Bei Säugerkeimzellen, Zygoten und Embryos in frühen Stadien kommt der epigenetischen Neuprogammierung eine außergewöhnlich wichtige Rolle in der Regulation der Genomfunktionen in entscheidenden Entwicklungsstadien zu. Die epigenetische Neuprogrammierung in Keimzellen löscht zuerst die Imprinting-Markierungen und Epi-Mutationen und stellt dann geschlechtsspezifische Markierungen (genomische Prägung) wieder her. Die vorliegende Arbeit bezieht sich auf das Löschen epigenetischer Modifikationen in primordialen Mauskeimzellen (primordial germ cells (PGCs)) zwischen dem 10.5 bis 13.5 Tag nach der Befruchtung. Entgegen früheren Annahmen zeigen unsere Ergebnisse, daß primordiale Mauskeimzellen (PGCs) beim Eintritt in die embryonalen Keimdrüsen noch immer DNS Methylierungsmarker besitzen, die ähnlich dem Marker in somatischen Zellen sind. Kurz nach dem Eintritt in die Keimdrüsen werden die DNS Methylierungsmarker, die in Verbindung mit geprägten und nicht geprägten Genen stehen, gelöscht. Für die Mehrzahl der Gene beginnt die Löschung der Marker in männlichen und weiblichen Embryos gleichzeitig und ist innerhalb eines Entwicklungstages abgeschlossen. Diese Kinetik deutet auf einen aktiven Demethylierungsprozess hin, initiiert durch ein somatisches Signal, ausgehend von der embryonalen Keimdrüse. Der Zeitpunkt der Neuprogrammierung in den primordialen Keimzellen ist entscheidend, da er sicherstellt, daß Keimzellen beiden Geschlechts einen epigenetisch äquivalenten Status erhalten, bevor sie geschlechtsspezifisch ausdifferenzieren und anschließend neu elterlich geprägt werden. Vollständiges Verständnis des Prozesses der Neuprogrammierung der Keimzellen ist nicht nur im Hinblick auf genomisches Imprinting wichtig, sondern auch für die Erforschung von Mechanismen für die Wiederherstellung von omnipotenten Zellen bei Klonierung und Stammzellenerhaltung. / Epigenetic reprogramming in mammalian germ cells, zygote and early embryos, plays a crucial role in regulating genome functions at critical stages of development. Germ line epigenetic reprogramming assures erasure of all the imprinting marks and epi-mutations and establishment of new sex-specific gametic imprints. The presented work focuses on the erasure of epigenetic modifications that occur in mouse primordial germ cells (PGCs) between day 10.5 to 13.5 post coitum (dpc). Contrary to previous assumptions, our results show that as they enter the genital ridge the PGCs still possess DNA methylation marks comparable to those found in somatic cells. Shortly after the entry of PGCs into the gonadal anlagen the DNA methylation marks associated with imprinted and non-imprinted genes are erased. For most genes the erasure commences simultaneously in PGCs of both male and female embryos and is completed within only one day of development. The kinetics of this process indicates that is an active demethylation process initiated by a somatic signal emanating from the stroma of the genital ridge. The timing of reprogramming in PGCs is crucial since it ensures that germ cells of both sexes acquire an equivalent epigenetic state prior to the differentiation of the definitive male and female germ cells in which, new parental imprints are established subsequently. Complete understanding of the germline reprogramming processes is important not only in the light of genomic imprinting but also for resolving other mechanisms connected with restoring cellular totipotency, such as cloning and stem cell derivation.

Uma das espécies de coral pétreo zooxantelado com mais ampla distribuição no litoral brasileiro é Madracis decactis Lyman, 1859. M. decactis forma colônias incrustantes nodulares que podem atingir até 30 cm de diâmetro. O estudo da gametogênese foi realizado através de coletas bimensais na Baía de Ilha Grande, RJ durante 21 meses (agosto/2004-maio/2006), totalizando 10 coletas (12 colônias/coleta) (Licença IBAMA no. 201/2004). Foram realizados cortes histológicos de 7 µm, e de 10 até 16 pólipos por colônia foram analisados, totalizando mais de 1800 pólipos. A análise dos pólipos indicou que M. decactis é hermafrodita, com gametas localizados no mesmo lóculo gástrico, porém em mesentérios diferentes. A gametogênese durou cerca de sete meses. A ovogênese iniciou-se ao redor de outubro, enquanto que a espermatogênese teve inicio no final de fevereiro, ambas terminando em sincronia no final do mês de maio. O exame dos pólipos férteis indicou a presença dos estágios I, II e III de desenvolvimento para a ovogênese e dos estágios I, II, III e IV para a espermatogênese. Não foram encontrados embriões ou plânulas nos cortes histológicos, indicando talvez que estes estágios permaneçam pouco tempo no interior do pólipo. O pico da atividade reprodutiva ocorre entre os meses de fevereiro e abril com todas as colônias férteis contendo ovócitos principalmente no estágio III de maturação. A provável época de liberação de plânulas ocorre entre os meses de abril e maio, sob influencia das condições ambientais como a temperatura da água do mar, a irradiação solar e a pluviosidade. O presente trabalho forneceu informações básicas a respeito da biologia reprodutiva de Madracis decactis presente em Ilha Grande, sendo uma contribuição para outros estudos mais específicos na área de manejo e conservação de ambientes marinhos. / One of the most wide distributed coral species along the Brazilian cost is Madracis decactis Lyman, 1859. M. decactis growths as nodular incrusting colonies that can reach up to 30 cm in diameter. In order to study the gametogenesis cycle, bimonthly collections were done at Ilha Grande Bay, Rio de Janeiro, during 21 months (August/2004- May/2006), totaling 10 collections (12 colonies/collection) (License no. 201/2004). Histological sections of 7 µm thick were done to analyze 10 to 16 polyps per colony (more than 1800 polyps in total). The analysis indicated that M. decactis is hermaphroditic, with male and female gametes developing at the same gastric loculi, but in different mesenteries. The gametogenesis lasted about 7 months, the oogenesis starts at October , while spermtogenesis starts at the end of February, both reaching the maturity in synchrony at the end of May. The exam of fertile polyps indicated the presence of stages I, II and III for oogenesis and I, II, III and IV for spermatogenesis. No embryo or planula were observed in the histological sections, indicating that maybe these stages stay for a short period inside the polyp cavity. The peak of reproductive activity was between February and April when all the polyps were fertile containing manly stage III oocytes. The releasing of planulae may happen between April and May, under the influence of environmental conditions as sea water temperature, solar insolation and rainy season. The present study has provided basic information about the reproductive biology of Madracis decactis from Ilha Grande Bay, and it may be a contribution to further studies about management and conservation of marine environments.

Nas últimas décadas tem se dado maior atenção à reprodução de animais silvestres, visto que para se conhecer e preservar uma espécie se faz necessário o conhecimento da história natural aliado aos processos da fisiologia e morfologia reprodutivas. Ciclos reprodutivos podem ser sazonais ou assazonais e apresentar variações em ambos os padrões. Por sua natureza e diversidade de fatores associados (e.g., atividade, dieta, temperatura), as estratégias reprodutivas vêm sendo investigadas sob uma abordagem evolutiva. Estudos do comportamento reprodutivo e dimorfismo sexual também têm sido destacados nesse âmbito. No entanto, poucos trabalhos são realizados para se conhecer as estratégias reprodutivas em serpentes neotropicais. Na maioria das vezes tais trabalhos abordam apenas alguns aspectos do ciclo reprodutivo de serpentes, como época de nascimentos, cópula e gestação, podendo ainda ser descritos de forma divergente pelo fato de muitos destes estudos serem norteados por dados de serpentes em cativeiro. Estudos microscópicos e macroscópicos, delineados com animais recém-chegados da natureza e preservados em coleções científicas, pode fornecer uma descrição e uma análise mais efetiva do ciclo reprodutivo permitindo a integração destes fatores. Assim, este trabalho investigou diversos aspectos da biologia reprodutiva de fêmeas e machos de populações de Bothrops alternatus do Brasil. Além disso, foram analisadas e descritas as estratégias reprodutivas empregadas por ambos os sexos durante o ciclo reprodutivo anual. Para isso, os espécimes foram dissecados ventralmente retirando-se fragmentos das gônadas e vias genitais de fêmeas e machos, em diferentes épocas do ano, a fim de se identificar as principais fases do ciclo vitelogênico, espermatogênico e do segmento sexual renal. Igualmente, para determinar se há estocagem de esperma nas fêmeas e nos machos, será investigada o útero aglandular e região do infundíbulo, vias genitais e ductos deferentes respectivamente. Tais dados permitirão compor um padrão reprodutivo, para essa espécie, levando-se em conta processos morfofisiológicos em ambos os sexos. Também foi realizada uma comparação dos ciclos reprodutivos entre populações de Bothrops alternatus e análises de possíveis influências ambientais na duração dos eventos reprodutivos / Over the last decades a greater attention has been given to the reproduction of wild animals, as the knowledge of natural history combined with the processes of reproductive physiology and morphology is essential for the preservation of a species. Reproductive cycles may be seasonal or not, and both patterns may have variations. Due to its nature and to the diversity of the associated factors (e.g., activity, diet, temperature), reproductive strategies have been investigated in an evolutionary approach. Studies of reproductive behavior and sexual dimorphism have also been highlighted in this context. However, few studies have been conducted to determine reproductive strategies in neotropical snakes. Most studies address only some aspects of the reproductive cycle of snakes, such as time of birth, copulation and pregnancy, and may present divergent descriptions as many of them are based on data from snakes in captivity. Microscopic and macroscopic studies carried out with animals newcomers from nature and preserved in scientific collections can provide a more effective analysis of the reproductive cycle allowing the integration of these factors. In this context, this project aims to investigate various aspects of the reproductive biology of female and male populations of Bothrops alternatus from Brazil. The reproductive strategies employed by both sexes during the annual reproductive cycle was analyzed and described. The specimens was dissected ventrally and fragments of gonads and genital tracts of females and males was removed at different times of the year in order to identify the main phases of the vitellogenic cycle, spermatogenesis and sexual segment of the kidney. Also, to determine whether sperm storage is done by females and males, we will investigate the aglandular uterus, the posterior infundibulum, genital tracts and vas deferens respectively. Such data was compose a reproductive pattern for the species that will take into account the morphophysiologic processes of both sexes. It was also performed a comparison of reproductive cycles between populations of Bothrops alternates and analysis of possible environmental influences in the duration of reproductive events

Hui, Yuk Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 132-146). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract (English) --- p.ii / Abstract (Chinese) --- p.vi / Contents --- p.viii / List of Tables --- p.xi / List of Figures --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- General biology of scleractinian corals --- p.1 / Chapter 1.2 --- Global coral degradation --- p.2 / Chapter 1.3 --- Effects of temperature and other environmental factors on coral growth and development --- p.4 / Chapter 1.4 --- Restoration strategies --- p.6 / Chapter 1.5 --- Scleractinian corals in Hong Kong and study site of this thesis research --- p.9 / Chapter 1.6 --- Target coral species --- p.10 / Chapter 1.7 --- Objectives --- p.11 / Chapter 1.8 --- Thesis outline --- p.12 / Chapter Chapter 2 --- "Gametogenic Cycles of Acropora tumida in Cheung Sha Wan,Tung Ping Chau, HKSAR" --- p.17 / Chapter 2.1 --- Introduction --- p.17 / Chapter 2.2 --- Methods and Materials --- p.20 / Chapter 2.2.1 --- Samples collection --- p.20 / Chapter 2.2.2 --- Histology --- p.20 / Chapter 2.2.3 --- Gamete classification and measurements --- p.21 / Chapter 2.2.4 --- Temperature records --- p.23 / Chapter 2.2.5 --- Data analysis --- p.23 / Chapter 2.3 --- Results --- p.23 / Chapter 2.3.1 --- Gametogenic cycle of Acropora tumida --- p.23 / Chapter 2.3.2 --- Oogenesis in Acropora tumida --- p.24 / Chapter 2.3.3 --- Spermatogenesis in Acropora tumida --- p.29 / Chapter 2.3.4 --- Temperature data --- p.30 / Chapter 2.4 --- Discussion --- p.31 / Chapter 2.4.1 --- Gametogenic cycle of Acropora tumida --- p.31 / Chapter 2.4.2 --- Oogenesis in Acropora tumida --- p.32 / Chapter 2.4.3 --- Spermatogenesis in Acropora tumida --- p.34 / Chapter Chapter 3 --- Comparison on the Oogenesis in Acropora tumida Grown Ex situ and In situ --- p.44 / Chapter 3.1 --- Introduction --- p.44 / Chapter 3.2 --- Methods and Materials --- p.48 / Chapter 3.2.1 --- Samples collection --- p.48 / Chapter 3.2.2 --- Histological analysis --- p.50 / Chapter 3.2.3 --- Environmental conditions --- p.50 / Chapter 3.2.4 --- Data analysis --- p.51 / Chapter 3.3 --- Results --- p.52 / Chapter 3.3.1 --- Comparison of oogenesis in ex situ and in situ Acropora tumida samples --- p.52 / Chapter 3.3.2 --- Comparison of spermary development in ex situ and in situ Acropora tumida samples --- p.54 / Chapter 3.3.3 --- Environmental conditions --- p.54 / Chapter 3.4 --- Discussion --- p.55 / Chapter Chapter 4 --- Oogenesis along a Fragmentation Gradient in the Branching coral Acropora tumida --- p.70 / Chapter 4.1 --- Introduction --- p.70 / Chapter 4.2 --- Methods and Materials --- p.74 / Chapter 4.2.1 --- Samples collection and histological analysis --- p.74 / Chapter 4.2.2 --- Data analysis --- p.76 / Chapter 4.3 --- Results --- p.76 / Chapter 4.3.1 --- Oogenesis in intact Acropora tumida colonies --- p.76 / Chapter 4.3.2 --- Oogenesis in Acropora tumida fragments --- p.77 / Chapter 4.3.3 --- Sterile zone in Acropora tumida branch tips --- p.78 / Chapter 4.4 --- Discussion --- p.79 / Chapter Chapter 5 --- "Temperature Effects on the Survivorship, Growth and Oogenesis in Acropora tumida Fragments Grown under Laboratory Conditions " --- p.87 / Chapter 5.1 --- Introduction --- p.87 / Chapter 5.2 --- Methods and Materials --- p.91 / Chapter 5.2.1 --- Samples collection --- p.91 / Chapter 5.2.2 --- Samples culture --- p.92 / Chapter 5.2.3 --- Survivorship --- p.93 / Chapter 5.2.4 --- Growth measurement --- p.94 / Chapter 5.2.5 --- Reproductive analysis --- p.94 / Chapter 5.2.6 --- Statistical analysis --- p.95 / Chapter 5.3 --- Results --- p.95 / Chapter 5.3.1 --- Temperatures in ex situ ambient aquaria and the in situ site at Tung Ping Chau --- p.95 / Chapter 5.3.2 --- Survivorship of coral branches --- p.96 / Chapter 5.3.3 --- Growth measurement --- p.97 / Chapter 5.3.4 --- Oogenic development --- p.98 / Chapter 5.3.4.1 --- Oocyte developmental stages --- p.99 / Chapter 5.3.4.2 --- Oocyte size measurement --- p.100 / Chapter 5.3.4.3 --- Oocyte density --- p.103 / Chapter 5.3.5 --- Spermary developmental stages --- p.105 / Chapter 5.4 --- Discussion --- p.105 / Chapter 5.4.1 --- Survivorship --- p.105 / Chapter 5.4.2 --- Growth measurement --- p.107 / Chapter 5.4.3 --- Oogenesis under different temperature conditions --- p.109 / Chapter 5.4.3.1 --- Oocyte developmental stage --- p.109 / Chapter 5.4.3.2 --- Oocyte size measurement and density --- p.113 / Chapter 5.4.5 --- Spermary developmental stage --- p.114 / Chapter Chapter 6 --- Summary and Perspectives --- p.128 / References --- p.132

This study examined patterns of coral reproduction and recruitment on lagoonal reefs adjacent to Mombasa in Kenya, at latitude 4ºS. Very little detailed research has been done on the reproductive patterns of scleractinian corals on equatorial reefs, where it has been suggested that seasonality and spawning synchrony may break down due to the weak environmental cues that are thought to govern the onset and timing of reproduction. Gametogenic data were collected for three faviid (Echinopora gemmacea, Platygyra daedalea and Leptoria phrygia) and three Acropora species (A. tenuis, A. valida and Acropora sp.1) in the Mombasa Marine National Park and Reserve between April 2003 – May 2005. A further 20 species of Acropora were identified (9 species represented range extensions) and marked to examine intra- and inter-specific spawning synchrony within this genus. In comparison to other regions, the overall pattern of coral reproduction in Kenya was found to be asynchronous, with spawning occurring over 9 months of the year from August – April, with some level of ‘temporal reproductive isolation’ occurring between species in relation to the main lunar month and lunar quarter when spawning occurred. Proximate cues governing the timing of reproduction could not be clearly discerned in Kenya with spawning occurring during both rising and maximum temperatures, during both neap and spring tides and across all lunar phases. Acropora species spawned over a 7-month period between October – April and faviid species over a 5-month period from December - April. The timing of reproduction in Acropora varied both within and among species, with the main release of gametes occurring from January – March when sea surface temperatures were at their summer maximum. Individual species released gametes over 2-5 months. The greatest overlap in spawning Acropora species occurred in February, which coincided with the spawning months of P. daedalea and E. gemmacea and suggests that some degree of multispecific spawning is a characteristic of Kenyan reefs. Within the main spawning period individual Acropora species had their main spawning in different lunar months. Acropora species released gametes in all lunar quarters, with the highest number of colonies and species spawning in the 3rd lunar quarter (i.e. in the 7 nights after full moon). Spawning in the faviids was more synchronised than Acropora species with the majority of faviid corals spawning in the 3rd lunar quarter. Single annual cycles of gametogenesis were recorded in E. gemmacea, A. tenuis, L.phrygia, most colonies of A. valida and Acropora sp.1, and in 84% of P. daedalea colonies. Biannual cycles of gametogenesis were recorded in 16% of P.daedalea colonies, which included two morphotypes identified in the Mombasa lagoon through morphometric and genetic studies. The presence of different oocyte sizes in L.phrygia during gametogenesis suggested that in some colonies there were two slightly overlapping oogenic cycles, which terminated in spawning within 1-2 months of each other. Overlapping oogenic cycles have not previously been recorded in hermaphroditic broadcast spawning corals in the tropics. The findings from Kenya support the hypothesis of protracted breeding seasons and a breakdown of spawning synchrony nearer the equator. It is hypothesised that the high fecundities recorded in faviid and Acropora species in Kenya compared to other regions, may allow reef corals to stagger their reproduction over 2-5 months, without incurring a significant reduction in fertilisation rates. Spat from the Family Pocilloporidae dominated settlement tiles in the Marine National Park and Reserve comprising 93.7% of spat, which contrasts with other tropical reefs where Acroporidae spat dominate. Patterns of settlement of Acroporidae spat generally coincided with the timing and extended spawning season in Acropora species in Kenya. The density and relative composition of coral recruits and juvenile corals on natural substrata recorded during this study were similar to those recorded before the 1997-98 bleaching event. There is no evidence to suggest that Kenya’s reefs have undergone a phase-shift in community structure, and reef recovery is occurring post-bleaching with mean percent hard coral cover currently at 25%. The slow rate of recovery of Kenya’s reefs is likely to reflect the scale of the mortality, source and availability of coral larvae as well as post-settlement processes operating at individual sites. In the medium-term, the recovery of Kenya’s reefs appears to be more strongly dependent on larvae from local reefs.

This study examined patterns of coral reproduction and recruitment on lagoonal reefs adjacent to Mombasa in Kenya, at latitude 4ºS. Very little detailed research has been done on the reproductive patterns of scleractinian corals on equatorial reefs, where it has been suggested that seasonality and spawning synchrony may break down due to the weak environmental cues that are thought to govern the onset and timing of reproduction. Gametogenic data were collected for three faviid (Echinopora gemmacea, Platygyra daedalea and Leptoria phrygia) and three Acropora species (A. tenuis, A. valida and Acropora sp.1) in the Mombasa Marine National Park and Reserve between April 2003 – May 2005. A further 20 species of Acropora were identified (9 species represented range extensions) and marked to examine intra- and inter-specific spawning synchrony within this genus. In comparison to other regions, the overall pattern of coral reproduction in Kenya was found to be asynchronous, with spawning occurring over 9 months of the year from August – April, with some level of ‘temporal reproductive isolation’ occurring between species in relation to the main lunar month and lunar quarter when spawning occurred. Proximate cues governing the timing of reproduction could not be clearly discerned in Kenya with spawning occurring during both rising and maximum temperatures, during both neap and spring tides and across all lunar phases. Acropora species spawned over a 7-month period between October – April and faviid species over a 5-month period from December - April. The timing of reproduction in Acropora varied both within and among species, with the main release of gametes occurring from January – March when sea surface temperatures were at their summer maximum. Individual species released gametes over 2-5 months. The greatest overlap in spawning Acropora species occurred in February, which coincided with the spawning months of P. daedalea and E. gemmacea and suggests that some degree of multispecific spawning is a characteristic of Kenyan reefs. Within the main spawning period individual Acropora species had their main spawning in different lunar months. Acropora species released gametes in all lunar quarters, with the highest number of colonies and species spawning in the 3rd lunar quarter (i.e. in the 7 nights after full moon). Spawning in the faviids was more synchronised than Acropora species with the majority of faviid corals spawning in the 3rd lunar quarter. Single annual cycles of gametogenesis were recorded in E. gemmacea, A. tenuis, L.phrygia, most colonies of A. valida and Acropora sp.1, and in 84% of P. daedalea colonies. Biannual cycles of gametogenesis were recorded in 16% of P.daedalea colonies, which included two morphotypes identified in the Mombasa lagoon through morphometric and genetic studies. The presence of different oocyte sizes in L.phrygia during gametogenesis suggested that in some colonies there were two slightly overlapping oogenic cycles, which terminated in spawning within 1-2 months of each other. Overlapping oogenic cycles have not previously been recorded in hermaphroditic broadcast spawning corals in the tropics. The findings from Kenya support the hypothesis of protracted breeding seasons and a breakdown of spawning synchrony nearer the equator. It is hypothesised that the high fecundities recorded in faviid and Acropora species in Kenya compared to other regions, may allow reef corals to stagger their reproduction over 2-5 months, without incurring a significant reduction in fertilisation rates. Spat from the Family Pocilloporidae dominated settlement tiles in the Marine National Park and Reserve comprising 93.7% of spat, which contrasts with other tropical reefs where Acroporidae spat dominate. Patterns of settlement of Acroporidae spat generally coincided with the timing and extended spawning season in Acropora species in Kenya. The density and relative composition of coral recruits and juvenile corals on natural substrata recorded during this study were similar to those recorded before the 1997-98 bleaching event. There is no evidence to suggest that Kenya’s reefs have undergone a phase-shift in community structure, and reef recovery is occurring post-bleaching with mean percent hard coral cover currently at 25%. The slow rate of recovery of Kenya’s reefs is likely to reflect the scale of the mortality, source and availability of coral larvae as well as post-settlement processes operating at individual sites. In the medium-term, the recovery of Kenya’s reefs appears to be more strongly dependent on larvae from local reefs.

This study examined patterns of coral reproduction and recruitment on lagoonal reefs adjacent to Mombasa in Kenya, at latitude 4ºS. Very little detailed research has been done on the reproductive patterns of scleractinian corals on equatorial reefs, where it has been suggested that seasonality and spawning synchrony may break down due to the weak environmental cues that are thought to govern the onset and timing of reproduction. Gametogenic data were collected for three faviid (Echinopora gemmacea, Platygyra daedalea and Leptoria phrygia) and three Acropora species (A. tenuis, A. valida and Acropora sp.1) in the Mombasa Marine National Park and Reserve between April 2003 – May 2005. A further 20 species of Acropora were identified (9 species represented range extensions) and marked to examine intra- and inter-specific spawning synchrony within this genus. In comparison to other regions, the overall pattern of coral reproduction in Kenya was found to be asynchronous, with spawning occurring over 9 months of the year from August – April, with some level of ‘temporal reproductive isolation’ occurring between species in relation to the main lunar month and lunar quarter when spawning occurred. Proximate cues governing the timing of reproduction could not be clearly discerned in Kenya with spawning occurring during both rising and maximum temperatures, during both neap and spring tides and across all lunar phases. Acropora species spawned over a 7-month period between October – April and faviid species over a 5-month period from December - April. The timing of reproduction in Acropora varied both within and among species, with the main release of gametes occurring from January – March when sea surface temperatures were at their summer maximum. Individual species released gametes over 2-5 months. The greatest overlap in spawning Acropora species occurred in February, which coincided with the spawning months of P. daedalea and E. gemmacea and suggests that some degree of multispecific spawning is a characteristic of Kenyan reefs. Within the main spawning period individual Acropora species had their main spawning in different lunar months. Acropora species released gametes in all lunar quarters, with the highest number of colonies and species spawning in the 3rd lunar quarter (i.e. in the 7 nights after full moon). Spawning in the faviids was more synchronised than Acropora species with the majority of faviid corals spawning in the 3rd lunar quarter. Single annual cycles of gametogenesis were recorded in E. gemmacea, A. tenuis, L.phrygia, most colonies of A. valida and Acropora sp.1, and in 84% of P. daedalea colonies. Biannual cycles of gametogenesis were recorded in 16% of P.daedalea colonies, which included two morphotypes identified in the Mombasa lagoon through morphometric and genetic studies. The presence of different oocyte sizes in L.phrygia during gametogenesis suggested that in some colonies there were two slightly overlapping oogenic cycles, which terminated in spawning within 1-2 months of each other. Overlapping oogenic cycles have not previously been recorded in hermaphroditic broadcast spawning corals in the tropics. The findings from Kenya support the hypothesis of protracted breeding seasons and a breakdown of spawning synchrony nearer the equator. It is hypothesised that the high fecundities recorded in faviid and Acropora species in Kenya compared to other regions, may allow reef corals to stagger their reproduction over 2-5 months, without incurring a significant reduction in fertilisation rates. Spat from the Family Pocilloporidae dominated settlement tiles in the Marine National Park and Reserve comprising 93.7% of spat, which contrasts with other tropical reefs where Acroporidae spat dominate. Patterns of settlement of Acroporidae spat generally coincided with the timing and extended spawning season in Acropora species in Kenya. The density and relative composition of coral recruits and juvenile corals on natural substrata recorded during this study were similar to those recorded before the 1997-98 bleaching event. There is no evidence to suggest that Kenya’s reefs have undergone a phase-shift in community structure, and reef recovery is occurring post-bleaching with mean percent hard coral cover currently at 25%. The slow rate of recovery of Kenya’s reefs is likely to reflect the scale of the mortality, source and availability of coral larvae as well as post-settlement processes operating at individual sites. In the medium-term, the recovery of Kenya’s reefs appears to be more strongly dependent on larvae from local reefs.

Infectious diseases continue to be a major concern worldwide. They are the second leading cause of death after heart disease. Factors such as an increasing global population, travel, urbanization, global climate change and evolution of pathogens have made infectious diseases more common. Infectious diseases, particularly neglected tropical diseases (NTDs) result in many deaths worldwide. Malaria and leishmaniasis are two common (NTDs) which affect low income countries around the globe. Low cost drugs with novel mechanism of action are required to tackle the growing resistances of parasites against current drugs used in the developing world, where most of the cases occur. The first part of this manuscript (chapters 1 - 3) describes the synthesis of novel analogs active against Leishmania donovani parasite which causes leishmaniasis. Leishmaniasis is a vector-borne complex group of diseases transmitted through the bite of an infected female sand-fly. Its clinical manifestations range from the less severe (cutaneous) to fatal (visceral) forms depending upon infecting species, immunity of host and the environment. Reports have suggested the role of Heat shock protein 90 (Hsp 90) in the differentiation of the Leishmania parasite from the promastigote stage to the pathogenic amastigote stage inside the host. A series of tetrahydro-indazole, tetrahydro-pyrazolo pyridine and radicicol hybrid compounds were prepared based on known Hsp 90 inhibitors, SNX2112 and NVP-AUY922. The synthetic approach allowed us to generate a diverse library of analogs which were used to probe the hydrophobic pocket of Hsp 90 active site. The most active compound, was found to be twice more active as the clinically used drug, Miltefosine, in an infected macrophage assay with an IC50 = 0.88 µM. The second part of this manuscript (chapters 4 - 5) describes the synthesis of xanthurenic acid analogs as antimalarial drugs. Xanthurenic acid (XA) is a vital component for the gametogenesis of the Plasmodium inside the mosquito’s gut. Gametogenesis plays an important part in the continuation of the parasite’s life cycle. A series of xanthurenic acid analogs were synthesized with the aim of inducing premature exflagellation of the microgametes, thus blocking the key step required for the transmission of parasites from humans to the mosquito. A biotinylated xanthurenic acid analog and a clickable xanthurenic acid analog were also synthesized which will help us investigate the mechanism of action of xanthurenic acid in inducing gametogenesis in mosquito. In the preliminary screening efforts in an exflagellation assay, analog 4.40 showed promising activity and was more active in inducing exflagellation than xanthurenic acid. An exflagellation assay of other analogs is currently being pursued. Further investigations into the molecular target and mechanism of action are underway with the biotinylated xanthurenic acid analog.

From January 2012 to December 2012 I collected adult oysters from two intertidal populations on a monthly basis in the Coos Bay estuary, Oregon for histological analysis of their gonads. Gametogenesis and spawning occur seasonally from May through September, when water temperatures exceed 14.5&deg C, with brooding oysters found from July through September. Oocyte diameters increased significantly from May to June, and from June to July within oyster populations at Haynes Inlet and Coalbank Slough, respectively. Male gametogenesis initiated in May at Haynes Inlet and in June at Coalbank Slough. Dry meat condition values increased significantly during periods of reproduction and decreased following the reproductive season's end. Condition index values for Coalbank Slough were consistently lower than those at Haynes Inlet, suggesting poor nutrition or physiological stress. Salinities below recorded physiological thresholds are believed to be the primary environmental factor influencing the discrepancy in reproductive activity at Coalbank Slough.

O presente trabalho analisou as etapas de formação de rudimentos seminais e grãos de pólen de Passiflora elegans e P. haematostigma. As espécies foram analisadas fazendo-se o uso de microscopia fotônica e microscopia eletrônica de varredura e transmissão. O gineceu apresenta um estigma com emergências estigmáticas, estilete sólido e ovário tricarpelar e unilocular. O ovário apresenta inúmeros rudimentos seminais que formam-se a partir de divisões periclinais da camada central. Os rudimentos são crassinucelados e os tegumentos têm origem dérmica. A célula arquesporial divide-se mitoticamente antes da meiose. A meiose resulta em uma tétrade linear, com ginósporo calazal viável. A ginogametogênese é do tipo Poligonum e, durante a diferenciação do ginófito, os estratos parietais do nucelo são parcialmente consumidos e a epiderme nucelar divide-se periclinalmente na porção apical. As antípodas são efêmeras e o ginófito maduro apresenta duas sinérgides com aparelho fibrilar desenvolvido, oosfera e núcleo secundário no pólo micropilar. O rudimento seminal é anátropo, arilado, bitegumentado e com micrópila em “zig-zag”. O androceu é constituído por cinco anteras tetrasporangiadas. Os estratos parietais desenvolvem-se do tipo Dicotiledôneo e as camadas médias são persistentes na antera madura de P. haematostigma. As células do tapete são poliplóides e colapsam liberando os conteúdos no interior do lóculo. A citocinese é do tipo simultânea e as tétrades são tetraédricas na maioria. O pólen é liberado na fase bicelular, apresenta número de colpos variável em P. elegans e seis colporos em P. haematostigma. A esporoderme não apresenta camada basal em P. elegans e é mais espessa do que em P. haematostigma. Plastídios e mitocôndrias são englobados durante a formação da célula generativa e persistem até a fase de pólen maduro. / The present work analyzed the different developmental stages of the ovule and of the pollen grain in Passiflora elegans and P. haematostigma. The species were analyzed using photonic microscopy and scanning and transmission electron microscopy. The gynoecium has a stigma with stigmatic outgrowths, solid style and tricarpelar and unilocular ovary. The ovary has several seminal rudiments that are originated from the periclinal divisions of the central layer. The ovule is crassinucelate and the dermal layer originates the integuments. The arquesporial cell divides mitotically before meiosis. The meiosis results into a linear tetrad and only the chalazal gynospore is viable. The gynogametogenesis is of the Poligonum type, the nucellus parietal layers are partially degraded during the gynophyte differentiation, and nuclear epidermis divides periclinally in the apical portion. The antipodal cells are ephemeral and the mature gynophyte has two synergids with a developed filiform apparatus, egg cell and secondary nuclei in the micropilar pole. The ovule is anatropous, arillate, bitegmic and with zig-zag micropile. The androecium is constituted of five tetrasporangiate anthers. The parietal layers develop like the Dicotiledoneous type and middle layers are persistent in the mature anther of P. haematostigma. The tapetal cells are polyploids and collapse releasing their contents to the locus. The cytokinesis is of the simultaneous type and most of the tetrads are tetrahedral. The pollen is released during the bicellular stage, has a variable number of colpus in P. elegans and six colporus in P. haematostigma. The sporoderm does not have a basal layer in P. elegans and is thicker than in P. haematostigma. Plastids and mitochondria are enclosed during the generative cell formation and persist until the mature pollen stage.