Purificação, caracterização e atividade bioinseticida de um inibidor de tripsina de sementes de Crotalaria pallida

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulphate fractionation, affinity chromatography on immobilized Trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named ITC, had Mr of 32.5 kDa by SDS-PAGE and was composed by two subunits with 27.7 and 5.6 kDa linked by disulphide bridges, a typical characteristic of Kunitz-Inhibitor family. ITC was stable until 50°C, and at 100°C its residual activity was of about 60%. Also, ITC was stable at pHs 2 to 12. The inhibition of trypsin by ITC was non-competitive, with a Ki of 8,8 x 10-7M. ITC inhibits weakly other serine proteinases such as chymotrypsin and elastase. The inhibition of papain (44% of inhibition), a cysteine proteinase was an indicative of the bi-functionality of ITC. In vitro assays against digestive proteinases from several Lepdoptera, Diptera and Coleoptera pests were made. ITC inhibited in 100% digestive enzymes of Ceratitis capitata (fruit fly), Spodoptera frugiperda and Alabama argillacea, the last one being a cotton pest. It also inhibited in 74.4% Callosobruchus maculatus (bean weevil) digestive enzymes, a Coleoptera pest. ITC, when added in artificial diet models, affected weakly the development of C. capitata larvae and it had a WD50 of 2.65% to C. maculatus larvae

Primer registro de Helcystogramma sp.(Lepidoptera: Gelechiidae) en Cuba.

Helcystogramma sp., es una nueva especie de microlepidóptero que afecta desde la siembra hasta la cosecha del boniato o camote (Ipomoea batatas L. Lam.) en Cuba. Debido a la falta de conocimientos y a la escasa bibliografía relacionada con las especies del género Helcystogramma en nuestro país, nos propusimos como objetivo conocer aspectos bio-ecológicos de este microlepidóptero, con la finalidad de proporcionar nuevos conocimientos para un futuro manejo de esta plaga. El trabajo se realizó desde septiembre 2013 a septiembre de 2014, en el Instituto de Investigaciones de Viandas Tropicales (INIVIT), Santo Domingo, Villa Clara, Cuba. Los insectos se estudiaron por observación directa en campos de I. batatas. La larva de este microlepidóptero es muy ágil, dobla la hoja, la cual pega con hilos de seda, creando una protección para su desarrollo hasta el estado de pupa, se alimenta dentro de esta cavidad a partir de la misma hoja, sin afectar la epidermis inferior. Sus poblaciones han aumentado considerablemente, llegando a 20 larvas/m2 en verano. La especies Helcystogramma sp., se cita por primera vez para Cuba y además se registra como una nueva especie de insecto que se alimenta del boniato. ABSTRACT. Primer registro de Helcystogramma sp. (Lepidoptera: Gelechiidae) en Cuba. Helcystogramma sp. is a new species of microlepidoptera affecting from planting to sweet potato (I. batatas L. Lam.) harvest crop in Cuba. Due to the lack of knowledgze and limited literature relate with the Helcystogramma genus in our country, the goal of this study was to know the bioecological aspects of such microlepidoptera, in order to provide new insights for future management of this pest. The study was conducted from September 2013 to September 2014 at the Research Institute of Tropical Root and Tuber Crops, Bananas and Plantains (INIVIT), Santo Domingo, Villa Clara, Cuba. Insects were studied by direct observation in sweet potato (I. batatas) fields. The microlepidoptera larvae is very agile, folds the sheet which sticks with silk threads, creating a protection for its development to the pupal stage. The larvae will feed in the cavity from the same leaf without affecting the lower epidermis. Their populations have increased significantly, reaching 20 larvae / m2 in summer. The species Helcystogramma sp., is cited for the first time in Cuba and it is reported as a new species of insect that feeds on sweet potato leaves.

Atividade bioinseticida e mecanismo de ação de vicilinas de sementes Erythrina velutina sobre moscas-das frutas Ceratitis capitata

The fruit fly Ceratitis capitata is considered the most destructive pest of the world fruitculture. Many pest management practices, mainly based on agrochemicals, have been developed to allow the world-wide commerce of fruit. Solutions to decrease the use of synthetic insecticides in agriculture are based on the development of new target-specific compounds which cause less damage to the environment, especially vegetal proteins with insecticidal effects. The aim of this work was to evaluate the deleterious effect of a purified vicilin of E. velutina (EvV) seeds to C. capitata larvae and adult insects and to investigate the mechanisms involved in these effects. EvV was purified, characterized and its deleterious effect was tested in bioassay systems. EvV mechanism of action was determined by immunodetection techniques and fluorescence localization in chitin structures that are present in C. capitata digestory system. EvV is a glycoprotein with affinity to chitin. Its molecular weight, of 216,57 kDa, was determined by gel filtration chromatography in FPLC system. Using SDS-PAGE, it was possible to observe EvV dissociation in two main subunits of 54,8 and 50,8 kDa. When it was submitted to eletrophoresis in native conditions, EvV presented only one band of acid characteristic. The WD50 and LD50 values found in the bioassays were 0,13% and 0,14% (w/w), respectively for the larvae. EvV deleterious effects were related to the binding to chitin structures presented in peritrophic membrane and gut epithelial cells, associated with its low digestibility in C. capitata digestive tract. The results described herein are the first demonstration of the larvicidal effects of plant protein on C. capitata larvae. EvV may be part of the pest management programs, in the toxic bait composition, or an alternative in plant improvement program

Boloceroides spp.

Boloceroides spp. are looking like an untidy mop, this anemone is sometimes seen in sea grass areas on many of our shores. It is possibly seasonal. Sometimes, large numbers are seen (up to 10-20 animals in a trip) and then none at all.  Tiny swimming anemones may sometimes be confused with Sea grass anemones which have translucent tentacles with tiny spots. The swimming anemone harbors symbiotic single-celled algae (zooxanthellae). The algae undergo photosynthesis to produce food from sunlight. The food produced is shared with the sea anemone, which in return provides the algae with shelter and minerals. The oral disk and tentacle muscles are used to obtain, retain, and ingest prey; in Boloceroides spp. tentacles can autotomize if it is needed to evade a predator. Tentacles can control body form by use of their endodermal muscles. Retractors are longitudinal muscles that will aid in withdrawing tentacles and the oral disk if they are exposed to the open air.  This hypothesis is furthered because in comparison to other sea anemones, Boloceroides is loosely attached to its respective substrate, thus allowing the pedal disk to detach quickly resulting in a rapid swimming response. Boloceroides can reproduce both sexually and asexually. As Anthozoans, Boloceroides produce sexually by bypassing the medusa life cycle stage; this allows Boloceroides (and all Anthozoans) to release their egg and sperm creating planula a bilaterally symmetrical, flattened, ciliated, motile larva.

Pempheris vanicolensis (Cuvier, 1831)

Pempheris vanicolensis has reported for the first time from Persian Gulf (Kish Island). Various aspects of the biology of P. vanicolensis Cuvier & Valenciennes, a recent Lessepsian (Suez Canal) immigrant into the Mediterranean, are given. Red Sea and Mediterranean populations were compared, and the results indicate that the spawning season is shortened in the Mediterranean, continuing from April to September, as opposed to year-round in the Red Sea. Descriptions of a ripe ovary and post-larva from the Mediterranean are given. Individuals reach a mean size of 10.8 cm in their first year, and 14.4 cm in the second year. Little change has occurred in the diet of the immigrant population, and both populations feed nocturnally, chiefly on larval and adult stages of planktonic crustaceans. Direct underwater observations on the diurnal behavior of the species show that the fish leave their daytime cave shelter at sunset, congregate at a nearby site and then migrate inshore and disperse into small groups to feed. Before dawn, they reassemble at the cave's entrance, and finally enter it at sunrise, after the school has built up. Observations on Persian Gulf sweepers show that the fish are segregated into size groups, ranging 15–18 cm adults at depths of 3 m.

Filling in gaps of Drosophila melanogaster urate degradation metabolic pathway using metabolomics approaches: towards the core metabolome of the fruit fly

The primary goal of systems biology is to integrate complex omics data, and data obtained from traditional experimental studies in order to provide a holistic understanding of organismal function. One way of achieving this aim is to generate genome-scale metabolic models (GEMs), which contain information on all metabolites, enzyme-coding genes, and biochemical reactions in a biological system. Drosophila melanogaster GEM has not been reconstructed to date. Constraint-free genome-wide metabolic model of the fruit fly has been reconstructed in our lab, identifying gaps, where no enzyme was identified and metabolites were either only produced or consume. The main focus of the work presented in this thesis was to develop a pipeline for efficient gap filling using metabolomics approaches combined with standard reverse genetics methods, using 5-hydroxyisourate hydrolase (5-HIUH) as an example. 5-HIUH plays a role in urate degradation pathway. Inability to degrade urate can lead to inborn errors of metabolism (IEMs) in humans, including hyperuricemia. Based on sequence analysis Drosophila CG30016 gene was hypothesised to encode 5- HIUH. CG30016 knockout flies were examined to identify Malpighian tubules phenotype, and shortened lifespan might reflect kidney disorders in hyperuricemia in humans. Moreover, LC-MS analysis of mutant tubules revealed that CG30016 is involved in purine metabolism, and specifically urate degradation pathway. However, the exact role of the gene has not been identified, and the complete method for gap filling has not been developed. Nevertheless, thanks to the work presented here, we are a step closer towards the development of a gap-filling pipeline in Drosophila melanogaster GEM. Importantly, the areas that require further optimisation were identified and are the focus of future research. Moreover, LC-MS analysis confirmed that tubules rather than the whole fly were more suitable for metabolomics analysis of purine metabolism. Previously, Dow/Davies lab has generated the most complete tissue-specific transcriptomic atlas for Drosophila – FlyAtlas.org, which provides data on gene expression across multiple tissues of adult fly and larva. FlyAtlas revealed that transcripts of many genes are enriched in specific Drosophila tissues, and that it is possible to deduce the functions of individual tissues within the fly. Based on FlyAtlas data, it has become clear that the fly (like other metazoan species) must be considered as a set of tissues, each 2 with its own distinct transcriptional and functional profile. Moreover, it revealed that for about 30% of the genome, reverse genetic methods (i.e. mutation in an unknown gene followed by observation of phenotype) are only useful if specific tissues are investigated. Based on the FlyAtlas findings, we aimed to build a primary tissue-specific metabolome of the fruit fly, in order to establish whether different Drosophila tissues have different metabolomes and if they correspond to tissue-specific transcriptome of the fruit fly (FlyAtlas.org). Different fly tissues have been dissected and their metabolome elucidated using LC-MS. The results confirmed that tissue metabolomes differ significantly from each other and from the whole fly, and that some of these differences can be correlated to the tissue function. The results illustrate the need to study individual tissues as well as the whole organism. It is clear that some metabolites that play an important role in a given tissue might not be detected in the whole fly sample because their abundance is much lower in comparison to other metabolites present in all tissues, which prevent the detection of the tissue-specific compound.

Análise estequiométrica da dinâmica não-linear de redes de reações químicas

Dissertação (mestrado)—Universidade de Brasília, Instituto de Química, Programa de Pós-Graduação em Química, 2016.

Tributos indiretos na cadeia produtiva do camarão

The state of Rio Grande do Norte counts with a relevant potential in the shrimp farming supply chain. In the larviculture step the state responds for more than half of the national production. In the farming step it is the second largest producer. In the industrial step, its industries have almost 40% of the shrimp processing capacity of the northeast of Brazil. However, this country has the highest tax rate comparing with the main shrimp producer countries. Considering the influence of taxes in the competition among companies, the main goal of this research is to analyze the impact of indirect taxes in the above steps of the supply chain. To achieve it, it will be used the data of the 2011 Census of the Shrimp Farming and it will be applied the Herfindahl-Hirschman Index to identify the market form of those steps. In order to contribute with the characterization of the supply chain, CEO´s of farms and industries will be interviewed. The price-elasticity of the shrimp larvae, the in natura shrimp and the processed shrimp will be analyzed in order to verify the possibility that each one of those three steps has to pass-through the onus of the end of benefit over the ICMS. The data analysis shows that the larviculture step functions as a duopoly and, facing the end of that benefit, it will be able to pass-through most its onus to the farming step. On the other hand, this step functions similar to a perfect competing market, which diminishes its capacity to pass-through that onus to the processing step. This step operates as oligopoly with a lower concentration than the larviculture step but, due to the fact that it faces an oligopsony, it will end up assuming most of that onus, which will cause a decrease in the amount of processed shrimp. It is concluded that the end of that benefit would impact negatively, in this state, the supply chain at all, but mainly the farming and the industrial steps

Aspectos biologicos do acaro vermelho do tomateiro.

Alguns aspectos biologicos de Tetranychus evansi Baker & Pritchard foram estudados em laboratorio, incluindo a duracao das formas jovens e a capacidade reprodutiva, em plantas de tomate var. Rossol. As duracoes dos estagios do ovo, larva, protoninfa e deutoninfa foram de 4,8; 2,6, 2,4 e 3,2 dias, respectivamente, correspondendo a uma duracao total de, aproximadamente treze dias de ovo a adulto, para ambos os secos. Os niveis mais altos de oviposicao foram obtidos do quarto ao decimo ia, aproximadamente. Uma media de 105,7 ovos foram postos durante todo o periodo de oviposicao. Provavelmente, a utilizacao de folhas medianas ou basais, ao inves de apicais, como substrato teria resultado em um ciclo biologico mais curto e em uma oviposicao total maior do que os obtidos neste estudo.

Large national series of patients with Xq28 duplication involving MECP2: Delineation of brain MRI abnormalities in 30 affected patients.

Xq28 duplications encompassing MECP2 have been described in male patients with a severe neurodevelopmental disorder associated with hypotonia and spasticity, severe learning disability, stereotyped movements, and recurrent pulmonary infections. We report on standardized brain magnetic resonance imaging (MRI) data of 30 affected patients carrying an Xq28 duplication involving MECP2 of various sizes (228 kb to 11.7 Mb). The aim of this study was to seek recurrent malformations and attempt to determine whether variations in imaging features could be explained by differences in the size of the duplications. We showed that 93% of patients had brain MRI abnormalities such as corpus callosum abnormalities (n = 20), reduced volume of the white matter (WM) (n = 12), ventricular dilatation (n = 9), abnormal increased hyperintensities on T2-weighted images involving posterior periventricular WM (n = 6), and vermis hypoplasia (n = 5). The occipitofrontal circumference varied considerably between >+2SD in five patients and <-2SD in four patients. Among the nine patients with dilatation of the lateral ventricles, six had a duplication involving L1CAM. The only patient harboring bilateral posterior subependymal nodular heterotopia also carried an FLNA gene duplication. We could not demonstrate a correlation between periventricular WM hyperintensities/delayed myelination and duplication of the IKBKG gene. We thus conclude that patients with an Xq28 duplication involving MECP2 share some similar but non-specific brain abnormalities. These imaging features, therefore, could not constitute a diagnostic clue. The genotype-phenotype correlation failed to demonstrate a relationship between the presence of nodular heterotopia, ventricular dilatation, WM abnormalities, and the presence of FLNA, L1CAM, or IKBKG, respectively, in the duplicated segment.

The transcriptome of metamorphosing flatfish

Background Flatfish metamorphosis denotes the extraordinary transformation of a symmetric pelagic larva into an asymmetric benthic juvenile. Metamorphosis in vertebrates is driven by thyroid hormones (THs), but how they orchestrate the cellular, morphological and functional modifications associated with maturation to juvenile/adult states in flatfish is an enigma. Since THs act via thyroid receptors that are ligand activated transcription factors, we hypothesized that the maturation of tissues during metamorphosis should be preceded by significant modifications in the transcriptome. Targeting the unique metamorphosis of flatfish and taking advantage of the large size of Atlantic halibut (Hippoglossus hippoglossus) larvae, we determined the molecular basis of TH action using RNA sequencing. Results De novo assembly of sequences for larval head, skin and gastrointestinal tract (GI-tract) yielded 90,676, 65,530 and 38,426 contigs, respectively. More than 57 % of the assembled sequences were successfully annotated using a multi-step Blast approach. A unique set of biological processes and candidate genes were identified specifically associated with changes in morphology and function of the head, skin and GI-tract. Transcriptome dynamics during metamorphosis were mapped with SOLiD sequencing of whole larvae and revealed greater than 8,000 differentially expressed (DE) genes significantly (p < 0.05) up- or down-regulated in comparison with the juvenile stage. Candidate transcripts quantified by SOLiD and qPCR analysis were significantly (r = 0.843; p < 0.05) correlated. The majority (98 %) of DE genes during metamorphosis were not TH-responsive. TH-responsive transcripts clustered into 6 groups based on their expression pattern during metamorphosis and the majority of the 145 DE TH-responsive genes were down-regulated. Conclusions A transcriptome resource has been generated for metamorphosing Atlantic halibut and over 8,000 DE transcripts per stage were identified. Unique sets of biological processes and candidate genes were associated with changes in the head, skin and GI-tract during metamorphosis. A small proportion of DE transcripts were TH-responsive, suggesting that they trigger gene networks, signalling cascades and transcription factors, leading to the overt changes in tissue occurring during metamorphosis.

Efeito da privação alimentar sobre alguns componentes bioquímicos (proteína, ADN e ARN) e o comportamento alimentar em larvas de pregado (Scophthalmus maximus,L.) em condições de cultivo

Dissertação de mest. em Estudos Marinhos e Costeiros, especialização em Aquacultura, Unidade de Ciências e Tecnologias dos Recursos Aquáticos, Univ. do Algarve, 1994

BRS ARAGUAIA: híbrido de melão amarelo, produtivo e saboroso.

Equipe técnica: Valter Rodrigues Oliveira, Leonardo Silva Boiteux, Agnaldo Donizete Ferreira de Carvalho, Ailton Reis, André Nepomuceno Dusi, Carlos Alberto Lopes, Celso Luiz Moretti, Geni Livtin Villas Bôas, Ítalo Moraes Rocha Guedes, Jadir Borges Pinheiro, João Maria Charchar, Leonora Mansur Mattos, Mirtes Freitas de Lima, Ronessa Bartolomeu de Souza, Werito Fernandes de Melo, Gláucia Salles Cortopassi Buso, Marco Antônio Ferreira, Fernando Antonio de Souza Aragão, Francisco das Chagas Vidal Neto, Waldelice Oliveira de Paiva, João Ribeiro Crisóstomo, Ricardo Elesbão Alves, José Luiz Mosca, Heloisa Almeida Cunha Figueiras, Antônio Apoliano dos Santos, Nivaldo Duardo Costa (CPATSA), Joston Simão de Assis (CPATSA), Rita de Cássia Souza Dias (CPATSA), José Amauri Buso, Marcos Coelho.

Tomate industrial no submedio Sao Francisco e as pragas que limitam sua producao.

Neste trabalho sao relatados: a importancia economica do tomateiro, a melhor epoca de seu plantio, e as principais pragas que limitam a producao do tomate industrial na regiao do submedio Sao Francisco. As pragas sao: a traca-do-tomateiro, Scrobipalpula absoluta; o microacaro ou acaro-do-bronzeamento, Aculops lycopersici; o acaro-vermelho, Tetranychus evansi; a broca-dos-frutos, Heliothis zea e Pseudoplusia oo; a larva-minadora, Liromyza sativae e a lagarta-rosca Agrotis ipsilon.

Resistência de variedades de Vigna unguiculata ao ataque de Lyriomiza sativae (Diptera, Agromyzidae).

Estudou-se a resistência de 50 variedades de Vigna unguiculataWalp ao ataque de Liriomyza sativae Blanchard, sob condições de irrigação por sulco. As variedades que sofreram os menores e os maiores danos causados pelo inseto foram: VITA-3 e 5F-PI-112, respectivamente, observando-se susceptibilidades intermediárias das variedades estudadas entre estes dois extremos.