Workers' sons rescue genetic diversity at the sex locus in an invasive honey bee population

The hallmark of eusociality is the division of labour between reproductive (queen) and nonreproductive (worker) females. Yet in many eusocial insects, workers retain the ability to produce haploid male offspring from unfertilized eggs. The reproductive potential of workers has well‐documented consequences for the structure and function of insect colonies, but its implications at the population level are less often considered. We show that worker reproduction in honey bees can have an important role in maintaining genetic diversity at the sex locus in invasive populations. The honey bee sex locus is homozygous‐lethal, and, all else being equal, a higher allele number in the population lead to higher mean brood survival. In an invasive population of the honey bee Apis cerana in Australia, workers contribute significantly to male production: 38% of male‐producing colonies are queenless, and these contribute one‐third of all males at mating congregations. Using a model, we show that such male production by queenless workers will increase the number of sex alleles retained in nascent invasive populations following founder events, relative to a scenario in which only queens reproduce. We conclude that by rescuing sex locus diversity that would otherwise be lost, workers' sons help honey bee populations to minimize the negative effects of inbreeding after founder events and so contribute to their success as invaders.     

Mating frequency, genetic structure, and sex ratio in the intermorphic female producing ant species Myrmecina nipponica

1. Myrmecina nipponica has two types of colonies: a queen colony type, in which the reproductive females are queens and new colonies are made by independent founding, and an intermorphic female colony type, in which reproductive females belong to a wingless intermediate morphology between queen and worker, and where colonies multiply through colonial budding. 2. The mating frequencies of reproductive females in both types indicate monoandry. The relatedness among nestmates in both types was almost 0.75, however relatedness between mother and daughter in intermorphic female colonies was slightly higher than that of queen colonies. 3. The sex ratio (corrected investment female ratio) was 0.70 at the population level, suggesting that the sex ratio is controlled by workers in this species, however the ratio differed greatly between the two types of colonies. Queen colonies (n = 37) had a female‐biased sex ratio of 0.77 while intermorphic female colonies (n = 33) had a ratio of 0.56. 4. Each reproductive intermorphic female was accompanied by an average of 2.9 workers (including virgin intermorphic females) in the colonial budding, and when the investment to those workers was added to the female investment, the sex ratio reached 0.81. 5. The frequency distribution of sex ratio was bimodal, with many colonies producing exclusively males or females, however mean estimated relatedness within colonies was almost 0.75. These data are inconsistent with the genetic variation hypothesis, which is one of the predominant hypotheses accounting for the between‐colony variation in sex ratio.     

The inhibiting effect of the queen bee (Apis mellifera L.) foot-print pheromone on the construction of swarming queen cups

Experiments were conducted to determine the rôle of population density of queenright honey bee colonies, and that of the queen bee pheromonal secretions, on the induction and inhibition of swarming queen cup construction during swarming and non-swarming seasons. Construction of queen cups was induced experimentally in overcrowded queenright colonies, during winter, which is a non-swarming season. This construction was induced by high population density of bee workers: above a threshold of 2.3 bee workers/ml there was a relationship between the number of cups constructed and the colony density. During the swarming season a relationship was established between the free volume of a hive (population density) and the number of queen cups constructed: 1.5 cups in a colony that occupied 80,960 ml, compared with 77 cups in a colony hived within a volume of 20, 240 ml. Observations of the queen's movements upon combs in colonies of high and normal population densities showed that in an overcrowded colony the queen bee was almost absent from the bottom edges of the comb, where queen swarming cups and cells are constructed. The tarsal glands of queens are located in the fifth tarsomere and the glandular oily secretion is deposited by the foot-pads upon the combs surface. The rate of secretion by the queen's tarsal glands was about 13 times higher than by those of the workers. A bioassay for testing the inhibitory effects of the queen's glandular extracts on the construction of queen cups was developed. It was based on increasing worker bee population densities, and can be used effectively throughout the year in a subtropical climate.The application of tarsal and mandibular glands' secretion to comb bottom edges in overcrowded colonies (bioassay) caused the inhibition of queen cup construction. None of these two secretions affected construction of these cups when applied separately. We presume that due to colony overcrowding the queen bee is unable to deposit the non-volatile secretions from tarsal glands along the comb edges and that the deficiency of the foot-print pheromone triggers the construction of swarming cups along the non-inhibited areas.     

A quantitative study of worker reproduction in queenright colonies of the Cape honey bee, Apis mellifera capensis

Reproduction by workers is rare in honey bee colonies that have an active queen. By not producing their own offspring and preventing other workers from producing theirs, workers are thought to increase their inclusive fitness due to their higher average relatedness towards queen-produced male offspring compared with worker-produced male offspring. But there is one exception. Workers of the Cape honey bee, Apis mellifera capensis, are able to produce diploid female offspring via thelytokous parthenogenesis and thus produce clones of themselves. As a result, worker reproduction and tolerance towards worker-produced offspring is expected to be more permissive than in arrhenotokous (sub)species where worker offspring are male. Here we quantify the extent to which A. m. capensis workers contribute to reproduction in queenright colonies using microsatellite analyses of pre-emergent brood. We show that workers produced 10.5% of workers and 0.48% of drones. Most of the workers' contribution towards the production of new workers coincided with the colonies producing new queens during reproductive swarming.     

Intraspecific queen parasitism in a highly eusocial bee

Insect societies are well-known for their advanced cooperation, but their colonies are also vulnerable to reproductive parasitism. Here, we present a novel example of an intraspecific social parasitism in a highly eusocial bee, the stingless bee Melipona scutellaris. In particular, we provide genetic evidence which shows that, upon loss of the mother queen, many colonies are invaded by unrelated queens that fly in from unrelated hives nearby. The reasons for the occurrence of this surprising form of social parasitism may be linked to the fact that unlike honeybees, Melipona bees produce new queens in great excess of colony needs, and that this exerts much greater selection on queens to seek alternative reproductive options, such as by taking over other nests. Overall, our results are the first to demonstrate that queens in highly eusocial bees can found colonies not only via supersedure or swarming, but also by infiltrating and taking over other unrelated nests.     

Physiological consequences of prolonged nursing in the honey bee

Honey bee workers normally produce brood food at an age of 5 to 15 days. However, natural events like swarming or brood diseases may lead to the occurrence of over-aged nurses. Here we investigated the physiological consequences of prolonged nursing for both the nurses and the brood they rear, and tried to separate the effects of chronological age and of task affiliation on some important physiological parameters. Brood was reared in groups of colonies with either a normal age structure or with moderately over-aged workers.The haemolymph concentrations of total protein and vitellogenin, the development of mandibular and hypopharyngeal glands, and the activity of α-glucosidase in the hypopharyngeal glands of nurses from these groups of colonies were compared. Moreover, we used the fertility of young workers reared by normal- and overaged nurses as a bioindicator for the quality of the brood care they had received. It showed that parameters linked to the production of brood food proteins remained stable in over-aged nurses, whereas the development of mandibular glands regressed. Workers reared by over-aged nurses had more ovarioles and showed stronger ovary development under queenless conditions. Our results indicate that while over-aged nurses remain capable of producing brood food, they are not functionally equivalent to young nurses. The partial degeneration of the mandibular glands normally occurring at the end of the nursing period cannot be prevented by prolonged nursing. The distinct phenotype of workers reared by old nurses raises the question of possible age-related specialisations among nurses in colonies with a normal age structure. Received 14 July 2008; revised 18 November 2008; accepted 21 November 2008.     

Prediction of social structure and genetic relatedness in colonies of the facultative polygynous stingless bee Melipona bicolor (Hymenoptera, Apidae)

Stingless bee colonies typically consist of one single-mated mother queen and her worker offspring. The stingless bee Melipona bicolor (Hymenoptera: Apidae) shows facultative polygyny, which makes this species particularly suitable for testing theoretical expectations concerning social behavior. In this study, we investigated the social structure and genetic relatedness among workers from eight natural and six manipulated colonies of M. bicolor over a period of one year. The populations of M. bicolor contained monogynous and polygynous colonies. The estimated genetic relatedness among workers from monogynous and polygynous colonies was 0.75 ± 0.12 and 0.53 ± 0.16 (mean ± SEM), respectively. Although the parental genotypes had significant effects on genetic relatedness in monogynous and polygynous colonies, polygyny markedly decreased the relatedness among nestmate workers. Our findings also demonstrate that polygyny in M. bicolor may arise from the adoption of related or unrelated queens.     

Inbreeding and reproductive investment in the ant Formica exsecta

In social animals, inbreeding depression may manifest by compromising care or resources individuals receive from inbred group members. We studied the effects of worker inbreeding on colony productivity and investment in the ant Formica exsecta. The production of biomass decreased with increasing inbreeding, as did biomass produced per worker. Inbred colonies produced fewer gynes (unmated reproductive females), whereas the numbers of males remained unchanged. As a result, sex ratios showed increased male bias, and the fraction of workers increased among the diploid brood. Males raised in inbred colonies were smaller, whereas the weight of gynes remained unchanged. The results probably reflect a trade-off between number and quality of offspring, which is expected if the reproductive success of gynes is more dependent on their weight or condition than it is for males. As males are haploid (with the exception of abnormal diploid males produced in very low frequencies in this population), and therefore cannot be inbred themselves, the effect on their size must be mediated through the workers of the colony. We suggest the effects are caused by the inbred workers being less proficient in feeding the growing larvae. This represents a new kind of social inbreeding depression that may affect sex ratios as well as caste fate in social insects.     

圆唇散白蚁补充生殖蚁的类型与建群能力

摘要: 【目的】为了探讨圆唇散白蚁Reticulitermes labralis补充生殖蚁对巢群稳定和发展的作用。【方法】对野外巢群进行调查研究, 及对婚飞成虫通过雌雄配对与补充生殖蚁隔离巢群进行人工饲养的对比研究。【结果】在野外巢群只发现1对原始蚁王蚁后, 而补充生殖蚁的数量最多可达到689头/巢。圆唇散白蚁有3种类型的补充生殖蚁, 即由工蚁转化来的无翅型补充生殖蚁、若蚁转化来的翅芽型补充生殖蚁和末龄若蚁羽化来的拟成虫型补充生殖蚁。实验室条件下婚飞配对群体和隔离群体建群1个月后的存活率分别为64%和96%。建巢初期婚飞配对群体的子代数目增长缓慢, 2个月时的群体数量为6.3±1.54, 10个月时的群体数量也仅为8.4±1.47; 而隔离建群补充生殖蚁2个月时的群体数量为52.4±6.44, 10个月时的群体数量为164.3±20.85, 都高于婚飞配对群体。 此外, 野外巢群的补充蚁后跟原始蚁后一样都具有发达的卵巢。【结论】在圆唇散白蚁中补充生殖蚁是白蚁巢群主要的繁殖力量, 也是建立新巢群的重要繁殖品级。     

Worker reproductive parasitism in naturally orphaned colonies of the Asian red dwarf honey bee, Apis florea

The truce between honey bee (Apis spp.) workers over reproduction is broken in the absence of their queen. Queenright workers generally abstain from personal reproduction, raising only the queen’s offspring. Queenless workers activate their ovaries, produce eggs, and reduce the rate at which they destroy worker-laid eggs, so that some eggs are reared to maturity. Reduced policing of worker-laid eggs renders queenless nests vulnerable to worker reproductive parasitism (WRP), and may result in the colony raising eggs of unrelated (non-natal) workers that parasitize it. Queenless colonies of A. florea are heavily parasitized with the eggs of non-natal workers. However, queenless colonies often abscond upon disturbance and build a small comb in which to rear their own male offspring. We investigated three naturally occurring orphaned colonies to determine if they are also parasitized. We show that WRP is present in orphaned colonies, and non-natal workers have significantly higher rates of ovary activation than natal workers. In contrast to experimentally manipulated colonies, in our samples, natal and non-natal workers had statistically equal reproductive success, but this may have been due to the small number of non-natals present.     

Genetic analysis of spatial foraging patterns and resource sharing in bumble bee pollinators

Conservation biologists, evolutionary ecologists and agricultural biologists require an improved understanding of how pollinators utilize space and share resources. Using microsatellite markers, we conducted a genetic analysis of space use and resource sharing at several spatial scales among workers of two ecologically dissimilar bumble bee species (Bombus terrestris and B. pascuorum) foraging in an urban landscape (London, UK). At fine scales, the relatedness of workers visiting small patches of flowers did not differ significantly from zero. Therefore, colonies shared flower patches randomly with other colonies, suggesting that worker scent-marks deterring visits to unrewarding flowers have not evolved as signals benefiting nestmates. To investigate space use at intermediate scales, we developed a program based on Thomas & Hill's maximum likelihood sibship reconstruction method to estimate the number of colonies utilizing single sites. The average number of colonies (95% confidence limits) sending workers to forage at sites of approximately 1 ha in area was 96 colonies (84-118) in B. terrestris and 66 colonies (61-76) in B. pascuorum. These values are surprisingly high and suggested that workers travelled far from their colonies to visit the sites. At the landscape scale, there was little or no genetic differentiation between sites. We conclude that urban habitats support large bumble bee populations and are potentially valuable in terms of bumble bee conservation. In addition, bumble bee-mediated gene flow in plants is likely to occur over large distances and plant-bumble bee conservation requires landscape-scale action.     

Cheaters sometimes prosper: targeted worker reproduction in honeybee (Apis mellifera) colonies during swarming

Kin selection theory predicts that honeybee (Apis mellifera) workers should largely refrain from producing their own offspring, as the workers collectively have higher inclusive fitness if they rear the sons of their mother, the queen. Studies that have quantified levels of ovary activation and reproduction among workers have largely supported this prediction. We sampled pre‐emergent male pupae and adult workers from seven colonies at regular intervals throughout the reproductive part of the season. We show that the overall contribution of workers to male (drone) production is 4.2%, nearly 40 times higher than is generally reported, and is highest during reproductive swarming, when an average of 6.2% of the males genotyped are worker‐produced. Similarly, workers in our samples were 100 times more likely to have active ovaries than previously assumed. Worker reproduction is seasonally influenced and peaks when colonies are rearing new queens. Not all worker subfamilies contribute equally to reproduction. Instead, certain subfamilies are massively over‐represented in drone brood. By laying eggs within the period in which many colonies produce virgin queens, these rare worker subfamilies increase their direct fitness via their well‐timed sons.     

Reproductive strategy of the slave antFormica podzolica relative to raiding efficiency of enslaver species

Summary Formica podzolica serves as host to slave-making ants in North America. We propose thatF. podzolica may respond to slavery by two alternative colony-growth and reproductive strategies depending on the raiding ability of the slavemaker: (1) Rapid colony growth at the expense of producing sexuals to a stage where raiding by unspecialized, facultative slavemakers, capable of exploiting only small colonies, becomes unlikely owing to a strong work force and (2) Early production of sexual offspring at the cost of colony growth to secure some sexual production in an environment with specialized obligate enslavers, capable of raiding large colonies. We tested the strategies by excavating 30 small to moderately large mounds ofF. podzolica and measured reproductive parameters of colonies in relation to mound size, worker number, and worker size. Mound area predicted worker number satisfactorily. Worker number correlated significantly with worker head width and with number of worker and sexual offspring. With a growing work force, the proportion of sexual offspring increased in the total offspring. Two thirds of the colonies producing sexuals emitted single sex, sex being independent of colony size. Some of the large colonies produced both sexes with a strong bias toward either sex. The unweighted population-level sex ratio did not differ from even, being 0.52 (numerical) or 0.54 (biomass). Very large mounds (not excavated) had small workers and highly male-biased sex ratios, probably owing to energy constraints set by central-place foraging. Population-level colony ontogeny data did not fit either one of the suggested strategies, but imply a mixture of the two. We discuss an alternative, still untested raid-independent explanation to the ontogeny pattern.     

The role of queen mandibular pheromone and colony congestion in honey bee (Apis mellifera L.) reproductive swarming (Hymenoptera: Apidae)

The roles of honey bee queen mandibular pheromone and colony congestion in the inhibition of swarming were investigated. Two colony siz.es were used: small, congested colonies and large, uncongested colonies. Both groups of colonies were treated with various dosages of the five-component, synthetic queen mandibular pheromone in the spring, and the extent and timing of swarming were followed. Most treatment groups received pheromone or a solvent blank (control) on a stationary slide; one group of the congested colonies received a pheromone treatment via an aerosol spray. The pheromone was not effective at delaying swarming in the congested colonies at any dosage applied on slides, but the aerosol spray-treated colonies swarmed significantly later in the season than the control colonies. The uncongested, pheromone-treated colonies exhibited a dose-dependent delay in swarming, with the highest dosage colonies swarming almost four weeks later than the control colonies. These results indicate an interaction between congestion and pheromone in the control of honey bee reproduction. While congestion may in itself be a factor stimulating swarming, these results are consistent with the interpretation that colony congestion reduces the transmission of queen pheromone within the nest, thereby removing the queen 's pheromone-based inhibition of queen rearing and subsequent swarming by workers.     

Queen-controlled sex ratios and worker reproduction in the bumble bee Bombus hypnorum,as revealed by microsatellites

Social insect colonies provide model systems for the examination of conflicts among parties with different genetic interests. As such, they have provided the best tests of inclusive fitness theory. However, much remains unknown about in which party's favour such conflicts are resolved, partly as a result of the only recent advent of the molecular tools needed to examine the outcome of these conflicts. Two key conflicts in social insect colonies are over control of the reproductive sex ratio and the production of male offspring. Most studies have examined only one of these conflicts but in reality they occur in tandem and may influence each other. Using microsatellite analyses, the outcome of conflict over sex ratios and male production was examined in the bumble bee, Bombus hypnorum. The genotypes were determined for mother queens, their mates and males for each of 10 colonies. In contrast to other reports of mating frequency in this species, all of the queens were singly mated. The population sex ratio was consistent with queen control, suggesting that queens are winning this conflict. In contrast, workers produced over 20% of all males in queen-right colonies, suggesting that they are more effective in competing over male-production. Combining these results with previous work, it is suggested that worker reproduction is a labile trait that may well impose only small costs on queen fitness.     

Social life-history response to individual immune challenge of workers of Bombus terrestris L.: a possible new cooperative phenomenon

Parasites typically reduce host survival or fecundity. To minimize fitness loss, hosts can make temporal adjustments of their reproductive effort. To date such plastic shifts of life‐history traits in response to parasitism are only known from solitary organisms where infected individuals can react by themselves. In the case of social insects, where brood care and reproductive effort is shared between reproductive individuals (typically the queen) and workers, adjustments of the reproductive effort would depend on collective decision‐making. We tested for this possibility by experimentally activating the immune response of individual workers in colonies of the bumblebee, Bombus terrestris L. This induction resulted, in combination with environmental conditions, in a reduction of fitness of the social unity (i.e. colony success, measured by number and biomass of offspring) and a collective response towards earlier reproduction. As both phenomena are expressed at the level of the colony, the result suggests that key elements of the use of immune defence have been maintained through the evolutionary transition to sociality.     

Hemolymph juvenile hormone titers in worker honey bees under normal and preswarming conditions

Swarming is an important mechanism by which honey bee, Apis mellifera L., colonies reproduce, yet very little is known about the physiological changes in workers that are preparing to swarm. In this study, we determined the endocrine status of worker honey bees in preswarming colonies and in normal (nonswarming) colonies. Juvenile hormone (JH) titers in worker bees were similar in both groups before queen cells were present, but they became significantly lower in preswarming colonies compared with normal colonies when queen cells occurred in preswarming colonies. The lower JH titers in the preswarming colonies suggest that behavioral development is delayed in these colonies, consistent with previous reports that preswarming colonies have reduced foraging activities. Understanding the endocrine status of bees preparing for swarming will help us to better understand the biology of swarming.     

COLONY SEX RATIOS,CONFLICT BETWEEN QUEENS AND WORKERS,AND APPARENT QUEEN CONTROL IN THE ANT PHEIDOLE DESERTORUM

Sex-ratio conflict between queens and workers was explored in a study of colony sex ratios, relatedness, and population investment in the ant Pheidole desertorum. Colony reproductive broods consist of only females, only males, or have a sex ratio that is extremely male biased. Colonies producing females (female specialists) and colonies producing males (male specialists) occur at near equal frequency in the population. Most colonies apparently specialize in producing one reproductive sex throughout their life. Allozyme analyses show that relatedness does not differ within male-specialist and female-specialist colonies and they do not appear to differ in available resources. In the population, workers are nearly three times more closely related to females than males; however, the investment sex ratio is near equal (1.01, female/male), which is consistent with queen control. Selection should be strong on workers to increase investment in reproductive females, so why do workers in male-specialist colonies produce only (or nearly only) males? One hypothesis is that queens in male-specialist colonies prevent the occurrence of reproductive females, perhaps by producing worker-biased female eggs. An earlier simulation study of genetic evolution of sex ratios in social Hymenoptera (Pamilo 1982b) predicts that such mechanisms can result in the evolution of bimodal colony sex ratios and queen control. Results on P. desertorum are generally consistent with that study; however, information is not currently available to test some of the model's predictions and assumptions.     

Size and composition of swarming colonies in Provespa anomala (Hymenoptera, Vespidae), a nocturnal social wasp

Summary: Ninety-eight nests of various growing stages of Provespa anomala were found in the lowlands of Sumatra, Indonesia, from August to January. Its colony cycle is considered to be annual, with nests built throughout the year. There were two types of early nests before the emergence of workers, one with a worker number of less than 63 and the other of more than 231. The former were new nests, each containing a new queen and were built by reproductive swarming, whereas each of the latter was possibly re-built by an old queen and an absconding swarm. Ten swarming colonies were attracted by an oil-lamp light; these consisted of a copulated queen with immature oocytes in the ovary and 55 workers on average. No males, or workers stylopized by Xenos provesparum were observed either in colonies before worker emergence or in swarms coming flying into the light. Differences between swarming in Provespa, polygyny and nest relocation in Vespa, and those in swarm-founding Polistinae are discussed.     

Queen replacement in African and European honey bee colonies with and without afterswarms

We examined the dynamics of the queen replacement process in African and European colonies that did and did not produce afterswarms. In colonies without afterswarms, the queen replacement process was completed in 24–48 hours, the first-emerging virgin queen (VQ) typically inherited the natal nest even if multiple queens emerged, workers performed few vibration signals on emerged queens, and all signaling activity was directed toward early emerging VQs. In contrast, if colonies did produce afterswarms, the queen replacement process required 5–6 days, there was no advantage for first-emerging queens, vibration rates on emerged queens were 25 times greater, and signaling activity was directed toward all VQs. Although vibration signal activity was more pronounced in colonies with afterswarms, the signal was consistently associated with increased VQ survival under all conditions. These trends were exhibited similarly in the African and European colonies, suggesting that they have broad applicability to queen-replacement decisions over a range of environmental and racial conditions. However, the African and European colonies differed in the total number of queens involved in the elimination process and the relative importance of queen duels and pre-emergence destruction under the different reproductive strategies. Taken together, our results suggest that worker behavior is a major determinant for the outcome of queen replacement, either through reduced interactions that allow first-emerged queens to rapidly eliminate rivals, or through increased use of interactions such as the vibration signal, which may allow workers to influence the ultimate fate of each emerged VQ. We discuss the possibility that these behavior patterns may reflect the roles of cooperation and conflict in shaping honey bee reproductive decisions. Received 8 May 2007; revised 7 November 2007; accepted 20 November 2007.