Molecular phylogeny of Vincetoxicum (Apocynaceae, Asclepiadoideae) from Thailand and integrative taxonomy corroborating a new cryptic species within Vincetoxicum kerrii.

An updated phylogeny of the genus Vincetoxicum s.l. based on DNA sequences of the nuclear internal transcribed spacer (ITS) region and three plastid markers is presented. In total, 21 accessions newly sequenced from Thailand were added to the dataset of the homologous sequences of 75 other Vincetoxicum taxa downloaded from GenBank. In our analysis, the relationships between the well-supported clades largely correspond to those revealed in previous studies. With some exceptions, the phylogenetic positions of the Thai taxa in relation to other conspecifics and congeners generally reflect the geographic distributions of taxa. Moreover, recent extensive sampling throughout Thailand and in-depth investigation have revealed V. kerrii, a slender twiner widespread from South China to Indo-China, to be a species complex. A combination of molecular, morphological, anatomical, ultrastructural and ecological evidence allowed us to reveal a new cryptic species hidden within V. kerrii, described here under the name V. simplex. A comprehensive description, illustrations, photographs, and comparison with the morphologically similar species are provided. Although V. simplex and V. kerrii s.s. resemble one another in various aspects of vegetative and reproductive structures, the latter is phylogenetically closely related to V. irrawadense, which is much less similar morphologically to both V. simplex and V. kerrii s.s. than the latter two are to each other. In addition to the new cryptic species recognized in the present study, a new record for Thailand, V. microstachys, is also reported.

A phylogenetic perspective on the association between ants (Hymenoptera: Formicidae) and black yeasts (Ascomycota: Chaetothyriales).

The frequency and the geographical extent of symbiotic associations between ants and fungi of the order Chaetothyriales have been highlighted only recently. Using a phylogenetic approach based on seven molecular markers, we showed that ant-associated Chaetothyriales are scattered through the phylogeny of this order. There was no clustering according to geographical origin or to the taxonomy of the ant host. However, strains tended to be clustered according to the type of association with ants: strains from ant-made carton and strains from plant cavities occupied by ants ('domatia') rarely clustered together. Defining molecular operational taxonomic units (MOTUs) with an internal transcribed spacer sequence similarity cut-off of 99% revealed that a single MOTU could be composed of strains collected from various ant species and from several continents. Some ant-associated MOTUs also contained strains isolated from habitats other than ant-associated structures. Altogether, our results suggest that the degree of specialization of the interactions between ants and their fungal partners is highly variable. A better knowledge of the ecology of these interactions and a more comprehensive sampling of the fungal order are needed to elucidate the evolutionary history of mutualistic symbioses between ants and Chaetothyriales.

Characterization of actinobacteria associated with three ant-plant mutualisms.

Ant-plant mutualisms are conspicuous and ecologically important components of tropical ecosystems that remain largely unexplored in terms of insect-associated microbial communities. Recent work has revealed that ants in some ant-plant systems cultivate fungi (Chaetothyriales) within their domatia, which are fed to larvae. Using Pseudomyrmex penetrator/Tachigali sp. from French Guiana and Petalomyrmex phylax/Leonardoxa africana and Crematogaster margaritae/Keetia hispida, both from Cameroon, as models, we tested the hypothesis that ant-plant-fungus mutualisms co-occur with culturable Actinobacteria. Using selective media, we isolated 861 putative Actinobacteria from the three systems. All C. margaritae/K. hispida samples had culturable Actinobacteria with a mean of 10.0 colony forming units (CFUs) per sample, while 26 % of P. penetrator/Tachigali samples (mean CFUs 1.3) and 67 % of P. phylax/L. africana samples (mean CFUs 3.6) yielded Actinobacteria. The largest number of CFUs was obtained from P. penetrator workers, P. phylax alates, and C. margaritae pupae. 16S rRNA gene sequencing and phylogenetic analysis revealed the presence of four main clades of Streptomyces and one clade of Nocardioides within these three ant-plant mutualisms. Streptomyces with antifungal properties were isolated from all three systems, suggesting that they could serve as protective symbionts, as found in other insects. In addition, a number of isolates from a clade of Streptomyces associated with P. phylax/L. africana and C. margaritae/K. hispida were capable of degrading cellulose, suggesting that Streptomyces in these systems may serve a nutritional role. Repeated isolation of particular clades of Actinobacteria from two geographically distant locations supports these isolates as residents in ant-plant-fungi niches.

Current issues in the evolutionary ecology of ant-plant symbioses.

Ant-plant symbioses involve plants that provide hollow structures specialized for housing ants and often food to ants. In return, the inhabiting ants protect plants against herbivores and sometimes provide them with nutrients. Here, we review recent advances in ant-plant symbioses, focusing on three areas. First, the nutritional ecology of plant-ants, which is based not only on plant-derived food rewards, but also on inputs from other symbiotic partners, in particular fungi and possibly bacteria. Food and protection are the most important 'currencies' exchanged between partners and they drive the nature and evolution of the relationships. Secondly, studies of conflict and cooperation in ant-plant symbioses have contributed key insights into the evolution and maintenance of mutualism, particularly how partner-mediated feedbacks affect the specificity and stability of mutualisms. There is little evidence that mutualistic ants or plants are under selection to cheat, but the costs and benefits of ant-plant interactions do vary with environmental factors, making them vulnerable to natural or anthropogenic environmental change. Thus, thirdly, ant-plant symbioses should be considered good models for investigating the effects of global change on the outcome of mutualistic interactions.

Multi-locus phylogenies of the genus Barteria (Passifloraceae) portray complex patterns in the evolution of myrmecophytism.

The four species of the central African genus Barteria show variation in habitat and in degree of association with ants. Whereas B. solida, restricted to submontane forests, attracts opportunistic ants to extrafloral nectar, the three other species, found in lowland rainforests (B. fistulosa, B. dewevrei) and in littoral scrub (B. nigritana), possess stem domatia of varying shapes and degrees of specialisation, hosting either non-specific arboreal ants (B. nigritana, some B. dewevrei) or two large species of ants of the genus Tetraponera Smith, 1852 that are specific to some species of Barteria (B. fistulosa, some B. dewevrei). We aimed to investigate whether this variation represents an evolutionary trend toward increasing specialisation of mutualism or the reduction or loss of myrmecophytic traits. For this, we determined phylogenetic relationships within the genus using DNA sequences (primarily nuclear ITS) and microsatellite genotypes (11 loci) on a large sample of individuals, mostly from Cameroon and Gabon. The two types of markers support an initial dichotomy that groups B. dewevrei with B. nigritana and B. fistulosa with B. solida respectively. Within these pairs, species do not appear reciprocally monophyletic. At microsatellite loci, B. nigritana forms a clade embedded within B. dewevrei; and within both B. solida and B. fistulosa, geographical populations show levels of differentiation similar to that observed between populations of B. solida and B. fistulosa. Geographic distance alone does not account for genetic differentiation between species, which indicates reproductive isolation. Divergence in each of the two pairs implies evolutionary transitions in habitat and in myrmecophytism. Specialised mutualism with specific ant species of the genus Tetraponera has been lost in species found in more marginal habitats.

Fungi as mutualistic partners in ant-plant interactions.

Associations between fungi and ants living in mutualistic relationship with plants ("plant-ants") have been known for a long time. However, only in recent years has the mutualistic nature, frequency, and geographical extent of associations between tropical arboreal ants with fungi of the ascomycete order Chaetothyriales and Capnodiales (belonging to the so-called "Black Fungi") become clear. Two groups of arboreal ants displaying different nesting strategies are associated with ascomycete fungi: carton-building ants that construct nest walls and galleries on stems, branches or below leaves which are overgrown by fungal hyphae, and plant-ants that make their nests inside living plants (myrmecophytes) in plant provided cavities (domatia) where ants cultivate fungi in small delimited "patches". In this review we summarize the current knowledge about these unsuspected plant-ant-fungus interactions. The data suggest, that at least some of these ant-associated fungi seem to have coevolved with ants over a long period of time and have developed specific adaptations to this lifestyle.

Plant-ants use symbiotic fungi as a food source: new insight into the nutritional ecology of ant-plant interactions.

Usually studied as pairwise interactions, mutualisms often involve networks of interacting species. Numerous tropical arboreal ants are specialist inhabitants of myrmecophytes (plants bearing domatia, i.e. hollow structures specialized to host ants) and are thought to rely almost exclusively on resources derived from the host plant. Recent studies, following up on century-old reports, have shown that fungi of the ascomycete order Chaetothyriales live in symbiosis with plant-ants within domatia. We tested the hypothesis that ants use domatia-inhabiting fungi as food in three ant-plant symbioses: Petalomyrmex phylax/Leonardoxa africana, Tetraponera aethiops/Barteria fistulosa and Pseudomyrmex penetrator/Tachigali sp. Labelling domatia fungal patches in the field with either a fluorescent dye or (15)N showed that larvae ingested domatia fungi. Furthermore, when the natural fungal patch was replaced with a piece of a (15)N-labelled pure culture of either of two Chaetothyriales strains isolated from T. aethiops colonies, these fungi were also consumed. These two fungi often co-occur in the same ant colony. Interestingly, T. aethiops workers and larvae ingested preferentially one of the two strains. Our results add a new piece in the puzzle of the nutritional ecology of plant-ants.

Do host species evolve a specific response to slave-making ants?

BACKGROUND: Social parasitism is an important selective pressure for social insect species. It is particularly the case for the hosts of dulotic (so called slave-making) ants, which pillage the brood of host colonies to increase the worker force of their own colony. Such raids can have an important impact on the fitness of the host nest. An arms race which can lead to geographic variation in host defenses is thus expected between hosts and parasites. In this study we tested whether the presence of a social parasite (the dulotic ant Myrmoxenus ravouxi) within an ant community correlated with a specific behavioral defense strategy of local host or non-host populations of Temnothorax ants. Social recognition often leads to more or less pronounced agonistic interactions between non-nestmates ants. Here, we monitored agonistic behaviors to assess whether ants discriminate social parasites from other ants. It is now well-known that ants essentially rely on cuticular hydrocarbons to discriminate nestmates from aliens. If host species have evolved a specific recognition mechanism for their parasite, we hypothesize that the differences in behavioral responses would not be fully explained simply by quantitative dissimilarity in cuticular hydrocarbon profiles, but should also involve a qualitative response due to the detection of particular compounds. We scaled the behavioral results according to the quantitative chemical distance between host and parasite colonies to test this hypothesis. RESULTS: Cuticular hydrocarbon profiles were distinct between species, but host species did not show a clearly higher aggression rate towards the parasite than toward non-parasite intruders, unless the degree of response was scaled by the chemical distance between intruders and recipient colonies. By doing so, we show that workers of the host and of a non-host species in the parasitized site displayed more agonistic behaviors (bites and ejections) towards parasite than toward non-parasite intruders. CONCLUSIONS: We used two different analyses of our behavioral data (standardized with the chemical distance between colonies or not) to test our hypothesis. Standardized data show behavioral differences which could indicate qualitative and specific parasite recognition. We finally stress the importance of considering the whole set of potentially interacting species to understand the coevolution between social parasites and their hosts.

The cost of myrmecophytism: insights from allometry of stem secondary growth.

BACKGROUND AND AIMS: Plant defence traits against herbivores incur production costs that are usually difficult to measure. However, estimating these costs is a prerequisite for characterizing the plant defence strategy as a whole. Myrmecophytes are plants that provide symbiotic ants with specialized nesting cavities, called domatia, in exchange for protection against herbivores. In the particular case of stem domatia, production of extra wood seems to be the only associated cost, making this indirect defence trait a particularly suitable model for estimating the cost of defence. METHODS: Measurements were made of growth pattern and cumulative production cost of domatia over secondary growth in the myrmecophyte Leonardoxa africana subsp. africana, whose internodes display both a solid basal segment and a hollow distal part (the domatium), thus allowing paired comparison of investment in wood. KEY RESULTS: Previous studies showed that 'overconstruction' of the hollow part of internodes during primary growth is needed for mechanical support. In this study, it is shown that the relationship between the woody cross-sectional area of the solid and hollow parts of internodes is negatively allometric at the beginning of secondary growth and nearly isometric later on. Thus, in hollow stems, the first phase of slow secondary growth compensates for the 'overconstruction' of the ring of wood during primary growth. Moreover, the cumulative production cost of a domatium (estimated as the additional volume of wood required for a hollow stem compared with a solid one) is very high at the beginning of secondary growth and then quickly tends to zero. CONCLUSIONS: Making domatia incurs high costs early in ontogeny, costs that are then amortized later in development of stems and of individual plants. Characterizing ontogenetic variation of the net cost of this peculiar defence mechanism will help us build more accurate theoretical models of resource allocation in myrmecophytes.

Population dynamics of a poultry hematophagous mite: characterization of the population growth and identification of factors of its slowdown using closed mesocosms.

BACKGROUND: A thorough knowledge of the population dynamics of pests and of the main factors affecting population growth is an important prerequisite for the development of effective control strategies. Failures of various treatments aimed at regulating populations of Dermanyssus gallinae are regularly reported in poultry farms and pullulations occur very quickly after first detection. To finely characterize population dynamics of D. gallinae, and to identify the factors modulating population growth, we conducted two successive multi-generation experiments using closed mesocosms equipped with or without automatic counters and housing a host full- or part-time (three nights per week). RESULTS: Population growth was very rapid and the adult to juvenile ratio very different from the prediction by a mathematical model. A male-biased sex ratio was observed in some mesocosms from 21 days and in most mesocosms from 35 days of population growth originating from an inoculum of adult females. A dramatic slowdown in growth was measured in mesocosms equipped with trackers, where the mites' path to the host was constrained. The slowdown in population growth induced by the intermittent presence of the host compared to its full-time presence was much less marked. CONCLUSION: These findings suggest avenues of research for new management methods. They question the relevance of a critical threshold based on traditional trap monitoring to manage D. gallinae. Our results highlight a unique characteristic of D. gallinae that makes it a recalcitrant case to threshold-based practices recommended for integrated pest management (IPM) against other arthropod pests. The dramatic effect of a physical constraint for the mite to access the host (unnatural constrained path) confirms an observation made in 1917 and is a reason to design perches that are less conducive to parasite traffic. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Pollinator and floral odor specificity among four synchronopatric species of Ceropegia (Apocynaceae) suggests ethological isolation that prevents reproductive interference.

Possession of flowers that trap fly pollinators is a conservative trait within the genus Ceropegia, in which pollination systems can be generalized or highly specialized. However, little is known about the role of plant-pollinator interactions in the maintenance of species boundaries. This study examined the degree of plant-pollinator specialization and identified the parameters responsible for specificity among four co-occurring Ceropegia species with overlapping flowering times. All investigated plant species were functionally specialized on pollination by Chloropidae and/or Milichiidae flies and each Ceropegia species was, in turn, ecologically highly specialized on only two pollinating fly morphospecies, though one plant species appeared more generalist. Species-specific fly attraction was due to the differences between plant species in floral scents, floral morphology, colour patterns, and presence of other functional structures, e.g., vibratile trichomes, which were shown to contribute to pollinator attraction in one study species. The combination of these olfactory and visual cues differentially influenced pollinator preferences and thus hindered heterospecific visitation. Furthermore, a pollinator exchange experiment also highlighted that species integrity is maintained through efficient ethological isolation (pollinator attraction). The mechanical isolation mediated by the fit between floral morphology and size and/or shape of fly pollinators appears less pronounced here, but whether or not the morphological match between male (pollinium) and female (guide rails) reproductive organs can impede hybridization remains to be investigated.

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.

Plant-ants feed their host plant, but above all a fungal symbiont to recycle nitrogen.

In ant-plant symbioses, plants provide symbiotic ants with food and specialized nesting cavities (called domatia). In many ant-plant symbioses, a fungal patch grows within each domatium. The symbiotic nature of the fungal association has been shown in the ant-plant Leonardoxa africana and its protective mutualist ant Petalomyrmex phylax. To decipher trophic fluxes among the three partners, food enriched in (13)C and (15)N was given to the ants and tracked in the different parts of the symbiosis up to 660 days later. The plant received a small, but significant, amount of nitrogen from the ants. However, the ants fed more intensively the fungus. The pattern of isotope enrichment in the system indicated an ant behaviour that functions specifically to feed the fungus. After 660 days, the introduced nitrogen was still present in the system and homogeneously distributed among ant, plant and fungal compartments, indicating efficient recycling within the symbiosis. Another experiment showed that the plant surface absorbed nutrients (in the form of simple molecules) whether or not it is coated by fungus. Our study provides arguments for a mutualistic status of the fungal associate and a framework for investigating the previously unsuspected complexity of food webs in ant-plant mutualisms.

Population-level impact of native arthropod predators on the poultry red mite Dermanyssus gallinae.

Stimulating the regulation of pests by their natural enemies is a way to improve the sustainability of agriculture and respect for the environment. However, the presence of natural enemies does not guarantee the existence of a pest control service. To what extent are predatory mites commonly found in henhouses actually able to regulate a major egg industry pest mite, Dermanyssus gallinae? To answer this question, we have experimentally recreated portions of a poultry house ecosystem allowing the development of the pest over several generations in the presence of a chick and detritivorous mites (Astigmata) that are ubiquitous and abundant in layer farms. In these conditions, we compared the growth of D. gallinae populations in the presence and absence of native predatory arthropods. No effect of native predators on the growth of the D. gallinae population could be detected despite high initial predator-to-prey ratios and satisfactory growth of predator populations. Prey switching to the alternative prey Astigmata likely dilutes the effect of predation on the target prey. Further exploration is needed to see whether action could be taken to enhance the effect of top-down regulation.

Predation interactions among henhouse-dwelling arthropods, with a focus on the poultry red mite Dermanyssus gallinae.

BACKGROUND: Analysis of the poorly explored food webs of henhouse-dwelling arthropods would improve biological control against the poultry red mite (PRM) Dermanyssus gallinae (De Geer). This study aimed to identify trophic links among native predatory arthropods, PRM, and alternative preys. In vitro predation tests were carried out to assess (i) the ability of native predators to feed on PRM juvenile and adult stages in two physiological statuses (unfed and freshly blood-fed) in the absence of any physical barrier, (ii) predator preferences between PRM and astigmatic mites, and (iii) predation interactions between PRM predators. RESULTS: Ten arthropod taxa fed on PRM with predation rates ranging from 4% to 95% in our experimental conditions. They belonged to (i) Acari: Androlaelaps casalis (Berlese), Cheyletus spp., Macrocheles muscaedomesticae (Scopoli), M. penicilliger (Berlese), Parasitus fimetorum (Berlese), Dendrolaelaps spp. and Uroobovella fimicola (Berlese); (ii) other Arachnida: Lamprochernes nodosus (Schrank) (Pseudoscorpionida) and a linyphiid spider; and (iii) Insecta: Lyctocoris campestris (Fabricius). These predators varied in their preference for PRM stages and physiological statuses (unfed or freshly blood-fed). When given a choice, most predators preferred to feed on PRM than astigmatic mites. Bidirectional predation occurred within two pairs of PRM predators (M. penicilliger-Lamprochernes nodosus and A. casalis-Cheyletus spp.), and M. penicilliger had a 100% predation rate on A. casalis. CONCLUSION: Our study highlights the potential of various arthropod predators occurring naturally in poultry houses for conservation and augmentative biological control of PRM. Predation interactions between these predators should be accounted for before developing biocontrol agents against PRM. © 2020 Society of Chemical Industry.

Phenotypic plasticity in size of ant-domatia.

Ant-plants produce hollow structures called domatia to host protecting ants. Although size variation in domatia is well documented between related species, intraspecific variation is little explored. The central African ant-plant Barteria dewevrei exibits strong variation in domatium size, giving the opportunity to explore the mechanism underlying variation in a mutualistic trait. We showed that domatium size in Barteria dewevrei varies between sites. We transplanted individual plants between two sites in Gabon where plants have different domatium sizes. Domatium size of transplanted plants changed, revealing that variation in this mutualistic trait is driven by phenotypic plasticity. The two sites differed in their environmental conditions: highland open savanna on sandy soil vs lowland closed tropical rain forest on sandy-loam soil. However, as stomatal density and δ13C of leaves did not differ between sites or between branches produced before and after transplantation, we have no cue on the role of abiotic stress (such as light intensity and water availability) in domatium size variation. As the obligate Tetraponera ant symbionts are too large to fit in the small domatia, variation of the mutualistic trait in response to environmental change through phenotypic plasticity may impact this specialized mutualism.

Evaluating the link between predation and pest control services in the mite world.

Pest regulation by natural enemies has a strong potential to reduce the use of synthetic pesticides in agroecosystems. However, the effective role of predation as an ecosystem service remains largely speculative, especially with minute organisms such as mites.Predatory mites are natural enemies for ectoparasites in livestock farms. We tested for an ecosystem level control of the poultry pest Dermanyssus gallinae by other mites naturally present in manure in poultry farms and investigated differences among farming practices (conventional, free-range, and organic).We used a multiscale approach involving (a) in vitro behavioral predation experiments, (b) arthropod inventories in henhouses with airborne DNA, and (c) a statistical model of covariations in mite abundances comparing farming practices.Behavioral experiments revealed that three mites are prone to feed on D. gallinae. Accordingly, we observed covariations between the pest and these three taxa only, in airborne DNA at the henhouse level, and in mites sampled from manure. In most situations, covariations in abundances were high in magnitude and their sign was positive.Predation on a pest happens naturally in livestock farms due to predatory mites. However, the complex dynamics of mite trophic network prevents the emergence of a consistent assemblage-level signal of predation. Based on these results, we suggest perspectives for mite-based pest control and warn against any possible disruption of ignored services through the application of veterinary drugs or pesticides.

Ant-plants and fungi: a new threeway symbiosis.

Symbioses between plants and fungi, fungi and ants, and ants and plants all play important roles in ecosystems. Symbioses involving all three partners appear to be rare. Here, we describe a novel tripartite symbiosis in which ants and a fungus inhabit domatia of an ant-plant, and present evidence that such interactions are widespread. We investigated 139 individuals of the African ant-plant Leonardoxa africana for occurrence of fungus. Behaviour of mutualist ants toward the fungus within domatia was observed using a video camera fitted with an endoscope. Fungi were identified by sequencing a fragment of their ribosomal DNA. Fungi were always present in domatia occupied by mutualist ants but never in domatia occupied by opportunistic or parasitic ants. Ants appear to favour the propagation, removal and maintenance of the fungus. Similar fungi were associated with other ant-plants in Cameroon. All belong to the ascomycete order Chaetothyriales; those from L. africana formed a monophyletic clade. These new plant-ant-fungus associations seem to be specific, as demonstrated within Leonardoxa and as suggested by fungal phyletic identities. Such tripartite associations are widespread in African ant-plants but have long been overlooked. Taking fungal partners into account will greatly enhance our understanding of symbiotic ant-plant mutualisms.

Transmission of fungal partners to incipient Cecropia-tree ant colonies.

Ascomycete fungi in the nests of ants inhabiting plants (= myrmecophytes) are very often cultivated by the ants in small patches and used as food source. Where these fungi come from is not known yet. Two scenarios of fungus recruitment are possible: (1) random infection through spores or hyphal fragments from the environment, or (2) transmission from mother to daughter colonies by the foundress queen. It is also not known at which stage of the colony life cycle fungiculture is initiated, and whether the- symbiont fungi serve as food for the ant queen. To clarify these questions, we investigated four Azteca ant species inhabiting three different Cecropia species (C. insignis, C. obtusifolia, and C. peltata). We analysed an rRNA gene fragment from 52 fungal patches produced by founding queens and compared them with those from established Azteca colonies (n = 54). The infrabuccal pockets of winged queens were dissected to investigate whether young queens carry fungi from their mother colony. Additionally, 15N labelling experiments were done to verify whether the queen feeds on the patches until she is nourished by her first worker offspring. We infer from the results that the fungi cultivated in hollow plant structures are transferred from the parental colony of the young queen. First, fungal genotypes/OTU diversity was not significantly different between foundress queen patches and established colonies, and second, hyphal parts were discovered in the infrabuccal pockets of female alates. We could show that fungiculture already starts before queens lay their eggs, and that the queens do not feed on fungal patch material but feed it to the larvae. Our findings suggest that fungiculture may be crucial for successful colony founding of arboreal ants in the tropics.

The unique functioning of a pre-Columbian Amazonian floodplain fishery.

Archaeology provides few examples of large-scale fisheries at the frontier between catching and farming of fish. We analysed the spatial organization of earthen embankments to infer the functioning of a landscape-level pre-Columbian Amazonian fishery that was based on capture of out-migrating fish after reproduction in seasonal floodplains. Long earthen weirs cross floodplains. We showed that weirs bear successive V-shaped features (termed 'Vs' for the sake of brevity) pointing downstream for outflowing water and that ponds are associated with Vs, the V often forming the pond's downstream wall. How Vs channelled fish into ponds cannot be explained simply by hydraulics, because Vs surprisingly lack fishways, where, in other weirs, traps capture fish borne by current flowing through these gaps. We suggest that when water was still high enough to flow over the weir, out-migrating bottom-hugging fish followed current downstream into Vs. Finding deeper, slower-moving water, they remained. Receding water further concentrated fish in ponds. The pond served as the trap, and this function shaped pond design. Weir-fishing and pond-fishing are both practiced in African floodplains today. In combining the two, this pre-Columbian system appears unique in the world.