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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The sugarcane crop is the major agricultural activity in the coastal tableland ecosystems in Alagoas State, exerting a remarkable influence in the economical and social aspects. This activity has been continuously and intensively conducted for several years and the necessity of environmental conservation and preservation of the several natural resources in this agroecosystem makes the adoption of sustainable practices mandatory for this activity. Among these resources, soil assumes a primary role and the evaluation of its quality through the observation and analysis of physical, chemical and biological indicators that are sensitive to management practices and robust to seasonal variations may be used to define this agroecosystem sustainability. Therefore, the objective of this work was to select indicator variables of soil conservation that fulfill the criteria of sensitivity to management and robustness to seasonal variability. For this, ten composite soil samples (0-10 cm) were taken from areas grown with organic sugarcane (OS), unburned sugarcane (US), and burned sugarcane (BS), as well as from an adjacent native forest (NF) used as a reference of the undisturbed condition. Sampling was carried out at two dates: July 2005 (rainy season) and March 2006 (dry season). The following variables were analyzed: microbial biomass-C (MBC), total organic C (TOC), soil enzyme activity by the fluorescein diacetate (FDA) hydrolysis technique, the weighed mean diameter of water stable soil aggregates (WSA), and the percent of macroaggregates (PMA). MBC and TOC were higher in NF than in the cultivated areas. No differences were observed in these C pools between the three sugarcane management treatments. The response of FDA to the crop management was dependent on the sampling time. In the rainy period, the activity followed the order: NF > OS > US > BS; in the dry season, only NF differed from the other treatments. Irrespective to the sampling time, WSA and PMA decreased according with the order NF > OS = US > BS. The variables WSA and PMA were quite sensitive to discriminate between sugarcane managements, and, at the same time, robust to seasonal variations. As an additional study, we analyzed the distribution of enzyme activity (fluorescein diacetate hydrolisys) across different diameter classes of water stable aggregates in soil samples from NF and US in order to verify the relationship between these variables. It was suggested the occurrence of two pools of fluorescein diacetate catalyzing enzymes in soils from NF and US. A non-monotonic relationship was observed between aggregate diameter and FDA. For both soils, intermediate aggregate size classes (0.25 to 1.00 mm) had lower FDA values than the classes bellow and above this diameter range. / No ecossistema dos tabuleiros costeiros do Estado de Alagoas a cultura da cana-deaçúcar destaca-se como a principal atividade agrícola exercendo influência nos aspectos econômico e ambiental da região. Esta atividade tem sido praticada de forma contínua e intensa ao longo do tempo e a necessidade de conservação ambiental e preservação dos diversos recursos presentes neste agroecossistema têm exigido que a sua utilização seja sustentável. Entre os recursos existentes, o solo assume papel importante e a avaliação da sua qualidade através da observação e análise dos indicadores físicos, químicos e biológicos do solo que sejam sensíveis às práticas de manejo e resistentes a variações sazonais podem ser utilizadas para definir a sustentabilidade deste agroecossistema. Assim o objetivo deste trabalho foi selecionar variáveis indicadoras de conservação do solo que atendam aos critérios de sensibilidade ao manejo e robustez as variações sazonais. Para tanto, dez amostras compostas de solo (0-10 cm) foram coletadas de áreas sob cana-orgânica (CO), cana-crua (CC), e cana-queimada (CQ), bem como de uma mata nativa (MN) adjacente usada como referência da condição não-manejada. A amostragem foi realizada em duas datas: julho de 2005 (chuva) e março de 2006 (seca). Avaliaram-se o C da biomassa microbiana (CBM), o C orgânico total (COT), a atividade enzimática do solo pela taxa de hidrólise do diacetato de fluoresceína (FDA), o diâmetro médio ponderado de agregados estáveis em água (DMA) e a percentagem de macroagregados (PMA). O CBM e COT foram mais elevados na MN que nas áreas cultivadas, não diferindo, no entanto entre os três sistemas de manejo da cana-deaçúcar. A resposta de FDA aos manejos foi dependente da época de amostragem. No período chuvoso, os valores desta variável seguiram a ordem: MN > CO > CC > CQ; na época seca, apenas MN diferiu dos demais tratamentos. Independentemente da data de coleta, DMA e PMA decresceram na ordem MN > CO = CC > CQ. As variáveis DMA e PMA apresentaram sensibilidade satisfatória para diferenciar manejos de cana-deaçúcar e robustez à data de coleta. Após estes estudos foi analisada a distribuição da atividade enzimática medida pela hidrólise de FDA nas diferentes classes de agregados estáveis em água em amostras de solos de MN e cultivo de cana crua CC. Buscou-se com esta análise, averiguar o grau de associação entre o estado de agregação do solo e a atividade enzimática. Foi evidenciada a existência de uma compartimentalização diferenciada dos reservatórios de enzimas catalisadoras da hidrólise do FDA entre CC e MN. Observou-se uma relação não-monotônica entre estas duas variáveis. Para ambos os solos, classes de agregados intermediárias (0,25 a 1,00 mm) apresentaram menores valores de FDA que as classes abaixo e acima desta faixa de diâmetros.

Background: Cancer is a common disease, and the choice of treatment becomes more difficult over time due to chemotherapy resistant in cancer cells. To improve the in vitroassay and the individual cancer treatment, a luminescence-based endpoint assay, CellTiter Glo 2.0 was compared with the currently in use fluorescence endpoint assay, fluorometric microculture cytotoxic assay.  Aim: The aim of this study was to validate and compare the CellTiter Glo 2.0 assay with awell-established method (FMCA) and MTT [3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2Htetrazolium bromide] assay. Moreover, investigate whether the generated data can be used as a reference database for validation of patient samples in the future.   Materials and methods: The validation was performed on peripheral blood mononuclear cells from different healthy donors and two cell lines (HCT116-wt and HT-29) of colorectal cancer carcinoma were ordered frozen from American Type Culture Collection. Analysis was also done in solid samples (ovarian and kidney cancer cells). To get as correct evaluation as possible all materials were analyzed in parallel between the two methods.  Results and conclusion: A clear trend was observed when using CellTiter Glo 2.0 assay,post FMCA directly on tumor cells. This setup, makes it possible to collect reference data in the future. In addition, a high spread of the survival index data was noted between the two methods. The reason is still unknown but could be due to the low number of tested tumor cells, therefore more tumor cells need to be tested in future studies

Les processus microbiens ont une importance déterminante dans la dynamique des processus réactifs dans les eaux souterraines. La compréhension de la variabilité spatiale et temporelle de ces phénomènes, et le développement de méthodes expérimentales de terrain, ouvrent de nouveaux champs de recherches et d'applications allant de la remédiation des aquifères contaminés à la compréhension des grands cycles biogéochimiques naturels. Dans le premier volet de cette thèse nous présentons des observations de terrain permettant de comprendre le rôle des fractures sur la formation d'un ''hotspot'' d'activité microbienne en profondeur. Du fait de leur forte réactivité, ces ''hotspots'' peuvent dominer la dynamique biogéochimique des milieux souterrains, malgré leur faible extension spatiale. Nous avons ainsi analyser les conditions de formation d'un tapis microbien par des bactéries ferro-oxydantes à 60 mètres de profondeur sur l'observatoire de Ploemeur (réseau H+) alors que ces phénomènes ont été observé jusqu'à présent en surface. Les résultats de cette étude montrent que des circulations hétérogènes, liées à la structure des milieux fracturés, créent des zones mélanges entre des eaux riches en fer et des eaux oxygénées, à l'origine de ce hotspot microbien. Le deuxième volet de ce travail de thèse a été consacré au développement d'une méthode innovante pour la mesure en continu de l'activité microbienne dans les eaux souterraines. La méthode est basée sur l'utilisation de la Fluoréscéine DiAcétate (FDA) dont le produit de réaction peut être mesuré en continu par un fluorimètre de terrain. Après avoir testé et validé les protocoles sur des solutions enzymatiques et des eaux naturelles en laboratoire, nous avons mis en œuvre cette technique sur le terrain au cours d'expériences de traçages réactifs. Un modèle cinétique nous a permis d'approcher les résultats obtenus en laboratoire, et de comparer nos résultats de terrain aux calibrations effectuées in vitro. Cette méthode ouvre ainsi de nouvelles perspectives pour la caractérisation des processus biogéochimiques sur le terrain. / Microbial processes play a key role in controlling biogeochemical reactivity in groundwater. The understanding of the spatial and temporal variability of these phenomena and the development of novel experimental field methods, has opened new fields of research and applications, ranging from groundwater remediation to understanding of global biochemical cycles. In the first part of this thesis, we present field observations providing new insights on the role of fractures in the formation of a hotspot of microbial activity. Because of their large reactivity, these hotspots can dominate the biogeochemical dynamics of subsurface systems, despite their small spatial extent. We have thus analyzed the conditions for the formation of a microbial mat composed of iron-oxidizing bacteria at 60 meters depth in the Ploemeur fractured rock observatory (H+ network) while these phenomena are usually observed at the surface. These results show that heterogeneous flowpaths, linked to the structure of fractured media, create mixing zones between iron rich water and oxygen rich water, at the origin of the microbial hotspot. The second part of this work was devoted to the development of a novel method for a continuous measurement of microbial activity in groundwater. The method is based on the use of Fluorescein DiAcetate (FDA) whose product of reaction can be measured continuously by a field fluorimeter. After testing and validating protocols in the lab on enzymatic solutions and natural water, we have implemented this technic in the field in reactive tracer test experiments. A kinetic model allowed us to interpret the lab results, and to compare them to the field kinetics. This method thus opens new perspectives for the characterization of biogeochemical processes in the field.