Atividade β-D-N-Acetilglucosaminidásica de enzimas imobilizadas extraídas da Artemia franciscana e possíveis aplicações biotecnológicas

A β-D-N-acetilglucosaminidase extracted and partially isolated from crustacean Artemia franciscana by ammonium sulfate precipitation and filtration gel chromatography Bio Gel A 1.5m. the enzyme was immobilized on ferromagnetic Dacron yielding a insoluble active derivative with 5.0 units/mg protein and 10.35% of the soluble enzyme activity. β-D-N-acetilglucosaminidase-ferromagnetic Dacron was easily removed from the reaction mixture by a magnetic field, it was reused for ten times without loss in its activity. The ferromagnetic Dacron was better activated at pH 5.0. The particles visualized at scanning electron microscope (SEM) had presented different sizes, varying between 721nm and 100µm. Infra red confirmed immobilization on support, as showed by primary amino peaks at 1640 and 1560 cm-1 . The immobilize enzyme presented Km of 2.32 ± 0.48 mM and optimum temperature of 50°C. Bought presented the same thermal stable of the soluble enzyme and larger enzymatic activity at pH 5.5. β-D-N-acetilglucosaminidase-Dacron ferromagnético showed sensible for some íons as the silver (AgNO3), with loss of activity. The β-D-N acetilglucosaminidase activity for mercury chloride (HgCl2), whom is one of the most toxic substance joined in nature, it was presented activity already diminished at 0,01mM and lost total activity at 4mM, indicating sensitivity for this type of metal. β-D-N-acetilglucosaminidase-ferromagnetic Dacron showed degradative capacity on heparan sulfate, the enzyme still demonstrated degradative capacity on heparan sulphate, suggesting a possible application to produce fractions of this glycosaminoglycan

Avaliação do potencial antiinflamatório, anticoagulante e hemorrágico de heparinóides presentes em "Artemia franciscana"

Heparan sulfate (HS) and Heparin (Hep) glycosaminoglycans (GAGs) are heterogeneous and highly charged polysaccharides. HS is structurally related to Hep but is much less substituted with sulfo groups than heparin and has a more varied structure (or sequence). Because of structural similiarities between these two polymers, they have been described together as heparinoids . Both chains bind a variety of proteins and mediate various physiologically important processes including, blood coagulation, cell adhesion and growth factor regulation. Heparinoids with structural characteristics similar to these described from HS and/or Hep from mammalian tissues have been isolated from different species of invertebrates, although only a few heparinoids from unusual sources have been characterized. The present study describes the presence of unusual heparinoids population from Artemia franciscana, isolated after proteolysis and fractionation by ion exchange resin and named, F-3.0M. The study model in vivo were hemostasis (rat tail scarification) and inflamatoty activity. The tests in vitro were used for coagulations assays (PT and APTT). The analyse of the heparinoids eluted with 3,0M NaCl showed electrophoretic migration in different buffer systems a single band with a behaviour intermediate between those of mammalian HEP and HS. The main products obtained from Artemia heparinoids after enzymatic degradation with heparitinases I and II from F. heparinum were N-sulphated disaccharides (∆U-GlcNS,6S/ ∆U,2S-GlcNS and ∆U-GlcNS) and N-acetylated disaccharides (∆U, GlcNAc). This heparinoid had a lower hemorrhagic effect (400μg/ml) when compared to unfractiionated heparins(25μg/ml).The results also suggest a negligible APTT activity of this heparinoid (62.2s). No action was observed on PT indicating that F-3.0M haven t action on the extrinsic pathway. The results showed that the fraction F- 3.0M have inhibitory effect on migration of leukocytes, 64.5% in the concentration of 10 μg/ml (P<0.001). The search for new heparin and/or heparan sulphates analogs devoid of anticoagulant activity is an atractive alternative and may open up a wide variety of new therapeutic applications

The Predation of Artemia Nauplii by the Larvae of the Amazon River Prawn, Macrobrachium amazonicum (Heller, 1862), is Affected by Prey Density, Time of Day, and Ontogenetic Development

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ingestion rates of Artemia nauplii for different larval stages of Macrobrachium rosenbergii

The functional response between ingestion rate and food concentration was determined for each larval stage of Macrobrachium rosenbergii. Artemia franciscana nauplii were supplied at 2,4, 6, 8, 10 and 12 per milliliter. The nauplii were counted by sight using a Pasteur pipette and transferred to Petri dishes containing 40 ml of brackish water (12 parts per thousand) lying on the top of black plastic. One larva at each stage was individually placed into each Petri dish containing different food density. After 24 h, each larva was removed from the Petri dish and the leftover nauplii were counted. The amount consumed was determined by the difference between the initial and final number of nauplii. Ingestion rate (I) increased as food density (P) increased and was defined by the model I=I-m(1-e(-kP)). The results suggest four levels of ingestion during larval development. The first level includes stages II, III and IV, with average maximum consumption of about 40 nauplii/day; the second level includes stages V and VI, with consumption of approximately 55 nauplii/day; the third level includes stages VII and VIII, with consumption of 80-100 nauplii/day. The fourth level includes stages IX, X and XI, in which the high values for maximum ingestion (Im) exceed the load capacity of the medium. The low values for constant k (that may correspond to the adaptability of the food to prey characteristics, such as, size, mobility, etc.) obtained for stages IX, X and XI indicated that Artemia is not an adequate prey and there is necessity of a supplementary diet. The best relationship between predator and prey seemed to occur during stage IV Results obtained in the present work may subsidize future researches and serve as a guideline for practical considerations of feeding rates. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Co-feeding of live feed and inert diet from first-feeding affects Artemia lipid digestibility and retention in Senegalese sole (Solea senegalensis) larvae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação hemodinâmica e metabólica da cetamina e cetamina S(+) após a reposição volêmica com hidroxietilamido 130/0,4 e solução salina hipertônica 7,5%

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

efeito da aplicação de uma solução salina no crescimento e na composição química da atriplex nummularia

This study aimed to analyze the effect of a saline solution on growth and chemical composition of Atriplex nummularia, shrubby plant, absorbing salts used in the diet of animals and the management of water and saline soils. These plant seedlings were planted and grown in a reserved area at the Federal University of Rio Grande do Norte. The plantation was divided into two blocks, in which one of them was irrigated with saline solution with a concentration of 2840 mgL-1 of NaCl and the second group was irrigated with drinking water. After six months, the plants were collected, harvested and divided into three parts: leaf, thin and thick stem. Monthly, dimension measurements were carried out for cataloging the growth of Atriplex. Ion Chromatography (IC) and Optical Emission Spectroscopy Inductively Coupled Plasma (ICP-OES) were used to analyze the chemical composition of the partition plant parts. The results of these analyses revealed that an absorption process of anions and cations by Atriplex nummularia plant during its growth was achieved, in particular by a higher concentration of sodium and chloride ions. Scanning electron microscopy images showed and confirmed the presence of small crystals on the leaf surface. Electrical conductivity and pH measurements of the aerial parts of the plant were carried out and these results showed that the leaf is the plant part where there is a largest concentration of ions. In addition, measurements of specific surface were obtained from irrigated plants with saline solution, achieving higher surface area, in all cases. Plant dimensions obtained monthly showed that the plants irrigated with water grew 5% more than those plants irrigated with saline solution. Based on results obtained, Atriplex plant showed a higher potential to survive and adapt to environments (aquatic or geological) with high levels of salinity and this property can be used as a tool for removing salts/metals from industrial contaminated soils and effluents.

Análise morfológica da corrosão por pites em aço inoxidável austenítico AISI 310S submetido à exposição em névoa salina

O artigo teve como objetivo estudar a evolução do crescimento, distribuição e classificação dos pites em aços inoxidáveis austeníticos AISI 310S no estado como recebida e tratado termicamente, submetidos a diferentes tempos de exposição em meio salino. A aplicabilidade deste trabalho baseou-se no desenvolvimento de uma técnica para caracterização morfológica da corrosão localizada, associado com os aspectos de descrição de formas, tamanho, parâmetros específicos e populacionais. A metodologia consistiu nas seguintes etapas: preparação dos corpos de prova, testes de corrosão via névoa salina em diferentes condições, análise microestrutural, análise dos perfis dos pites, processamento digital e análise de imagens, visando caracterizar a distribuição, morfologia e o tamanho dos pites. Os resultados obtidos no processamento digital e análise de imagens dos perfis foram submetidos á análise estatística, utilizando à mediana como parâmetro de avaliação na liga como recebido e tratada. A liga como recebido, exibe a seguinte morfologia: pites hemisféricos> região de transição A> região de transição B> irregulares> cônicos. A quantidade de pites na liga tratada a cada tempo de exposição é: região de transição B> hemisféricos> região de transição A> cônicos> irregulares.

Pomada orgânica natural ou solução salina isotônica no tratamento de feridas limpas induzidas em ratos

Foram utilizados 40 ratos Wistar, machos, com peso inicial médio de 225 gramas, separados em dois grupos. Após serem anestesiados com éter, foi induzida uma ferida limpa na região torácica dorsal cranial, mediante a utilização de molde metálico de 2,0 x 1,5cm. No grupo I, foi utilizada pomada contendo óleo de fígado de bacalhau, extrato de confrei, extrato cítrico, óxido de zinco e veículo (Creamex®); no grupo II, a ferida foi apenas limpa com solução salina isotônica. Para a avaliação histológica, foram submetidos à eutanásia cinco animais de cada grupo no 3º, 7º, 14º e 21º dia de pós-operatório. O grupo tratado com a pomada orgânica apresentou padrão cicatricial de qualidade superior em relação ao grupo tratado com salina isotônica, representado por formação de fibras colágenas, neoformação de vasos e reepitelização completa da epiderme.

Efeitos cardiovascular e metabólico da reposição volêmica com solução salina hipertônica 7,5% associada ou não ao hidroxietilamido 130/0,4 em gatas com hipovolemia induzida

A hipovolemia é caracterizada por uma perda de fluido corpóreo, cursando com inadequado fluxo circulatório e consequentemente lesão tecidual. Neste trabalho, objetivou-se comparar a expansão volêmica resultante da administração de solução salina hipertônica (NaCl 7,5%), isolada ou em associação com hidroxietilamido 130/0,4 (HES 130/0,4), em gatas com hipovolemia induzida, sob anestesia geral inalatória com isofluorano. Foram utilizadas 12 gatas, sem raça definida, adultas, com massa corporal média de 3,07±0,56kg. Os animais foram anestesiados com isofluorano e, após a preparação cirúrgica, foram mantidos em 1CAM sob ventilação controlada. Após a estabilização do plano anestésico, foram avaliados os parâmetros basais. em ato contínuo, iniciou-se a fase de hipovolemia, por meio da retirada de 30ml kg-1 de sangue da artéria femoral. Após 60 minutos da estabilização do quadro de hipovolemia, foi realizada nova mensuração dos dados, alocando-se os animais aleatoriamente em dois grupos: GSH (grupo solução hipertônica, n=6), que receberam, na fase de expansão volêmica, NaCl 7,5% isolada, na dose de 4ml kg-1, e GSHC (grupo salina hipertônica associado ao coloide, n=6), que receberam NaCl 7,5%, na mesma dose citada, em associação com HES 130/0,4, na dose de 30ml kg-1. Após realização do tratamento, foram avaliados novamente os efeitos cardiovasculares e hemogasométricos por 120 minutos. As pressões arteriais média (PAM), sistólica (PAS) e diastólica (PAD) foram maiores logo após a expansão volêmica (T0) para o GSH. de T45 até T120, as PAM, PAS e PAD foram maiores para o GSHC, em comparação com o GSH. A pressão venosa central foi maior no GSHC até T60. Não foram observadas diferenças entre grupos para frequência cardíaca e respiratória, íons sódio e potássio, déficit de base, bicarbonato, saturação de oxigênio na hemoglobina, glicose, PaCO2, PaO2 e pH. Conclui-se que a administração de NaCl 7,5% isoladamente aumenta rapidamente a PAM, PAS e PAD em gatos com hipovolemia induzida, mantendo esse efeito por apenas 30 minutos, enquanto que a administração de hidroxietilamido 130/0,4 associado à NaCl 7,5% promove reestabelecimento mais tardio (após 30 minutos), porém mais duradouro (até 120 minutos) da PAM, PAS e PAD em gatas com hipovolemia induzida. A administração de HES 130/0,4 associada à NaCl 7,5% promove aumento acentuado da PVC por até 60 minutos após a administração.

Avaliação laboratorial do uso de solução salina hipertônica e isotônica e de furosemida no tratamento da intoxicação por amônia em bovinos

Para testar a eficiência de vários tratamentos de intoxicação por amônia em bovinos, foram utilizados 25 garrotes que receberam cloreto de amônio por infusão intravenosa (iv) até o surgimento de quadro convulsivo. em seguida, os animais foram alocados em um dos cinco grupos experimentais e tratados da seguinte forma: 1) controle: infusão (iv) de 300mL de solução salina isotônica (SSI), no decorrer de 4h; 2) infusão (iv) de 30mL kg-1 PV de SSI no decorrer de 4h e administração de 4L de água intraruminal por meio de sonda esofágica (ASE); 3) mesmo tratamento do grupo 2 e dose única (iv) de furosemida (2mg kg-1 PV) (F); 4) injeção (iv) de 5mL kg-1 PV de solução salina hipertônica (SSH) 7,2% nos primeiros 30min, seguida de 20mL kg-1 PV de SSI e 4L de ASE; 5) mesmo tratamento do grupo 4 e dose única de F. No decorrer de 4h após a convulsão, foram determinados os teores plasmáticos de amônia e glicose, ureia, creatinina, potássio e sódio séricos, volume e gravidade específica da urina, e excreção urinária de amônio e ureia. No momento da convulsão, os teores de amônia plasmáticos foram muito altos e idênticos em todos os tratamentos, mas no 120°min, nos grupos tratados com associação de SSH+SSI+ASE (grupos 4 e 5), houve diminuição desse metabólito. O uso de furosemida (grupos 3 e 5) não aumentou a excreção total de urina. A terapia com associação de SSH+SSI+ASE aumentou ainda o volume urinário e a excreção percentual urinária de ureia e amônia durante o período crítico da 1ª hora de tratamento, mas o uso de SSI+ASE (grupos 2 e 3) teve resultados intermediários. A eficiência do tratamento com SSH+SSI+ASE ou SSI+ASE foi superior ao grupo controle. Embora com efeito menor que o observado com SSH+SSI+ASE, a SSI+ASE promoveu melhora no quadro clínico geral e, ao término do experimento, promoveu também uma adequada desintoxicação da amônia.

Co-feeding of pacu, Piaractus mesopotamicus Holmberg (1887), larvae with Artemia naupili and a microencapsulated diet

Aiming at a precocious substitution of live prey by artificial diet, a 20-day experiment with pacu, Piaractus mesopotamicus, larvae using co-feeding and abrupt weaning strategies was set up. At the end of the experiment, larvae fed Artemia showed the best results (P < 0.05) in weight, total length and biomass, compared with other treatments. Larvae fed exclusively a microencapsulated diet never ingested the diet. Diet ingestion in co-fed and abrupt-weaned larvae was low, but did increase during the experiment; however, Artemia influenced diet ingestion on co-fed larvae. Careful considerations should be given to diet processing and formulation to ensure survival and growth of larvae fed exclusively on prepared diets. © 2005 by The Haworth Press, Inc. All rights reserved.

Crescimento inicial do cafeeiro irrigado com água salina e salinização do solo

The coffee crop is expanding to new areas with not enough studies about its response to saline irrigation water. The initial growth of coffee plant was evaluated, in greenhouse at the Engineering Department of the Federal University of Lavras (UFLA), under different levels of irrigation water salinity. The completely randomized design was used with 6 treatments (S0 = 0.0 dS m -1, S1 = 0.6 dS m -1, S2 = 1.2 dS m -1, S3 = 1.8 dS m -1, S4 = 2.4 dS m -1 and S5 = 3.0 dS m -1) and 4 replications. The irrigation was accomplished according to soil water retention curve and resistance block reading, restoring the soil water content to its field capacity. It was verified that water salinity affected the plants characteristics significantly. The water salinity above 1.2 dS m -1 caused damage to plant development resulting, in some cases, in death of plants. The leaf area of plant was the variable most affected by salinity of irrigation water. By the end of the experiment, the soil was classified as saline-sodic.

Artemia sp. na alimentação de larvas de jundiá (Rhamdia quelen)

A considerable problem in the catfish larvae rearing is related to the first feeding. This study aimed to determine the larvae growth and the survival of silver catfish submitted to feeding with Artemia sp. in different time periods. The study was conducted in the Aquaculture Laboratory of the Universidade Estadual do Oeste do Paraná, with four treatments and five replications, constituted by feeding with Artemia sp. in 5, 10, 15 and 20 days, and after each period of time, by its replacement by ration. We used 200 larvae distributed into 20 aquariums, soon after the eggs hatched. The larvae were fed 4 times a day until apparent satiation for 30 days. The Artemia sp. feeding for 15 days provided the best results on weight gain, final length and survival.

Uma contribuição para o estudo do efeito causado pela névoa salina em isoladores cerâmicos de chaves fusíveis

Pós-graduação em Ciência e Tecnologia de Materiais - FC