Multiple mating and facultative polygyny in the Panamanian leafcutter ant Acromyrmex echinatior

Queen mating frequency of the facultatively polygynous ant Acromyrmex echinatior was investigated by analysing genetic variation at an (AG)_n repeat microsatellite locus in workers and sexuals of 20 colonies from a single Panamanian population. Thirteen colonies were found to be monogynous, 5 colonies contained multiple queens, whereas the queen number of 2 colonies remained unresolved. Microsatellite genotypes indicated that 12 out of 13 queens were inseminated by multiple males (polyandry). The mean queen mating frequency was 2.53 and the mean genetically effective paternity frequency was 2.23. These values range among the highest found in ants, and the results are in keeping with the high mating frequencies reported for other species of leafcutter ants. Consistent skew in the proportional representation of different patrilines within colonies was found, and this remained constant in two consecutive samples of offspring. Dissections showed that all examined queens from multiple-queen colonies were mated egg-layers. The mean relatedness value among nestmate workers in polygynous colonies was lower than that for monogynous colonies. No diploid males were detected in a sample of 70 genotyped males. Worker production of males was detected in one queenless colony. We discuss our findings in relation to known patterns of multiple maternity and paternity in other eusocial Hymenoptera. Received: 2 September 1998 / Received in revised form: 3 February 1999 / Accepted: 7 February 1999     

Multiple mating in the ant Acromyrmex versicolor: a case of female control

In eusocial insects, polyandrous mating has the potential to reduce genetic relatedness of individuals within a colony, which may have a profound effect on the stability and social structure of the colony. Here we present evidence that multiple mating is common in both males and females of the desert leaf-cutter ant Acromyrmex versicolor. Females seem to have complete control over the number of matings, and mate on average with three males, even though the sperm transferred in a single copulation is sufficient to fill the spermatheca. To determine whether there is a bias in the representation of sperm from different mates in the spermatheca, females were mated to three or four males in controlled mating experiments and were subsequently allowed to found colonies in the laboratory. Paternity analysis of the offspring by random amplified polymorphic DNA analysis showed that all males that have been mated to a female successfully contributed sperm to the production of her offspring. No significant asymmetry in sperm use was detected, suggesting complete sperm mixing. Different hypotheses to explain polyandrous mating are discussed, and it is argued that the best hypothesis to explain polyandrous mating and complete sperm mixing in A. versicolor is that utilizing genetically diverse sperm confers a selective advantage on females. Received: 14 June 1995/Accepted after revision: 26 November 1995     

Seminal fluid enhances sperm viability in the leafcutter ant Atta colombica

The seminal fluid that accompanies sperm in ejaculates has been shown or suggested to affect sperm competition and paternity success of insects by preventing female remating, inducing oviposition, and forming mating plugs. In Atta leafcutter ants, queens have multiple mates but never remate later in life, although they may live and produce fertilized eggs for several decades. The mating biology and life history of these ants therefore suggests that the major function of seminal fluid is to maximize sperm viability during copulation, sperm transfer, and initial sperm storage. We tested this hypothesis by comparing the viability of testis sperm and ejaculated sperm (mixed with seminal fluid) and found a significant positive effect of seminal fluid on sperm viability. We further quantified this positive effect by adding accessory gland secretion (a major component of seminal fluid) in a dilution series, to show that minute quantities of accessory gland secretion achieve significant increases in sperm viability. Sperm stored by queens for 1 year benefited in a similar way from being exposed to accessory gland compounds after dissection in control saline solution. Our results provide the first empirical evidence that seminal fluid is important for the production of viable ejaculates and that the accessory glands of Atta males—despite their small size—are functional and produce a very potent secretion.     

Colonial recognition of fungus in the fungus-growing ant Acromyrmex subterraneus subterraneus (Hymenoptera: Formicidae)

Summary. Leaf cutting ants live in symbiosis with a basidiomycete fungus that is exploited as a source of nutrients for the ant larvae. Tests of fungus transport demonstrated that Acromyrmex subterraneus subterraneus workers discriminate concolonial fungus from alien fungus, and rejected the latter. Larvae and pupae of the ant were used as controls. Chemical analysis of the fungus revealed a great similarity between its hydrocarbon profile and that found on the ant brood. Experiments with lures showed that chemical extracts from the fungus are responsible for this discrimination process. Moreover, the presence of brood inside the fungus seemed to be important for discrimination of the fungus by workers. Resident workers accepted concolonial broodless fungus less than concolonial fungus inoculated with brood odor. Fungus seems to acquire colonial odor passively, simply by contact with the brood. The impact of fungus volume present in the nest on closure of the colony is discussed. We show here for the first time the importance of a symbiotic vegetal organism in colonial recognition in social insects. Received 14 April 2000; accepted 29 September 2000     

Waste management in the leaf-cutting ant Acromyrmex echinatior: the role of worker size, age and plasticity

Division of labour is the hallmark of the success of many social animals. It may be especially important with regard to waste management because waste often contains pathogens or hazardous toxins and worker specialisation can reduce the number of group members exposed to it. Here we examine waste management in a fungus-farming, leaf-cutting ant, Acromyrmex echinatior, in which waste management is necessary to protect their vulnerable fungal crop. By marking ants with task-specific paint colours, we found clear division of labour between workers that engage in waste management and those that forage, at least during the fine timescale of the 3-day marking period. This division of labour was influenced by both age and size, with waste management workers tending to be smaller and younger than foragers. The role of preventing contaminated ants from entering the colony was fulfilled mainly by medium-sized workers. When the level of waste was experimentally increased, most of the ants that responded to remove the waste were workers previously engaged in tasks inside the nest rather than external waste workers or foragers. These responding workers tended to be young and medium-sized. Surprisingly, the responding ants were subsequently able to revert back to working within the fungus garden, but the probability of them doing so depended on their age and the length of time they were exposed to waste. The results demonstrate the importance of division of labour with regard to waste management in A. echinatior and show that this is adaptable to changing needs.     

Serial polygyny and colony genetic structure in the monogynous queenless ant Diacamma cyaneiventre

Serial polygyny, defined as the temporal succession of several reproductive females in a colony, occurs in some monogynous social insects and has so far attracted little attention. Diacamma cyaneiventre is a queenless ponerine ant found in the south of India. Colonies are headed by one singly mated worker, the gamergate. After the death of the gamergate or her absence following colony fission, the gamergate is replaced by a newly eclosed nestmate worker. After a replacement, colonies go through short-lived periods in which two matrilines of sisters co-occur. This is a situation which can be described as serial polygyny. To measure the consequences of serial polygyny, a genetic analysis was performed on 449 workers from 46 colonies of D. cyaneiventre using five microsatellite loci. The presence of more than one matriline among workers of the same nest was detected in 19% of colonies, indicating a recent change of gamergate. The average genetic relatedness among nestmate workers was 0.751 and did not significantly differ from the theoretical expectation under strict monogyny and monandry (0.75). A simple analytical model of the temporal dynamics of serial polygyny was developed in order to interpret these results. We show that the rate of gamergate turnover relative to the rate of worker turnover is the crucial parameter determining the level of serial polygyny and its effect on the genetic structure of colonies. This parameter, estimated from our data, confirms that serial polygyny occurs in D. cyaneiventre but is not strong enough to influence significantly the average genetic relatedness among workers.     

Queen fertility, egg marking and colony size in the ant Camponotus floridanus

In ant societies, workers do not usually reproduce but gain indirect fitness benefits from raising related offspring produced by the queen. One of the preconditions of this worker self-restraint is sufficient fertility of the queen. The queen is, therefore, expected to signal her fertility. In Camponotus floridanus, workers can recognize the presence of a highly fertile queen via her eggs, which are marked with the queen's specific hydrocarbon profile. If information on fertility is encoded in the hydrocarbon profile of eggs, we expect workers to be able to differentiate between eggs from highly and weakly fertile queens. We found that workers discriminate between these eggs solely on the basis of their hydrocarbon profiles which differ both qualitatively and quantitatively. This pattern is further supported by the similarity of the egg profiles of workers and weakly fertile queens and the similar treatment of both kinds of eggs. Profiles of queen eggs correspond to the cuticular hydrocarbon profiles of the respective queens. Changes in the cuticular profiles are associated with the size of the colony the queen originates from and her current egg-laying rate. However, partial correlation analysis indicates that only colony size predicts the cuticular profile. Colony size is a buffered indicator of queen fertility as it is a consequence of queen productivity within a certain period of time, whereas daily egg-laying rate varies due to cyclical oviposition. We conclude that surface hydrocarbons of eggs and the cuticular profiles of queens both signal queen fertility, suggesting a major role of fertility signals in the regulation of reproduction in social insects.     

Construction of chimaeric gardens through fungal intercropping: a symbiont choice experiment in the leafcutter ant Atta texana (Attini, Formicidae)

Interspecies or intraspecies cooperation can be stabilized evolutionarily if choosing partners favor beneficial partners and discriminate against non-beneficial partners. We quantified such partner choice (symbiont choice) in the leafcutter ant Atta texana (Attini, Formicidae) by presenting the ants in a cafeteria-style preference assay with genotypically distinct fungal cultivars from A. texana and Acromyrmex versicolor. Symbiont choice was measured as the ants' tendency to choose one or more cultivar(s) from several pure (axenic) cultivar fragments and convert a given fungal fragment into a garden. Microsatellite DNA fingerprinting enabled us to identify the cultivars chosen by the ants for their gardens. In 91% of the choice tests, A. texana workers combined multiple cultivars into a single intercropped, chimaeric garden, and the cultivars coexisted in such chimaeric gardens for as long as 4 months. Coexistence of distinct fungal genotypes in chimaeric gardens appears to contradict a recent model of cultivar competition postulating that each cultivar secretes incompatibility compounds harming other cultivars, which presumably would preclude the intercropped polyculture observed in our experiments. Although we found no clear evidence of novel, recombinant genotypes in the experimental chimaeric gardens, the intercropping of cultivar genotypes may occasionally lead under natural conditions to exchange of genetic material between coexisting cultivars, thus introducing novel cultivar genotypes into the leafcutter symbiosis. Symbiont choice by ants and any competition between coexisting cultivar strains in chimaeric gardens do not appear to operate fast enough in our laboratory assay to convert chimaeric gardens into the monocultures observed for A. texana under natural conditions.     

Individually different foraging methods in the desert ant Cataglyphis bicolor (Hymenoptera,Formicidae)

Summary Observations and field experiments on the foraging behaviour of individual workers of Cataglyphis bicolor in a Southern Tunisian shrub desert are reported. The workers search singly for their food (mostly animal carcasses) and are singleprey loaders. The individuals differ to a great extent in their persistence to re-search the place of a find on a previous foraging excursion. The differences range continuously from thoroughly researching a place to just walking by. If, in an experiment, the same reward is offered farther from the nest, each ant persists more in re-searching the place than if food is offered close to the nest. In a further experiment, some individuals persisted less in searching near the former finding site if they had collected a fly than after collecting a piece of cheese. There is, however, evidence that individuals do not differ in their food preference. Persistent individuals, which re-search the place of a former find, are faster than non-persistent ones in retrieving food that is experimentally arranged in an aggregated manner. The experiment failed to demonstrate the (reverse) superiority of non-persistent individuals foraging on homogeneously distributed food. The observations of unmanipulated foraging excursions in the field suggest such an advantage for non-persistent foragers under natural conditions where food in general occurs widely dispersed. The colony as a whole retrieves more food within the same time from an experimental lay-out that is homogeneous than from an aggregated one. The behavioural differences between individuals could be caused by a training bias of the short-lived foragers, leading to a different assessment of the profitability of a searching method which implies returning to a formerly rewarding place. Thus, each worker uses the most promising behaviour according to its individual experience. Alternatively, the individually different searching methods could mainly contribute to the welfare of the colony as a whole rather than leading to a maximal short-term efficiency of each individual. In particular, the colony, disposing of only a few highly persistent foragers, could quickly exploit occasional short-lived, but unpredictible, clumps of food within its foraging range.     

Regional variation in queen and worker aggression in incipient colonies of the desert ant Messor pergandei

The fate of queen foundress associations in ants varies across taxa: in some, lethal fighting results in survival of a single queen, while in others, queens coexist long term. One hypothesis for this difference is that selection favors fighting when group sizes are small and tolerance when groups are large. In an experiment with the ant Messor pergandei, we formed small, medium, and large groups with newly mated queens from field populations that have different mean group sizes and differ in whether multiple queens occur in older established colonies. We found that whether queens are eliminated by fighting depends upon region of origin and not group size: regardless of co-foundress number, queens from sites with single-queen adult field colonies displayed agonistic behaviors and their colonies reduced to a single queen, while queens from sites with multiple-queen colonies did not fight and co-foundresses coexisted long term. Worker aggression towards and elimination of queens were also correlated with region of origin. Where fighting occurred, queens were as likely to be killed by workers as by other queens. An aggressive display was the most common form of agonistic interaction among queens, while fighting was relatively rare. We hypothesize that queen displays evolved in response to worker attacks because they increase the probability that workers will eliminate competitor queens. Our results suggest that the evolutionary interests of workers, as well as queens, could be important in determining the evolution and maintenance of queen elimination in foundress associations.     

Resource supplements cause a change in colony sex-ratio specialization in the mound-building ant, Formica exsecta

We examine the role of food resources on split sex ratios in Formica exsecta. Models of resource-based sex allocation predict that greater resources will cause an increase in the production of reproductive females (gynes) and an increase in overall size of offspring. We experimentally increased food resources for a subset of colonies in a polygynous population with a very male-biased sex ratio. This increase in food availability caused colonies that were male specialists the prior year to switch to female production. Overall, a significantly greater proportion of food-supplemented colonies produced gynes, compared to control colonies. Moreover, food-supplemented colonies produced significantly larger workers and males (but not gynes), compared to those produced by control colonies. There was, however, no significant difference in the numerical productivity of food-supplemented and control colonies. We also measured the natural association between colony sex specialization and proximity to conifers, which typically harbor honeydew-bearing aphids (an important natural food source). In line with the view that resources play an important role for determining sex ratios in social insects, we found that female-producing colonies were significantly closer to conifers than were male-producing colonies.     

Resource allocation,brood production and cannibalism during colony founding in the fire ant,Solenopsis invicta

Summary The colony founding characteristics of newly mated fire ant queens from monogyne colonies were studied in the field and in the laboratory under haplo- and pleometrotic conditions. Initial queen weight (live) was not correlated with subsequent progeny production. During founding, queens lost a mean of 54% of their lean weight, 73% of their fat weight and 67% of their energy content. The percentage of fat decreased from 44% to 33%. Queens lost weight or energy in relation to the amount of progeny they produced (Figs. 1, 2). The efficiency of the conversion of queen to progeny increased as more progeny were produced, leading to a decline in the unit cost of progeny (Fig. 3). The more minims a queen produced, the lower the mean weight of these minims and the faster they developed (Fig. 4). In a field experiment on pleometrotic founding, total brood increased with queen number, peaked between four and seven queens and declined with 10 queens (Fig. 5). Brood developed faster at the sunny, warmer site, but total production and queen survival was higher at the shady site. As queen density increased, production per queen decreased as a negative exponential in which the exponent estimated sensitivity of brood production to queen-crowding and the constant estimated the production by solo queens (Fig. 9). These effects of queen number were confirmed in laboratory experiments. The decrease of production per queen was small and not always detectable during the egg-laying phase, but brood attrition was always strong during the larval period and increased with queen number (Figs. 8, 10). While airborne factors may have contributed to this inhibition, most of the brood reduction was due to other causes, probably cannibalism. For a given number of minims, increased queen number increased the mean weight of these minims, an effect that resulted both from a lower minim production per queen and from cannibalism of dead queens by survivors (Fig. 11). Cannibal queens lost much less weight to produce a given number of minims than unfed control queens, and these minims were heavier (Fig. 12).     

Division of labour in a crisis: task allocation during colony emigration in the ant Leptothorax unifasciatus (Latr.)

Division of labour during colony emigration is widespread in ants. An important problem is how tasks are allocated during colony movement from one nest site to another. The generally favoured view is that emigrations are organised by a minority group of individuals, which either work unusually hard at tasks (elites) or have the exclusive task of carrying out the emigration (moving specialists). Five consecutive emigrations of a Leptothorax unifasciatus (Latr.) colony showed that the number of transporters, i.e. the individuals that took an active part in the emigration by transporting brood and ants, was smaller than it would have been if allocation of this task was random during each emigration. However, single emigrations of another three colonies, for which the spatial distribution and behaviour of the workers had been observed for a week prior to the emigration, demonstrated that the transporters did not form a homogeneous group. They differed in their spatial positions and tasks before the emigration. There was also no evidence that transporters worked harder or less hard than their nestmates before the emigration. Therefore, the individuals which carry out emigrations in L. unifasciatus colonies appear to be neither moving specialists nor elites. We propose that task allocation during emigrations of L. unifasciatus colonies is based on a feedback mechanism that involves learning.     

Experimental manipulation of colony genetic diversity had no effect on short-term task efficiency in the Argentine ant Linepithema humile

Genetic diversity might increase the performance of social groups by improving task efficiency or disease resistance, but direct experimental tests of these hypotheses are rare. We manipulated the level of genetic diversity in colonies of the Argentine ant Linepithema humile, and then recorded the short-term task efficiency of these experimental colonies. The efficiency of low and high genetic diversity colonies did not differ significantly for any of the following tasks: exploring a new territory, foraging, moving to a new nest site, or removing corpses. The tests were powerful enough to detect large effects, but may have failed to detect small differences. Indeed, observed effect sizes were generally small, except for the time to create a trail during nest emigration. In addition, genetic diversity had no statistically significant impact on the number of workers, males and females produced by the colony, but these tests had low power. Higher genetic diversity also did not result in lower variance in task efficiency and productivity. In contrast to genetic diversity, colony size was positively correlated with the efficiency at performing most tasks and with colony productivity. Altogether, these results suggest that genetic diversity does not strongly improve short-term task efficiency in L. humile, but that worker number is a key factor determining the success of this invasive species.Communicated by L. Sundström     

Chemical camouflage of the slave-making ant Polyergus samurai queen in the process of the host colony usurpation (Hymenoptera: Formicidae)

Founding queens of the obligatory social parasite ant Polyergus samurai usurp the host ant Formica japonica colony. The aggressive behaviors of F. japonica workers on the parasite queen disappear after the parasite queen kills the resident queen. To determine whether the parasite queen chemically mimics the host ants, we examined the aggressive behavior of F. japonica workers toward glass dummies applied with various extracts of the parasite queen and host workers. The crude extracts and hydrocarbon fraction reproduced the host workers’ behavior to the live ants. The extracts of the post-adoption parasite queen, as well as the nestmate extracts of F. japonica, did not elicit the aggressive behavior, but the extract of the pre-adoption parasite queen triggered attacks by the host workers. The nestmate recognition of host workers did not change, regardless of contact with the parasite. The gas chromatography and gas chromatography–mass spectrometry analyses indicated that the cuticular hydrocarbon (CHC) profile of the parasite queen drastically changed during the process of usurpation. Discriminant analysis showed the successfully usurped P. samurai queen had colony-specific CHC profiles. CHC profiles of the P. samurai queen who killed the host queen were more similar to those of the host queen than the workers, while the P. samurai queen who usurped the queenless colony had a profile similar to those of host workers. These results suggest that the P. samurai queen usually acquires the CHCs from the host queen during the fight, but from host wokers in queenless host colonies.     

Diploid male production — a significant colony mortality factor in the fire ant Solenopsis invicta (Hymenoptera: Formicidae)

Summary Two forms of the fire ant, Solenopsis invicta, occur in North America; the monogyne form has colonies with a single functional queen while the polygyne form has colonies containing many functional queens. Field surveys indicate that diploid males are common in natural populations of the polygyne form but absent from monogyne populations, in contrast to laboratory data showing that similar frequencies of queens producing such males occur in the two types of populations. Our results show that mature monogyne colonies with adopted queens rear diploid males in the laboratory, so it is unlikely that the absence of these males from monogyne colonies in the field is due to discrimination against them by monogyne workers. On the other hand, incipient monogyne colonies that produce diploid males exhibit significantly higher mortality and significantly slower rates of growth (Figs. 1–3) than colonies producing workers only. These results suggest that the observed distribution of male diploidy in S. invicta can be explained by differential mortality of diploid male producing colonies of the two forms, with such colonies of the monogyne form experiencing 100% mortality early in development. The mortality differences due to this factor are shown to be related to the different social structures and modes of colony founding characterizing the two forms.     

Food influence on colonial recognition and chemical signature between nestmates in the fungus-growing ant <Emphasis Type="Italic">Acromyrmex subterraneus subterraneus</Emphasis>

Summary. Neotropical Fungus-growing leaf-cutting ants (tribe Attini) live in obligatory symbiosis with a fungus, which they grow on fresh leaves harvested by workers. Colonial recognition is likely based on chemical cues provided by cuticular hydrocarbons that have been found to be partly influenced by environmental odor sources. The diet breadth of Acromyrmex subterraneus subterraneus enabled us to test the impact of different plant diets on colonial recognition. The intermediary of the fungus in the ants feeding habit adds a special angle to the question. From a queenright (QR) mother colony of A. s. subterraneus we formed several groups of queenless (QL) workers with fungus (approx. 700 ants). The QR colony and two of the QL-groups were fed with the same diet of fresh bramble leaves. Two other QL-groups were fed with privet leaves and two with rose flowers. After 4 months, QR workers were significantly more aggressive towards the QL-group fed with rose flowers or privet leaves than towards workers of the QL-groups fed with fresh bramble leaves. Rose-fed QL workers were aggressive towards privet-fed QL workers and vice versa, but never towards workers of their counterpart group that fed on the same diet. These results suggest that the absence of the queen or the separation time between groups played a minor role in shaping nestmate recognition cues as compared to the diet. The behavioral studies were supplemented by chemical analyses of cuticles, postpharyngeal glands (PPG) and plant-food extracts revealing profiles variations that were correlated with the dietary changes. However, although the plant extract contained several hydrocarbons there was no congruency between the plant profile and the respective diet-group ants. These results support the hypothesis that the diet influences indirectly the chemical profiles and consequently the recognition cues in A. s. subterraneus.     

Colony growth and the ontogeny of worker polymorphism in the fire ant,Solenopsis invicta

Summary Colony size and worker polymorphism (headwidth) were determined for fire ant colonies ranging from incipient to 12 years of age. Colonies grew approximately logistically, reaching half size between 21/2 and 31/2 yr and reaching their maximum size of about 220000 workers after 4 to 6 yr. Colony size showed strong seasonal variation. There was some evidence that growth rate may vary with food density. Incipient colonies are monomorphic and consist of small workers only, but as colonies grow, production of larger workers causes the size-frequency distributions to become strongly skewed. These skewed distributions were shown to consist of two slightly overlapping normal distributions, a narrow one defined as the minor workers, and a much broader one defined as the major workers. Major workers differ from minor workers in having been subjected to a discrete, additional stimulation of body growth, resulting in a second normal subpopulation. The category of media is seen to be developmentally undefined. The mean headwidth of the workers in both of these subpopulations increased during the first 6 mo. of colony life, until colonies averaged about 4000 workers. Headwidth of minors declined somewhat in colonies older than about 5 yr, but that of majors remained stable. When the first majors appear, their weight averages about twice that of minors. This increases to about 4 times at 6 mo. and remains stable thereafter. The range of weights of majors is up to 20 times that of minors. Growth of the subpopulation of major workers is also logistic, but more rapid than the colony as a whole, causing the proportion of major workers to increase with colony size. In full sized colonies, about 35% of the workers are majors. Total biomass investment in majors increases as long as colonies grow, beginning at about 10% at 2 months and reaching about 70% in mature colonies. This suggests that major workers play an important role in colony success. The total dry biomass of workers peaked at about 106 g, that of majors at about 72 g. These values then fluctuate seasonally in parallel to number of workers. When colony growth ceases, the proportion of majors remains approximately stable. Colony size explained 98% of the variation in the number of major workers.This is paper No. 18 of the Fire Ant Research Team