A mixture of the three NCSs was more attractive than the individual NCSs. However, the concentration required to produce a response was still relatively high. Isothiocyanates, along with other volatiles from rape, probably play a role in host plant recognition by the seed weevil.     

Volatiles from host plant brinjal attract the brinjal Fruit and Shoot Borer -Leucinodes orbonalis Guenee

Brinjal Fruit and Shoot Borer- Leucinodes orbonalis Guenee is a major insect pest on brinjal- Solanum melongena worldwide. An effective strategy used in developing pest controlling agents is the synergism between insect pheromones and host plant volatiles, which can increase the attraction of insect pest. The present study was aimed at investigating the chemical constituents and attractant effects of the volatiles extracted from different parts of the host plant brinjal on the behavior of adult L. orbonalis. Bioassay using Y-shaped olfactometer revealed that the one-day old virgin female, gravid female and male insects respond positively to the host plant volatiles extracted from fruits, leaves and shoots but not to that of flowers. It was shown that the gravid females were significantly attracted to all three volatiles (p < 0.05). Bioassay using X-shaped olfactometer identified that all three types of insects highly preferred the volatiles from fruits (p < 0.05). Gas chromatography-mass spectrometry analysis of volatiles indicated that brinjal plant produces volatile secondary metabolites, which include 2,2′-(Ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) dibenzoate (12.11%), 3,7-dimethylocta-1,6-dien-3-ol (22.38%), Benzyl alcohol (22.9%) and Benzyl alcohol (27.06%) as major constituents from fruits, shoots, leaves and flowers respectively. Responses of insects to the volatiles from host plant in the absence of visual cues direct us to focus on the importance of host plant volatiles to locate the plant. Results of this study emphasize the major role that host plant volatiles play in the attraction of insect pests towards the plant.     

Habitats as Complex Odour Environments: How Does Plant Diversity Affect Herbivore and Parasitoid Orientation?

Plant diversity is known to affect success of host location by pest insects, but its effect on olfactory orientation of non-pest insect species has hardly been addressed. First, we tested in laboratory experiments the hypothesis that non-host plants, which increase odour complexity in habitats, affect the host location ability of herbivores and parasitoids. Furthermore, we recorded field data of plant diversity in addition to herbivore and parasitoid abundance at 77 grassland sites in three different regions in Germany in order to elucidate whether our laboratory results reflect the field situation. As a model system we used the herb Plantago lanceolata, the herbivorous weevil Mecinus pascuorum, and its larval parasitoid Mesopolobus incultus. The laboratory bioassays revealed that both the herbivorous weevil and its larval parasitoid can locate their host plant and host via olfactory cues even in the presence of non-host odour. In a newly established two-circle olfactometer, the weeviĺs capability to detect host plant odour was not affected by odours from non-host plants. However, addition of non-host plant odours to host plant odour enhanced the weeviĺs foraging activity. The parasitoid was attracted by a combination of host plant and host volatiles in both the absence and presence of non-host plant volatiles in a Y-tube olfactometer. In dual choice tests the parasitoid preferred the blend of host plant and host volatiles over its combination with non-host plant volatiles. In the field, no indication was found that high plant diversity disturbs host (plant) location by the weevil and its parasitoid. In contrast, plant diversity was positively correlated with weevil abundance, whereas parasitoid abundance was independent of plant diversity. Therefore, we conclude that weevils and parasitoids showed the sensory capacity to successfully cope with complex vegetation odours when searching for hosts.     

Aggregation pheromone in Sternechus subsignatus (Coleoptera: Curculionidae): olfactory behaviour and temporal pattern of emission

The Brazilian soybean stalk weevil, Sternechus subsignatus Boehman (Coleoptera: Curculionidae), is a pest of economic importance in many regions of Brazil. Volatiles from both sexes of S. subsignatus were collected by aeration and the behavioural response of males and females was evaluated using a Y‐olfactometer. The results obtained demonstrate that the communication in S. subsignatus is mediated by aggregation pheromone as both sexes were attracted to host plant (HP) volatiles, and this attraction was increased by the addition of male volatiles. At least five male‐specific compounds (1–5) were detected in the chromatographic analysis, providing chemical support to the behavioural data. Release of these volatiles is dependent on the presence of the HP as the amount of compounds differs significantly when volatiles are collected from weevils with or without access to food. The release takes place mainly during photophase, showing a peak between 4 and 6 h after its beginning, which is also a peak of the insect activity in the field. Studies are underway to elucidate the structures of these putative aggregation pheromone components, in order to evaluate their biological activity under laboratory and field conditions.     

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.     

Olfactory cues used in host‐habitat location and host location by the parasitoid Cotesia urabae

There is a growing body of evidence that many hymenopteran parasitoids make use of olfaction as the primary mechanism to detect and locate hosts. In this study, a series of bioassays was conducted to investigate the orientation behaviour of the gum leaf skeletonizer larval parasitoid Cotesia urabae Austin & Allen (Hymenoptera: Braconidae) in both Y‐tube and four‐arm olfactometers. In a Y‐tube olfactometer, male C. urabae were attracted only to virgin conspecific females. Host‐plant leaves, damaged leaves, host larvae, and host larvae feeding on leaves were highly attractive to female C. urabae, whereas host frass and conspecific males were not. The multiple‐comparison bioassay conducted in a four‐arm olfactometer clearly indicates that C. urabae females were significantly more attracted to the host Uraba lugens Walker (Lepidoptera: Nolidae) larvae feeding on Eucalyptus fastigata H Deane & Maiden (Myrtaceae) leaves than to any other of the odour sources tested. The results of this study show that C. urabae individuals responded to chemical cues specific to the host plant and target host insect, and support hypotheses that unreliable cues are not utilized for host location by specific natural enemies.     

RESPONSE OF PARASITOID MICROPLITIS MEDIATOR TO PLANT VOLATILES IN AN OLFACTOMETER*

Abstract The behavioral responses of Microplitis mediator were measured in a four‐armed olfactometer. Leaves extract of the suitable host plant of its insect host, cotton, elicited higher olfactory responses; while leaves extract of the less suitable host plant of its insect host, tomato or tobacco, elicited moderate responses. Volatiles from the least suitable host plant of its insect host, hot pepper, elicited the lowest responses. The different preferences to four plant leaves extracts suggested that M. mediator could discriminate between suitable and less suitable host plants of its insect host by semiochemicals. Green leaf volatiles (GLVs), cis‐3‐hexen‐1‐yl acetate, hexanal, and nonan‐1‐ol, are the most attractive compound among the 7 tested compounds. The parasitoids preferred host‐damaged plant to undamaged plant, suggesting that they were able to distinguish between host and non‐host of their insect host through varying chemical blends. Experience with cotton leaves extract prior to experiment enhanced their responses to the same odor.     

Role of the plant–conspecific complex in host location and intra-specific communication of Lygus rugulipennis

Abstract.  Wind tunnel and vertical open Y-shaped olfactometer studies are used to test whether volatile cues from the host plant ( Vicia faba ), from conspecific bugs, and from a plant–conspecifics combination, would elicit behavioural responses in mated males and females of Lygus rugulipennis . In the olfactometer, females move towards volatiles from healthy plants but they do not respond to volatiles released by oviposition- and/or feeding-damaged plants without conspecifics, nor to conspecifics alone. Both in the wind tunnel and olfactometer, females respond to volatiles emitted by the plant–insect complex. By contrast, in the wind tunnel, both sexes move significantly towards damaged host plants, even if the presence of conspecifics on these plants enhances only the female response. However, the presence of eggs from conspecifics on host plants reduces the responses of both sexes in the wind tunnel. Finally, males, as well as females, are less responsive to conspecifics alone compared with damaged plants, especially when conspecifics are present on the host plants (host plant– Lygus complex). The results suggest that volatiles emitted by plants and conspecifics influence L.   rugulipennis behaviour, giving information to both sexes on the presence of suitable host plants that have been colonized by other conspecifics acting as pioneers, or providing information on the presence of an already exploited host plant (presence of eggs), thus preventing competition. Males also can use this information to increase the probability of encountering mature females.     

Host location by chemical stimuli in Bracon hylobii (Ratzeburg) (Hymenoptera: Braconidae), a larval parasitoid of Hylobius abietis (L.) (Coleoptera: Curculionidae)

The Large Pine Weevil, Hylobius abietis, is one of the most important pests in European coniferous forests. The ectoparasitoid Bracon hylobii is known to cause around 50% larval mortality. Chemical volatiles are among the most important stimuli involved in the host location. In the present study, a 4-arm olfactometer and a 2-choice wind-tunnel were used to test the behavioural responses of B. hylobii females to four odour sources from the host-plant/host-insect complex. For each odour, the behaviour of twenty virgin females was individually recorded in an olfactometer, whereas twenty virgin and twenty mated females were individually tested in a wind-tunnel. The following behavioural parameters were analysed: a) time spent walking; b) time spent stopped; c) total time spent in each arm; d) total speed; e) net speed; f) the rate of turning per unit distance walked; g) rate of turning per second. Comparison between odours shows clear differences in the behavioural parameters analysed. In particular, the bark disc/larva complex is the only odour source able to elicit behavioural responses of both naive and mated females. As strong responses were obtained only using experienced females, this suggests that previous searching activities increase the ability of B. hylobii to find its host.     

An airflow olfactometer for measuring olfactory responses of hymenopterous parasitoids and other small insects

ABSTRACT. A new type of airflow olfactometer is described, and results given of experiments using it to measure behavioural olfactory responses of hymenopterous parasitoids. Compared with Y-tube olfactometers it shows several advantages. In its exposure chamber four separate abutting odour fields are presented so that the test insect can readily enter and re-enter them. More than one odour (or different concentrations of one odour) can be tested at the same time, thereby providing complex preference test situations. The various behavioural measures that can be assessed in the apparatus are examined and discussed.     

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.