Made available in DSpace on 2017-07-21T18:53:07Z (GMT). No. of bitstreams: 1 Jessica Teles.pdf: 1283462 bytes, checksum: 5784cdcc4b339768e1ee02500a793d75 (MD5) Previous issue date: 2016-04-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Propionic acid is an organic acid used as a preservative in foods and seeds, and as an ingredient in thermoplastics, medicaments, perfumes, fragrances and solvents. Its production takes place by conventional chemical synthesis that uses petroleum as raw material. Finding alternatives and more sustainable ways than the use of petroleum products has been shown an interesting option to the production of organic acids. The purpose of this study was to evaluate the propionic acid production from agroindustrial effluents by fermentation, using a culture of Propionibacterium acidipropionici CCT 4843. Whey, corn steep liquor and animal feed effluent were used as substrate. The substrates composition was based on an experimental design mixture, resulting in 10 samples. The substrates were fermented in batch borosilicate glasses at a temperature of 35°C, initial pH 6.5 and 20 mL.L-1 suspension of inoculum. After choose the substrate, a 33-1 factorial design was carried out, to see how the variable hydraulic retention time, inoculum concentration and pH affected the production of propionic acid. The propionic and other organic acids were detected using a high-performance liquid chromatography (HPLC). The corn steep liquor was chosen as the best a substrate for the factorial design, because it showed the highest propionic acid yield, 0.79 g. g substrate-1,propionic acid productivity of 5.20 mg.L-1.h-1 and production of 0.40 gL-1. It was found that hydraulic retention time and initial pH influenced on the yield and propionic acid productivity, but the initial inoculum concentration did not influence the answers. The highest propionic acid yield of 0.6275 g.g- 1, was obtained with 96 hours of hydraulic retention time, inoculum of initial concentration of 20 mL.L-1 and initial pH of 6.0.The best propionic acid productivity was obtained for hydraulic retention time of 60h, initial inoculum concentration of 30 mL.L-1 and initial pH of 6.5. It was concluded that propionic acid production is possible from agroindustrial waste, but the process conditions should be optimized to make it economically viable. / O ácido propiônico é um ácido orgânico utilizado como conservante em alimentos e sementes e como ingrediente em termoplásticos, drogas, perfumes, aromas e solventes. Sua produção convencional ocorre por síntese química, utilizando-se matérias primas obtidas do petróleo. Porém, com o aumento da preocupação ambiental e procura por alternativas ao uso de produtos derivados de petróleo, a produção de ácidos orgânicos por fermentação tem se mostrado uma opção interessante. O objetivo dessa pesquisa foi avaliar a produção de ácido propiônico a partir da fermentação de efluentes agroindustriais, utilizando a cultura Propionibacterium acidipropionici CCT 4843. Os efluentes utilizados como substrato foram soro de leite, milhocina e efluente da produção de ração animal. A composição dos substratos foi determinada a partir de um planejamento experimental de misturas, resultando em 10 ensaios. Os substratos foram fermentados em batelada em reatores de vidro borosilicato à temperatura de 35°C, pH inicial de 6,5 e 20 mL.L-1 de inóculo em suspensão. Após definida a composição do substrato, foi realizado o planejamento fatorial 33-1, para verificar como as variáveis Tempo de Detenção Hidráulica (TDH), concentração de inóculo e correção inicial de pH afetam a produção de ácido propiônico. A determinação do ácido propiônico e demais ácidos orgânicos foi realizada por Cromatografia Líquida de Alta Eficiência(CLAE). No planejamento fatorial para estudo das variáveis de processo foi utilizado como substrato apenas a milhocina, pois nos ensaios do planejamento experimental de misturas foi o que possibilitou o maior rendimento, com 0,79 g ácido propiônico. g substrato-1, com uma produtividade de ácido propiônico de 5,20 mg/L.h e produção de 0,40g.L-1. No segundo planejamento foi constatado que as variáveis tempo de detenção (TDH) e o pH inicial influenciam no rendimento e na produtividade de ácido propiônico, porém a concentração inicial de inóculo não influenciou nas respostas. O melhor rendimento obtido foi de 0,6275 g.g-1, com o ensaio com TDH de 96h, concentração inicial de inóculo de 20mL.L-1 em suspensão e pH inicial 6,0 ; e o melhor valor de produtividade obtido foi para o ensaio com tempo de detenção hidráulica de 60h, concentração inicial de inóculo de 30 mL.L-1 e pH inicial de 6,5. Foi concluído que é possível a produção de ácido propiônico a partir de efluentes agroindustriais, porém as condições de processo devem ser otimizadas para torná-lo economicamente viável.

La chromatographie en phase gazeuse couplée à la spectrométrie de masse simple quadripolaire (GC-qMS) est la technique la plus utilisée dans l’exploration des acides organiques urinaires dans le cadre du diagnostic des aciduries organiques (AO). Cependant, après analyse par GC-MS de plus de 1000 échantillons d'urines recueillis auprès d'enfants atteints de troubles neurologiques, moins de 0,5% d'entre eux ont reçu un diagnostic positif. Ceci témoigne de l’urgence de rechercher de nouveaux biomarqueurs dans le domaine des troubles neurologiques associés à des erreurs innées du métabolisme (EIM).L'objectif principal de cette thèse a été de développer une nouvelle méthode d’analyse des échantillons urinaires par chromatographie bidimensionnelle couplée à la spectrométrie de masse (GCxGC-qMS) et d'évaluer son apport dans le profilage métabolique de ces échantillons. Par ailleurs, dans le cadre du diagnostic des AO, dans un objectif de simplification de la procédure de préparation de l’échantillon, nous avons évalué une autre approche d’analyse directe en temps réel couplée à la spectrométrie de masse « DART-MS » des acides organiques urinaires. La première étape de notre travail a été de développer un protocole de préparation de l’échantillon compatible avec une méthode de séparation bidimensionnelle. Puis, grâce à l’étude systématique des différentes conditions de séparation, nous avons développé une méthode par CGxCG-qMS parfaitement adaptée à la séparation des composés extraits.Après validation analytique, la méthode proposée a été ensuite évaluée par le profilage des composés organiques présents dans des échantillons urinaires prélevés chez des volontaires sains. Par rapport aux profils obtenus par une méthode par GC-qMS classique, optimisée, les profils chromatographiques obtenus par la méthode proposée présentent une sensibilité et une résolution nettement plus élevées. Ceci se traduit par la détection en GCxGC-qMS de nombreux composés supplémentaires, jusqu’à 92, selon les échantillons. Parmi les composés supplémentaires détectés et caractérisés, certains sont d’un intérêt diagnostic reconnu.Malgré les difficultés liées à l’étape de préparation de l’échantillon et au traitement des données, l’ensemble des résultats obtenus par la méthode proposée sur de nombreux échantillons urinaires a confirmé les potentialités et la nécessité d’utiliser la chromatographie bidimensionnelle dans le domaine de la découverte de candidats biomarqueurs. Il s’agit maintenant de poursuivre l’identification des composés supplémentaires détectés dans les échantillons urinaires étudiés et d’appliquer la méthode proposée à un plus grand nombre d’échantillons témoins et pathologiques afin d’essayer d’identifier des candidats biomarqueurs.Enfin, les essais préliminaires par la technique DART-MS, effectués dans un laboratoire de la Direction des Applications Militaires (DAM) du CEA, ont été axés sur la détection d’un panel d’acides organiques représentatifs des différentes anomalies rencontrées lors des AO. Malgré les effets de suppression ionique qui restent à éliminer, les résultats obtenus montrent que cette approche mérite d’être approfondie. / Among inherited metabolic diseases, organic acidemia (OA) or organic aciduria is characterized by urinary excretion of abnormal amounts or types of organic acids. OA is mostly associated with genetic conditions resulting in a specific step of amino acid catabolism dysfunction. Such alterations can produce disease states that range from mild to lethal neurological involvement. Gas chromatography coupled to mass spectrometry (GC-MS) remains the most used analytical technique for detecting specific urinary organic metabolites related to OA. However, after analyzing more than 1,000 urine samples collected from children with neurological disorders, by this technique less than 0.5% of them were positive. Thus, the discovery of new biomarker candidates for other metabolic diseases is urgently needed. The main objective of this thesis was to evaluate a new method using two-dimensional gas chromatography coupled to mass spectrometry (GCxGC-MS) for the both qualitative and quantitative metabolic profiling of children’s urine. As compared to GC-MS, GCxGC-MS shows great resolution power and high peak capacity. For this purpose, we first developed a GCxGC-MS method with an appropriate sample preparation protocol for urinary organic acids profiling. Applied to urine samples of healthy children and children with neurological disorders, the proposed method showed high sensitivity and peak capacity thus opening new possibilities for the characterization of new biomarker candidates. For instance, by this technique, we were able to detect in urines samples more than ninety additional compounds, which are not detected by a conventional GC-MS method. However, taken together the obtained results show that while the GCxGC technique has unmatched power to separate compounds in a complex mixture, the sample preparation protocol remains a limiting step for the precise quantification of the detected compounds. On another hand, in order to reduce the sample preparation step, we evaluated the direct analysis in real time (DART) method for the urinary organic acids screening. The obtained results are very promising.

Com o objetivo de estabelecer as possíveis correlações entre os ácidos orgânicos na precipitação e as alterações no uso do solo, amostras de água da chuva e aerossol foram coletadas nos Parques Estaduais de Intervales (PEI) e Morro do Diabo (PEMD), no Estado de São Paulo, cuja vegetação é formada por florestas pertencentes ao domínio Mata Atlântica. Entre julho de 2002 a julho de 2004 foram coletadas 232 amostras de água de chuva em PEI e 96 amostras em PEMD, determinando-se os teores de formato, acetato, metanosulfonato, piruvato, oxalato, maleato, valerato, citrato, butirato, tartarato, cloreto, nitrato, sulfato, sódio, amônio, potássio, cálcio e magnésio por cromatografia iônica (CI). Também foram determinadas as concentrações de carbono orgânico dissolvido (COD), carbono inorgânico dissolvido (CID) e o pH das amostras. Entre julho de 2002 a maio de 2003, amostras de aerossol inalável (PM10), subdividido nas frações fina (dp < 2,5 µm) e grossa (2,5 < dp < 10 µm), foram coletadas separadamente, totalizando 60 amostras para cada fração no PEMD e 90 amostras para cada fração no PEI, determinando-se as concentrações em massa por gravimetria e a composição química do material solúvel em água por CI. Eventos de chuva ácida ocorreram nos dois locais ao longo do período de amostragem. Os ácidos orgânicos contribuíram com até 16% da somatória iônica (µeq.L-1) no PEI e até 12% no PEMD, com predominância do ácido acético no PEI e com predominância de ácido fórmico no PEMD. A emissão direta da vegetação foi a principal fonte de ácidos orgânicos no PEI, uma vez que a razão entre o ácido fórmico e ácido acético < 1,0. No PEMD, a razão entre ácido fórmico e acético > 1,0 indicou como as principais fontes as reações secundárias de oxidação fotoquímicas no local e a emissão direta biogênica da vegetação. A contribuição dos ácidos orgânicos para a acidez da água da chuva foi de até 40% no PEI e até 30% no PEMD. Adicionalmente, as correlações significantes entre H+ e COD (p < 0,05) sugerem contribuições de compostos orgânicos para acidez da água da chuva. A influência de fontes regionais de queima de biomassa e das condições meteorológicas na composição química da atmosfera no PEMD é evidenciada pela concentração média de PM10 (22 ug.m-3); pela concentração de \"black carbon\" (40 ?g.m-3), superiores às verificadas no PEI, pela maior contribuição da fração fina do aerossol para a concentração de COD da fração solúvel em água. Por outro lado, no PEI, a fração grossa relacionada à compostos biogênicos, aerossol marinho e partículas de solo foi a maior contribuinte para o COD. Dos ácidos orgânicos detectados no aerossol solúvel em água, o ácido oxálico foi o mais abundante. / In order to establish the possible correlations among organic acids in the precipitation and land use changes, rain water and aerosol samples were collected in the State Parks of Intervales (PEI) and Morro do Diabo (PEMD), in the State of São Paulo. The vegetation there is formed by Mata Atlântica forests (Atlantic forest). Between July, 2002 and July, 2004, 232 samples of rainwater were collected in PEI and 96 samples in PEMD, for the analysis of formate, acetate, methanesulfonate, pyruvate, oxalate, malate, valerate, citrate, butyrate, tartarate, chloride, nitrate, sulfate, sodium, ammonium, potassium, calcium, magnesium for ionic chromatography (CI). The concentrations of dissolved organic carbon (COD), dissolved inorganic carbon (CID) and pH of the samples were also determined. Between July, 2002 and May, 2003, samples of inhalable aerosol (PM10), subdivided in fine fraction (dp <2.5 µm) and coarse fraction (2.5 <dp <10 µm), were collected separately, in a total of 60 samples for each fraction in PEMD and 90 samples for each fraction in PEI, submitted to the analyses of concentration in mass through gravimeter and the chemical composition of the water soluble aerosol for CI. Events of acid rain were verified along the sampling period in both places and the organic acids contributed up to 16% of the ionic sum (µeq.L-1) in PEI and up to 12% in PEMD, with predominance of acetic acid in PEI and with predominance of formic acid in PEMD. The direct emission from vegetation was the main source of organic acids in PEI, once the ratio between the formic acid and acetic acid was <1.0 and that the effects of biomass burns are not verified. In PEMD, the ratio among formic and acetic acid >1.0 indicated the secondary reactions of photo-chemical oxidation ?in situ? and biogenic direct emission from the vegetation as the main source. The contribution of the organic acids for the acidity of the rainwater was up to 40% in PEI and up to 30% in PEMD. In addition to the strong correlations between H+ and COD (p <0.05) they suggest contributions of organic compositions for the acidity of the rainwater. The influence of regional sources of biomass burning and of the meteorological conditions in the chemical composition of the atmosphere in PEMD is also observed by the medium concentration of PM10 (22 µg.m-3) and concentration of \"black carbon\" (40 ?g.m-3), both higher than the ones observed in PEI, and for the highest contribution of fine fraction of aerosol for the concentration of COD analyzed from the water soluble fraction. In PEI, the coarse fraction related to biogenic compounds, marine aerosol and soil particles were the greatest responsible agents for COD. Oxalic acid was the most abundant organic acid detected.

Por se tratar de um elemento essencial às plantas e um metal pesado ao mesmo tempo, o níquel requer atenção quanto aos aspectos da fisiologia de plantas e ambiental. Além disso, existe um intervalo estreito entre as exigências nutricionais e os teores tóxicos às plantas. Neste contexto, objetivou-se avaliar o efeito do Ni no sistema solo-planta, com foco no ciclo do N e a disponibilidade do elemento no solo, por meio de experimento em condições controladas, utilizando vasos distribuídos inteiramente ao acaso, utilizando-se esquema fatorial 2 x 5, com sete repetições cada tratamento. O primeiro fator foi constituído de duas saturações por base (50 e 70%) e o segundo de cinco doses de Ni (0; 0,1; 0,5; 1,0 e 10,0 mg dm-3 de solo). Os vasos foram preenchidos com 8 dm3 de terra e cultivados com soja [Glycine max (L.) Merrill] sucedida por girassol (Helianthus annuus L.). Os parâmetros qualitativos e quantitativos: altura de plantas (AP), diâmetro do caule (DC), número de nós (NN), estádio fenológico (EF), índice SPAD e, diâmetro do capítulo (DCap) (para girassol) foram avaliadas aos 30 e 60 dias após a emergência (d.a.e.) de cada cultivo. Plantas inteiras de soja, amostradas em quatro vasos de cada tratamento, foram coletadas no estádio R1. Na mesma ocasião foram coletadas amostras de solo da rizosfera. Em seguida, as plantas coletadas foram divididas em: folhas; raízes (nódulos na soja) e parte aérea. Foram determinados nas folhas utilizadas para diagnose em soja e girassol: os teores de macro e micronutrientes, as atividades da redutase do nitrato e da urease e as concentrações dos ácidos orgânicos: oxálico, malônico, succínico, málico, tartárico, fumárico, oxaloacético, cítrico e lático. Os mesmos ácidos orgânicos foram determinados em raízes secundárias de girassol e nódulos de soja. Foram realizadas avaliações ultraestruturais por meio de microscopia eletrônica de transmissão (MET) em raízes de girassol, e estruturais e de tonalidade em nódulos de soja, por meio de microscopia de luz. No solo, foram determinadas: atividade urease, desidrogenase, Ni total e fitodisponível pelos métodos: Mehlich-1, Mehlich-3 e DTPA. No período de maturidade fisiológica de cada cultura foi realizada a colheita das plantas dos vasos restantes para determinação de produção de grãos, teores de Ni na planta inteira e Ni e N nos grãos. Ao final dos dois experimentos foi realizada nova coleta de solo para extração sequencial de Ni. O índice SPAD em soja aos 60 d.a.e., a produção de massa seca da parte aérea da soja e da raiz de girassol foram influenciados pela saturação por bases, doses de níquel e pela a interação destes. Foram influenciados pelas saturações por base e doses de níquel (fatores isolados): para soja: AP aos 60 d.a.e., NN aos 30 e 60 d.a.e., SPAD aos 30 d.a.e.; para girassol: AP e NN aos 30 e 60 d.a.e., DC e SPAD aos 30 d.a.e. As demais variáveis avaliadas aos 30 e 60 d.a.e. foram influenciadas apenas pela saturação por bases, ou doses de Ni separadamente. As plantas de soja e girassol apresentaram maiores teores de Ni nos diferentes tecidos avaliados (exceto grãos) quando cultivadas sob V50%. A produção de grãos de soja e girassol não foi influenciada pelos tratamentos, porém o teor de N dos grãos de soja influenciado pelas doses de Ni na V70%. A atividade da enzima urease nas folhas de soja e girassol foi responsiva positivamente ao aumento das doses de Ni. Quatro dos ácidos orgânicos avaliados e o teor de N nas folhas e nos grãos foram maiores nas plantas cultivadas sob V70% com a dose de 0,5 mg dm-3 de Ni. As doses de Ni bem com as saturações por bases influenciaram diretamente o balanço de nutrientes das plantas. Os extratores Mehlich-1, Mehlich-3 e DTPA apresentaram elevado coefienciente de correlação entre a fração de Ni disponível no solo e a concentração do elemento nas plantas de soja e girassol, sendo o extrator DTPA o que apresentou maior coeficiente de correlação. O Ni apresentou distribuição variável entre as diferentes frações do solo em função dos tratamentos. Os solos dos tratamentos com saturação por bases de 70% apresentaram maior concentração de Ni ligado a carbonato, comparado aos tratamentos sob saturação por bases de 50%. A distribuição do Ni entre as frações do solo seguiu a seguinte orgem: ligado a carbonato < trocável < ligado a óxidos < matéria orgânica < residual. A saturação por bases exerceu efeito diferenciado para a atividade da urease no solo em função da cultura avaliada. Por sua vez, o Ni exerceu efeito diferenciado sobre a atividade de desidrogenase em função da cultura estudada / As an essential element for plants and a heavy metal at the same time, nickel requires attention on the aspects of the physiology of plants and environmental. Furthermore, there is a narrow range between nutritional requirements and toxic levels to plants. In this context, the aim was to evaluate the effect of Ni in soil-plant system through experiment under controlled conditions, using soil pots distributed entirely at random, using a factorial 2 x 5, with seven repetitions each treatment. The first factor was formed of two saturations base (50 and 70%) and the second of five Ni rates (0, 0.1, 0.5, 1.0 and 10.0 mg dm-3 of soil). The pots were filled with 8 dm3 of soil and sown with soybean [Glycine max (L.) Merrill] succeeded by sunflower (Helianthus annuus L.). The agronomic traits: plant height (PH), stem diameter (SD), number of nodes (NN), phenological stage (PS), SPAD index, head diameter (HD) (Sunflower) were evaluated at 30 and 60 days after emergence (dae) of each crop. Whole soybean plants, sampled in four pots of each treatment were collected at the R1 stage for analysis of soil samples from the rhizosphere. Then, the collected plants were divided into: sheets; roots (nodules on soybean) and shoot. Were determined in the leaves used for diagnosis in soybean and sunflower: macro and micronutrients, activities of nitrate reductase, and urease and concentrations of organic acids: oxalic, malonic, succinic, malic, tartaric, fumaric, oxaloacetic, citric and lactic. The same organic acids were determined in secondary roots of sunflower and nodules of soybean. They were conducted ultrastructural evaluations by means of transmission electron microscopy (TEM) in sunflower roots, and color in soybean nodules, using light microscopy. In the soil were determined: urease activity dehydrogenase, the total Ni and available by methods: Mehlich-1, Mehlich-3 and DTPA. At physiological maturity period of each culture was performed to harvest the plants of other pots in order to determine grain dry matter yield, Ni content in the whole plant and Ni and N content in grains. At the end of the two experiments it were carried out new soil sampling for Ni sequential extraction. The SPAD index taken on soybean leaves at 60 d.a.e., the dry matter production of the aerial part of soybean and sunflower root were influenced by the base saturation, nickel doses and the interaction among them. They were influenced by the saturation for base and nickel doses (single factor): soybean: HP d.a.e. to 60, NN at 30 and 60 d.a.e., SPAD to 30 d.a.e.; Sunflower: PH and NN at 30 and 60 d.a.e., HD and SPAD to 30 d.a.e. The other variables assessed at 30 and 60 d.a.e. They were influenced only by base saturation, or Ni doses separately. Sunflower soybean plants had higher Ni contents at the different tissues (except grain) when grown under V50%. The production of soybean and sunflower grains was not affected by the treatments, but the N content of the soybeans was changed by Ni rates V70%. The urease enzyme activity in soybean and sunflower leaves was responsive to increased doses of Ni. Four of the organic acids and the N content in leaves and grains were higher in plants grown under V70% at the dose of 0.5 mg dm-3 Ni. Ni rates as well as the cation base saturation directly influenced the balance of plant nutrients. The Mehlich-1, Mehlich-3 and DTPA presented higher coefiencient correlation between available Ni soil fraction with nutrient concentrations in both soybean and sunflower, with the DTPA solution showing the highest correlation coefficient in this analysis. Ni showed variable distribution among different soil fractions in the treatments. The treatments presented soil under cation bases saturation of 70% showed higher Ni concentration linked to carbonate, as compared to treatments under cation base saturation of 50%. The distribution of Ni between the soil fractions followed the following order: bound to carbonate <exchangeable < bound to oxides <organic matter <residual. The cation base saturation showed different effects on the urease activity according to the plant evaluated. Ni exerted on the differential effect dehydrogenase activity according to the plant specie

Com o objetivo de estabelecer as possíveis correlações entre os ácidos orgânicos na precipitação e as alterações no uso do solo, amostras de água da chuva e aerossol foram coletadas nos Parques Estaduais de Intervales (PEI) e Morro do Diabo (PEMD), no Estado de São Paulo, cuja vegetação é formada por florestas pertencentes ao domínio Mata Atlântica. Entre julho de 2002 a julho de 2004 foram coletadas 232 amostras de água de chuva em PEI e 96 amostras em PEMD, determinando-se os teores de formato, acetato, metanosulfonato, piruvato, oxalato, maleato, valerato, citrato, butirato, tartarato, cloreto, nitrato, sulfato, sódio, amônio, potássio, cálcio e magnésio por cromatografia iônica (CI). Também foram determinadas as concentrações de carbono orgânico dissolvido (COD), carbono inorgânico dissolvido (CID) e o pH das amostras. Entre julho de 2002 a maio de 2003, amostras de aerossol inalável (PM10), subdividido nas frações fina (dp < 2,5 µm) e grossa (2,5 < dp < 10 µm), foram coletadas separadamente, totalizando 60 amostras para cada fração no PEMD e 90 amostras para cada fração no PEI, determinando-se as concentrações em massa por gravimetria e a composição química do material solúvel em água por CI. Eventos de chuva ácida ocorreram nos dois locais ao longo do período de amostragem. Os ácidos orgânicos contribuíram com até 16% da somatória iônica (µeq.L-1) no PEI e até 12% no PEMD, com predominância do ácido acético no PEI e com predominância de ácido fórmico no PEMD. A emissão direta da vegetação foi a principal fonte de ácidos orgânicos no PEI, uma vez que a razão entre o ácido fórmico e ácido acético < 1,0. No PEMD, a razão entre ácido fórmico e acético > 1,0 indicou como as principais fontes as reações secundárias de oxidação fotoquímicas no local e a emissão direta biogênica da vegetação. A contribuição dos ácidos orgânicos para a acidez da água da chuva foi de até 40% no PEI e até 30% no PEMD. Adicionalmente, as correlações significantes entre H+ e COD (p < 0,05) sugerem contribuições de compostos orgânicos para acidez da água da chuva. A influência de fontes regionais de queima de biomassa e das condições meteorológicas na composição química da atmosfera no PEMD é evidenciada pela concentração média de PM10 (22 ug.m-3); pela concentração de \"black carbon\" (40 ?g.m-3), superiores às verificadas no PEI, pela maior contribuição da fração fina do aerossol para a concentração de COD da fração solúvel em água. Por outro lado, no PEI, a fração grossa relacionada à compostos biogênicos, aerossol marinho e partículas de solo foi a maior contribuinte para o COD. Dos ácidos orgânicos detectados no aerossol solúvel em água, o ácido oxálico foi o mais abundante. / In order to establish the possible correlations among organic acids in the precipitation and land use changes, rain water and aerosol samples were collected in the State Parks of Intervales (PEI) and Morro do Diabo (PEMD), in the State of São Paulo. The vegetation there is formed by Mata Atlântica forests (Atlantic forest). Between July, 2002 and July, 2004, 232 samples of rainwater were collected in PEI and 96 samples in PEMD, for the analysis of formate, acetate, methanesulfonate, pyruvate, oxalate, malate, valerate, citrate, butyrate, tartarate, chloride, nitrate, sulfate, sodium, ammonium, potassium, calcium, magnesium for ionic chromatography (CI). The concentrations of dissolved organic carbon (COD), dissolved inorganic carbon (CID) and pH of the samples were also determined. Between July, 2002 and May, 2003, samples of inhalable aerosol (PM10), subdivided in fine fraction (dp <2.5 µm) and coarse fraction (2.5 <dp <10 µm), were collected separately, in a total of 60 samples for each fraction in PEMD and 90 samples for each fraction in PEI, submitted to the analyses of concentration in mass through gravimeter and the chemical composition of the water soluble aerosol for CI. Events of acid rain were verified along the sampling period in both places and the organic acids contributed up to 16% of the ionic sum (µeq.L-1) in PEI and up to 12% in PEMD, with predominance of acetic acid in PEI and with predominance of formic acid in PEMD. The direct emission from vegetation was the main source of organic acids in PEI, once the ratio between the formic acid and acetic acid was <1.0 and that the effects of biomass burns are not verified. In PEMD, the ratio among formic and acetic acid >1.0 indicated the secondary reactions of photo-chemical oxidation ?in situ? and biogenic direct emission from the vegetation as the main source. The contribution of the organic acids for the acidity of the rainwater was up to 40% in PEI and up to 30% in PEMD. In addition to the strong correlations between H+ and COD (p <0.05) they suggest contributions of organic compositions for the acidity of the rainwater. The influence of regional sources of biomass burning and of the meteorological conditions in the chemical composition of the atmosphere in PEMD is also observed by the medium concentration of PM10 (22 µg.m-3) and concentration of \"black carbon\" (40 ?g.m-3), both higher than the ones observed in PEI, and for the highest contribution of fine fraction of aerosol for the concentration of COD analyzed from the water soluble fraction. In PEI, the coarse fraction related to biogenic compounds, marine aerosol and soil particles were the greatest responsible agents for COD. Oxalic acid was the most abundant organic acid detected.

A salmonelose é uma das mais importantes causas de toxinfecção alimentar em humanos, provocando sérios problemas à saúde pública, além de ser uma das doenças de maior impacto na produção avícola. Em vista disso, este estudo objetivou avaliar o uso de ácido fórmico no controle de Salmonella Enteritidis (SE). Assim, foram utilizados 360 pintinhos de um dia de idade: 50% das aves foram inoculadas via inglúvio com 3x107 UFC/0,1mL de SE e 50% não (aves contactantes). As aves foram divididas em cinco (05) grupos com seis (06) repetições cada: T1) controle sem o aditivo; T2) ácido fórmico 0,3% na ração; T3) ácido fórmico 0,2% na ração; T4) ácido fórmico diluído em água de beber até que a mesma atingisse um pH 4; T5) ácido fórmico 0,2% protegido na ração. As aves receberam ácido do 1º ao 7º dia e do 35º ao 42º dia. A cada sete dias, amostras de fezes foram colhidas da cama em todos os grupos para se avaliar o isolamento de SE. Aos 42 dias foram colhidas à necropsia amostras de inglúvio, fígado/baço e ceco para o isolamento de SE e sangue para a avaliação sorológica. Nas aves do T1, do T2 e do T5 SE foi isolada do inglúvio, fígado/baço e ceco das aves inoculadas e das contactantes. Houve isolamento de SE no T3 apenas em inglúvio e ceco das aves inoculadas e, no T4, apenas do ceco das contactantes. Em todos os grupos SE foi reisolada do material da cama, embora não em todas as semanas. A avaliação sorológica realizada com teste comercial de ELISA revelou aves sororeagentes no T1 (inoculadas e contactantes), no T2 (inoculadas) e no T3 (contactantes). Nenhuma ave foi sororeagente no T4 e T5. A transmissão horizontal foi evidenciada em todos os grupos estudados. Os dados foram comprovados por ANOVA e teste LCD. Nos grupos T3 e T4 o ácido fórmico foi eficaz em reduzir a presença e a transmissão horizontal de SE. / Salmonellosis is one of the most common and widely distributed foodborne diseases, with tens of millions of human cases occurring worldwide every year. Furthermore, this disease is responsible to significant economic losses to the poultry industry. Endemic infection of broiler chickens with Salmonella. Enteritidis (SE) and consequential human illness has been well documented. For these reasons the aim of this study was to evaluate the use of formic acid to control SE in broilers experimentally infected. Thus 360 day-old chicks were used in the experiment: 50% (180) of birds were inoculated in crop with 3x107CFU/0.1 mL of SE and 50% (180) were used as controls and kept in contact with the infected chicks. For the formic acid treatment the birds were divided into five (05) groups with six (06) replicates as follows: T1) control without additive; T2) 0.3% formic acid in the diet ; T3) 0.2% formic acid in the diet ; T4) formic acid diluted in drinking until it reached pH 4 water; T5) 0.2% formic acid microencapsulated in the food. The birds received the formic acid from the 1st to 7th day and from the 35th to 42nd days. Every seven days faeces samples were collected from the chick\'s litter and SE isolation was attempted. In the 42 day blood samples were collected for serological evaluation by means of Elisa. Additionally samples from the crop, liver/spleen and cecum were collected at necropsy and submitted to culture for SE detection. SE was isolated from crop, liver/spleen and cecum of the inoculated and contacts birds belonging to the T1, T2 and T5. Regarding the T3, SE was isolated in the crop and cecum of the inoculated birds and in the T4 only in the contacts chicks cecum. In all groups SE was isolated from the litter, although not in every week. Serological evaluation revealed serum positive birds in T1 (inoculated and contacts), T2 (inoculated) and T3 (contacts). Antibodies anti-SE were not detected in the serum from the birds of T4 and T5. Horizontal transmission was observed in all groups. The data were confirmed by ANOVA and LCD test. In groups T3 and T4 formic acid was effective in reducing the presence and horizontal transmission of SE.

A nêspera (Eriobotrya japonica Lindl.) é uma fruta do tipo pomo, que tem sido cultivada principalmente no estado de São Paulo. Cinco cultivares de nêspera foram selecionadas por se destacarem no sabor e na capacidade produtiva. Foi realizado o estudo das mudanças fisiológicas e físico-químicas dos frutos durante o desenvolvimento e o período pós-colheita. Amostras das cultivares (Centenária, Mizuho, Mizumo, Néctar de Cristal e Mizauto) foram colhidas semanalmente conforme o avanço do estádio de desenvolvimento (na planta), desde a fase verde dos frutos até se tornarem maduros. Os frutos maduros para estudo do período pós-colheita foram colhidos seguindo critérios adotados por produtores, através da cor dos frutos. As determinações físico-químicas realizadas nas nêsperas maduras foram umidade, fibra, ácido ascórbico, açúcares solúveis (glicose, frutose, sacarose e sorbitol), ácidos orgânicos e carotenóides. Durante o período de desenvolvimento das nêsperas, as curvas de respiração decresceram progressivamente, e houve um considerável aumento na produção de etileno, precedendo o aumento na concentração de açúcares e as mudanças de coloração dos frutos (verde a amarela ou alaranjada). A composição de açúcares e ácidos dos frutos maduros foi semelhante a encontrada nos frutos precocemente maduros durante o desenvolvimento na planta. Portanto, a observação visual da cor não foi apropriada para a definição do ponto ideal de colheita. Durante o período póscolheita, as curvas de respiração e de produção de etileno das cultivares tenderam a queda, mostrando comportamento típico de frutos não-climatéricos. O estudo fisiológico e físico-químico das cinco cultivares de nêspera permitiu conhecer melhor o padrão de amadurecimento de cada cultivar, e assim obter informações que auxiliem na definição do ponto ideal de colheita, e que permitam obter frutos de boa qualidade e apropriados para distribuição comercial. / Loquat (Eriobotrya japonica Lindl.) fruit is a pome fruit cultivated mainly in the state of São Paulo. Five loquat fruit cultivars were selected to analysis mainly by their high productivity and taste. Physiological and physico-chemical changes were analised during development and ripening fruit. Samples were harvest weekly according to progress in development stage (in the tree), since imature stage (green) until mature stage. Mature stage to harvest was defined by fruit color, like methods\' producer. The physico-chemical parameters analysed were moist value, fiber, ascorbic acid, soluble sugars (glucose, fructose, sucrose and sorbitol), organic acids and carotenoids. During development of loquat fruits, the respiration levels decreased, and show a increase in ethylene levels, which was associed with increase of sugar concentration in the pulp and the changes of fruit color (green to yellow or orange color). According to sugars and acids levels during development (in the tree) and in ripe stage, the composition in ripe fruits was similar to that early mature fruits analyzed during development in the tree. The visual determination of color was not appropriate to define the harvest point of fruits. During postharvest period of the cultivars, the levels in respiration and ethylene production decreased, which is a typical pattern of non-climacteric fruits. The study of physiological and physico-chemical characteristics of loquat cutivars contribute to know the ripening pattern of each cultivar and to auxiliary in the definition of best mature point to harvest and to comercial distribution.

The effects of crop load and berry exposure on the composition of Marlborough Sauvignon blanc grapes and wine from the Brancott vineyard, Blenheim, were explored. Commercially grown, 2-cane and 4-cane Sauvignon blanc vines were used with a row orientation of north-south. Two exposure treatments were imposed in the following manner: complete leaf removal was undertaken in the fruit zone and 50% shade cloth was erected to give a uniform shading treatment to half the trial vines. Weekly thirty-berry and whole bunch samples were taken from each of the 32 plots with the exception of the veraison period when two samples per week were taken. Vine vigour was assessed using pruning and leaf area per vine data. Harvest occurred on different dates for 2-cane and 4-cane pruned vines so that fruit attained from both treatments had similar °Brix. Fruit was processed at the Lincoln University winery. Must analysis and wine analysis were undertaken. As expected, 4-cane vines had almost double the yield of 2-cane vines. Higher crop load significantly reduced leaf area per shoot and shoot thickness. Lower leaf area to fruit ratio for 4-cane berries resulted in delayed onset of veraison and slowed the rate of sugar accumulation. Crop load, which limited leaf area to fruit ratio, appeared to be the dominant factor in determining timing of grape physiological ripeness as expressed by °Brix over other factors such as fruit exposure. Malic acid, tartaric acid, IPMP (iso-propylmethoxypyrazine) and IBMP (iso-butyl-methoxypyrazine) were lower at equivalent °Brix in 4-cane compared with 2-cane berries. Significantly higher concentrations of quercetin were found in exposed compared to shaded berries. Must analysis showed a significant influence of crop load on berry titratable acidity and pH, reflecting berry ripening results. Exposure significantly increased the concentrations of nitrogenous compounds in 4-cane must yet showed no influence on 2-cane must. After wine processing lower malic acid concentrations in wines made from 100% exposed fruit became evident in lower wine titratable acidity but showed no influence on wine pH. Bentonite addition to wines had a small but statistically significant influence on wine by reducing pH, titratable acidity and alcohol. Bound sulphur concentrations were significantly higher in 4-cane versus 2-cane wines. At harvest, methoxypyrazine levels in grapes and wines were very low; IBMP concentrations where significantly lower than those normally found in Sauvignon blanc wines from Marlborough. This was attributed to the absence of basal leaves from the shoots of ripening berries. The results suggest that leaf area to fruit ratio is a powerful determinant of grape and wine quality.

A rapid and accurate quantitative method was developed and validated for the analysis of four urinary organic acids with nitrogen containing functional groups, formiminoglutamic acid (FIGLU), pyroglutamic acid (PYRGLU), 5-hydroxyindoleacetic acid (5-HIAA), and 2-methylhippuric acid (2-METHIP) by liquid chromatography tandem mass spectrometry (LC/MS/MS). The chromatography was developed using a weak anion-exchange amino column that provided mixed-mode retention of the analytes. The elution gradient relied on changes in mobile phase pH over a concave gradient, without the use of counter-ions or concentrated salt buffers. A simple sample preparation was used, only requiring the dilution of urine prior to instrumental analysis. The method was validated based on linearity (r2 ¡Ý 0.995), accuracy (85¨C115%), precision (C.V. < 12%), sample preparation stability (¡Ü 5%, 72h), and established patient ranges. The method was found to be both efficient and accurate for the analysis of urinary zwitterionic organic acids.

Low molecular weight organic acids (LMWOAs) are important nutrients for microbes. However, most LMWOAs do not exist freely in the environment but are bound to macromolecular organic matter, e.g. kerogen, lignite and coal. During burial and geological maturation of sedimentary macromolecular organic matter biological and abiological processes promote the liberation of LMWOAs into the surrounding sediment. Through this process, microbes in sedimentary subsurface environments are supplied with essential nutrients. To estimate the feedstock potential of buried macromolecular organic matter to many environments it is important to determine the amount of LMWOAs that are bound to such a matrix. However, high-pressure and high temperature are a key feature of deep subsurface environments, and these physical parameters have a profound influence on chemical reaction kinetics. Therefore it is essential for the estimation of the feedstock potential to generate high-pressure and high temperature for the liberation of LMWOAs to recreate true in-situ conditions. This work presents a newly developed, inexpensive incubation system for biological and geological samples. It allows the application of high-pressure and high temperature as well as a subsampling of the liquid phase without loss of pressure, thereby not disturbing the on-going processes. When simulating the liberation of LMWOAs from sedimentary organic matter, the newly developed incubation system produces more realistic results than other extraction systems like Soxhlet. The extraction products remain in the extraction medium throughout the extraction, influencing the chemical conditions of the extraction medium. Sub-bituminous coal samples from New Zealand as well as lignite samples from Germany were extracted at elevated temperature (90˚C) and pressure (5 MPa). The main LMWOAs released from these low rank coals were formate, acetate and oxalate. Extraction efficiency was increased by two to four times for formate, acetate and oxalate in comparison to existing extraction methods without pressurisation and with demineralised water. This shows the importance of pressure for the simulation of true in-situ conditions and suggests that the amount of bioavailable LMWOAs is higher than previously thought. With the increase in carbon capture and storage (CCS) and the enhanced recovery of oil and gas (EOR/EGR), more and more CO2 becomes injected into the underground. However, the effects of elevated concentrations of carbon dioxide on sedimentary organic matter are rarely investigated. As the incuabtion system allows the manipulation of the composition and partial pressure of dissolved gasses, the effect of highly gas-enriched (CO2, CO2/SO2, CO2/NO2; to simulate flue gas conditions) waters on the extraction yield of LMWOAs from macromolecular organic matter was evaluated. For sub-bituminous coal the concentrations of all LMWAOs decreased upon the addition of gas, irrespective of its composition, whereas for lignite formate always and acetate mostly increased, while oxalate decreased. This suggests an positive effect on the nutrient supply for the subsurface microbiota of lignite layers, as formate and acetate are the most common LMWOAs used for microbial metabolism. In terrestrial mud volcanoes (TMVs), sedimentary material is rapidly ascending from great depth to the surface. Therefore LMWOAs that were produced from buried macromolecular organic matter at depth are also brought up to the surface, and fuel heterotrophic microbial ecosystems at the surface. TMVs represent geochemically and microbiologically diverse habitats, which are supplied with organic substrates and electron acceptors from deep-seated hydrocarbon-generating systems and intersected shallow aquifers, respectively. The main electron donor in TMVs in Azerbaijan is sulphate, and microbial sulphate reduction leads to the production of a wide range of reduced sulphur species that are key players in several biological processes. In our study we estimated the effect of LMWOAs on the sulphur metabolising activity of microorganims in TMVs from Azerbaijan. The addition of a mixture of volatile fatty acids containing acetate and other LMWOAs showed significant positive response to the sulphate reduction rate (SRR) of samples of several mud volcanoes. Further investigations on the temperature dependency of the SRR and the characterisation of thermophilic sulphate-reducing bacteria (SRB) showed a connection between the deep hot subsurface and the surface. / Niedermolekulare organische Säuren (nachfolgend als LMWOAs - low molecular weight organic acids - bezeichnet) stellen wichtige mikrobielle Substrate dar. Jedoch liegen die meisten LMWOAs nicht in freier, bioverfügbarer Form vor, sondern sind vielmehr an hochmolekulare organische Substanzen gebunden, z.B. Kerogen, Lignit und Kohle. Während der geologischen Verbringung in tiefe Erdschichten und der geologischen Reifung von sedimentären hochmolekularen organischen Substanzen, führen biologische und abiologische Prozesse zu einer Freisetzung von LMWOAs in die umgebenden Sedimente. Durch diesen Prozess werden Mikroorganismen in unterirdischen sedimentären Ökosystemen mit essentiellen Nährstoffen versorgt. Um das Nährstoffpotential tief liegender hochmolekularer organischer Substanzen für diverse Ökosystemen abschätzen zu können, ist es notwendig, die Menge an LMWOAs, die an solch eine hochmolekulare Matrix gebunden ist, zu bestimmen. Dabei stellen hoher Druck sowie hohe Temperatur entscheidende Faktoren in tiefen unterirdischen Ökosystemen dar, welche einen signifikanten Einfluss auf chemische Reaktionen haben. Daher ist es für die Abschätzung des Nährstoffpotentials entscheidend, hohen Druck und hohe Temperatur bei der Freisetzung von LMWOAs zu erzeugen, um wahre in situ Bedingungen zu schaffen. In der vorliegenden Arbeit wird ein neu entwickeltes, preiswertes Inkubationssystem für biologische und geologische Proben präsentiert. Es erlaubt die Verwendung von hohem Druck als auch hoher Temperatur sowie eine Unterprobennahme der flüssigen Phase ohne Druckverlust, um den fortlaufende Prozess nicht zu unterbrechen. Bei der Simulierung der Freisetzung von LMWOAs aus sedimentären organischen Substanzen erhält man mit dem neu entwickelten Inkubationssystem realistischere Resultate als mit anderen Extraktionssystemen, wie z.B. eine Soxhlet-Apparatur. Die Extraktionsprodukte verbleiben während der Extraktion im Extraktionsmedium, wodurch die chemischen Bedingungen verändert werden. Kohleproben aus Neuseeland sowie aus Deutschland wurden mittels erhöhter Temperatur (90°C) und Druck (5 MPa) extrahiert. Die wichtigsten LMWOAs, die aus diesen Kohlen freigesetzt wurden, waren Formiat, Acetat und Oxalat. Die Extraktionseffizienz für diese LMWOAs konnte im Vergleich zu existierenden Extraktionsmethoden ohne Druck um den Faktor 2 bis 4 gesteigert werden. Dies zeigt die Bedeutung von Druck bei der Simulation von in situ Bedingungen und legt nahe, dass die Menge an bioverfügbaren LMWOAs größer ist als bisher angenommen. Durch die Zunahme der CO2-Speicherung im Untergrund (carbon capture and storage, CCS) sowie der erweiterten Förderung von Öl und Gas (enhanced recovery of oil and gas, EOR/EGR) wird immer mehr CO2 in den Untergrund gepresst. Jedoch sind die Auswirkungen von erhöhten CO2-Konzentrationen auf sedimentäre organische Materie noch unerforscht. Da mit dem Inkubationssystem die Veränderung der Zusammensetzung und des Partialdruckes von gelösten Gasen möglich ist, wurde der Effekt von hoch mit Gasen (CO2, CO2/SO2, CO2/NO2; um Kraftwerksabgase zu simulieren) angereicherten Wässern auf die Extraktionsausbeute von LMWOAs untersucht. Bei der subbituminösen Kohle zeigte sich eine Abnahme aller LMWOAs-Konzentrationen durch die Lösung von Gas im Extraktionsmedium, wobei die Art des Gases keine Rolle spielte. Bei Lignit konnte hingegen festgestellt werden, dass die Extraktionsausbeute an Formiat immer und an Acetat meistens erhöht wurde, während sie sich bei Oxalat verringerte. Dies deutet auf einen positiven Effekt für die Nährstoffversorgung von Mikroorganismen um Lignit-Lagerstätten an, da Formiat und Acetat die am häufigsten verwendeten LMWOAs im mikrobiellen Stoffwechsel darstellen. In terrestrischen Schlammvulkanen (terrestrial mud volcanoes, TMVs) steigt sedimentäres Material aus großen Tiefen an die Erdoberfläche. Somit werden auch LMWOAs, welche aus hochmolekularen organischen Substanzen freigesetzt werden, an die Oberfläche verbracht, und ermöglichen dort heterotrophe Ökosysteme. TMVs stellen dabei geochemisch und mikrobiell unterschiedliche Habitate dar, welche mit organischen Substraten und Elektronenakzeptoren aus tief liegenden, Kohlenwasserstoffe erzeugenden Systemen versorgt werden. In TMVs in Aserbaidschan stellt Sulfat den Hauptelektronenakzeptor dar, wobei mikrobielle Sulfatreduktion zu einer Vielzahl an reduzierten Schwefelspezies führt. In der vorliegenden Arbeit wurde der Effekt von LMWOAs auf die Aktivität von Mikroorganismen bei der Umsetzung von Schwefel in TMVs in Aserbaidschan untersucht. Die Zugabe einer Mischung verschiedener kurzkettiger Fettsäuren zu Schlammproben verschiedener TMVs erzeugte eine signifikant positive Reaktion in Bezug auf die Sulfat-Reduktionsraten. Weiterführende Untersuchungen zur Temperaturabhängigkeit der Sulfat-Reduktionsraten und die Charakterisierung thermophiler, Sulfat-Reduzierender Bakterien zeigte eine Verbindung zwischen der tiefen, heißen Biosphäre und der Erdoberfläche auf.