Thesis (M.Sc.)--Victoria University of Wellington, 2008. / Includes bibliographical references.

Thesis (M.S.)--University of Alabama at Birmingham, 2007. / Description based on contents viewed May 30, 2008; title from title screen. Includes bibliographical references (p. 31-43).

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-07-14T14:49:52Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertacao Ludimila (completa).pdf: 4542374 bytes, checksum: ae089eb3590a6dac6965ff1e1218a752 (MD5) / Made available in DSpace on 2016-07-14T14:49:52Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertacao Ludimila (completa).pdf: 4542374 bytes, checksum: ae089eb3590a6dac6965ff1e1218a752 (MD5) Previous issue date: 2015-07-31 / CAPEs / Corvoheteromeyenia Ezcurra de Drago, 1979 é um gênero endêmico da Região Neotropical, criado para albergar: C. australis (Bonetto & Ezcurra de Drago, 1966) e C. heterosclera (Ezcurra de Drago, 1974). No entanto, devido à confusão na descrição original de C. australis e C. heterosclera, não era clara a distinção entre as espécies. Adicionalmente, o material tipo dessas espécies não foi depositado nas coleções indicadas nos trabalhos originais. Sendo assim, no intuito de elucidar o status específico destes materiais, o presente trabalho apresenta a revisão do gênero e a designação dos neótipos. Um total de 41 espécimes foram analisados e identificados como sendo Corvoheteromeyenia. Baseado na bibliografia e no material examinado foi realizada a redescrição das espécies C. australis e C. heterosclera. A microsclera acantóxea foi apontada como o caráter distintivo entre ambas as espécies, presente apenas em C. heterosclera. Além destas duas espécies, foi descrita C. sanidastosclera Pinheiro, Silva & Calheira, no prelo, que difere das demais por apresentar apenas gemosclera sanidáster. Este trabalho também testou se a fisiologia da eclosão da gêmula poderia influenciar na distribuição das esponjas de águas continentais. Até o presente, apenas a morfologia gemular estava sendo considerados para entender o padrão de distribuição das espécies. Neste trabalho, escolhemos duas espécies (Heteromeyenia cristalina Batista, Volkmer-Ribeiro & Melão, 2007 e Radiospongilla inesi Nicacio & Pinheiro, 2011) para testar se a distribuição da espécie está associada à capacidade de eclosão da gêmula em diferentes tipos de ambientes. Cinco tratamentos foram realizados (T): T1 (água do local da coleta das esponjas); T2 (água do Rio Pirangi); T3 (água da Lagoa Araraquara); T4 (água mineral) e T5 (T1 + gêmulas dessecadas). Para cada tratamento, foram utilizadas 30 gêmulas com três réplicas. As gêmulas foram observadas diariamente, durante 30 dias. Para a análise dos dados foi realizada ANOVA e teste Tukey. O resultado apresentou que as gêmulas de ambas as espécies submetidas ao T3 não eclodiram, provavelmente em virtude da maior condutividade da água. Houve diferença significativa entre os tratamentos (T1, T2, T4) e entre as espécies (F2;12=77,2). A comparação entre T1 e T5 apresentou diferença entre os tratamentos e as espécies (F1;8=27,5), com alta significância para ambos resultados (P<0.001). Foi constatado que R. inesi apresentou um percentual de eclosão elevado nos tratamentos analisados, enquanto que H. cristalina, teve percentual de eclosão elevado apenas na água de sua própria localidade de coleta. Adicionalmente foi observado o desenvolvimento de esponjas em ambas as espécies. / Corvoheteromeyenia Ezcurra de Drago, 1979 is endemic from the Neotropical Region and was created to comprise C. australis (Bonetto & Ezcurra de Drago, 1966) and C. heterosclera (Ezcurra de Drago, 1974). However, due to confusion in the original description of C. australis and C. heterosclera, the distinction between them was not well-established. Additionally, the type material from these species was not deposited in the collections indicated at the original descriptions. Thus, in order to elucidate their specific status, this paper presents a review of the genus and the designation of both neotypes. A total of 41 specimens were analyzed and identified as belonging to Corvoheteromeyenia. Based on the literature and additional specimens, the redescription of C. australis and C. heterosclera is presented. The microsclera acanthoxea was stated as the distinctive character of both, present only in C. heterosclera. Corvoheteromeyenia sanidastosclera Pinheiro, Silva & Calheira, in press, was also described and is distinct from congeners by the presence of gemmoscleres sanidaster. This study also tested whether the physiology of gemmules hatching could affect the distribution of freshwater sponges. Until the present, only the morphology of the gemmule was being considered to understand the pattern of distribution of species. In this work we choose two species (Heteromeyenia cristalina Batista, Volkmer-Ribeiro & Melão, 2007 e Radiospongilla inesi Nicacio & Pinheiro, 2011) to test whether the distribution is associated with the capacity of gemmules hatching in different environments. Five treatments were done (T): T1 (water of sponges site collected); T2 (Pirangi River water); T3 (Araraquara Pond water); T4 (mineral water) and T5 (T1 + drying gemmules). For each treatment 30 gemmules with three replicates were used. The gemmules were observed daily during 30 days. Data analysis was performed using ANOVA and Tukey test. Results showed that gemmules from both species submitted to T3 have not hatched, probably due to its higher conductivity. There was a significant difference between treatments (T1, T2, T4) and between species (F2, 12 = 77.2). Comparison between T1 and T5 showed differences between treatments and species (F1, 8 = 27.5), with high significance for both results (P <0.001). It was found that R. inesi presented a high percentage of hatching at all treatments while H. cristalina had high percentage of hatching only at treatment with water from its natural habitat. Additionally, the development of sponges was observed from both species.

Neste trabalho descreve-se o estudo química dos extratos de quatro esponjas e dois fungos de origem marinha oriundos da costa do Brasil. Os extratos de três esponjas (Petromica ciocalyptoides, Topsentia ophiraphidites e Callyspongia sp.) apresentaram atividade inibitória à enzima adenosina fosforribosil transferase de Leishmania tarentolae. A partir desses extratos foram isolados 4 compostos. O trissulafato de halistanol, isolado das esponjas P. ciocalyptoides e T. ophiraphidites, e o ilhabelanol, ilhabreno e isoakaterpina, isolados da esponja Callyspongia sp. A partir do extrato bruto da esponja Axinella cf corrugata foram isolados dois derivados cumarínicos, provavelmente artefatos de isolamento do ácido 4-esculetínico, o qual é inédito como produto natural. O extrato bruto da esponja Axinella cf. corrugata apresentou atividade citotóxica, mas os compostos puros não apresentaram esta atividade. Os dois compostos puros foram testados ainda quanto sua atividade contra o vírus da SARS, na qual o éster etílico do ácido 4-esculetínico se apresentou ativo. A partir de dois extratos oriundos do fungo Beauveria felina, isolado da alga marinha Caulerpa sp, foram isoladas 17 frações puras que após diversas análises foram agrupadas em seis compostos conhecidos na literatura: a (Phe3, N-Val5) destruxina B, a cloroidrina da destruxina E, a roseotoxina B, a roseocardina, a isariina e a isariina B. Além disso, foram isolados dois compostos inéditos, a pseudodestruxina C e a cloloidrina Beta-Me-Pro da destruxina E. Os extratos brutos de Beauveria felina apresentaram atividades em bioensaios de atividade antituberculose e de citotoxicidade em linhagem de células de câncer. Os compostos puros avaliados no bioensaio antituberculose não foram ativos. Somente o composto roseotoxina B apresentou atividade citotóxica in vitro para quatro linhagens de células: mama, cólon, sistema nervoso e leucemia. / In this work we report the chemical investigation of bioactive crude extracts obtained from four sponges and two fungal strains of marine origin. The crude extracts of three sponges species (Petromica ciocalyptoides, Topsentia ophiraphidites and Callyspongia sp.) displayed inhibitory activity towards the enzyme adenine fosforribosyl transferase of Leishmania tarentolae (L-APRT). Four compounds have been isolated from these extracts: the known halistanol sulfate was isolated of sponges P. ciocalyptoides and T. ophiraphidites, while the novel ilhabelanol, ilhabrene and isoakaterpin have been isolated from the sponge Callyspongia sp. All compounds exhibited inhibition of L-APRT at micro M concentrations. Two coumarin derivatives have been isolated from the crude extract of the sponge Axinella cf. corrugata, probably as artifacts of isolation: esculetin-4-carboxylic acid methyl ester and esculetin-4-carboxylic acid ethyl ester. While the crude extract of the sponge Axinella cf. corrugata presented cytotoxic activity, the pure compounds were inactive in these assays. The esculetin-4-carboxylic acid ethyl ester was found to be an in vitro inhibitor of SARS virus. The crude extract obtained of a marine-derived Beauveria felina strain, isolated from the alga Caulerpa sp., displayed antituberculosis activity against Mycobacterium tuberculosis H37Rv and cytotoxic activity against MCF-7 (breast), HCT-8 (colon) and B16 (murine melanoma) cancer cell lines. Chemical fractionation of the crude extract led to the isolation of two new cyclodepsipeptides pseudodestruxin C and [Beta-Me-Pro] destruxin E chlorohydrin, and of the known destruxin E chlorohydrin, [Phe3, N-Me-Val5] destruxin B, roseotoxin B, roseocardin, isariin and isariin B. The depsipeptides [Phe3, NMe- Val5] destruxin B and rosetoxin B, have been tested against M. tuberculosis H37 Rv and in cytotoxicity bioassays against SF 295 (human CNS) MDA-MB435 (human breast) HCT8 (colon) and HL60 (leukemia) cancer cell lines. Only roseotoxin B displayed moderate cytotoxicity against the cancer cell lines.

An in depth chemical investigation of the major and minor pyrroloiminoquinone metabolites produced by four species of endemic South African Latrunculid sponges, collected from Algoa Bay and the Tsitsikamma Marine Reserve off the south eastern coast of South Africa, yielded eleven new and twelve known pyrroloiminoquinone metabolites. The structures of the new metabolites were determined using standard spectroscopic techniques. Tsitsikamma pedunculata was shown to contain 7,8-dehydro-3-dihydro-discorhabdin C (2.1), 14-bromo-7,8-dehydro-3-dihydro-discorhabdin C (2.2), discorhabdin S (2.3), 14-bromo-1-hydroxy-discorhabdin S (2.4), 1-bromo-2-hydroxy-4-debromo-discorhabdin S (2.5), and 2,4-debromo-3-dihydro-discorhabdin C (2.6), together with the known compounds 14-bromo-discorhabdin C (1.51), 14-bromo-3-dihydro-discorhabdin C (1.52) and 3-dihydro-discorhabdin C. The metabolites from T. pedunculata were characterised by the presence of a reduced C-3 carbonyl and bromination at C-14. Compounds isolated from a second Latrunculid sponge, Latrunculia lorii, ranged from a substituted bicyclic pyrrolecarboxylic acid, makaluvic acid A (1.47), to the simple tricyclic known pyrroloiminoquinones makaluvamine C (1.33) and damirone B (1.20) and the more complex discorhabdin D type metabolites, discorhabdin M (3.2), 1-amino discorhabdin D (3.3), 1-methoxy discorhabdin D (3.4) and 1-alanyl discorhabdin D (3.5). Discorhabdin G* (3.1) was also isolated and characterised. This is the first reported occurrence of the known compounds 1.20, 1.33 and 1.47 in a Latrunculia sponge. Discorhabdin and bis-pyrroloiminoquinone type compounds predominated in Tsitsikamma favus. Three known, tsitsikammamines A (1.71) and B (1.72), 1.52, and five new pyrroloiminoquinones, tsitsikammamine N-oxime (4.1), tsitsikammamine B N-oxime (4.2), 2.1, 2.4 and 2.6, were isolated from this sponge. A fourth Latrunculid sponge (Strongylodesma sp.) yielded three known compounds, discorhabdins A (1.57), D (1.61) and 1.53, and one new pyrroloiminoquinone 3.3. The dual role of these metabolites as cytotoxic agents and pigments resulted in an attempt to relate the photochemical properties of these metabolites to their cytotoxicity. The pyrroloiminoquinone metabolites studied exhibited moderate singlet oxygen quantum yields, while three compounds (1.57, 4.1 and 4.2) were shown to be capable of producing radicals at a wavelength of 532 nm. The possibility of a correlation between the electrochemical properties and anti-cancer (HCT-116) activity of selected pyrroloiminoquinones was explored. A study of the oesophageal and ovarian cytotoxicities of two pyrroloiminoquinones (1.57 and 1.72), together with an investigation into the intercalation and topoisomerase I inhibitory activity of the bis-pyrroloiminoquinones (1.71, 1.72, 4.1 and 4.2), are presented.

Sponges play an important role in many marine habitats and are crucial for maintaining the deep-sea marine ecosystems. However, there is a knowledge gap in the field of sponge biodiversity due to insufficient exploration of the deep-sea, and the probability of finding new species is fairly high. The deep waters of the Indian and Pacific Oceans are particularly poorly explored, and their sponge diversity is barely known. During the KANADEEP 2 expedition in the south of New Caledonia a large collection of Demospongiae and Hexactinellida was gathered to assess the sponge biodiversity. In this project 110 Demosponges were investigated morphologically and identified to the order level. Twenty-one of these were selected for further identification and description in terms of shape and geometry of spicules using light microscopy. The DNA was extracted from eight specimens for the confirmation of new species. Amplification of CO1 Folmer fragment was conducted using PCR. The resulting PCR-products were analyzed using gel electrophoresis and DNA-sequencing. The twenty-one specimens were assigned to the genera Tethya, Stupenda and Geodia belonging to the order Tetractinellida, which was found to be the dominant order in the deep waters in the New Caledonia region. There are potentially six new species amongst the studied specimens. However, it has to be confirmed with molecular analysis of specific markers. The morphological analysis of 21 specimens collected during the KANADEEP 2 expedition demonstrated that 17 specimens belonged to Geodia, two specimens to Tethya, and two specimens to Stupenda.

This study of the distribution of the fresh-water sponge, Meyenia fluviatilis, at the Lincoln Beach area of Utah Lake involved the following: First, sampling of the area for the presence or absence of the species. Second, sampling at two separate areas at different depth intervals to determine if the number of sponge colonies and colony size were in any way related to depth. Finally, monthly sampling at one area to determine when gemmules of M. fluviatilis were present in the sponge colonies. Meyenia fluviatilis was found to be commonly distributed in 1-2 m of water on the lower surface of the larger rocks at 30 transects approximately 100 m apart. At the 1-2 m depth interval, the largest number of colonies (36 colonies per 50 rocks at area A and 69 colonies at area 8) were recorded. The average colony size was also greatest at this same depth (21.5 cm for area A and 26 cm for area 8). The lowest monthly percent of gemmules observed at area 8 occurred in June (22 %), while August was the highest observation with 70 %. Gemmules were observed every month and no sponge colony disintegration was observed during any month.

No description available.

No description available.

Algoa Bay Latrunculid sponges are well known for their production of cytotoxic pyrroloiminoquinones with speculation that these secondary metabolites may have a microbial origin. This study describes a thorough investigation into the bacterial community associated with Tsitsikamma favus, Tsitsikamma scurra a newly described Latrunculia sp. and a yellow encrusting sponge associated with T. scurra. Molecular and chemical characterisation were used in conjunction with traditional taxonomy in identification of the sponge specimens. The 28S rRNA and COX1 analysis confirmed the traditional taxonomy with T. favus and T. scurra being very closely related. Chemical analysis revealed that T. favus and T. scurra shared the discorhabdins 2,4-debromo-3-dihydrodiscorhabdin C, 7,8-dehydro-3-dihydrodiscorhabdin C and 14-bromo-1-hydroxy-discorhabdin V in common with each other and Tsitsikamma pedunculata indicating that these pyrroloiminoquinones are common to Tsitsikamma sponges in general. The bacterial community associated with T. favus was explored using 16S rRNA molecular techniques including DGGE, clonal libraries of full length 16S rRNA genes, as well as 454 pyrosequencing. DGGE analysis revealed that the bacterial community associated with T. favus appeared to be highly conserved, which was confirmed by both the clone library and 454 pyrosequencing, with the Betaproteobacteria as the most dominant class. Further exploration into T. favus, as well as T. scurra, Latrunculia sp. and the yellow encrusting sponge indicated that the bacterial populations associated with each of these sponge species were conserved and species specific. OTU analysis to the species level revealed that T. favus and T. scurra shared an abundant Spirochaete species in common while the most abundant species in the Latrunculia sp. and the yellow encrusting sponge belonged to the class Betaproteobacteria. The exclusivity of the tsitsikammamines to T. favus precipitated attempts to culture the T. favus associated bacteria, with a focus on the dominant betaproteobacterium as indicated by the 16S rRNA clone library. Actinobacteria associated with the Algoa Bay sponge specimens were also cultured and the actinobacterial isolates were sent for screening against Mycobacterium aurum with two Kocuria kristinae isolates and a Streptomyces albdioflavus isolate showing good antimycobacterial activity.