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.

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.

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.

Caracterização morfoanatômica de plântulas de Gomphrena elegans Mart. (Amaranthaceae)

Aiming to describe seedlings' morphology and cotyledons and eophylls anatomy of Gomphrena elegans Mart., 100 seeds were put into plastic bags, with sand and fertilized soil (1:3), kept in the greenhouse under room conditions, with 50% shady and watered daily. Cotyledonous petiole and limb and seedlings eophylls were sectioned in median and border. They were then analyzed in transversal and paradermic sections. Seedlings initial morphology is epigeal-foliar, the root is axial, and hypocotyls and epicotyl are reddish. Venation is of pinnate type, camptodrome and brochidrodome in cotyledons and eophylls. Petiole and leaf show uniseriated epidermis with cells of varied sizes and anomocytic stomata. Druses of calcium oxalate occurred in leaves and petiole. Dorsiventral mesophyll shows one palisade parenchyma layer and several lacunary ones.

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.

Leaf spray fertilization of boron and zinc on productivity and fruit quality of Japanese pear tree

The cultivation of fruit plants from temperate climate in tropical or subtropical regions can be a good income alternative for the producer. However, due to the little existent information about cultivation of those fruit plants, the producers use imported techniques of other producing areas, or even an association of practices used for other fruit plants, pointing out the leaf spray fertilization of micronutrients without appropriate scientific base. In this context, the objective of this study was to verify the effect of the leaf spray fertilization of B and Zn on productivity and fruit quality of Japanese pear tree. The experiment was conducted from 2004 to 2005, in Ilha Solteira, in northwestern São Paulo State-Brazil. The climate is, according to the Köpppen Classification, tropical wet and dry (Aw). The 'Okusankichi' cultivar, grafted on Pyrus communis L. rootstock was used as well as doses of 110 g.ha-1 of B and 250 g.ha-1 of Zn in each application. The treatments were: T1. water, T2. boric acid, T3. zinc sulfate, T4. T2 + T3, T5. boric acid + urea + citric acid + EDTA, T6. zinc sulfate + urea + citric acid + EDTA, T7. T5 + T6, T8. boric acid + urea + citric acid + EDTA + sodium molibdate + sulfur + calcium chloride, T9. zinc sulfate + urea + citric acid + EDTA + Fe sulfate + Mn sulfate + Mg sulfate and, T10. T8+T9. A randomized blocks design was used and the averages were compared by Tukey test. In the first crop the mixture of boric acid with quelating agents were efficient to supply B to the plants and zinc sulfate plus quelating agents were efficient to increase Zn leaf content. However, the productivity and the fruit quality were not influenced by the leaf spray of B and Zn. In the second crop the leaf content of B and Zn and the productivity were not influenced by the leaf spray; the boric acid and the zinc sulfate with or without quelating agents increased the contents of total soluble solids and, the boric acid with or without quelating agents increased the contents of total titratable acidity.

SEM studies on the leaf indumentum of six Melastomataceae species from Brazilian Cerrado

The wide diversity of their trichomes, which vary from simple unicellular to very complex structures, is a remarkable characteristic in Melastomataceae. This paper characterizes the leaf indumentum of Miconia albicans (Sw.) Triana, M. chamissois Naudin, M. fallax DC., M. ligustroides (DC.) Naudin, Microlepis oleaefolia (DC.) Triana and Rhynchanthera dichotoma DC., typical species from Brazilian cerrado. Samples collected from the median third of young and mature leaf blades were processed following the usual scanning electron microscopy techniques (SEM). We observed ten morphological types of trichomes and four of emergences. With five different types, four of which are reported for the first time, Rhynchanthera dichotoma is the species that presents the most diverse indumenta. A mixed type of trichome formed by a glandular and a branched non-glandular portion called lateral-gland was observed in M. ligustroides. Such non-glandular portion presents different degrees of development. A correlation is suggested between the stage of development of the non-glandular portion and the exposition to light of these lateral gland on young leaves.

Measuring and analyzing color and texture information in anatomical leaf cross sections: An approach using computer vision to aid plant species identification

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)