Competition-based screening helps to secure the evolutionary stability of a defensive microbiome.

BACKGROUND: The cuticular microbiomes of Acromyrmex leaf-cutting ants pose a conundrum in microbiome biology because they are freely colonisable, and yet the prevalence of the vertically transmitted bacteria Pseudonocardia, which contributes to the control of Escovopsis fungus garden disease, is never compromised by the secondary acquisition of other bacterial strains. Game theory suggests that competition-based screening can allow the selective recruitment of antibiotic-producing bacteria from the environment, by providing abundant resources to foment interference competition between bacterial species and by using Pseudonocardia to bias the outcome of competition in favour of antibiotic producers. RESULTS: Here, we use RNA-stable isotope probing (RNA-SIP) to confirm that Acromyrmex ants can maintain a range of microbial symbionts on their cuticle by supplying public resources. We then used RNA sequencing, bioassays, and competition experiments to show that vertically transmitted Pseudonocardia strains produce antibacterials that differentially reduce the growth rates of other microbes, ultimately biassing the bacterial competition to allow the selective establishment of secondary antibiotic-producing strains while excluding non-antibiotic-producing strains that would parasitise the symbiosis. CONCLUSIONS: Our findings are consistent with the hypothesis that competition-based screening is a plausible mechanism for maintaining the integrity of the co-adapted mutualism between the leaf-cutting ant farming symbiosis and its defensive microbiome. Our results have broader implications for explaining the stability of other complex symbioses involving horizontal acquisition.

Underground anemotactic orientation in leaf-cutting ants: perception of airflow and experience-dependent choice of airflow direction during digging.

Air exchange between the large nests of Atta vollenweideri leaf-cutting ants and the environment strongly relies on a passive, wind-induced ventilation mechanism. Air moves through nest tunnels and airflow direction depends on the location of the tunnel openings on the nest mound. We hypothesized that ants might use the direction of airflow along nest tunnels as orientation cue in the context of climate control, as digging workers might prefer to broaden or to close tunnels with inflowing or outflowing air in order to regulate nest ventilation. To investigate anemotactic orientation in Atta vollenweideri, we first tested the ants' ability to perceive air movements by confronting single workers with airflow stimuli in the range 0 to 20 cm/s. Workers responded to airflow velocities ≥ 2 cm/s, and the number of ants reacting to the stimulus increased with increasing airflow speed. Second, we asked whether digging workers use airflow direction as an orientation cue. Workers were exposed to either inflow or outflow of air while digging in the nest and could subsequently choose between two digging sites providing either inflow or outflow of air, respectively. Workers significantly chose the side with the same airflow direction they experienced before. When no airflow was present during initial digging, workers showed no preference for airflow directions. Workers developed preferences for airflow direction only after previous exposure to a given airflow direction. We suggest that experience-modified anemotaxis might help leaf-cutting ants spatially organize their digging activity inside the nest during tasks related to climate control.

Reproductive ecology of phorid parasitoids in relation to the head size of leaf-cutting ants Atta sexdens Forel.

The leaf-cutting ant Atta sexdens Forel (Hymenoptera: Formicidae) is one of the most damaging agricultural pests in the Neotropics. Management strategies predominantly rely on the use of general insecticides. What is needed are more species-specific and environmentally friendly options. Parasitioids such as phorid flies (Diptera: Phoridae) may be one such option, but a greater understanding of the ecology of the flies and their ant hosts is essential to devise biological control strategies. Here we report parasitism rates, ant host size, parasitoid abundance per host and resultant sex ratios of two phorid species Apocephalus attophilus Borgmeier and Eibesfeldtphora tonhascai Brown parasitizing A.sexdens. The two species achieved parasitism rates of 1.48 and 1.46%, respectively and the pupal period was 14.7 ± 1.1 days and 22.1 ± 2.8 days, respectively. There was no significant difference between the head capsule width of ants parasitized by either A. attophilus or E. tonhascai. Likewise, there was no significant effect between the head capsule width of parasitized and unparasitized ants for both species. A significant positive correlation was found between the head capsule width of the parasitized ants and the number of adult parasitoids A. attophilus emerged. Ants parasitized by E. tonhascai survived significantly longer than those parasitized by A. attophilus. There was no significant effect of ant head width on the sex ratio of the offspring of either parasitoid species and no significant difference in the sex ratio (male: female) of their offspring. In summary, these data addressed here are important steps when considering natural enemies for biological control. Studying survival of the parasitized ants, parasitoid offspring sex ratio and host size preference allows for a better understanding of ant natural biological control in the field and can help in rearing of A. attophilus and E. tonhascai in laboratory.

Antagonism of Trichoderma isolates against Leucoagaricus gongylophorus (Singer) Möller.

Filamentous fungi from the genus Trichoderma are commonly found in soil. They are considered facultative mycoparasites, and are antagonists of other fungi such as the cultivar of leaf-cutting ants (Leucoagaricus gongylophorus). The aim of the present study was to bioprospect Trichoderma spp. from different soils collected from Gurupi, Tocantins, Brazil, for antagonistic effects against the mutualistic fungus of leaf-cutting ants. To isolate filamentous fungi, samples were collected from six locations. Preliminarily, isolates were identified by morphological analysis as belonging to Trichoderma. Trichoderma spp. had their internal transcribed spacer region (ITS) of ribosomal RNA genes (rRNA) sequenced to confirm species-level taxonomy. L. gongylophorus was isolated from a laboratory ant colony. Antagonistic properties of seven isolates of Trichoderma against L. gongylophorus were measured using paired disks in Petri dishes with potato dextrose agar medium (PDA). All Trichoderma isolates inhibited the growth of L. gongylophorus in Petri dishes. Isolate 2 of Trichoderma spirale group exhibited slow mycelial growth in the Petri dish, and a high rate of inhibition against L. gongylophorus. This isolate is a promising fungus for field tests of biological control methods for leaf-cutting ants.

Riding with the ants.

Isolates of Teratosphaeriaceae have frequently been found in the integument of attine ants, proving to be common and diverse in this microenvironment. The LSU phylogeny of the ant-isolated strains studied revealed that they cluster in two main lineages. The first was associated with the genus Xenopenidiella whereas the other represented two ant-isolated lineages sister to the taxa Penidiella aggregata and P. drakensbergensis, which are allocated to the new genus Penidiellomyces. The genus Penidiella is limited to the lineage containing P. columbiana, which is not congeneric with Penidiellomyces or Penidiellopsis, nor with Simplicidiella, a novel genus introduced here to accommodate a strain isolated from ants. For species level analysis, the final 26 aligned sequences of the ITS (498 characters), cmdA (389 characters), tef1 (342 characters) and tub2 (446 characters) gene regions lead to the introduction of six new species in Xenopenidiella, and one in respectively Penidiellopsis and Simplicidiella. The species described in this study were distinguished by the combination of morphological and phylogenetic data. Novelties on the integument of leaf-cutting ants from Brazil include: Penidiellopsis ramosus, Xenopenidiella clavata, X. formica, X. inflata, X. laevigata, X. nigrescens, X. tarda spp. nov., and Simplicidiella nigra gen. & sp. nov. Beta-tubulin is recommended as primary barcode for the distinction of species in Penidiellopsis, whereas ITS was sufficient to distinguish species of Xenopenidiella.

Ant mediated redistribution of a xyloglucanase enzyme in fungus gardens of Acromyrmex echinatior.

BACKGROUND: Xyloglucan is an important component in plant cell walls that herbivores cannot digest without microbial symbionts. Leaf-cutting ants are major insect herbivores in the Neo-Tropics that rely on fungus-garden enzymes for degrading plant cell walls. However, many of these ants discard much of their harvested plant material after partial degradation, which has led to the hypothesis that the fungal symbionts are primarily producing cell wall degrading enzymes to gain access to intracellular nutrients rather than for obtaining sugars from recalcitrant cell wall polymers, such as (hemi-)cellulose. RESULTS: The fungal symbiont provides a single xyloglucanase (Xeg1) to its ant farmers by upregulating the expression of this protein in the inflated hyphal tips (gongylidia) that the ants ingest. Similar to other enzymes ingested this way, also Xeg1 is not digested but vectored to the fresh leaf-fragment pulp at the top of fungus gardens via ant fecal fluid. Xeg1 is 4-5 times more active in fecal fluid when ants ingest their normal fungal food, compared to a sucrose control diet, as expected when they cannot produce Xeg1 themselves. We confirm substrate specificity of fungal Xeg1 towards xyloglucan by heterologous expression in yeast and show that xyloglucanase activity is higher in the oldest, bottom layers of fungus gardens and in discarded debris material than in the upper and middle layers of fungus gardens. CONCLUSION: Our results are consistent with Xeg1 playing a role in the initial breakdown of plant cell wall hemicellulose to provide sugars for aggressive hyphal growth before intracellular proteins become available. Xeg1 does not play a major decomposition role in the middle layer of fungus gardens where it is produced by the gongylidia. Overall high xyloglucanase activity in old mycelium that is (about to be) discarded is striking and quite possibly serves defensive purposes by precluding that competing microorganisms can grow. Our results support the hypothesis that the ant-fungus symbiosis prioritizes access to the protein-rich contents of live plant cells and that carbohydrates are not a limiting resource.

Chemical disguise of myrmecophilous cockroaches and its implications for understanding nestmate recognition mechanisms in leaf-cutting ants.

BACKGROUND: Cockroaches of the genus Attaphila regularly occur in leaf-cutting ant colonies. The ants farm a fungus that the cockroaches also appear to feed on. Cockroaches disperse between colonies horizontally (via foraging trails) and vertically (attached to queens on their mating flights). We analysed the chemical strategies used by the cockroaches to integrate into colonies of Atta colombica and Acromyrmex octospinosus. Analysing cockroaches from nests of two host species further allowed us to test the hypothesis that nestmate recognition is based on an asymmetric mechanism. Specifically, we test the U-present nestmate recognition model, which assumes that detection of undesirable cues (non-nestmate specific substances) leads to strong rejection of the cue-bearers, while absence of desirable cues (nestmate-specific substances) does not necessarily trigger aggression. RESULTS: We found that nests of Atta and Acromyrmex contained cockroaches of two different and not yet described Attaphila species. The cockroaches share the cuticular chemical substances of their specific host species and copy their host nest's colony-specific cuticular profile. Indeed, the cockroaches are accepted by nestmate but attacked by non-nestmate ant workers. Cockroaches from Acromyrmex colonies bear a lower concentration of cuticular substances and are less likely to be attacked by non-nestmate ants than cockroaches from Atta colonies. CONCLUSIONS: Nest-specific recognition of Attaphila cockroaches by host workers in combination with nest-specific cuticular chemical profiles suggest that the cockroaches mimic their host's recognition labels, either by synthesizing nest-specific substances or by substance transfer from ants. Our finding that the cockroach species with lower concentration of cuticular substances receives less aggression by both host species fully supports the U-present nestmate recognition model. Leaf-cutting ant nestmate recognition is thus asymmetric, responding more strongly to differences than to similarities.

Herbivory by Atta vollenweideri: Reviewing the significance of grass-cutting ants as a pest of livestock.

The grass-cutting ant Atta vollenweideri is well suited for studies examining the negative effect leaf-cutting ants have on livestock production in South American grasslands because they forage on the same plants as cattle. This study investigated the impact of A. vollenweideri on livestock production in Argentinean rangelands. First, we assessed A. vollenweideri herbivory rates and its economic injury level (EIL). Second, using satellite imagery in a region covering 15,000 ha, we estimated the percentage of this area that surpassed the calculated EIL. Results showed that A. vollenweideri consumed approximately 276 kg of dry plant weight/ha/year, foraging mostly on grasses (70%). Additionally, ants cut 25% of herbs and 5% of trees. In summer and autumn, ants consumed more grasses, while in winter and spring, herbs and trees were also significantly cut. Ants consumed 7% of the forage demand needed to raise a calf according to the management regime applied by farmers. Our calculated EIL (5.85 nests/ha) falls in the range of previous studies. Colonies were absent in 93.6% of the surveyed area, while their density was below the EIL in 6.2% of the area. A. vollenweideri populations surpassed the EIL in only 0.2% of the area, which corresponds to 2.6% of the locations holding colonies. These results question the perception that Atta leaf-cutting ants are a pest of livestock production. Although ants consume a small percentage of cattle's forage demand, evidence that ants and cattle are competing in the few cases in which density surpasses the EIL is arguable. First, grass-cutting ants are capable of consuming herbs and trees in addition to the grasses on which cattle mostly feed. Second, there is no evidence indicating that both are cutting the same plant portions when preferences overlap. Third, evidence suggests that ants are not displaced under high-pressure grazing regimes by cattle. In the countries where A. vollenweideri is present, decision makers have promulgated several acts making its control mandatory. It is time to revisit the pest status of A. vollenweideri and include the use of EIL as a control criterion.

Chemical control of Acromyrmex lundi (Hymenoptera: Formicidae) in anthropic ecosystems.

BACKGROUND: Leaf-cutting ants are pests in agriculture, forests and pasture and, usually, controlled with toxic baits. The management and control of leaf-cutting ants is complex as a consequence of its sociability and behavior. We tested three toxic baits with abamectin, fipronil and sulfluramid to control colonies of Acromyrmex lundi Mayr, 1865 (Hymenoptera: Formicidae), in two seasons: 'Spring 2020' and 'Summer 2020/21', in Argentina. RESULTS: The percentage of effective bait carried in the spring was similar between treatments with 80%, 80% and 90% for those with abamectin, fipronil and sulfluramid. The ant flow of A. lundi decreased after toxic bait applications compared to the control through time in all treatments. This activity was higher in the spring than in the summer. The baits reduced the foraging activity of A. lundi from 5 to 96 days after application (DAA). Abamectin and fipronil baits controlled 50% and 67% of the nests in the spring, and 20% and 43% at 96 DAA in the summer. Sulfluramid was the best treatment, killing 90% of A. lundi nests. CONCLUSION: The baits decrease the foraging activity of A. lundi until 96 DAA, with total or partial death of its colonies varying between treatments. The ant symptoms with the sulfluramid bait appeared faster than with fipronil and may be a result of the low level of carry-over or differences in the chemical quality of the active ingredients. © 2022 Society of Chemical Industry.

Control of Amazonian Leaf-Cutting Ants (Hymenoptera: Formicidae): A Multi-criteria Analysis.

Leaf-cutting ants (Hymenoptera: Formicidae) are one of the main pests found in the Americas and they cause global economic losses worth several billions of dollars. While pesticides have been the most widely used control method, new management alternatives in a context of agroecological transition are now being considered. This study focuses on the leaf-cutting ants species found in the pan-Amazon region. As part of efforts to improve management of these pests, this multi-criteria analysis of control strategies covers a total of 691 experiments collected from 153 studies, and the control were evaluated as a function of their management efficacy, environmental and human health impacts, and their ease of application. Chemical control methods were effective but posed a danger to human health and the environment, whereas mechanical methods and integrated management were more sustainable but not always very effective. Some of the biocontrol methods were evaluated as effective and safe for the environment and human health, including the use of entomopathogenic fungi Beauveria bassiana (Bals.-Criv) Vuill. (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (Metschn.) Sorokïn (Hypocreales: Clavicipitaceae) in the form of bait or sprayed in the nest, or the application of plant mulch in the nest using Tithonia diversifolia (Hemsley) A. Gray (Asterales: Asteraceae) or Canavalia ensiformis L. DC. (Fabales: Fabaceae). Because of variations in the efficacy data between laboratory and field tests, we are in favor of evaluating these control methods during field studies with different leaf-cutting ant species and under different environmental conditions. These methods should adopt experimental arrangements that are appropriate for local socioeconomic conditions adapted for farmers.

Push-pull to manage leaf-cutting ants: an effective strategy in forestry plantations.

BACKGROUND: Leaf-cutting ants (LCAs) are amongst the most important forestry pests in South America. Currently, their control is carried out almost exclusively through the application of toxic baits of restricted use. Here we evaluate a push-pull strategy (i.e., the simultaneous use of attractant and repellent stimuli in order to divert pests) to manage LCAs Acromyrmex spp. in young willow plantations in the area of Delta of the Parana River, Argentina, a wetland ecosystem. First, we surveyed ants' selection of farmland vegetation during one year. Then, we estimated ants' preferences between the willow Salix babylonica and a subsample of plant species from farmland vegetation under laboratory conditions. Finally, we designed and performed a fully crossed experimental field assay to evaluate a push-pull strategy by using farmland vegetation as pull stimulus. RESULTS: We surveyed 39 plant species in the area, 19 of which had been foraged by LCAs along the year. Plants were selected by species, not by abundance. In the lab, ants showed similar preference for the cultivated willow and the subsample of plant species. Push-pull was the only treatment that maintained willow remaining vegetation above 60-80% at the end of the growing season. CONCLUSIONS: For the first time the push-pull strategy was evaluated in social insects. We demonstrated that it can be successfully used to manage LCAs in young willow plantations. Our strategy generates biodiversity, which can improve the ecosystem functioning, and it can be easily implemented by producers since its design is based on regular willow plantations.

Comparison of radial growth rate of the mutualistic fungus of Atta sexdens rubropilosa forel in two culture media.

In vitro culture of the mutualistic fungus of leaf-cutting ants is troublesome due to its low growth rate, which leads to storage problems and contaminants accumulation. This paper aims at comparing the radial growth rate of the mutualistic fungus of Atta sexdens rubropilosa Forel in two different culture media (Pagnocca B and MEA LP). Although total MEA LP radial growth was greater all along the bioassay, no significant difference was detected between growth efficiencies of the two media. Previous evidences of low growth rate for this fungus were confirmed. Since these data cannot point greater efficiency of one culture medium over the other, MEA LP medium is indicated for in vitro studies with this mutualistic fungus due its simpler composition and translucent color, making the analysis easier.

Nest Architecture and Colony Growth of Atta bisphaerica Grass-Cutting Ants.

Atta grass-cutting ants (Formicidae: Myrmicinae: Attini) are found in the Cerrado biome and build giant nests with hundreds or thousands of large chambers. We assessed for Atta bisphaerica grass-cutting ants whether the total volume of fungus chambers and other nest parameters grow at close or similar proportions to worker numbers in the colony. Data on fungus garden biomass, population, external area, number of entrance holes, number of fungus chambers, total fungus chambers volume, and nest depth were obtained during colony growth/nest development. Our results reveal that the fungus garden biomass, external area, and total fungus chamber volume increased at rates similar to the increase in the number of workers. The total volume of chambers, and external area increased at a similar proportion to the increase in number of workers, probably due to the fungus garden allocation needs of the colony. The number of fungus chambers, number of entrance holes, and nest depth increased less than 4-fold for every 10-fold increase in the worker number. In addition, the height of the fungus chambers increased as the width increased, a pattern not observed for tunnel height and length, and the chamber volume increased according to worker number. Thus, this study demonstrates for A. bisphaerica that nest development in terms of chamber volume is similar to the increase in number of colony workers, and this contributes to a better understanding of Atta nest architecture.

Effects of Entomopathogenic Fungi on Individuals as Well as Groups of Workers and Immatures of Atta sexdens rubropilosa Leaf-Cutting Ants.

In 2009, sulfluramid, the main ingredient in toxic baits for leaf-cutting ant control, was included in Annex B of the Stockholm Convention on Persistent Organic Pollutants. This resulted in interest in the use of entomopathogenic fungi such as Beauveria bassiana and Trichoderma harzianum for leaf-cutting ant control. The efficiency of these fungi in controlling these insects and the way that ants react individually or in group to the biological risks posed by these fungi is poorly understood. For this reason, we assessed the effects of B. bassiana and T. harzianum on Atta sexdens rubropilosa larvae, pupae and workers. Moreover, we investigated whether the number of contaminated individuals within a group has an influence in controlling the spread of fungi among workers. We found that the fungus T. harzianum showed high pathogenicity against A. sexdens rubropilosa larvae and pupae, leading to faster mortality and a survival rates. On the other hand, the fungus B. bassiana was responsible for causing faster worker mortality and lower survival rates. In addition, we observed that an increase in individuals contaminated with B. bassiana or T. harzianum in the group decreases its survival rate. The results support the hypothesis that entomopathogenic fungi are efficient in controlling leaf-cutting ants when contaminated workers are allocated to groups of healthy workers.

Selection and spatial arrangement of building materials during the construction of nest turrets by grass-cutting ants.

Ants build complex nest structures by reacting to simple, local stimuli. While underground nests result from the space generated by digging, some leaf- and grass-cutting ants also construct conspicuous aboveground turrets around nest openings. We investigated whether the selection of specific building materials occurs during turret construction in Acromyrmex fracticornis grass-cutting ants, and asked whether single building decisions at the beginning can modify the final turret architecture. To quantify workers' material selection, the original nest turret was removed and a choice between two artificial building materials, thin and thick sticks, was offered for rebuilding. Workers preferred thick sticks at the very beginning of turret construction, showed varying preferences thereafter, and changed to prefer thin sticks for the upper, final part of the turret, indicating that they selected different building materials over time to create a stable structure. The impact of a single building choice on turret architecture was evaluated by placing artificial beams that divided a colony's nest entrance at the beginning of turret rebuilding. Splitting the nest entrance led to the self-organized construction of turrets with branched galleries ending in multiple openings, showing that the spatial location of a single building material can strongly influence turret morphology.

Nest Turrets of Acromyrmex Grass-Cutting Ants: Micromorphology Reveals Building Techniques and Construction Dynamics.

Acromyrmex fracticornis grass-cutting ants construct conspicuous chimney-shaped nest turrets made of intermeshed grass fragments. We asked whether turrets are constructed by merely piling up nearby materials around the entrance, or whether ants incorporate different materials as the turret develops. By removing the original nest turrets and following their rebuilding process over three consecutive days, age-dependent changes in wall morphology and inner lining fabrics were characterized. Micromorphological descriptions based on thin sections of turret walls revealed the building behaviors involved. Ants started by collecting nearby twigs and dry grass fragments that are piled up around the nest entrance. Several large fragments held the structure like beams. As a net-like structure grew, soil pellets were placed in between the intermeshed plant fragments from the turret base to the top, reinforcing the structure. Concomitantly, the turret inner wall was lined with soil pellets, starting from the base. Therefore, the consolidation of the turret occurred both over time and from its base upwards. It is argued that nest turrets do not simply arise by the arbitrary deposition of nearby materials, and that workers selectively incorporate large materials at the beginning, and respond to the developing structure by reinforcing the intermeshed plant fragments over time.

Morphology of ovary and spermathecae of the parasitoid Eibesfeldtphora tonhascai Brown (Diptera: Phoridae).

Eibesfeldtphora tonhascai (Diptera: Phoridae) is a parasitoid of leaf-cutting ants and a potential biological control agent against these insect pests. This study describes the morphology of the ovary and spermatheca of E. tonhascai. The female reproductive tract of this parasitoid has a pair of meroistic polytrophic ovaries, two lateral oviducts that open into a common oviduct, an elongated accessory gland, and two spermathecae. Young oocytes are small and spherical, and their size increases as yolk is stored in the cytoplasm. This process is followed by chorion production by follicular cells. Mature oocytes are elliptical or torpedo-shaped. The reservoir wall of the spermatheca has type III glandular cells with cytoplasm rich in free ribosomes, rough endoplasmic reticulum, and secretory vesicles. The apical surface of these cells has microvilli associated with mitochondria. The reservoir lumen is lined by a cuticle and filled with spermatozoa. This is the first report of the ovary and spermatheca morphology of E. tonhascai and contributes to the comprehension of the reproductive biology of this parasitoid of leaf-cutting ants.

Increase in scout trips due to forager removal in Atta sexdens (Hymenoptera, Formicidae) (Forel, 1908).

Social information exchange through physical contacts and chemical trail deposition forms the basis of food recruitment in leaf-cutting ants. The scout initiates the process and passes the information to nestmates that recruit more foragers, thus amplifying the stimulus and ensuring the success of foraging. An interruption of the contact between workers and a reduction in trail laying can diminish the effectiveness of mass recruitment and alter scouting activity and forager flow. This study verified an increase in scout trips as a consequence of inbound workers (with or without a plant load) removal during Atta sexdens foraging, sustaining the outbound flow of foragers, and consequently foraging activity, either through direct contact or chemical trail deposition. Data indicate as one of the roles of unladen workers along the foraging trail must be to stimulate other workers to go out and so speed up the recruitment process The remarkable ability to organize themselves without central control is a major strength of social insects and the increase in scouting activity observed here is an example of this behavioral flexibility in leaf-cutting ants. Although foraging performance is enhanced through communication between workers, the simple adjustment in scouting activity can maintain the outbound flow of foragers which is an essential activity of the colony.

Vegetation Changes in a Native Forest Produced by Atta vollenweideri Forel 1893 (Hymenoptera: Formicidae) Nests.

Herbivory is an important factor to generate spatial mosaics with variations in a plant community composition and organization. The objective of this work was to determine the impact of Atta vollenweideri Forel 1893 nests on herbaceous and shrub vegetation in a degraded native forest of the Espinal ecoregion. The study was carried out in the Protected Area and Multiple Use Nature Reserve called Estancia "El Carayá" (Entre Ríos, Argentina). Ten A. vollenweideri nests were selected by simple random sampling through internal roads, and two transects were drawn from the center of the nest (0 m) up to 60 m away in opposite directions. The line intercept method was used to quantify the percentage of vegetation cover of herbaceous and shrub species, while the floristic composition was estimated by the Canfield method. Afterwards, a nonparametric test between positions and a conglomerate analysis to evaluated distance were applied. Grass species, legumes, and sedges fell in the adjacent areas to nests, highlighting the bare soil at the crest and base of the nests. Fifteen plant species were identified, and two families correspond to monocotyledonous and dicotyledonous species. In conclusion, the nests of A. vollenweideri affect the community of herbaceous and shrub vegetation of the studied degraded native forest of the Espinal ecoregion since these ants perform a high selection of herbaceous species considered as pioneers of plant successions.

Nest refuse of Acromyrmex balzani (Hymenoptera: Formicidae) increases the plant vigor in Turnera subulata (Turneraceae) / O substrato de descarte de Acromyrmex balzani (Hymenoptera: Formicidae) aumenta o vigor da planta Turnera subulata (Turneraceae)

Abstract 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.

Resumo Alguns estudos relatam o efeito positivo de resíduos orgânicos de formigueiros nas propriedades do solo e na estrutura da comunidade de plantas adjacentes em experimentos de campo, mas há uma lacuna sobre o efeito em espécies individuais. O objetivo do presente estudo foi comparar o desenvolvimento de Turnera subulata Smith, uma espécie ornamental, na presença do substrato de descarte (SD) de formigueiros produzido pela formiga cortadeira Acromyrmex balzani (Emery, 1890) por meio de experimento em casa de vegetação. O experimento foi conduzido com dois tratamentos: solo controle e solo com 25% de SD. As plantas foram mantidas em vasos de 1L por 90 dias. Foram avaliados os parâmetros: altura da planta, diâmetro do caule, comprimento da raiz, número de folhas, massa seca da raiz, biomassa seca e fresca da parte aérea. Além disso, o conteúdo relativo de clorofila e os nutrientes foliares foram usados como parâmetros fisiológicos. Como resultado, as plantas que cresceram no solo com SD apresentaram valores significativamente maiores de todos os parâmetros avaliados quando comparadas ao tratamento controle (p < 0.001). Concluímos que esse biofertilizante contribuiu para a produção de plantas mais vigorosas, podendo atuar na dinâmica local de nutrientes nos ecossistemas onde A. balzani ocorre. Além disso, por ser relativamente abundante e fácil de coletar, o SD de A. balzani tem potencial para ser utilizado como biofertilizante na produção de plantas de ciclo de vida curta.