Inhibition of the symbiotic fungus of leaf-cutting ants by coumarins

Leaf-cutting ants are known to be a serious pest for agriculture due to the high amounts of vegetal matter from crops used by them in order to cultivate a symbiotic fungus on which they rely for food and enzymes. The mutualism between the fungus and the ants is a point to be explored when alternative methods of control are being thought of. Considering that some plants are naturally resistant to phytophagous insects, some natural products (secondary metabolites) should be evaluated with respect to their insecticide and/or fungicide properties. In this paper we isolated eight coumarins from four different plant species and we determined their effect on the development of the symbiotic fungus of the leaf-cutting ant Atta sexdens. With the exception of clausarin, all the other coumarins were inhibitory from 64 μg mL-1 through 80 μg mL-1 and xanthyletin inhibited the fungus at 25 μ mL -1. ©2005 Sociedade Brasileira de Química.

Desenvolvimento e produtividade da cúrcuma (Curcuma longa L.) em função de doses de nitrogênio e potássio

The objective of the experiment was evaluating the development and production of turmeric, it was carried out at FCAV-UNESP, in Jaboticabal-SP, during 11/18/ 1999 to 7/18/2000, in randomized blocks, with three replications, factorial scheme 5 × 5. The factors evaluated were doses of N (0, 68, 136, 170 and 204 kg.ha-1) and doses of K2O (0, 92, 184, 230 and 276 kg.ha-1). The potassium doses influenced plant-mother's height significantly, answering lineally to the increment in the potassium fertilization. There was significant interaction between nitrogen and potassium on the number of leaves of the sons and total-plant. In the doses of 0 and 92 kg of K2O.ha-1 a lineal increase was observed in the number of leaves with the increment of the applied nitrogen. The potassium doses also influenced the foliar area of the sons and total-plant, reaching values of 3260 and 4971 cm2 per plant, respectively, for the application of 276 kg K2O.ha1. The number of sons had a lineal increment with the growing fertilization of potassium, contemplating on the dry mass of the aerial part of the sons. The production of rhizomes per plant wasn't influenced for the factors evaluated.

Toxicity of Cipadessa fruticosa to the leaf-cutting ants Atta sexdens rubropilosa (Hymenoptera: Formicidae) and their symbiotic fungus

Crude extracts from fruits, branches and leaves of Cipadessa fruticosa were assayed on Atta sexdens rubropilosa workers and their symbiotic fungus Leucoagaricus gongylophorus. The results showed that the majority of the extract tested reduced significantly (p < 0.05) the survival of the workers comparable to the neem oil. In addition, the dichloromethane extract from fruits and the hexane extract from branches of C. fruticosa inhibited in 80% the development of the symbiotic fungus. The hexane and dichloromethane extracts from fruits of C. fruticosa were fractionated and their fractions were subjected to new bioassays. Several fractions showed relevant activity on ants, but none of them inhibited significantly the fungal growth. The possibility of controlling leaf-cutting ants in the future using C. fruticosa is discussed.

Assessment of microfungi in fungus gardens free of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The purpose of this study was to determine the microfungi present in young nests of Atta sexdens rubropilosa when the fungus gardens were deprived of worker ants. The results were compared with another study in which worker ants had been killed by using toxic baits, and some species such as Acremonium kiliense, Escovopsis weberi, Moniliella suaveolens and Trichoderma sp. were confirmed among the most common inhabitants of this microenvironment, but differences in composition and proportion of species were observed. The importance and the role of these and other species of fungi within the symbiosis are discussed.

Leaf-cutting ants toxicity of limonexic acid and degraded limonoids from Raulinoa echinata. X-ray structure of epoxy-fraxinellone

Phytochemical survey of roots extracts of the South Brazilian endemic plant Raulinoa echinata Cowan, Rutaceae led to the isolation of known degraded limonoids: fraxinellone, fraxinellonone and epoxy-fraxinellone. The latter was previously isolated also from the stems, but the relative configuration could not be solved by NMR experiments. This paper deals with the X-ray diffraction analysis of epoxy-fraxinellone and its toxicity to leaf-cutting ants, describes the isolation of fraxinellonone and fraxinellone for the first time in Raulinoa. Epoxy-fraxinellone showed no toxicity to the leaf-cutting ants (Atta sexdens rubropilosa). The limonoid limonexic acid, isolated from stems, presented high toxicity to the leaf-cutting ants, diminishing considerably their longevity. ©2005 Sociedade Brasileira de Química.

Teores de nutrientes na folha e nos grãos de aveia-preta em função da adubação com fósforo e potássio

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The hypopharyngeal gland of leaf-cutting ants (Atta sexdens rubropilosa) (Hymenoptera: Formicidae)

The social behavior of ants is controlled by means of dynamic interactions between the environment and the individuals, mainly through pheromones. Among the exocrine glands is the hypopharyngeal gland (HG), located laterally over the pharyngeal plate. The present work aimed to describe and compare the HG in the different castes of the leaf-cutting ant Atta sexdens rubropilosa Forel 1908. The HG of different castes showed similar morphology varying only in relation to the secretory cell number. The secretory intracellular reservoir presented positive reaction to Xylidine Ponceau and P.A.S, indicating the presence of protein and polysaccharides in the secretion. Therefore, we suggest that the function of this gland would be the production of digestive enzymes and/or some sort of mucus, which, together with the secretions produced by the salivary glands of the thorax, would be related to the production of saliva.

Toxicity of Azadirachta indica to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The leaf-cutting ants are responsible for great economical damages in cultivated areas, and because of this characteristic, they have aroused the interest of researchers around the world. Thus, extracts of neem (Azadirachta indica, A. Juss) were tested through ingestion and contact experiments to investigate their toxicity to Atta sexdens rubropilosa workers. Crude extract of neem seed oil was toxic to the ants by ingestion in all concentrations tested but no toxicity was observed in contact experiments. However, crude extract of neem seed paste produced a significant toxicity to workers by ingestion and by contact.

Toxicity of straight-chain fatty acids to leaf-cutting ants Atta sexdens rubropilosa (Hymenoptera: Formicidae) and the symbiotic fungus Leucoagaricus gongylophorus

This work determined toxicity and attractiveness of straight-chain fatty acids (C 5 to C 12) to Atta sexdens rubropilosa (Forel) workers. The effect to the symbiotic fungus, Leucoagaricus gongylophorus (Singer) Möller, was also tested with the fatty acids C 6 to C 12. A strong mortality of leaf-cutting ants that were fed with an artificial diet containing fatty acids C to C at concentrations above 1.0 mg.ml -1 was observed. Rice flakes impregnated with solutions of these fatty acids were repellent to leaf-cutting ants. Contact experiments showed that treatments with C 6 and C 7 at concentration of 100 mg.ml -1 significantly reduced the survival rate of leaf-cutting ants. The fatty acids C 8 to C 11 were toxic to leaf-cutting ants when topically tested at concentration of 200 mg.ml -1. In relation to the fungus' bioassays, the fatty acids C 6 to C 12 at concentration of 0.1 mg.ml -1 inhibited 100% of the fungal development. Although when the concentration was reduced by half no inhibition effects were observed. The results showed that straight-chain fatty acids have desirable properties for controlling leaf-cutting ants since they directly interfere with both organisms of the symbiotic relationship. The potential of fatty acids as well as ways to control leaf-cutting ants with these compounds are discussed in this article.

The role of workers in transferring queen substances and the differences between worker castes in the leaf-cutting ant, Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae)

The present study focused on the relationship between the queen and workers in Acromyrmex subterraneus brunneus colonies mediated by the possible transfer of substances from the fertile to the sterile caste. The queens were submitted to different situations of physical limitation, i.e., they were kept isolated in cages with holes that only permitted the entry of workers but left the queen isolated. A tracer (water-soluble dye) was applied to the pronotum and gaster of the queen and its dispersal among workers was analyzed. The results demonstrated that the minor sub caste (0.7-0.9 mm) passed on the dye through allogrooming and self-grooming, or transferred the dye through trophallaxis to the major sub caste (1.2-2.0 mm) when they were not in direct contact with the queen. These findings indicate the communication and probable transfer of substances from the queen to the workers, as well as a substance transfer between workers.

Evaluation of antidiarrhoeal effects of Psidium guajava L. (Myrtaceae) aqueous leaf extract in mice

A crude aqueous extract of the leaves of the guava tree, Psidium guajava L. (Myrtaceae), were studied for antidiarrhoeal effects, to place one of its traditional medical uses. The extract induced a decrease in the propulsive movements of the intestinal contents in mice. These findings suggested that an aqueous extract of guava leaves may be used as an effective treatment for non-specific diarrhoea in folk medicine.

Dispersal of the delayed action insecticide sulfluramid in colonies of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Toxic baits are the most used control method for leaf-cutting ants due to their high effectiveness and because they are considered the safest for humans. Taking into account that the importance of leaf-cutting ants as pests, knowing the process by which dispersal and worker contamination is achieved becomes essential to understand several aspects about the functioning of a bait-borne AI (active ingredient) used in toxic baits. It has been established that an effective toxic bait should have a delayed- action AI, but its dispersion among the different sizes of workers is unknown. Workers of different sizes are involved in quite different tasks such foraging, cultivation of symbiotic macrofungus or control of deleterious microfungi. Therefore, we prepared a toxic bait containing the delayed-action AI sulfluramid and a dye (Rhodamine B) as an AI tracer in order to study dispersal and contamination in colonies, evaluated at different periods and in relation to different workers' sizes. Both field and laboratory colonies were evaluated. The great level of contamination, about 50% at 24 hours, in all sizes of workers demonstrates that worker contact with toxic bait is intense within this period. The distribution in field and laboratory colonies was similar. This contamination pattern is probably enough to cause the colony to die because of contamination of smaller workers, leading to the loss of control of the aggressive microfungi, which can quickly overgrow the symbiotic fungus culture. The dispersal dynamics of AI in leaf-cutting ant workers is important for investigations on the mode of action of this insecticide in the colony, and as a reference in future studies, such as those attempting to reduce the concentration of AIs in baits to reduce their environmental impact, or for facilitation of new AI screening.

Implementation of filters for image pre-processing for leaf analyses in plantations

In this work an image pre-processing module has been developed to extract quantitative information from plantation images with various degrees of infestation. Four filters comprise this module: the first one acts on smoothness of the image, the second one removes image background enhancing plants leaves, the third filter removes isolated dots not removed by the previous filter, and the fourth one is used to highlight leaves' edges. At first the filters were tested with MATLAB, for a quick visual feedback of the filters' behavior. Then the filters were implemented in the C programming language. At last, the module as been coded in VHDL for the implementation on a Stratix II family FPGA. Tests were run and the results are shown in this paper. © 2008 Springer-Verlag Berlin Heidelberg.

Soluções nutritivas no desenvolvimento do crisântemo cultivado em vaso

Although the information about nutritive solution for potted chrysanthemum is still incongruent, the objective of this work was to evaluate the effect of nutritive solutions in the development of Miramar chrysanthemum cultivated in pots at FCAV-UNESP, Jaboticabal-SP. The experiment had a randomized block design with split-plot evaluation and treatments corresponding to four nutritive solutions consisting of commercial fertilizers (S1, S2, S3, S4) evaluated at 0, 14, 28, 42, 56 and 70 days after rooting (DAE), with five replications. The plant height, leaf area, number of leaves, stem diameter, total dry matter mass of aerial part and roots; number and diameter of inflorescences, and physiological indexes of growth analysis were evaluated. The nutritive solutions did not interfere significantly in stem diameter, number and diameter of inflorescences and dry matter of roots, although S3 and S4 provided the highest plants, leaf area, number of leaves, total dry matter of aerial part. The physiological indexes showed that plants presented an accelerated growth from 14 to 28 DAE with S4 and S3 solutions and therefore they were considered the best solutions.

Proteína e qualidade de sementes de feijão (Phaseolus vulgaris L.) em função da adubação nitrogenada em plantio direto

Nitrogen limitation in the common bean (Phaseolus vulgaris L.) produces reduced productivity and seeds with low protein content and physiological quality. We assessed the effects of nitrogen fertilizer side dressing on the protein content and physiological quality of the seeds of P. vulgaris L. cultivars (cv.) IPR Juriti and Pérola grown in plots (soil type = Dystrophic Red Latosol) using a no-tillage system under a thick mulch of millet residues at the Experimental Station of UNESP-Ilha Solteira campus, located in Selvíria, MS (Köppen climate type = Aw) during autumn (March/June) and winter (June/September) 2005. For each cultivar, a randomized block experimental design was used with four replicates and factorially arranged treatments equivalent to 0, 30, 60, 90 and 120 kg.ha-1 added nitrogen (as urea, containing 45% of N) applied as a side dressing during the V4-3 and V4-6 phenological stage corresponding to the 3rd and 6th completely opened trifoliolate leaf on the main stem. Supplementation with up to 120 kg.ha-1 nitrogen promoted a greater increase in crude protein at V4-3 (unsupplemented = 17.6% and 16.3%; 120 kg.ha-1 N = 24.1% and 22.3% for cv. IPR Juriti and Pérola, respectively) than at V4-6 (unsupplemented = 19.2% and 18.3%; 120 kg.ha-1 N = 21.3% and 20.3% for cv. IPR Juriti and Pérola, respectively). About 90% of the crude protein from cv. IPR Juriti was composed of soluble protein compared to 72% for the cv. Pérola. Albumins and globulins represented about 80% of the soluble protein and prolamins were lower at 0.6%. In conclusion, nitrogen fertilization up to 120 kg.ha-1 applied as a side dressing at the V4-3 phenological stage in no-tillage under a thick mulch of millet promoted a greater increase of crude protein in common bean seeds than at the V4-6 stage. The highest accumulation of soluble protein occurred at 90 kg.ha-1 applied nitrogen without having a significant influence on the physiological quality of the seeds.