Estudos realizados em 1979 por Kelecom e Kanengiesser mostraram que extratos brutos obtidos a partir de amostras da esponja marinha Aplysina fulva não possuíam derivados bromados. Estes foram resultados inesperados, uma vez que esponjas pertencentes à ordem Verongida são conhecidos produtores de metabólitos derivados da dibromotirosina. O presente projeto teve como meta a investigação química de extratos obtidos de duas amostras de A. fulva, sendo uma coletada em São Sebastião (SP) e a outra em Angra dos Reis (RJ), objetivando verificar a ocorrência de derivados da bromotirosina e uma possível variabilidade química dependendo da distribuição geográfica das esponjas. Sete derivados da dibromotirosina foram isolados das duas amostras de Aplysina fulva: quatro compostos da amostra de Angra dos Reis e três da de São Sebastião. As estruturas dos compostos foram identificadas por análises espectroscópicas (como IV, UV, RMN 1H, RMN 13C, HMQC, COSY, HMBC) e de espectrometria de massas, e ainda por comparação com dados da literatura. Os resultados obtidos confirmaram que os derivados da dibromotirosina são bons marcadores quimiotaxonômicos para as esponjas da Ordem Verongida. Além disso, a variabilidade química observada para a A. fulva parece ser influência de fatores abióticos e bióticos como sazonalidade, disponibilidade de nutrientes, ou associação com diferentes micro-organismos. / Studies developed in 1979 by Kelecom and Kanengiesser showed that crude extracts obtained from samples of the marine sponge Aplysina fulva were devoid of bromotyrosine derivatives. These results were rather unexpected, since sponges belonging to the Order Verongida are well-known producers of bromotyrosine-derived metabolites. The present project aimed the chemical investigation of crude extracts obtainded from two samples of A. fulva, one collected at São Sebastião (SP) and the second from Angra dos Reis (RJ), in order to verify the occurrence of bromotyrosine derivatives and a possible chemical variability depending on the geographical distribution of the sponge. Seven bromoyrosine derivatives have been isolated from the two samples of A. fulva: four compounds from the Angra dos Reis sample and three from the São Sebastião sample. The structures of the compounds have been established by spectroscopic analysis, (including IV, UV, RMN 1H, RMN 13C, HMQC, COSY, HMBC) and mass spectrometry, as well as by comparison with literature data. The results obtained confirmed that bromotyrosine derivatives are good chemotaxonomic markers for sponges of the Order Verongida. Moreover, it appears that chemical variability of A. fulva may be influenced by abiotic and biotic factors, such as seasonality, nutrients availability, or association with distinct micro-organisms.

Estudos realizados em 1979 por Kelecom e Kanengiesser mostraram que extratos brutos obtidos a partir de amostras da esponja marinha Aplysina fulva não possuíam derivados bromados. Estes foram resultados inesperados, uma vez que esponjas pertencentes à ordem Verongida são conhecidos produtores de metabólitos derivados da dibromotirosina. O presente projeto teve como meta a investigação química de extratos obtidos de duas amostras de A. fulva, sendo uma coletada em São Sebastião (SP) e a outra em Angra dos Reis (RJ), objetivando verificar a ocorrência de derivados da bromotirosina e uma possível variabilidade química dependendo da distribuição geográfica das esponjas. Sete derivados da dibromotirosina foram isolados das duas amostras de Aplysina fulva: quatro compostos da amostra de Angra dos Reis e três da de São Sebastião. As estruturas dos compostos foram identificadas por análises espectroscópicas (como IV, UV, RMN 1H, RMN 13C, HMQC, COSY, HMBC) e de espectrometria de massas, e ainda por comparação com dados da literatura. Os resultados obtidos confirmaram que os derivados da dibromotirosina são bons marcadores quimiotaxonômicos para as esponjas da Ordem Verongida. Além disso, a variabilidade química observada para a A. fulva parece ser influência de fatores abióticos e bióticos como sazonalidade, disponibilidade de nutrientes, ou associação com diferentes micro-organismos. / Studies developed in 1979 by Kelecom and Kanengiesser showed that crude extracts obtained from samples of the marine sponge Aplysina fulva were devoid of bromotyrosine derivatives. These results were rather unexpected, since sponges belonging to the Order Verongida are well-known producers of bromotyrosine-derived metabolites. The present project aimed the chemical investigation of crude extracts obtainded from two samples of A. fulva, one collected at São Sebastião (SP) and the second from Angra dos Reis (RJ), in order to verify the occurrence of bromotyrosine derivatives and a possible chemical variability depending on the geographical distribution of the sponge. Seven bromoyrosine derivatives have been isolated from the two samples of A. fulva: four compounds from the Angra dos Reis sample and three from the São Sebastião sample. The structures of the compounds have been established by spectroscopic analysis, (including IV, UV, RMN 1H, RMN 13C, HMQC, COSY, HMBC) and mass spectrometry, as well as by comparison with literature data. The results obtained confirmed that bromotyrosine derivatives are good chemotaxonomic markers for sponges of the Order Verongida. Moreover, it appears that chemical variability of A. fulva may be influenced by abiotic and biotic factors, such as seasonality, nutrients availability, or association with distinct micro-organisms.

Made available in DSpace on 2015-04-01T14:16:06Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1615747 bytes, checksum: 2bbe77aa39b7f33d5f7b0781f235bb2d (MD5) Previous issue date: 2014-08-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A new lectin was purified and characterized from marine sponge Aplysina fulva (AFL). The crude extract was obtained with Tris HCl 0.1 M pH 7.4 containing NaCl 0.15 M buffer. Haemagglutinating activity detection and soluble protein measurement were performed with the crude extract and after each purification step. Isolation of AFL was performed by affinity chromatography in Sepharose CL 4B column, and the retained peak was applied in a DEAE Sephacel ion exchange chromatography column. Analysis of DEAE Sephacel retained peak on polyacrylamide gel electrophoresis (SDS-PAGE) showed the presence of a single band with approximately 27.5 kDa. Under non-denaturating conditions it shows the presence of a single 57 kDa band. The novel lectin showed large amount of hydrophobic amino acids, high temperatures resistance and better hemagglutination activity in the range of neutral to alkaline pHs. AFL was able to inhibit the growth of strains of P. aeruginosa, C. albicans and C. tropicalis. The lectin did not show toxic effects on cells of breast cancer, but had agglutination effect on Leishmania brasiliensis promastigotes by recognizing galactose residues on its surface. / Uma nova lectina foi purificada e caracterizada a partir da esponja marinha Aplysina fulva (AFL). O extrato bruto foi produzido a partir da solubilização de proteínas em solução tampão Tris HCl 0,1 M pH 7,4 NaCl 0,15 M, sendo este utilizado na realização dos ensaios de atividade hemaglutinante e dosagem de proteínas pelo método de Bradford. O primeiro passo de purificação de AFL foi por meio de cromatografia de afinidade em coluna de Sepharose CL 4B. O pico não retido foi eluido com o mesmo tampão de extração e o pico retido foi eluido com tampão Glicina 0,1 M pH 2,6 NaCl 0,15 M. O mesmo foi dialisado, liofilizado e ressuspendido em Tris HCl 0,025 M pH 7,6 e submetido a cromatografia de troca iônica em DEAE Sephacel. A análise do pico retido da DEAE Sephacel por eletroforese nativa em gel de poliacrilamida (PAGE) mostrou a presença de uma única banda com aproximadamente 57 kDa, como também uma única banda com aproximadamente em 27,5 kDa em presença de SDS (PAGE-SDS). A nova lectina apresenta uma grande quantidade de aminoácidos hidrofóbicos, mostrando-se resistente a altas temperaturas e com melhor atividade hemaglutinante na faixa de pH neutro a alcalino. AFL foi capaz de inibir o crescimento de cepas de P. aeruginosa, C. albicans e C. tropicalis. A nova lectina não apresentou efeitos tóxicos para células de câncer de mama, entretanto mostrou-se hábil em aglutinar formas promastigotas de Leishmania brasiliensis através do reconhecimento de galactose em sua superfície.

A common injury type associated with vessel groundings on coral reefs is generation of coral rubble. As a substrate rubble is dynamic and can be mobilized by normal wave agitation, causing secondary damage to surviving corals and preventing coral larvae and invertebrates from surviving. Typical rubble management options include offsite removal and onsite stabilization using artificial materials. Sponges show promise as a restoration tool for rubble stabilization, especially sponges that are known to have rapid regeneration rates, low mortality, and the capability to quickly form attachment points to natural and artificial substrates. As an effective restoration tool a sustainable source of sponge is required in order to minimize impact on natural populations. The purpose of this project is to evaluate the feasibility of sponge propagation for use as a restoration material following groundings on coral reefs in Biscayne National Park, FL. If small sponge fragments could be collected from donor sponges and propagated on artificial growout structures (i.e. nurseries) a sustainable source of sponge fragments could be maintained for future restoration activities. If feasible, this technique would create a continually growing sponge source that would require a single harvest, reducing pressure on the local sponge populations. Fragments of four species of native sponges, Aplysina fulva¸ Amphimedon compressa, Aplysina cauliformis, and Iotrochota birotulata, were harvested and attached to artificial growout structures composed of PVC rods and cement paver tiles. Fragment necrosis, attachment, sponge retention (persistence throughout the study), complexity, and growth rates for all four species were recorded monthly for one year. Two different PVC types were utilized to compare attachment material effects on growth. Donor sponges were monitored to assess recovery and growth after harvesting, and intact sponges in the natural population were also monitored as a control. Transplanted fragments had the highest retention of any treatment (96-100% of fragments remained), growth rates ranging from 105% to 424%, low necrosis rates, and all but one species increased in complexity. There was no significant difference in sponge growth on different PVC types or between sites. Within one month all donor wounds inflicted by harvest were completely healed, and by the end of the study donors had significantly higher growth rates than the control treatment, ranging from 400% to 781% of initial sponge volume. Donor sponges also had low necrosis rates, but were less complex than controls. This is a promising, practical and low cost method, and based on these results, is a feasible way of producing sponges for us in restoration, with little negative impact on the natural populations.