Ant Assemblages Research Articles

Functional and genetic diversity changes through time in a cloud forest ant assemblage

AbstractClimate change in the Neotropics is causing upslope range shifts. We used arrays of ant species collected in a cloud forest at 1,500 m in Área de Conservación Guanacaste (ACG), northwestern Costa Rica, collected in two time periods (1998–2000 and 2008–2011) to measure changes in species richness and diversity over a decade. Using metrics of community structure, we found that the species assemblage in the collections from the 1990s was significantly phylogenetically clustered and functionally less diverse as compared to collections from the early 2000s. At both time points, the assemblages were significantly phylogenetically clustered and while the difference in functional diversity between the time points was not significant, the ant assemblage has become lighter in color (on average) over time. When individual species are considered, the overall pattern of replacement is consistent with the cloud forest ant assemblage being colonized by arrivals from lower elevation forests. The invertebrate communities on cloud forested mountain tops are especially vulnerable to a changing climate as there are two factors working together; no higher terrain to which they can move and the invasion of more and more taxa from lower downslope. This vulnerability is already measurable.

Monitoring ant assemblages of oak wood-pastures. A case study from Eastern Europe

Habitats associated with oak may host high insect diversity, especially saproxylic species, due to the availability of specific microhabitats such as dead wood, rotten logs and woody debris. Among the insects occurring in such habitats, ants play a major role in forest ecosystems as generalist predators. Ant assemblages were investigated using different types of traps. Sampling was carried out in 2015 and 2016, in a wood-pasture located in eastern Transylvania (Romania). Differences between species richness and composition in different types of oak habitats were tested: solitary oak in pasture, oak stand, and the ecotone separating them. Altogether a total of 17 ant species were identified. The three habitats shared an important proportion of the species, yet significant differences between the three habitats in terms of community composition were found. The highest diversity of ant species was recorded in the oak stands, followed by the solitary trees, though differences were not significant. Oak associated habitats can sustain high diversity of ant species and moreover thermophilic specialists thrive. Finally, the abundance of Lasius brunneus is highlighted, the species being proposed as a wood-pasture specialist.

Public Engagement Provides First Insights on Po Plain Ant Communities and Reveals the Ubiquity of the Cryptic Species Tetramorium immigrans (Hymenoptera, Formicidae).

Simple SummaryPublic involvement in biodiversity research in the form of Citizen Science is a powerful tool to improve our understanding of the natural world, and it is especially suitable for the study of heavily populated environments. Ants’ ubiquity and diversity, their role as ecological bioindicators, and the fact that most species can easily be sampled makes them ideal candidates for this kind of studies. In the framework of the international School of Ants citizen science project, we joined the “BioBlitz Lombardia” in which citizens are invited to collect biodiversity data on several parks from Lombardy (Po Plain, Italy). As a result, we recorded 30 ant species and obtained a first characterization of the region’s ant assemblages. We studied their patterns of variation in relation with the ecological difference between the studies sites, which ranged from urban to subalpine areas. In addition, we detected the presence of a cryptic species (Tetramorium immigrans) whose distribution and identity were only recently clarified. It likely represents an under-recorded introduced species in the region. Advantages and critical aspects of using CS methodology for the study of biodiversity are discussed in light of our experience.Ants are considered a useful model for biodiversity monitoring and several of their characteristics make them promising for citizen science (CS) projects. Involving a wide range of public figures into collecting valuable data on the effect of human impact on ant biodiversity, the School of Ants (SoA) project represents one of the very few attempts to explore the potential of these insects in CS. Through the collaboration with the “BioBlitz Lombardia” project, we tested the SoA protocol on 12 Northern Italy parks, ranging from urban green to subalpine protected sites. As a result, we obtained some of the very first quantitative data characterizing the ants of this region, recording 30 species and highlighting some interesting ecological patterns. These data revealed the ubiquitous presence of the recently taxonomically defined cryptic species Tetramorium immigrans, which appears to be probably introduced in the region. We also discuss advantages and criticisms encountered applying the SoA protocol, originally intended for schools, to new categories of volunteers, from BioBlitz participants to park operators, suggesting best practices based on our experience.

Species assemblages and diversity of litter ants (Hymenoptera: Formicidae) along an altitudinal gradient across two vegetation types in Hanthana Forest in Kandy, Sri Lanka

Ants account for 30% of terrestrial biomass and are highly sensitive to environmental variables. Present study investigated the effect of altitude (400-900 m) and vegetation type (mixed woodland and pine plantations in Hanthana forest) on litter ants. Two transects were laid at each of the five 100 m elevation levels. For hand-picking and litter extraction of ants, 1 m2 plots were laid at every 20 m intervals along the two sides of each transect. A total of 8689 ants in 9 subfamilies 32 genera and 56 morpho-species were collected. Subfamily Myrmicinae had the highest number of genera (15) and species (26 spp.). Genus Pheidole was represented by the highest number of species (5 spp.) while Solenopsis sp. 1 was the most widely distributed and abundant (6420 individuals) species. The mixed woodlands, regardless of elevation, were richer in ant species than pine plantations. Permutation test (ρ=0.30, P=3%) revealed a weak relationship between ant assemblages and environmental variables. Findings indicated that the difference in litter ant species assemblages and diversity in Hanthana Forest is possibly due to differences in vegetation types rather than altitude.

Effect of dominant parabioticAnt‐Gardenants on the understory and ground‐dwelling ant assemblage in the Amazon rainforest

ABSTRACTCamponotus femoratus(Fabricius, 1804) andCrematogaster leviorLongino, 2003 are Amazonian ants that jointly live in a parabiotic association in arboreal nests that are known as Ant‐Gardens (AGs). Both species are locally abundant and aggressive, which forage on the ground and on the vegetation.We evaluated, with extensive sampling, the effect ofCa. femoratusandCr. levioron the ground and the understory dwelling of ant assemblages in the southern Amazon using pit‐fall traps, beating‐tray sampling, and baiting.We found thatCa. femoratusandCr. leviorcaused a decrease in both the abundance and the species richness of the ground ants, changing the assemblage composition, and causing a nestedness pattern. Yet, AG ants seem to not promote significant changes in the assemblage of ants in the understory, with the exception of canopy openness and region, indicating a spatial resource partitioning.Our results showed that the frequency of these parabiotic AG ant pairs can modify the regional assemblage of the ground‐dwelling ants. As ants are ecological engineers and interact with numerous organisms in tropical habitats, the effect of the parabiotic ants can be extended to other organisms that inhabit the gaps along succession.

Distance–decay patterns differ between canopy and ground ant assemblages in a tropical rainforest

AbstractBoth decreases in compositional similarity with increasing geographic distances between sites (i.e. distance–decay relationship) and vertical stratification of species composition are key issues in ecology. However, the intersection between these two trends has scarcely been investigated. Here we use identical sampling methods in the canopy and at ground level in a tropical rainforest remnant on the coast of the Gulf of Mexico to evaluate, for the first time, a distance–decay relationship within vertical strata in insect assemblages. We found that the ant assemblage was vertically stratified; ant species richness was higher at ground level than in the canopy, and the species composition differed between the two vertical strata. Moreover, we observed that β-diversity increased with geographic distance at ground level, but not in the canopy strata. However, contrary to our prediction, there was less species turnover (lower β-diversity) between vertical strata than between trees. These findings may reflect differences in the dispersal capacity and nest habit of ants from each vertical stratum, and also habitat heterogeneity on the horizontal scale, e.g. the species of sampled trees. Our results illustrate the importance of sampling more than one vertical stratum to understand the spatial distribution patterns of biological diversity in tropical rainforests.

Ants (Hymenoptera: Formicidae) of Rockwood Park, New Brunswick: An Assessment of Species Richness and Habitat

Based on samples from 5 discrete habitat types (Thuja occidentalis [Eastern White Cedar] stand, Eastern White Cedar fen, old field, Acadian Mixed Forest, peat bog) we report the first assessment of ant species richness for a specific locale in New Brunswick. We identified 30 ant species across 3 subfamilies and 9 genera as present in Rockwood Park, a heavily wooded, 890-ha municipal park within the city of Saint John. All ant species encountered were native. Given our estimate of a total of 85–96 ant species present in New Brunswick (based on New Brunswick Museum records and studies of ants in northeastern North America), we found ∼35% in Rockwood Park. Major differences were evident in the assemblages of ant species among the 5 habitat types examined. While each habitat included generalist ant species shared across habitat types, ant species unique to each habitat were also present. Of the habitats studied in Rockwood Park, peat bogs and Eastern White Cedar stands supported the ant assemblages that were the most diverse, including several species that have rarely been encountered in North America. Both peat bogs and Eastern White Cedar stands in New Brunswick are widely exploited for commercial purposes. Our results suggest that environmental assessments of such habitats may need to consider the presence of rare, specialist ant species.

How estimated ant diversity is biased by the sampling method? A case study of Crete: a Mediterranean biodiversity hotspot

Defining biodiversity conservation goals requires representative and reliable data. However, data collected with the use of different methods can lead to divergent conclusions. Regardless of the high level of biodiversity of Mediterranean habitats, very little attention was paid in developing methods allowing rapid and scalable estimation of their richness. This study aims to recognize and explain the differences in performance of two methods: pitfall traps (PTM) and a complementary method based on hand collecting (HCM), in surveys of ants in the Mediterranean. We compared the results of applying both methods in three habitats in relation to α-, β-, and γ-diversity, and functional traits of species, i.e. Webber’s length (WL), nesting preferences, and evolutionary origin. Mean species number per HCM was significantly higher than per PTM sample. Spatial species turnover of HCM samples was higher than in PTM ones. However, assemblage dispersion did not differ. HCM detected a higher number of species and genera. WL differed significantly between HCM and PTM, and HCM recorded significantly more species in individual nesting guilds, regardless of considered habitat. HCM detected higher diversity of functional characteristic combinations of species. PTM detected fewer species with slightly larger body size and is useful in recognizing spatial species diversity patterns. HCM detected a higher number of species and produced more comprehensive results in identifying the functional diversity of ant assemblages. In conclusion, an integrated approach, described here as HCM, appears to be more suitable for heterogeneous Mediterranean habitats, especially when a survey aims at α-, β-, and γ-diversity assessments.

Habitat disturbance modifies dominance, coexistence, and competitive interactions in tropical ant communities

1. Interspecific competition is a major structuring principle in ecological communities. Despite their prevalence, the outcome of competitive interactions is hard to predict, highly context‐dependent, and multiple factors can modulate such interactions.2. We tested predictions concerning how competitive interactions are modified by anthropogenic habitat disturbance in ground‐foraging ant assemblages inhabiting fragmented Inter‐Andean tropical dry forests in southwestern Colombia, and investigated ant assemblages recruiting to baits in 10 forest fragments exposed to varying level of human disturbance.3. Specifically, we evaluated how different components of competitive interactions (patterns of species co‐occurrence, resource partitioning, numerical dominance, and interspecific trade‐offs between discovery and dominance competition) varied with level of habitat disturbance in a human‐dominated ecosystem.4. Multiple lines of evidence suggest that the role of competitive interactions in structuring ground‐foraging ant communities at baits varied with respect to habitat disturbance. As disturbance increased, community structure was more likely to exhibit random co‐occurrence patterns, higher levels of monopolization of food resources by dominant ants, and disproportionate dominance of a single species, the little fire ant (Wasmannia auropunctata). At a regional scale, we found evidence for a trade‐off between dominance and discovery abilities of the 15 most common species at baits.5. Together, these results suggest that human disturbance modifies the outcome of competitive interactions in ground‐foraging ant assemblages and may promote dominant species that reduce diversity and coexistence in tropical ecosystems.

Is ant fauna a good bio-indicator of coastal dune ecosystems in North-East of Algeria?

Summary Ants are good bioindicators of habitats and of their quality in many ecosystems. However, their interest in the case of coastal dunes remains to be determined because the environments are instable or subject to strong constraints. The degree of preservation of the Jijel coastal dune system is variable and presents an environmental gradient (sea-land) of five habitats defined by vegetation: upper beach, embryonic, mobile, steady and wooded dune. The aim of the study is to determine if (i) these five habitats have characteristic ant assemblages; (ii) if the variation in ant richness along the gradient shows a continuous increase reflecting very strong disturbances or shows a peak reached before the most stable habitat, therefore at the level of a medium disturbance; and (iii) how environmental factors play on the different ant species. Seventeen stations spread over four sites were selected for the study of myrmecofauna by barber trap, bait, and manual collection, and of vegetation by the quadrats method. Overall, the ant richness profile along the gradient is consistent with an increase towards the most stable environments. The composition of ant communities varies according to habitats and is correlated with the composition of plant communities. The effects on ants of environmental factors deduced from the structure of the vegetation are analysed at the community and species levels. Along the habitat gradient, there is a quadratic relationship between the barycentre and the range of habitat of the species. Despite the small number of species, this group of insects gives interesting results in terms of sensitivity to disturbances.

Indirect effect of the invasive exotic fungus Ophiostoma novo-ulmi (Dutch elm disease) on ants

Biotic invasions have become major agents of human-driven global change. One of the most virulent species for trees is the Dutch elm disease (DED) (Ophiostoma novo-ulmi). However, few are the studies that investigate its indirect impact on the fauna. In a well-preserved area of Sierra Morena (Cordoba, Spain), we report the repercussion of tree cover loss due to DED on ants (Hymenoptera, Formicidae), a bioindicator group. For 10 years, before and after DED, we studied ant assemblages in different zones with a gradient of invasion. In the most affected ones, indicator species of well-preserved habitats decreased, while generalist and opportunist species increased. The forest management practices undertaken after the disease had an even more devastating effect, and thus the logging and removal of dead trees in the Bejarano stream caused that its ant assemblage became more similar to the one of a nearby meadow than to the ones of close riparian areas.

Distribution of epigeic and hypogeic ants (Hymenoptera: Formicidae) in ombrophilous forests in the Brazilian Amazon

In the Amazon basin, ants are often associated with environmental or edaphic factors. However, these associations may vary between the epigeic and hypogeic strata. Here, we investigated differences in richness and composition of epigeic and hypogeic ant assemblages along an environmental gradient in the Brazilian Amazon. The four studied sites cover different topographic and soil characteristics. We sampled 25 plots of 250 m2 using 10 samples of epigeic pitfalls and 10 samples of hypogeic pitfalls installed at two depths (10 and 30 cm). The pitfalls remained in the fi eld for 48 hours. In the same plots, soil clay content and terrain altitude were also measured. We collected 219 species or morphospecies, of which 14 were exclusively hypogeics. We found higher local richness in the epigeic compared to hypogeic assemblages. We also found an interaction between clay content and strata for ant species composition. Overall, the species turnover was related to clay content, but the eff ect depended on the strata, with hypogeic fauna being more heterogeneous, compared with epigeic fauna. Despite the relationship between clay content and ant´s assemblage’s composition, we did not find strong environment predictors for both strata, which suggests that other factors may structure ant assemblages in these sites. This reinforces the need for studies to defi ne which environmental gradiente determines the distribution of Amazonian epigeic and hypogeic ants.

Fatores associados à ocorrência de formigas em escolas urbanas

As escolas consistem em ambientes que oferecem condições favoráveis para acolonização por formigas. Este estudo objetiva identificar os fatores associadosà ocorrência de formigas em quatro ambientes escolares localizados em áreasurbanas. A amostragem foi realizada em cozinhas, almoxarifados, refeitóriose áreas externas em doze escolas de quatro municípios da região oeste doestado de Santa Catarina, Brasil. Foram utilizadas iscas e coleta manual naamostragem. Variáveis ambientais foram obtidas em cada local. A similaridadeentre as amostras quanto à composição e a abundância de formigas foiavaliada por meio de uma escala multidimensional não métrica (non-metricmultidimensional scaling — NMDS). Foram amostrados 1.478 exemplaresde formigas e identificados 45 espécies. Foi observada uma similaridade de60% na abundância e composição das assembleias de formigas entre escolasde pequenas cidades (Caxambu do Sul, Guatambú e Palmitos). A presençade áreas externas com vegetação e áreas verdes no entorno, o tamanho daárea construída e a frequência de eventos de controle de insetos contribuempara a abundância desses insetos nos ambientes escolares. As formigas nãosão reconhecidas como potenciais vetores de micro-organismos patogênicosnas escolas e são negligenciadas se considerada a periodicidade do controlede insetos.

Omnivorous ants are less carnivorous and more protein-limited in exotic plantations.

Diets of species are crucial in determining how they influence food webs and community structures, and how their populations are regulated by different bottom-up processes. Omnivores are able to adjust their diet flexibly according to environmental conditions, such that their impacts on food webs and communities, and the macronutrients constraining their population, can be plastic. In particular, omnivore diets are known to be influenced by prey availability, which exhibits high spatial and temporal variation. To examine the plasticity of diet and macronutrient limitation in omnivores, we compared trophic positions, macronutrient preferences and food exploitation rates of omnivorous ants in invertebrate-rich (secondary forests) and invertebrate-poor (Lophostemon confertus plantations) habitats. We hypothesized that omnivorous ants would have lower trophic positions, enhanced protein limitation and reduced food exploitation rates in L. confertus plantations relative to secondary forests. We performed cafeteria experiments to examine changes in macronutrient limitation and food exploitation rates. We also sampled ants and conducted stable isotope analyses to investigate dietary shifts between these habitats. We found that conspecific ants were less carnivorous and had higher preferences for protein-rich food in L. confertus plantations compared to secondary forests. However, ant assemblages did not exhibit increased preferences for protein-rich food in L. confertus plantations. At the species-level, food exploitation rates varied idiosyncratically between habitats. At the assemblage-level, food exploitation rates were reduced in L. confertus plantations. Our results reveal that plantation establishments alter the diet and foraging behaviour of omnivorous ants. Such changes suggest that omnivorous ants in plantations will have reduced top-down impacts on prey communities but also see an increased importance of protein as a bottom-up force in constraining omnivore population sizes.

Enemy-free space and the distribution of ants, springtails and termites in the soil of one tropical rainforest

The soil fauna of tropical rainforests is difficult to study because of its extreme species richness and taxonomic impediment. Studies of multi-taxon assemblages in the soil of tropical rainforests are relatively rare and studies of interspecific interactions, such as predation, even rarer. Here we attempt to infer prey predator interactions and enemy-free space from the faunal composition of 100 litter/soil samples obtained from Barro Colorado Island in Panama during the dry and wet seasons. We focused on assemblages of ants (assigned to categories of non-predators, potential and confirmed predators), springtails and termites, which were characterized by their Barcode Index Numbers. Overall in 0.2 m3 of soil/litter we collected 2129 ants, 5592 springtails and 260 termites, which represented 80, 104 and 15 species, respectively. The faunal composition of confirmed ant predators was spatially coincident with that of Collembola. However, despite considerable seasonal shifts in the rank abundance of Collembola species, seasonal shifts of confirmed ant predators were low, resulting in a poor match of seasonal shifts between ants and their prey items. No location could be considered as being relatively free of ant enemies for springtails or termites, but the dry season supported higher prey-predator ratios than the wet season. We inferred only 4 possible prey-predator interactions, out of 7616 potential interactions in the study system. The relative dispersion of confirmed ant predators, which only weakly influenced springtail and termite assemblages, suggests low specificity in ant-prey interactions. This confirms that “brown food webs” may be structured by bottom-up effects rather than by top-down effects.

Mechanisms of species coexistence and functional diversity of ant assemblages in forest and pasture habitats in southwestern Brazilian Amazon

In this study, we investigated the mechanisms behind species coexistence and the relationships between functional diversity and species richness in ant assemblages in both forest and pasture habitats in the southwestern Brazilian Amazon. We addressed the specific question: What is the primary mechanism for species coexistence in forest and pasture habitats? According to the identified mechanism in each habitat, we had the following alternative expectations: (i) niche partitioning – we expected to observe a linear positive relationship between functional diversity and species richness, indicating a complementary relationship; or (ii) niche filtering – a positive constant asymptotic relation between functional diversity and species richness, indicating a functional redundancy relationship. In total, we sampled 91 ant species, 82 species in a forest habitat and 16, in a pasture habitat. In the forest habitat we identified niche filtering as the structuring mechanism of the ant assemblage, but we were unable to identify a clear mechanism in the pasture habitat. Although the relationship between functional diversity and species richness was positive in both habitats, the relationship was weaker in the forest habitat, indicating a greater functional redundancy among the ant species in this habitat. Our results reinforce the divergence of species coexistence mechanisms and ant assemblage structures in both natural and human-modified habitats in the Southwestern Brazilian Amazon.

Cohabitation and niche overlap in the occupation of twigs by arthropods in the leaf litter of Brazilian Atlantic Forest

In tropical forests, twigs are the nesting resources most frequently occupied by ants in the leaf litter. During occupation, this resource may be shared among different arthropods, such as ants or non-ant arthropods, but the mechanisms that promote cohabitation in this context are poorly known. In this study, we examined if twig structure influences species cohabitation. Additionally, we calculated co-occurrence and niche overlap metrics for ant species and non-ant arthropods occupying the same twigs in the leaf litter. We collected 52 species of ants and 82 morphospecies of non-ant arthropods inside 575 twigs from fragments of Brazilian Atlantic Forest. Thirty-six ant species cohabited with non-ant arthropods, which were primarily Isopoda and Coleoptera. We observed that the twigs most commonly shared by ants and non-ant arthropods were wider, longer, and had a larger number of holes. We also found that cohabiting ants and non-ant arthropods had higher co-occurrence and niche overlap scores. We suggest that twig diameter is an important factor determining occupation by ants and other arthropods. When the twig is occupied by more than one ant species, our results show that niche overlap and cohabitation with non-ant arthropods are less common. We conclude that examining twig structure is important to better understand arthropod species cohabitation in leaf litter twigs and to study coexistence in ant assemblages that use twigs.

Termite mounds create heterogeneity in invertebrate communities across a savanna rainfall gradient

Termite mounds create nutrient hotspots that serve as key resource areas for savanna vegetation and mammalian herbivores. However, despite the key ecological roles performed by termite mounds, few studies have investigated their influence on invertebrate communities, and none have examined such effects across environmental gradients. We hypothesised that termite mounds would support greater numbers of invertebrates than the surrounding savanna matrix and that assemblages would differ in composition due to the enhanced nutritional quality of vegetation on mounds. We also predicted that the differences between on-mound and off-mound invertebrate diversity would be more pronounced in areas where the difference in nutritional value between mounds and the savanna matrix vegetation was most prominent. We tested these hypotheses in Kruger National Park, South Africa, by sampling ground- and grass-dwelling invertebrate herbivores, omnivores and detritivores on and at various distances away from termite mounds at three savanna sites of varying vegetation quality across a rainfall gradient. All invertebrate groups sampled responded to termite mounds, but mound influence varied across trophic groups (Orthoptera showed the clearest patterns), diversity measures (changes in abundance rather than species richness) and with mean annual rainfall (strongest effects at the highest rainfall site). Orthoptera were more abundant on mounds, particularly at the wettest site, and there was a positive relationship between mound size and Orthoptera species richness. Ant assemblage composition on mounds differed from that off mounds and beetle abundance was greater on mounds, possibly as a result of concentrated mammalian herbivore activity and faecal deposition on mounds. Our results suggest that termite mounds are not only important nutrient and foraging hotspots for vertebrates, but that they also introduce heterogeneity in invertebrate communities, especially in nutrient-poor savannas.

Structure of ant assemblages on street trees in urban Busan, Korea

AbstractStreet trees planted in urban areas are one of the smallest urban green habitats; however, their role as a valuable refuge for local biota is poorly understood. The aim of this study was to investigate urban street trees as habitat for ants. We found ants on 195 (92%) of 211 street trees studied. Seventeen ant species were collected from five tree species. The two most common ant species were Tetramorium tsushimae (on 112 trees) and Lasius japonicus (on 93 trees). The number of ant species found per tree ranged from 0 to 5, with a mean of 1.55 ± 0.92. The species composition differed significantly among tree species. Nine ant species (Camponotus vitiosus, Lasius alienus, L. japonicus, Nylanderia flavipes, Crematogaster matsumurai, Crematogaster vagula, Monomorium intrudens, T. tsushimae, and Brachyponera chinensis) nested on the trees, mostly in the root zone. Nests were found on 150 trees but were relatively rare on trees with diameter at breast height of less than 40 cm.

Accessing the subterranean ant fauna (Hymenoptera: Formicidae) in native and modified subtropical landscapes in the Neotropics

Abstract: Several studies have characterized and delimited subterranean ant assemblages. Soil extraction, one of the methods employed to access this fauna, employs the removal of monoliths. One of the most widely used methods for the extraction of soil sampling is called TSBF (developed by the Tropical Soil Biology and Fertility Programme). This method provides relevant data about the species associated with the soil. In the present study we characterized assemblages of subterranean ants using the TSBF method in different subtropical areas of the Neotropics. We considered two sampling designs in different localities. The first design resulted in 315 TSBF samples obtained from layers at distinct depths. The second design resulted in 270 TSBF samples and 270 epigaeic pitfall trap samples. This material was used to delimit the species that occur exclusively in the subterranean stratum (TSBF) and that are not found on the soil surface. A total of 281 species were recorded. Of these, 57 can be considered subterranean, based on their occurrence in subterranean strata. We also verified that the highest occurrence of ants was in the first 10 cm of soil depth. Due to the importance of using methods that efficiently extract the subterranean ant fauna in studies, we suggest the TSBF method should be used to sample ants or to associate this method with epigaeic pitfall traps to delimit strictly subterranean assemblages in specific community stratification studies.