Nestmate recognition in ants is possible without tactile interaction

Ants of the genus Camponotus are able to discriminate recognition cues of colony members (nestmates) from recognition cues of workers of a different colony (non-nestmates) from a distance of 1 cm. Free moving, individual Camponotus floridanus workers encountered differently treated dummies on a T-bar and their behavior was recorded. Aggressive behavior was scored as mandibular threat towards dummies. Dummies were treated with hexane extracts of postpharyngeal glands (PPGs) from nestmates or non-nestmates which contain long-chain hydrocarbons in ratios comparable to what is found on the cuticle. The cuticular hydrocarbon profile bears cues which are essential for nestmate recognition. Although workers were prevented from antennating the dummies, they showed significantly less aggressive behavior towards dummies treated with nestmate PPG extracts than towards dummies treated with non-nestmate PPG extracts. In an additional experiment, we show that cis-9-tricosene, an alkene naturally not found in C. floridanus' cuticular profile, is behaviorally active and can interfere with nestmate recognition when presented together with a nestmate PPG extract. Our study demonstrates for the first time that the complex multi-component recognition cues can be perceived and discriminated by ants at close range. We conclude that contact chemosensilla are not crucial for nestmate recognition since tactile interaction is not necessary.     

An absence of aggression between non-nestmates in the bull ant <Emphasis Type="Italic">Myrmecia nigriceps</Emphasis>

The ability of social insects to discriminate against non-nestmates is vital for maintaining colony integrity, and in most social insect species, individuals act aggressively towards non-nestmates that intrude into their nest. Our experimental field data revealed that intra-colony aggression in the primitive bulldog ant Myrmecia nigriceps is negligible; our series of bioassays revealed no significant difference in the occurrence of aggression in trials involving workers from the same, a close (less than 300 m) or a far (more than 1.5 km) nest. Further, non-nestmate intruders were able to enter the nest in 60% of our trials; a similar level was observed in trials involving nestmates. These results suggest that workers of M. nigriceps are either unable to recognize alien conspecifics or that the costs of ignoring workers from foreign colonies are sufficiently low to favor low levels of inter-colony aggression in this species.     

Nestmate recognition in social wasps: manipulation of hydrocarbon profiles induces aggression in the European hornet

The influence of individual cuticular hydrocarbons on nestmate recognition in the European hornet, Vespa crabro L., was investigated. We observed the behavioural response of workers towards differently treated dead conspecifics in a bioassay. Dummies were extracted with dichloromethane and extracts were spiked with microgram amounts of synthetic hydrocarbons naturally occurring on the cuticle of V. crabro. These modified extracts were reapplied to extracted workers that were subsequently tested in the bioassay. Non-spiked nestmate dummies (negative control) and untreated non-nestmate dummies (positive control) were tested in control experiments. The addition of only heneicosane or a mixture of heneicosane, tricosane, and (Z)-9-tricosene to the extracts led to a significant increase of agonistic behaviour in workers leaving the nest for foraging flights. Returning workers reacted much less aggressively than those leaving. This is one of the first behavioural proofs that manipulation of cuticular hydrocarbon profiles can be perceived by a social insect species. The results support the hypothesis that colony-specific cuticular hydrocarbon profiles are involved in the phenomenon of nestmate recognition among social insects.     

Irregular brood patterns and worker reproduction in social wasps

The potential for reproductive conflict among colony members exists in all social insect societies. For example, queens and workers may be in conflict over the production of males within colonies. Kin selection theory predicts that in a colony headed by a multiply-mated queen, worker reproduction is prevented by worker policing in the form of differential oophagy. However, few studies have demonstrated that workers actually lay eggs within queenright colonies. The purpose of this study was to determine if workers laid male eggs within unmanipulated queen-right colonies of the polyandrous social wasps Vespula maculifrons and V. squamosa. We focused our analysis on an unusual brood pattern within colonies, multiple egg cells. We were primarily interested in determining if individuals reared in these irregular circumstances were queen or worker offspring. To address this question, we genotyped 318 eggs from eight V. maculifrons and two V. squamosa colonies. No worker‑reproduction was detected in any of the queenright colonies; all of the eggs found in multiple egg cells were consistent with being queen‑produced. However, the frequency of multiple egg cells differed among colonies, suggesting that queens vary in the frequency of errors they make when laying eggs within cells. Finally, we suggest that workers may not be laying eggs within queenright colonies and that worker reproduction may be controlled through mechanisms other than differential oophagy in polyandrous Vespula wasps.     

Nest- and colony-mate recognition in polydomous colonies of meat ants (Iridomyrmex purpureus)

Workers of polydomous colonies of social insects must recognize not only colony-mates residing in the same nest but also those living in other nests. We investigated the impact of a decentralized colony structure on colony- and nestmate recognition in the polydomous Australian meat ant (Iridomyrmex purpureus). Field experiments showed that ants of colonies with many nests were less aggressive toward alien conspecifics than those of colonies with few nests. In addition, while meat ants were almost never aggressive toward nestmates, they were frequently aggressive when confronted with an individual from a different nest within the same colony. Our chemical analysis of the cuticular hydrocarbons of workers using a novel comprehensive two-dimensional gas chromatography technique that increases the number of quantifiable compounds revealed both colony- and nest-specific patterns. Combined, these data indicate an incomplete transfer of colony odor between the nests of polydomous meat ant colonies.     

“You are what you eat”: Diet modifies cuticular hydrocarbons and nestmate recognition in the Argentine ant, Linepithema humile

 Nestmate recognition plays a key role in the behavior and evolution of social insects. We demonstrated that hydrocarbons are the chemical cues used in Argentine ant, Linepithema humile, nestmate recognition, and that these hydrocarbons can be acquired from insect prey. Consequently, Argentine ant cuticular hydrocarbon patterns reveal the same hydrocarbons present in their diet. Diet alters both the recognition cues present on the cuticular surface and the response of nestmates to this new colony odor, resulting in aggression between former nestmates reared on different insect prey. Received: 16 June 2000 / Accepted: 14 August 2000     

Sensing the intruder: a quantitative threshold for recognition cues perception in honeybees

The ability to discriminate among nestmates and non-nestmate is essential to defend social insect colonies from intruders. Over the years, nestmate recognition has been extensively studied in the honeybee Apis mellifera; nevertheless, the quantitative perceptual aspects at the basis of the recognition system represent an unexplored subject in this species. To test the existence of a cuticular hydrocarbons’ quantitative perception threshold for nestmate recognition cues, we conducted behavioural assays by presenting different amounts of a foreign forager’s chemical profile to honeybees at the entrance of their colonies. We found an increase in the explorative and aggressive responses as the amount of cues increased based on a threshold mechanism, highlighting the importance of the quantitative perceptual features for the recognition processes in A. mellifera.     

Scent of a queen—cuticular hydrocarbons specific for female reproductives in lower termites

In social insects, it is assumed that signals of the queen inform nestmates about her reproductive status. Thus, workers forego their own reproduction if the queen signals high fertility. In hemimetabolous termites, little is known about reproductive inhibition, but evidence exists for a royal-pair control. Workers of lower termites exhibit a high developmental flexibility and are potentially able to become reproductives, but the presence of a fertile reproductive restrains them from reaching sexual maturity. The nature of this control, however, remains unknown. Here, we report on qualitative differences in cuticular hydrocarbon profiles between queens and workers of the basal drywood termite Cryptotermes secundus. Queens were characterized by a shift to long-chained and branched hydrocarbons. Most remarkably, similar chemical patterns are regarded as fertility cues of reproductives in social Hymenoptera. This might suggest that both groups of social insects convergently evolved similar chemical signatures. The present study provides deeper insights into how termites might have socially exploited these signatures from sexual communication in their cockroach-like ancestor. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Queen regulates biogenic amine level and nestmate recognition in workers of the fire ant, <Emphasis Type="Italic">Solenopsis invicta</Emphasis>

Nestmate recognition is a critical element in social insect organization, providing a means to maintain territoriality and close the colony to parasites and predators. Ants detect the colony chemical label via their antennae and respond to the label mismatch of an intruder with aggressive behavior. In the fire ant, Solenopsis invicta, worker ability to recognize conspecific nonnestmates decreases if the colony queen is removed, such that they do not recognize conspecific nonnestmates as different. Here, we tested the hypothesis that the presence of the colony queen influences the concentration of octopamine, a neuromodulator, in worker ants, which in turn has an effect on nestmate recognition acuity in workers. We demonstrate that queenless workers exhibit reduced brain octopamine levels and reduced discriminatory acuteness; however, feeding queenless workers octopamine restored both. Dopamine levels are influenced by honeybee queen pheromones; however, levels of this biogenic amine were unchanged in our experiments. This is the first demonstration of a link between the presence of the colony queen, a worker biogenic amine, and conspecific nestmate recognition, a powerful expression of colony cohesion and territoriality.     

Factors influencing survival duration and choice of virgin queens in the stingless bee Melipona quadrifasciata

In Melipona quadrifasciata, about 10 % of the females develop into queens, almost all of which are killed. Occasionally, a new queen replaces or supersedes the mother queen or heads a new colony. We investigated virgin queen fate in queenright and queenless colonies to determine the effects of queen behaviour, body mass, nestmate or non-nestmate status, queenright or queenless colony status, and, when queenless, the effect of the time a colony had been queenless, on survival duration and acceptance. None of 220 virgin queens observed in four observation hives ever attacked another virgin queen nor did any of 88 virgin queens introduced into queenright colonies ever attack the resident queen. A new queen was only accepted in a queenless colony. Factors increasing survival duration and acceptance of virgin queens were to emerge from its cell at 2 h of queenlessness, to hide, and to avoid fights with workers. In this way, a virgin queen was more likely to be available when a colony chooses a new queen, 24-48 h after resident queen removal. Running, walking or resting, antennating or trophallaxis, played little or no role, as did the factors body mass or nestmate. “Queen choice” took about 2 h during which time other virgin queens were still being killed by workers. During this agitated process, the bees congregated around the new queen. She inflated her abdomen and some of the workers deposited a substance on internal nest surfaces including the glass lid of the observation hive.     

An epicuticular multilayer reflector generates the iridescent coloration in chrysidid wasps (Hymenoptera, Chrysididae)

Chrysidid wasps in the subfamily Chrysidinae are brood parasitoids or cleptoparasites of other insects and famous for their cuticular iridescence. In this study, we examine the dorsal abdominal cuticle of the chrysidid wasp Hedychrum rutilans to identify the underlying color mechanism. Using scanning electron microscopy, reflectance spectral analysis, and theoretical calculations, we demonstrate the presence of an epicuticular multilayer reflector consisting of six lamellae with a thickness of 185 nm each. The lamellae exhibit a rough surface probably functioning as spacers between the individual layers. The reflector has a measured reflectance maximum at λ = 630 nm, i.e., in the red part of the visible spectrum of light at normal incidence and the reflectance maximum shifts to green as the angle of incidence increases. Complementary theoretical modeling corroborates the view that the epicuticular multilayer generates the iridescent color of the chrysidid cuticle. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Clonal structure affects the assembling behavior in the Japanese queenless ant <Emphasis Type="Italic">Pristomyrmex punctatus</Emphasis>

The queenless ant Pristomyrmex punctatus (Hymenoptera: Myrmicinae) has a unique society that differs from those of other typical ants. This species does not have a queen, and the workers lay eggs and produce their clones parthenogenetically. However, a colony of these ants does not always comprise members derived from a single clonal line. In this study, we examined whether P. punctatus changes its “assembling behavior” based on colony genetic structure. We prepared two subcolonies—a larger one comprising 200 individuals and a smaller one comprising 100 individuals; these subcolonies were established from a single stock colony. We investigated whether these subcolonies assemble into a single nest. The genetically monomorphic subcolonies (single clonal line) always fused into a single nest; however, the genetically polymorphic subcolonies (multiple clonal lines) did not tend to form a single colony. The present study is the first to demonstrate that the colony genetic structure significantly affects social viscosity in social insects.     

Colony disassociation following diet partitioning in a unicolonial ant

Discriminating nestmates from alien conspecifics via chemical cues is recognized as a critical element in maintaining the integrity of insect societies. We determined, in laboratory experiments, that nestmate recognition in an introduced population of the Argentine ant, Linepithema humile, is modified by hydrocarbons acquired from insect prey, and that workers from spatially isolated colony fragments, each provided with prey that possessed distinct cuticular hydrocarbons, displayed aggressive behavior towards their former nestmates. Isolation for 28 days or more between colony fragments fed different prey was sufficient to prevent re-establishment of inter-nest communication for at least an additional 28 days through the introduction of a bridge between the nests. Ants possessed intrinsic cuticular hydrocarbons plus only those hydrocarbons from the prey they received during the isolation period. Colony fragments which were isolated for less than 28 days reunited with workers possessing both prey hydrocarbons. Therefore, L. humile nestmate recognition may be dynamic, being in part dependent on the spatio-temporal distribution of prey, along with physical factors permitting or restricting access of subcolony units to those prey.     

Costs and benefits of subordinate queens in colonies of the ant,<Emphasis Type="Italic"> Leptothorax gredleri</Emphasis>

Queens of some species of social insects form dominance hierarchies in which only the top-ranking individual reproduces. Such unequal partitioning of reproduction can be stable when subordinate queens increase their inclusive fitness indirectly by helping to rear a related dominant queen's offspring. We investigated whether subordinate queens of the ant, Leptothorax gredleri, affect the reproductive success of the dominant queen. In laboratory colonies with two queens, first eggs were laid approximately 20 days later than in colonies with a single queen, presumably due to aggressive competition among nestmate queens. Nevertheless, two-queen colonies produced on average more brood than one-queen colonies, although egg laying was completely or almost completely monopolized by one single queen. The presence of a subordinate queen therefore appears to increase the productivity of the dominant.     

In-nest environment modulates nestmate recognition in the ant<Emphasis Type="Italic"> Camponotus fellah</Emphasis>

Multiple behavioral and chemical studies indicate that ant nestmate recognition cues are low-volatile substances, in particular hydrocarbons (HCs) located on the cuticular surface. We tested the hypothesis that in the ant Camponotus fellah, nest environment, in particular nest volatile odors, can modulate nestmate-recognition-mediated aggression. Workers were individually confined within their own nest in small cages having either a single mesh (SM = limited physical contact permitted) or a double mesh (DM = exposed to nest volatiles only) screen. Individual workers completely isolated outside their nest (CI) served as control. When reintroduced into a group of 50 nestmates, the CI workers were attacked as alien ants after only 2 weeks of separation, whereas the SM workers were treated as nestmates even after 2 months of separation. Aggression towards DM ants depended on the period of isolation. Only DM workers isolated for over 2 months were aggressed by their nestmates, which did not significantly differ from the CI nestmates. Cuticular HC analyses revealed that the profile of the non-isolated ants (NI) was clearly distinct from that of CI, SM and DM ants. Profile differences matched the aggressive response in the case of CI ants but were uncorrelated in the case of SM or DM ants. This suggests that keeping the ants within the nest environment affected nestmate recognition in additional ways than merely altering their HC profile. Nest environment thus appears to affect label–template mismatch by modulating aggressive behavior, as well as the direction at which cuticular HCs diverged during the separation period.     

A unique strategy of host colony exploitation in a parasitic ant: workers of<Emphasis Type="Italic"> Polyrhachis lama</Emphasis> rear their brood in neighbouring host nests

The formicine ant Polyrhachis lama is a social parasite, exploiting its ponerine host ant species Diacamma sp. In most social parasitic associations, the parasitic species are closely related to their host species group, evolving directly from independent ancestors of the host species. However, in the Polyrhachis lama–Diacamma sp. association, the associated species belong to different ant subfamilies. Based on preliminary field surveys, we had presumed that P. lama might have given up its reproductive division of labour, i.e. workers would be able to produce males as well as workers and females parthenogenetically. In this study, this hypothesis was disproved: Polyrhachis lama workers cannot be fertilized and are only able to produce males. In the host–parasite association originating from nests possessing a P. lama queen, workers penetrate surrounding Diacamma sp. nests, carrying brood for rearing within these satellite nests. In this peculiar way, a single P. lama colony is able to exploit several Diacamma sp. colonies simultaneously.     

Disease and colony foundation in the dampwood termite Zootermopsis angusticollis: The survival advantage of nestmate pairs

To determine the impact of inbreeding and outbreeding on disease resistance and survival during colony foundation, nestmate (NM) and non-nestmate (NON) primary reproductives of the dampwood termite Zootermopsis angusticollis were exposed to a single or double dose of conidia of the entomopathogenic fungus Metarhizium anisopliae. Male and female primary reproductive pairs originating from the same parent colony had higher survivorship than NON pairs in control and conidia-exposure treatments. The survival advantage of NM primary reproductives increased with the intensity of pathogen challenge and was significantly greater in the single- and double-dose treatments than in the controls. Although NM pairs had significantly lower mortality than NON pairs, the survivorship of colonies stabilized as they matured and inbred and outbred colonies did not differ in offspring production. These results demonstrate that colony foundation by NON male and female reproductives may have a disease-related survival cost during this critical phase of their life cycle. There may also be a cost associated with lower offspring heterozygosity, but in the first generation this does not appear to significantly impact colony growth.     

Discrimination of flicker frequency rates in the reptile tuatara (Sphenodon )

By investigating the mechanisms that underlie the perception of environmental cues, we may begin to understand how the sensory system governs behavioral responses. This is the first empirical study to examine learning and visual sensitivity in a reptile species, the tuatara (Sphenodon punctatus). We established a non-intrusive psychophysical method by employing an instrumental paradigm in order to examine discrimination learning and the ability to distinguish different flicker frequencies in the tuatara. Seventeen tuatara were trained under an operant conditioning task to respond to various discriminative stimuli flickering between 2.65 and 65.09 Hz. Tuatara were able to learn the operant task and discriminate between a constant light and flicker frequency rates between 2.65 and 45.61 Hz, but not at 65.09 Hz. We demonstrated a reliable psychophysical method where these reptiles could learn a basic operant task and discriminate visual stimuli in the form of flicker frequency rates. The tuatara’s ability to perceive flickering light is comparable to that of avian, mammalian, and other reptilian species. This method is thus suitable for more comprehensive examinations of vision and additional sensory abilities in other reptiles. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Oviposition by small hive beetles elicits hygienic responses from Cape honeybees

Two novel behaviours, both adaptations of small hive beetles (Aethina tumida Murray) and Cape honeybees (Apis mellifera capensis Esch.), are described. Beetles puncture the sides of empty cells and oviposit under the pupae in adjoining cells. However, bees detect this ruse and remove infested brood (hygienic behaviour), even under such well-disguised conditions. Indeed, bees removed 91% of treatment brood (brood cells with punctured walls caused by beetles) but only 2% of control brood (brood not exposed to beetles). Only 91% of treatment brood actually contained beetle eggs; the data therefore suggest that bees remove only that brood containing beetle eggs and leave uninfected brood alone, even if beetles have accessed (but not oviposited on) the brood. Although this unique oviposition strategy by beetles appears both elusive and adaptive, Cape honeybees are able to detect and remove virtually all of the infested brood.     

Reclaiming the crown: queen to worker conflict over reproduction in <Emphasis Type="Italic">Aphaenogaster cockerelli</Emphasis>

In many social taxa, reproductively dominant individuals sometimes use aggression to secure and maintain reproductive status. In the social insects, queen aggression towards subordinate individuals or workers has been documented and is predicted to occur only in species with a small colony size and a low level of queen–worker dimorphism. We report queen aggression towards reproductive workers in the ant species Aphaenogaster cockerelli, a species with a relatively large colony size and a high level of reproductive dimorphism. Through analysis of cuticular hydrocarbon profiles, we show that queens are aggressive only to reproductively active workers. Non-reproductive workers treated with a hydrocarbon typical for reproductives are attacked by workers but not by queens, which suggests different ways of recognition. We provide possible explanations of why queen aggression is observed in this species.