Fight or flight? A geographic mosaic in host reaction and potency of a chemical weapon in the social parasite <Emphasis Type="Italic">Harpagoxenus sublaevis</Emphasis>

Ant social parasites use chemical warfare to facilitate host colony takeover, which is a critical but recurring step in their life cycle. Many slave-making ants use the secretion of the Dufour gland to manipulate host behaviour during parasitic nest foundation and slave raids. Harpagoxenus sublaevis applies this chemical weapon onto defending Leptothorax host workers, which elicits deadly fights amongst them. Host species are expected to evolve counter-adaptations against this behavioural manipulation and in this study we investigated the geographic structure of this co-evolving trait. We compared the effectiveness of the parasitic gland secretion from different H. sublaevis populations in host colonies from various sites and analysed the occurrence of local adaptation. The two host species L. muscorum and L. acervorum generally showed different responses to the parasites’ chemical weapon: L. acervorum attacked nestmates treated with Dufour gland secretion, while L. muscorum workers fled. Flight, instead of intraspecific fights, is an adaptive host reaction as it results in fewer host fatalities during raids. Beside interspecific host differences, we found a geographic mosaic of host resistance: parasites from a German population strongly manipulated the behaviour of both sympatric Leptothorax populations. Russian or Italian hosts instead did not react with intracolonial aggression, but fled when confronted with the gland secretion of their sympatric parasite. Not only variation in host resistance explains differences in the effectiveness of the parasitic gland secretion but also interpopulational differences in its chemical composition, which were revealed by gas chromatography and mass spectrometry.     

Disease resistance in a weaver ant, <Emphasis Type="BoldItalic">Polyrhachis dives</Emphasis>, and the role of antibiotic-producing glands

Parasites represent one of the main threats to all organisms and are likely to be particularly significant for social animals because of the increased potential for intragroup transmission. Social animals must therefore have effective resistance mechanisms against parasites and one of the most important components of disease resistance in ants is thought to be the antibiotic-producing metapleural gland. This gland is ancestral in ants, but has been lost secondarily in a small number of species. It is unknown whether these evolutionary losses are due to a reduction in parasite pressure or the replacement of the gland’s function with other resistance mechanisms. Here we used the generalist entomopathogenic fungus Metarhizium to compare the disease resistance of a species of a weaver ant, Polyrhachis dives, which has lost the metapleural gland, with that of the well-studied leaf-cutting ant Acromyrmex echinatior and two other ant species, Myrmica ruginodis and Formica fusca, all of which have metapleural glands. The P. dives weaver ants had intermediate resistance when kept individually, and similar resistance to A. echinatior leaf-cutting ants when kept in groups, suggesting that the loss of the metapleural gland has not resulted in weaver ants having reduced disease resistance. P. dives weaver ants self-groomed at a significantly higher rate than the other ants examined and apparently use their venom for resistance, as they had reduced resistance when their venom gland was blocked and the venom was shown in vitro to prevent the germination of fungal spores. Unexpectedly, the leaf-cutting ant A. echinatior also had reduced resistance to Metarhizium when its venom gland was blocked. It therefore appears that the evolutionary loss of the metapleural gland does not result in reduced disease resistance in P. dives weaver ants, and that this at least in part may be due to the ants having antimicrobial venom and high self-grooming rates. The results therefore emphasise the importance of multiple, complementary mechanisms in the disease resistance of ant societies.     

Behavioural evidence for the involvement of Dufour’s gland secretion in nestmate recognition in the social wasp Polistes dominulus (Hymenoptera: Vespidae)

The results of recent studies by our group have suggested that in social wasps of the genus Polistes the Dufour’s gland is involved in kin recognition. In fact, the same hydrocarbons occurring on the cuticle are found in the gland secretion, and in P. dominulus the composition of the glandular secretion is more similar in foundresses belonging to the same colonies than in heterocolonial foundresses. In this study, P. dominulus colonies were experimentally presented with previously treated dead conspecific females. These lures had been deprived of their epicuticular lipids and coated with epicuticular or Dufour’s gland secretion extracts from females hetero- or homocolonial with respect to the tested colonies. The behaviour of the colonies towards these lures indicates that, like the epicuticular lipids, the Dufour’s gland secretion is involved in nestmate recognition. Received: 14 August 1995/Accepted after revision: 5 November 1995     

Structure and function of Dufour gland pheromones from the crazy ant <Emphasis Type="Italic">Paratrechina longicornis</Emphasis>

Summary. Dufour glands of Paratrechina longicornis were analyzed chemically using GC-MS techniques. The glands contained twelve alkanes between C₉ and C₂₀ chain length with undecane and tridecane as the main components, six alkenes (1- and 4-undecene, 1-, 4-, and 6- tridecene, and heptadecadiene), two alkyl formates (C₁₁ and C₁₃), and saturated and polyunsaturated acids. Many of the alkanes and alkenes were behaviorally active causing a short lasting attraction of ants with different intensity. Detection of the major gland compound undecane in recruitment trails by in vivo SPME sampling provided evidence for its use as a recruitment pheromone. Both poison gland (formic acid) and Dufour gland (undecane) contents were detected on SPME fibers which had been attacked by the ants.     

Trail-following behaviour in two Aphaenogaster ants

In the course of our studies on the chemical ecology of the widely distributed Mediterranean ant Aphaenogaster senilis, we found that trail following is triggered by extracts of the poison gland and Dufour’s gland. To assess the specificity of the trail pheromone, we examined whether a cross-reaction exists between trails of A. senilis and the closely related species A. iberica. Specificity seemed to differ amongst these two species, because workers of A. senilis did not follow trails of A. iberica, whereas the latter followed trails made by both species. Chemical analyses of the glandular contents reveal that Dufour’s glands of both species contain mainly alkanes and alkenes exhibiting species-specific profiles. However, differences in the poison gland content of the two species were dramatic, with A. senilis showing high amounts of alkaloids that were completely absent in A. iberica.     

The role of the queen mandibular gland pheromone in honeybees (<Emphasis Type="Italic">Apis mellifera</Emphasis>): honest signal or suppressive agent?

Queen pheromones interfere with worker reproduction in social insects. However, there is still an unresolved question as to whether this pheromone acts as an “honest” signal for workers, giving a reliable indication of the queen’s reproductive value, or as a suppressive agent, inhibiting worker reproduction independent of the queen’s reproductive capacity. In honeybees (Apis mellifera), the queen’s mandibular gland secretion, a mix of fatty acids and some aromatic compounds, is crucial for regulating the reproductive division of labor in the colony inhibiting ovary development in workers. We quantified the mandibular gland secretions of virgin, drone-laying, and naturally mated queens using gas chromatography to test whether the queens’ mating, ovary activation, or the reproductive value for workers correlated with the composition of the secretion. Although the absolute amounts of the “queen substance” 9-oxo-2(E)-decenoic acid (9-ODA) were similar among the three groups, the proportions of 9-ODA decreased with increasing reproductive quality. Furthermore, the ratios of queen to worker compounds were similar in all three treatment groups, irrespective of the reproductive capacity. A multivariate analysis including all six compounds could not separate drone-laying queens from naturally mated ones, both with active ovaries but only the latter ensuring colony survival. We suggest that the mandibular gland pheromones are unlikely to function as reliable indicators of queen reproductive value and rather operate as an agent to suppress worker reproduction. This does not exclude the possibility that other “honest” pheromone signals exist in the honeybee colony, but these would have to arise from other semiochemicals, which could be produced by both the queen and the brood.     

Learning,retention and coding of nest-associated visual cues by the Australian desert ant, <Emphasis Type="Italic">Melophorus bagoti</Emphasis>

A variety of social insects use visual cues for homing. In this study, we examine the possible factors affecting the learning and retention of nest-associated visual cues by the Australian desert ant Melophorus bagoti and the manner in which such cues are encoded by foraging ants. We placed four prominent cylindrical landmarks around a nest and trained foragers from that nest to a food source. Ants were tested with the landmark array in a distant testing field after (1) a known number of exposures to the landmarks (1, 3, 7 or 15 trials, spread over a period of 1 day, 2 days or ≥3 days) and (2) after a known period of delay (0, 24, 48, 96 or 192 h). The results show that a combination of an increase in training trials and an increase in number of training days affected the acquisition of landmark memory. Moreover, once the landmarks were learnt, they became a part of long-term memory and lasted throughout the ants’ foraging lifetime. To examine visual cue encoding behaviour, ants trained under similar conditions for 4 days were tested with (1) an identical landmark array, (2) landmarks of the same size used in training, but placed at twice the distance from each other, and (3) landmarks whose dimensions were doubled and placed at twice the distance from each other. In conditions (1) and (3), the ants searched extensively at the centre of the four landmarks, suggesting that, similar to the Saharan ant (genus Cataglyphis) and the honeybee, M. bagoti too uses a snapshot to match the view of the landmarks around the nest. But contrary to the snapshot model, in condition (2), the ants did not search extensively at the centre of the landmarks, but searched primarily 0.5 m from the landmark, the distance from each landmark to the nest during training. We discuss how various search models fare in accounting for these findings.     

“Anting” in Blue Jays: evidence in support of a food-preparatory function

Summary.  Anting, the plumage-dipping behavior to which ants (mostly formicines) are commonly subjected by birds (mostly passerines), is shown in tests with hand-raised Blue Jays (Cyanocitta cristata) and the ant Formica exsectoides to be instinctive: the birds displayed typical renditions of the behavior on the first occasion that they encountered ants. Evidence is presented supportive of the view that anting is a strategy by which birds render ants fit for ingestion. Formicine ants are ordinarily protected by their formic acid-containing spray. Being wiped into the bird’s plumage causes them to discharge that spray, without harm to the bird, to the point of almost total emptying of the glandular sac in which the secretion is stored. The ants are therefore essentially secretion-free by the time they are swallowed. Further evidence indicates that it is the ant’s possession of the acid sac that triggers the anting behavior in the bird. If F. exsectoides are surgically deprived of their acid sac, they are eaten by the birds without first being subjected to anting. Data are also presented indicating that the ant’s crop, which is especially capacious in formicines (its contents may amount to over 30% of the formicine’s mass), and which appears to survive the anting procedure intact, constitutes, at least when laden, a valuable component of the trophic package that the bird accesses by anting.     

Defence effectiveness of easy bleeding sawfly larvae towards invertebrate and avian predators

Summary. Easy bleeding is a phenomenon discovered in some tenthredinid insects which possess a particularly low mechanical resistance of the integument, leading under mechanical stress to haemolymph exudation. It has a defensive effect against ants and wasps through harmful plant compounds which are sequestered in the haemolymph. Here we describe etho-ecological and some chemical aspects of the defence of easy bleeders and specify the range of predators to which easy bleeding might be effective. Beside a high haemolymph deterrence associated with low integument resistance across sawfly species, we also detected toxicity of the haemolymph of some species to workers of the ant Myrmica rubra. The behaviour of easy bleeders is to move slowly and, once disturbed, to become motionless, thereby probably impeding the tendency of a predator to attack. This behaviour had no beneficial effect for easy bleeders when attacked by the predatory bug Podisus maculiventris. Bugs could successfully and without harm prey on sawfly larvae without evoking easy bleeding. For the easy bleeder Athalia rosae, host plants with different secondary metabolite profiles, and, consequently, changes in haemolymph chemistry only slightly affected the feeding behaviour of the bugs. To test the effectiveness of easy bleeding towards a vertebrate predator, easy bleeders were offered to birds, Sturnus vulgaris. The body colouration of the sawfly larvae was of prime importance in determining the predators response when testing birds in a group. It is likely that easy bleeding is a defence strategy directed primarily towards foraging insects with biting-chewing mandibles and that it is much less active towards predatory insects with piercing-sucking mandibles as well as birds. The involvement of chemical and/or physical cues in the strategy is discussed with respect to these types of predators.     

Social facilitation of exploration in mosquitofish (<Emphasis Type="Italic">Gambusia holbrooki</Emphasis>)

The social environment can exert a powerful influence on the expression of an individual’s behaviour patterns. For example, social facilitation occurs where individuals are more likely to express a given behaviour, or express it a greater rate, in the presence of conspecifics. Social facilitation is partly driven by an individual’s perception of risk, which is a function both of the size of its social group and the information that it gathers relating to predator activity and risk. Here I tested the effects of social group size (one, two, four, eight or 16 fish) and the presence of ‘social’, ‘predation’ or ‘neutral’ chemical cues (derived respectively from live conspecifics, injured conspecifics or a blank water control) on the exploratory behaviour of juvenile mosquitofish (Gambusia holbrooki) in a novel environment. Focal fish in larger groups explored a greater proportion of the arena during the course of the experiment, demonstrating social facilitation of exploration. After 4 h in the arena, focal fish in all group sizes showed significantly reduced swimming activity, suggesting that the initial faster swimming activity of fish on entry to the arena may be in response to the motivation to explore. The presence of predation cues in the environment had the effect of reducing exploratory behaviour across groups in a novel environment, whereas social cues had no effect on exploratory behaviour. Taken as a whole, these results suggest that there is a high degree of context dependency in the expression of exploratory behaviour, with a strong influence of both the presence of conspecifics and cues relating to potential danger.     

Offensive and defensive sperm competition roles in the dung beetle Onthophagus taurus (Coleoptera: Scarabaeidae)

Sperm competition is predicted to generate opposing selection pressures on males. On one hand, selection should favour ‘defensive’ adaptations that protect a male’s ejaculate from displacement, while, on the other hand, selection should favour ‘offensive’ adaptations that overcome paternity assurance mechanisms of rivals. Here, we use the sterile male technique to assess sperm precedence when a male dung beetle Onthophagus taurus mates in both a defensive (first male) and an offensive (second male) role. Significant variation in a male’s sperm precedence (both P ₁ and P ₂) was detected, and an individual’s defensive (P ₁) and offensive (P ₂) abilities were positively correlated. Thus, it appears that sexual selection simultaneously selects for ‘defensive’ and ‘offensive’ adaptations in O. taurus. We discuss a variety of male traits in O. taurus that potentially contribute to a male’s ability to be successful when mating in an ‘offensive’ and a ‘defensive’ role.     

Experimental support for the cost–benefit model of lizard thermoregulation

According to Huey and Slatkin’s [Q Rev Biol 51:363–384, 1976] cost–benefit model of behavioural thermoregulation, lizards should adjust their thermoregulatory strategy between active thermoregulation and thermoconformity (no thermoregulation) according to the costs (time and energy spent thermoregulating, exposure to predators), benefits (optimised physiological performance) and thermal quality of environment associated with a given situation. However, Gilchrist’s [Am Nat 146:252–270, 1995] model of thermal specialisation suggests that apparently costly mechanisms of behavioural thermoregulation can greatly increase fitness if the optimal body temperature is achieved. Field studies of ectotherm thermoregulatory strategies under extreme cold conditions and experiments testing the effects of cool environments on thermoregulatory behaviour are surprisingly scarce. We conducted laboratory experiments to test if common lizards Zootoca vivipara (an active thermoregulator in the field) are able to switch between active thermoregulation and thermoconformity in response to different thermal environments. We found that lizards in treatments with an opportunity to reach their preferred body temperature thermoregulated accurately, maintained their level of daily activity and improved their body condition considerably. In contrast, lizards in the treatment where the preferred body temperature could not be reached became thermoconformers, decreased their daily activity (except for gravid females) and did not increase their body condition. Our results show that lizards can indeed change their thermoregulatory strategy but stress that maintaining the preferred body temperature and, thus, optimising the physiological performance have high priority in lizard behaviour.     

Kin discriminators in the eusocial sweat bee <Emphasis Type="Italic">Lasioglossum malachurum</Emphasis>: the reliability of cuticular and Dufour’s gland odours

The ability to discriminate degrees of relatedness may be expected to evolve if it allows unreciprocated altruism to be preferentially directed towards kin (Hamilton in J Theor Biol 7:1–16, 1964). We explored the possibility of kin recognition in the primitively eusocial halictid bee Lasioglossum malachurum by investigating the reliability of worker odour cues that can be perceived by workers to act as indicators of either nest membership or kinship. Cuticular and Dufour’s gland compounds varied significantly among colonies of L. malachurum, providing the potential for nestmate discrimination. A significant, though weak, negative correlation between chemical distance and genetic relatedness (r = −0.055, p < 0.001) suggests a genetic component to variation in cuticular bouquet, but odour cues were not informative enough to discriminate between different degrees of relatedness within nests. This pattern of variation was similar for Dufour’s gland bouquets. The presence of unrelated individuals within nests that are not chemically different from their nestmates suggests that the discrimination system of L. malachurum is prone to acceptance errors. Compounds produced by colony members are likely combined to generate a gestalt colony chemical signature such that all nestmates have a similar smell. The correlation between odour cues and nest membership was greater for perceived compounds than for non-perceived compounds, suggesting that variability in perceived compounds is a result of positive selection for nestmate recognition despite potentially stabilising selection to reduce variability in odour differences and thereby to reduce costs derived from excessive intracolony nepotistic behaviour.     

Preservation and loss of the honey bee (<Emphasis Type="Italic">Apis</Emphasis>) egg-marking signal across evolutionary time

Honey bee workers are able to distinguish queen-laid eggs from worker-laid eggs, and remove (‘police’) worker-laid eggs. The cue that police workers use is as yet unidentified but is likely to be a chemical signal. This signal benefits queens for it ensures their reproductive monopoly. It also benefits collective workers because it allows them to raise more closely related queen-laid males than the less-related sons of half sisters. Because both parties benefit from the egg-marking signal, it should be stable over evolutionary time. We show that Apis mellifera workers can distinguish queen-laid from worker-laid eggs of the dwarf honey bee A. florea, a phylogenetically distant species that diverged from the A. mellifera lineage 6–10 mya. However, A. mellifera workers are unable to distinguish worker-laid eggs of A. cerana, a much more recent divergence (2–3 mya). The apparent change in the egg-marking signal used by A. cerana may be associated with the high rates of ovary activation in this species.     

Exposing males to female scent increases the cost of controlling <Emphasis Type="Italic">Salmonella</Emphasis> infection in wild house mice

Secondary sexual characters often provide indicators of a male’s resistance to infectious diseases to rivals and potential mates, but it is unclear why. It is often suggested that males honestly signal their health due to energetic and other physiological trade-offs between investing into secondary sexual traits vs resistance to infectious diseases. Our aim was to determine whether such a trade-off exists using wild-derived male house mice (Mus domesticus). We exposed male mice to female scent, a manipulation that induces elevations in testosterone concentration and the expression of a variety of testosterone-mediated secondary sexual traits, and tested whether this sexual stimulation impaired the males’ ability to resolve or cope with an experimental infection (Salmonella enterica). We kept the males on a controlled diet to prevent them from compensating by eating more food. We found that sexually stimulated males were able to control bacterial growth as effectively as sham-stimulated controls; however, to do so, they lost more body mass during infection compared to the controls. In contrast, we found no evidence that sexual stimulation reduced the body mass of uninfected male mice. These results indicate that males’ responses to female odor are not immunosuppressive per se, yet they increase the energetic costs of controlling infection. Our findings support the idea that there is a physiological trade-off between secondary sexual signaling vs resistance to infectious diseases and suggest that studies using only immunocompetence assays might fail to detect such energetic trade-offs. We dedicate this paper to the late Professor Chris Barnard who conducted pioneering research on this topic.     

Regulation of body dimethylsulfoniopropionate (DMSP) content by the copepod Temora longicornis: a test of four mechanisms

Copepods contain dimethylsulfoniopropionate (DMSP) in their tissues in addition to the DMSP in their gut contents and therefore constitute an additional pool of DMSP in the ocean. In the estuarine copepod Temora longicornis Müller, this dynamic pool of DMSP is reduced when external salinity decreases. In the present study the mechanism(s) used by T. longicornis to adjust its DMSP content were examined. Four possible mechanisms were tested in experiments conducted between July 1997 and May 1999: (1) DMSP cleavage, (2) demethylation, (3) conversion to methionine, and (4) release from the body. Tissue extracts of T. longicornis did not exhibit the ability to cleave or demethylate DMSP. In incubation experiments, when external salinity decreased, T. longicornis individuals reduced their DMSP content without increasing their methionine content. The total amount of DMSP in the incubation vials was conserved regardless of salinity. The copepods retained most of the DMSP in their tissues in 30‰ water, but when salinity was reduced to 20‰, the copepods released 41 to 55% of their body DMSP to the surrounding medium; 89 to 91% of the total DMSP was recovered. This suggests that estuarine copepods represent a reservoir of DMSP, which is released as dissolved DMSP upon salinity decline. Therefore, while osmoregulation by these copepods may have little impact on the chemical properties of DMSP, it will affect the partitioning of DMSP between particulate and dissolved phases in the water column. Received: 20 August 1999 / Accepted: 28 March 2000     

Embryonic,larval, and post-metamorphic development of the sea urchin <Emphasis Type="Italic">Cassidulus mitis</Emphasis> (Echinoidea; Cassiduloida): an endemic brooding species from Rio de Janeiro,Brazil

Cassiduloids are currently rare irregular echinoids with a highly conserved adult morphology. Aristotle’s lantern is present only during the post-metamorphic stage, and little is known about the early development of species in this group. Cassidulus mitis produces eggs of about 375 μm in diameter, lecithotrophic larvae with four reduced arms with skeletal fenestrated rods, cilia along the body surface, and a ciliated band on arms and lobes. Offspring is brooded among the female spines from embryo to settler’s stage. The echinopluteus larval stage is reached 6 days after fertilization, and the settler’s stage is formed at the age of 17 days. Aristotle’s lantern appears around the thirteenth day of development. The lantern is well developed and functional in settlers. It remains until at least 62 days after fertilization and can be used to acquire food from the environment. The early development of C. mitis is unusual concerning features of typical lecithotrophic larvae (such as reduced arms), but retains some features of planktotrophic larvae (such as skeletal rods and a ciliated band). Regarding egg size, early development in C. mitis seems to be transitioning from facultative lecithotrophic to typical obligate lecithotrophic pattern in echinoid larval evolution.     

Functional morphology and phylogeny of the rock-boring bivalves <Emphasis Type="Italic">Leiosolenus</Emphasis> and <Emphasis Type="Italic">Lithophaga</Emphasis> (Bivalvia: Mytilidae): a third functional clade

The functional morphology of the shell of rock-boring mytilids (especially Leiosolenus and Lithophaga) is analyzed and compared with that of several epifaunal and semi-infaunal mytilids. Semi-infaunal species are generally intermediate between epifaunal and rock-boring ones both in terms of shell form and the magnitude of forces pulling the shells against the substratum. A molecular phylogenetic analyses using 18s rDNA sequence data strongly suggests that Leiosolenus and Lithophaga are monophyletic genera but that the so-called Lithophaginae (or Leiosolenus plus Lithophaga) is a paraphyletic group. The common cylindrical shell form of rock-boring species, which is here called “lithophagiform” as a third functional mytilid clade, may be due to convergence, as is the case with ‘mytiliform’ and ‘modioliform’.     

Stingless bee response to spider webs is dependent on the context of encounter

In the course of their foraging bouts, bees frequently encounter spider webs among the vegetation. The ability to see and avoid these webs is vital for the success of the individual bee’s foraging bout. In this study, we report on the response of stingless bees (Trigona carbonaria) towards the webs of the St. Andrew’s Cross spider (Argiope keyserlingi). We studied the ability of bees to avoid webs in different contexts: when bees were on their foraging path or when they were returning to the hive as well as when they were flying North or South. We show that the probability of a bee being able to avoid a web depends on the context of the bee’s flight rather than the visual appearance of the web. Furthermore, the presence of the spider seems to alert the bee to the web, resulting in bees being more able to avoid capture. We show, specifically, that the probability of being captured is higher when the bee is returning to the hive compared with when the bee is foraging. The likelihood of avoiding a web is also influenced by the compass direction of the flight, although to a lesser extent. Our results indicate that the context of the predator–prey encounter has a significant influence on a bee’s ability to escape interception by a spider web.     

Sex pheromone of the large aspen tortrix, Choristoneura conflictana(Lepidoptera: Tortricidae)

Summary. Three components that elicited antennal response from male Choristoneura conflictana were found from female gland extracts analyzed using a coupled gas chromatographic-electroantennographic detector system. The main component in gland extracts was (Z)-11-tetradecenal (Z11-14:Ald). Two minor components also elicited antennal response: (E)11-tetradecenal (E11-14:Ald) and (Z)-11- tetradecen-1-ol (Z11-14:OH). Analysis of effluvia indicated that calling virgin females release mostly Z11-14:Ald and trace amounts of Z11-14:OH. Field and wind tunnel behavioral studies showed that Z11-14:Ald alone attracted male moths in a dose response pattern. Tests comparing male response to blends of components detected in gland extracts showed that addition of 1.8% of E11-14:Ald to Z11-14:Ald did not influence male moths in the wind tunnel, but resulted in significantly lower trap captures in the field. The threecomponent blend [Z11-14:Ald (100), E11-14:Ald (1.8), Z11-14:OH (11)], was less attractive than Z11-14:Ald alone in both field and wind tunnel studies. Traps baited with two virgin female moths were equally attractive to males as traps baited with the three-component synthetic blend but less attractive than traps baited with Z11-14:Ald alone. Field tests of various blends of the two components (Z11-14:Ald, Z11-14:OH) detected in the females’ effluvia showed that the addition of 1–10% Z11-14:OH to Z11-14:Ald did not affect the males’ response to Z11-14:Ald. Our data demonstrate that female C. conflictana release sex pheromone components in a different ratio than they are stored in the pheromone gland. The sex pheromone is comprised of a single component, Z11-14:Ald, that can be used to monitor mated and virgin male C. conflictana throughout their flight period.