Influence of Illuminance and Forager Experience on Use of Orientation Cues in Social Wasps (Vespinae)

We tested the influence of illuminance and level of forager experience on nest orientation behavior of the social wasps Vespula vulgaris, Vespa crabro, and Dolichovespula saxonica in an artificial laboratory tunnel system. The number of wasps which oriented themselves chemically via a terrestrial trail or used visual orientation were determined at different illuminance levels for foragers which were naïve or experienced with the tunnel system. In V. vulgaris and D. saxonica, mainly the young and naïve foragers used the chemical trail for orientation in brightness. Experienced foragers used visual cues for nest orientation. In V. crabro, naïve and experienced foragers followed the chemical trail in a similar intensity. In darkness, when visual orientation was limited, the relative importance of the chemical trail increased dramatically in all species and all experience classes.     

Chemical and behavioural studies of the trail-following pheromone in the leaf-cutting ant Atta opaciceps,Borgmeier (Hymenoptera: Formicidae)

To understand the significance of the trail pheromone used in chemical communication of the leaf-cutting ants Atta opaciceps we investigated, under laboratory conditions, the trail-following behaviour of different castes. We observed a clear behavioural discrimination of conspecific venom gland extract of foraging ants from those of other species. Additionally, we determined the pheromone composition of A. opaciceps venom gland secretion using a two-dimensional gas chromatography coupled with mass spectrometry. Chemical analyses revealed the presence of three nitrogen-containing compounds, identified as 2,5-dimethylpyrazine, 3-ethyl-2,5-dimethylpyrazine and methyl 4-methylpyrrole-2-carboxylate (M4MPC). Four different bioassays performed with workers from different castes of A. opaciceps suggested that the trail pheromone elicits the trail-following behaviour in conspecifics of all castes, but the foragers respond more strongly to their own pheromone than to that of other castes (gardeners, generalists and soldiers). In addition, A. opaciceps foragers follow the trails made with the venom gland extracts of the unrelated Acromyrmex subterraneus subterraneus foragers as well as they follow the trails made with their own venom gland extract. M4MPC was identified to be the most abundant and the most behaviourally active component of the venom gland extract of A. opaciceps foragers.     

Task-Related Variation of Cuticular Hydrocarbon Profiles Affect Nestmate Recognition in the Giant ant Dinoponera quadriceps

Recognition seems to be one of the more remarkable characteristics of social groups. In social insects, cuticular hydrocarbons are important for colonial recognition by providing a chemical signature for colony members. The acceptance threshold model predicts that colony members will accept conspecifics when the levels of nest mate cues dissimilarities are below the acceptance threshold. We tested the hypothesis that the encounter of a guard ant worker with a nurse may cause a delay in the process of recognition, because nurses from distinct colonies may share greater amount of chemical compounds. Dinoponera quadriceps guard workers decreased their effectiveness to recognize nurses rather than did to foragers. Alien foragers received significantly more bites and other stronger acts than non-nestmate nurses when they were experimentally introduced. In addition, guards took significantly more time to react against non-nestmate nurses than against alien foragers. Analysis of the cuticular hydrocarbon profiles corroborated our behavioural analysis that nurses from distinct colonies overlap greater amount of cuticular hydrocarbons.     

Interactions with Combined Chemical Cues Inform Harvester Ant Foragers' Decisions to Leave the Nest in Search of Food

Social insect colonies operate without central control or any global assessment of what needs to be done by workers. Colony organization arises from the responses of individuals to local cues. Red harvester ants (Pogonomyrmex barbatus) regulate foraging using interactions between returning and outgoing foragers. The rate at which foragers return with seeds, a measure of food availability, sets the rate at which outgoing foragers leave the nest on foraging trips. We used mimics to test whether outgoing foragers inside the nest respond to the odor of food, oleic acid, the odor of the forager itself, cuticular hydrocarbons, or a combination of both with increased foraging activity. We compared foraging activity, the rate at which foragers passed a line on a trail, before and after the addition of mimics. The combination of both odors, those of food and of foragers, is required to stimulate foraging. The addition of blank mimics, mimics coated with food odor alone, or mimics coated with forager odor alone did not increase foraging activity. We compared the rates at which foragers inside the nest interacted with other ants, blank mimics, and mimics coated with a combination of food and forager odor. Foragers inside the nest interacted more with mimics coated with combined forager/seed odors than with blank mimics, and these interactions had the same effect as those with other foragers. Outgoing foragers inside the nest entrance are stimulated to leave the nest in search of food by interacting with foragers returning with seeds. By using the combined odors of forager cuticular hydrocarbons and of seeds, the colony captures precise information, on the timescale of seconds, about the current availability of food.     

Task-related variation of postpharyngeal and cuticular hydrocarbon compositions in the ant Myrmicaria eumenoides

In the ant Myrmicaria eumenoides we investigated postpharyngeal and cuticular hydrocarbons. At eclosion the glands contained almost no hydrocarbons and there were no lipid inclusions in the glandular epithelium. During the first 3 weeks of adult life the amount of hydrocarbons in the gland increased until day 5, and then remained constant while the lipid content in the epithelium increased steadily. Intracolonial hydrocarbon compositions were not uniform. Compositions of post-pharyngeal and cuticular hydrocarbons in individual ants varied simultaneously, but in different manner depending on the tasks of the ant (brood-tenders, foragers, scouts). Variations on the cuticle were greater than in the gland, but they were strongly correlated. Independent of ants' age and task, cuticular hydrocarbon compositions were dominated by alkenes and alkadienes. Task-specific differences in cuticular compositions were mainly in the amount of alkenes (high in foragers) and alkadienes (high in brood-tenders). Variation of hydrocarbons was low in ants up to 10 weeks old. Thereafter, ants fell into two groups: (1) ants that did not change their hydrocarbons and remained in the nest, and (2) ants that changed their hydrocarbon compositions and became foragers. These results contribute to an ongoing discussion of the dynamic relationship between post-pharyngeal and cuticular hydrocarbons.     

Sex, age and ovarian activity affect cuticular hydrocarbons in Diacamma ceylonense, a queenless ant

In the queenless ant, Diacamma ceylonense, the cuticular hydrocarbons (C25-C35) of nestmate workers vary in their proportions according to age and fertility. Newly eclosed adults ('callows') initially have the same cuticular profile, but with time this changes to that typical of foragers. In contrast, workers that begin to produce eggs develop a different cuticular profile. Several substances (n-C29 and some methyl C25 and C27) discriminate these different social categories (callows, foragers and egg-layers). In Diacamma ceylonense, inter-colony variation of the cuticular hydrocarbons was much lower than intra-colony variation. We also found qualitative differences between the sexes, with males having a clearly different profile with much more alkanes. We discuss these results in the context of physiological models of the relation between ovarian activity and the synthesis of cuticular hydrocarbons. Variations in cuticular profile are a reliable reflection of ovarian activity, and could be used by ants as a fertility signal.     

How patrollers set foraging direction in harvester ants

Recruitment to food or nest sites is well known in ants; the recruiting ants lay a chemical trail that other ants follow to the target site, or they walk with other ants to the target site. Here we report that a different process determines foraging direction in the harvester ant Pogonomyrmex barbatus. Each day, the colony chooses from among up to eight distinct foraging trails; colonies use different trails on different days. Here we show that the patrollers regulate the direction taken by foragers each day by depositing Dufour's secretions onto a sector of the nest mound about 20 cm long and leading to the beginning of a foraging trail. The patrollers do not recruit foragers all the way to food sources, which may be up to 20 m away. Fewer foragers traveled along a trail if patrollers had no access to the sector of the nest mound leading to that trail. Adding Dufour's gland extract to patroller-free sectors of the nest mound rescued foraging in that direction, while poison gland extract did not. We also found that in the absence of patrollers, most foragers used the direction they had used on the previous day. Thus, the colony's 30-50 patrollers act as gatekeepers for thousands of foragers and choose a foraging direction, but they do not recruit and lead foragers all the way to a food source.     

Cuticular extracts inducing aggregation in the German cockroach,Blattella germanica (L.)

German cockroaches Blattella germanica (L.) are gregarious insects. An aggregation pheromone contributes to the maintenance of aggregates. Choice experiments checked the efficiency of different solvents, i.e. dichloromethane, methanol and pentane, in extracting attractive substances and compared the attractiveness of extracts of different parts of the body. Dichloromethane and pentane were the most efficient solvents tested for extracting the attractive substances. Methanol whole body extracts appeared inefficient to induce aggregation. The proportion of larvae attracted to conditioned papers decreased in relation to the size of cuticular surface washed, from whole body to half-body and again to a section of the body cut in three. Attractive substances appear to be present on all parts of the body. Chemical analysis by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) showed that the active extracts contained only cuticular hydrocarbons. In addition to behavioural tests, differences between the composition of methanol extracts and that of the extracts for the other two solvents were revealed by GC. These results indicated that the cuticular hydrocarbons operate as an aggregation pheromone in Blattella germanica.     

The role of cuticular hydrocarbons in mating and conspecific recognition in the closely related longicorn beetles Pidonia grallatrix and P. takechii

The role of cuticular chemicals in mating behavior and their chemical components were studied in two sympatric flower-visiting longicorn beetles, Pidonia grallatrix and P. takechii. Mating experiments revealed that female cuticular chemicals elicit male mating behavior and that males can discriminate between conspecific and heterospecific females on the basis of contact chemicals. GC-MS analyses of whole-body extracts in the two species and both sexes determined that extracts contained a series of hydrocarbons including n-alkanes, n-alkenes, and methylalkanes. The relative abundance of some hydrocarbons differed between species and sexes, and canonical discriminant analysis showed discrimination of species and sex could be made unambiguously with several compounds. These results imply that the difference in cuticular hydrocarbons facilitates the premating isolation of sympatric Pidonia species.     

Correlation between mandibular gland secretion and cuticular hydrocarbons in the stingless bee Melipona quadrifasciata

We investigated whether Melipona quadrifasciata worker mandibular gland secretions contribute directly to their cuticular hydrocarbon profile. The mandibular gland secretion composition and cuticular surface compounds of newly emerged worker bees, nurse bees, and foragers were determined by gas chromatography and mass spectrometry and compared. Both the mandibular gland secretions and the cuticular surface compounds of all worker stages were found to be composed almost exclusively of hydrocarbons. Although the relative proportion of hydrocarbons from the cuticular surface and gland secretion was statistically different, there was a high similarity in the qualitative composition between these structures in all groups of bees.     

The influence of mechanical, visual and contact chemical stimulation on the behavioural phase state of solitarious desert locusts (Schistocerca gregaria)

We investigated the influence of mechanical, visual and contact chemical stimulation on behavioural gregarization of fifth-instar solitarious nymphs of the desert locust. The stimuli were applied in two 2x2 factorial experimental designs, the first with contact chemical and mechanical stimuli, and the second with contact chemical and visual stimulation. Stimulus treatments were applied for a 4-5 h period, after which the behavioural phase state of individual locusts was measured using an assay based on multiple logistic regression analysis of behavioural variables. Mechanical stimulation was provided by showering test insects with millet seeds, thereby excluding the possibility of contact chemical self-stimulation by repeated contact with the same objects. Visual stimulation consisted of the sight of crowd-reared locusts, while contact chemical stimulation was a dichloromethane extract of cuticular hydrocarbons from gregarious nymphs applied to the perch of the test insect. Mechanical stimulation was powerfully gregarizing, whether alone or in combination with contact chemical stimuli. Application of cuticular extract to the perch had no measurable effect on behavioural phase state, either alone or when presented with mechanical or visual stimuli. Visual stimulation alone partly gregarized test locusts. These results appear to conflict with other reports of the gregarizing effect of cuticular hydrocarbons and possible reasons for this discrepancy are discussed.     

Behavioral Activity of Hydrocarbons Emitted by Honeybee Waggle Dancers

Waggle-dancing honeybee foragers emit four hydrocarbons that have been shown to stimulate colony foraging by reactivating experienced foragers and increasing the number of recruitment dances. These hydrocarbons, the alkanes tricosane and pentacosane, and the alkenes (Z)-9-tricosene and (Z)-9-pentacosene, are part of the array of social insects cuticular lipids which have been well studied in the context of nestmate recognition. This study seeks to determine which of the dance hydrocarbons produce the behavioral responses of forager bees by using a binary choice behavioral assay for attraction or repulsion to the volatile phase of the dance compounds. We found no significant deviation from random choice for single dance compounds and pairs of compounds, but bees were significantly attracted to a mixture of the three compounds (Z)-9-tricosene, tricosane, and pentacosane. These results suggest synergy among the waggle-dance hydrocarbons, which was unexpected based on previous research where, in the context of nestmate recognition, alkenes have been shown to play a key role in eliciting behavioral responses. Synergy among the waggle-dance hydrocarbons may provide specificity that facilitates adaptive, context-specific behavioral responses to this subset of cuticular hydrocarbons.     

Cuticular hydrocarbons of Heterotermes tenuis (Isoptera: Rhinotermitidae): analyses and electrophysiological studies

Termites have become an important pest of Eucalyptus and Pinus reforestations, sugarcane and other cultures. An alternative for the control of this pest would be the use of attractive traps that take in account the social behavior of these insects. Diverse factors are important for the insects in the localization of the habitat and the choice of the food and specific odors can facilitate this. Studies referring to Heterotermes tenuis (Isoptera: Rhinotermitidae) are scarce. The objective of this work was to analyze the tergal cuticular extract of H. tenuis and determine the selectivity and sensitivity of its antennae to the components of this extract by electroantennography (EAG). The composition of the cuticular extract was determined by GC-MS analysis. The hydrocarbons found were restricted to linear alkanes, being most abundant C24 to C27 that comprises ca. 65% of the total. Olefins were not detected. EAG and behavioral test responses to the cuticular hydrocarbons were greater and significantly different from the control and the high selectivity of the antennae to the extract indicates its potential as chemical messenger. Cuticular hydrocarbons mixture is species-specific and can be used to identify a given taxon without the diagnostic castes, soldiers or imagoes Difference in the composition appears to relate with the type of habitat of specie.     

白蚁采食行为中的信息交流

白蚁作为社会性昆虫, 其采食个体间依赖各种信息交流机制进行联系与协作, 其中包括踪迹、 警戒、 食物信息、 同伴识别和助食等。通常的联系信号为挥发性或半挥发性化学物质或者一定频谱和功率的机械振动波。其中踪迹信息素、 蚁源烃类、 警戒信息素、 助食素和机械振动信号等都在白蚁采食过程中起着重要作用。白蚁采食过程中食物定向和食物品质信息的传达主要依靠腹板腺分泌的踪迹信息素, 已发现有十二碳烯醇类、 降碳倍半萜类和大环二萜类。蚁源的机械振动也在食物品质表达方面起到一定作用, 但食物品质的表达机制还不明确。白蚁采食协作的基础是同伴识别, 蚁源烃类（C21~C35）是同伴识别的主要信息物质, 采食个体分泌的助食素则可促成共同取食。警戒信息素传递白蚁采食的安全信息, 通常为一些兵蚁源的萜类物质, 但得到功能鉴定的结构还不多。近来研究还发现特殊的蚁源机械振动也可起到示警作用。已初步证实各种信息交流机制间存在交互作用, 但交互作用的机理有待进一步解析。生物物理因素在白蚁采食行为中的作用值得更多重视。本文以白蚁的采食行为为线索, 评述白蚁采食个体间信息交流机制的研究进展及利用问题, 并对今后的研究提出展望。     

Chemical trail systems,orientation, and territorial interactions in the antLasius neoniger

Foraging and territoriality in the ant Lasius neonigerinvolves a series of trails which channel foragers away from adjacent colonies. Experimental studies suggest that the trails are composed of colony-specific, persistent orientation components of hindgut material that accumulate on trails during foraging. A less durable component of the hindgut trail pheromone regulates recruitment. Foraging directionality and the use of a trail could be modified by experimentally arranging confrontations with conspecifics. The orientation of foragers is mediated by visual as well as chemical cues. Components of the foraging and territorial system of L. neonigerappear to include (1) a network of subnests which change in position seasonally within each polydomous nest; (2) a series of trails emanating from each subnest that adjusts search toward resource patches and away from aggressive, neighboring conspecifics; and (3) trail communication involving an ephemeral component of the hindgut trail pheromone that regulates the organization of cooperative prey retrieval and a more persistent component that serves as an orientation guide.     

Not only cuticular lipids: first evidence of differences between foundresses and their daughters in polar substances in the paper wasp Polistes dominulus

Several studies have shown that differences in the relative abundance of cuticular hydrocarbons occur between reproductives and non-reproductives in many social insects. These differences also exist between Polistes dominulus foundresses and their first emerged daughters (usually indicated as workers), but they gradually disappear when workers from orphaned colonies develop their ovaries and assume a reproductive role in the colony. However, hydrocarbons are not the exclusive components of cuticular layer of wasps. Mass-spectrometry analysis of cuticular methanol extracts from Polistes paper wasps showed a complex pattern of polar substances, partly or totally proteinaceous in nature. We found that these compounds, ranging from 918 to 2679 Da, showed a clear caste differentiation between foundresses and their first emerged daughters (usually indicated as "workers"), both in queen-right and in orphaned colonies. Conversely to hydrocarbons, workers from orphaned colonies maintain a significant difference from foundresses in the pattern of the medium molecular weight (MW) polar compounds obtained by MALDI-TOF. On the basis of such results we hypothesize that a reliable cue to identify foundresses from daughters, and not only their reproductive status, may exist in Polistes wasps. Although the great majority of previous work on social insect communication has focused on cuticular hydrocarbons, our findings suggest that the medium MW component of cuticular substances may be involved in recognition.     

Long-term stability of hornet cuticular hydrocarbons facilitates chemotaxonomy using museum specimens

Cuticular hydrocarbons are key compounds used for insect chemical communication and their species-specificity makes them of great utility to chemotaxonomists. However, very little is known about their long-term stability in relation to their use as reliable taxonomic tools. We compared the cuticular hydrocarbon profiles of fresh extracts from four hornet ( Vespa ) species with extracts from specimens that were frozen for 1 year and of those stored in insect display boxes for 20 years. Cuticular hydrocarbon profiles were qualitatively very stable, maintaining their species-specific profiles even after 20 years. The long-term stability of cuticular hydrocarbons in hymenopterans opens up the possibility of using museum collections for chemotaxonomy studies and helping with the delineation of species in difficult groups.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 732–737.     

The allocation of foragers in red wood ants

Abstract. 1. We studied how colonies of the red wood ant, Formica polyctena , adjust the numbers of foragers allocated to different foraging trails. In a series of field experiments, foragers were marked and transferred from one nest to another, related nest, where they joined the foraging force. Transferred workers acted as a reserve of uncommitted, available foragers.
2. Previous work shows that each individual forager habitually uses one trail. We found that for an uncommitted forager, the influence of recruitment initially is stronger than that of directional fidelity. Transferred workers were likely to use trails leading to new food sources. When transferred to a new nest, foragers were not likely to use a trail in the same direction as their original trail in the donor nest.
3. After a week, transferred foragers tended to develop route fidelity. Even after bait was no longer present, they continued to use the trail that had formerly led to a bait source.
4. We examined how colonies adjust numbers on a trail by experimentally depleting some trails. Colonies usually did not compensate for depletion: foragers were not recruited to depleted trails.
5. In general, the dynamics of foraging in this species facilitate a consistent foraging effort rather than rapid adjustments of forager allocation.     

Effects of a neonicotinoid insecticide (acetamiprid) on acetylcholinesterase activity and cuticular hydrocarbons profil in German cockroaches

Acetamiprid was incorporated into the diet at 2% dose corresponding to the LD50 and orally administrated to newly emerged adults of the German cockroach Blattella germanica and investigated on acetylcholinesterase activity and cuticular hydrocarbons profil. Acetylcholinesterase specific activity was determined on adult males and females after 24, 48 and 72 hours of treatment. Pentanic extracts of cuticular hydrocarbons in males and females after 6 days of treatment were analysed by gas chromatography. Data revealed an increase in acetylcholinesterase activity in both sexes from the control series. However, a significant inhibition in AChE was observed after treatment at 24, 48 and 72 hours especially in females. In addition, hydrocarbons profils were found qualitatively similar in all groups of insects. However, slight quantitative differences between sexes in control series were noted. Acetamiprid feminize the cuticular profil in males with significant reduction of cuticular compound, and these allowed separation of insects into two groups using multivariate analysis.     

Phylogeography of two European Reticulitermes (Isoptera) species: the Iberian refugium

Chemical, i.e. cuticular hydrocarbons, and molecular data were used to probe the phylogeography of Reticulitermes termites collected from various parts of France, Spain and Portugal. Phylogenetic relationships were inferred from sequences of the internal transcribed spacer (ITS2) of nuclear ribosomal RNA genes as well as from two partial mitochondrial DNA segments, the cytochrome oxidase II gene and a sequence combining the tRNA-Leu gene and fragments of the NADH dehydrogenase I and ribosomal 16S genes. Two species, namely, R. grassei and R. banyulensis, were identified based on an analysis of cuticular hydrocarbons and the identification was confirmed by ITS2 haplotyping. However, phylogeny based on the analysis of mitochondrial DNA was not completely in agreement with the conclusions drawn from the chemical and nuclear data. An analysis of 56 R. grassei colonies revealed intraspecific differentiation into two major lineages with distinct geographical ranges. Whereas analysis of cuticular hydrocarbons showed that R. banyulensis was chemically distinct from R. grassei, analysis of mitochondrial DNA showed its close kinship with the R. grassei lineage occurring in southern Spain. This kinship could be explained by their evolution from a common polymorphic ancestor species in this ice age refugium.