Chemical tools to distinguish the fire ant species Solenopsis invicta and S. saevissima (Formicidae: Myrmicinae) in Southeast Brazil

Morphologically similar fire ants Solenopsis invicta and Solenopsis saevissima are broadly sympatric in southeastern Brazil. Chemistry from venom (2,6-dialkyl piperidine alkaloids) and cuticular hydrocarbons have been reported as potentially important tools for differentiating Solenopsis species. We have analysed two chemical classes in widely separated populations of S. invicta and S. saevissima and find that both piperidine alkaloids and cuticular hydrocarbons separate the two species. Piperidine alkaloids clustered S. invicta but not S. saevissima. Cuticular hydrocarbons strongly clustered both S. invicta and S. saevissima. One population morphologically identified as S. invicta presented piperidine alkaloids and cuticular hydrocarbons markedly different from either species. The distinctive piperidine alkaloid differences among populations of S. saevissima and the marked difference in piperidine alkaloid and hydrocarbon profiles of the anomalous population of S. invicta suggest undescribed species fire ant in southeastern Brazil.     

Traits allowing some ant species to nest syntopically with the fire ant Solenopsis saevissima in its native range

Supercolonies of the red fire ant Solenopsis saevissima (Smith) develop in disturbed environments and likely alter the ant community in the native range of the species. For example, in French Guiana only 8 ant species were repeatedly noted as nesting in close vicinity to its mounds. Here, we verified if a shared set of biological, ecological, and behavioral traits might explain how these 8 species are able to nest in the presence of S. saevissima. We did not find this to be the case. We did find, however, that all of them are able to live in disturbed habitats. It is likely that over the course of evolution each of these species acquired the capacity to live syntopically with S. saevissima through its own set of traits, where colony size (4 species develop large colonies), cuticular compounds which do not trigger aggressiveness (6 species) and submissive behaviors (4 species) complement each other.     

Impact of native vegetation cover near crops on the occurrence and molecular diversity of fire ants

Solenopsis invicta and Solenopsis saevissima are the most common fire ants in Brazil. Environmental disturbances favour the colony dispersal of both species, particularly those with an anthropogenic origin. However, the frequencies of the species can vary according to the amount of anthropogenic disturbances. Crops near native vegetation fragments is a common landscape in the Brazilian Atlantic Forest. Here, we analyse if there is influence of native vegetation cover on the occurrence of these fire ants and their molecular diversity in crops. Our hypothesis is that these species are antagonistic in terms of occurrence and molecular diversity when compared in the same habitat. We selected crops near fragments with around 50% of native vegetation cover, a percentage informative enough to detect biological responses from insects belonging to a same order. Nests were collected and the species were identified using external morphology, and mtDNA COI sequences. We reject our hypothesis, which suggest that the species colonize the vegetation patches around crops in a similar way. Solenopsis invicta is not limited by vegetation cover, and has a high haplotype diversity when compared to S. saevissima, particularly when the vegetation cover is greater than 50%. Additionally, the vegetation cover seems to not have influence in the total number of haplotypes, but the species have haplotypes that are exclusive to each landscape. Our findings suggest that S. invicta is expanding to native vegetation areas that it has not occupied before, as S. saevissima was dominant there.     

A South American fire ant,Solenopsis nr. saevissima,in Guadeloupe,French West Indies

Several species of fire ants, well known for their painful stings, have spread from their native ranges and have become invasive pests, notably Solenopsis geminata, Solenopsis invicta, and Solenopsis richteri. Here, I report the first known exotic spread of another fire ant, Solenopsis nr. saevissima, a South American species now established on the French West Indian island of Guadeloupe. In 2008, I first found this shiny yellow fire ant at Plage de Viard beach on Guadeloupe. Genetic analyses indicated that this species was close to the yellow–brown variant of S. saevissima known only from Western Amazonia, but not an exact match. I returned to Guadeloupe in 2011 and found S. nr. saevissima at Plage de Viard plus five additional sites along a >20 km stretch on the east coast of Basse-Terre, from the Jarry commercial shipping port in the north to Plage Roseau in the south. It seems most likely that S. nr. saevissima arrived in Guadeloupe from South America in a commercial shipment unloaded at the port in Jarry. It is unclear whether this species will develop into a major new problem. Solenopsis nr. saevissima may simply replace another pernicious fire ant, S. geminata, which apparently arrived in Guadeloupe from South America several 100 years ago.     

Reactions by army ant workers to nestmates having had contact with sympatric ant species

It was recently shown that Pheidole megacephala colonies (an invasive species originating from Africa) counterattack when raided by the army ant, Eciton burchellii. The subsequent contact permits Pheidole cuticular compounds (that constitute the “colony odour”) to be transferred onto the raiding Eciton, which are then not recognised by their colony-mates and killed. Using a simple method for transferring cuticular compounds, we tested if this phenomenon occurs for Neotropical ants. Eciton workers rubbed with ants from four sympatric species were released among their colony-mates. Individuals rubbed with Solenopsis saevissima or Camponotus blandus workers were attacked, but not those rubbed with Atta sexdens, Pheidole fallax or with colony-mates (control lot). So, the chemicals of certain sympatric ant species, but not others, trigger intra-colonial aggressiveness in Eciton. We conclude that prey-ant chemicals might have played a role in the evolution of army ant predatory behaviour, likely influencing prey specialization in certain cases.     

Genetic clusters and sex-biased gene flow in a unicolonial Formica ant

Background  

Animal societies are diverse, ranging from small family-based groups to extraordinarily large social networks in which many unrelated individuals interact. At the extreme of this continuum, some ant species form unicolonial populations in which workers and queens can move among multiple interconnected nests without eliciting aggression. Although unicoloniality has been mostly studied in invasive ants, it also occurs in some native non-invasive species. Unicoloniality is commonly associated with very high queen number, which may result in levels of relatedness among nestmates being so low as to raise the question of the maintenance of altruism by kin selection in such systems. However, the actual relatedness among cooperating individuals critically depends on effective dispersal and the ensuing pattern of genetic structuring. In order to better understand the evolution of unicoloniality in native non-invasive ants, we investigated the fine-scale population genetic structure and gene flow in three unicolonial populations of the wood ant F. paralugubris.     

Global-scale analyses of chemical ecology and population genetics in the invasive Argentine ant

Ants are some of the most abundant and ecologically successful terrestrial organisms, and invasive ants rank among the most damaging invasive species. The Argentine ant is a particularly well-studied invader, in part because of the extreme social structure of introduced populations, known as unicoloniality. Unicolonial ants form geographically vast supercolonies, within which territorial behaviour and intraspecific aggression are absent. Because the extreme social structure of introduced populations arises from the widespread acceptance of conspecifics, understanding how this colonymate recognition occurs is key to explaining their success as invaders. Here, we present analyses of Argentine ant recognition cues (cuticular hydrocarbons) and population genetic characteristics from 25 sites across four continents and the Hawaiian Islands. By examining both hydrocarbon profiles and microsatellite genotypes in the same individual ants, we show that native and introduced populations differ in several respects. Both individual workers and groups of nestmates in the introduced range possess less diverse chemical profiles than ants in the native range. As previous studies have reported, we also find that introduced populations possess much lower levels of genetic diversity than populations in the native range. Interestingly, the largest supercolonies on several continents are strikingly similar to each other, suggesting that they arose from a shared introduction pathway. This high similarity suggests that these geographically far-flung ants may still recognize and accept each other as colonymates, thus representing distant nodes of a single, widely distributed supercolony. These findings shed light on the behaviour and sociality of these unicolonial invaders, and pose new questions about the history and origins of introduced populations.     

A type of unicoloniality within the native range of the fire ant Solenopsis saevissima

To determine if a type of unicoloniality exists in the fire ant Solenopsis saevissima in its native range, we conducted intraspecific aggressiveness tests in French Guiana between workers originating from 15 human-disturbed sites. We identified two "colonial groups" spread over 54 km and 12.5 km, respectively. Workers from the same group never attacked each other, but inter-group confrontations resulted in a high level of aggressiveness. These large "colonial groups" enhances the threat occasioned by S. saevissima for both agriculture and the environment.     

Stealthy invaders: the biology of Cardiocondyla tramp ants

Many invasive ant species, such as the Argentine ant or the red imported fire ant, have huge colonies with thousands of mass-foraging workers, which quickly monopolise resources and therefore represent a considerable threat to the native ant fauna. Cardiocondyla obscurior and several other species of this myrmicine genus have similarly been transferred throughout the tropics by human activities. However, because their colonies are tiny and workers forage solitarily, Cardiocondyla are often not recognized as successful invaders. Here, we document that the life history of Cardiocondyla closely resembles that of the more conspicuous tramp species, with polygyny, intranidal mating, budding, worker sterility, low genetic variability, and possibly also unicoloniality. Given that introduced Cardiocondyla may locally reach a very high population density, the effects of these stealthy invaders on the native arthropod fauna should receive more attention. Received 18 May 2005; revised 23 September 2005; accepted 29 September 2005.     

Cuticular hydrocarbons in workers of the slave-making ant Polyergus samurai and its slave, Formica japonica (Hymenoptera: Formicidae)

Comparisons of cuticular hydrocarbons between workers of the dulotic ant Polyergus samurai and its slave, Formica japonica, were carried out. Gas chromatography–mass spectrometry showed that the slave‐maker and its slave shared the major cuticular hydrocarbon compounds, but possessed several minor products unique to each species. No difference in hydrocarbon composition was detected between enslaved and free‐living F. japonica workers, suggesting that association with P. samurai has no qualitative effect on hydrocarbon composition in these ants. Principal component analyses of the cuticular hydrocarbon profiles (CHP) revealed that (i) CHP was species specific in a given mixed colony; and (ii) among mixed colonies, P. samurai workers had species‐colony specific CHP, while the same feature was not always found in enslaved and free‐living F. japonica workers. Therefore, a ‘uniform colony odor’ in terms of CHP is not achieved in naturally mixed colonies of P. samurai nor those of its slaves, F. japonica.     

The role of cuticular hydrocarbons as chemical cues for nestmate recognition in the invasive Argentine ant (<Emphasis Type="Italic">Linepithema humile</Emphasis>)

Argentine ants (Linepithema humile) in their native South American range, like most other ant species, form spatially restricted colonies that display high levels of aggression toward other such colonies. In their introduced range, Argentine ants are unicolonial and form massive supercolonies composed of numerous nests among which territorial boundaries are absent. Here we examine the role of cuticular hydrocarbons (CHCs) in nestmate recognition of this highly damaging invasive ant using three supercolonies from its introduced range. We conducted behavioral assays to test the response of Argentine ants to workers treated with colonymate or non-colonymate CHCs. Additionally, we quantified the amount of hydrocarbons transferred to individual ants and performed gas chromatography-mass spectrometry (GC/MS) to qualitatively characterize our manipulation of CHC profiles. The GC/MS data revealed marked differences in the hydrocarbon profiles across supercolonies and indicated that our treatment effectively masked the original chemical profile of the treated ants with the profile belonging to the foreign individuals. We found that individual workers treated with foreign CHCs were aggressively rejected by their colonymates and this behavior appears to be concentration-dependent: larger quantities of foreign CHCs triggered higher levels of aggression. Moreover, this response was not simply due to an increase in the amount of CHCs applied to the cuticle since treatment with high concentrations of nestmate CHCs did not trigger aggression.The results of this study bolster the findings of previous studies on social insects that have implicated CHCs as nestmate recognition cues and provide insight into the mechanisms of nestmate recognition in the invasive Argentine ant. Received 6 February 2007; revised 31 May and 27 July 2007; accepted 16 August 2007.     

Disruption of ant-seed dispersal mutualisms by the invasive Asian needle ant (<Emphasis Type="Italic">Pachycondyla chinensis</Emphasis>)

By disrupting the structure of native ant assemblages, invasive ants can have effects across trophic levels. Most studies to date, however, have focused on the impacts just two species (Linepithema humile and Solenopsis invicta). The impacts of many other invasive ant species on ecological processes in their introduced range are unknown. In this study we tested the hypothesis that the invasive ant Pachycondyla chinensis disrupts ant-seed dispersal mutualisms by displacing native ant species, especially the keystone mutualist Aphaenogaster rudis, while failing to disperse seeds itself. In a paired design we measured the impact of P. chinensis on the native ant-plant seed dispersal mutualism. The number of A. rudis workers was 96% lower in invaded than in intact plots, and the number of seeds removed was 70% lower in these plots. Finally, in invaded plots the abundance of Hexastylis arifolia, a locally abundant myrmecochorous plant, was 50% lower than in plots where P. chinensis was absent. A parsimonious interpretation of our results is that P. chinensis causes precipitous declines in the abundance of A. rudis within invaded communities, thereby disrupting the ant-plant seed dispersal mutualisms and reducing abundances of ant-dispersed plants. In sum, the magnitude of the effects of P. chinensis on seed dispersal is quantitatively similar to that documented for the intensively studied invasive Argentine ant. We suggest that more studies on the impacts of less-studied invasive ant species on seed dispersal mutualisms may increase our knowledge of the effects of these invaders on ecosystem function.     

Potential Global Range Expansion of the Invasive Fire Ant, Solenopsis invicta

The red imported fire ant, Solenopsis invicta Buren, is an invasive pest that has become widespread in the southern United States and Caribbean after introduction from South America in the 1930s. This species, which has diverse detrimental impacts on recipient communities, was recently discovered in Australia and New Zealand and has the potential to colonize numerous other regions. We used a dynamic, ecophysiological model of colony growth to predict the potential global range expansion of this invasive species. Based on minimum and maximum daily temperatures, the model estimates colony alate production and predicts future geographic range limits. Because S. invicta populations are limited by arid conditions as well as cold temperatures, we superimposed precipitation data upon temperature-based predictions, to identify regions that do not receive enough rainfall to support this species across the landscape. Many areas around the globe, including large portions of Europe, Asia, Africa, Australia, and numerous island nations, are at risk for S. invicta infestation. Quarantine officials should be vigilant for any accidental introductions of this pest in susceptible regions. Costs of eradication increase dramatically as the area of infestation grows, and large infestations may be impossible to eradicate. Other South American Solenopsis fire ants (e.g., S. richteri Forel) may become invasive if the opportunity arises, and our predictions for S. invicta may approximate the potential range limits for these species as well.     

Fire ants: What do rural and urban areas show us about occurrence, diversity,and ancestral state reconstruction?

In South America, Solenopsis saevissima and S. invicta are the most common fire ants. Nests are founded in areas under anthropic interference like urban or rural areas, but S. invicta is found preferentially in those with the greatest anthropic interference. However, we do not know the rates at which they exist in anthropized areas next to high density of native vegetation. Areas with 60 to 90% of native Atlantic Forest were selected to verify the occurrence of both species in rural and urban areas. We investigated the molecular diversity and applied the reconstruction of the ancestral state analysis for each species. A total of 186 nests were analyzed and we found that the two species had the same proportion in the urban area. However, S. saevissima had a higher rate of prevalence in the rural area, in addition to having a greater number of haplotypes and ancestry associated with this type of habitat for the region. S. invicta had the same number of haplotypes in both rural and urban regions, and less haplotypic diversity. We conclude that S. saevissima is a species typically associated with rural areas and S. invicta, although present, is not dominant in urban areas. Keywords: Biodiversity, haplotype diversity, DNA barcoding, mtDNA     

Selective preying of the sphecid wasp Trachypus boharti on the meliponine bee Scaptotrigona postica: potential involvement of caste‐specific cuticular hydrocarbons

The specialist digger wasp Trachypus boharti Rubio‐Espina preys exclusively on males of the stingless bee Scaptotrigona postica Latreille 1807, although the hunting attacks involve both male and worker bees of S. postica and members of its own species. To understand the mechanism of prey selection, the cuticular hydrocarbon patterns of workers and males of S. postica are analyzed in detail, and the mandibular secretion of males is examined. The cuticular profiles of males and workers are distinctively different. The major group of cuticular compounds, heptacosene isomers, is twice as abundant in workers as in males. There is no clear distinction between worker and male mandibular secretions. Such a distinct and straightforward caste‐specific difference in cuticular hydrocarbons could function as a recognition cue by which T. boharti distinguishes between workers and males of S. postica.     

Experimental Evidence of Large-Scale Unicoloniality in the Tramp Ant Wasmannia auropunctata (Roger)

We examined intraspecific colonial aggressiveness in Wasmannia auropunctata (Roger), a tramp species originating from the neotropics. By observing the results of one-on-one confrontations, we compared the behavioral responses of workers originating from six New Caledonian locations (introduced range) and four Brazilian cocoa plantations (original range). We recorded interindividual aggressive behavior on four levels ranging from physical contact, with no aggressive response, to prolonged aggressiveness, including stinging by one or both ants. In Brazil, we often observed high intraspecific aggressiveness between populations originating from distant locations, indicating that W. auropunctata may behave as a multicolonial species in its native range. In New Caledonia, paired encounters resulted in low agonistic behavior, as shown by the absence of full attacks (which include stinging by one or both opponents). Our results suggest that W. auropunctata behaves as a single supercolony throughout New Caledonia and that the scale of its unicoloniality (widespread colonies with interconnected nests without aggressiveness between workers originating from distant areas) is different in introduced and native populations. According to the present study, it seems likely that differences in intraspecific aggressiveness between native and introduced populations of W. auropunctata contribute to its invasive success.     

The Evolution of Invasiveness in Garden Ants

It is unclear why some species become successful invaders whilst others fail, and whether invasive success depends on pre-adaptations already present in the native range or on characters evolving de-novo after introduction. Ants are among the worst invasive pests, with Lasius neglectus and its rapid spread through Europe and Asia as the most recent example of a pest ant that may become a global problem. Here, we present the first integrated study on behavior, morphology, population genetics, chemical recognition and parasite load of L. neglectus and its non-invasive sister species L. turcicus. We find that L. neglectus expresses the same supercolonial syndrome as other invasive ants, a social system that is characterized by mating without dispersal and large networks of cooperating nests rather than smaller mutually hostile colonies. We conclude that the invasive success of L. neglectus relies on a combination of parasite-release following introduction and pre-adaptations in mating system, body-size, queen number and recognition efficiency that evolved long before introduction. Our results challenge the notion that supercolonial organization is an inevitable consequence of low genetic variation for chemical recognition cues in small invasive founder populations. We infer that low variation and limited volatility in cuticular hydrocarbon profiles already existed in the native range in combination with low dispersal and a highly viscous population structure. Human transport to relatively disturbed urban areas thus became the decisive factor to induce parasite release, a well established general promoter of invasiveness in non-social animals and plants, but understudied in invasive social insects.     

Chemical mimicry of the ant Oecophylla smaragdina by the myrmecophilous spider Cosmophasis bitaeniata: Is it colony-specific?

Workers of most social insects can distinguish between nestmates and non-nestmates, and actively attack the latter if they attempt to intrude into the nest or surrounding territory. Nevertheless, there are many records of heterospecific organisms living within the nests of social insects, and they are thought to gain access through chemical mimicry. The salticid spider Cosmophasis bitaeniata lives within the leaf nests of the ant Oecophylla smaragdina, where it preys on the ant larvae. We investigated, using behavioural bioassays and chemical analyses, whether the previously reported resemblance of the cuticular hydrocarbons of ant and spider was colony-specific. Behavioural experiments revealed that the spiders can distinguish between nestmate and non-nestmate major workers and are less inclined to escape when confined with ants that are nestmates. More significantly, C. bitaeniata were more likely to capture ant larvae from nestmate minor workers than non-nestmate minor workers. The chemical analyses revealed that the cuticular hydrocarbon profiles of the spiders and the major workers of the ant colonies were colony-specific. However, the hydrocarbon profiles of C. bitaeniata do not match those of the major workers of O. smaragdina from the same colony. Perhaps the colony-specific cuticular hydrocarbon profiles of C. bitaeniata function to obtain prey from the minor workers rather than avoid eliciting aggression from the major workers.     

Characterization of cuticular hydrocarbons of diploid and haploid males, workers and queens of the stingless bee Melipona quadrifasciata

Males, queens and workers of stingless bees show differences in external morphology, behaviour and roles within a colony. In addition, each individual has a cuticular chemical signature responsible for mutual communication that is essential for maintaining the integrity of the colony. In this paper we characterize the cuticular hydrocarbon composition of newly emerged diploid and haploid males, workers and virgin queens of Melipona quadrifasciata by gas chromatography-mass spectrometry (GC/MS) analysis. This is the first time that the cuticular profile of diploid males in a species of stingless bee has been characterized. We found differences in the cuticular hydrocarbon composition among males, workers and virgin queens, recording both qualitative and quantitative differences among individuals of different phenotypes. However, no compound was found exclusively in diploid males. The cuticular chemical profiles of haploid and diploid males were very similar to those of workers. Moreover, the cuticular lipids of males and workers were significantly different from those of queens. Tricosane, pentacosene-2 and 7-methyl-heptacosane were the compounds responsible for this significant separation. This result correlates with the behavioural and morphological differences among these phenotypes.     

Under the volcano: phylogeography and evolution of the cave-dwelling Palmorchestia hypogaea (Amphipoda, Crustacea) at La Palma (Canary Islands)

Background

The invasive garden ant, Lasius neglectus, is the most recently detected pest ant and the first known invasive ant able to become established and thrive in the temperate regions of Eurasia. In this study, we aim to reconstruct the invasion history of this ant in Europe analysing 14 populations with three complementary approaches: genetic microsatellite analysis, chemical analysis of cuticular hydrocarbon profiles and behavioural observations of aggression behaviour. We evaluate the relative informative power of the three methodological approaches and estimate both the number of independent introduction events from a yet unknown native range somewhere in the Black Sea area, and the invasive potential of the existing introduced populations.

Results

Three clusters of genetically similar populations were detected, and all but one population had a similar chemical profile. Aggression between populations could be predicted from their genetic and chemical distance, and two major clusters of non-aggressive groups of populations were found. However, populations of L. neglectus did not separate into clear supercolonial associations, as is typical for other invasive ants.

Conclusion

The three methodological approaches gave consistent and complementary results. All joint evidence supports the inference that the 14 introduced populations of L. neglectus in Europe likely arose from only very few independent introductions from the native range, and that new infestations were typically started through introductions from other invasive populations. This indicates that existing introduced populations have a very high invasive potential when the ants are inadvertently spread by human transport.