Interactions between seed-dispersing ant species affect plant community composition in field mesocosms.

In generalized mutualisms, species vary in the quality of services they provide to their partners directly via traits that affect partner fitness and indirectly via traits that influence interactions among mutualist species that play similar functional roles. Myrmecochory, or seed dispersal by ants, is a generalized mutualism with ant species varying in the quality of dispersal services they provide to their plant partners. Variation in ant species identity can directly impact seed dispersal patterns and plant community composition; however, we know less about how interactions among seed-dispersing ant species indirectly influence plant partners. The invasive ant Myrmica rubra, is a high-quality seed-disperser in its native range that interacts with myrmecochores (ant-dispersed plants) and the high-quality seed disperser Aphaenogaster sp. in its invaded range. We use this system to examine how interactions between two functionally similar mutualist ant species influence the recruitment and community composition of ant-dispersed plants. We performed a field mesocosm experiment and a laboratory behavioural experiment to compare discovery and dominance behaviours between ant species, and seed dispersal and seedling recruitment of four myrmecochore species among intraspecific interaction treatments of each ant species and an interspecific interaction treatment. We found that M. rubra was better at discovering and dispersing seeds, but Aphaenogaster sp. was dominantly aggressive over M. rubra. Interspecific interactions dampened seed dispersal relative to dispersal by the better disperser. Despite this dampening, we found no effect of interspecific interactions on seedling recruitment. However, community composition of seedlings in the interspecific interaction treatment was more similar to composition in the aggressively dominant ant (Aphaenogaster sp.) treatment than in the better discoverer ant M. rubra treatment. We show that interspecific interactions between mutualist species in the same functional guild affect the outcome of mutualistic interactions with partner species. Despite the native ant dispersing fewer seeds, its dominance over the subordinate (invasive) ant has the potential to allow for some level of biotic resistance against the effects of M. rubra on plant communities when these species coexist.

Invasive ants disperse seeds farther than native ants, affecting the spatial pattern of seedling recruitment and survival.

Mutualists can vary in the quantity and quality of service which they provide to their partners. Variation in seed disperser quality depends on seed-processing traits, dispersal distance, and deposition location, all of which ultimately affect plant fitness. Here, we compared these aspects of seed dispersal quality between a native and an invasive ant species, and examined how they affect competition and plant performance. Using experimental mesocosm communities, we examined how these two ant species affect the spatial pattern of recruitment and establishment for four myrmecochorous plant species, including one invasive species. We measured the locations of dispersed seedlings relative to ant nests, adult plants, and other dispersed seedlings, as well as measured the effects of location on plant performance. The invasive ant, Myrmica rubra, secondarily dispersed seeds farther from its nests, creating a less clumped pattern of seedling recruitment compared to the native ant, Aphaenogaster rudis. Plant species responded differently to dispersal. Invasive seedlings recruited farther from adult plants than native seedlings, and had higher survival the farther they were from conspecifics. In contrast, native plants had higher survival and grew taller when dispersed farther from invasive plants. We show that seed-dispersing ant partners differ in mutualist quality creating differences in dispersal distance and deposition location that affects a plant's competitive environment. Our results reveal the potential for long-term consequences on plant community structure with changing ant partner identity. We emphasize the need to examine dispersal quality in addition to quantity to uncover the importance of partner identity in structuring communities.

Patterns and dynamics of neutral lipid fatty acids in ants - implications for ecological studies.

BACKGROUND: Trophic interactions are a fundamental aspect of ecosystem functioning, but often difficult to observe directly. Several indirect techniques, such as fatty acid analysis, were developed to assess these interactions. Fatty acid profiles may indicate dietary differences, while individual fatty acids can be used as biomarkers. Ants are among the most important terrestrial animal groups, but little is known about their lipid metabolism, and no study so far used fatty acids to study their trophic ecology. We set up a feeding experiment with high- and low-fat food to elucidate patterns and dynamics of neutral lipid fatty acids (NLFAs) assimilation in ants. We asked whether dietary fatty acids are assimilated through direct trophic transfer, how diet influences NLFA total amounts and patterns over time, and whether these assimilation processes are similar across species and life stages. RESULTS: Ants fed with high-fat food quickly accumulated specific dietary fatty acids (C18:2n6, C18:3n3 and C18:3n6), compared to ants fed with low-fat food. Dietary fat content did not affect total body fat of workers or amounts of fatty acids extensively biosynthesized by animals (C16:0, C18:0, C18:1n9). Larval development had a strong effect on the composition and amounts of C16:0, C18:0 and C18:1n9. NLFA compositions reflected dietary differences, which became more pronounced over time. Assimilation of specific dietary NLFAs was similar regardless of species or life stage, but these factors affected dynamics of other NLFAs, composition and total fat. CONCLUSIONS: We showed that ants accumulated certain dietary fatty acids via direct trophic transfer. Fat content of the diet had no effect on lipids stored by ants, which were able to synthesize high amounts of NLFAs from a sugar-based diet. Nevertheless, dietary NLFAs had a strong effect on metabolic dynamics and profiles. Fatty acids are a useful tool to study trophic biology of ants, and could be applied in an ecological context, although factors that affect NLFA patterns should be taken into account. Further studies should address which NLFAs can be used as biomarkers in natural ant communities, and how factors other than diet affect fatty acid dynamics and composition of species with distinct life histories.

Keep the nest clean: survival advantages of corpse removal in ants.

Sociality increases exposure to pathogens. Therefore, social insects have developed a wide range of behavioural defences, known as 'social immunity'. However, the benefits of these behaviours in terms of colony survival have been scarcely investigated. We tested the survival advantage of prophylaxis, i.e. corpse removal, in ants. Over 50 days, we compared the survival of ants in colonies that were free to remove corpses with those that were restricted in their corpse removal. From Day 8 onwards, the survival of adult workers was significantly higher in colonies that were allowed to remove corpses normally. Overall, larvae survived better than adults, but were slightly affected by the presence of corpses in the nest. When removal was restricted, ants removed as many corpses as they could and moved the remaining corpses away from brood, typically to the nest corners. These results show the importance of nest maintenance and prophylactic behaviour in social insects.

Increase in quantity and quality of suitable areas for invasive species as climate changes.

As climatically suitable range projections become increasingly used to assess distributions of species, we recommend systematic assessments of the quality of habitat in addition to the classical binary classification of habitat. We devised a method to assess occurrence probability, captured by a climatic suitability index, through which we could determine variations in the quality of potential habitat. This relative risk assessment circumvents the use of an arbitrary suitability threshold. We illustrated our method with 2 case studies on invasive ant species. We estimated invasion potential of the destroyer ant (Monomorium destructor) and the European fire ant (Myrmica rubra) on a global scale currently and by 2080 with climate change. We found that 21.1% of the world's landmass currently has a suitable climate for the destroyer ant and 16% has a suitable climate for European fire ant. Our climatic suitability index showed that both ant species would benefit from climate change, but in different ways. The size of the potential distribution increased by 35.8% for the destroyer ant. Meanwhile, the total area of potential distribution remained the same for the European fire ant (>0.05%), but the level of climatic suitability within this range increased greatly and led to an improvement in habitat quality (i.e., of invasive species' establishment likelihood). Either through quantity or quality of suitable areas, both invasive ant species are likely to increase the extent of their invasion in the future, following global climate change. Our results show that species may increase their range if either more areas become suitable or if the available areas present improved suitability. Studies in which an arbitrary suitability threshold was used may overlook changes in area quality within climatically suitable areas and as a result reach incorrect predictions. Incremento de la Cantidad y Calidad de Áreas Idóneas para Especies Invasoras a Medida que Cambia el Clima.

Know your foe: synanthropic spiders are deterred by semiochemicals of European fire ants.

Many ants prey on spiders, suggesting that web-building spiders may avoid micro-locations near ant colonies or frequented by foraging ants. Here we tested the hypothesis that ant-derived semiochemicals deter synanthropic spiders. To generate stimuli, we exposed filter paper for 12 h to workers of European fire ants, Myrmica rubra, black garden ants, Lasius niger, or western carpenter ants, Camponotus modoc, and then offered select urban spiders in three-chamber olfactometer bioassays a choice between ant-exposed filter paper and unexposed control filter paper. Semiochemical deposits of M. rubra, but not of L. niger or C. modoc, had a significant deterrent effect on subadults of the false black widow, Steatoda grossa, the black widow, Latrodectus hesperus, and the hobo spider, Eratigena agrestis, as well as a moderate (but statistically not significant) deterrent effect on the cross spider, Araneus diadematus. The deterrent effect caused by semiochemical deposits of M. rubra may be attributable to the aggressive nature and efficient foraging of M. rubra in its invaded North American range, exerting selection pressure on community members to recognize M. rubra semiochemicals and to avoid micro-locations occupied by M. rubra.

Prophylactic Avoidance of Hazardous Prey by the Ant Host Myrmica rubra.

Ants are the hosts of many microorganisms, including pathogens that are incidentally brought inside the nest by foragers. This is particularly true for scavenging species, which collect hazardous food such as dead insects. Foragers limit sanitary risks by not retrieving highly infectious prey releasing entomopathogenic fungal spores. This study investigates whether similar prophylactic strategies are also developed for food associated with weak or delayed risks of fungal contamination. We compared, in Myrmica rubra ant colonies, the retrieval dynamics of dead flies that were (1) conidia-free, (2) covered with a low amount of Metarhizium brunneum entomopathogenic conidia or (3) recently fungus-killed but not yet sporulating. Foragers mostly avoided fungus-killed prey and delayed the retrieval of conidia-covered flies. A second sanitary filter occurred inside the nest through a careful inspection of the retrieved prey. Ultimately, ants mostly consumed conidia-free and conidia-covered flies, but they relocated and discarded all fungus-killed prey outside of the nest. Our study confirms that, as a host of generalist entomopathogenic fungi, Myrmica rubra ants have developed a prophylactic avoidance and a differential management of prey depending on their infectious potential. We discuss the functional value as well as the possible cues underlying pathogen avoidance and prey discrimination in ants.

Sublethal insecticide exposure of an herbivore alters the response of its predator.

Sublethal insecticide exposure poses risks for many non-target organisms and is a challenge for successful implementation of integrated pest management (IPM) programs. Next to detrimental effects of short-term insecticide exposure on fitness-related traits of organisms, also properties such as chemical signaling traits can be altered, which mediate intra- and interspecific communication. We investigated the effects of different durations of larval sublethal exposure to the pyrethroid lambda-cyhalothrin on performance traits of larvae and adults of the herbivorous mustard leaf beetle, Phaedon cochleariae. Moreover, by applying a direct contact and olfactometer bioassays, we determined the reaction of a generalist predator, the ant Myrmica rubra, towards insecticide-exposed and unexposed herbivore larvae and their secretions. Already short-term sublethal insecticide exposure of a few days caused a prolonged larval development and a reduced adult body mass of males. These effects may result from an insecticide-induced reduction in energy reserves. Furthermore, ants responded more frequently to insecticide-exposed than to unexposed larvae of P. cochleariae and their secretions. This increased responsiveness of ants towards insecticide-exposed larvae may be due to an insecticide-induced change in synthesis of chrysomelidial and epichrysomelidial, the dominant compounds of the larval secretion, which act defensive against various generalist predators. In conclusion, the results highlight that short-term insecticide exposure can impair the fitness of an herbivorous species due to both direct toxic effects and an increased responsiveness of predators. Consequently, exposure of single non-target species can have consequences for ecological communities in both natural habitats and IPM programs.

Foodborne Transmission of Deformed Wing Virus to Ants (Myrmica rubra).

Virus host shifts occur frequently, but the whole range of host species and the actual transmission pathways are often poorly understood. Deformed wing virus (DWV), an RNA virus described from honeybees (Apis mellifera), has been shown to have a broad host range. Since ants are often scavenging on dead honeybees, foodborne transmission of these viruses may occur. However, the role of the ant Myrmica rubra as an alternative host is not known and foodborne transmission to ants has not been experimentally addressed yet. Here, we show with a 16-week feeding experiment that foodborne transmission enables DWV type-A and -B to infect M. rubra and that these ants may serve as a virus reservoir. However, the titers of both plus- and minus-sense viral RNA strands decreased over time. Since the ants were fed with highly virus-saturated honeybee pupae, this probably resulted in initial viral peaks, then approaching lower equilibrium titers in infected individuals later. Since DWV infections were also found in untreated field-collected M. rubra colonies, our results support the wide host range of DWV and further suggest foodborne transmission as a so far underestimated spread mechanism.

Identification and Functional Characterization of a Novel Insecticidal Decapeptide from the Myrmicine Ant Manica rubida.

Ant venoms contain many small, linear peptides, an untapped source of bioactive peptide toxins. The control of agricultural insect pests currently depends primarily on chemical insecticides, but their intensive use damages the environment and human health, and encourages the emergence of resistant pest populations. This has promoted interest in animal venoms as a source of alternative, environmentally-friendly bio-insecticides. We tested the crude venom of the predatory ant, Manica rubida, and observed severe fitness costs in the parthenogenetic pea aphid (Acyrthosiphon pisum), a common agricultural pest. Therefore, we explored the M. rubida venom peptidome and identified a novel decapeptide U-MYRTX-MANr1 (NH2-IDPKVLESLV-CONH2) using a combination of Edman degradation and de novo peptide sequencing. Although this myrmicitoxin was inactive against bacteria and fungi, it reduced aphid survival and reproduction. Furthermore, both crude venom and U-MYRTX-MANr1 reversibly paralyzed injected aphids and induced a loss of body fluids. Components of M. rubida venom may act on various biological targets including ion channels and hemolymph coagulation proteins, as previously shown for other ant venom toxins. The remarkable insecticidal activity of M. rubida venom suggests it may be a promising source of additional bio-insecticide leads.

Ants Sense, and Follow, Trail Pheromones of Ant Community Members.

Ants deposit trail pheromones that guide nestmates to food sources. We tested the hypotheses that ant community members (Western carpenter ants, Camponotus modoc; black garden ants, Lasius niger; European fire ants, Myrmica rubra) (1) sense, and follow, each other's trail pheromones, and (2) fail to recognize trail pheromones of allopatric ants (pavement ants, Tetramorium caespitum; desert harvester ants, Novomessor albisetosus; Argentine ants, Linepithema humilis). In gas chromatographic-electroantennographic detection analyses of a six-species synthetic trail pheromone blend (6-TPB), La. niger, Ca. modoc, and M. rubra sensed the trail pheromones of all community members and unexpectedly that of T. caespitum. Except for La. niger, all species did not recognize the trail pheromones of N. albisetosus and Li. humilis. In bioassays, La. niger workers followed the 6-TPB trail for longer distances than their own trail pheromone, indicating an additive effect of con- and hetero-specific pheromones on trail-following. Moreover, Ca. modoc workers followed the 6-TPB and their own trail pheromones for similar distances, indicating no adverse effects of heterospecific pheromones on trail-following. Our data show that ant community members eavesdrop on each other's trail pheromones, and that multiple pheromones can be combined in a lure that guides multiple species of pest ants to lethal food baits.

Emergency measures: Adaptive response to pathogen intrusion in the ant nest.

Ants have developed prophylactic and hygienic behaviours in order to limit risks of pathogenic outbreaks inside their nest, which are often called social immunity. Here, we test whether ants can adapt the "social immune response" to the level of pathogenic risk in the colony. We challenged Myrmica rubra colonies with dead nestmates that had either died from being frozen or from infection by the fungus Metarhizium anisopliae. Ant survival was compromised by the presence of the fungus-bearing corpses: workers died faster with a significantly lower survival from the 4th day compared to workers challenged with freeze-killed corpses. When faced with fungus-bearing corpses, workers responded quickly by increasing hygienic behaviours: they spent more time cleaning the nest, moving the corpses, and self-grooming. Ants in fungus-threatened colonies also decreased contact rates with other workers, and moved corpses further in the corners of the nest than in colonies in contact with non-infected corpses. These results show that ant colonies are able to assess the risk level associated with the presence of corpses in the nest, and adjust their investment in terms of hygienic behaviour.

Unexpected reactions of a generalist predator towards defensive devices of cassidine larvae (Coleoptera, Chrysomelidae).

Studies of multitrophic interactions show that insect faeces may act as a defensive device against predators, as kairomone source for attraction of antagonists and as a significant energy source for micro-organisms. In the present study, we investigated effects of larval faeces from leaf beetles of the subfamiliy Cassidinae towards a generalist predator, the ant Myrmica rubra. Most cassidine larvae collect their faeces together with exuviae as so-called abdominal defensive shields on two movable spines at the posterior tip. The effects of these abdominal shields towards M. rubra were studied in three cassidine species, which feed mono- or oligophagously upon tansy (Chrysanthemum vulgare): Cassida denticollis and C. sanguinosa which possess such faecal shields and, for comparison, C. stigmatica with shields made of exuviae only (=skin shield). Bioassays revealed that larvae with both faecal and skin shields were attacked by the ant M. rubra more often than larvae whose shields had been removed. This attractiveness of shields towards ants contrasts with other studies, which found that abdominal shields of chrysomelid larvae act as defensive mechanisms against generalist predators like ants. To characterize the attractive cues of the shields, we studied possible chemical and physical stimuli. Olfactometer bioassays with M. rubra and chemical analyses revealed that plant-derived volatiles from faecal shields of C. denticollis attracted the ant, whereas odour from skin shields of C. stigmatica did not. Skin shields also emitted volatiles which derived from tansy, but in much lower quantities. Exclusion of contact to surface chemicals of a faecal shield reduced the ants' aggressive behaviour, whereas a change in the moisture content of a faecal shield had no influence. Visual stimuli cannot be ruled out as enhancing the ants' reaction towards faecal shields with their attractive volatiles, and are suggested to play a major role in the ants' response towards skin shields. This novel attractive effect of the abdominal shields of cassidine larvae is discussed, especially with respect to host plant chemistry and possible functions of the shields that might outweigh the negative consequences of the attraction of the predator M. rubra.