Unforeseen plant phenotypic diversity in a dry and grazed world.

Earth harbours an extraordinary plant phenotypic diversity1 that is at risk from ongoing global changes2,3. However, it remains unknown how increasing aridity and livestock grazing pressure-two major drivers of global change4-6-shape the trait covariation that underlies plant phenotypic diversity1,7. Here we assessed how covariation among 20 chemical and morphological traits responds to aridity and grazing pressure within global drylands. Our analysis involved 133,769 trait measurements spanning 1,347 observations of 301 perennial plant species surveyed across 326 plots from 6 continents. Crossing an aridity threshold of approximately 0.7 (close to the transition between semi-arid and arid zones) led to an unexpected 88% increase in trait diversity. This threshold appeared in the presence of grazers, and moved toward lower aridity levels with increasing grazing pressure. Moreover, 57% of observed trait diversity occurred only in the most arid and grazed drylands, highlighting the phenotypic uniqueness of these extreme environments. Our work indicates that drylands act as a global reservoir of plant phenotypic diversity and challenge the pervasive view that harsh environmental conditions reduce plant trait diversity8-10. They also highlight that many alternative strategies may enable plants to cope with increases in environmental stress induced by climate change and land-use intensification.

Long-term strict ant-plant mutualism identity characterises growth rate and leaf shearing resistance of an Amazonian myrmecophyte.

Over 125 million years of ant-plant interactions have culminated in one of the most intriguing evolutionary outcomes in life history. The myrmecophyte Duroia hirsuta (Rubiaceae) is known for its mutualistic association with the ant Myrmelachista schumanni and several other species, mainly Azteca, in the north-western Amazon. While both ants provide indirect defences to plants, only M. schumanni nests in plant domatia and has the unique behaviour of clearing the surroundings of its host tree from heterospecific plants, potentially increasing resource availability to its host. Using a 12-year survey, we asked how the continuous presence of either only M. schumanni or only Azteca spp. benefits the growth and defence traits of host trees. We found that the continuous presence of M. schumanni improved relative growth rates and leaf shearing resistance of Duroia better than trees with Azteca. However, leaf herbivory, dry matter content, trichome density, and secondary metabolite production were the same in all trees. Survival depended directly on ant association (> 94% of trees died when ants were absent). This study extends our understanding of the long-term effects of strict ant-plant mutualism on host plant traits in the field and reinforces the use of D. hirsuta-M. schumanni as a model system suitable for eco-co-evolutionary research on plant-animal interactions.

Passive accumulation of alkaloids in non-toxic frogs challenges paradigms of the origins of acquired chemical defenses.

Understanding the origins of novel, complex phenotypes is a major goal in evolutionary biology. Poison frogs of the family Dendrobatidae have evolved the novel ability to acquire alkaloids from their diet for chemical defense at least three times. However, taxon sampling for alkaloids has been biased towards colorful species, without similar attention paid to inconspicuous ones that are often assumed to be undefended. As a result, our understanding of how chemical defense evolved in this group is incomplete. Here we provide new data showing that, in contrast to previous studies, species from each undefended poison frog clade have measurable yet low amounts of alkaloids. We confirm that undefended dendrobatids regularly consume mites and ants, which are known sources of alkaloids. Further, we confirm the presence of alkaloids in two putatively non-toxic frogs from other families. Our data suggest the existence of a phenotypic intermediate between toxin consumption and sequestration-passive accumulation-that differs from active sequestration in that it involves no derived forms of transport and storage mechanisms yet results in low levels of toxin accumulation. We discuss the concept of passive accumulation and its potential role in the origin of chemical defenses in poison frogs and other toxin-sequestering organisms.

Hotspots of biogeochemical activity linked to aridity and plant traits across global drylands.

Perennial plants create productive and biodiverse hotspots, known as fertile islands, beneath their canopies. These hotspots largely determine the structure and functioning of drylands worldwide. Despite their ubiquity, the factors controlling fertile islands under conditions of contrasting grazing by livestock, the most prevalent land use in drylands, remain virtually unknown. Here we evaluated the relative importance of grazing pressure and herbivore type, climate and plant functional traits on 24 soil physical and chemical attributes that represent proxies of key ecosystem services related to decomposition, soil fertility, and soil and water conservation. To do this, we conducted a standardized global survey of 288 plots at 88 sites in 25 countries worldwide. We show that aridity and plant traits are the major factors associated with the magnitude of plant effects on fertile islands in grazed drylands worldwide. Grazing pressure had little influence on the capacity of plants to support fertile islands. Taller and wider shrubs and grasses supported stronger island effects. Stable and functional soils tended to be linked to species-rich sites with taller plants. Together, our findings dispel the notion that grazing pressure or herbivore type are linked to the formation or intensification of fertile islands in drylands. Rather, our study suggests that changes in aridity, and processes that alter island identity and therefore plant traits, will have marked effects on how perennial plants support and maintain the functioning of drylands in a more arid and grazed world.

Nutrients or resin? - The relationship between resin and food foraging in stingless bees.

Stingless bees are important pollinators in tropical forests. Yet, we know little about their foraging behavior (e.g., their nutritional requirements or their floral sources visited for resource collection). Many stingless bees not only depend vitally on pollen and nectar for food but also on resin for nest building and/or defense. However, it is unclear whether the large effort devoted to collecting resin as a non-food resource by certain stingless bees affects their foraging behavior. Therefore, in this study, we analyzed differences in foraging patterns (i.e., foraging activity, proportion of collected resources, and specialization in plants visited) and resource nutritional composition (i.e., sucrose amount in nectar and amino acids in pollen) of seven different stingless bee species (eleven wild colonies) in north-western Ecuador with a particular focus on the role of resin collection. We found that species with a high resin intake tended to be more active than species with a low resin intake. The foragers per minute invested for pollen collection were similar across all species. Sucrose intake per minute differed between some species but was not affected by increased resin intake. Interestingly, high and low resin collectors partly differed in the plants visited for pollen collection. Pollen amino acid profiles largely, but not completely, overlapped between the two resin collection groups. Our findings show that the foraging patterns and plant choices of stingless bees may vary depending on their resin intake, highlighting the need for more research focusing on resin collection and use by stingless bees.

Las abejas sin aguijón son polinizadores importantes en los bosques tropicales. Sin embargo, sabemos poco acerca de su comportamiento de forrajeo (e.g., sus requisitos nutricionales o las fuentes florales visitadas para la recolección de recursos). Muchas abejas sin aguijón dependen vitalmente no solo de polen y de néctar como alimento, sino también de resinas para la construcción de su nido y/o defensa. Sin embargo, no está claro si el gran esfuerzo dedicado a la recolección de resina como recurso no alimentario de ciertas abejas sin aguijón afecta su comportamiento de forrajeo. Por lo tanto, en este estudio, analizamos las diferencias en los patrones de forrajeo (i.e., actividad de forrajeo, proporción de recursos recolectados y especialización en las plantas visitadas) y la composición nutricional de los recursos recolectados (i.e., cantidad de sacarosa en el néctar y de aminoácidos en el polen) de siete especies diferentes de abejas sin aguijón (once colonias silvestres) en el noroeste de Ecuador, con un enfoque particular en el rol de la recolección de resina. Encontramos que las especies con una recolección alta de resina tienden a ser más activas que las especies con una recolección baja de resina. La cantidad de forrajeadores por minuto dedicada a la recolección de polen fue similar en todas las especies. La ingesta de sacarosa por minuto difirió entre algunas especies, pero no se vio afectada por un aumento en la recolección de resina. Interesantemente, las abejas con una recolección alta y baja de resina difirieron parcialmente en las plantas que visitaron para la recolección de polen. Entre los dos grupos de recolección de resina también hubo diferencias con respecto al perfil de aminoácidos en el polen que recolectaron. El perfil de aminoácidos se sobrelapaba, pero no completamente, entre los dos grupos. Nuestros resultados muestran que los patrones de forrajeo y las elecciones de plantas de las abejas sin aguijón pueden variar según su consumo de resina, destacando la necesidad de hacer más investigaciones centradas en la recolección y el uso de resina por parte de las abejas sin aguijón.

Effects of leaf traits of tropical trees on the abundance and body mass of herbivorous arthropod communities.

In tropical forests, herbivorous arthropods remove between 7% up to 48% of leaf area, which has forced plants to evolve defense strategies. These strategies influence the palatability of leaves. Palatability, which reflects a syndrome of leaf traits, in turn influences both the abundance and the mean body mass not only of particular arthropod taxa but also of the total communities. In this study, we tested two hypotheses: (H1) The abundance of two important chewer guilds ('leaf chewers' and 'rostrum chewers'), dominant components of arthropod communities, is positively related to the palatability of host trees. (H2) Lower palatability leads to an increased mean body mass of chewers (Jarman-Bell principle). Arthropods were collected by fogging the canopies of 90 tropical trees representing 31 species in three plots at 1000 m and three at 2000 m a.s.l. Palatability was assessed by measuring several 'leaf traits' of each host tree and by conducting a feeding trial with the generalist herbivore Gryllus assimilis (Orthoptera, Gryllidae). Leaf traits provided partial support for H1, as abundance of leaf chewers but not of rostrum chewers was positively affected by the experimentally estimated palatability. There was no support for H2 as neither leaf traits nor experimentally estimated palatability affected the mean body mass of leaf chewers. The mean body mass of rostrum chewers was positively related to palatability. Thus, leaf traits and experimentally estimated palatability influenced the abundance and mean body mass of chewing arthropods on the community level. However, the data were not consistent with the Jarman-Bell principle. Overall, our results suggest that the palatability of leaves is not among the dominant factors influencing abundance and mean body mass of the community of chewing arthropod herbivores. If other factors, such as the microclimate, predation or further (a-)biotic interactions are more important has to be analyzed in refined studies.

Soundscapes and deep learning enable tracking biodiversity recovery in tropical forests.

Tropical forest recovery is fundamental to addressing the intertwined climate and biodiversity loss crises. While regenerating trees sequester carbon relatively quickly, the pace of biodiversity recovery remains contentious. Here, we use bioacoustics and metabarcoding to measure forest recovery post-agriculture in a global biodiversity hotspot in Ecuador. We show that the community composition, and not species richness, of vocalizing vertebrates identified by experts reflects the restoration gradient. Two automated measures - an acoustic index model and a bird community composition derived from an independently developed Convolutional Neural Network - correlated well with restoration (adj-R² = 0.62 and 0.69, respectively). Importantly, both measures reflected composition of non-vocalizing nocturnal insects identified via metabarcoding. We show that such automated monitoring tools, based on new technologies, can effectively monitor the success of forest recovery, using robust and reproducible data.

Catalog of the invertebrate type specimens hosted at the Pontificia Universidad Católica del Ecuador and Escuela Politécnica Nacional natural history collections.

This work updates the invertebrate type specimen catalog published by Donoso et al. (2009). The catalog is increased by 2281 type specimens (from 454 species or subspecies) to a total of 4180 type specimens (from 770 species or subspecies) hosted at the Pontificia Universidad Católica del Ecuador and Escuela Politécnica Nacional natural history collections. The new material adds 307 holotypes, 1910 paratypes, and 64 allotypes. It provides original information from four phyla (Arthropoda, Mollusca, Nemata, and Platyhelminthes), eight classes, 21 orders, 73 families, and 156 genera. This updated catalog includes a map showing the type localities in the country, a list of the 71 new type specimens (from 23 species or subspecies) from other countries hosted at both museums, corrections to the previous catalog published by Donoso et al. (2009), and label information from each new specimen.

Stratification and recovery time jointly shape ant functional reassembly in a neotropical forest.

Microhabitat differentiation of species communities such as vertical stratification in tropical forests contributes to species coexistence and thus biodiversity. However, little is known about how the extent of stratification changes during forest recovery and influences community reassembly. Environmental filtering determines community reassembly in time (succession) and in space (stratification), hence functional and phylogenetic composition of species communities are highly dynamic. It is poorly understood if and how these two concurrent filters-forest recovery and stratification-interact. In a tropical forest chronosequence in Ecuador spanning 34 years of natural recovery, we investigated the recovery trajectory of ant communities in three overlapping strata (ground, leaf litter, lower tree trunk) by quantifying 13 traits, as well as the functional and phylogenetic diversity of the ants. We expected that functional and phylogenetic diversity would increase with recovery time and that each ant community within each stratum would show a distinct functional reassembly. We predicted that traits related to ant diet would show divergent trajectories reflecting an increase in niche differentiation with recovery time. On the other hand, traits related to the abiotic environment were predicted to show convergent trajectories due to a more similar microclimate across strata with increasing recovery age. Most of the functional traits and the phylogenetic diversity of the ants were clearly stratified, confirming previous findings. However, neither functional nor phylogenetic diversity increased with recovery time. Community-weighted trait means had complex relationships to recovery time and the majority were shaped by a statistical interaction between recovery time and stratum, confirming our expectations. However, most trait trajectories converged among strata with increasing recovery time regardless of whether they were related to ant diet or environmental conditions. We confirm the hypothesized interaction among environmental filters during the functional reassembly in tropical forests. Communities in individual strata respond differently to recovery, and possible filter mechanisms likely arise from both abiotic (e.g. microclimate) and biotic (e.g. diet) conditions. Since vertical stratification is prevalent across animal and plant taxa, our results highlight the importance of stratum-specific analysis in dynamic ecosystems and may generalize beyond ants.

The global distribution of known and undiscovered ant biodiversity.

Invertebrates constitute the majority of animal species and are critical for ecosystem functioning and services. Nonetheless, global invertebrate biodiversity patterns and their congruences with vertebrates remain largely unknown. We resolve the first high-resolution (~20-km) global diversity map for a major invertebrate clade, ants, using biodiversity informatics, range modeling, and machine learning to synthesize existing knowledge and predict the distribution of undiscovered diversity. We find that ants and different vertebrate groups have distinct features in their patterns of richness and rarity, underscoring the need to consider a diversity of taxa in conservation. However, despite their phylogenetic and physiological divergence, ant distributions are not highly anomalous relative to variation among vertebrate clades. Furthermore, our models predict that rarity centers largely overlap (78%), suggesting that general forces shape endemism patterns across taxa. This raises confidence that conservation of areas important for small-ranged vertebrates will benefit invertebrates while providing a "treasure map" to guide future discovery.

Comparison of traditional and DNA metabarcoding samples for monitoring tropical soil arthropods (Formicidae, Collembola and Isoptera).

The soil fauna of the tropics remains one of the least known components of the biosphere. Long-term monitoring of this fauna is hampered by the lack of taxonomic expertise and funding. These obstacles may potentially be lifted with DNA metabarcoding. To validate this approach, we studied the ants, springtails and termites of 100 paired soil samples from Barro Colorado Island, Panama. The fauna was extracted with Berlese-Tullgren funnels and then either sorted with traditional taxonomy and known, individual DNA barcodes ("traditional samples") or processed with metabarcoding ("metabarcoding samples"). We detected 49 ant, 37 springtail and 34 termite species with 3.46 million reads of the COI gene, at a mean sequence length of 233 bp. Traditional identification yielded 80, 111 and 15 species of ants, springtails and termites, respectively; 98%, 37% and 100% of these species had a Barcode Index Number (BIN) allowing for direct comparison with metabarcoding. Ants were best surveyed through traditional methods, termites were better detected by metabarcoding, and springtails were equally well detected by both techniques. Species richness was underestimated, and faunal composition was different in metabarcoding samples, mostly because 37% of ant species were not detected. The prevalence of species in metabarcoding samples increased with their abundance in traditional samples, and seasonal shifts in species prevalence and faunal composition were similar between traditional and metabarcoding samples. Probable false positive and negative species records were reasonably low (13-18% of common species). We conclude that metabarcoding of samples extracted with Berlese-Tullgren funnels appear suitable for the long-term monitoring of termites and springtails in tropical rainforests. For ants, metabarcoding schemes should be complemented by additional samples of alates from Malaise or light traps.

Male ant reproductive investment in a seasonal wet tropical forest: Consequences of future climate change.

Tropical forests sustain many ant species whose mating events often involve conspicuous flying swarms of winged gynes and males. The success of these reproductive flights depends on environmental variables and determines the maintenance of local ant diversity. However, we lack a strong understanding of the role of environmental variables in shaping the phenology of these flights. Using a combination of community-level analyses and a time-series model on male abundance, we studied male ant phenology in a seasonally wet lowland rainforest in the Panama Canal. The male flights of 161 ant species, sampled with 10 Malaise traps during 58 consecutive weeks (from August 2014 to September 2015), varied widely in number (mean = 9.8 weeks, median = 4, range = 1 to 58). Those species abundant enough for analysis (n = 97) flew mainly towards the end of the dry season and at the start of the rainy season. While litterfall, rain, temperature, and air humidity explained community composition, the time-series model estimators elucidated more complex patterns of reproductive investment across the entire year. For example, male abundance increased in weeks when maximum daily temperature increased and in wet weeks during the dry season. On the contrary, male abundance decreased in periods when rain receded (e.g., at the start of the dry season), in periods when rain fell daily (e.g., right after the beginning of the wet season), or when there was an increase in the short-term rate of litterfall (e.g., at the end of the dry season). Together, these results suggest that the BCI ant community is adapted to the dry/wet transition as the best timing of reproductive investment. We hypothesize that current climate change scenarios for tropical regions with higher average temperature, but lower rainfall, may generate phenological mismatches between reproductive flights and the adequate conditions needed for a successful start of the colony.

Rapid ant community reassembly in a Neotropical forest: Recovery dynamics and land-use legacy.

Regrowing secondary forests dominate tropical regions today, and a mechanistic understanding of their recovery dynamics provides important insights for conservation. In particular, land-use legacy effects on the fauna have rarely been investigated. One of the most ecologically dominant and functionally important animal groups in tropical forests are the ants. Here, we investigated the recovery of ant communities in a forest-agricultural habitat mosaic in the Ecuadorian Chocó region. We used a replicated chronosequence of previously used cacao plantations and pastures with 1-34 years of regeneration time to study the recovery dynamics of species communities and functional diversity across the two land-use legacies. We compared two independent components of responses on these community properties: resistance, which is measured as the proportion of an initial property that remains following the disturbance; and resilience, which is the rate of recovery relative to its loss. We found that compositional and trait structure similarity to old-growth forest communities increased with regeneration age, whereas ant species richness remained always at a high level along the chronosequence. Land-use legacies influenced species composition, with former cacao plantations showing higher resemblance to old-growth forests than former pastures along the chronosequence. While resistance was low for species composition and high for species richness and traits, all community properties had similarly high resilience. In essence, our results show that ant communities of the Chocó recovery rapidly, with former cacao reaching predicted old-growth forest community levels after 21 years and pastures after 29 years. Recovery in this community was faster than reported from other ecosystems and was likely facilitated by the low-intensity farming in agricultural sites and their proximity to old-growth forest remnants. Our study indicates the great recovery potential for this otherwise highly threatened biodiversity hotspot.

Biodiversity and ecosystem functions depend on environmental conditions and resources rather than the geodiversity of a tropical biodiversity hotspot.

Biodiversity and ecosystem functions are highly threatened by global change. It has been proposed that geodiversity can be used as an easy-to-measure surrogate of biodiversity to guide conservation management. However, so far, there is mixed evidence to what extent geodiversity can predict biodiversity and ecosystem functions at the regional scale relevant for conservation planning. Here, we analyse how geodiversity computed as a compound index is suited to predict the diversity of four taxa and associated ecosystem functions in a tropical mountain hotspot of biodiversity and compare the results with the predictive power of environmental conditions and resources (climate, habitat, soil). We show that combinations of these environmental variables better explain species diversity and ecosystem functions than a geodiversity index and identified climate variables as more important predictors than habitat and soil variables, although the best predictors differ between taxa and functions. We conclude that a compound geodiversity index cannot be used as a single surrogate predictor for species diversity and ecosystem functions in tropical mountain rain forest ecosystems and is thus little suited to facilitate conservation management at the regional scale. Instead, both the selection and the combination of environmental variables are essential to guide conservation efforts to safeguard biodiversity and ecosystem functions.

Nutritional niches reveal fundamental domestication trade-offs in fungus-farming ants.

During crop domestication, human farmers traded greater productivity for higher crop vulnerability outside specialized cultivation conditions. We found a similar domestication trade-off across the major co-evolutionary transitions in the farming systems of attine ants. First, the fundamental nutritional niches of cultivars narrowed over ~60 million years of naturally selected domestication, and laboratory experiments showed that ant farmers representing subsequent domestication stages strictly regulate protein harvest relative to cultivar fundamental nutritional niches. Second, ants with different farming systems differed in their abilities to harvest the resources that best matched the nutritional needs of their fungal cultivars. This was assessed by quantifying realized nutritional niches from analyses of items collected from the mandibles of laden ant foragers in the field. Third, extensive field collections suggest that among-colony genetic diversity of cultivars in small-scale farms may offer population-wide resilience benefits that species with large-scale farming colonies achieve by more elaborate and demanding practices to cultivate less diverse crops. Our results underscore that naturally selected farming systems have the potential to shed light on nutritional trade-offs that shaped the course of culturally evolved human farming.

Platyhypnidium aquaticum as Bioindicator of Metal and Metalloid Contamination of River Water in a Neotropical Mountain City.

Water contamination is a major environmental problem in many cities of the world. Most water contamination results from industry and human activities that generate toxic substances (e.g., metals). Rheophilic and aquatic mosses are found in lotic ecosystems, and their morphological and physiological traits are responsive to ecological and pollution gradients. Here we hypothesized that the native rheophilic moss Platyhypnidium aquaticum (A. Jaeger) M. Fleisch exposed to polluted waters can bioaccumulate greater amounts of metals, and a metalloid, than P. aquaticum exposed to pollution-free water. To this aim, we tested the bioindicator capacity of the aquatic P. aquaticum for 15 metals (Cd, Pb, Zn, Fe, K, Ca, Na, Mn, V, Co, Ba, Cr, Al, Sr, and Mg) and one metalloid (As), in twelve river samples coming from three urban and one control zone along the Zamora river in the city of Loja. When compared to the control, our results showed that P. aquaticum in the Southern, Central, and Northern zones of the city bioaccumulated higher concentrations of Ba, Cd, Co, Fe, Mg, Mn, Na, Sr, Zn, and the metalloid As. On the other hand, concentrations of Al, Ca, Cr, Pb, and V in P. aquaticum tended to be lower in the control zone, but these differences were not significant. We suggest that the presence of these contaminants may be related to water pollution (e.g., residual discharges and a lack of treatment systems) along urban zones of the river. We report for the first time the utility of P. aquaticum as a model species for development of long-term biomonitoring programs of water contamination in South America. Passive biomonitoring with P. aquaticum can be a simple and low-cost method to obtain reliable data of the current state of water contamination with metals and metalloids in tropical regions.

Odontomachus davidsoni sp. nov. (Hymenoptera, Formicidae), a new conspicuous trap-jaw ant from Ecuador.

One of the largest species in its genus, Odontomachus davidsoni Hoenle, Lattke & Donoso, sp. nov. is described from workers and queens collected at lowland forests in the Chocó-Darién bioregion in coastal Ecuador. The workers are characterized by their uniform red coloration, their large size (16-18 mm body length), and their frontal head striation that reaches the occipital margin. DNA barcodes (COI) and high resolution 2D images of the type material are provided, as well as an updated key for the Neotropical species of Odontomachus. In addition, a three-dimensional digital model of the worker holotype and a paratype queen scanned with DISC3D based on photogrammetry is presented, for the first time in a species description. Findings of large and conspicuous new species are uncommon around the world and suggest that these Ecuadorian rainforests may conceal many more natural treasures that deserve conservation.

ResumenDescribimos una especie nueva, entre las más grandes conocidas del género Odontomachus. La nueva especie, Odontomachus davidsoni Hoenle, Lattke & Donoso, sp. nov., es descrita a partir de obreras y reinas recolectadas en bosques de tierras bajas en la bioregión Chocó-Darién de la costa del Ecuador. Las obreras se caracterizan por su coloración rojiza uniforme, su grande tamaño (largo del cuerpo 16­18 mm), y la estriación del frente cefálico que alcanza el margen occipital. Proveemos códigos de barras de DNA (COI) e imágenes 2D de alta resolución para el material tipo y así como una guía de identificación actualizada para las especies neotropicales del género Odontomachus. Por primera vez en una descripción de especies, se proveen imágenes 3D de un escáner fotogramétrico DISC3D. Los hallazgos de especies grandes y conspicuas son poco comunes alrededor del mundo y sugieren que estos bosques lluviosos ecuatorianos pueden contener muchos más tesoros naturales que merecen ser conservados.

Species-level predation network uncovers high prey specificity in a Neotropical army ant community.

Army ants are among the top arthropod predators and considered keystone species in tropical ecosystems. During daily mass raids with many thousand workers, army ants hunt live prey, likely exerting strong top-down control on prey species. Many tropical sites exhibit a high army ant species diversity (>20 species), suggesting that sympatric species partition the available prey niches. However, whether and to what extent this is achieved has not been intensively studied yet. We therefore conducted a large-scale diet survey of a community of surface-raiding army ants at La Selva Biological Station in Costa Rica. We systematically collected 3,262 prey items from eleven army ant species (genera Eciton, Nomamyrmex and Neivamyrmex). Prey items were classified as ant prey or non-ant prey. The prey nearly exclusively consisted of other ants (98%), and most booty was ant brood (87%). Using morphological characters and DNA barcoding, we identified a total of 1,103 ant prey specimens to the species level. One hundred twenty-nine ant species were detected among the army ant prey, representing about 30% of the known local ant diversity. Using weighted bipartite network analyses, we show that prey specialization in army ants is unexpectedly high and prey niche overlap very small. Besides food niche differentiation, we uncovered a spatiotemporal niche differentiation in army ant raid activity. We discuss competition-driven multidimensional niche differentiation and predator-prey arms races as possible mechanisms underlying prey specialization in army ants. By combining systematic prey sampling with species-level prey identification and network analyses, our integrative approach can guide future research by portraying how predator-prey interactions in complex communities can be reliably studied, even in cases where morphological prey identification is infeasible.