Energy substrate used by workers of leaf-cutting ants during nest excavation

Energy substrate used by workers of leaf-cutting ants during nest excavation. In this study we aimed to ascertain whether leaf-cutting ant workers lose body reserves (fat or sugars) as a function of nest excavation. For each treatment, we isolated 10 workers of Atta sexdens into two experimental groups, Control (C- without excavation) and Soil (S- with excavation), which were kept for different time intervals (0, 24, 48 or 72 hours), totaling 700 tested workers. We then determined the concentration of soluble carbohydrates and total lipid content in them. The total carbohydrates were determined colorimetrically, based on the reaction between carbohydrates and sulfuric acid-phenol. For determination of lipids, the insects were immersed in organic solvent until they reached a constant weight. Our results showed that carbohydrates are consumed during nest excavation activities. In the experimental groups S24, S48 and S72, there was an average reduction of 5.82 (20.42%), 14.31 (44.96%) and 13.27 (43.96%) µ.mg-1 in soluble sugar when compared with the experimental groups that did not excavate. Furthermore, the lipids were not used during this activity. With respect to dry mass of the workers, their values were C0 = 8%, C24 = 10.4%, C48 = 9.2%, C72 = 10%, S24 = 9.2%, S48 = 8.7% and S72 = 8.5%. Our results show experimentally that the source of energy for nest excavation is carbohydrates, whereas lipids are conserved for other activities.

Preparation of mitotic chromosomes of leaf-cutting ants from the genera Atta and Acromyrmex

Some modifications were made to the methodology of Imai et al. (Jpn. J. Genet. 63: 159-185, 1988) for cytogenetic analysis of the leaf-cutting ants Atta sexdens piriventris and Acromyrmex heyeri (Hymenoptera, Formicidae), shortening preparation time and improving chromosomal preparations. The brain ganglia of prepupae were dissected in a 0.0025% hypotonic solution of colchicine, placed on a glass slide on a cold plate (4 ± 1oC) for 20 min. The material was fixed directly on the cold slide (with cold fixative I), macerated with a histological needle and fixed again with fixative I, followed by fixatives II and III, all of them cold. The slide was flame-dried right after the use of fixative III, and it was allowed to air-dry at room temperature for 2 h. The resulting metaphases presented less contracted chromosomes, with separated and well defined sister chromatids at a high frequency, when the material was processed in the manner described and stained with 3% Giemsa in phosphate buffer (pH 6.8) for 15 min.

Low variation in ribosomal DNA and internal transcribed spacers of the symbiotic fungi of leaf-cutting ants (Attini: Formicidae)

Leaf-cutting ants of the genera Atta and Acromyrmex (tribe Attini) are symbiotic with basidiomycete fungi of the genus Leucoagaricus (tribe Leucocoprineae), which they cultivate on vegetable matter inside their nests. We determined the variation of the 28S, 18S, and 5.8S ribosomal DNA (rDNA) gene loci and the rapidly evolving internal transcribed spacers 1 and 2 (ITS1 and ITS2) of 15 sympatric and allopatric fungi associated with colonies of 11 species of leafcutter ants living up to 2,600 km apart in Brazil. We found that the fungal rDNA and ITS sequences from different species of ants were identical (or nearly identical) to each other, whereas 10 GenBank Leucoagaricus species showed higher ITS variation. Our findings suggest that Atta and Acromyrmex leafcutters living in geographic sites that are very distant from each other cultivate a single fungal species made up of closely related lineages of Leucoagaricus gongylophorus. We discuss the strikingly high similarity in the ITS1 and ITS2 regions of the Atta and Acromyrmex symbiotic L. gongylophorus studied by us, in contrast to the lower similarity displayed by their non-symbiotic counterparts. We suggest that the similarity of our L. gongylophorus isolates is an indication of the recent association of the fungus with these ants, and propose that both the intense lateral transmission of fungal material within leafcutter nests and the selection of more adapted fungal strains are involved in the homogenization of the symbiotic fungal stock.

Levels of leaf Herbivory in Amazonian trees from different stages In forest regeneration

Leaves from 120 canopy trees and 60 understory tree saplings growing in primary and secondary forests near Manaus, Brazil, were collected for determination of standing levels of herbivory (percent leaf area lost). Overall, levels of herbivory on leaves of central Amazonian trees were low. About one quarter of the leaves examined (n = 855) had no damage at all. In most other Neotropical sites studied the mean percentage of herbivory was found to vary between 5.7 and 13.1%, whereas in Manaus it was only 3.1%. The data presented here support the contention that levels of herbivore damage are positively related to soil fertility. No significant difference was found in herbivory levels between canopy trees and understory saplings. Also, there was no difference in damage between leaves from pioneer and late successional trees. Field assays of preference, however, revealed that leaves from pioneer trees are more palatable to leaf-cutting ants (Atta laevigata). This effect was dependent upon leaf age, being observed in mature leaves, but not in young leaves. The greater rate of leaf production in secondary forests may be a factor accounting for the greater abundance of leaf-cutting ants in secondary compared to primary forests.

Seasonality, age structure and reproduction of Leptodactylus (Lithodytes) lineatus (Anura, Leptodactylidae) in Rondônia state, southwestern Amazon, Brazil

Leptodactylus(Lithodytes) lineatus (Schneider, 1799) is an Amazonian leaf litter frog considered rare or uncommon in several studies on anuran communities. Despite being a widely distributed frog in Amazonian forests, knowledge of the biology and ecology of this species is relatively scarce. This species has been reported to live in association with leaf-cutter ant nests (Atta spp.) during the breeding period. In this paper we present data on the seasonality of this species and some reproductive information gathered at a locality of Rondônia state, northwestern Brazil. Field work was carried out between April 2001 and March 2002, with the use of pitfall traps with drift fences as a survey method. Leptodactylus (L.) lineatus had a higher capture frequency in this locality compared to that of other studies carried out in other Amazonian localities, possibly because this species has secretive habits, such as calling and breeding from nests of leaf-cutting ants, and are difficult to find during visual encounter surveys. The breeding period occurs between October and March. Calling males and egg-bearing females were found between September and February and juvenile recruitment occurred mainly from the end of the rainy season to the beginning of the dry season (February to June). Males and females show sexual dimorphism in SVL, females being significantly larger than males. The number of ovarian eggs per female varies from 110 to 328 and analyses indicate that there is a significant correlation with SVL.

Emigration of a colony of the leaf-cutting ant Acromyrmex heyeri Forel (Hymenoptera, Formicidae)

Emigration of a colony of the leaf-cutting ant Acromyrmex heyeri Forel (Hymenoptera, Formicidae). Colony migration is a poorly studied phenomenon in leaf-cutting ants. Here we report on the emigration of a colony of the leaf-cutting ant A. heyeri in Brazil. The colony emigrated to a new location 47.4 m away from the original nest site, possibly because it had undergone considerable stress due to competitive interactions with a colony of Acromyrmex crassispinus.

Protein content of leaf-cutting ant queens before the nuptial flight and during the post-claustral phase

Protein content of leaf-cutting ant queens before the nuptial flight and during the post-claustral phase. This study evaluated the crude protein content of queens of Atta sexdens before the nuptial flight and after the claustral phase in laboratory and field colonies. The hypothesis was that protein is used for survival of the queen and for early colony growth during the claustral phase. Additionally, the nest morphology, live biomass and adult population of field colonies were evaluated. Crude protein was determined by digestion of the organic material with sulfuric acid at high temperatures. The mean crude protein content was 123.23 ± 11.20 mg for females before the nuptial flight and 70.44 ± 12.21 mg for laboratory-reared queens after the claustral phase. The post-claustral crude protein content of field-collected queen was 55.90 ± 9.18 mg. With respect to the loss of crude protein as a function of duration of the claustral phase, laboratory-reared queens lost 52.79 mg and field-collected queens lost 67.33 mg compared to females before the nuptial flight. A positive linear correlation was observed between the weight of field-collected queens (256.4 ± 36.3 mg) and colony biomass (13.02 ± 9.12 g), but there was no correlation between biomass and nest depth (13.11 ± 3.82 cm). As expected, the present results support the hypothesis that protein is used for survival of the queen and for early colony growth, as demonstrated by the reduction in crude protein content as a function of duration of the claustral phase. To our knowledge, this is the first study to provide data of the dynamics of protein reserves in leaf-cutting ant queens during the claustral phase.

Leaf-cutting ant attack in initial pine plantations and growth of defoliated plants

The objective of this work was to evaluate the natural attack by Acromyrmex crassispinus in initial Pinus taeda plantations without control measures against ants, as well as the effect of defoliation in seedlings of P. taeda. Evaluations of the attack of leaf-cutting ants on P. taeda plantations were done monthly in the first six months, then 9 and 12 months after planting. The percentages of plants that were naturally attacked by ants were registered. The effect of defoliation was evaluated by artificial defoliation, simulating the natural patterns of attack by A. crassispinus on P. taeda seedlings. The natural attack of A. crassispinus was greater during the first months after planting, being more intense in the first 30 days. Artificial defoliation indicated that there were no significant losses in diameter and height in plants with less than 75% defoliation. However, there were significant losses in diameter and height in plants with 100% defoliation, independently of the cut of the apical meristem, and also plant death. The control of leaf-cutting ants in P. taeda plantings, in which A. crassispinus is the most frequent leaf-cutting ant, should be intense only at the beginning of planting, since the most severe attacks occur during this time.

Harvesting of Protium heptaphyllum (Aubl.) March. seeds (Burseraceae) by the leaf-cutting ant Atta sexdens L. promotes seed aggregation and seedling mortality

The role played by leaf-cutting ants as seed dispersers of non-myrmecochorous plants remains poorly understood. Here we document the harvesting of Protium heptaphyllum (Aubl.) March. seeds (Burseraceae) by the leaf-cutting ant Atta sexdens L. and its consequences for (1) seed deposition pattern; (2) seed germination; and (3) seedling mortality. The study was carried out at Dois Irmãos, a 390 ha reserve of Atlantic forest, northeast Brazil. Ant-seed harvesting on the ground was detected in 18.5% of all fruiting trees and ants harvested 41.1% ± 19.7% of the seed crop (mean ± s). In average, ants piled seeds 3.4 ± 2.2 m away from the trunk of parent trees and seed density in these piles reached 128.8 ± 138.8 seeds 0.25 m² during the peak of seed discarding by ants. During a 13 month period, mean seedling mortality varied from 0.54% up to 10.6% in ant-made seed piles vs. 0.05-4.2% in control samples, what resulted in a total seedling mortality of 97.7% vs. 81%. Ants systematically cut seedling epicotyls, accounting for 55% of seedling mortality in seed piles, whereas only 14 seedlings (4.2%) were cut by ants in the control samples. Our results suggest that seed harvesting by A. sexdens (1) affects approximately 20% of fruiting P. heptaphyllum trees and their seed crops; (2) promotes short-distance seed dispersal and high levels of seed aggregation; and (3) reduces seedling survival beneath parents.

The diet of the black widow spider Latrodectus mirabilis (Theridiidae) in two cereal crops of central Argentina

The spider Latrodectus mirabilis (Holmberg, 1876) is commonly found in cereals crops of central Argentina. We studied its diet composition at the field and capture rate on leaf-cutting ants based on laboratory experiments. This study comprises the first approach that documents the diet of L. mirabilis in wheat and oat fields of central Argentina. We identified 1,004 prey items collected from its webs during the last phenological stages of both cereal crops. The prey composition was variable but the spiders prey mainly on ants (Formicidae, Hymenoptera), who represented more than 86% of the total. Meanwhile, in the capture rate experiences we registered a high proportion of ants captured by spiders at the beginning of experiences, capturing the half of the ants from total in the first four hours. Summarizing, we reported a polyphagous diet of this spider species in wheat and oat fields. Ants were the most important prey item of this spider, as found in other Latrodectus spiders around the world.

Insects associated with tropical foliage produced in the coffee growing region of Colombia

We conducted a survey of insects and pest management practices on 34 farms growing ornamental tropical foliage plants in the central coffee region of Colombia over two years. Tropical foliage provided habitat for a diverse range of insects. In total, phytophagous or detritivorous insects from six orders, 40 families and 62 genera were collected. The most common were Hemiptera (29 genera from 16 families), followed by Coleoptera (17 genera from 4 families), Diptera (5 genera from 5 families), Lepidoptera (5 genera from 4 families), Hymenoptera (3 genera from 2 families) and Orthoptera (2 genera from 2 families). The most common phytophagous species were leaf cutting ants (Atta and Acromyrmex spp.), leaf beetles (Chrysomelidae), leafhoppers (Cicadellidae), stinkbugs (Pentatomidae), squash bugs (Coreidae), tree hoppers (Membracidae) and plant hoppers (Fulgoridae). Beneficial insects identified from tropical foliage included predators and parasitoids amongst 5 orders, 12 families and 22 genera. The most abundant were predators among the Coccinellidae, Chrysopidae, Reduviidae, Lycidae and Formicidae but only low numbers of parasitoids (Ichneumonidae, Braconidae and Tachinidae) were collected. A pest management questionnaire given to growers revealed a preponderance of reliance on broad spectrum insecticides with a smaller number of growers (approximately one third) also using some biological control methods. Our survey contributes basic information regarding diversity of Neotropical insects associated with ornamental foliage plants.

Dynamics of the restoration of physical trails in the grass-cutting ant Atta capiguara (Hymenoptera, Formicidae)

ABSTRACT Dynamics of the restoration of physical trails in the grass-cutting ant Atta capiguara. Leaf-cutting ants of the genus Atta build long physical trails by cutting the vegetation growing on the soil surface and removing the small objects they find across their path. Little is known on the dynamics of trail construction in these ants. How much time do they need to build a trail? To answer this question we selected six trails belonging to two different nests of A. capiguara and removed on each trail a block of soil of 20 cm × 15 cm that included a portion of the physical trail. This block was then replaced by a new block of the same size that was removed in the pasture near the trail and that was uniformly covered by the same type of vegetation as that found on the block of soil that was removed. The time required to restore the trail was then evaluated by the length of the grass blades found along the former location of the trail. The results show that ants rapidly restore the portion of the physical trail that was interrupted, which suggests that they could also do the same after their trails have been recolonized by the vegetation.

Spittlebug Cephisus siccifolius damaging eucalypt plants in the State of Bahia, Brazil

Most common pests of eucalyptus plants in Brazil are leaf cutting ants and defoliating caterpillars. Other groups, eventually, feeding on eucalyptus include Heteroptera and cicadas. The objectives of this work were to identify and to describe attack symptoms of Cephisus siccifolius (Walker 1851) (Hemiptera: Aphrophoridae) on eucalyptus trees in a stand of 25.77 hectares in the State of Bahia, Brazil, where 99.3% of them were attacked by this spittlebug. Damage was more severe on branches and leaves. Tree death was, relatively, low while those cut showed 4.7 colonies of this spittlebug with 21.9 nymphs per colony. Cultural control with the removal of trees attacked was recommended.

Isolamento do alcalóide ricinina das folhas de Ricinus communis (Euphorbiaceae) através de cromatografias em contracorrente

Droplet counter-current chromatography, rotation locular counter-current chromatography and high-speed counter-current chromatography were applied to the preparative separation of the alkaloid ricinine from the dichloromethane extracts of Ricinus communis leaves. The solvent system used was composed of dichloromethane-methanol-water (93:35:72 v/v/v) and all techniques led to the isolation of large amounts of the alkaloid. The best result was obtained through HSCCC, since the ricinine yield was respectively 50% and 30% higher than when using RLCCC or DCCC.

Development and validation of a RP-HPLC method to determine the xanthyletin content in biodegradable polymeric nanoparticles

Xanthyletin is used as an inhibitor of the symbiotic fungus (Leucoagaricus gongylophorus) of the leaf-cutting ant (Atta sexdens rubropilosa), one of the most significant agricultural plague insects. The incorporation of this compound into nanoparticles is a promising approach to effectively control leaf-cutting ants. This study presents the development and validation of a specific analytical method using high-performance liquid chromatography (HPLC) for quantification of the xanthyletin content in biodegradable polymeric nanoparticles. The analytical methodology developed was specific, linear, accurate, precise, and robust. The absolute recovery of xanthyletin in colloidal suspensions was nearly 100%. The HPLC method proved reliable for the quantification of xanthyletin content in nanoparticle formulations.