Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Lectinas sÃo (glico)proteÃnas que podem reconhecer e se ligar reversivelmente a carboidratos ou a outras substÃncias derivadas de aÃÃcares. SÃo encontradas nos reinos vegetal e animal. Hypnea cervicornis à uma espÃcie de alga marinha vermelha encontrada no nordeste do Brasil da qual foi isolada a lectina H. cervicornis aglutinina (HCA), um polipeptideo que contÃm 90 resÃduos de aminoÃcidos (9193 Da) que se ligam especificamente a glicoproteÃnas do tipo mucina. O objetivo do presente trabalho foi estudar as atividades antiinflamatÃria e antinociceptiva, ainda nÃo exploradas, da HCA. Foram utilizados camundongos Swiss albinos (20-35g) ou ratos Wistar (150-250g). Para o estudo da atividade antiinflamatÃria da HCA, os ratos foram injetados e.v. somente com a lectina nas doses de 10-2; 10-1; 1 e 10 mg/Kg (peritonite) ou a dose de 1 mg/Kg em associaÃÃo com 5 mg/mL de mucina (1 mg/Kg). ApÃs esse procedimento, os animais foram avaliados nos modelo de peritonite induzida Cg (quimiotÃtico direto) ou por fMLP (quimiotÃtico indireto). HCA (10-1; 1 e 10 mg/Kg) inibiu a peritonite induzida por Cg e por fMLP. Observamos que na peritonite induzida por Cg: A administraÃÃo conjunta de HCA (1 mg/Kg) com mucina inibiu sua atividade antiinflamatÃria (p<0,05), sugerindo participaÃÃo do sÃtio lectÃnico nesta atividade; o aquecimento de HCA a 100oC, por 10 minutos, inibiu seu efeito antiinflamatÃrio, indicando a importÃncia de sua estrutura; a microscopia intravital mostrou que a HCA (10 mg/Kg) reduziu o rolamento e adesÃo de leucÃcitos; a lectina induziu a produÃÃo de Ãxido nÃtrico (NO, reaÃÃo de Griess) no soro; Aminoguanidina e L-Nitro-Arginina inibiram a atividade da HCA; HCA nÃo reduziu as concentraÃÃes de citocinas TNF-&#945; e IL-1 no peritÃnio. A pressÃo arterial e o leucograma dos animais nÃo foram alterados pela administraÃÃo de HCA. A lectina nÃo age diretamente sobre neutrÃfilos, pois nÃo inibiu a quimiotaxia in vitro induzida por agente quimiotÃtico (MIP-2). No edema de pata induzido por Dextrana ou Cg, HCA (1 mg/Kg) inibiu significativamente (p<0,05) apenas o edema de pata induzido por Cg. O efeito antiedematogÃnico da HCA foi prevenido pela administraÃÃo conjunta com mucina. HCA reduziu as contorÃÃes induzidas por Ãcido acÃtico e seu efeito antinociceptivo foi prevenido pela administraÃÃo conjunta com mucina. HCA diminuiu somente a segunda fase do teste da formalina. No teste da placa quente, HCA nÃo apresentou efeito. A lectina tambÃm nÃo alterou a atividade locomotora. A toxicidade sub-crÃnica foi avaliada pelo tratamento de ratos com HCA (1mg/kg), durante sete dias consecutivos, atravÃs de vÃrios parÃmetros: funÃÃes do rim (peso Ãmido, dosagem de creatinina e urÃia) e do fÃgado (peso Ãmido, avaliaÃÃo da cinÃtica da aspartato amino transaminase e alanina amino transaminase), coraÃÃo (peso Ãmido), estÃmago (peso Ãmido e avaliaÃÃo visual de possÃveis lesÃes), variaÃÃo de massa corporal dos animais tratados e leucograma. Os resultados obtidos nÃo mostraram qualquer alteraÃÃo dos parÃmetros avaliados, demonstrando que a HCA nÃo apresenta nenhuma toxicidade nos animais. Em conjunto, esses dados revelam que a HCA apresenta atividade antiinflamatÃria e antinociceptiva que supostamente podem ser explicadas por bloqueio direto competitivo com carboidratos ligantes de selectinas e/ou indireto, via produÃÃo de NO, que diminuem o rolamento e adesÃo de leucÃcitos sobre o endotÃlio. / Lectins are (glyco)proteins that can recognize and reversibly bind to carbohydrates or other substances derived from sugars. They are found throughout animal and vegetal kingdoms. Hypnea cervicornis is a species of marine red algae found in Northeast of Brazil from which was isolated the lectin Hypnea cervicornis aglutinin (HCA), a polypeptide containing 90 amino acid residues (9193 Da) that binds especifically to mucin-type glycoprotein. The objective of this work was to study the anti-inflammatory and anti-nociceptive activities of HCA not yet known. Albine Swiss mice (20-35g) and Wistar rats (150-250g) were used. For the study of HCA anti-inflammatory activity, the rats were injected (i.v.) with lectin alone in the doses of 10-2; 10-1; 1 e 10 mg/Kg (peritonitis) or a dose of 1 mg/Kg associated to 5 mg/mL of mucin (1 mg/Kg). Following this procedure, the animals were evaluated in the peritonitis model induced by Cg (direct neutrophil chemoatractant) or by fMLP (indirect neutrophil chemoatractant). HCA (10-1; 1 and 10 mg/Kg) inhibited Cg- and fMLP-induced peritonitis. In Cg-induced peritonitis it was observed that: the administration of HCA (1mg/Kg) in association with mucin inhibited its anti-inflammatory activity (p<0.05), suggesting the participation of the lectin site on this mechanism; heating HCA at 100oC, for 10 minutes, inhibited its anti-inflammatory effect, indicating the importance of its structure for that mechanism; the intravital microscopy showed that HCA reduced the rolling and adhesion of the leucocytes; the lectin induced the production of nitric oxide (NO Griess reaction) in serum; aminoguanidine and L-Nitro-arginin inhibited the HCA activity; HCA did not reduce the concentrations of TNF-&#945; and IL-1 in the peritoneum. The arterial pressure and the leukogram of the animals were not altered by the administration of HCA. The lectin does not act directly on neutrophils because it did not inhibit the in vitro chemotaxis induced by MIP-2, a chemotactic agent. In the carragenan (Cg)- or dextran-induced paw edema, HCA (1 mg/Kg) inhibited significantly (p<0.05) only the Cg-induced paw edema. The antiedematogenic effect of HCA was prevented when it was administered in association with mucin. HCA reduced the writhes induced by acetic acid and its antinociceptive effect was prevented by the association with mucin. HCA only decreased the second phase of formalin test. In the hot plate, HCA did not show any effects. The lectin did not alter the motor coordination. The HCA subcronic toxicity was evaluated treating rats with a dose of 1mg/Kg during seven days and analys of several parameters: kidney functions (wet weight, creatinin and urea serum concentrations), liver functions (wet weight, kinetic of serum aspartate aminotransferase and alanine aminotransferase), heart (wet weight) stomach (wet weight and visual evaluation of possible lesions), variation of body weight of treated animals and leukogram. The results showed no alterations of the evaluated parameters suggesting that HCA seems to be well tolerated by animals. The results taken together reveal that HCA presents anti-inflammatory and nociceptive activities which could be explained by direct competitive blockage with a common selectin carbohydrate ligand and/or indirectly, via NO production, which decrease of rolling and adhesion of leukocytes to endothelium.

Les espèces coralliennes Acropora palmata (Lamarck, 1816) et A. cervicornis (Lamarck, 1816) ont un rôle majeur en termes de bio-construction et structuration des récifs dans la Caraïbe. Cependant, les populations de ces deux espèces sont en déclin et elles sont maintenant classées en danger critique d’extinction par l’Union Internationale de Conservation de la Nature (UICN). De récentes études principalement menées sur les récifs de la Floride et des Grandes Antilles se sont ainsi intéressé à la structure et à la dynamique des populations de ces espèces, mais le statut génétique des populations des Petites Antilles est moins connu.Le présent sujet a pour but d’étudier l’état des populations des coraux Acropora palmata et A. cervicornis sur les récifs de Guadeloupe et des Petites Antilles et d’élucider les mécanismes qui président à la survie de ces espèces et au recrutement des jeunes coraux. / In the Caribbean, Acropora palmata (Lamarck 1816) and A. cervicornis (Lamarck 1816) are major coral species for reef building. Since the 80’s, these species populations are decreasing and are now classified as critically endangered species by the International Union for Conservation of Nature (IUCN). Recent studies, mainly conducted on the reefs of Florida and the Greater Antilles concerned the structure and the dynamics of Acropora populations, while the genetic status of the populations in the Lesser Antilles remains less studied. The purpose of the study is to determine the status of the coral populations of Acropora palmata and A. cervicornis on reefs of Guadeloupe and Lesser Antilles and to elucidate the mechanisms governing the survival of these species and the recruitment of young corals.

Made available in DSpace on 2014-12-17T14:03:35Z (GMT). No. of bitstreams: 1 RafaelBGC_DISSERT.pdf: 2123374 bytes, checksum: 0195cd61affe048d50e8378ceba77a87 (MD5) Previous issue date: 2010-10-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / In the present study, extracts rich-sulfated polysaccharides were obtained from three different species of Dictyotales (a class of brown macroalgae): Canistrocarpus cervicornis, Dictyota mertensii and Dictyopteris delicatula and their anticoagulant and antioxidant activities were evaluated. All extracts showed anticoagulant activity on aPTT assay, but not on PT assay. Extracts also exhibited total antioxidant activity, superoxide radical scavenging capacity and ferric chelating property. The extract from C. cervicornis showed the best results and was choose to have their sulfated polysaccharides fractioned and subsequently analysed. Thus, six fractions (CC-0.3, CC-0.5, CC-0.7, CC-1.0, CC-1.2 and CC-2.0) were obtained by proteolysis followed by sequential acetone precipitation. Agarose gel eletrophoresis stained with blue toluidine, confirmed the presence of sulfated polysaccharides in all fractions. Chemical analyses showed that all fractions presented heterofucans mainly constitued by fucose, galactose, glucuronic acid and sulfate. Any fraction changed the PT. However, all fractions were able to double the aPTT on a dose-dependent manner. CC- 0.3, CC-0.5, CC-0.7 and CC-1.0 needed only 0.100 mg/mL to double the aPTT, result only 1.25 times higher than the Clexane? (0.080 mg/mL), a commercial low molecular heparin. The heterofucans presented appreciable total antioxidant capacity, low capacity on scavenging hydroxyl radical and good efficiency on scavenging superoxide radicals (except CC-1.0). CC-1.2 showed 43.1 % on superoxide radical scavenging. This result was higher than that showed by the same concentration of gallic acid (41.8 %), a known antioxidant. Furthermore, the heterofucans showed excelent activity on ferrous chelating activity (except CC-0.3). CC-0.5, CC-0.7 and CC-1.0 showed the highest activities with 47.0 % of ferrous chelating activity, a result 2.0 times lesser than that exhibited by the same concentration of EDTA. These results clearly indicated the beneficial effects of heterofucans extracted from C. cervicornis as potential anticoagulant and antioxidant agents. However additional steps of purification, structural studies, besides in vivo experiments are needed for these fucans may be used as therapeutic agents / No presente estudo, extratos brutos ricos em polissacar?deos sulfatados foram obtidos a partir de tr?s esp?cies de Dictyotales (uma ordem de macroalgas marrons): Canistrocarpus cervicornis, Dictyota mertensii e Dictyopteris delicatula, e suas atividades anticoagulante e antioxidante foram avaliadas. Todos os extratos apresentaram atividade anticoagulante frente ao ensaio de aPTT, mas n?o sobre o ensaio de PT. Os extratos tamb?m exibiram atividade antioxidante total, capacidade em sequestrar radicais super?xido e propriedade de quelar ferro. O extrato obtido a partir de C. cervicornis apresentou os melhores resultados e foi escolhido para ter seus polissacar?deos sulfatados fracionados e subsequentemente analisados. Deste modo, seis fra??es (CC-0.3, CC-0.5, CC-0.7, CC-1.0, CC-1.2 e CC-2.0) foram obtidas por prote?lise seguida de precipita??o sequencial com acetona. Eletroforese em gel de agarose corada com azul de toluidina comprovou a presen?a de polissacar?deos sulfatados em todas as fra??es. As an?lises qu?micas mostraram que todas as fra??es apresentam heterofucanas constitu?das principalmente por fucose, galactose, ?cido glucur?nico e sulfato. Nenhuma fra??o alterou o PT. Entretanto todas as fra??es foram capazes de dobrar o aPTT de uma maneira dose dependente. As fra??es, CC-0.3, CC-0.5, CC-0.7 e CC-1.0, precisaram de apenas 0,100 mg/mL para dobrar o tempo de coagula??o, concentra??o que ? apenas 1,25 vezes maior do que aquela utilizada com Clexane? (heparina de baixo peso molecular) para se obter o mesmo efeito. As heterofucanas apresentaram apreci?vel capacidade antioxidante total, baixa capacidade em sequestrar radicais hidroxila e uma boa efici?ncia em sequestrar radicais super?xido (exceto CC-1.0). CC-1.2 mostrou uma capacidade de sequestrar 43,1 % dos radicais super?xido. Este resultado foi maior do que o apresentado pela mesma concentra??o de ?cido g?lico (41,8 %), um antioxidante conhecido. Al?m disso, as heterofucanas mostraram uma excelente atividade em quelar ferro (exceto CC-0.3). CC-0.5, CC-0.7 e CC-1.0 apresentaram as maiores atividades com 47,0 % de quela??o f?rrica, um resultado 2 vezes menor do que o exibido pela mesma concentra??o de EDTA. Estes resultados indicaram claramente os efeitos ben?ficos de heterofucanas extra?das de C. cervicornis como potenciais agentes anticoagulante e antioxidante. Entretanto, passos adicionais de purifica??o, estudos estruturais, al?m de experimentos in vivo, s?o necess?rios para que estas fucanas possam vir a ser utilizadas como agentes terap?uticos

Made available in DSpace on 2014-12-17T14:02:03Z (GMT). No. of bitstreams: 1 MarcellaAAC.pdf: 1596539 bytes, checksum: 311fd18dd515bf47c2eb0788bcbd609a (MD5) Previous issue date: 2007-03-07 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / 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 ?mol.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 ?mol g-1 dry wt h-1) and P-PO43- in low concentrations (Ks = 5 ?mol.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 / O objetivo deste estudo foi avaliar o crescimento da macroalga Gracilaria cervicornis em um viveiro de camar?o (Litopenaeus vannamei) e determinar a efici?ncia de absor??o e os par?metros cin?ticos (Vmax, Ks e Vmax:Ks) desta esp?cie para os nutrientes N-NH4+, N-NO3- e P-PO4-3, visando a sua utiliza??o como biorremediadora de ambientes eutrofizados. Para este estudo, foram desenvolvidos dois experimentos (campo e laborat?rio). No estudo em campo, a macroalga foi examinada em rela??o ao crescimento e a biomassa. No experimento em laborat?rio, a efici?ncia de absor??o de G. cervicornis foi medida atrav?s do monitoramento das concentra??es dos tr?s nutrientes (N-NO3-, N-NH4+ e P-PO4-3) durante 5 horas e os par?metros cin?ticos foram determinados atrav?s da f?rmula de Michaelis-Menten. Os resultados obtidos neste estudo demonstraram que G. cervicornis se beneficiou dos nutrientes dispon?veis no viveiro, aumentando 52,4% do valor de sua biomassa ap?s 30 dias de cultivo. Neste estudo foi constatado que a variabilidade da biomassa p?de ser explicada atrav?s da salinidade, disponibilidade de luz (transpar?ncia e concentra??o de part?culas s?lidas em suspens?o) e concentra??es do N-NO3- no ambiente (r-Pearson= 0,76; p<0,05). Quanto ao experimento em laborat?rio, foi observado que a efici?ncia de absor??o da macroalga estudada foi maior nos tratamentos com menor concentra??o (5 ?mol.L-1), sendo constatada uma redu??o de at? 85,3%, 97,5% e 81,2% do N-NH4+, N-NO3- e P-PO43-, respectivamente. Quanto aos par?metros cin?ticos, G. cervicornis apresentou uma maior habilidade em absorver o N-NH4+ em altas concentra??es (Vmax = 158,5 ?mol g-1 dry wt h-1) e o P-PO43- em baixas concentra??es (Ks = 5 ?mol.L-1 e Vmax:Ks = 10,3). Os resultados deste estudo mostram que G. cervicornis pode ser cultivada em viveiros de camar?o e que apresenta uma boa capacidade de absor??o dos nutrientes testados, podendo desta forma, ser uma candidata promissora como biorremediadora de efluentes de camar?o

This thesis is composed of two articles. The first is an analysis of long-term survival among A. cervicornis outplants in the Florida Keys, from 2007 to the present. The second is a review of literature that informs coral restoration and guides both restoration practitioners and coral researchers towards greater effectiveness in outplant survival and understanding biological processes involved in restoration. In the first article, despite promising initial evidence of outplant survival and health, the long-term results, using Weibull survival analysis, are discouraging with almost all out planted corals over an 8 year long study exhibiting steep declines in percent live tissue and survival between three and five years. Not only is this 3-5 year collapse apparent in all the outplanted cohorts, but the evidence is highly significant that the length of outplant survival is decreasing with each passing year (diminished resilience). These findings suggest that some shared, likely environmental factor, is increasingly impacting all outplants. Further, no cohorts appear to adapt to the environmental conditions in which these declines are occurring (diminished adaptive capacity), a trend that would be evident if their declines slowed or reversed and Weibull beta-parameterization would show. The second article, reviews several areas of recent study which offer avenues for future research: these include, ecological history and biogeography, developmental pathways of colonial form and function, polarity and symmetry, genetics, wound healing, fecundity, reproduction, sexual maturity and community interactions. The thesis concludes with questions for further research and understanding in the field of coral restoration.

Submitted by Clebson Anjos (clebson.leandro54@gmail.com) on 2016-03-29T18:44:13Z No. of bitstreams: 1 arquivototal.pdf: 4614259 bytes, checksum: e4765945b6c16211d7771a473b84ad25 (MD5) / Made available in DSpace on 2016-03-29T18:44:13Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 4614259 bytes, checksum: e4765945b6c16211d7771a473b84ad25 (MD5) Previous issue date: 2013-12-05 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Marine biodiversity is presented as a promising source of new substances with high biological potential. Among the many organisms that inhabit the marine environment, we stress the brown algae belonging to the family Dictyotaceae. Genus Canistrocarpus was recently established, and some algae Dictyota formerly were reclassified as Canistrocarpus. This work describes the results of the phycochemical study of Canistrocarpus cervicornis (formerly classified as Dictyota pulchella), and the evaluation of the phycochemical profile of C. cervicornis collected on the coast of Paraíba and the coast of Rio de Janeiro, which contributed to the correct name of Dictyota pulchella as Canistrocarpus cervicornis. Alga samples were collected at Bessa Beach (coordinates 07º04'01 '' S and 34º49'35 '' W), city of João Pessoa (PB) in August 2010 and December 2012. The material was dried at room temperature and extraction was performed with CH2Cl2 : MeOH (2:1). The material was dried at room temperature in fresh air and extraction was performed with CH2Cl2: MeOH (2:1). The extract obtained the first collection was subjected to vacuum filtration on silica gel with solvent hexane, ethyl acetate and methanol in increasing gradient of polarity. The fraction hexane : ethyl acetate (85:15) was subjected to column chromatography yielded one new natural product, the dolastane diterpene 4,7-diacetoxy dichotenone A. The extract resulting from the second collection was divided in two parts. The first part was subjected to column chromatography resulted in the isolation of a known dolastane diterpene, 4-acetoxy-14-hidroxydolast-1(15),7,9-triene. A second aliquot of the extract was acetylated and subjected to column chromatography, which led to the isolation of three diterpenes, one dolastane unpublished in the literature, 4-desoxy-amijidictyol and two known secodolastanes, isolinearol acetate and indicol. Chemical constituents isolated were identified by spectroscopic methods 1H and 13C-1D and 2D NMR, optical rotation and comparison with literature data. This study contributed to the chemotaxonomy of the tribe Dictyoteae, especially C. cervicornis. / A biodiversidade marinha apresenta-se como uma fonte promissora de novas substâncias com grande potencial biológico. Dentre os diversos organismos que habitam o ambiente marinho, merecem destaque as algas pardas pertencentes à família Dictyotaceae e gênero Canistrocarpus. Este gênero foi instituído recentemente, e algumas algas anteriormente denominadas Dictyota foram reclassificadas como Canistrocarpus. O presente trabalho descreve os resultados do estudo ficoquímico de Canistrocarpus cervicornis (anteriormente classificada como Dictyota pulchella), envolvendo a avaliação do perfil ficoquímico de C. cervicornis coletada no litoral da Paraíba e no litoral do Rio de Janeiro. Este estudo contribuiu para a correta denominação da então D. pulchella em C. cervicornis. As amostras da alga foram coletadas na Praia do Bessa (coordenadas 07º04’01’’ S e 34º49’35’’ W) na cidade de João Pessoa (PB), em agosto de 2010 e em dezembro de 2012. A secagem foi realizada a temperatura ambiente, em local arejado e a extração procedida com CH2Cl2:MeOH (2:1). O extrato resultante da primeira coleta foi submetido a uma filtração a vácuo em sílica gel com os solventes hexano, acetato de etila e metanol em gradiente crescente de polaridade. A fração hexano:acetato de etila (85:15) foi submetida a cromatografia em coluna, de onde se obteve um novo produto natural, o diterpeno dolastano 4,7-diacetoxi de dichotenona A. O extrato resultante da segunda coleta foi dividido em duas partes. Uma alíquota foi submetida à cromatografia em coluna, de onde se obteve o diterpeno dolastano conhecido 4-acetoxi-14-hidroxidolasta-1(15),7,9-trieno. A segunda alíquota do extrato foi acetilada e submetida a cromatografias em coluna, o que levou ao isolamento de três diterpenos, sendo um dolastano inédito na literatura, o 4-desoxi-amijidictyol e dois secodolastanos conhecidos, o acetato de isolinearol e o indicol. Os constituintes químicos isolados foram identificados por análises espectroscópicas de RMN 1H e 13C uni e bidimensionais, rotação ótica, e comparação com dados da literatura. Este estudo contribuiu para a quimiotaxonomia da tribo Dictyoteae, em especial de C. cervicornis.

This thesis is the first study to provide a detailed characterization of Acropora cervicornis transplants and donor colony survival on southeast Florida coral reefs. Since May 2006 this species has been listed as a threatened species under the Endangered Species Act. As populations continue to decline restoration efforts need to be evaluated to determine if there is an effort that could facilitate a population rebound. The overall goal of this project was to examine potential Acropora cervicornis restoration techniques along the entire Florida reef tract including Broward County, Miami Dade County, and Monroe County. For my thesis I used a portion of the data collected from the Broward County region nursery. I analyzed data on the donor colonies and the nursery fragments. The goals were to determine if fragments generated from clippings removed from donor colonies can 1) be transplanted to a nursery site, 2) have acceptable survivorship and 3) increase in complexity (branching). Beyond survival and growth, I also examined genotypic differences in fragment survival and growth rates. Twelve A. cervicornis donor colonies separated by as much as 26 km were identified, and monitored quarterly for 19 months. From each donor colony three 10 cm clippings were removed for transplantation to the nursery habitat and one 1 cm clipping was taken for genetic analysis. Prior to transplantation, each 10 cm clipping was cut into 3 cm fragments. Transplantation occurred in September, October, and December 2007, transplanting 1/3 of the fragments horizontal and 2/3 vertical in orientation. Fragments in the nursery were monitored monthly through November 2008. During each monthly monitoring, images were taken, fragments were measured, branches were counted, and condition (partial mortality, disease, predation, etc.) was assessed. Each donor colony sampled with in Broward County for this project was determined to be a unique genotype using microsatellites. Significant differences in survival, growth, and number of branches were determined among fragment genotypes. Vertically orientated fragments had higher survivorship, but horizontal fragments had higher mean growth rates and number of branches per fragment. This coral restoration project has the real possibility of providing important information on the effectiveness of utilizing the asexual, fragmentation, capacity of A. cervicornis to facilitate A. cervicornis population conservation. My effort could contribute to a quantitative comparison of Acropora genotypic variation in survivorship and growth, which will provide information on intra- and inter-regional potential for large-scale restoration within the Florida reef tract.

This thesis provides a detailed analysis of the growth and survivorship of outplanted Acropora cervicornis corals from underwater nurseries within three regions of the Florida Reef Tract. Substantial loss of stony coral cover on Florida’s coral reefs, including the branching staghorn coral, Acropora cervicornis, has occurred for decades due to disturbances such as disease, temperature induced bleaching, hurricanes, sedimentation, and pollution. This rapid population decline contributed to A. cervicornis being listed as a threatened species under the U.S. Endangered Species Act in May 2006. To aid in the recovery of the species, coral fragments were grown in underwater nurseries and outplanted to selected sites located within unique cross-shelf zones in the Upper Florida Keys, Lower Keys, and Biscayne regions. This study evaluated the regional and zonal variation in growth and survivorship of known genotypes of outplanted A. cervicornis corals to better inform future large-scale restoration projects. The zone in which corals of A. cervicornis were outplanted to had a more significant effect on growth than the coral genotype. The forereef zone within the Upper and Lower Keys regions and the mid-channel zone in the Biscayne region had significantly higher mean growth rates. When comparing growth rates of genotypes that performed best, high growth, in the Lower Keys nursery, these same genotypes did not perform the best at any of the outplant sites. Survivorship was not significantly different in any of the regions. Based on these results, future coral outplantings focused in the forereef and mid-channel zones would maximize growth. Choosing coral genotypes based on their high growth rates in the nursery does not ensure the same high growth rates when outplanted.

This thesis is the first known study to relocate Acropora cervicornis across multiple regions of the Florida Reef Tract. Since 2006, A. cervicornis has been listed as a threatened coral species under the U.S. Endangered Species Act. In response, restoration efforts utilizing coral nursery methods have been implemented throughout the Caribbean. The primary objective of this research was to determine the response of A. cervicornis colonies to being relocated between two coral nurseries separated by approximately 150km along the Florida Reef Tract. To accomplish this, a reciprocal transport was conducted between coral fragments with known genotypes from Broward County and Monroe County, Florida. A subset of coral ramets (fragments of a single genotype) was removed from the nursery of origin and relocated to the opposing coral nursery, while the remaining ramets stayed in their original nursery to serve as controls. Following transplant, both relocated and non-relocated corals were monitored for 14 months and survivorship, growth rates, branching frequency, and coral condition data were collected. In addition, tissue samples were collected twice during the monitoring period to determine zooxanthellae densities. Reaction norms were used to predict the responses of each measured variable for each genotype in response to being relocated. Relocated coral colonies from both nurseries exhibited equal or greater survivorship than the non-relocated corals from their original nursery. Growth rates, branching frequency, and zooxanthellae densities were highest in the corals that were previously in or relocated to Broward County. Within each nursery, relocated and non-relocated corals were not significantly different in any of the measured parameters. Throughout the study period, there were no signs of disease, bleaching, or predation on any of the corals. These findings demonstrate that A. cervicornis colonies can be successfully relocated across regions of the Florida Reef Tract suggesting that colonies throughout the FRT may be used for collaborative restoration efforts. Reaction norm analysis indicated phenotypically plastic responses in each growth parameter with some instances of genotype-by-environment interactions. Finally, these results suggest the need for additional research to investigate regional differences in A. cervicornis populations for proper management and restoration approaches.

Rickettsiales-like organisms (RLOs) are thought to be related to bacteria in the order Rickettsiales. They have been reported to occur in the staghorn coral (Acropora cervicornis), and this study investigated trends of infection over time, and in relation to the health of infected corals. This study focuses on tissue samples taken mostly from visibly healthy A. cervicornis thickets in Broward County, Florida, and processed for histological examination. Samples were originally collected and analyzed to document reproduction during years 2001 through 2012, and tissue loss diseases (white-band disease [WBD] types I and II, and rapid tissue loss). The presence of suspect RLOs, the presence of ovoid bacterial aggregates in the basal body wall, and the condition of the coral tissue were examined in hematoxylin and eosin (H&E) and Giemsa-stained sections. Determination was made as to whether suspect RLO infection severity, location, or the presence of bacterial aggregates are correlated with changes in tissue condition associated with WBD. To better understand progression, trends, and periodicity in bacterial presence and coral tissue health, these data were then further analyzed for potential correlation with the month, year, and average monthly nighttime sea surface temperatures (categorized into ranges above, within, or below 24–29°C) when samples were collected. The severity of suspect RLO infections and the presence of bacterial aggregates in A. cervicornis varied over time, with no correlation with the location of suspect RLOs within the polyp. High suspect RLO prevalence was correlated with normal tissue conditions, while low suspect RLO prevalence was correlated with abnormal tissue conditions. However, high prevalence of bacterial aggregates was correlated to abnormal tissue conditions. Epidermal RLO and overall suspect RLO prevalence severity scores were significantly higher among samples collected when monthly average nighttime sea surface temperatures were below 24°C in contrast to samples collected when temperatures were between 24–29°C, suggesting direct or indirect effects of sea surface temperatures on infection severity. The areas of suspect RLO intracellular bodies within infected mucocytes were measured using digital image analysis software and found to be positively correlated with worsening coral tissue condition. Semiquantitative variable scores for histoslides stained with H&E were significantly different from those stained with Giemsa, indicating that these stains cannot be used interchangeably to study the presence of bacteria and the condition of coral tissue. Overall, the results of this study indicate that infection severity of suspect RLOs and the presence of bacterial aggregates are variable and correlated with the incidence of WBD-I in A. cervicornis. However, the exact nature of this relationship remains unclear. Further studies are necessary to interpret trends detected during this analysis to develop a better understanding of what contributes to the severe tissue-loss outbreaks and mortalities of A. cervicornis.