Nest refuse of Acromyrmex balzani (Hymenoptera: Formicidae) increases the plant vigor in Turnera subulata (Turneraceae)

Some studies report the positive effect of organic residues from ant nests on soil properties and on the structure of the adjacent plant community in field experiments, but there is a gap about the effect on individual species. The purpose of the present study was to compare the soil nutrient content and the development of Turnera subulata Smith, an ornamental species, in the presence of the nest refuse (basically composed of fragments of grass leaves and the symbiotic fungus) produced by the leaf-cutting ant Acromyrmex balzani (Emery, 1890) or in control soil through a greenhouse pot experiment. The experiment was carried out with two treatments: control soil and soil with 25% of nest refuse. The plants were kept in 1L pots for 90 days. We evaluated the parameters: plant height, stem diameter, root length, number of leaves, dry weight of the root, dry and fresh aboveground biomass. Additionally, the relative chlorophyll content and leaf nutrients were used as nutritional parameters. As a result, plants that grew in the soil with nest refuse showed significant higher values of all parameters evaluated when compared to the control treatment (p < 0.001). We conclude that this biofertilizer contributed to the production of more vigorous plants, being able to act on the local dynamics of nutrients in the ecosystems where A. balzani occurs. As it is relatively abundant and easy to collect, the refuse of A. balzani has the potential to be used as an alternative substrate in the production of shortlife cycle plants.

Sperm Storage Costs Determine Survival and Immunocompetence in Newly Mated Queens of the Leaf-Cutting Ant Atta colombica

In the leaf-cutting ant Atta colombica, queens receive ejaculates from multiple males during one single mating event early in their lives. A queen’s fertility and fitness therefore depend on maximizing the number of sperm cells she can store and maintain inside her spermatheca. Previous studies implied significant physiological mating costs, either originating from energetic investments maximizing sperm survival, or from resolving sexual conflicts to terminate male-driven incapacitation of rival sperm via serine proteases found in seminal fluid. Here we conducted an artificial insemination experiment, which allowed us to distinguish between the effects of sperm and seminal fluid within the queen’s sexual tract on her survival and immunocompetence. We found significantly higher mortality in queens that we had inseminated with sperm, independently of whether seminal fluid was present or not. Additionally, after receiving sperm, heavier queens had a higher probability of survival compared to lightweight queens, and immunocompetence decreased disproportionally for queens that had lost weight during the experiment. These findings indicate that queens pay significant physiological costs for maintaining and storing sperm shortly after mating. On the other hand, the presence of seminal fluid within the queens’ sexual tract neither affected their survival nor their immunocompetence. This suggests that the energetic costs that queens incur shortly after mating are primarily due to investments in sperm maintenance and not costs of terminating conflicts between competing ejaculates. This outcome is consistent with the idea that sexually selected traits in social insects with permanent castes can evolve only when they do not affect survival or life-time fitness of queens in any significant way.

Decoupled evolution of mating biology and social structure in Acromyrmex leaf-cutting ants

Insect societies vary greatly in their social structure, mating biology, and life history. Polygyny, the presence of multiple reproductive queens in a single colony, and polyandry, multiple mating by females, both increase the genetic variability in colonies of eusocial organisms, resulting in potential reproductive conflicts. The co-occurrence of polygyny and polyandry in a single species is rarely observed across eusocial insects, and these traits have been found to be negatively correlated in ants. Acromyrmex leaf-cutting ants are well-suited for investigating the evolution of complex mating strategies because both polygyny and polyandry co-occur in this genus. We used microsatellite markers and parentage inference in five South American Acromyrmex species to study how different selective pressures influence the evolution of polygyny and polyandry. We show that Acromyrmex species exhibit independent variation in mating biology and social structure, and polygyny and polyandry are not necessarily negatively correlated within genera. One species, Acromyrmex lobicornis, displays a significantly lower mating frequency compared to others, while another species, A. lundii, appears to have reverted to obligate monogyny. These variations appear to have a small impact on average intra-colonial relatedness, although the biological significance of such a small effect size is unclear. All species show significant reproductive skew between patrilines, but there was no significant difference in reproductive skew between any of the sampled species. We find that the evolution of social structure and mating biology appear to follow independent evolutionary trajectories in different species. Finally, we discuss the evolutionary implications that mating biology and social structure have on life history evolution in Acromyrmex leaf-cutting ants. Significance statement Many species of eusocial insects have colonies with multiple queens (polygyny), or queens mating with multiple males (polyandry). Both behaviors generate potentially beneficial genetic diversity in ant colonies as well as reproductive conflict. The co-occurrence of both polygyny and polyandry in a single species is only known from few ant species. Leaf-cutting ants have both multi-queen colonies and multiply mated queens, providing a well-suited system for studying the co-evolutionary dynamics between mating behavior and genetic diversity in colonies of eusocial insects. We used microsatellite markers to infer the socio-reproductive behavior in five South American leaf-cutter ant species. We found that variation in genetic diversity in colonies was directly associated with the mating frequencies of queens, but not with the number of queens in a colony. We suggest that multi-queen nesting and mating frequency evolve independently of one another, indicating that behavioral and ecological factors other than genetic diversity contribute to the evolution of complex mating behaviors in leaf-cutting ants.

Effects of Trichoderma strigosellum in Eucalyptus urophylla Development and Leaf-Cutting Ant Behavior

Fungal endophytes can protect plants against herbivory and be used to control leaf-cutting ants. In this study, we aimed to evaluate the potential of endophytic colonization of Eucalyptus urophylla by three filamentous fungal species and their influence on the plant development and foraging behavior of Atta sexdens. The study design was completely randomized and comprised a factorial scheme of 4 × 3, three antagonistic fungal species (Escovopsis sp., Metarhizium anisopliae, and Trichoderma strigosellum) of the leaf-cutting ant, and one control and three inoculation methods (conidial suspension via foliar spray [FS] and soil drench [SD] inoculation, and seedlings inoculated with mycelium [SWM]). The SWM method allowed T. strigosellum to colonize all plant organs, and these plants exhibited higher height, leaf number, shoot dry mass, and total dry mass than the ones subjected to the other inoculation methods. The SWM method increased the plant height than the control plants and those inoculated with Escovopsis sp. and M. anisopliae. Trichoderma strigosellum, previously isolated from soil, colonized E. urophylla plants and positively influenced their development, as demonstrated by the SWM method. Trichoderma strigosellum promoted the increase in E. urophylla height compared with when the FS and SD methods were used (by 19.62% and 18.52%, respectively). Our results reveal that A. sexdens workers preferentially began cutting the leaves from plants not previously colonized by T. strigosellum. This behavior can be explained by modifications in the phenotypic traits of the eucalyptus leaves.

TREE SPECIES SUSCEPTIBILITY TO LEAF-CUTTING ANTS ATTACK IN CARBON NEUTRALIZATION PLANTATIONS

Greenhouse gas (GHG) emissions neutralize planting are one of the options for climate changes mitigating. Leaf-cutter ants attack is a threat to the plantations proper development. Ants have selective foraging, which makes it important to know this selectivity and, thus, choose more suitable species to neutralization planting compose. Thus, the goal of the present study was to evaluate the tree species susceptibility to be attacked by leaf-cutter ants in carbon neutralization plantations. The study was carried out in mixed plantations in Viçosa-MG and a classification was created for the present species. The Damage Index (DI) was created by multiplying the Mean of Severity (MS) and the Frequency of Attacks (FA). The species were classified according to the indication for neutralization plantations (indicated, moderately indicated, and not indicated) and potential species for the extraction of natural insecticides. From the 59 species evaluated, 22 were classified as suitable for neutralization plantations, 6 as moderately indicated, 24 as not indicated, and 7 as potential species for extracting natural insecticides. Keywords: forest carbon; pest control; carbon offset; mixed plantings.

Landscape genetics across the Andes Mountains: Environmental variation drives genetic divergence in the leaf-cutting ant Atta cephalotes

Disentangling the mechanisms underlying spatial distribution of genetic variation into those due to environment or physical barriers from mere geographic distance is challenging in complex landscapes. The Andean uplift represents one of the most heterogeneous habitats where these questions remain unexplored as multiple mechanisms might interact, confounding their relative roles. We explore this broad question in the leaf-cutting ant Atta cephalotes, a species distributed across the Andes Mountains using nuclear microsatellite markers and mtCOI gene sequences. We investigate spatial genetic divergence across the Western range of Northern Andes in Colombia by testing the relative role of alternative scenarios of population divergence, including geographic distance (IBD), climatic conditions (IBE), and physical barriers due to the Andes Mountains (IBB). Our results reveal substantial genetic differentiation among A. cephalotes populations for both types of markers, but only nuclear divergence followed a hierarchical pattern with multiple models of genetic divergence imposed by the Western range. Model selection showed that IBD, IBE (temperature and precipitation), and IBB (Andes mountains) models, often proposed as individual drivers of genetic divergence, interact, and explain up to 33% of genetic divergence in A. cephalotes. IBE models remained significant after accounting for IBD, suggesting that environmental factors play a more prominent role when compared to IBB. These factors in combination with idiosyncratic dispersal patterns of ants appear to determine hierarchical patterns of gene flow. This study enriches our understanding of the forces shaping population divergence in complex habitat landscapes.

Leaf-cutting ants’ critical and voluntary thermal limits show complex responses to size, heating rates, hydration level, and humidity

Thermal variation has complex effects on organisms and they respond to these effects through combined behavioral and physiological mechanisms. However, it is less clear how these traits combine in response to changes in body condition (e.g., size, hydration) and environmental factors that surround the heating process (e.g., relative humidity, start temperatures, heating rates). We tested whether these body conditions and environmental factors influence sequentially measured Voluntary Thermal Maxima (VTmax) and Critical Thermal Maxima, (CTmax) in leaf-cutting ants (Atta sexdens rubropilosa, Forel, 1908). VTmax and CTmax reacted differently to changes in body size and relative humidity, but exhibited similar responses to hydration level, start temperature, and heating rate. Strikingly, the VTmax of average-sized workers was closer to their CTmax than the VTmax of their smaller and bigger sisters, suggesting foragers maintain normal behavior at higher temperatures than sister ants that usually perform tasks within the colony. Previous experiments based on hot plate designs might overestimate ants’ CTmax. VTmax and CTmax may respond concomitantly or not to temperature rises, depending on body condition and environmental factors.