Influence of caffeine on the survival of leaf-cutting ants Atta sexdens rubropilosa and in vitro growth of their mutualistic fungus

BACKGROUND: Leaf-cutting ants collect plant fresh material for the cultivation of their mutualistic fungus. Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) cause great economic losses through their foraging activity, mainly in agriculture. The main control method is the application of granulated toxic baits incorporated with an active ingredient (AI). The present goal is to evaluate the effect of caffeine on in vitro growth of the mutualistic fungus and on the survival of the leaf-cutting ants, aiming to verify the potential toxicity of this secondary metabolite over these organisms. RESULTS: Three distinct patterns of fungal growth correlated with caffeine concentration were observed: (1) no effect (0.01% caffeine); (2) intermediate growth reduction (0.05% caffeine); (3) drastic growth reduction (0.10 and 0.50% caffeine). The highest caffeine concentration causes fungus death in the first week. As for insect survival, caffeine does not seem to exert any effect. The treatments with diet containing caffeine showed similar values of M50, irrespective of caffeine concentration. CONCLUSION: As caffeine was shown to reduce growth of the mutualistic fungus of Atta sexdens rubropilosa, but with no conclusive effect on insect survival, a hypothetical explanation for the selection of different Coffea species by this leaf-cutting ant species might be associated with caffeine toxicity to the fungus. Copyright (C) 2011 Society of Chemical Industry

Considerations for Assessing Maximum Critical Temperatures in Small Ectothermic Animals: Insights from Leaf-Cutting Ants

The thermal limits of individual animals were originally proposed as a link between animal physiology and thermal ecology. Although this link is valid in theory, the evaluation of physiological tolerances involves some problems that are the focus of this study. One rationale was that heating rates shall influence upper critical limits, so that ecological thermal limits need to consider experimental heating rates. In addition, if thermal limits are not surpassed in experiments, subsequent tests of the same individual should yield similar results or produce evidence of hardening. Finally, several non-controlled variables such as time under experimental conditions and procedures may affect results. To analyze these issues we conducted an integrative study of upper critical temperatures in a single species, the ant Atta sexdens rubropiosa, an animal model providing large numbers of individuals of diverse sizes but similar genetic makeup. Our specific aims were to test the 1) influence of heating rates in the experimental evaluation of upper critical temperature, 2) assumptions of absence of physical damage and reproducibility, and 3) sources of variance often overlooked in the thermal-limits literature; and 4) to introduce some experimental approaches that may help researchers to separate physiological and methodological issues. The upper thermal limits were influenced by both heating rates and body mass. In the latter case, the effect was physiological rather than methodological. The critical temperature decreased during subsequent tests performed on the same individual ants, even one week after the initial test. Accordingly, upper thermal limits may have been overestimated by our (and typical) protocols. Heating rates, body mass, procedures independent of temperature and other variables may affect the estimation of upper critical temperatures. Therefore, based on our data, we offer suggestions to enhance the quality of measurements, and offer recommendations to authors aiming to compile and analyze databases from the literature.

Sodium-specific foraging by leafcutter ant workers (Atta cephalotes, Hymenoptera: Formicidae)

1. Sodium is often a limiting nutrient for terrestrial animals, and may be especially sought by herbivores. Leafcutter ants are dominant herbivores in the Neotropics, and leafcutter foraging may be affected by nutritional demands of the colony and/or the demands of their symbiotic fungal mutualists. We hypothesized that leafcutter colonies are sodium limited, and that leafcutter ants will therefore forage specifically for sodium. 2. Previous studies demonstrated that leafcutter Atta cephalotes Linnaeus workers preferentially cut and remove paper baits treated with NaCl relative to water control baits. Atta cephalotes colonies in this study were presented with baits offering NaCl, Na2SO4, and KCl to test whether leafcutters forage specifically for sodium. Sucrose and water were used as positive and negative controls, respectively. 3. Atta foragers removed significantly more of the baits treated with NaCl and Na2SO4 than the KCl treatment, which did not differ from water. The NaCl and Na2SO4 treatments were collected at similar rates. We conclude A. cephalotes forage specifically for sodium rather than for anions (chloride) or solutes in general. This study supports the hypothesis that leafcutter ants are limited by, and preferentially forage for, sodium.

Morphological patterns and community organization in leaf-litter ant assemblages

We present a new approach to determine the number and composition of guilds, using the hyperdiverse leaf-litter ant fauna as a model, based on appropriate morphological variables and species co-occurrence null models to describe the complex assemblages of interacting Species Community structure at the 1-m(2) scale. We obtained 18 linear morphometric measures from 949 workers of 171 leaf-litter ant species (18762 measurements) surveyed in four Atlantic Forest localities to test whether the assemblages are morphologically structured; the morphological characters were selected to indicate diet and foraging habits. Principal components analysis was used to characterize the morphospace and to describe the guild structure (number of species and composition). The guild proportionality assembly rule (significant tendency toward constant proportion of species in guilds) was assessed at the 1-m(2) scale. Our analysis indicates that the division of leaf-litter ants into guilds is based mainly on microhabitat distribution in the leaf-litter, body size and shape, eye size, and phylogeny. The same guild scheme applied to four more sites shows that different Atlantic Forest areas have the same leaf-fitter ant guilds. The guild proportionality assembly rule was confirmed for most guilds, Suggesting that there are guild-specific limitations on species coexistence within assemblages; on the other hand, in a few cases the variance in guild proportion was greater than expected under the null assumptions. Other studies on ant functional group classification are partially supported by our quantitative morphological analysis. Our results, however, imply that there are more compartments than indicated in previous models, particularly among cryptic species (confined to soil and litter) and tropical climate specialists. We argue that a general null model for the analysis of species association based oil morphology can reveal objectively defined groups and may thus contribute to a robust theory to explain community structure in general and have important consequences on studies of litter ant community ecology in particular.

Natural history of an ant-plant-butterfly interaction in a Neotropical savanna

Non-myrmecophilous lepidopteran larvae using plants bearing ant attractants such as extrafloral nectaries are good models for studying morphological and behavioural mechanisms against ant predation. Udranomia spitzi (Hesperiidae) is a butterfly whose larvae feed on leaves of Ouratea spectabilis (Ochnaceae), a plant with extrafloral nectaries. We described the early stages of U. spitzi, and used field observations and experiments to investigate the defensive strategies of caterpillars against predatory ants. Larvae pass through five instars and pupation occurs inside larval leaf shelters. Ant-exclusion experiments revealed that the presence of ants did not affect significantly caterpillar survival. Predation experiments showed that vulnerability to ant predation decreased with increase in larval size. The present study showed that predatory ants are not as relevant as demonstrated for other systems, and also illustrates how observational data and field experiments can contribute to a better understanding of the biology and ecology of a species of interest.

A new ant species of Oxyepoecus (Hymenoptera: Formicidae: Myrmicinae), with the description of Oxyepoecus browni gyne and new records for the genus

We describe a new species of Oxyepoecus Santschi, 1926, Oxyepoecus regularis sp. nov., based on workers and a gyne collected in "Caatinga Arbórea" (Arboreal Shrubland) in Milagres and "Mata Seca" (Dry Forest) in Boa Vista do Tupim, both in the state of Bahia, Brazil. The gyne of Oxyepoecus browni Albuquerque & Brandão, 2004, collected in the same leaf litter ant survey, is also described. In addition, we present new records for Oxyepoecus species in Northeastern Brazil.

Molecular characterization of a mariner-like element in the Atta sexdens rubropilosa genome

Mobile elements are widely present in eukaryotic genomes. They are repeated DNA segments that are able to move from one locus to another within the genome. They are divided into two main categories, depending on their mechanism of transposition, involving RNA (class I) or DNA (class II) molecules. The mariner-like elements are class II transposons. They encode their own transposase, which is necessary and sufficient for transposition in the absence of host factors. They are flanked by a short inverted terminal repeat and a TA dinucleotide target site, which is duplicated upon insertion. The transposase consists of two domains, an N-terminal inverted terminal repeat binding domain and a C-terminal catalytic domain. We identified a transposable element with molecular characteristics of a mariner-like element in Atta sexdens rubropilosa genome. Identification started from a PCR with degenerate primers and queen genomic DNA templates, with which it was possible to amplify a fragment with mariner transposable-element homology. Phylogenetic analysis demonstrated that this element belongs to the mauritiana subfamily of mariner-like elements and it was named Asmar1. We found that Asmar1 is homologous to a transposon described from another ant, Messor bouvieri. The predicted transposase sequence demonstrated that Asmar1 has a truncated transposase ORF. This study is part of a molecular characterization of mobile elements in the Atta spp genome. Our finding of mariner-like elements in all castes of this ant could be useful to help understand the dynamics of mariner-like element distribution in the Hymenoptera.

Tropidomyrmex elianae, a new myrmicine ant genus and species from Brazil, tentatively assigned to Solenopsidini (Hymenoptera, Formicidae)

A new myrmicine ant, Tropidomyrmex elianae gen. n. & sp. n., is described from southeastern and central Brazil, based on workers, ergatoid gynes, males and larvae. Tropidomyrmex workers are relatively small, monomorphic, characterized mainly by the feebly pigmented and extremely thin integument; subfalcate mandibles bearing a single apical tooth; palpal formula 1,2; clypeus relatively broad and convex; reduced compound eyes; propodeum unarmed and with a strongly medially depressed declivous face; double and bilobed well developed subpostpetiolar processes; and peculiarities in the sting apparatus. A colony fragment of T. elianae containing workers, ergatoid gynes, males, and brood was found inside a ground termite nest (Anoplotermes pacificus Apicotermitinae) in a montane rocky scrubland in the state of Minas Gerais, southeastern Brazil. Tropidomyrmex elianae is known also from two workers collected in leaf litter samples processed with a Winkler extractor, from the state of Tocantins, central-north Brazil. Despite the differences from the accepted solenopsidine genera, Tropidomyrmex is tentatively assigned to this tribe. Within the solenopsidine ants, the genus is apparently related to Tranopelta. Tropidomyrmex is marked by extreme reductions, perhaps reflecting adaptations to particular habits and habitats.

A taxonomic revision of the Neotropical myrmicine ant genus Lachnomyrmex wheeler (Hymenoptera : Formicidae)

The Neotropical ant genus Lachnomyrmex Wheeler, 1910 is revised for the first time. A revised generic description is provided for workers and gynes; males remain unknown. Morphological patterns combined with geographical data led to the recognition of 16 species, of which 10 are here described as new: L. amazonicus sp. n., L. fernandezi sp. n., L. grandis Fernandez & Baena, L. haskinsi Smith, L. lattkei sp. n., L. longinodus Fernandez & Baena, L. longinoi sp. n., L. mackayi sp. n., L. nordestinus sp. n., L. pilosus Weber, L. laticeps sp. n., L. platynodus sp. n., L. plaumanni Borgmeier, L. regularis sp. n., L. scrobiculatus Wheeler, and L. victori sp. n. The gynes of L. grandis, L. haskinsi, and L. pilosus are described for the first time. Illustrated identification keys for workers, distribution maps, and high resolution illustrations are supplied for all species. Some Lachnomyrmex species are relatively common in the leaf-litter of submontane wet forests in Central and South America; others remain known by very few individuals. The relatively small and apparently monogynic Lachnomyrmex colonies inhabit small nests in the ground, among rotten leaves and inside fallen logs. Workers generally forage alone on the ground or within the leaf litter, but have been recorded also on tree trunks, and apparently do not recruit nestmates.

Revisionary studies on the enigmatic neotropical ant genus Stegomyrmex Emery, 1912 (Hymenoptera : Formicidae : Myrmicinae), with the description of two new species

The recent increase in leaf litter ants sampling effort in Neotropical wet forests has revealed new and interesting records of the highly specialized myrmicine ant genus Stegomyrmex Emery, previously considered as extremely rare. We present a modified diagnosis for the genus and describe Stegomyrmex bensoni n. sp. and S. olindae n. sp., based on, respectively, workers, males, and gyne (central-north Brazil) and on a single worker (northern Brazil). Stegomyrmex vizottoi Diniz (southeastern Brazil) is redescribed and compared with S. olindae n. sp.; these species present significant differences in size, sculpturation, and sting apparatus morphology. The males of S. vizottoi are described for the first time. A key for workers and queens and a distribution map for the five know Stegomyrmex species are provided.

Assembly of Myrmelachista Roger (Formicidae: Formicinae) in twigs fallen on the leaf litter of Brazilian Atlantic Forest

The nests of Myrmelachista are found in tree trunk cavities and branches. The biology of these arboreal ants is still relatively unknown. We investigated the nesting behaviour of this genus in fallen dry twigs in the Atlantic rainforest of southeastern Brazil. Physical characteristics of the nests, colony demographics, presence of breeders and ant worker sizes were recorded. Samples were collected weekly for 12 months, along open, sunny and undisturbed trails within forest remnants. In all, 202 nests were collected and six species were recorded. Myrmelachista ruszkii had the highest population of immatures and the greatest number of nests found. Myrmelachista nodigera had the smallest workers, was the least populous and the colony was housed in the finest branches. These results expand the current knowledge about the diversity and biology of Myrmelachista in the Brazilian Atlantic forest, and describe polydomic nests and competitive behaviour of M. ruszkii.

PHYSIOLOGICAL CHANGES IN SEEDS AND ANTIOXIDANT METABOLISM OF LETTUCE SEEDLING EXPOSED TO THE ACTION OF Zantedeschia aethiopica Spreng. LEAF EXTRACT

This work aimed to evaluate the influence of different concentrations of Zantedeschia aethiopica Spreng. extract on the physiological performance of the seed and on the response of the antioxidant metabolism of lettuce seedlings. The treatments consisted of leaves extracts from Z. aethiopica at concentrations of 0, 6, 12, 25 and 50%. Germination, first germination count, germination speed and index, length of shoot and radicle, seedling total dry mass, chlorophyll content, activity of superoxide dismutase, catalase and ascorbarte peroxidase enzymes, lipid peroxidation, hydrogen peroxide quantification and seedling emergence, length of organs, and total dry mass of seedlings were evaluated. The percentage of germination, the length of the shoot and radicle of seedlings and the total dry mass of seedlings grown in the greenhouse were reduced as the concentration of the extract increased. There were increases of electrical conductivity, of superoxide dismutase, catalase and ascorbate peroxidadase enzymes and the amount of hydrogen peroxide and lipid peroxidation in seedlings with increasing extract concentration. The extract reduced the physiological quality of lettuce seeds and induced an increased production of hydrogen peroxide in seedlings, which increased the activity of antioxidant enzymes that were not effective in tissue detoxification, resulting in cellular damage and increased numbers of abnormal seedlings.

Characterization of the procera Tomato Mutant Shows Novel Functions of the SlDELLA Protein in the Control of Flower Morphology, Cell Division and Expansion, and the Auxin-Signaling Pathway during Fruit-Set and Development

procera (pro) is a tall tomato (Solanum lycopersicum) mutant carrying a point mutation in the GRAS region of the gene encoding SlDELLA, a repressor in the gibberellin (GA) signaling pathway. Consistent with the SlDELLA loss of function, pro plants display a GA-constitutive response phenotype, mimicking wild-type plants treated with GA(3). The ovaries from both nonemasculated and emasculated pro flowers had very strong parthenocarpic capacity, associated with enhanced growth of preanthesis ovaries due to more and larger cells. pro parthenocarpy is facultative because seeded fruits were obtained by manual pollination. Most pro pistils had exserted stigmas, thus preventing self-pollination, similar to wild-type pistils treated with GA(3) or auxins. However, Style2.1, a gene responsible for long styles in noncultivated tomato, may not control the enhanced style elongation of pro pistils, because its expression was not higher in pro styles and did not increase upon GA(3) application. Interestingly, a high percentage of pro flowers had meristic alterations, with one additional petal, sepal, stamen, and carpel at each of the four whorls, respectively, thus unveiling a role of SlDELLA in flower organ development. Microarray analysis showed significant changes in the transcriptome of preanthesis pro ovaries compared with the wild type, indicating that the molecular mechanism underlying the parthenocarpic capacity of pro is complex and that it is mainly associated with changes in the expression of genes involved in GA and auxin pathways. Interestingly, it was found that GA activity modulates the expression of cell division and expansion genes and an auxin signaling gene (tomato AUXIN RESPONSE FACTOR7) during fruit-set.

Social facilitation and food partitioning in the queenless ant Dinoponera quadriceps (Hymenoptera: Formicidae)

Social facilitation occurs when an animal is more likely to behave in a certain way in response to other animals engaged in the same behaviour. For example, an individual returning to the nest with food stimulates other ants to leave and to forage. In the present study we demonstrate the existence of new facets in the colony organization of Dinoponera quadriceps: a positive feedback between the incoming food and the activation of new foragers, and the occurrence of incipient task partitioning during the food sharing. Lower-ranked workers located inside the nest process protein resources and higher-ranked workers handle smaller pieces and distribute them to the larvae. In conclusion, D. quadriceps has a decentralized pattern of task allocation with a double regulatory mechanism, which can be considered a sophisticated aspect of division of labour in ponerine ants.

Assessment of the high-dose concept and level of control provided by MON 87701 x MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil

BACKGROUND: Genetically modified MON 87701 X MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP-synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high-dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS: The purified Cry1Ac protein was more active against A. gemmatalis [LC50 (FL 95%) = 0.23 (0.150.34) mu g Cry1Ac mL-1 diet] than P. includens [LC50 (FL 95%) = 3.72 (2.654.86) mu g Cry1Ac mL-1 diet]. In bioassays with freeze-dried MON 87701X MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf-disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701X MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS: The MON 87701X MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high-dose event only to A. gemmatalis. Copyright (c) 2012 Society of Chemical Industry