Climate change and its effects on our society is a steadily growing problem. In 2010, the industry sector accounted for more than 30% of the global greenhouse gas emissions. The chemical industry is one of the industrial subsectors responsible for the highest emissions of greenhouse gas. To reach the climate goals it is therefore urgent to find more sustainable options for production of chemicals in general. Synthetic biology and microbial cell factories are growing fields that have received much attention for inferring promising sustainable alternative production routes for various compounds. When it comes to microbial cell factories, cyanobacteria infer many advantages over heterotrophs. Cyanobacteria can for instance convert atmospheric CO2 into valuable compounds through photosynthesis using the light reaction and the Calvin-Benson cycle. In the present work, the freshwater cyanobacterium Synechocystis sp. PCC 6803 is metabolically engineered to produce 1,2-propanediol; an important chemical feedstock for which there is a great interest in finding a sustainable production route as an alternative to the current petrochemical one. Seven different constructs are designed for introduction and expression of a three-step heterologous metabolic pathway for 1,2-propanediol production. Two strains of Synechocystis are successfully engineered, with the heterologous pathway chromosomally integrated at the Neutral Site I through homologous recombination with an integrative plasmid targeting this genomic site. One of the three heterologous genes (mgsA) of the pathway was successfully translated as shown in a Western immunoblot. In a SDS-PAGE a band of 40 kDa was detected, corresponding to the size of both the sADH and YqhD enzymes.

A utilização de combustíveis renováveis tem se destacado nos últimos anos, principalmente devido aos impactos ambientais gerados pelo uso de combustíveis fósseis e escassez dos mesmos. Dentre os combustíveis renováveis destaca-se o biodiesel, cujo processo produtivo apresenta como principal subproduto o glicerol bruto, que tem instigado nos últimos anos intensas discussões sobre a problemática de sua destinação e disposição final. Apesar de ser utilizado na indústria química, o seu teor de impurezas limita o seu processamento industrial. Buscando agregar valor a esse resíduo bruto e contribuir para a produção de bioenergia, este trabalho teve por intuito avaliar a produção dos biocombustíveis hidrogênio e metano, assim como de intermediários químicos a partir de glicerol bruto como substrato em reatores anaeróbios de leito fluidificado (RALF), mesofílicos (30°C), utilizando inóculo de cultura mista. O RALF-H2, com o objetivo de produzir hidrogênio e metabólitos solúveis, foi submetido à concentração de 10 g.L-1 de glicerol e variação do tempo de detenção hidráulica (TDH) de 8, 6, 4, 2, 1 e 0,5 h. Já o RALF-CH4, com intuito de produzir metano, operou sob TDH fixo de 24 h, submetido à variação da concentração de glicerol de 1, 2, 3, 4, 5 e 7 g.L-1, e portanto, da taxa de carregamento orgânico (TCO) de 1, 2, 3, 4, 5 e 7 kgDQO.m-3.d-1, respectivamente. No RALF-H2, os valores máximos de conteúdo de hidrogênio no biogás (69,2 %), produtividade volumétrica de hidrogênio (1,90 L.h-1.L-1) e rendimento de hidrogênio (0,28 molH2.mol-1glicerolconsumido) foram verificados no TDH de 0,5 h. O 1,3-propanodiol se destacou entre os metabólitos produzidos, com rendimento máximo de 0,57 mol1,3-PDO.mol-1glicerolconsumido, alcançado no TDH de 8 h. No RALF-CH4, o máximo conteúdo de metano no biogás (83,0%) foi verificado na TCO de 1 kgDQO.m-3.d-1, a maior produtividade volumétrica de metano (2,26 L.d-1.L-1) na TCO de 5 kgDQO.m-3.d-1 e o máximo rendimento (0,19 m3H4.kg-1DQOaplicada ou 0,77 molCH4.mol-1glicerolconsumido) na TCO de 4 kgDQO.m-3.d-1. Portanto, os resultados evidenciaram a possibilidade de produção de biocombustíveis e metabólitos de valor agregado utilizando glicerol bruto como substrato no processo da digestão anaeróbia. / The use of renewable fuels has been highlighted in recent years, mainly due to the environmental impacts generated by the use of fossil fuels and their scarcity. Among the renewable fuels highlights the biodiesel, whose production process displays as the main byproduct crude glycerol, which has instigated in recent years, intense debates about a problem of its destination and final disposal. Although it is used in the chemical industry, its impurities content limits its industrial processing. In order to add value to this crude residue and to contribute to the production of bioenergy, this work aimed to evaluate the production of biofuels hydrogen and methane, as well as chemical intermediates from crude glycerol as substrate in anaerobic fluidized bed reactors (AFBR), mesophilic (30°C), using mixed culture inoculum. The AFBR-H2, with the objective of producing hydrogen and soluble metabolites, was submitted to a concentration of 10 g.L-1 of glycerol and a variation of the hydraulic retention time (HRT) of 8, 6, 4, 2, 1 and 0.5 h. The AFBR-CH4, with the intention of producing methane, operated under fixed HRT of 24 h, subjected to the variation of the glycerol concentration of 1, 2, 3, 4, 5 and 7 g.L-1 and, therefore, of the organic loading rate (OLR) of 1, 2, 3, 4, 5 and 7 kgCOD.m-3d-1, respectively. In the AFBR-H2, the maximum values of hydrogen content in the biogas (69.2%), hydrogen volumetric productivity (1.90 L.h-1.L-1) and hydrogen yield (0.28 molH2.mol-1glycerolconsumed) were verified in the HRT of 0.5 h. The 1,3-propanediol was highlighted among the metabolites produced, with a maximum yield of 0.57 mol1,3-PDO.mol-1glycerolconsumed, reached in the TDH of 8 h. In the AFBR-CH4, the higher methane content in the biogas (83.0%) was detected in the OLR of 1 kgCOD.m-3.d-1, a higher volumetric productivity of methane (2.26 L.d-1.L-1) in the OLR of 5 kgCOD.m-3.d-1 and the maximum yield (0.19 m3CH4.kg-1CODapplied or 0.77 molCH4.mol-1glycerolconsumed) in the OLR of 4 kgCOD.m-3d-1. Therefore, the results evidenced the possibility of producing biofuels and value-added metabolites using crude glycerol as a substrate at the anaerobic digestion process.

A utilização de combustíveis renováveis tem se destacado nos últimos anos, principalmente devido aos impactos ambientais gerados pelo uso de combustíveis fósseis e escassez dos mesmos. Dentre os combustíveis renováveis destaca-se o biodiesel, cujo processo produtivo apresenta como principal subproduto o glicerol bruto, que tem instigado nos últimos anos intensas discussões sobre a problemática de sua destinação e disposição final. Apesar de ser utilizado na indústria química, o seu teor de impurezas limita o seu processamento industrial. Buscando agregar valor a esse resíduo bruto e contribuir para a produção de bioenergia, este trabalho teve por intuito avaliar a produção dos biocombustíveis hidrogênio e metano, assim como de intermediários químicos a partir de glicerol bruto como substrato em reatores anaeróbios de leito fluidificado (RALF), mesofílicos (30°C), utilizando inóculo de cultura mista. O RALF-H2, com o objetivo de produzir hidrogênio e metabólitos solúveis, foi submetido à concentração de 10 g.L-1 de glicerol e variação do tempo de detenção hidráulica (TDH) de 8, 6, 4, 2, 1 e 0,5 h. Já o RALF-CH4, com intuito de produzir metano, operou sob TDH fixo de 24 h, submetido à variação da concentração de glicerol de 1, 2, 3, 4, 5 e 7 g.L-1, e portanto, da taxa de carregamento orgânico (TCO) de 1, 2, 3, 4, 5 e 7 kgDQO.m-3.d-1, respectivamente. No RALF-H2, os valores máximos de conteúdo de hidrogênio no biogás (69,2 %), produtividade volumétrica de hidrogênio (1,90 L.h-1.L-1) e rendimento de hidrogênio (0,28 molH2.mol-1glicerolconsumido) foram verificados no TDH de 0,5 h. O 1,3-propanodiol se destacou entre os metabólitos produzidos, com rendimento máximo de 0,57 mol1,3-PDO.mol-1glicerolconsumido, alcançado no TDH de 8 h. No RALF-CH4, o máximo conteúdo de metano no biogás (83,0%) foi verificado na TCO de 1 kgDQO.m-3.d-1, a maior produtividade volumétrica de metano (2,26 L.d-1.L-1) na TCO de 5 kgDQO.m-3.d-1 e o máximo rendimento (0,19 m3H4.kg-1DQOaplicada ou 0,77 molCH4.mol-1glicerolconsumido) na TCO de 4 kgDQO.m-3.d-1. Portanto, os resultados evidenciaram a possibilidade de produção de biocombustíveis e metabólitos de valor agregado utilizando glicerol bruto como substrato no processo da digestão anaeróbia. / The use of renewable fuels has been highlighted in recent years, mainly due to the environmental impacts generated by the use of fossil fuels and their scarcity. Among the renewable fuels highlights the biodiesel, whose production process displays as the main byproduct crude glycerol, which has instigated in recent years, intense debates about a problem of its destination and final disposal. Although it is used in the chemical industry, its impurities content limits its industrial processing. In order to add value to this crude residue and to contribute to the production of bioenergy, this work aimed to evaluate the production of biofuels hydrogen and methane, as well as chemical intermediates from crude glycerol as substrate in anaerobic fluidized bed reactors (AFBR), mesophilic (30°C), using mixed culture inoculum. The AFBR-H2, with the objective of producing hydrogen and soluble metabolites, was submitted to a concentration of 10 g.L-1 of glycerol and a variation of the hydraulic retention time (HRT) of 8, 6, 4, 2, 1 and 0.5 h. The AFBR-CH4, with the intention of producing methane, operated under fixed HRT of 24 h, subjected to the variation of the glycerol concentration of 1, 2, 3, 4, 5 and 7 g.L-1 and, therefore, of the organic loading rate (OLR) of 1, 2, 3, 4, 5 and 7 kgCOD.m-3d-1, respectively. In the AFBR-H2, the maximum values of hydrogen content in the biogas (69.2%), hydrogen volumetric productivity (1.90 L.h-1.L-1) and hydrogen yield (0.28 molH2.mol-1glycerolconsumed) were verified in the HRT of 0.5 h. The 1,3-propanediol was highlighted among the metabolites produced, with a maximum yield of 0.57 mol1,3-PDO.mol-1glycerolconsumed, reached in the TDH of 8 h. In the AFBR-CH4, the higher methane content in the biogas (83.0%) was detected in the OLR of 1 kgCOD.m-3.d-1, a higher volumetric productivity of methane (2.26 L.d-1.L-1) in the OLR of 5 kgCOD.m-3.d-1 and the maximum yield (0.19 m3CH4.kg-1CODapplied or 0.77 molCH4.mol-1glycerolconsumed) in the OLR of 4 kgCOD.m-3d-1. Therefore, the results evidenced the possibility of producing biofuels and value-added metabolites using crude glycerol as a substrate at the anaerobic digestion process.

O 1,3-propanodiol é um composto produzido a partir da fermentação do glicerol por bactérias como a do gênero Klebsiella e que pode ser utilizado para diversas aplicações na indústria, como a produção de PTT (politrimetileno tereftalato). Esse trabalho focou na produção de 1,3-propanodiol utilizando os genes que codificam as enzimas da via retirados de uma cepa de Klebsiella previamente isolada (GLC29). Os genes foram inseridos em um plasmídeo e este em cepa hospedeira de Escherichia coli com alteração na afinidade da enzima isocitrato desidrogenase pelo seu cofator. A enzima foi modificada no genoma da bactéria e utiliza como cofator NAD em vez do NADP original. Ensaios preliminares em agitador rotativo mostraram uma melhoria de produção da cepa mutada em comparação com a Escherichia coli selvagem. Em ensaios em biorreator, variando a concentração de oxigênio dissolvido no meio, a cepa mutante apresentou uma produção 25% maior que a cepa selvagem na condição de maior produção com 5% de oxigênio dissolvido. / The 1,3-propanediol is a compound produced from glycerol fermentation by bacteria such as Klebsiella and which can be used for various applications in industry, such as the production of PTT (polytrimethylene terephthalate). This work focused on the production of 1,3-propanediol using genes encoding the enzymes of the pathway taken from a Klebsiella strain previously isolated (GLC29). The genes were inserted into a plasmid and in the host strain of Escherichia coli with changes in the affinity of the enzyme isocitrate dehydrogenase for its cofactor. The enzyme was modified in the genome of the bacteria using the cofactor NAD instead of the original NADP. Preliminary tests on shaker showed an improvement of production by the mutated strains in comparison to the wild Escherichia coli. In biorreactor cultivation, varying the concentration of dissolved oxygen in the medium, the mutant strain showed a production 25% higher than the wild type strain in the condition with 5% of dissolved oxygen.

O presente trabalho teve como objetivo avaliar o efeito de aditivos químicos, microbianos e a associação destes sobre a dinâmica fermentativa e a estabilidade aeróbia de silagens de cana-de-açúcar, além de avaliar o desempenho de vacas leiteiras de alto potencial produtivo recebendo rações com diferentes fontes de volumosos. No primeiro experimento silagens de cana-de-açúcar foram confeccionadas em silos laboratoriais de 20L, contendo válvula para escape de gases e coleta de efluente. O experimento seguiu o delineamento inteiramente ao acaso, em esquema fatorial sendo constituído por dois períodos de abertura (80 e 140 dias) com 8 tratamentos e 4 repetições. Os tratamentos avaliados foram: uréia 1%MV, 1,2- propanodiol 1%MS (1,2 p), L. L. buchneri 5x104 ufc/g (Lb), L. diolivorans em duas concentrações 1x105 ufc/g (Ld 105 e 1x10 6 ufc/g (Ld 106), L. diolivorans 1x106 ufc/g + L.buchneri 5x104 ufc/g (Ld+Lb) e 1,2-propanodiol 1%MS + L. diolivorans 1x 106 ufc/g (1,2 p+Ld 106). Após abertos os silos foram realizadas avaliações de estabilidade aeróbia em sala com ambiente controlado. Os tratamentos contendo L. diolivorans mais 1,2- propanodiol ou L. diolivorans mais L. buchneri apresentaram os menores valores de perda total de matéria seca 20,42% e 23,73%, respectivamente. Ambos tratamentos também resultaram nos menores teores de etanol, 2,1% e 3,93% da MS, e de perdas gasosas, 20,36% e 20,62%. Tanto para FDN quanto FDA, foram observados valores menores para os tratamentos contendo uréia, 1,2-p + Ld 106 Ld + Lb e Ld 105. Quanto à estabilidade aeróbia houve diferenças entre os períodos de abertura, sendo de 48,03h aos 140 dias e de 31,35h aos 80 dias. O microrganismo L. diolivorans se constituiu em uma alternativa capaz de trazer benefícios quanto às perdas fermentativas e composição bromatológica em silagens de cana-de-açúcar. O segundo experimento avaliou o desempenho de vacas recebendo rações contendo diferentes fontes de volumosos: cana-de-açúcar in natura, silagem de cana-de-açúcar inoculada com L. buchneri silagem de milho e proporções equivalentes de silagem de milho e cana-deaçúcar in natura (50:50). O experimento contou com 48 vacas Holandesas em estágio intermediário de lactação, distribuídas em múltiplos quadrados latinos 4x4 com períodos de 21 dias, sendo 14 dias de adaptação e 7 dias de coleta de amostras. Além da avaliação de desempenho, foram realizadas análises de composição bromatológica dos alimentos e do leite, e avaliações do comportamento ingestivo dos animais e da estabilidade aeróbia dos volumosos e das rações. Os dados de desempenho mostraram diferenças na ingestão de MS, com maiores valores observados para o tratamento silagem de cana-de-açúcar (23,5 kg/dia) e a mistura cana-de-açúcar mais silagem de milho (23,5 kg/dia). Todas as rações resultaram em produções elevadas de leite (22,65 kg/dia) as quais não diferiram entre si. A composição do leite variou somente para o teor de gordura, sendo maior para a silagem de milho (3,61%) e a mistura com cana (3,48%). A silagem de cana-de-açúcar mostrou-se como alternativa interessante frente à cana-de-açúcar in natura, e que ambas podem proporcionar elevadas produções desde que as rações sejam corretamente balanceadas. / The current study aimed to evaluate the effect of chemical and microbial additives and the combination over the fermentative dynamics and the aerobic stability in the sugarcane silage as well to study the performance of high producing dairy cows fed with different roughage sources. In the first trial, sugarcane silage was prepared in lab silos of 20L, with a gas relief valve and a device for collecting effluent. The experiment was randomly arranged, in a factorial design, with two openings 80 and 140 days, across 8 treatments and 4 replications. The treatments evaluated were: urea 1%, 1.2-propanediol 1%DM (1.2 p) L. buchneri 5x104 cfu/g (Lb), L. diolivorans in two concentrations 1x105 cfu/g (Ld 105) and 1x106 cfu/g (Ld 106), L. diolivorans 1x106 cfu/g plus L. buchneri 5x104 cfu/g (Ld+Lb) and 1.2- propanediol 1%DM plus L. diolivorans 1x 106 cfu/g (1,2 p+Ld 106). After opening the silos, the aerobic stability assay was performed in an environmental controlled room. The treatments containing Ld +1.2 p, or Ld 106 + Lb showed the lowest values of total DM loss, 20.42% and 23.73% respectively. Both treatments also showed the lowest levels of ethanol 2.1% and 3.93% of DM, and gases losses 20.36% and 20.62%. For both NDF and ADF, lower values were detected for the following treatments: urea, 1.2 p + Ld 106, Ld + Lb and Ld105. As for the aerobic stability, there were differences between the openings dates, 48.03h (140 days) and 31.35h (80 days). The microorganism L. diolivorans turn out to be promising alternative able to show positive results on the fermentative losses and nutritive value in sugarcane silage. The second experiment evaluated the performance of cows fed rations with different sources of roughage: fresh sugarcane, sugarcane silage, corn silage and mixture of corn silage and fresh sugarcane (50:50). Forty eight mid lactation Holstein cows were assigned to multiple latin square design 4x4, with 21-d period (14-d adaptation and 7-d the sample collection). As well the animal performance fed and milk compositions were evaluated and also ingestivo behavior of cows and the aerobic stability of forage and rations were analyzed. The performance data has shown differences in the dry matter intake (DMI), with the highest values observed for the sugarcane silage (23.5 kg/day) and the mixture (23.5 kg/day). All rations resulted in high milk production (22.65 kg/day) which did not differ among treatments. The milk composition varied only for the fat percentage, which was higher for the corn silage treatment (3.61%) and the mix (3.48%). Sugarcane silage was exhibited as an interesting forage alternative when compared to fresh sugarcane. Both options may support high milk yields by the adequacy of rations balancing.

A strategic research area today is development of polymericproducts made from renewable sources. The ways of utilizingrenewable sources studied in this thesis are using 1)prepolymers obtained by steam treatment of wood and 2) monomersobtainable by fermentation of agricultural products. Novel hemicellulose-based hydrogels were prepared by usingprepolymers obtained from steam treatment of spruce.Hemicellulose was first modified with well-defined amounts ofmethacrylic functions. Hydrogels were then prepared by radicalpolymerization with 2-hydroxyethyl methacrylate orpoly(ethylene glycol) dimethacrylate to form hydrogels. Theradical polymerization reaction was carried out in water usinga redox initiator system. The hydrogels were in generalelastic, soft and easily swollen in water. Frequency sweeptests indicated that the hydrogel system displayed prevailingsolid-like behavior. Comparison of the hemicellulose-basedhydrogels with pure poly(2-hydroxyethyl methacrylate)-basedhydrogels showed that it was possible to preparehemicellulose-based hydrogels with properties similar to thoseof pure poly(2-hydroxyethyl methacrylate)-based hydrogels. Polyester-based materials were prepared by using themonomers 1,3- propanediol and succinic acid obtainable byfermentation. α,ω-Dihydroxyterminatedoligomeric polyesters produced by the thermal polycondensationof 1,3-propanediol and succinic acid were chain-extended toobtain sufficiently high molecular weight. Depending on thechain-extension technology adopted, poly(ester carbonate)s orpoly(ester urethane)s were obtained. In the case of poly(estercarbonate)s, the chain-extended products ofα,ω-dihydroxyterminated oligomeric copolyesters werealso produced using 1,3-propanediol/1,4-cyclohexanedimethanol/succinic acid mixtures toimprove thermal and mechanical properties. Segmented poly(esterether carbonate)s fromα,ω-dihydroxyterminated oligo(propylenesuccinate)s and poly(ethylene glycol) were also synthesized toincrease the hydrophilicity. Molecular weights and polydispersity were analyzed by SECfor all materials. Their structures were also identified by NMRspectroscopy (1H NMR and 13C NMR). All characterizations werein agreement with the proposed structures. Thermal parameterswere characterized by DSC. Tensile testing anddynamic-mechanical tests were performed and in additionpreliminary processing trials were carried out in some cases.The results demonstrate the feasibility of using monomersderived from renewable sources to build up new polymericstructures endowed with a variety of physical and mechanicalproperties.

<p>A strategic research area today is development of polymericproducts made from renewable sources. The ways of utilizingrenewable sources studied in this thesis are using 1)prepolymers obtained by steam treatment of wood and 2) monomersobtainable by fermentation of agricultural products.</p><p>Novel hemicellulose-based hydrogels were prepared by usingprepolymers obtained from steam treatment of spruce.Hemicellulose was first modified with well-defined amounts ofmethacrylic functions. Hydrogels were then prepared by radicalpolymerization with 2-hydroxyethyl methacrylate orpoly(ethylene glycol) dimethacrylate to form hydrogels. Theradical polymerization reaction was carried out in water usinga redox initiator system. The hydrogels were in generalelastic, soft and easily swollen in water. Frequency sweeptests indicated that the hydrogel system displayed prevailingsolid-like behavior. Comparison of the hemicellulose-basedhydrogels with pure poly(2-hydroxyethyl methacrylate)-basedhydrogels showed that it was possible to preparehemicellulose-based hydrogels with properties similar to thoseof pure poly(2-hydroxyethyl methacrylate)-based hydrogels.</p><p>Polyester-based materials were prepared by using themonomers 1,3- propanediol and succinic acid obtainable byfermentation. α,ω-Dihydroxyterminatedoligomeric polyesters produced by the thermal polycondensationof 1,3-propanediol and succinic acid were chain-extended toobtain sufficiently high molecular weight. Depending on thechain-extension technology adopted, poly(ester carbonate)s orpoly(ester urethane)s were obtained. In the case of poly(estercarbonate)s, the chain-extended products ofα,ω-dihydroxyterminated oligomeric copolyesters werealso produced using 1,3-propanediol/1,4-cyclohexanedimethanol/succinic acid mixtures toimprove thermal and mechanical properties. Segmented poly(esterether carbonate)s fromα,ω-dihydroxyterminated oligo(propylenesuccinate)s and poly(ethylene glycol) were also synthesized toincrease the hydrophilicity.</p><p>Molecular weights and polydispersity were analyzed by SECfor all materials. Their structures were also identified by NMRspectroscopy (1H NMR and 13C NMR). All characterizations werein agreement with the proposed structures. Thermal parameterswere characterized by DSC. Tensile testing anddynamic-mechanical tests were performed and in additionpreliminary processing trials were carried out in some cases.The results demonstrate the feasibility of using monomersderived from renewable sources to build up new polymericstructures endowed with a variety of physical and mechanicalproperties.</p>

O presente trabalho teve como objetivo avaliar o efeito de aditivos químicos, microbianos e a associação destes sobre a dinâmica fermentativa e a estabilidade aeróbia de silagens de cana-de-açúcar, além de avaliar o desempenho de vacas leiteiras de alto potencial produtivo recebendo rações com diferentes fontes de volumosos. No primeiro experimento silagens de cana-de-açúcar foram confeccionadas em silos laboratoriais de 20L, contendo válvula para escape de gases e coleta de efluente. O experimento seguiu o delineamento inteiramente ao acaso, em esquema fatorial sendo constituído por dois períodos de abertura (80 e 140 dias) com 8 tratamentos e 4 repetições. Os tratamentos avaliados foram: uréia 1%MV, 1,2- propanodiol 1%MS (1,2 p), L. L. buchneri 5x104 ufc/g (Lb), L. diolivorans em duas concentrações 1x105 ufc/g (Ld 105 e 1x10 6 ufc/g (Ld 106), L. diolivorans 1x106 ufc/g + L.buchneri 5x104 ufc/g (Ld+Lb) e 1,2-propanodiol 1%MS + L. diolivorans 1x 106 ufc/g (1,2 p+Ld 106). Após abertos os silos foram realizadas avaliações de estabilidade aeróbia em sala com ambiente controlado. Os tratamentos contendo L. diolivorans mais 1,2- propanodiol ou L. diolivorans mais L. buchneri apresentaram os menores valores de perda total de matéria seca 20,42% e 23,73%, respectivamente. Ambos tratamentos também resultaram nos menores teores de etanol, 2,1% e 3,93% da MS, e de perdas gasosas, 20,36% e 20,62%. Tanto para FDN quanto FDA, foram observados valores menores para os tratamentos contendo uréia, 1,2-p + Ld 106 Ld + Lb e Ld 105. Quanto à estabilidade aeróbia houve diferenças entre os períodos de abertura, sendo de 48,03h aos 140 dias e de 31,35h aos 80 dias. O microrganismo L. diolivorans se constituiu em uma alternativa capaz de trazer benefícios quanto às perdas fermentativas e composição bromatológica em silagens de cana-de-açúcar. O segundo experimento avaliou o desempenho de vacas recebendo rações contendo diferentes fontes de volumosos: cana-de-açúcar in natura, silagem de cana-de-açúcar inoculada com L. buchneri silagem de milho e proporções equivalentes de silagem de milho e cana-deaçúcar in natura (50:50). O experimento contou com 48 vacas Holandesas em estágio intermediário de lactação, distribuídas em múltiplos quadrados latinos 4x4 com períodos de 21 dias, sendo 14 dias de adaptação e 7 dias de coleta de amostras. Além da avaliação de desempenho, foram realizadas análises de composição bromatológica dos alimentos e do leite, e avaliações do comportamento ingestivo dos animais e da estabilidade aeróbia dos volumosos e das rações. Os dados de desempenho mostraram diferenças na ingestão de MS, com maiores valores observados para o tratamento silagem de cana-de-açúcar (23,5 kg/dia) e a mistura cana-de-açúcar mais silagem de milho (23,5 kg/dia). Todas as rações resultaram em produções elevadas de leite (22,65 kg/dia) as quais não diferiram entre si. A composição do leite variou somente para o teor de gordura, sendo maior para a silagem de milho (3,61%) e a mistura com cana (3,48%). A silagem de cana-de-açúcar mostrou-se como alternativa interessante frente à cana-de-açúcar in natura, e que ambas podem proporcionar elevadas produções desde que as rações sejam corretamente balanceadas. / The current study aimed to evaluate the effect of chemical and microbial additives and the combination over the fermentative dynamics and the aerobic stability in the sugarcane silage as well to study the performance of high producing dairy cows fed with different roughage sources. In the first trial, sugarcane silage was prepared in lab silos of 20L, with a gas relief valve and a device for collecting effluent. The experiment was randomly arranged, in a factorial design, with two openings 80 and 140 days, across 8 treatments and 4 replications. The treatments evaluated were: urea 1%, 1.2-propanediol 1%DM (1.2 p) L. buchneri 5x104 cfu/g (Lb), L. diolivorans in two concentrations 1x105 cfu/g (Ld 105) and 1x106 cfu/g (Ld 106), L. diolivorans 1x106 cfu/g plus L. buchneri 5x104 cfu/g (Ld+Lb) and 1.2- propanediol 1%DM plus L. diolivorans 1x 106 cfu/g (1,2 p+Ld 106). After opening the silos, the aerobic stability assay was performed in an environmental controlled room. The treatments containing Ld +1.2 p, or Ld 106 + Lb showed the lowest values of total DM loss, 20.42% and 23.73% respectively. Both treatments also showed the lowest levels of ethanol 2.1% and 3.93% of DM, and gases losses 20.36% and 20.62%. For both NDF and ADF, lower values were detected for the following treatments: urea, 1.2 p + Ld 106, Ld + Lb and Ld105. As for the aerobic stability, there were differences between the openings dates, 48.03h (140 days) and 31.35h (80 days). The microorganism L. diolivorans turn out to be promising alternative able to show positive results on the fermentative losses and nutritive value in sugarcane silage. The second experiment evaluated the performance of cows fed rations with different sources of roughage: fresh sugarcane, sugarcane silage, corn silage and mixture of corn silage and fresh sugarcane (50:50). Forty eight mid lactation Holstein cows were assigned to multiple latin square design 4x4, with 21-d period (14-d adaptation and 7-d the sample collection). As well the animal performance fed and milk compositions were evaluated and also ingestivo behavior of cows and the aerobic stability of forage and rations were analyzed. The performance data has shown differences in the dry matter intake (DMI), with the highest values observed for the sugarcane silage (23.5 kg/day) and the mixture (23.5 kg/day). All rations resulted in high milk production (22.65 kg/day) which did not differ among treatments. The milk composition varied only for the fat percentage, which was higher for the corn silage treatment (3.61%) and the mix (3.48%). Sugarcane silage was exhibited as an interesting forage alternative when compared to fresh sugarcane. Both options may support high milk yields by the adequacy of rations balancing.

This diploma thesis is focused on the evaluation of changes in non-freezing heat transfer liquid composed of mixture of polyols and water, used in the real solar thermic system. On the base of the research a new heat transfer liquid is proposed on the platform of 1,3-polyols. The emphasis is put on the non-toxic properties of such system revealing the two chemical candidates for a new heat transfer liquid: 1,3-propanediol and glycerol. These chemicals are mixed together with water using different volume concentrations and properties of such liquids are evaluated.

This thesis deals with the evaluation of changes in properties of an antifreeze heat transfer fluid based on polyols used in real solar thermal system. Emphasis is placed on the use of substances with low impact on the environment.