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Resumo: O ácido lático foi quantificado no processo produtivo de várias destilarias do estado de São Paulo, com ênfase na fermentação, onde esse produto representa perdas de rendimento e afeta o metabolismo da levedura. As análises foram realizadas por meio de aparelhos dotados de tecnologia de enzima imobilizada, específicos para o isômero L(+) lactato e os resultados correlacionados com outros parâmetros do processo, obtidos pelas análises usuais. Na cana colhida, o teor de ácido lático variou entre 10,0 a 297,0 mg/L, de acordo com estágio de deterioração da matéria-prima. Nas destilarias anexas à fábrica de açúcar, o ácido lático foi detectado desde o caldo extraído nas moendas, onde se observou valor mínimo de 10 mg/L, até o mel final, no qual verificou-se teor superior a 8.800 mg/L. Nas fermentações contínuas registraram-se as maiores produções de ácido lático, superando 3700 mg/L - equivalente a mais de 6% (m/v) do álcool produzido, enquanto que em batelada alimentada conseguiu-se manter teores ao redor de 300 mg/L por 16 ciclos fermentativos. Bactérias isoladas de diferentes destilarias, e inoculadas individualmente em fermentações em laboratório, produziram quantidades distintas de ácido lático, de acordo com a espécie. Essa metodologia também foi empregada na avaliação da eficácia dos antibacterianos, usados freqüentemente nas destilarias brasileiras. Em fermentações tratadas com produtos específicos, a diminuição da atividade bacteriana determinou a queda na produção de ácido lático em mais de 70% em um ciclo fermentativo. / Abstract: The lactic acid was quantified in the ethanol production process of several distilleries of São Paulo State, especially in fermentation, where such product represents low yield and affects the metabolism of the yeast. The analysis were performed by equipments based on immobilized enzyme technology, specific for the isomer L(+) lactate and the results were correlated with others process parameters, got by the current methods. In the harvested cane, the lactic acid content varied from 10,0 to 297,0 mg/L, according to the deterioration of the raw material. In distilleries inside sugar factories, the lactic acid was quantified since the juice from crushers, where the registered values range from minimum of 10 mg/L until the molasses, in which values up to 8.800 mg/L were found. In the continuos fermentation it was registred the highest amount of lactic acid, up to 3.700 mg/L, corresponding to more than 6% of ethanol produced, while in fed batch it kept around 300 mg/L for 16 fermentaion cicles. Isolated from different distilleries, inoculated in laboratory fermentations, produced distinghished amount of lactic acid, according to the species. The method proposed also was used in the evaluation of efficacy of bactericidal, commonly used in the Brazilian distilleries. In fermentation treated with specific products, the reduction of bacterial activity leads the drop of lactic acid production up to 70% in one fermentation cicle. / Orientador: Dejanira de Franceschi de Angelis / Coorientador: Jonas Contiero / Banca: Octavio Antonio Valsechi / Banca: Marcia Justino Rossini Mutton / Mestre

Water and energy share an important relationship since it takes water to produce energy, and likewise, energy to pump, treat, and distribute water. This thesis explores the energy-water nexus in regards to electricity and transportation fuel production, as well as water treatment. It investigates how the Energy Independence and Security Act of 2007 might affect this interrelationship in the future since increases in corn cultivation for biofuels production are likely to lead to higher nitrate concentrations in US water reservoirs, which could trigger the requirement for additional energy consumption for drinking water treatment. The analysis indicates that advanced drinking water treatment might require an additional 2360 million kWh annually to treat drinking water currently exceeding the Environmental Protection Agency’s maximum contaminant level (MCL) limit of 10 mg per liter of nitrate-nitrogen. This is a 2100% increase in energy consumption for advanced water treatment to meet this MCL in comparison with surface water treatment alone. Although results indicate that most large surface and groundwater drinking water resources are not likely to exceed safe drinking water standards due to the expansion of corn-starch based ethanol production, smaller water reservoirs in agricultural regions are susceptible to nitrate contamination in the future. Consequently, these sources might require energy-intensive drinking water treatment to reduce nitrate levels below 10 mg per liter of nitrate-nitrogen. Based on these results, I conclude that projected increases in nitrate contamination in water may impact the energy consumed in the water treatment sector, because of the convergence of several related trends: (1) increasing cornstarch-based ethanol production, (2) increasing nutrient loading in surface water and groundwater resources as a consequence of increased corn-based ethanol production, (3) additional drinking water sources that exceed the MCL for nitrate, and (4) potentially more stringent drinking water standards for nitrate. / text

Goldfish subjected to an intermittent short-term aerobic exercise training regime prior to acute hypoxic exposure demonstrated a shift in hypoxia response. Intermittent aerobic training enhanced the aerobic potential of goldfish in the red muscle by increasing maximal activity of citrate synthase by 72% and reduced pyruvate kinase activity by 21% in white muscle. Across red and white muscle tissue, aerobic training caused a decrease in glycogen storage by 19% and 32%, respectively. Liver glycogen stores remained unchanged by training during normoxia. Subsequent hypoxic exposure demonstrated a significant training effect with a77% glycogen depletion in the liver of trained fish compared to a 53% depletion in untrained fish. Hypoxia caused glycogen depletion, glucose mobilization, and ATP depletion in trained and untrained fish muscle tissue. Meanwhile, the liver of trained recovered ATP slower than untrained fish and both liver and plasma had greater lactate accumulation by 1 h hypoxic recovery in trained fish. Alcohol dehydrogenase maximal activity of trained fish responded to hypoxia with a 50% reduction and trained white muscle significantly reduced alcohol dehydrogenase activity during hypoxic recovery. Ethanol was produced with and without training preconditioning in response to hypoxia in red muscle; however, trained fish white muscle showed an ethanol accumulation trend following training and 12 h hypoxia that was significantly cleared during recovery. Ethanol accumulation in white muscle of trained fish may reveal greater perturbation caused by training and hypoxia and/or some developed mechanism for ethanol retention. In effect, this training regime created a very different metabolic profile in goldfish such that during environmental oxygen limitation, trained fish may experience an enhanced metabolic perturbation and greater glycogen depletion which may compromise hypoxic tolerance. / Thesis (Master, Biology) -- Queen's University, 2011-09-30 13:25:36.148

Food prices have increased rapidly in recent years, and so has ethanol consumption. Some studies have claimed that there is a connection between those two. Net exporters of food tend to benefit from higher prices, while regions that are net importers of food, tend to be adversely affected. The large amount of poor countries in the second group justifies an investigation of the causes of increasing food prices. This thesis aims to contribute to the discussion, analyzing, theoretically and empirically, the impact that the diversion of feedstock from food to ethanol production has on food prices. The interaction between food prices and ethanol is first examined in a two-good (food and ethanol), one input (land) theoretical model. The outcome of this model is that an increase in ethanol productivity will have a positive impact on food prices, which is confirmed in the empirical test. We also found that increases in area allocated to produce sugarcane based ethanol in Brazil had depressing effects on relative food prices. No significant conclusion could be found on the effect of the area allocated to produce corn based ethanol in the United States.

Made available in DSpace on 2014-06-11T19:27:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-09-13Bitstream added on 2014-06-13T18:31:31Z : No. of bitstreams: 1 ventura_r_me_rcla.pdf: 536456 bytes, checksum: fc2eda253c87b88ee77bbe59f11812a1 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O ácido lático foi quantificado no processo produtivo de várias destilarias do estado de São Paulo, com ênfase na fermentação, onde esse produto representa perdas de rendimento e afeta o metabolismo da levedura. As análises foram realizadas por meio de aparelhos dotados de tecnologia de enzima imobilizada, específicos para o isômero L(+) lactato e os resultados correlacionados com outros parâmetros do processo, obtidos pelas análises usuais. Na cana colhida, o teor de ácido lático variou entre 10,0 a 297,0 mg/L, de acordo com estágio de deterioração da matéria-prima. Nas destilarias anexas à fábrica de açúcar, o ácido lático foi detectado desde o caldo extraído nas moendas, onde se observou valor mínimo de 10 mg/L, até o mel final, no qual verificou-se teor superior a 8.800 mg/L. Nas fermentações contínuas registraram-se as maiores produções de ácido lático, superando 3700 mg/L – equivalente a mais de 6% (m/v) do álcool produzido, enquanto que em batelada alimentada conseguiu-se manter teores ao redor de 300 mg/L por 16 ciclos fermentativos. Bactérias isoladas de diferentes destilarias, e inoculadas individualmente em fermentações em laboratório, produziram quantidades distintas de ácido lático, de acordo com a espécie. Essa metodologia também foi empregada na avaliação da eficácia dos antibacterianos, usados freqüentemente nas destilarias brasileiras. Em fermentações tratadas com produtos específicos, a diminuição da atividade bacteriana determinou a queda na produção de ácido lático em mais de 70% em um ciclo fermentativo. / The lactic acid was quantified in the ethanol production process of several distilleries of São Paulo State, especially in fermentation, where such product represents low yield and affects the metabolism of the yeast. The analysis were performed by equipments based on immobilized enzyme technology, specific for the isomer L(+) lactate and the results were correlated with others process parameters, got by the current methods. In the harvested cane, the lactic acid content varied from 10,0 to 297,0 mg/L, according to the deterioration of the raw material. In distilleries inside sugar factories, the lactic acid was quantified since the juice from crushers, where the registered values range from minimum of 10 mg/L until the molasses, in which values up to 8.800 mg/L were found. In the continuos fermentation it was registred the highest amount of lactic acid, up to 3.700 mg/L, corresponding to more than 6% of ethanol produced, while in fed batch it kept around 300 mg/L for 16 fermentaion cicles. Isolated from different distilleries, inoculated in laboratory fermentations, produced distinghished amount of lactic acid, according to the species. The method proposed also was used in the evaluation of efficacy of bactericidal, commonly used in the Brazilian distilleries. In fermentation treated with specific products, the reduction of bacterial activity leads the drop of lactic acid production up to 70% in one fermentation cicle.

The overall purpose of this study is to estimate how the rise in grain prices (especially corn prices) induced by ethanol production impacts U.S. catfish industry. Using monthly data from January 1996 to December 2007, an ARDL model and bounds testing procedure were used. The existence of cointegration between the feed price and its regressors and between the farm price and its regressors was found. Results show that the short- and long-run feed price elasticity with respect to corn prices were 0.224 and 0.075, respectively. It was found that energy is more important to catfish feed production than to farm level catfish production, and is more important to processor level production than to farm level production. Results further showed that catfish farmers will lose net returns because the estimated farm price elasticity with respect to feed prices was smaller than the necessary change that would keep net returns the same. The overall purpose of this study is to estimate how the rise in grain prices (especially corn prices) induced by ethanol production impacts catfish feed prices and catfish prices at the farm level. Using monthly data from January 1996 to December 2007, an ARDL model and bounds testing procedure were used to test the existence of cointegration between the variables of interest. The existence of cointegration between the feed price and its regressors and between the farm prices and its regressors was found. Estimation results show that the short- and long-run feed price elasticity with respect to corn prices were 0.224 and 0.075, respectively. Feed and farm price elasticities with respect to energy prices were highlighted. The results show that energy is more important to catfish feed production than to catfish production at the farm level, and energy is more important to catfish production at the processor level than to production at the farm level.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: There is a global demand for technologies to reduce ethanol levels in wine without compromising wine quality. While several chemical and physical methods have been developed to reduce ethanol in finished wine, the target of an industrially applicable biological solution has thus far not been met. Most attempted biological strategies have focused on developing new strains of the main fermentative organism, the yeast Saccharomyces cerevisiae. Gene modification approaches have primarily focused on partially redirecting yeast carbon metabolism away from ethanol production towards glycerol production. These techniques have met with some moderate success, thus the focus of the current study was to re-direct carbon flux towards trehalose production by moderate over-expression of the TPS1 gene. This gene encodes trehalose-6-phosphate synthase, which converts glucose 6-phosphate and UDPglucose to α,α-trehalose 6-phosphate. Previous data have shown that the overproduction of trehalose restricts hexokinase activity reducing the amount of glucose that enters glycolysis. Nevertheless, preliminary TPS1 over-expression studies using multiple copy plasmids have shown some promise, but also indicated significant negative impact on the general fermentation behaviour of strains. In order to reduce such negative impacts of excessive trehalose production, a new strategy consisting in increasing the expression of TPS1 only during specific growth phases and by a relatively minor degree was investigated. Our study employed a lowcopy number episomal vector to drive moderate over-expression of the TPS1 gene in the widely used industrial strain VIN13 at different stages during fermentation. The fermentations were performed in synthetic must with sugar levels representative of those found in real grape must. This, as well as the use of an industrial yeast strain, makes it easier to relate our results to real winemaking conditions. A reduction in fermentation capacity was observed for all transformed strains and controls. Expression profiles suggest that the DUT1 promoter certainly results in increased TPS1 expression (up to 40%) during early exponential growth phase compared to the wild type strain (VIN13). TPS1 expression under the control of the GIP2 promoter region showed increased expression levels during early stationary phase (up to 60%). Chemical analysis of the yeast and the must at the end after fermentation showed an increase in trehalose production =in line with the expression data of TPS1. Importantly, glycerol production was also slightly increased, but without affecting acetic acid levels for the transformed strains. Although ethanol yield is not significantly lower in the DUT1-TPDS1 strain, s statistically significantly lower ethanol yield is observed for over-expression under the GIP2 promotor. Increasing trehalose production during stationary phase appears therefore to be a more promising approach at lowering ethanol yield and redirecting flux away from ethanol production. This controlled, growth phase specific over expression suggests a unique approach of lowering ethanol yield while not impacting on the redox balance. / AFRIKAANSE OPSOMMING: Wêreldwyd is daar ‘n aanvraag na tegnologie wat die etanol vlakke in wyn kan verminder sonder om wyngehalte te benadeel. Terwyl verskeie chemiese en fisiese metodes ontwikkel is om etanol in die finale wynproduk te verminder, is die soeke na 'n industrieel gebaseerde biologiese oplossing tot dusver nie gevind nie. Meeste biologiese strategieë fokus op die ontwikkeling van nuwe rasse van die primêre fermentatiewe organisme, naamlik Saccharomyces cerevisiae. Geen modifikasie benaderings het hoofsaaklik gefokus op die gedeeltelike kanalisering van koolstof metabolisme weg van etanol produksie na gliserol produksie. Hierdie benadering is net matiglik suksesvol, dus is ons huidige fokus om koolstof te kanaliseer na trehalose produksie deur gematigde oor-uitdrukking van die TPS1 geen. Hierdie geen kodeer vir trehalose-6-fosfaat sintase, wat glukose-6-fosfaat en UDP-glukose omskakel na α, α-trehalose-6-fosfaat. Vorige data het getoon dat die oorproduksie van trehalose hexokinase aktiwiteit beperk en die hoeveelheid glukose wat glikolise binne gaan. Voorlopige TPS1 ooruitdrukking studies met behulp van multi-kopie plasmiede toon matige sukses, maar het ook ‘n negatiewe impak op die algemene fermentasie kapasiteit van die gis. Ten einde so 'n negatiewe impak van oormatige trehalose produksie te oorkom, is 'n nuwe strategie gevolg wat bestaan uit die verhoogde uitdrukking van die TPS1 geen slegs gedurende spesifieke groei fases met baie lae vlakke van oor-uitdrukking. Ons studie gebruik 'n lae-kopie episomale vektor met matige oor-uitdrukking van die TPS1 geen in die industriële ras VIN13 op verskillende stadiums tydens fermentasie. Die fermentasie is uitgevoer in sintetiese mos met suiker vlakke verteenwoordigend van dié van werklike wyn mos. Hierdie, sowel as die gebruik van 'n industriële gisras, maak dit makliker om ons resultate te vergelyk met regte wyn fermentasie kondisies. Verlaagde fermentasie kapasiteit is waargeneem vir alle getransformeerde stamme en hul kontroles. Geen uitdrukkings profiele dui op verhoogde TPS1 uitdrukking (tot 40%) onder beheer van die DUT1 promotor gedurende die vroeë eksponensiële groeifase wanneer vergelyk word met die wilde tiepe (VIN13). TPS1 uitdrukking onder die beheer van die GIP2 promotor het verhoogde uitdrukking van tot 60% gedurende die vroeë stasionêre fase. Chemiese analise van die gis aan die einde van fermentasie dui op ‘n toename in trehalose produksie wat korreleer met die uitdrukking profiele van TPS1. Gliserol produksie is ook effens verhoog, maar sonder ‘n toename in asynsuur vlakke vir die getransformeerde rasse. Alhoewel etanol opbrengs nie aansienlik laer vir die DUT1-TPS1 ras is nie, is etanol opbrengs vir die oor-uitdrukking onder beheer van die GIP2 promotor wel laer. Toenemende trehalose produksie gedurende stasionêre fase blyk dus 'n meer belowende benadering op die verlaging van etanol opbrengs en her-kanaliseering weg van etanol produksie. Hierdie benadering met die fokus op groeifase spesifieke oor-uitdrukking dui op 'n unieke strategie vir die verlaging van etanol opbrengs sonder om die redoks balans te beinvloed. / The NRF, IWBT and Stellenbosch University for funding

Dentre as leveduras selvagens mais comumente encontradas na fermentação alcoólica, destaca-se o gênero Saccharomyces apresentando colônias rugosas com células dispostas em cachos (pseudohifas). Este biótipo de levedura tem sempre sido associado com problemas na indústria, ocasionando queda no rendimento fermentativo. Sendo assim, o presente trabalho teve por objetivo realizar a caracterização genética, morfo-fisiológica e da resistência ao estresse de linhagens de Saccharomyces cerevisiae que apresentam colônias rugosas e mucosas, buscando alternativas que possam contribuir para o manejo dessas leveduras selvagens (rugosas) na fermentação alcoólica. Foram realizados testes de caracterização para crescimento invasivo, atividade killer, temperatura, pH, concentração de etanol e açúcar, presença de actidione, floculação e capacidade fermentativa, utilizando-se 22 linhagens de leveduras (11 rugosas e 11 mucosas) selecionadas previamente por seqüenciamento da região ITS do DNA ribossomal. O efeito do tratamento ácido em diferentes valores de pH sobre o crescimento de duas linhagens (52 rugosa e PE-02 mucosa) foi também avaliado, seguido do monitoramento do crescimento em meio de caldo de cana após tratamento ácido. Em seguida, testes fermentativos em meio de caldo de cana foram conduzidos em culturas puras e mista dessas linhagens. Os testes de caracterização morfofisiológica mostraram que a invasividade em meio YEPD Agar ocorreu em baixa freqüência dentre as 22 leveduras testadas; dez dentre onze leveduras rugosas apresentaram taxa de floculação expressiva, e entre as mucosas a floculação foi praticamente inexistente; nenhuma das linhagens apresentou produção de toxina killer; as linhagens com colônias rugosas apresentaram capacidade fermentativa significativamente inferior às linhagens com colônias mucosas, em sistema de batelada com ciclo único de 48 horas em meio de caldo de cana, demonstrando velocidade mais lenta de fermentação. Quanto à resistência ao estresse por temperatura, pH, etanol e concentração de açúcar, as linhagens mucosas tiveram maior resistência à acidez do meio, enquanto as linhagens rugosas foram mais resistentes às concentrações elevadas de etanol e açúcar. Nenhuma levedura foi resistente ao actidione. A análise genética, através dos loci microssatélites, revelou a presença de dois grupos principais relacionados geneticamente, sendo o primeiro ramo constituído principalmente de colônias rugosas (67%), contendo, no entanto, a linhagem PE-02, apresentando linhagens com alta taxa de floculação e tolerância às altas concentrações de açúcar e etanol; o outro grupo (com três subgrupos) compreendeu principalmente colônias mucosas, apresentando pouca resistência às situações estressantes aqui estudadas. A levedura com colônia rugosa (linhagem 52) foi bastante afetada pelo tratamento ácido em valores de pH 1,0 e 1,5, ao contrário da levedura com colônia mucosa (PE-02). A fermentação em sistema de batelada com reciclo celular e tratamento ácido conduzido em pH 1,5 teve efeito sobre o crescimento da levedura rugosa quando em cultura mista com a levedura mucosa, não resultando em prejuízo à eficiência fermentativa, quando comparada com a cultura pura da PE-02. Em cultura pura da levedura rugosa, constatou-se eficiência fermentativa por volta de 60%, confirmando o baixo desempenho destas leveduras. O conhecimento das respostas das leveduras rugosas 12 às situações estressantes pode ajudar a manejar a fermentação alcoólica para minimizar o efeito da levedura contaminante. / Among the wild yeasts more commonly found in the alcoholic fermentation, wrinkled colonies with pseudohyphal morphology belonging to Saccharomyces genus are highlighted. This yeast biotype has been associated with industrial problems, resulting in the decrease of the fermentative yield. In this context, this work aimed to perform the genetic, morphological/physiological characterization and stress tolerance of Saccharomyces cerevisiae strains which exhibited wrinkled and mucous colonies, searching for alternatives that could contribute to the management of these wild yeasts (wrinkled colonies) in the alcoholic fermentation. Characterization tests for invasiveness in Agar medium, killer activity, temperature, pH, ethanol and sugar concentration, actidione, flocculation and fermentative capacity were carried out with 22 strains (11 wrinkled and 11 mucous colonies), which were screened previously by the sequencing of the ITS region of the ribosomal DNA. The effect of the acid treatment using different pH values over the growth of two strains (52 wrinked and PE-02 mucous) was also evaluated, following the growth monitoring in sugar cane juice after acid treatment. Fermentative tests in sugar cane juice were carried out with pure and mixed cultures of these strains. The morphological/physiological tests showed that the invasiveness in YEPD Agar medium occurred in low frequency among the 22 strains tested; ten out of eleven wrinked yeasts exhibited expressive flocculation, and among the mucous, the flocculation was near zero; none of the strains showed killer activity; the wrinkled colonies presented lower fermentative capacity comparing to mucous colonies, in a 48- hour cycle in batch system using sugar cane juice, with slower fermentation rate. Concerning the resistance to temperature, pH, ethanol and sugar concentration, the mucous colonies were more resistant to low pH, while the wrinkled colonies were more resistant to the elevated ethanol and sugar concentrations. None of the yeasts were resistant to actidione. The genetic analysis by microsatellite loci revealed the presence of two main genetic related groups, the first branch comprised mainly of wrinkled colonies (67%), containing however the PE-02 strain, showing strains with high flocculation rate and tolerance to high concentration of sugar and ethanol; the other group (with three subgroups) comprised mainly mucous colonies, showing lower resistance to the stressing conditions here studied. The yeast with wrinkled colony (strain 52) was severely affected by the acid treatment in pH values of 1.0 and 1.5, but the same did not occur with the mucous colony (PE-02). The batch fermentation with cell recycle and acid treatment in pH 1.5 had effect over the wrinkled yeast growth when in mixed culture with the mucous yeast, and did not impair the fermentative efficiency comparing to the pure culture of PE-02. In pure culture with the wrinkled colony, a fermentative efficiency as low as 60% was observed, confirming the low performance of these yeasts. The knowledge of the response to stressful conditions exhibited by the yeasts with wrinkled colonies can help to manage the alcoholic fermentation in order to minimize the effect of the contaminant yeast.