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.

Method for the evaluation of insecticidal activity over time in Atta sexdens rubropilosa workers (Hymenoptera: Formicidae)

The active ingredients used in the formulation of toxic baits for leaf-cutting ants (Hymenoptera: Formicidae) should possess a delayed action defined as an insecticidal activity whereby worker mortality is ≤15% at 24 hours and ≥90% at 21 days. Serious shortcomings have occurred in the search for new active ingredients, such as the initial selection of fenoxycarb, copper oxychloride and diflubenzuron, compounds considered very promising but whose inefficiency was verified only later, indicating methodological problems. In view of this situation, we developed a classification method for insecticidal activity over time using workers of the leaf-cutting ant Atta sexdens rubropilosa Forel. The insecticides used were fipronil, sulfluramid GX071HB and sulfluramid GX439, vehicled in an attractive pasty formulation prepared based on citrus pulp. The results obtained were consistent from a toxicological viewpoint and agreed with the literature in terms of the control of colonies. Sulfluramids were found to possess a delayed action at a broad range of concentrations, in agreement with the fact that these substances are highly effective in the control of all leaf-cutting ant species. The smaller range of concentrations of fipronil with delayed action is probably related to its lower efficacy for species more difficult to control such as Atta capiguara (Forti et al. 2003). We discuss the importance of relating behavioral particularities to the specific feeding habits of leaf-cutting ants, with methodological adequacy of the assessment of insecticides aimed at toxic baits.

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.

Laboratory assessment of two active ingredients for control of coptotemes gestroi (Isoptera: Rhinotermitidae)

The subterranean termite Coptotermes gestroi Wasmann (Isoptera: Rhinotermitidae) is a pest of major economic importance in urban environments of southeastern Brazil. Compared to using pesticides for termite control, termite bait products target termites more specifically and also address environmental contamination issues. In this point of view, we performed two different bioassays in order to evaluate the efficacy of boric acid and fipronil against different populations of C. gestroi. The results showed that concentrations between 2000 and 3000 ppm of boric acid caused approximately 100 percent mortality in termites. Concentrations between 0.01 and 0.0001 ppm of fipronil resulted in 100% termite mortality after 2 wk exposure. The data displayed a fast mortality of termites contaminated with fipronil, even with small concentrations, and therefore it is not a suitable product to be used in baits against C. gestroi. The present study showed a delayed toxicity of boric acid against the subterranean termite C. gestroi which suggests a need for further field tests.

Toxicological and histopathological effects of boric acid on atta sexdens rubropilosa (Hymenoptera: Formicidae) workers

The current study compared the toxicity of different concentrations of boric acid in adult workers of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae), with toxicological bioassays, and examining the dose-dependent and time-dependent histopathological changes, of the midgut, Malpighian tubules, and postpharyngeal glands. Our results revealed the importance of conducting toxicological bioassays combined with morphological analyses of the organs of ants chronically exposed to insecticides used in commercial ant baits. In vitro bioassays showed that boric acid significantly decreases the survivorship of workers regardless of concentration, whereas the morphological data suggested progressive dose-dependent and time-dependent changes in the organs examined, which were evident in the midgut. The midgut is the first organ to be affected, followed by the postpharyngeal gland and Malpighian tubules. This sequence is in agreement with the absorption pathway of this chemical compound in the midgut, its transference to the hemolymph, possibly reaching the postpharyngeal glands, and excretion by the Malpighian tubules. These progressive changes might be due to the cumulative and delayed effect of boric acid. Our findings provide important information for the understanding of the action of boric acid in ant baits in direct and indirect target organs. © 2010 Entomological Society of America.

First record of chilodonella hexasticha (ciliophora: Chilodonellidae) in brazilian cultured fish: A morphological and pathological assessment

Chilodonelids are small ciliated protozoans found worldwide and can be dangerous in culture conditions. This study presents morphometric data on the ciliate Chilodonella that is found in cultured Nile tilapia (Oreochromis niloticus), native bait fish tuvira (Gymnotus aff. inaequilabiatus) and native pacu (Piaractus mesopotamicus) and includes a histopathological assessment of the changes that occur in the pacu. For parasitic diagnosis, skin and gill samples were scraped onto slides, dried at room temperature, stained with Giemsa or impregnated with silver nitrate, and the measurements were obtained from photomicrographs. In the diseased pacu, the first gill arch was collected and fixed in a 10% buffered formalin solution for histopathological analysis. Parasite specimens from the different collection sites were identified morphologically as C. hexasticha Kiernik (1909). Diseased fish exhibited depigmentation, skin ulceration, scale loss, excessive mucus production and gill lesions. Histopathological analysis of pacu gills displayed epithelial proliferation with mononuclear inflammatory infiltrate, hemorrhages, and scattering necrosis. In Brazilian-farmed fish this is the first record of C. hexasticha, which has great pathogenic potential in cultured freshwater species. In addition, two new hosts are presented. © 2012 Elsevier B.V.

Population dynamics of the freshwater crab Dilocarcinus pagei in tropical waters in southeastern Brazil

In the northwestern region of the state of São Paulo, Brazil, the freshwater crab Dilocarcinus pagei Stimpson, 1861 is usually sold as bait for several fish species for sport fishing. Thus, knowledge of the reproductive biology of this species is essential to provide information for maintaining natural stocks and implementing future farming initiatives. The objective of this study was to investigate the population dynamics of the red freshwater crab D. pagei, focusing on reproductive and juvenile recruitment periods, sex ratio, and individual growth. Sampling was performed monthly from October 2005 to September 2007 in the Marimbondo-Fumas Hydroelectric Plant reservoir, Rio Grande, municipality of Icém in northwestern São Paulo state. The individuals were sexed, carapace widths were measured, and the gonads were examined macroscopically. In total, 1002 individuals were analyzed, consisting of 568 males and 434 females, of which 4 were ovigerous and 35 carried early juveniles in the incubatory chamber. The largest numbers of reproductive individuals of both sexes were recorded during the winter and spring months before spawning, which occurred in the spring. Intense juvenile recruitment and the highest rainfall levels were observed during the Southern Hemisphere summer. The results indicate that spawning can be considered seasonal, and the simultaneous emergence of juveniles with high rainfall levels is probably the result of the higher availability of food and potential sites for protection against predators. The skewed sex ratio (♂ 1:0.76 ♀) may be associated with growth rates, mortality, food restriction, behavioral differences, migration, and differential habitat use between the sexes. The similar maximum size (CW∞) and growth constant (k) for males (k = 1.78 y-1, CW = 61.43 mm) and females (k = 1.67 y-1, CW = 60.66 mm) may be associated with different energy investment strategies between the sexes. Additionally, a closed season is proposed for the species. © The Crustacean Society, 2013. Published by Brill NV, Leiden.

The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication

Background: Yellow fever virus (YFV) belongs to the Flavivirus genus and causes an important disease. An alarming resurgence of viral circulation and the expansion of YFV-endemic zones have been detected in Africa and South America in recent years. NS5 is a viral protein that contains methyltransferase and RNA-dependent RNA polymerase (RdRp) domains, which are essential for viral replication, and the interactions between NS5 and cellular proteins have been studied to better understand viral replication. The aim of this study was to characterize the interaction of the NS5 protein with eukaryotic translation initiation factor 3 subunit L (eIF3L) and to evaluate the role of eIF3L in yellow fever replication. Methods. To identify interactions of YFV NS5 with cellular proteins, we performed a two-hybrid screen using the YFV NS5 RdRp domain as bait with a human cDNA library, and RNApol deletion mutants were generated and analyzed using the two-hybrid system for mapping the interactions. The RNApol region involved was segmented into three fragments and analyzed using an eIF3L-expressing yeast strain. To map the NS5 residues that are critical for the interactions, we performed site-direct mutagenesis in segment 3 of the interaction domain (ID) and confirmed the interaction using in vitro assays and in vivo coimmunoprecipitation. The significance of eIF3L for YFV replication was investigated using eIF3L overexpression and RNA interference. Results: In this work, we describe and characterize the interaction of NS5 with the translation factor eIF3L. The interaction between NS5 and eIF3L was confirmed using in vitro binding and in vivo coimmunoprecipitation assays. This interaction occurs at a region (the interaction domain of the RNApol domain) that is conserved in several flaviviruses and that is, therefore, likely to be relevant to the genus. eIF3L overexpression and plaque reduction assays showed a slight effect on YFV replication, indicating that the interaction of eIF3L with YFV NS5 may play a role in YFV replication. Conclusions: Although the precise function of eIF3L on interactions with viral proteins is not entirely understood, these results indicate an interaction of eIF3L with YF NS5 and that eIF3L overexpression facilitates translation, which has potential implications for virus replication. © 2013 Morais et al.; licensee BioMed Central Ltd.

Análise da produtividade e viabilidade econômica de quatro sistemas de berçários do camarão-da-Amazônia, Macrobrachium amazonicum

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desenvolvimento gonadal inicial e reversão sexual em Astyanax altiparanae (Teleostei, characidae)

Pós-graduação em Ciências Biológicas (Biologia Celular e Molecular) - IBRC

Cultivo multitrófico e multiespacial do camarão-da-amazônia e tilápia-do-nilo, com e sem o uso de substratos

Pós-graduação em Aquicultura - FCAV

Influência da polpa cítrica, do óleo e de fungos filamentosos na atratividade de iscas tóxicas à Atta sexdens rubropilosa (Hymenoptera,Formicidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Determinação do território de forrageamento e avaliação do uso de micro porta-iscas para as saúvas Atta sexdens rubropilosa Forel, 1908 e Atta laevigata Fr. Smith, 1858 (Hymenoptera, Formicidae)

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Toxicidade de formicidas utilizados em pastagens sobre a formiga não-alvo Ectatomma brunneum (Hymenoptera, Formicidae, Ectatomminae) e seus efeitos na dinâmica populacional em condições de laboratório

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Seleção de substratos celulósicos atrativos para o cupim Coptotermes gestroi (Isoptera: Rhinotermitidae)

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