<p dir="ltr">The periparturient period involves coordinated physiological adaptations as the dairy cow transitions from a non-lactating to lactating state. The ability of dairy cattle to adapt to the onset of lactation is impacted both by physiological and nutritional factors, and a poor transition can result in reduced productivity and welfare for the animal. Additionally, disease and disorder development are heightened in the transition period, with increased risk for involuntary culling occurring in early lactation. This study aimed to evaluate if the amount of late gestational muscle reserves and prepartum supplementation of branched-chain volatile fatty acids (BCVFA) impacts health and production parameters in multiparous, periparturient dairy cattle. Forty-eight multiparous Holstein dairy cattle were assigned to either a high or low muscle group (HM or LM, respectively) based on their <i>longissimus dorsi</i><i> </i>muscle depth 42 days before expected (BEC). After assignment to group, cattle were then randomly assigned to a control (CON; 80 g/d soyhull pellets as-fed basis) or BCVFA (40 g/d isobutyrate product, 20 g/d isovalerate product, 20 g/d 2-methybutyrate product, fed as calcium salt products on an as-fed basis) treatment, which was top-dressed in the prepartum period only. After parturition, treatment was no longer provided and cattle were fed a common lactating diet. Blood samples, ultrasound images, and feed intake were collected and recorded from 42 BEC through 28 days in milk (DIM), milk yield and composition data was collected from parturition until 28 DIM.</p><p dir="ltr">HM cattle began mobilizing muscle reserves prior to parturition, while LM cattle began to accrete muscle reserves prior to parturition. This difference in prepartum muscle utilization did not impact other body measurements (i.e. body weight or body condition score) between the groups of cows but did result in increased blood glucose concentrations prepartum for HM cows compared with LM. This increase in glucose concentrations is likely due to the increased supply of gluconeogenic precursors as a result of the degradation of muscle tissue. The difference in glucose concentration was not observed postpartum, neither was there a difference in tissue mobilization between the groups postpartum. HM cattle had greater DMI both pre- and postpartum, and produced greater yields of milk, milk fat, milk protein, and milk lactose postpartum compared to the LM group. Despite the increased milk yield, there was no difference in feed efficiency between the groups, as the HM cows consumed more feed. Prepartum supplementation of BCVFA did not impact body measurement changes throughout the entire transition period, but did increase pre- and postpartum DMI, likely due to increased fiber digestibility. The BCVFA treatment increased blood glucose concentrations both pre- and postpartum and reduced milk urea nitrogen concentrations postpartum, likely due to improved nitrogen efficiency. Results show that prepartum supplementation of BCVFA has an improved ruminal carryover effect into early lactation.</p><p dir="ltr">At 14 days BEC and 7 DIM, an intravenous glucose tolerance test (IVGTT) was performed on a sub-set of cows, to evaluate if insulin response could be a mechanism impacting the efficiency and production differences observed between muscle groups and BCVFA supplementation. BCVFA supplementation increased glucose area under the curve in the prepartum period only. No other differences were observed between muscle group or treatment in either the pre- or postpartum period. Because there were no major differences between the cows in response to an IVGTT, we cannot conclude that glucose metabolism is a mechanism to explain differences in production responses observed. IVGTT cannot measure peripheral tissue insulin sensitivity, which is a limitation of this assessment, so our conclusions cannot assess if muscle reserves or BCVFA treatment impact peripheral tissue insulin sensitivity response. These results highlight that the amount of muscle plays a key role in the production responses observed in early lactation and that providing a BCVFA supplement could increase DMI during a period of negative nutrient balance and improve rumen efficiency.</p>

Certaines stratégies alimentaires sont actuellement considérées pour remplacer l’usage des antimicrobiens dans les fermes porcines. Les objectifs de cette étude étaient d'évaluer l'effet de la granulométrie et de la texture des aliments sur les concentrations d'acides gras volatils intestinaux, la composition des populations pathogènes et commensales d’E. coli et sur les performances de croissance des porcs. Des porcs d'engraissement (n= 840) ont reçu l'une des six diètes suivantes: moulée texturée 500, 750 et 1250 µm et moulée cubée 500, 750 et 1250 µm. Le gain de poids a été mesuré à chaque changement de formulation de moulée. À l'abattoir, les contenus du caecum et du côlon de 165 porcs ont été échantillonnés pour le dénombrement des E. coli par PCR quantitatif (qPCR) et pour la quantification des AGV. Le gène yccT a été utilisé pour dénombrer les E. coli totaux. Une diminution du taux de conversion alimentaire a été associée avec la moulée cubée et/ou la moulée de 500 µm. Les concentrations d’acide propionique et butyrique, et ce tant au niveau du caecum que du côlon, étaient plus élevées chez les porcs recevant de la moulée texturée que chez ceux recevant de la moulée cubée. Du point de vue de la granulométrie, les concentrations caecales et du côlon d’acide butyrique étaient plus élevées chez les porcs alimentés avec de la moulée de 1250 µm que chez ceux recevant de la moulée de 500 µm. D'autre part, les niveaux intestinaux d’E. coli totaux étaient plus élevés pour les porcs nourris avec de la moulée cubée que pour ceux ayant reçu de la moulée texturée. Les résultats ont montré que la moulée texturée est associée à des performances de croissance plus faibles mais à des changements intestinaux favorables. / Natural approaches, such as dietary modifications, are now being considered to replace antimicrobials for the control of bacterial infections in pigs. The objectives of this study were to assess the effect of feed size and texture on intestinal Volatile fatty acids profiles and concentrations, E. coli populations, and on growth performance. Fattening pigs (n=840) received one of six different diets: mash feed 500, 750 and 1250 µm and pellet feed 500, 750 and 1250 µm. Weight gain of pigs was monitored for each diet formulation over the fattening period. At the slaughterhouse, caecal and colon contents from 165 pigs were sampled for enumeration of E. coli by quantitative PCR (qPCR) and for acetic, propionic and butyric acid quantification. The yccT gene was used to enumerate total E. coli. A decrease in feed conversion associated with pellet texture and/or 500 µm particle size was observed for each diet formulation. In addition, caecal and colon propionic acid concentrations and caecal and colon butyric acid concentrations were lower for pigs receiving pellet rather than mash feed. Moreover, caecal (p=0,0208) and colon (p=0,0006) butyric acid concentrations were higher for pigs receiving a feed with a 1250 µm rather than 500 µm particle size. For total E. coli enumeration, caecal (p=0,01) and colon (p=0,04) yccT gene copies were higher for pigs receiving pellet rather than for those receiving mash feed. Taken together, results showed that mash feed is associated with favourable intestinal changes (VFA levels) and with a reduction of E. coli in the pig.

A análise de alimentos para ruminantes consite na caracterização da sua composição química, que varia em função da edafoclimatologia e manejo agrotécnico ao longo do tempo. O manejo de desfolhação (28 e 42 dias), o genótipo e estações constituem fonte de variação da degradabilidade in vitro da matéria seca (DIVMS), acúmulo de gases no rúmen (PG), metano (CH4) e do perfil dos ácidos graxos de cadeia curta (AGCC), com reflexos na eficiência da produção animal. A introdução dessas práticas no cultivo dos gêneros Brachiarias e Cynodon pode disponibilizar forragem de boa qualidade nutricional, com baixos teores de fibra. As dietas baseadas nas gramíneas tropicais são ricas e fibra e trazem controvérsias nos sistemas de produção de ruminantes, pela emissão de dióxido de carbono (CO2) e de CH4 entéricos, com efeito de estufa (GEE) na atmosfera, proposto para reduzir pela comunidade internacional. Contudo, a mitigação dos GEE exige a quantificação da sua produção por área, animal e por ano. Econmicamente, o CH4 enterico é uma perda de 2-12% de energia que poderia ser convertida em produto animal. O objetivo do estudo foi descrever e explicar os efeitos da frequência de desfolhação (28 e 42 dias), genótipo e estações do ano na composição química, DIVMS, PG, CH4, no perfil dos AGCC e no fator de partição (FP) da Brachiaria brizantha cv. Marandu, Brachiaria híbrida cv. Mulato II (Convert HD 364&reg;) e do Tifton 85 (Cynodon spp.), cultivados em 2012/2013. A matéria seca (MS%) da forragem cortada a cada 42 dias aumentou nos dois anos de experimento e diminuiu para o Marandu e o Mulato II. A matéria mineral (MM) na forragem aumentou em corte de 42 dias e no 2º ano. A matéria orgânica (MO) foi maior para o Marandu e Mulato II nas duas frequências de corte, comparada com a do Tifton 85. As médias mais baixas de fibra em detergente neutro (FDN), ácido (FDA) e extrato etéreo (EE) foram observadas no corte de 28 dias segundo a ordem: FDN, MM, FDA Tifton 85 >Marandu > Mulato e para DIVMS foi Marandu > Mulato II > Tifton 85 no outono-inverno. A PG foi superior para todos os genótipos na primavera-verão em relação ao outono-inverno. Contrariamente, o CH4 teve aumentos de 11,7% (Marandu), 29,2% (Mulato) e 41,4% (Tifton 85), no outono-inverno. O volume de CH4 das espécies foi diferente nas colheitas de 28 d de rebrotação. O FP variou pouco entre frequências, mas sim com a as estações do ano. A relação estequiométrica do acetato, propionato, e butirato foi 6,2:3,0:1,0 (estação) a 7,5:3,2:1,0 (ano). As relações acetato: propionato e TAGCC: MSD tiveram padrão de variação constante ao longo dos tratamentos. Conclui-se que a DIVMS e o perfil de AGCC são menos afetados pelas estações do ano do que pelas frequências de corte e estas praticas tendem a melhorar a qualidade da forragem colhida na frequência de rebrotação mais longa. Os teores de PB, CC, MO, EE foram altos nas duas frequências de corte e no inverno. / Ruminant feed analysis characterizes nutritional quality as described by its chemical composition (CC). The CC of forage grasses, however, varies with edaphoclimate, agronomic management (soil fertility, irrigation and harvesting frequency), which can change dry matter degradability (DMD), rumen gas production (GP), CH4, and volatile fatty acids (VTA) profile in the rumen, impacting animal production efficiency. The introduction of such practices in Brachiaria and Cynodon spp. Production system may enhance the production of green, high quality and nutritional forage. Hydrolyzed forage results in VFA, the energy that generate milk and meat. However, tropical grasses diets, rich in fiber, contribute for green house gas (GHG) and bring controversies in the system associated to the atmosphere global warming.The international community calls for GHG reduction quantifying the amount produced in the system. Economically, the emission of CH4 by the animal represents 2-12% energy loss that could be converted into animal products. The objective of the study is to describe and explain the effects of clipping frequency (28 and 42 days interval), season and year on CC, GP, CH4, in vitro DMD and PF of Brachiaria brizantha cv. Marandu, Brachiaria Hybrid cv. Mulato II (Convert HD 364&reg;) and Tifton 85 [Cynodon dactylon (L.)], under intensive management, form 2011 to 2013. Forage desfoliated at 42 days (d) intervals increased dry matter (DM) and ash (MM) content of Tifton 85 and Brachiaria sp., respectively. Organic matter (MO) of Marandu and Mulato II is high on both cutting frequencies compared to that of Tifton 85.The lowest NDF, ADF and EE were recorded at 28d forage regrowth. The ranking of NDF, MM, FDA for forage was Tifton 85> Marandu> Mulato) and IVDMD (Marandu> Mulato II> Tifton 85) in autumn-winter. Gas production was high for all species in summer-spring than autumn-winter, while CH4 increased 11.7% (Marandu), 29.2% (Mulato II) and 41.4% (Tifton 85), in autumn-winter. Harvesting at 28 d interval changed CH4 yield for the species. The FP varied sitghtly between frequencies, but with the seasons. The averages of acetic, propionic, butyric acids followed stachiometric relationship of 6.2: 3.0: 1.0 to 7.5: 3.2: 1.0 for season and year. The acetate to propionate and AGCC to MSD ratio had constant pattern but changed over treatments. We conclude that fertilized and irrigated grasses\'s IVDMD and VFA are less affected by the seasons than the harvesting frequencies and these practices improve the quality of harvested forage in both regrowth frequencies. Forage PB, CC, MO and EE are higher in both summer and winter as well as in the long cutting interval.

La production de microalgues en hétérotrophie présente plusieurs avantages pour la production de biocarburants par rapport à la production autotrophe, comme une productivité plus importante en termes de biomasse et de lipides. Cependant, le développement industriel de ce procédé est limité par les coûts de productions associés au substrat organique (i.e. glucose) et à ceux liés à la stérilisation des fermenteurs. Les effluents de fermentation sombre, composés principalement d’acétate et de butyrate, pourraient être utilisés comme milieux de culture peu onéreux pour la culture hétérotrophe ou mixotrophe de microalgues. Les objectifs de cette thèse étaient i) de mieux appréhender la croissance algale sur des mélanges variés d’acétate et de butyrate en fonction de la présence ou l’absence de lumière et de la température de croissance et ii) d’évaluer la faisabilité d’utiliser des effluents de fermentation non stérilisés pour soutenir la croissance de microalgues oléagineuses. Tout d’abord, un modèle basé sur des bilans de masse a été construit afin de caractériser la croissance hétérotrophe de Chlorella sorokiniana et Auxenochlorella protothecoides (taux de croissance et rendements) sur des mélanges d’acétate et de butyrate. Les résultats ont montré que le rapport acétate:butyrate et la concentration en butyrate étaient deux paramètres clés pour soutenir la croissance hétérotrophe. Puis, il a été démontré que la présence de lumière et l’utilisation d’une température suboptimale (30 °C) pour la croissance algale permettaient de réduire l’inhibition du butyrate en permettant une production de biomasse autotrophe ou en améliorant la croissance sur acétate. Enfin, il a été montré que les microalgues peuvent être compétitives sur l’acétate lors de la croissance sur des effluents bruts de fermentation sombre en présence de bactéries fermentaires, grâce à la croissance rapide des microalgues sur acétate (1.75 j-1) et à un changement drastique des conditions de culture peu favorables à la croissance des bactéries d’origine fermentaire. / Growing microalgae in heterotrophic mode present several advantages over autotrophic mode such as a higher productivity in terms of biomass and lipids for biofuels production. Nevertheless, this process is limited by the production cost associated with the organic substrate (i.e. glucose) and fermenters sterilization costs. Dark fermentation effluents, mainly composed of acetate and butyrate, could be used as a low-cost medium to grow microalgae heterotrophically or mixotrophically. The aims of this PhD were i) to optimize microalgae growth on various mixtures of fermentations metabolites using the presence or absence light and different cultivation temperatures and ii) to assess the feasibility of using unsterilized fermentation effluents. First, a model based on mass balance was built to characterize heterotrophic growth rates and yields when Chlorella sorokiniana and Auxenochlorella protothecoides were supplemented with different mixtures of acetate and butyrate. Results showed that the acetate:butyrate ratio and the butyrate concentration per se were two key parameters for promoting heterotrophic growth. Then, further studies showed that the presence of light and the use of suboptimal temperature (30 °C) could reduce the butyrate inhibition on growth by either triggering autotrophic production of biomass or enhancing growth on acetate. Finally, it was shown that microalgae could outcompete fermentation bacteria for acetate when growing on raw dark fermentation effluents, thanks to a fast algal growth on acetate (1.75 d-1) and a drastic change of culture conditions to the detrimental of bacterial growth.

A matéria orgânica presente nas águas residuárias representa uma matéria prima barata para integrar o processo de digestão anaeróbia e gerar fontes de energia alternativas, como o hidrogênio. O hidrogênio pode substituir os combustíveis fósseis não renováveis e diminuir as emissões de gases do efeito estufa responsável por grande parte da mudança climática global. A combustão do hidrogênio com o oxigênio produz somente vapor d\'água e representa 2,75 vezes mais conteúdo de energia que qualquer hidrocarboneto. Além disso, os ácidos orgânicos gerados no processo podem ser utilizados como fonte de carbono para a produção de metano, hidrogênio por organismos fototróficos e biopolímeros e, para a remoção de nutrientes biológicos. A produção de hidrogênio tem sido estudada em reatores de crescimento suspenso e de crescimento imobilizado como o reator anaeróbio de manta de lodo (UASB), reator anaeróbio de leito fixo e reator anaeróbio de leito fluidificado. O objetivo desse estudo foi avaliar a produção de hidrogênio e ácidos orgânicos em um reator anaeróbio de leito fluidificado contendo argila expandida como material suporte. O reator tinha 190 cm de altura, 5 cm de diâmetro interno e um volume de 4192 \'CM POT.3\'. O reator foi inoculado com lodo pré-tratado termicamente a 90°C por 10 minutos para o enriquecimento do inóculo acidogênico. O sistema foi operado com tempo de detenção hidráulica (TDH) de 8, 6, 4, 2 e 1 h e foi alimentado com água residuária sintética contendo 2000 mg/L de glicose. A operação contínua teve duração de 178 dias, e em cada TDH o reator foi operado durante, aproximadamente, 30 dias. A 30°C e pH de 3,8, cerca de 92 a 98% da glicose foi removida. A diminuição do TDH de 8 para 1 h conduziu a um aumento na velocidade média de produção de hidrogênio volumétrico de 0,28 para 1,15 L/h.L, respectivamente. A máxima velocidade de produção de hidrogênio, de 1,15 L/h.L, foi obtida em TDH de 1 h. Em geral, o rendimento de produção de hidrogênio aumentou com a diminuição do TDH, de 1,84 mols \'H IND.2\'/mol glicose em TDH de 8 h para 2,29 mols \'H IND.2\'/mol glicose em TDH de 2 h. O biogás não continha metano e o conteúdo máximo de hidrogênio foi de 37% em TDH de 2 h. Foi observada uma correlação linear entre a velocidade de produção de hidrogênio e a taxa de carregamento orgânico (TCO). Ainda, a TCO máxima de 104 kg/\'M POT.3\'.d não inibiu a produção de hidrogênio. Os maiores metabólitos solúveis foram o ácido butírico (44,64-52,52%) e o ácido acético (41,17-47,48%) com insignificante concentração de etanol (menor do que 10%). A produção de hidrogênio estável e a porcentagem de ácido butírico sobre o ácido acético indicam que o tratamento térmico do lodo foi efetivo e uma típica fermentação do tipo ácido butírico pode ser assegurada por Clostridium. Além disso, foram alcançados bons desempenhos de produção de hidrogênio sob condições de pH em torno de 4,0. / Organic materials in the wastewater can be an inexpensive raw material for integrating fermentation process and suppling renewable energy source such as hydrogen. Nevertheless hydrogen can replace fossil fuels and reduce greenhouse gases responsible for global climate change. The hydrogen combustion with oxygen produces only water vapor and hydrogen gas has 2.75 times the energy content of any hydrocarbons. Furthermore, the volatile fatty acids production can be used as carbon source for methane, photosynthetic hydrogen, bioplastics production and biological nutrients removal. The hydrogen production was carried out in suspend cell system, anaerobic sludge blanket reactor, fixed bed reactors and fluidized bed reactors. The objective of this study was to evaluate the hydrogen and volatile fatty acids production in an anaerobic fluidized bed reactor containing expanded clay like a material to immobilize acclimated anaerobic sludge. The reactor has presented 190 cm high, 5 cm in internal diameter and volume of 4192 \'CM POT.3\'. The reactor was seeded with a heat treated sludge at 90°C for 10 minutes for enriching \'H IND.2\' producing inoculum and supressing methanogens. The system was operated at hydraulic retention time (HRT) of 8, 6, 4, 2 and 1 h and was fed with syntetic wastewater containing 2.000 mg glucose/L. The continuous operation lasted 178 days, and in each TDH the reactor was operated for approximately 30 days. At 30°C and pH 3.8, about 92-98% of glucose was removed. The decrease of HRT from 8.0 to 1.0 led to a marked increase in the mean volumetric hydrogen production rate from 0.28 to 1.15 L/h.L, respectively. The maximum volumetric hydrogen production rate, 1.15 L/h.L, was obtained at HRT of 1.0 h. In general, the hydrogen yield increase from 1.84 mol \'H IND.2\'/mol glucose at HRT of 8.0 h to 2.29 mol \'H IND.2\'/mol glucose at HRT of 2.0 h. The biogas was free methane and the hydrogen content accounted for 37% at HRT of 2.0 h. There was a linear correlation between the hydrogen production rate and the organic loading rate (OLR) and the maximum OLR, 104 Kg/\'M POT.3\'.d, is not expected to inhibit hydrogen production. The major soluble products were butyric acid (44.64- 52.52%) and acetic acid (41.17-47.48%) further ethanol was relatively insignificant (less than 10%). The stable hydrogen production and the percentagen of butyric acid over acetic acid indicated the heat treated sludge was effective and a typical butyrate type fermentation was achieved by Clostridium. In addition, hydrogen production could be performed under pH conditions near 4.0.

Polyhydroxyalkanoates (PHA) are a family of biodegradable polyesters naturally synthesised by some bacteria and archaea. PHA have high industrial value as bioplastics for packaging and biomedical applications. However, their broader use is hindered by high production costs and uncontrolled variation of polymer properties. The extreme halophile Haloferax mediterranei shows bioprocess advantages that can be exploited for the low cost production of the PHA copolymer Poly(3-hydroxbutyrate-co-3-hydroxyvalterate) (PHBV). The focus of this thesis is to identify process variables responsible for the uncontrolled variation of PHA properties in order to progress towards the robust production of PHBV using H. mediterranei. The outcome of the investigation is a novel cultivation strategy for the reliable synthesis of PHBV copolymers with controlled composition and microstructure showing minor differences in material characteristics. Initially, growth kinetics and PHBV synthesis were characterised under nitrogen-excess and nitrogen-limiting conditions in ammonium and for the first time, nitrate. The nitrogen source and concentration influenced PHBV accumulation and variations in polymer composition were observed with ammonium, highlighting the importance of the control of cultivation conditions. Volatile fatty acids (VFA) were found to be a more direct approach to determine PHBV composition and for the first time were used as substrates in H. mediterranei cultures. When the cells were grown in C4:0/C5:0 mixtures, the 3HV fraction in the PHBV was proportional to the percentage of C5:0 in the feed mixture, allowing the synthesis of copolymers with a predefined composition ranging from pure PHB to pure PHV. The cultivation strategy proved effective for the synthesis of HV rich PHBV, which is not easily obtained due to the 3HV precursor toxicity. The polymer microstructure was controlled using different feeding strategies: co-feeding generated random copolymers, while sequential feeding created block and blend copolymers. The synthesis of block copolymers is of interest because the materials show enhanced yield strength and mechanical strength, making such materials more suitable for commodity uses. Bespoke random, block, and blend copolymers with 0−100 mol% 3HV were synthesized and their thermal and mechanical properties studied. Lastly, high temperature cultivation and several surfactants were tested to enhance the production of bespoke PHBV from VFA. PHBV productivity and accumulation was greatly improved in a fed-batch bioreactor fermentation at 37°C with Tween-80 and the maximum PHBV content 58.9% was obtained. The polymers from shake-flasks and from bioreactors showed minor variations in their material properties, demonstrating the scalability and the robustness of the process developed. Further understanding of the different process variables affecting polymer synthesis and composition was gained in this thesis. It is now possible to produce PHBV with controllable composition, microstructure and minor differences in material characteristics. The novel and robust production strategy developed address the bioprocess challenge of minimising the uncontrolled variation of polymer characteristics that is currently hindering the wider use of PHA hence allowing the production of high quality polymers for commodity goods, packaging and biomedical applications.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-08T13:12:12Z No. of bitstreams: 1 Elizabeth Hortencio Ferreira Lima.pdf: 1462177 bytes, checksum: 77d2db37110891a6e2a4fd1fdafe1015 (MD5) / Made available in DSpace on 2017-02-08T13:12:13Z (GMT). No. of bitstreams: 1 Elizabeth Hortencio Ferreira Lima.pdf: 1462177 bytes, checksum: 77d2db37110891a6e2a4fd1fdafe1015 (MD5) Previous issue date: 2013-02-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / This study aimed to investigate the effect of administration of sodium monensin in the diet on blood metabolites and rumen of ewes before, during and pospartum, due to the complexity that this period is due to anatomical changes, physiological and metabolic changes in female to meet the higher demand for energy growth of the fetus and lactation. A prospective study was conducted involving 13 sheep, in order to evaluate the effect of monensin supplemented from 60 days before calving (dap), on the hormonal and metabolic profile in periods of antepartum and postpartum. We used 13 Santa Inês sheep, pregnant, clinically healthy. Two groups were formed randomly, a control group receiving bulky, balanced feed and mineral salt and another group that received beyond bulky, balanced ration containing 30 mg monensin / day and mineral salt with monensin. Blood and ruminal fluid were collected at 60, 50, 40, 30, 20, 10 days before delivery, at birth and at 10, 20 and 30 days postpartum. We conducted the CBC and biochemical measurement of the following variables: glucose, non-esterified fatty acids (NEFA), β-hydroxybutyrate, cholesterol, triglycerides, fructosamine, total protein, albumin, urea, creatinine and research ketone bodies in urine. Enzyme activity by aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), creatine kinase (CK) and hormonal determinations of cortisol and insulin. In the analysis of ruminal fluid pH were determined, proof of reduction of methylene blue (PRAM), the infusoria, the chloride content and the concentration of volatile fatty acids (VFA) (acetic, propionic and butyric). Statistical analysis was employed ANOVA and correlation study (P<0,05). With regard to haematological values in different physiological periods studied, there was no significant difference (P> 0,05), except for the white blood count revealed a mild leukocytosis with neutrophilia at birth significantly (P <0,05) in two groups. The enzymatic activity of AST levels were more elevated in the control group (P<0,05) during the postpartum period and significant differences were also recorded between groups, no 10 dpp and 20 dpp (P<0,05) and higher in the control group compared to monensin. There was no difference in cholesterol (P>0,05). Effect of time (P<0,05) was observed with respect to triglycerides in both groups, being higher in the pre-partum. Monensin significantly (P<0,05) increased concentration of propionate in the rumen, fructosamine and insulin. Little effect was observed for glucose, NEFA and BHB. No treatment effect (P>0,05) was observed for pH, chloride and diatomaceous the rumen, and the protein profile. The administration of monensin in ewe in the period before the birth did not influence the indicators hematologic and enzymatic activity, but caused improvement in some indicators of energy balance, this result is considered important for the prevention of metabolic diseases associated with energy deficit that occurs this period, such as toxemia of pregnancy. / Este estudo teve por objetivo estudar o efeito da administração da monensina sódica na dieta sobre os metabólitos sanguíneos e ruminais de ovelhas antes, durante e após o parto, em razão da complexidade que representa este período devido às modificações anatômicas, fisiológicas e metabólicas na fêmea para atender a maior demanda energética para o crescimento dos fetos e a lactação. Um estudo prospectivo foi realizado envolvendo 13 ovelhas, com o intuito de avaliar o efeito da monensina, suplementada a partir de 60 dias antes do parto (dap), sobre o perfil metabólico e hormonal nos períodos do pré-parto e pós-parto. Foram utilizadas 13 ovelhas da raça Santa Inês, prenhes, clinicamente sadias. Dois grupos foram formados aleatoriamente, um grupo controle recebendo volumoso, ração balanceada e sal mineral e outro grupo que recebeu além do volumoso, ração balanceada contendo 30 mg de monensina/dia e sal mineral com monensina. Amostras de sangue e fluido ruminal foram colhidas aos 60, 50, 40, 30, 20, 10 dias antes do parto, no momento do parto e aos 10, 20 e 30 dias pós-parto. Foi realizado o hemograma e mensuração das seguintes variáveis bioquímicas: glicose, ácidos graxos não esterificados (AGNE), β-hidroxibutirato, colesterol, triglicerídeos, frutosamina, proteína total, albumina, uréia, creatinina e pesquisa de corpos cetônicos na urina. Atividade enzimática através da aspartato aminotransferase (AST), fosfatase alcalina (FA), gama glutamiltransferase (GGT), creatino quinase (CK) e determinações hormonais de cortisol e a insulina. Na análise do fluido ruminal foram determinados o pH, a prova de redução do azul de metileno (PRAM), os infusórios, o teor de cloretos e a concentração dos ácidos graxos voláteis (AGV) (acético, propiônico e butírico). A análise estatística foi empregada a ANOVA e um estudo de correlação (p<0,05). Com relação aos valores hematológicos nos diferentes períodos fisiológicos estudados, não foi observado diferença significativa (P>0,05), com exceção do leucograma que revelou uma discreta leucocitose por neutrofilia no momento do parto de forma significativa (P<0,05) nos dois grupos. A atividade enzimática da AST apresentou-se mais elevada no grupo controle (P<0,05) durante o pós-parto e diferenças significativas também foram registradas entre os grupos, no10 dpp e 20 dpp (P<0,05) sendo mais elevada no grupo controle quando comparada ao monensina. Não houve diferença nos valores de colesterol (P>0,05). Efeito de momento (P<0,05) foi observado com relação aos triglicerídeos nos dois grupos, sendo mais elevados no período pré-parto. A monensina influenciou significativamente (P<0,05) elevando a concentração do propionato no rúmen, frutosamina e insulina. Pouco efeito foi constatado para a glicose, BHB e AGNE. Nenhum efeito do tratamento (P>0,05) foi observado para o pH, teor de cloretos e os infusórios do rúmen, e para o perfil proteico. A administração da monensina nas ovelhas no período que antecede ao parto não influenciou os indicadores hematológicos e a atividade enzimática, porém provocou melhora em alguns indicadores do balanço energético, este resultado é considerado importante para a prevenção de doenças metabólicas, associadas ao déficit energético que ocorre neste período, como a toxemia da prenhez.

This thesis is focused on the development of an analytical method for the determination of volatile fatty acids in aqueous samples using GC-MS/MS. The theoretical section provides general information on a selected group of acids, their significance in the biogas plants, the means of their derivatization and the analytical techniques used. The practical part mainly deals with optimizition of various parameters in the derivatization reaction to achieve optimal conditions. Finally, the analytical method was successfully validated and applied to the analysis of a real sample.

Submitted by Silvana Teresinha Dornelles Studzinski (sstudzinski) on 2016-07-28T12:07:38Z No. of bitstreams: 1 Jamile Machado Hallam_.pdf: 1161483 bytes, checksum: 328ee4f618bd889a5e3570f303cdb9de (MD5) / Made available in DSpace on 2016-07-28T12:07:38Z (GMT). No. of bitstreams: 1 Jamile Machado Hallam_.pdf: 1161483 bytes, checksum: 328ee4f618bd889a5e3570f303cdb9de (MD5) Previous issue date: 2016-02-26 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / PROSUP - Programa de Suporte à Pós-Gradução de Instituições de Ensino Particulares / O aproveitamento energético obtido pela digestão anaeróbia é uma alternativa viável para o tratamento de resíduos alimentares, no entanto requer um estrito controle operacional. A escolha da melhor condição de partida pelo cálculo da relação substrato/inóculo (S/I) em termos de Sólidos voláteis (SV) pode facilitar estabilização do processo, resultando em elevada produtividade de biogás e teor de metano. Este estudo testou 5 diferentes relações de S/I em regime de batelada a 35oC, visando a estabilidade do processo e otimização da produção de biogás com alto teor de metano. Foi utilizado como substrato o resíduo coletado no restaurante universitário da Unisinos, e o como inóculo o lodo anaeróbio do reator UASB tratando esgoto doméstico da estação de tratamento de esgotos da mesma universidade. Os parâmetros monitorados foram pH, volume de biogás, teor de metano, alcalinidade e ácidos graxos voláteis. Todas as bateladas mantiveram-se dentro da faixa de pH recomendada pela literatura, entre 6,5 e 7,5, com exceção à batelada 5, que utilizou uma relação 1:1 S/I e onde o pH decaiu para 5,0. A produção de biogás em todas as condições estudadas teve sua máxima produção até o terceiro dia de inoculação e foi influenciada pela relação S/I aplicada, pois o aumento da quantidade de SV do substrato aumentou o rendimento e o teor de metano, com exceção da condição 1:1, onde houve a acidificação do sistema. O rendimento de biogás das condições 1:5, 1:4, 1:3, 1:2, 1:1 foram respectivamente 0,69, 0,75, 0,67, 0,89 e 0,18 L/g SV do substrato utilizado, e os teores médios de metano em cada batelada foram 69, 67, 65, 74 58%. Concluiu-se então, que a melhor relação substrato/inóculo para partida do processo foi 1:2, para a qual obteve-se 0,89 L de biogás/g SV do substrato com um teor de metano de 74%, mantendo uma relação AGV/ALC abaixo de 0,5, demonstrando a estabilidade do processo. / The energy recovery acquired through the anaerobic digestion is a viable alternative for treating food waste, though it requires an operational strict control. The choice of the best start up condition athwart the substratum /inoculum (S/I) ratios in terms of volatile solids (VS) may facilitate the process stabilization, resulting in increased production of biogas and methane content. The present study tested 5 different ratios of S/I in batch system at 35ºC, aiming the process stability and the optimization in the raised methane level biogas production. As the substrate, the residue collected at the university restaurant of Unisinos was used, and as inoculum the UASB reactor anaerobic sludge from the sewage wastewater treatment plant at the same university. The monitored parameters were pH, biogas amount, methane content, alkalinity and volatile fatty acids. All of the batches kept within the pH levels recommended by literature, between 6.5 and 7.5, exception to batch 5, which employed a relation of 1:1 substrate/inoculum and in which the pH level declined to 5.0. The biogas production in all of the studied conditions had its utmost production until the third day of inoculation and it was influenced by the S/I ration applied, since the increase of the substratum VS amount raised the yield and the methane content, exception to the condition 1:1 where there was an acidification of the system, and it was considered that the capacity of the commutation of substratum in methane was extrapolated. The biogas yields of the conditions 1:5, 1:4, 1:3, 1:2, 1:1 were, respectively, 0.69, 0.75, 0.67, 0.89 and 0.18 L/g VS of the substratum employed, and the average methane content in each batch were 69, 67, 65, 74 58%. The study shows that the best relation S/I to the process start-up was 1:2, to which it was obtained 0.89 L of biogas/g SV of the substratum with a methane content of 74% keeping the relation AGV/ALC under 0.5, demonstrating stability in the process.

The alpaca is a small South American camelid (SAC) that is an important production animal in Peru, especially among the highly impoverished communities of the high Andes, and raised for its fiber and meat. Alpacas are highly reliant on the microbes within their digestive tracts to digest the plant material they consume; volatile fatty acids (VFAs) are released as a byproduct of this microbial fermentation and used as a major source of energy by the alpaca. To explore optimal parameters for alpaca microbiome analysis, performed 16S rRNA gene surveys on alpaca C1 and fecal samples that had been extracted using one of three different DNA extraction methods (PowerFecal® DNA Isolation Kit (MO BIO); ZR Fecal DNA MiniPrep™ (Zymo); and a non-commercial extraction method called salting out) and amplified using one of two different polymerase enzyme mixes (AccuPrime™ Pfx SuperMix and 5 PRIME HotMasterMix). We found that choice of polymerase enzyme had a profound effect on the recovered microbiome, with the majority of 5 PRIME-amplified fecal samples failing to amplify. Extraction method had an effect on the recovered microbiome of fecal samples (but not C1 samples), with samples extracted using the MO BIO kit and the salting out method recovering different communities. The Zymo extraction kit returned microbial communities comparable to each of the other extraction methods. These results suggested that the AccuPrime enzyme and either the MO BIO or Zymo kits were optimal for alpaca gut microbiome analysis. We also performed two 16S rRNA gene surveys, the first from alpacas fed either a grass hay (GH) or alfalfa hay (AH) diet, and the second a C1 survey of alpacas fed two-week periods of mixed grass hay plus one of four supplements. We discovered body site and diet effects on the microbiota of alpacas fed either the GH or AH diet, with samples grouping by general body site (C1, small intestine, and distal intestine) and diet. However, we found no significant effect on the C1 microbiome of alpacas administered grain supplements. To study how energy extraction related to the microbiome, we correlated OTUs from GH/AH-fed alpaca with C1 VFA abundances. We discovered no significant correlations, and a 16S survey of low body condition (LBC) and good body condition (GBC) alpacas showed no difference in C1 microbial communities. We concluded that the microbiota of the alpaca digestive tract follow trends seen in microbiome studies of ruminants, but found no evidence of a relationship between body condition, energy extraction, and the C1 microbiome in alpacas.