To mitigate the problems of depleting and soaring price of fossil fuels, the production and use of renewable energy have been vigorously promoted. In Europe, the role of biologically-derived fuels and in particular biodiesel is gradually increasing in prominent. Rapeseed biodiesel is the most widely produced in Europe. As a consequence, enormous amount of by-products from production processes are being generated. Current strategies for managing these by-products (mainly rapeseed meal and crude glycerol) seem not to be economically sustainable. More efficient utilisation could add more value to the production chain which in turn would raise the competitiveness of biodiesel compared to petro-diesel. The aim of the project reported in this thesis was to study the feasibility of producing a value added product, polyhydroxybutyrate (PHB), from by-products generated from rapeseed biodiesel production processes as well as to investigate the effects of methanol, a major impurity in crude glycerol, on growth of Cupriavidus necator, a PHB-producing micro-organism.The preliminary study of C. necator growth in crude glycerol based media revealed that optimum concentration of crude glycerol was in a range 15-25 g/L. It was also found that slight changes in the carbon to nitrogen ratio of the feedstock did not significantly affect the growth while methanol at concentrations beyond 10 g/L did. A model based on a saturation equation was developed and used to successfully predict the inhibition of growth by methanol. From the developed model, mechanisms of the inhibition were proposed. The model could also be used to predict satisfactorily growth or productivity rates in other systems containing short-chain alcohols. The growth in solutions derived from rapeseed meal (designated as hydrolysate) via solid-state fermentation by Aspergillus oryzae followed by hydrolysis of the fermented solids was also studied. The biomass production was found to increase as a function of initial free amino nitrogen (FAN) concentration presented in the hydrolysate. However, at higher initial FAN concentrations, a lower conversion of nitrogen to biomass was observed. PHB production was studied using a feedstock which was a mixture of the hydrolysate and crude glycerol. Total biomass concentration reached 28.8 g/L at 120 h with 86% PHB content. PHB productivity and PHB yield on glycerol were 0.21 g/L•h and 0.32 g/g respectively. These results were comparable with those obtained when pure glycerol and synthetic crude glycerol were used, suggesting that, technically, the use of the generic rapeseed- and crude glycerol-based feedstock to produce PHB is feasible.Overall, the feasibility of producing PHB from rapeseed biodiesel by-products has been demonstrated. The satisfactory result leads to the more important outlook that the generic feedstock derived from rapeseed biodiesel by-products has the potential to be used to produce a wide range of products depending on the micro-organism used. Further development of this process to improve nutrient production efficiency as well as product yields and subsequent integration of the process into the biodiesel production process could well be an important contribution in the development of a sustainable biodiesel industry.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-08-09T17:41:31Z No. of bitstreams: 2 Tese - Patricia Antonio - 2016.pdf: 2346033 bytes, checksum: 5d733f10f72d4427f3e205bc5f8674ad (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-10T11:36:49Z (GMT) No. of bitstreams: 2 Tese - Patricia Antonio - 2016.pdf: 2346033 bytes, checksum: 5d733f10f72d4427f3e205bc5f8674ad (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-10T11:36:49Z (GMT). No. of bitstreams: 2 Tese - Patricia Antonio - 2016.pdf: 2346033 bytes, checksum: 5d733f10f72d4427f3e205bc5f8674ad (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-03-04 / The animal production potential grows fast and food production is always being challenged to meet this need in the nutrition as well as in the economic context. Forage plants are the basis of the diet of ruminants in most production systems in tropical regions. Forage availability varies during the year depending on weather conditions. It turns necessary to have stored forage to supplement the animals. Silage is the most widely used form of stored forage. The tropical grasses ensiling is an alternative to traditional silage crops because it can be used the surplus produced in the rainy season. However, the fermentation process can be altered by many factors, making difficult the production of good quality silage so there is the need of using additives as a means of improving the nutritional and fermentative quality of these types of silages. Thus, this study aimed to use moisture sequestering additives: soybean, canola, sunflower and cotton meal, in the production of B. brizantha silage in order to improve the nutritional value, fermentative value, degradability and digestibility of these silages, thus obtaining information about alternative materials for the production of good quality silage. The experiment was carried out a part in experimental PVC silos and estimated the bromatologic parameters, effluent and gas losses, N-NH3, pH, titratable acidity, protein fractions and silage degradability, and the other part in experimental silos with a capacity of 200 kg to evaluate the performance of sheep fed with these silages through intake and total tract digestibility of nutrients, metabolic and rumen parameters of these animals. The silages of Brachiaria brizantha with sequestering moisture additives were effective in keeping concentrations of plasma metabolic and ruminal profile parameters within the reference levels, increasing the readily soluble A fraction and decreasing the C protein fraction consequently improving the ruminal degradability, and reducing the silage losses. Therefore it is recommended its use as an alternative to ruminant nutrition. / O potencial de produção animal cresce num ritmo acelerado e a produção de alimentos está sempre sendo desafiada a suprir essa necessidade tanto no contexto nutricional quanto econômico. As plantas forrageiras constituem a base da dieta dos ruminantes na grande maioria dos sistemas de produção das regiões tropicais. A disponibilidade de forragem é variável durante o ano, dependendo de condições climáticas. Isto gera a necessidade de se ter forragem conservada sendo a silagem a forma mais utilizada. A ensilagem de capins tropicais é uma alternativa à ensilagem de culturas tradicionais podendo ser utilizada o excedente produzido na época das águas. No entanto, o processo fermentativo pode ser alterado por alguns fatores, dificultando a confecção de silagens de boa qualidade, havendo assim a necessidade do uso de aditivos como meio de melhora da qualidade nutricional e fermentativa desses tipos de silagens. Dessa forma, o presente trabalho visou utilizar aditivos sequestrantes de umidade: farelos de soja, canola, girassol e algodão, na confecção de silagens de Brachiaria brizantha com o objetivo de melhorar o valor nutritivo, valor fermentativo, degradabilidade e digestibilidade dessas silagens, obtendo assim, informações sobre materiais alternativos para a confecção de silagens de qualidade. O experimento foi realizado uma parte em silos experimentais de PVC e estimados os parâmetros bromatológicos, as perdas por efluentes, perdas por gases, N-NH3, pH, acidez titulável, frações proteicas e a degradabilidade da massa ensilada, e outra parte em silos experimentais com capacidade para 200 kg para avaliar o desempenho de ovinos alimentados com essas silagens, através de consumo e digestibilidade total dos nutrientes, parâmetros metabólicos e ruminais destes animais. As silagens de Brachiaria brizantha com aditivos sequestrantes de umidade foram eficientes em manter as concentrações dos metabólicos plasmáticos e parâmetros de perfil ruminal dentro dos níveis de referência, aumentar a fração A prontamente solúvel e diminuir a fração C da proteína consequentemente melhorando a degradabilidade ruminal, e diminuindo as perdas da silagem. Portanto sendo recomendado o seu uso como alternativa para a nutrição de ruminantes.

La fermentation en milieu solide (FMS) est un procédé biotechnologique particulièrement bien adapté au traitement de la biomasse végétale. Cette technologie, extrapolable à l’échelle industrielle, peut répondre aux besoins actuels du marché de l’alimentation animale en développant des produits fermentés visant à améliorer les qualités des matières premières d’un point de vue nutritionnel et fonctionnel. Les travaux de recherche présentés dans cette thèse développent ces aspects à partir du tourteau de colza issu du procédé de trituration des graines.Un premier criblage a mis en évidence le potentiel de certaines souches fongiques en prenant en compte les performances de croissance et leur capacité à enrichir le tourteau en protéines totales et digestibles. Des teneurs comparables à celles du tourteau de soja, principale source de protéine en nutrition animale, ont d’ailleurs été atteintes. Une activité biologique d’intérêt a ensuite été démontrée in vitro sur cellules immunitaires. Cette voie, inexplorée jusqu’à présent à partir de culture en milieu solide, confère une valeur ajoutée au produit fermenté via un apport en composés immunomodulateurs naturels d’origine fongique comme les β-glucanes. Ces derniers suscitent un intérêt croissant car ils représentent une alternative à l’utilisation des antibiotiques comme facteur de croissance. L’étude consacrée à la phase d’optimisation du procédé FMS a permis d’accroître les performances de croissance de la souche A. sojae et de définir une stratégie visant à maintenir les conditions optimales de croissance en réacteur pré-pilote à couche profonde. Des complications liées à la granulométrie très fine du tourteau de colza et à la qualité microbiologique qui évolue en cours de fermentation dans des conditions non stériles ont pu être identifiées mais le procédé mis en place a alors permis de les maîtriser en partie. / Solid state fermentation (SSF) is a biotechnological process particularly well adapted to the treatment of vegetable biomass. This technology adjustable to industrial scale, is adapted to current challenges in animal nutrition by developing fermented products in order to improve the quality of raw materials from a nutritional and functional point of view. The work presented in this thesis considers these issues using rapeseed meal from the oil crushing industry.A first screening study highlighted several fungal strains based on their growth performance and protein enrichment (total and digestible) of the substrate. Levels similar to those of soybean meal, the main protein source in animal nutrition, have also been reached. An interesting biological activity was then reported in vitro on immune cells. To date, this way has never been investigated concerning solid state culture. The presence of fungal immunomodulatory compounds as β-glucan confers added value to the fermented product. Recently, there has been growing interest in natural immunomodulators as they represent an alternative to the use of antibiotics for growth promotion in food animal production. SSF process optimization improved the growth performance of the strain A. sojae and helped to determine the best strategy to maintain optimum growth conditions at larger scale in the deep-bed pre-pilot reactor. Complications related to the fine particle size of the rapeseed meal and the changes in microbiological quality during fermentation under non-sterile conditions were also identified and partially controlled by the process.

The aim of this study was to investigate and compare the climate impact from different arable land use options and protein feeds aimed for cattle. This has been made by executing two life cycle assessments (LCAs). The first LCA aimed to compare the following three arable land use options: Cultivation of wheat used for production of bioethanol, carbon dioxide and DDGS Cultivation of rapeseed used for production of RME, rapeseed meal and glycerine Fallow in the form of long-term grassland The second LCA aimed to compare the three protein feeds DDGS, rapeseed meal and soybean meal. In the LCA of arable land, the functional unit 1 ha arable land during one year was used and the LCA had a cradle-to-grave perspective. The LCA of protein feeds had the functional unit 100 kg digestible crude protein and had a cradle-to-gate perspective, hence the use and disposal phases of the feeds were excluded. Bioethanol, DDGS and carbon dioxide produced at Lantmännen Agroetanol, Norrköping, were investigated in this study. The production of RME, rapeseed meal and glycerine were considered to occur at a large-scale plant in Östergötland, but no site-specific data was used. Instead, general data of Swedish production was used in the assessment. The wheat and rapeseed cultivations were considered to take place at the same Swedish field as the fallow takes place. The protein feed DDGS was produced at Lantmännen Agroetanol and the rapeseed meal was assumed to be produced at a general large-scale plant in Sweden. In the soybean meal scenario, a general case for the Brazilian state Mato Grosso was assumed and no specific production site was investigated. Data required for the LCAs was retrieved from literature, the LCI database Ecoinvent and from Lantmännen Agroetanol. In the LCA of arable land use options, system expansion was used on all products produced to be able to compare the wheat and rapeseed scenarios with the fallow scenario. In the LCA of protein feeds, system expansion was used on co-products. The products in the arable land use options and the co-products in the protein feed scenarios are considered to replace the production and use of products on the market with the same function. The result shows that the best arable land use option from a climate change perspective is to cultivate wheat and produce bioethanol, carbon dioxide and DDGS. This is since wheat cultivation has a higher yield per hectare compared to rapeseed and therefore a bigger amount of fossil products and feed ingredients can be substituted. To have the arable land in fallow is the worst option from a climate change perspective, since no products are produced that can substitute alternative products. Furthermore, the result shows that DDGS and rapeseed meal are to prefer before soybean meal from a climate change perspective, since soybean meal has a higher climate impact than DDGS and rapeseed meal. This can be explained by the smaller share of co-products produced in the soybean meal scenario compared to the DDGS and rapeseed meal scenarios. Since the production and use of co-products leads to avoided greenhouse gas emissions (since they substitute alternatives), the amount of co-products being produced is an important factor. A sensitivity analysis was also executed testing different system boundaries and variables critical for the result in both LCAs. The conclusion of this study is that arable land should be used to cultivate wheat in order to reduce the total climate impact from arable land. Furthermore, it is favorable for the climate if DDGS or rapeseed meal are used as protein feeds instead of imported soybean meal.