Isolamento de uma quitinase extraída do cefalotórax do camarão marinho Litopenaeus schmitti (BURKENROAD, 1936) : avaliação de suas atividades antimicrobiana e larvicida

Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.

Polissacarídeos sulfatados de interesse farmacológico no camarão litopenaeus schimitti

Sulfated Polysaccharides with unique chemical structures and important biological activities has been found in a diversity of sea invertebrates. For that, to exist a huger interest on the biotechnology field in the research theses sulfated compounds isolated from sea organisms. Despite the privileged brazilian position for these compounds attainment, there are still a few scientific informations about the isolated substances and their biological activities. A head the displayed, the present work has for objectives, to evaluate the pharmacological properties of the glycosaminoglycans isolated from the sea shrimp Litopenaeus schimitti on homeostasis, blood coagulation, leukocytes migration and platelet/leukocyte adhesion. For this, yhe glycosaminoglycans were extracted from crustacean tissues by proteolysis, fractionation with acetone and later submitted to pharmacological assays. The crustacean tissues showed compounds heparin-like, with anticoagulant activity of 45 IU/mg and 90 IU/mg, respectively. These molecules showed low residual hemorrhagic effects in the tested concentration (100 µg/mL), when compared to unfractionated commercial heparin (UFH). Another dermatan sulfate-like compound, predominately constituted for disulfated disaccharides, was isolated from crustacean abdomen. This compound showed an efficient effect on leukocytes migration inhibition, in the concentration of 15 µg/mL, reducing the cellular infiltration in 65% when compared to the controlled animals. In this same concentration, the DS reduced in 60% the protein concentration of the peritoneal exudates. In the concentration, this compound of 0.5 mg/mL, it was capable to reduce in 40% platelet/leukocytes adhesion. Our data demonstrate that these sulfated polysaccharides isolated from the shrimp L. schimitti will can be used as bioactive compounds, appearing as active principles for pharmacological development, anticoagulants and inflammatory response regulators

Avaliação do potencial anti-inflamatório de composto tipo heparina (cCTH) extraído do caranguejo Goniopsis cruentata em modelo experimental de peritonite

Heparin, a sulfated polysaccharide, was the first compound used as an anticoagulant and antithrombotic agent. Due to their structural characteristics, also has great potential anti-inflammatory, though such use is limited in inflammation because of their marked effects on coagulation. The occurrence of heparin-like compounds that exhibit anticoagulant activity decreased in aquatic invertebrates, such as crab Goniopsis cruentata, sparked interest for the study of such compounds as anti-inflammatory drugs. Therefore, the objective of this study was to evaluate the potential modulator of heparin-like compound extracted from Goniopsis cruentata in inflammatory events, coagulation, and to evaluate some aspects of its structure. The heparin-type compound had a high degree of N-sulphation in its structure, being able to reduce leukocyte migration into the peritoneal cavity at lower doses compared to heparin and diclofenac sodium (anti-inflammatory commercial). Furthermore, it was also able to inhibit the production of nitric oxide and tumor necrosis factor alpha by activated macrophages, inhibited the activation of the enzyme neutrophil elastase in low concentrations and showed a lower anticoagulant effect in high doses as compared to porcine mucosal heparin. Because of these observations, the compound extracted from crab Goniopsis cruentata can be used as a structural model for future anti-inflammatory agents

Atividades antinociceptiva e anti-inflamatória de uma fração rica em heterogalactana sulfatada extraída da alga codium isthmocladum (vickers 1905)

Sulfated polysaccharides comprise a complex group of macromolecules with a range of several biological activities, including antiviral activity, anticoagulant, antiproliferative, antiherpética, antitumor, anti-inflammatory and antioxidant. These anionic polymers are widely distributed in tissues of vertebrates, invertebrates and algae. Seaweeds are the most abundant sources of sulfated polysaccharides in nature. The green algal sulfated polysaccharides are homo or heteropolysaccharides comprised of galactose, glucose, arabinose and/or glucuronic acid. They are described as anticoagulant, anti-inflammatory, antiviral, anti-angiogenic, antitumor compounds. However, there are few studies about elucidation and evaluation of biological/pharmacological effects of sulfated polysaccharides obtained from green algae, for example, there is only one paper reporting the antinociceptive activity of sulfated polysaccharides of these algae. Therefore this study aimed to obtain sulfated polysaccharides of green seaweed Codium isthmocladum and evaluates them as potential antinociceptive agents. Thus, in this study, the total extract of polysaccharides of green alga C. isthmocladum was obtained by proteolytic digestion, followed by fractionation resulting in five fractions (F0.3, F0.5, F0.7, F0.9 and F1.2) by sequential precipitation with acetone. Using the test of abdominal contractions we observed that the fraction F0.9 was the most potent antinociceptive aompound. F0.9 consists mainly of a sulfated heterogalactana. More specific tests showed that Fo.9 effect is dose and time dependent, reaching a maximum at 90 after administration (10 mg / kg of animal). F0.9 is associated with TRPV1 and TRPA1 receptors and inhibits painful sensation in animals. Furthermore, F0.9 inhibits the migration of lymphocytes induced peritonitis test. On the other hand, stimulates the release of NO and TNF-α. These results suggest that F0.9 has the potential to be used as a source of sulfated galactan antinociceptive and anti-inflammatory

Estrutura da população de Coleodactylus natalensis Freire, 1999 (Squamata: Sphaerodactylidae) no Parque Estadual Dunas de Natal, Rio Grande do Norte, Brasil

Coleodactylus natalensis Freire, 1999, an endemic species of Atlantic Forest fragments around the Natal municipality, Rio Grande do Norte State, Brazil, has as type locality the Parque Estadual Dunas de Natal (05º48 S to 05º53 S and 35º09 W to 35º12 W), one of the largest restinga (herb and shrub association on sand dunes along the Brazilian coastline) associate fragment, surrounded by urban zone, placed on setentrional Atlantic Forest limits. We made estimates on populational density, spatial distribution, habitat and microhabitat preferences and feeding ecological aspects like sazonal and sexual variations on diet, prey electivities and niche breadth. We randomly sampled ninety-six 50m2 quadrants in each of the four habitats identified in the study area. Were collected 49 specimens and their stomach contents were analyzed; prey items found were correlated with leaf-litter invertebrates from habitat samples. We found a 98,5 ± 75,5 individuals/ha density, in grouped distribution pattern on densest habitats and random distribution on others habitats. This species lives mostly on leaf-litter in forest habitats, in higher humidity points, with lower temperatures, deeper leaf litter and lower sea level elevations than the randomly chosen points in the study area. Isopoda and Aranae were the most important prey categories in numeric, frequency and volumetric terms. Niche breadth has an intermediate value and was variable in sexual and in habitat terms. There was no correlation between morfometric measures and prey size on diet. The C. natalensis population studied seems to be diet opportunist, although selects larger prey items. The Parque Estadual das Dunas do Natal has several indications of anthropic pressure from the surrounding urban area that may affects the local C. natalensis population. Thus, the fragility of this species calls for urgent conservation efforts

Ecologia alimentar de aves insetívoras de um fragmento de mata decídua do extremo norte da Mata Atlântica

The diet study of birds has contributed historically as a model for use to understanding ecological patterns and strategies used by several other groups of vertebrates, which are observed in season patterns and temporal availability of resources, and other. This study has as objective generate information concerning the diet of insectivorous birds during rainy season and dry season, as well as analyze Index food importance, niche overlap, niche breadth, electivity, and seasonal availability of prey. The study was conducted in a fragment of about 270 ha (center coordinates and 5 º 53'S 35 ° 23'W). The sampling of birds occurred between March 2008 and December 2009 in three pre-established trails. Catches of birds were performed using 10 mist nets placed in line, where each trails was sampled once a month. Samples of pellets were obtained by means of tartar emetic. Sampling of availability of prey occurred between February 2009 to December 2009. We used two methods of sampling (pitfall traps and Shake cloths). We captured 269 individuals of 21 species of insectivorous birds. We collected 4116 invertebrates of which 3259 in the rainy season and 857 in the dry season. We obtained 174 samples stomach, where 10 species were exclusively insectivorous diet, nine fed on insect/plant material, an insect/plant material/vertebrate and one for insect/vertebrate. During the rainy season was observing difference between the consumption of items with higher food importance. The Coleoptera was item with higher food importance (73%), followed by Formicidae (7%) and Araneae (6%). During the dry season, no difference was found difference between the consumption of items with higher food importance. The Coleoptera was item with higher food importance (34%), followed by seeds (29%) and Formicidae (18%). The highest levels of niche overlap occurred during the rainy season, while the dry season was characterized by high levels of niche 11 segregation. This indicates that the local insectivorous birds community was structured differently between periods. No was found correlation between the values of niche breadth to the mean weight of the body size. We observed seasonal patterns in prey availability, with the peak availability of invertebrates observed seasonal patterns in rainy season. The insectivorous birds selected the same species richness during both periods, showing a specialized diet. Thamnophilus pelzelni was the only species that had their diet influenced by seasonality. Regarding the overall diet of insectivorous birds, observed a high consumption of prey, whose food availability caused the birds could invest and increase their food resources

Nuclear-follower foraging associations among Characiformes fishes and Potamotrygonidae rays in clean waters environments of Teles Pires and Xingu rivers basins, Midwest Brazil

Por meio de observações sub e supra-aquáticas foram registradas associações alimentares do tipo nuclear-seguidor entre três espécies de peixes characiformes - Chalceus epakros, Hemiodus semitaeniatus e Hemiodus unimaculatus - e uma espécie de raia de água doce - Potamotrygon orbignyi - nas bacias dos rios Teles Pires e Xingu, no Centro-Oeste do Brasil. Os peixes teleósteos foram observados seguindo as raias quando estas revolviam o substrato à procura de invertebrados, formando discretas nuvens de sedimento. Essas situações atraíram os peixes que se aproximaram das raias para se alimentar de pequenas presas e outros tipos de alimentos expostos desta forma. Esse é um típico exemplo de relação comensal onde um participante é beneficiado enquanto o outro não é prejudicado e representa o segundo registro na literatura de associação alimentar do tipo nuclear-seguidor entre raias potamotrigonídeas e peixes teleósteos, demonstrando o potencial de estudos naturalísticos para a descoberta de novas interações envolvendo espécies de peixes de água doce.

Soroprevalência da infecção pelo Trypanosoma cruzi no município de Caicó, RN, e caracterização genética por RAPD do parasito isolado de humanos e de triatomíneos no semiárido potiguar

Trypanosoma cruzi infection was evaluated in 390 resident individuals in different rural communities of Caicó municipality, State of Rio Grande do Norte (RN). Of 28 investigated communities the soroprevalence of T. cruzi infection was 2.8% in eight rural communities individuals. The epidemiological characteristics of seropositive shown that the age ranged from 22 to 64 years, being significantly raised from 31 years (90.9%). The female gender was predominant and low education degree. Those individuals reported that they never donated blood, but they had direct contact with triatomines bug. The isolation of the parasite was performed by blood culture and xenoculture methods to determine the genetic variability of the samples. Twenty seven T. cruzi isolates were analyzed by RAPD as genetic marker using three random primers (M13-40, gt11-F and L15996). The T. cruzi isolates showed 73.7% of shared bands considering the average obtained with the three primers, and were genetically well correlated. Using this marker it was possible to separate the populations of the parasite in three distinct groups. The first group composed by isolates obtained of triatomines and humans from four different districts (Caicó, Caraúbas, Serra Negra doNorte and Governador Dix-Sept Rosado); the second contained isolates obtained of triatomines of two different species (T. brasiliensis and P. lutzi) captured in Caraúbas and Serra Negra do Norte. The third grouped isolates obtained from humans of Angicos and Caicó municipalities. In different localities of distinct mesoregions, State of RN, a profile genetic well correlated was identified among all isolates and the presence of three distinct groups of the parasite circulating among vertebrate and invertebrate hosts

Efeitos do plantio direto e da consorciação soja-milho sobre inimigos naturais e pragas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversidade e distribuição espacial de artrópodes associados ao solo em agroecossistemas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Relações ecologicas entre caranguejos e composição arbórea em bosques de mangue naturais e restaurados no nordeste brasileiro

This study aimed to compare the development of crab and tree communities of two restored mangrove areas, one planted with Rhizophora mangle and the other naturally recovered, and also to compare the predation of Grapsid crab Goniopsis cruentata and the Ocypodid Ucides cordatus over the propagules of three mangrove trees: Rhizophora mangle, Avicennia schaueriana e Laguncularia racemosa. Specifically, we tested the hypothesis that Goniopsis predation is more important that Ucides predation, and that these consumers have antagonist effects over propagule consumption. In each area, 10 quadrates were selected at random to analyze tree richness, diameter, height, tree biomass and crab richness and density five years after restoration experiment start. Results show that tree height, biomass and crab density were significantly higher in artificially restored area. No significant differences were observed in crab species richness between areas, but higher tree richness was observed in self-recovered area. Results suggest that planting propagules of Rhizophora can significantly increase tree recovering if the aim was increase tree biomass and crab density, which can accelerate return of ecological functionality. Goniopsis is a more important propagule predator than Ucides both in natural and restored areas. The effects of Goniopis were higher in absence of Ucides, due to negative interactions among these two predator species. The preference of Goniopsis by Avicennia and Laguncularia can favor the dominance of Rhizophora observed in Neotropical mangroves. This study suggests that propagule predation by Goniopsis should be controlled in restoration programs, if dominance of Rhizophora is undesirable respect to more rich tree communities

Análise de fauna e flutuação populacional de Carabidae e Staphylinidae (Coleoptera) em sistemas de plantio direto e convencional

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aspectos Biológicos dos Estádios Imaturos de Chrysoperla externa (Hagen) e Ceraeochrysa cincta (Schneider) (Neuroptera: Chrysopidae) Alimentados com Ovos e Ninfas de Bemisia tabaci (Gennadius) Biótipo B (Hemiptera: Aleyrodidae)

Foram avaliados os aspectos biológicos dos estádios imaturos de Chrysoperla externa (Hagen) e Ceraeochrysa cincta (Schneider), alimentados com ovos e ninfas de Bemisia tabaci (Genn.) biótipo B. Ovos dos predadores foram individualizados em recipientes de vidro de fundo chato com 2,5 cm de diâmetro e 8,5 cm de altura que foram mantidos à temperatura de 25±2ºC, UR de 70±10% e fotofase de 14h. Após a eclosão das larvas, discos de folhas de tomateiro e de couve com 2 cm de diâmetro, contendo ovos e ninfas de B. tabaci, respectivamente, foram colocados nos recipientes. A duração e a viabilidade de cada ínstar e da fase pupal de ambas as espécies predadoras alimentadas com diferentes estágios da presa foram avaliados. Larvas de primeiro ínstar de C. externa apresentaram desenvolvimento mais lento quando alimentadas com ovos de B. tabaci biotipo B monstrando influência do estágio da presa fornecida no período de desenvolvimento do predador. O mesmo não ocorreu para a espécie C. cincta. O desenvolvimento de larvas de primeiro ínstar de ambas as espécies predadoras foi semelhante quando foram alimentadas com ovos de B. tabaci; porém C. cincta apresentou período de desenvolvimento mais longo (5,4 dias) comparada a C. externa (4,0 dias) quando alimentada com ninfas da presa. Para larvas de segundo e terceiro ínstares e para a duração da fase larval não houve influência do estágio da presa fornecida (ovo ou ninfa) no período de desenvolvimento. A duração do segundo ínstar foi maior para C. cincta do que para C. externa (4,8 e 4,1 dias, respectivamente). Porém a duração do terceiro instar foi maior e da fase larval foi menor para C. externa (6,6 e 15,3 dias, respectivamente) do que para C. cincta (6,4 e 16,7 dias, respectivamente). A viabilidade dos ínstares foi acima de 90% para ambas as espécies. No entanto, menor porcentagem de viabilidade da fase de pupa foi registrada para C. cincta, quando alimentada com ovos (30%) ou ninfas (55,6%).

Mite diversity on plants of different families found in the Brazilian Atlantic Forest

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Potencial reprodutivo de Chrysoperla externa (Hagen) (Neuroptera, Chrysopidae) em função do número de indivíduos por unidade de criação

Verificou-se o efeito de diferentes densidades de insetos por unidade de criação sobre o potencial reprodutivo de Chrysoperla externa. Utilizaram-se adultos da geração F4 mantidos a 25 ± 1ºC, 70 ± 10% de UR e fotofase de 12 horas. Foram utilizadas as seguintes densidades por unidade de criação (10 cm de diâmetro por 23 cm de altura e volume de 1650 cm³): um macho e três fêmeas, dois machos e seis fêmeas, três machos e nove fêmeas e quatro machos e doze fêmeas. Avaliou-se o período de pré-oviposição, a oviposição diária e total por fêmea e por unidade de criação, as porcentagens de ovos viáveis e inférteis e o índice de aproveitamento, calculado por fêmea e por unidade de criação. A densidade quatro machos e doze fêmeas apresentou melhor aproveitamento da unidade de criação, compensando as reduções observadas nos valores de postura média e total por fêmea.