Orientadores: Antonio José de Almeida Meirelles, Roberta Ceriani / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-22T08:04:00Z (GMT). No. of bitstreams: 1 Silva_SimoneMonteiroe_D.pdf: 2753093 bytes, checksum: 5edb0c2d58e280b4aa2179e93036df00 (MD5) Previous issue date: 2013 / Resumo: Os óleos vegetais são importantes componentes da dieta humana e devem ser refinados antes do consumo. A demanda dos consumidores por produtos mais saudáveis, assim como regulamentações ambientais cada vez mais rígidas têm forçado os processadores de óleos vegetais a buscarem mudanças e aperfeiçoamento dos processos. Neste contexto, este trabalho de tese tem como objetivo investigar e aperfeiçoar o refino físico de óleos vegetais, com ênfase na etapa de branqueamento. Numa primeira etapa, uma metodologia para quantificar simultaneamente carotenos e tocoferóis foi desenvolvida e validada, e posteriormente, utilizada pelo grupo de pesquisa. Então, a etapa de branqueamento de óleo de palma, atualmente o óleo mais consumido mundialmente, foi estudada sob diferentes aspectos: (1) determinação da cinética, equilíbrio e parâmetros termodinâmicos do processo de adsorção de carotenos e fósforo em terra clarificante acidamente ativada; (2) influência de diferentes procedimentos na cor do óleo de palma refinado; (3) influência do tipo de terra clarificante na cor do óleo de palma refinado. Estes estudos foram importantes para um melhor entendimento do processo de branqueamento de óleo de palma, e algumas conclusões foram obtidas: a adsorção de carotenos e fósforos pela terra clarificante acidamente ativada ocorre por via química, e é um processo endotérmico; a utilização de novos procedimentos na etapa de branqueamento do óleo de palma pode melhorar a coloração obtida ao final do processo; o refino em duas etapas utilizando duas desodorizações em condições moderadas de temperatura apresentou melhor coloração final quando comparado ao processo utilizando a mesma quantidade de terra clarificante e tempo de desodorização; uma hipótese foi sugerida para explicar como o tipo de terra clarificante (neutra ou acidamente ativada) pode interferir na coloração final do óleo de palma. Ainda são necessários mais estudos para otimizar a etapa de branqueamento e os novos procedimentos sugeridos. Por último, a etapa de desacidificação por via física foi estudada através de simulação computacional e dados experimentais disponíveis na literatura. Foram comparadas duas abordagens matemáticas: a diferencial e a flash. Esta última apresentou resultados mais realísticos quanto aos perfis de acidez e perda de óleo neutro. Foi ainda considerada na abordagem flash equações de transferência de calor. Dessa forma, pode-se concluir que este trabalho de tese apresentou avanços nos processos de refino físico de óleos vegetais para obtenção de produtos de maior qualidade final e menor consumo de insumos / Abstract: Vegetable oils are important compounds of the human diet and they should be refined before consumption. Consumers demand for healthier products as well as stiff environmental legislation are forcing refining industries towards changes and improvement of processes. In this context, this thesis has as main objective to investigate/improve the physical refining of vegetable oils, emphasizing the bleaching step. As first step, a HPLC methodology for simultaneous quantification of carotenes and tocols was developed and validated, and lately, it was used by our research groups. Then, bleaching step of palm oil, nowadays the most consumed oil in the world, was studied under different aspects: (1) determining kinetics, equilibrium and thermodynamic parameters of adsorptive removal of carotenes and phosphorus onto acid activated bleaching earth; (2) influence of different procedures on final color of palm oil; (3) influence of bleaching earth kind on final color of palm oil. These studies were important for a better understanding of bleaching process of palm oil, and some conclusions were obtained: adsorptive removal of carotenes and phosphorus onto acid activated bleaching earth occurs by chemisorption and it is endothermic; new procedures in the bleaching step can improve final color of palm oil when using the same amount of bleaching earth and deodorization time; a hypothesis was proposed to explain how the kind of bleaching earth can interfere in the final color of palm oil. Further studies are still necessary to optimize bleaching step and the new procedures suggested. Later, physical deacidification was studied by computer simulation and experimental data from literature. It was compared two mathematical approaches: differential and flash distillations. This last one presented better results regarding acidity and neutral oil loss profiles. In this approach, it was considered the heat transfer equations. In this way, this thesis presents an advance in refining process towards high quality products and less consumption of inputs / Doutorado / Engenharia de Alimentos / Doutora em Engenharia de Alimentos

Os chás destacam-se como a segunda bebida mais consumida no mundo. Além disso, seu consumo tem se modernizado, adequando-se aos anseios dos consumidores. São uma boa fonte de compostos com atividade antioxidante, os quais podem trazer benefícios à saúde humana pelo retardamento do processo de envelhecimento, assim como pela prevenção de doenças crônicas, tais como o câncer e doenças cardiovasculares. Infusões de ervas processadas e frescas foram estudadas quanto a teor de compostos fenólicos totais, perfil de flavonóides e atividade antioxidante. Os teores de compostos fenólicos totais, determinados pelo método de Folin–Ciocalteu, variaram de zero a 46,46 mg/g. Os flavonóides miricetina, quercetina e campferol foram identificados e quantificados por cromatografia líquida de alta eficiência (CLAE) nos chás preto, verde e de camomila. A atividade antioxidante foi avaliada pelo método do seqüestro do radical 2,2'-difenil-1-picrilhidrazil (DPPH) e pelo sistema modelo β-caroteno/ácido linoléico (BCAL), usando hidroxitolueno butilado (BHT) e α- tocoferol como padrões. As maiores atividades encontradas pelo sistema modelo BCAL foram para as infusões de chá preto, mate, capim-limão, camomila e funcho, enquanto as infusões de ervas frescas apresentaram as menores atividades. Entretanto, quando a atividade antioxidante foi avaliada pelo método do DPPH, as infusões de ervas frescas de menta e funcho foram as que apresentaram as maiores atividades. Os chás processados que apresentaram os menores valores de IC50 foram verde e preto (147,63 µg/mL e 288,60 µg/mL, respectivamente). Os resultados obtidos demonstraram que as infusões analisadas são boas fontes de compostos com atividade antioxidante. / Teas are the second most consumed beverage in the world. Besides this, their consumption has been modernized to suit the consumers? desires. They are a good source of compounds presenting antioxidant activity, which can bring benefits to human health by retarding the process of aging, as well as by preventing chronic diseases, such as cancer and cardiovascular diseases. Processed teas and herbal infusions were studied for their total polyphenol content, flavonoid profile, and antioxidant activity. Total phenolics, determined by Folin?Ciocalteu procedure, ranged from zero to 46.46 mg/g. Flavonoids myricetin, quercetin, and kaempferol were identified and quantified by High Efficiency Liquid Chromatography (HPLC) in black, green, and chamomile tea. Antioxidant activity was evaluated using 2,2 Diphenyl-1-picryl-hydrazyl method (DPPH) and β-carotene bleaching test (BCB), with butylated hydroxytoluene (BHT) and α-tocopherol as reference substances. The highest activities using BCB were found for infusions of black tea, maté, lemongrass, chamomile, and fennel, whereas fresh herbal infusions presented the lowest activities. However, when the antioxidant activity was evaluated by DPPH, fresh herbal infusions of mint and fennel presented the highest activities. Processed teas with the lowest IC50 values were green and black (147.63 µg/mL and 288.60g/mL and 288.60 µg/mL, respectively). The results obtained demonstrate that the infusions analyzed are good sources of compounds presenting antioxidant activity.

O objetivo geral deste trabalho foi elaborar formulados alternativos, à base de mistura de alimentos convencionais, provenientes da cesta básica distribuída por entidades do município de Piracicaba-SP, entre outros alimentos, com o intuito de suplementar a alimentação enteral de idosos. Os alimentos utilizados para a elaboração dos formulados foram fubá, óleo de soja, arroz, feijão, macarrão, sardinha, leite em pó, amido de milho, ovos, proteína texturizada de soja, couve- manteiga e cenoura. Foram realizadas análises físicas: gotejamento e viscosidade; análises químicas: composição centesimal, digestibilidade de proteína, determinação do mineral ferro, diálise de ferro in vitro, determinação de -caroteno e fatores antinutricionais, assim como fenólicos totais e ácido fítico; análises microbiológicas: Salmonella, Bacillus cereus, Estafilococos coagulase positiva e Coliformes totais; análise de custos dos formulados e elaboração do manual de boas práticas de manipulação. Os resultados obtidos foram submetidos às análises estatísticas, empregando-se o teste de Tukey, utilizando o software SAS (1996) e análises de regressão, pelo programa Microsoft Office Excel (2003). Concluiuse, com esse estudo, que os formulados líquidos apresentaram os melhores resultados nutricionais, físicos, microbiológicos, e a maior viabilidade econômica, sendo, por isso, sugeridos para a suplementação enteral de idosos. Dentre eles, pode-se eleger uma entre duas alternativas, conforme a necessidade clinica do idoso, considerando-se todos os aspectos nutricionais e físicos dos formulados. O formulado F3 destacou-se nas seguintes características: disponibilidade de ferro, quantidades de -caroteno, digestibilidade, viscosidade e gotejamento. Já o formulado F4 teve destaque na distribuição do valor calórico total de macronutrientes, apresentando valores ideais para os idosos, segundo as recomendações internacionais, além do maior valor calórico entre os formulados e boa viscosidade e gotejamento. No que diz respeito ao valor calórico total, os formulados apresentaram de 131 a 216 calorias por 250ml, os tratamentos líquidos apresentaram valor calórico superior ao dos formulados em pó. Pode-se dizer que os formulados contribuem com percentual médio de 20% de adequação, em relação às recomendações de macronutrientes sugeridas pela DRI (2002) para idosos, quando ingeridas duas porções de 250mL/dia de formulados líquidos ou em pó. / The aim of this study was to develop alternative formulas for elderly enteral feeding using conventional foods from food staples distributed in the city of Piracicaba-SP. The foods used in the formulas were: corn meal, soya oil, rice, beans, pasta, sardine, powder milk, corn starch, eggs, texturized soya protein, cabbage, and carrot. Physical analyses (dripping and viscosity), chemical analyses (centesimal composition, protein digestibility, iron determination, iron dialyses in vitro, -carotene determination, anti-nutritional factors - tannins and phytate), microbiological analyses (Salmonella, Bacillus cereus, positive Staphylococcus coagulase and total Coliforms), cost analyses and elaboration of a manual for good manipulation practices were carried out. Results were analyzed statistically using Tukey test, SAS software (1996) and for regression using Microsoft Office Excel (2003). Liquid formulas presented better nutritional, physical and microbiological results as well as economical viability Therefore were suggested for elderly enteral nourishment. According to the elderly clinical needs, two alternatives are suggested among liquid formulas. The F3 formula is noticeable for iron availability, quantity of -carotene, digestibility, dripping and viscosity. The F4 formulas is noticeable for the total calorie distribution of macronutrients as well as for having a higher number of calories and good viscosity and dripping which is ideal for the elderly, according to international recommendations. The formulas presented a range of 131 to 216 calories/250ml. Liquid formulas presented higher number of calories than powder formulas. The formulas contributed with a 20% adequacy regarding to macronutrient recommendations for the elderly from DRI (2002), when two servings of 250mL.day-1 (powder or liquid) were ingested.

A anemia causada por deficiência de ferro é a forma mais comum de todas as anemias e afeta grande parcela da população mundial, especialmente o grupo materno-infantil: lactentes, pré-escolares, escolares, gestantes e nutrizes. A carência acarreta atraso no desenvolvimento psicomotor e cognitivo de crianças menores de dois anos, além de diminuição da eficiência da função imune, e agravamento de doenças infecciosas. O presente estudo teve por objetivos avaliar a disponibilidade de ferro em alimentos fontes deste e de vitamina A, ovo (o), cenoura (c) e couve (cv), bem como verificar a interação do ferro com α e ß-caroteno, e com outros interferentes. As misturas foram de ovo com cenoura e ovo com couve em diferentes proporções (1:1), (1:1,5), (1:2), (1:2,5) e (1:3) e também dos 3 alimentos sozinhos, totalizando 13 amostras. As amostras, após pré-preparo e cocção, foram submetidas às análises de composição centesimal, ferro, taninos, ácido fítico, ácido oxálico, α- e β- caroteno e ferro dialisável. Os dados foram submetidos à análise estatística empregando o teste de Tukey, regressão polinomial e correlação, utilizando o software SAS (1996). Os valores para umidade variaram de 83,93 a 95,74 g/100g, apresentando diferenças significativas entre as distintas amostras, assim como para proteínas (0,55 a 7,22g /100g) e extrato etéreo (0,21a 5,94 g/100g). Foi observado que em relação à umidade, extrato etéreo, proteínas, cinzas e β-caroteno houve correlação significativa para o ferro dialisável, sendo que para umidade e β-caroteno essa correlação foi positiva e para os demais componentes foi negativa. Os teores para taninos, fitatos e oxalatos variaram de 0,78 a 1,40 mEqcatequina/g, 0,17 a 0,42 mg/g, e 0,06 a 0,30% respectivamente. A amostra 2 (1o:1,5c) apresentou teores mais elevados de taninos (1,40 mEqcatequina/g), e as amostras 6 (1o:1cv) e 7 (1o:1,5cv) apresentaram teores mais elevados de oxalato e teores de ferro dialisável mais baixo, em relação às outras misturas. O teor de α-caroteno para as amostras contendo cenoura variou de 2.435,27 a 4.609,83 µg/100g. Quanto ao β-caroteno observou-se variações de 2.406,75 a 7.881,55 µg/100g, sendo evidenciado valores mais elevados para as amostras com cenoura. Numericamente, as misturas com maior proporção de cenoura e couve apresentaram os maiores resultados para o ferro dialisável, que variou entre 0,99 a 7,43%, sendo que a amostra 5 (1o:3c) apresentou o melhor resultado, seguida pelas amostras 1, 2, 3, 4, 8, 9 e 10. Pelos resultados obtidos pode-se concluir que as misturas com maior proporção de cenoura e couve aumentaram a disponibilidade do ferro, principalmente pelo maior conteúdo de ß-caroteno. Não houve interferência significativa dos antinutricionais sobre o ferro dialisável, embora os taninos e ácido oxálico atuaram como fatores limitantes quando observados os valores numéricos nas amostras com maiores concentrações de cenoura e couve. O ß-caroteno apresentou correlação positiva significativa com o ferro dialisável, exercendo influência na redução do efeito dos antinutricionais, especialmente dos fitatos, na disponibilidade do ferro, principalmente nas amostras com cenoura. Assim, pode ser recomendada a utilização de misturas de alimentos como ovo e cenoura, sendo que este último deve estar em maiores proporções em refeições destinadas a populações carentes em ferro. / The anemia caused by iron deficiency, is the form more common of all the anemias and it affects large portion of the world population, especially the maternal-infantile group: infants, child-school, children, pregnant woman and nutrizes, carting delay in the development psicomotor and cognitive of smaller children two years old, besides decrease of the efficiency of the immune function, and worsening of infectious diseases. This research aims to estimate the iron availability in food iron sources and vitamin A, egg (e), carrot (c) and cabbage (cb), as well as to verify the interaction of the iron with α, ß-carotene and with other interferents, being mixed the foods in study, resulted 13 food mixtures in different proportions (1:1), (1:1.5), (1:2), (1:2.5) and (1:3) include three food alone. The food mixtures, after prepare and cooking, was submitted the analyses of centesimal composition, levels of iron, tannin, phytic acid, oxalic acid, α, ßcarotene and in vitro iron dialysability. The statistical analysis was determinate by SAS System include polynomial regression and correlation. The values for moisture varied from 83.93 to 95.74 g/100g, showing significant differences among the different mixtures, as well as for proteins (0.55 to 7.22 g/100g) and fat (0.21 to 5.94 g/100g). There was significant correlation with regard to moisture, fat, protein, ash and ß-carotene to in vitro iron dialysability, being positive correlation to moisture and ß-carotene, and negative to other components. The levels of tannins, phytate and oxalates ranged 0.78 to 1.40 mEqcatequina/g, 0.17 to 0.42mg/g and 0.06 to 0.30%, respectively. The mixture 2 (1e:1.5c) showed high levels of tannins (1.40mEqcatequina/g), and the mixtures 6 (1e:1cb) and 7 (1e:1.5cb) high levels of oxalates, and lower levels of in vitro dialysability iron, in relation to the other mixtures. The values for α- carotene to mixtures with carrot ranged 2,435.27 to 4,609.83 µg/100g. With regard to ß-carotene, ranged variations to 2,406.75 from 7,881.55 µg/100g, being observed higher levels for the mixtures with carrot. The numbers showed that the mixtures with larger carrot and cabbage proportion showed the bigger results for the in vitro dialysability iron, ranges to 0.99 from 7.43 %, and the sample 5 (1e:3cb) it presented the bigger value, proceeded by the mixtures 1, 2, 3, 4, 8, 9 and 10. It’s concluded that the mixtures with larger carrot and cabbage proportion increased the iron availability, especially for the largest content of ß-carotene. There was not significant interference of antinutritional in the in vitro iron dialysability, although tannins and oxalates were an inhibitor when observed the numeric values in the mixtures with larger carrot and cabbage concentrations. The ß-carotene showed significant positive correlation with in vitro iron dialysability, increasing influence in the reduction of the antinutritional factors effect, especially phytates, in the iron availability, mainly in the mixtures with carrot. Therefore, the use of food mixtures as egg and carrot can be recommended, and this last one should be in larger proportions in meals destined to lacking populations in iron.

Les crises énergétiques, en particulier celles liées aux combustibles fossiles, conduisent à une utilisation accrue des nouvelles énergies renouvelables. De plus, les recherches visent également à utiliser des matériaux de stockage de l'énergie qui sont plus efficace, tels que les batteries, et à utiliser des matériaux économes en énergie. Une OLED (diode électroluminecente organique)) est une diode émettrice la lumière (LED) qui sont connues grâce leur potentiel élevé dans les applications d'affichage, de signalisation et d'éclairage. Parmi ces matériaux organiques, les caroténoïdes constituent une classe importante de molécules linéaires conjuguées qui présentent un degré de délocalisation électronique élevé et de dynamique ultra-rapide. Le colorant joue un rôle important dans le fonctionnement des cellules solaires sensibilisées par colorant (CSSCs) ou d'autres matériaux bi fonctionnels. Dans cette recherche, nous avons étudié la photo stabilité des caroténoïdes (-carotène et fucoxanthine) à l'aide du kaolin modifié et de l'antioxydant. La photostabilité du -carotène et de la fucoxanthine peut être améliorée par la kaolinite modifiée et l'antioxydant. La fabrication OLED utilisant la fucoxanthine n'est pas suffisante à cause de la bande interdite de la fucoxanthine (Fx) qui est très grande et peut être un isolant. NPD (50 nm) / Fx (1 nm) / Alq3 (85 nm) a un EQE de 0,12% et un CIE (0,416 ; 0,5302). Les dispositifs OLEDs en utilisant le ß-carotene comme couche de transport du trou et la curcumine comme couche émettrice ont donné une couleur jaune avec un EQE de 0,02%. / Energy crises, especially fossil fuel-based energy, lead to increased use of new energy and renewable energy. In addition, research is also directed towards more efficient energy use through the development of energy storage materials such as batteries, and the use of energy-efficient materials. An OLED (organic light-emitting diode) is a light-emitting diode (LED) and is known for their high potential in a display, signage, and lighting applications. Carotenoids constitute an important class of linear -conjugated molecules that exhibit a high degree of electronic delocalization and ultrafast dynamic. The dye that is used as a photosensitizer plays an important role in the operation of DSSCs or other bifunctional material. In this research, we studied the photostability of the carotenoid compound (-carotene and fucoxanthin) using natural antioxidant and modified kaolinite. In this work, the carotenoid stability by the carotenoid compound/antioxidant binary is studied for the first time. Photostability of -carotene can be enhanced by antioxidant and modified kaolinite. Photoprotection efficiency of -carotene by curcumin was higher than fucoxanthin. Modified kaolinite decreased photodegradation of - carotene by shielded and protected from direct UV irradiation. Antioxidant fucoxanthin and curcumin decreased the electrochemical gap of the binary material. The electrochemical gap of carotene/curcumin is -1.61 eV, carotene/fucoxanthin is -1.75 eV compare to the only -carotene - 2.04 eV. Fucoxanthin can keep the first oxidation stage of -carotene. Therefore at the binary compound, electron was still reversible, but not for curcumin. Fucoxanthin in OLED devices NPD/Fx/ETL reduced significanly the EQE (%) almost 80% in Alq3 and 76% in BAlq. OLED device: NPD(50nm)/Fx(1nm)/Alq3(85nm) yielded an effeciency quantum external yield, EQE = 0.12% and CIE (0.4160, 0.5302). Fabrication OLED using -carotene as HTL and curcumin as EL layer resulted yellow color , EQE = 0.02%, with the composition MoO3(15nm)/NPD(40nm)/Car(10nm)/ Cur(15nm)/Alq3(70nm)/Ca(100nm). For application of - carotene in OLED device, its perfomance can be improved by using curcumin at the fabrication.

Carotenoids are natural pigments that have antioxidation and antimutagenic abilities. They are produced with the help of new technological methods. For example, carotenoid yeast Rhodotorula glutinis produces -carotene with the yield of up to 6 – 10 mg/g of the dry substance. The method of the mass spectrometry with the nanoelectrospray in the positive mode was optimized for the determination of -carotene, lycopene and astaxanthin in this project. Ionizing voltage of 4 kV and the sample flow rate of 15 – 80 nl/min through the spray silica fused capillary with the internal diameter of 25 µm were found to be the optimum parameters of the analysis. A mixture of chloroform with the addition of ammonia was used as a spray solvent for both standard and cellular samples. During the process of ionization by nanoelectrospray, -carotene and lycopene form cation radical [M] • + with the molecular mass to charge ratio (m/z) of 536, while asthaxanthin forms the protonated molecule [M + H]+ with the m/z of 597. The partial lysis of individual Rhodotorula glutinis cells was demonstrated under microscope in the organic solvents tetrahydrofuran and dimethylsulfoxide. Chloroform, acetone, acetonitrille, methanol and isopropanol did not affect the cells after a 15 min treatment.

The purpose of this research was to study vitamin A status in grazing horses throughout the year and to evaluate the effectiveness of vitamin A and β-carotene as supplements. Vitamin A status was assessed by serum retinol concentrations (SR) and the relative dose response (RDR) which was adapted for use in the horse. The horses (45 Thoroughbred mares) were divided into three diet groups: pasture and hay only (PH); pasture, hay and vitamin A-free concentrate (PHC); and hay and vitamin A-free concentrate (HC). The mares, as well as their foals, were assessed for vitamin A status during the summer, fall and winter. After eight months, each diet group was subdivided and supplemented with either: retinyl palmitate at two times the recommended level (A), the equivalent in water dispersible β-carotene (B), or a placebo (C). Supplementation continued for 20 months during which the vitamin A status was assessed every 60 days in the mares, and at birth in the neonates. During both the depletion and the repletion phase the mares were kept on a regular breeding schedule and the reproductive rates were determined as well as the general health of the mares and their offspring. The RDR proved more sensitive at detecting changes in vitamin A status than SR, and a combination of both was used. A measurable decline in vitamin A stores was seen in the HC group within 2 months, and in PH and PHC groups during the winter. The HC group remained lower in vitamin A status throughout the study. A seasonal fluctuation of vitamin A status was observed regardless of supplementation. Supplementation with retinyl palmitate improved vitamin A status in all three diet groups, however, supplementation with β-carotene did not. Both neonates and young growing horses were lower in vitamin A status than the adult. A respiratory infection observed in the weanlings affected vitamin A status as well. Supplementation of the dam had no effect on neonatal vitamin A status. Deleterious effects on reproductive rates and health were also observed with vitamin A depletion. Supplementation of β-carotene had a negative effect on reproductive rates in this study. / Ph. D.

Tesis por compendio / A pesar de los enormes beneficios potenciales que ofrecen los cítricos genéticamente modificados (GM), su liberación en campo suscita preocupaciones acerca de su potencial impacto ambiental y posibilidad de que muestren efectos deletéreos inesperados desde un punto de vista agronómico. Las principales preocupaciones que plantea el uso de la transformación genética para la mejora de este cultivo de vida larga, propagación vegetativa y compleja biología reproductiva son: (1) la transferencia de los transgenes vía polen a variedades compatibles de especies de Citrus y afines; (2) la estabilidad de los transgenes a largo plazo; (3) la aparición de efectos pleiotrópicos adversos derivados de la integración y la expresión de los transgenes sobre las principales características agronómicas y fenotípicas del cultivo. Todas estas cuestiones han sido ampliamente estudiadas en otros cultivos anuales GM que ya son o no comerciales. Sin embargo, puesto que el empleo de la transformación genética en la mejora de árboles frutales todavía se encuentra en sus inicios, actualmente se dispone de muy poca información al respecto para estos cultivos. Por todo ello, el futuro de los arboles transgénicos en el ámbito comercial permanece aún incierto, aunque actualmente se dispone de la tecnología para producirlos. Por otro lado, en el caso concreto de los cítricos, no existen variedades transgénicas comerciales ni evidencias inequívocas de que esta herramienta sea realmente útil para afrontar con éxito objetivos de mejora concretos. Lograr cumplir objetivos de mejora tan importantes como la mejora de la calidad nutri-funcional de los frutos cítricos mediante ingeniería genética podría contribuir a una mayor aceptación de esta tecnología por parte del público, puesto que se trata de una mejora dirigida primeramente al consumidor. En este trabajo nos hemos planteado afrontar parte de los aspectos que en gran medida limitan la aceptación y comercialización de cítricos GM, mediante (1) la realización de un experimento de campo con cítricos GM para evaluar su seguridad ambiental y la ausencia de efectos agronómicos adversos (2) el abordaje de un objetivo de mejora de la calidad nutri-funcional de la naranja concreto mediante ingeniería metabólica con la finalidad de reforzar sus propiedades saludables. El experimento de campo consistió en una plantación de cítricos transgénicos que portaban únicamente los genes marcadores uidA y nptII cuya finalidad fue estudiar la viabilidad de la transformación genética en la mejora de genotipos cítricos comercialmente importantes. Este huerto experimental nos sirvió para estimar la frecuencia máxima de dispersión de los transgenes por polen bajo condiciones de polinización abierta y estudiar los factores ambientales, genéticos y fenológicos que la determinan, para así poder proponer medidas de contención apropiadas en futuras plantaciones de cítricos GM. También sirvió como primera aproximación para abordar cuestiones básicas como el estudio de la estabilidad de la expresión de los transgenes a largo plazo (tras 7 años de establecimiento en campo) bajo condiciones reales de cultivo y su potencial impacto sobre la morfología, fenología y calidad de la fruta de los cítricos transgénicos. Los estudios realizados, aunque no resuelven todas las preocupaciones concernientes a los cítricos GM, aportan información crucial relativa a su seguridad y comportamiento en campo, inexistente hasta el momento, que puede servir como base para futuros ensayos de campo con cítricos GM y como guía para las políticas de regulación de su plantación (caso-a-caso). Por otro lado, en este trabajo se ha logrado desarrollar una estrategia para inducir producción temprana de fruta e incrementar el contenido de b-caroteno (pro-vitamina A, con elevada capacidad antioxidante) en la pulpa de una variedad de naranjo dulce mediante ingeniería metabólica. Dicha estrategia consistió en el silenciamiento mediado por RNAi del gen de una β-caroteno hidroxilasa de naranjo (CsβCHX), implicada en la conversión de b-caroteno en xantofilas, combinado con la sobreexpresión del gen FLOWERING LOCUS T de naranjo (CsFT) en plantas transgénicas juveniles de naranjo dulce cv Pineapple. Posteriores ensayos con el animal modelo Caenorhabditis elegans demostraron que la naranjas enriquecidas ejercían un efecto antioxidante in vivo un 20% mayor que las naranjas control isogénicas. Este es el primer ejemplo exitoso de ingeniería metabólica para incrementar el contenido de β-caroteno (o cualquier fitonutriente) en naranjas y demuestra el potencial que tiene la ingeniería genética para el enriquecimiento nutricional de cultivos frutales leñosos. / Pons Bayarri, EA. (2014). Towards application of genetic engineering in citriculture: 1) assessing dispersal, long-term stability and phenotypic impact of transgenes in citrus trees and 2) improving nutri-functional quality of orange fruit through metabolic engineering [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/43588 / Compendio

Le β-carotène (Bc) est un caroténoïde connu pour son activité pro vitaminique A. La consommation de fruits et légumes riches en Bc est donc particulièrement encouragée, principalement dans les pays en développement. Cependant, au cours d'un repas, la biodisponibilité du Bc est influencée par la présence des macro et microconstituants des aliments. L'objectif général de ce travail a été d'étudier les interactions alimentaires pouvant survenir au cours des différentes étapes du processus d'absorption du Bc. La bioaccessibilité du Bc, évaluée à l'aide d'un modèle de digestion in vitro, a révélé que la quantité de Bc micellarisé des aliments de base (patate douce orange (PDO) et banane plantain) des pays du Sud se retrouve augmentée dans certaines préparations tratitionnelles. De plus, dans une simple association de deux aliments, nos résultats ont montré que la naringine de jus d'agrumes diminuait la bioaccessibilité du Bc de la PDO car ce flavanone se micellarise et entre en compétition avec le Bc. L'effet de flavanones sur l'absorption intestinale du Bc a ensuite été étudié à l'aide d'un modèle cellulaire de type Caco-2. L'ensemble des glycosides de flavanones testés, mais principalement l'hespéridine (Hes), ont augmenté l'absorption du Bc. L'expérimentation in vivo chez la gerbille n'a quant à elle pas montré d'effet de l'Hes sur la bioefficacité du Bc de la PDO. Par contre, nos données suggèrent que l'Hes, dans le cas d'un régime carencé en caroténoïdes et vitamine A, pourrait stimuler l'activité de la β,β-carotène mono-oxygénase par un mécanisme impliquant le facteur de transcription PPARγ. / Β-carotene (Bc) is a carotenoid mainly known for its provitaminic A activity. Therefore, the consumption of fruits and vegetables rich in Bc are promoted, especially in developing countries. However, during a meal, the Bc bioavailability was modulated by other macro or micronutrients from food. The main objective of this study was to evaluate food interaction occurring during the different steps of Bc absorption. The Bc bioaccessibility, evaluated through an in vitro digestion, showed that the micellarization of Bc staple foods from South countries (orange fleshed sweet potato (OFSP) and plantain) increases in some traditional food preparations. Moreover, we observed that the Bc bioaccessibility from OFSP decreases in the presence of Citrus juice because of the naringin which competes for incorporation into mixed micelles. Effects of flavanones were then assessed on intestinal Bc uptake using Caco-2 cells. Among flavanone glycosides tested, mainly hesperidin (Hes), increased Bc uptake. In vivo experimentation with gerbil did not shown any effect of Hes on the bioefficacity of Bc from OFSP. By contrast, our data suggest that under a low Bc or vitamin A free diet, Hes might enhance BCMO1 activity through its action as agonist of PPARγ.

Plusieurs acteurs moléculaires impliqués dans les variations de qualité de la viande ont été récemment mis en évidence chez le poulet. Ma thèse a pour objectif d’approfondir l’étude de leur régulation en étudiant l’impact de facteurs alimentaires en interaction avec l’origine génétique des animaux. Il s’est articulé autour de deux thématiques qui impliquent des acteurs moléculaires et des critères de qualité de viande indépendants : le rôle de l’AMPK (AMP-activated protein kinase) dans le contrôle du turnover du glycogène musculaire et des caractères qui en dépendent (pH, rétention d’eau, couleur) et l’implication de BCMO1 (β, β-carotene-15,15’-monooxygenase) dans les variations de teneurs en pigments caroténoïdes et de coloration. Nos résultats soulignent dans les deux cas la possibilité de moduler les caractères de qualité via l’alimentation avec des réponses qui dépendent des caractéristiques génétiques des animaux. Nos travaux ont aussi permis d’améliorer la compréhension de la régulation des biomarqueurs étudiés par les nutriments et la génétique et contribueront à terme à la mise en place de nouvelles stratégies de production permettant d’optimiser la qualité du poulet de chair en réponse aux attentes de la filière et des consommateurs. / Several molecular mechanisms involved in the variations of poultry meat quality were recently identified in chickens. My thesis aims to further study their regulation by exploring the impact of dietary factors in interaction with the genetic origin of animals. It was structured around two themes that involve independent molecular mechanisms and meat quality criteria: the role of AMPK (AMP-activated protein kinase) in the control of muscle glycogen turnover and related meat traits (pH, water retention, color), and the involvement of BCMO1 (β, β-carotene-15, 15'-monooxygenase) in controlling levels of carotenoid pigments and yellow color. Our results emphasize in both cases the possibility of modulating quality traits through nutrition, with effects that depend on the genetic characteristics of animals. Our work has also improved the understanding of the regulation of studied biomarkers by genetics and nutrients. This should contribute to the development of new production strategies to optimize the quality of broilers in response to expectations of poultry producers and consumers.