The pulvinus endodermal cells and their relation to leaf movement in legumes of the Brazilian cerrado

Legume pulvini have a clearly delimited endodermis, whose variable content has been associated with the velocity and type of leaf movement: pulvini in leaves with fast nastic movement contain starch grains; pulvini in leaves with slow nastic movements have calcium oxalate crystals as well as starch grains in the endodermis. However, the studies carried out to date have involved few legume species. This study therefore purported to examine the consistency of this hypothesis in other legumes. Thus, the structure and content of the pulvinus endodermal cells of nine legumes of the Brazilian cerrado, with different types and velocities of leaf movement, were investigated: slow nyctinastic and heliotropic movements (Bauhinia rufa, Copaifera langsdorffii, Senna rugosa - Caesalpinioideae; Andira humilis and Dalbergia miscolobium - Faboideae; Stryphnodendron polyphyllum - Mimosoideae), slow heliotropic movement (Zornia diphylla - Faboideae), and fast seismonastic and slow nyctinastic and heliotropic movements (Mimosa rixosa and Mimosa flexuosa - Mimosoideae). Samples were prepared following standard plant anatomy and ultrastructure techniques. The endodermis of all the species contains starch grains. In the species displaying only slow movements, calcium oxalate prismatic crystals were observed in addition to starch grains, except in Zornia diphylla. in conclusion, oxalate crystals occur only in endodermal cells of pulvini that display slow movements, while starch grains are always present in pulvinus endodermal cells of plants with any kind of movement.

Colonization of rocky and leaf pack substrates by benthic macroinvertebrates in a stream in southeast Brazil.

A comparative analysis was conducted of the colonization by benthic macroinvertebrates of rocky and leaf pack substrates, both natural and artificial. This colonization was evaluated by season, with the objective of ascertaining the influence of rainfall on the rate of colonization. The total density of macroinvertebrates after 21 days of colonization was significantly greater in the dry than in the wet season. When the substrate types were compared, artificial leaf pack substrate presented the smallest density for both seasons. In the wet season, Chironomidae, Leptohyphidae, Hydropsychidae, Elmidae, immature stages of Trichoptera, and Hydroptilidae showed a more representative density. In the dry season, Chironomidae, Baetidae and Oligochaeta were the most abundant taxa. The artificial rocky substrate used in this experiment was the most appropriate, due to its resemblance with natural substrate conditions in terms of the maintenance of the structural integrity of the substrate throughout the experimental period. Successional and seasonal effects were of great relevance, playing an important role in the colonization process.

Behavior of Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers during the preparation of the leaf substrate for symbiont fungus culture

The behavioral repertory of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) workers marked by size category was studied during the preparation of the leaf substrate in the laboratory. The workers were marked according to three physical castes, i.e., minima, generalist and forager. Seven types of behavioral acts were recorded for each caste, with the following frequencies: licking leaf fragments (64.6%), holding fragments on the surface of the fungus garden (16.4%), shredding the fragments (6.0%), chewing and crimping the edges of the fragments (9.0%), incorporating the fragments (2.7%), touching the surface of the fungus with their mandibles and other mouthparts after incorporation (0.3%), and depositing fecal fluid (0.1%). The minima workers were found to be more specialized in the activities related to the preparation of the leaf substrate, which represented 52% of the total number of tasks performed. The generalists performed 40.3% of these tasks, and the foragers 7.9%. Licking the substrate was the behavior most frequently recorded and performed for a longer period of time. In this way, the workers may feed and at the same time eliminate microorganisms that are harmful to the symbiont fungus. The smaller castes, minima and generalists, are those most responsible for the preparation of the leaf substrate and predominate within a colony. From a practical viewpoint, with the introduction of toxic bait containing insecticides, for example, these size categories will be those most intensely intoxicated, especially through the behavior of licking bait pellets. On the basis of the data obtained about these behaviors, we may raise the hypothesis that trophallaxis in not the major factor triggering contamination with an insecticide among the workers of a colony.

The effect of plant diversity on fungus garden development and foraging behavior of leaf-cutting ants (Hymenoptera: Formicidae)

The leaf-cutting ants forage a wide variety of plant species, used for symbiotic fungus cultivation. To better understand this tripartite complex interaction, 24 colonies of Acromyrmex subterraneus brunneus were conditioned for 4 months to 6 different plants (Citrus spp., Ligustrum spp., Acalypha spp., Eucalyptus spp., Alchornea triplinervia, Melia spp.), to verify the influence of conditioning on foraging behavior of workers. The effect of plants on symbiotic fungus development was studied separately, through macerated plants in Agar and culture medium A as the control. During foraging, workers presented polyphagic foraging behavior, refusing the plants to which they were conditioned. The selection of plants is not correlated with the plant substrate that promotes good development of symbiotic fungus. Such results demonstrate the importance of plant diversity for fungus garden maintenance.

Toxicity of sesame extracts to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Extracts of different sesame plant (Sesamum indicum, Linnaeus) organs were tested through ingestion and contact experiments to investigate their toxicity to Atta sexdens rubropilosa (Forel) workers. Dichloromethane extracts of seeds were toxic to the leaf-cutting ants and the factor responsible for the toxicity does not show seasonal occurrence.

Toxicity of sesame seed to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Extracts of the ripe seeds of the sesame plant (Sesamum indicum, Linnaeus) were tested through contact experiments to investigate their toxicity to Atta sexdens rubropilosa workers. Dichloromethane extract of seeds was toxic to the ants and the factor responsible for this effect was distributed through the ethyl acetate fraction. This fraction was divided into four sub fractions composed of: A) triglycerides, B) monoglycerides + diglycerides + triglycerides, C) diglycerides + sesamoline + sesamine and D) sesamine. However, when these sub fractions were separated, no toxicity was observed. Therefore, in order to determine why the activity was lost, the concentration of each sub fraction was duplicated, and the possible combinations among them were also tested. We concluded that the toxicity to the ants is due mainly to a mixture of triglycerides, and sesamoline or the combination of sesamoline + sesamine can be a synergistic factor in this fraction.

Astilbin toxicity to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The toxicity of astilbin, isolated from Dimorphandra mollis, was tested in laboratory ingestion bioassays for the leaf-cutting ant Atta sexdens rubropilosa Forel. Worker ants that were fed an artificial diet daily to which astilbin was added had a higher mortality rate than the controls. The substance astilbin was incorporated into the diet with and without the utilization of a solvent. Results for both methodologies utilized were similar and indicate that astilbin reduces the median survival of ants. All concentrations tested showed toxic effects against ant workers, and the statistical comparison of survivorship rates from control and treated groups was significantly different, confirming the insecticidal properties of the substance astilbin.

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.

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.

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.

Antiulcer effects of Ethyl acetate fraction of Carpolobia lutea leaf

Carpolobia lutea leaves (CLL) (Polygalaceae) were earlier screened and the antiulcer ethnomedicinal claim established. This article seeks to quantitatively isolate, elucidate the active compounds from most active CLL fraction. Fractionation was by semi-preparative HPLC; the active fraction was subjected to radical scavenging assays (RSA) and quantification of the total phenolic content (TPC) were also executed. Results: Ethyl acetate fraction (EAF) was observed to be the most pharmacologically active antiulcer fraction when screened using acute ulcer models induced in rodents. The EAF demonstrated significant (p < 0.05-0.001) antiulcer activity in various in vivo induced ulceration models by reducing the ulcer index and increasing the preventive ratio. The EAF demonstrated > 70% in TPC and < 20 % in RSA. Cinnamic and coumaric acids derivatives were isolated from EAF. Cinnamic acids have been implicated and patented as antiulcer agent. Isolated compounds could in part mediate the observed pharmacological activities which lend credence to its ethnobotanical uses.