Differential time allocation of foraging workers in the subterranean termite

Background

Worker reproduction has an important influence on the social cohesion and efficiency of social insect colonies, but its role in the success of invasive ants has been neglected. We used observations of 233 captive colonies, laboratory experiments, and genetic analyses to investigate the conditions for worker reproduction in the invasive Anoplolepis gracilipes (yellow crazy ant) and its potential cost on interspecific defence. We determined the prevalence of worker production of males and whether it is triggered by queen absence; whether physogastric workers with enlarged abdomens are more likely to be reproductive, how normal workers and physogastric workers compare in their contributions to foraging and defence; and whether worker-produced males and males that could have been queen- or worker-produced differ in their size and heterozygosity.

Results

Sixty-six of our 233 captive colonies produced males, and in 25 of these, some males could only have been produced by workers. Colonies with more workers were more likely to produce males, especially for queenless colonies. The average number of days between the first appearance of eggs and adult males in our colonies was 54.1 ± 10.2 (mean ± SD, n = 20). In our laboratory experiment, queen removal triggered an increase in the proportion of physogastric workers. Physogastric workers were more likely to have yolky oocytes (37–54.9%) than normal workers (2–25.6%), which is an indicator of fertile or trophic egg production. Physogastric workers were less aggressive during interspecific aggression tests and foraged less than normal workers. The head width and wing length of worker-produced males were on average 4.0 and 4.3% greater respectively than those of males of undetermined source. Our microsatellite DNA analyses indicate that 5.5% of worker-produced males and 14.3% of males of undetermined source were heterozygous, which suggests the presence of diploid males and/or genetic mosaics in A. gracilipes.

Conclusions

Our experimental work provides crucial information on worker reproduction in A. gracilipes and its potential cost to colony defence. The ability of A. gracilipes workers to produce males in the absence of queens may also contribute to its success as an invasive species if intranidal mating can take place between virgin queens and worker-produced males.

     

Response of fire ants (Formicidae: Solenopsis invicta and S.gerninata) to artificial nectars with amino acids

Abstract.

• 1 The role that amino acids in extrafloral nectars play in attracting ants to plants was investigated. Workers from laboratory colonies of Solenopsis invicta Buren and S.geminafa (F). (Formicidae) fed from artificial nectaries containing mimics of the extrafloral nectar of Passiflora menispermifoh and P.caerulea; P.menispermifoh nectar contains higher levels of amino acids (1347.3 pdml) than does the nectar of P.currulea (125.2 μm /ml).
• 2 When sugar-only and sugar—amino acid nectar mimics were presented simultaneously, more S.invicta workers were counted at sugar—amino acid nectar mimics than at sugar-only nectars. S.geminatu did not discriminate between the two nectars.
• 3 When the two Pamiflora L. nectar mimics were presented simultaneously, S.invicta and S.geminata workers were more abundant at the nectaries containing high levels of amino acids (P.menispermifolia HBK mimic) than at the nectaries containing low levels of amino acids (P.cuerulea L. mimic).
• 4 The behaviour shown by S.invicta and S.gerninata suggests that plants with high levels of amino acids in their extrafloral nectars attract more ant protectors and might suffer less herbivory than plants producing nectars with low levels of amino acids. If so, ants may favour, over evolutionary time, plants that produce nectars with high levels of amino acids.
• 5 Day-to-day variability in ant behaviour was considerable even among laboratory colonies maintained on the same diet in similar environmental conditions. This variability will reduce the selective impact that ants have on plants and may help to explain why most ant-plant interactions are facultative.

     

Effects of fertilisation on amino acid mobilisation by a plant‐manipulating insect

1. Phytophagous insects frequently manipulate their host‐plant to improve their immediate environment. This generally implies substantial modifications of host metabolism, and sometimes an alteration of nitrogen allocation within the host‐plant. However, the outcome of plant manipulation on amino acid or protein content can be modulated by environmental factors and host‐plant traits. 2. It was investigated whether the pseudogall induced by the aphid Phloeomyzus passerinii (Signoret) (Aphididae: Phloeomyzinae) in the bark of its host‐plant affects the amino acid content in bark tissues, and whether the strength of the modification is modulated by the fertilisation regime and/or the resistance level of the host‐plant. The development of aphid colonies on a resistant and a susceptible poplar genotype, under three fertilisation regimes, was studied. After the development of colonies, the free and protein‐bound amino acid content of the infested bark were quantified. 3. Fertilisation enhanced poplar growth and increased the free amino acid content of bark tissues. Infestation also triggered accumulations of both free and protein‐bound amino acids in the feeding sites, but in the susceptible genotype only. The increase in amino acid content was more pronounced when fertilisation was low, and fertilisation did not enhance aphid development. 4. In conclusion, infestation by P. passerinii triggers an accumulation of amino acids, but the effect is influenced by both the fertilisation regime and the resistance level of the host‐plant. This suggests that P. passerinii could affect the allocation of nutrients within trees during outbreaks.     

Worker reproduction and related behavior in orphan colonies of a Japanese paper wasp,Polistes jadwigae (Hymenoptera,Vespidae)

Worker reproduction and related behavior in 2 orphan colonies ofPolistes jadwigae, one of which had been maintained by a queen and 5 artificially introduced alien workers, were observed. After the queen loss, a dominance hierarchy was established among workers, and several workers laid eggs without physical interference from other workers. Only males emerged from worker-laid eggs, however, a few new queens were produced from queen-laid eggs. Investment sex ratio of queenright (=normal) colonies (0.27) fell between the 2 theoretical values, corresponding to worker control (0.20 or 0.18) and queen control (0.48 or 0.46) of the sex ratio, both being calculated by considering the excess of males produced in the orphan colonies.     

Mating structure and male production in Vespa analis and Vespa simillima (Hymenoptera: Vespidae)

We estimated queen mating frequency, genetic relatedness between workers and worker reproduction in the hornets Vespa analis and Vespa simillima using microsatellite DNA genotyping. The 20 V. analis colonies studied each contained a queen inseminated by a single male. Of the 15 V. simillima colonies studied, nine had a queen inseminated by a single male, four had a queen inseminated by two males, and two had a queen inseminated by three males. The estimated effective number of matings was 1.33 ± 0.74 (mean ± SD), with 75–85% of the offspring of the six multiply mated queens sired by single males. The values for genetic relatedness between the workers of V. analis and V. simillima were 0.739 ± 0.004 and 0.698 ± 0.013 (mean ± SD), respectively. We conclude that V. analis and V. simillima colonies are genetically monogynous and monandrous. When high relatedness between the workers occurs within colonies, kin selection theory predicts a potential conflict between queens and workers over male production. To determine whether males were derived from queens or workers, males from V. analis and V. simillima colonies were genotyped at four microsatellite loci and the level of ovary activation in workers was determined. None of the 787 V. analis workers and only 15 of 3520 V. simillima workers had developed ovaries. Furthermore, the genotyping identified no worker‐produced males in any colony. The presence of reproductive workers correlated positively with the number of workers within the colony. These results suggest that eusocial colonies with an annual life cycle tend to break down socially when they become large and are close to dying.     

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 influence of hybridization on colony structure in the ant species Temnothorax nylanderi and T. crassispinus

The parapatric sibling ant species Temnothorax nylanderi and T. crassispinus hybridize in the contact zone in the Franconian Jura, Southern Germany. Aim of our study was to investigate the impact of hybridization on colony composition and fitness. We classified colonies as either ‘pure’ or containing hybrids by determining their allozyme pattern at GPI, an enzyme that is fixed for different alleles in the two parental species, and quantified their reproductive output. Most colonies with hybrid workers had a T. crassispinus queen. Colonies with heterozygous, hybrid workers produced more young workers than colonies of the parental species but similar numbers of male and female sexuals. Female sexuals from colonies with heterozygous workers had a significantly lower weight than female sexuals from pure colonies. Only a single reproductive queen was found to be heterozygous, suggesting reduced fitness of hybrid queens. As in the parental species, hybrid colonies appear to be frequently taken over by alien queens, which obscures the genetic colony structure. Received 6 April 2006; revised 10 June 2006; accepted 15 June 2006.     

Free amino acids in the haemolymph of honey bee queens (Apis mellifera L.)

In queen honey bees the free amino acid content in the haemolymph clearly depends on the physiological function and social environment of the individual. While in drones and workers the content of free amino acids increases after emergence until it reaches a peak in 5-day-old animals and decreases afterwards, the amino acid content in queens reaches its highest level (>60 nmol/ microl haemolymph) with the onset of egg laying (10 d of age). This level is about 2.5 times more than the highest level found in workers. Queens maintain this high level also when they are older (>30 d) and continue to lay eggs in average colonies. As in drones and workers, in queens the predominant amino acid is proline, which accounts for more than 50% of the total content of free amino acids in egg-laying individuals. When 10-day-old queens are prevented from mating and do not lay eggs, their amino acid content is significantly lower compared to laying queens of the same age. Also the social environment influences the contents of free amino acids in queens. When virgin queens were kept for 6 days with 20 worker bees and sufficient honey and pollen in an incubator, they had significantly lower concentrations of amino acids than virgin queens living for the same period with about 8000 workers in a colony. Most probably, the high amino acid concentration in the haemolymph is the basis for the high protein synthesis activity of laying queens.     

The effect of workers size frequency distribution on colony development inBombus terrestris

Summary The effect of workers size frequency distribution on colony development was studied in 12 young colonies ofB. terrestris. By replacing the original workers with workers of determined size, colonies constituting small, large or mixed size nursing workers were created. The nursing workers size frequency distribution did not influence the average size of the newly emerged workers, nor their size frequency distribution. In contrast, the number of emerging workers and number of egg cells constructed by the queen in colonies with large workers were higher than in colonies with small workers. The small number of emerging workers is explained by prolonged duration of larval time in response to sub-optimal feeding in colonies of small workers. The higher number of egg cells constructed by the queens is supposed to be in response to the number of new cocoons available, or to better condition of the brood.     

The effects of colony characteristics on life span and foraging behavior of individual wasps (Polybia occidentalis,Hymenoptera: Vespidae)

Summary We studied the effects of intrinsic colony characteristics and an imposed contingency on the life span and behavior of foragers in the swarm-founding social waspPolybia occidentalis. Data were collected on marked, known-age workers introduced into four observation colonies.To test the hypothesis that colony demographic features affect worker life span, we examined the relationships of colony age and size with worker life span using survivorship analysis. Colony age and size had positive relationships with life span; marked workers from two larger, older colonies had longer life spans (¯X = 24.7 days) than those from two smaller, younger colonies (¯X = 20.1 days).We quantified the effects of experimentally imposed nest damage on forager behavior, to determine which of three predicted behavioral responses by foragers to this contingency (increased probability of foraging for building material, increased rate of foraging, or decrease in age of onset of foraging) would be employed. Increasing the colony level of need for materials used in nest construction (wood pulp and water) by damaging the nests of two colonies did not cause an increase in either the proportion of marked workers that gathered nest materials or in foraging rates of marked individuals, when compared with introduced workers in two simultaneously observed control colonies. Instead, nest damage caused a decrease in the age at which marked workers first foraged for pulp and water. The response to an increase in the need for building materials was an acceleration of behavioral development in some workers.     

Kin structure and colony male reproduction in the hornet<Emphasis Type="Italic"> Vespa crabro</Emphasis> (Hymenoptera: Vespidae)

We estimated queen mating frequency, genetic relatedness among workers, and worker reproduction in Vespa crabro flavofasciata using microsatellite DNA markers. Of 20 colonies examined, 15 contained queens inseminated by a single male, 3 colonies contained queens inseminated by two males, and 2 colonies contained queens inseminated by three males. The genetic relatedness among workers was estimated to be 0.73±0.003 (mean±SE). For this high relatedness, kin selection theory predicts a potential conflict between queens and workers over male production. To verify whether males are derived from queens or workers, 260 males from 13 colonies were genotyped at four microsatellite loci. We found that all of the males were derived from the queens. This finding was further supported by the fact that only 33 of 2,990 workers dissected had developed ovaries. These workers belonged to 2 of the 20 colonies. There was no relationship between queen mating frequency and worker reproduction, and no workers produced male offspring in any of the colonies. These results suggest that male production dominated by queens in V. crabro flavofasciata is possibly due to worker policing.     

Nestmate recognition by guards of the Asian hive bee <Emphasis Type="Italic">Apis cerana</Emphasis>

When a honey bee colony becomes queenless and broodless its only reproductive option is for some of its workers to produce sons before the colony perishes. However, for this to be possible the policing of worker-laid eggs must be curtailed and this provides the opportunity for queenless colonies to be reproductively parasitized by workers from other nests. Such reproductive parasitism is known to occur in Apis florea and A. cerana. Microsatellite analyses of worker samples have demonstrated that the proportion of non-natal workers present in an A. cerana colony declines after a colony is made queenless. This observation suggests that queenless A. cerana colonies may be more vigilant in repelling potentially parasitic non-natal workers than queenright colonies. We compared rates of nestmate and non-nestmate acceptance in both queenright and queenless A. cerana colonies using standard assays and showed that there is no statistical difference between the proportion of non-nestmate workers that are rejected in queenless and queenright colonies. We also show that, contrary to earlier reports, A. cerana guards are able to discriminate nestmate workers from non-nestmates, and that they reject significantly more non-nestmate workers than nestmate workers. Received 25 February 2008; revised 21 May 2008; accepted 25 June 2008.     

Development of a chemically defined diet for ants

A chemically defined diet is a useful tool for the study of nutritional physiology of organisms. We have developed such a diet for Camponotus carpenter ants to facilitate experiments on nutritional requirements of these ants. Worker colonies of Camponotus floridanus were fed with a chemically defined diet, containing all essential minerals, amino acids, vitamins, growth factors and sucrose in an agar matrix. After 13 weeks, neither the number of raised pupae, their dry weight, nor the mortality of workers in subcolonies fed with this diet differed significantly from control colonies fed with Bhatkar-Whitcomb-agar, in addition to cockroaches and diluted honey. Therefore, this diet is adequate for a normal brood production and a maximal growth rate of C. floridanus larvae, at least for a period of three months. This diet should be suitable for ants that are able to feed on agar-based food resources in general. Received 6 September 2006; revised 5 October 2006; accepted 18 October 2006. An erratum to this article is available at .     

Host Location Behavior in a Parasitoid of Imported Fire Ants

Female parasitoids use a hierarchy of cues to locate suitable hosts. We conducted a series of field observations and experiments to examine host location behavior in Pseudacteon tricuspis Borgmeier, a phorid parasitoid of Solenopsis invicta Buren worker ants. The parasitoids were frequently attracted to host workers at disturbed colonies, but were almost never attracted to host workers foraging at baits. When conspecific nonnestmate workers were introduced to baits, resulting in aggressive interactions, parasitoids appeared at the majority of baits. Moreover, larger numbers of parasitoids appeared at baits to which greater numbers of nonnestmate workers had been added. Addition of nonnestmate workers to disturbed colonies resulted in increased numbers of parasitoids attracted. Pseudacteon tricuspis did not display a pattern of uniform distribution at disturbed colonies but often was very abundant at some colony locations while absent or rare at nearby colony locations. Solenopsis invicta workers release alarm pheromones in aggressive interactions with nonnestmates, and this substance is likely an important chemical cue that attracts P. tricuspis flies to host workers from a distance.     

Aggression,cooperation, and relatedness among colonies of the invasive ant, <Emphasis Type="Italic">Monomorium pharaonis</Emphasis>, originating from different areas of the world

The cooperation and aggression between five laboratory colonies of Monomorium pharaonis were compared using an aggressiveness test and pupa-carrying test in laboratory arenas. The colonies were derived from field collections in different parts of Europe and USA. Generally, inter-colony aggressiveness was low and acceptance of pupae from other colonies was high. Workers from one colony (Lužiny, CZ), however, frequently displayed aggressive behavior when paired with workers from other colonies, and the Lužiny pupae were avoided by workers of other colonies in pupa-carrying tests. Behavioral tests were only partly consistent with the phylogenetic relatedness of ants because the Wisconsin colony (USA) grouped with the Lužiny colony (and not with the other three colonies) in the phylogenetic analysis but grouped with the other three colonies in the behavioral tests.     

Male production by non-natal workers in the bumblebee, Bombus deuteronymus (Hymenoptera: Apidae)

Social insect societies are considered to be composed of many extremely cooperative individuals. While workers are traditionally believed to behave altruistically, recent studies have revealed behaviors that are more selfish. One such example is intraspecific social parasitism, where workers invade conspecific colonies and produce male offspring that are reared by unrelated host workers. Such intraspecific parasitism has been reported in honeybees (Apis cerana, and A. florea) and “semi-wild” bumblebee colonies of Bombus terrestris. Here we report on intraspecific social parasitism by workers in “wild” colonies of the bumblebee B. deuteronymus. Three of the 11 B. deuteronymus colonies studied were invaded by non-natal workers, of which 75% became reproductive and produced 19% of the adult males. The invading non-natal workers produced significantly more males than resident natal workers and the non-natal brood was not discriminated against by the natal workers.     

Social parasitism by Cape honeybee workers in colonies of their own subspecies (Apis mellifera capensis Esch.)

Social parasitism is widespread in the eusocial insects. Although social parasites often show a reduced worker caste, unmated workers can also parasitize colonies. Cape honeybee workers, Apis mellifera capensis, can establish themselves as social parasites in host colonies of other honeybee subspecies. However, it is unknown whether social parasitism by laying workers also occurs among Cape honeybee colonies. In order to address this question we genotyped worker offspring of six queenless A. m. capensis colonies and determined the maternity of the reproducing workers. We found that three non-nestmate workers dominated reproduction in a host colony and produced 62.5% of the progeny. Our results show that social parasitism by laying workers is a naturally occurring part of the biology of Cape honeybees. However, such social parasitism is not frequently found (6.41% of the total worker offspring) probably due to co-evolutionary processes among A. m. capensis resulting in an equilibrium between selection for reproductive dominance in workers, colony maintenance and queen adaptation. Received 28 July 2005; revised 19 September and 11 November 2005; accepted 16 November 2005.     

The power and efficiency of brood incubation in queenless microcolonies of bumble bees (Bombus terrestris L.)

1. Ground‐nesting colonies of bumble bees incubate their brood at > 30 °C if floral forage provides sufficient energy and the thermogenic power of the colony can counteract cool soil conditions. To explore the basis of incubation, the thermogenic power and sugar consumption of orphaned nests of bumble bee workers (microcolonies) were investigated under laboratory conditions. 2. This study tested experimentally the effect of variation in worker number (ranging from four to 12 adults) on a microcolony's capacity to regulate brood temperature and recover from acute cold exposure. Microcolonies were provided with ad libitum sugar syrup and minimal insulation and maintained at an ambient temperature of c. 25 °C. Energy conversion efficiency was estimated by comparing sugar consumption with the power required for artificial incubation. The joint energetics of foraging and incubation were modelled in wild colonies to explore the effect of colony size and landscape quality on thermoregulation. 3. The results showed that all sizes of microcolonies regulated brood temperature at c. 31 °C under laboratory conditions, which required 96 mW of thermogenic power. It was estimated that individual workers of B. terrestris generated an incubatory power of 35 mW. The smallest microcolonies had the highest conversion efficiency (57%), apparently because few workers were required for incubation. 4. Modelling indicated that small microcolonies of three to seven adult workers have the capacity for normal brood incubation in the wild, but that the minimum viable colony size increases as floral forage becomes poorer or more distant. 5. These preliminary findings suggest the feasibility of identifying the minimum conditions (forage quality, soil temperature, and colony size) necessary for brood incubation by queenright colonies in the wild.     

The reproductive potential of workers in slave-making ants

Summary Colonies of slave-making ants have been used repeatedly to test sex allocation theory. It was suggested that workers of slave-making ants are more strongly selected to reproduce than workers of related, non-parasitic species, because they are incapable of manipulating sex allocation and the sexualization of larvae in their colonies. I show here that in slave-making Formicoxenini, worker ovaries on average consist of significantly more ovarioles than in non-parasiticLeptothorax species. Similarly, whereas in mostLeptothorax species, workers form reproductive hierarchies and lay eggs only in orphaned colonies, slave-maker workers show antagonistic interactions already in the presence of the queen and at least in some species have been observed ovipositing in queen-right colonies. The significance of these results is discussed.     

Nestmate Recognition Differences between Honeybee Colonies of <Emphasis Type="BoldItalic">Apis cerana</Emphasis> and <Emphasis Type="BoldItalic">Apis mellifera</Emphasis>

Nestmate recognition in Apis cerana and Apis mellifera was studied by introducing sealed queen cells heterospecifically between queenless colonies. No A. cerana queens were accepted by queenless A. mellifera; but A. mellifera queens were accepted in queenless A. cerana colonies. A. mellifera queens oviposited in queenless A. cerana colonies, but A. cerana workers removed most eggs. In time, egg removals declined, and some A. mellifera larvae that hatched from these eggs reached adulthood, and eventually about half of the workers were newly emerged A. mellifera. Eventually, the colonies consisted only of A. mellifera after A. cerana workers died by attrition. A. mellifera workers are more sensitive to nestmate recognition and killed the A. cerana virgin queens. In mixed-species colonies, after newly emerged A. mellifera workers matured, they removed eggs laid by the A. cerana queens until there were no workers to replace the old ones.