Although macroalgae usually are inconspicuous on pristine coral reefs, they often thrive on reefs that are subjected to various types of anthropogenic disturbance. This thesis consists of five papers and investigates how biomass and composition of macroalgal communities on coral reefs are affected by regulating factors, such as nutrient availability, herbivory, substrate availability and hydrodynamic forces. In addition, ecological functions and potential impacts of both wild and farmed macroalgal communities are evaluated. Paper I describes a method for using macroalgal tissue nutrient concentrations as bioindicator for nutrient availability, with the possibility to map nutrient loading from larger coastal cities. Papers II and III are manipulative studies comparing top-down and bottom-up regulation of macroalgal communities, where herbivore consumption seems to be the main regulator of biomass whereas nutrient availability mainly influences community composition. Exclosure of large-bodied herbivores had a positive influence on algal biomass in both studies, and during different climatic periods. Paper III also includes the influence of hydrodynamic forces on algal community biomass and structure by comparing a reef crest and a back reef-habitat. Alterations of top-down and bottom-up regulation generally had a stronger effect within the protected back reef-habitat, suggesting that such environments may be more sensitive to anthropogenic influence. Paper IV confirms the general conclusions from papers II and III by studying macroalgal biomass and composition on reef sites with different environmental prerequisites. This study also supports the notion that herbivorous fish can suppress accumulation of macroalgal biomass if substrate availability is low, but not where coral cover is reduced and plenty of substrate is open to macroalgal colonization. The study also found a large temporal variation of macroalgal standing stock and associated nutrients at sites with low top-down regulation. Paper V evaluates potential impacts of seaweed farming on coral reefs and nutrients in the seascape by experimentally studying growth, survival and nutrient binding capacity of Eucheuma denticulatum. This study showed that seaweed farms counteract eutrophication through nutrient extraction and that the risk of farmed algae colonizing local reefs seems to be small as they were rapidly consumed. In conclusion, the studies in this thesis contribute to the understanding of macroalgal regulation and function in tropical seascapes, thereby adding to the knowledge base for coastal management. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript. Paper 5: Manuscript.</p>

Laboratoriniuose tyrimuose koncentruoti trąšos su jūros dumbliais tirpalai turėjo įtakos vasarinių rapsų dygimui, šaknų ir kaleoptilių vystymuisi. Geriausias rezultatas gautas vasarinius rapsus apipurškiant 0,1 proc. vnt. koncentracijos trąšų su jūros dumbliais tirpalu. Šiame variante vasariniai rapsai tolygiausiai sudygo, šaknelių ir kaleoptilių ilgiai buvo vienodžiausi, išsivystė stipri šaknų sistema, kuri leidžia tikėtis gero derlingumo ir atsparesnių augalų blogoms meteorologinėms sąlygoms. Lauko eksperimente trąšos su jūros dumbliais turėjo įtakos vasarinių rapsų derlingumui ir cheminei sudėčiai. Naudojant trąšas su jūros dumbliais vasarinių rapsų derlingumas esmingai padidėjo nuo 0,37 iki 0,42 t ha-1 ir atitinkamai siekė 2,89 ir 2,94 t ha-1. Trąšos su jūros dumbliais taip pat turėjo įtakos vasarinių rapsų riebalų kiekiui. Riebalų kiekio esminis padidėjimas 1,10 proc. vnt. pastebėtas vasarinius rapsus tręšiant Eurofertil plius NP 37 Physio trąšomis prieš sėją, stiebo augimo pradžioje Sulfammo 30 ir butanizacijos tarpsnyje Fertileader Gold trąšomis. Trąšos su jūros dumbliais baltymų ir gliukozinolatų kiekiams esminės įtakos neturėjo. / The laboratory studies of the concentrated fertilizer with seaweed liquid had an impact on oilseed of summer rapeseed germination, root and the development of coleoptiles. The best result was reached by sputtering 0.1 percents units concentration of fertilizer with seaweed on the summer rapeseed. For this fertilizing version the summer rapeseed most germinated, and root and coleoptiles lengths had been the same and it had developed the strong root system, which allowed expecting a good yield and planting more resistant to bad weather conditions. The experiment on field of fertilizer with seaweed had affected the productivity and chemical composition of summer rapeseed. Using the fertilizer with seaweed the productivity of summer rapeseed was substantially increased from 0.37 to 0.42 t ha-1, respectively reached 2.89 and 2.94 t ha-1. The fertilizers with seaweed had also influenced the fat content of summer rapeseed. It was observed the substantial increase of 1.10 percents for unit on the summer rapeseed fertilized with Eurofertil plius NP 37 Physio before sowing, at stage of stem growth time fertilized with Sulfammo 30, at bud formation time with Fertileader Gold. The fertilizers with seaweed hadn’t significant influence to protein and glucosinolate contents.

There is little published information on the distribution, abundance, seasonality and ecological roles of benthic algae on the Great Barrier Reef, although they are of fundamental importance in the ecology of coral reef communities. This study sought to provide information on algal community dynamics in two contrasting reef-flat zones: the live coral and algal turf-dominated outer flat, and the fleshy macroalgaldominated inner flat, at Heron Reef, Great Barrier Reef, Queensland, Australia.Algal community structure on dead coral limestone was studied on the inner and outer reef flat over a three-year period (from May 1986 to February 1989), and algal transplant and grazer exclusion experiments were carried out to determine the role of grazers in producing and maintaining community structure.Changes during ecological succession provide useful information about structuring processes in natural communities. Algal succession was studied using artificial substratum (concrete), because it provided a flat surface which offered logistical advantages over naturally contoured coral substratum, and because previous studies had found that artificial substrata adequately mimicked natural surfaces in marine successional studies. Succession on natural substratum was also studied for comparison, using coral clumps which had died after a bleaching episode in February 1987. The effects of season on algal succession, were investigated using four series of concrete settlement blocks, started in February, May, August and November 1986. The effect of habitat complexity on succession was examined by comparing succession on an exposed concrete surface with succession on a protected concrete surface provided with crevices. Each successional study lasted two years. Cover data for all algal species present on each sample were obtained using a sampling grid of sixty points, and these data were analysed using the non-parametric multivariate analysis program, PRIMER.A total of 105 algal taxa were identified in this study, with 101 of these recorded in the natural reef flat algal community over the three-year period. Fleshy macroalgae were the dominant group on the inner flat (56% cover), and filamentous turf algae iv were dominant on the outer flat (47% cover). The importance of water temperature and grazing intensity in producing the differences in distribution of these two groups of algae on the reef-flat were considered. Temperature was rejected as a factor, because there was virtually no temperature difference between the two sites over a two-year period. Grazing was found to be an important factor, based on a much higher rate of damage to algal transplants on the outer flat than on the inner flat; and on the development of higher algal biomass on caged than on uncaged concrete settlement blocks.Between 58 and 69 algal taxa were recorded in each of the four successional studies on exposed concrete, 78 taxa were recorded on protected concrete, and 85 taxa were recorded on bleached coral. Algal successional trajectories were similar on concrete and on bleached coral, but the rate varied substantially. Succession was fastest on bleached coral, slowest on exposed concrete, and intermediate on protected concrete. On all substrata, at both sites, early dominance by green and brown filamentous algae decreased over time, while the abundance of other forms increased. Dominance in terms of cover by early successional algae (in particular Ectocarpus) was prolonged on concrete in comparison to bleached coral, and this effect was stronger on exposed concrete than on protected concrete.It is hypothesised that inhibition of algal community development on concrete because of a relatively dense growth of early successional algae was due to one or both of two factors: 1. Reduced algal recruitment due to a lower level of substratum heterogeneity; and 2. Reduced grazing by the dominant herbivores in the system, parrotfish (Family Scaridae), because of the hardness of concrete.Succession was faster on the outer flat than on the inner flat. On bleached coral, succession to a community similar to the natural background algal community took between three and nine months on the outer flat, and between one and two years on the inner flat.There was no regular seasonal pattern of change in the reef flat algal community over a three-year period, but during early succession, algal community structure was v strongly influenced by season of initial exposure of the concrete settlement blocks. However, this variability did not persist, and by two years all seasonal series of blocks had similar communities. Six species of algae were fertile within 10 days of establishment, including the common early successional algae Ectocarpus, Enteromorpha and Polysiphonia, illustrating the potential of these species for rapid exploitation of available space.The results of this study provide much needed baseline data on algal community dynamics on the southern Great Barrier Reef, and on succession and community development following a bleaching episode. Given the rapid and dense colonisation of bare substratum by algae, and the very low rate of recruitment by corals, it seems that corals will be at a disadvantage in maintaining their abundance in this reef flat environment if coral bleaching becomes a regular event.

NOBRE, Clareane Avelino Simplicio Nobre. Isolamento, purificação e caracterização parcial da estrutura primária de uma ficobiliproteína da alga marinha vermelha Hypnea musciformis (Wulfen) Lamouroux. 2015. 55 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Engenharia de Pesca, Fortaleza-CE, 2015 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-12T12:16:54Z No. of bitstreams: 1 2015_dis_casnobre.pdf: 1293747 bytes, checksum: ef415f9b1a75c524f87b2a6e87d52332 (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-12T12:17:27Z (GMT) No. of bitstreams: 1 2015_dis_casnobre.pdf: 1293747 bytes, checksum: ef415f9b1a75c524f87b2a6e87d52332 (MD5) / Made available in DSpace on 2016-07-12T12:17:27Z (GMT). No. of bitstreams: 1 2015_dis_casnobre.pdf: 1293747 bytes, checksum: ef415f9b1a75c524f87b2a6e87d52332 (MD5) Previous issue date: 2015 / Seaweeds are a rich reserve of substances with biotechnological interest, among these substances it is possible to highlight the phycobiliproteins. These molecules are water-soluble proteins covalently linked to pigments with linear tetrapyrrole structure called bilin, also known as accessory pigment. Thus, the phycobiliproteins form a complex named phycobilisomes, which is a primary structure of light-gathering, showing photosynthetic function. These structures are found in red and Cyanophyceae seaweed. Phycobiliproteins isolated and purified, when structuraly analised, present subunits characterized as alpha, beta and gamma. These proteins can be used as colorants and stabilizers for food, markers of biomolecules, antioxidants and anti-inflammatory agents. In view of the importance of these molecules, this work aimed to isolate, purify and determine the partial primary structure of the beta chain phycobiliprotein present in the red marine algae Hypnea musciformis. The phycobiliprotein (HMp) was isolated by precipitation of proteins with ammonium sulphate and purified by ion-exchange chromatography. The polyacrylamide gel electrophoresis revealed that the HMp has two chains around 20 kDa and 22 kDa. The 22 kDa chain was excised from the gel and digested with proteolytic enzyme trypsin. The peptides from digestion underwent fragmentation in mass spectrometer. The generated fragments were used for sequencing peptides and have identified in databases. Primers were designed to amplify the beta chain gene bhmp from the genomic DNA of the algae. The amplified gene was sequenced and the tool Phred/Phrap/Consed processed the raw data. The final gene sequence has been translated to obtain the partial primary structure of bHMp. The translation resulted in a sequence with 87 amino acid residues and HMp presented identity with various phycoerythrins of red algae. / As algas marinhas constituem uma rica reserva de substância de interesse biotecnológico, dentre essas substâncias podemos destacar as ficobiliproteínas. Essas moléculas são proteínas solúveis em água ligadas covalentemente a pigmentos com estrutura tetrapirrólica linear denominado bilina, também conhecido como pigmento acessório. Dessa forma, as ficobiliproteínas formam um complexo denominado ficobilissomos, que constitui uma estrutura primordial de captação de luz, apresentando função fotossintética. Essas estruturas são encontradas nas algas marinhas vermelhas e cianofíceas. Ficobiliproteínas isoladas e purificadas, quando caracterizadas, apresentaram subunidades estruturais, como subunidades alfa, beta e gama. Quanto a suas aplicações, essas proteínas foram utilizadas como corantes e estabilizantes de alimentos, marcadores de biomoléculas, agentes antioxidantes e anti-inflamatórios. Tendo em vista a importância destas moléculas, este trabalho teve como objetivo isolar, purificar e determinar a estrutura primária parcial da cadeia beta de uma ficobiliproteína presente na alga marinha vermelha Hypnea musciformis. A ficobiliproteína (HMp) foi isolada através da precipitação do extrato proteico com sulfato de amônio e purificada por cromatografia de troca iônica. A eletroforese em gel de poliacrilamida revelou que a HMp possui duas bandas proteicas em torno de 20 kDa e 22 kDa. A banda de 22 kDa foi excisada do gel e digerida com a enzima proteolítica tripsina. Os peptídeos oriundos da digestão foram submetidos à fragmentação em espectrômetro de massas. Os fragmentos gerados foram utilizados para sequenciar os peptídeos e estes na identificação de proteínas a partir dos bancos de dados existentes. A partir dessas informações, iniciadores foram desenhados para amplificar o gene da cadeia beta da ficobiliproteína de Hypnea musciformis (bhmp) a partir do DNA genômico da alga. O gene amplificado foi sequenciado e os dados brutos foram processados pela ferramenta Phred/Phrap/Consed. A sequência processada foi traduzida para obtenção da estrutura primária parcial de bhmp. A tradução resultou em uma sequência de 87 resíduos de aminoácidos e HMp apresentou identidade com diversas ficoeritrinas de algas vermelhas.

Made available in DSpace on 2014-06-11T19:26:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-09-02Bitstream added on 2014-06-13T20:55:20Z : No. of bitstreams: 1 losi_lc_me_botfca.pdf: 446005 bytes, checksum: 7d7c4bb09f14e32d875db21ebf299ae5 (MD5) / Universidade Estadual Paulista (UNESP) / O presente trabalho teve por objetivo avaliar os efeitos de diferentes doses de extrato de Ascophyllum nodosum na produção de mudas de Eucalytus urograndis e Eucalyptus urophilla durante a fase de enraizamento. Os experimentos foram conduzidos em casa de vegetação, pertencentes às empresas VEC Florestal e Monte Flora, produtoras de microestacas de eucalipto e localizadas na cidade de Bofete, estado de São Paulo. O experimento foi conduzido em quatro fases no período de abril de 2009 a março de 2010, utilizando-se dois clones de E. urograndis (Euca 103 e Euca 105) e um clone de E. urophilla (I144), dois substratos (Brasil Minérios e Carolina Soil) e aplicação de diferentes doses de extrato de A. nodosum (EAN) 0, 0,5, 1, 1,5, 2, 3, 4, 8 e 16 mL L-1. O delineamento experimental adotado foi em blocos ao acaso com 4 repetições e 86 plantas por parcela. As aplicações do extrato de A. nodosum ocorreram aos 0, 7, 14, 21 e 28 dias após o estaqueamento variando conforme a fase. Foram avaliados a massa fresca de raiz (MFR), massa seca de raiz (MSR), comprimento de raiz (CR), massa fresca de parte aérea (MFA), massa seca de parte aérea (MAS), diâmetro do caule (D) e quantidade de raiz (Q) aos 30 e 45 2 dias após dias após o estaqueamento e análise química da planta. As avaliações permitiram observar que houve resposta diferenciada do EAN em relação aos substratos e ao material genético. O tratamento na dose de 3 mL de EAN para o clone I144 proporcionou melhor enraizamento das microestacas. Observou-se também que a forma de aplicação do produto interfere nos resultados / The objective of this work was to evaluate the effects of different rates of extract of Ascophyllum nodosum in the production of microshoots Eucalyptus urograndis and Eucalyptus urophilla during the rooting. The experiments were conducted in a greenhouse belonging to companies VEC Florestal and Monte Flora, two producers of microcuttings eucalyptus and located in the city of Bofete, State of São Paulo. The experiment was conducted in four phases between April 2009 and March 2010, using two clones of E. urograndis (Euca 103 e Euca 105) and one clone of E. urophilla (I144), two substrates (Brasil Minérios and Carolina Soil) and application of different rates of extract A. nodosum (EAN) 0, 0,5, 1, 1,5, 2, 3, 4, 8, 16 mL L-1. The experimental design used was randomized blocks with four replications and 86 plants per plot. The applications of the extract A. nodosum occur at 0, 7, 14, 21 and 28 days after the cutting. It were evaluated root fresh weight (MFR), root dry mass (MSR), root length (CR), fresh weight of shoot (MFA), shoot dry mass (MAS), stem diameter (D) and quality of root (Q) at 30 and 45 days after the cutting. The evaluations allowed to note that there was differential response of EAN in relation to substrates and genetic material, The dose of 3 mL of ean for clone I144 provided better microcuttings rooting. It was also observed that the way of the product application affect the results

Made available in DSpace on 2014-12-17T14:02:02Z (GMT). No. of bitstreams: 1 VanessaMFC.pdf: 1244215 bytes, checksum: 2d84570105aa3f980f60bc5ac24ab268 (MD5) Previous issue date: 2006-09-11 / Dentre as macroalgas capazes de absorver altas concentra??es de N e P dissolvidos na ?gua, destaca-se a Chlorophyta Ulva lactuca, bastante adapt?vel e resistente ?s adversidades ambientais, como grandes varia??es de temperatura, salinidade, mat?ria org?nica e metais pesados. Trata-se tamb?m de uma esp?cie bastante comum nas ?reas intertidais do litoral norte-riograndense. Devido a suas caracter?sticas ecol?gicas, fisiol?gicas e nutricionais, foi avaliado nesse estudo, o seu potencial como biofiltro na redu??o de NH4+, NO3- e PO4-2, tanto em condi??es controladas como tamb?m em um viveiro de camar?o. No experimento laboratorial, foram utilizados quatro aqu?rios de vidro de 30 x 20 x 20cm com 10L de ?gua, sendo tr?s aqu?rios experimentais contendo 20g de U. lactuca e um controle. O acr?scimo de biomassa foi de 2,92g (22,92 ? 6,29g; p < 0,05) em rela??o ao in?culo inicial de 20g, sob temperatura (28,50 ? 0,58?C), salinidade (35,00 ? 0,00 ), pH (8,26 ? 0,02) e luz constante (250 &#956;mol.m2s-1). O crescimento positivo (1,78 ? 4,38%dia-1; p < 0,05), juntamente com a alta efici?ncia de absor??o de am?nio (83%; p < 0,001), nitrato (83%; p < 0,001) e ortofosfato (53%; p < 0,001), demonstrou que, nessas condi??es, a Ulva lactuca absorveu os nutrientes e aumentou sua biomassa. J? no experimento de campo, realizado na fazenda TECNAR?O, situada no munic?pio de Arez/RN (06? 11 40 Latitude Sul, e 35? 09 37 Longitude Oeste), foram utilizadas tr?s gaiolas de PVC, posicionadas a 12cm da superf?cie da ?gua, cada uma com dimens?es 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 ?s condi??es controladas, demonstrando a adaptabilidade da esp?cie em condi??es ambientais vari?veis, onde, apesar da temperatura pouco vari?vel (27,45 ? 0,64?C), houve progressiva diminui??o de salinidade (25 - 15 ), devido ao per?odo de fortes chuvas (34,70 ? 23,78mm). Somado a isso, foram observados v?rios fatores biol?gicos interferindo no viveiro, como a presen?a de ep?fitas, organismos endof?ticos, fouling e a herbivoria por parte dos pr?prios camar?es. Houve aumento nas concentra??es de NH4+ (4,36 ? 1,69 &#956;mol.L-1), NO3- (0,17 ? 0,25&#956;mol.L-1) e PO4-2 (0,41 ? 0,13&#956;mol.L-1), coincidindo com o crescimento da esp?cie at? a terceira semana. Todos os par?metros ambientais analisados, assim como a biomassa e a Taxa de Crescimento Relativo (TCR), obtidos no campo, apresentaram varia??es altamente significativas (p < 0,001). As correla??es 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 esp?cie ? capaz de ter um crescimento satisfat?rio nas condi??es eutr?ficas de um viveiro de camar?o, sendo poss?vel seu uso como biofiltro.

Made available in DSpace on 2014-12-17T14:03:35Z (GMT). No. of bitstreams: 1 CelinaMPGD_TESE.pdf: 1816885 bytes, checksum: 1e39ca48cae61af571d696fef8f71be4 (MD5) Previous issue date: 2012-10-15 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The present study examines the chemical composition and their effects on free radicals, inflammation, angiogenesis, coagulation, VEGF effects and cellular proliferation of a polysaccharides from alga Sargassum vulgare. The sulfated polysaccharide was extracted from brown seaweed by proteolysis with enzymes maxataze. The presence of proteins and sugars were observed in crude polysaccharides. Fractionation of this crude extract was made with growing concentration of acetone (0.3-1.5 v) and produced four groups of polysaccharides. Anionic polysaccharides from brown seaweed Sargassum vulgare, SV1and PSV1 were fractionated (SV1) and purified (PSV1), and displayed with high total sugars and sulfate content and very low level of protein. This fucan SV1 contains low levels of protein and high carbohydrate and sulfate content. This polysaccharides prolonged activated partial thromboplastin time (aPTT) at 50 &#956;g (>240 s). SV1 was found to have no effect on prothrombin time (PT), corresponding to the extrinsic pathway of coagulation. SV1 exhibits high antithrombotic action in vivo, with a concentration ten times higher than heparin. Polysaccharides from S. vulgare promoted direct inhibition enzymatic activity of thrombin and stimulated enzymatic activity of FXa. SV1 showed optimal inhibitory activity of thrombin (50.2?0.28%) at a concentration of 25 &#956;g/mL. Its antioxidant action on scavenging radicals by DPPH was (22%), indicating the polymer has no cytotoxic action (hemolytic) on ABO and Rh blood types in different erythrocyte groups and displays strong anti-inflammatory action on all concentrations tested in the carrageenan-induced paw edema model, demonstrated by reduced edema and cellular infiltration. Angiogenesis is a dynamic process of proliferation and differentiation. It requires endothelial proliferation, migration, and tube formation. In this context, endothelial cells are a preferred target for several studies and therapies. The antiangiogenic efficacy of polysaccharides was examined in vivo in the chick chorioallantoic membrane (CAM) model by using fertilized eggs. Decreases in the density of the capillaries were assessed and scored. The results showed that SV1 and PSV1 have an inhibitory effect on angiogenesis. These results were also confirmed by inhibition tubulogenesis in rabbit aorta endothelial cell (RAEC) in matrigel. These compounds were assessed in Apoptosis assay (Annexin V - FITC / PI) and cell viability by MTT assay of RAEC. These polysaccharides do not affect the viability and do not have apoptotic or necrotic action. RAEC cell when incubated with SV1 and PSV1showed inhibition of VEGF secretion, observed when compounds were incubated at 25, 50 and 100 &#956;g/&#956;L. The VEGF secretion with the RAEC cell line for 24 h, was more effective for PSV1 at 50 &#956;g/&#956;L(71.4%) than SV1 100 &#956;g/&#956;L (75.9%). SV1 and PSV1 had an antiproliferative action (47%) against tumor cell line HeLa. Our results indicate that these sulfated polysaccharides have antiangiogenic and antitumoral actions / O presente estudo analisa a composi??o qu?mica e seus efeitos sobre os radicais livres, inflama??o, angiog?nese, coagula??o, VEGF e prolifera??o celular dos polissacar?deos de uma alga Sargassum vulgare. O polissac?rido sulfatado foi extra?do a partir de algas marrons por prote?lise com a enzima maxataze. A presen?a de prote?nas e a??cares foram observados no cru de polissacarideos. Fracionamento do o extrato bruto foi feito com concentra??es crescente de acetona (0,3-1,5 v), produzindo quatro grupos de polissacarideos. Estes compostos ani?nicos da alga S. vulgare, foram fracionados (SV1) e purificados (PSV1) exibindo com alta a??cares totais e sulfatecontent e n?vel muito baixo de prote?nas.A fucana SV1 cont?m baixos n?veis de prote?na e de hidratos de carbono e alto teor de sulfato. Este polissacar?deos prolongou o tempo de tromboplastina parcial activada (aPTT) a 50 ug (>240 s). n?o foi observado qualquer efeito de SV1 sobre o tempo de protrombina (PT), que corresponde a via extr?nseca da coagula??o. SV1 exibiu alta a??o antitromb?tica in vivo, com uma concentra??o 10 vezes maior do que a heparina. SV1 promoveu a actividade de inibi??o enzim?tica direta da trombina e estimulou a atividade enzim?tica do FXa. Mostrou tamb?m, atividade inibidora optima de trombina (50,2 ? 0,28%) a uma concentra??o de 25 ug / mL. A sua ac??o anti-oxidante de radicais scavenging por DPPH foi de (22%), indicando que o pol?mero n?o tem qualquer a??o citot?xica (hemol?tica) em tipos de sangue ABO e Rh, em diferentes grupos de eritr?citos e exibindo alta a??o anti-inflamat?ria em edema de pata de ratos Wistar em todas as concentra??es testadas induzida por carragenina. Tal processo foi demonstrado por edema e infiltra??o celular. A angiogenese ? um processo din?mico de prolifera??o e diferencia??o. Ele requer prolifera??o endotelial, migra??o, e a forma??o do tubo. Neste contexto, as c?lulas endoteliais s?o um alvo preferido para muitos estudos e terapias. A efic?cia antiangiogenico de polissacar?deos foi examinada in vivo na membrana corioalant?ica pinto (CAM) usando-se ovos fertilizados. Diminui??es na densidade dos capilares foram avaliados e pontuados. Os resultados mostraram que SV1 e PSV1 tem um efeito inibidor da angiogenese. Estes resultados foram tamb?m confirmados por tubulogenesis inibi??o na c?lula endotelial da aorta de coelho (RAEC) em matrigel. C?lulas RAEC quando foram incubadas com SV1and PSV1 demonstraram inibi??o da secre??o de VEGF, a 25, 50 e 100 ug/mL. A secre??o de VEGF com a linha de c?lulas RAEC durante 24 h, foi mais eficaz para PSV1 a 50 ug / mL (71,4%) do que SV1 100 ug / mL (75,9%). SV1 e PSV1 posuiram uma ac??o antiproliferativa (47%) contra as c?lulas tumorais tipo HeLa. Estes compostos foram avaliados tamb?m, no ensaio de apoptose (anexina V - FITC / PI) e a viabilidade celular pelo ensaio de MTT de RAEC. Estes polissacar?deos n?o afetaram a viabilidade e n?o tiveram a??o apopt?tica ou necr?tica. Nossos resultados indicam que estes polissacar?deos sulfatados t?m a??es antiangiog?nica e antitumoral e constituem um importante alvo biol?gico e farmacol?gico

Orientador: Roberto Lyra Villas Bôas / Banca: Antonio de Pádua Sousa / Banca: Ana Cláudia Pacheco Santos / Resumo: O presente trabalho teve por objetivo avaliar os efeitos de diferentes doses de extrato de Ascophyllum nodosum na produção de mudas de Eucalytus urograndis e Eucalyptus urophilla durante a fase de enraizamento. Os experimentos foram conduzidos em casa de vegetação, pertencentes às empresas VEC Florestal e Monte Flora, produtoras de microestacas de eucalipto e localizadas na cidade de Bofete, estado de São Paulo. O experimento foi conduzido em quatro fases no período de abril de 2009 a março de 2010, utilizando-se dois clones de E. urograndis (Euca 103 e Euca 105) e um clone de E. urophilla (I144), dois substratos (Brasil Minérios e Carolina Soil) e aplicação de diferentes doses de extrato de A. nodosum (EAN) 0, 0,5, 1, 1,5, 2, 3, 4, 8 e 16 mL L-1. O delineamento experimental adotado foi em blocos ao acaso com 4 repetições e 86 plantas por parcela. As aplicações do extrato de A. nodosum ocorreram aos 0, 7, 14, 21 e 28 dias após o estaqueamento variando conforme a fase. Foram avaliados a massa fresca de raiz (MFR), massa seca de raiz (MSR), comprimento de raiz (CR), massa fresca de parte aérea (MFA), massa seca de parte aérea (MAS), diâmetro do caule (D) e quantidade de raiz (Q) aos 30 e 45 2 dias após dias após o estaqueamento e análise química da planta. As avaliações permitiram observar que houve resposta diferenciada do EAN em relação aos substratos e ao material genético. O tratamento na dose de 3 mL de EAN para o clone I144 proporcionou melhor enraizamento das microestacas. Observou-se também que a forma de aplicação do produto interfere nos resultados / Abstract: The objective of this work was to evaluate the effects of different rates of extract of Ascophyllum nodosum in the production of microshoots Eucalyptus urograndis and Eucalyptus urophilla during the rooting. The experiments were conducted in a greenhouse belonging to companies VEC Florestal and Monte Flora, two producers of microcuttings eucalyptus and located in the city of Bofete, State of São Paulo. The experiment was conducted in four phases between April 2009 and March 2010, using two clones of E. urograndis (Euca 103 e Euca 105) and one clone of E. urophilla (I144), two substrates (Brasil Minérios and Carolina Soil) and application of different rates of extract A. nodosum (EAN) 0, 0,5, 1, 1,5, 2, 3, 4, 8, 16 mL L-1. The experimental design used was randomized blocks with four replications and 86 plants per plot. The applications of the extract A. nodosum occur at 0, 7, 14, 21 and 28 days after the cutting. It were evaluated root fresh weight (MFR), root dry mass (MSR), root length (CR), fresh weight of shoot (MFA), shoot dry mass (MAS), stem diameter (D) and quality of root (Q) at 30 and 45 days after the cutting. The evaluations allowed to note that there was differential response of EAN in relation to substrates and genetic material, The dose of 3 mL of ean for clone I144 provided better microcuttings rooting. It was also observed that the way of the product application affect the results / Mestre

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / As algas marinhas sÃo fontes abundantes de polissacarÃdeos sulfatados com vÃrias atividades biolÃgicas dessa forma suas biomolÃculas sÃo de grande interesse comercial principalmente nas indÃstrias farmacÃutica e alimentÃcia No presente trabalho investigou-se uma fraÃÃo polissacarÃdica sulfatada obtida da alga marinha verde Caulerpa racemosa (CrII) quanto aos seus efeitos na nocicepÃÃo e inflamaÃÃo Inicialmente a CrII (1,0 mg/kg i.v.) foi avaliada quanto a toxicidade em camundongos utilizando um modelo por dose repetida (sete dias) ApÃs o perÃodo experimental a CrII mostrou-se atÃxica A atividade antinociceptiva foi avaliada atravÃs dos ensaios de contorÃÃes abdominais induzidas por Ãcido acÃtico teste da formalina e da placa quente Camundongos Swiss machos foram tratados com CrII (0,01 0,1 e 1,0 mg/kg i.v.) 30 min antes de receber os estÃmulos de dor (Ãcido acÃtico 0,8% ou formalina 2%) ou exposiÃÃo ao calor Nos ensaios de contorÃÃes abdominais induzidas por Ãcido acÃtico CrII reduziu significativamente o nÃmero de contorÃÃes abdominais e no teste da formalina CrII tambÃm foi capaz de reduzir o tempo de lambedura da pata na segunda fase do experimento Em relaÃÃo ao estÃmulo tÃrmico CrII nÃo foi capaz de prolongar o tempo de reaÃÃo dos animais. Com relaÃÃo aos efeitos na inflamaÃÃo CrII (0,01 0,1 e 1,0 mg/kg i.v.) foi avaliada quanto seu potencial anti-inflamatÃrio utilizando-se ratos Wistar Foram utilizados ensaios de migraÃÃo celular para a cavidade peritoneal induzida por carragenana (Cg - 700 Âg/cavidade) de edema de pata induzido por Cg (700 Âg/pata) ou dextrana (300 Âg/pata) Com relaÃÃo a migraÃÃo celular para a cavidade peritoneal ocorreu uma reduÃÃo significativa de neutrÃfilos em todos os grupos tratados com a CrII No ensaio de edema de pata induzido por Cg ou dextrana CrII tambÃm foi capaz de reduzir significativamente o edema em todas as doses utilizadas O efeito anti-inflamatÃrio da CrII foi confirmado atravÃs da reduÃÃo dos nÃveis teciduais da mieloperoxidase do tecido das patas nos grupos com Cg AlÃm disso foi realizado um ensaio de inibiÃÃo da enzima Hemo-oxygenase-1 (HO-1) utilizando-se o inibidor ZnPP IX com o intuito de avaliar se o efeito anti-inflamatÃrio da CrII estava relacionado com a expressÃo dessa enzima Os resultados demonstraram que a inibiÃÃo da via HO-1 està associada à inibiÃÃo da resposta anti-inflamatÃria da CrII No intuito de avaliar se CrII tambÃm possui efeito prÃ-inflamatÃrio CrII (0,01 0,1 e 1,0 mg/kg i.pl) foi injetada na pata de ratos Wistar onde o resultado obtido demonstrou um processo inflamatÃrio apenas para animais que receberam a dose de 1,0 mg/kg Portanto os possÃveis mediadores que poderiam estar envolvidos com o processo inflamatÃrio da CrII foram avaliados utilizando-se o ensaio de edema de pata induzido por CrII Os resultados demonstraram que a aÃÃo prÃ-inflamatÃria da CrII pode estar relacionada com a liberaÃÃo de mediadores provindos da via da ciclooxigenase (COX-2 prostaglandinas e tromboxanos) A CrII quando utilizada como agente anti-inflamatÃrio nÃo foi capaz de inibir seu efeito prÃ- inflamatÃrio no ensaio de edema de pata em nenhuma das doses utilizadas (0,01 0,1 e 1,0 mg/kg i.v.) confirmando que sua aÃÃo anti-inflamatÃria està relacionada com a via da HO-1 / Marine algae are abundant sources of sulfated polysaccharides with various biological activities thereby altering its biomolecules are great of commercial interest especially in the pharmaceutical and food industries In this study we investigated a sulfated polysaccharide obtained from the green seaweed Caulerpa racemosa (CrII) as to its effects on nociception and inflammation Initially the CrII (1.0 mg/kg, i.v.) was evaluated for toxicity in mice using a repeated dose model (seven days) After the trial period the CrII proved to be nontoxic The antinociceptive activity was evaluated by the writings of abdominal constrictions induced by acetic acid formalin test and hot plate Swiss male mice were treated with CrII (0.01 0.1 and 1.0 mg/kg i.v.) 30 min before receiving pain stimuli (acetic acid 0.8% or 2% formalin) or heat exposure In trials of abdominal constrictions induced by acetic acid and the formalin test CrII significantly reduced the number of abdominal writhing and paw licking time in the second phase of the experiment respectively In relation to thermal stimulation CrII was unable to prolong the reaction time of animals With respect to effects on inflammation CrII (0.01 0.1 and 1.0 mg/kg i.v.) was evaluated for its potential antiinflammatory using Wistar rats We used assays of cell migration into the peritoneal cavity induced by carrageenan (Cg - 700 mg/cavity) paw edema induced by Cg (700 Âg/paw) or dextran (300 Âg/paw)With respect to cell migration into the peritoneal cavity there was a significant reduction of neutrophils in all groups treated with the CrII In the test paw edema induced by dextran or Cg CrII was also able to significantly reduce the swelling at all doses used The anti-inflammatory effect of CrII was confirmed by reducing the tissue levels of myeloperoxidase in the tissue of the paws Cg groups In addition we performed an enzyme inhibition assay Hemo-oxygenase-1 (HO-1) using the inhibitor ZnPP IX in order to assess whether the anti-inflammatory effect of CrII was related to the expression of this enzyme The results showed that the inhibition of the HO-1 is associated with inhibition of the inflammatory response of CrII In order to assess whether CrII also has pro-inflammatory effect CrII (0.01 0.1 and 1.0 mg/kg i.pl) was injected into the paw of the rats where the result demonstrated a process inflammation only for animals which received the dose of 1.0 mg/kg Therefore the potential mediators that could be involved in the inflammatory process of CrII were evaluated using the test paw edema induced by CrII The results showed that the pro-inflammatory effect of CrII may be related to the release of mediators emanating from the path of cyclooxygenase (COX-2 prostaglandins and thromboxanes) The CrII when used as anti-inflammatory agent was unable to inhibit pro-inflammatory effect in the paw edema test in any of the doses (0.01 0.1 and 1.0 mg/kg i.v.) confirming that its anti-inflammatory effect is related to HO-1

Algas marinhas sÃo seres classificados no reino protista, predominantemente aquÃticos e com grande capacidade adaptativa. Encontradas em todas as regiÃes do planeta, as algas desempenham importante papel ecolÃgico e econÃmico, representando um grande potencial de biorrecursos. A alga marinha vermelha Gracilaria birdiae, coletada durante doze meses, foi submetida à extraÃÃo aquosa a 100 ÂC de polissacarÃdeos e apresentou rendimento superior a 50 %, com exceÃÃo nos meses de fevereiro, setembro e novembro de 2007 ( 49,5%, 41,8% e 49,4% respectativamente). O teor de sulfato foi obtido por microanÃlise e nÃo variou significativamente ao longo dos meses analisados, apresentando valor mÃdio de 1,35%. O grau de sulfataÃÃo mÃdio encontrado foi de 0,15. O conteÃdo protÃico nas amostras tambÃm nÃo apresentou variaÃÃo considerÃvel, tendo mÃdia de 0,5%, entretanto, no mÃs de abril o conteÃdo protÃico aumentou cerca de 3 vezes do valor mÃdio encontrado. As doze amostras foram caracterizadas quimicamente por cromatografia de permeaÃÃo em gel (GPC), espectroscopia na regiÃo do infravermelho (FT-IR) e por ressonÃncia magnÃtica nuclear (RMN). Os cromatogramas de GPC das amostras apresentaram semelhanÃas e caracterÃsticas tÃpicas de polissacarÃdeos das algas vermelhas. Em geral, foram observados um pico principal, em aproximadamente 7,58 mL e um ombro em aproximadamente 9,2 mL de volume de eluiÃÃo, com exceÃÃo dos meses de agosto, outubro e novembro de 2007, que apresentaram cromatogramas com pico principal em aproximadamente 8,27 mL de volume de eluiÃÃo, detectados pelo Ãndice de refraÃÃo. Os polissacarÃdeos investigados apresentaram altas massas molares , variando de 0,11 x 104 a 8,51 x 105 g/mol. Os espectros de infravermelho das doze amostras de polissacarÃdeos apresentaram perfis semelhantes, com as bandas caracterÃsticas de agarocolÃides, mais especificamente, do tipo agarana. Espectros de RMN foram empregados para a caracterizaÃÃo estrutural das amostras ao longo do ano e, revelaram que a estrutura do polissacarÃdeo à composta por segmentos de (1&#8594; 3)-&#946;-D-galactopiranosil ligada a (1&#8594;4)-&#945;-L-anidrogalactopiranosil. O estudo da variaÃÃo sazonal da composiÃÃo centesimal, em geral, nÃo apresentou variaÃÃo acentuada. Com relaÃÃo à quantidade de proteÃnas solÃveis e a atividade hemaglutinante no extrato protÃico, o estudo da variaÃÃo sazonal revelou uma pequena variaÃÃo no teor de proteÃnas solÃveis, com valor mÃximo (0,94 mgP/mL) em novembro/06 e mÃnimo no mÃs de dezembro/06 (0,39 mgP/mL). Em geral, a atividade hemaglutinante observada em Gracilaria birdiae à fraca, porÃm, ela esteve presente em concentraÃÃes variÃveis ao longo de todo o perÃodo estudado, com exceÃÃo no mÃs de dezembro/06, independente do conteÃdo protÃico total e solÃvel encontrado. / Seaweeds are classified in the kingdom protist, predominantly aquatic and with great adaptive capacity. Found in all regions of the planet, algae play an important role ecological and economic, representing a great potential for bioresources. The red seaweed Gracilaria birdiae, collected during twelves months, was submitted to aqueous extraction of polysaccharides at 100ÂC, showed yield exceeding 50 %, except in February, September and November 2007 (49,5%, 41,8% and 49,4% respectively). The sulfate content was obtained by microanalysis and it not vary significantly over the months examined averaging 1,35%. The sulfate degree was 0.15. The protein content in the samples also showed no considerable variation, with average of 0,5%, howevwr in April, the protein content increased about 3 times the average found. The twelve samples were chemically characterized by Gel Permeation Chromatography (GPC), Infrared (FT-IR) and Nuclear Magnetic Resonance (NMR) spectroscopy. The GPC chromatograms of the samples showed similarity and typical characteristics of polysaccharides from red algae. In general, we observed a main peak, at approximately 9,2 mL volume of eluition, with the exception of August, October and November 2007, which showed chromatograms with main peak at approximately 8,7 mL volume of eluition, detected by a refractive index. The polysaccharides investigated showed hight molecular weight, ranging from 0.11 x 104 and 8.51 x 105 g/mol. The infrared spectra of the twelve samples of polysaccharides presented similar profiles, with bands characteristics of agarocolloids, more specifically agaran-like, with evidence of protein. NMR were employed to the structural characterization of the samples throughout the year and revealed that the structure of polysaccharides is composed by (1&#8594;3)-&#946;-D-galactopyranosyl linkage to (1&#8594;4)-&#945;-L-anhydrogalactopyranosyl segments. The study of seasonal variation in proximate composition, in general, showed no marked variation. Regarding the amount of soluble protein in the crude extract, the study of seasonal variation showed a small variation in the content of soluble protein, with maximum value (0,94 mgP/mL) in November/06 and minimum (0,39 mgP/mL) in December/06. The hemagglutinating activity during the period of one year, except in December/06.