DISSERTAÇÃO - MBT Programa de Pós-Graduação em Biotecnologia e Recursos Naturais da Amazônia
URI permanente para esta coleçãohttps://ri.uea.edu.br/handle/riuea/2048
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Resultados da Pesquisa
Item Suscetibilidade a inseticidas, análise do gene Kdr (knockdown resistance) e de enzimas de detoxificação no mecanismo de resistência, em populações de Aedes aegypti da cidade de Manaus(Universidade do Estado do Amazonas, 2012-10-26) Silva, Eliane Christine Santos; Santos, Joselita Maria Mendes dos; Santos, Joselita Maria Mendes dos; Souza, Antônia Queiroz Lima de; Rodrigues, Iléa BrandãoContinued use of chemical pesticides and has led population to intense selection pressure, causing problems for vector control programs. Resistances Kdr (Knockdown resistance) have been reported in several locations in Brazil and other tropical and subtropical areas of the world, characterized by changes in sodium channel of insects, causing loss of coordination, after exposure to the insecticide, with recovery its effect often caused by recessive mutations, among which, Val1016Ile, located on voltage-gated sodium channel mosquito species A. aegypti (AaNav), dengue vector. This study aimed to evaluate the susceptibility of third instar larvae to the insecticide temefós, formulation ABATE 1G and adults, deltamethrin, and determining esterase activity, phosphoglucomutase and hexokinase, and the presence of the mutation Val1016Ile in four A. aegypti populations Manaus / AM, submitted and not submitted to insecticides. Were analyzed by means of bioassays of insecticide susceptibility, neighborhoods A. aegypti populations: Alvorada I, Coroado I, Monte das Oliveiras and Praça 14 de Janeiro, in decreasing order to estimate mortality rates, using the diagnostic doses of ABATE 1G (0.012 mg / L) and deltamethrin (5μg). With the results of bioassays were identified Praça 14 and Alvorada I populations like being the more resistant, and Coroado I and Monte das Oliveiras, less resistant. Unable to associate the esterase activity with resistance to organophosphate temephos, nor to deltamethrin, although they have been observed more bands (isoenzymes) in field populations when compared to the susceptible strain Rockefeller, submitted to the pyrethroid insecticide. In this case, there is possibility of esterases are being regulated by a genetic mechanism that insecticide occurs in the presence of preferential activation of certain isoenzyme form.The presence of Kdr mutation was found in all populations studied, and the Praça 14 population was the one with the highest frequency (0.98) and the Monte das Oliveiras, the lowest (0.58). The X2 values for the four populations were not significant, indicating that they are in Hardy-Weinberg equilibrium, with an excess of heterozygotes interpopulation and moderate genetic differentiation, and Praça 14 and the Monte das Oliveiras populations, the most distant genetically. The presence of the mutation Val1016Ile in all populations analyzed suggests that the use of pyrethroids is exposing populations to intense selection pressures. Thus, it is necessary to review the measures used for vector control, as well as the substitution of alternative insecticides deltamethrin by, making sure that the susceptibility is constantly monitored to ensure population control of this vector. Keywords: Aedes aegypti – resistance – organophosphates – pyrethroids – Kdr (Resistance)Item Prospecção, caracterização e purificação de lipase microbiana(Universidade do Estado do Amazonas, 2012-06-23) Gama, Auricelia Matos da; Yamane, Tetsuo; Willerding, André Luiz; Yamane, TetsuoLipases are enzymes which catalyze the synthesis and hydrolysis of glycerides in triacylglycerols and fatty acids. Its use as a biological catalyst has taken various applications in the pharmaceutical and food industries. Your source may be either animal, plant and microbial. With great emphasis on the latter due to the advantages as a means of obtaining, handling and great diversity, which makes filamentous fungi an interesting source of lipases. The search for new sources provides several jobs with specific objectives. Thus the aim of this study was to evaluate and select among two collections of filamentous fungi, potential producers of lipases. For this study, 316 strains were actually tested for lipolytic activity in liquid culture medium inducing lipase (olive oil). Of this total, 298 (94%) were positive and their lipase activity observed from the hydrolysis reaction with the substrate p-NPP with reading in the spectrophotometer at 410 nm. Only isolates exhibiting activity above 7000 U / ml were selected. Thus, only six isolates showed activities considered good and underwent further testing with enzyme activity in palm oil. The most promising strain was identified through molecular techniques as Endomelanconiopsis endophytica, producing an activity of 7895 U/mL for the olive oil and 10,727 U/mL for palm oil. The crude extract was then purified by ion exchange chromatography, resulting in three enzymes active form (two of 60 and one of 46 kDa) quantified by SDS-PAGE. And their identity verified by mass spectral and N-terminal sequencing by Edmann degradation. Two of them were presented blocked and the 46 kDa its sequence was obtained. Thus the study sought to contribute to the discovery of new lipases with biotechnological potential, contributing to a future database protein. Keywords: lipases, filamentous fungi, submerged fermentation, purificationItem Propagação Vegetativa do Protium spp: Protium heptaphyllum, Protium spruceanum e Protium guacayanum(Universidade do Estado do Amazonas, 2012-04-20) Lima, Suelen Cristina de Sousa; Sampaio, Paulo de Tarso Barbosa; Quisen, Regina Caetano; Sampaio, Paulo de Tarso BarbosaThe production of secondary metabolites by plant cell culture contributed to the progress in various areas of plant physiology and biochemistry. However, a restricting factor for the definition of protocols to in vitro cell culture oil-producing species such as Protium Heptaphyllum, is contamination of the nutrient medium by microorganisms. This contamination is established in the media and/or in vegetable material, which can be toxic for culture by competing for nutrients, producing toxic substances and inhibiting development of the explant, causing its loss. Thus, , this study aimed to test methods of disinfection for establishment in vitro leaf explants of P. heptaphyllum. Assays were performed at the Laboratory of Tissue Culture Plants at Embrapa Western Amazon, Manaus, Amazonas. For surface disinfection, leaves of P. heptaphyllum, agents were tested in the control of microorganisms: Plant Preservative Mixture (PPM®), Cercobin® with Agrimicina® and essentials oils. It was observed that the PPM® at 1% and 2% was more effective in controlling infection of leaf explants. For the assays with essential oils was obtained control of microbial contaminants, however the concentration used was toxic to the tissue. In view of these results, we suggest new tests with other disinfesting agents, antioxidants for this species. Key words: plant cell culture, in vitro establishment, tropical forest species, micropropagation.Item Potencial fungicida de extrato foliar de Cariocar Villosum (AUBL.) Pers (Caryocaraceae)(Universidade do Estado do Amazonas, 2012-03-29) Valente, Paula Mara Rodrigues; Scudeller, Veridiana Vizoni; Silva, Ademir Castro e; Scudeller, Veridiana VizoniThe use of fungicides is in many cases the only viable and effective measure to control diseases and ensure the sustainability of agriculture. On the other hand, it is also a technology that brings negative impacts to the environment and public health. The fungus Fusarium sp. cause severe damage to crops of vegetables in the city of Parintins - Am is considered soil pathogen capable of causing severe damage to many economically important agricultural species. In this context, the species Caryocar villosum (piquiá) appears as a promising possibility because preliminary studies reveal the existence of substances with fungicidal and / or fungistatic. This study aimed to obtain crude extracts ethanol, methanol and aqueous leaves C. villosum, and analyze their activity on the fungus Fusarium sp. Using two modes of extraction, hot and cold. From leaves of lettuce necrotic been made to isolate Fusarium sp. Which was cultivated in BDA. Foram The effects of the extracts on o crescimento mycelial concentrations 1g / L, 5g / L, 10g / L and 15g / L measuring the diameter of fungal colonies for seven days. For counting was used ten cellophane disks of 0.8 cm in diameter which were placed in petri dishes on three discs of filter paper soaked in solution at concentrations of extracts 1g / L, 5g / L, 10g / L and 15g / L in triplicate. At concentrations of 10 and 15g / L of ethanol extract hot, significant inhibitions were observed in mycelial growth of the fungus. Concentrations 5, 10 and 15 g / L of hot aqueous ethanol extract and significant inhibitions were observed on conidial germination of Fusarium held from 16h incubation. Key words: Caryocar villosum, a fungicide, Fusarium sp.Item Microbiota Bacteriana associada às esponjas de água doce da Amazônia(Universidade do Estado do Amazonas, 2012-07-15) Costa, Glaucia Cristina Manço da; Rodrigues, José Carlos Verle; Procópio, Aldo Rodrigues de Lima; Rodrigues, José Carlos VerleSponges are multicellular organisms classified in the phylum Porifera, been considered the most primitive metazoan. They present a cellular structure without true tissues. They live in tropical oceans, temperate and polar regions, and some species inhabit fresh water bodies. Sponges provide particular microenvironment conditions within their tissues. It allows them to the harboring of a diverse microbial community, which will be generating secondary metabolites. Those metabolites represent a source to search for new compounds of biotechnological interest. The ecological study of these microorganisms permits a better understanding of interaction dynamics and processes occurring in the environment. Different strategies could be applied to the analyze of genetic diversity of microorganisms in an environmental sample. Bacterias are key participants on natural ecosystems, mainly due their metabolic variability and adaptable physiology. Biocatalysis or biotransformation is the technique that uses biological catalysts to convert a substrate. It promotes chemical changes using enzymes (isolated or as parts of live microorganisms), The technology is competitive and doesn‟t harm the environment and is able to produce bioactive compounds with high enantiomeric purity. This work aims to characterize the microorganisms (bacteria) associated to freshwater sponges from two different Amazonian locations. The sponges were collected in the vicinities of the cities of Parintins and Manaus. Bacterias were grown and purified in selective medias. DNA extractions were made and sequencing of a 16S rDNA gene fragment performed for identification fingerprinting. The bioreduction reaction was performed with three bacterial strains and three substrates, p-nitroacetophenone, acetophenone and 3-hydroxy-2-methylene-3-(4'-nitrophenyl) methyl propanoic acid. The reaction progress was monitored by Gas Chromatography (GC) and Gas Chromatography Mass-Spectrometer (CG-MS). Hundred isolates were obtained from samples collected from Parintins and 117 isolates from Manaus. From these isolates, 31.5% were identified as belonging to Bacillus genus. Regarding the bioreduction reactions, the three bacterias showed the same retention time (13 minutes), and the reduction was considered as chemoselective, because bacteria reduced the nitro group present in amine. To our knowledge this is the first report of communities of bacteria associated to Amazonian fresh water sponges performing selective bioreduction. This work provide initial basis to further investigations of microbial communities in freshwater sponges in the Amazon region. Keywords: sponge – freshwater – bacteria - biocatalysisItem Investigação de atividades antimicrobiana de extratos de plantas utilizadas na medicina popular por moradores de comunidades rurais de Manacapuru - AM(Universidade do Estado do Amazonas, 2012-05-12) Silva, Daniele Lobo da; Brocki, Elisabete; Souza, Antonia Queiroz Lima de; Brocki, ElisabeteThere is a great number of people that are concerned about health and quality of life everyday, seeking in plants an alternative way of curing diseases, through the use of medicinal plants with these properties. Trichophyton rubrum is a dermatophyte fungus, that has the ability to invade keratinized tissue of humans and other animals, causing infections called dermatophytosis. The bacterias Pseudomonas aeruginosa and Staphylococcus aureus are opportunistic agents that are associated with several human infections, especially in nosocomial settings. The aim of this study was to investigate the antimicrobial activity of plant extracts used in folk medicine by residents of rural communities of Manacapuru-AM. Ethnobotanical data were collected through interviews that resulted in the selection of five plant species, using Importance Value (IVs) method to obtain ethyl acetate and ethanol extracts by maceration, and the aqueous by infusion. Antimicrobial susceptibility tests were performed using agar diffusion method, and ethanol extracts of Momordica charantia and Vatairea guianensis presented antimicrobial activity against P. aeruginosa, with minimum inhibitory concentration of 250 g / ml and 125 mg / ml, respectively. T. rubrum was not susceptible to the studied extracts. Keywords: Ethnobotany, Medicinal plants, Trichophyton rubrum, Staphylococcus aureus and Pseudomonas aeruginosas.Item Fungos amazônicos com potencial para degradar Chorume in natura obtido do lixão municipal de Parintins(Universidade do Estado do Amazonas, 2012-04-26) Nunes, Adriana da Silva; Silva, Ademir Castro e; Silva, Ademir Castro e; Rodriguez, Arelis Abalos; Perez, Enrique Ramon MolinaThere are several techniques for treating toxic effluents as manure, but the white rot fungi have achieved notoriety in recent decades in fits of continuing environmental degradation. But little is known about the fungi of the Amazon region and their behavior in culture medium supplemented with toxic compounds. The aim of this study was to evaluate the potential of fungi in the degradation of Amazonian fresh manure obtained from the municipal landfill Parintins / AM. The strain Pycnoporus sanguineus and FV-12 and were inoculated into culture media different to the first glucose and different concentrations of slurry (M1) and second with the addition of inducer seringaldazine (M2). The strains grew up in different culture media, P. sanguineus obtained the highest mycelial growth in M2 medium, while FV-12 stood out in the middle of M1. However, other nutritional and physiological factors affecting the growth of fungi in a medium containing toxic compounds. P.sanguineus produced a larger amount of biomass is increased as the concentration of the slurry (1%, 2%, 4%) in the middle, however, the opposite behavior is observed in treatment with the fungus FV-12. With regard to the degradation of total protein FV-12 had higher percentages, but smaller in relation to inhibition of toxicity and discoloration, that effect P.sangineus said fungus is possibly by the formation of byproducts resulting from the more toxic action of the fungus FV-12. This in turn, increased levels of excreted MnP and LiP within thirty days, noting combined action of these enzymes, which may have influenced the further degradation of total protein per FV-12. In relation to the enzyme laccase were detected low levels of activity in the periods tested for P. sanguineus and FV-12, possibly due to influences of factors such as incubation time and carbon source. The fungus P.sanguineus had the highest percentage of degradation of manure, and their possible application in the treatment of manure. Keywords: Slurry, Fungi Amazon, Degradation, Oxidative Enzymes, Parintins/AM.Item Fungos Amazônicos com potencial para degradação de Polietileno da Amazônia(Universidade do Estado do Amazonas, 2012-04-26) Soares, Elaine Pires; Silva, Ademir Castro e; Silva, Ademir Castro e; Cordeiro, Milade dos Santos Carneiro; Rodriguez, Arelis AbalosThis work aims to study the degradative action of the synthetic polymer polyethylene terephthalate - PET by basidiomycetous fungi Amazon. Three different fungal strains (P. sanguineus, FBL, FV12) and the consortium between them in stationary liquid culture medium, with or without supplementation with flour bark tucumã at 15, 30, 60 and 90 days of growth at 27 ° C +1. Samples of PET were subjected to pre-treatment temperature 35 ° C and 50 ° C. After incubation were evaluated polymer mass loss, the determination of ligninolytic enzymes and analysis of samples using PET scanning electron microscopy to verify the occurrence of morphological and / or structural polymeric surface. All the strains resulted in loss of mass of PET, the highlights were the fungi FBL and FV12 and the consortium C1-PYC + FBL + FV12 within 90 days of cultivation, with PET subjected to the pretreatment temperature 50 ° C, with greater weight loss plastic. All fungi tested showed enzyme activity lignolítica. The manganese peroxidase (MnP) was the enzyme that showed the highest activity among the three enzymes studied. The strain FV12 PET subjected to temperature 50 ° C in 90 days of incubation showed the highest mean value of this enzyme. It should be noted that the best results were obtained with a culture medium supplemented with tucumã shell flour. shows that PET subjected to temperature 50 ° C, suffered severe wear of the polymeric surface with flaking, ruptures and cracked appearance across the surface, as well as bleaching of the sample. It follows that the Amazon fungi studied have potential to degrade the Polyethylene Terephthalate. Keywords: Fungi Amazon, Polyethylene Terephthalate, oxidative enzymes, biodegradation.Item Estudos de QSAR-2D em quinolina metanol com atividade antimalárica frente ao Plasmodium falciparum(Universidade do Estado do Amazonas, 2012-06-11) Duarte, Carla dos Santos; Marques, Alberto dos Santos; Takahata, Yuji; Marques, Alberto dos Santos; Nunez, Cecilia Veronica; Rodrigues, Déa BrandãoAn analysis of 2D-QSAR was realized in twelve two quinoline methanol substituted in position 4 with antimalarial activity against the Plasmodium falciparum. The semi-empirical quantum methods AM1 and RM1 were used to calculate the geometry and some molecular properties. The DRAGON program was also used to produce descriptors and MobyDigs was used to select descriptors and build QSAR models. The best obtained from QSAR models used multiple linear regression. The model with 12 composed have descriptors, and G3v R2e, resulting in r2 = 0.90, q2 = 0.79 and q2boot = 0.72. The validation was performed on RQK fitness functions. It was shown that the QSAR model satisfies all the criteria required for validation, indicating that the regression model is acceptable. The external validation was performed with the exclusion of compounds AAQM_4, AAQM_7 and AAQM_11. The new model was generated as descriptors, and Mor28u MPC05, with r2 = 0.98, q2 = 0.97 and q2boot = 0.94. Considered valid for the functions RQK adjustment. Keywords: malaria, QSAR, quinoline methanol.Item Estudo Espectroscópio experimental e Teórico do novo mutagênico /carcinogênico aminofenilnorharmana(Universidade do Estado do Amazonas, 2012-06-13) Reis, André Henrique Araujo; Marques, Alberto dos Santos; Marques, Alberto dos Santos; Nunez, Cecilia Veronica; Pereira, Luiza GabrielThe UV / visible norharmana (9H-pyrido [3,4-b] indole) (NH) and aniline dissolved in ethanol were studied. The NH is found in cigarette smoke in high concentrations, cooked meat, cooked fish, and was detected in the urine of people and in breast milk, and is also found in some plants of the Amazon, for example, chacrono (Psychotria viridis) , and is known for its active principle hallucinogen. The aniline is present in cigarette smoke in concentrations too high, and is an important raw material in the chemical industry. Were calculated excitation energies, forces the oscillator to NH, and aniline (9-(4'-Aminophenyl)-9H-pyrido [3,4-b] indole) (APNH) with the Density Functional Theory dependent time using the B3LYP functional and basis set 6-31G *. The spectra of the molecules were simulated and compared with the experimental spectra to analyze and interpret the observed spectra. The mutagenic / carcinogenic of APNH was discussed based on the RHF/6-31G * ab initio calculations made to determine the molecular structure and atomic charges Key words: APNH, UV absorption spectra, Time Dependent Density Functional Theory -TFDDT, ab inítio method.