Eusocial Colonies Research Articles

Resource adaptation drives the size-complexity rule in termites.

The size-complexity rule posits that the evolution of larger cooperative groups should favour more division of labour. Examples include more cell types in larger multicellular organisms, and more polymorphic castes in larger eusocial colonies. However, a correlation between division of labour and group size may reflect a shared response of both traits to resource availability and/or profitability. Here, this possibility was addressed by investigating the evolution of sterile caste number (worker and soldier morphotypes) in termites, a major clade of eusocial insects in which the drivers of caste polymorphism are poorly understood. A novel dataset on 90 termite species was compiled from the published literature. The analysis showed that sterile caste number did increase markedly with colony size. However, after controlling for resource adaptations and phylogeny, there was no evidence for this relationship. Rather, sterile caste number increased with increasing nest-food separation and decreased with soil-feeding, through changes in worker (but not soldier) morphotype number. Further, colony size increased with nest-food separation, thus driving the false correlation between sterile caste number and colony size. These findings support adaptation to higher energy acquisition as key to the rise of complex insect societies, with larger size being a by-product.

Conflict and conflict resolution in the major transitions.

Conflict and conflict resolution have been argued to be fundamental to the major transitions in evolution. These were key events in life's history in which previously independently living individuals cooperatively formed a higher-level individual, such as a multicellular organism or eusocial colony. Conflict has its central role because, to proceed stably, the evolution of individuality in each major transition required within-individual conflict to be held in check. This review revisits the role of conflict and conflict resolution in the major transitions, addressing recent work arguing for a minor role. Inclusive fitness logic suggests that differences between the kin structures of clones and sexual families support the absence of conflict at the origin of multicellularity but, by contrast, suggest that key conflicts existed at the origin of eusociality. A principal example is conflict over replacing the founding queen (queen replacement). Following the origin of each transition, conflict remained important, because within-individual conflict potentially disrupts the attainment of maximal individuality (organismality) in the system. The conclusion is that conflict remains central to understanding the major transitions, essentially because conflict arises from differences in inclusive fitness optima while conflict resolution can help the system attain a high degree of coincidence of inclusive fitness interests.

Phylogeny of the carpenter bees (Apidae: Xylocopinae) highlights repeated evolution of sociality.

Many groups of animals have evolved social behaviours in different forms, from intimate familial associations to the complex eusocial colonies of some insects. The subfamily Xylocopinae, including carpenter bees and their relatives, is a diverse clade exhibiting a wide range of social behaviours, from solitary to obligate eusociality with distinct morphological castes, making them ideal focal taxa in studying the evolution of sociality. We used ultraconserved element data to generate a broadly sampled phylogeny of the Xylocopinae, including several newly sequenced species. We then conducted ancestral state reconstructions on the evolutionary history of sociality in this group under multiple coding models. Our results indicate solitary origins for the Xylocopinae with multiple transitions to sociality across the tree and subsequent reversals to solitary life, demonstrating the lability and dynamic nature of social evolution in carpenter bees. Ultimately, this work clarifies the evolutionary history of the Xylocopinae, and expands our understanding of independent origins and gains and losses of social complexity.

Neotropical bee microbiomes point to a fragmented social core and strong species-level effects

BackgroundIndividuals that band together create new ecological opportunities for microorganisms. In vertical transmission, theory predicts a conserved microbiota within lineages, especially social bees. Bees exhibit solitary to social behavior among and/or within species, while life cycles can be annual or perennial. Bee nests may be used over generations or only once, and foraging ecology varies widely. To assess which traits are associated with bee microbiomes, we analyzed microbial diversity within solitary and social bees of Apidae, Colletidae, and Halictidae, three bee families in Panama’s tropical forests. Our analysis considered the microbiome of adult gut contents replicated through time, localities, and seasons (wet and dry) and included bee morphology and comparison to abdominal (dissected) microbiota. Diversity and distribution of tropical bee microbes (TBM) within the corbiculate bee clade were emphasized.ResultsWe found the eusocial corbiculate bees tended to possess a more conserved gut microbiome, attributable to vertical transmission, but microbial composition varied among closely related species. Euglossine bees (or orchid bees), corbiculates with mainly solitary behavior, had more variable gut microbiomes. Their shorter-tongued and highly seasonal species displayed greater diversity, attributable to flower-visiting habits. Surprisingly, many stingless bees, the oldest corbiculate clade, lacked bacterial genera thought to predate eusociality, while several facultatively social, and solitary bee species possessed those bacterial taxa. Indeed, nearly all bee species displayed a range of affinities for single or multiple variants of the “socially associated” bacterial taxa, which unexpectedly demonstrated high sequence variation.ConclusionsTaken together, these results call into question whether specific bacterial associates facilitate eusocial behavior, or are subsequently adopted, or indicate frequent horizontal transmission between perennial eusocial colonies and other social, facultatively social, and solitary bees.7ptKwZS8umiRRciGf9GNsXVideo

Biogeography and evolution of social parasitism in Australian Myrmecia bulldog ants revealed by phylogenomics

Studying the historical biogeography and life history transitions from eusocial colony life to social parasitism contributes to our understanding of the evolutionary mechanisms generating biodiversity in eusocial insects. The ants in the genus Myrmecia are a well-suited system for testing evolutionary hypotheses about how their species diversity was assembled through time because the genus is endemic to Australia with the single exception of the species M. apicalis inhabiting the Pacific Island of New Caledonia, and because at least one social parasite species exists in the genus. However, the evolutionary mechanisms underlying the disjunct biogeographic distribution of M. apicalis and the life history transition(s) to social parasitism remain unexplored. To study the biogeographic origin of the isolated, oceanic species M. apicalis and to reveal the origin and evolution of social parasitism in the genus, we reconstructed a comprehensive phylogeny of the ant subfamily Myrmeciinae. We utilized Ultra Conserved Elements (UCEs) as molecular markers to generate a molecular genetic dataset consisting of 2,287 loci per taxon on average for 66 out of the 93 known Myrmecia species as well as for the sister lineage Nothomyrmecia macrops and selected outgroups. Our time-calibrated phylogeny inferred that: (i) stem Myrmeciinae originated during the Paleocene ∼ 58 Ma ago; (ii) the current disjunct biogeographic distribution of M. apicalis was driven by long-distance dispersal from Australia to New Caledonia during the Miocene ∼ 14 Ma ago; (iii) the single social parasite species, M. inquilina, evolved directly from one of the two known host species, M. nigriceps, in sympatry via the intraspecific route of social parasite evolution; and (iv) 5 of the 9 previously established taxonomic species groups are non-monophyletic. We suggest minor changes to reconcile the molecular phylogenetic results with the taxonomic classification. Our study enhances our understanding of the evolution and biogeography of Australian bulldog ants, contributes to our knowledge about the evolution of social parasitism in ants, and provides a solid phylogenetic foundation for future inquiries into the biology, taxonomy, and classification of Myrmeciinae.

Reconstructing queen genotypes by pool sequencing colonies in eusocial insects: Statistical Methods and their application to honeybee.

Eusocial insects are crucial to many ecosystems, and particularly the honeybee (Apis mellifera). One approach to facilitate their study in molecular genetics, is to consider whole-colony genotyping by combining DNA of multiple individuals in a single pool sequencing experiment. Cheap and fast, this technique comes with the drawback of producing data requiring dedicated methods to be fully exploited. Despite this limitation, pool sequencing data have been shown to be informative and cost-effective when working on random mating populations. Here, we present new statistical methods for exploiting pool sequencing of eusocial colonies in order to reconstruct the genotypes of the queen of such colony. This leverages the possibility to monitor genetic diversity, perform genomic-based studies or implement selective breeding. Using simulations and honeybee real data, we show that the new methods allow for a fast and accurate estimation of the queen's genetic ancestry, with correlations of about 0.9 to that obtained from individual genotyping. Also, it allows for an accurate reconstruction of the queen genotypes, with about 2% genotyping error. We further validate these inferences using experimental data on colonies with both pool sequencing and individual genotyping of drones. In brief, in this study we present statistical models to accurately estimate the genetic ancestry and reconstruct the genotypes of the queen from pool sequencing data from workers of an eusocial colony. Such information allows to exploit pool sequencing for traditional population genetics analyses, association studies and for selective breeding. While validated in Apis mellifera, these methods are applicable to other eusocial hymenopterans.

Exploring Evolved Multicellular Life Histories in a Open-Ended Digital Evolution System

Evolutionary transitions occur when previously-independent replicating entities unite to form more complex individuals. Such transitions have profoundly shaped natural evolutionary history and occur in two forms: fraternal transitions involve lower-level entities that are kin (e.g., transitions to multicellularity or to eusocial colonies), while egalitarian transitions involve unrelated individuals (e.g., the origins of mitochondria). The necessary conditions and evolutionary mechanisms for these transitions to arise continue to be fruitful targets of scientific interest. Here, we examine a range of fraternal transitions in populations of open-ended self-replicating computer programs. These digital cells were allowed to form and replicate kin groups by selectively adjoining or expelling daughter cells. The capability to recognize kin-group membership enabled preferential communication and cooperation between cells. We repeatedly observed group-level traits that are characteristic of a fraternal transition. These included reproductive division of labor, resource sharing within kin groups, resource investment in offspring groups, asymmetrical behaviors mediated by messaging, morphological patterning, and adaptive apoptosis. We report eight case studies from replicates where transitions occurred and explore the diverse range of adaptive evolved multicellular strategies.

Phenological and social characterization of three Lasioglossum (Dialictus) species inferred from long-term trapping collections

Detailed social and phenological data collected from nesting aggregations exist for relatively few sweat bee species because nesting aggregations are rarely found in large numbers, even when local populations are highly abundant. This limits researchers’ abilities to assess the social status of many species, which in turn, limits our ability to trace the sequence of evolutionary steps between alternative social states. To address this problem, we demonstrate the utility of rehydrated, pinned specimens from pan trap and netting collections for generating inferences about the phenology and social status of a well-studied sweat bee species, Lasioglossum (Dialictus) laevissimum. A detailed comparison of phenology and reproductive traits, between pinned specimens and those in a previous nesting study, produced similar results for bivoltine foraging activity and eusocial colony organization typical in this species. We then used pinned specimens from monitoring studies to describe, for the first time, the foraging phenology and social behaviour of two additionalDialictusspecies,L. hitchensiandL. ellisiae. BothL. hitchensiandL. ellisiaeeach exhibited two peaks in abundance during their breeding seasons, indicating two periods of foraging activity, which correspond to provisioning of spring and summer broods. Differences in body size, wear, and ovarian development of spring and summer females indicated thatL. hitchensiis most likely eusocial, whileL. ellisiaeis either solitary or communal. This study demonstrates that analyses of specimens obtained from flower and pan trap collections can be used for assessing the phenology and social organization of temperate sweat bees in the absence of nesting data. The phenological and social lability of many sweat bee species make them ideal for studying geographic and temporal variability in sociality, and analyses of pan trap collections can make these studies possible when direct observations are impossible.

Hybridization enables the fixation of selfish queen genotypes in eusocial colonies.

A eusocial colony typically consists of two main castes: queens that reproduce and sterile workers that help them. This division of labor, however, is vulnerable to genetic elements that favor the development of their carriers into queens. Several factors, such as intracolonial relatedness, can modulate the spread of such caste‐biasing genotypes. Here we investigate the effects of a notable yet understudied ecological setting: where larvae produced by hybridization develop into sterile workers. Using mathematical modeling, we show that the coevolution of hybridization with caste determination readily triggers an evolutionary arms race between nonhybrid larvae that increasingly develop into queens, and queens that increasingly hybridize to produce workers. Even where hybridization reduces worker function and colony fitness, this race can lead to the loss of developmental plasticity and to genetically hard‐wired caste determination. Overall, our results may help understand the repeated evolution toward remarkable reproductive systems (e.g., social hybridogenesis) observed in several ant species.

Long bone histomorphogenesis of the naked mole-rat: Histodiversity and intraspecific variation.

Lacking fur, living in eusocial colonies and having the longest lifespan of any rodent, makes naked mole-rats (NMRs) rather peculiar mammals. Although they exhibit a high degree of polymorphism, skeletal plasticity and are considered a novel model to assess the effects of delayed puberty on the skeletal system, scarce information on their morphogenesis exists. Here, we examined a large ontogenetic sample (n=76) of subordinate individuals to assess the pattern of bone growth and bone microstructure of fore- and hindlimb bones by using histomorphological techniques. Over 290 undecalcified thin cross-sections from the midshaft of the humerus, ulna, femur, and tibia from pups, juveniles and adults were analyzed with polarized light microscopy. Similar to other fossorial mammals, NMRs exhibited a systematic cortical thickening of their long bones, which clearly indicates a conserved functional adaptation to withstand the mechanical strains imposed during digging, regardless of their chisel-tooth predominance. We describe a high histodiversity of bone matrices and the formation of secondary osteons in NMRs. The bones of pups are extremely thin-walled and grow by periosteal bone formation coupled with considerable expansion of the medullary cavity, a process probably tightly regulated and adapted to optimize the amount of minerals destined for skeletal development, to thus allow the female breeder to produce a higher number of pups, as well as several litters. Subsequent cortical thickening in juveniles involves high amounts of endosteal bone apposition, which contrasts with the bone modeling of other mammals where a periosteal predominance exists. Adults have bone matrices predominantly consisting of parallel-fibered bone and lamellar bone, which indicate intermediate to slow rates of osteogenesis, as well as the development of poorly vascularized lamellar-zonal tissues separated by lines of arrested growth (LAGs) and annuli. These features reflect the low metabolism, low body temperature and slow growth rates reported for this species, as well as indicate a cyclical pattern of osteogenesis. The presence of LAGs in captive individuals was striking and indicates that postnatal osteogenesis and its consequent cortical stratification most likely represents a plesiomorphic thermometabolic strategy among endotherms which has been suggested to be regulated by endogenous rhythms. However, the generalized presence of LAGs in this and other subterranean taxa in the wild, as well as recent investigations on variability of environmental conditions in burrow systems, supports the hypothesis that underground environments experience seasonal fluctuations that may influence the postnatal osteogenesis of animals by limiting the extension of burrow systems during the unfavorable dry seasons and therefore the finding of food resources. Additionally, the intraspecific variation found in the formation of bone tissue matrices and vascularization suggested a high degree of developmental plasticity in NMRs, which may help explaining the polymorphism reported for this species. The results obtained here represent a valuable contribution to understanding the relationship of several aspects involved in the morphogenesis of the skeletal system of a mammal with extraordinary adaptations.

Split sex ratios and genetic relatedness in a primitively eusocial sweat bee

In eusocial Hymenoptera, queens and their helper offspring should favour different sex investment ratios. Queens should prefer a 1:1 investment ratio, as they are equally related to offspring of both sexes (r = 0.5). In contrast, helpers should favour an investment ratio of 3:1 towards the production of female brood. This conflict arises because helpers are more closely related to full sisters (r = 0.75) than brothers (r = 0.25). However, helpers should invest relatively more in male brood if relatedness asymmetry within their colony is reduced. This can occur due to queen replacement after colony orphaning, multiple paternity and the presence of unrelated alien helpers. We analysed an unprecedentedly large number of colonies (n = 109) from a UK population of Lasioglossum malachurum, an obligate eusocial sweat bee, to tease apart the effects of these factors on colony-level investment ratios. We found that multiple paternity, unrelated alien helpers and colony orphaning were all common. Queen-right colonies invested relatively more in females than did orphaned colonies, producing a split sex ratio. However, investment ratios did not change due to multiple paternity or the presence of alien helpers, reducing inclusive fitness pay-offs for helpers. Queen control may also have been important: helpers rarely laid male eggs, and investment in female brood was lower when queens were large relative to their helpers. Genetic relatedness between helpers and the brood that they rear was 0.43 in one year and 0.37 in another year, suggesting that ecological benefits, as well as relatedness benefits, are necessary for the maintenance of helping behaviour.Significance statementHow helping behaviour is maintained in eusocial species is a key topic in evolutionary biology. Colony-level sex investment ratio changes in response to relatedness asymmetries can dramatically influence inclusive fitness benefits for helpers in eusocial Hymenoptera. The extent to which helpers in primitively eusocial colonies can respond adaptively to different sources of variation in relatedness asymmetry is unclear. Using data from 109 colonies of the sweat bee Lasioglossum malachurum, we found that queen loss, but not multiple paternity or the presence of alien helpers, was correlated with colony sex investment ratios. Moreover, we quantified average helper-brood genetic relatedness to test whether it is higher than that predicted under solitary reproduction (r = 0.5). Values equal to and below r = 0.5 suggest that relatedness benefits alone cannot explain the maintenance of helping behaviour. Ecological benefits of group living and/or coercion must also contribute.

Characterization of brown adipose tissue thermogenesis in the naked mole-rat (Heterocephalus glaber), a heterothermic mammal

The naked mole-rat (NMR) is a heterothermic mammal that forms eusocial colonies consisting of one reproductive female (queen), several reproductive males, and subordinates. Despite their heterothermy, NMRs possess brown adipose tissue (BAT), which generally induces thermogenesis in cold and some non-cold environments. Previous studies suggest that NMR-BAT induces thermogenesis by cold exposure. However, detailed NMR-BAT characteristics and whether NMR-BAT thermogenesis occurs in non-cold environments are unknown. Here, we show beta-3 adrenergic receptor (ADRB3)-dependent thermogenic potential of NMR-BAT, which contributes to thermogenesis in the isolated queen in non-cold environments (30 °C). NMR-BAT expressed several brown adipocyte marker genes and showed noradrenaline-dependent thermogenic activity in vitro and in vivo. Although our ADRB3 inhibition experiments revealed that NMR-BAT thermogenesis slightly delays the decrease in body temperature in a cold environment (20 °C), it was insufficient to prevent the decrease in the body temperatures. Even at 30 °C, NMRs are known to prevent the decrease of and maintain their body temperature by heat-sharing behaviors within the colony. However, isolated NMRs maintained their body temperature at the same level as when they are in the colony. Interestingly, we found that queens, but not subordinates, induce BAT thermogenesis in this condition. Our research provides novel insights into NMR thermoregulation.

Reproductive physiology corresponds to adult nutrition and task performance in a Neotropical paper wasp: a test of dominance-nutrition hypothesis predictions

The dominance-nutrition hypothesis predicts that nutritional intake and energetic costs in adulthood interact to drive behavioral and physiological differences between females in primitively eusocial insects, and thereby affect reproductive caste. We tested predictions of this hypothesis in independent-founding Mischocyttarus pallidipectus paper wasps. We measured stable isotope tissue composition to compare nutritional status before adult emergence and among adult females. Adult tissue δ15N content (an indicator of feeding at a higher trophic level because it is enriched in animal prey relative to plant-based foods) was significantly higher in adults than at the end of pupal development, suggesting adult nutrition affects δ15N content after the end of pupal development. We then asked if behavior and nutritional status predicted ovary development. We measured ovary development, nitrogen and carbon stable isotope ratios, dominance behavior, and task performance (foraging, as indicated by time spent on the nest) for adult female wasps. We used social network analysis to quantify differences in social status between females with developed and undeveloped (filamentous) ovaries. Dominant females spent more time on the nest and were significantly more enriched in δ15N than subordinate females. These data support the dominance-nutrition hypothesis: adult behavior and energy expenditure, and access to animal-based diets, correspond to female reproductive physiology, and may play a role in adult-stage caste determination. Within animal social groups, differential access to resources can affect differences in reproductive success. This may affect caste determination in eusocial colonies, where reproductive tasks are divided between female castes, with a queen (or multiple queens) capable of egg-laying and helped by sterile workers. Female sterile workers and reproductives of some social Hymenoptera are morphologically similar at adult emergence, suggesting that adult experience may impact caste status. We asked whether patterns of nutritional physiology in Mischocyttarus pallidipectus paper wasps matched adult-stage caste determination by testing predictions of the dominance-nutrition hypothesis, which states that adult nutrition and energetic costs of tasks shape reproductive female development through behavioral and physiological changes.

The hypoxia tolerance of eight related African mole-rat species rivals that of naked mole-rats, despite divergent ventilatory and metabolic strategies in severe hypoxia.

Burrowing mammals tend to be more hypoxia tolerant than non-burrowing mammals and rely less on increases in ventilation and more on decreases in metabolic rate to tolerate hypoxia. Naked mole-rats (Heterocephalus glaber, NMRs), eusocial mammals that live in large colonies, are among the most hypoxia-tolerant mammals, and rely almost solely on decreases in metabolism with little change in ventilation during hypoxia. We hypothesized that the remarkable hypoxia toleranceof NMRs is an evolutionarily conserved trait derived from repeated exposure to severe hypoxia owing to their burrow environment and eusocial colony organization. We used whole-body plethysmography and indirect calorimetry to measure the hypoxic ventilatory and metabolic responses of eight mole-rat species closely related to the NMR. We found that all eightspecies examined had a strong tolerance to hypoxia, with most species tolerating 3kPa O2 , Heliophobius emini tolerating 2kPa O2 and Bathyergus suillus tolerating 5kPa O2 . All species examined employed a combination of increases in ventilation and decreases in metabolism in hypoxia, a response midway between that of the NMR and that of other fossorial species (larger ventilatory responses, lesser reductions in metabolism). We found that eusociality is not fundamental to the physiological response to hypoxia of NMRs as Fukomys damarensis, another eusocial species, was among this group. Our data suggest that, while the NMR is unique in the pattern of their physiological response to hypoxia, eight closely related mole-rat species share the ability to tolerate hypoxia like the current "hypoxia-tolerant champion," the NMR.

The Better to Eat You With: Bite Force in the Naked Mole-Rat (Heterocephalus glaber) Is Stronger Than Predicted Based on Body Size.

Naked mole-rats (Heterocephalus glaber) are subterranean rodents that utilize their incisors for feeding, chisel-tooth digging of complex tunnel systems, social interactions, and defense in their eusocial colony structure. Previous studies have shown that naked mole-rats have morphological and anatomical adaptations that predict strong bite forces, namely, skulls that are relatively tall and wide, in addition to impressive masticatory musculature. However, no studies to date have directly measured bite force in this species or analyzed the relationship between bite force and social caste. In the current study, we assessed adult naked mole-rat maximum bite force in relation to body mass, in addition to considering each animal’s position within the eusocial hierarchy (i.e., dominant versus subordinate). Each animal was permitted to freely interact with a piezo-resistive bite force sensor. Our results showed that bite force was correlated with body mass in subordinate but not in dominant naked mole-rats, and that subordinate animals exhibited a shorter latency in producing their first bite. Maximum bite force was significantly influenced by caste. In comparing bite force with available data from previous studies across 82 additional mammalian species, subordinate naked mole-rats exhibited a bite force that was 65% higher than predicted for their body size, comparable to Tasmanian devils and exceeding bite force values for all of the carnivorans included for comparison. These results supported the hypothesis that the naked mole-rat’s bite force would exceed predictions based on body size due to the behavioral importance and specialization of the naked mole-rat incisors. This study provides insight into the differences in bite force across species, and the significant role that social and ecological factors might play in the evolutionary relationship between bite force performance and underlying anatomical structures.

Revising the Superorganism: An Organizational Approach to Complex Eusociality.

Eusociality is broadly defined as: colonies consisting of overlapping generations, cooperative brood care, and a reproductive division of labor where sterile (or non-reproductive) workers help the reproductive members. Colonies of many complex eusocial insect species (e.g., ants, bees, termites) exhibit traits, at the collective level, that are more analogous to biological individuals rather than to groups. Indeed, due to this, colonies of the most complex species are typically a unit of selection, which has led many authors to once again apply the concept of the superorganism to eusocial insects. However, unlike Wheeler, who originally employed the concept from a physiological and evolutionary perspective, today the superorganism is typically understood only from an evolutionary perspective, using group selection. This is because of the widely held view that eusocial colonies are self-organized systems. According to this view, even the most complex eusocial systems can be explained by appealing to a set of local interactions between parts of an initially disordered system (i.e., self-organization), without the need of any hierarchical control. In this paper, we challenge the mainstream view that hierarchical control and regulation does not occur, or is not necessary, in complex eusocial colonies. Using a case study of honey bees (Apis mellifera), we develop an alternative to the self-organization approach that focuses on the hierarchical nature of the organization of complex eusocial systems—that we refer to as the hierarchical-organizational approach. In addition, we analyze how colonies of eusocial insects show a complex set of interactions between the different organisms that bring forth a new cohesive collective organization, and how in turn the constitutive entities of this collective organization are transformed in this process. This paper argues that an inter-identity (namely the superorganism) emerges at the collective level in complex eusocial colonies, such as honey bees, due to the hierarchically organized network of interactions within the colony.

Queen Recognition Signals in Two Primitively Eusocial Halictid Bees: Evolutionary Conservation and Caste-Specific Perception.

Queen signals are known to regulate reproductive harmony within eusocial colonies by influencing worker behavior and ovarian physiology. However, decades of research have resulted in the identification of just a few queen signals, and studies of their mode of action are rare. Our aim was to identify queen recognition signals in the halictid bee Lasioglossum pauxillum and to analyze caste differences in the olfactory perception of queen signals in L. pauxillum and the closely related species L. malachurum. We performed chemical analyses and bioassays to test for caste differences in chemical profiles and worker behavior influenced by queen-specific compounds in L. pauxillum. Our results indicated that caste differences in the chemical profiles were mainly attributable to higher amounts of macrocyclic lactones in queens. Bioassays demonstrated a higher frequency of subordinate behavior in workers elicited by queen-specific amounts of macrocyclic lactones. Thus, macrocyclic lactones function as queen recognition signals in L. pauxillum, as in L. malachurum. Using electrophysiological analyses, we have demonstrated that queens of both tested species lack antennal reactions to certain macrocyclic lactones. Therefore, we assume that this is a mechanism to prevent reproductive self-inhibition in queens. Our results should stimulate debate on the conservation and mode of action of queen signals.

Testicular Structure and Spermatogenesis in the Naked Mole-Rat Is Unique (Degenerate) and Atypical Compared to Other Mammals.

The naked mole-rat (NMR) queen controls reproduction in her eusocial colony by usually selecting one male for reproduction and suppressing gametogenesis in all other males and females. Simplified, polymorphic and slow-swimming spermatozoa in the NMR seem to have been shaped by a low risk of sperm competition. We hypothesize that this unique mammalian social organization has had a dramatic influence on testicular features and spermatogenesis in the NMR. The testicular structure as well as spermatogenic cell types and its organization in breeding, subordinate and disperser males were studied using light microscopy and transmission electron microscopy. Even though the basic testicular design in NMRs is similar to most Afrotheria as well as some rodents with intra-abdominal testes, the Sertoli and spermatogenic cells have many atypical mammalian features. Seminiferous tubules are distended and contain large volumes of fluid while interstitial tissue cover about 50% of the testicular surface area and is mainly composed of Leydig cells. The Sertoli cell cytoplasm contains an extensive network of membranes and a variety of fluid-containing vesicles. Furthermore, Sertoli cells form numerous phagosomes that often appear as extensive accumulations of myelin. Another unusual feature of mature NMR Sertoli cells is mitotic division. While the main types of spermatogonia and spermatocytes are clearly identifiable, these cells are poorly organized and many spermatids without typical intercellular bridges are present. Spermatid heads appear to be malformed with disorganized chromatin, nuclear fragmentation and an ill-defined acrosome formed from star-like Golgi bodies. Rudimentary manchette development corresponds with the occurrence of abnormal sperm morphology. We also hypothesize that NMR testicular organization and spermiation are modified to produce spermatozoa on demand in a short period of time and subsequently use a Sertoli cell “pump” to flush the spermatozoa into short tubuli recti and simplified rete testis. Despite the difficulty in finding cellular associations during spermatogenesis, six spermatogenic stages could be described in the NMR. These numerous atypical and often simplified features of the NMR further supports the notion of degenerative orthogenesis that was selected for due to the absence of sperm competition.

Sex-allocation conflict and sexual selection throughout the lifespan of eusocial colonies.

Models of sex‐allocation conflict are central to evolutionary biology but have mostly assumed static decisions, where resource allocation strategies are constant over colony lifespan. Here, we develop a model to study how the evolution of dynamic resource allocation strategies is affected by the queen‐worker conflict in annual eusocial insects. We demonstrate that the time of dispersal of sexuals affects the sex‐allocation ratio through sexual selection on males. Furthermore, our model provides three predictions that depart from established results of classic static allocation models. First, we find that the queen wins the sex‐allocation conflict, while the workers determine the maximum colony size and colony productivity. Second, male‐biased sex allocation and protandry evolve if sexuals disperse directly after eclosion. Third, when workers are more related to new queens, then the proportional investment into queens is expected to be lower, which results from the interacting effect of sexual selection (selecting for protandry) and sex‐allocation conflict (selecting for earlier switch to producing sexuals). Overall, we find that colony ontogeny crucially affects the outcome of sex‐allocation conflict because of the evolution of distinct colony growth phases, which decouples how queens and workers affect allocation decisions and can result in asymmetric control.

Responses to pup vocalizations in subordinate naked mole-rats are induced by estradiol ingested through coprophagy of queen’s feces

Naked mole-rats form eusocial colonies consisting of a single breeding female (the queen), several breeding males, and sexually immature adults (subordinates). Subordinates are cooperative and provide alloparental care by huddling and retrieving pups to the nest. However, the physiological mechanism(s) underlying alloparental behavior of nonbreeders remains undetermined. Here, we examined the response of subordinates to pup voice and the fecal estradiol concentrations of subordinates during the three reproductive periods of the queen, including gestation, postpartum, and nonlactating. Subordinate response to pup voice was observed only during the queen's postpartum and was preceded by an incremental rise in subordinates' fecal estradiol concentrations during the queen's gestation period, which coincided with physiological changes in the queen. We hypothesized that the increased estradiol in the queen's feces was disseminated to subordinates through coprophagy, which stimulated subordinates' responses to pup vocalizations. To test this hypothesis, we fed subordinates either fecal pellets from pregnant queens or pellets from nonpregnant queens amended with estradiol for 9 days and examined their response to recorded pup voice. In both treatments, the subordinates exhibited a constant level of response to pup voice during the feeding period but became more responsive 4 days after the feeding period. Thus, we believe that we have identified a previously unknown system of communication in naked mole-rats, in which a hormone released by one individual controls the behavior of another individual and influences the level of responsiveness among subordinate adults to pup vocal signals, thereby contributing to the alloparental pup care by subordinates.