Ecossistemas aquáticos flutuam em torno de tendências, podendo ocorrer transições súbitas de um regime persistente para outro, alterando a viabilidade dos recursos ou dos parâmetros físico-químicos. Seguindo essa tendência, comunidades variam no tempo e no espaço como resultado de suas interações com o ambiente e com os outros organismos. Comunidades microbianas aquáticas são importantes componentes do metabolismo aquático, atuando na reciclagem de nutrientes pela remineralização e na transferência de biomassa pela alça microbiana. A compreensão da dinâmica funcional microbiana em lagos é uma importante ferramenta para o entendimento desses sistemas, uma vez que a funcionalidade e a composição microbiana podem refletir as condições gerais da lagoa em questão. Desse modo, no presente trabalho, foi utilizada a abordagem de impressões metabólicas da comunidade microbiana na Lagoa Mangueira, uma grande lagoa costeira, rasa e subtropical localizada no extremo sul do Brasil. Teve como objetivo avaliar, pioneiramente para essa região, a diversidade metabólica microbiana aquática através de padrões de consumo de fontes de carbono disponíveis, utilizando Biolog EcoplatesTM e verificar a existência de dinâmicas temporais e espaciais na preferência de consumo desses substratos. Foi observada heterogeneidade temporal e espacial na preferência de consumo de substratos ao longo da Lagoa. Tais preferências puderam ser representadas por diferentes substratos indicadores, associados às estações e aos locais da Lagoa. O consumo das fontes de carbono esteve relacionado com a variabilidade ambiental de fatores como turbidez, transparência da água, nutrientes, clorofila a, carbono orgânico temperatura. Isso evidencia que a dinâmica funcional foi influenciada pela dinâmica de nutrientes, pelos componentes de produtividade, pela sazonalidade e pela compartimentação da Lagoa. Nesse sentido, a investigação do consumo de fontes de carbono, no presente estudo, se mostrou um bom indicador da dinâmica funcional microbiana para ecossistemas aquáticos. / Aquatic ecosystems float around trends in which abrupt transitions can occur between persistents regimes, alterning the viability of the resources or the physical and chemical parameters. Following this trend, communities vary in time and space as a result of their interactions with the environment and other organisms. Microbial communities are important components in the aquatic metabolism, responsible for recycling of nutrients by remineralization, and transfering of biomass through the microbial food web. Understanding the functional microbial dynamics in lakes is an important tool to understand these systems, since the microbial composition and function may reflect the overall condition of the lake. The approach of metabolic fingerprint of microbial communities in shallow lakes was applied in the current study in Lake Mangueira, a costal and large subtropical shallow lake located in southern Brazil. The main goal was to evaluate, pioneered for this region, the heterotrophic microbial metabolic diversity through consumption patterns of available carbon sources using Biolog EcoplatesTM and verify the existence of temporal and spatial dynamics of consumption preference in these substrates. As a result, was observed temporal and spatial heterogeneity of substrate consumption preference among the Lake. Such preferences were represented by different substrate indicators, associated with seasons and sites within the Lake. The substrate utilization was related to environmental variability of factors as turbidity, water transparence, nutrients, chlorophyll a, organic carbon, water temperature. This is evidence that the functional dynamic was influenced by nutrients dynamic, production component, seasonality and compartimentation. In this way, carbon source utilization approach was a good indicator of functional dynamics in the present study to aquatic ecosystems.

<p>Heterotrophic microorganisms are important for the flow of carbon and nutrients in the sea. Bacteria, nanoflagellates and ciliates are relevant components of the pelagic food web. In order to be able to predict the outcome of e.g. eutrophication or climate change we need to know how the different components of the pelagic food web are regulated. With the focus on the northern Baltic Sea food web, this thesis deals with limitation and control of heterotrophic protists, the effect of resource heterogeneity on food web efficiency and diversity of nanoflagellates.</p><p>In-situ microcosm experiments showed that the net growth of heterotrophic flagellates were resource limited throughout the year. Field data confirmed that the abundance of flagellates was bottom-up controlled. Furthermore, field data also showed that the annual average biomass of protists, flagellates and ciliates increased with primary productivity. On a smaller seasonal scale temperature and bacterial biomass were able to explain most of the variation in flagellate biovolume. The temporal variation in ciliate biovolume could not be explained by any bottom-up factors like bacterial biomass, flagellate biomass or chlorophyll a. This and an in-situ microcosm experiment implied that the seasonal dynamics of ciliates were more regulated by predators like mesozooplankton.</p><p>The food web efficiency i.e. how much of production at the resource level is converted to production at the top trophic level, may be affected by specific size or type of resource. Indoor mesocosms revealed that the food web efficiency was 11 times lower when heterotrophic bacteria dominated basal production instead of nano- and micro-sized phytoplankton. This was due to a lengthening of the food web when pico-sized bacteria constituted the main resource.</p><p>The PCR-DGGE molecular biological method was used to study the diversity of heterotrophic or mixotrophic chrysomonads. The focus was set on chrysomonads due to their relatively large contribution to the nanoflagellate community. Group-specific PCR primers were optimized for the target group. A field survey in the northern Baltic Sea showed that a handful of chrysomonad sequences were present throughout the year. Significantly more chrysomonads were recorded in the basin with higher primary productive and salinity. In total 15-16 different chrysomonad sequences were recorded. Most of them matched uncultured chrysomonad clones.</p>

<p>Heterotrophic bacteria utilize dissolved organic matter, and the carbon flow through an ecosystem depends on the fractions of the utilized carbon that is either respired or transferred to higher trophic levels. The major aim this thesis is to investigate 1) the relationship between composition and functioning in heterotrophic bacterioplankton communities and 2) the influence of environmental conditions on both parameters. I set up several batch culture experiments, where lake water filtrates containing bacteria but no grazers were inoculated into sterile freshwater medium to investigate the importance of the origin of the source community (the inoculum) versus the environmental conditions (the medium) for the composition and functional performance of bacterial communities. In some experiments the medium was manipulated to simulate changes in salinity, pH and dissolved organic matter quantity and quality. Functional parameters (biomass yield, respiration, growth efficiency and enzyme activities) and the genetic composition of the emerging bacterial communities were determined.</p><p>When bacterial inocula obtained from different habitats were re-grown under identical conditions, differently composed communities emerged. This indicates that the history and distribution of taxa within the inoculum was an important regulating factor of community composition. The coupling between community composition and functioning was not very tight, and there was functional equivalency with respect to aggregated functions important at the ecosystem scale (e.g., biomass production and respiration). The functional performance of bacterial communities could to a large extent be predicted from the medium alone, except when it deviated strongly from the ambient settings. When bacterial communities were exposed to dilution, a strong change in pH or an increase in salinity, growth of structurally and functionally distinct communities occurred. I therefore suggest that it depends on the disturbance regime how bacterial community structure and function are related to each other. </p>

Le bilan carboné d'une rotation culturale de quatre ans a été établi entre 2004 et 2008 sur le site expérimental de Lonzée, Belgique. La région se caractérise par un climat tempéré océanique avec une température moyenne de l'air de 10°C et des précipitations annuelles de l'ordre de 800 mm. Le sol de la parcelle est un Luvisol. Les cultures composant la rotation sont des cultures de betterave, froment dhiver, pomme de terre et froment dhiver. Dans le but détablir un bilan carboné, des mesures ont été effectuées à différentes échelles spatiales et temporelles. Un système de mesure par eddy covariance fournit une estimation par demiheure de l'échange net en CO2 de l'écosystème (NEE). La qualité des procédures de sélection et de traitement des flux de NEE a été contrôlée. A partir de ces mesures, la productivité primaire brute (GPP) et la respiration totale de l'écosystème (TER) sont déduites. Plusieurs procédures basées sur les mesures diurnes ou nocturnes de NEE sont comparées. L'intérêt d'utiliser un court pas de temps dans ces procédures et la température du sol comme température de référence a été mis en avant, ainsi que la nécessité de déterminer une valeur seuil de la vitesse de friction (u*) pour chaque culture et les longues inter-cultures. Pour la culture de froment d'hiver 2005, une estimation de la GPP déduite des mesures d'eddy covariance est comparée à une estimation basée sur une modélisation de mesures réalisées, une fois par semaine, à l'échelle de la feuille. La conception initiale de l'appareil de mesure a permis de réaliser les mesures sur les feuilles uniquement. Les évolutions des deux estimations sont proches à l'échelle de la saison et à l'échelle journalière. La mise en oeuvre de la méthode basée sur les mesures à léchelle de la feuille a apporté dimportantes informations en termes de réponse de la GPP aux facteurs climatiques et non climatiques et a permis une validation de lestimation basée sur les mesures deddy covariance. Toutefois, dans le cadre de létablissement dun bilan carboné, la méthode basée sur les mesures d'eddy covariance est préférée. Des mesures de la respiration de sol, réalisées à l'échelle de la miniparcelle de sol, et des mesures du contenu en carbone des plantes sont aussi réalisées. Combinées aux mesures d'eddy covariance, ces mesures permettent de déduire les parts autoet hétérotrophiques de la respiration. Sur l'ensemble de la rotation, la parcelle apparait être une source significative de carbone de 0.17 (+/- 0.14) kg C m-2. Cela suggère que, durant la rotation, le contenu en carbone du sol a diminué. Ceci peut sexpliquer par labsence dapport de fertilisation organique durant les 10 dernières années ainsi que par la récolte systématique des pailles des céréales.

Heterotrophic bacteria utilize dissolved organic matter, and the carbon flow through an ecosystem depends on the fractions of the utilized carbon that is either respired or transferred to higher trophic levels. The major aim this thesis is to investigate 1) the relationship between composition and functioning in heterotrophic bacterioplankton communities and 2) the influence of environmental conditions on both parameters. I set up several batch culture experiments, where lake water filtrates containing bacteria but no grazers were inoculated into sterile freshwater medium to investigate the importance of the origin of the source community (the inoculum) versus the environmental conditions (the medium) for the composition and functional performance of bacterial communities. In some experiments the medium was manipulated to simulate changes in salinity, pH and dissolved organic matter quantity and quality. Functional parameters (biomass yield, respiration, growth efficiency and enzyme activities) and the genetic composition of the emerging bacterial communities were determined. When bacterial inocula obtained from different habitats were re-grown under identical conditions, differently composed communities emerged. This indicates that the history and distribution of taxa within the inoculum was an important regulating factor of community composition. The coupling between community composition and functioning was not very tight, and there was functional equivalency with respect to aggregated functions important at the ecosystem scale (e.g., biomass production and respiration). The functional performance of bacterial communities could to a large extent be predicted from the medium alone, except when it deviated strongly from the ambient settings. When bacterial communities were exposed to dilution, a strong change in pH or an increase in salinity, growth of structurally and functionally distinct communities occurred. I therefore suggest that it depends on the disturbance regime how bacterial community structure and function are related to each other.

In aquatic environments the availability of nitrogen (N) generally limits primary production. N2-fixing prokaryotes (diazotrophs) can convert N2 gas into ammonium and provide significant input of N into the oceans. Cyanobacteria are thought to be the main N2-fixers but diazotrophs also include a wide range of heterotrophic bacteria. However, their activity and regulation in the water column is largely unknown. In this thesis the distribution, diversity, abundance, and activity of marine and estuarine heterotrophic diazotrophs was investigated. With molecular methods targeting the nifH gene, encoding the nitrogenase enzyme for N2 fixation, it was shown that diverse nifH genes affiliating with heterotrophic bacteria were ubiquitous in surface waters from ten marine locations world-wide and the estuarine Baltic Sea. Through enrichment cultures of Baltic Sea surface water in anaerobic N-free medium, heterotrophic N2 fixation was induced showing that there was a functional N2-fixing community present and isolates of heterotrophic diazotrophs were obtained. In Sargasso Sea surface waters, transcripts of nifH related to heterotrophic bacteria were detected indicating heterotrophic N2-fixing activity. Nitrogenase expression is thought to be highly regulated by the availability of inorganic N and the presence of oxygen. Low oxygen zones within the water column can be found in association with plankton. The presence of diazotrophs as symbionts of heterotrophic dinoflagellates was investigated and nifH genes related to heterotrophic diazotrophs rather than the cyanobacterial symbionts were found, suggesting that a symbiotic co-existence prevailed. Oxic-anoxic interfaces could also be potential sites for heterotrophic N2 fixation. The Baltic Sea contains large areas of anoxic bottom water. At the chemocline and in anoxic deep water heterotrophic diazotrophs were diverse, abundant and active. These findings extend the currently known regime of N2 fixation to also include ammonium-rich anaerobic waters. The results of this thesis suggest that heterotrophic diazotrophs are diverse and widely distributed in marine and estuarine waters and that they can also be active. However, limits in the knowledge on their physiology and factors which regulate their N2 fixation activity currently prevent an evaluation of their importance in the global marine N budget.

Heterotrophic microorganisms are important for the flow of carbon and nutrients in the sea. Bacteria, nanoflagellates and ciliates are relevant components of the pelagic food web. In order to be able to predict the outcome of e.g. eutrophication or climate change we need to know how the different components of the pelagic food web are regulated. With the focus on the northern Baltic Sea food web, this thesis deals with limitation and control of heterotrophic protists, the effect of resource heterogeneity on food web efficiency and diversity of nanoflagellates. In-situ microcosm experiments showed that the net growth of heterotrophic flagellates were resource limited throughout the year. Field data confirmed that the abundance of flagellates was bottom-up controlled. Furthermore, field data also showed that the annual average biomass of protists, flagellates and ciliates increased with primary productivity. On a smaller seasonal scale temperature and bacterial biomass were able to explain most of the variation in flagellate biovolume. The temporal variation in ciliate biovolume could not be explained by any bottom-up factors like bacterial biomass, flagellate biomass or chlorophyll a. This and an in-situ microcosm experiment implied that the seasonal dynamics of ciliates were more regulated by predators like mesozooplankton. The food web efficiency i.e. how much of production at the resource level is converted to production at the top trophic level, may be affected by specific size or type of resource. Indoor mesocosms revealed that the food web efficiency was 11 times lower when heterotrophic bacteria dominated basal production instead of nano- and micro-sized phytoplankton. This was due to a lengthening of the food web when pico-sized bacteria constituted the main resource. The PCR-DGGE molecular biological method was used to study the diversity of heterotrophic or mixotrophic chrysomonads. The focus was set on chrysomonads due to their relatively large contribution to the nanoflagellate community. Group-specific PCR primers were optimized for the target group. A field survey in the northern Baltic Sea showed that a handful of chrysomonad sequences were present throughout the year. Significantly more chrysomonads were recorded in the basin with higher primary productive and salinity. In total 15-16 different chrysomonad sequences were recorded. Most of them matched uncultured chrysomonad clones.

Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-18T15:32:09Z No. of bitstreams: 1 2013_dis_loleite.pdf: 1004003 bytes, checksum: c14f43a7d45b40582533ad69e13cca53 (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-18T15:32:21Z (GMT) No. of bitstreams: 1 2013_dis_loleite.pdf: 1004003 bytes, checksum: c14f43a7d45b40582533ad69e13cca53 (MD5) / Made available in DSpace on 2016-07-18T15:32:21Z (GMT). No. of bitstreams: 1 2013_dis_loleite.pdf: 1004003 bytes, checksum: c14f43a7d45b40582533ad69e13cca53 (MD5) Previous issue date: 2013 / LEITE, Lana Oliveira. Avaliação do comportamento bacteriano e alterações sensoriais em tilápias do Nilo (Oreochromis niloticus) durante estocagem em gelo. 2013. 59 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Engenharia de Pesca, Fortaleza-CE, 2013 / Fish is a food susceptible to deterioration due their intrinsic characteristics, requiring bacterial monitoring in postmortem storage. Thus, the objective was to quantify mesophilic, psychrotrophic, psychrophilic bacteria in Nile tilapia (Oreochromis niloticus) during to 21 days of storage under the action of ice at temperature refrigeration. Four experiments were conducted, separated to two groups, gutted and ungutted, with eight samples totaling 32 samples. Every three days was taken a exemplar of tilapia for analyzes of the trial: sensory evaluation according to the Quality Index Method (QIM) adapted to the Nile tilapia and heterotrophic bacterial count of cultivable. In sensory analysis of raw fish attributes were assessed: general appearance, eyes, gills, muscles and texture of the fish, according to the Sensory Characters Sum (S.C.S). To each specimen were quantified bacterial groups: psychrophilic (23 °C), psychrotrophic (7 °C) and mesophilic (35 °C), the surface and the muscle, using the method of standard plate count (SPC). There was no statistical difference between the three bacterial groups analyzed the significance level of 5% within the same treatments and between groups. The storage time was highly correlated with the counting of bacterial groups and increased QIM for both eviscerated fish, and for those not drawn. It was considered that the process of evisceration on the premises of the fish did not increase its useful life. It is suggested, therefore, greater attention in the process of gutting fish markets that sell live fish, because this process was not carried out effectively to security and recovery product. / O pescado é um alimento susceptível à deterioração devido às suas características intrínsecas, sendo necessário o monitoramento bacteriano no armazenamento pós-morte. Dessa forma, objetivou-se quantificar bactérias, psicrotróficas, psicrófilas e mesófilas em tilápias do Nilo (Oreochromis niloticus) durante 21 dias de estocagem sob ação do gelo. Foram realizados quatro experimentos, separados em dois grupos, não eviscerados e eviscerados, cada um com oito amostras perfazendo um total de 32 amostras. A cada três dias era retirado um exemplar de tilápia para as análises do experimento: sensorialmente, conforme a avaliação do Método do Índice de Qualidade (MIQ), adaptado à tilápia do Nilo e microbiologicamente. Na análise sensorial do peixe cru foram avaliados os atributos: aspecto geral, olhos, brânquias, textura e musculatura dos peixes, segundo os somatórios dos caracteres sensoriais (S.C.S). De cada exemplar foram quantificados os grupos bacterianos: psicrotróficos (7°C), psicrófilos (23°C), e mesófilos (35°C), na superfície e no músculo, pelo método de Contagem Padrão em Placas (CPP). Não houve diferença estatística entre os três grupos bacterianos analisados à um nível de significância de 5% dentro dos mesmos tratamentos e entre os grupos. O tempo de estocagem apresentou alta correlação com a contagem dos grupos bacterianos e o aumento do MIQ, tanto para os peixes eviscerados, quanto para os não eviscerados. Foi considerado que o processo de evisceração no local de venda dos peixes não aumentou sua vida útil. Sugere-se, portanto, uma maior atenção no processo de evisceração em peixarias que comercializam peixes vivos, pois esse processo não foi realizado de forma eficaz para segurança e valorização do produto.

Ecossistemas aquáticos flutuam em torno de tendências, podendo ocorrer transições súbitas de um regime persistente para outro, alterando a viabilidade dos recursos ou dos parâmetros físico-químicos. Seguindo essa tendência, comunidades variam no tempo e no espaço como resultado de suas interações com o ambiente e com os outros organismos. Comunidades microbianas aquáticas são importantes componentes do metabolismo aquático, atuando na reciclagem de nutrientes pela remineralização e na transferência de biomassa pela alça microbiana. A compreensão da dinâmica funcional microbiana em lagos é uma importante ferramenta para o entendimento desses sistemas, uma vez que a funcionalidade e a composição microbiana podem refletir as condições gerais da lagoa em questão. Desse modo, no presente trabalho, foi utilizada a abordagem de impressões metabólicas da comunidade microbiana na Lagoa Mangueira, uma grande lagoa costeira, rasa e subtropical localizada no extremo sul do Brasil. Teve como objetivo avaliar, pioneiramente para essa região, a diversidade metabólica microbiana aquática através de padrões de consumo de fontes de carbono disponíveis, utilizando Biolog EcoplatesTM e verificar a existência de dinâmicas temporais e espaciais na preferência de consumo desses substratos. Foi observada heterogeneidade temporal e espacial na preferência de consumo de substratos ao longo da Lagoa. Tais preferências puderam ser representadas por diferentes substratos indicadores, associados às estações e aos locais da Lagoa. O consumo das fontes de carbono esteve relacionado com a variabilidade ambiental de fatores como turbidez, transparência da água, nutrientes, clorofila a, carbono orgânico temperatura. Isso evidencia que a dinâmica funcional foi influenciada pela dinâmica de nutrientes, pelos componentes de produtividade, pela sazonalidade e pela compartimentação da Lagoa. Nesse sentido, a investigação do consumo de fontes de carbono, no presente estudo, se mostrou um bom indicador da dinâmica funcional microbiana para ecossistemas aquáticos. / Aquatic ecosystems float around trends in which abrupt transitions can occur between persistents regimes, alterning the viability of the resources or the physical and chemical parameters. Following this trend, communities vary in time and space as a result of their interactions with the environment and other organisms. Microbial communities are important components in the aquatic metabolism, responsible for recycling of nutrients by remineralization, and transfering of biomass through the microbial food web. Understanding the functional microbial dynamics in lakes is an important tool to understand these systems, since the microbial composition and function may reflect the overall condition of the lake. The approach of metabolic fingerprint of microbial communities in shallow lakes was applied in the current study in Lake Mangueira, a costal and large subtropical shallow lake located in southern Brazil. The main goal was to evaluate, pioneered for this region, the heterotrophic microbial metabolic diversity through consumption patterns of available carbon sources using Biolog EcoplatesTM and verify the existence of temporal and spatial dynamics of consumption preference in these substrates. As a result, was observed temporal and spatial heterogeneity of substrate consumption preference among the Lake. Such preferences were represented by different substrate indicators, associated with seasons and sites within the Lake. The substrate utilization was related to environmental variability of factors as turbidity, water transparence, nutrients, chlorophyll a, organic carbon, water temperature. This is evidence that the functional dynamic was influenced by nutrients dynamic, production component, seasonality and compartimentation. In this way, carbon source utilization approach was a good indicator of functional dynamics in the present study to aquatic ecosystems.

Green algae are quite important primary producers in fresh waters. The genus Chlorella represents one of algae most frequently utilized in algal biotechnologies to produce biomass, using either autotrophic or heterotrophic cultivation systems. It is than exploited as a food supplement for humans or animals. However, particular species within the genus are morphologically indistinguishable and molecular markers should be used to characterize production strains. This work is aimed to molecularly characterize three production strains of Chlorella for patent protection purposes and to specify their phylogenetic and taxonomic position.