SILVA, Valéria Maria Araújo. Facilitação pode incrementar a capacidade de adaptação de actinobactérias e rizóbios "in vitro". 2016. 89 f. Dissertação (Mestrado em ecologia e recursos naturais)- Universidade Federal do Ceará, Fortaleza-CE, 2016. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-07-28T16:41:56Z No. of bitstreams: 1 2016_tese_vmasilva.pdf: 1606701 bytes, checksum: fe47fb5ee02d36bddccb38ea078f8a37 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-08-02T14:41:59Z (GMT) No. of bitstreams: 1 2016_tese_vmasilva.pdf: 1606701 bytes, checksum: fe47fb5ee02d36bddccb38ea078f8a37 (MD5) / Made available in DSpace on 2016-08-02T14:41:59Z (GMT). No. of bitstreams: 1 2016_tese_vmasilva.pdf: 1606701 bytes, checksum: fe47fb5ee02d36bddccb38ea078f8a37 (MD5) Previous issue date: 2016 / The natural environment is marked by an intricate network of biotic interactions that shape the structure of ecological communities. The presence of positive ecological interactions between microbial populations in soils semiarid regions, has great importance in structuring the local soil microbiota. In this work, actinomycetes strains of rhizobia and coming from rhizosphere of Park National Ubajara-CE, were evaluated for the ability to grow through cooperative metabolic mechanisms. Of the 27 evaluated actinomycetes, 22 showed compatibility with rhizobia. The strains UB-05, UB-07, UB-08, UB-11 and UB-21 stood out in facilitating tests for amylase and cellulase. The metabolic activity of actinomycetes helped the development of rhizobia strains. / O ambiente natural é marcado por uma intrincada rede de interações bióticas que moldam a estrutura das comunidades ecológicas. A presença de interações ecológicas positivas entre populações microbianas em solos de regiões semiáridas, possui grande relevância na estruturação da microbiota do solo local. Neste trabalho, cepas de actinobactérias e rizóbios oriundas de solo rizosférico do Parque Nacional de Ubajara-CE, foram avaliadas quanto à capacidade de crescerem através de mecanismos metabólicos cooperativos. Das 27 actinobactérias avaliadas, 22 apresentaram compatibilidade com rizóbios. As cepas UB-05, UB-07, UB-08, UB-11 e UB-21, destacaram-se nos ensaios de facilitação para amilase e celulase. A atividade metabólica de actinobactérias auxiliou o desenvolvimento das cepas de rizóbios.

Secretaria da EducaÃÃo do Cearà / The natural environment is marked by an intricate network of biotic interactions that shape the structure of ecological communities. The presence of positive ecological interactions between microbial populations in soils semiarid regions, has great importance in structuring the local soil microbiota. In this work, actinomycetes strains of rhizobia and coming from rhizosphere of Park National Ubajara-CE, were evaluated for the ability to grow through cooperative metabolic mechanisms. Of the 27 evaluated actinomycetes, 22 showed compatibility with rhizobia. The strains UB-05, UB-07, UB-08, UB-11 and UB-21 stood out in facilitating tests for amylase and cellulase. The metabolic activity of actinomycetes helped the development of rhizobia strains / O ambiente natural à marcado por uma intrincada rede de interaÃÃes biÃticas que moldam a estrutura das comunidades ecolÃgicas. A presenÃa de interaÃÃes ecolÃgicas positivas entre populaÃÃes microbianas em solos de regiÃes semiÃridas, possui grande relevÃncia na estruturaÃÃo da microbiota do solo local. Neste trabalho, cepas de actinobactÃrias e rizÃbios oriundas de solo rizosfÃrico do Parque Nacional de Ubajara-CE, foram avaliadas quanto à capacidade de crescerem atravÃs de mecanismos metabÃlicos cooperativos. Das 27 actinobactÃrias avaliadas, 22 apresentaram compatibilidade com rizÃbios. As cepas UB-05, UB-07, UB-08, UB-11 e UB-21, destacaram-se nos ensaios de facilitaÃÃo para amilase e celulase. A atividade metabÃlica de actinobactÃrias auxiliou o desenvolvimento das cepas de rizÃbios.

Many omics-level studies have been undertaken on Aiptasia, however, our understanding of the genes and processes associated with symbiosis regulation and maintenance is still limited. To gain deeper insights into the molecular processes underlying this association, we investigated this relationship using multipronged approaches combining next generation sequencing with metabolomics and immunohistochemistry. We identified 731 high-confident symbiosis-associated genes using meta-analysis. Coupled with metabolomic profiling, we exposed that symbiont-derived carbon enables host recycling of ammonium into nonessential amino acids, which may serve as a regulatory mechanism to control symbiont growth through a carbon-dependent negative feedback of nitrogen availability to the symbiont. We then characterized two symbiosis-associated ammonium transporters (AMTs). Both of the proteins exhibit gastrodermis-specific localization in symbiotic anemones. Their tissuespecific localization consistent with the higher ammonium assimilation rate in gastrodermis of symbiotic Aiptasia as shown by 15N labeling and nanoscale secondary ion mass spectrometry (NanoSIMS). Inspired by the tissue-specific localization of AMTs, we investigated spatial expression of genes in Aiptasia. Our results suggested that symbiosis with Symbiodiniaceae is the main driver for transcriptional changes in Aiptasia. We focused on the phagosome-associated genes and identified several key factors involved in phagocytosis and the formation of symbiosome. Our study provided the first insights into the tissue specific complexity of gene expression in Aiptasia. To investigate symbiosis-induced response in symbiont and to find further evidence for the hypotheses generated from our host-focused analyses, we explored the growth and gene expression changes of Symbiodiniaceae in response to the limitations of three essential nutrients: nitrogen, phosphate, and iron, respectively. Comparisons of the expression patterns of in hospite Symbiodiniaceae to these nutrient limiting conditions showed a strong and significant correlation of gene expression profiles to the nitrogen-limited culture condition. This confirmed the nitrogen-limited growing condition of Symbiodiniaceae in hospite, and further supported our hypothesis that the host limits the availability of nitrogen, possibly to regulate symbiont cell density. In summary, we investigated different molecular aspects of symbiosis from both the host’s and symbiont’s perspective. This dissertation provides novel insights into the function of nitrogen, and the potential underlying molecular mechanisms, in the metabolic interactions between Aiptasia and Symbiodiniaceae.

L'exposition aux mélanges de contaminants (environnementaux, alimentaires ou thérapeutiques) soulève de nombreuses interrogations et inquiétudes vis-à-vis des probabilités d'interactions toxicocinétiques et toxicodynamiques. Une telle coexposition peut influencer le mode d’action des composants du cocktail et donc de leur toxicité, suite à un accroissement de leurs concentrations internes. Le bisphénol A (4 dihydroxy-2,2-diphenylpropane) est un contaminant chimique répandu de manière ubiquitaire dans notre environnement, largement utilisé dans la fabrication des plastiques avec l’un des plus grands volumes de production à l’échelle mondiale. Il est un perturbateur endocrinien par excellence de type œstrogèno-mimétique. Cette molécule est biotransformée en métabolites non toxiques par un processus de glucuronidation. L'exposition concomitante à plusieurs xénobiotiques peut induire à la baisse le taux de glucuronidation du polluant chimique d'intérêt, entre autres la co-exposition avec des médicaments. Puisque la consommation de produits thérapeutiques est un phénomène grandissant dans la population, la possibilité d’une exposition simultanée est d’autant plus grande et forte. Sachant que l'inhibition métabolique est le mécanisme d'interaction le plus plausible pouvant aboutir à une hausse des niveaux internes ainsi qu’à une modulation de la toxicité prévue, la présente étude visait d'abord à confirmer et caractériser ce type d'interactions métaboliques entre le bisphénol A et le naproxène, qui est un anti-inflammatoire non stéroïdiennes (AINS), sur l'ensemble d'un organe intact en utilisant le système de foie de rat isolé et perfusé (IPRL). Elle visait ensuite à déterminer la cinétique enzymatique de chacune de ces deux substances, seule puis en mélange binaire. Dans un second temps, nous avons évalué aussi l’influence de la présence d'albumine sur la cinétique métabolique et le comportement de ces deux substances étudiées en suivant le même modèle de perfusion in vivo au niveau du foie de rat. Les constantes métaboliques ont été déterminées par régression non linéaire. Les métabolismes du BPA et du NAP seuls ont montré une cinétique saturable avec une vélocité maximale (Vmax) de 8.9 nmol/min/ mg prot de foie et une constante d'affinité de l'enzyme pour le substrat (Km) de 51.6 μM pour le BPA et de 3 nmol/min/mg prot de foie et 149.2 μM pour le NAP. L'analyse des expositions combinées suggère une inhibition compétitive partielle du métabolisme du BPA par le NAP avec une valeur de Ki estimée à 0.3542 μM. Les résultats obtenus montrent que l’analyse de risque pour les polluants environnementaux doit donc prendre en considération la consommation des produits pharmaceutiques comme facteur pouvant accroitre le niveau interne lors d’une exposition donnée. Ces données in vivo sur les interactions métaboliques pourraient être intégrées dans un modèle pharmacocinétique à base physiologique (PBPK) pour prédire les conséquences toxicococinétique (TK) de l'exposition d'un individu à ces mélanges chimiques. / Exposure to mixtures of contaminants (environmental, alimentary or therapeutic) poses many questions and concerns about the probability of toxicokinetic and toxicodynamic interactions. Such co-exposure may influence the mode of action of the cocktail of components and therefore their toxicity, as a result of an increase of their internal concentrations. Bisphenol A (2,2-dihydroxy-4 diphenylpropane) is a ubiquitous chemical contaminant in our environment, widely used in the manufacture of plastics, with one of the largest production volumes globally. It is an endocrine disruptor of oestrogen-mimetic type. This molecule is metabolized into non-toxic metabolites by glucuronidation process. Several factors including the co-exposure to other xenobiotics may reduce the glucuronidation rate of the chemical pollutant of interest, such as a co-exposure with drugs. Given that the consumption of therapeutic products is a growing phenomenon in the population, simultaneous exposures of pollutants with medicinal drugs are becoming a concern for health risk assessment. Knowing that metabolic inhibition is the most plausible mechanism of interaction that could result in an increase of the internal levels and a modulation of the expected toxicity, the present study aimed at confirming and characterizing this type of metabolic interactions between bisphenol A and naproxen, which is a non-steroidal anti-inflammatory drug (NSAID), on the set of an intact organ using isolated and perfused rat liver system (IPRL). The study also served to determine the enzyme kinetics metabolism of each substance alone and in binary mixture. In a second step, we also evaluated the influence of the presence of albumin on the metabolic kinetics and behavior of the two substances studied using the same in vivo perfusion model of rat liver. Metabolic constants were determined by nonlinear regression. The metabolism of BPA and NAP alone demonstrated saturable kinetics with maximal velocity (Vmax) and affinity constant (Km) equal to 8.9 nmol/min/mg prot liver and 51.6 μM for BPA, and 3 nmol/min/mg prot liver and 149.2 μM for NAP. The analysis of combined exposures suggests a partial competitive inhibition of BPA metabolism by NAP with a Ki value estimated at 0.3542 µM. From these results, it appears that the risk assessment of environmental pollutants must therefore consider the consumption of pharmaceutical products as an important agent that can increase the internal levels for a given exposure. Accordingly, these in vivo data on metabolic interactions can be incorporated into a physiologically based pharmacokinetic models (PBPK) to predict the toxicokinetic consequences (TK) of the exposure by an individual to these chemical mixtures.

La population générale est exposée à un grand nombre de pesticides, principalement via son alimentation. Alors que ces matières actives ont reçu une autorisation de mise sur le marché individuelle, l’effet sur l’organisme humain d’un mélange de tels composés est en revanche très peu connu. Afin de contribuer à mieux appréhender cet « effet mélange », notre étude s’est attachée à étudier le métabolisme hépatique humain in vitro de deux mélanges de pesticides communément retrouvés dans l’alimentation française / General population is exposed to several pesticides, mainly via the diet. Whereas these active ingredients obtained their marketing authorization individually, the available data regarding their impact as a mixture on the human body are scarce. In order to help better understanding this “mixture effect”, we aimed at studying the human hepatic in vitro metabolism of two pesticides mixtures commonly found in the diet