Doctor of Philosophy / Department of Grain Science and Industry / Yong Cheng Shi / This study focused on in-vitro digestibility and pasting properties of cross-linked (CL) starch with different amylose contents and different cross-linking levels, as well as physicochemical properties of extruded wheat flours with different amylose contents and functionality of amylopectin and cross-linking in improving the textural and physical properties of oat flour extrudates. Starch was CL by phosphorylation using a mixture of sodium trimetaphosphate and sodium tripolyphosphate at the ratio of 99:1 under alkaline condition. The digestibility of highly CL maize starches with different amylose contents was determined by Englyst, Available Carbohydrate Dietary Fiber and Association of Official Analytical Chemists (AOAC) Method 991.43 methods. CL waxy and normal maize starch granules swelled much more at higher temperatures, resulting in significantly lower total dietary fiber content than high-amylose maize starch. The alkali treatment study on low levels of cross-linking in starch affected the ratios of total bound phosphate esters which changed the degree of starch swelling, crytallinity and pasting properties of the CL starch. The study on the mechanism of the digestive enzymes and the extent of digestion during the incubation of CL wheat starch in AOAC Method 2009.01 showed progressive digestion after 16 h of incubation, which may not reflect in-vivo response in human. Extrusion of normal and waxy wheat flours resulted in the breakdown of starch and an increase in the insoluble protein, which affected the textural and structural properties of extrudate. High energy input played a major role in radial expansion of normal hard wheat extrudate, whereas higher amylopectin in soft waxy wheat flour was a dominant factor in determining the radial expansion when compared to normal soft wheat. Inclusion of soft waxy wheat in the oat flour formulations at 18% moisture content improved the textural and structural properties of extrudates. Low cross-linking level of CL waxy maize starch in oat flour formulation increased the void fraction and reduced the breaking strength of extrudates, whereas higher cross-linking levels of CL starches improved the resistant starch level on oat flour formulation but had very poor structural and textural properties.The study offers a good insight on the properties and digestibility of CL starch, as well as using low levels of CL starch to improve textural properties of nutritional extruded products. In addition, study on the extrusion of wheat flours with different amylose and protein contents provides knowledge on the influence of chemical compositions and energy input on the physico-chemical properties of extrudates.

Actinobactérias são microrganismos capazes de produzir compostos bioativos presentes em extratos brutos com ação antibiótica e ionófora. A hipótese deste estudo foi de que extratos brutos de actinobactérias apresentem eficácia similar à monensina sódica para modulação da fermentação ruminal; no aumento da digestibilidade da matéria seca e matéria orgânica, aumento da concentração de propionato e redução da produção de gases totais, da desaminação protéica e de metano. Portanto, o objetivo foi avaliar in vitro dois extratos brutos (AMC e Caat) sobre a fermentação ruminal (produção total de gases, de ácidos graxos de cadeia curta (AGCC) e de metano (CH4); digestibilidade in vitro da matéria seca (DIVMS), da matéria orgânica (DIVMO) e produção de nitrogênio amoniacal (N-NH3). O estudo foi dividido em duas fases; a primeira objetivou estabelecer a curva dose-resposta dos extratos e a segunda para avaliação com controle positivo de monensina sódica. Para ambas fases, foram utilizadas como doadoras de líquido ruminal (inóculo) três vacas da raça Holandesa em lactação, portadoras de fístula ruminal. As dietas fornecidas foram à base de silagem de milho, farelo de soja, ureia, milho moído e suplemento mineral, com relação volumoso:concentrado de 60:40. Amostras de fluido ruminal foram coletadas por meio de bomba de vácuo, armazenadas em garrafas e caixas térmicas e transportadas ao laboratório para incubação em frascos de vidro. Para a primeira fase, foram avaliadas 4 doses (0,3; 0,6; 0,9 e 1,20 mg) de 2 extratos (AMC e Caat) em ensaio in vitro de 72 horas, no qual foram mensuradas a produção de gás durante a incubação e, ao final, as concentrações de AGCC, DIVMS e DIVMO foram determinadas. Na segunda fase, o ensaio foi de 24 horas, e ao final foram feitas análises para CH4 e N-NH3, além de AGCC, DIVMS, DIVMO e produção de gás. O delineamento experimental adotado foi o inteiramente casualizado (DIC) para ambas as fases. Na primeira fase, a inclusão do Caat diminuiu linearmente a DIVMO após 72 hs e reduziu as concentrações de ácido isovalérico, butírico, da relação acetato:propionato e da produção de gás acumulado por 24, 48 e 72 h. Na segunda fase, a inclusão dos extratos causaram diminuição da produção de gás acumulada em 24 horas, sendo menor no tratamento monensina e no Caat. Houve efeito de inclusão do Caat e da monensina sobre a concentração de AGCC, aumentando a concentração de propionato e diminuindo a concentração de butirato, acetato e a relação acetato:propionato. A concentração de N-NH3 foi menor mediante inclusão do extrato Caat e da monensina enquanto que a concentração de CH4 foi maior para a inclusão dos extratos quando comparados com a inclusão de monensina, entretanto não foram maiores do que o controle. A inclusão do extrato Caat foi capaz de modificar a fermentação in vitro de forma semelhante à monensina sódica, mas não alterou a digestibilidade da fibra. Por outro lado, o AMC não alterou a fermentação ruminal, seja na digestibilidade da fibra, na produção de gases totais, metano, nitrogênio amoniacal e AGCC / Actinobacteria are microorganisms that can produce bioactive compounds present in crude extracts with antibiotic and ionophore activity. Our hypothesis was that actinobacteria crude extracts showed similar efficacy to monensin on ruminal fermentation; increasing the digestibility of dry matter and organic matter, increasing propionate concentration and reducing the total gas production, protein deamination and methane. Therefore, the objective was to evaluate two crude extracts (AMC and Caat) on in vitro ruminal fermentation (total gas, short chain fatty acids (SCFA) and methane (CH4) production; in vitro digestibility of dry matter (IVDDM), of organic matter (IVOMD) and ammonia nitrogen production (NH3-N). The study was divided into two phases: the first aimed to establish the dose-response curve of the extracts and the second aimed to evaluate a positive monensin control. For both, it was used three lactating Holstein cows with ruminal fistula as rumen fluid donors (inoculum). The diets were based on corn silage, soybean meal, urea, ground corn and mineral supplement under forage:concentrate ratio of 60:40. Ruminal fluid samples were collected by a vacuum pump, stored in bottles and coolers and transported to the laboratory for incubation in glass bottles. For the first phase, they were evaluated 4 doses (0.3; 0.6; 0.9 and 1.20 mg) of 2 extracts (AMC and Caat) on a 72 hours in vitro assay, which were measured gas production during incubation and in the end, SCFA, IVDMD and IVOMD concentrations were determined. In the second phase, it was a 24 hours assay, and include final analyzes for CH4 and NH3-N, plus SCFA, IVDMD, IVOMD and gas production. The experimental design was completely randomized (DIC) for both phases. In the first phase, the Caat inclusion linearly decreased IVOMD after 72 hours and reduced isovaleric acid and butyric acid concentration and acetate:propionate ratio and cumulative gas production by 24, 48 and 72 h. In the second stage, the inclusion of the extracts caused a reduction of the cumulative gas production in 24 hours, being lower in monensin and Caat treatments. There was an effect of Caat and monensin inclusion on SCFA concentration, increasing the propionate concentration and reducing the butyrate and acetate concentration and the acetate: propionate ratio. The NH3-N concentration was lower by Caat inclusion and monensin while the CH4 concentration was higher for the extracts inclusion as compared with monensin, however were not bigger than control. The Caat inclusion was able to modify the in vitro fermentation similarly to monensin, but did not affect the fiber digestibility. Moreover, AMC did not affect the rumen fermentation, either in fiber digestibility, total gas, methane, ammonia nitrogen and SCFA production

Made available in DSpace on 2014-06-11T19:23:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-11-09Bitstream added on 2014-06-13T20:11:03Z : No. of bitstreams: 1 cruz_ks_me_arafcf.pdf: 1084231 bytes, checksum: 51b3fe7285e63cfa32b590132f66f964 (MD5) / Universidade Estadual Paulista (UNESP) / O baru (Dipteryx alata Vog.), fruta nativa do Cerrado brasileiro, apresenta uma amêndoa com alto teor de proteínas, entretanto, informações sobre suas propriedades são ainda escassas. O objetivo do trabalho foi caracterizar algumas das propriedades físico-químicas da proteína dessa semente. Percentuais de 74,13; 81,56 e 76,50 de proteínas foram solubilizadas frente às diferentes proporções massa:volume 1:10, 1:30 e 1:50, respectivamente. Em relação aos tempos 30, 60 e 90 minutos de extração solubilizaram-se 78,59, 80,45 e 73,74% das proteínas totais, respectivamente. Frente às modificações de pH e concentração de NaCl, verificouse um comportamento típico da predominância de globulinas, caracterizado por máxima solubilidade em pHs 2,0 (87,9%) e acima de 8,0 (89,87%) e mínima solubilidade (8,5%) em pH 4,5, em água, e aumento junto ao ponto isoelétrico associado a crescente concentração salina no meio extrator. O fracionamento pelo critério de solubilidade em diferentes sistemas revelou teores de 13,97% de albuminas; 61,74% de globulinas, 0,48% de prolaminas, 3,25% de glutelinas e 4,61% de proteínas insolúveis. A digestibilidade in vitro, determinada pela sequência enzimática pepsina-pancreatina, mostrou 85,59% e 90,54% de hidrólise em relação à caseína para a farinha desengordurada e a globulina total da amêndoa, respectivamente. A proteólise da globulina majoritária obtida em Sephadex G-200, pelas enzimas tripisina, quimotripsina e pepsina, separadamente, mostrou percentuais de hidrólise, em relação à caseína, de 62,38, 17,92 e 71,38, respectivamente, após 120 minutos de incubação, sendo o progresso das proteólises acompanhados em SDS-PAGE. O perfil cromatográfico em colunas de filtração em gel mostrou a existência de uma forma protéica majoritária na fração globulina total que, em coluna de troca-iônica de DEAE-celulose, foi eluída... / The Baru (Dipteryx alata Vog.), a fruit native to the Brazilian Cerrado, has a seed with a high protein content, however, informations about its properties remain scarce. The aim of this study was to characterize some of the physicochemical properties of this seed protein. Protein percentage of 74.13, 81.56 and 76.50 were solubilized using the mass:volume ratios 1:10, 1:30 and 1:50, respectively. During the extraction times 30, 60 and 90 minutes were solubilized 78.59, 80.45 and 73.74% of total proteins, respectively. Changes in pH and salt concentration reveled a typical behavior of gobulin predominance, characterized by greater solubility at pH 2.0 (87.9%) and above 8.0 (89.87%) and lower solubility (8.5%) at pH 4.5 in water, and solubility improvement near the isoelectric pH by increasing salt concentration in the extractant. Fractionation according to protein solubility in different solvents showed 13.97, 61.74, 0.48, 3.25 and 4.61 percent of albumin, globulin, prolamin, glutelin and insoluble protein, respectively. The in vitro digestibility determined by pepsin-pancreatin enzimatic sequence showed 85.59% and 90.54% of hydrolysis, in relation to casein, for the defatted flour and total globulin from baru seed, respectively. After 120 minutes of incubation, proteolysis of the major globulin obtained from Sephadex G-200 by tripsin, chymotrypsin and pepsin, separately, showed hydrolysis porcentages, in relation to casein, of 62.38, 17.92 and 71.38, respectively, and the progress of these proteolysis was monitored by SDS-PAGE. The chromatographic profile in gel filtration columns reveled the existence of one major protein form in total globulin, which was eluted off the DEAE-celulose ionexchange column at 0.12 M NaCl concentration. The molecular weight (MW) of globulin was estimated at 405-436 kDa and its electrophoretic profile showed three main bands of MW 62-64, 66 and 73-76 kDa, and... (Complete abstract click electronic access below)

Orientador: Valdir Augusto Neves / Banca: José Paschoal Batistuti / Banca: Osvaldo de Freitas / Resumo: O baru (Dipteryx alata Vog.), fruta nativa do Cerrado brasileiro, apresenta uma amêndoa com alto teor de proteínas, entretanto, informações sobre suas propriedades são ainda escassas. O objetivo do trabalho foi caracterizar algumas das propriedades físico-químicas da proteína dessa semente. Percentuais de 74,13; 81,56 e 76,50 de proteínas foram solubilizadas frente às diferentes proporções massa:volume 1:10, 1:30 e 1:50, respectivamente. Em relação aos tempos 30, 60 e 90 minutos de extração solubilizaram-se 78,59, 80,45 e 73,74% das proteínas totais, respectivamente. Frente às modificações de pH e concentração de NaCl, verificouse um comportamento típico da predominância de globulinas, caracterizado por máxima solubilidade em pHs 2,0 (87,9%) e acima de 8,0 (89,87%) e mínima solubilidade (8,5%) em pH 4,5, em água, e aumento junto ao ponto isoelétrico associado a crescente concentração salina no meio extrator. O fracionamento pelo critério de solubilidade em diferentes sistemas revelou teores de 13,97% de albuminas; 61,74% de globulinas, 0,48% de prolaminas, 3,25% de glutelinas e 4,61% de proteínas insolúveis. A digestibilidade in vitro, determinada pela sequência enzimática pepsina-pancreatina, mostrou 85,59% e 90,54% de hidrólise em relação à caseína para a farinha desengordurada e a globulina total da amêndoa, respectivamente. A proteólise da globulina majoritária obtida em Sephadex G-200, pelas enzimas tripisina, quimotripsina e pepsina, separadamente, mostrou percentuais de hidrólise, em relação à caseína, de 62,38, 17,92 e 71,38, respectivamente, após 120 minutos de incubação, sendo o progresso das proteólises acompanhados em SDS-PAGE. O perfil cromatográfico em colunas de filtração em gel mostrou a existência de uma forma protéica majoritária na fração globulina total que, em coluna de troca-iônica de DEAE-celulose, foi eluída... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The Baru (Dipteryx alata Vog.), a fruit native to the Brazilian Cerrado, has a seed with a high protein content, however, informations about its properties remain scarce. The aim of this study was to characterize some of the physicochemical properties of this seed protein. Protein percentage of 74.13, 81.56 and 76.50 were solubilized using the mass:volume ratios 1:10, 1:30 and 1:50, respectively. During the extraction times 30, 60 and 90 minutes were solubilized 78.59, 80.45 and 73.74% of total proteins, respectively. Changes in pH and salt concentration reveled a typical behavior of gobulin predominance, characterized by greater solubility at pH 2.0 (87.9%) and above 8.0 (89.87%) and lower solubility (8.5%) at pH 4.5 in water, and solubility improvement near the isoelectric pH by increasing salt concentration in the extractant. Fractionation according to protein solubility in different solvents showed 13.97, 61.74, 0.48, 3.25 and 4.61 percent of albumin, globulin, prolamin, glutelin and insoluble protein, respectively. The in vitro digestibility determined by pepsin-pancreatin enzimatic sequence showed 85.59% and 90.54% of hydrolysis, in relation to casein, for the defatted flour and total globulin from baru seed, respectively. After 120 minutes of incubation, proteolysis of the major globulin obtained from Sephadex G-200 by tripsin, chymotrypsin and pepsin, separately, showed hydrolysis porcentages, in relation to casein, of 62.38, 17.92 and 71.38, respectively, and the progress of these proteolysis was monitored by SDS-PAGE. The chromatographic profile in gel filtration columns reveled the existence of one major protein form in total globulin, which was eluted off the DEAE-celulose ionexchange column at 0.12 M NaCl concentration. The molecular weight (MW) of globulin was estimated at 405-436 kDa and its electrophoretic profile showed three main bands of MW 62-64, 66 and 73-76 kDa, and... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2016-08-17T17:11:03Z (GMT). No. of bitstreams: 1 DISSERTACAO_MAYANNA KARLLA LIMA COSTA.pdf: 1242952 bytes, checksum: 59b9810ed129f3e0ae5fd5724a34dfe3 (MD5) Previous issue date: 2014-08-27 / The objective was to evaluate the nutritional value of varieties of sugar cane as a function of nitrogen fertilization. The varieties used were RB 863129, RB 867515 and RB 92579 and the nitrogen doses were 0, 60, 80, 120 and 140 kg ha-1. The experiment was divided in two stages: in the stage we evaluated the productivity and nutritional quality of three varieties of sugar cane (I Year - plant cane), and in the second stage (Years II and III) we evaluated the influence of nitrogen fertilization on the nutritional quality of the first and second ratoon of the same sugar cane varieties. For the analysis of productivity and other parameters, one collection of plant material was performed at the end of each experimental. The laboratory analysis consisted in the dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and (DMIVD) digestibility. In plant cane were also determinations of Brix and ratio NDF/Brix. For the plant cane, in terms of productivity in t ha-1, percentage of DM and CP, we observed a better values (P<0.05) for the RB 863129, and there was no significant difference between varieties (P <0.05) for the NDF, ADF and DMIVD. The three varieties evaluated showed reasonable chemical composition for ruminant nutrition and in vitro digestibility greater than 60%. The first and second ratoon all parameters differed (p<0.05) between varieties and fertilization, with the exception of DM content and in vitro digestibility occurring in most cases significant interaction between varieties, treatment and years of cutting. The variety RB 92579 presented at the two brass knuckles better productivity (a 131 t ha-1 of stalks) coupled with the best nutritional parameters (26% of ADF, 41% NDF, 2,44% CP and 69 DMIVD). The productivity of cane sugar, as well as its nutritional quality may suffer any increase or decrease according to the year and cutting management established in relation to nitrogen fertilization, moreover, the observed responses were influenced by varieties in most parameters assessed. / Objetivou-se avaliar o valor nutritivo de variedades de cana-de-açúcar em função da adubação nitrogenada. As variedades estudadas foram RB 863129, RB 867515 e RB 92579, e as doses nitrogenadas 0, 60, 80, 120 e 140 kg N ha-1. O experimento foi dividido em duas etapas, sendo que na primeira etapa avaliou-se a produtividade e qualidade nutricional de três variedades de cana-de-açúcar (Ano I - cana-planta), e na segunda, (Anos II e III) avaliou-se a influência da adubação nitrogenada na qualidade nutricional da primeira e segunda soca das mesmas variedades. Para a análise de produtividade e os demais parâmetros foram feitas três coletas de material vegetal, uma a cada final de ano experimental. Determinou-se os teores de matéria seca (MS), e, com base na MS, proteína bruta (PB), fibra em detergente neutro (FDN), fibra em detergente ácido (FDA) e digestibilidade in vitro da matéria seca (DIVMS). Na cana-planta foram determinados o teor de Brix e a relação FDN/Brix. Para a cana-planta, a produtividade (t ha-1) e os teores de MS e PB, foram superiores (P<0.05) para a variedade RB 863129. Não houve diferença (P<0,05) entre as variedades para os teores de FDN e FDA e para a DIVMS. As três variedades avaliadas apresentaram composição química razoável para nutrição de ruminantes e digestibilidade in vitro maior que 60%. Na primeira e segunda soqueira houve diferença (P<0,05) entre variedades e doses de adubação para os parâmetros avaliados, com exceção do teor de MS e digestibilidade in vitro, que foram influenciados (P<0,05) pelas variedades, doses de N e anos de corte. A variedade RB 92579 apresentou nas duas soqueiras a melhor produtividade (131 t ha-1 de colmos), aliada aos melhores parâmetros nutricionais (FDA 26%, FDN 41%, PB 2,4% e DIVMS 69,4%). A produtividade da cana-de-açúcar, assim com a sua qualidade nutricional, pode ser influenciada de acordo com o ano de corte e o manejo estabelecido em relação à adubação nitrogenada, além disso, as respostas observadas foram influenciadas pelas variedades na maioria dos parâmetros avaliados.

Actinobactérias são microrganismos capazes de produzir compostos bioativos presentes em extratos brutos com ação antibiótica e ionófora. A hipótese deste estudo foi de que extratos brutos de actinobactérias apresentem eficácia similar à monensina sódica para modulação da fermentação ruminal; no aumento da digestibilidade da matéria seca e matéria orgânica, aumento da concentração de propionato e redução da produção de gases totais, da desaminação protéica e de metano. Portanto, o objetivo foi avaliar in vitro dois extratos brutos (AMC e Caat) sobre a fermentação ruminal (produção total de gases, de ácidos graxos de cadeia curta (AGCC) e de metano (CH4); digestibilidade in vitro da matéria seca (DIVMS), da matéria orgânica (DIVMO) e produção de nitrogênio amoniacal (N-NH3). O estudo foi dividido em duas fases; a primeira objetivou estabelecer a curva dose-resposta dos extratos e a segunda para avaliação com controle positivo de monensina sódica. Para ambas fases, foram utilizadas como doadoras de líquido ruminal (inóculo) três vacas da raça Holandesa em lactação, portadoras de fístula ruminal. As dietas fornecidas foram à base de silagem de milho, farelo de soja, ureia, milho moído e suplemento mineral, com relação volumoso:concentrado de 60:40. Amostras de fluido ruminal foram coletadas por meio de bomba de vácuo, armazenadas em garrafas e caixas térmicas e transportadas ao laboratório para incubação em frascos de vidro. Para a primeira fase, foram avaliadas 4 doses (0,3; 0,6; 0,9 e 1,20 mg) de 2 extratos (AMC e Caat) em ensaio in vitro de 72 horas, no qual foram mensuradas a produção de gás durante a incubação e, ao final, as concentrações de AGCC, DIVMS e DIVMO foram determinadas. Na segunda fase, o ensaio foi de 24 horas, e ao final foram feitas análises para CH4 e N-NH3, além de AGCC, DIVMS, DIVMO e produção de gás. O delineamento experimental adotado foi o inteiramente casualizado (DIC) para ambas as fases. Na primeira fase, a inclusão do Caat diminuiu linearmente a DIVMO após 72 hs e reduziu as concentrações de ácido isovalérico, butírico, da relação acetato:propionato e da produção de gás acumulado por 24, 48 e 72 h. Na segunda fase, a inclusão dos extratos causaram diminuição da produção de gás acumulada em 24 horas, sendo menor no tratamento monensina e no Caat. Houve efeito de inclusão do Caat e da monensina sobre a concentração de AGCC, aumentando a concentração de propionato e diminuindo a concentração de butirato, acetato e a relação acetato:propionato. A concentração de N-NH3 foi menor mediante inclusão do extrato Caat e da monensina enquanto que a concentração de CH4 foi maior para a inclusão dos extratos quando comparados com a inclusão de monensina, entretanto não foram maiores do que o controle. A inclusão do extrato Caat foi capaz de modificar a fermentação in vitro de forma semelhante à monensina sódica, mas não alterou a digestibilidade da fibra. Por outro lado, o AMC não alterou a fermentação ruminal, seja na digestibilidade da fibra, na produção de gases totais, metano, nitrogênio amoniacal e AGCC / Actinobacteria are microorganisms that can produce bioactive compounds present in crude extracts with antibiotic and ionophore activity. Our hypothesis was that actinobacteria crude extracts showed similar efficacy to monensin on ruminal fermentation; increasing the digestibility of dry matter and organic matter, increasing propionate concentration and reducing the total gas production, protein deamination and methane. Therefore, the objective was to evaluate two crude extracts (AMC and Caat) on in vitro ruminal fermentation (total gas, short chain fatty acids (SCFA) and methane (CH4) production; in vitro digestibility of dry matter (IVDDM), of organic matter (IVOMD) and ammonia nitrogen production (NH3-N). The study was divided into two phases: the first aimed to establish the dose-response curve of the extracts and the second aimed to evaluate a positive monensin control. For both, it was used three lactating Holstein cows with ruminal fistula as rumen fluid donors (inoculum). The diets were based on corn silage, soybean meal, urea, ground corn and mineral supplement under forage:concentrate ratio of 60:40. Ruminal fluid samples were collected by a vacuum pump, stored in bottles and coolers and transported to the laboratory for incubation in glass bottles. For the first phase, they were evaluated 4 doses (0.3; 0.6; 0.9 and 1.20 mg) of 2 extracts (AMC and Caat) on a 72 hours in vitro assay, which were measured gas production during incubation and in the end, SCFA, IVDMD and IVOMD concentrations were determined. In the second phase, it was a 24 hours assay, and include final analyzes for CH4 and NH3-N, plus SCFA, IVDMD, IVOMD and gas production. The experimental design was completely randomized (DIC) for both phases. In the first phase, the Caat inclusion linearly decreased IVOMD after 72 hours and reduced isovaleric acid and butyric acid concentration and acetate:propionate ratio and cumulative gas production by 24, 48 and 72 h. In the second stage, the inclusion of the extracts caused a reduction of the cumulative gas production in 24 hours, being lower in monensin and Caat treatments. There was an effect of Caat and monensin inclusion on SCFA concentration, increasing the propionate concentration and reducing the butyrate and acetate concentration and the acetate: propionate ratio. The NH3-N concentration was lower by Caat inclusion and monensin while the CH4 concentration was higher for the extracts inclusion as compared with monensin, however were not bigger than control. The Caat inclusion was able to modify the in vitro fermentation similarly to monensin, but did not affect the fiber digestibility. Moreover, AMC did not affect the rumen fermentation, either in fiber digestibility, total gas, methane, ammonia nitrogen and SCFA production

Orientador: Flavia Maria Netto / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-14T13:49:46Z (GMT). No. of bitstreams: 1 Fernandes_MicheleAugusto_M.pdf: 2050778 bytes, checksum: aedc46d9b78bf413f7e34cfbe46a36a6 (MD5) Previous issue date: 2009 / Resumo: A ß-Lactoglubulina (ß-Lg) é uma das proteínas mais antigênicas presente no leite bovino. Tratamentos físicos, químicos ou enzimáticos podem alterar a antigenicidade desta proteína. Em trabalho anterior, verificou-se que o potencial antigênico da ß-Lg é reduzido quando polimerizada pela enzima transglutaminase (TG) na presença de cisteína (Cys). No entanto, o efeito da polimerização sobre o valor nutricional da ß-Lg ainda não é conhecido. O presente estudo teve como objetivo avaliar o efeito da reação de polimerização catalisada pela TG na digestibilidade in vitro e atividade antigênica da ß-Lg antes e após a ação das enzimas gastrintestinais. A ß-Lg polimerizada pela TG (0, 10 ou 25 U g-1), após tratamento térmico ou na presença de agentes redutores Cys (0, 0,1 e 0,25 mol L- 1) ou ditiotreitol (DTT 0,02 mol -1), foi avaliada quanto à digestibilidade in vitro, utilizando as enzimas pepsina e pancreatina. As amostras, antes e após a digestão in vitro, foram caracterizadas pelos métodos SDS-PAGE, SDSPAGE/ tricina e cromatografia líquida de alta eficiência de fase reversa (CLAE-FR). Posteriormente, as amostras foram avaliadas quanto à antigenicidade, exceto aquelas na presença de DTT, por meio do método de Imunoblote, utilizando soro de camundongos BALB/c sensibilizados com a ß-Lg na forma nativa (ß-Lg N) ou com ß-Lg polimerizada com 25 U de TG g-1, na presença de 0,25 mol L-1 de Cys (ß-Lg 0,25Cys 25TG). A adição de TG resultou na formação de polímeros com massa molar igual ou acima de 97,4 kDa, principalmente na presença de agentes redutores alcançando aproximadamente 96% de polimerização na presença de DTT e 91% na presença de Cys. A digestibilidade in vitro da ß-Lg N foi 53,6% e todos os tratamentos realizados aumentaram a digestibilidade da proteína em até 79%. Os maiores valores de digestibilidade foram obtidos quando a ß-Lg foi tratada com agentes redutores. O processo de polimerização também teve efeito positivo na digestibilidade, principalmente para as amostras polimerizadas na presença de Cys ou DTT, atingindo valores acima de 75%. A análise por Imunoblote mostrou que a polimerização da ß-Lg na presença de agente redutor Cys, na concentração de 0,25 mol L-1, reduziu o reconhecimento da ß-Lg pelas IgE específicas presente nos soros dos animais sensibilizados com ß-Lg N ou com ß- Lg 0,25Cys 25TG. Após a digestão com pepsina e pancreatina, as amostras polimerizadas pós-tratamento térmico ou na presença de Cys apresentaram redução da antigenicidade, como também os digeridos da ß-Lg tratada com Cys (não polimerizada com TG). A desnaturação pelo agente redutor Cys e a polimerização por TG em ambas as condições estudadas mostraram ser métodos efetivos no aumento da digestibilidade da ß-Lg. Por sua vez, a combinação destes métodos com a digestão por enzimas gastrintestinais levou à redução da antigenicidade da proteína, já que os peptídeos gerados apresentaram potencial antigênico baixo / Abstract: The ß-Lactoglubulin (ß-Lg) is one of the most antigenic proteins present in the bovine milk. Physical, chemical or enzymatic treatments can alter the antigenicity of this protein. In previous study, it was shown that the antigenic potential of ß-Lg is reduced when polymerized by transglutaminase (TG) in the presence of cysteine (Cys). However, the effect of polymerization on the nutritional properties of ß-Lg is still unknown. The present study aimed to evaluate the effect of the polymerization reaction catalyzed by TG on the in vitro digestibility and the antigenic activity of ß- Lg, before and after simulate digestion with gastrointestinal enzymes. The in vitro digestibility of the ß-Lg treated with TG (0, 10 or 25 U g-1), after heat treatment or in the presence of reducing agents Cys (0, 0.1 and 0.25 mol L-1) or dithiothreitol (DTT 0.02 mol L-1), was evaluated using the gastrointestinal enzymes, pepsin and pancreatin. The samples, before and after in vitro digestion, were characterized by SDS-PAGE, SDS-PAGE/tricine and reversed-phase high performance liquid chromatography (RP-HPLC). Subsequently, the samples were evaluated for antigenicity, except those prepared in the presence of DTT, by immunochemical methods (Immnoblotting), using sera from BALB/c mice sensitized with native ß-Lg (ß-Lg N) or ß-Lg polymerized with 25 U of TG g-1 in the presence of 0.25 mol L-1 of Cys (ß-Lg 0.25Cys 25TG). The formation of polymers with molar mass equal to or above 97.4 kDa was observed with the addition of TG, especially in the presence of reducing agents, reaching approximately 96% of polymerization in the presence of DTT and 91% in the presence of Cys. The in vitro digestibility of native ß-Lg was 53.6% and the all treatments performed increased the digestibility of protein up to 79%. The highest values of digestibility were obtained in the presence of reducing agents. The polymerization also had a positive effect on the digestibility, especially for those samples polymerized in the presence of Cys or DTT, with values above 75%. Analysis by immunoblotting showed that the polymerization of ß-Lg in the presence of 0.25 mol L-1 Cys, reduced the recognition of ß-Lg specific IgE present in the sera of animals sensitized with ß-Lg N or ß-Lg 0.25 Cys 25TG. After digestion with pepsin and pancreatin, the samples polymerized after heat treatment or in the presence of Cys showed reduced antigenicity. The digest of the samples treated with Cys (not treated with TG) was not recognized as antigens. The denaturation by Cys and polymerization by TG in both conditions were effective in increasing the digestibility of ß-Lg. In turn, the combination of these methods with digestion by gastrointestinal enzymes led to reduction of antigenicity of the protein and that peptides generated showed low antigenic potential / Mestrado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Mestre em Alimentos e Nutrição

Thesis (MScAgric (Animal Sciences))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: The purpose of this study was to investigate the variation that exists between maize samples regarding particle size separation, in vitro fermentation kinetics and in vitro dry matter (DM) disappearance. A second objective was to quantify possible relationships between the Roff Milling Index (RMI) of maize and any of the measured in vitro parameters. Three trials were conducted: a particle size distribution trial, a gas production trial and an in vitro DM degradability and starch disappearance trial. Overall, nine maize samples, which differed in terms of cultivar and endosperm type, were collected from different origins for the study. The samples were selected in terms of their Milling Index (MI). Three of the nine samples had a high MI that ranged between 109 and 118, three had a low MI that ranged between 67 and 71 and the other three samples had a medium MI that ranged between 85 and 92. Although the MI is not a direct indication of the hardness or softness of the endosperm, it was believed to be indirectly associated with vitreousness. In the first trial, the different maize samples were milled through a 1 mm screen and sieved through a series of three sieves 150, 125 and 106 μm, respectively. It was found that RMI was not a reliable indicator to predict particle size distribution, especially in terms of the coarse (>150 μm) and very fine (<106 μm) particles. In the gas production trial, the nine different maize samples were subjected to a gas production system for a duration of 48 hours. Here gas production and rate of gas production of the different maize types in buffered rumen liquor were measured during incubation. After fitting the gas volume data to the respective models, the non-linear parameters b, c and L were subjected to a main effects ANOVA with the aid of Statistica, version 9 (2009). Main effects were treatment and repetition. Means were separated by means of a Scheffé test and significance was declared at P < 0.05. The results were compared to the RMI of the different maize types and it was concluded that RMI was not a reliable predictor of gas production or rate of gas production of different maize types. In the third trial, in vitro DM degradability and starch disappearance of the different maize types were measured. In vitro DM degradability was conducted in the Ankom DAISYII incubator apparatus and the incubation times were 0, 2, 4, 8, 12 and 24 hours. Starch disappearance was measured on residues of the samples incubated for 0, 2 and 4 hours. After fitting the DM disappearance data to the respective models, the non-linear parameters a, b, c and L were subjected to a main effects ANOVA with the aid of Statistica, version 9 (2009). Main effects were treatment and repetition. Means were separated by means of a Scheffé test and significance was declared at P<0.05. The results indicated variation between maize samples, especially in terms of the a-, b- and L-values. The RMI did not appear to be a reliable predictor of digestibility parameters. / AFRIKAANSE OPSOMMING: Die doel van hierdie studie was om die variasie tussen mieliemonsters te ondersoek ten opsigte van die skeiding van partikelgroottes, in vitro-fermentasiekinetika en in vitro-droëmateriaalverdwyning. ‘n Tweede doel was om te bepaal of daar moontlike verwantskappe tussen die Roff Milling Index (RMI) van mielies en enige van die ander in vitro-parameters bestaan. Drie proewe is gedoen: verspreiding van partikelgrootte, ‘n gasproduksieproef en ‘n droëmateriaal degradeerbaarheid- en stysel verdwyningsproef. Nege mieliemonsters, wat van mekaar verskil ten opsigte van kultivar en endospermtipe, is van verskillende lokaliteite versamel. Die monsters is gekies in terme van hul maal-indeks (MI). Drie van die nege monsters het ‘n hoë MI gehad wat gewissel het tussen 109 en 118, drie het ‘n lae MI gehad wat gewissel het tussen 67 en 71 en die ander drie monsters het ‘n medium MI gehad wat gewissel het tussen 85 en 92. Alhoewel die MI waardes nie ‘n direkte indikasie van ‘n endosperm se hardheid- of sagtheidsgraad is nie, is dit aanvaar dat daar ‘n indirekte verwantskap tussen MI en glasagtigheid van die mielie bestaan. In die eerste proef is die nege verskillende mieliemonsters deur ‘n 1 mm sif gemaal en daarna deur ‘n reeks van drie siwwe met groottes van onderskeidelik 150, 125 en 106 μm gesif. Daar is bevind dat die RMI nie ‘n betroubare voorspeller is om partikelgrootte-verspreiding aan te dui nie, veral nie ten opsigte van growwe (> 150 μm) en baie fyn (< 106 μm) patikels nie. Tydens die gasproduksieproef is die nege mieliemonsters vir 48 ure blootgestel aan ‘n gasproduksiesisteem, waar gasdruk outomaties aangeteken is. Gasproduksie en tempo van gasproduksie van die verskillende mieliemonsters is gemeet en aangeteken gedurende inkubasie met ‘n gebufferde rumenvloeistofmedium. Nadat die gasvolumedata met behulp van relevante modelle gepas is, is die nie-linêre parameters b, c en L onderwerp aan ‘n hoof-effek ANOVA met die gebruik van Statistica weergawe 9 (2009). Hoof-effekte was behandeling en herhaling. Gemiddeldes is deur ‘n Scheffé-toets geskei en betekenisvolheid is verklaar by P<0.05. Die resultate verkry is vergelyk met die RMI van die verskillende mielietipes. Die gevolgtrekking is gemaak dat Roff MI nie ‘n betroubare voorspeller van totale gasproduksie of gasproduksietempo is nie. Tydens die derde proef is droëmateriaaldegradeerbaarheid en styselverdwyning van die verskillende mielietipes bepaal. In vitro droëmateriaal (DM) degradeerbaarheid is gedoen in die Ankom DAISYII inkubator met inkubasietye van 0, 2, 4, 8, 12 en 24 ure. Styselverdwyning is bepaal deur styselanalises op die residue van die monsters wat geïnkubeer is vir 0, 2 en 4 ure. Nadat die DM-degradeerbaarheid met behulp van relevante modelle gepas is, is die nie-lineêre parameters a, b, c en L onderwerp aan ‘n hoof-effek ANOVA met die gebruik van Statistica weergawe 9 (2009). Hoof-effekte was behandeling en herhaling. Gemiddeldes is deur ‘n Scheffé toets geskei en die betekenisvolheid is verklaar by P<0.05. Die resultate het aangedui dat daar groot variasie tussen mielies bestaan, veral ten opsigte van die a-, b- en L-waardes. Dit het verder geblyk dat die RMI van die verskillende mielietipes nie ‘n betroubare voorspeller van DM-degradeerbaarheid was nie.

The effects of oxidation by sodium hypochlorite (0, 0.8, 2, and 5%, NaOCl), the presence of endogenous proteins, and amylose content on waxy and common rice flours (WF, CF) and starches (WS, CS) were investigated in terms of in vitro starch digestibility, morphology and surface properties, and thermal and rheological characteristics. The concentration of NaOCl had an effect on all the samples including WF, CF, WS, and CS. The carbonyl and carboxyl group contents increased up to 25 and 10 folds (P < 0.05) of oxidized starches (WS, CS), respectively. Only mild oxidation (P < 0.05) occurred in flours (WF, WS). In addition, endogenous proteins were oxidized according to amino acid analysis and SDS–PAGE results. Glu+Gln, Gly, His, Arg, Tyr, and Lys were more sensitive to NaOCl oxidation. Disulfide bonds, hydrophobic force, and hydrogen bonds were involved in protein polymerization after NaOCl oxidative modification. In granular state, the in vitro starch digestibility of WF, WS, and CS decreased by 5% NaOCl oxidation. After gelatinization, only 2 and 5% oxidized WS had lower digestibility. Scanning electron microscopy and confocal laser scanning microscopy further demonstrated that protein existed on the surface of starch granules and had aggregation by oxidation. X-ray diffraction patterns showed the crystallinity of 5% oxidized flours and starches was reduced compared with all their non-oxidized samples. Thermal and rheological properties were analyzed by differential scanning calorimetry and rheometer, respectively. Starch gelatinization peak temperature of flours (WF, RF) was increased by 3 °C, but starches (WS, CS) had a significantly decrease by 8 °C. Viscoelastic patterns were dramatically changed by oxidation. Oxidized WF and CF had increased in both viscosity and elasticity by oxidation, whereas both WS and CS had significantly lower viscoelasticity after oxidative modification.

Orientador: Mauro dal Secco de Oliveira / Banca: Jane Maria Bertocco Ezequiel / Banca: Paulo de Figueiredo Vieira / Banca: Armando de Andrade Rodrigues / Banca: Maria Lúcia Pereira Lima / Resumo: Avaliou-se as digestibilidades in vitro da matéria seca (DIVMS), proteína bruta (DIVPB), fibra em detergente neutro (DIVFDN) e fibra em detergente ácido (DIVFDA), consumo de alimentos, produção e composição do leite e viabilidade econômica de dietas contendo 0, 20, 40, 60% de torta de girassol, em substituição ao farelo de girassol e milho, para vacas em lactação. As digestibilidades foram obtidas no fermentador ruminal DAISY II (Ankon® Technology), em um delineamento em blocos casualizados, com 4 tratamentos e 4 repetições. Para avaliar o consumo, produção e composição do leite, oito vacas da raça holandesa, com 113 dias em lactação, foram alimentadas com silagem de milho ad libitum e 1kg de concentrado para cada 3kg de leite produzido. Os animais foram distribuídos em dois quadrados latinos 4x4, primíparas e multíparas, com 4 períodos de 19 dias. Verificou-se que a inclusão de torta de girassol diminuiu linearmente (P<0,01) as DIVMS e DIVFDN em aproximadamente 0,014% para cada unidade percentual de torta adicionada e aumentou os consumos de extrato etéreo e mineral (P<0,01). O consumo de extrato etéreo foi 12,24% superior no tratamento com 60% de substituição comparado ao concentrado sem torta de girassol, mas não afetou o consumo total de matéria seca (13,55 kg/vaca/dia, em média). A porcentagem de proteína no leite diminuiu linearmente e a concentração de N-uréico no leite aumentou à medida que houve substituição do farelo pela torta de girassol. Houve aumento no custo por kg do concentrado. Dependendo da disponibilidade e conveniência econômica, a torta de girassol pode substituir até 60% do farelo de girassol. / Abstract: In vitro digestibility of dry matter (IVDDM), crude protein (IVDCP), neutral detergent fiber (IVDNDF) and acid detergent fiber (IVDADF), nutrients intake, milk production and composition, and economical viability of the diets with four levels of sunflower cake (0, 20%, 40%, 60%), in replacement to sunflower meal and corn grain for dairy cattle, was evaluated. The in vitro digestibilities of the concentrates were obtained through DAISY II Ruminal Fermentator (Ankon® Technology), in a randomized blocks design, with four treatments and fours repetitions. To evaluate the nutrients intake and milk production and composition, eight Holsteins cows, with 113 days in lactation, were fed with corn silage ad libitum and 1kg of concentrate for every 3kg of milk produced. The cows were divided in two Latin Square (4x4), primiparous and multiparous, in four periods of 19 days. It was found that the inclusion of sunflower cake decreased linearly (P<0.01) the IVDDM, IVDNDF, in 0,014% percentual unit and increased intake of mineral and ether extract (P<0.01). Ether extract intake was 12,24% higher for cows fed with 60% replacement compared with concentrate without sunflower cake, but not influenced the dry matter intake (13,55 kg/cow/day). The protein percentage decreased linearly and increased the N-urea concentration. Increase was observed in the cost by kilo of the concentrate as there was participation of the sunflower cake. Depending on the availability and economic convenience, the sunflower cake can substitute until 60% of the sunflower meal. / Doutor