A acidificação oceânica, resultante do aumento da concentração de CO2 atmosférico, vem alterando a química dos oceanos resultando na diminuição de seu pH. Diversos estudos avaliaram as consequências dessa diminuição no pH oceânico nas taxas de calcificação, reprodução e desenvolvimento em diversos modelos marinhos, entretanto estudos relacionados a outros processos fisiológicos, como a imunidade, e estudos com indivíduos adultos são escassos. Ouriços-do-mar são espécies aderidas ao substrato, importantes para a ciclagem de nutrientes no ambiente marinho, sendo também animais utilizados como bioindicadores para monitoramento ambiental; assim o estudo da resposta imune inata desses animais frente à acidificação dos oceanos é de extrema importância para prever possíveis alterações fisiológicas desses animais e sua capacidade de adaptação. O presente trabalho teve como objetivo avaliar as alterações provocadas pela acidificação oceânica na resposta imune e no balanço ácido base de duas espécies de ouriço-do-mar tropicais: Lytechinus variegatus e Echinometra lucunter durante as estações de verão e inverno; para isso foram analisados os índices fagocíticos, a capacidade de adesão e espraiamento celular dos amebócitos fagociticos além do balanço acido base do liquido celomático após o período de 24 horas e cinco dias de exposição aos pHs 7,6 e 7,3. Foi também avaliada a capacidade de recuperação dessas espécies com o objetivo de verificar se os parâmetros alterados pela exposição conseguiam ser reestabelecidos. Os resultados mostraram que a redução no pH da água do mar alterou a proporção celular, reduziu a capacidade de fagocitose e espraiamento dos amebócitos fagocíticos assim como também afetou o balanço ácido-base do líquido celomático. Foram encontradas diferenças também entre as estações do ano sendo estas encontradas apenas na espécie Lytechinus variegatus. O teste de recuperação mostrou que os parâmetros alterados pela exposição tendem a retornar aos valores controles, mostrando que em curto prazo essas alterações podem não ser irreversíveis, entretanto, mais estudos são necessários principalmente avaliando períodos de exposição prolongados. Juntos nossos resultados mostram que a acidificação oceânica prejudica parâmetros imunes extremamente importantes para a eliminação de patógenos e consequentemente a sobrevivência desses animais em um futuro oceano acidificado. / Ocean acidification due to increased atmospheric CO2 concentration is altering ocean chemistry resulting in the decrease of its pH. Several studies evaluated the effects of this decrease in ocean pH on calcification rates, reproduction and development in different marine models, however studies related to other physiological processes such as immunity and studies with adult animals are scarce. Sea urchins are species adhered to the substrate, important for nutrient cycling in the marine environment, also being used as bioindicators for environmental monitoring. Thus the study of the innate immune response of these animals due to the acidification of the oceans is extremely important to predict possible physiological changes of these animals and their ability to adapt to this condition. This study aimed to evaluate the changes caused by ocean acidification in the immune response and in the acid-base balance of two sea urchin tropical species: Lytechinus variegatus and Echinometra lucunter during seasons of summer and winter; for this, indexes of phagocytosis, cell adhesion and spreading ability were analyzed in addition to the acid-base balance of the coelomic fluid after 24 hours and five days of exposure to pH 7.6 and 7.3The recover ability of these species were also evaluated in order to verify if the parameters altered by exposition could be reestablished. The results shows that a reduction in the seawater pH changed the cell proportion, reduced the ability of phagocytosis and phagocytic amoebocyte spreading as well as affected the acid-base balance of the coelomic fluid. Differences were also found between seasons, but only in the specie Lytechinus variegatus. The recovery test showed that the parameters altered by exposure tend to return to control values, showing that in the short term these changes may not be irreversible, however, further studies are necessary mainly those related to prolonged periods of exposure. Together our results show that ocean acidification impairs immune parameters extremely important for the elimination of pathogens and consequently the survival of these animals in a future acidified ocean.

Submitted by Viviane Lima da Cunha (viviane@biblioteca.ufpb.br) on 2016-03-30T12:47:08Z No. of bitstreams: 1 arquivototal.pdf: 5765501 bytes, checksum: d810d02b334ee8a11679326d3a4f57aa (MD5) / Made available in DSpace on 2016-03-30T12:47:09Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 5765501 bytes, checksum: d810d02b334ee8a11679326d3a4f57aa (MD5) Previous issue date: 2015-02-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Ultraviolet radiation (UV) environmental levels have increased in last years and high values are expected until half of this century. Gametes and embryos of marine organisms, whose embryonic development occurs externally, are particularly sensitive to UVA and UVB radiations. The first line of cellular defense against chemical or physical stress is the activity of ABC transporters. The aim of present work was investigate the involvement of ABC transporters on UVA or UVB effects on gametes or embryos of the sea urchin Echinometra lucunter, and investigate the photoactivity and the ABC transporter blocker activity of coumarin and 3-hydroxycoumarin in gametes and embryos of E. lucunter. Gametes or embryos were exposed to UVA (3.6 – 14.4 kJ m-2) or UVB (0.112 – 14.4 kJ m-2), and the embryonic development was monitored by optical microscopy at different developmental stages in the presence or absence of the ABC-transporter blockers reversin205 (ABCB1 blocker) ou MK571 (ABCC1 blocker). E. lucunter eggs, spermatozoa and embryos were resistant to UVA exposure. Resistance to the harmful effects of UVB was strongly associated to ABC transporter activity (embryos > eggs > spermatozoa). ABCB1 or ABCC1 blockage promoted injurious effects on spermatozoa exposed to UVA. ABCC1 transporter blockage increased UVB-dependent damage in eggs while ABCB1 transporter inhibition increased harmful effects of UVB in embryonic cells. ABC transporters blockers also induced abnormalities in larval development derived from spermatozoa or eggs irradiated to UVB. However, ABC transporter activity was not affected by UVB exposure. Additionally, we demonstrate the photoprotective activity of coumarin and 3-hidroxycoumarin against the deleterious effects of UVB on E. lucunter gametes and embryonic cells, and the ABC transporter blocker activity of the 3-hydroxycoumarin. In conclusion, the present study is the first report of the protective role of ABC transporters against UVA and UVB injurious effects in sea urchins gametes and embryonic cells. Furthermore, our results inserts new alternative for the treatment of various diseases related to UVB excessive exposure and also provides alternatives for clinical reversal of multidrug resistance phenotype. / Os níveis ambientais de radiação ultravioleta (UV) têm aumentado significativamente nas últimas décadas e a tendência é que os valores elevados permaneçam até meados deste século. Gametas e embriões de organismos marinhos, cujo desenvolvimento embrionário ocorre no ambiente externo, são particularmente sensíveis às radiações UVA e UVB. A primeira linha de defesa desses organismos contra estresses químicos e físicos é a atividade de efluxo de bombas conhecidas como transportadores ABC. O objetivo do presente trabalho foi investigar o efeito fotoprotetor de transportadores ABC em gametas e embriões do ouriço-do-mar Echinometra lucunter expostos à UVA ou UVB, bem como investigar a fotoatividade e a atividade bloqueadora de proteínas ABC da cumarina e de seus derivados em gametas e embriões de E. lucunter. Os gametas ou embriões foram expostos à UVA (3,6 a 14,4 kJ m-2) ou à UVB (0,112 a 14,4 kJ m-2), na presença ou ausência dos bloqueadores de transportadores ABC: reversina205 (bloqueador de ABCB1); e MK571 (bloqueador de ABCC1). O desenvolvimento embrionário foi monitorado em microscopia óptica comum. Os gametas e zigotos de E. lucunter foram resistentes à UVA. A resistência das células supracitadas à UVB foi fortemente associada à intensidade da atividade dos transportadores ABC (zigotos > óvulos > espermatozoides). O bloqueio dos transportadores ABCB1 e ABCC1 promoveu inibição no desenvolvimento de embriões derivados de espermatozoides expostos à UVA. O bloqueio do transportador ABCC1 aumentou, em 35,9 + 5,8%, os danos aos óvulos expostos à UVB enquanto que o bloqueio de ABCB1 intensificou, em 36,1 + 2,7%, os danos aos zigotos, induzidos pela UVB. Os bloqueadores de transportadores ABC também induziram malformações ao desenvolvimento larval derivado de espermatozoides ou óvulos expostos à UVB. Entretanto, a atividade dos transportadores ABC não foi afetada pela exposição à UVB. Adicionalmente, demonstramos a atividade fotoprotetora da cumarina e seus derivados contra os efeitos deletérios da UVB em gametas e células embrionárias de E.lucunter, e a atividade bloqueadora de transportadores ABC da 3-hidroxicumarina. Em conclusão, o presente estudo é o primeiro relato do papel protetor de transportadores ABC contra os efeitos deletérios da UVA ou UVB em gametas e células embrionárias de ouriços-do-mar. Além disso, esse trabalho também relata, de forma inédita, a atividade fotoprotetora da cumarina e da 3-hidroxicumarina contra a UVB em gametas e embriões irradiados. Estudos posteriores serão necessários para a inserção terapêutica desses compostos para o tratamento de diversas enfermidades relacionadas com a exposição excessiva à UVB.

A acidificação oceânica, resultante do aumento da concentração de CO2 atmosférico, vem alterando a química dos oceanos resultando na diminuição de seu pH. Diversos estudos avaliaram as consequências dessa diminuição no pH oceânico nas taxas de calcificação, reprodução e desenvolvimento em diversos modelos marinhos, entretanto estudos relacionados a outros processos fisiológicos, como a imunidade, e estudos com indivíduos adultos são escassos. Ouriços-do-mar são espécies aderidas ao substrato, importantes para a ciclagem de nutrientes no ambiente marinho, sendo também animais utilizados como bioindicadores para monitoramento ambiental; assim o estudo da resposta imune inata desses animais frente à acidificação dos oceanos é de extrema importância para prever possíveis alterações fisiológicas desses animais e sua capacidade de adaptação. O presente trabalho teve como objetivo avaliar as alterações provocadas pela acidificação oceânica na resposta imune e no balanço ácido base de duas espécies de ouriço-do-mar tropicais: Lytechinus variegatus e Echinometra lucunter durante as estações de verão e inverno; para isso foram analisados os índices fagocíticos, a capacidade de adesão e espraiamento celular dos amebócitos fagociticos além do balanço acido base do liquido celomático após o período de 24 horas e cinco dias de exposição aos pHs 7,6 e 7,3. Foi também avaliada a capacidade de recuperação dessas espécies com o objetivo de verificar se os parâmetros alterados pela exposição conseguiam ser reestabelecidos. Os resultados mostraram que a redução no pH da água do mar alterou a proporção celular, reduziu a capacidade de fagocitose e espraiamento dos amebócitos fagocíticos assim como também afetou o balanço ácido-base do líquido celomático. Foram encontradas diferenças também entre as estações do ano sendo estas encontradas apenas na espécie Lytechinus variegatus. O teste de recuperação mostrou que os parâmetros alterados pela exposição tendem a retornar aos valores controles, mostrando que em curto prazo essas alterações podem não ser irreversíveis, entretanto, mais estudos são necessários principalmente avaliando períodos de exposição prolongados. Juntos nossos resultados mostram que a acidificação oceânica prejudica parâmetros imunes extremamente importantes para a eliminação de patógenos e consequentemente a sobrevivência desses animais em um futuro oceano acidificado. / Ocean acidification due to increased atmospheric CO2 concentration is altering ocean chemistry resulting in the decrease of its pH. Several studies evaluated the effects of this decrease in ocean pH on calcification rates, reproduction and development in different marine models, however studies related to other physiological processes such as immunity and studies with adult animals are scarce. Sea urchins are species adhered to the substrate, important for nutrient cycling in the marine environment, also being used as bioindicators for environmental monitoring. Thus the study of the innate immune response of these animals due to the acidification of the oceans is extremely important to predict possible physiological changes of these animals and their ability to adapt to this condition. This study aimed to evaluate the changes caused by ocean acidification in the immune response and in the acid-base balance of two sea urchin tropical species: Lytechinus variegatus and Echinometra lucunter during seasons of summer and winter; for this, indexes of phagocytosis, cell adhesion and spreading ability were analyzed in addition to the acid-base balance of the coelomic fluid after 24 hours and five days of exposure to pH 7.6 and 7.3The recover ability of these species were also evaluated in order to verify if the parameters altered by exposition could be reestablished. The results shows that a reduction in the seawater pH changed the cell proportion, reduced the ability of phagocytosis and phagocytic amoebocyte spreading as well as affected the acid-base balance of the coelomic fluid. Differences were also found between seasons, but only in the specie Lytechinus variegatus. The recovery test showed that the parameters altered by exposure tend to return to control values, showing that in the short term these changes may not be irreversible, however, further studies are necessary mainly those related to prolonged periods of exposure. Together our results show that ocean acidification impairs immune parameters extremely important for the elimination of pathogens and consequently the survival of these animals in a future acidified ocean.

Planktonic larvae of marine organisms are increasingly being exposed and required to respond to a changing physical environment. Adult sea urchins occupy both intertidal and subtidal waters and broadcast spawn gametes into the water column to contend with variable physical conditions. To answer how populations of invertebrates residing at different depths adequately prepare their offspring to cope with different levels of ultraviolet radiation (UVR), we collected adult purple sea urchins, Strongylocentrotus purpuratus, from four sites (two intertidal and two subtidal (~15 m deep)) on the central coast of CA to compare UV tolerance in offspring. Our measurements of UVA (321–400 nm) and UVB (280–320 nm) irradiance at all collection sites showed UVA and UVB were low or absent in subtidal sites compared to intertidal sites. Our study found that offspring from intertidal populations experience a less severe developmental delay when exposed to environmentally relevant levels of UVR (using artificial lighting) than offspring from subtidal populations. The mean percent cleavage delay for UV-treated embryos relative to the controls was 16% (± 2.3 SE) for intertidal sites and 21.1% (± 2.7 SE) for subtidal sites. This suggests that environmental UV cues or additional environmental cues experienced by intertidal mothers may prepare offspring to resist effects of UV exposure during early development. To further understand differences in biochemistry of the eggs released from mothers of different populations (i.e. differences in maternal investment), we assessed differences in protein abundance among batches of eggs from intertidal and subtidal populations. We identify a range of candidate proteins involved in various cellular processes such as cell cycle regulation, signaling and transport, oxidative stress and metabolism that may help developing embryos cope with UVR stress. These candidate proteins may also help us understand adaptations important for survival of developing marine organisms in changing ocean conditions.

Die Grundlage der Entwicklung multizellulärer Lebewesen bildet die Herausbildung stabiler Zelllinien aus einer einzelnen Zygote. Die Expression spezifischer Kombinationen von Transkriptionsfaktoren ist von zentraler Bedeutung für diesen Prozess. Des Weiteren sind epigenetische Veränderungen des Genoms von Bedeutung, über deren Verhältnis zu Änderungen der Zell-Identitäten weniger bekannt ist. In dieser Arbeit nutze ich Daten aus scATAC-seq und scRNA-seq (mit metabolischer Markierung; scSLAM-seq) um die regulierenden Elemente im Seeigel zu charakterisieren – vom 4-Zell-Stadium, über die Aktivierung des zygotischen Genoms, bis hin zu den Strukturen der frühen Pluteus-Larve. Die Schicksale der einzelnen Zellen des Seeigels sind gut erforscht und ab der vierten Zellteilung etabliert (auch wenn einzelne Veränderungen bis zum 128-Zell Stadium induziert werden können), wonach die Keimblätter fest verankert sind. Plastizität innerhalb der Keimblätter ist mindestens bis zum Ende der Gastrulation möglich. Mithilfe dieser Daten, sowie den bekannten Ergebnissen der Erforschung der Zellschicksale vergangener Jahrzehnte, ist eine Diversifizierung von Zelltypen ersichtlich, die beim 128-Zell-Stadium mit der großen Welle der zygotischen Genomaktivierung (ZGA), einer Veränderung des Chromatinzustandes und dem Verlust der Entwicklungsplastizität einhergeht. Ein kleiner Teil von Genen, im Allgemeinen zelltypspezifische Transkriptionsfaktoren, wird schon vor der großen Welle der ZGA exprimiert, während sich gut offene Chromatinstrukturen auf wenige, distinkte regulatorische Sequenzen beschränken. Von diesem Zeitpunkt an, wird die Genexpression zelltypspezifisch, auch wenn viele Chromatinelemente weiterhin zelltypübergreifend offen sind. Insgesamt sind die Chromatinelemente recht kompakt und Elemente innerhalb der Gene häufig. Danach porträtiere ich noch die regulatorischen Veränderungen die mit der neuralen Entwicklung, sowie der Diversität von skelettbildenden Zellen im Seeigel einhergehen. Zuletzt identifiziere ich ~ 100 Gene, die in die Kalzifizierung des Skelettes des Seeigels involviert sind. / The emergence of multiple, stable cell lineages from a single-cell zygote is the essence of multicellular development. Combinatorial transcription factor expression is central to this process, as well as epigenetic changes whose relationship to changes in cell-identity are far less well understood. In this thesis, I use data from scATAC-seq and scRNA-seq (with metabolic labeling; scSLAM-seq) to characterize the regulatory landscapes of sea urchin development spanning from the 4-cell embryo through maternal zygotic transition (MZT), gastrulation, and the early pluteus larvae with its ecologically relevant structures (~72h). The early fate-map in sea urchins is well understood, providing an ideal model for this analysis; the basic fate-map is established by the fourth cleavage (though inducible lineage changes are possible up to the 128-cell stage) after which germ-layer identity is locked, though there remains considerable plasticity within lineages at least through gastrulation. Using these data, along-side classic research into cell fate maps in the early embryo, I find that cell-type diversification and a loss of plasticity at 128-cell stage corresponds to a major wave of zygotic genome activation (ZGA) and a clear resolution of the chromatin landscape in the sea urchin. However, a subset of genes, often cell-type-specific transcription factors, shows evidence of pre-MZT zygotic expression with early chromatin accessibility limited to few sites with distinct regulatory sequences. From this time, gene expression profiles become highly cell-type-specific, though many regulatory elements remain ubiquitously accessible, suggesting differential transcription factor occupancy at broadly accessible sites. Overall, the regulatory landscape is fairly compact with accessible intragenic elements being frequent. Subsequently, I profile the regulatory changes underlying neurodevelopment and the diversity of skeletogenic cells in the sea urchin. Finally, I also identify ~ 100 genes that are associated with calcification of the sea urchin skeleton.

<p>In light of the paucity of information on the toxicity of lead (Pb), zinc (Zn), copper (Cu) and nickel (Ni) in the marine environment, the aim of this thesis was to generate data on the mechanisms of toxicity of Pb, Zn, Cu and Ni in the early life stages of a very sensitive marine organism, the purple sea urchin (<em>Strongylocentrotus purpuratus</em>). Previous studies in other systems indicated that these metals can impact ionoregulation, especially calcium homeostasis, so an initial study focused on ionoregulatory changes during the first 96h of development under control conditions in embryonic and larval stages of <em>S. purpuratus</em>. Control tests showed that patterns of Ca, K, Na and Mg had an interesting pattern of accumulation over 96 h development and that the most pronounced changes were observed during the gastrulation stage. A variety of biological endpoints were subsequently utilized to examine potential mechanisms of toxicity. Toxicity tests were performed to determine median toxicity threshold values (i.e. EC50: median effective concentration and LA50: median lethal accumulation) for Zn and Pb. Growth, unidirectional Ca uptake rates, whole body ion concentrations (Na, K, Ca, Mg), and Ca ATPase activity were also monitored every 12 h over the first 84 or 96 h of early development to investigate the mechanisms of toxicity during acute and chronic exposures to lethal and sublethal concentrations of Pb, Zn, Cu and Ni in 100% sea water.</p> <p>Sea urchin embryos were very sensitive to Zn with an EC50 of 2.3 µmol/L (95% C.I. = 1.97–2.71 μmol/L) and LA50 of 4.8 (2.16-11.33) μmol/kg. Embryos displayed even higher sensitivity to Pb with an EC50 of 0.36 (0.25-0.49) µmol/L and LA50 of 1.92 (1.67-2.78) μmol/kg. The toxic effects of these metals were increased when tested in combination with extracts of dissolved organic carbon (DOC) from marine and freshwater sources.</p> <p>From studying larvae chronically exposed to Pb (60 µg/L), Zn (139 µg/L), Cu (6 µg/L) and Ni (47 µg/L) it was apparent that these metals rendered their toxic effects, at least in part, through disruption of Ca homeostasis. Unidirectional Ca uptake rates as well as Ca ATPase activity were significantly inhibited at various time points over development, in larvae in these metal exposures. This resulted in significantly lower levels of Ca accumulated in the larvae. Interestingly, larvae showed some capacity for recovery as Ca uptake rates and internal Ca levels returned to control values periodically over development. Also metal initially accumulated in the larvae often returned to control levels at 72 h of development with the exception of Pb, which was the only non essential metal out of the four tested. Metal effects on the whole body levels of three other ions (Na, K, Mg) as well as larval weight were modest. Surprisingly, acute exposure to much higher levels of these same metals at various stages during development had negligible inhibitory effects on unidirectional Ca uptake rate, suggesting a mechanism other than direct competition for the Ca uptake sites. We propose studying the toxicity of contaminants periodically over development as an effective way to detect sub-lethal effects, which may not be displayed at the traditional endpoint of 72 h.</p> / Master of Science (MS)

This study focused on the management of the red sea urchin Loxechinus albus fishery in Chile. The main objective was to design, construct, implement and assess a computer-based simulation model to analyse the biological effects, socio-economic consequences and spatial dynamics resulting from coastal management plans applied to this resource under the system of AMEBR. This was accomplished by using systems dynamics (SD) and geographical information systems (GIS) modelling, in a process of model development, run, optimisation, sensitivity analysis and risk management, and a series of field-based activities carried out at the cove of Quintay. The GIS model developed for allocating sea urchins restocking sites offered a flexible, cost-effective, user-friendly and descriptive technique for support decision-making on management of this species and other benthic resources. Final site selection for restocking was based on the identification, quantification and selection of higher suitability¦availability combinations (site categories). This map showed 16 different suitability¦availability combinations or site categories, ranging from 4¦100 to 8¦100 (suitability points¦availability %). These had an average of 6.44¦69.37 (covering an area of 82.5 Ha overall equivalent to 81.21% of the study area. This site classification demonstrated high heterogeneity between options, and revealed the full variety of alternatives for decision-making. More importantly, the generally high suitability indexes as well as available area emphasised the prospects for restocking sea urchins in this study area. Over and above of the quantitative outcomes obtained from running the GISRM (suitable and available restocking sites) and the BSESM (alternative strategic management plans), the case study-based analysis made it possible to disclose the wider issues related to the red sea urchin coastal management. These results demonstrated the biological inefficiency of traditional size/seasonal restriction-based approach (macro-scenario 1) for sustainable management of the target species. More importantly, final outcomes strongly suggested that a combination of adaptive restocking-based enhancement activities and flexible exploitation constituted a highly attractive approach (macro-scenario 3) for stock management of this fishery in terms of harvestable stock and related incomes. However from the economic analysis, stocking was also found to be economically unfeasible, being a rather cost intensive exercise negatively affected by high natural mortality rates. A single-variable optimisation analysis demonstrated that a higher survival rate is needed to generate sufficient profits to cover major restocking costs and a positive payment, or a cost reduction is essential to make up for the loss. On top to these practical constraints, based on the distinctive modest economic situation prevailing for most Chilean coves and hence their limited capacity to pay for stocking material, unless adequate and constant funding is available to support artisanal associations, they are very unlikely to develop mass release programmes. Given the economic (i.e.: high operating costs) and technical (i.e.: low survival rates) limitations conditioning stocking-based management cost-effectiveness and applicability, wide implementation of mass releases as a major approach for management of the red sea urchin fishery is very unlikely to take place in Chile. This study presents a methodology and offers a tool to design, evaluate and optimise coastal management plans for the red sea urchin in a dynamic, interactive, systematic, integrated and flexible way. The optional strategic management plans proposed on this study may not be applied equally to any AMEBR, as they are the outputs arising from a single cove-specific analysis. Still, the complete methodological framework and analysis procedures developed may be applied to run the BSESM and optimise management of a red sea urchin fishery at any other AMEBR case of study.

The thesis is focused on anatomy and implementation of dissections of 4 selected model invertebrates in school practice. The selected model species include swan mussel (Anodonta cygnea), stick insect (Medauroidea extradentata), cockchafer (Melolontha melolontha) and sea urchin (Echinus esculentus). In case of the cockchafer, the text is based on this model species, but the supporting photographs of dissection are taken from the tropical cetoniid beetle Pachnoda, which has very similar internal anatomy with the cockchafer. For all selected animals, I describe external and internal morphology as well as recommended methods of obtaining the material, proper killing of specimens, and dissection techniques. The text is supplemented with original digital macrophotographs. The thesis includes also an overview of primary and secondary school textbooks with focus on anatomy of the selected animals. The text is supplemented by a glossary of morphological terminology used in this thesis. Attached is a photographic atlas of dissections, which is a combination of this master thesis and my bachelor thesis (defended in 2010), in which I dealt with five model invertebrates in the same way. This is Ascaris suum, Lumbricus terrestris, Helix pomatia, Procambarus aff. fallax and Archimandrita tesselata. The complete...

Field and laboratory studies were used to elucidate the factors affecting temporal and spatial variations of species abundance and structure of macroalgal assemblage and environmental variables between fish farming (FFA) and non-fish farming (NFFA) areas in Hsiao-Lu-Chiao island, a coral island in southwestern Taiwan. Four experiments have been approached: 1. field surveys of macroalgal assemblage structure on 5-m and 10-m depth at 3 sampling sites at FFA (FFA1, FFA2 and FFA3) and 1 sampling site at NFFA from September 2004, January 2005 and April 2005; 2.the relationship between abiotic (monthly maximum air temperature, monthly minimum air temperature, monthly mean air temperature, monthly cumulative precipitation, monthly cumulative irradiance, seawater temperature, light extinction coefficient, water motion, and nutrient (NO3-, NO2-, NH4+, SRP, DON, and DOP) and biotic (seaurchin density) factors and spatio-temporal variations in macroalgal structures analyzed by non-parametric multivariate model; 3. Factors affecting macroalgal abundance and structure: (1). Comparison of growth temperature ranges in different species to field temperature fluctuation; (2).Comparison of growth irradiance ranges in different species to field irradiance fluctuation; 4.Herbivore pressure: (1). Spatio-temporal variations of sea urchin abundance and structure of assemblage; (2). Gut contents and food preference of sea urchin experiment; (3). Herbivore exclusion experiment. Macroalgal %cover, biomass, species richness, diversity (H¡¦) and evenness (J¡¦) showed temporal and spatial variations, low values in January 2005 and also low values in the 5 m- and 10 m-depth areas of FFA1 and the 10 m-depth areas of FFA2. The data of k-dominance curve, hierarchical cluster and ANOSIM tests indicate that macroalgal assemblage is different between 4 sampling sites, between 2 depths and between 3 seasons. Ceratodictyon spongiosum is the most important species that separates September and January assemblages from April assemblage and separates the FFA1 and FFA2 assemblages from the FFA3 and NFFA assemblages. BVSTEP analysis shows that nutrients (NO3-, DON, DOP), temperature, monthly cumulative precipitation, and sea urchin density are the factors corresponding to variations of macroalgal assemblages, this correlation is more significant for 5 m-depth assemblage. Fish farming area FFA1and FFA2 assemblage are affect by sea urchin density, temperature and DON. Sea urchin influnces macroalgal abundance and assemblage structure in FFA1 and FFA2. Macroalgal %cover in 5 m-depth area shows a reversal relationship with sea urchin density; however, this relationship is not observed for 10 m-depth area. FFA1 and FFA2 are belong to high grazing pressure sites as indicated by high sea urchin density and exclusion experiment. Sea urchin gut contents and feeding preference test show that sea urchin has strong food selectivity with Hypnea charoides and Gracilaria coronopifolia as the most preferred species. Herbivore exclusion experiment shows that Hypnea charoides and Gracilaria coronopifolia are the species recruited in the cages. Ceratodictyon spongiosum had high biomass in FFA1 and FFA2 in January, which was ¡¥low DOP/high DON¡¦. The coindicence of temporal variations in FFA3 assemblage structure with a change from ¡¦Halimeda opuntia and Boodlea compostia¡¦ ¡÷ ¡¦Amphiroa fragilissima, Corallina phhulifera and Galaxaura oblongata¡¦ ¡÷¡¦Halimeda opuntia and Boodlea compostia¡¦ with low nitrogen/ high phosphorous¡¦ ¡÷¡¦ high nitrogen/ low phosphorous¡¦ ¡÷ ¡¦low nitrogen/ high phosphorous¡¦ suggest a role of ¡¦low nitrogen (NO3-)/high phosphorous (DOP)¡¦ for FFA3 structure modification. NFFA assemblage is controlled by temperature and monthly cumulative precipitation. Monthly cumulative precipitation in September was higher than January and April, in which Boodlea compostia and Gracilaria coronopifolia were dominant algae in September. The temperature growth responses of algae using the continuous-flow outdoor laboratory tank culture system fit their seasonal growth, reflecting the temperature-dependent manner of seasonal variations in abundance. It could be concluded from the present investigation that the structure of benthic macroalgal assemblage in Hsiao-Lu-Chiao island in southwestern Taiwan is affected by predicted natural and pulse disturbances. Temperature fluctuations involve in overall temporal variations in structure. Sea urchin herbivory and nutrient as pulse nutrient modulate the structure in fish farming area while monthly cumulative precipitation is associated with algal structure in non-fish farming area.

During the Late Pliocene and early Pleistocene, when the paleo-Orinoco delta system transited over the Amacuro Shelf and reached the paleo-shelf-break along the southeastern shoreline of Trinidad. At this time onwards, the shelf-edge delta clinoforms developed further eastward. These deltaic clastic wedges serve as the unique analog in the geological record for an accommodation-driven inner-shelf and shelf-edge delta, developed at an oblique foreland tectonic setting situated at a tropical-equatorial paleogeographic setting. These deposits were influenced by strong Atlantic longshore current, tropical storms, and phytodetrital pulses, and with an exceptionally high sediment accumulation rates. These four aspects make the clastic wedges unique candidates for sedimentological, ichnological, and stratigraphic investigation. The primary objectives of this thesis are to: (a) collect, analyze, and integrate outcrop data on lithofacies, trace fossils, and discontinuity surfaces into a comprehensive depositional and ichnological model for the first growth-fault-guided shelf-marginal pulse of the paleo-Orinoco delta, as recorded in the Mayaro Formation outcrops in southeast Trinidad; and (b) deduce the dominant sedimentary processes during the across shelf transit and their impacts on the benthic infauna as preserved in the Morne L’Enfer Formation outcrops of southwest Trinidad, which are possibly slightly older than the Mayaro Formation. The basal interval of the Morne L’Enfer Formation has specifically been investigated for this purpose, where the deltaic clastic wedges are preserved directly above shelf deposits. The entire Mayaro Formation megasequence is categorized into deposits belonging to twelve different subenvironments based on lithofacies associations and ichnological characteristics. Ichnological evidence indicates that the shelf-edge deltas are one of the most extreme marine environments for benthic metazoans to colonize. However, the combinations and ranking of stress factors affecting the colonizing fauna are diverse and distinct in every individual subenvironment indicating the relative dominances of river-influence, waves, and/or sediment-gravity-flows vis-à-vis slope instability. Due to variations in stress factors, the megasequence also displays dual ichnologic and sedimentologic properties of both the shelf-edge delta lobe(s) and the outer shelf delta lobe(s). A minor transient tidal influence can only be observed in the architectural elements, e.g. elongated interbar embayment and interlobe prodeltaic depocentres, which control topography and enhance tidal effect. Discovery of an unusual monospecific Glossifungites Ichnofacies along an incision surface in the midst of the Mayaro Formation succession enabled a substantial overhaul of the earlier understanding of the formation in terms of its depositional model and stacking pattern. The surface has been re-identified as a canyon/gully cut at the shelf-edge, which possibly acted as a conduit for (a) the mass movements and for (b) the coarse clastic (mostly silt to medium-grained sand) sediment transfer to deep marine settings. The monospecific nature of the Glossifungites Ichnofacies suite indicates that the incision surface was under substantial ecological stresses for the colonizing infauna. The stresses might have arisen from slope instability of the steep canyon/gully walls, mass movements above the incision surface, elevated water turbulence, and lowered salinity from river influx. Five different facies tracts have been identified within the canyon/gully-fill, which crosscuts the shelf-edge delta-front. The facies tracts are dominated by different types of sediment-gravity flow deposits, which are systematically stacked and are almost devoid of trace fossils due to rapid sedimentation rates and slope instability. They are also strikingly different from the surrounding deltaic facies. A high-frequency sequence stratigraphic model involving the influence of growth-fault tectonics on the relative sea-level curve has been invoked to explain the incision of the canyon/gully and its sequential filling processes. On the other hand, the transition from the open shelf to inner-shelf deltaic condition as displayed by the basal members of the Morne L’Enfer Formation is strongly dominated by evidences of river influence with the transient background action of fair-weather waves and storm waves. A peculiar pattern of disappearance of trace fossils produced by irregular sea-urchins highlight that the river influence was quite strong not only at the sediment-water interface but also in the water-column, which affected invertebrate larvae. The initial progradation of the clastic wedge on the shelf was dominated by hyperpycnal flows and waves in contrast to tidal domination in the younger members of the formation.