Orientador: Antonio Marsaioli Junior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-03T16:21:01Z (GMT). No. of bitstreams: 1 Vilalba_FabiodeAlbuquerque_M.pdf: 570013 bytes, checksum: 8f4e3f0349848d620390519a1dc8eee4 (MD5) Previous issue date: 2003 / Resumo: O cupuaçu (Theobroma grandiflorum) é uma fruta nativa da região amazônica Brasileira cujas sementes convenientemente fermentadas, secas e torradas podem ser utilizadas para a obtenção de um produto similar ao chocolate. Uma etapa deste processo deverá também envolver, portanto, a obtenção dos nibs (cotilédones fragmentados) das amêndoas secas, separando-o da casca e do gérmen, o que pode ser feito antes ou após a torração. Esta fase do processamento da amêndoa é, ainda, realizada manualmente, de modo totalmente artesanal e pouco eficiente. O objetivo deste trabalho consistiu em pesquisar uma forma alternativa de quebra da amêndoa seca sem torrar, realizada através de um beneficiador de cilindros, seguida da utilização de um jogo de peneiras para a separação das frações. Os fatores estudados no rendimento da obtenção dos nibs e na verificação de sua pureza (percentual de casca e gérmen) foram a umidade da amêndoa inteira e a velocidade diferencial do par de cilindros de borracha do beneficiador, sendo este último a diferença das velocidades tangenciais destes cilindros. Os valores estudados das umidades foram de 5, 6 e 7%. O fator velocidade diferencial foi estudado em duas faixas, a primeira com 2,10 , 2,66 e 3,23m/s e, a segunda, com 0,84 , 1,87 e 3,23m/s. Pelos resultados obtidos, foi verificado que a variável umidade apresenta a maior influência na quebra das amêndoas. O nível de umidade de 5% da amêndoa inteira apresentou os maiores rendimentos de obtenção de nibs. A velocidade diferencial não apresentou diferença significativa nos rendimentos. Em relação aos valores dos percentuais de casca e gérmen nos nibs, nenhuma variável em estudo apresentou influência significativa nos valores desta resposta, mesmo assim todos os resultados foram bastante satisfatórios. Os percentuais de casca e gérmen encontrados no material final (nibs) foram inferior a 1,75%. O rendimento deste processo alternativo está em torno de 90% de obtenção dos cotilédones encontrados em uma determinada massa de amêndoas. Concluindo, o processo proposto constitui uma alternativa mecanizada, tecnicamente válida e superior aos métodos artesanais, apresentando bom rendimento de recuperação dos nibs, com percentuais de casca e gérmen abaixo do tolerado / Abstract: Cupuassu (Theobroma grandiflorum) is a native fruit from the Brazilian Amazon region whose seeds suitably fermented, dried and roasted can be used to manufacture a product similar to chocolate. One phase of this process must also include, therefore, the extraction of nibs (scrapped cotyledons) from the dried kernels, separating it from the shells, what can be done before or after roasting. This part of processing the kernels is still done through manual operation, in an inefficient artisan way. This work aims at devising an alternative way of cracking the dried but not roasted kernels by using a roller crusher, followed by sieving for separating the fractions. The variables studied for determining the nibs acquisition yield as well the purity of such material (expressed as percentage of shell and embryo fragments) were the moisture of the whole kernel and the differential speed of the double rollers of the crusher, this last is the difference of the tangential speeds of those rollers. The studied values of the moisture were 5, 6 e 7%. The factor differential speed was studied in two bands, the first with 2.10 , 2.66 e 3.23m/s, the second with 0.84 , 1.87 e 3.23m/s. The results confirmed that the moisture of the whole kernel was the variable that most influenced the cracking of the kernels. The moisture level of 5% of the whole kernels showed greater yield obtainment of nibs. The variation of the differential speed did not present any significant difference at yield. With regard to the percentage values of shell and embryo found in nibs, the variables considered did not show any significant difference on the response parameter, although the results were very much satisfactory. The percentage of shell and embryo found in the final material (nibs) is below the 1.75%. The yield of this alternative process is 90% of the cotyledons acquisition from certain kernels mass. In conclusion, the proposed process sets up as a mechanized alternative, of technical value and superior to the artisan methods, showing good nibs acquirement yield, and percentage of shells and embryo below the tolerated. / Mestrado / Mestre em Engenharia de Alimentos

A incidência da obesidade e do diabetes do tipo 2 tomou proporções epidêmicas nos últimos anos, atingindo bilhões de indivíduos em todo o mundo. A descoberta de formas inovadoras capazes de reduzir as alterações metabólicas associadas a estas doenças é fundamental para minimizar o seu impacto na qualidade de vida da população e na economia dos países. Muitos estudos têm demonstrado que os compostos bioativos de alimentos possuem efeitos benéficos à saúde. O camu-camu e o cupuaçu são frutas nativas da região amazônica com potencial agroeconômico ainda inexplorado, que contêm um grande número de compostos fitoquímicos que podem atuar sobre o metabolismo corporal. Desta forma, o objetivo deste estudo foi verificar os efeitos dos compostos fenólicos de extratos do camu-camu e do cupuaçu no desenvolvimento da obesidade e do diabetes tipo 2 em ensaios in vivo e in vitro, e identificar os possíveis metabólitos envolvidos nestes efeitos. Os extratos ricos em compostos fenólicos da polpa comercial destes frutos foram extraídos em fase sólida, caracterizados por cromatografia líquida de alta eficiência (CLAE/DAD) e avaliados quanto à inibição da atividade de enzimas digestivas in vitro. Os extratos obtidos foram então testados em duas diferentes concentrações (2,25 e 4,5 mg de equivalentes de catequina/Kg de peso corporal para o cupuaçu; 7 e 14 mg de equivalentes de ácido gálico/Kg de peso corporal para o camu camu) em um modelo animal de obesidade e resistência insulínica induzida por dieta com alto teor de lipídios e sacarose em camundongos C57BL/6J. Foram investigados os efeitos destes compostos sobre as homeostases glicídica e lipídica através de análises séricas, testes de tolerância à insulina e à glicose e conteúdo de lipídios hepáticos e fecais. O extrato do camu camu apresentou flavonóis, ácido elágico e elagitaninos em sua composição. A suplementação com o extrato de compostos fenólicos do camu camu reduziu o ganho de peso corporal e diminuiu a intolerância à glicose e à insulina, independente da dose administrada. No entanto, a administração destes extratos não apresentou efeitos sobre o metabolismo lipídico. Estes resultados foram associados a um possível efeito de saciedade, com consequente redução da ingestão da dieta e da glicolipotoxicidade, e com um efeito anti-inflamatório devido à diminuição dos níveis de proteína C reativa. Já o extrato de cupuaçu apresentou flavanóis, flavonas e proantocianidinas em sua composição. A suplementação com o extrato de cupuaçu na maior dose testada melhorou a homeostase da glicose e principalmente dos lipídios, protegendo o tecido hepático dos danos causados pela dieta com alto teor de lipídios e sacarose. Estes efeitos foram associados à inibição de enzimas digestivas, com consequente menor absorção de lipídios provenientes da dieta, reduzindo assim a resistência à insulina no fígado, a hiperglicemia e a dislipidemia. Ainda, foi avaliada a distribuição de metabólitos no trato gastrointestinal de camundongos após a administração aguda do extrato de cupuaçu. Foi possível identificar a complexa mistura de polifenóis presentes no extrato de cupuaçu ao longo do trato gastrointestinal, que posteriormente foi metabolizada pela microbiota. Entre os metabólitos encontrados estão as agliconas hipolaetina e isoscutelareína, e os metabolitos microbianos da epicatequina como o 3,4-diHPP-2-ol e a 5-(3,4-dihidroxiphenil)-γ-valerolactona. De acordo com estes resultados, as diferenças na composição de compostos fenólicos encontradas entre os extratos das duas frutas foram responsáveis pelos diferentes resultados nos protocolos in vivo e a identificação dos metabólitos microbianos possibilita o conhecimento dos compostos possivelmente implicados nos efeitos benéficos. Novos estudos podem contribuir para um melhor entendimento dos mecanismos, bem como quais metabólitos estão associados aos efeitos benéficos que os compostos presentes nestas duas frutas apresentaram neste estudo. / The incidence of obesity and type 2 diabetes reached epidemic proportions in recent years, arriving to billions of people around the world. The discovery of innovative ways that can reduce the metabolic abnormalities associated with these diseases is essential to minimize its impact on the population\'s quality of life and the economy of the countries. Many studies have demonstrated that food bioactive compounds have beneficial health effects. Camu-camu and cupuassu are native fruits of the Amazon region with unexplored agroeconomic potential, which contain a large number of phytochemical compounds that can act on body metabolism. Thus, the objective of this study was verify the effects of phenolic compounds of camu-camu and cupuassu extracts on the development of obesity and type 2 diabetes in vivo and in vitro, and identfy the possible metabolites involved in these effects. The phenolic compound-rich extracts were obtained from commercial frozen fruit pulps by solid phase extraction, characterized by high-performance liquid chromatography (HPLC/DAD) and evaluated for inhibition of digestive enzymes activities in vitro. Then, the extracts were tested at two different doses (2.25 and 4.5 mg catechin equivalents/kg body weight for cupuassu; 7 and 14 mg of gallic acid equivalents/kg body weight for camu camu) in an animal model (C57BL/6J mice) of obesity and insulin resistance induced by high fat high sucrose diet. The effects of extract supplementation on glucose and lipid homeostasis were assessed by serum analysis, insulin and glucose tolerance tests in mice, and contents of fat in liver and fecal samples. Camu camu extract presented flavonols, ellagic acid and ellagitannins in its composition. Supplementation with camu camu phenolic extract reduced weight gain and decreased glucose and insulin intolerance independent of the dose administered. However, no effects on lipid metabolism were found. These findings were associated with a possible effect of satiety with a consequent reduction in energy intake and glicolipotoxicity, and anti-inflammatory properties. Cupuassu extract presented flavanols, flavones and proanthocyanidins in its composition. Supplementation with cupuacu extract at the highest dose improved glucose homeostasis and plasmatic lipid levels, protecting the liver tissue from damage caused by diet rich in lipids and sucrose. These effects were associated with inhibition of digestive enzymes, with consequent lower absorption of lipids from the diet, thereby reducing the insulin resistance in the liver, the hyperglycemia and dyslipidemia. Furthermore, the distribution of metabolites in the gastrointestinal tract of mice was evaluated after acute administration of cupussu extract by HPLC-ESI-QTOF. We identified the complex mixture of polyphenols present in cupuassu extract along the gastrointestinal tract, which was subsequently metabolized by the intestinal microbiota. Among detected metabolites are hypolaetin and isoscutellarein aglycones and microbial metabolites of epicatechin as 3,4-diHPP-2-ol and 5-(3,4-dihydroxyphenyl)-γvalerolactone. According to these results, the differences in the composition of phenolic compounds found between the two fruit extracts were responsible for the different effects in vivo and identification of microbial metabolites enables the knowledge of the compounds potentially implicated in the beneficial effects. New studies can contribute to a better understanding of the metabolism and mechanisms of action.

Made available in DSpace on 2015-04-20T12:31:11Z (GMT). No. of bitstreams: 1 Rogerio Eiji Hanada.pdf: 659208 bytes, checksum: 3edd1deee7d95cb812891d0f6918caa6 (MD5) Previous issue date: 2006-03-30 / Black pod rot of cacao (Theobroma cacao) is one of the most important diseases affecting the crop worldwide. It is responsible for reducing over than 30% of the cocoa production every year. The disease is caused by Phytophthora spp. being P. palmivora (Butler) Butler one of the most common species in the cocoa growing area of Bahia, Brazil. Fungal endophytes were collected from healthy plants of cacao and cupuaçu (Theobroma grandiflorum (willd. ex Spreng.) Shum. from the states of Amazonas and Bahia, Brazil. The main objectives of this project were to isolate and to select endophytic fungi with biocontrol potential against P. palmivora. Three hundred fungal endophytes were obtained, being 104 screened against P. palmivora based on artificial inoculations of attached pods. Field experiments were performed at Almirante Cacau (Bahia-Brazil). Evaluations of disease severity were possible by using a disease rating scale specifically developed. Eight fungal isolates were selected and tested in the axenic cocoa seedlings for proving the endophytic behavior. From 103 fungi tested in the field, a Trichoderma viride Pers. ex S.F. Gray isolate was selected as the most promising candidate. This fungus was applied on cocoa plants in the field for evaluating its life span in several plant organs. The optimum spore concentration was established and formulation studies for a high performance application were evaluated. The compatibility for a tank mix application with chemical fungicides was tested and also several methods for mass production by solid state fermentation. Studies on the mode of action was carried out by evaluating the production of chitinase, b1,3 glucanase and celulase. With the exception of Tolypocladium sp., all the isolates tested: Three Fusarium, 2 Pestalotiopsis, one Curvularia and T. viride were recovered from the axenic cocoa plants. T. viride on adult cocoa plants showed an efficient epiphytic colonization on the pod surface and inside the trunk as an endophyte, been recovered after 35 and 50 days after application. The disease severity reduced by increasing the inoculum concentration up to 107 conídios.ml-1. The addition of an emulsified vegetable oil was responsible to increase the antagonist efficacy. The T. viride population over the pods decreased after 20 days. After 40 days the fungus was more dificult yor recoveretion. Disease severity increased by decreasing the antagonist population. T. viride germination was not affected by the Cooper hydroxide and Fosetil. A spore concentration of over 108 was achieved using rice as a substrate. The antagonist could grow and secrete b1,3 glucanase and chytinase in all the liquid media tested, however only on media supplemented with carboxymetilcelulose and P. palmivora mycelium were able to induce the production of celulase. / A podridão-parda do fruto de cacaueiro (Theobroma cacau) é considerada mundialmente a doença mais importante da cultura. Várias espécies de Phytophthora estão associadas à doença, sendo P. palmivora a de maior ocorrência na Bahia. O trabalho teve como objetivo isolar e selecionar fungos endofíticos com potencial de controlar a podridãoparda. A viabilização da aplicação no campo e o provável modo de ação também foram estudados. Os fungos endofíticos foram isolados de cacaueiros e cupuaçuzeiros (T. grandiflorum) dos estados do Amazonas e Bahia. A seleção foi conduzida em plantio comercial da Empresa Almirante Cacau, em Itajuípe, BA. Posteriormente, oito isolados selecionados, em campo, como os mais eficientes foram submetidos ao estudo de comprovação endofítica utilizando cacaueiros axênicos. O fungo endofítico mais eficiente no controle da doença, foi aplicado em plantas adultas e a sua sobrevivência foi avaliada nos órgãos da planta. Concentrações de 104, 105, 106, 107 e 5,0 x 107 conídios.ml-1, formulações em óleo agrícola vegetal emulsionado (Nortox) 2 % e sacarose 2%, comportamento da severidade da doença com o declínio da população do antagonista, compatibilidade com os fungicidas hidróxido de cobre e fosetil Al, produção massal de inóculo utilizando substrato de arroz e a produção extracelular de quitinase, b1,3 glucanase e celulase, em meio de cultura líquido, foram também avaliados. Foram testados 103 isolados de fungos endofíticos, sendo que Trichoderma viride foi o mais eficiente no controle da doença. Exceto Tolypocladium sp., os demais fungos: três isolados de Fusarium, dois de Pestalotiopsis, um isolado de Curvularia sp. e um de T. viride foram recuperados após a inoculação em cacaueiros axênicos. Trichoderma viride, em planta adulta, apresentou boa colonização epifítica, na parte externa do fruto, e como endofítico no interior do caule, sendo recuperado, com maior freqüência, em até 35 a 50 dias após a aplicação. A severidade da doença foi inversamente proporcional às concentrações de inóculo do antagonista, até 107 conídios.ml-1 e a adição do óleo vegetal na suspensão aumentou a eficiência do bioagente. A população de T. viride aplicado no fruto diminuiu rapidamente, em 20 dias de aplicação e, aos 40 dias, a recuperação tornou-se difícil. Por outro lado, a severidade da doença aumentou, à medida que a população de T. viride diminuiu. Trichoderma viride foi mais sensível ao hidróxido de cobre, porém a germinação dos conídios se manteve acima de 90% em solução fungicida na concentração recomenda pelo fabricante. Concentração acima de 108 conídios.g-1 do antagonista foi produzida em grãos de arroz. O bioagente cresceu e secretou b1,3 glucanase e quitinase em todos os meios de cultura, no entanto, somente os meios suplementados com carboximetilcelulose e micélio de P. palmivora foram capazes de induzir o fungo a secretar celulase.

Orientador: Nelson Horacio Pezoa Garcia / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-12T13:00:47Z (GMT). No. of bitstreams: 1 Efraim_Priscilla_D.pdf: 3005113 bytes, checksum: 644805cbb4b729e262d0a9e849b572b8 (MD5) Previous issue date: 2009 / Resumo: Este trabalho buscou contribuir com o Programa Brasileiro de Melhoramento Genético de Cacau por meio do processamento e caracterização dos principais derivados de nove clones estudados e utilizados comercialmente na Bahia por sua resistência à vassoura-de-bruxa: CEPEC 42, EET 397, TSA 654, TSA 656, TSAN 792, TSH 516, TSH 565, TSH 774, TSH 1188 e do cacau Comum, tradicional na região e suscetível à doença. Os frutos foram colhidos em um mesmo período, fermentados simultaneamente em lotes de 50 Kg e secos naturalmente ao sol. As amêndoas obtidas foram quebradas em nibs, que foram torrados em 3 diferentes tempos em forno elétrico rotativo a 150 ºC e moídos até 25 µm para obtenção dos liquors. A análise sensorial de preferência definiu o tempo de torração de cada material. A manteiga de cacau foi obtida por prensagem hidráulica dos liquors. Chocolates amargos foram produzidos pelo método convencional. A caracterização física dos frutos e sementes demonstrou diferenças entre os materiais quanto ao peso total dos frutos e rendimento de sementes com polpa por fruto. O monitoramento de temperatura e pH da massa em fermentação e a avaliação dos teores de umidade, atividade de água, pH, acidez total titulável, açúcares, ácidos orgânicos, proteínas, compostos fenólicos totais e da contagem microbiológica, durante a fermentação e ao término da secagem, demonstraram que a evolução do processo fermentativo foi distinta para cada material. A caracterização da manteiga de cacau quanto à composição em triacilgliceróis e curva de sólidos indicou que os todos os cultivares estudados, exceto o TSH 774, apresentaram melhorias em relação ao cacau Comum. A análise sensorial descritiva quantitativa dos liquors por duas equipes treinadas e de aceitação dos chocolates, por painel não treinado de consumidores, indicaram diferenças entre os materiais para alguns atributos avaliados de acordo com sua ascendência genética, sendo que os materiais descendentes dos grupos Trinitário e Forastero apresentaram maior preferência em relação aos descendentes apenas do grupo Forastero. Apesar das diferenças físicas e químicas verificadas nos liquors e chocolates, observou-se a viabilidade industrial de todos dos materiais estudados. Estudou-se ainda a influência da mistura de 0, 30, 50 e 100% de sementes danificadas pela vassoura-de-bruxa com sementes sadias na fermentação às características de amêndoas, liquors, manteiga e pó de cacau obtidos. Os perfis de temperatura e pH da massa em fermentação, os teores de açúcares e ácidos orgânicos e a análise da fração volátil demonstraram que reações que normalmente ocorrem na fermentação podem ter se iniciado nos frutos, antes da colheita. O teor de polifenóis totais das sementes danificadas foi maior que o de sementes sadias, porém, as perdas na fermentação foram maiores. Os teores de lipídios e proteínas das amêndoas e liquors foram menores quanto maior a proporção de sementes danificadas. A vassoura-de-bruxa não causou alterações nas propriedades físicas e químicas avaliadas nas manteigas de cacau. A análise da fração volátil das sementes, amêndoas e liquors indicou que o aumento do teor de sementes danificadas levou à diminuição ou ausência de importantes alquilpirazinas, indicando que o uso destas, nas porcentagens estudadas, pode prejudicar a qualidade sensorial dos derivados de cacau / Abstract: The aim of this study was to contribute with the Brazilian Genetic Improvement Program of cocoa by processing and characterizing the main cocoa-derived products from nine clones studied and used commercially in Bahia due to their resistance to ¿witches¿ broom¿: CEPEC 42, EET 397, TSA 654, TSA 656, TSAN 792, TSH 516, TSH 565, TSH 774, TSH 1188 and Comum cocoa, traditional in the area and susceptible to the disease. The fruits were harvested in the same period, fermented in 50 kg batches and sun-dried. The cocoa beans obtained were broken into nibs, roasted during 3 different times in a rotative electric oven at 150ºC and milled to 25 µm to obtain the liquors. A preference sensory evaluation defined the specific roasting time for each material. Cocoa butter was obtained by hydraulic pressing of the liquors. Bitter chocolates were produced by the conventional method. The physical characterization of the fruits and seeds demonstrated differences among the materials regarding total weight of the fruits and yield of seeds with pulp per fruit. Temperature and pH monitoring of the mass during fermentation and evaluation of moisture content, water activity, pH, total acidity, sugars, organic acids, proteins, total phenolic compounds and microbiological count during fermentation and at the end of drying demonstrated that the evolution of the fermentation process was different for each material. The triacylglycerol composition and the solid fat content of the cocoa butters from all the cultivars studied, except TSH 774, presented improvements compared to Comum cocoa. The quantitative descriptive analysis of the liquors carried out by two trained panels and the acceptance of the chocolates by an untrained consumer panel indicated differences for some of the parameters evaluated among the materials studied according to their genetic ascendance. The cultivars descending from Trinitario and Forastero groups presented greater preference in relation to the descendants of the Forastero group only. The physical and chemical characterization of the liquors and chocolates demonstrated differences among the materials studied; however, it also showed their industrial viability. The influence of the mixture of 0, 30, 50 and 100% of seeds damaged by ¿witches¿ broom¿ with healthy seeds during the fermentation process on the characteristics of cocoa beans, liquors, cocoa butter and powder was also studied. The temperature and pH profiles of the mass during fermentation, the amounts of sugars and organic acids and the analysis of volatile compounds demonstrated that some reactions that usually occur during fermentation might have been initiated in the fruits before harvesting. The total content of polyphenols in the damaged seeds was higher than in the healthy seeds; however, losses during fermentation were also greater. Lipid and protein contents of cocoa beans and liquors decreased with the increase of the proportion of damaged seeds. ¿Witches¿ broom¿ did not cause changes in the physical and chemical properties evaluated in the cocoa butters. The analysis of the volatile compounds of the seeds, beans and liquors showed that the increase of the content of damaged seeds led to the decrease or absence of the main alkylpyrazines, indicating that their use in the percentages studied may negatively affect the sensory quality of cocoa-derived products / Doutorado / Doutor em Tecnologia de Alimentos

Depuis plusieurs années, les programmes de recherche chez le cacaoyer ont mis l'accent sur l'étude des bases génétiques des caractères agronomiques d'intérêt, notamment concernant la résistance aux maladies et la qualité des fèves de cacao, qui représentent deux attributs importants pour la cacaoculture et la production de chocolat. Ce travail présente, par l'exploration du transcriptome et du génome du cacaoyer, la constitution de ressources moléculaires et l'analyse des voies de biosynthèse impliquées dans plusieurs de ces caractères agronomiques d'intérêt. L'étude du transcriptome a permis l'identification de plusieurs dizaines de milliers de gènes exprimés dans divers organes et pour différentes conditions environnementales et ont fourni de nombreux marqueurs moléculaires qui ont été utilisés pour réaliser des cartes génétiques haute densité. Les informations apportées par ce travail ont permis d’engager le séquençage du génome de la variété Criollo du cacaoyer. Son analyse et son annotation ont apporté un ensemble d'informations biologiques cruciales, depuis le catalogue des gènes et éléments mobiles jusqu'aux aspects évolutifs qui a révélé une structure du génome peu remanié par rapport à l'ancêtre commun aux dicotylédones. Par la suite, les travaux que nous avons menés pour améliorer la séquence complète ont conduit à une réduction considérable de la fragmentation chromosomique observée dans la première version. Par ailleurs 97% de la séquence assemblée et 99% des gènes sont désormais ancrés sur les chromosomes du cacaoyer. Pour commencer à exploiter cette nouvelle séquence du génome, nous avons conduit une étude QTL à partir d'une descendance entre Trinitario implantée en Guyane, permettant de localiser les régions génomiques impliquées dans la variation de la couleur des fèves de cacao. En s'appuyant sur la version améliorée du génome du Criollo, nous avons identifié deux gènes potentiellement impliqués dans la voie de biosynthèse des anthocyanines et flavonoïdes dans la principale région génomique concernée. Un des deux gènes, situé proche du marqueur situé au pic du QTL et codant pour une chalcone synthase, semble être un gène candidat prometteur. L'étude comparative de sa structure dans le génome du Criollo (à fève blanche) et du génome de l'Amelonado (à fève violette) a mis en évidence des différences structurales pouvant être à l'origine d'une modification fonctionnelle. L'ensemble des résultats présentés dans ce travail de thèse apporte une connaissance et des outils variés qui peuvent être exploités par de multiples approches intégrées pour étudier la génétique du cacaoyer. / For several years, cocoa research programs have focused on studying the genetic basis of agronomic traits of interest, especially for disease resistance and quality of cocoa beans, two important attributes for cocoa farmers and chocolate production. This work presents, by exploring the transcriptome and cocoa genome, the constitution of molecular resources and the analysis of biosynthesis pathways involved in several of these agronomical traits. The transcriptome study allowed the identification of several tens of thousands of genes expressed in various organs and for different environmental conditions and provided numerous molecular markers, used to produce high-density genetic maps. The information provided by this work led to the genome sequencing of a cocoa Criollo variety. Its analysis and annotation have provided a set of crucial biological information, from the catalog of genes and transposable elements to evolutionary aspects, which revealed a close evolutionary relationship to the eudicot putative ancestor, showing a limited number of recombination between ancestral chromosomes. Subsequently, the work we carried out to improve the complete sequence led to a considerable reduction of the chromosomal fragmentation observed in the first version. In addition, 97% of the assembled sequence and 99% of the genes are now anchored on the cocoa chromosomes. To exploit this new sequence of the genome, we conducted a QTL study from a progeny between Trinitario clones established in French Guiana, allowing to identify genomic regions involved in color trait variation observed in cocoa beans. Based on the improved version of the Criollo genome, we identified two genes potentially involved in the biosynthesis pathway of anthocyanins and flavonoids in the main genomic region concerned. One of the two genes is located nearby the QTL peak and encoding a chalcone synthase, appears to be a promising candidate gene. The comparative study of its structure in the Criollo genome (white bean) and in the Amelonado genome (purple bean) revealed several variations that could be responsible for functional modifications. The results presented in this thesis provide a variety of knowledge and tools useful to conduct multiple integrated approaches to studying cocoa tree genetics.

A incidência da obesidade e do diabetes do tipo 2 tomou proporções epidêmicas nos últimos anos, atingindo bilhões de indivíduos em todo o mundo. A descoberta de formas inovadoras capazes de reduzir as alterações metabólicas associadas a estas doenças é fundamental para minimizar o seu impacto na qualidade de vida da população e na economia dos países. Muitos estudos têm demonstrado que os compostos bioativos de alimentos possuem efeitos benéficos à saúde. O camu-camu e o cupuaçu são frutas nativas da região amazônica com potencial agroeconômico ainda inexplorado, que contêm um grande número de compostos fitoquímicos que podem atuar sobre o metabolismo corporal. Desta forma, o objetivo deste estudo foi verificar os efeitos dos compostos fenólicos de extratos do camu-camu e do cupuaçu no desenvolvimento da obesidade e do diabetes tipo 2 em ensaios in vivo e in vitro, e identificar os possíveis metabólitos envolvidos nestes efeitos. Os extratos ricos em compostos fenólicos da polpa comercial destes frutos foram extraídos em fase sólida, caracterizados por cromatografia líquida de alta eficiência (CLAE/DAD) e avaliados quanto à inibição da atividade de enzimas digestivas in vitro. Os extratos obtidos foram então testados em duas diferentes concentrações (2,25 e 4,5 mg de equivalentes de catequina/Kg de peso corporal para o cupuaçu; 7 e 14 mg de equivalentes de ácido gálico/Kg de peso corporal para o camu camu) em um modelo animal de obesidade e resistência insulínica induzida por dieta com alto teor de lipídios e sacarose em camundongos C57BL/6J. Foram investigados os efeitos destes compostos sobre as homeostases glicídica e lipídica através de análises séricas, testes de tolerância à insulina e à glicose e conteúdo de lipídios hepáticos e fecais. O extrato do camu camu apresentou flavonóis, ácido elágico e elagitaninos em sua composição. A suplementação com o extrato de compostos fenólicos do camu camu reduziu o ganho de peso corporal e diminuiu a intolerância à glicose e à insulina, independente da dose administrada. No entanto, a administração destes extratos não apresentou efeitos sobre o metabolismo lipídico. Estes resultados foram associados a um possível efeito de saciedade, com consequente redução da ingestão da dieta e da glicolipotoxicidade, e com um efeito anti-inflamatório devido à diminuição dos níveis de proteína C reativa. Já o extrato de cupuaçu apresentou flavanóis, flavonas e proantocianidinas em sua composição. A suplementação com o extrato de cupuaçu na maior dose testada melhorou a homeostase da glicose e principalmente dos lipídios, protegendo o tecido hepático dos danos causados pela dieta com alto teor de lipídios e sacarose. Estes efeitos foram associados à inibição de enzimas digestivas, com consequente menor absorção de lipídios provenientes da dieta, reduzindo assim a resistência à insulina no fígado, a hiperglicemia e a dislipidemia. Ainda, foi avaliada a distribuição de metabólitos no trato gastrointestinal de camundongos após a administração aguda do extrato de cupuaçu. Foi possível identificar a complexa mistura de polifenóis presentes no extrato de cupuaçu ao longo do trato gastrointestinal, que posteriormente foi metabolizada pela microbiota. Entre os metabólitos encontrados estão as agliconas hipolaetina e isoscutelareína, e os metabolitos microbianos da epicatequina como o 3,4-diHPP-2-ol e a 5-(3,4-dihidroxiphenil)-γ-valerolactona. De acordo com estes resultados, as diferenças na composição de compostos fenólicos encontradas entre os extratos das duas frutas foram responsáveis pelos diferentes resultados nos protocolos in vivo e a identificação dos metabólitos microbianos possibilita o conhecimento dos compostos possivelmente implicados nos efeitos benéficos. Novos estudos podem contribuir para um melhor entendimento dos mecanismos, bem como quais metabólitos estão associados aos efeitos benéficos que os compostos presentes nestas duas frutas apresentaram neste estudo. / The incidence of obesity and type 2 diabetes reached epidemic proportions in recent years, arriving to billions of people around the world. The discovery of innovative ways that can reduce the metabolic abnormalities associated with these diseases is essential to minimize its impact on the population\'s quality of life and the economy of the countries. Many studies have demonstrated that food bioactive compounds have beneficial health effects. Camu-camu and cupuassu are native fruits of the Amazon region with unexplored agroeconomic potential, which contain a large number of phytochemical compounds that can act on body metabolism. Thus, the objective of this study was verify the effects of phenolic compounds of camu-camu and cupuassu extracts on the development of obesity and type 2 diabetes in vivo and in vitro, and identfy the possible metabolites involved in these effects. The phenolic compound-rich extracts were obtained from commercial frozen fruit pulps by solid phase extraction, characterized by high-performance liquid chromatography (HPLC/DAD) and evaluated for inhibition of digestive enzymes activities in vitro. Then, the extracts were tested at two different doses (2.25 and 4.5 mg catechin equivalents/kg body weight for cupuassu; 7 and 14 mg of gallic acid equivalents/kg body weight for camu camu) in an animal model (C57BL/6J mice) of obesity and insulin resistance induced by high fat high sucrose diet. The effects of extract supplementation on glucose and lipid homeostasis were assessed by serum analysis, insulin and glucose tolerance tests in mice, and contents of fat in liver and fecal samples. Camu camu extract presented flavonols, ellagic acid and ellagitannins in its composition. Supplementation with camu camu phenolic extract reduced weight gain and decreased glucose and insulin intolerance independent of the dose administered. However, no effects on lipid metabolism were found. These findings were associated with a possible effect of satiety with a consequent reduction in energy intake and glicolipotoxicity, and anti-inflammatory properties. Cupuassu extract presented flavanols, flavones and proanthocyanidins in its composition. Supplementation with cupuacu extract at the highest dose improved glucose homeostasis and plasmatic lipid levels, protecting the liver tissue from damage caused by diet rich in lipids and sucrose. These effects were associated with inhibition of digestive enzymes, with consequent lower absorption of lipids from the diet, thereby reducing the insulin resistance in the liver, the hyperglycemia and dyslipidemia. Furthermore, the distribution of metabolites in the gastrointestinal tract of mice was evaluated after acute administration of cupussu extract by HPLC-ESI-QTOF. We identified the complex mixture of polyphenols present in cupuassu extract along the gastrointestinal tract, which was subsequently metabolized by the intestinal microbiota. Among detected metabolites are hypolaetin and isoscutellarein aglycones and microbial metabolites of epicatechin as 3,4-diHPP-2-ol and 5-(3,4-dihydroxyphenyl)-γvalerolactone. According to these results, the differences in the composition of phenolic compounds found between the two fruit extracts were responsible for the different effects in vivo and identification of microbial metabolites enables the knowledge of the compounds potentially implicated in the beneficial effects. New studies can contribute to a better understanding of the metabolism and mechanisms of action.

<p dir="ltr">Macambo (<i>Theobroma bicolor</i>) is a relative of cacao that was identified as an underutilized crop in San Martín, Peru with potential for valorization. Current processing for macambo seeds results in waste of two-thirds of the fruit, so the two waste streams (husk and pulp-shell) were characterized by proximal analysis to identify useable components. From the pulp-shell waste, soluble polysaccharides and proteins were extracted. The resulting hydrocolloid formed physical gels that had predominantly solid-like elastic behavior and can partially recover after being deformed by a constant stress. Macambo pulp-shell waste has potential for extracting functional ingredients for thickening, forming gels, retaining water, and stabilizing emulsions.</p><p dir="ltr">One third of food in the United States is never eaten, and waste occurs throughout the food system. Produce is a significant portion of food waste, but shelf life can be extended by optimizing postharvest practices. Conducting precooling in-field at point of harvest with mobile units was proposed by researchers in the field of food waste reduction as an adaptable method to reduce food waste and was evaluated in this study for innovation and barriers to use. Based on expert interviews and a literature review, it was concluded that in-field precooling units are hindered by infrastructure requirements and effective precooling is already optimized at large scale with centralized cooling activities. Flexible cooling technology could be part of a solution for very small growers in the United States and internationally in developing economies, where access to precooling and full cold chain is still lacking.</p>

No description available.

Le cacaoyer (Theobroma cacao, L.) est cultivé dans les tropiques humides d'Amérique latine, d'Afrique et d'Asie. Principale source de revenus d'une agriculture essentiellement familiale, les systèmes agroforestiers à base de cacaoyers fournissent de nombreux services écologiques et contribuent à la conservation de la biodiversité. Ces systèmes associent plantes ligneuses et cultures de manière simultanée, dans une gamme importante et peu décrite de richesses et de structures botaniques. Les connaissances actuelles sur leur productivité se limitent à des essais en milieu contrôlé associant un petit nombre d'espèces. Nos travaux testent l'hypothèse d'une relation de compromis entre le niveau de productivité des cacaoyers et le niveau de biodiversité hébergée par les agroforêts à base de cacaoyers. Nous caractérisons ces compromis sur un gradient de situations de production paysannes, reposant sur la structure verticale, l'intensité de gestion de la composante végétale et le contexte bio-physique des agro-systèmes étudiés. Sur un dispositif de 36 cacaoyères paysannes situées dans la région de Talamanca, Costa Rica, nous montrons que des variations significatives de la structure végétale reflètent les modes de gestion de la parcelle et affectent le rendement par cacaoyer (295 à 667g/arbre/an) mais pas le rendement en cacao (136 kg/ha/an), ni le volume végétal global (400 m3/ha). Ces variations de la structure végétale affectent peu la diversité α des peuplements de plantes associées, d'épiphytes, d'amphibiens, de reptiles, de mamifères, d'invertébrés du sol et de la litière, mais leur diversité β répond de manière contrastée à ces variations d'habitat. Les relations de compromis que nous mettons finalement en évidence entre la productivité en cacao marchand à l'hectare ou à l'arbre et les niveaux de biodiversité atteints sont de nature (linéaire, cubique, quadratique) et de tendances (négative, postive) contrastées selon le taxon considéré et remettent en cause les résultats de certains travaux récents. Nos travaux dévoilent également des situations de compromis optimales, offrant des perspectives positives pour l'intensification écologique des systèmes agroforestiers tropicaux. / Cacao (Theobroma cacao, L.) is cultivated in the humid tropics of Latin America, Africa and Asia. Cocoa-based agroforestry systems are the main source of income for smallholders' families and provide numerous ecological services and contribute to biodiversity conservation. These systems associate trees with crops simultaneously, in a wide and poorly described range of botanical and structural plant diversity. The current knowledge about their productivity is restricted to controlled trials with a low numer of associated plant species. Our work tests the hypothesis of a trade-off relationship between the level of cocoa productivity and the level of biodiversity hosted in cocoa-based agroforests. We characterize these trade-offs on a gradient of production situations, based on the vertical structure, the management intensity of the vegetal component and the bio-physical context of cocoa-based agroforests. On the base of a 36 on farm cocoa plots network located in Talamanca, Costa Rica, we show that significant variations in the vegetation vertical structure reflect farmer's management strategies and affect the cocoa yield per tree (295 to 667g/tree/year) but neither the cocoa yield per plot (136 kg/ha/year), nor the global plant volume (400 m3/ha). The variations in vegetation structure poorly affected the α-diversity of 7 plant and animal taxa, but their β-diversity gave contrasted and significant responses to habitat variations. Finally, the trade-offs relationships we display between dry cocoa productivity per hectare or per tree and the observed biodiversity levels showed contrasted forms (cubic, quadratic, linear) and tendances (positive, negative) according to the taxa considered and question results from recent publications. Our work also revealed optimal trade-off situations which offer positive prospects for the ecological intensification of tropical agroforestry systems.

La cacaoculture est aujourd'hui considérée comme un des principaux facteurs de déforestation en milieu tropical. Dans de nombreux pays, elle repose en effet sur un modèle technique peu durable de monoculture intensive impliquant le déplacement des zones de production. Souvent délaissés par la recherche en raison de leurs faibles rendements en cacao marchand, les systèmes agroforestiers à base de cacaoyer apparaissent aujourd'hui comme une alternative crédible pour faire face aux nouveaux enjeux de la cacaoculture mondiale. Au Centre-Sud du Cameroun, la cacaoculture repose en grande partie sur des cacaoyères agroforestières anciennes, ce qui pose l'hypothèse qu'un modèle de cacaoculture durable basé sur des systèmes de culture agroforestiers est possible. Notre objectif est de mieux comprendre le fonctionnement de ces systèmes agroforestiers à base de cacaoyer, à travers leurs caractéristiques, leurs évolutions et leurs déterminants, ainsi que les conditions de leur production en cacao marchand sur le long terme. Notre démarche articule à la fois une évaluation des systèmes agroforestiers complexes à l'échelle de la parcelle et une analyse compréhensive des pratiques techniques sur le temps long. La thèse repose sur un dispositif de recherche mis en place dans la région du Centre, dans trois zones de culture différenciables par leurs conditions pédo-climatiques (Bokito, Zima et Ngomedzap). Une enquête d'agriculteurs a été réalisée à grande échelle (1 171 exploitations regroupant 1 638 cacaoyères), complétée par des observations spécifiques réalisées dans un réseau de 61 cacaoyères exploitées par 40 agriculteurs. Résultats. (1) Nos résultats d'enquête, complétés par des observations spécifiques, ont confirmé la durabilité des systèmes agroforestiers à base de cacaoyer aux plans agro-écologique et socio-économique. (2) Dans la zone de transition forêt-savane, considérée comme sub-optimale pour la cacaoculture, nous avons montré que le modèle technique adopté par les agriculteurs leur permet d'installer sur savane des cacaoyères viables sur le long terme. (3) Une méthode participative de quantification de la valeur d'usage des espèces par les agriculteurs, appliquée sur le réseau d'observation, a confirmé la multifonctionnalité des systèmes agroforestiers et mesuré la place qu'y occupe la composante cacaoyère. (4) L'évaluation des peuplements cacaoyers, réalisée en adaptant la méthode de diagnostic agronomique régional aux systèmes complexes, a permis d'identifier les facteurs limitant le rendement en cacao marchand. Celui-ci s'avère étroitement lié à la structure des peuplements cacaoyers et à celle des peuplements associés. (5) Enfin, l'analyse compréhensive des pratiques des agriculteurs sur le long terme, en lien avec l'évolution technique des peuplements cacaoyers a confirmé la flexibilité des systèmes agroforestiers à base de cacaoyer. Cette flexibilité peut être traduite par des trajectoires d'évolution des pratiques et des trajectoires de structures spécifiques. Ce travail fournit une nouvelle base méthodologique pour l'analyse et l'évaluation des systèmes agroforestiers complexes. L'ensemble des résultats et des connaissances produites dans ce travail permet de formuler des propositions pour la mise au point d'un nouveau modèle technique en cacaoculture. / Cocoa cultivation is generally considered as one of the main deforestation factors in the Tropics. Indeed, in many countries it is based on a low-sustainability technical model of intensive monocrops with shifting production zones. Cocoa agroforests, which are often neglected by research due to their low fermented dried cocoa yields, now appear to be a credible alternative for taking up the new challenges facing world cocoa production. In Centre-South Cameroon, cocoa cultivation is largely based on old cocoa agroforests, which suggests that a sustainable cocoa growing model based on agroforestry systems is possible. Our aim was to more effectively understand how these cocoa agroforests function, through their characteristics, their evolution and their determinants, along with the conditions for their long-term fermented dried cocoa production. Our approach consisted of an evaluation of cocoa agroforests on a plot scale and a comprehensive analysis of technical practices over the long term. The thesis is based on an experimental design set up in the Centre region, in three growing zones that could be differentiated through their pedo climatic conditions (Bokito, Zima and Ngomedzap). A large-scale survey was undertaken (1,171 farms grouping 1,638 cocoa plantations) completed by specific observations collected in a network of 61 cocoa agroforests managed by 40 farmers. Results. (1) Our survey results, completed by specific observations, confirmed the sustainability of the cocoa agroforestry systems from an agro ecological and socio-economic viewpoint. (2) In the forest-savannah transition zone, which is considered sub-optimum for cocoa cultivation, we confirmed that the technical model adopted by the farmers enabled them to set up cocoa agroforests on savannah that were viable over the long term. (3) In the network, a participatory method applied to quantify the use value given to the species by the farmers confirmed the multi-functionality of cocoa agroforests and measured the place occupied in it by cocoa. (4) The evaluation of the cocoa tree stands, achieved by adapting the regional agronomic diagnosis method to complex systems, revealed the factors limiting cocoa yield. It was proved that cocoa yield is closely linked to the structure of the cocoa tree stand and that of the intercropped stands. (5) Lastly, a comprehensive analysis of farmer practices over the long term, in association with the evolution of the technical management of the cocoa tree stands, confirmed the flexibility of cocoa agroforestry systems. This flexibility was reflected in the evolution pathways for specific practices and structures. This work provides a new methodological basis to assess the performance of complex agroforestry systems. All the results and knowledge produced by this work make it possible to draw up proposals for developing a new technical model for cocoa growing.