A high-resolution record of organic-walled dinoflagellate cyst production in Guaymas Basin, Gulf of California (Mexico) reveals a complex paleoceanographic history over the last ~40 ka. Guaymas Basin is an excellent location to perform high resolution studies of changes in Late Quaternary climate and paleo-productivity because it is characterized by high primary productivity, high sedimentation rates, and low oxygen bottom waters. These factors contribute to the deposition and preservation of laminated sediments throughout large portions of the core MD02-2515. In this study we document dinoflagellate cyst production at a centennial to millennial scale throughout the Late Quaternary. Based on the cyst assemblages three major dinoflagellate cyst zones, with seven subzones were established. The most dominant dinoflagellate cyst taxa found throughout the core were Brigantedinium spp. and Operculodinium centrocarpum. Dansgaard-Oeschger events 5-8 are inferred in the dinoflagellate cyst records on the basis of increases in warm taxa, such as Spiniferites pachydermus. Preceding and during the Last Glacial Maximum cysts of Polykrikos cf. kofoidii increase in abundance, responding to oceanographic changes in the Gulf of California perhaps caused by a regression in sea-level. Other intervals of interest are the Younger Dryas where cooler conditions are not recorded, and the Holocene which is characterized by the consistent presence of warm water species Stelladinium reidii, Tuberculodinidum vancampoae, Bitectatodinium spongium and an increase in Quinquecuspis concreta. Changes in cyst assemblages, concentrations and species diversity, along with geochemical data reflect major millennial scale climatic and oceanographic changes. / Graduate

Dinoflagelados foram os primeiros microorganismos reconhecidos como fonte primária de certos metabólitos, também conhecidos como ficotoxinas. Em geral são compostos não protéicos, de baixo peso molecular (250-3500 Da), apresentam estruturas químicas e modos de ação bem diferenciados. Envolvidos em florações, mortandade de peixes e intoxicações de seres humanos, devido ao consumo de mariscos contaminados, tornaram alvo de diversas pesquisas. Atividade antifúngica, hemolítica e citotoxicidade contra células tumorais são alguns exemplos da variedade de compostos e modos de ação que podemos encontrar. Como exemplo do potencial de aplicação, a pectenotoxina é candidata a agente quimioterápico e a maitotoxina é um estimulante da síntese e secreção do fator de crescimento. Em alguns casos, estudos com compostos de dinoflagelados alcançam níveis de patente. O risco à saúde pública e as perdas econômicas causadas pelas florações levaram ao estudo do ciclo de vida e interações ecológicas, taxonomia e aspectos moleculares das microalgas envolvidas nesses eventos. Também levou a criação de programas de monitoramento que levam em consideração observações ambientais, amostras de fitoplâncton, mariscos ou peixes, análise das amostras, avaliação e divulgação dos resultados e planos de ação em caso de eventos tóxicos. Considerando os estudos já realizados com dinoflagelados, estudamos algumas espécies isoladas do litoral norte do Estado de São Paulo sob aspectos biológicos, moleculares, químicos e monitoramento. A abordagem biológica revelou a presença de compostos capazes de inibir a proliferação celular e de alterar de alguma forma o citoesqueleto das células tumorais T47D nos extratos polares de A. fraterculus, P. gracile e P. mexicanum. Observamos também um possível potencial genotóxico das espécies A. fraterculus e P. gracile. Os aspectos moleculares, biológicos e químicos foram utilizados como ferramenta para o estudo de uma espécie de Ostreopsis cf ovata. A biologia molecular associada a morfologia nos levou a identificar duas cepas, mostrando que são geneticamente idênticas entre si e entre os espécimes isolados do Rio de Janeiro e Mediterrâneo e diferente daqueles de origem asiática, contribuindo para a compreensão da taxonomia, biogeografia e filogenia deste gênero. Ambas as cepas mostraram atividade hemolítica, mas as frações obtidas por cromatografia líquida de alta eficiência não tiveram a hemólise inibida por ouabaína. A análise por espectrometria de massa também não foi conclusiva devido à falta de padrão da toxina estudada. Portanto, sugerimos que os compostos hemolíticos presentes nas amostras sejam distintos da palitoxina e de seus análogos. No entanto, analisando os cromatogramas de espectrometria de massa, verificamos que a atividade biológica encontrada para as nossas amostras estão relacionadas a um conjunto de compostos. O programa de monitoramento piloto na maricultura da Praia da Cocanha em Caraguatatuba, litoral Norte do Estado de São Paulo foi realizado durante um ano. As análises de toxinas paralisantes (PSP) em bioensaios com camundongos e espectrometria de massa não detectaram toxinas nos mexilhões em concentrações que poderiam provocar intoxicação ao homem. Mas confirmamos o alerta quanto à presença de espécies potencialmente tóxicas no ambiente durante o período de estudo. Os aspectos abordados mostram como os compostos produzidos por dinoflagelados podem interferir de alguma forma na vida humana. Por um lado encontramos as finalidades farmacológicas na busca de novas drogas por outro o risco à saúde através do consumo de mariscos contaminados. Em ambos os casos, a continuidade das pesquisas trará muitos benefícios ao Homem. / Dinoflagellates were the firs microorganisms recognized as a primary source of certain metabolites, also known as phycotoxins. In general, compounds are non proteic, with low molecular weight (250-3500 Da), and have different chemical structures and modes of action. Frequently involved in algal blooms, killing fish and poisoning of humans due to consumption of contaminated seafood, they became the target of several researchers. Antifungal, hemolytic and cytotoxic activities against tumor cells are representative of the variety and modes of action that can be found. As examples of potential applications, the pectenotoxin is considered promising candidate for cancer chemotherapy; and the maitotoxin is described as stimulant of the synthesis and secretion of growth factor. In some cases, dinoflagellates compounds reached patent levels. The need to protect human health and fisheries industry led to studies of life cycle, ecologic interactions and taxonomic and molecular aspects of microalgae involved in harmful blooms. These events led to the establishment of monitoring programs to detect phytotoxins and toxic microalgae in mariculture farms worldwide. Based on researches carried out with others dinoflagellate, we studied some species isolated in the North coast of the state of São Paulo, on biological, molecular, chemical and monitoring aspects. Biologic assays indicated the presence of compounds in the dinoflagellates extracts that are able to inhibit cell proliferation and to induce changes in T47D cells cytoskeleton. We suggest a genotoxic potential for the compounds in the polar extracts of A. fraterculus and P. gracile. Biologic, molecular and chemical aspects were used to study two strains of Ostreopsis cf ovata. Molecular biology showed that both are genetically identical to O. ovata isolated from the Mediterranean Sea and Rio de Janeiro, but different from O. ovata collected in Asian waters. This result allowed a better understanding of genetic diversity and the recognition of more reliable (stable) morphological characters. Using mouse erythrocytes assay, hemolytic activity was detected in both strains of O. cf ovata. However, the hemolytic component was not inhibited by ouabain. Analyses by mass spectrometry of polar extract were inconclusive due to the lack of toxins standards. Therefore, we suggest that the compounds in the samples are distinct from palytoxin. Mass spectrometry results indicate that the biological activity of our samples can be related to several substances. The monitoring program in the mussel farm at Cocanha Beach, Caraguatatuba, was carried during one year. Mouse bioassays for paralytic shellfish poisoning toxins (PSP) performed in mussels showed that concentrations were below regulatory levels for human consumption. These low levels of PSPs were also confirmed by mass spectrometry. However, potentially toxic species were detected, and a long-term monitoring program is proposed and highly recommended. Research shows that compounds produced by dinoflagellates can interfere in human life both in positive and negative ways. On one side is the search for new drugs for pharmacological purposes; on the other hand the health risk represented by the consumption of contaminated seafood. In both cases, the continuity of the research will bring many benefits to humans.

Following their endosymbiotic uptake, plastids undergo profound changes to genome content and to their associated biochemistry. I have investigated how evolutionary transitions in plastid genomes may impact on biochemical pathways associated with plastid gene expression, focusing on the highly unusual plastids found in one group of eukaryotes, the alveolates. The principal photosynthetic alveolate lineage is the dinoflagellate algae. Most dinoflagellate species harbour unusual plastids derived from red algae. The genome of this plastid has been fragmented into small, plasmid-like elements termed “minicircles”. Transcripts of this genome receive a 3’ poly(U) tail and, in some species, undergo extensive sequence editing. Some dinoflagellates have replaced their original plastids with others, in a process termed “serial endosymbiosis”. The major non-photosynthetic alveolates are the apicomplexans, which include the malaria parasite Plasmodium. Apicomplexans are descended from free-living algae and possess a vestigial plastid, which originated through the same endosymbiosis as the ancestral red dinoflagellate plastid. This plastid has lost all genes involved in photosynthesis and does not possess a poly(U) tail addition pathway. I have investigated the consequences of the fragmentation of the red algal dinoflagellate plastid genome on plastid transcription. I have characterised non-coding transcripts in plastids of the dinoflagellate Amphidinium carterae, including the first evidence for antisense transcripts in an algal plastid. Antisense transcripts in dinoflagellate plastids do not receive poly(U) tails, suggesting that poly(U) tail addition may play a role in strand discrimination during transcript processing. I have additionally characterised transcript processing in dinoflagellate plastids that were acquired through serial endosymbiosis. I have shown that poly(U) tail addition and editing occur in the haptophyte-derived serial endosymbionts of the fucoxanthin-containing dinoflagellates Karenia mikimotoi and Karlodinium veneficum. This is the first evidence that plastids acquired through serial endosymbiosis may be supported by pathways retained from previous symbioses. Transcript editing constrains the phenotypic consequences of divergent mutations in fucoxanthin plastid genomes, whereas poly(U) tail addition plays a central role in recognising and processing translationally functional fucoxanthin plastid mRNAs. I have additionally shown that certain genes within fucoxanthin plastids are located on minicircles. This demonstrates convergent evolution in the organisation of the fucoxanthin and red algal dinoflagellate plastid genomes since their endosymbiotic acquisition. Finally, I have investigated transcript processing in the algae Chromera velia and Vitrella brassicaformis. These species are closely related to apicomplexans but are still photosynthetic and apply poly(U) tails to plastid transcripts, as with dinoflagellates. I have shown that poly(U) tails in these species are preferentially associated with translationally functional mRNAs of photosynthesis genes. This is the first plastid transcript processing pathway documented to target a specific functional gene category. Poly(U) tail addition may direct transcript cleavage and allow photosynthesis gene transcripts to accumulate to high levels. The loss of this pathway from ancestors of apicomplexans may have contributed to their transition from photosynthesis to parasitism.

In situ hybridization (ISH) has played a crucial role in developing a spatial transcriptomic understanding of emerging model organisms in the past, but advancing high-throughput RNA-sequencing (RNA-seq) technology has pushed this method into the shadows, leading to a loss of data resolution. This shift in research towards the exclusive use of RNA-seq neglects essential considerations for transcriptomic studies including the spatial and temporal expression of transcripts, available budget, experimental design needs, and validation of data. A synergy of spatiotemporal transcriptomic techniques is needed, using the bulk and unbiased analysis of RNA-seq and the visual validation and spatiotemporal resolution of ISH. Integration of this synergistic approach can improve our molecular understanding of non-model organisms and establish the background data needed for advancing research techniques. A prime example lies within an emerging model of the marine science and symbiosis fields, where I present a case study on a threatened coral reef keystone – the cnidarian-dinoflagellate symbiosis. Establishing a whole-mount ISH protocol for the emerging cnidarian model Aiptasia (sea anemone) will help future studies reveal the gene regulation underpinning the establishment, persistence, and breakdown of this complex symbiotic relationship.

Dynamic Systems modeling suggests that complex coastal dinoflagellate biosystems can be modeled using environmental parameters such as temperature, salinity, and bulk nutrient levels. The former Salisbury Embayment of northern Virginia and Maryland is modeled here, using STELLA I and FORTRAN models based on physical oceanography and temperature, salinity, and nutrient conditions of the modern Yellow Sea. In these models, dinoflagellate assemblages are predicted based on environmental conditions associated with depth. Cluster analyses of fossil dinoflagellate frequency data from Tertiary Pamunky Group (Aquia and Nanjemoy Formations) of the Salisbury Embayment produce 17 discrete groupings. Samples within the Salisbury fossil cluster groups are statistically similar (via ANOVA analysis), but not the same. Therefore they represent paleocommunity types rather than paleocommunities. Although individual sinofiagellate species recur in similar environmental settings, the paleocommunity types do not appear to repeat. In the past, such associations have been used as depth indicators. It is suggested here that they relate to estuarine, nearshore, and offshore coastal regions because of the temperature, salinity, and nutrient conditions of each. In the modern Yellow Sea, nearshore and offshore regions are separated by discrete lateral fronts in some areas, and by gradational regions of mixing in others. Both types of water mass boundaries are modeled in this study. Results suggest that evidences that discrete water mass boundaries might have occurred between some fossil dinoflagellate associations. Circulation patterns of the Salisbury Embayment may have been roughly similar to those of the modern Yellow Sea. / Master of Science

To study the ecology of organic-walled dinoflagellate cysts, the changes in species composition, diversity, and seasonal variations of cyst flux in the sediment trap deployed in the Strait of Georgia (BC, Canada) were examined. The cyst production rate varied from ~600 to 336,200 cysts m-2 day-1, with an average of 20,000 cysts m-2 day-1. Throughout the study period (March, 1996 - January, 1999), cyst assemblages were mostly dominated by cysts produced by heterotrophic dinoflagellates, such as Protoperidineaceae (Brigantedinium spp., Quinquequspis concreta, and cysts of Protoperidinium americanum). Cysts produced by heterotrophic dinoflagellates peaked in June each year, whereas cysts produced by autotrophic taxa were most abundant during August-September. The total annual dinoflagellate cyst flux was higher in 1996 than in 1997 and 1998, being enhanced by the bloom of Alexandrium spp. The warmer sea-surface temperature in 1998 had a positive effect on the production of both autotrophic and heterotrophic dinoflagellates, as inferred from the cyst fluxes.

The species composition of planktonic communities is determined not only by abiotic factors, such as nutrient availability, temperature, and water column stratification but also by biotic interactions between hosts and parasites, predators and prey, and among competitors. Blooms of the red tide dinoflagellate, Karenia brevis, can dramatically alter the planktonic community, reaching densities of millions of cells per liter and occurring nearly monospecifically. I investigated whether K. brevis uses chemical compounds to defend against grazing or to inhibit the growth of competitors. Because K. brevis is known to produce brevetoxins which act as potent neurotoxins in mammals, I also investigated whether brevetoxins played a role in competition or predator resistance. Experiments revealed that copepods fed diets rich in Karenia brevis experienced lowered fitness, however, nutritional inadequacy, rather than toxicity, was responsible for the decrease in grazer fitness. Compounds exuded from natural samples of K. brevis blooms did, however, inhibit the growth of four of five model competitors. Compounds exuded from K. brevis cultures were similarly allelopathic to competitors. Exposure to these allelopathic compounds resulted in lowered photosynthetic efficiency of all competitors, and decreased cell membrane integrity of three competitors. The allelopathic potency of K. brevis blooms was variable between collections and years, but allelopathy did not correlate with bloom density or concentration of brevetoxins. However, the variability of allelopathy could partially be explained by the presence of specific competitors. The diatom Skeletonema costatum reduced the growth-inhibiting effects of K. brevis bloom exudates, suggesting that S. costatum has a mechanism for undermining K. brevis allelopathy. Allelopathic compounds exuded by K. brevis that inhibited the growth of the diatom Asterionellopsis glacialis were partially characterized. K. brevis produced multiple, polar, organic compounds that inhibited A. glacialis growth. Exuded brevetoxins, on the other hand, had no effect on A.glacialis growth. Taken together, these results indicate that K. brevis is not chemically defended against grazing, but does produce yet-unidentified allelopathic compounds that inhibit the growth of competing phytoplankton. Blooms of K. brevis may be facilitated by the exudation of potent allelopathic compounds, but the specific phytoplankton assemblage has the potential to alter bloom dynamics.

Thesis (Dr. rer. nat.)--Universität Tübingen, 2004. / Includes bibliographical references. Available in PDF format via the World Wide Web.

No description available.

This study contributes to developing terrestrial and marine palynological indicators of winter or summer monsoon signals as well as oceanographic environments of the South China Sea (SCS). The high-resolution reconstructions of Holocene East Asian Monsoon (EAM) climate and oceanographic condition of the northern SCS provide insights into regional climate events in the western low-latitude Pacific Ocean and their impacts on local oceanography and ecology. Sediment trap samples from the southwest Taiwan waters of the SCS in winter monsoon (March-April) and summer monsoon (July-August) seasons identify abundances of Pinus and Ulmus pollen as indicators of the winter monsoon whereas fern spores appeared to be indicators of the summer monsoon. The increased fluxes of dinoflagellate cyst (DC) taxa during summer are correlated with decreased sea-surface salinity (SSS) associated with nutrient-rich river inputs. DC distributions across the SCS show that some taxa are good indicators of changes in sea-surface temperature (SST), SSS, water depth and chlorophyll-a (chl-a) concentrations associated with EAM and oceanographic conditions. In particular, the concenrations of Brigantedinium spp. and cysts of Protoperidinium together with Echinidinium spp. are positively correlatd with SST in January and SST in July, and chl-a concentrations, respectively, which are linked to past monsoon strength and primary productivity. In total, four high cyst concentration regions have been observed off southern Vietnam, Borneo, Hainan, and South China. High-resolution palynological records from a sediment core in the northern SCS reflect several EAM climatic and oceanographic events over the last 12.5 kyr. A short-term Impagidinium decrease implied that the Taiwan Strait opened at ~11.7–11.0 cal kyr BP, with reduced Kuroshio Current influence when the East China Sea waters entered through the strait. Three Holocene relative sea-level stages were identified in the palynomorph records. The highest herb pollen abundances were observed before ~10.4 cal kyr BP, reflecting the shortest distance from the grassland sources on the exposed shelf at the low sea-level stand. High Brigantedinium and cysts of Protoperidinium abundances also indicate a near-shore environment. During ~10.4- ~6.8-6.0 cal kyr BP at the rising sea-level stage, fern spore abundances increased and DC abundances decreased. Consistently low total DC concentrations and high fern spore abundance were observed after ~6.8-6.0 cal kyr BP when the present oceanographic conditions were formed. Increased abundances of Pinus pollen reflected three strengthened winter monsoon intervals at ~5.5, 4.0 and 2.5 cal kyr BP under the present oceanographic conditions. The highest Dapsilidinium pastielsii abundances reflected the warmest interval at ~6.8-5.5 cal kyr BP of the northern SCS. / Graduate / 2019-12-13