O presente trabalho tem como objetivo estudar o perfil de acidificação e a inter-relação entre Streptococcus thermophilus TAO, Lactobacillus delbrueckii subsp. bulgaricus LB340, Lactobacillus acidophilus LAC, Lactobacillus rhamnosus LBA, Bifidobacterium lactis BL04 como culturas associadas em leite fermentado. Cinco leites fermentados foram preparados, sendo a composição das co-culturas a variável estudada. O perfil de acidificação foi monitorado e os parâmetros cinéticos, calculados. Os produtos foram submetidos às análises físico-químicas e microbiológicas durante o armazenamento a 4°C. As associações em cultura mista provocaram a redução do tempo de fermentação do leite. Durante os 21 dias de armazenamento o pH e a firmeza dos leites fermentados variaram. Streptococcus thermophilus TAO, Bifidobacterium lactis BL04 e Lactobacillus rhamnosus LBA forneceram contagens acima de 106 log UFC/mL, porém Lactobacillus acidophilus LAC e Lactobacillus bulgaricus LB340 foram inibidos em cultura mista, demonstrando dificuldades de crescimento quando associados às demais bactérias ácido-láticas. / The present study aimed to evaluate the acidification kinectic and inter-relation between Streptococcus thermophilus TAO, Lactobacillus delbrueckii subsp. bulgaricus LB340, Lactobacillus acidophilus LAC, Lactobacillus rhamnosus LBA, Bifidobacterium lactis BL04 like association cultures in fermented milk. Five fermented milks were prepared and studied variable analyzed was the co-cultures composition. Acidification was monitored and the kinectic parameters were calculated. The products were submitted to physical chemistry and microbiological analyses during the storage at 4°C. The associations in mixed cultures promoted the reduction of fermentation time of the milks. During 21 days of storage, pH and firmness of fermented milks varied. Streptococcus thermophilus TAO, Bifidobacterium lactis BL04 and Lactobacillus rhamnosus LBA presented counts above 106 log cfu/mL. However, Lactobacillus acidophilus LAC and Lactobacillus bulgaricus LB340 were inhibited in mixed cultures demonstrating that these strains had difficulty to grow when in associated cultures with lactic acid bacteria.

The presence of annual density banding in certain long-lived reef-building corals provides a record of the coral’s growth rate over time in response to changing environmental conditions. Coral growth is best described by three parameters: linear extension, bulk density, and calcification. Coral growth is generally controlled by the combined influences of light, temperature, and water quality; however, corals are highly responsive to their surrounding conditions and thus record environmental variations through their rates and patterns of skeletal accretion. Because coral growth rates reflect environmental conditions over time, they allow testing of hypotheses regarding the effects of climate change, more specifically global warming which affects sea surface temperatures and rising atmospheric carbon dioxide which affect the aragonite saturation state of seawater. Influences on coral growth include local changes in sea surface temperature and rainfall as well as large scale climatic indices such as the Atlantic Multidecadal Oscillation (AMO), the North Atlantic Oscillation (NAO), and the Southern Oscillation Index (SOI). Chapter 1, Background, reviews the current state of knowledge in three primary areas: 1) coral biology, growth, density band formation, and measurement of extension, density, and calcification, 2) potential climate change impacts on coral growth, and 3) long-term coral growth records. This section is broadly intended to review the literature, identify possible information gaps, and recognize current debate within coral and climate change research. Chapter 2, Sample Size for Coral Sclerochronology, presents data of sample size correlations based on statistical analyses of annual extension rates. A standardized period (1970-1985) of annual extension rates from the largest number of Montastraea faveolata samples available from southeast Florida (136 corals) was used to test correlation on varying spatial scales and to determine sample size requirements for desired levels of correlation based on objective criteria. The results provide basic information on masterchronology construction for sclerochronological growth rate studies and provide a framework from which further growth rate variability can be assessed. Extension and bulk density can be measured from X-ray films of coral skeletal slabs and can be used to calculate calcification. Chapter 3, Relative Optical Densitometry, describes the techniques and associated errors through the process of coral coring, sectioning, X-raying, developing, digitizing, calibrating and analyzing. The principles of relative optical densitometry and the calculation of mass absorption coefficient ratios for aragonite and aluminum standards are explained. Calculated and measured errors are quantified to define the accuracy and precision of these techniques necessary to detect potentially subtle changes in coral growth caused by climate change. Coral cores from the Florida Key, USA, were used to construct growth records over a 60-yr period from 1973-1996. Chapter 4, Coral Growth Records and Climate Change, uses linear extension rate, bulk-density, and calcification rate from annual and sub-annual bands in order to assess: 1) growth averages, variability, and relationships between growth parameters, 2) long term trends with respect to rising carbon dioxide levels and sea surface temperature, 3) correlation with local environmental variables of temperature and rainfall, and 4) correlation with major climate indices of Atlantic Multidecadal Oscillation, North Atlantic Oscillation, and the Southern Oscillation.

Bacterioplankton are keystone organisms in marine ecosystems. They are important for element cycles, by transforming dissolved organic carbon and other nutrients. Bacterioplankton community composition and productivity rates change in surface waters over spatial and temporal scales. Yet, many underlying biological processes determining when, why and how bacterioplankton react to changes in environmental conditions are poorly understood. Here, I used experiments with model bacteria and natural assemblages as well as field studies to determine molecular, physiological and ecological responses allowing marine bacteria to adapt to their environment. Experiments with the flavobacterium Dokdonia sp. MED134 aimed to determine how the metabolism of bacteria is influenced by light and different organic matter. Under light exposure, Dokdonia sp. MED134 expressed proteorhodopsin and adjusted its metabolism to use resources more efficiently when growing with lower-quality organic matter. Similar expression patterns were found in oceanic datasets, implying a global importance of photoheterotrophic metabolisms for the ecology of bacterioplankton. Further, I investigated how the composition and physiology of bacterial assemblages are affected by elevated CO2 concentrations and inorganic nutrients. In a large-scale experiment, bacterioplankton could keep productivity and community structure unaltered by adapting the gene expression under CO2 stress. To maintain pH homeostasis, bacteria induced higher expression of genes related to respiration, membrane transport and light acquisition under low-nutrient conditions. Under high-nutrient conditions with phytoplankton blooms, such regulatory mechanisms were not necessary. These findings indicate that open ocean systems are more vulnerable to ocean acidification than coastal waters. Lastly, I used field studies to resolve how bacterioplankton is influenced by environmental changes, and how this leads to seasonal succession of marine bacteria. Using high frequency sampling over three years, we uncovered notable variability both between and within years in several biological features that rapidly changed over short time scales. These included potential phytoplankton-bacteria linkages, substrate uptake rates, and shifts in bacterial community structure. Thus, high resolution time series can provide important insights into the mechanisms controlling microbial communities. Overall, this thesis highlights the advantages of combining molecular and traditional oceanographic methodological approaches to study ecosystems at high resolution for improving our understanding of the physiology and ecology of microbial communities and, ultimately, how they influence biogeochemical processes.

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.

The aims of the thesis were to document the history of the Cladocera (Crustacea) community structure in the Starolesnianske Lake (the High Tatra Mountains, Slovakia) with regards to environmental changes (e. g., Medieval Warm Period, Little Ice Age, anthropogenic acidification, recovery from acidification), and to analyze in detail the development of zooplankton (Cladocera, Copepoda, Rotifera) during the peaking acidification and recovery from acidification (1978 - 2015). Based on results of both the approaches, the appropriateness of using historical data on zooplankton in Tatra lakes between 1909 - 1913 (Minkiewicz, 1914) as a reference condition for the evaluating biological recovery from acidification should have been assessed. The methods used were paleolimnological analyses of lake sediment (namely subfossil Cladocera), analysis of recent zooplankton, and analyses of lake water chemistry. It was found that in the historical record, relative abundances of Cladocera species significantly changed cca 150 years ago (at the end of the Little Ice Age and, in the main, in the period of anthropogenic acidification). The lake, though, has had a very stable species composition of Cladocera during the last cca 2,000 years, represented by four species: Alona quadrangularis, Alonella excisa, Ceriodaphnia...

Summary: This thesis is focused on collecting information on the importance of dead wood on biogeochemical cycles of chemical elements in the mountain forest ecosystems of central Europe. The goal of this thesis was to determine the concentration of various elements(C, N, P, Ca, Mg, K, Na, Mn, Al, Fe) in the coarse woody debris of spruce samples of different age, stage of decomposition which were collected in different localities in the national park Šumava. Another goal was to determine the speed of which these elements are released from the dead wood. The information on the quantity of different elements and the speed of their release is currently lacking and could lead to better understanding of various biogeochemical cycles of elements in the mountain forest. During the decomposition the concentration of the elements N, P, Al and Fe increased whereas the Kalium concentration decreased. The other elements' (C, N, Ca, Mg, Mn) concentrations varied. Carbon constituted approximately 50% of weight in all of the samples analyzed. We carried out a laboratory experiment based on decomposition of dead wood samples in various stage of decay during a period of six months. The released concentration of CO2 was measured at two to four weeks intervals. The release of CO2 continually decreased during the experiment.....

Sediments of Bohemian Forest lakes are important natural archives. Their sedimentary record covers postglacial history of the lakes as well as history of natural processes in a wider region. It also documents local settlements and changes in landscape management. The lake sediments have attracted the interest of the scientific community since the end of the 19th century. Despite of the fact that modern paleolimnological and paleoecological investigations were already performed in the second half of the 20th century in the Bohemian Forest Mts., the great potential of the lake sediments was not fully utilized in scientific research so far. The ultimate objective of this thesis is to deepen knowledge of these natural archives and support their utilization in future studies. Several specific objectives have been set to achieve the ultimate objective: i) to compare age of the Bohemian Forest lake sediments with the recent knowledge of local deglaciation at the end of the last ice age, ii) to integrate chronostratigraphic marker horizons as one of the tools of Late Glacial sediment dating, iii) to assess the role of bioerosion in chitinous subfossil freshwater invertebrate remains on the record representativeness, and iv) to prove the presumed dystrophic nature of the lakes during the Holocene using...

The aim of this diploma thesis was to describe species composition and biomass of phytoplankton in the Tatra Mountains lakes (Slovakia, Poland) and environmental factors influencing them. The surface layer of 89 lakes was sampled in September 2004. Flagellates from Chrysophyta, Dinophyta and Cryptophyta most frequently dominated the phytoplankton biomass. Lake in the same valley usually differed in their dominant taxonomic group as a result of different lake morphometry, catchment type and detailed geological structure. Based on the level of phytoplankton biovolume, the majority of lakes were ultraoligotrophic and oligotrophic, however, mesotrophic and even eutrophic conditions were recorded in some forest and meadow lakes. An allometric relationship between phytoplankton biomass and chlorophyll a was revealed. Specific chlorophyll content did not differ among taxonomic groups. Altogether 233 species were determined, the most diverse group were Chlorophyta. Species richness of lake was 3-46 and it was negatively correlated with altitude and positively correlated with lake area. The majority of species were rare both in term of biomass and number of lakes occupied by a particular species. Average local species biomass was positively correlated with regional distribution. The similarity of species...

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Interferons (IFNs) are an essential part of innate immunity and contribute to adaptive immune responses. Here, we employed a loss-of-function analysis with human A549 respiratory epithelial cells with a knockout (KO) of the type I IFN receptor (IFNAR KO), either solely or together with the receptor of type III IFN (IFNAR/IFNLR1 KO). The course of rubella virus (RuV) infection on the IFNAR KO A549 cells was comparable to the control A549. However, on the IFNAR/IFNLR1 KO A549 cells, both genome replication and the synthesis of viral proteins were significantly enhanced. The generation of IFN β during RuV infection was influenced by type III IFN signaling. In contrast to IFNAR KO A549, extracellular IFN β was not detected on IFNAR/IFNLR1 KO A549. The bioenergetic profile of RuV-infected IFNAR/IFNLR1 KO A549 cells generated by extracellular flux analysis revealed a significant increase in glycolysis, whereas mitochondrial respiration was comparable between all three cell types. Moreover, the application of the glucose analogue 2-deoxy-D-glucose (2-DG) significantly increased viral protein synthesis in control A549 cells, while no effect was noted on IFNAR/IFNLR KO A549. In conclusion, we identified a positive signaling circuit of type III IFN signaling on the generation of IFN β during RuV infection and an IFN signaling-dependent contribution of glycolysis to RuV infection. This study on epithelial A549 cells emphasizes the interaction between glycolysis and antiviral IFN signaling and notably, the antiviral activity of type III IFNs against RuV infection, especially in the absence of both type I and III IFN signaling, the RuV replication cycle was enhanced.