Behavioral responses to the alarm pheromone of the ant Camponotus obscuripes (Hymenoptera: Formicidae)

The alarm pheromone of the ant Camponotus obscuripes (Formicinae) was identified and quantified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Comparisons between alarm pheromone components and extracts from the major exocrine gland of this ant species revealed that the sources of its alarm pheromone are Dufour's gland and the poison gland. Most components of Dufour's gland were saturated hydrocarbons. n-Undecane comprised more than 90% of all components and in a single Dufour's gland amounted to 19 microg. n-Decane and n-pentadecane were also included in the Dufour's gland secretion. Only formic acid was detected in the poison gland, in amounts ranging from 0.049 to 0.91 microl. This ant species releases a mixture of these substances, each of which has a different volatility and function. When the ants sensed formic acid, they eluded the source of the odor; however, they aggressively approached odors of n-undecane and n-decane, which are highly volatile. In contrast, n-pentadecane, which has the lowest volatility among the identified compounds, was shown to calm the ants. The volatilities of the alarm pheromone components were closely related to their roles in alarm communication. Highly volatile components vaporized rapidly and spread widely, and induced drastic reactions among the ants. As these components became diluted, the less volatile components calmed the excited ants. How the worker ants utilize this alarm communication system for efficient deployment of their nestmates in colony defense is also discussed herein.     

Revisiting the trail pheromone components of the red imported fire ant,Solenopsis invicta Buren

Ants use species-specific trail pheromones to coordinate their sophisticated foraging behavior. During the past decades, many trail pheromone components with various structures have been identified in ants, including the red imported fire ant, Solenopsis invicta, a notorious invasive species worldwide. Four compounds, Z,E- (ZEF) and E,E-α-farnesene (EEF), Z,E- (ZEHF) and E,E-α-homofarnesene (EEHF), have been reported as components of S. invicta trail pheromone. However, another study reported an analog of α-farnesene, Z,Z,Z-allofarnesene, as a key trail pheromone component. These contrasting results caused some uncertainty about the trail pheromone composition in S. invicta. In this study, we synthesized ZEF and EEF, ZEHF and EEHF, and reanalyzed the chemicals in the Dufour gland extract and in the trail pheromone fraction of S. invicta worker body extract. The reported isomers of farnesene and homofarnesene were detected and showed trail-following activity, with ZEF as the major compound, while no allofarnesene was found, neither in the Dufour gland extract nor in the whole-body extract. Our results confirm ZEF and EEF, ZEHF and EEHF as trail pheromone components of S. invicta.     

Worker size polymorphism and ethological role of sting associated glands in the harvester ant Messor barbarus

Summary: Though harvester ants are closely similar in ecology, species differ in their worker size polymorphism as well as in the glandular source of their trail pheromones and defensive compounds. In the harvester ant Messor barbarus, we find that the recruitment trail pheromone is located in the Dufour gland, while defence-alarm substances are produced in the poison gland. We also investigated how the glandular development and the ethological response to these abdominal glands are related to worker body size. For both glands, M. barbarus workers show monophasic and nonisometric growths with slopes of allometric regression lines lower than 1. The highest trail-following response is elicited by the Dufour gland secretion from media workers, responsible for most foraging activities in M. barbarus. Aggressive behaviour is more frequently observed in the presence of poison gland secretions from medium and large-sized workers. Differences between species and between worker size classes in the ethological role of sting associated glands are discussed in relation to the foraging ecology and defensive characteristics of harvester ants.     

A sophisticated,modular communication contributes to ecological dominance in the invasive ant <Emphasis Type="Italic">Anoplolepis gracilipes</Emphasis>

The ecological success of ants is founded on cooperative behaviour and a well functioning communication. Particularly invasive ants are able to act highly cooperatively, out-compete other species, and become ecologically dominant. Since ant communication is to a large extent chemical, we investigated the pheromone functions involved in foraging and alarm behaviour of the invasive tropical formicine Anoplolepis gracilipes. Our results suggest that long-lasting orientation cues are located in hindguts, while Dufour glands contain short-term attractants that trigger an effective recruitment. Poison gland effects were intermediate between hindgut and Dufour gland in terms of orientation, attraction and longevity. In contrast to the other pheromone sources, mandibular glands have a repellent effect and are most likely involved in alarm behaviour. Taken together, the pheromone glands of A. gracilipes contain functionally distinct signals with considerable differences in persistence. In this respect, its communication is exceptional in formicine ants. A strikingly similar communication system was previously detected in Paratrechina longicornis, another opportunistic and invasive formicine ant. Based on these similarities and the differences compared to non-invasive formicine ants, we discuss the role of chemical signals for the coordination of efficient foraging. We conclude that a sophisticated communication system can contribute significantly to ecological dominance and invasive success, in concert with other well known traits.     

Exocrine glands of the ant Myrmoteras iriodum

This paper describes the morphological characteristics of nine major exocrine glands in workers of the formicine ant Myrmoteras iriodum. The elongate mandibles reveal along their entire length a conspicuous intramandibular gland, which contains both class‐1 and class‐3 secretory cells. The secretory cells of the mandibular glands show a peculiar appearance, with a branched end apparatus, which is unusual for ants. The other major glands (pro‐ and postpharyngeal gland, infrabuccal cavity gland, labial gland, metapleural gland, venom gland and Dufour gland) show common features for formicine ants. The precise function of the glands could not yet be experimentally demonstrated, and to clarify this will depend on the availability of live material of these enigmatic ants in future.     

Identification of the alkaloidal venoms of some Monomorium ants of Saudi Arabia

The major volatile compounds in the poison glands of two Monomorium ant species from Saudi Arabia have been identified. Monomorium niloticum and Monomorium najrane both contain mixtures of alkyl- and alkenyl-pyrrolidines and -pyrrolines in their venom glands but no Dufour gland volatile compounds were detected. Monomorium mayri showed neither Dufour gland compounds nor venom components detectable by gas chromatography.     

Dufour gland secretions of Myrmica rugulosa and Myrmica schencki workers

The chemical composition and behavioural activities of the secretions of the Dufour glands of Myrmica rugulosa and M. schencki have been studied, as part of an extended study on Myrmica ants. Chemically, the Dufour gland of M. rugulosa is filled with a mixture of hydrocarbons dominated by straight chain alkanes and alkenes with 13 to 19 carbon atoms, as found in M. rubra. Significant quantities of (Z,E)-α-farnesene and its homologues, homofarnesene and bishomofarnesene, are also present. In M. schencki, the major compounds present are homofarnesene and bishomofarnesene. In both species, the very volatile portion of the Dufour gland secretion is identical to that analysed in M. rubra. From an ethological point of view, this very volatile part is efficient in attracting workers at a distance (6 to 8 cm) and in decreasing their wandering movements. No specificity was observed when performing cross-tests with Dufour glands freshly isolated from workers of other Myrmica species, but obvious specificities were detected when testing the less volatile part of the Dufour glands' contents, known to be used for marking newly discovered areas.     

Contents of the poison apparatus of some species of Pheidole ants

Four Old World species of Pheidole ants contain different mixtures of farnesene-type hydrocarbons in their poison apparatus, and the mixture is different between the minor and major workers within a species. A bishomofarnesene (C₁₇H₂₈) provides approximately half of the secretion of the Dufour glands of minor workers of Pheidole pallidula. (Z,E)-α-Farnesene constituted 96% of the Dufour secretion of major workers of P. pallidula, but only 20% of that of minors. The Dufour glands of minor workers of Pheidole sinaitica contain a mixture of farnesene homologues with (Z,E)-α-farnesene and the bishomofarnesene also found in P. pallidula predominant. The mixture in major workers was similar but had, in addition, a small amount of (E)-β-farnesene. The Dufour glands of Pheidole teneriffana minors contain chiefly the same bishomofarnesene found in P. pallidula and P. sinaitica while major workers contain (Z,E)-α-farnesene. Pheidole megacephala minor workers contained small amounts of eight farnesenes, while major workers contained essentially no farnesenes. The poison glands of minor workers of P. pallidula contain 3-ethyl-2,5-dimethylpyrazine. No pyrazine compounds were found in the major workers of P. pallidula or the minor workers of P. sinaitica. The poison glands of the major workers of P. sinaitica contained larger amounts of tetra-substituted pyrazines. No pyrazines were found in the poison reservoirs of major or minor workers of P. teneriffana or P. megacephala.     

Morphology and chemistry of Dufour glands in four ectoparasitoids: Cephalonomia tarsalis,C. waterstoni (Hymenoptera: Bethylidae), Anisopteromalus calandrae,and Pteromalus cerealellae (Hymenoptera: Pteromalidae)

The venom apparatus of four hymenopterous parasitoids, including two bethylids, C. tarsalis (Ashmead) and C. waterstoni (Gahan), and two pteromalids, A. calandrae (Howard) and P. cerealellae (Ashmead), were removed and the associated Dufour glands characterized with respect to their external morphology and chemistry. Dufour glands in all four species have a characteristic translucent appearance that apparently results from their lipid content. The stalked Dufour glands of C. tarsalis and C. waterstoni are pear-shaped and have overall lengths of approximately 0.2 and 0.15 mm, respectively. The thin venom glands are bifurcate and insert through a fine duct into the transparent ovoid- to pear-shaped venom reservoir in these bethylids. In A. calandrae and P. cerealellae the Dufour glands are elongated, tubular structures of ca. 0.35 and 0.8 mm in length, respectively, that constrict to a short stalk that empties into the common oviduct. The venom glands in these pteromalids are simple elongated structures that insert into the sac-like venom reservoir through a fine duct. The chemistry of the volatile contents of the Dufour gland in these four species differs considerably. C. tarsalis Dufour glands contain the same hydrocarbon components as found on the cuticle of this species (Ann. Entomol. Soc. Am. 91:101-112 (1998)), and no other chemicals. The Dufour glands of C. waterstoni also contain only hydrocarbons, most of which are the same as the cuticular hydrocarbons (Ann. Entomol. Soc. Am. 85:317-325 (1992)), but in addition the Dufour gland contains ca. 3% of a mixture of 2,17- and 2,19-dimethyl C(23). A. calandrae Dufour gland chemistry is somewhat more complex than that of either of the two bethylids, but like the bethylids, only hydrocarbons are present. The carbon number range is from C(30) to C(39) and consists of a mixture of n-alkanes (C(30)-C(38)); 3-, 5-, 7-, 9-, 11-, 12-, 13-, 14-, 15- and 17-methyl alkanes; 3,7- and 3,11-dimethyl alkanes; 5,9- and 5,17-dimethyl alkanes; 7,11-, 9,13-, 13,17-, 14,18- and 15,19-dimethyl alkanes; 3,7,11- and 3, 9,15-trimethyl alkanes; and 3,7,11,15-tetramethyl alkanes. The cuticular hydrocarbons of this species have not been previously reported, but they are the same as the Dufour gland hydrocarbons. The Dufour glands of P. cerealellae contain both hydrocarbons and two long-chain aldehydes. Most of the hydrocarbons are identical to those found on the cuticle of this species (Ann. Entomol. Soc. Am. 94:152-158 (2001)), but in addition, 5,9-dimethyl C(27), 5,13-, 5,17- and 5,19-dimethyl C(35), 12- and 14-methyl C(36), 12,16- and 13,17-dimethyl C(36), 13-methyl C(37) and 13,17-dimethyl C(37) are present. The two aldehydes detected in glands from P. cerealellae are n-tetracosanal (C(23)CHO) and n-hexacosanal (C(25)CHO).     

Chemotaxonomy of some Cataglyphis ants from Morocco and Burkina Faso

The taxonomy of Cataglyphis ants has many unsolved problems and chemotaxonomy can provide additional insight for their resolution. We describe here the chemical content of the postpharyngeal and Dufour glands of three Cataglyphis ant species: C. viaticus, C. mauritanicus from Morocco and for the first time a Sub-Saharan Cataglyphis, C. sp. (BF) from Burkina Faso. These three species are very distinct chemically with respect to both the postpharyngeal and Dufour glands. Methyl-alkenes, rare in ants, are characteristic of C. sp. (BF) postpharyngeal glands. A comparison with C. bicolor from Tunisia indicated that C. sp. (BF) can be included into the bicolor group with C. viaticus. We suggest that the content of the Dufour gland is a better phylogenetic indicator than the content of the postpharyngeal gland.     

Multiple functions of fire ant Solenopsis invicta mandibular gland products

Alarm pheromones of social insects are best known for their role in the defence and maintenance of colony integrity. Previous studies with the fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) demonstrate that the mandibular glands of workers (sterile females) and male and female sexuals produce an alarm pheromone, 2‐ethyl‐3,6‐dimethylpyrazine. The function of alarm pheromones in worker ants is well understood and divergent from the function of these compounds in the winged sexual forms. The present study quantifies the amount of pyrazine in the mandibular glands from male and female alate sexuals, as well as queens. Pyrazine production in female alates starts in the late pupal stage and increases until they reach mating flight‐ready maturity; however, after mating flight participation, the pyrazine level declines by >50%. Interestingly, mature male alates lose >85% of their mandibular gland pyrazine during mating flight activity. The results of the present study indicate that male and female sexuals use mandibular gland secretions for mating flight initiation and during mating flights. Furthermore, the ontogeny of mandibular gland products (pyrazine as the marker) from newly‐mated queens to mature colony queens shows a more than two‐fold increase in the amount of pyrazine by 6 months after mating. However, this is followed by a decline to trace amounts in mature colony queens (>2 years old), suggesting a function for mandibular gland products during colony development. Multifunctional use of social insect pheromones is well documented and data are reported in the present study suggesting new roles for mandibular gland products in fire ants.     

Coordination of Raiding and Emigration in the Ponerine Army Ant Leptogenys distinguenda (Hymenoptera: Formicidae: Ponerinae): A Signal Analysis

Several glandular sources of trail pheromones have been discovered in army ants in general. Nevertheless, at present the understanding of the highly coordinated behavior of these ants is far from complete. The importance of trail pheromone communication for the coordination of raids and emigrations in the ponerine army ant Leptogenys distinguenda was examined, and its ecological function is discussed. The secretions of at least two glands organize the swarming activities of L. distinguenda. The pygidial gland is the source of an orientation pheromone holding the group of raiding workers together. The same pheromone guides emigrations to new nest sites. In addition, the poison sac contains two further components: one with a weak orientation effect and another which produces strong, but short-term attraction and excitement. The latter component is important in prey recruitment and characterizes raid trails. This highly volatile recruitment pheromone allows the extreme swarm dynamic characteristic of this species. Emigration trails lack the poison gland secretion. Due to their different chemical compositions, the ants are thus able to distinguish between raid and emigration trails. Nest emigration is not induced chemically, but mechanically, by the jerking movements of stimulating workers.     

Gastral exocrine products of a myrmicine ant strongly overlap pygidial gland products of Dolichoderinae

In most subfamilies of ants (Formicidae), a pygidial gland occurs in the worker’s dorsal gaster, and where tested, products of this gland function in alarm/defense and/or recruitment communication. These products are well characterized for members of the subfamily Dolichoderinae but remain unidentified for Myrmicinae. Both major and minor workers of one myrmicine, Pheidole biconstricta Mayr, are known to have greatly hypertrophied pygidial glands with alarm/repellent products. GC/MS analysis of methanol extracts of worker gasters revealed actinidine and six diastereomers of iridodials as major components, as well as lesser amounts of two lactones, iridomyrmecin and dihydronepetalactone. The iridoids also occur in the pygidial glands of some dolichoderines. Results are related to functional and phylogenetic considerations. Received 20 December 2004; revised 25 February 2005; accepted 4 March 2005. Work performed at Estación Biológica Cocha Cashu, Madre de Dios, Peru, and Virginia Military Institute, Lexington, VA.     

Effect of gland extracts on digging and residing preferences of red imported fire ant workers (Hymenoptera: Formicidae)

There is evidence that ant‐derived chemical stimuli are involved in regulating the digging behavior in Solenopsis invicta Buren. However, the source gland(s) and chemistry of such stimuli have never been revealed. In this study, extracts of mandibular, Dufour's, postpharyngeal, and poison glands were evaluated for their effect on ant digging and residing preferences of S. invicta workers from three colonies. In the intracolonial bioassays, workers showed significant digging preferences to mandibular gland extracts in 2 of 3 colonies and significant residing preferences in 1 of 3 colonies; significant digging preferences to Dufour's gland extracts in 1 of 3 colonies and significant residing preferences in 2 of 3 colonies. No digging and residing preferences were found for postpharyngeal and poison gland extracts. In intercolonial bioassays, significant digging and residing preferences were found for mandibular gland extracts in 3 of 6 colony combinations. Significant digging preferences to Dufour's gland extracts were found in 4 of 6 colony combinations and significant residing preferences in all 6 colony combinations. For postpharyngeal gland extracts, significant digging preferences were found only in 1 of 6 colonial combinations and no significant residing preferences were found. For poison gland extracts, no significant digging preferences were found; significant residing preferences were found in 1 of 6 colony combinations. However, a significant residing deterrence (negative residing preference index) was found for 2 of 6 colony combinations. Statistical analyses using data pooled from all colonies showed that mandibular and Dufour's gland extracts caused significant digging and residing preferences in both intracolonial and intercolonial bioassays but not postpharyngeal and poison gland extracts. By analyzing the data pooled from the same three colonies used for gland extract bioassays, it was found that, in no cases, workers showed significant digging and residing preferences to 2‐ethyl‐3,6‐dimethylpyrazine, an alarm pheromone component from mandibular gland.     

Biosynthesis of formic acid by the poison glands of formicine ants

The biosynthesis of formic acid in the poison glands of formicine ants is closely related to the C-1 metabolism of the glandular cells. Experiments utilizing radiolabeled amino acids revealed that serine is a major precursor, contributing both its α and β carbons to formic acids. 5,10[¹⁴C]methylene H₄folate and 5,10[¹⁴C]methenyl H₄folate also serve as precursors of formic acid in the poison gland, suggesting that they are intermediates in the pathway. Furthermore, these H₄folate derivatives were isolated from poison glands following incubation with [3-¹⁴C]serine and proved radioactive. The glandular cells are also exceptionally rich in the enzymes responsible for these reactions, supporting the proposed pathway.Although this pathway has been established in various organism, the uniqueness of the poison gland system is that it accumulates formic acid to large extent, yet avoids its cytotoxicity. This is made possible by a combination of the biochemical characteristics of the pathway and the special morphological features of the poison gland.     

Formic acid and saturated hydrocarbons as alarm pheromones for the ant Formica rufa

Releasers for the most intense, the fast-running, phase of the alarm behaviour were studied in worker ants of the formicine ant Formica rufa. The investigation was performed with a new technique in which the intensity and the duration of the behaviour pattern were measured in an objective and automatic way.Workers of formicine ants eiect a mixture of formic acid and the secretion from Dufour's gland against an enemy. The secretion from this gland in F. rufa consists of a great number of substances, 39 of which have been identified. The dominating substances form a homologous series of aliphatic, saturated hydrocarbons. Some of these hydrocarbons as well as the formic acid are alarm pheromones. The behavioural threshold value for one of these compounds, decane, was lower than 5.10¹³ molecules cm^?3 air. The threshold value for formic acid was estimated to 7.10¹⁵ molecules cm^?3 air. Formic acid and undecane are shown to belong to different reaction groups. The hydrocarbons, on the other hand, seem to affect the same kind of acceptor or receptor.The total intensity and duration of the alarm behaviour released by formic acid and one hydrocarbon are additive if the two substances are combined in a stimulus. A combination of two hydrocarbons and formic acid release a stronger behaviour than one hydrocarbon with formic acid even when the two stimuli contain the same number of molecules. The hydrocarbons have a combined effect and their relative concentrations regulate the intensity and duration of the alarm behaviour.     

Behavioral and olfactory antennal responses of Solenopsis geminata (Fabricius) (Hymenoptera: Formicidae) workers to their dufour gland secretion

Behavioral and electrophysiological tests were performed to evaluate the responses of workers of the ant Solenopsis geminata (Fabricius) from different size categories to Dufour gland extracts. Morphometric measures based in head widths across eyes were used to determine worker sizes. Trail following response of different worker sizes to Dufour gland extract from workers of different sizes was assessed. For each worker size category olfactory responses to Dufour gland extracts were determined using electroantennography (EAG). Gas chromatography and mass spectrometry (GC-MS) were used to determine the chromatographic profile of Dufour gland secretion for each worker size. Morphometric measures permitted to classify the workers of S. geminata as large, medium and small workers. Medium S. geminata workers displayed a significantly higher behavioral response to Dufour gland extracts produced by medium size workers. Similarly, medium workers showed a significantly higher EAG response to Dufour gland extracts produced by medium sized workers. Chromatographic profile of Dufour gland secretions produced by workers showed that each size category exhibited a characteristic profile of the three main components considered as potential trail pheromone constituents. This work showed that medium workers of S. geminata exhibited a high trail-following behavior as well as a high antennal response to Dufour gland secretion. This and their relative abundance in field foraging areas, suggest that medium-sized workers are specialized in foraging activities.     

Social Insect Pheromones: Their Chemistry and Function

Exocrine secretions of social insects are often characterizedby extraordinarily complex mixtures of natural products. Thus,chemical communication in social insects must be interpretedin terms of signals generated by multicomponent systems, theindividual constituents of which can affect the informationalcontent of the message. Alarm pheromones have been identified chiefly in three subfamiliesof ants and their distribution appears to be chemosystematicallysignificant. Myrmicine genera emphasize 3-alkanones as alarmreleasers, whereas methyl ketones, primarily of terpenoidalorigin, are widely utilized as alarm pheromones in the subfamilyDolichoderinae. Formicine species may employ formic acidas analarm pheromone in addition to the compounds produced in themandibular and Dufour's glands. The mandibular gland pheromonesare chiefly acyclic monoterpene aldehydes (e.g., citronellal)which are relatively low boiling compounds. Higher boiling n-alkanesare produced in the Dufour's glands and may serve as more persistentreleasers of alarm behavior. Alarm pheromones as well as thecaste-specific pheromones of male bees and ants, probably alsoserve as defensive products. In many cases it is likely thatpheromones were originally utilized as defensive compounds andtheir communicative function is a secondary development.     

Monoenyl hydrocarbons in female body wax of the yellow peach moth as synergists of aldehyde pheromone components

The non-polar components of female body wax and pheromone gland extracts of the yellow peach moth synergistically enhanced male behavioral responses from close to pheromone sources in wind tunnel tests when mixed with an aldehyde pheromone blend. When the non-polar fractions (NPFs) of female body wax were further separated by column chromatography, synergistic activities were found in the 3 and 50% ether in hexane fractions, and they additively increased male responses. The main components of the first fraction were (Z)-9-tricosene, (Z)-9-pentacosene, (Z)-9-heptacosene, (Z)-9-nonacosene and (Z)-9-hentriacontene. Only (Z)-9-heptacosene showed a significant synergistic effect in enhancing male responses, but the other components had no effect. A mixture of the five monoenyl hydrocarbons lost activity at lower doses than 5 ng. Natural ratios of these hydrocarbons in the female body wax and pheromone gland extracts were similar, but the amount of (Z)-9-heptacosene in the female body wax was significantly higher than in the pheromone gland extracts. We conclude that (Z)-9-heptacosene increases male responses to aldehyde pheromones, and unknown component(s) in the 50% ether in the hexane fraction are required for full synergistic enhancement by the NPFs of the female body wax and the pheromone gland extracts.     

Chemical interference competition by Monomorium minimum (Hymenoptera: Formicidae)

Summary Workers of Monomorium minimum forage above-ground for dead arthropods. Small particles (<1 mg) are retrieved individually, but larger particles stimulate recruitment and are dissected by groups of workers. The recruitment pheromone originates in the Dufour's gland and the number of ants responding to a trail varies with pheromone concentration. When ants of other species are encountered at food resources, workers of M. minimum gaster-flag and extrude an irritating poison gland secretion from the sting. This chemical interference delays invasion by competitors and prolongs the period during which the colony can dissect and retrieve pieces of the food resource. M. minimum recruits at higher temperatures than sympatric ant species. The probability of interference at food baits rises from 5% to 100% when they become too large for a single worker to carry. The probability of food resource loss is higher for baits of intermediate weight (x=18.1 mg) than for those of low weight (x=0.1 mg) or high weight (x=403.1 mg).