Purificação e caracterização parcial de uma B-D glicosidade de Artemia franciscana com ação sobre celobiose e lactose

-D-glucosidase (EC 3.2.1.21) is one of the most interesting glycosidases, especially for hydrolysis cellobiose releasing glucose, is last step degradation of cellulose. This function makes the -D-glucosidase is of great interest as a versatile industrial biocatalyst, being critical to various bio-treatment / biorefinery processes, such as bioethanol production. Hen in the report, a -D-glucosidase was extracts from protein extracted of the invertebrate marine Artemia franciscana was purified and characterized with a combination of precipitation with ammonium sulfate (0 - 30%, 30 to 50%, 50 to 80%), the fraction saturated in the range of 30 to 50% (called F-II) was applied in a molecular exclusion chromatography, in Sephacryl S-200, the fractions corresponding to the first peak of activity of -D-glucosidase were gathered and applied in a chromatography of ion exchange in Mono Q; the third peak this protein obtained chromatography, which coincides with the peak of activity of -D-glucosidase was held and applied in a gel filtration chromatography Superose 12 where the first peak protein, which has activity of -D-glucosidase was rechromatography on Superose 12. This enzyme is probably multimerica, consisting of three subunit molecular mass of 52.7 kDa (determined by SDS-PAGE) with native molecular mass of 157 kDa (determined by gel filtration chromatography on Superose 12 under the system FPLC). The enzyme was purified 44.09 times with a recovery of 1.01%. Using up p-nitrophenyl-β-D-glucopiranoside as substrate obtained a Km apparent of 0.229 mM and a Vmax of 1.109 mM.60min-1.mL-1mM. The optimum pH and optimum temperature of catalysis of the synthetic substrate were 5.0 and 45 °C, respectively. The activity of the -D-glucosidase was strongly, inhibited by silver nitrate and N- etylmaleimide, this inhibition indicates the involvement of radical sulfidrila the hydrolysis of synthetic substrate. The -D-glucosidase of Artemia franciscana presented degradativa action on celobiose, lactose and on the synthetic substrate -nitrophenyl-β-D-glucopiranoside indicating potential use of this enzyme in the industry mainly for the production of bioethanol (production of alcohol from the participating cellulose), and production hydrolysate milk (devoid of milk lactose)

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.

Crescimento e estruturas do sistema digestório de larvas de pacu alimentadas com dieta microencapsulada produzida experimentalmente

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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)

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.

Estudo fitoquímico preliminar e bioensaio toxicológico frente a larvas de Artemia salina Leach. de extrato obtido de frutos de Solanum lycocarpum A. St.-Hill (Solanaceae)

The phytochemical profile of ethanolic extract of Solanum lycocarpum fruits was analyzed and preliminary toxicity tests were performed against brine shrimp larvae. The extract was subjected to preliminary phytochemical analysis to identify the main classes of secondary metabolites and tested against the larvae of A. salina to obtain the median lethal concentrations (LC50%). The phytochemical tests showed the presence of phenols, tannins, saponins, alkaloids and free steroids. The extract was fractionated with various solvents for toxicity testing against the larvae and the hydroalcoholic fraction showed considerable cytotoxicity (CL50% = 285.546 g/mL).

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.

Efeito de dietas e altura da coluna d’água na sobrevivência de larvas de Betta splendens e o aporte de nitrogênio e fósforo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo de biorremediação dos azo corantes têxteis Acid Blue 161 e Procion Red MX-5B por fungos filamentosos em solução simples e solução binária associado a testes de toxicidade com Lactuca sativa e Artemia salina

Pós-graduação em Ciências Biológicas (Microbiologia Aplicada) - IBRC

Dietas microencapsuladas: produção e avaliação para alimentação de larvas altriciais de peixes de água doce

Pós-graduação em Zootecnia - FCAV

Feeding adult of Artemia salina (Crustacea-Branchiopoda) on the dinoflagellate Gyrodinium corsicum (Gymnodiniales) and the Chryptophyta Rhodomonas baltica

Experimentos foram desenvolvidos para estudar as taxas de alimentação e de sobrevivência de Artemia salina alimentada com cepas não tóxicas do dinoflagelado Gyrodinium corsicum e da Chryptophyta Rhodomonas baltica. As taxas de filtração sobre R. baltica e G. corsicum variaram entre 3,35 e 7,14 ml.artemia-1.h-1 e 2,97 e 15,86 ml.artemia-1.h-, respectivamente. As taxas de ingestão observadas para A. salina não indicaram disfunção digestiva ou prejuízo fisiológico nos organismos alimentados com G. corsicum, sendo a resposta funcional destes organismos similar a observada em copépodos alimentados com diferentes concentrações de alimento. As taxas de mortalidade de A. salina oscilaram entre 2,5 e 100% quando alimentada com R. baltica e G. corsicum, respectivamente. As maiores taxas de mortalidade observadas para os organismos alimentados com G. corsicum indicam que este dinoflagelado apresenta algum efeito nocivo sobre A. salina, embora não tenha sido possível confirmar se sua origem está relacionada com a produção de toxinas ou com a inadequação nutritiva deste dinoflagelado para alimentação de organismos desta espécie.

Probiótico na larvicultura de matrinxã, Brycon amazonicus

The aim of this study was to evaluate the growth performance of Brycon amazonicus larvae fed with probiotic consisting of Bacillus subtilis, bioencapsulated in Artemia salina at different doses. Six treatments were used: T0 = control (without probiotic), T2.5 = 2.5 g, T5.0 = 5.0 g, T7.5 = 7.5 g, T10.0 = 10.0 g and T12;5 = 12.5 g per liter of probiotic when fed with A. saline (4h before feeding the larvae); and per kilogram of feed (commercial powder with 40% crude protein) in the next phase. The results showed heterogeneity with regard to growth, indicating that larvae fed with probiotic had no influence on cannibalism.