ABSTRACT. The effect of sub-lethal doses of Azadirachta indica (Meliaceae) oil on the midgut of Spodoptera frugiperda (Lepidoptera, Noctuidae). The fall armyworm, Spodoptera frugiperda, is one of the major field pests for maize production. It is mainly controlled by means of synthetic, and more recently by resistant cultivar of maize expressing Bt toxins. The neem tree, Azadirachta indica, is a plant that can potentially control insects with the advantage of being food and environmental safe. The aim of this study was to assess the effect of neem oil on the development and survival of S. frugiperda caterpillars by assessing histological alterations caused on their midgut. Newly hatched caterpillars were submitted to three neem oil concentrations: 0.006; 0.05; 0.4%, which were added to their artificial diet. Ten 3 rd instar caterpillars, taken from each treatment, were submitted to histological analysis. The alimentary canals from the specimens were fixed in Baker for 12 hours, desiccated and diaphanized in alcohol/xylol (1:1) and xylol. After placing the samples in paraffin, they were sliced in 8 µm sections and stained with hematoxylineosin stain. The neem oil added to the diet of S. frugiperda caused total mortality at dose of 0.4% whilst still in the first instars, prolonged the larval and pupal stages, and reduced the pupal weight. Histo-physiological alterations such as degeneration of the epithelial lining of the midgut and in the peritrophic matrix were found at all concentrations of neem oil.KEYWORDS. Fall armyworm; histological analysis; insecticidal plants; neem. RESUMO. Efeito de doses subletais do óleo de Azadirachta indica (Meliaceae) no mesêntero de Spodoptera frugiperda (J. E. Smith) (Lepidoptera, Noctuidae). A lagarta-do-cartucho, Spodoptera frugiperda, é a mais importante praga da cultura do milho. Esta é usualmente, controlada por inseticidas sintéticos e mais atualmente por meio de variedades resistentes de milho com a toxina Bt. O nim Azadirachta indica é planta com potencial no controle de insetos, que possui as vantagens de segurança alimentar e ambiental. Objetivou-se com este estudo avaliar o efeito do óleo do nim no desenvolvimento e sobrevivência de lagartas de S. frugiperda em decorrência das alterações no mesêntero. As lagartas recém nascidas foram submetidas a três concentrações do óleo de nim: 0,006; 0,05; 0,4%, adicionadas à dieta artificial. Dez lagartas do 3º instar, retiradas de cada tratamento, foram submetidas à análise histológica. O canal alimentar dos indivíduos foi fixado em Baker por 12 horas, desidratado, diafanizado em álcool/xilol (1:1) e xilol. Após a inclusão em parafina, as amostras foram seccionadas em 8 µm e coradas pela técnica de hematoxilina-eosina. Observou-se que o óleo de A. indica adicionado à dieta de S. frugiperda causa: mortalidade total na dosagem 0,4% nos primeiros instares, aumento na duração do período larval e pupal, redução no peso de pupas. Alterações histofisiológicas, como degeneração do epitélio do revestimento do mesêntero, da matriz peritrófi...

Venoms are a rich source for the discovery of molecules with biotechnological applications, but their analysis is challenging even for state-of-the-art proteomics. Here we report on a large-scale proteomic assessment of the venom of Loxosceles intermedia, the so-called brown spider. Venom was extracted from 200 spiders and fractioned into two aliquots relative to a 10 kDa cutoff mass. Each of these was further fractioned and digested with trypsin (4 h), trypsin (18 h), pepsin (18 h), and chymotrypsin (18 h), then analyzed by MudPIT on an LTQ-Orbitrap XL ETD mass spectrometer fragmenting precursors by CID, HCD, and ETD. Aliquots of undigested samples were also analyzed. Our experimental design allowed us to apply spectral networks, thus enabling us to obtain meta-contig assemblies, and consequently de novo sequencing of practically complete proteins, culminating in a deep proteome assessment of the venom. Data are available via ProteomeXchange, with identifier PXD005523.

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This is the first study on the hemolymph from a spider of the Loxosceles genus. These animals are responsible for a great number of envenomation cases worldwide. Several studies on Loxosceles venoms have been published, and the knowledge about the venom and its toxins is considerable, not only regarding the biological and biochemical characterization, but also regarding structural, genetic and phylogenetic approaches. However, the literature on Loxosceles hemolymph is nonexistent. The main goal of the present study was to characterize biochemically the hemolymph content, and especially, to identify its different hemocytes. Moreover, many papers have already shown molecules whose source is the hemolymph and their very interesting activities and biomedical applications, for example, antifungal and antibacterial activities. A 2D-SDS-PAGE of brown spider hemolymph showed approximately 111 spots for pH 3-10 and 150 spots for pH 4-7. A lectin-blotting assay showed that hemolymph carbohydrate residues were similar to those found in venom. Several types of TAG and DAG phospholipids were found in the hemolymph and characterized by HPTLC and mass spectrometry. Four different hemocytes were characterized in Loxosceles intermedia hemolymph: prohemocyte, plasmatocyte, granulocyte and adipohemocyte. This paper opens new possibilities on toxinology, studying an unknown biological material, and it characterizes a source of molecules with putative biotechnological applications.

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