This thesis investigates the effects of co-culturing microorganisms including 37 yeast, 38 bacteria, nine diazotrophic cyanobacteria, and three fungi on biomass and lipid production in fresh- and saltwater algae. Algal lipid content was measured using Nile Red method and gravimetric techniques. Among the algal strains tested, freshwater Coelastrum sp. 46-4, and saltwater Cricosphaera sp. 146-2-9, showed enhanced biomass yield and lipid content in response to co-culture with bacteria, cyanobacteria, and fungi. While co-culture with yeast caused inhibition of algal productivity, no difference in algal productivity was observed between nitrogen-free diazotrophic cyanobacterial co-culture and nitrogen-replete monoalgal culture. Results indicated that extracellular compounds from the freshwater bacteria Pseudomonas stutzeri and marine fungus Fusarium sp. significantly account for stimulation of lipid accumulation within algal cells, while co-cultivation with live microorganism cells stimulated biomass production in algae.

Oceans are powered by active, metabolically diverse microorganisms, which are important in regulating biogeochemical cycles on Earth. Most of the ocean surface is often limited by nutrients, influencing bacterial growth and activities. Bacterial adaptation to fluctuating environmental conditions involves extensive reprogramming, and redirection of bacterial metabolism and physiology. In this thesis, I investigated the molecular mechanisms of bacterial adaptation strategies to sustain their growth and survival, focusing on the regulation of gene and protein expression in heterotrophic marine bacteria. Comparative proteomics analyses of the growth and non-growth conditions, uncovered central adaptations that marine bacteria employ to allow them to change their metabolism to support exponential growth in response to nutrients and to readjust to stationary phase under nutrient limitation. Our results highlight that during nutrient rich conditions three distinct bacteria lineages have great similarities in their proteome. On the other hand, we observed pronounced differences in behavior between taxa during stationary phase. Analyses of the proteorhodopsin containing bacterium Vibrio sp. AND4 during starvation showed that significantly improved survival in the light compared to darkness. Notably, proteins involved in promoting cell vitality and survival had higher relative abundance under light. In contrast, cells in the dark need to degrade their endogenous resources to support their basic cellular demands under starvation. Thus, light strongly influences how PR-containing bacteria organize their molecular composition in response to starvation. Study of alternative energy generation metabolisms in the Alphaproteobacteria Phaeobacter sp. MED193 showed that the addition of thiosulfate enhanced the bacterial growth yields. Concomitantly, inorganic sulfur oxidation gene expression increased with thiosulfate compared to controls. Moreover, thiosulfate stimulated protein synthesis and anaplerotic CO2 fixation. These findings imply that this bacterium could use their lithotrophic potential to gain additional energy from sulfur oxidation for both improving their growth and survival. This thesis concludes that analyses in model organisms under defined growth conditions gives invaluable knowledge about the regulatory networks and physiological strategies that ensure the growth and survival of heterotrophic bacteria. This is critically important for interpreting bacterial responses to dynamic environmental changes. Moreover, these analyses are crucial for understanding genetic and proteomic responses in microbial communities or uncultivated organisms in terms of defining ecological niches of planktonic bacteria

No description available.

In recent decades the quality and safety of bottled water has come into question, while bottled water sales and demand have steadily grown. It is important that consumers as well as manufacturers are made aware of the microbial environment of bottled water. Many studies have been implemented to evaluate the microbiological content of bottled water. Although some laboratory studies have shown that microbial counts of bottled water can reach as high as 16^3 CFU/mL, few studies, if any, have studied the impact of storage location on bottled water microbial activity. This document reports on an investigation that evaluated the microbiological quality of bottled water relative to storage conditions and storage duration. Unlike previous studies that evaluated bottled water having been stored under the laboratory conditions, the work conducted in this study evaluated storage scenarios that included a car trunk, covered porch, indoor cabinet and refrigerator. These storage conditions allow for comparison of prior studies conducted in the laboratory to more realistic storage coniditions used by consumers. Analyses of bottled water under these alternative storage conditions indicated that microbial growth did occur in stored water and varied between lcoation and holding time. It was determined that heterotrophic plate counts (HPCs) were greater in warmer storage environments as was exhibited by the refrigerated and indoor locations. Additionally, mathematical models were developed in this work that predicted the microbial growth rate in bottled water as a function of holding time, using commonly available statistical software that evaluated data predicted using an exponential model (R2 correlation of up to 0.84) for two different storage conditions. Although increased levels of HPC bacteria are generally safe for those in good health, they are used as an indicator test for microbial quality. Furthermore, higher levels of HPC have also been shown to pose some additional health risks to immunocomprimised individuals. Therefore, results from the study would indicate that it may be beneficial for consumers to store their bottled water indoors or in a refrigerator.

No description available.

Fertilization is a common, cost effective treatment for increasing forest productivity within managed forests of the southeastern United States. However, little is known about how fertilization affects the below-ground processes that drive soil CO2 efflux in loblolly pine (Pinus taeda L.). A thorough understanding of below-ground carbon dynamics is necessary for the estimation of net ecosystem productivity and the carbon storage potential of these managed systems. In April 2004, we began monitoring total soil CO2 efflux (EC), heterotrophic (RH), and root respiration (RR) in response to fertilization with diammonium phosphate (DAP). Respiratory components were measured prior to fertilization, weekly following fertilization, and bi-weekly after respiratory components stabilized using a dynamic closed chamber and an infrared gas analyzer. We found that EC differed significantly (P<0.0001) between fertilized and unfertilized plots, but the direction was dependent on date. In the early period of the study, fertilized plot values were lower than control plots. However, by the latter periods fertilized plot values returned to control levels except for one sampling date in March 2005 when fertilized plot values were greater then control plots. Heterotrophic respiration was consistently and significantly (P=0.0002) lower in fertilized plots. Root respiration was significantly (P=0.0597) increased in fertilized plots when analyzed over the study and showed a 20% increase due to fertilization. We concluded that an increase in RR and possibly root biomass was enough to balance the decrease in RH leading to no difference in EC later in the growing season. We performed a pair of greenhouse studies to observe the effects of fertilization in the form of diammonium phosphate (DAP) on RR. The objectives were to determine how nutrient additions initially affect RR in one-year-old loblolly pine seedlings. Secondly, we wanted to determine if Captan [N-(trichloromethylthio) cyclohex-4-ene-1, 2-dicarboximide], a mild fungicide, could be used to reduce or eliminate ecto-mycorrhizae upon visual inspection. Both studies showed that initially, at a high rate (100 ppm N and 49 ppm P) of fertilization, RR was significantly (P<0.10) increased relative to seedlings that did not receive fertilization. This increase was only temporary with rates returning to, or decreasing below, control levels by the end of the study. No consistent trend was found between low (25 ppm N and 13 ppm P) and moderate (50 ppm N and 25 ppm P) rates of fertilization. Captan was shown to generally have no affect on RR. Captan and fertilization both showed (visual inspection) a decrease in fine-roots and mycorrhizae, which could explain the reduction in respiration rates observed in these treatments by the end of the studies. / Master of Science

Afin de mieux comprendre et évaluer l'impact du changement climatique sur les stocks de carbone organique du sol (COS) et les flux de carbone, et en particulier la respiration hétérotrophe du sol (RS), il est nécessaire d'étudier la sensibilité de la RS à la température. Plusieurs études sur les facteurs contrôlant la dépendance de la RS à la température ont été faites antérieurement et montrent que la sensibilité de la RS à la température diminue avec la température. Ces études ont suggéré que cette diminution de sensibilité de la RS à la température était liée à la modification de la disponibilité en substrat. D'autres études ont suggéré que cette diminution est liée à l'adaptation de la communauté microbienne aux températures élevées. La sensibilité de la RS à la température est plus particulièrement critique dans les régions semi-arides, comme le Nord Ouest de la Tunisie où le stock de COS est faible. Il est nécessaire de connaître l'effet de la disponibilité en substrat sur la sensibilité de la RS à la température. Dans cette étude, les échantillons de sol ont été incubés pendant 28 jours après une période de 28 jours de pré-incubation. Les pré-incubations et incubations ont été réalisées à 20, 30, 40 et 50°C. Pour tester l'effet de la disponibilité en substrat sur la sensibilité de la RS à la température, du glucose a été ajouté au sol au début de la période d'incubation. L'analyse des résultats a montré que les températures élevées de pré-incubation réduisent la sensibilité de la RS à la température d'incubation. L'addition de glucose réduit l'effet de températures élevées de pré-incubation sur la réponse de la RS. Il apparaît ainsi que la diminution de la sensibilité de la RS après un mois de pré-incubation aux fortes températures est due à la diminution de la disponibilité en substrat. Elle semble aussi liée à une diminution de la biomasse microbienne. Puisque, le sol utilisé pour cette étude est un Cambi sol Calco-magnésimorphe, une deuxième expérimentation a été réalisée afin de déterminer la part de CO2 provenant des carbonates et la part de CO2 provenant du COS. Cette étude a été réalisée par des mesures des signatures isotopiques (δ13C) du COS, des carbonates et du CO2 émis. Après 28 jours d'incubation, 23±9% de CO2 provient des carbonates. Cette contribution est faible par rapport à la teneur initiale élevée de C-CaCO3 dans le sol (4,3%) et est indépendante de la température d'incubation. Cette étude a montré que la réduction de la sensibilité de la RS aux fortes températures est surtout liée à une réduction de la disponibilité en substrat organique et de la biomasse microbienne aux fortes températures. / To better understand and assess the impact of climate change on the stocks of soil organic carbon (SOC) and carbon fluxes, and particularly heterotrophic soil respiration (SR), it is necessary to study the sensitivity of SR to temperature. Several studies have been achieved to improve the understanding of factors controlling the temperature dependence of SR and showed that the temperature sensitivity of SR decreases with temperature. These studies suggested that this decrease in temperature sensitivity of SR was related to change in substrate availability. Other studies presented microbial adaptation to warmed conditions. The temperature sensitivity of SR is especially critical in semi-arid regions, such as North West Tunisia, where the SOC stock is low. It is necessary to know the influence of substrate availability on the sensitivity of SR to temperature. In this study, soil samples were incubated for 28 days after a 28-day pre-incubation per iod. Pre-incubation and incubation were carried out at 20, 30, 40 and 50°C. To test the substrate availability effect on the temperature sensitivity of SR, glucose was added to soil at the beginning of the incubation period. Results showed that the highest pre-incubation temperature reduced the temperature sensitivity of SR during the subsequent incubation period. Glucose addition reduced the effect of high pre-incubation temperature on SR response. Thus, it appears that the observed decrease in SR sensitivity to temperature after one month pre-incubation at high temperature was due to a reduce in substrate availability and to a decrease in microbial biomass. Since the soil used in this study is a Calcari-Leptic Cambisol, a second experiment was also performed to determine the amount of CO2 from carbonates and the amount of CO2 from SOC. This study was carried out by measurements of the isotopic signatures (δ13C) of SOC, carbonates and emitted CO2. After 28 days of inc ubation, 23±9% of CO2 came from carbonates. This contribution was low compared to the high initial C-CaCO3 content in soil (4.3%), and it was independent to the incubation temperature. This study showed that reduce in the sensitivity of SR to high temperatures was probably due to a reduction in the substrate availability and to a decrease in microbial biomass.

Flow cytometry (FCM) is a well established technique used for enumeration and characterization of marine biological particles, which fulfills the scientific demands of rapid cell counting automation. FCM allows the discrimination of pico- and nanoplankton populations regarding its abundance, cell size, and pigment content both by natural or induced fluorescence. The cyanobacterium Prochlorococcus is widespread in the euphotic zone of the tropical and subtropical oceans, and is considered the smallest and most abundant photosynthetic organism in the planet. Synechococcus, other important cyanobacterium genus present in the picoplankton, is highly diverse and is widely distributed in marine ecosystems from cold and mesotrophic, to warm, open ocean oligotrophic waters. Photosynthetic pico- and nanoeukaryotes display a range of physiologies and life strategies. Although its abundance is generally lower, the larger cell size leads to a significant contribution to the epipelagic community biomass. In this thesis I aimed to investigate the abundance and carbon biomass distribution of heterotrophic bacteria, Prochlorococcus, Synechococcus, autotrophic pico- and nanoeukaryotes in the Southwest Atlantic Ocean off Brazil, their relation to the different water masses and the influence of hydrodynamic (South Atlantic Central Water intrusion) and biological processes (Trichodesmium spp. and Mesodinium rubrum blooms) on such distributions. / A citometria de fluxo (FCM, sigla em inglês) é uma técnica bem estabelecida, usada para enumeração e caracterização de partículas biológicas marinhas, a qual supre a demanda científica por automação rápida de contagem de células. A FCM permite a discriminação de populações pico- e nanoplanctônicas no que concerne à sua abundância, tamanho de célula e pigmentação, tanto por fluorescência natural ou induzida. A cianobactéria Prochlorococcus é amplamente disseminada pela zona eufótica dos oceanos tropicais e subtropicais, sendo considerada o menor e mais abundante organismo fotossintético do planeta. Synechococcus, outro importante gênero de cianobactérias presente no picoplâncton, é diverso e amplamente distribuído em ecossistemas marinhos, de águas frias e mesotróficas à regiões oceânicas quentes e de águas oligotróficas. Pico- e nanoeucariotos fotossintéticos apresentam uma grande variedade de fisiologias e estratégias de vida. Embora a abundância destes grupos seja geralmente menor, o maior tamanho de suas células resulta numa contribuição significante dos mesmos para a biomassa da comunidade epipelágica. Na presente tese eu objetivei investigar a abundância e a distribuição da biomassa de carbono de bactérias heterotróficas, Prochlorococcus, Synechococcus, pico- e nanoeucariotos autotróficos no sudoeste do Oceano Atlântico ao largo do Brasil, sua relação com as diferentes massas d\'água e a influência de processos hidrodinâmicos (como a intrusão da Água Central do Atlântico Sul) e biológicos (florações de Trichodesmium spp. e Mesodinium rubrum) em tais distribuições.

Submitted by (edna.saturno@ufrpe.br) on 2017-02-13T14:16:14Z No. of bitstreams: 1 Freddy Vogeley de Carvalho.pdf: 463787 bytes, checksum: b80fbb83db5f99c4b734647939d38607 (MD5) / Made available in DSpace on 2017-02-13T14:16:14Z (GMT). No. of bitstreams: 1 Freddy Vogeley de Carvalho.pdf: 463787 bytes, checksum: b80fbb83db5f99c4b734647939d38607 (MD5) Previous issue date: 2010-02-12 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / In the present study compared the growth performance of Litopenaeus vannamei and Farfantepenaeus subtilis in nursery heterotrophic system. The experimental design consisted of two treatments (L. vannamei and F. subtilis) with four replications. The cultivation was carried out during 52 days in tanks like "raceway" (0.9m2 and 300L), with initial stocking density of 222 shrimp/m2 and initial weight of 0.024g and 0.034g for L. vannamei and F. subtilis. The animals were fed a commercial ration (35% crude protein). The formation of heterotrophic environment was induced by adding organic fertilizer (molasses) to adjust the relationship Carbon:Nitrogen (20:1). The physical and chemical parameters of water quality showed no significant differences between treatments. The assessment of growth performance showed significant differences between the species with the highest growth (0.31g/week), higher mean final weight (2.31g) and better-feed conversion factor (0.70) for L. vannamei, compared to F. subtilis with a growth of 0.12g/week, final average weight of 0.88g and feed conversion factor of 1.09. However, F. subtilis had a higher survival (83.50%) than the L. vannamei (63.25%). Despite the best performance obtained by L. vannamei, the results indicate the potential for nursery F. subtilis in heterotrophic systems. / No presente estudo foi comparado o desempenho zootécnico de Litopenaeus vannamei e Farfantepenaeus subtilis em berçário de sistema heterotrófico. O desenho experimental foi constituído por dois tratamentos (L. vannamei e F. subtilis), com quatro repetições. O cultivo foi realizado durante 52 dias em tanques tipo “raceway” (0,9m2 e 300L), com densidade inicial de estocagem de 222 camarões/m2 e peso inicial de 0,024g e 0,034g para L. vannamei e F. subtilis, respectivamente. Os animais foram alimentados com ração comercial (35% proteína bruta). A formação do ambiente heterotrófico foi induzida através da adição de fertilizante orgânico (melaço) para ajuste da relação Carbono:Nitrogênio (20:1). As variáveis físicas e químicas de qualidade da água não apresentaram diferenças significativas entre os tratamentos. A avaliação do desempenho zootécnico apresentou diferenças significativas entre as espécies, com maior crescimento (0,31g/semana), maior peso médio final (2,31g) e melhor fator de conversão alimentar (0,70) para o L. vannamei, quando comparado ao F. subtilis que apresentou crescimento de 0,12g/semana, peso médio final de 0,88g e fator de conversão alimentar de 1,09. Entretanto, o F. subtilis obteve maior sobrevivência (83,50%) que o L. vannamei (63,25%). Apesar do melhor desempenho zootécnico obtido pelo L. vannamei, os resultados indicam potencial para o berçário do F. subtilis em sistemas heterotróficos.

Flow cytometry (FCM) is a well established technique used for enumeration and characterization of marine biological particles, which fulfills the scientific demands of rapid cell counting automation. FCM allows the discrimination of pico- and nanoplankton populations regarding its abundance, cell size, and pigment content both by natural or induced fluorescence. The cyanobacterium Prochlorococcus is widespread in the euphotic zone of the tropical and subtropical oceans, and is considered the smallest and most abundant photosynthetic organism in the planet. Synechococcus, other important cyanobacterium genus present in the picoplankton, is highly diverse and is widely distributed in marine ecosystems from cold and mesotrophic, to warm, open ocean oligotrophic waters. Photosynthetic pico- and nanoeukaryotes display a range of physiologies and life strategies. Although its abundance is generally lower, the larger cell size leads to a significant contribution to the epipelagic community biomass. In this thesis I aimed to investigate the abundance and carbon biomass distribution of heterotrophic bacteria, Prochlorococcus, Synechococcus, autotrophic pico- and nanoeukaryotes in the Southwest Atlantic Ocean off Brazil, their relation to the different water masses and the influence of hydrodynamic (South Atlantic Central Water intrusion) and biological processes (Trichodesmium spp. and Mesodinium rubrum blooms) on such distributions. / A citometria de fluxo (FCM, sigla em inglês) é uma técnica bem estabelecida, usada para enumeração e caracterização de partículas biológicas marinhas, a qual supre a demanda científica por automação rápida de contagem de células. A FCM permite a discriminação de populações pico- e nanoplanctônicas no que concerne à sua abundância, tamanho de célula e pigmentação, tanto por fluorescência natural ou induzida. A cianobactéria Prochlorococcus é amplamente disseminada pela zona eufótica dos oceanos tropicais e subtropicais, sendo considerada o menor e mais abundante organismo fotossintético do planeta. Synechococcus, outro importante gênero de cianobactérias presente no picoplâncton, é diverso e amplamente distribuído em ecossistemas marinhos, de águas frias e mesotróficas à regiões oceânicas quentes e de águas oligotróficas. Pico- e nanoeucariotos fotossintéticos apresentam uma grande variedade de fisiologias e estratégias de vida. Embora a abundância destes grupos seja geralmente menor, o maior tamanho de suas células resulta numa contribuição significante dos mesmos para a biomassa da comunidade epipelágica. Na presente tese eu objetivei investigar a abundância e a distribuição da biomassa de carbono de bactérias heterotróficas, Prochlorococcus, Synechococcus, pico- e nanoeucariotos autotróficos no sudoeste do Oceano Atlântico ao largo do Brasil, sua relação com as diferentes massas d\'água e a influência de processos hidrodinâmicos (como a intrusão da Água Central do Atlântico Sul) e biológicos (florações de Trichodesmium spp. e Mesodinium rubrum) em tais distribuições.