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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Propagação vegetativa de crateva benthamii com diferentes concentrações de ana
(Universidade do Estado do Amazonas, 2004-11-24) Silva , Ângela Alves da; Maia , Lúcia Alencar; Yuyama , Kaoru; Maia , Lúcia Alencar; Canto , Acilino do Carmo; Silva, Ademir Castro e
Crateva benthamii Eichler (Capparaceae) is a typical floodplain forest species with high economic potential on pisciculture, pharmacological industry and environment recuperation in degraded areas. This work assessed the C. benthamii propagation in different ANA concentration and the kind of stake. The experiment was installed in the INPA warren V8 (Manaus-AM, Brazil). It was utilized delineation of casual blocks with five repetitions, with 3 x 4 factorial scheme, in which the factors comprehended the kinds of stake (apex, middle and base) and ANA concentration (0, 100, 1.000 and 10.000ppm). The bases of the stakes were plunged into ANA aqueous solutions during 24 hours. The evaluated parameters were: percentage of survival, percentage on rooting and cuttings having shoots and roots. After 90 days of planting, the apex cuttings showed lesser percentage on survival with 100% of mortality in 10.000ppm treatment. Between middle stakes and base stakes there was no statistical difference, showing higher indices on survival (70% and 80%, respectively). The higher percentage of rooting was in 1.000ppm concentration (70%), there was no statistical difference when compared with stakes any hormone treatment and stakes with 100ppm. The middle stakes and base stakes showed the highest percentages of shoot protrusion at 90 days (38,75% and 36,25%). There was statistical difference when compared with apex stakes (17,50%). The middle stakes and the base stakes of C. benthamii showed the highest percentage of survival. The variation concentration about ANA tested did not favor the rooting. C. benthamii shows higher potential for propagation per cutting and can be an option to recuperate degraded areas in floodplain forest.
Óleo volátil e antioxidantes de folhas de virola michelii heckel (myristicaceae)
(Universidade do Estado do Amazonas, 2004-12-15) Torres, Zelina Estevam dos Santos; Yoshida, Massayoshi; Yoshida , Massayoshi; Pinheiro , Maria Lúcia Belém; Nunez , Cecilia Veronica
This work describes the phytochemical study of Virola michelii Heckel that occur in the Amazon Region popularly known as ucuúba-preta and its leaves are used in the popular medicine as hot poultice on the skin, to treat infections caused by fungus. The fresh smashed leaves of Virola michelii submitted to the hydrodistilation afforded a volatile oil, which analyzed through coupled system gas chromatograph to mass spectrometer evidenced the occurrence of one monoterpene and of seventeen sesquiterpenes. The ethyl alcohol extract of V. michelii leaves was fractionated by ethyl acetate and aqueous methanol partition, followed by solvent evaporation of the fractions, yielded the respective residues. The ethyl acetate residue submitted to chromatographic techniques, allowed the isolation of three compounds, two furofuran lignan, “eudesmin” (rel- (1S,2R,5R,6S)-2,6-di-(3,4-dimethoxyphenyl]-3,7-dioxabicyclo[3.3.0]octane) and “phillygenol” rel-(1S,2S,5R,6R)-6-(4-hydroxy-3-methoxyphenyl)-2-(3,4-dimethoxyphenyl)-3,7- dioxabicyclo[3.3.0]octane and one phytosterol, the β-sitosterol. The structural determination was based on the interpretation of the 1 H and 13C nuclear magnetic resonance spectra and mass spectra. The isolated furofuran lignans were submitted to test for antioxidant activity with DPPH radical followed by photometric detection. It was observed that the phillygenol was more active than eudesmin, in the assay which quercetin was used as standard.
Desenvolvimento de estacas de crajiru (Arrabidaea chica Verlot.) em função do diâmetro da estaca e do ambiente de cultivo.
(Universidade do Estado do Amazonas, 2005-03-31) Felix , Francisco Cleber; Sampaio, Paulo de Tarso Barbosa; Pohlit, Adrian Martin; Sampaio , Paulo de Tarso Barbosa; Costa, Suely de Souza; Chaves, Francisco Celio Maia
The goal of this work was to observe the effects of two growth environments on two types of crajiru (Arrabidaea chica Verlot., family Bignoniaceae) as a function of cutting diameter ultimately to establish the better type of crajiru (type 1 or 2) for propagation from cuttings. The experiment was conducted using an entirely randomized method with a 2 x 2 x 4 factorial scheme, with 6 repetitions, 2 root development environments (1 – substrate and 2 – water) and 4 average basal cutting diameters: 1.1, 0.6, 0.4 and 0.2 cm, designated woody, semi-woody, herbaceous and apical stems, respectively. The cuttings were approximately 15 cm in length having leaves only on apical cuttings and 5 cuttings per repetition. The substrate was prepared from area : argila : humus (3 : 1 : 1). Each cutting was placed in an individual poliethylene bag of 1 kg capacity. For cuttings in liquid environment, 2 L polyethylene terephthalate bottles (PET) containing approximately 500 mL of water were used. From the start of the experiment, the cuttings were maintained in an environment covered with transparent plastic roofing and received daily irrigation. After 90 days, the following variables were evaluated: % established cuttings, average length of new growth (cm), average length of roots (cm) dry mass of leaves and roots (g / plant). After variance analysis, the averages were compared using the Tukey test and a 1 % probability. Crajirú responded well to vegetative regeneration from cuttings and the type 2 variety developed better in general than type 1. The percentage of established plants was greater in the substrate 1 (soil) for cuttings of average diameter 1.1 cm. Other variables were also superior for this diameter cutting, and the substrate was responsible for the nutirition of these cuttings, which presented better development in those of 1.1 cm diameter. Since the difference between the development in both environments (substrate vc. Water) was small, and the interaction treatment x environment x diameter was not significant, one can site an economical advantage using water for the preparation of crajirú plants from cuttings.
Propagação in vitro de Andiroba (Carapa guianensis Aublet)
(Universidade do Estado do Amazonas, 2005-04-28) Brandao, Helio Leonardo Moura; Sampaio, Paulo de Tarso Barbosa; Sampaio, Paulo de Tarso Barbosa
The goal of this work was to develop a protocol for cloning through somatic embryogenesis of Andiroba (Carapa guianensis Aublet). A species abundantly found in north Brazil utilized for medicinal and cosmethic purposes and as a source of compounds that may be used to develop new synthetic drugs. To promote somatic embryogenesis were tested NAA, BAP, 2.4-D, TDZ and IBA, in different concentrations and different combinations in explants obteined of young plants trated with Benolmyl. The explants were cultivated in a medium containing MS salt and vitamins (Murashige & Skoog, 1962) being the mineral salts reduced to half strength and added with myo-inositol (100 mg.L-1), sucrose (30 g.L-1) and agar (8.0 g.L-1), and the pH was adjusted to 5.8. The cultures were inoculated and then kept at a 16 hour-photoperiod, temperature of 25 ± 2ºC and 25µmoles.m-2.s-1 radiation for 30 days. Three different sources of explant were tested: apical, leaf buds and leaves. The source that showed best resoults of rooting was the leaf buds trested with NAA 10,0 mg\l. As well, leaves rooting showed best resoults treted with NAA. However apical source showed high level contamination, fact that took the increasing of a new protocol of assepsy. The results showed that it is possible to obtain high quantities of shoots and then plantlets which could be cultivated in vitro through the methods tested in this experiment. Apical source appear as the best way to obtain a plant of Andiroba by somathic embriogenesis, NAA promotes the rooting in leaf buds a leaves and BAP promotes shoots in apical explants. Key words: 1.Micropropagation, 2.Andiroba, 3.Hormonal combination and 4. growth regulators
Análise de marcadores químicos do produto fitoterápico Dermodilapiol (Piper aduncum)
(Universidade do Estado do Amazonas, 2005-05-19) Oliveira, Laura Cristina Pereira de; Nunomura , Sergio Massayoshi; Nunomura , Sergio Massayoshi; Pinheiro, Maria Lúcia Belém; Lima , Maria da Paz
This work describes the isolation and structure elucidation of two phenylpropanoids (dillapiol and myristicin), one prenylated benzoic acid derivative (methyl 4-hydroxy-3-(3-methyl-2-butenyl)-benzoate), one chalcone (dihydroflavokawin B) and one flavanone (sakuranetin) from the phytoterapeutical product Dermodilapiol, produced from the aerial parts of Piper aduncum. The isolation of the chemical markers has been done from its active fraction, obtained from a liquid-liquid partition and further chromatographic steps. Four different lots of the product have been analysed by high performance liquid chromatography using two different methods. A longer method was used to describe the chromatographic profiles of each lot and a shorter method was used to quantify two of the chemical markers in the product. In both cases, it could be observed a correlation between the chemical composition and biological activity. It has also been studied the essential oil production of leaves of P. aduncum collected in three different places in Manaus for a period of one year. During this period, dillapiol has always been the major component. A small chemical variability was observed to all specimens studied and apparently was correlated to the rainy season of the city.
Atividade inseticida de palicourea marcgravii st. Hil. (rubiaceae) e piper aduncum l. (piperaceae) sobre cigarrinha (aetalion sp.), praga de importância econômica no Amazonas
(Universidade do Estado do Amazonas, 2005-09-26) Silva, Wilson Castro; Ribeiro , Joana D’Arc; Ribeiro, Joana D’Arc
The use of extracts of toxic and medicinal plants, usually under exploited in Amazonia, is an alternative to control phytophagous insects due to their low costs, easy preparation, use and environmental safety. The lack of papers about the insecticidal activity of Palicourea marcgravii and Piper aduncum on treehopper (Aetalion sp.) was the reason to carry out this research. The aim of this work was to evaluate the insecticidal action of P. marcgravii and P. aduncum on Aetalion sp. Adult insects were collected on the tree Clitoria fairchildiana, separated in groups of ten individuals into plastic vessels and exposed to topic application of aqueous extracts of leaves and roots of both plant species. The extracts of leaves and roots of P. marcgravii were applied at the concentrations of 30, 40 and 50 mg/mL and 10, 20 and 30 mg/mL, respectively, whereas for P. aduncum the extracts were applied at the concentrations of 10, 20 and 30 mg/mL, for both extracts of leaves and roots. The control groups were treated with distilled water. After the treatments, the vessels were transferred to a greenhouse and maintained under natural conditions of temperature, light and humidity. The tests lasted 48 h and every 12 h the mortality rate was evaluated. The experiments were performed in a complete randomized design with three treatments in five replicates more a attest group. The data were evaluated using One-way Analysis of Variance (ANOVA) followed by a multiple comparison method (Dunnett’s test). The Probit method was used for calculation of the median lethal concentration (LC50) and the median lethal time (LT50) values, and their 95% confidence limits. The computer program TOXRAT® was used for the Probit analysis and to fit the dose-response curves. The extract of roots of P. marcgravii was the most toxic (LC50 = 12,4 mg/mL), but not statistically different from the P. aduncum extracts (leaves and roots, LC50 = 20,9 and 20,9 mg/mL, respectively). The extracts of P. aduncum were toxic in shorter exposure time (leaves and roots, LT50 = 30,2 and 22,3 h, respectively) than the P. marcgravii extracts (leaves and roots, LT50 = 41,2 and 34,9 h, respectively), but not statistically different. The extracts of both species showed insecticidal action on Aetalion sp., however, more detailed studies about the chemical composition of the extracts and their toxicity for non-target (beneficial) organisms should be done before to recommend them as insecticides.
Estudo de Marcadores Químicos de Ambrosia artemisiifolia L (Asteraceae)
(Universidade do Estado do Amazonas, 2005-10-24) Valdivino , Hyelen Bezerra Gouvea; Nunomura , Sergio Massayoshi; Nunomura , Sergio Massayoshi; Borras , Maria Rosa Lozano; Yoshida, Massayoshi
Ambrosia artemisiifolia L. (Asteraceae) is a annual specie, normally considered a weed, that grows well in tropical areas, including the Amazon region. It is popularly known in Brazil as “catinga-de-bode”, “artemisia”, “arquemijo” and “artemija”. Traditionally, the tea of A. artemisiifolia. leaves is used for the treatment of stomach illness. This work describes the isolation of the sesquiterpene lactones, ambrosin and damsin, for the first time in this specie. Aqueous and chloroform extracts were evaluated in three different models of gastric lesions (ethanol, stress and indomethacin) induced acutely and showed significant protective activity. Chromatographic fingerprints of aqueous and chloroform extracts by high performance liquid chromatography and also essential oil analysis by high resolution gas chromatography were obtained from four different individuals collected in different locations in the Amazon region, Iranduba (AM), Juruti (PA), Manaus (AM) and Itacoatiara (AM), in order to develop chromatographic methods to be used in the quality control of the isolated chemical markers, accordingly to the resolution RDC N. 48.
Estudo fitoquímico e avaliação da potencialidade farmacológica de extratos de crateva benthamii (capparaceae)
(Universidade do Estado do Amazonas, 2005-10-25) Pereira Junior , Raimundo Carlos; Maia , Lucia Maria de Alencar; Nunez , Cecilia Veronica; Maia , Lucia Maria de Alencar
Crateva benthamii (Capparaceae), a common tree species in the Amazonian floodplain areas, may reach from 6 to 12 m in height. There are reports on it being used by the people inhabiting the riverside, on account of its bark being used as antiseptic and against snake poison, its roots and leaves being used as tonics for stomachic problems and its topical use as auxiliaries in the recovery of the rheumatism. Aiming to evaluate the pharmacological and biotechnological potentialities of the C. benthamii extracts, the present study reports on the findings obtained on the evaluation of the methanol/water extract (80:20) of the C. benthamii bark, on the antifungal tests against the xylophagous fungus Pycnoporus sanguineus and on the methanol/water (80:20) and dichloromethane extracts of the bark on the antibacterial tests against species Staphylococus aureus, Escherichia coli , Salmonella london ,Shigella sonni, Shigella paratyphi , Shigella tryphimurium, as well as the characterization of the substances that compose the essential oil extracted of its leaves. The methanol/water extract (80:20) on the antibacterial assays showed to be active to Staphylococus aureus bacterium species on 1.20 mg/mL concentration both by plate cavity diffusion (CP) as plate discus (DP) method, while the dichloromethane extract did not show activity for others bacteria studied. On the assay to evaluate the antifungal extract activity against P. sanguineus the methanol/water extract on the concentration of 0,1 g/mL showed moderate inhibitor activity on micelial growth. Statistical data showed that the concentration increase may inhibit micelial growth. The 13C NMR spectrum analyses of the dichloromethane extract showed Lupeol and Germanicol as majority triterpenes. Chromatographic fraction resulted on the Lupeol isolation and identification in addition to another not identified substance similar to yellowish-coloured oil, with a pleasant odour, whose 13C NMR spectrum suggests being a -amirin ester. In the essential oil CG/EM analysis eight substances were identified, with the trans-caryophyllene being one of the major compounds.
Atividade enzimatica hidrolítica dos fungos amazônicos pycnoporus sanguineus (l.f.) Murr e panus crinitus (l.ex. Fr.) Singer
(Universidade do Estado do Amazonas, 2005-11-24) Carmo , Cynara da Cruz; Silva , Ademir de Castro e; Roland, Ivete de Araújo; Carmo, Cynara da Cruz; Cordeiro , Milade Carneiro; Lúcia Alencar, Maia
Enzymes of hydrolytic nature such as cellulase, xylanase and glicosidase and/or others agents non-enzymatic produced by wood decay fungi, show a huge biotechnological potential. I order to investigate hydrolytic potential of two amazonian wood decay fungi, Pycnoporus sanguineus (L.:F.) Murr and Panus crinitus (L. Ex. Fr.) Singer, studies on influence of pH, temperature and medium at growth of fungi as well as determination of enzymatic activity of FPA, -glucanase and -xylosidase on stationary and agitation conditions at 180 rpm were done. Fungi studied shown preference by acid conditions (pH optimum 5,0) and temperature optimum between 30- 35° C. P. sanguineus had these optimum conditions at sabouraud medium supplemented with peptone while P. crinitus at malt extract medium. Temperature had more influence on fungal growth. Enzymatic activity were better at agitation conditions. At stationary condition with no glucose in the medium it wasn't detected FPA activity in both fungi and only to P. sanguinues the endo--glucanse activity was detected. At stationary growth the addition of glucose increase about 38% endo--glucanase activity for P. sanguineus but it not contributed to increase this activity for P. crinitus. For both fungi the -xylosidase activity at stationary conditions and presence of glucose it wasn't followed by endo-glucanase activity. P. crinitus presents a greater hemicelulolytic activity become evident by greater -xylosidase activity while P. sanguineus showed great celulolitic activity. There is need studies to find new medium to maximize hydrolytic enzyme activity to get viable purification to possible biotechnological uses.
Propagação Vegetativa da Preciosa (Aniba canelilla (H. B.K) Mez) pelo Método da Estaquia
(Universidade do Estado do Amazonas, 2005-12-01) Siqueira , Jhassem Antônio Silva de; Sampaio , Paulo de Tarso Barbosa; Costa, Suely de Souza; Sampaio http://lattes.cnpq.br/0517307169401972, Paulo de Tarso Barbosa
The precious wood (Aniba canelilla (H.B.K.) Mez) is a native species from Amazon Region. It is a popular species well know in the region for its aromatic tea that can be obtained from their leaves, bark and trunk. It is a species with a great economical potential, because it can give a wood of high quality, it has also medicinal purpose, and nowadays it is being used by the cosmetic industry. The essential oil has a high concentration of nitrofeniletano (a natural odour). This work had the aim to establish a methodology for the propagation by rooting of precious wood species cuttings. Furthermore it aims to evaluate the action of different concentrations of indolbutyric acid (IBA), at 0; 300; 600; 1200; 2000, on t he development of the cutting that were obtained from juvenile and adult material. The experiment was developed in greenhouse with an intermittent mist system during a period of 180 days (around six months). The experiment was statically analysed by completely randomized method, the data were also submitted to analysis of variance and the means were compared by Tukey test analysis at 5% probability. The results indicated that it is possible to promote the vegetative propagation of precious wood by the cutting method, independent of the hormones concentration used at the experiment. The AIB use on the concentration of 2000 ppm favored the rooting (79,04%), the survival (89,43%) and sprouted (64%) of the cuttings coming from juvenile material after 180 days of the implementation of the experiment. The comparison of the means of fresh material weights of aerial part and root, and the weight of the dry material of the aerial part of the plant and also of the cuttings. They did not showed a significative difference among them. . Cuttings from adult material in the treatments here evaluated showed rooting below 1%, and a final survival means of 0,76.
Propagação vegetativa in vitro de Aniba rosaeodora Ducke (Lauraceae)
(Universidade do Estado do Amazonas, 2006-02-28) Jardim, Lyana Silva; Sampaio, Paulo de Tarso Barbosa; Sampaio, Paulo de Tarso Barbosa
Rosewood (Aniba rosaeodora Ducke), is a specie of great economic value for producing the essential oil linalol, used for perfume industry, causing intense exploration of the natural populations, and its disappearance in the areas of easy access. Like this, the purpose of this study was to establish a protocol for rosewood in vitro propagation, using apical and nodal segments and leaves, aiming at new techniques for specie’s reproduction. Firstly, the explants were submitted to differents process of disinfection, using solution of benomyl (4g.L-1), solution of sodium hipoclorite in 15% and 20%, with tween 20, in differents time exposure. After the explants were inoculated in MS medium, with 30g.L-1 of sucrose and 8g.L-1 of agar added to it , and they were left in dark conditions for 48 hours in B.O.D under 26ºC, and led to culture room. After, the apical and nodal segments, apparent healthy, were submitted to buds, roots and callus induction. They were transferred to culture medium containing growth regulators: BAP (0,0 e 4,0 mg.L -1), AIA, ANA and 2,4-D (0,0; 3,0 e 6,0 mg.L-1), and their respective combinations. The design was complete randomized in arranged factorial 9 X 2, with 18 treatments, each one with 15 replications. For the folial segments, were used: BAP, TDZ, ANA, AIA and 2,4-D (5,0 mg.L-1) to induction of foliar organogenesis. The design was randomized blocks consisted of 6 treatments, each one with 8 blocks e 2 replications. After 90 days, the plantlets took root in vitro were transplanted in plastic glass covered for transparent plastic sack, and evaluated in three kids of substratum for the acclimatization process. The first and the second substratum contained dark soil, clay and sand in proportions 2:1:1 and 1:2:1 (v/v) covered for vermiculite respectively; and the third substratum contained for vermiculite just. At the end of the study, the results indicated that the best process of disinfection for the apical and nodal segments was the use of benomyl solution (4g.L-1) during 24 hours, following the solution of sodium hipoclorite in 20% with tween 20, during 20 minutes, with 81,7% of survival of the explants. For the buds induction, the treatment containing 4,0 mg.L-1 of BAP + 6,0 mg.L-1 of AIA, showed the best, with the average of 1,50 buds/explant, followed by the treatment with 4,0 mg.L-1 of BAP + 6,0 mg.L-1 of ANA, which the average was 1,45 buds/explant. For the rooting induction, the best medium was containing 3,0 mg.L-1 of ANA, showing 1,68 roots/explant, followed by the treatment with 6,0 mg.L-1 of ANA, showing an 17 average of 1,39. For the callus induction, all the treatments formed callus, therefore, the medium containing 4,0 mg.L-1 of BAP + 6,0 mg.L-1 of 2,4-D, showed the best result, with 1,45 callus/explant, followed by the treatment with 4,0 mg.L-1 of BAP + 3,0 mg.L-1 of 2,4-D, of which an average was,1,40 callus/explant. For the folial segments, the best disinfection was the use of benomyl solution (4g.L-1) during 20 hours, followed by the solution of sodium hipoclorite in 20% with three drops of tween 20, during 15 minutes of exposure, showing 80% of survival of the explants. For the folial organogenesis, both treatments, F3 containg 5,0 mg.L-1 of ANA and F5 containg 5,0 mg.L-1 of 2,4-D, presented the best results for the callus induction, with an average of 1,4 callus/explant (both treatments). For the acclimatization process, none of the substratum presented efficiency for the plantlets adaptation in the new environmemtal conditions. The survival percentage of the explants for all the treatements was zero. In less of 30 days after the transplant, the plantlets presented complete necrose with partial lost of the leaves, and appearance of fungi in some replications. Key words: 1. Aniba rosaeodora Ducke; 2. In vitro propagation; 3. Growth regulators.
Atividade Enzimática da Lacase em Três Fungos Amazônicos Degradadores de Madeira
(Universidade do Estado do Amazonas, 2006-03-12) Silva, Ezequias Andrade da; Silva, Ademir Castro e; Silva, Ademir Castro e
Fungi belong to basidiomycete class have been confered efficiency in the biodegradation process of ligninocelullosic materials due to their enzymes. These enzymes have commercial potential that could be utilized to many industrial applications. This paper evaluate the micelial growth and enzymatic activity of laccase of three amazonic white-rot fungi (Pycnoporus sanguineus, FSF7 (Trametes sp) e FSF11 (Trametes sp). Micelial growth was evaluated through the micelial progress in solid media BDA with different concentrations (30, 90, 120 and 300 mcg) of biotin. To enzymatic activity of laccase was used liquid medium (agar-malt) with different concentrations of glucose/ yeast extract, previously autoclaved at 120 C during 15 minutes and pH 6. 5mm of fungi removed from colony edge was used to inoculate in submerged culture under agitation in Sheaker at 180 rpm and 28 C. High (2.5g) and low (25mg) dosage of seryngaldazine was used. After three days of cultivation, the media was filtrated and enzymatic activity determined by XVII spectrophotometer. P. sanguineus had higher micelial growth and FSF11lower at concentration of 120 mcg of biotin. On the whole, biotin in the medium contribute to add micelial growth. Laccase increase when use higt concentration of inductor seryngaldazine. Laccase activity to P. sanguineus increase significantly when higher dosage of seryngaldazine at 3:5 glucose/ yeast extract was used. Hence it follows that vitamin, as for example biotin, contribute to micelial growth more rapid and higher as well as a major concentration of inductor (seryngaldazine) contribute to increase laccase activity.
Seleção de fungos isolados de plantas da Região Amazônica como fornecedores de lipases para biotransformações
(Universidade do Estado do Amazonas, 2006-12-15) Lisbôa, Líliam Ururahy de Souza; Zanotto, Sandra Patricia; Zanotto, Sandra Patricia; Pereira , José Odair; Marques, Alberto dos Santos
In this work 95 fungi of the Amazon region have been studied: endophytic and phytopathogenic isolated from plants, and wood degrading. When submitted to the assay with tributyrin, 79 isolates presented indicative hydrolytic activity. Of the 79 isolates, 26 that showed better performances of growth in liquid cultures have been submitted to the biocatalytic evaluation. Among the 26 mycelium-bound lipases that were evaluated as catalysts of esterification of oleic acid with n-pentanol in hexane, isolated UEA_027 presented conversions of 72% in 72 hours of reaction. The lipase enantioselective capacity of the isolated UEA_027 was verified through the resolution of (R,S)-2-hexanol and of (R,S)2-octanol by transesterification with vinyl acetate and hexane as solvent. The selected fungus resolved the alcohol (R,S)-2-octanol with enantiomeric excess of product above 88% and enantiomeric ratio above 20. Key words: Fungi, lipase, biotransformation, enantioselectivity, amazon region.
Ação Larvicida de espécies da família piperaceae em anopheles sp e Culex quinquefasciatus (DIPTERA: CULICIDAE)
(Universidade do Estado do Amazonas, 2007-04-23) Corrêa, Rachel Geber; Tadei, Wanderli Pedro; Nunomura, Sergio Massayoshi; Tadei, Wanderli Pedro
Extracts from eight species of the Piperaceae family were utilized to evaluate the larvicidal activity against 3rd instar larvae 3rd of Culex quinquefasciatus e Anopheles sp. Plant material that belongs to the Piper genera were collected from two places: Piper hostmanianum, P. dilatatum e P. tuberculatum were collected at the main campus of INPA – Instituto Nacional de Pesquisas da Amazônia, in Manaus, capital of the Amazonas State. The remaining species P. amapense, P. cyrtopodon, P. capitarianum, P. baccans e P. erectipillum were collected at the Adolpho Ducke reserve located at peripheric area of Manaus. The extracts were obtained from a standardized procedure that involved the extraction of the dried material in a Soxhlet apparatus for 18 h with methanol replacing the solvent every 6 hours. The evaluated extracts were obtained from different parts of plants – fruits, leaves, stems and roots. The biologic tests were made in rejoinders of 3 inside dismissable 50 mL cups containing 10 mL of distilled water within variable concentrations of 0,008 to 1000 μg/mL by making the mortality reading in 24 h. The most active extract was fractioned by the means of chromatographic techniques and the biomonitoring test enabled to isolate the main pelitorina e piplartina components that were identified by the means of monodimensioned nuclear magnetic resonance (RMN 1H e 13C). The studies of the larvicidal activity with the fractions and the isolated substance indicated a sinergystic effect among them. There were also registered external and behavior morphologic alterations of the 3rd instar larvae of A. nuneztovari in contact with the hexanic and chloroformic fruit extracts of Piper tuberculatum.
Estudo da Variabilidade Genética em Populações de Aedes albopictus (Diptera: Culicidae) no Estado do Amazonas, por meio de RAPD-PCR
(Universidade do Estado do Amazonas, 2007-05-22) Cáuper, Fábio Raphael Moreira; Santos, Joselita Maria Mendes dos; Santos, Joselita Maria Mendes dos
It has been used the RAPD/PCR technics to analysis the genetic Variability in Aedes albopictus, from Cidade Nova, Colina do Aleixo and Compensa neighborhoods of the city of Manaus and Tabatinga municipality in Amazonas state. The results of the RAPD profile analysis revealed 86 bands (locos) in the four populations with variations between 320 and 2.100 pb. Among the four analized populations, the n from Tabatinga has showed greater genetic variability. Even though by the polymorphic locus percentage (P = 100,00) or by its heterozigosity (Ho = 0,3929; He = 0,3981). The smallest genetic variability was detected at the Compensa population (P = 91,83; Ho = 0,3122). Of the three Manaus populations the one from Cidade Nova has shown the greatest genetic variability (P = 97,95; Ho = 0,3662) The q-square test for population differentiation has been significative (2= 466,7269; GL= 98; P < 0,000). The genetic structure was tested being its average value of Fst = 0,0662 ± 0,0096, showing a low genetic structure. Even though, this result has shown that genetic flux is happening among populations and the crossing is much probably aleatory. The values of genetic distances were very low (D= 0,0156 – 0,0786), showing that, populations are genetically very close, being Tabatinga and Compensa, the shortest genetic distance was detected among Cidade Nova and Colina do Aleixo populations (0,0786). The high polymorphism observed on the four analyzed populations cold be a suspicion of a greater genetic plasticity, occurring from its own genetic structure as an adaptable strategy to the Amazonian environment.
Efeito da adubação na nutrição e na colonização radicular por fungos micorrízicos arbusculares do camucamuzeiro (Myrciaria dubia (H.B.K.) McVaugh e da aceroleira (Malphigia punicifolia L.) em um Latossolo da Amazônia Central
(Universidade do Estado do Amazonas, 2007-05-24) Bezerra, Thana Esashika; Oliveira, Luiz Antonio de; Oliveira, Luiz Antonio de; Cavallazzi, José Renato Pereira; Oliveira, Arlem Nascimento de
The low fertility of the majority of Amazonian uplands soils allied to the deficiency resource of the majority of small farmers influence directly in the development and local agriculture sustainability. One strategies to reach the sustainability is to maximize the beneficial microorganisms use of the soil, as the arbuscular mycorrhizal fungi (AMF), which are capable to increase the site mineral nutrients and water absorption to host plants. In addition, the foliar fertilization is a economic practice that has high index of mineral nutrients use for the plants, due to lower amounts of the product used. In this manner, informations about soil microbiology and the acerola and camu-camu behavior in differents treatments can contribute to develop the production of these two potencial fruit species to local agriculture. This research had as objective to evaluate the fertilization effect in the nutrition and AMF colonization of the camu-camu and acerola tree cultivated on yellow oxisol located in the Brasileirinho Agricultural Community, Manaus. In fields conditions, it was evaluated the foliar macro e micronutrients contents, the mycorrhiza colonization rates and the spores number of both the species on three collect times. The experimental design was randomized blocks with a 3 x 10 split plot scheme to camu-camu tree. The factors means three collect times (08/04/06, 15/07/06 e 07/11/06) and ten treatments with chemical, foliar and organic fertilization. To acerola tree, 3 x 9 split plot scheme, with three collect times (21/02/06, 15/06/06 e 09/11/06) and nine treatments. There were significant variations in the acerola foliar contents in differents treatments. The plants that received the treatments with chemical fertilization presented the higher leand Ca, Mg, K, P and N. The plants that received the treatments with foliar fertilization presented the highest Fe, Mn and Zn leand contents. This fact provides the aiding on micronutrients absorption through the highest effectiveness of foliar fertilizer. There were variations in foliares contents of camu-camu in differents treatments, with exception of the N and Mn contents. The plants that received the treatments with organic fertilization presented the highest Ca, Mg, P and K averages. The Fe, Mn and Zn leand contents were favored by foliar fertilizer applications, exception to Mn, that did not present significant variations. In Authoress: THANA ESASHIKA BEZERRA Adviser: Dr. LUIZ ANTONIO DE OLIVEIRA acerola, the treatment with chemical fertilizer provided the hightest Ca, Mg, K, P and N contents without differing of the contents provided by organic fertilization. The organic fertilization is more economical than others fertilization to the small farmers. The foliar fertilizer was efficient to micronutrients, proving to be a quick, economical and of easy application. In camu-camu, the treatment with organic fertilization was adjusted, because it had provided the highest Ca, Mg and K contents. To P, Fe, Zn and Mn nutrients, the treatments with foliar fertilizer showed efficient. On acerola, the association with mycorrhiza fungi showed positives correlations with leand nutrients, but the correlation rate was very low. Thus, there was no efficient benefice of the plant-fungi symbiosis. In both the cultures, the fertilization and the P concentration in the soil can have influenced, of negative form, in roots colonization by mycorrhizal fungi. Besides, to camu-camu and acerola tree, the organic fertilization, generally, provided the better results to the macronutrients, and the foliar fertilization to the micronutrients. Key words: Amazonian Fruit Species, Foliar Fertilization, Plant Nutrition, Arbuscular Mycorrhizal Fungi (AMF).
Análise da Sobrevivência e Desenvolvimento de Mudas de Pau-Rosa (Aniba rosaeodora Ducke) em Clareiras Artificiais
(Universidade do Estado do Amazonas, 2007-05-25) Fonseca, Gisele Cândida Freitas da; Sampaio, Paulo de Tarso B.; Ferraz, João Baptista S.; Sampaio, Paulo de Tarso B.
In different sizes of artificial gaps, located at Estação Experimental de Silvicultura Tropical (EEST-INPA), in Manaus - AM, survival, development, herbivory susceptibility related to density and the nutritional estate of rose-wood plants (Aniba rosaeodora Ducke) were analyzed after three years of its establishment. The experiment was realized in three different size gaps (little gap, median gap and big gap), and a control-area under closed tree top was used to compare the influence of different luminosity incidence on plant growth. The gaps and the control-area are the different treatments. The data taken was: survival, by counting the number of plants, stem height, diameter, estimated herbivory percentage to calculate the herbivory index and determination of the nutritional state by chemical laboratorial analysis of material leaves. These data were taken in March and November of the year 2005, and analyzed by ANOVA and Tukey (5% significance) tests. Better plant development was observed in a bigger gap. It was not observed differences of survival between the treatments. There were no evidences of herbivory related to density. It was observed nutritional deficit in all treatments.
Estudo de marcadores químicos de guaraná (Paullinia cupana) por CLAE
(Universidade do Estado do Amazonas, 2007-06-22) Stark, Gisele; Nunomura, Sergio Massayoshi; Nunomura, Rita de Cássia Saraiva; Nunomura, Sergio Massayoshi
Guaraná, Paullinia cupana var. sorbilis (Mart.) Ducke, is one of the most important natural products of the Amazon basin and very appreciated abroad. The Indians of low Amazon due to its stimulating, fortifying and anti-diarrhea properties consume it and today guarana is also cultivated in other states of Brazil. Its seeds, powdered or as syrup, are used in many products (beverages). The main chemical constituents of guarana are methylxanthines (caffeine, theofiline, theobromine) and also phenolic compounds (catechins). Due to the increasing interest for guarana, adulteration based on synthetic caffeine addition (low cost) has been more often found. This adulteration cannot be detected by the main quality control methods based on the quantification of caffeine. This work describes the development of an analytical method developed by reversed phase high performance liquid chromatography to acquire chromatographic profiles (fingerprints) of guarana to quality control and assess adulteration of guarana based on the analysis of its multicomponents. The fingerprints were very reproductible, relative to five chemical markers (caffeine, catechin, epicatechin, theofiline and theobromine). The method was also able to quantify caffeine in the samples. Twenty samples from three different states in Brazil (Amazonas, Bahia and Mato Grosso) were analysed and a common fingerprint, independent from its origin, was obtained. The caffeine content of the samples could assess the geographical origin. The certification of origin of a natural product is one of the most important added values. The antioxidant power of powdered guarana seeds obtained in two different ways (toasted or sun dried) were also evaluated and compared to other plant species consumed worldwide as beverages and recognized for their antioxidant power (green tea, black tea, coffee, mate and barley). The sample of toasted guarana had a significant antioxidant power, inferior only to green tea, whereas the sun dried sample have much less activity. This indicates the great potential of guarana and the importance of the drying process of the seeds to keep its nutraceutical properties. Key-words: Chromatographic profile, antioxidant activity, HPLC, quality control, phenolic, methylxanthines.
Análise da variabilidade genética em populações de Anopheles darlingi Root, 1926 (Diptera: Culicidae) do Estado do Amazonas, usando marcadores RAPD
(Universidade do Estado do Amazonas, 2007-07-15) Silva, Ana Paula Barbosa da; Santos, Joselita Maria Mendes dos; Santos, Joselita Maria Mendes dos
Anopheles darlingi populations from Manaus, Coari, São Gabriel da Cachoeira and Tabatinga were analyzed by using the RAPD molecular marker, for the purpose of assessing the genetic variability and differentiation between them. Populations captured indoor and outdoor (around the house, corral) were also analyzed in Coari and Manaus, between 17:00h and 05:00h, in January and February of 2006. Genetic variability findings were high for mosquito’s four populations, showing it to be higher in São Gabriel da Cachoeira (P= 97.37%; He= 0.3202), and lower in Manaus (P= 78.94%; He= 0.2741). The chi-square test was significant (c2= 1589.5700; GL= 304; P < 0.001). The genetic structure analysis showed significant FST value (FST= 0,0851 ± 0,0075), indicating a reduced gene flow between populations. The “bootstrapping” consistency index for 1000 replicates was 95%. The genetic distance between populations was low (D= 0.0095 – 0.0502), showing to be lower between Coari and Tabatinga, and higher between São Gabriel da Cachoeira e Tabatinga. Despite the latter presenting higher genetic and geographic distance, data showed no clear relation with the Isolation by Distance Model (IBD), since Coari and Tabatinga revealed higher genetic similarity; however the lower geographic distance was between Manaus and Coari. Genetic variability analysis based on mosquito biting activity patterns, showed mosquitoes captured indoors presented highest genetic variability, where the polymorphism and expected heterozygosity were higher in both populations (Coari: P= 84.86% e He= 0.3069; Manaus: P= 78.94% e He= 0.2741). The chi-square test for the parameters was significant (c2= 695.8958; GL= 304; P < 0.001). Genetic structure analysis also showed a significant FST value (FST= 0.0775 ± 0.0072). The genetic distance between populations was low. For Manaus, the genetic distance between indoor and outdoor sub-populations was 0.0004, there being certain homogeneity among them. For Coari, the genetic distances were slightly higher, the highest being that between the outdoor (cattle) and the indoor (D= 0.0296), and the lowest between the indoor and the outdoor (D= 0.0081). As a whole, the data showed high genetic similarity between the analyzed populations, despite the little genetic structuring found. The highest genetic variability found the indoor, in the Coari and Manaus populations, indicates a higher genetic plasticity, and so it might confer them highest adaptability to the changes occurring in the environment.
Desenvolvimento e validação de método para quantificação da capacidade redutora de extratos vegetais e secos
(Universidade do Estado do Amazonas, 2008-01-22) Novaes, Jussival de Abreu Pinheiro; Nunez, Cecilia Veronica; Nunez, Cecilia Veronica; López-Lozano, Jorge Luis; Sargentini Junior, Ézio
The developed method in this work is defined as a chemical method of indirect quantification of the reducing capacity of vegetable extracts. It is based on the reduction of ions Fe3+ by chemical substances contained in the extracts and subsequent quantification of Fe2+ ions for spectrophotometric technique in range of the visible using the adapted method of the reagent 1,10-phenanthroline. The procedure of the development and validation was composed by three parts: characterization of the quantification of Fe2+ in the form of the complex [Fe(fen)3]2+; reduction of ions Fe3+ by organic compounds and vegetable extracts; and characterization of the proposed method. Ascorbic acid, hydroquinone, catechol, pirogallol, quercetine, Trolox™, BHA, BHT, cisteine and glutathione were used as reference reducing. The samples of dry vegetable extract were obtained from bank of extracts of the Laboratory of Bioprospection and Biotechnology, installed in the Coordenação de Pesquisas de Produtos Naturais (CPPN) of the Instituto Nacional de Pesquisas da Amazônia (INPA). The criterion for the choice of the extracts used in the validation process was based on measures (peaks or oxidation bands) by cyclic voltammetry technique. The compound [Fe(fen)3]2+ is stable and in the tested conditions it did present maximum absorption in 508.8 nm with molar absorptivity around (1.084 ± 0.003) × 104 L.mol-1.cm-1. The quantification of ions Fe2+ in the form of the compound [Fe(fen)3)]2+ is linear in the range from 0.2 to 10.0 g.mL-1. The adjusted line obtained by the method of minimum squares presented equation ABS = 0.01444 ± 0.19417 × [Fe2+] (R2 = 0.9994) for concentrations up to 10.0 g.mL-1 and ABS = 0.00263 ± 0.19974 × [Fe2+] (R2 = 0.9997) up to 5.5 g.mL-1. Established oxidation process consists of two stages: (1) reaction with ions Fe3+ and (2) reaction with ions Fe3+ in the presence of 1,10-phenanthroline. The oxidation degree and the linearity vary in agreement with the reaction mechanism. The proposed method presented positive answer for the glutathione, differently of other methods described in the literature. The developed method and validated presents the following characteristics: Limit of decision, of detection and of quantification was of 0.17 g Fe2+.mL-1 , 0.20 g Fe2+.mL-1 and 0.31 g Fe2+.mL-1, respectively; Reproducibility and repeatability between 0.5% and 3.9%; absolute discrepancy between 1.3% and 3.9% for the following reducing: ascorbic acid, hydroquinone and BHA. Ethanol, methanol, acetone and DMSO can be used to dissolve the vegetable extracts. Ethylene glycol and DMF can be used, but they can coordinate with Fe2+/Fe3+ ions. The coordination of the Fe2+/Fe3+ ions for ligands present in the extract are labile in relation to chelante 1,10-phenanthroline. The Co2+, Cu2+, Ni2+, Mn2+, Sn2+, Zn2+, Mg2+, Ca2+, Na+, K+, NH4+, SO42-, Cl- e NO3don't interfere in the quantification of the complex [Fe(fen)3]2+, but PO43- ion interfere in higher concentrations than 10 g Fe2+.mL-1. The UV radiation produces false positive, converting Fe3+ in Fe2+. With base in the obtained results, it can be concluded that the method developed for quantification of the reducing capacity of vegetable extracts was appropriate and of easy execution.