Nestmate recognition signals of the leaf-cutting ant Atta laevigata

Behavioral tests with field colonies of Atta laevigata were performed in order to identify the source of the odors used in nestmate recognition. We tested the postpharyngeal (PPG) and mandibular glands (MG) as putative organs producing chemical signals for nestmate recognition. Chemical analyses of PPG were also undertaken. With a series of bioassays, we confirmed that nestmate recognition is based on cephalic odors and that these odors come mainly from the mandibular gland secretion. We show chemical evidence that odors from MG are dispersed all over the cuticle. Although odors from PPG elicited colony-specific behavioral responses, the types of behaviors they elicited differed from those of nestmate recognition of whole ants or MG extracts. PPG secretion was characterized by long-chain alkanes and methyl branched alkanes of low volatility, whereas MG contained volatile ketones and alcohols.     

Chemical Profiles of Two Pheromone Glands Are Differentially Regulated by Distinct Mating Factors in Honey Bee Queens (Apis mellifera L.)

Pheromones mediate social interactions among individuals in a wide variety of species, from yeast to mammals. In social insects such as honey bees, pheromone communication systems can be extraordinarily complex and serve to coordinate behaviors among many individuals. One of the primary mediators of social behavior and organization in honey bee colonies is queen pheromone, which is produced by multiple glands. The types and quantities of chemicals produced differ significantly between virgin and mated queens, and recent studies have suggested that, in newly mated queens, insemination volume or quantity can affect pheromone production. Here, we examine the long-term impact of different factors involved during queen insemination on the chemical composition of the mandibular and Dufour''s glands, two of the major sources of queen pheromone. Our results demonstrate that carbon dioxide (an anesthetic used in instrumental insemination), physical manipulation of genital tract (presumably mimicking the act of copulation), insemination substance (saline vs. semen), and insemination volume (1 vs. 8 µl) all have long-term effects on mandibular gland chemical profiles. In contrast, Dufour''s gland chemical profiles were changed only upon insemination and were not influenced by exposure to carbon dioxide, manipulation, insemination substance or volume. These results suggest that the chemical contents of these two glands are regulated by different neuro-physiological mechanisms. Furthermore, workers responded differently to the different mandibular gland extracts in a choice assay. Although these studies must be validated in naturally mated queens of varying mating quality, our results suggest that while the chemical composition of Dufour''s gland is associated with mating status, that of the mandibular glands is associated with both mating status and insemination success. Thus, the queen appears to be signaling both status and reproductive quality to the workers, which may impact worker behavior and physiology as well as social organization and productivity of the colony.     

Multiple glandular origins of queen pheromones in the fire ant Solenopsis invicta

The poison sac of the fire ant Solenopsis invicta is the only identified glandular source of pheromones produced by a functional ant queen. This structure, which contains the poison gland, has previously been shown to be the source of a releaser pheromone that mediates queen recognition and tending by workers. The poison sac has also been demonstrated to be the source of a primer pheromone that inhibits winged, virgin queens from shedding their wings (dealating) and developing their ovaries. To determine if the poison sac was the only source of these pheromones, we excised the poison sac from queens and observed whether operated queens retained their pheromonal effects. In a first experiment, the poison sac was removed from functional (egg-laying) queens which were then paired with unoperated nestmate queens in small colonies. Counts of the workers surrounding each queen two weeks after the operation showed that queens without poison sac were as effective as their unoperated nestmates in attracting worker retinues. In a second experiment, we removed the poison sacs of virgin queens which had not yet begun laying eggs and thus had not begun producing queen pheromone. After allowing them to develop their ovaries, these individuals produced amounts of queen recognition pheromone comparable to those secreted by unoperated or sham operated virgin queens as determined by bioassay. Testing the head, thorax and abdomens of functional queens separately revealed that the head was the most attractive region in relation to its relative surface area. Bioassays of extracts of two cephalic glands-the mandibular gland and postpharyngeal gland-showed that the postpharyngeal gland is a second source of the queen recognition pheromone. Finally, we found that virgin queens whose poison sacs were removed before they began producing queen pheromone initiated production of a primer pheromone that inhibits winged virgin queens from dealating, indicating that this pheromonal effect also has an additional but as yet undetermined source. These results parallel those on the honey bee in which several of the pheromonal effects of functional queens appear to have multiple glandular sources.     

Reproductive development and ontogeny of queen pheromone production in the fire ant Solenopsis invicta

Queens of the fire ant, Solenopsis invicta Buren, produce several releaser and primer pheromones. Using bioassays, the ontogeny of three of these pheromones related to reproductive development was investigated. Virgin queens, in which the process of wing-shedding (dealation) serves as an indicator of the initiation of reproductive development, were studied. First, the production of two queen pheromones produced in the poison gland was examined. The pheromone responsible for initiation and maintenance of retinue formation, a releaser effect, was found to be produced in detectable quantities 2 days after dealation, at which time queens showed significant ovary development and many (30%) had started laying eggs. A primer pheromone that inhibits alate virgin queens from dealating was detected in queens 3 days following wing-shedding, when 80% of the queens were ovipositing. Second, I examined the onset of a pheromone of unknown glandular origin produced by reproductively active virgin queens which leads to their destruction in queenright colonies by stimulating workers to attack and kill them. This pheromone is secreted in quantities detectable by bioassay 2 days after dealation. Thus, in S. invicta, the ontogeny of three distinct queen pheromones is tightly linked with ovary development and initiation of egg laying. These results demonstrate reproductive and communicative functions are closely associated during the transition from potential to functional queen.     

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.     

Bioassay design and length of time in the laboratory affect intercolonial interactions of the Formosan subterranean termite (Isoptera, Rhinotermitidae)

This study examined the effect of diet, experimental design, and length of time in the laboratory on intercolonial agonism among Formosan subterranean termite, Coptotermes formosanus Shiraki, colonies. In pairings of 12 C. formosanus Shiraki colonies collected in an urban forest, there was no significant reduction in survival of termites in 30 out of 59 colony pairs compared to colony controls, but there was <50% survival in 18 colony pairs and <10% survival in six colony pairs. There was no correlation between the level of aggressive behavior and the laboratory diet of the termites. Effect of bioassay design and length of time in the laboratory was evaluated in three colony pairs where tests were first conducted on the day of field collection, then colony pairs were retested every 7 days. Aggressive behavior decreased over time in both bioassays, but it tended to decrease more rapidly in the Petri dish tests. The rapid loss of agonism in groups of termites kept in the laboratory demonstrates that changes in environmental factors affect intercolonial agonism. This article presents the results of research only. Mention of a commercial or proprietary product does not constitute endorsement or recommendation by the USDA.     

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.     

Polyethism and nestmate recognition in the alarm reaction of Atta leaf-cutting ants

Abstract.  Ethological studies are conducted under laboratory conditions using workers of Atta sexdens sexdens and Atta opaciceps collected from field colonies to investigate the mechanisms involved in the alarm response and intra-specific recognition in leaf-cutting ants. Hexane extracts from the heads of gardeners and generalist workers elicit higher levels of alarm response in foragers from the same colony than do mandibular gland extracts from foragers and soldiers, indicating that gardeners, generalists and foragers are primarily responsible for the production of alarm pheromone. Foragers subjected to extracts from non-nestmates exhibit significantly greater alarm responses than are induced by similar extracts derived from nestmate workers, suggesting that the alarm pheromone may have a role in nestmate recognition.     

Significance of digging behavior to mortality of red imported fire ant workers, Solenopsis invicta, in fipronil-treated sand

The effect of fipronil-treated sand on digging behavior and mortality of red imported fire ant, Solenopsis invicta Buren, workers was examined in the laboratory. No-choice digging bioassays where fipronil-treated sand was the only available digging substrate were conducted on two colonies at fipronil concentrations of 0.00, 0.05, 0.10, 0.50, 1.00, 1.50, and 2.00 ppm. Workers dug into the fipronil-treated sand in all cases, even at 2.0 ppm level, which caused 100% mortality in acute toxicity tests for both colonies. At 1.5 and 2.0 ppm, workers from the less sensitive colony had significantly higher mortality than those from the more sensitive colony, which might be explained by the significantly higher digging activity of the less sensitive colony. In two-choice digging bioassays where untreated sand was also available, workers dug into the fipronil-treated sand in 29 of 30 cases, even at 10.0 ppm level. At 1.0 and 10.0 ppm, mortality was positively correlated to digging effort in treated sand; however, such correlation was significant only at 1.0 ppm level. This indicates that digging did affect mortality; however, such effect is concentration dependent.     

Transfer of intracolonial genetic variability through gametes in Acropora hyacinthus corals

In recent years, the new phenomenon of intracolonial genetic variability within a single coral colony has been described. This connotes that coral colonies do not necessarily consist of only a single genotype, but may contain several distinct genotypes. Harboring more than one genotype could improve survival under stressful environmental conditions, e.g., climate change. However, so far it remained unclear whether the intracolonial genetic variability of the adult coral is also present in the gametes. We investigated the occurrence of intracolonial genetic variability in 14 mature colonies of the coral Acropora hyacinthus using eight microsatellite loci. A grid was placed over each colony before spawning, and the emerging egg/sperm bundles were collected separately in each grid. The underlying tissues as well as the egg/sperm bundles were genotyped to determine whether different genotypes were present. Within the 14 mature colonies, we detected 10 colonies with more than one genotype (intracolonial genetic variability). Four out of these 10 mature colonies showed a transfer of different genotypes via the eggs to the next generation. In two out of these four cases, we found additional alleles, and in the two other cases, we found only a subset of alleles in the unfertilized eggs. Our results suggest that during reproduction of A. hyacinthus, more than one genotype per colony is able to reproduce. We discuss the occurrence of different genotypes within a single coral colony and the ability for those to release eggs which are genetically distinct.     

Diverse societies are more productive: a lesson from ants

The fitness consequences of animal personalities (also known as behavioural syndromes) have recently been studied in several solitary species. However, the adaptive significance of collective personalities in social insects and especially of behavioural variation among group members remains largely unexplored. Although intracolonial behavioural variation is an important component of division of labour, and as such a key feature for the success of societies, empirical links between behavioural variation and fitness are scarce. We investigated aggression, exploration and brood care behaviour in Temnothorax longispinosus ant colonies. We focused on two distinct aspects: intercolonial variability and its consistency across time and contexts, and intracolonial variability and its influence on productivity. Aggressiveness was consistent over four to five months with a new generation of workers emerging in between trial series. Other behaviours were not consistent over time. Exploration of novel environments responded to the sequence of assays: colonies were faster in discovering when workers previously encountered opponents in aggression experiments. Suites of correlated behaviours (e.g. aggression-exploration syndrome) present in the first series did not persist over time. Finally, colonies with more intracolonial behavioural variation in brood care and exploration of novel objects were more productive under standardized conditions than colonies with less variation.     

Multiple virus infections occur in individual polygyne and monogyne Solenopsis invicta ants

Concurrent infections of Solenopsis invicta colonies with S. invicta virus 1 (SINV-1), SINV-2, and SINV-3 has been reported. However, whether individual ants were capable of supporting multiple virus infections simultaneously was not known, nor whether the social form of the colony (polygyne or monogyne) had an influence on the occurrence of multiple infection rates in individual ants. S. invicta field populations were sampled sequentially to establish whether multiple virus infections co-occurred in individual worker ants. In addition, the intra-colony virus infection rates were compared in monogyne and polygyne field colonies to determine whether social form played a role in the viral infection prevalence. All combinations of virus infection (SINV-1, SINV-2, or SINV-3 alone, SINV-1 & SINV-2, SINV-1 & SINV-3, SINV-2 & SINV-3, and SINV-1, SINV-2 & SINV-3) were detected in individual worker ants as well as queens in the field. Thus, individual S. invicta ants can be infected simultaneously with all combinations of the S. invicta viruses. Colony social form did have an influence on the intra-colony prevalence of multiple S. invicta virus infections. Polygyne colonies exhibited significantly greater intra- and inter-colony single and multiple virus infections compared with monogyne colonies.     

A blend of formic acid,benzoic acid,and aliphatic alkanes mediates alarm recruitment responses in western carpenter ants,Camponotus modoc

Formicine ants in distress spray alarm pheromone which typically recruits nestmates for help. Studying the western carpenter ant, Camponotus modoc Wheeler (Hymenoptera: Formicidae), our objectives were to (1) determine the exocrine glands that contain alarm recruitment pheromone, (2) identify the key alarm recruitment pheromone components, and (3) ascertain the pheromone components that are discharged by distressed ants. In Y-tube olfactometer experiments, extracts of poison glands, but not of Dufour’s glands, elicited anemotactic responses from worker ants. Gas chromatographic-mass spectrometric analyses of poison gland extracts revealed the presence of (1) aliphatic alkanes (undecane, tridecane, pentadecane, heptadecane), (2) aliphatic alkenes [(Z)-7-pentadecene, (Z)-7- and (Z)-8-heptadecene], (3) two acids (formic, benzoic), and (4) other oxygenated compounds (hexadecan-1-ol, hexadecyl formate, hexadecyl acetate). Testing the responses of worker ants in Y-tube olfactometers to complete and partial synthetic blends of these compounds revealed that the acids and the alkanes are essential alarm pheromone components. In two-choice arena bioassays, micro-locations treated with synthetic alarm pheromone recruited worker ants. Acids and alkanes were abundant in the poison gland and the Dufour’s gland, respectively, suggesting that the alarm pheromone components originate from both glands. Moreover, alarm pheromone sprays of ants differed in that all sprays contained formic acid but only some also contained alkanes, implying that ants can independently discharge the content of either one or both glands in accordance with the type of distress incident they experience.     

Comparative morphology of cephalic exocrine glands among castes of the black ant Lasius niger

The glandular system is crucially involved in main aspects of ant social life. The function of glands has been primarily studied in the workers (the non-reproductive individuals in a colony). In contrast, little information is available on queens (the reproductive females in a colony) or males in spite of the obvious functional differences between these castes. Here we report a comparison of the general morphology of the mandibular, propharyngeal and postpharyngeal glands between the three castes of the black ant Lasius niger. The analysis clearly shows that all these cephalic glands differ in relative size between castes and suggests a link between gland structure and its behavioral role in queens, workers and males. In particular, males present a hypertrophied mandibular gland. This is consistent with the fact that these glands might be the source of the sex pheromone in this caste. By contrast, queens exhibited the most developed postpharyngeal glands. This is consistent with the production of particular cues by queens for workers to help them to distinguish between reproductive and non-reproductive females. Finally, the propharyngeal glands were most developed in the worker caste and of similar relative size in males and queens. Their function is still enigmatic.     

The Mediterranean bryozoan Myriapora truncata (Pallas, 1766): a potential indicator of (palaeo‐) environmental conditions

Fossil and Recent specimens of the Mediterranean bryozoan Myriapora truncata show considerable intra‐ and intercolonial differences in branch diameter and zooid size. Statistically significant variability occurs within colonies, between colonies within sites, and between sampled sites, while the presence of intracolonial variability clearly shows that branch diameter is largely controlled by environmental parameters. The three structural traits measured (branch diameter, zooid size and zooid depth) do not correlate, thus indicating a disconnection between the controls on overall zooid size and branch diameter. Possible environmental parameters that may have an influence on morphology are temperature, food supply or current energy. Whereas current energy has an effect on the colony branching pattern (branch spacing), there are indications that temperature may be the main, but not the only, parameter controlling zooid size, and it is suggested that food supply largely determines the branch diameter in M. truncata. However, the identification of the decisive factors and quantification of the relationships between environmental and morphological change is beyond the scope of this study. The results nevertheless show that, if the control factors of morphological variability can be ascertained in Recent M. truncata, this species may prove to be an indicator of environmental conditions and their change at different spatial and temporal scales in Cenozoic to Recent Mediterranean habitats.     

Source of alate excitant pheromones in the red imported fire antSolenopsis invicta (Hymenoptera: Formicidae)

At the onset of mating flights inSolenopsis invicta, workers swarm excitedly over the mound as alates prepare to fly. Previous studies demonstrated that this excitement is stimulated by the male and female alates. We investigated the glandular source(s) of pheromones produced by the alates that cause excitement. The only common female and male alate body part that elicited excitement when crushed was the head. Within the head, excised mandibular glands were found to be responsible for worker excitement. Fire ant workers are very sensitive to external stimuli and some excitement was elicited by crushed female gasters and male thoraces, but the response was never as significant as with crushed heads. Tests with summer and winter alates revealed similar results, except that gasters of winter female alates had a greater excitant effect than did gasters of summer female alates. This may be due to the production of attractant pheromones by the poison glands of overwintering female alates. We conclude that the mandibular gland is the source of alate excitant pheromones.     

The function of the post-pharyngeal glands of the red imported fire ant, Solenopsis invicta buren

Using radiolabelled triglycerides and fatty acids we have shown that these 2 lipids are absorbed into the haemolymph of the imported fire ant, Solenopsis invicta Buren from the post-pharyngeal gland. The post-pharyngeal glands attain their greatest weight and contain the highest hexane extractable lipid in establishing queens which have just reared the first minum brood. The lipid content of the digestive system is greatest in queens initiating a mating flight with the majority of the lipid contained in the crop. During colony establishment by the queen the lipid content of the crop moves forward, some is fed to the developing larvae and some moves into the post-pharyngeal gland. The hexane extractable lipid from the postpharyngeal gland of newly mated queens consists of hydrocarbons, sterols. tri-, di-, and monoglycerides and free fatty acids with a trace of wax esters.Excision of the post-pharyngeal glands from mated established queens causes no noticeable change in their behaviour or the behaviour they elicit from the remainder of the colony. However, post-pharyngeal glandectomized females lost weight and died in ca. 2 months although their crop and midgut contained food. The post-pharyngeal glands therefore appear to function in much the same way as a gastric caecum.     

Movement of honey bee (Apis mellifera L.) queen mandibular gland pheromone in populous and unpopulous colonies

The mode of intranest transfer of the honey bee queen mandibular gland pheromone complex (QMP) was investigated in unpopulous and populous, slightly congested colonies, using synthetic QMP containing tritiated 9-keto-2(E)-decenoic acid, one of the QMP components. Radiolabel was rapidly transported from the center to the peripheral regions of the nest, and in a manner consistent with worker to worker transport. Population size and congestion had no effect on the relative rates of movement from the center to the periphery of the nest or on the mean amounts of radiolabel on individual bees. However, a significantly smaller proportion of the workers in the populous colonies received detectable amounts of radiolabel than in the uncongested colonies, and workers carrying especially large amounts of radiolabel were less numerous in the crowded colonies. It is suggested that, at the stage of colony development that the colonies were in, population size has more of an effect on intranest pheromone transmission than does crowding. Interference with pheromone transfer may occur only at higher levels of congestion than were created, and nearer to the reproductive phase of colony development. An alternative hypothesis is that colony crowding does not significantly affect QMP transport and that the onset of reproductive queen rearing may be associated more with changes in worker thresholds of response to QMP.     

Mandibular glands of male European beewolves, Philanthus triangulum (Hymenoptera, Crabronidae)

Males of a solitary digger wasp, the European beewolf, Philanthus triangulum, possess large mandibular glands that have been reported to produce a scent marking pheromone. We analysed the morphology and ultrastructure of these glands using light microscopy as well as scanning and transmission electron microscopy. The paired glands are located laterally in the head and each side consists of a larger and a smaller part. Both parts possess a collecting duct each with distinct openings at the mandible base. However, the collecting duct of the larger part is additionally connected to the pharynx through a lateral extension. The collecting ducts are bordered by a monolayered epithelium lined with cuticle that exhibits conspicuous ramified protuberances. About 1400 acini consisting of class 3 gland cells surround the ducts and are connected to them through conducting canals. The main components in the cytoplasm of these gland cells are mitochondria, well-developed smooth endoplasmatic reticulum, and electron lucent vesicles suggesting a high secretory activity. The connection between the large gland parts and the pharynx suggests that the secretion of the mandibular glands might not only be delivered directly onto the mandibles but might also be transported to and stored in the postpharyngeal gland.     

Simple Reaching Is Not So Simple: Association Between Hand Use and Grip Preferences in Captive Chimpanzees

We assessed the relationship between grip preference and hand use in chimpanzees in 2 experiments. In experiment 1, we evaluated consistency in hand use and grip preference across 4 food types. The chimpanzees showed population-level right-handedness and there are significant positive associations for both hand and grip use across food types. In experiment 2, we assessed validity of hand use in relation to grip preference in 2 colonies of chimpanzees via the same methodology. Differences in hand preferences between colonies were associated with variation in the observed grip preferences. There was no evidence of rearing effects on handedness in either colony. We discuss the overall results in the context of the evolution of handedness in relation to increasing motor demands as manifest in variation on grasping behavior.