Host–parasite genotypic interactions in the honey bee: the dynamics of diversity

Parasites are thought to be a major driving force shaping genetic variation in their host, and are suggested to be a significant reason for the maintenance of sexual reproduction. A leading hypothesis for the occurrence of multiple mating (polyandry) in social insects is that the genetic diversity generated within‐colonies through this behavior promotes disease resistance. This benefit is likely to be particularly significant when colonies are exposed to multiple species and strains of parasites, but host–parasite genotypic interactions in social insects are little known. We investigated this using honey bees, which are naturally polyandrous and consequently produce genetically diverse colonies containing multiple genotypes (patrilines), and which are also known to host multiple strains of various parasite species. We found that host genotypes differed significantly in their resistance to different strains of the obligate fungal parasite that causes chalkbrood disease, while genotypic variation in resistance to the facultative fungal parasite that causes stonebrood disease was less pronounced. Our results show that genetic variation in disease resistance depends in part on the parasite genotype, as well as species, with the latter most likely relating to differences in parasite life history and host–parasite coevolution. Our results suggest that the selection pressure from genetically diverse parasites might be an important driving force in the evolution of polyandry, a mechanism that generates significant genetic diversity in social insects.     

Variation in patriline reproductive success during queen production in orphaned colonies of the thelytokous ant Cataglyphis cursor

In genetically diverse insect societies (polygynous or polyandrous queens), the production of new queens can set the ground for competition among lineages. This competition can be very intense when workers can reproduce using thelytoky as worker lineages that manage to produce new queens gain a huge benefit. Selection at the individual level might then lead to the evolution of cheating genotypes, i.e. genotypes that reproduce more than their fair share. We studied the variation in reproductive success among worker patrilines in the thelytokous and highly polyandrous ant Cataglyphis cursor. Workers produce new queens by thelytoky in orphaned colonies. The reproductive success of each patriline was assessed in 13 orphaned colonies using genetic analysis of 433 workers and 326 worker-produced queens. Our results show that patrilines contributed unequally to queen production in half of the colonies, and the success of patrilines was function of their frequencies in workers. However, over all colonies, we observed a significant difference in the distribution of patrilines between workers and worker-produced queens, and this difference was significant in three of 13 colonies. In addition, six colonies contained a low percentage of foreign workers (drifters), and in one colony, they produced a disproportionably high number of queens. Hence, we found some evidence for the occurrence of rare cheating genotypes. Nevertheless, cheating appears to be less pronounced than in the Cape Honey bee, a species with a similar reproductive system. We argue that worker reproduction by parthenogenesis might not be common in natural populations of C. cursor.     

Does genetic diversity hinder parasite evolution in social insect colonies?

Polyandry is often difficult to explain because benefits of the behaviour have proved elusive. In social insects, polyandry increases the genetic diversity of workers within a colony and this has been suggested to improve the resistance of the colony to disease. Here we examine the possible impact of host genetic diversity on parasite evolution by carrying out serial passages of a virulent fungal pathogen through leaf-cutting ant workers of known genotypes. Parasite virulence increased over the nine-generation span of the experiment while spore production decreased. The effect of host relatedness upon virulence appeared limited. However, parasites cycled through more genetically diverse hosts were more likely to go extinct during the experiment and parasites cycled through more genetically similar hosts had greater spore production. These results indicate that host genetic diversity may indeed hinder the ability of parasites to adapt while cycling within social insect colonies.     

Differential resistance across paternal genotypes of honey bee brood to the pathogenic bacterium Melissococcus plutonius

Melissococcus plutonius is a pathogenic bacterium affecting immature stages of the western honey bee (Apis mellifera) and leads to European foulbrood (EFB) disease. Despite EFB outbreaks increasing in frequency in several countries in recent decades, there is little knowledge on the epidemiology of M. plutonius or on the defence mechanisms of honey bees against this pathogen. Mating of honey bee queens with multiple males (polyandry) can be such a mechanism, as it has been shown to be beneficial to colony health and fitness. It is hypothesized that a high level of polyandry was selected for in response to pathogen pressure to maximize the probability that at least some patrilines among nestmates in a colony possess a high degree of resistance to specific pathogens, ultimately protecting colonies against infections. We show that M. plutonius infection provokes differential mortality among patrilines of immature honey bee workers. Such differences indicate a genetic origin of resistance against this pathogen—supporting the polyandry hypothesis—and open up avenues to improve control of EFB disease via selective breeding.     

Inbred and outbred honey bees (Apis mellifera) have similar innate immune responses

Social insect colonies provide ideal conditions for the spread of pathogens. It has been proposed that the extreme polyandry and genetic diversity seen in the colonies of some eusocial insect species is central to a colony’s defence against disease. Indeed, empirically, colonies headed by polyandrous queens have lower incidence of pathogens than genetically uniform monoandrous colonies. The mechanisms of improved resistance in genetically diverse colonies could arise from the genetic diversity among worker genotypes or from increased innate immunity arising from heterozygosity at immune gene loci within individual workers. Here, we investigate the effects of heterozygosity on two components of the honey bee (Apis mellifera) innate immune system: encapsulation and phenoloxidase (PO) activity. No significant effect of heterozygosity on immune system activity was evident for either encapsulation or PO activity. Thus, we conclude that while encapsulation and PO activity are important components of the immune response, it seems that they do not underlie the positive effects of genetic diversity on parasite and pathogen resistance in honey bees.     

Intracolonial genetic variability in honeybee larval resistance to the chalkbrood and American foulbrood parasites

The origin of multiple mating of queens in social Hymenoptera is a widely debated topic in evolutionary biology. One of the hypotheses is that genetic variability would benefit the colony by increasing its resistance to parasites through various mechanisms. One among the predictions of this hypothesis is that the resistance of different patrilines within a colony to parasites of different species should be independent, as a result of independent gene-for-gene interactions with each parasite. To test this aspect of the hypothesis, two honeybee colonies (Apis mellifera) were infected with the fungus Ascosphaera apis and two colonies with both A. apis and the American foulbrood bacterium Paenibacillus larvae. Patrilines were found to vary in resistance of larvae to A. apis in all four colonies, but similar variation in resistance was not found to P. larvae. Common resistance to both pathogens was not detected. This study supports the hypothesis that polyandry in social insects could have originated as an adaptation to decrease the impact of diseases.     

Ployandry versus polygyny versus parasites

Although social insect colonies are most easily conceptualized as consisting of a single, once-mated queen and her worker progeny, the number of queens per colony and the number of times queens mate varies broadly in ants and other social insects. Various hypotheses have been suggested for the resulting range of breeding systems and social organizations, respectively; one set of hypotheses relating to both queen number and mate number at the same time is a need for genetic variation, especially in relation to disease resistance. We here carry out a comparative analysis using phylogenetic information and, contrary to one non-phylogenetic previous study, we find that polyandry and polygyny are not significantly associated. However, the level of relatedness within colonies, a quantity affected by both polyandry and polygyny, is significantly associated with parasite loads: species with colonies with low relatedness levels have lower parasite loads. Given that, under the variance-reduction principle, selection on queens for mating frequency ought to continue even in polygynous colonies, we suggest that while parasite loads indeed seem to correlate with intra-colony genetic variability, the relationship to polyandry and polygyny may be complex and requires considerably more experimental investigation.     

Genetic variation within social insect colonies reduces parasite load

In colonies of social Hymenoptera (ants, bees and wasps), workers are often not very closely related to each other, because queens mate with several different males (polyandry) or because several functional queens are present (polygyny). Both characteristics increase genetic variation among the queens'' reproductive and worker offspring, but the benefits of this increased variation remain obscure. Here we report an experiment where genetically homogeneous and genetically heterogeneous colonies of the bumble bee, Bombus terrestris, have been exposed to parasitism under field conditions. Colonies of high or low genetic variation were achieved by adding and removing brood from donor colonies. The results showed a consistent effect in that genetically variable colonies experienced reduced parasite loads, i.e. lower prevalence, intensity and parasite species richness, for a range of protozoa, nematodes, mites or parasitoids affecting the workers. We therefore propose that polyandry and/or polygyny of social insects may be beneficial under parasitism.     

Worker genetic diversity and infection by Nosema apis in honey bee colonies.

The hypothesis that parasites and pathogens select for polyandry in eusocial Hymenoptera was tested, using the honey bee Apis mellifera and its microsporidian parasite Nosema apis. Five honey bee colonies with low and five with high worker genetic diversity were infected with N. apis spores. At 54-56 days after inoculation, parasite spores in the workers' midguts were counted to determine whether there was a greater variation of infection intensity (spore counts per worker) in high-diversity colonies than in low-diversity ones. In all colonies there were two discrete sets of workers, with few or many parasite spores. To compare the variations of infection intensity between two colony groups, coefficients of variation were calculated for all workers examined, and for the slightly infected and strongly infected workers. The percentages of slightly infected workers in the low- and high-diversity groups were also compared. None of the comparisons between low- and high-diversity colonies showed significant differences, therefore no relation was found between honey bee workers' genetic diversity and their infection with N. apis.     

The evolution of polyandry: intrinsic sire effects contribute to embryo viability

Females typically mate with more than one male despite the costs incurred, thus questioning Bateman's principle. A series of genetic benefits have been proposed to account for the evolution of polyandry, including the acquisition of viability genes for offspring. The 'intrinsic male quality' hypothesis suggests that polyandry increases the probability that females produce offspring sired by males that bestow high viability on their offspring. Heritable variation in viability is the basic requirement for the occurrence of this genetic benefit. By using a half-sib breeding design with a species of cricket in which polyandry is known to increase hatching success, we present clear experimental evidence that intrinsic male quality contributes to embryo viability. Despite recent support for the evolution of polyandry based on compatibility of genotypes between males and females, we show that hatching success is not determined by an interaction between paternal and maternal genotypes but rather that sons inherit paternal genes that influence the viability of eggs laid by their mates. Moreover, our data implicate a potential role for indirect genetic effects of male accessory gland products on embryo viability. Additive genetic contributions to embryo viability may be an important factor underlying the frequently observed benefits of polyandrous behaviour.     

No selection for change in polyandry under experimental evolution

What drives mating system variation is a major question in evolutionary biology. Female multiple mating (polyandry) has diverse evolutionary consequences, and there are many potential benefits and costs of polyandry. However, our understanding of its evolution is biased towards studies enforcing monandry in polyandrous species. What drives and maintains variation in polyandry between individuals, genotypes, populations and species remains poorly understood. Genetic variation in polyandry may be actively maintained by selection, or arise by chance if polyandry is selectively neutral. In Drosophila pseudoobscura, there is genetic variation in polyandry between and within populations. We used isofemale lines to found replicate populations with high or low initial levels of polyandry and tracked polyandry under experimental evolution over seven generations. Polyandry remained relatively stable, reflecting the starting frequencies of the experimental populations. There were no clear fitness differences between high versus low polyandry genotypes, and there was no signature of balancing selection. We confirmed these patterns in direct comparisons between evolved and ancestral females and found no consequences of polyandry for female fecundity. The absence of differential selection even when initiating populations with major differences in polyandry casts some doubt on the importance of polyandry for female fitness.     

The social consequences of honey bee polyandry: the effects of kinship on worker interactions within colonies

Honey bees, Apis mellifera L., are polyandrous and several males simultaneously father offspring within a single colony. The relatedness of female colony members therefore varies with their paternity: workers encounter both patrilineal full sister (r¯=0·75) and non-patrilineal half-sister (r¯=0·25) nestmates. The impact of this intra-colony genetic variation on social grooming and trophallaxis (liquid food exchange) among workers in colonies consisting of two phenotypically-distinct worker patrilines was examined. Workers in these colonies groomed and fed a disproportionately large number of full sisters despite a tendency to encounter a disproportionately large number of half-sisters. Thus, workers actively discriminated between full and half-sisters. This patrilineal discrimination occurred both in colonies with laying queens and in a queenless colony rearing replacement queens. These results suggest that intracolony genetic variation may have a major effect on colony social organization.     

Good genes and the maternal effects of polyandry on offspring reproductive success in the bulb mite

Genetic benefits are potentially the most robust explanation of the controversial issue of evolutionary maintenance of polyandry, but the unambiguous demonstration of such benefits has been hindered by the possibility of their confusion with maternal effects. Previous research has shown that polyandrous bulb mite females produce daughters with higher fecundity than monandrous females. Here, we investigate whether this effect arises because polyandrous females invest more in their offspring, or because their offspring inherit 'good genes' from their fathers. Females were mated with either one or four (different) males. However, by sterilizing three of the four males with ionizing radiation, we eliminated any chance of sexual selection (in the polyandrous treatment) so that any differences in the female mating regimes must have been owing to maternal effects. Polyandry had no significant effect on daughter fecundity, thus indicating that any previously documented effects must have been genetic. This was further supported by a significant association between fathers' offensive sperm-competitive ability and the fecundity of their daughters. The association with fathers' sperm defensive ability was not significant, and neither was the association between fathers' sperm competitiveness and sons' reproductive success. However, sons of polyandrous females had lower reproductive success than sons of monandrous females. This shows that the maternal effects of polyandry should be taken into account whenever its costs and benefits are being considered.     

Material and genetic benefits of female multiple mating and polyandry

The maintenance of female polyandry has traditionally been attributed to the material (direct) benefits derived from male mating resources (e.g. nuptial gifts) accrued by multiple mating. However, genetic (indirect) benefits offer a more robust explanation since only polyandrous, not monandrous, females may gain both material benefits from multiple mating and genetic benefits from multiple sires. Discriminating between material and genetic benefits is essential when addressing the mechanism by which polyandry is adaptively maintained, but are difficult to disentangle because they affect fitness in similar ways. To test the hypothesis that genetic benefits maintain polyandry, we compared four components of fitness (longevity, fecundity, hatching success and survivorship) between monandrous and polyandrous females in the ground cricket, Allonemobius socius. We discovered that females derived nongenetic benefits from mating multiply, in that the magnitude of the nuptial gift was positively associated with the number of eggs produced. However, polyandrous females had over a two-fold greater hatching success and a 43% greater offspring survivorship, leading to a significantly higher relative fitness than the monandrous strategy. These results were independent of the confounding effects of material benefits, implying that genetic contributions play a large role in the maintenance of polyandry and potentially in the antagonistic coevolutionary relationship between polyandry and male nuptial gifts. Copyright 2002 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour     

Reproductive conflicts in polyandrous and polygynous ant Formica sanguinea

The occurrence of multiple reproductives within an ant colony changes the balance between indirect fitness benefits and reproductive competition. We test whether the number of matings by an ant queen (polyandry) correlates negatively with the number of reproductive queens in the colony (polygyny), whether the patrilines and matrilines differ in their contribution to the sexual and worker progeny and whether there is an overall reproductive skew. For these aims, we genotyped both worker and sexual offspring from colonies of the ant Formica sanguinea in three populations. Most colonies were monogynous, but eight (11%) were polygynous with closely related queens. Most queens in the monogynous colonies (86%) had mated with multiple males. The effective paternity was lower than the actual number of mates, and the paternity skew was significant. Furthermore, in some monogynous colonies, the patrilines were differently represented in the worker pupae and sexual pupae produced at the same time. Likewise, the matrilines in polygynous colonies were differently present in worker pupae and male offspring. The effective number of matings by a queen was significantly lower in polygynous colonies (mean m_e = 1.68) than in monogynous colonies (means 2.06–2.61). The results give support to the hypotheses that polyandry and polygyny are alternative breeding strategies and that reproductive competition can lead to different representation of patrilines and matrilines among the sexual and worker broods.     

Condition dependence of reproductive strategy and the benefits of polyandry in a viviparous lizard

Species in which males do not contribute to reproduction beyond the provision of sperm offer good opportunities to study the potential genetic benefits that females can obtain from polyandry. Here, we report the results of a study examining the relationships between polyandry and components of female fitness in the common lizard (Lacerta vivipara). We found that polyandrous females produce larger clutches than monandrous females. Polyandrous females also lose fewer offspring during the later stages of gestation and at birth, but we did not find any relationship between polyandry and physical characteristics of viable neonates. Our results were consistent with the predictions of the intrinsic male quality hypothesis, while inbreeding avoidance and genetic incompatibility avoidance might also explain some part of the variation observed in clutch size. Moreover, the benefits of polyandry appeared to depend on female characteristics, as revealed by an interaction between reproductive strategy and female length on reproductive success. Thus, all females did not benefit equally from mating with multiple males, which could explain why polyandry and monandry coexist.     

Queen promiscuity lowers disease within honeybee colonies

Most species of social insects have singly mated queens, but in some species each queen mates with numerous males to create a colony with a genetically diverse worker force. The adaptive significance of polyandry by social insect queens remains an evolutionary puzzle. Using the honeybee (Apis mellifera), we tested the hypothesis that polyandry improves a colony's resistance to disease. We established colonies headed by queens that had been artificially inseminated by either one or 10 drones. Later, we inoculated these colonies with spores of Paenibacillus larvae, the bacterium that causes a highly virulent disease of honeybee larvae (American foulbrood). We found that, on average, colonies headed by multiple-drone inseminated queens had markedly lower disease intensity and higher colony strength at the end of the summer relative to colonies headed by single-drone inseminated queens. These findings support the hypothesis that polyandry by social insect queens is an adaptation to counter disease within their colonies.     

Genetic variation changes the interactions between the parasitic plant-ecosystem engineer Rhinanthus and its hosts

Within-species genetic variation is a potent factor influencing between-species interactions and community-level structure. Species of the hemi-parasitic plant genus Rhinanthus act as ecosystem engineers, significantly altering above- and below-ground community structure in grasslands. Here, we show the importance of genotypic variation within a single host species (barley-Hordeum vulgare), and population-level variation among two species of parasite (Rhinanthus minor and Rhinanthus angustifolius) on the outcome of parasite infection for both partners. We measured host fitness (number of seeds) and calculated parasite virulence as the difference in seed set between infected and uninfected hosts (the inverse of host tolerance). Virulence was determined by genetic variation within the host species and among the parasite species, but R. angustifolius was consistently more virulent than R. minor. The most tolerant host had the lowest inherent fitness and did not gain a fitness advantage over other infected hosts. We measured parasite size as a proxy for transmission ability (ability to infect further hosts) and host resistance. Parasite size depended on the specific combination of host genotype, parasite species and parasite population, and no species was consistently larger. We demonstrate that the outcome of infection by Rhinanthus depends not only on the host species, but also on the underlying genetics of both host and parasite. Thus, genetic variations within host and parasite are probably essential components of the ecosystem-altering effects of Rhinanthus.     

Modelling the role of intracolonial genetic diversity on regulation of brood temperature in honey bee (Apis mellifera L.) colonies

In polyandrous social insects such as honey bees, a worker’s affinity for a particular task may be genetically infl uenced and so some patrilines may have lower stimulus thresholds for commencing a task than others. We used simulation models to investigate the effects of intracolonial diversity in the task thresholds that stimulate workers to engage in heating and cooling during nest thermoregulation. First, we simulated colonies comprised of one or 15 patrilines that were engaged in heating the brood nest, and observed that single patriline colonies maintained, on average, less stable brood nest temperatures than multiple patriline colonies. Second we simulated colonies with five patrilines that were engaged in cooling their nest, recording the proportions of bees of different patrilines that engaged in nest cooling in response to changing temperatures. Both of our simulations show remarkably similar qualitative patterns to those that we have previously observed empirically. This provides further support for the hypothesis that geneticallybased variability in task thresholds among patrilines within honey bee colonies is an important contributor to the ability of colonies to precisely thermoregulate their nests, and we suggest that diversity is important for optimal expression of a range of other colony-level phenotypes. Received 17 June 2005; revised 27 October 2005; accepted 23 December 2005.     

Caste-specific expression of genetic variation in the size of antibiotic-producing glands of leaf-cutting ants

Social insect castes represent some of the most spectacular examples of phenotypic plasticity, with each caste being associated with different environmental conditions during their life. Here we examine the level of genetic variation in different castes of two polyandrous species of Acromyrmex leaf-cutting ant for the antibiotic-producing metapleural gland, which has a major role in defence against parasites. Gland size increases allometrically. The small workers that play the main role in disease defence have relatively large glands compared with larger workers, while the glands of gynes are substantially larger than those of any workers, for their body size. The gland size of large workers varies significantly between patrilines in both Acromyrmex echinatior and Acromyrmex octospinosus. We also examined small workers and gynes in A. echinatior, again finding genetic variation in gland size in these castes. There were significant positive relationships between the gland sizes of patrilines in the different castes, indicating that the genetic mechanism underpinning the patriline variation has remained similar across phenotypes. The level of expressed genetic variation decreased from small workers to large workers to gynes. This is consistent with the hypothesis that there is individual selection on disease defence in founding queens and colony-level selection on disease defence in the worker castes.