Extração, caracterização estrutural parcial e atividades farmacológicas do olginato obtido da alga marrom Lobophora variegata (Lamouroux) Oliveira Filho, 1977

The alginic acid or alginates are acidic polysaccharides found in brown seaweed widely used in food, cosmetic, medical and pharmaceutical industry. This paper proposes the extraction, chemical characterization and verification of the pharmacological activities of brown seaweed variegata Lobophora . The alginate was extracted from the seaweed Lobophora variegata and part was sulphated for comparative purposes. The native extract showed 42% total sugar, 65% uronic acid, 0,36 % protein and 0% of sulfate, while the sulfate showed 39% , 60%, 0.36% and 27,92 % respectively. The presence of a sulfate group may be observed by the metachromasia with toluidine blue in electrophoresis system and characteristic vibration 1262,34 cm-1 in infrared spectroscopy connections assigned to S = O. We observed the formation of films and beads of native alginate, where more concentrated solution 6% resulted in a thicker and more consistent film. Native alginate showed proliferative activity at concentrations (25 and 50 mcg), (50 mg) and (100 mg) in 3T3 cell line in 24h, 48h and 72h, respectively , as the sulfated (100 mg) in 24 . Also showed antiproliferative or cytotoxic activity in HeLa cells of strain, (25 and 100 mg), (25 and 100 mg) and (25, 50 and 100 mg), to native, now for the sulfate concentrations (100 mg) in 24 (25, 50 and 100 mg) in 48 hours, and (50 and 100 mg ) 72h. For their antioxidant activity, the sulfated alginates have better total antioxidant activity reaching 29 % of the native activity while 7.5 % of activity . For the hydroxyl radical AS showed high inhibition ( between 77-83 % ) in concentrations, but the AN surpassed these numbers in the order of 78-92 % inhibition. The reducing power of AN and AS ranged between 39-82 % . In the method of ferric chelation NA reached 100 % chelating while the AS remained at a plateau oscillating 6.5%. However, in this study , we found alginates with promising pharmacological activities, to use in various industries as an antioxidant / anti-tumor compound

Dual Quorum Quenching Capsules: Disrupting two bacterial communication pathways that lead to virulence

Healthcare Associated Infections (HAIs) in the United States, are estimated to cost nearly $10 billion annually. And, while device-related infections have decreased, the 60% attributed to pneumonia, gastrointestinal pathogens and surgical site infections (SSIs) remain prevalent. Furthermore, these are often complicated by antibacterial resistance that ultimately cause 2 million illnesses and 23,000 deaths in the US annually. Antibacterial resistance is an issue increasing in severity as existing antibiotics are losing effectiveness, and fewer new antibiotics are being developed. As a result, new methods of combating bacterial virulence are required. Modulating communications of bacteria can alter phenotype, such as biofilm formation and toxin production. Disrupting these communications provides a means of controlling virulence without directly interacting with the bacteria of interest, a strategy contrary to traditional antibiotics. Inter- and intra-species bacterial communication is commonly called quorum sensing because the communication molecules have been linked to phenotypic changes based on bacterial population dynamics. By disrupting the communication, a method called ‘quorum quenching’, bacterial phenotype can be altered. Virulence of bacteria is both population and species dependent; each species will secrete different toxic molecules, and total population will affect bacterial phenotype9. Here, the kinase LsrK and lactonase SsoPox were combined to simultaneously disrupt two different communication pathways with direct ties to virulence leading to SSIs, gastrointestinal infection and pneumonia. To deliver these enzymes for site-specific virulence prevention, two naturally occurring polymers were used, chitosan and alginate. Chitosan, from crustacean shells, and alginate, from seaweed, are frequently studied due to their biocompatibility, availability, self-assembly and biodegrading properties and have already been verified in vivo for wound-dressing. In this work, a novel functionalized capsule of quorum quenching enzymes and biocompatible polymers was constructed and demonstrated to have dual-quenching capability. This combination of immobilized enzymes has the potential for preventing biofilm formation and reducing bacterial toxicity in a wide variety of medical and non-medical applications.

Sulphated polysaccharides from Ulva clathrata and Cladosiphon okamuranus seaweeds both inhibit viral attachment/entry and cell-cell fusion, in NDV infection

Sulphated polysaccharides (SP) extracted from seaweeds have antiviral properties and are much less cytotoxic than conventional drugs, but little is known about their mode of action. Combination antiviral chemotherapy may offer advantages over single agent therapy, increasing efficiency, potency and delaying the emergence of resistant virus. The paramyxoviridae family includes pathogens causing morbidity and mortality worldwide in humans and animals, such as the Newcastle Disease Virus (NDV) in poultry. This study aims at determining the antiviral activity and mechanism of action in vitro of an ulvan (SP from the green seaweed Ulva clathrata), and of its mixture with a fucoidan (SP from Cladosiphon okamuranus), against La Sota NDV strain. The ulvan antiviral activity was tested using syncytia formation, exhibiting an IC50 of 0.1 μg/mL; ulvan had a better anti cell-cell spread effect than that previously shown for fucoidan, and inhibited cell-cell fusion via a direct effect on the F0 protein, but did not show any virucidal effect. The mixture of ulvan and fucoidan showed a greater anti-spread effect than SPs alone, but ulvan antagonizes the effect of fucoidan on the viral attachment/entry. Both SPs may be promising antivirals against paramyxovirus infection but their mixture has no clear synergistic advantage

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

Variação no conteúdo protéico e pigmentar em variantes cromáticas de Gracilaria domingensis nas populações naturais de Rio do Fogo-RN, BRASIL

The seaweed Gracilaria domingensis is a common species in the coast of Rio Grande do Norte. This species lives in the intertidal zone, where colour strains (red, green and brown) co-occur during the whole year. Seaweeds that live in this region are exposed to daily changes and to the rhythm of the tide. During the low tide they are exposed to dissection, hiper-or hipo-osmotic shock, high temperatures and high irradiance. The aim of this study was to analyze whether the pigment and protein content of the colour strains of G. domingensis is affected by some environmental parameters in a temporal scale. The seaweeds were collected during 10 months in the seashore of Rio do Fogo (RN). The total soluble proteins and the phycobiliprotein were extracted in phosphate buffer and the carotenoids were analyzed by a standardized method through HPLC-UV. The pigments analysis showed that phycoerithrin is the most abundant pigment in the three strains. This pigment was strongly correlated with nitrogen and the photosynthetically active radiation. Chlorophyll presented higher concentrations than carotenoids during the whole, but the ratio carotenoid/chlorophyll-a was modified by incident radiation. The most abundant carotenoid was ß-carotene and zeaxanthin, which had higher concentrations in the higher radiation months. The concentration increase of zeaxanthin in this period indicated a photoprotective response of the seaweed. The three strains presented a pigment profile that indicates different radiation tolerance profile. Our results pointed that the green strain is better adapted to high irradiance levels than the red and brown strains

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.

Estudo do crescimento, eficiência de biofiltração e cinética de absorção de nutrientes (N-NH, N-NO e P-PO4³) da macroalga Gracilaria cervicornis (Turner) J. Agardh

The objective of this study was to examine the growth of Gracilaria cervicornis cultured in a shrimp (Litopenaeus vannamei) pond and to determine the absorption efficiency and the kinetics parameters (Vmax, Ks e Vmax:Ks) of this macroalgae for the nutrients N-NO3-, N-NH4+ and P-PO4-3, aiming at its use as bioremediatory of eutrophicated environments. For this study, two experiments (field and laboratory) were developed. In the field study, the seaweed was examined in relation to the growth and the biomass. In the laboratory experiment, the absorption efficiency of G. cervicornis was measured through the monitoring of the concentration of the three nutrients (N-NO3-, N-NH4+ e P-PO4-3) during 5 hours and the kinetic parameters were determined through the formula of Michaelis-Menten. The results obtained in this study demonstrated that G. cervicornis benefited from the available nutrients in the pond, increasing 52.4% of its biomass value after 30 days of culture. It was evidenced that the variability of the biomass could be explained through the salinity, availability of light (transparency and solid particle in suspension) and concentration of N-NO3- in the environment. In the laboratory experiment, the highest absorption efficiency was found in the treatments with low concentration (5 µmol.L-1), being evidenced a reduction of up to 85,3%, 97,5% and 81,2% of N-NH4+, N-NO3- and P-PO43-, respectively. Regarding the kinetic parameters, G. cervicornis presented better ability in absorbing N-NH4+ in high concentrations (Vmax = 158,5 µmol g-1 dry wt h-1) and P-PO43- in low concentrations (Ks = 5 µmol.L-1 e Vmax:Ks = 10,3). The results of this study show that G. cervicornis could be cultivated in shrimp ponds, presents a good capacity of absorption for the tested nutrients and is a promising candidate for biorremediation in shrimp pond effluent

Potencialidades de cultivo de macroalgas no litoral do Rio Grande do Norte (NE, Brasil): uma perspectiva de integração de dados ambientais em sistemas de informação geográfica (SIG)

Since centuries ago, the Asians use seaweed as an important source of feeding and are their greatest world-wide consumers. The migration of these peoples for other countries, made the demand for seaweed to increase. This increasing demand prompted an industry with annual values of around US$ 6 billion. The algal biomass used for the industry is collected in natural reservoirs or cultivated. The market necessity for products of the seaweed base promotes an unsustainable exploration of the natural banks, compromising its associated biological balance. In this context, seaweed culture appears as a viable alternative to prevent the depletion of these natural supplies. Geographic Information Systems (GIS) provide space and produce information that can facilitate the evaluation of important physical and socio-economic characteristics for the planning of seaweed culture. This objective of this study is to identify potential coastal areas for seaweed culture in the state of Rio Grande do Norte, from the integration of social-environmental data in the SIG. In order to achieve this objective, a geo-referred database composed of geographical maps, nautical maps and orbital digital images was assembled; and a bank of attributes including physical and oceanographical variables (winds, chains, bathymetry, operational distance from the culture) and social and environmental factors (main income, experience with seaweed harvesting, demographic density, proximity of the sheltered coast and distance of the banks) was produced. In the modeling of the data, the integration of the space database with the bank of attributes for the attainment of the map of potentiality of seaweed culture was carried out. Of a total of 2,011 ha analyzed by the GIS for the culture of seaweed, around 34% or 682 ha were indicated as high potential, 55% or 1,101 ha as medium potential, and 11% or 228 ha as low potential. The good indices of potentiality obtained in the localities studied demonstrate that there are adequate conditions for the installation of seaweed culture in the state of Rio Grande do Norte

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

Variation in the impact of non-native seaweeds along gradients of habitat degradation: a meta-analysis and an experimental test

Biological invasions are acknowledged among the main drivers of global changes in biodiversity. Despite compelling evidence of species interactions being strongly regulated by environmental conditions, there is a dearth of studies investi-gating how the effects of non-native species vary among areas exposed to different anthropogenic pressures. Focusing on marine macroalgae, we performed a meta-analysis to test whether and how the direction and magnitude of their effects on resident communities and species varies in relation to cumulative anthropogenic impact levels. The relationship between human impact levels and non-native species impact intensity emerged only for a reduced subset of the response variables examined. Yet, there was a trend for the effects of non-native species on community biomass and abundance and on species abundance to become less negative at heavily impacted sites. By contrast, the magnitude of negative effects of seaweed on community evenness tended to increase with human impact levels. The hypothesis of decreasing severity of invader’ impacts along a gradient of habitat degradation was also tested experimentally at a regional scale by comparing the effects of the removal of non-native alga,
Caulerpa cylindracea, on resident assemblages among rocky reefs exposed to different anthropogenic pressures. Assemblages at urban and pristine site did not differ when invaded, but did so when C. cylindracea was removed. Our results suggest that, despite the generally weak relationship between human impacts levels and non-native species impacts, more negative impacts can be expected in less stressful environments (i.e. less degraded or pristine sites), where competitive interactions are presumably the driving force structuring resident communities. Implementing strategies for controlling the establishment of non-native seaweeds should be, thus, considered a priority for preserving biodiversity in relatively pristine areas. On the other hand, control of invaders at degraded sites could be warranted to lessen their role as propagule sources

Macroalgae and commercial macroalgal polysaccharides as potential functional ingredients in muscle foods

Ulva rigida (UR) and Palmaria palmata (PP) were included in farmed Atlantic salmon diets at levels of 0-15% for 19 and 16 weeks, respectively. Quality and shelf-life parameters of salmon fillets stored in modified atmosphere packs (MAP) (60% N2 : 40% CO2) at 4ºC were compared to controls fed astaxanthin. Salmon fillets were enhanced with a yellow/orange colour. Proximate composition, pH and lipid oxidation were unaffected by dietary UR and PP. Salmon fed 5% UR and 5-15% PP did not influence sensory descriptors (texture, odour, oxidation flavour and overall acceptability) of cooked salmon fillets. Pig diets were supplemented with commercial wet- and spray-dried macroalgal (Laminaria digitata) polysaccharide extracts containing laminarin (L, 500 mg/kg feed) and fucoidan (F, 420 mg/kg feed) (L/F-WS, L/F-SD) for 3 weeks and quality and shelf-life parameters of fresh pork steaks (longissimus thoracis et lumborum) stored in MAP (80% O2 : 20% CO2) were examined. Level (450 or 900 mg L and F/kg feed) and duration (3 or 6 weeks) of dietary L/F-WS and mechanisms of antioxidant activities in pork were investigated. L/F-WS reduced (p < 0.05) lipid oxidation and lowered levels of saturated fatty acids in fresh pork after 3 weeks feeding. L/F-SD was added directly to mince pork (0.01 - 0.5%) and quality and shelf-life parameters of fresh pork patties stored in MAP (80% O2 : 20% CO2) were assessed. Direct addition of the L/F-SD increased levels of lipid oxidation and decreased surface redness (a* values) of fresh pork patties. Lipid oxidation was reduced in cooked patties due to the formation of Maillard reaction products. Cooked pork patties containing L/F-SD were subjected to an in vitro digestion and a cellular transwell model to confirm bioaccessibility and uptake of antioxidant compounds. In mechanistic studies, fucoidan demonstrated antiand pro-oxidant activities on muscle lipids and oxymyoglobin, respectively.

Biomethane production from macroalgae

Irish brown seaweeds have been identified as a potential bio-resource with potentially high specific methane yields. Anaerobic digestion is deemed the most feasible technology due to its commercial viability for handling such wet feedstock. However, the biomethane potential of seaweed is highly dependent on its chemical composition which can vary by species type, cultivation method, and time of harvest. This study aims to investigate and optimize the process for the production of biomethane from Irish brown seaweeds focusing on the key technology bottlenecks including for seaweed characterization, biomethane potential assessment, optimization of long-term anaerobic digestion and suitable pre-treatment technologies to enhance potential gas yields. Laminaria digitata and Ascophyllum nodosum were tested for seasonal variation. From the characterization and batch digestion of L. digitata, August was found to be the optimal month for harvest due to high organic matter content, low level of ash and ultimately highest biomethane yield. The specific methane yield of 53 m3 CH4 t-1 wwt in August was 4.5 times higher than the yield in December (12 m3 CH4 t-1 wwt), with ash content the key factor in seasonal variation. For A. nodosum, the optimal harvest month was October with polyphenol content found to be a more influential factor than ash. The gross energy yields from both species were evaluated in the range of 116-200 GJ ha-1 yr-1. Continuous digestion trials were subsequently designed for S. latissima and L. digitata to optimize the key digestion parameters. Results from mono-digestion and co-digestion with dairy slurry revealed that both seaweeds could be digested at maximum biomethane efficiency to a loading rate of 4 kg VS m-3 d-1. Accumulation of salt in the digesters was a concern for long term digestion and it was reasoned that suitable pretreatment may be required prior to digestion. Various pre-treatments were subsequently tested on L. digitata to enhance the gas yield. It was found that maceration after hot water washing yielded 25% more specific methane and up to 54% salt removal as compared to untreated L. digitata. The experiments undertaken aim to assist in providing a basic guideline for feasible design and operation of seaweed digesters in Ireland.

EUNIS habitat's thresholds for the Western coast of the Iberian Peninsula — A Portuguese case study

The European Nature Information System (EUNIS) has been implemented for the establishment of a marine European habitats inventory. Its hierarchical classification is defined and relies on environmental variables which primarily constrain biological communities (e.g. substrate types, sea energy level, depth and light penetration). The EUNIS habitat classification scheme relies on thresholds (e.g. fraction of light and energy) which are based on expert judgment or on the empirical analysis of the above environmental data. The present paper proposes to establish and validate an appropriate threshold for energy classes (high, moderate and low) and for subtidal biological zonation (infralittoral and circalittoral) suitable for EUNIS habitat classification of the Western Iberian coast. Kineticwave-induced energy and the fraction of photosynthetically available light exerted on the marine bottom were respectively assigned to the presence of kelp (Saccorhiza polyschides, Laminaria hyperborea and Laminaria ochroleuca) and seaweed species in general. Both data were statistically described, ordered fromthe largest to the smallest and percentile analyseswere independently performed. The threshold between infralittoral and circalittoral was based on the first quartile while the ‘moderate energy’ class was established between the 12.5 and 87.5 percentiles. To avoid data dependence on sampling locations and assess the confidence interval a bootstrap technique was applied. According to this analysis,more than 75% of seaweeds are present at locations where more than 3.65% of the surface light reaches the sea bottom. The range of energy levels estimated using S. polyschides data, indicate that on the IberianWest coast the ‘moderate energy’ areas are between 0.00303 and 0.04385 N/m2 of wave-induced energy. The lack of agreement between different studies in different regions of Europe suggests the need for more standardization in the future. However, the obtained thresholds in the present study will be very useful in the near future to implement and establish the Iberian EUNIS habitats inventory.

Extracción, identificación y prueba microbiológica del agar extraído de Gracilaria fortissima dawson (rhodophyta, gigartinales, gracilariaceae) (ING)

The red seaweed Gracilaria fortissima colected in the caribean coast of Costa Rica, was studied for the extraction, identification and microbiological performance of the agar contend of this plant, as well as the mineral contend. The research was done focused on the agar quality included pH, % extraction, geling point, fusion point and gel strength, as well as infrared analysis and the performance as a microbiological culture of bacteria and fungus and compared with commercial agar from BDH chemicals.