Slave-making Polyergus Research Articles

Sex Attractant Pheromone of the Japanese Slave-Making Ant, Polyergus samurai.

The objective of our project was to identify the sex attractant pheromone of queens of the slave-making ant Polyergus samurai, which was characterized as a blend of methyl 6-methylsalicylate and methyl 3-ethyl-4-methylpentanoate. Both compounds were identified in volatiles collected from squashed heads of queens, and in field trials, a blend of the two compounds was highly attractive to males. In contrast, males were not attracted to a blend of methyl 6-methylsalicylate with 3-ethyl-4-methylpentanol. Thus, the pheromone blend of this species is analogous to that of species in the Polyergus lucidus complex from eastern North America, rather than that of the Polyergus breviceps complex from western North America and the European species P. rufescens. These results are discussed in the context of speciation within the genus.

Evidence for adaptive brain tissue reduction in obligate social parasites (Polyergus mexicanus) relative to their hosts (Formica fusca)

Brain investment is evolutionarily constrained by high costs of neural tissue. Several ecological factors favour the evolution of increased brain investment; we predict reduced brain region investment will accompany the evolution of organismal or social parasitism when parasites rely on host behaviour and cognition to solve ecological problems. To test this idea we investigated whether brain region investments differed between obligate slave-making Polyergus mexicanus ant workers and their Formica fusca slave workers. Polyergus workers perform little labour for their colonies; enslaved workers of Formica host species forage, excavate nests and tend the brood. We focused on the calyces of the mushroom bodies, central processing brain regions that are larger in social insect workers that perform complex tasks. As predicted we found lower relative investment in mushroom body calyx in P. mexicanus workers than in F. fusca workers; by contrast, enslaved and free F. fusca workers did not differ in mushroom body calyx volume. We then tested whether slave-makers and hosts differed in brain investment among sensory modalities. Polyergus slave-makers employ several unique classes of pheromones during raids, and eye size relative to head size was smaller in P. mexicanus workers than in F. fusca workers. The size of antennal brain tissues relative to visual tissues was greater in Polyergus, both in the peripheral sensory lobes and in the mushroom body calyx, suggesting greater relative investment in antennal processing by slave-makers. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113, 415–422.

Chemical camouflage of the slave-making ant Polyergus samurai queen in the process of the host colony usurpation (Hymenoptera: Formicidae)

Founding queens of the obligatory social parasite ant Polyergus samurai usurp the host ant Formica japonica colony. The aggressive behaviors of F. japonica workers on the parasite queen disappear after the parasite queen kills the resident queen. To determine whether the parasite queen chemically mimics the host ants, we examined the aggressive behavior of F. japonica workers toward glass dummies applied with various extracts of the parasite queen and host workers. The crude extracts and hydrocarbon fraction reproduced the host workers’ behavior to the live ants. The extracts of the post-adoption parasite queen, as well as the nestmate extracts of F. japonica, did not elicit the aggressive behavior, but the extract of the pre-adoption parasite queen triggered attacks by the host workers. The nestmate recognition of host workers did not change, regardless of contact with the parasite. The gas chromatography and gas chromatography–mass spectrometry analyses indicated that the cuticular hydrocarbon (CHC) profile of the parasite queen drastically changed during the process of usurpation. Discriminant analysis showed the successfully usurped P. samurai queen had colony-specific CHC profiles. CHC profiles of the P. samurai queen who killed the host queen were more similar to those of the host queen than the workers, while the P. samurai queen who usurped the queenless colony had a profile similar to those of host workers. These results suggest that the P. samurai queen usually acquires the CHCs from the host queen during the fight, but from host wokers in queenless host colonies.

Chemical communication in mating behaviour of the slave-making ant Polyergus rufescens (Hymenoptera, Formicidae): 3-ethyl-4-methylpentanol as a critical component of the queen sex pheromone

The aim of the research reported here was to determine whether 3-ethyl-4-methylpentanol, a minor but crucial component of the sex pheromone of the North American slave-making ant species Polyergus breviceps, was also a component of the sex pheromone of the European congener Polyergus rufescens. Thus, the contents of mandibular glands of P. rufescens virgin queen were extracted and analysed. The main component of the extracts was methyl 6-methylsalicylate and 3-ethyl-4-methylpentanol was identified as one of several minor components. Further analyses showed that the insects produce mainly the (R)-enantiomer of the alcohol. Males’ responses to various blends of methyl 6-methylsalicylate with the racemate or the pure enantiomers of 3-ethyl-4- methylpentanol were tested in field behavioural bioassays. The data showed that blends of methyl 6- methylsalicylate and 3-ethyl-4-methylpentanol were strongly synergistic, with the most active ratios being biased toward the first component. The addition of other minor components to the binary blend neither increased nor decreased responses by males. Only the (R)-enantiomer of the alcohol was biologically active; its antipode did not inhibit attraction. The results are discussed in terms of the evolution of signals, and are compared with the results previously obtained for the allopatric species Polyergus breviceps.

REPRODUCTIVE CONFLICT IN SOCIAL INSECTS: MALE PRODUCTION BY WORKERS IN A SLAVE-MAKING ANT.

In insect societies, workers cooperate but may also pursue their individual interests, such as laying viable male eggs. The case of obligatory slave-making ants is of particular interest because workers do not engage in maintenance activities and foraging. Therefore, worker egg laying is expected to be less detrimental for colony efficiency than in related, nonparasitic species. Furthermore, as slave-making workers usually do not perform brood care and thus might have little power in manipulating sex allocation, they might be more strongly selected to increase their direct fitness by producing their own sons than workers in nonparasitic species. In this study we investigated worker reproduction in four natural colonies of the slave-making ant Polyergus rufescens, using highly variable microsatellite markers. Our results show that workers produce up to 100% of the males. This study thus presents the first direct evidence of an almost complete takeover of male reproduction by workers in ants.

Behavioral Bioassays Testing the Methyl 6-Methylsalicylate as a Component of the Female Sex Pheromone in the Slave-Making Ant Polyergus rufescens (Hymenoptera, Formicidae)

Behavioral Bioassays Testing the Methyl 6-Methylsalicylate as a Component of the Female Sex Pheromone in the Slave-Making Ant Polyergus rufescens (Hymenoptera, Formicidae)

Morpho-functional comparison of the Dufour gland in the female castes of the Amazon ant Polyergus rufescens (Hymenoptera, Formicidae)

The Dufour gland is crucially involved in main aspects of the parasite habit of the slave-making ant Polyergus rufescens, i.e. slave-raids and host colony usurpation. Workers use chemicals from this gland as recruitment signals during raid organization, while newly-mated queens use its secretion to appease residents during host nest invasion. Here, we report a comparison of the general morphology and fine structure of the Dufour gland in the female castes of P. rufescens: queens, ergatogynes (intermediate forms), and workers. The analysis clearly shows the link between gland structure and its behavioural role in queens and workers. In particular, queens present a hypertrophied gland with an extended lumen and a thin epithelium no more active in secretory function. This is consistent with the fact that usurper queens use the Dufour gland contents only during the short phase of host nest penetration. Contrary to adult queens, the cytoplasmic organization of the Dufour gland epithelium of raiders is typical for a tissue with secretory activity (abundance of mitochondria, free ribosomes, strands of smooth endoplasmic reticulum and a Golgi apparatus). This is consistent with the continuous raiding activity performed by workers throughout their adult life. The biology of ergatogynes is still an enigmatic matter. Their Dufour gland is intermediate in shape and size between that of queens and workers. It presents a fairly thick epithelium with features that are typical of a quite active secretory tissue.

REPRODUCTIVE CONFLICT IN SOCIAL INSECTS: MALE PRODUCTION BY WORKERS IN A SLAVE-MAKING ANT

In insect societies, workers cooperate but may also pursue their individual interests, such as laying viable male eggs. The case of obligatory slave-making ants is of particular interest because workers do not engage in maintenance activities and foraging. Therefore, worker egg laying is expected to be less detrimental for colony efficiency than in related, nonparasitic species. Furthermore, as slave-making workers usually do not perform brood care and thus might have little power in manipulating sex allocation, they might be more strongly selected to increase their direct fitness by producing their own sons than workers in nonparasitic species. In this study we investigated worker reproduction in four natural colonies of the slave-making ant Polyergus rufescens, using highly variable microsatellite markers. Our results show that workers produce up to 100% of the males. This study thus presents the first direct evidence of an almost complete takeover of male reproduction by workers in ants.

Knowing your enemies: seasonal dynamics of host-social parasite recognition.

Despite its evolutionary significance, behavioural flexibility of social response has rarely been investigated in insects. We studied a host-social parasite system: the slave-making ant Polyergus rufescens and its host Formica rufibarbis. Free-living host workers from parasitized and from unparasitized areas were compared in their level of aggression against the parasite and alien conspecifics. We expected that a seasonal change would occur in the acceptance threshold of F. rufibarbis workers from a parasitized area towards the parasite, whereas F. rufibarbis workers from an unparasitized area would not show substantial changes connected with the parasite's peak in activity (raiding and colony-founding season). The results showed a significant adaptive behavioural flexibility of host species workers and are consistent with the acceptance threshold model's (Reeve 1989) prediction that recognition systems are not fixed but context-dependent. In particular, host workers from the unparasitized area were highly aggressive towards the parasite regardless of the season, whereas host workers from the parasitized area significantly increased their aggression towards the parasite during its raiding and colony-founding season. Being able to detect and possibly kill a Polyergus scout searching for host nests can be an effective strategy for a Formica colony to avoid being raided or usurped by a parasite queen.

Sex Pheromone of Queens of the Slave-Making Ant, Polyergus breviceps

The sex attractant pheromone produced in mandibular glands of queens of the slave-making ant Polyergus breviceps has been identified as a blend of methyl 6-methylsalicylate and 3-ethyl-4-methylpentanol. In field trials, each compound alone was completely unattractive to males, whereas blends of the two compounds attracted hundreds of males within a couple of hours.

Mandible associated glands in queens of the slave-making ant Polyergus rufescens (Hymenoptera, Formicidae)

A first description of the structural organization of two exocrine glands associated with the mandibles of queens of the obligatory slave-making ant Polyergus rufescens is reported. The mandibular gland consists of clustered bicellular secretory units connected by means of cuticular ducts to a big reservoir. The reservoir continues in a duct that opens proximally on the mandible cuticle. Intramandibular glands are isolated bicellular secretory units connected to the external pores through a cuticular duct. No reservoir has been observed. In both cases the secretory cells belong to the 3rd class. Functional aspects of the investigated glands are discussed taking also into account for the parasitic habit of this slave-making species.

How raiders of the slave-making ant Polyergus rufescens (Hymenoptera Formicidae) evaluate a target host nest

Raiding activity is a crucial task of the slave-making ant Polyergus rufescens, during which colonies of the host species are attacked in order to pillage the resident brood and obtain a slave work force. The features of the host colony influencing the organization of P. rufescens raids were investigated by laboratory experiments which allow the analysis of behavioural patterns unlikely to be observed in the field. In particular, the mechanism of evaluation of the target nest was studied. By providing P. rufescens the possibility to sack different host nests, we found that a greater number of raiders was recruited when a greater amount of brood was present inside the attacked nest. The hypotheses that raiding activity is performed in accordance with the “optimal foraging theory” and that raiders prefer to sack colonies where the residents are more aggressive are discussed.

Host queen killing by a slave-maker ant queen: when is a host queen worth attacking?

A new colony of the slave-making ant Polyergus breviceps is initiated when a newly mated gyne invades a host nest and kills the resident queen. This process seems to result in chemical camouflage of the invading gyne and allows her to usurp the position of colony reproductive. Young, recently mated Formica gynes, however, are not attacked. To determine whether worker and/or immature presence is the basis for aggression, we placed eggs, larvae, pupae and workers from mature F.gnava queens with newly mated F.gnava queens and observed the responses of introducedP.breviceps queens. Because no newly mated gyne was attacked, we tested newly mated F.gnava queens (1) once they had produced eggs, (2) when the offspring reached the larval, pupal and callow stages of development, and (3) every 2 weeks until aggression ensued. Eventually all F.gnava queens were attacked but only 29 weeks after having mated. Thus, although offspring are the ultimate benefit from attacking an established F.gnava queen,P.breviceps queens detect mature queens using another time-dependent feature that is reliably indicative of reproductive status. The similarity of host queen hydrocarbon profiles, often correlated with reproductive status in other ant species, suggests that other compounds reflect queen fecundity and produce a kairomonal effect, or that another cue signals host queen and colony suitability. Our findings indicate P.breviceps gynes have evolved to respond aggressively to a host gyne cue that appears long after mating, preventing attacks on gynes without the workers necessary for colony founding.Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.

Glandular sources of recruitment, trail, and propaganda semiochemicals in the slave-making ant Polyergus rufescens

The chemical sources involved in raiding activities of the slave-making ant Polyergus rufescens were investigated in the laboratory. In particular, we identified the sources of substances the raiding workers employ to convey information to nestmates and to disrupt reactions by defending workers during host nest invasion. Different secretions were offered to the dulotic ants on the tip of a blotting paper strip inserted a few centimetres into the nest entrance. The Dufour's gland secretion induced the characteristic mass exit from the nest and is likely the source of a recruitment signal. Moreover, laboratory tests with secretions from different anatomical structures revealed that the trail signals for homing raiding workers are conveyed by their hindgut contents released during the outbound trip. Finally, to ascertain if P. rufescens raiding workers use “propaganda” pheromones, the behavioural response of the Formica (Serviformica) cunicularia host workers to different secretions of the slave-makers was investigated. Only mandibular secretions caused the typical panic reactions shown by residents at the moment of nest invasion. This probably facilitates both nest invasion and brood sacking by raiding workers.

Dufour’s gland contents of queens of the slave-making ant Polyergus rufescens and its host species Formica cunicularia

Dufour’s gland contents of the queens of the slave-making ant species Polyergus rufescens and its host, Formica cunicularia, were analysed by gas chromatography-mass spectrometry. In P. rufescens, the gland contains a mixture of several esters, one of which, decyl isobutyrate, was consistently present in large quantities (constituting more than 80% of glandular contents). Glandular contents in the host species, were found to be different, containing, in addition to two substances also found in P. rufescens, a mixture of acetates and several hydrocarbons. Decyl isobutyrate was not present. We discuss the possible role of these substances in the life history of P. rufescens.

Colony founding in Polyergus rufescens : the role of the Dufour's gland

In the European slave-making ant Polyergus rufescens, the occurrence of chemical strategies during the initial phase of dependent colony foundation or usurpation was investigated. To test this idea, we analysed the effect of the secretion of different glands (Dufour's, poison, pygidial, rectal, and mandibular) on the behaviour of workers of its common host species, Formica cunicularia (subgenus Serviformica). Workers of another species, Formica rufibarbis (Serviformica), were daubed with these extracts, and introduced into colony fragments of F. cunicularia. The results of a set of laboratory aggression test showed that the secretion of the mandibular, pygidial, rectal, and poison glands do not alter the characteristic aggressive reactions generally performed by resident workers against alien ants. By contrast, the Dufour's gland seems to play a crucial role in the appeasement of residents of the target host colony. In fact, its secretion drastically lowers the degree of overt attacks shown by F. cunicularia workers against the intruders. This chemical strategy probably allows an easier invasion and usurpation of host colonies by newly mated females of P. rufescens.

Plasticity of the Cuticular Hydrocarbon Profile of the Slave-Making Ant Polyergus Rufescens Depending on the Social Environment

Analysis of cuticular hydrocarbon mixtures of the slave-making ant Polyergus rufescens and the Formica rufibarbis or F. cunicularia slaves with which it was living either naturally or after the slave species had been experimentally exchanged showed that cohabitation induced no qualitative homogenization of these species' cuticular signatures. The proportions of hydrocarbons common to both species tended however to be adjusted (in the case of the natural associations) or readjusted (in that of the experimental associations), especially in Polyergus towards those of their partners.On the other hand, differences were observed between the cuticular profiles of Polyergus and their Formica slaves which had been living separately from the original colony for several weeks in an experimental group, and those of the members of the original colony. The changes in the cuticular profiles occurred synchronously in all the individual members of the group.Comparisons between the cuticular versus postpharyngeal slave-maker and slave mixtures showed the existence of qualitative differences between the two mixtures. This indicates that the postpharyngeal glands are not directly involved in the production of the cuticular signatures. Some possible explanations for these results are discussed.

Intercolonial differences in raiding activities in the Japanese slave-making antPolyergus samurai

Intercolonial differences in raiding activity were investigated in the field on 9 colonies of the slavemaking antPolyergus samurai. Duration of raiding season, the mean start time of the first raiding trip in a day, the mean speed of outbound and inbound trips, and the mean distance to target nests varied significantly among colonies. The variance in start time of raiding was correlated with that in soil temperature and diurnal change pattern in soil temperature at the nest sites. The speed of trips and the first day with trips in the season also correlated with soil temperature at the nest site. Simple environmental factors well explained the observed variances in raiding activity among colonies. Therefore, raiding behaviour ofP. samurai seemed to be a stereotyped behaviour that is regulated through simple environmental factors.

Raiding behavior of the Japanese slave-making antPolyergus samurai

Raiding behavior of the Japanese slave-making antPolyergus samurai was investigated in the field. Raiding trips occurred from early June to early September. A raiding column of several hundreds workers would rush into a target nest and rob mainly worker pupae of the host species,Formica (Serviformica) japonica. Most trips occurred on sunny days. Air temperature, soil temperature, relative humidity, and radiation energy at the ground surface were significantly different between days with and without raiding trips. Nuptial flights occurred on hot, sunny days, and mostPolyergus colonies released alates simultaneously. Behaviors of newly mated queens are also provided and are compared with otherPolyergus species.

Specificity in host choice by the slave-making antPolyergus rufescensLatr. (Hymenoptera Formicidae)

The interspecific relationships between the obligatory slave-making ant Polyergus rufescens and three host species belonging to the Formica fusca group (i.e. F. cunicularia, F. gagates, and F. rufibarbis) were analysed and quantified in the laboratory using an aggression test. Fierce and immediate attacks accompanied by the elements typical of overt aggression (i.e. gaster flexing, seizing, dragging, and carrying) characterized the encounters between P. rufescens and two of the host species: F. gagates (90%) and F. rufibarbis (95%). By contrast, only 9 (45%) pairs of slave-maker and F. cunicularia individuals fought and these encounters were always interrupted by mutual antennal inspections accompanied by the items of ritualized aggressive behaviour (i.e. threat and upright posture). The lower degree of aggression between P. rufescens and F. cunicularia compared with that occurring with the other two potential host species, could explain the presence of only F. cunicularia individuals as slaves in the nat...