Colony Recognition Research Articles

The Effect of Diet on Colony Recognition and Cuticular Hydrocarbon Profiles of the Invasive Argentine Ant, Linepithema humile.

Simple SummaryThe membership of social insect colonies is defined by chemical pheromones on the bodies of colony members. In nearly all ant species that have been studied, these pheromones have been shown to be genetically based. In some cases, however, environmentally derived odors have been implicated as colony recognition cues. The widely introduced and invasive Argentine ant is well known for forming massive “supercolonies” in its introduced ranges. Previous studies have implicated both genetic and diet-derived chemicals in the colony recognition systems of introduced populations. Here, we perform feeding experiments, in both realistic field settings and the lab, and show that dietary changes do not cause behavioral changes in the field, as well as under most laboratory conditions. However, one exception was found, in which reduced aggression was recorded in one of the laboratory feeding treatments (with crickets as the dietary item), but, nevertheless, not of sufficient magnitude to explain the unusual colony structure of introduced Argentine ants. We conclude that dietary shifts during introduction to new ranges do not account for the origin of Argentine ant supercolonies.Ants are some of the most abundant and ecologically successful terrestrial organisms, and invasive ants rank among the most damaging invasive species. The Argentine ant is a particularly well-studied invader, in part, because of the extreme social structure, known as unicoloniality, that occurs in introduced populations. Unicoloniality is characterized by the formation of geographically vast supercolonies, within which territorial behavior and intraspecific aggression are absent. Although there is considerable evidence supporting a genetic basis for the odor cues involved in colony recognition, some studies have suggested that diet may also influence colony recognition cues and, thus, colony structure. Here, we test the role for insect-derived recognition cues by performing a diet supplementation experiment in a natural field setting, and a more extreme dietary manipulation experiment in the lab. After one month, in both the field and the lab, we found that aggressive supercolonies remained aggressive toward each other and non-aggressive nests (from the same supercolony) remained non-aggressive, regardless of dietary treatment. In one lab treatment, we did observe a significant decrease in the level of aggression between different supercolonies that were fed the same diet, but aggression was still frequent. We did not see any evidence for cuticular hydrocarbon odor cues being transferred from prey to ants in any of the field treatments. In the more extreme lab treatment, however, several cuticular hydrocarbons were acquired from both roach and cricket insect prey (but not Drosophila). Based on these data, we conclude that dietary changes are unlikely to underlie changes in behavior or colony structure in Argentine ants in real-world settings. However, these results indicate that caution is warranted when interpreting the behaviors of animals that have been reared on diets that are substantially different from natural populations.

Ovary activation correlates with the reproductive potential of honeybee (Apis mellifera) workers if they are in a foreign colony

In eusocial Hymenoptera, the laying of male-determined eggs by workers in an unrelated colony can be a powerful strategy for increasing direct fitness benefits. A recent study showed that honeybee rebels, which are workers that develop under queenless conditions and have high reproductive potential, drift to foreign colonies, with a preference for hopelessly queenless colonies, and act as reproductive parasites. In our experiment, by introducing 5-day-old rebels and normal workers into foreign queenright colonies, we discovered that only rebels developed a reproductive phenotype by activating their ovaries. In a similar experiment with 1-day-old workers, neither rebels nor normal workers displayed such a tendency. We suggest that workers’ reproductive potential could thus be a key parameter activating the ovaries in not only drifting workers but also workers acting as reproductive parasites. Our results also support the hypothesis that the colony recognition cue probably has an environmental, rather than a genetic, origin.

Species-specific signatures of the microbiome from Camponotus and Colobopsis ants across developmental stages.

Symbiotic relationships between hosts and bacteria are common in nature, and these may be responsible for the evolutionary success of various groups of animals. Among ants, these associations have been well studied in some genera of the Camponotini, but several questions remain regarding the generality of the previous findings across all the members of this ant tribe and if bacterial communities change across development in these hosts. This study is the first to characterize the bacterial community associated with a colony of the recently recognized genus Colobopsis and three colonies of Camponotus (two distinct species) and show how different the composition of the bacterial community is when compared across the different genera. Our data reveal that Colobopsis (species: Co. riehlii) and Camponotus (species: Ca. floridanus and Ca. planatus) have distinct microbiota, and we were able to verify that the identity of the species contributes more to the bacterial diversity. We also demonstrated that there were no significant differences between colonies of the same species (Camponotus planatus), and between stages of development from different colonies. We did find that some developmental stages have distinct bacteria, confirming that each stage of development could have a specific microbiota. Our results show species are one of the factors that shape the bacterial community in these Camponotini ants. Additional studies of the intra-colonial microbiome of other hosts and across development may reveal additional clues about the function and importance of bacteria in colony recognition, individual and colony health, and nutritional upgrading.

Outnumbered: a new dominant ant species with genetically diverse supercolonies in Ethiopia

A Lepisiota (Hymenoptera: Formicidae: Formicinae) species in Ethiopia has been observed forming supercolonies spanning up to 38 km. L. canescens occurs at very high densities where there is sufficient moisture or herbaceous cover and dominates the local ant community, traits reminiscent of an invasive species. The supercolonies are genetically diverse, however, indicating they have not gone through the population bottleneck usually characteristic of species invasions. We conclude that the species is native to this region, though expanding its range locally into areas of human disturbance, where it is exploding in numbers. The lack of aggression across a genetically diverse population suggests that mitochondrial genetic variation is decoupled from variation relating to colony recognition cues like cuticular hydrocarbons. All in all, L. canescens could have the makings of an invasive species at an international scale and may represent a novel system to study the evolution and spread of supercolonies in ants.

Two Novel Volatile Compounds as the Key for Intraspecific Colony Recognition in Macrotermes gilvus (Isoptera: Termitidae)

Two Novel Volatile Compounds as the Key for Intraspecific Colony Recognition in Macrotermes gilvus (Isoptera: Termitidae)

Does the volatile hydrocarbon profile differ between the sexes: a case study on five aphidophagous ladybirds.

Insect hydrocarbons (HCs) primarily serve as a waterproofing cuticular layer and function extensively in chemical communication by facilitating species, sex, and colony recognition. In this study, headspace solid-phase microextraction is employed for investigating the sex-specific volatile HC profile of five ladybirds collected from Lucknow, India namely, Coccinella septempunctata (L.), Coccinella transversalis (Fabr.), Menochilus sexmaculatus (Fabr.), Propylea dissecta (Mulsant), and Anegleis cardoni (Weise) for the first time. Major compounds reported in C. septempunctata, C. transversalis, and A. cardoni are methyl-branched saturated HCs, whereas in M. sexmaculatus, and P. dissecta, they are unsaturated HCs. Other than A. cardoni, both the sexes of the other four ladybirds had similar compounds at highest peak but with statistically significant differences. However, in A. cardoni, which is a beetle with a narrow niche, the major compound in both male and female was different. The difference in volatile HC profile of the sexes of the five ladybirds indicates that gender-specific differences primarily exist due to quantitative differences in chemicals with only very few chemicals being unique to a gender. This variation in semiochemicals might have a role in behavioral or ecological aspects of the studied ladybirds.

Dissecting ant recognition systems in the age of genomics.

Hamilton is probably best known for his seminal work demonstrating the role of kin selection in social evolution. His work made it clear that, for individuals to direct their altruistic behaviours towards appropriate recipients (kin), mechanisms must exist for kin recognition. In the social insects, colonies are typically comprised of kin, and colony recognition cues are used as proxies for kinship cues. Recent years have brought rapid advances in our understanding of the genetic and molecular mechanisms that are used for this process. Here, I review some of the most notable advances, particularly the contributions from recent ant genome sequences and molecular biology.

Friends and Foes from an Ant Brain's Point of View – Neuronal Correlates of Colony Odors in a Social Insect

BackgroundSuccessful cooperation depends on reliable identification of friends and foes. Social insects discriminate colony members (nestmates/friends) from foreign workers (non-nestmates/foes) by colony-specific, multi-component colony odors. Traditionally, complex processing in the brain has been regarded as crucial for colony recognition. Odor information is represented as spatial patterns of activity and processed in the primary olfactory neuropile, the antennal lobe (AL) of insects, which is analogous to the vertebrate olfactory bulb. Correlative evidence indicates that the spatial activity patterns reflect odor-quality, i.e., how an odor is perceived. For colony odors, alternatively, a sensory filter in the peripheral nervous system was suggested, causing specific anosmia to nestmate colony odors. Here, we investigate neuronal correlates of colony odors in the brain of a social insect to directly test whether they are anosmic to nestmate colony odors and whether spatial activity patterns in the AL can predict how odor qualities like “friend” and “foe” are attributed to colony odors.Methodology/Principal FindingsUsing ant dummies that mimic natural conditions, we presented colony odors and investigated their neuronal representation in the ant Camponotus floridanus. Nestmate and non-nestmate colony odors elicited neuronal activity: In the periphery, we recorded sensory responses of olfactory receptor neurons (electroantennography), and in the brain, we measured colony odor specific spatial activity patterns in the AL (calcium imaging). Surprisingly, upon repeated stimulation with the same colony odor, spatial activity patterns were variable, and as variable as activity patterns elicited by different colony odors.ConclusionsAnts are not anosmic to nestmate colony odors. However, spatial activity patterns in the AL alone do not provide sufficient information for colony odor discrimination and this finding challenges the current notion of how odor quality is coded. Our result illustrates the enormous challenge for the nervous system to classify multi-component odors and indicates that other neuronal parameters, e.g., precise timing of neuronal activity, are likely necessary for attribution of odor quality to multi-component odors.

The scent of supercolonies: the discovery, synthesis and behavioural verification of ant colony recognition cues

BackgroundAnts form highly social and cooperative colonies that compete, and often fight, against other such colonies, both intra- and interspecifically. Some invasive ants take sociality to an extreme, forming geographically massive 'supercolonies' across thousands of kilometres. The success of social insects generally, as well as invasive ants in particular, stems from the sophisticated mechanisms used to accurately and precisely distinguish colonymates from non-colonymates. Surprisingly, however, the specific chemicals used for this recognition are virtually undescribed.ResultsHere, we report the discovery, chemical synthesis and behavioural testing of the colonymate recognition cues used by the widespread and invasive Argentine ant (Linepithema humile). By synthesizing pure versions of these chemicals in the laboratory and testing them in behavioural assays, we show that these compounds trigger aggression among normally amicable nestmates, but control hydrocarbons do not. Furthermore, behavioural testing across multiple different supercolonies reveals that the reaction to individual compounds varies from colony to colony -- the expected reaction to true colony recognition labels. Our results also show that both quantitative and qualitative changes to cuticular hydrocarbon profiles can trigger aggression among nestmates. These data point the way for the development of new environmentally-friendly control strategies based on the species-specific manipulation of aggressive behaviour.ConclusionOverall, our findings reveal the identity of specific chemicals used for colonymate recognition by the invasive Argentine ants. Although the particular chemicals used by other ants may differ, the patterns reported here are likely to be true for ants generally. As almost all invasive ants display widespread unicoloniality in their introduced ranges, our findings are particularly relevant for our understanding of the biology of these damaging invaders.

Hydrocarbons in the Ant Lasius niger: From the Cuticle to the Nest and Home Range Marking

The cuticular hydrocarbons (CHCs) of the ant Lasius niger are described. We observe a high local colony specificity of the body cuticular profile as predicted for a monogynous and multicolonial species. The CHCs show a low geographical variation among different locations in France. The CHCs on the legs also are colony specific, but their relative quantities are slightly different from those on the main body. For the first time, we demonstrate that the inner walls of the ant nest are coated with the same hydrocarbons as those found on the cuticle but in different proportions. The high amount of inner-nest marking and its lack of colony-specificity may explain why alien ants are not rejected once they succeed in entering the nest. The cuticular hydrocarbons also are deposited in front of the nest entrance and on the foraging arena, with a progressive increase in n-alkanes relative amounts. Chemical marks laid over the substrate are colony specific only when we consider methyl-branched alkanes. Our data confirm that these "footprint hydrocarbons" are probably deposited passively by the contact of ant tarsae with the substrate. These results suggest that the CHCs chemical profiles used by ants in colony recognition are much more complex than a single template: ants have to learn and memorize odors that vary depending on their context of perception.

Hair sheep blood, citrated or defibrinated, fulfills all requirements of blood agar for diagnostic microbiology laboratory tests.

BackgroundBlood agar is used for the identification and antibiotic susceptibility testing of many bacterial pathogens. In the developing world, microbiologists use human blood agar because of the high cost and inhospitable conditions for raising wool sheep or horses to supply blood. Many pathogens either fail to grow entirely or exhibit morphologies and hemolytic patterns on human blood agar that confound colony recognition. Furthermore, human blood can be hazardous to handle due to HIV and hepatitis [1], [2]. This study investigated whether blood from hair sheep, a hardy, low-maintenance variety of sheep adapted for hot climates, was suitable for routine clinical microbiology studies.Methods and FindingsHair sheep blood obtained by jugular venipuncture was anticoagulated by either manual defibrination or collection in human blood bank bags containing citrate-phosphate-dextrose. Trypticase soy 5% blood agar was made from both forms of hair sheep blood and commercial defibrinated wool sheep blood. Growth characteristics, colony morphologies, and hemolytic patterns of selected human pathogens, including several streptococcal species, were evaluated. Specialized identification tests, including CAMP test, reverse CAMP test, and satellite colony formation with Haemophilus influenzae and Abiotrophia defectiva were also performed. Mueller-Hinton blood agar plates prepared from the three blood types were compared in antibiotic susceptibility tests by disk diffusion and E-test.ConclusionsThe results of all studies showed that blood agar prepared from citrated hair sheep blood is suitable for microbiological tests used in routine identification and susceptibility profiling of human pathogens. The validation of citrated hair sheep blood eliminates the labor-intensive and equipment-requiring process of manual defibrination. Use of hair sheep blood, in lieu of human blood currently used by many developing world laboratories and as an alternative to cost-prohibitive commercial sheep blood, offers the opportunity to dramatically improve the safety and accuracy of laboratory diagnosis of pathogenic bacteria in resource-poor countries.

Ant colony recognition systems for part clustering problems

Cellular manufacturing requires an effective part clustering method to start up the manufacturing cell design. This paper presents a new part clustering algorithm that uses the concept of the recognition system of artificial ants. The proposed algorithm mimics the random meetings of real ants to build up the ability of object recognition and then to form many initial part clusters with high similarities. These initial part clusters are further merged into larger and larger clusters in an agglomerative way until the designated number of part families is reached. The characteristics of artificial ants, such as randomization and collective behaviour, allow the algorithm to re-cluster wrongly grouped parts into the proper clusters. As a result this can eliminate the chaining effects resulting from the interference of abnormal parts during the clustering process. This algorithm has been developed into a software system called the ant colony recognition system (ACRS). A number of problems selected from the literature have been solved by ACRS, and the evaluation results indicate that ACRS is able to solve the cell formation problems effectively.

Why are larvae of the social parasite wasp Polistes sulcifer not removed from the host nest?

A challenge for parasites is how to evade the sophisticated detection and rejection abilities of potential hosts. Many studies have shown how insect social parasites overcome host recognition systems and successfully enter host colonies. However, once a social parasite has successfully usurped an alien nest, its brood still face the challenge of avoiding host recognition. How immature stages of parasites fool the hosts has been little studied in social insects, though this has been deeply investigated in birds. We look at how larvae of the paper wasp obligate social parasite Polistes sulcifer fool their hosts. We focus on cuticular hydrocarbons (CHCs), which are keys for adult recognition, and use behavioral recognition assays. Parasite larvae might camouflage themselves either by underproducing CHCs (odorless hypothesis) or by acquiring a chemical profile that matches that of their hosts. GC/MS analyses show that parasite larvae do not have lower levels of CHCs and that their CHCs profile is similar to the host larval profile but shows a reduced colony specificity. Behavioral tests show that the hosts discriminate against alien conspecific larvae from different colonies but are more tolerant towards parasite larvae. Our results demonstrate that parasite larvae have evolved a host larval profile, which overcomes the host colony recognition system probably because of the lower proportion of branched compounds compared to host larvae. In some ways, this is a similar hypothesis to the odorless hypothesis, but it assumes that the parasite larvae are covered by a chemical blend that is not meaningful to the host.

Intraspecific competition in the ant Camponotus cruentatus: should we expect the ‘dear enemy’ effect?

The mechanisms of competition and resource domination were analysed in the Mediterranean ant Camponotus cruentatus. In a plot located in southern Spain mature colonies are overdispersed, which is indicative of territoriality in other species. Unexpectedly, however, workers from neighbouring colonies commonly foraged in areas that overlapped by more than 44%. This suggests that while mature colonies possibly prevent the establishment of new colonies in the vicinity of their nests, they do not limit the intrusion of alien foragers. In contrast, rich food sources located in common foraging areas are aggressively defended, generally leading to their domination by the nest that discovered them first. Resource exploitation further requires rapid recruitment of foragers and soldiers to transport food as well as to exclude competitors. Complementary to the field experiments we conducted laboratory bioassays to assess intercolonial aggression. Group encounters showed that workers of C. cruentatus were equally aggressive towards alien ants irrespective of whether they were sympatric neighbours (and had possibly had contact), sympatric nonneighbours (which had probably had no contact), or allopatric. This was corroborated by analysis of the hydrocarbons that generally serve as colony recognition cues. This mixture, which contains an unusual proportion of trimethylalkanes, showed important colony specificity. The high intraspecific aggression shown indiscriminately by workers indicates that the ‘dear enemy’ effect does not exist in this species and suggests that scarce resources are worth defending against intraspecific competitors. We discuss several proximate and ultimate constraints that may prevent the general occurrence of this process in ants.

Nestmate recognition and intraspecific chemical and genetic variation in Temnothorax ants

The evolutionary stability of social cooperation requires altruistic acts to be directed mainly towards related individuals. To maintain significant intragroup relatedness, social insects have evolved sophisticated recognition systems. We compared nestmate recognition abilities of three native European species of the ant genus Temnothorax in relation to variation in cuticular chemicals and intraspecific genetic diversity. Colony take-over and fusions are common in two sibling species T. nylanderi and T. crassispinus, but rare in T. unifasciatus, suggesting relatively inefficient nestmate recognition in the former species. Temnothorax nylanderi and T. crassispinus discriminated less effectively between nestmates and non-nestmates than T. unifasciatus. While chemical profiles of different colonies strongly overlapped in T. crassispinus and T. nylanderi, 97% of all T. unifasciatus samples were, in a discriminant analysis, correctly assigned to their respective colony. Chemical distances between colonies correlated with aggression levels in T. unifasciatus, but not in the other two species. Finally, preliminary genetic results indicated that haplotype divergence within T. nylanderi and T. crassispinus is remarkably low, whereas T. unifasciatus and other congeners are genetically more variable. In theory, loss of genetic variation, including depletion at loci involved in the production of cuticular hydrocarbons, could decrease colony recognition efficiency. We suggest that this might be the case in T. nylanderi and T. crassispinus, leading to colony fusions in the field. Despite their different life histories, these native species show similarities to invasive ants where genetic bottlenecks arising from founder effects are thought to result in a loss of colony boundaries.

The Mandibular Gland Secretions of the Leaf-Cutting Ants Atta sexdens sexdens and Atta opaciceps Exhibit Intercaste and Intercolony Variations

The mandibular gland secretions of worker castes from wild colonies of the leaf-cutting ants Atta sexdens sexdens and Atta opaciceps were analyzed quantitatively by gas chromatography-mass spectrometry. The secretions comprised a complex mixture of volatile, mainly oxygenated compounds, and their profiles exhibited considerable qualitative and quantitative variations among species and castes. The known alarm pheromone 4-methyl-3-heptanone was common to both species. The elevated relative proportions of this ketone found in the secretions of gardeners and generalists suggest that such castes are primarily responsible for the production and release of the alarm pheromone. Quantitative variations (but no qualitative differences) in the profiles of secretions of soldiers from different colonies of A. sexdens sexdens were detected, supporting the view that intraspecific colony recognition is mediated through mandibular gland secretions. Subsequent laboratory assays showed that, among the compounds identified by GC-MS, 4-methyl-3-heptanone elicited a strong alarm response in workers of A. sexdens sexdens and A. opaciceps.

Optimal detection of Staphylococcus aureus from clinical specimens using a new chromogenic medium

The new chromogenic medium CHROMagar Staph aureus (CASA) was evaluated for its ability to detect and presumptively identify Staphylococcus aureus. Nine hundred forty-two clinical specimens (742 wound, 200 sputum and bronchoalveolar lavage) were cultured on CASA, tryptic soy blood agar (TSBA), and mannitol salt agar (MSA). Of the 153 S. aureus isolates from wounds on any media, 151 grew on CASA and TSBA and 146 on MSA. Sensitivity after 24 hours was 93.5%, 94%, and 77%, respectively, and increased after 48 hours to 99% for CASA and TSBA and to 95% for MSA. Of the 41 isolates recovered from sputum and lavage, all grew on CASA, 27 on TSBA, and 36 on MSA. Sensitivity after 24 hours was 93%, 66%, and 81%, respectively, and 100%, 66%, and 88% after 48 hours. All specimens revealed 99% sensitivity for CASA, 92% for TSBA, and 94% for MSA. Specificity for CASA was 100%. Antimicrobial susceptibility tests showed full agreement between isolates from CASA and from reference media. In conclusion, CASA has a high sensitivity and can identify isolates undetected on conventional media ( p value for CASA vs. TSBA was 0.001 and vs. MSA, 0.006). This difference is particularly notable when mixed flora are present. The simplicity of the colony recognition increased the medium specificity, allowing a reliable and rapid method for the detection of S. aureus on the primary plate.

Cuticular hydrocarbons and aggression in the termite Macrotermes subhyalinus.

Cuticular hydrocarbons are among the prime candidates for nestmate recognition in social insects. We analyzed the variation of cuticular hydrocarbons in the termite species M. subhyalinus in West Africa (Comoë National Park) on a small spatial scale (< 1 km). We found considerable variation in the composition of cuticular hydrocarbons among colonies, with four distinct chemical phenotypes. Different phenotypes occurred within each of the four habitats. The difference between these phenotypes is primarily due to unsaturated compounds. A clear correlation between the difference of the hydrocarbon composition and the aggression between colonies was found. This correlation also holds in a multivariate analysis of genetic similarity (measured by AFLPs), morphometric distances (measured by Mahalanobis-distances). as well as geographic distances between colonies. In a more detailed analysis of the correlation between the composition of cuticular hydrocarbons and aggression, we found that no single compound is sufficient to explain variation in aggression between pairings of colonies. Thus, termites seem to use a bouquet of compounds. Multiple regression analysis suggested that many of these compounds are unsaturated hydrocarbons and, thus, may play a key role in colony recognition.

Testing the blank slate hypothesis: why honey bee colonies accept young bees

Special features facilitate the admission of new members, such as neonates, to otherwise closed animal societies. In eusocial insects, such as honeybees and paper wasps, young adults acquire a colony recognition phenotype from other colony members or nesting materials. Older adults must exempt them from expulsion during the acquisition period. Newly emerged adult honeybees gain tolerance in their colony before their acquisition of the colony recognition phenotype by presenting a blank slate, absent recognition cues. This makes them generically acceptable in honey bee colonies. This strategy is analogous to the easily recognizable phenotypes associated with juvenility in birds and mammals.

Colony insularity through queen control on worker social motivation in ants.

We investigated the relative contribution of the queen and workers to colony nestmate recognition cues and on colony insularity in the Carpenter ant Camponotus fellah. Workers were either individually isolated, preventing contact with both queen and workers (colonial deprived, CD), kept in queenless groups, allowing only worker-worker interactions (queen deprived, QD) or in queenright (QR) groups. Two weeks post-separation QD and QR workers were amicable towards each other but both rejected their CD nestmates, which suggests that the queen does not measurably influence the colony recognition cues. By contrast, aggression between QD and QR workers from the same original colony was apparent only after six months of separation. This clearly demonstrates the power of the Gestalt and indicates that the queen is not a dominant contributor to the nestmate recognition cues in this species. Aggression between nestmates was correlated with a greater hydrocarbon (HC) profile divergence for CD than for QD and QR workers, supporting the importance of worker-worker interactions in maintaining the colony Gestalt odour. While the queen does not significantly influence nestmate recognition cues, she does influence colony insularity since within 3 days QD (queenless for six months) workers from different colony origins merged to form a single queenless colony. By contrast, the corresponding QR colonies maintained their territoriality and did not merge. The originally divergent cuticular and postpharyngeal gland HC profiles became congruent following the merger. Therefore, while workers supply and blend the recognition signal, the queen affects worker-worker interaction by reducing social motivation and tolerance of alien conspecifics.