Eusocial Hymenopterans Research Articles

Alternative double strand break repair pathways shape the evolution of high recombination in the honey bee, Apis mellifera.

Social insects, particularly honey bees, have exceptionally high genomic frequencies of genetic recombination. This phenomenon and underlying mechanisms are poorly understood. To characterise the patterns of crossovers and gene conversion in the honey bee genome, a recombination map of 187 honey bee brothers was generated by whole-genome resequencing. Recombination events were heterogeneously distributed without many true hotspots. The tract lengths between phase shifts were bimodally distributed, indicating distinct crossover and gene conversion events. While crossovers predominantly occurred in G/C-rich regions and seemed to cause G/C enrichment, the gene conversions were found predominantly in A/T-rich regions. The nucleotide composition of sequences involved in gene conversions that were associated with or distant from crossovers corresponded to the differences between crossovers and gene conversions. These combined results suggest two types of DNA double-strand break repair during honey bee meiosis: non-canonical homologous recombination, leading to gene conversion and A/T enrichment of the genome, and the canonical homologous recombination based on completed double Holliday Junctions, which can result in gene conversion or crossover and is associated with G/C bias. This G/C bias may be selected for to balance the A/T-rich base composition of eusocial hymenopteran genomes. The lack of evidence for a preference of the canonical homologous recombination for double-strand break repair suggests that the high genomic recombination rate of honey bees is mainly the consequence of a high rate of double-strand breaks, which could in turn result from the life history of honey bees and their A/T-rich genome.

Adaptive evolution of A-to-I auto-editing site in Adar of eusocial insects

BackgroundAdenosine-to-inosine (A-to-I) RNA editing is a co-/post-transcriptional modification introducing A-to-G variations in RNAs. There is extensive discussion on whether the flexibility of RNA editing exerts a proteomic diversification role, or it just acts like hardwired mutations to correct the genomic allele. Eusocial insects evolved the ability to generate phenotypically differentiated individuals with the same genome, indicating the involvement of epigenetic/transcriptomic regulation.MethodsWe obtained the genomes of 104 Hymenoptera insects and the transcriptomes of representative species. Comparative genomic analysis was performed to parse the evolutionary trajectory of a regulatory Ile > Met auto-recoding site in Adar gene.ResultsAt genome level, the pre-editing Ile codon is conserved across a node containing all eusocial hymenopterans. At RNA level, the editing events are confirmed in representative species and shows considerable condition-specificity. Compared to random expectation, the editable Ile codon avoids genomic substitutions to Met or to uneditable Ile codons, but does not avoid mutations to other unrelated amino acids.ConclusionsThe flexibility of Adar auto-recoding site in Hymenoptera is selectively maintained, supporting the flexible RNA editing hypothesis. We proposed a new angle to view the adaptation of RNA editing, providing another layer to explain the great phenotypical plasticity of eusocial insects.

Pervasive relaxed selection in termite genomes.

Genetic changes that enabled the evolution of eusociality have long captivated biologists. More recently, attention has focussed on the consequences of eusociality on genome evolution. Studies have reported higher molecular evolutionary rates in eusocial hymenopteran insects compared with their solitary relatives. To investigate the genomic consequences of eusociality in termites, we analysed nine genomes, including newly sequenced genomes from three non-eusocial cockroaches. Using a phylogenomic approach, we found that termite genomes have experienced lower rates of synonymous substitutions than those of cockroaches, possibly as a result of longer generation times. We identified higher rates of non-synonymous substitutions in termite genomes than in cockroach genomes, and identified pervasive relaxed selection in the former (24-31% of the genes analysed) compared with the latter (2-4%). We infer that this is due to reductions in effective population size, rather than gene-specific effects (e.g. indirect selection of caste-biased genes). We found no obvious signature of increased genetic load in termites, and postulate efficient purging of deleterious alleles at the colony level. Additionally, we identified genomic adaptations that may underpin caste differentiation, such as genes involved in post-translational modifications. Our results provide insights into the evolution of termites and the genomic consequences of eusociality more broadly.

Accessory gland size increases with sperm competition intensity in Cataglyphis desert ants

In many species, females have multiple mates, whose sperm compete for paternity. Males may subsequently invest in the increased production of sperm and/or seminal fluid. The latter is a complex mixture of proteins, peptides, and other compounds generated by the accessory glands (AGs) and is transferred to females along with a male’s sperm. Seminal fluid is known to be a key determinant of competitive outcomes among sperm, and its production may trade off with that of sperm. We show that AG size—a proxy for seminal fluid production—has a positive and phylogenetically robust correlation with both sperm competition intensity and sperm production in nine species of Cataglyphis desert ants. These results indicate a lack of trade-off between sperm production and seminal fluid production. They underscore that sperm competition may strongly shape sperm traits and could drive reproductive performance in eusocial hymenopterans.

Flight capabilities of invasive yellowjacket Vespula germanica drones: the effect of kinship and nutrition

AbstractDispersal is a multifactorial process which may influence, among several others, population dynamics, inbreeding likelihood, and mate competition. For species that disperse by flight, many studies have suggested that life‐history traits, such as body size or nutritional condition, could promote variation in flight capabilities and, consequently, on dispersal behavior. Eusocial hymenopterans are likely to experience high densities of related reproductive individuals near their nests, thus increasing probabilities of inbred matings and kin competition. Spatial segregation resulting from differences in flight capabilities of Vespula germanica (Fabricius) (Hymenoptera: Vespidae) nestmate drones could reduce interactions with kin, thus decrease inbreeding and kin competition. Our aim was to characterize, in the laboratory, flight capabilities of sibling drones and elucidate traits that could affect it, focusing on the relevance of carbohydrate intake (diets with different sugar concentration) during early adulthood. Our results indicate that in tethered flight mill assays, V. germanica drones have highly variable flight capabilities within and between nests, with most drones likely to remain relatively close to their nest of origin. The observed flight capabilities are affected by nutrition, with those individuals better nurtured (i.e., with higher nutritional index), flying farther distances. Additionally, diets with variable carbohydrate concentration provided during early adulthood did not affect flight capabilities (only those exposed to a carbohydrate‐free diet had reduced flight distances and speeds), suggesting that nutrition during larval stages is determinant for flight. Our findings contribute to the understanding of the mechanisms underlying V. germanica drone dispersal behavior which, in turn, might influence inbreeding likelihood in the species.

Aversive reinforcement improves visual discrimination learning in free-flying wasps (Vespula vulgaris)

Understanding and assessing the capacity for learning, memory, and cognition in non-model organisms is a growing field. In invertebrate cognition, eusocial hymenopteran species such as honeybees, bumblebees, and ants are well-studied for their learning and memory abilities due to decades of research providing well-tested methods of training and assessing cognition. In the current study, we assess the use of different conditioning methods on visual learning in a non-model hymenopteran species which is becoming increasingly used in learning and memory tasks, the European wasp (Vespula vulgaris). We trained individual wasps to learn to discriminate between perceptually similar colours using absolute conditioning (reward on target stimulus in the absence of distractors), appetitive differential conditioning (reward on target stimulus and no outcome for incorrect stimulus), or appetitive-aversive differential conditioning (reward on target stimulus and aversive outcome for incorrect stimulus). When trained with absolute conditioning, wasps were unable to learn to discriminate between perceptually similar colours. However, when trained with appetitive differential conditioning or appetitive-aversive differential conditioning, wasps were able to learn to discriminate between two similar colours, although they performed best when an aversive reinforcement was provided during training. Our results show similarities to learning behaviour in honeybees and bumblebees, and provide insight into the learning and cognition of a non-model invertebrate. Our findings provide important comparative data to aid in understanding the evolution of learning and memory in hymenopterans.Significance statementForaging insects such as bees, ants, and wasps visit a variety of food sources such as flowers, insect prey, and rotting fruit. Many of these insects must learn and recall resource traits such as location, scent, shape, colour, and size. To understand the process of learning in insects which are comparatively less studied than those such as honeybees and bumblebees, we trained wasps to discriminate between two similar colours using three different types of conditioning methods. Wasps were best able to learn to discriminate between the similar colours when trained with appetitive-aversive differential conditioning, where a reward is provided for a correct choice and an aversive outcome was providing for an incorrect choice. Our results show that similar to popular invertebrate cognition models, wasp learning is improved when an aversive outcome is introduced for incorrect choices.

Age‐related changes in male ant sperm quality

AbstractSperm cells age during storage in the male reproductive organs, which can lead ejaculate quality to decline dramatically over time. Such may pose a particular concern in species with constrained sperm supplies, where adult males cannot replenish their sperm reserves. In eusocial hymenopterans (i.e. ants, bees and wasps with permanent castes), males typically reach adulthood with a limited quantity of sperm, and their testes deteriorate shortly thereafter. In adult life, both sexes mate during a single reproductive event. While males die quickly after copulation, they persist posthumously via sperm stored in their mates' spermatheca, sometimes for decades. As a result of this mating system, males should experience intense selection pressure to produce and transfer high‐quality sperm. Using the black garden ant (Lasius niger) as a model system, we investigated whether the duration of storage in the accessory testes before copulation affected sperm quality. We found that there was no impact of male age on sperm number or viability and that sperm DNA fragmentation decreased rather that increased with age. These results highlight the extent to which male ants, and possibly other taxa with constrained sperm supplies, have evolved extremely specialized strategies to ensure reproductive performance.

Sperm competition increases sperm production and quality in Cataglyphis desert ants.

Sperm competition is a pervasive evolutionary force that shapes sperm traits to maximize fertilization success. Indeed, it has been shown to increase sperm production in both vertebrates and invertebrates. However, sperm production is energetically costly, which may result in trade-offs among sperm traits. In eusocial hymenopterans, such as ants, mating dynamics impose unique selective pressures on ejaculate. Males are sperm limited: they enter adulthood with a fixed amount of sperm that will not be renewed. We explored whether sperm competition intensity was associated with sperm quantity and quality (i.e. sperm viability and DNA fragmentation) in nine Cataglyphis desert ants. Our results provide phylogenetically robust evidence that sperm competition is positively correlated with sperm production and sperm viability. However, it was unrelated to sperm DNA integrity, indicating the absence of a trade-off involving this trait. These findings underscore that sperm competition may strongly mould sperm traits and drive reproductive performance in eusocial Hymenoptera.

Long-term sperm storage in eusocial Hymenoptera.

In internally fertilizing species, sperm transfer is not always immediately followed by egg fertilization, and female sperm storage (FSS) may occur. FSS is a phenomenon in which females store sperm in a specialized organ for periods lasting from a few hours to several years, depending on the species. Eusocial hymenopterans (ants, social bees, and social wasps) hold the record for FSS duration. In these species, mating takes place during a single nuptial flight that occurs early in adult life for both sexes; they never mate again. Males die quickly after copulation but survive posthumously as sperm stored in their mates' spermathecae. Reproductive females, also known as queens, have a much longer life expectancy, up to 20 years in some species. Here, we review what is currently known about the molecular adaptations underlying the remarkable FSS capacities in eusocial hymenopterans. Because sperm quality is crucial to the reproductive success of both sexes, we also discuss the mechanisms involved in sperm storage and preservation in the male seminal vesicles prior to ejaculation. Finally, we propose future research directions that should broaden our understanding of this unique biological phenomenon.

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.

Indirect interactions between a native and a supposedly non-native wasp species (Hymenoptera: Vespidae: Eumeninae: Anterhynchium)

Non-native species pose a threat to native organisms. When non-native and native species are closely related, the former can often competitively exclude the latter. Many studies have focused on competitive exclusion of native insect species by non-native eusocial hymenopterans, including ants, hornets, paper wasps and bees. Although solitary species of wasps have been introduced in many regions, few studies have investigated the effects of these insects on their native congeners. We investigated competitive interactions between native and non-native solitary wasps belonging to the same genus (Hymenoptera: Vespidae: Eumeninae: Anterhynchium). Specifically, we compared resource use and natural enemies of the native Anterhynchium flavomarginatum and supposedly non-native A. gibbifrons at a forest edge in Takasago, Hyogo, Japan, in June-October 2019, using trap nests (bamboo canes). Of 950 bamboo canes, 70 (7.4%) and 50 (5.3%) were used as nests by A. flavomarginatum and A. gibbifrons, respectively. Anterhynchium flavomarginatum produced two generations over the period studied, whereas A. gibbifrons produced only one. Although A. gibbifrons began nesting two weeks after A. flavomarginatum, the nesting period of A. gibbifrons overlapped that of the first nesting period of A. flavomarginatum. Nest architecture and the inner diameter of the canes used by both species were similar, suggesting potential competition for nesting resources. Anterhynchium flavomarginatum used larvae of 14 species of moths (Lepidoptera: Crambidae, Pyralidae, Tortricidae) as food for their larval offspring, whereas A. gibbifrons used only a single species, Demobotys pervulgalis (Lepidoptera: Crambidae). Prey species were exclusive to each wasp species, indicating no competition for this resource. Three parasitoid species, Macrosiagon nasutum (Coleoptera: Ripiphoridae), Amobia distorta (Diptera: Sarcophagidae) and Megaselia sp. (Diptera: Phoridae), attacked both Anterhynchium species. The percentage parasitism by Amobia distorta was higher for the native A. flavomarginatum. Anterhynchium gibbifrons may indirectly affect A. flavomarginatum via shared parasitoids.

The Innate Immune Response of Eusocial Hymenopterans to Viral Pathogen Challenge

Abstract In recent years, insect immunology has expanded rapidly in research interest, and available literature has expanded in kind. Insects combat pathogens through a range of behavioral and physiological immune defenses. The need for robust immunity is especially important to eusocial insects; nestmate proximity increases exposure to and transmission of pathogens. Further, eusociality involves cohabitation of thousands of individuals with characteristically reduced genetic variability, in turn increasing susceptibility to epidemic disease outbreaks. To combat this, they have developed diverse responses to pathogens, including individual innate immune defenses, social immunity, and secretion of potent glandular chemicals. The range of immune responses is as diverse as the pathogens presenting the threat, and of these, viruses may present the greatest challenge. Social immunity employed by Hymenoptera and has been reviewed whereas a review has not been developed to our knowledge addressing innate immunity of eusocial Hymenopterans to viral pathogenic invaders. We argue that such a review is important to advancement of understanding of Hymenopteran biology and is critical to applied interests. We argue further that the implications of eusocial Hymenopteran innate immunity are far-reaching; their success is a source of both substantial economic loss in the case of invasive ants and significant economic gain in the case of the honey bee Apis mellifera.

A Short Review of the Venoms and Toxins of Spider Wasps (Hymenoptera: Pompilidae).

Parasitoid wasps represent the plurality of venomous animals, but have received extremely little research in proportion to this taxonomic diversity. The lion’s share of investigation into insect venoms has focused on eusocial hymenopterans, but even this small sampling shows great promise for the development of new active substances. The family Pompilidae is known as the spider wasps because of their reproductive habits which include hunting for spiders, delivering a paralyzing sting, and entombing them in burrows with one of the wasp’s eggs to serve as food for the developing larva. The largest members of this family, especially the tarantula hawks of the genus Pepsis, have attained notoriety for their large size, dramatic coloration, long-term paralysis of their prey, and incredibly painful defensive stings. In this paper we review the existing research regarding the composition and function of pompilid venoms, discuss parallels from other venom literatures, identify possible avenues for the adaptation of pompilid toxins towards human purposes, and future directions of inquiry for the field.

Caste-specific phenotypic plasticity of Asian weaver ants: Revealing the allometric and non-allometric component of female caste system of Oecophylla smaragdina (Hymenoptera: Formicidae) by using geometric morphometrics

In eusocial insects, particularly in ants, caste differentiation is extremely complicated when we rely on traditional taxonomy. In most species, the worker caste does not display any distinct morphological characters neither the caste’s central division according to their morphological size variations. We used a landmark-based geometric morphometric approach to quantify the morphological characteristics of female caste systems (queen, major and minor worker ant) of Oecophylla smaragdina. Our findings suggested that each caste has its unique shape and size. Especially in the worker caste, apart from the size variations, we can use the shape as a prominent tool for distinguishing between them. The O. smaragdina exhibits a triphasic allometry pattern. Studying the allometry and non-allometry components of each caste system revealed a highly complex size and shape relationship in the female caste systems. From the allometric and non-allometric analysis, we concluded that the major worker ants showed a closer relationship with the queen than the minor worker ant. This outcome demonstrated that Asian weaver ant exhibits complex shape variations related to size and is correlated to their functional modular characters. This research sheds new light on caste systems’ taxonomic uncertainties for eusocial hymenopteran groups, especially ants.

Social Evolution With Decoupling of Multiple Roles of Biogenic Amines Into Different Phenotypes in Hymenoptera

Convergent evolution of eusociality with the division of reproduction and its plastic transition in Hymenoptera has long attracted the attention of researchers. To explain the evolutionary scenario of the reproductive division of labor, several hypotheses had been proposed. Among these, we focus on the most basic concepts, i.e., the ovarian ground plan hypothesis (OGPH) and the split-function hypothesis (SFH). The OGPH assumes the physiological decoupling of ovarian cycles and behavior into reproductive and non-reproductive individuals, whereas the SFH assumes that the ancestral reproductive function of juvenile hormone (JH) became split into a dual function. Here, we review recent progress in the understanding of the neurohormonal regulation of reproduction and social behavior in eusocial hymenopterans, with an emphasis on biogenic amines. Biogenic amines are key substances involved in the switching of reproductive physiology and modulation of social behaviors. Dopamine has a pivotal role in the formation of reproductive skew irrespective of the social system, whereas octopamine and serotonin contribute largely to non-reproductive social behaviors. These decoupling roles of biogenic amines are seen in the life cycle of a single female in a solitary species, supporting OGPH. JH promotes reproduction with dopamine function in primitively eusocial species, whereas it regulates non-reproductive social behaviors with octopamine function in advanced eusocial species. The signal transduction networks between JH and the biogenic amines have been rewired in advanced eusocial species, which could regulate reproduction in response to various social stimuli independently of JH action.

A Review of Termite Pheromones: Multifaceted, Context-Dependent, and Rational Chemical Communications

Termite colonies, composed of large numbers of siblings, develop an important caste-based division of labor; individuals in these societies interact via intra- or intercaste chemical communications. For more than 50 years, termites have been known to use a variety of pheromones to perform tasks necessary for maintenance of their societies, similar to eusocial hymenopterans. Although trail-following pheromones have been chemically identified in various termites, other types of pheromones have not been elucidated chemically or functionally. In the past decade, however, chemical compositions and biological functions have been successfully identified for several types of termite pheromones; accordingly, the details of the underlying pheromone communications have been gradually revealed. In this review, we summarize both the functions of all termite pheromones identified so far and the chemical interactions among termites and other organisms. Subsequently, we argue how termites developed their sophisticated pheromone communication. We hypothesize that termites have diverted defensive and antimicrobial substances to pheromones associated in caste recognition and caste-specific roles. Furthermore, termites have repeatedly used a pre-existing pheromone or have added supplementary compounds to it in accordance with the social context, leading to multifunctionalization of pre-existing pheromones and emergence of new pheromones. These two mechanisms may enable termites to transmit various context-dependent information with a small number of chemicals, thus resulting in formation of coordinated, complex, and rational chemical communication systems.

Macrocyclic Lactones Act as a Queen Pheromone in a Primitively Eusocial Sweat Bee.

Eusociality is characterized by the reproductive division of labor between two castes: fertile queens and largely sterile workers. Queen pheromones are known to influence worker behavior and reproductive physiology and are therefore key components in regulating complex eusocial behavior [1]. Recent studies indicate that cuticular hydrocarbons (CHCs) act as queen pheromones in various eusocial hymenopteran species [2-8]. However, almost all species investigated to date are highly eusocial and do not include extant transitory stages from solitary to eusocial behavior [9]. Indeed, primitively eusocial species, which largely lack morphologically distinct castes, are thought to control worker reproduction through the physical aggression of the queen rather than via pheromones [10-12]. Halictid or sweat bees exhibit a high variability of eusociality including solitary and facultatively eusocial species [9, 13-16]. However, the mechanisms controlling worker reproduction in these transitory species are unknown. The results of a recent correlative study based on caste-specific chemical profiles in various halictid bees of different social levels have revealed an overproduction of macrocyclic lactones in queens compared with workers [17]. Using chemical analyses and behavioral experiments in which we simulated below-ground nests of the primitively eusocial sweat bee Lasioglossum malachurum, we identified a queen pheromone and found that macrocyclic lactones, not CHCs, influence worker behavior and decrease ovarian activation in this species. Our data suggest that the evolution of queen pheromones is more complex than previously inferred from highly eusocial species and shed new light on the complexity of the evolution of queen pheromones.

Brood parasitism in eusocial insects (Hymenoptera): role of host geographical range size and phylogeny.

Interspecific brood parasitism is common in many animal systems. Brood parasites enter the nests of other species and divert host resources for producing their own offspring, which can lead to strong antagonistic parasite-host coevolution. Here, we look at commonalities among social insect species that are victims of brood parasites, and use phylogenetic data and information on geographical range size to predict which species are most probably to fall victims to brood parasites in the future. In our analyses, we focus on three eusocial hymenopteran groups and their brood parasites: (i) bumblebees, (ii) Myrmica ants, and (iii) vespine and polistine wasps. In these groups, some, but not all, species are parasitized by obligate workerless inquilines that only produce reproductive-caste descendants. We find phylogenetic signals for geographical range size and the presence of parasites in bumblebees, but not in ants and wasps. Phylogenetic logistic regressions indicate that the probability of being attacked by one or more brood parasite species increases with the size of the geographical range in bumblebees, but the effect is statistically only marginally significant in ants. However, non-phylogenetic logistic regressions suggest that bumblebee species with the largest geographical range sizes may have a lower likelihood of harbouring social parasites than do hosts with medium-sized ranges. Our results provide new insights into the ecology and evolution of host-social parasite systems, and indicate that host phylogeny and geographical range size can be used to predict threats posed by social parasites, as well to design efficient conservation measures for both hosts and their parasites. This article is part of the theme issue 'The coevolutionary biology of brood parasitism: from mechanism to pattern'.

Spatio-Temporal Variation in Viability of Spermatozoa of Honey Bee, Apis mellifera, Drones in Central Texas Apiaries

Males of some eusocial hymenopterans live in sheltered hives where they are raised by sister workers until they are ready to mate. Large amounts of colony resources are invested in the care and nurturing of males, as they provide no contributions to colony maintenance apart from reproduction. Colonies of the honey bee, Apis mellifera L., have one queen, thousands of female workers, and a few thousand seasonal males (drones) that are reared only during the reproductive season when colony resources are plentiful. We examined the viability of spermatozoa in sexually mature drones from eight apiaries in three counties in Central Texas during the summers of 2013 and 2014. We sampled 1,622 drones from two counties in 2013, and 556 drones from three counties in 2014. Using dual fluorescent flow cytometry, viability of drone spermatozoa was measured as the proportion of total spermatozoa that was viable. The average spermatozoa viability was 46.2% in 2013 and 67.0% in 2014. We found significant variation in spermatozoa viability across apiaries, with viability in Apiary 5 significantly lower and viability in Apiary 11 significantly greater than viability in the other apiaries sampled in 2013. Likewise, males in apiaries 6 and 7 had significantly lower average viability of spermatozoa compared with other apiaries analyzed in 2014. However, the changes were not consistent across apiaries during the 2 years sampled. Our results suggested that other factors such as exposure to insecticides or seasonal availability of forage might be more important than inter-colonial genetic differences in fertility of honey bee drones during the reproductive season.

Antibacterial activity of male and female sperm-storage organs in ants.

Bacteria can damage sperm and thus reduce the reproductive success of both males and females; selection should therefore favour the evolution of antimicrobial protection. Eusocial hymenopterans might be particularly affected by such bacterial infections because of their mating ecology. In both sexes, mating is restricted to a short window early in the adult stage; there are no further chances to mate later in life. Males die shortly after mating, but queens use the acquired sperm to fertilise their eggs for years, sometimes decades. The reproductive success of both sexes is, thus, ultimately sperm-limited, which maintains strong selection for high sperm viability before and after storage. We tested the antibacterial activity of the contents of the male and female sperm-storage organs - the accessory testes and the spermatheca, respectively. As our study species, we used the bacterium Escherichia coli and the garden ant Lasius niger, whose queens can live for several decades. Our results provide the first empirical evidence that male and female sperm-storage organs display different antibacterial activity. While the contents of the accessory testes actually enhanced bacterial growth, the contents of the spermatheca strongly inhibited it. Furthermore, mating appears to activate the general immune system in queens. However, antimicrobial activity in both the spermatheca and the control tissue (head-thorax homogenate) declined rapidly post-mating, consistent with a trade-off between immunity and reproduction. Overall, this study suggests that ejaculates undergo an immune 'flush' at the time of mating, allowing storage of sperm cells free of bacteria.