Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante

Since the first description of sulfated polysaccharides from seaweeds, the biological activities of these compounds have been evaluated under different aspects and experimental procedures. Among the broad biological activities presented by seaweed polysaccharides, anticoagulant action appears as a promising function. In this present study we have obtained sulfated polysaccharides from the green seaweed Codium isthmocladium by proteolytic digestion, followed by separation into five fractions (0.3, 0.5, 0.7, 0.9 and 1.2) by sequential acetone precipitation. The chemical analyses have demonstrated that all fractions are composed mainly by sulfated polysaccharides. The anticoagulant activity of these fractions was determined by activated partial thromboplastin time (aPTT) and prothrombin time test (PT) using citrate normal human plasma. None fraction has shown anticoagulant activity by PT test. Furthermore, all of them have shown anticoagulant activity by aPTT test. These results indicated that the molecular targets of these sulfated polysaccharides are mainly in the intrinsic via of the coagulation cascade. Agarose gel electrophoresis in 1,3-diaminopropane acetate buffer, pH 9.0, stained with 0.1% toluidine blue showed the presence of two or three bands in several fractions while the fraction 0.9 showed a single spot. By anion exchange chromatography, the acid polysaccharides from the 0.9 acetone fraction were separated into two new fractions eluted respectively with 2.0 and 3.0 M NaCl. These compounds showed a molecular weight of 6.4 and 7.4 kDa respectively. Chemical analyses and infrared spectroscopy showed that Gal 1 and Gal 2 are sulfated homogalactans and differ one from the other in degree and localization of sulfate groups. aPPT test demonstrated that fractions 2,0 and 3,0M (Gal1 and Gal 2, respectively) have anticoagulant activity. This is the first time that anticoagulant sulfated homogalatans have been isolated from green algae. To prolong the coagulation time to double the baseline value in the aPTT, the required amount of sulfated galactan 1 (6,3mg) was similar to low molecular heparin Clexane®, whereas only 0,7mg of sulfated galactan 2 was needed to obtain the same effect. Sulfated galactan 2 in high doses (250mg) induces platelet aggregation. These results suggest that these galactans from C. isthmocladum have a potential application as an anticoagulant drug

Dinâmica populacional e alimentar de Stegastes fuscus, (Osteichthyes:pomacentridae) em arrecifes da praia de Búzius, no Rio Grande do Norte

The damselfish, Stegastes fuscus Cuvier, 1830 (Osteichthyes: Perciformes: Pomacentridae), is abundant in the coastal reefs of Búzios Beach, Rio Grande do Norte, Brazil and they play an important role in the reef community ecology. The present study investigated the feeding strategy of this species considering the food habits and morfo-histology of the digestive tract. Influence of the environmental correlates such as temperature, rainfall and luminosity in the tidal rock pools were studied. The fish were captured on a monthly basis from September 2004 to August 2005, during which period 842 individuals of S. fuscus were captured, 125 males, 437 females and 280 individuals without sex identification. The sex ratio observed was 1 M : 3,5 F. The total body length of males varied from 3.6 to 11.3 cm, with a mean of 7.77 cm; that of females varied from 2.9 to 11.4 cm, with a mean of 7.85 cm, and that of sex grouped individuals varied from 2.9 to 11.4 cm, with a mean of 7.83. However, there was no difference between males and females in total body length. This species presented a positive alometric growth and the equations obtained for the relation between body mass and total body length were: Wt =0,0174Lt 3,1123 for males; Wt =0,0137Lt 3,2294 for females and Wt = 0,0148Lt 3.1928 for sex grouped individuals. The relation between total body length and standard length was L t = 1 ,3223Ls + 0,1527 for sex grouped individual s. February to August was associated to a long period of gonadal resting. The fish spawned during January and in September -October. The frequency of fish with empty stomachs occurred during August to December, whereas frequency of fish with food contents i n stomachs occurred during January to July. This species is considered as a preferential herbivore based on the volume of macroalgae in its diet composition. The morfo -histological aspects of S. fuscus confirm herbivory. Among the environmental factors con sidered only rainfall showed a correlation with the feeding habits of this species

Efeito imunomodulador e atividade antimicrobiana de heterofucanas de Sargassum filipendula

Fucans, sulphated polysaccharides that contain L-fucose in its constitution, obtained from species of Phaeophyceae of the Sargassum kind, display several biological activities. Heterofucans from Sargassum filipendula are bioactive molecules that contain strong antiproliferative and antioxidant activity. However, their immunomodulatory and antimicrobial activities have not yet been examined. In this context, the aim of this research was to evaluate the heterofucans as for their immunomodulatory capacity and antimicrobial action against Leishmania infantum, Trichomonas vaginalis, Staphylococcus epidermidis and Klebsiella pneumonia (KPC). The five heterofucans obtained from S. filipendula show activities that are distant as stimulants of the immune system and microbial agent. The SF0.5V, SF0.7V amd SF1.0V heterofucans were capable of acting in the activation of murine and human macrophages. In addition to that, SF0.5V has shown antibiofilm activity of S. epidermides and SF0.7V and 1.0V almost completely inhibited the survival of the protozoan T. vaginalis. Results such as this one, reflect the broad range of action of the sulphated polysaccharides obtained from seaweeds, especially from the species S.filipendula

Estudo comparativo de dois tipos de cultivo: monocultivo (camarões) versus cultivo integrado (algas/camarões)

The integrated culture of seaweed and aquatic animals is an ancient practice in Asian countries. The expansion of this practice to western countries is consequence of the recognition of this system as a sustainable alternative that allows economical diversification and mitigation of environmental impacts generated by effluents of aquaculture. This study evaluated the growth of the seaweed Gracilaria caudata and of the shrimp Litopenaeus vannamei in monoculture (shrimps) and integrated culture (shrimps and algae) systems, and accessed the effect of the seaweed in the water quality. There were two treatments in the experiment: monoculture (shrimps) and integrated culture (shrimps/ algae). The organisms were cultured in 6 aquaria (10L) filled with seawater (35.0±0.0 PSU and 28.1±0.4°C) for 28 days. The nutrients of water (PO43-, NH4+, NO2-, NO3- and DIN), the biomass and the relative growth rate (RGR, % day-1) of seaweed and shrimps were measured weekly. The parameters pH, temperature, salinity and dissolved oxygen were measured daily. The concentration of NH4+ in integrated culture (62.8±25.2µM) was lower (Mann-Whitney p<0.001) than in monoculture (85.6±24.3µM). The mean of PO4- in monoculture (10.4±4.6µM) was markedly higher (Mann-Whitney; p=0.024) than that in integrated culture (8.7±4.1µM). The level of dissolved oxygen in integrated culture (6.0±0.6mg/L) was higher (t-Student; P=0.014) than that in shrimp monoculture (5.8±0.6mg/L). The mean values of the parameters pH, NO2-, NO3- and DIN were 7.5±0.2, 10.1±12.2µM, 24.5±3.2µM and 120.17±30.76µM in monoculture, and 7.5±0.2, 10.5±13.2µM, 27.4±3.5µM and 100.76±49.59µM in integrated culture. There were not differences in these parameters between treatments. The biomass and RGR of seaweed reached 15.0±1.9g and 7.4±2.8% day-1 at the end of the experiment. The performance of shrimp was favorable in monoculture (1.5±0.8g; 5.7±1.6% dia-1) and in integrated culture (1.5±0.7g; 5.2±1.2% dia-1), and the rate of survival was 100% in both treatments. The tolerance and favorable performance of Gracilaria caudata suggest that this seaweed might be integrated into shrimp (Litopenaeus vannamei) culture systems

Influência da profundidade e dos parâmetros ambientais sobre o desenvolvimento de Gracilaria birdiae (Rhodophyta, Gracilariales) em viveiro de camarão

This study evaluates the influence of depth and environmental parameters on the development of Gracilaria birdiae Plastino & Oliveira (Gracilariaceae Rhodophyta) in an organic shrimp pound (Litopenaeus vannamei) under euthrophical conditions. PVC structures (module) witch four ropes laden with 150 g of macroalgae each, were kept during 35 days at three different depths (surface, 10 and 20 cm depth). Wet biomass weighing and environmental parameters (temperature, salinity, turbidity, pH, transparence, precipitation, evaporation, insolation, accumulated solar radiation, nitrite, nitrate, ammonium and orthophosphate) were measured weekly. At all three proposed depths, the macroalgae displayed a higher biomass at the end of experiment than at the initial inoculations. The module kept at a 10 cm depth presented the greatest average biomass (186,3), followed by that kept at 20 cm (180,4 g) and the surface module (169,9 g). Biomass variations showed algae to suffer the direct effects of depths. Biomass loss was associated with the factors that influence light penetration, such as sediment deposits above the thallus, rate of evaporation and precipitation. The smallest loses occurred in the algae kept on surface (0,16%), followed by the algae kept at 20 cm (0,20%) and 10 cm (0,22%). The specific growth rate (SGR) of G. birdiae showed no significant difference between the three depths nor the sample periods. Nevertheless, the modules kept at 10 and 20 cm depths presented similar growth evolution, both growing 0,38%·per day-1, while the module kept on surface had an average SGR of 0,36%·day-1. The models related to growth rate demonstrated temperature, salinity, pH, orthophosphate, ammonium, precipitation and turbidity as the principal environmental parameters influencing the development of G. birdiae

Efeito dos nutrientes (nitrogênio e fósforo) sobre o desenvolvimento da Gracilaria birdiae: crescimento e possível uso como biofiltro

Marine shrimp farming has grown exponentially during the last years in Brazil. In spite of the promising economical situation, this activity is facing an increasing criticism due to its environmental impact. Thus, the necessity of alternatives to mitigate environmental degradation caused by this activity. An alternative that is being studied is the policulture that is the integrated culture of two or more organisms, normally one of them a filtering organism. Among filtering organisms, macroalgae are very practicable because they are efficient in the removal of the exceeding nutrients of the water and do not leave residues in the water. Besides, the integrated culture with macroalgae allows the economical exploration of the seaweed (for the manufacture of jelly and jam, for the dairy industry, pharmaceuticals, etc.) along with possibility of a sustainable aquaculture. In the present experiment, the development of the seaweed Gracilaria birdiae, the influence and tolerance of this species to the environmental parameters, and its absorption efficiency in relation with the three kinds of macronutrients (NH4+, NO3- and PO4-3) found in the effluents of marine shrimp farming was studied. The experiment was divided in two parts: a laboratorial part and one part carried under natural conditions. The water used in the laboratory trial was collected in the shrimp ponds of Tecnarão farm and distributed in aquaria containing 20 g of G. birdiae. In the field trial, 0.5 kg of G. birdiae was inserted in PVC cages cultivated in the farm. The results of the study showed a modest growth of G. birdiae, probably due to its low tolerance to highly eutrophicated environments. However, the removal of nutrients was very expressive. Ammonia was reduced in approximately 34 %. Ortho-phosphate showed a reduction of 93.5 %. The capacity of biofiltration of the NO3- by the macro algae was of 100 %, showing that G. birdiae is a seaweed-filtered with a high level of removal for this nutrient under laboratorial conditions. In spite of the low growth of the macro algae in the experiment, the results in relation to the removal of nutrients of the water was encouraging, suggesting that this species can be an efficient biofilter and thus, a strong candidate to be used in a sustainable aquaculture

Utilização da macroalga Ulva lactuca linnaeus na redução de nutrientes (NH4+,NO3- e PO4-2) provenientes da carcinicultura

Dentre as macroalgas capazes de absorver altas concentrações de N e P dissolvidos na água, destaca-se a Chlorophyta Ulva lactuca, bastante adaptável e resistente às adversidades ambientais, como grandes variações de temperatura, salinidade, matéria orgânica e metais pesados. Trata-se também de uma espécie bastante comum nas áreas intertidais do litoral norte-riograndense. Devido a suas características ecológicas, fisiológicas e nutricionais, foi avaliado nesse estudo, o seu potencial como biofiltro na redução de NH4+, NO3- e PO4-2, tanto em condições controladas como também em um viveiro de camarão. No experimento laboratorial, foram utilizados quatro aquários de vidro de 30 x 20 x 20cm com 10L de água, sendo três aquários experimentais contendo 20g de U. lactuca e um controle. O acréscimo de biomassa foi de 2,92g (22,92 ± 6,29g; p < 0,05) em relação ao inóculo inicial de 20g, sob temperatura (28,50 ± 0,58ºC), salinidade (35,00 ± 0,00 ), pH (8,26 ± 0,02) e luz constante (250 μmol.m2s-1). O crescimento positivo (1,78 ± 4,38%dia-1; p < 0,05), juntamente com a alta eficiência de absorção de amônio (83%; p < 0,001), nitrato (83%; p < 0,001) e ortofosfato (53%; p < 0,001), demonstrou que, nessas condições, a Ulva lactuca absorveu os nutrientes e aumentou sua biomassa. Já no experimento de campo, realizado na fazenda TECNARÃO, situada no município de Arez/RN (06° 11 40 Latitude Sul, e 35º 09 37 Longitude Oeste), foram utilizadas três gaiolas de PVC, posicionadas a 12cm da superfície da água, cada uma com dimensões de aproximadamente 59 x 59 x 15cm, onde foram colocadas 200g de U. lactuca. O ganho de biomassa de 3g (203,00 ± 41,02g; p < 0,001) foi muito semelhante às condições controladas, demonstrando a adaptabilidade da espécie em condições ambientais variáveis, onde, apesar da temperatura pouco variável (27,45 ± 0,64ºC), houve progressiva diminuição de salinidade (25 - 15 ), devido ao período de fortes chuvas (34,70 ± 23,78mm). Somado a isso, foram observados vários fatores biológicos interferindo no viveiro, como a presença de epífitas, organismos endofíticos, fouling e a herbivoria por parte dos próprios camarões. Houve aumento nas concentrações de NH4+ (4,36 ± 1,69 μmol.L-1), NO3- (0,17 ± 0,25μmol.L-1) e PO4-2 (0,41 ± 0,13μmol.L-1), coincidindo com o crescimento da espécie até a terceira semana. Todos os parâmetros ambientais analisados, assim como a biomassa e a Taxa de Crescimento Relativo (TCR), obtidos no campo, apresentaram variações altamente significativas (p < 0,001). As correlações observadas entre biomasa e NH4+ (r = 0,82; p < 0,001) e entre biomassa e PO4-2 (r = 0,87; p < 0,001), indicam que esta espécie é capaz de ter um crescimento satisfatório nas condições eutróficas de um viveiro de camarão, sendo possível seu uso como biofiltro.

Biodiversidade e abundância da ictiofauna associada ao cultivo orgânico de Litopenaeus vannamei (Boone, 1931)

The expansion of shrimp farming has caused a series of environmental impacts, often as a result of lack of planning and adequate management. Organic aquaculture has emerged as an alternative to conventional shrimp farming, and differently, aims at the economical, ecological and farming potential of other organisms, such as fishes, oysters and seaweeds. The present study aimed at evaluating the biological diversity and abundance of the ichthyofauna associated to Litopenaeus vannamei organic culture at PRIMAR farm (Rio Grande do Norte, Brazil). The sampling period consisted of four culture cycles (1, 2, 3, and 4) accomplished in four grow-out ponds (V1, V7, V2, and V4) in 2005. The first two culture cycles were carried out during the rainy season, and the last two cycles, during the dry season. Environmental (temperature, salinity, transparency, and rainfall), biodiversity (diversity and uniformity and abundance indexes) of the ichthyofauna, and shrimp productivity data were collected throughout these four culture cycles. The results obtained for the environmental variables showed that both salinity and transparency oscillated in a significant way (p < 0.001) between culture cycles. In relation to the abundance of the ichthyofauna, fifty four species associated to the organic culture of Litopenaeus vannamei were collected and identified at PRIMAR. For the biodiversity criterion, larger species richness (S = 46) and uniformity (E = 0.59 ± 0.16) were observed during cycles 3 and 4 than in cycles 1 and 2 (S = 41 and E = 0.15 ± 0.12). A positive and significant correlation was obtained for the transparency with diversity and uniformity of fish species between cycles 1 and 2 (wet season) and cycles 3 and 4 (dry season) (p < 0.017 - Shannon Index; p < 0.008 - Pielou Index, respectively). A correlation could not be established between shrimp productivity and fish biomass. However, fish biomass decreased from cycles 1 and 2 to cycles 3 and 4, concomitant with an increase in shrimp productivity, most probably due to a higher (from 58.0% to 71.0%) shrimp survival. Regarding the culture potential of the ichthyofauna, three species (Mugil curema, Mugil liza and Chaetodipterus faber) were identified as potential farming alternatives, either singly or in consortium with Litopenaeus vannamei. A fourth species (Centropomus undecimalis) was indicated as an alternative for single culture or in consortium with other fish species. In conclusion, the large diversity of estuarine fishes associated to Litopenaeus vannamei farming observed at PRIMAR clearly indicated the ecological feasibility for organic aquaculture in northeastern Brazil

Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante

Since the first description of sulfated polysaccharides from seaweeds, the biological activities of these compounds have been evaluated under different aspects and experimental procedures. Among the broad biological activities presented by seaweed polysaccharides, anticoagulant action appears as a promising function. In this present study we have obtained sulfated polysaccharides from the green seaweed Codium isthmocladium by proteolytic digestion, followed by separation into five fractions (0.3, 0.5, 0.7, 0.9 and 1.2) by sequential acetone precipitation. The chemical analyses have demonstrated that all fractions are composed mainly by sulfated polysaccharides. The anticoagulant activity of these fractions was determined by activated partial thromboplastin time (aPTT) and prothrombin time test (PT) using citrate normal human plasma. None fraction has shown anticoagulant activity by PT test. Furthermore, all of them have shown anticoagulant activity by aPTT test. These results indicated that the molecular targets of these sulfated polysaccharides are mainly in the intrinsic via of the coagulation cascade. Agarose gel electrophoresis in 1,3-diaminopropane acetate buffer, pH 9.0, stained with 0.1% toluidine blue showed the presence of two or three bands in several fractions while the fraction 0.9 showed a single spot. By anion exchange chromatography, the acid polysaccharides from the 0.9 acetone fraction were separated into two new fractions eluted respectively with 2.0 and 3.0 M NaCl. These compounds showed a molecular weight of 6.4 and 7.4 kDa respectively. Chemical analyses and infrared spectroscopy showed that Gal 1 and Gal 2 are sulfated homogalactans and differ one from the other in degree and localization of sulfate groups. aPPT test demonstrated that fractions 2,0 and 3,0M (Gal1 and Gal 2, respectively) have anticoagulant activity. This is the first time that anticoagulant sulfated homogalatans have been isolated from green algae. To prolong the coagulation time to double the baseline value in the aPTT, the required amount of sulfated galactan 1 (6,3mg) was similar to low molecular heparin Clexane®, whereas only 0,7mg of sulfated galactan 2 was needed to obtain the same effect. Sulfated galactan 2 in high doses (250mg) induces platelet aggregation. These results suggest that these galactans from C. isthmocladum have a potential application as an anticoagulant drug