A Araucaria angustifolia, conhecida como Pinheiro-do-Paraná é uma espécie ameaçada de extinção no Brasil, está inserida num bioma rico em biodiversidade, a Mata Atlântica, e muito pouco se conhece sobre os atributos microbiológicos dessas florestas. Este trabalho teve como objetivo avaliar potenciais atributos microbiológicos do solo e algumas variáveis ambientais (atributos químicos, físicos e efeito sazonal), a fim de estabelecer bioindicadores de qualidade do solo em Florestas de Araucária. Os estudos foram conduzidos em dois parques estaduais localizados em diferentes regiões do Estado de São Paulo, no município de Campos do Jordão (Parque Estadual de Campos do Jordão - PECJ), nos municípios de Apiaí e Iporanga (Parque Estadual Turístico do Alto Ribeira - PETAR) e também no município de Barra do Chapéu. Foram escolhidas três áreas no PETAR: FN floresta nativa de araucária, ND floresta de araucária com distúrbio antropogênico e PL plantio de araucária; e quatro áreas foram selecionadas no PECJ: FN floresta nativa de araucária, ND floresta de araucária com distúrbio antropogênico, PL plantio de araucária e PF plantio de araucária com ocorrência de fogo acidental. Ao acaso, foram selecionadas quinze árvores de araucária por área e sob a copa de cada uma delas foram retiradas amostras na profundidade de 0 a 20 cm, nas épocas seca e de chuva. Foram avaliados os atributos microbiológicos: arilsulfatase (ARIL), fosfatase ácida (FOSF), -glicosidase (GLIC), desidrogenase (DESID), nitrogênio (NBM) e carbono da biomassa microbiana (CBM), Número Mais Provável (NMP) de amonificantes (AMO), nitritadores (NITRI), nitratadores (NITRA) e desnitrificantes (DESN), respiração basal (RESP), respiração induzida por substrato (RIS), quociente metabólico (qCO2) e microbiano (qMIC). Além disso, foram estabelecidos perfis de ácidos graxos ligados a ésteres de fosfolipídios (EL-PLFAs) e o perfil de capacidade de utilização de substratos de carbono (Biolog). Alguns parâmetros químicos foram avaliados (pH, H++Al3+, Al3+, Ca2+, Mg2+, K+, P, S, N, carbono orgânico total e a capacidade de troca catiônica) e físicos (teores de argila, silte e areia). Verificou-se que o PL do PETAR e PF do PECJ foram relacionados aos atributos microbiológicos ARIL, DESN, CBM e qMIC nos dois períodos. A partição da variabilidade revelou que a atividade microbiana foi influenciada mais pelas variáveis físico-quimicas do solo do que pelas áreas e épocas de coleta. Foram identificados perfis de ácidos graxos semelhantes aos aqui obtidos, PETAR e PECJ, em um trabalho anterior no PECJ, o que pode ser indicação de um padrão de EL-PLFA próprio do ecossistema de araucária. Os ácidos graxos 10Me18:0 (actinobactéria), a relação ácido graxo saturado/insaturado (sat/insat), o consumo dos substratos -metil-D-glicosídeo e o ácido 2-hidroxibenzóico foram também relacionados às áreas PL (PETAR) e PF (PECJ) nos períodos avaliados. Já os ácidos graxos 18:19c (fungo) e 16:17c (bactéria Gram-), o consumo dos substratos glicose-1-fosfato, -D-lactose e ácido -hidroxibutírico estavam relacionados às áreas impactadas dos dois parques durante os períodos de seca e chuva. Portanto, esses atributos microbiológicos são potenciais indicadores de qualidade do solo em Florestas de Araucária, no entanto, novas avaliações são necessárias para as devidas validações dos bioindicadores e monitoramento dessas áreas. / Araucaria angustifolia, also called Brazil Pine, is an endangered species in Brazil. It is part of one of the richest ecosystems in biodiversity, the Atlantic Forest, and little is known about the soil microbiological attributes in this forest. The objective was to evaluate the potential of the soil microbiological attributes and some environmental variables (physical-chemical properties and seasonal effect) to act as bioindicators of soil quality in Araucaria Forests. This study was undertaken in two different state parks in the São Paulo State, in Campos do Jordão county (Campos do Jordão State Park - PECJ), and in Apiaí and Iporanga counties (Alto Ribeira Touristic State Park - PETAR) and also in Barra do Chapéu county. Three areas were surveyed in PETAR: FN Native Araucaria forest, ND Native Araucaria forest with anthropogenic disturbance, and PL Reforested Araucaria; and four areas in PECJ: FN Native Araucaria forest, ND Native Araucaria forest with anthropogenic disturbance; PL Reforested Araucaria, and PF Araucaria reforestation submitted to an accidental fire. Fifteen Araucaria trees were selected at random in each area and the soil and roots were sampled at 0 20 cm depth, in two contrasting seasons (dry and wet). The microbiological attributes evaluated were: arylsulfatase (ARIL), acid phosphatase (FOSF), -glucosidase (GLIC), dehydrogenase (DESID), carbon (CBM) and nitrogen microbial biomass (NBM), most probable number (NMP) of bacterial ammonium oxidizers (AMO), nitrite oxidizers (NITRI), nitrate oxidizers (NITRA) and denitrifiers (DESN), basal microbial respiration (RESP), substrate induced respiration (RIS), and metabolic (qCO2) and microbial quotient (qMIC). Additionally we determined the phospholipid fatty acid profiles (ELPLFA) and the community level physiological profile (Biolog). Some chemical attributes were evaluated (pH, H++Al3+, Al3+, Ca2+, Mg2+, K+, P, S, CEC, C/N ratio, total organic carbon (COT), total nitrogen (NT), as well as some physical ones (soil texture: clay, sand and silt contents). There was a high correlation between the PL of PETAR and the PF of PECJ and the microbiological attributes ARIL, DESN, CBM and qMIC in both periods. The partitioning of the variability evidenced that microbial activity was influenced in a higher degree by the physical-chemical properties of soil than by study areas or periods. We identified similar fatty acid profiles in PETAR and PECJ and they were also similar to the ones found in a previous work in PECJ, what could indicate a specific EL-PLFA pattern in the Araucaria ecosystem. The fatty acids 10Me18:0 (actinobacteria), the saturated/unsaturated fatty acid ratio (sat/insat), the substrate utilization of -methyl-D-glucoside and 2-hydroxybenzoic acid were also related to PL (PETAR) and PF (PECJ), in both periods. The 18:19c fatty acid (fungi) and 16:17c (Gram-), the glucose-1-phosphate, -D-lactose and -hydroxybutyric acid were C-source for microbial communities of the impacted areas in both parks during the wet and dry periods. Therefore, these attributes are potential microbial indicators of soil quality in Araucaria forests, however, new evaluations are required to confirm these biomarkers and to monitor these areas.

La pression sur les quotas de pêche et l’augmentation de la production aquacole ont contribué à une substitution importante des farines et des huiles de poisson incorporées dans les aliments pour poissons carnivores, par des farines et des huiles végétales. La truite arc-en-ciel, qui est un poisson carnivore, est affectée par ce changement de régime. Ainsi un retard de croissance apparaît dès le plus jeune stade si certaines transformations et supplémentations ne sont pas apportées aux végétaux. L’objectif de ce travail a été d’évaluer, aux stades alevins et juvéniles, l’impact d’une substitution totale des huiles et farines de poisson sur le tractus digestif de la truite arc-en-ciel, et plus particulièrement sur ses capacités digestives et sur la composition de son microbiote intestinal. Le but in fine étant de déterminer si certaines enzymes de la digestion, transporteurs intestinaux, ou sous-communautés bactériennes sont impactés par le changement de régime et peuvent expliquer le retard de croissance observé. Chacun de ces facteurs ont été étudiés via une approche de métagénomique par séquençage de nouvelle génération NGS pour la caractérisation du microbiote, et via de la PCR quantitative et des mesures d’activités enzymatiques pour la comparaison des capacités digestives. Des lignées isogéniques de truites, identifiées comme divergentes dans leur réponse à l’alimentation végétale (capables d’adaptation ou réfractaires) ont permis de disposer d’un matériel biologique pertinent pour répondre à cette question. Une modification du microbiote intestinal associé à la muqueuse digestive pourrait également contribuer à la baisse de l’homéostasie intestinale. Le changement de régime conduit en effet à une équitabilité plus faible chez les truites ayant reçue un aliment végétal, ce qui reflète un changement dans la représentativité de certains OTUs. Ce changement de régime s’est également traduit par des communautés dissimilaires en moyenne à 70 %, d’après l’estimation de la β-diversité entre les communautés de truites nourries avec l’aliment marin et celles nourries avec l’aliment végétal. La sélection opérée par l’aliment a conduit à un remplacement des OTUs rencontrés au sein des Firmicutes, c’est-à-dire que différentes espèces bactériennes de Firmicutes sont rencontrées suivant le régime considéré. La comparaison de communautés bactériennes entre les différentes lignées isogéniques a montré que la sélection opérée par le génotype de l’hôte a davantage eu lieu sur le remplacement des β-Protéobactéries. Enfin, les comparaisons d’abondances en certaines espèces bactériennes particulières suggèrent que les bactéries Cetobacterium somerae, capables de synthétiser de la vitamine B12, et Shewanella, dont l’implication dans la stimulation des cellules β du pancréas endocrine a déjà été observée chez d’autres espèces, pourraient être impliquées dans la réponse métabolique des truites aux végétaux. Les modifications identifiées dans ce travail constituent des indicateurs biologiques qui pourront être mis à profit pour évaluer la réponse du tractus digestif des truites à de nouvelles formules alimentaires. / Over-fishing pressure and increasing aquaculture production led to an important substitution of fish oil and fish meal with oil and meal from plant origin in feed meant for farming fish. However this replacement has some deleterious incidence on fish. For rainbow trout, which are carnivorous fish, some growth delay often appears from the early life stages when they are fed with plant based diet. The aim of this work was to assess, at alevin and juvenile stages, the impact of a total replacement of fish meal and oil on rainbow trout gastrointestinal tract, and more particularly on the digestive capacity and the associate microbiota. The objective, in fine, being to determine if some digestive enzymes, intestinal transporters, or bacterial communities are impacted by the dietary replacement and if these biological factors can be related to the observed growth delay. Metagenomic approach using next generation sequencing was used to characterize the gut bacterial communities, while digestive capacity was assessed through quantitative PCR and enzymatic measurements in order to compare rainbow trout responses to a plant-based diet. In our investigations, rainbow trout isogenic lines that diverge in their response to this alternative diet (tolerant or rather reluctant) were adopted because they constitute a pertinent biological material for answering this question.In alevin rainbow trout, a plant-based diet led to an increase of pepsinogen, trypsinogen, and chymotrypsinogen genes which codes for proteolitic enzymes. Two main assumptions can explain this response, and their effectivness remains to investigate: wether this is a physiological response due to a lower weight of trout fed with the plant-based diet, or so it is due to an increase transcritpion of pancreatic enzymes to compensate for a reduction of protein digestibility. In the intestine, it appears that an increase transcription of IAP, SGLT1, CCK-t, and PEPT1 genes, and a decrease transcription of GLUT2 gene under a plant-based diet could reflect a disability to grow under a vegetable diet.In juvenile rainbow trout, a plant-based diet led to a decrease of lipid digestibility, and of triglycerides and total amino acid plasmatic levels. These perturbations could be explained in part by a decrease of the phosphatase alkaline activity, which suggest perturbancesof intestinal homeostasis, and by a decrease of phospholipase A2 activity. Transcriptional decrease of the triglycerides transporter MTP and of the prolidase, which is a peptidase from intestinal cell cytosol, has also been observed. Some modification of the microbiota associated to the intestinal mucosa could also contribute to the decrease of the intestinal homeostasis. The dietary replacement effectively led to reduce evenness of the bacterial communities in trout fed with a plant-based diet, which reflected a shift in the representativeness of some OTUs. Bacterial community from trout fed with a marine diet and trout fed with a plant-based diet were on average 70 % dissimilar. Dietary substitution led to the replacement of OTUs from the Firmicutes class, different bacterial species being observed according to the considered diet. The comparison of bacterial community between the isogenic lines showed that the genotype led to the replacement of β-Proteobacteria. Finally, abundance comparison suggested that Cetobacterium somerae, which is able to synthesise vitamin B12, and Shewanella, which has already been reported to stimulate pancreatic β cells, could be implicated in the trout response to vegetable.Modifications observed in this work constitute biological indicator that could be used to assess the response of the digestive tract to future feed formulations.

A Araucaria angustifolia, conhecida como Pinheiro-do-Paraná é uma espécie ameaçada de extinção no Brasil, está inserida num bioma rico em biodiversidade, a Mata Atlântica, e muito pouco se conhece sobre os atributos microbiológicos dessas florestas. Este trabalho teve como objetivo avaliar potenciais atributos microbiológicos do solo e algumas variáveis ambientais (atributos químicos, físicos e efeito sazonal), a fim de estabelecer bioindicadores de qualidade do solo em Florestas de Araucária. Os estudos foram conduzidos em dois parques estaduais localizados em diferentes regiões do Estado de São Paulo, no município de Campos do Jordão (Parque Estadual de Campos do Jordão - PECJ), nos municípios de Apiaí e Iporanga (Parque Estadual Turístico do Alto Ribeira - PETAR) e também no município de Barra do Chapéu. Foram escolhidas três áreas no PETAR: FN floresta nativa de araucária, ND floresta de araucária com distúrbio antropogênico e PL plantio de araucária; e quatro áreas foram selecionadas no PECJ: FN floresta nativa de araucária, ND floresta de araucária com distúrbio antropogênico, PL plantio de araucária e PF plantio de araucária com ocorrência de fogo acidental. Ao acaso, foram selecionadas quinze árvores de araucária por área e sob a copa de cada uma delas foram retiradas amostras na profundidade de 0 a 20 cm, nas épocas seca e de chuva. Foram avaliados os atributos microbiológicos: arilsulfatase (ARIL), fosfatase ácida (FOSF), -glicosidase (GLIC), desidrogenase (DESID), nitrogênio (NBM) e carbono da biomassa microbiana (CBM), Número Mais Provável (NMP) de amonificantes (AMO), nitritadores (NITRI), nitratadores (NITRA) e desnitrificantes (DESN), respiração basal (RESP), respiração induzida por substrato (RIS), quociente metabólico (qCO2) e microbiano (qMIC). Além disso, foram estabelecidos perfis de ácidos graxos ligados a ésteres de fosfolipídios (EL-PLFAs) e o perfil de capacidade de utilização de substratos de carbono (Biolog). Alguns parâmetros químicos foram avaliados (pH, H++Al3+, Al3+, Ca2+, Mg2+, K+, P, S, N, carbono orgânico total e a capacidade de troca catiônica) e físicos (teores de argila, silte e areia). Verificou-se que o PL do PETAR e PF do PECJ foram relacionados aos atributos microbiológicos ARIL, DESN, CBM e qMIC nos dois períodos. A partição da variabilidade revelou que a atividade microbiana foi influenciada mais pelas variáveis físico-quimicas do solo do que pelas áreas e épocas de coleta. Foram identificados perfis de ácidos graxos semelhantes aos aqui obtidos, PETAR e PECJ, em um trabalho anterior no PECJ, o que pode ser indicação de um padrão de EL-PLFA próprio do ecossistema de araucária. Os ácidos graxos 10Me18:0 (actinobactéria), a relação ácido graxo saturado/insaturado (sat/insat), o consumo dos substratos -metil-D-glicosídeo e o ácido 2-hidroxibenzóico foram também relacionados às áreas PL (PETAR) e PF (PECJ) nos períodos avaliados. Já os ácidos graxos 18:19c (fungo) e 16:17c (bactéria Gram-), o consumo dos substratos glicose-1-fosfato, -D-lactose e ácido -hidroxibutírico estavam relacionados às áreas impactadas dos dois parques durante os períodos de seca e chuva. Portanto, esses atributos microbiológicos são potenciais indicadores de qualidade do solo em Florestas de Araucária, no entanto, novas avaliações são necessárias para as devidas validações dos bioindicadores e monitoramento dessas áreas. / Araucaria angustifolia, also called Brazil Pine, is an endangered species in Brazil. It is part of one of the richest ecosystems in biodiversity, the Atlantic Forest, and little is known about the soil microbiological attributes in this forest. The objective was to evaluate the potential of the soil microbiological attributes and some environmental variables (physical-chemical properties and seasonal effect) to act as bioindicators of soil quality in Araucaria Forests. This study was undertaken in two different state parks in the São Paulo State, in Campos do Jordão county (Campos do Jordão State Park - PECJ), and in Apiaí and Iporanga counties (Alto Ribeira Touristic State Park - PETAR) and also in Barra do Chapéu county. Three areas were surveyed in PETAR: FN Native Araucaria forest, ND Native Araucaria forest with anthropogenic disturbance, and PL Reforested Araucaria; and four areas in PECJ: FN Native Araucaria forest, ND Native Araucaria forest with anthropogenic disturbance; PL Reforested Araucaria, and PF Araucaria reforestation submitted to an accidental fire. Fifteen Araucaria trees were selected at random in each area and the soil and roots were sampled at 0 20 cm depth, in two contrasting seasons (dry and wet). The microbiological attributes evaluated were: arylsulfatase (ARIL), acid phosphatase (FOSF), -glucosidase (GLIC), dehydrogenase (DESID), carbon (CBM) and nitrogen microbial biomass (NBM), most probable number (NMP) of bacterial ammonium oxidizers (AMO), nitrite oxidizers (NITRI), nitrate oxidizers (NITRA) and denitrifiers (DESN), basal microbial respiration (RESP), substrate induced respiration (RIS), and metabolic (qCO2) and microbial quotient (qMIC). Additionally we determined the phospholipid fatty acid profiles (ELPLFA) and the community level physiological profile (Biolog). Some chemical attributes were evaluated (pH, H++Al3+, Al3+, Ca2+, Mg2+, K+, P, S, CEC, C/N ratio, total organic carbon (COT), total nitrogen (NT), as well as some physical ones (soil texture: clay, sand and silt contents). There was a high correlation between the PL of PETAR and the PF of PECJ and the microbiological attributes ARIL, DESN, CBM and qMIC in both periods. The partitioning of the variability evidenced that microbial activity was influenced in a higher degree by the physical-chemical properties of soil than by study areas or periods. We identified similar fatty acid profiles in PETAR and PECJ and they were also similar to the ones found in a previous work in PECJ, what could indicate a specific EL-PLFA pattern in the Araucaria ecosystem. The fatty acids 10Me18:0 (actinobacteria), the saturated/unsaturated fatty acid ratio (sat/insat), the substrate utilization of -methyl-D-glucoside and 2-hydroxybenzoic acid were also related to PL (PETAR) and PF (PECJ), in both periods. The 18:19c fatty acid (fungi) and 16:17c (Gram-), the glucose-1-phosphate, -D-lactose and -hydroxybutyric acid were C-source for microbial communities of the impacted areas in both parks during the wet and dry periods. Therefore, these attributes are potential microbial indicators of soil quality in Araucaria forests, however, new evaluations are required to confirm these biomarkers and to monitor these areas.

Les vasières intertidales sont le siège d’une forte production primaire sous la forme d’un biofilmmicrophytobenthique qui se développe en surface à marée basse. Ce biofilm se compose principalement de diatomées et de bactéries hétérotrophes. Ces deux composantes sécrètent des substances polymériques extracellulaires (EPS) qui jouent différents rôles dans le biofilm. Le travail présenté s’est basé sur différentes échelles d’observation (in situ à 2 saisons, mésocosme en laboratoire, et échelle fine du biofilm en microscopie confocale) et a permis de mettre en évidence les interactions des diatomées et des bactéries à différents moments de la marée et du nycthémère. L’utilisation d’une nouvelle méthode d’extraction des EPS a permis d’éclaircir leur rôle en évitant les contaminations par les substances internes provoquées par les méthodes classiques. Les EPS colloïdales particulièrement riches en glucose s’associent au déplacement des diatomées, notamment lors d’un stress (sursalure, carence en nutriments) et sont préférentiellement consommées par les bactéries, après leur dégradation par les enzymes ou leur hydrolyse dans l’eau interstitielle. Les EPS liées au frustule, et plus particulièrement les sucres, inhiberaient le développement bactérien à proximité de la cellule algale. Elles sont surtout sécrétées lors de la mise en place du biofilm et pour protéger la cellule quand les conditions sont osmotiquement défavorables et leur richesse en protéine leur confère un potentiel intéressant pour les bactéries qui peuvent les utiliser comme substrat azoté en cas de carence. D’autres substances ont été plutôt sécrétées parles bactéries telles que les N-acétylglucosamines et les protéines colloïdales de bas poids moléculaire,certainement des enzymes bactériennes, ainsi que le glucose qui semble être associé au EPS colloïdaux des diatomées mais aussi aux EPS bactériens selon l’analyse en microscopie confocale en utilisant des lectines comme marqueurs d’EPS. / Microphytobenthic biofilms that develop at the intertidal mudflat surface are responsible of a substantialprimary production. These biofilms are composed of diatoms and heterotrophic bacteria, both excretingextracellular polymeric substances (EPS) that have various functions into the biofilm. The present thesis is basedon different observation scales that have highlighted the complex interactions between diatoms and bacteria fordifferent moment of the biofilm development (in situ at 2 seasons, in laboratory mesocosm and at the scale of thebiofilm at confocal microscopy). Furthermore, a new extraction method that avoids contamination with internalsubstances was used and provides new insights about the role of EPS. Colloidal EPS are excreted during thediatom migrations, notably in case of environmental stress (nutrient depletion, high salinity). They arepreferentially consumed by bacteria, after hydrolysis or enzymatic cleavage. Bound EPS are associated to abacterial inhibition, especially sugars. These EPS are mainly excreted during biofilm formation and also in caseof osmotic stress, if diatoms cannot migrate toward favorable slices. Some substances are associated to bacterialdevelopment, such as N-acetylglucosamine, low molecular weight proteins that probably consist of bacterialenzymes and also glucose that seems to be associated to colloidal EPS secreted by diatoms as well as bacterialEPS, when analyzing the biofilm with confocal microscopy by using lectins as EPS biomarkers.

El reg amb aigües residuals tractades s'ha convertit avui en dia en una necessitat, ja que permet preservar els recursos hídrics de més qualitat i constitueix una alternativa necessària per al manteniment de regadius tradicionals. En aquesta tesi s'avaluen els efectes del reg amb aigües residuals tractades d'origen domèstic sobre les característiques químiques i biològiques de diferents tipologies de sòls a l'illa de Mallorca, així com sobre el creixement dels cultius i la seva composició mineral mitjançant diferents estudis de camp i en cultius en contenidors. En general no s'han apreciat efectes negatius importants causats pel reg amb aquestes aigües excepte alguns valors puntualment més elevats de crom i plom que es relacionen amb aportacions d'aigües poc depurades realitzades en el passat. Segons els resultats obtinguts amb el reg amb aigües residuals tractades d'origen domèstic es contribueix a millorar la qualitat del sòl i s'aporten part dels nutrients que requereixen els cultius per al seu creixement. / El riego con aguas residuales tratadas se ha convertido hoy en día en una necesidad, ya que permite preservar los recursos hídricos de más calidad y constituye una alternativa necesaria para el mantenimiento de regadíos tradicionales. En esta tesis se evalúan los efectos del riego con aguas residuales tratadas de origen doméstico sobre las características químicas i biológicas de diferentes tipologías de suelos en la isla de Mallorca, así como sobre el crecimiento de los cultivos y su composición mineral mediante diferentes estudios de campo i en cultivos en contenedores. En general no se han apreciado efectos negativos importantes causados por el riego con estas aguas, excepto algunos valores puntualmente más elevados de cromo y plomo que se relaciona con aportaciones de aguas poco depuradas realizadas en el pasado. Según los resultados obtenidos con el riego con aguas residuales tratadas de origen doméstico se contribuye a mejorar la calidad del suelo y se aportan parte de los nutrientes que requieren los cultivos para su crecimiento. / Nowadays the irrigation with treated waste water has become a necessity, not only, because it allows preserving fresh water but it also constitutes an important alternative to the maintenance of traditional irrigating lands. Considering different chemical and biological characteristics of soil samples of Mallorca island (Spain), this thesis investigates the effect of irrigation with treated domestic waste water on soil properties. In addition, several studies were conducted in a series of pot experiments and land conditions to monitor the crops growth and evaluate their mineral compositions. In general, no negative effects caused by this type of irrigation have been observed, except of some high values of chromium and lead which were related to past irrigation realized with untreated effluents. According to our results, the irrigation with treated domestic wastewater contributes to improve soil quality and supplies part of the nutrient requirements of the crops.

F1FO-ATPsynthase is a key enzyme in energy metabolism of the cell. Its deficit is caused usually by mutations in two structural genes MT-ATP6 and MT-ATP8 encoded by the mitochondrial DNA or in nuclear genes ATPAF2 and TMEM70 encoding the biogenesis factors and structural gene ATP5E. Deficiency of the F1FO-ATPsynthase leads to progressive and serious phenotype affecting organs with high energy demands. The first symptoms usually occurs in neonatal age and prognosis of the disease is fatal. Mutations in these genes result in both qualitative and quantitative defects of the F1FO-ATPsynthase. The study of molecular bases of mitochondrial disorders including F1FO-ATPsynthase deficiency uses large number of biochemical and molecular-genetic methods to determine a proper diagnosis which is essential for the symptomatic therapy and genetic counselling in affected families. The aim of the diploma thesis was to characterise the F1FO-ATPsynthase deficiency in isolated mitochondria from the lines of cultured cells by the determination oligomycin- sensitive ATP-hydrolytic activity of the F1FO-ATPsynthase, enzymatic activities of the respiratory chain complexes and to analyse changes in the steady-state levels of the representative subunits and whole complex of the F1FO-ATPsynthase in comparison with controls. 3...