RESUMO

Cysteine peptidases are involved in physiological processes of insect development and have been considered as potential targets for the development of insect control strategies. In this study, we obtained a recombinant cysteine cathepsin L (AsCathL) from leaf-cutting ant (Atta sexdens), a species from the order Hymenoptera who causes enormous damage to crops, natural forests and reforested areas. RT-qPCR showed AsCathL expression throughout insect development and in all body parts of the adult insect analysed, suggesting its role as a lysosomal cathepsin. AsCathL encodes a protein of 320 amino acid residues consisting of a pro-peptide and the mature with amino acids sequence over 67% similarity with lysosomal cathepsin L of species from Lepidoptera and Diptera. Phylogenetic tree revealed that AsCathL is very similar to predicted cathepsins found in other ants. Recombinant AsCathL was expressed in insoluble form by Escherichia coli Arctic Express (DE3) RIL, purified under denaturing conditions and refolded. The enzyme showed hydrolytic activity in vitro towards synthetic substrate Z-Phe-Arg-AMC at acidic pH. Synthetic inhibitor E-64 acted against peptidase activity and a study regarding the interaction between E-64 and AsCathL using nuclear magnetic resonance (NMR) revealed that 83.18% from all E-64 molecules are irreversibly bound to AsCathL. In addition, the proteolytic activity of AsCathL was strongly inhibited by recombinant sugarcane cystatins with Ki ranging from 0.6 nM to 2.95 nM. To the best of our knowledge this is the first report characterizing a cysteine peptidase from leaf-cutting ants, which may contribute to future studies of ants' cathepsins.

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RESUMO

Eibesfeldtphora tonhascai Brown (Diptera: Phoridae) is a natural enemy of Atta sexdens Linnaeus and Atta laevigata Smith and is considered a promising candidate for the biological control of these ants. The aim of this study was to verify if E. tonhascai has a preference for specific sizes of A. sexdens foragers and whether this parasitoid demonstrated host species preference when comparing parasitism of A. sexdens and A. laevigata. Worker ants were classified into four different size groups and introduced into an attack arena with one E. tonhascai female. To verify host species preference, both A. sexdens and A. laevigata workers were placed in the attack arena together with one E. tonhascai female. A no-choice test was also performed, when each ant species was offered to a phorid female at different times. Eibesfeldtphora tonhascai performed the highest number of inspection flights, attempted attacks, and actual attacks against A. sexdens workers with a cephalic capsule size of 3-5 mm. When comparing attacks on A. sexdens and A. laevigata workers, they were equally inspected, attacked, and the total parasitism rates were similar. However, the phorid adult emergence rate was significantly higher in the A. sexdens parasitized workers. We conclude that E. tonhascai females express preference for parasitizing larger hosts (cephalic capsule width of 3-5 mm), but do not display a preference for host species when comparing A. sexdens versus A. laevigata, and thus can similarly contribute to both leafcutter ants' natural biological control.

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RESUMO

Strategies for the control of leaf-cutting ants have mainly involved granular baits based on fipronil and sulfluramid as active ingredients, which are commonly coated with attractive citrus-based substances. Their constant use and the lack of alternatives in the market may favor the perception of these substances by ants, causing rejection for foraging and consequent difficulty in their control. In this respect, this study examined the mortality of leaf-cutting ants of the genera Atta and Acromyrmex subjected to direct application with dry powders, in laboratory conditions. As a preliminary treatment, a commercial antiseptic talc powder (C. A. P. T.) was used, followed by isolated treatments that corresponded to its components with potential insecticidal action, namely, salicylic acid, sulfur, boric acid, zinc oxide, in addition to an inert talc powder (Quimidrol®) as the control. For each treatment, 40 (worker) ants, whose activity was reduced due to remaining in a refrigerator prior to the treatment, were placed in transparent crystal polystyrene ('Gerbox' type) and were sprinkled with a salt shaker. The ants were kept at 25 ± 2 °C, under a 12-h photophase, and cumulative mortality was recorded every 24 h, considering dead ants as those that were unable to maintain the natural position of their body, i.e., even dying ants were considered dead when they exhibited no reaction when touched by a paintbrush. When sprinkled on ants, C. A. P. T. causes 40% mortality in both species 24 h after application. Subsequent studies of the components of this commercial product have found that when sprinkled on both Atta sexdens rubropilosa and Acromyrmex crassispinus, salicylic acid causes 100% mortality of workers in the first 24 h. This result provides a new prospect of control through a low environmental-impact product, representing an alternative for control in nests in the field that can contribute to the integrated control of ants.(AU)

As estratégias de controle de formigas-cortadeiras têm sido realizadas principalmente com iscas granuladas, à base dos princípios ativos fipronil e sulfluramida, que são comumente revestidas por substâncias atrativas cítricas. O uso constante das mesmas e a falta de alternativas no mercado podem favorecer a percepção de tais substâncias pelas formigas, causando rejeição para forrageamento e consequente dificuldade do seu controle. Neste sentido, o objetivo do trabalho foi avaliar a mortalidade de formigas-cortadeiras dos gêneros Atta e Acromyrmex, submetidas a aplicação direta com pós secos, em laboratório. Como tratamento preliminar, utilizou-se: um talco antisséptico comercial e, posteriormente seus componentes, com potencial ação inseticida, isoladamente, sendo: ácido salicílico, enxofre, ácido bórico, óxido de zinco, além de um talco inerte (Quimidrol®), como testemunha. Para cada tratamento foram utilizadas 40 formigas (operárias) acondicionadas em poliestireno cristal transparente (Tipo Gerbox), que foram polvilhadas com auxílio de um saleiro, estando com atividade reduzida, devido a permanência em geladeira, previamente ao tratamento. As formigas foram mantidas em 25 ± 2°C, fotofase de 12 horas e a mortalidade acumulada foi observada a cada 24 horas, considerando-se formigas mortas aquelas que não conseguiam manter a posição natural do corpo, ou seja, mesmo formigas moribundas foram consideradas mortas, quando não exibiam reação, ao serem tocadas por um pincel. O talco antisséptico comercial, quando polvilhado sobre as formigas, causa mortalidade de 40%, em ambas as espécies, 24 horas após as aplicações. Estudos subsequentes dos componentes do referido produto comercial detectaram que o ácido salicílico, quando polvilhado tanto para Atta sexdens rubropilosa quanto para Acromyrmex crassispinus, causa mortalidade de 100% das operárias nas primeiras 24 horas. Esse resultado abre uma nova perspectiva de controle, com produto de baixo impacto ambiental, sinalizando uma alternativa de controle em ninhos a campo, podendo contribuir para o controle integrado de formigas.(AU)

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RESUMO

Leaf-cutter ants perform a series of specialized behaviors in preparing plant substrates for their symbiotic fungus. This process may be related to contamination of workers by substances such as insecticides, leading us to hypothesize that substances are spread among workers through behaviors they perform to grow the fungus. To test this hypothesis, we analyzed the behavioral acts of workers during the processing of the pellets by using a fat-soluble tracing dye, since the active ingredient that composes toxic baits, used for control of leaf-cutter ants, is fat-soluble. The frequencies of performed behaviors were recorded and the number of dyed workers was assessed after fungus cultivation. The most frequent behavior is allogrooming and corresponds to 45.87% of the contamination process in workers, followed by holding, licking, and cutting pellets, which account for 40.22% of the process. After pellet processing, the workers had their external and internal morphological structures marked by the tracing dye-93.75% and 79.25%, respectively. These results confirm that behaviors performed during fungus cultivation contribute to dispersing substances such as insecticides, causing the contamination of workers.

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RESUMO

Flower and leaf herbivory might cause relevant and negative impacts on plant fitness. While flower removal or damage by florivores produces direct negative effects on plant fitness, folivores affect plant fitness by reducing resource allocation to reproduction. In this study, we examine the effects of both flower and leaf herbivory by leaf-cutting ants on the reproductive success of the shrub species Miconia nervosa (Smith) Triana (Family Melastomataceae) in a fragment of Atlantic Forest in Northeast Brazil. We conducted a randomized block-designed field experiment with nine replicates (blocks), in which three plants per block were assigned to one of the three following treatments: undamaged plants (ant exclusion), leaf-damaged plants (ant exclusion from reproductive organs, but not from leaves), and flower + leaf-damaged plants (no exclusion of ants). We then measured flower production, fruit set, and fruit production. Our results showed that flower + leaf-damaged plants reduced flower production nearly twofold in relation to undamaged plants, while flower set in leaf-damaged plants remained constant. The number of flowers that turned into fruits (i.e., fruit set), however, increased by 15% in flower + leaf-damaged plants, while it slightly decreased in leaf-damaged compared to undamaged plants. Contrastingly, fruit production was similar between all treatments. Taken together, our results suggest a prominent role of ant floral herbivory across different stages of the reproductive cycle in M. nervosa, with no consequences on final fruit production. The tolerance of M. nervosa to leaf-cutting ant herbivory might explain its high abundance in human-modified landscapes where leaf-cutting ants are hyper-abundant.

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RESUMO

The aim of this study was to determine whether worker self-grooming, allogrooming, and direct contact promotes the dispersal of substances among members of the colony. For this purpose, a tracer (Sudan III dye) was applied topically to a worker ant and the social interactions between the worker with the tracer and workers without the tracer were studied. Additionally, the worker heads were dissected to visualize whether or not the post-pharyngeal gland was stained. The post-pharyngeal glands from 50% to 70% of workers were stained depending on the size of the group. With the increase in the experimental group size, the frequency of interactions between workers increased, with touching being the most frequent behavior. The tracer dye was probably passed on by direct contact between workers, followed by self-grooming and allogrooming. These behaviors are responsible for the rapid dispersal of substances among colony members as observed in our experiment. The results therefore support the hypothesis that contact with substances promotes the contamination of nestmates, even in the absence of feeding, serving as a model for further studies on the contamination of workers with the active ingredients of insecticides.

RESUMO

ABSTRACT In leafcutter ants the division of labour is associated to worker size variation clustered in four subcastes. In this work we used Atta sexdens Forel (1908) as a model to test the hypothesis that each subcaste expresses its own chemical signature comprised of cuticular lipids. To assess it, we extracted epicuticular compounds by using nonpolar solvent hexane and analysed the samples in a combined Gas Chromatography-Mass Spectrometer (GC-MS). We found 24 hydrocarbons with carbon chains ranging from 19 to 39 atoms most of them classified as linear and branched alkanes. No compound occurred in the cuticle of specific worker subcaste, however, the relative proportion pattern varied greatly among them. Our results suggest that although subcastes have similar chemical signatures, significant differences in their relative proportions may play an important role between nestmate and group identification.

RESUMO

Cuticular hydrocarbons (CHCs) are traditionally considered to be one of the most important chemical cues used in the nestmate recognition process of social hymenopterans. However, it has been suggested that in the leafcutter ant genus Atta, it is not the CHCs, but the alarm pheromone that is involved in the nestmate recognition process. In this study we used a laboratory population of Atta sexdens to explore the association between their CHC profile variation and intraspecific aggression. In the first part of the experiment, four colonies were divided into two groups with distinct diets to stimulate differentiation of their CHC profiles. In the second part of the experiment, all colonies received the same diet to examine resemblance of chemical profiles. At the end of each part of the experiment we extracted the CHCs from workers. The results demonstrated that colonies that shared the same food resource had similar cuticular hydrocarbon profiles. Furthermore, colonies were significantly more aggressive towards conspecifics that used a different foliar substrate and consequently had greater differences in their cuticular chemical composition. This study suggests that the CHC profiles of A. sexdens can be affected by the foliar substrates used, and that the CHCs are used in the nestmate recognition process of this species.

RESUMO

The objective of this study was to characterize phenolic compounds and evaluate the effect, under laboratory conditions, of the cassava leaf powder methanol extract on the development of fall armyworm Spodoptera frugiperda and of leaf-cutting ant Atta sexdens rubropilosa. The extract was incorporated into an artificial diet, to which the armyworm was exposed, at concentrations of 250, 500, 1,000 and 1,500 mg kg-1, in order to evaluate biological characteristics. Soon after the insects emergence, another experiment was conducted to verify the possible sub lethal activity of the extract; therefore, S. frugiperda couples were isolated in cages and eggs were collected and counted. The Manihot esculenta Crantz extract caused a reduction in the percentage of armyworm survival, as well as in the eggs number. Then, the same extract was solubilized in 10% ethanol and applied to ants; mortality was observed, compared to the control. It is possible to conclude that the M. esculenta Crantz leaf powder methanolic extract, containing gallic acid and catechin, is a promising alternative to control S. frugiperda and Atta sexdens rubropilosa.

Objetivou-se com o presente trabalho verificar o efeito do extrato metanólico da farinha das folhas de mandioca contra Spodoptera frugiperda e Atta sexdens rubropilosa. O referido extrato, em diferentes concentrações, foi incorporado na dieta, a qual foi exposta a lagarta para avaliar características biológicas. Logo após a emergência dos insetos, casais de S. frugiperda foram isolados em gaiolas para verificar a possível atividade sub-letal do extrato e as posturas foram recolhidas para a quantificação. O extrato de Manihot esculenta Crantz provocou redução na porcentagem de sobrevivência das lagartas, bem como no número de ovos.  A seguir o mesmo extrato foi solubilizado em etanol 10 % e aplicado em formigas, observou-se a mortalidade em comparação com a testemunha. Os experimentos foram conduzidos em delineamento inteiramente casualizados. Os resultados obtidos demonstraram que o extrato metanólico da farinha das folhas de mandioca apresenta-se como uma alternativa promissora ao controle de S. frugiperda e de Atta sexdens rubropilosa, necessitando no entanto de novos estudos para isolamento e identificação das substâncias bioativas.

RESUMO

The objective of this study was to characterize phenolic compounds and evaluate the effect, under laboratory conditions, of the cassava leaf powder methanol extract on the development of fall armyworm Spodoptera frugiperda and of leaf-cutting ant Atta sexdens rubropilosa. The extract was incorporated into an artificial diet, to which the armyworm was exposed, at concentrations of 250, 500, 1,000 and 1,500 mg kg-1, in order to evaluate biological characteristics. Soon after the insects emergence, another experiment was conducted to verify the possible sub lethal activity of the extract; therefore, S. frugiperda couples were isolated in cages and eggs were collected and counted. The Manihot esculenta Crantz extract caused a reduction in the percentage of armyworm survival, as well as in the eggs number. Then, the same extract was solubilized in 10% ethanol and applied to ants; mortality was observed, compared to the control. It is possible to conclude that the M. esculenta Crantz leaf powder methanolic extract, containing gallic acid and catechin, is a promising alternative to control S. frugiperda and Atta sexdens rubropilosa. 

Objetivou-se com o presente trabalho verificar o efeito do extrato metanólico da farinha das folhas de mandioca contra Spodoptera frugiperda e Atta sexdens rubropilosa. O referido extrato, em diferentes concentrações, foi incorporado na dieta, a qual foi exposta a lagarta para avaliar características biológicas. Logo após a emergência dos insetos, casais de S. frugiperda foram isolados em gaiolas para verificar a possível atividade sub-letal do extrato e as posturas foram recolhidas para a quantificação. O extrato de Manihot esculenta Crantz provocou redução na porcentagem de sobrevivência das lagartas, bem como no número de ovos.  A seguir o mesmo extrato foi solubilizado em etanol 10 % e aplicado em formigas, observou-se a mortalidade em comparação com a testemunha. Os experimentos foram conduzidos em delineamento inteiramente casualizados. Os resultados obtidos demonstraram que o extrato metanólico da farinha das folhas de mandioca apresenta-se como uma alternativa promissora ao controle de S. frugiperda e de Atta sexdens rubropilosa, necessitando no entanto de novos estudos para isolamento e identificação das substâncias bioativas.

RESUMO

In vitro culture of the mutualistic fungus of leaf-cutting ants is troublesome due to its low growth rate, which leads to storage problems and contaminants accumulation. This paper aims at comparing the radial growth rate of the mutualistic fungus of Atta sexdens rubropilosa Forel in two different culture media (Pagnocca B and MEA LP). Although total MEA LP radial growth was greater all along the bioassay, no significant difference was detected between growth efficiencies of the two media. Previous evidences of low growth rate for this fungus were confirmed. Since these data cannot point greater efficiency of one culture medium over the other, MEA LP medium is indicated for in vitro studies with this mutualistic fungus due its simpler composition and translucent color, making the analysis easier.

Assuntos

RESUMO

In vitro culture of the mutualistic fungus of leaf-cutting ants is troublesome due to its low growth rate, which leads to storage problems and contaminants accumulation. This paper aims at comparing the radial growth rate of the mutualistic fungus of Atta sexdens rubropilosa Forel in two different culture media (Pagnocca B and MEA LP). Although total MEA LP radial growth was greater all along the bioassay, no significant difference was detected between growth efficiencies of the two media. Previous evidences of low growth rate for this fungus were confirmed. Since these data cannot point greater efficiency of one culture medium over the other, MEA LP medium is indicated for in vitro studies with this mutualistic fungus due its simpler composition and translucent color, making the analysis easier.

RESUMO

In vitro culture of the mutualistic fungus of leaf-cutting ants is troublesome due to its low growth rate, which leads to storage problems and contaminants accumulation. This paper aims at comparing the radial growth rate of the mutualistic fungus of Atta sexdens rubropilosa Forel in two different culture media (Pagnocca B and MEA LP). Although total MEA LP radial growth was greater all along the bioassay, no significant difference was detected between growth efficiencies of the two media. Previous evidences of low growth rate for this fungus were confirmed. Since these data cannot point greater efficiency of one culture medium over the other, MEA LP medium is indicated for in vitro studies with this mutualistic fungus due its simpler composition and translucent color, making the analysis easier.

RESUMO

This research was developed in hibrid urograndis (Eucalyptus urophylla Eucalyptus grandis)plantation belonging to Jari Celulose S.A., in Pará State, Brazil, from August 1994 to January 1995. The objective was to test one granulated bait with sulfluramid and three others with chlorpirifos in three dosages compared to 10 grams of dodecachlor (0.45%) bait per square meter of ant nest against Atta sexdens sexdens(Hymenoptera: Formicidae). The bait with sulfluramid showed 77.78%; 100.00%; and 88.90% efficiency against A. sexdens sexdensat 6, 8, and 10 grams while the dodecachlor bait showed 100.00% control. The other baits and dosages showed efficiency below 45.00%.

Este trabalho foi realizado em plantios de eucalipto urograndis (Eucalyptus urophylla Eucalyptus grandis)da Jari Celulose S.A., no município de Almeirim, Pará, no período de agosto/94 a janeiro/95. Foi testada a eficiência de uma isca granulada com sulfluramida e de três outras com clorpirifós, em três dosagens cada por metro quadrado de formigueiro, no controle de Atta sexdens sexdens(Hymenoptera: Formicidae). Os resultados foram comparados com os de uma isca à base de dodecacloro (0,45%) a 10 gramas por metro quadrado de formigueiro. A isca granulada com sulfluramida a 6, 8 e 10 gramas por metro quadrado de formigueiro apresentou 77,78%, 100,00% e 88,90% de controle, respectivamente, assemelhando-se à isca a base de dodecacloro que apresentou 100,00% de formigueiros mortos. As outras iscas, em qualquer dosagem, apresentaram eficiência abaixo de 45,00%.

RESUMO

This research was developed in hibrid urograndis (Eucalyptus urophylla Eucalyptus grandis)plantation belonging to Jari Celulose S.A., in Pará State, Brazil, from August 1994 to January 1995. The objective was to test one granulated bait with sulfluramid and three others with chlorpirifos in three dosages compared to 10 grams of dodecachlor (0.45%) bait per square meter of ant nest against Atta sexdens sexdens(Hymenoptera: Formicidae). The bait with sulfluramid showed 77.78%; 100.00%; and 88.90% efficiency against A. sexdens sexdensat 6, 8, and 10 grams while the dodecachlor bait showed 100.00% control. The other baits and dosages showed efficiency below 45.00%.

Este trabalho foi realizado em plantios de eucalipto urograndis (Eucalyptus urophylla Eucalyptus grandis)da Jari Celulose S.A., no município de Almeirim, Pará, no período de agosto/94 a janeiro/95. Foi testada a eficiência de uma isca granulada com sulfluramida e de três outras com clorpirifós, em três dosagens cada por metro quadrado de formigueiro, no controle de Atta sexdens sexdens(Hymenoptera: Formicidae). Os resultados foram comparados com os de uma isca à base de dodecacloro (0,45%) a 10 gramas por metro quadrado de formigueiro. A isca granulada com sulfluramida a 6, 8 e 10 gramas por metro quadrado de formigueiro apresentou 77,78%, 100,00% e 88,90% de controle, respectivamente, assemelhando-se à isca a base de dodecacloro que apresentou 100,00% de formigueiros mortos. As outras iscas, em qualquer dosagem, apresentaram eficiência abaixo de 45,00%.