Submitted by Jaqueline Silva (jtas29@gmail.com) on 2014-10-06T19:08:19Z No. of bitstreams: 2 Tese - Jussara de Souza Carneiro - 2013.pdf: 1644698 bytes, checksum: 3faeafe59bfc34f56dc772cbdb1cf640 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2014-10-06T19:15:39Z (GMT) No. of bitstreams: 2 Tese - Jussara de Souza Carneiro - 2013.pdf: 1644698 bytes, checksum: 3faeafe59bfc34f56dc772cbdb1cf640 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-10-06T19:15:39Z (GMT). No. of bitstreams: 2 Tese - Jussara de Souza Carneiro - 2013.pdf: 1644698 bytes, checksum: 3faeafe59bfc34f56dc772cbdb1cf640 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-08-01 / Aimed at feeding laying hen, corn has been substituted by other cheaper food. However, since these foods have little capacity to stain egg yolk, pigments are usually added to the animal’s feed. Synthetic carotenoids, such as canthaxanthin, and the natural pigments present in bran annatto are among the most used food coloring. However, little is known about the impact of those pigments to the animal’s health, and it is important to point out that the choice of the dye and its dose are based solely on the eggs color. In the present study we aimed at evaluating if the pigments have some beneficial or toxic effects on the laying hen health, and test a new simple, fast and non subjective computerized method to analyze the egg color. Thus, laying hens of the ISA Brown lineage were fed with three different sorghum meal: without dye; with bran annatto (3%); or with cathaxanthin (8 ppm) + apocaroten (25 ppm). Animal performance, eggs quality, the liver and kidney functions, as well as the oxidative stress inferred from the levels of malondialdehyde in red blood cells were determined. None of the treatments altered the laying hen performance, the liver and kidney integrity and functions, and the oxidative stress. However, all the dyes tested increased yolks egg weight and color. Finally, the results clearly suggest that yolks color intensity was more accurately determined by the computerized method when compared to the traditional colorimetric fan assay. / O milho vem sendo substituído por outros alimentos na alimentação de galinhas poedeiras. Contudo, como tais alimentos apresentam baixa capacidade de corar as gemas dos ovos, pigmentantes são comumente adicionados à ração dos animais. Carotenóides sintéticos como cantaxantina, bem como pigmentos naturais encontrados no farelo de urucum estão entre os mais utilizados. Contudo, sabe-se muito pouco sobre o impacto da utilização de tais pigmentantes na saúde das galinhas, além da determinação da dose ser baseada somente na coloração das gemas dos ovos. No presente estudo avaliou-se se os pigmentantes apresentam algum efeito sobre a saúde das galinhas, bem como testou-se um novo método computadorizado, simples, rápido e não subjetivo, para a avaliação da coloração das gemas. Assim, galinhas poedeiras vermelhas foram alimentadas com três diferentes rações a base de sorgo: sem pigmentante; com 3% de farelo de urucum ou com cantaxantina (8 ppm) + apocaroteno (25 ppm). Foram avaliados o desempenho das galinhas, a qualidade dos ovos, as funções renais e hepáticas, e o estresse oxidativo (inferido a partir dos níveis de malondealdeído de eritrócitos). Nenhum dos tratamentos alterou o desempenho das galinhas, a qualidade dos ovos e as funções hepáticas e renais. Contudo, todos os corantes aumentaram a coloração e o peso das gemas dos ovos. Por fim, os resultados sugerem claramente que a intensidade da coloração das gemas foi determinada com maior precisão pelo método computadorizado quando comparado ao ensaio colorimétrico tradicional.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-08-30T14:26:59Z No. of bitstreams: 1 2010 - Fernanda Peixoto Giori.pdf: 2362278 bytes, checksum: a15d423533272c981b15d7f22b714be1 (MD5) / Made available in DSpace on 2016-08-30T14:26:59Z (GMT). No. of bitstreams: 1 2010 - Fernanda Peixoto Giori.pdf: 2362278 bytes, checksum: a15d423533272c981b15d7f22b714be1 (MD5) Previous issue date: 2010-02-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior-CAPES / Brazil was adverted by the World Health Organization (WHO) as a sub-clinical area of serious vitamin A deficiency. Thus, the Brazilian Agricultural Research Corporation (EMBRAPA) is selecting and improving varieties of sweet potatoes with higher levels of Beta-carotene, pro-vitamin A. The carotenoids provitamin A amount of in foods does not necessarily correspond the amount that is absorbed and metabolized by the body. For a better determination of these values and knowledge of the mechanisms of its transport and absorption, it is necessary to understand the factors that lead to the food matrix release, until the absorption and the influence on the promotion and maintenance in human health. In order to perform preliminary studies of its absorption, this study aims to determine the efficiency of micellization of Beta -carotene in orange sweet potato (Ipomoea batatas, Lam), by applying a in vitro digestion as a tool for determining the bioaccessibility, which is the first step for bioavailability determination. This approach aims to simulate the oral, gastric and intestinal stages of human. The digestion was performed with 10 g of fresh samples, homozeneided with 5% (w/w) of canola oil. The extraction was performed with acetone and petroleum ether and the micellar fraction with petroleum ether, NaCl 10% (w / v) and NaSO4 2% (w/v) and involves the use of enzymes as -amylase , pepsin, bile, pancreatin, lipase and mucin, and inorganic compounds such as KCl, KSCN, NaH2PO4, Na3PO4, NaOH, NaCl, CaCl2, HCl, NaHCO3. The physiological variations are reproduced by the heating bath shaker with orbital gyrus (37?C) and centrifugation (5000g, 45 min). Quantification and determination of the profile of carotenoids were performed by high performance liquid chromatography (HPLC) with YCM ? C30 Carotenoid S-3 4.6 x 250mm column. Quantification of total carotenoids was performed by UV-VIs. The whole procedure was performed under controlled temperature (25 ? C) and light. The Beta -carotene was present mainly with levels of 86%, 73% and 82% for access 1, 2 and 3 and after digestion, the profile of Beta -carotene has set levels of 96%, 89% and 100%, respectively. The efficiency of micellization was 23.8%, 28% and 25% for 1.2 and 3 hits, indicating Beta -carotene transfer of the food matrix to micelles, corresponding to bioaccessibility of the compound. This methodology proved to be faster and cheaper, since the in vivo studies are costly, complex and require more time. / O Brasil foi classificado pela Organiza??o Mundial da Sa?de (OMS) como ?rea de car?ncia sub-cl?nica grave de vitamina A. Assim, a Empresa Brasileira de Pesquisa Agropecu?ria (EMBRAPA) v?m selecionando e melhorando variedades de batata-doce com teores maiores de Beta-caroteno, composto pr?-vitamina A. A quantidade de caroten?ides pr?-vitamina A presentes nos alimentos n?o corresponde necessariamente ?quela quantidade absorvida e metabolizada pelo organismo. Para uma melhor determina??o destes valores e conhecimento dos mecanismos de transporte e absor??o deste composto, faz-se necess?rio, o entendimento dos fatores que levam ? sua libera??o da matriz do alimento, at? a extens?o de sua absor??o, bem como a influ?ncia na promo??o e manuten??o da sa?de humana. A fim de realizar estudos preliminares de sua absor??o, este trabalho visa determinar a efici?ncia de miceliza??o de Beta-caroteno de batata-doce de polpa alaranjada (Ipomoea batatas, Lam.), atrav?s da aplica??o de digest?o in vitro, como ferramenta de determina??o da bioacessibilidade, etapa preliminar para a determina??o da biodisponibilidade. Esta metodologia visa simular as etapas de digest?o oral, g?strica e intestinal humana. Foram pesados 10g de amostra in natura e adicionados 5% (p/p) de ?leo de canola. A extra??o do alimento foi realizada com acetona e ?ter de petr?leo e a da fra??o micelar, com ?ter de petr?leo, NaCl 10%(p/v) e NaSO4 2%(p/v) . Enzimas como: a-amilase, pepsina, bile, pancreatina, lipase e mucina, bem como compostos inorg?nicos, tais como KCl, KSCN, NaH2PO4, Na3PO4, NaOH, NaCl, CaCl2, HCl, NaHCO3. As varia??es fisiol?gicas foram reproduzidas pelo banho de aquecimento com giro orbital (37?C) e centrifuga??o (5000g;45 min). A quantifica??o e determina??o do perfil de caroten?ides foi realizada por cromatografia l?quida de alta efici?ncia (CLAE), com coluna YCM? Carotenoid C30 S-3 de 4,6 x 250mm. A quantifica??o de caroten?ides totais foi realizada por espectrofotometria UV-VIs. Todo o procedimento foi executado sob temperatura (25?C) e luz controlada. O Beta-caroteno estava presente majoritariamente, com teores de 86%, 73% e 82%, para as variedades 1, 2 e 3 e ap?s a digest?o, o perfil do Beta-caroteno passou a configurar teores de 96%, 89% e 100%, respectivamente. A efici?ncia de miceliza??o foi de 23,8%, 28% e 28,9% para as variedades 1,2 e 3, indicando a transfer?ncia do - caroteno da matriz do alimento para as micelas, correspondendo a bioacessibilidade deste composto. Esta metodologia demonstrou-se mais r?pida e mais barata, quando comparada aos estudos in vivo, que s?o mais onerosos, complexos e demandam mais tempo.

As diferentes intensidades de luz ou até mesmo a sua qualidade podem desempenhar um papel importante em algumas das principais vias metabólicas envolvidas na síntese de compostos bioativos. A radiação ultravioleta B (UV-B), além de influenciar mudanças no DNA, na atividade fotossintética e no crescimento das plantas, pode induzir a síntese e o acúmulo de metabólitos secundários. Assim, para investigar a efetividade da radiação UV-B suplementar na pós-colheita, cáqui (Diospyros kaki) e goiaba (Psidium guajava) foram submetidos a 48 horas de tratamento e posteriormente analisados em relação ao seu conteúdo de carotenoides. O acúmulo de carotenoides ocorreu de forma significativa para ambas as frutas, porém em momentos diferentes. A fim de entender os efeitos exercidos pela radiação UV-B suplementar em alimentos fontes de outros compostos bioativos, maçãs (Malus domestica) foram submetidas a 36 horas de tratamento e acompanhadas por 21 dias de armazenamento. Os parâmetros de qualidade durante o armazenamento das frutas não foram influenciados pela radiação UV-B, ocorrendo apenas perda de firmeza e de peso em todas as frutas As diferentes classes de compostos fenólicos identificados e quantificados por HPLC-MS apresentaram comportamentos diversos após o tratamento. Ácidos hidroxicinâmicos e antocianinas foram positivamente afetados pela suplementação de radiação UV-B. Para avaliar os efeitos da radiação UV-B suplementar sobre os compostos bioativos durante a pré-colheita de alimentos, alfaces verdes e roxas (Lactuca sativa) foram submetidas a 1 hora de tratamento por dia durante duas semanas. O conteúdo de carotenoides nas alfaces verdes e de compostos fenólicos nas alfaces roxas foi significativamente maior após o tratamento com radiação suplementar. Neste contexto, a radiação UV-B pode ser considerada uma tecnologia promissora no que diz respeito à modulação de compostos bioativos em alimentos, tanto durante o cultivo quanto após a colheita. / Different light intensities or even their quality may play an important role in some of the major metabolic pathways involved in the synthesis of bioactive compounds. In addition to influencing changes in DNA, photosynthetic activity and plant growth, ultraviolet B radiation (UV-B) may induce the synthesis and accumulation of secondary metabolites. Therefore, to investigate the effectiveness of post-harvest UV-B radiation, kaki (Diospyros kaki) and guava (Psidium guajava) were submitted to 48 hours of treatment and then analyzed for their carotenoid content. The accumulation of carotenoids occurred in a significant way for both fruits, but at different times. In order to understand the effects exerted by supplemental UV-B radiation on food sources of other bioactive compounds, apples (Malus domestica) were subjected to 36 hours of treatment and accompanied by 21 days of storage. The quality parameters during fruit storage were not influenced by UV-B radiation, with only loss of firmness and weight occurring in all fruits. The different classes of phenolic compounds identified and quantified by HPLC-MS showed different behavior after treatment. Hydroxycinnamic acids and anthocyanins were positively affected by the supplementation of UV-B radiation. To evaluate the effects of supplemental UV-B radiation on bioactive compounds during food cultivation, green and red lettuces (Lactuca sativa) were subjected to 1 hour of treatment per day for two weeks. The carotenoid content in green lettuce and phenolic compounds in red lettuce was significantly higher after treatment with supplementary radiation. In this context, UV-B radiation can be considered a promising technology for the modulation of bioactive compounds in food, both during and after harvest.

Resumo: O presente trabalho teve como objetivo analisar a viabilidade de uso do anidrido naftálico (NA) no tratamento de sementes de espécies gramíneas, assim como no tratamento de toletes de Saccharum spp, contra os efeitos causados por herbicidas inibidores da síntese de carotenóides. O trabalho foi constituído de quatro etapas, sendo as etapas 1 e 3 conduzidas a campo e em casa de vegetação no NUPAM/FCA/UNESP, Botucatu-SP, e as etapas 2 e 4, em estufa plástica e a campo na ESAPP e Empresa Guacho S/A, Paraguaçu Paulista-SP. Nas etapas 1, 2 e 3, de forma geral, foi estudado para as espécies Sorghum bicolor; Penisetum americanum, Panicum maximum, Brachiaria brizantha e Zea mays o efeito da combinação ou não do tratamento de sementes com NA submetidas ao uso dos herbicidas isoxaflutole (IFT) e clomazone, utilizando-se diferentes dosagens, tipos de solos e formas de aplicação. Na etapa 4 foi estudada a combinação de NA no tratamento de toletes da variedade SP 813250 (testemunhas sem NA; aplicação por imersão em solução de 3,0 g L-1 por duas horas; pulverização de 25 g ha-1 de NA sobre os colmos) e herbicida (testemunha; 112 g ha-1 de isoxaflutole; 1440 g ha-1 de metribuzin), e mais um tratamento adicional, onde os toletes foram imersos apenas em água sem adição de NA. O uso de NA (0,50% de p/p) no tratamento de sementes reduziu a intoxicação do híbrido Colorado 32 contra a ação do isoxaflutole, em pré-emergência da cultura, assim como favoreceu o aumento da produtividade dos híbridos Cargill 435 e Colorado 32, na ausência ou presença do efeito herbicida. O tratamento de sementes com NA (0,50% de p/p) protegeu a espécie S. bicolor BR 304 contra os sintomas visuais de intoxicação do isoxaflutole (112 g ha-1) e clomazone (1000 g ha-1)... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The present work had as objective to analyze the viability of naphthalic anhydride (NA) use in grass crops, through seed treatment, as well as in stalks treatment of Saccharum spp, against the effects caused by carotenoid synthesis inhibitor herbicides. The work was constituted by four step, being the 1st and 3rd ones carried out in field conditions and glasshouse at the NUPAM/FCA/UNESP, Botucatu - São Paulo State; 2nd and 4th step were conducted in plastic greenhouse and field at the ESAPP and Guacho S/A, Paraguaçu Paulista - São Paulo State. In a general way, 1st, 2nd and 3rd step was studied Sorghum bicolor; Penisetum americanum, Panicum maximum; Brachiaria brizantha and Zea mays L. species in the combination with the effect or not of seed treatment with NA submitted to the isoxaflutole and clomazone, being used different doses, kinds of soils and application forms. In 4th step, treatments included the combination of three conditions of NA stalk treatment, SP 813250 variety (check without NA; application by immersion in solution of 3,0 g L-1 of NA for two hours; spraying of 25 g ha-1 of NA over the stalks) and three herbicide conditions (check without herbicide; 112 g ha-1 at isoxaflutole; 1440 g ha-1 at metribuzin), and an additional treatment, where the stalks were just put inside water without NA. Seed treatment with NA (0,50% of w/w) reduced Colorado 32 hybrid intoxication by isoxaflutole action in preemergence of the crop, as well as favored the increase of Cargill 435 and Colorado 32 hybrids yield, in the absence or presence of herbicide effect. Seed treatment with NA (0,50% of p/p) protected S. bicolor BR 307 against the visual isoxaflutole (112 g ha-1) and clomazone (1000 g ha-1... (Complete abstract, click electronic address below) / Orientador: Edivaldo Domingues Velini / Coorientador: Sílvio José Bicudo / Banca: Dagoberto Martins / Banca: Julio Cesar Durigan / Banca: Rubem S. de Oliveira Junior / Banca: Reginaldo Teodoro de Souza / Doutor

During college, students establish nutritional habits that may last a lifetime. Fruit and vegetable consumption often decreases during the first few years of college and the vast majority of college-­‐aged students are not meeting current recommendations. Social norms theory has been shown to be an effective method for influencing behaviors in this population. This study was designed to test whether or not providing students with normative and manipulated social norms feedback could influence fruit and vegetable intakes among college-­‐aged students as evidenced by changes in skin carotenoid levels and food frequency questionnaire reports. Participants (n=244) were randomly assigned to a control group or given either normative feedback or manipulated social norms feedback regarding their skin carotenoid levels in comparison to their peers, with carotenoid levels being an objective measurement of fruit and vegetable intake. Those receiving manipulated feedback were given an artificially low carotenoid score implying that peer consumption was greater than their own. Results indicated no significant within-­‐participant changes in fruit and vegetable intakes reported in the FFQ (P=.635). While there was no change in the control groups skin carotenoid levels (P=.996), there was a borderline significant increase among those receiving normative feedback (P=.066) and a significant increase among those receiving manipulated social norms feedback (P<.001). Repeated measures of analysis showed that within-­‐participant increases in carotenoid scores were dependent on group assignment (P=.033) with an effect size of η2p=.026 which according to Cohen’s guideline is a small effect size. The distribution of carotenoid scores and FFQ results were approximately normal. Comparisons between FFQ results and skin carotenoid levels found Pearson correlation coefficients of .301 (P<.001) and other positive correlations were found between skin carotenoid levels and both exercise and BMI (.111, P=.049; -­‐.253, P<.001). The results of this study suggest that manipulated social norms feedback can increase skin carotenoid levels. The observed increases may indicate higher carotenoid containing fruit and vegetable consumption. These findings imply that social norms feedback may potentially be used as a strategy to promote and influence greater fruit and vegetable consumption among college students.

I studied carotenoids in the eggs of American coots (<i>Fulica americana</i>) from 3 study sites in Saskatchewan, Canada. I supplemented two diet types designed to reduce carotenoids in the diet of laying coots to investigate the relationship of carotenoids and the size of eggs and to examine the allocation of carotenoids into eggs.</p><p>In chapter 2, I examined influences of local environment, food quantity and food quality on egg size. Carotenoid content and stable nitrogen and carbon isotopes in yolk were measured and used to elucidate whether variation in type of food eaten contributes to egg size. By analyzing isotopes in coot tissues, I confirmed that coots use endogenous lipid reserves for egg formation but not endogenous protein reserves, and the size of eggs is more dependent on exogenous sources of nutrients. My data demonstrate that carotenoids are not causal in egg size, but are components of natural, high quality diets.</p> <p>Carotenoids are obtained through the diet and deposited into egg yolk. It has been hypothesized that concentrations and percentages of individual carotenoids can be labile and dependent on diets or maintained in an optimal balance to meet requirements of embryos. In chapter 3, I investigate deposition of carotenoids in egg yolk among nesting locales, among hens within a site and among treatments in a diet manipulation experiment. My data show maintenance in the percent composition of a suite of 3 important carotenoids, lutein, zeaxanthin and âcarotene, independent of scale of investigation and in contrast to other individual carotenoids that appear to vary in proportions based on diet. These results suggest that birds can maintain nutritional balances in their eggs despite variation in diets.</p><p>In chapter 4, I tested 3 hypotheses regarding the apportionment of carotenoids into egg yolk over the laying sequence. Without exception, concentrations of these nutrients have previously been shown to decline with egg sequence. In contrast to these findings, coots actually increased the carotenoid concentration in yolks over the laying sequence. My experimental evidence supports the explanation that this pattern of deposition depends on carotenoid availability to the laying female.

La bacteria Myxococcus xanthus, responde a la luz azul produciendo carotenoides que la protegen del daño fotooxidativo. En oscuridad la transcripción de la mayoría de los genes implicados en la síntesis de estos pigmentos es reprimida por las proteínas CarA y CarH, parálogas y funcionalmente redundantes. Ambas contienen un dominio N-terminal y otro C-terminal de unión al DNA y a cobalaminas respectivamente. Sorprendentemente, CarH, pero no CarA, depende de B12 para llevar a cabo su función represora. En este trabajo se ha demostrado que CarH y su homólogo en Thermus thermophilus (TtCarH) son fotorreceptores que utilizan la adenosilcobalmina (forma de cobalamina) como grupo cromóforo. La luz desmantela la oligomerización de estas proteínas y su unión al DNA inducidas por la adenosilcobalamina, lo que activa la expresión de los genes implicados en la carotenogénesis. Este hallazgo ha sido publicado en la prestigiosa revista PNAS (Ortiz-Guerrero et al. 2011). / The bacteria Myxocccus xanthus responds to light by producing carotenoid, protecting itself against photooxidative damage. In the dark, most of the genes involved in carotenoid synthesis are repressed by the paralogous and functionally redundant proteins CarA and CarH. Both of them contain a DNA-binding N-terminal domain and a cobalamin-binding C-terminal domain. Surprisingly, CarH, but not CarA, repressive depends on B12. In this work we showed that CarH and its homologous in Thermus thermophilus (TtCarH) are photoreceptors in which adenosylcobalamin plays the role of a chromophore. Light dismantles CarH and TtCarH adenosylcobalamine-induced oligomerization and DNA binding, activating structural genes involved in carotenoid. This finding has been reported in the prestigious journal PNAS (Ortiz-Guerrero et al. 2011)

I studied carotenoids in the eggs of American coots (<i>Fulica americana</i>) from 3 study sites in Saskatchewan, Canada. I supplemented two diet types designed to reduce carotenoids in the diet of laying coots to investigate the relationship of carotenoids and the size of eggs and to examine the allocation of carotenoids into eggs.</p><p>In chapter 2, I examined influences of local environment, food quantity and food quality on egg size. Carotenoid content and stable nitrogen and carbon isotopes in yolk were measured and used to elucidate whether variation in type of food eaten contributes to egg size. By analyzing isotopes in coot tissues, I confirmed that coots use endogenous lipid reserves for egg formation but not endogenous protein reserves, and the size of eggs is more dependent on exogenous sources of nutrients. My data demonstrate that carotenoids are not causal in egg size, but are components of natural, high quality diets.</p> <p>Carotenoids are obtained through the diet and deposited into egg yolk. It has been hypothesized that concentrations and percentages of individual carotenoids can be labile and dependent on diets or maintained in an optimal balance to meet requirements of embryos. In chapter 3, I investigate deposition of carotenoids in egg yolk among nesting locales, among hens within a site and among treatments in a diet manipulation experiment. My data show maintenance in the percent composition of a suite of 3 important carotenoids, lutein, zeaxanthin and âcarotene, independent of scale of investigation and in contrast to other individual carotenoids that appear to vary in proportions based on diet. These results suggest that birds can maintain nutritional balances in their eggs despite variation in diets.</p><p>In chapter 4, I tested 3 hypotheses regarding the apportionment of carotenoids into egg yolk over the laying sequence. Without exception, concentrations of these nutrients have previously been shown to decline with egg sequence. In contrast to these findings, coots actually increased the carotenoid concentration in yolks over the laying sequence. My experimental evidence supports the explanation that this pattern of deposition depends on carotenoid availability to the laying female.

Microbial-based industrial production has experienced a revolutionary development in the last decades as chemical industry has shifted its focus towards more sustain- able production of fuels, building blocks for materials, polymers, chemicals, etc. The strain engineering and optimization programs for industrially relevant phenotypes tackle three challenges for increased production: optimization of titer, productivity, and yield. The yield of production is function of the robustness of the microbe, generally associated with complex phenotypes. The poor understanding of complex phenotypes associated with increased production poses a challenge for the rational design of strains of more robust microbial producers. Laboratory adaptive evolution is a strain engineering technique used to provide fundamental biological insight through observation of the evolutionary process, in order to uncover molecular determinants associated with the desired phenotype. In this dissertation, the development of different methodologies to study complex phenotypes in microbial systems using laboratory adaptive evolution is described. Several limitations imposed for the nature of the technique were discussed and tackled. Three different cases were studied. Initially, the n-butanol tolerance in Escherichia coli was studied in order to illustrate the effect of clonal interference in microbial systems propagated under selective pressure of an individual stressor. The methodology called Visualizing Evolution in Real Time (VERT) was developed, to aid in mapping out the adaptive landscape of n-butanol tolerance, allowing the uncovering of divergent mechanisms of tolerance. A second case involves the study of clonal interference of microbial systems propagated under several stressors. Using VERT, Saccharomyces cerevisiae was evolved in presence of hydrolysates of lignocellulosic biomass. Isolated mutants showed differential fitness advantage to individual inhibitors present in the hydrolysates; however, some mutants exhibited increased tolerance to hydrolysates, but not to individ- ual stressors. Finally, dealing with the problem of using adaptive evolution to increase production of secondary metabolites, an evolutionary strategy was successfully designed and applied in S. cerevisiae, to increase the production of carotenoids in a short-term experiment. Molecular mechanisms for increased carotenoids production in isolates were identified.

Thesis (Ph. D.)--Ohio State University, 2007. / Title from first page of PDF file. Includes bibliographical references (p. 120-147).