A produção brasileira de alimentos cresce em ritmo vertiginoso, havendo previsão de expansão nos próximos anos. Nota-se, porém, que o modelo atual empregado ao controle de doenças e pragas agrícolas, tem causado diversos problemas de ordem ambiental, social e econômica. Uma alternativa à redução de tais impactos, tem sido a implementação do controle biológico no modelo de manejo. Neste trabalho, avaliou-se o controle biológico do fungo fitopatogênico Botrytis cinerea por compostos produzidos pela linhagem ASBV-1T. Os screenings \"in vitro\" demonstraram que a linhagem produz quitinases e metabólitos secundários ativos, conhecidamente descritos como fatores importantes ao controle de fitopatógenos. A caracterização parcial do complexo enzimático, indicou que as quitinases produzidas pela linhagem ASBV-1T apresentam maior atividade em meio alcalino (pH 6.8-10.0), temperatura de 30°C e possuem estimativa de peso molecular superior a 100 kDa. Os ensaios \"in vivo\", realizados em morangos comerciais (cv. Oso Grande) demonstraram a efetividade dos subprodutos bioativos de caráter ionóforos (monactina, dinactina, trinactina, tetranactina e valinomicina) em controlar a infestação de B. cinerea, durante a fase de pós-colheita, em condições de armazenamento. Por meio da suplementação do meio de cultivo por sais inorgânicos, a via biossintética responsável pela expressão dos compostos de interesse pode ser ativada. A adição de MgS04 resultou em um aumento de 400% na produção de macrotetralídeos e 20% de valinomicina. Contudo, a expressão dos macrotetralídeos foi completamente inibida pela adição de ZnSO4. dobrando a produção de valinomicina. De acordo com os estudos de taxonomia polifasica, o isolado ASBV-1T, apresenta caracteristicas quimiotaxonômicos e moleculares pertinentes ao gênero Streptomyces, formando uma nova linha filética, no subclado das espécies S. albolongus, S. cavourensis subsp. cavourensis e S. celluloflavus. Diante do supra citado, propõem-se que o isolado seja reconhecido como a linhagem tipo de Streptomyces araujoniae sp. nov. Os resultados obtidos neste trabalho, permitem afirmar que a linhagem S. araujoniae sp. nov. apresenta potencial de ação contra o fitopatógeno B. cinerea, podendo auxiliar no controle desta doença durante a fase de pós-colheita da cultura de morangos. Ainda resta, para estudos futuros, o desenvolvimento de formulação específica para o emprego destes compostos no manejo da cultura, bem como ensaios toxicológicos, assegurando a viabilidade de aplicação de tais compostos. / The Brazilian food production has been growing fast in recent years and is expected to expand even more. However, the models used for the control of pests and diseases in agriculture has been widely questioned because they cause problems of environmental, social and economic. To reducing the use of chemical agents, has been the implementation of biological control to the management model to reduce this problematic. In this study, we evaluated the biological control of fungal pathogen Botrytis cinera of active compounds produced by strain ASBV-1T. In vitro assays indicated the presence of chitinase and antifungal metabolites in compounds produced by strain ASBV-1T and may been contributed to the antagonistic activity againts the fungal pathogenic. Partial characterization of the enzymatic complex showed the highest production of chitinase under alkalines conditions at pH (6.8-10.0), 30°C and has estimated molecular weight above 100 kDa. The test in vivo, preformed in commercial strawberries (cv. Oso Grande) demonstrated the effectiveness of bioactive products of character ionophores (monoactin, dinactin, trinactin, tetranactin and valinomycin) controlling the infestation of B. cinerea during the post-harvest, in store conditions. The biosynthetic pathway responsible for the expression of the compounds of interest can be activated by supplementations of the culture medium by inorganic salts mainly with MgSO4 resulted in increse of 400% in the production of macrotetralides and 20% valinomycin. However, the expression of the macrotetralides was inhibited when added ZnSO4, bending the production of valinomycin. According to polyphasic taxonomic studies, the isolated ASBV-1T has chemotaxonomic and molecular characteristics of the genus Streptomyces, which formed a new phyletic line, in subcalde of the S. albolongus, S. cavourensis subsp. cavourensis e S. celluloflavus species. Then propose that the isolate is recognized as the type strain of Streptomyces araujoniae sp. nov. The results obtained in this study revealed that S. araujoniae sp. nov. has action potential against the pathogen B. cinerea, can help control this disease during the post-harvest strawberries. Remains for future studies, the development of specific formulation of the employment of such compounds to crop management and toxicological tests, ensuring the feasibility of using such compounds.

Hematopoietic stem cells facilitate lifelong production of a diverse repertoire of functional mature blood cells. They are a critical biological reservoir that enable organisms to endure physiological challenges such as inflammation, disease, and age. The functional maintenance of hematopoietic stem cells depends not only on intrinsic cell pathways, but also on extrinsic cues that guide core behaviors like homing and self-renewal. Careful study of these extrinsic regulatory networks can deepen our appreciation of fundamental stem cell biology and motivate therapeutic approaches to treat hematologic disease. Here I show how derangement of the bone marrow regulatory environment perturbs normal hematopoiesis, and demonstrate the dependence of hematopoietic stem cells on a circulating endocrine factor.

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Orientador: Carlos Frederico Wilcken / Banca: Carlos Gilberto Raetano / Banca: Edivaldo Domingues Velini / Banca: Geraldo Papa / Banca: Robinson Osipe / Resumo: Os herbicidas são capazes de promover alterações fisiológicas nas plantas cultivadas não suscetíveis a eles, podendo acarretar mudanças na bioecologia dos insetos que utilizam tais plantas para sobrevivência, inclusive insetos-praga e seus inimigos naturais. Além disso, aplicações seqüenciais ou associadas de herbicidas e inseticidas podem influenciar a eficácia de controle dos insetos-praga. O objetivo deste estudo foi avaliar o impacto de herbicidas aplicados em pós-emergência em plantas de milho sobre o desenvolvimento de Spodoptera frugiperda (Lepidoptera: Noctuidae) e sobre seus inimigos naturais em campo e laboratório. A interação entre herbicidas e inseticidas sobre o controle da praga também foi avaliada. Em laboratório, plantas de milho foram pulverizadas com os herbicidas bentazona, atrazina, nicossulfuron e 2,4-D (doses recomendadas) e folhas fornecidas a lagartas neonatas de S. frugiperda, para o acompanhamento do desenvolvimento do inseto. De forma semelhante, pupas de S. frugiperda obtidas das lagartas criadas sobre as folhas de milho tratadas com os herbicidas foram oferecidas a fêmeas do parasitóide de pupas Trichospilus diatraeae (Hymenoptera: Eulophidae) para acompanhamento do desenvolvimento e taxa de parasitismo do parasitóide. Em campo os herbicidas foram aplicados nas plantas de milho e a população de S. frugiperda foi monitorada semanalmente. Quinze dias após a aplicação dos herbicidas larvas da praga foram coletadas e transferidas individualmente para potes contendo dieta artificial para avaliação do parasitismo natural. O efeito dos herbicidas sobre o controle químico de S. frugiperda foi realizado da seguinte forma: larvas do inseto foram alimentadas com dieta artificial contendo herbicidas e após 48 h foram colocadas para se alimentar... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Herbicides can promote physiological changes in non susceptible crops and may lead to changes in bioecology of insects that use these plants for their survival, especially pests and their natural enemies. In addition, herbicides applications next or in mixture with insecticides can affect the effectiveness of pest control. The aim of this study was to evaluate the impact of post-emergence herbicides applied in corn plants on the development of Spodoptera frugiperda (Lepidoptera: Noctuidae) and their natural enemies in field and laboratory. The interaction between herbicides and insecticides on pest control was also evaluated. In laboratory corn plants were sprayed with the herbicides bentazon, atrazine, nicosulfuron and 2,4-D and leaves were supplied to neonate larvae of S. frugiperda for development evaluation. Similarly, S. frugiperda pupae obtained from caterpillars reared on treated leaves with herbicides were offered to females of the Trichospilus diatraeae (Hymenoptera: Eulophidae). The development and parasitism rate by parasitoid was evaluated. In field conditions the herbicides were applied in corn plants and the population of S. frugiperda was monitored weekly. Fourteen days after the herbicides application pest larvae were collected and transferred individually to pots containing artificial diet to evaluate the natural parasitism. The effect of herbicides on the chemical control of S. frugiperda was performed as follows: the insect larvae were fed on artificial diet containing herbicides 48 h before exposition to insecticides. After larvae were placed to feed on corn leaves treated with synthetic insecticides chlorpyrifos, deltamethrin, methomyl, etofenprox and spinosad, these same insecticides... (Complete abstract click electronic access below) / Doutor

Helicoverpa armigera é uma espécie que até 2013 era considerada praga quarentenária A1 no Brasil. Tratamentos com inseticidas são indispensáveis para o manejo de H. armigera em quase todas as culturas do mundo. Uma alternativa à utilização de inseticidas sintéticos para o controle de pragas é o uso de parasitoides do gênero Trichogramma. Em consideração a grande quantidade de aplicações de agroquímicos no sistema de produção de grãos, torna-se essencial avaliar a seletividade dos inseticidas utilizados, garantindo informações que auxilie na manutenção do T. pretiosum na área, podendo potencializar o controle biológico de H. armigera, sendo isto, importante para o estabelecimento do manejo integrado de pragas nas grandes culturas. Portanto, objetivou-se com esse trabalho: estudar a toxicidade aguda e a persistência de inseticidas sobre o parasitoide; estudar o efeito de inseticidas sobre a fase pupal do parasitoide; determinar a interferência dos inseticidas na capacidade de parasitismo. Dos inseticidas testados em condições laboratoriais, Nomolt®, Avatar®, Belt®, Premio® e Intrepid® foram considerados inócuos para adultos. Lorsban® e Engeo Pleno® são nocivos à fase de pupa, enquanto Acefato®, Pirate® e Belt® apesar de inócuos afetam as gerações seguintes. Pirate®, Lorsban® e Engeo Pleno interferem na capacidade de parasitismo, e Acefato® tem efeito deletério na geração que tem contato com o resíduo. Os inseticidas Nomolt®, Avatar®, Belt®, Acefato®, Premio® e Intrepid® foram classificados como inseticidas de vida curta (classe 1). Lorsban® tem efeito moderadamente persistente (classe 3) e Engeo Pleno® e Pirate® classificados como persistentes (classe 4). Portanto, visando um bom programa de MIP, deve-se optar pelo uso de inseticidas seletivos, ou inseticidas nocivos para adultos do parasitoide podem ser utilizados no campo desde que estes não estejam ocorrendo naturalmente ou que não coincida com épocas de liberação do mesmo. / Helicoverpa armigera is a specie that until 2013 was considered A1 quarantine pest in Brazil. Treatments with insecticides are indispensable for the management of H. armigera in almost every crops in the world. An alternative to the use of synthetic insecticides for pest control is the use of parasitoids of the genus Trichogramma. Considered the large number of agrochemical applications in grain yield, it is essential assess the selectivity of insecticides used, providing informations to assist in the maintenance of T. pretiosum in the area and can enhance the biological control of H. armigera, this being important for the establishment of integrated pest management in field crops. Therefore, the aim of this work was study the acute toxicity and the duration of the harm effects of insecticides on the parasitoid, study the effect of insecticides on the pupal stage of the parasitoid and determine the interference of insecticides on the parasitism capacity. Insecticides tested in laboratory conditions, Nomolt®, Avatar®, Belt®, Premio® and Intrepid® were considered innocuous to the adults. Lorsban® and Engeo Pleno® were harmful to the pupal stage, while Acefato®, Pirate® and Belt® although were considered innocuous they affect the next generations. Pirate®, Lorsban® and Engeo Pleno® interfered on the parasitism capacity, and Acefato® have a deleterious effect on the generation that has contact with the insecticide residue. The insecticides Nomolt®, Avatar®, Belt®, Acefato®, Premio® and Intrepid® were classified as short live (class 1). Lorsban® was moderately persistent (class 3) and Engeo Pleno® and Pirate® were classified as persistent (Class 4). Therefore, for an effective IPM program, it is important to use the selective insecticides. Insecticides that were harmful to the parasitoid adults could be used in the field, provided that the parasitoids do not occur naturally and the chemical applications do not coincide with the parasitoids release.

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No description available.

The nucleopolyhedrovirus of Epiphyas postvittana (EppoMNPV) is being studied as a potential biological control agent for leafroller insects in New Zealand. The aims of this project were for the identification of putative genes that are unique to, variant in or missing from, the EppoMNPV genome and the subsequent analysis of at least one of these genes. The purpose of this was to identify and characterise genes potentially involved in the specific host range and virulence of EppoMNPV. This was achieved in two steps. Three genome regions lacking linearity between EppoMNPV and the closely related OpMNPV were previously identified and the targeted sequencing of these three regions was the first aim of this project. The collation of the entire genome sequence of EppoMNPV and comparison to the genome sequences of 22 other baculoviruses completed the identification of genes that are unique to, variant in or missing from EppoMNPV. The EppoMNPV genome was found to be 118,584 bp in size encoding 136 putative proteins. A total of 29 genes were found to be common to all baculoviruses, while the lepidopteran baculoviruses share a total of 62 genes. The genome of EppoMNPV encodes four putative unique genes, the sequence of which offers no clues as to possible function. EppoMNPV lacks a homologue of the superoxide dismutase gene common to all other lepidopteran baculoviruses The EppoMNPV IE2 homologue was identified as a 311 amino acid protein with a truncation in the N-terminal region. We hypothesised that this truncation would lead to a loss of function, which could contribute to the virulence and/or host range of EppoMNPV For this reason, the characterisation of the EppoMNPV IE2 was taken up as the second part of this project. A comparative study between the AcMNPV and EppoMNPV IE2 proteins identified no differences in function between these two proteins in Sf21 cells. The EppoMNPV IE2 was capable of trans-activating three constitutive promoters and localised to discrete nuclear bodies. Cell cycle arrest was not achieved by either IE2 protein in our cell culture system. The role of four sequence motifs common to all IE2 proteins was studied with the aid of mutational analysis. Mutation of arginine and acidic rich sequences of EppoMNPV IE2 showed only a slight decrease in trans-activation activity while mutation of the RING-finger and coiled-coil motifs reduced trans-activation to less than half that of wild type IE2. Mutation of the coiled-coil motif resulted in reduced amounts of protein localising to discrete nuclear regions. A series of deletion mutants from the N- and C-termini of EppoMNPV IE2 identified that the C-terminal 111 amino acids of EppoMNPV IE2 was sufficient for nuclear targeting. Deletion of the C-terminal 19 amino acids resulted in an IE2 mutant completely defective in both localisation and transactivation. This demonstrates that localisation to discrete nuclear regions is essential for EppoMNPV IE2 to act as a transactivator.

The common brushtail possum (Trichosurus vulpecula) is the most significant vertebrate pest in New Zealand as an ecological threat to the indigenous biodiversity and an economic threat as a vector for bovine tuberculosis. Biological control is considered to be the most accepted management strategy to reduce the population, specifically by impairing fertility. Successful development of a biocontrol agent (most likely a protein or peptide macromolecule) requires identification of a compound that is species-specific and potent. The challenge is also to deliver the bioactive to this free-ranging, widespread, feral animal and ensure sufficient bioavailability. Macromolecules have low oral bioavailability, thus new formulation strategies are required to enhance stability and absorption in the gastrointestinal tract (GIT) of T. vulpecula. Oral administration of the bioactive contained within a non-toxic bait is the most practical delivery strategy. Essential to designing an oral delivery system is to quantify the transit time of different sized delivery systems. The gastrointestinal transit in T. vulpecula was investigated (n = 72) by gamma scintigraphy. Technetium-labelled (99mTc) anion exchange resin particles (75 - 125 (mu)m or 500 - 700 (mu)m) or solution (99mTc-DTPA) was administered orally. After 3, 6, 12, 24 or 32 h, distribution of radioactivity in excised GITs was determined. Transit profiles were similar for each formulation. For delivery to the hindgut, bioactives need protection for 12 h though the upper GIT. Particulate formulations may be retained in the caecum for up to 32 h. Transit time was not different between animals dosed in the evening or the morning. Furthermore, GIT morphology is different between specimens in this study from southern New Zealand and Australian specimens. This may reflect improved diet quality in New Zealand. A model protein (insulin) was incorporated into poly(ethyl 2-cyanoacrylate) (PECA) nanoparticles prepared by interfacial polymerisation of water-in-oil microemulsions. The mean size of nanoparticles was 220 nm with a mean entrapment efficiency of 78%, determined using reverse phase HPLC. In vitro release of insulin from PECA nanoparticles in phosphate buffer (0.067 M, pH 7.4) at 37°C was triphasic and not all entrapped insulin was released. Following in vitro incubation of nanoparticles with enzyme solutions prepared from the GIT of T. vulpecula, lumen enzymes were more aggressive towards insulin compared to mucosal enzymes and the hindgut lumen was the GIT region with the lowest degradation. For the first time in a marsupial species, the in vivo pharmacokinetics of insulin-loaded, PECA nanoparticles were investigated following i.v. and intra-caecal administration and measured by radioimmunoassay. The low cross reactivity of human and endogenous brushtail possum insulin means that T. vulpecula is a suitable non-diabetic model to study pharmacokinetics of insulin. The i.v. pharmacokinetics of insulin solution and insulin-loaded nanoparticles were similar. On intracaecal dosing, co-administration of a permeation enhancer (EDTA) resulted in a small increase in plasma insulin concentration compared to insulin-loaded nanoparticles alone. In conclusion, transit time to the caecum of T. vulpecula following oral delivery was 12 h for fluid and particulate formulations < 1 mm diameter and was independent of the time of day the dose was given. T. vulpecula is a potential non-diabetic model for the study of insulin pharmacokinetics. This thesis demonstrates the potential application of oral peptide and protein delivery technology in the area of wildlife management.