Made available in DSpace on 2015-03-26T13:42:29Z (GMT). No. of bitstreams: 1 texto completo.pdf: 546340 bytes, checksum: c6ef1899560e5249142f8d7f9943743d (MD5) Previous issue date: 2006-09-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Soybean is one of the most important crops in the world extensively used as a food and feed source. However, the proteins present in soybean seeds are not considered ideal because they contain low amounts of the essential amino acids methionine and lysine. Adverse nutritional and other effects following consumption of raw soybean meal have been attributed to the presence of endogenous inhibitors of digestive enzymes and allergenic factors. Among these proteins are the P34 protein and the Bowman-Birk protease inhibitors. The absence of these factors would increase the nutritional quality of the soybean seeds, as well as the availability of this product to large number of consumers. Recent advances in plant biotechnology have enabled the silencing of specific genes. One of the techniques used to achieve this goal is RNA interference (RNAi). This method of posttranscriptional gene silencing is highly efficient, especially when the construct results in the formation of a intron-spliced hairpin RNA. It is proposed that RNAi works by enzymatic RNA degradation induced by double-stranded RNA. The main goal of this work was to construct expression cassettes for RNAi-based silencing of genes encoding antinutritional factors in soybean seeds. To achieve seed-specific expression, specific primers were designed for the amplification of the promoter region of the alpha-subunit of the beta-conglycinin gene. Appropriate restriction sites - Sac I and Xho I - were added to the primers which were used to amplify a 634 bp fragment. This product was cloned into a pGEMTEasy vector and cloning was verified by PCR, enzymatic digestion and sequencing. The sequenced clone was analyzed in silico for identification of cis-elements related to seed specific expression. The elements TATA box, CAAT box, RY LEG box and RY FLEB box were found. The fragment of interest was isolated from vector pGEM-TEasy by cleavage with enzymes Sac I and Xho I and inserted in the expression vector pKANNIBAL. This new vector was designated pBKN. For the construction of the expression cassettes, primers were designed according to published sequences deposited in the GenBank. The fragments of interest were isolated by the RT-PCR method using RNA isolated from soybean seeds. RT-PCR was also used to analyze the expression of the gene of interest in the seed and in other organs of the plant (root, stem and leaves). These analyses showed that the expression of these genes was continuous and abundant during all stages of seed development, and the corresponding transcripts were present in all organs analyzed. The fragments obtained by RT-PCR were sequenced and their identity was confirmed by BLAST analysis. To clone the sense sequence, the vectors pKANNIBAL and pBKN, and the fragments of interest were cleaved with enzymes Xho I and Kpn I. The ligation reaction was catalyzed by T4 DNA ligase and competent cells of Escherichia coli were transformed by heat shock. Cloning was verified by PCR and enzymatic digestion. The cloning of the antisense fragment was done using restriction enzymes Xba I and Cla I and vectors containing the sense fragments. The complete expression cassettes were cloned into the binary vector pCAMBIA 3301 and verified by PCR. / A soja é uma das mais importantes culturas do mundo e como apresenta um alto teor protéico, ela tem sido muito utilizada na alimentação animal e humana. Apesar disso, as proteínas encontradas na semente desta leguminosa não são consideradas ideais por apresentarem baixo teor de metionina e lisina, além de fatores alergênicos e de inibidores de proteases. Dentre estes fatores antinutricionais estão a proteína P34 e os inibidores de protease do tipo Bowman-Birk. A ausência destes fatores aumentaria a qualidade da semente de soja, bem como a tornaria disponível para uma faixa mais ampla de consumidores. Recentes avanços na biotecnologia vegetal têm possibilitado o silenciamento completo ou em altos níveis de genes específicos. Uma destas técnicas é conhecida como interferência por RNA ou RNA interference. Este método é altamente eficiente e a clivagem do mRNA alvo é induzida pela presença de pequenos RNAs dupla fita na célula. O objetivo deste trabalho foi a construção de cassetes de expressão para silenciamento gênico, via interferência por RNA, dos inibidores de protease do tipo Bowman-Birk e da proteína P34 em sementes de soja. Para a expressão semente-específica dos transgenes, primers específicos foram desenhados para a amplificação da região promotora do gene da subunidade [alfa] da [beta]-conglicinina. A estes oligonucleotídeos foram adicionados sítios de restrição Sac I e Xho I - adequados para sua clonagem em vetores de expressão em plantas. Foram clonados 634 pb no vetor pGEM-TEasy. A clonagem foi confirmada por PCR, clivagem enzimática e sequenciamento do clone isolado. A seqüência clonada foi analisada in silico para a identificação de cis-elementos relacionados à expressão semente específica de genes colocados sob controle deste promotor. Os elementos TATA box, CAAT box, RY LEG box e RY FLEB box encontrados. O fragmento de interesse foi, então, isolado do vetor pGEM-TEasy por clivagem com as enzimas Sac I e Xho I e inseridos em vetores de expressão em plantas (pKANNIBAL) originando o vetor pBKN. Para a construção dos cassetes de expressão, a escolha da região do cDNA de cada gene de interesse a ser amplificada foi determinada com o auxílio do programa BLOCK-iTTM RNAi Designer (INVITROGEN), e os primers específicos foram desenhados de acordo seqüências depositadas no banco de dados público GenBank. Inicialmente foi feita, por meio de RTPCR, uma análise da expressão dos genes de interesse durante o desenvolvimento da semente e em demais órgãos da planta (raiz, caule e folha). Essa análise evidenciou expressão contínua e abundante desses genes durante todo o desenvolvimento da semente bem como a presença de transcritos correspondentes em todos os órgãos vegetais analisados. Após sua amplificação, todos os fragmentos obtidos a partir de cDNA de semente foram sequenciados e sua identificação foi confirmada por meio de alinhamento utilizando o programa BLAST. Para a clonagem da seqüência sense, os vetores pKANNIBAL e pBKN, além dos fragmentos amplificados referentes aos genes de interesse, foram clivados com as enzimas Xho I e Kpn I. A reação de ligação foi realizada utilizando T4 DNA ligase e células de Escherichia coli ultracompetentes foram transformadas por meio de choque térmico. Os transformantes foram analisados por PCR e reação de clivagem enzimática. A clonagem do fragmento antisense foi realizada da mesma forma, utilizando as enzimas Xba I e Cla I e os clones contendo o fragmento sense como vetores. Em seguida, as construções foram transferidas para o vetor binário pCAMBIA 3301 e confirmadas por PCR.

O feijão é um importante alimento na dieta alimentar do brasileiro. A cada colheita ocorrem perdas dos grãos de feijão devido ao ataque de insetos e roedores. Uma das maneiras de se preservar os grãos de feijão, mantendo suas características nutricionais, é o uso da radiação gama. O objetivo do trabalho é avaliar os efeitos de diferentes doses da radiação gama nas propriedades físico-químicas do feijão carioca submetido à cocção e que sofreu ou não o processo de maceração. Partes dos feijões cariocas crus foram utilizadas como controle e outra parte foram submetidos à radiação ionizante com doses de 1, 5 e 10 kGy. Após a irradiação uma parcela das amostras (controle e irradiadas) passou pela maceração e outra não, em seguida, todos os feijões foram cozidos. As amostras cozidas foram liofilizadas, moídas e depois acondicionas em potes e armazenadas a temperaturas de -23ºC. As análises de composição centesimal, determinação da digestibilidade proteica, teores de tanino condensado e ácido fítico foram feitas utilizando as amostras moídas. Utilizando grãos inteiros, foram feitas análises de capacidade expansão e hidratação, tempo de cocção e cor instrumental. A irradiação não alterou a composição centesimal das amostras maceradas e sem macerar. Os teores de taninos condensados não reduziram conforme o aumento das doses. As concentrações de ácido fítico foram reduzidas nas doses de 5 e 10 kGy, para amostras maceradas, enquanto que para os feijões sem macerar, a irradiação gama não influenciou nos teores de fitatos. Ocorreu diminuição da digestibilidade proteica, nas amostras maceradas, a partir dose de 1 kGy, e as demais doses a redução não foi significativa. Já para os feijões sem macerar, houve aumento da digestibilidade, na dose de 10 kGy. Conforme o aumento das doses foi observado redução nos tempos de cocção dos feijões macerados e sem macerar. Na dose de 10 kGy, a capacidade de expansão do feijão aumentou. A coloração das amostras não modificou significativamente conforme o aumento das doses. Desta forma conclui-se que a radiação ionizante não influencia na composição centesimal e nos teores de taninos. A irradiação influenciou nos teores de ácido fítico em altas doses aplicadas, mas somente nas amostras maceradas, além de atenuar a digestibilidade proteica de feijões macerados. Reduz os tempos de cocção. Eleva a capacidade de expansão do feijão na dose de 10 kGy, porém não altera a coloração do feijão, conforme o aumento das doses / The common bean is an important component in the diet of the average Brazilian person. Each harvest of beans, losses occur due to attacks of insects and rodents. One of the ways to preserve the beans, and at the same time keep its nutritional characteristics, is the use of gamma radiation. This study aimed to evaluate the effects of different doses of gamma radiation on the physical and chemical properties of the carioca variety of common beans subject to cooking of soaked and unsoaked beans. Portions of raw beans were used as control and the other was subject to ionizing radiation at doses of 1, 5 and 10 kGy. Following irradiation, a portion of the samples (control and irradiated) were soaked and the other was not, then all beans were cooked. The cooked samples were lyophilized, milled and then accommodated in pots and stored at -23ºC temperatures. The analysis of chemical composition, determination of protein digestibility, condensed tannin and phytic acid content were performed using the milled samples. Using whole grains, were performed analysis of expansion capability and hydration, cooking time and instrumental color. Irradiation did not alter the chemical composition of soaked and unsoaked samples. The condensed tannin levels did not reduce according to increased doses. The phytic acid concentrations were reduced at the doses of 5 and 10 kGy for soaked samples, whereas for the not soaked beans, gamma irradiation did not influence the phytate content. The protein digestibility decreased on soaked samples, at doses of 1 kGy and in the other doses, the reduction was not significant. As for not soaked beans, increases in digestibility were observed at dose of 10 kGy. As the doses increased, reduction in cooking time on soaked and unsoaked beans was noted. At a dose of 10 kGy, the bean expansion capability increased. The samples\' color did not change significantly, as the doses increased. Therefore, it is concluded that ionizing radiation has no effect on the chemical composition and tannin content. Irradiation influenced the phytates when applied in higher doses, but only in soaked samples, as well as mitigate protein digestibility on soaked beans. It reduces cooking time, raises bean expansion capability at a 10 kGy dose, but it does not change the bean color as doses increase

Dietary protein quality depends on two key measures: true ileal protein digestibility and the metabolic utilization of absorbed amino acids (AA). The objectives of this study were to determine the influence of two dietary factors (antinutritional factors and peptides) on ileal endogenous protein flows; to validate the intubation technique used in humans for ileal digesta sampling; to determine the postprandial metabolic utilization of dietary AA depending on their delivery form and to assess the validity of the growing pig for predicting true ileal protein digestibility in the adult human. Investigations were undertaken in the growing rat, growing pig and adult human. Ileal digesta were collected from euthanised rats, post valve T-caecum cannulated pigs, and naso-ileal intubated conscious adult humans. Ileal endogenous nitrogen (N) and AA were measured using a protein-free (PF) diet, diets containing 15N-labelled casein in the intact (C) or hydrolysed (HC) form, or a diet based on free AA (diet A), for which some dispensable AA were omitted to allow a direct determination of their endogenous flows. Digesta centrifugation and ultrafiltration (diet HC) allowed for the determination of ileal endogenous protein flows and the extent of tracer (15N) recycling. Antinutritional factors from a crude extract of kidney beans (Phaseolus vulgaris), when given at amounts commonly ingested in practice, enhanced ileal endogenous protein flows (rats, PF diet). After adaptation to the diet, body N balance per se did not influence ileal endogenous protein flows (rats, diets PF and A) but dietary peptides led to greater ileal endogenous AA and N flows compared with a protein-free diet. Dietary peptides (HC), compared with peptides naturally released in the gut during protein digestion (C), did not enhance ileal endogenous protein flows (rats, pigs, and humans). The extent of tracer recycling, however, was maximal in frequently-fed rats, lower in meal-fed pigs and minimal in meal-fed humans (65, 21, and 11% of 15N-labelled ileal endogenous proteins, respectively). Naso-ileal intubation for ileal digesta sampling in humans was shown to be an accurate method and evidence was obtained supporting the growing pig as a valid model for predicting true ileal protein digestibility in the adult human. Finally, the form of delivery of dietary AA (from HC or C) influenced the postprandial metabolic fate of dietary AA, especially in terms of AA catabolism kinetics. However, the overall nutritional value of C and HC were similar.

The current general economic and food crises have generated an unsettled future for food and feed production and prices in general. Increasing demand, prices and fluctuations in supply in world markets for fishmeal, fish oil, soybean meal, maize and wheat meal emphasize the need to reduce the dependence of the fish feed industry on these ingredients by increasing choices among a wider range of raw materials. Legume seed such as peas, chickpeas and faba beans are promising ingredients for aquafeeds due to their high protein compared to cereals but also for their energy content. The objective of the present study was to investigate the use of these legumes in both high and low inclusion level in diets for the two main species farmed in the Mediterranean countries namely, European seabass (Dicentrarchus labrax L.) and gilthead seabream (Sparus aurata L.). In Chapter 3 the effects of different processing conditions were examined on whole seed flours of tested ingredients with respect to both nutritional and antinutritional factors and physical characteristics of the experimental diet pellets including high and low levels of each legume. Extrusion of raw material resulted in a clear reduction in trypsin inhibitors (TI) with chickpeas showing a decrease of up to 90% and complete inactivation for wheat flour, while for peas and faba beans reduction was less than 50% in most cases. Extrusion processing was less effective in the reduction of phytic acid and total tannins, occasionally reaching 22% and 18% respectively. Oligosaccharides and total NSP were not drastically affected by processing, however faba bean NSP showed greater reduction. A redistribution of soluble and insoluble NSP fractions was noted for chickpea and pea flours after extrusion. Physical characteristics of the pellets were not negatively affected for any of the tested diets. In Experiment I the effects of total or partial wheat substitution by legumes were investigated on nutrient digestibility, gastrointestinal evacuation rate and serum glucose response of European seabass. Use of legumes in seabass and seabream diets resulted in an overall increase in gastrointestinal evacuation time and a delay in glucose load. Specifically, gastric evacuation time was greatly delayed when seabass was fed a diet with high levels (30%) of chickpeas, while foregut evacuation time was mostly delayed by the diet including a high level (30%) of faba beans. In addition, glucose levels in seabass serum were also affected by the type of carbohydrates ingested with wheat starch showing more rapid increase and decrease of glucose compared to fish fed pea and chickpea diets, while faba bean starch resulted in a lower glucose peak. In Experiment II growth, digestibility, hematological parameters, histological effects and fillet organoleptic characteristics and the interaction between starch inclusion level (8% and 16% or 17% and 35% of legumes respectively) and legume type were estimated when tested legumes replaced wheat in European seabass diets. Digestibility coefficients were satisfactory for all nutrients (>93%) while legume diets at a low level had beneficial effects on growth parameters when compared to the control diet, with chickpeas showing a significant improvement in SGR (P<0.05). High level legume diets did not result in any negative effect on growth. HSI was increased with increasing starch/legume inclusion in the feed and serum glucose also increased for fish fed high levels of faba beans and chickpeas. Carcass proximate composition was not affected by replacement of wheat in the diets, excluding the increase of fat content in fish fed chickpeas. Sensory analysis showed no differences between fish fed the control and high legume inclusion diets. Lastly in Experiment III growth, hematological parameters, histological effects and the interaction between starch inclusion level (low and high) and legume type were evaluated when tested legumes were included in gilthead seabream diets. Decreased, but not significantly so, growth was observed for all diets including legumes compared to the control. Poorer SGR were observed for pea and faba bean diets when these legumes were included at high levels. Liver glycogen increased with increasing starch level, but HSI did not differ significantly for any of the diet treatments. Histological examination of hindgut did not show pathological effects, such as enteritis, for in either species or for any of the diets. Increased absorptive vacuoles were found for control and pea diets (high level) only for seabass. The findings of this thesis showed that the two important species cultivated in Mediterranean countries responded differently to the same raw materials used at high levels in the diets. Overall legumes had a strong effect on gastrointestinal evacuation reducing the rate of feed or digesta passage. Peas, chickpeas and faba beans successfully replaced wheat in seabass diets resulting in improved growth coefficients. However, when the same legumes included in seabream diets growth performance was not improved compared to the wheat based diet.

Darbo tikslas: ištirti kvietrugių maistinę vertę ir jų panaudojimo galimybes viščiukų broilerių mityboje. Darbo uždaviniai: ištirti kvietrugių maistinę vertę; ištirti kvietrugių įtaką viščiukų broilerių virškinimo procesams. Rezultatai: išanalizavus 11 kvietrugių veislių nustatytas vidutinis žalių baltymų kiekis – 11,99 proc., žalių riebalų – 0,83 proc., žalių pelenų – 1,31 proc., vidutinis krakmolo kiekis – 69,69 proc., kalcio – 0,07 proc., fosforo – 0,303 proc. Pagrindinių aminorūgščių, lizino, metionino, treonino ir triptofano vidutiniai kiekiai atitinkamai nustatyti – 0,36 proc., 0,17 proc., 0,3 proc., 0,13 proc. Naudojant viščiukų lesinimui multifunkcinį fermentą bei kvietrugių veislę Tulus žarnyno svoris, ilgis bei aklosios žarnos ilgis mažėjo 4 – 6 proc., o naudojant veislę Lego - esminių pokyčių nenustatyta. Naudojant fermentą bei kvietrugių veisles Lego ir Tulus širdies svoris padidėjo atitinkamai 8 ir 10 proc., lesinant minėtas kvietrugių veisles ir fermentą kepenų svoris padidėjo atitinkamai 10 ir 8 proc. Naudojant viščiukų lesinimui multifunkcinį fermentinį priedą virškinamosios masės klampumas viščiukų broilerių aklojoje žarnoje padidėjo 11 – 23 proc. / Purpose of work: investigate the nutritional value of triticale as well as the use of it as a feed to broiler chicken. The tasks of work: investigation of triticale nutritional value; investigation of triticale influence to digestive processes of broiler chicken. Results: an analysis of 11 triticale varieties has shown that the average crude protein content is 11.99 percent, crude fat 0.83 percent, green ash 1.31 percent, starch 69.69 percent, calcium - 0.07 percent, phosphorus - 0.303 percent. The average amount of basic amino acids, lysine, methionine and tryptophan in triticale are as follows: 0.36 percent, 0.17 percent, 0.3 percent, 0.13 percent. The research has shown that by using multifunctional enzymes and triticale variety Tulus intestinal weight, length and caecum length decreased by 4-6 percent; whereas the use of triticale variety Lego has resulted in no major differences. The use of both varieties Lego and Tulus resulted in increase of heart weight by 8-10 percent and increase in liver weight by 10 for Lego and 8 percent for Tulus variety. The use of multifunctional enzyme has also resulted in viscosity of digestible mass in an increase in broiler caecum by 11-23 percent.

Orientadora: Margarida Maria Barros / Banca: Dalton José Carneiro / Banca: Edma Carvalho de Miranda / Resumo: A pesquisa teve por objetivo avaliar o coeficiente de digestibilidade aparente (CDA) da soja crua (SC) e a substituição (0,0; 15,0; 25,0; 35,0%) da proteína do farelo de soja pela proteína da soja crua em dietas práticas, sobre o desempenho produtivo, resposta hematológica e análise histopatológica do fígado. A pesquisa foi dividida em dois estudos: Estudo I - ensaio de digestibilidade. Foram utilizados 80 juvenis (± 50,0 g) de tilápia-do-nilo num sistema para determinação dos CDA, sendo a ração teste acrescida de 0,1% de cr2o3. Estudo II - ensaio de desempenho produtivo e saúde. Por 90 dias. Foram utilizados 160 peixes com peso médio inicial de 17,0 ± 1,55 g, distribuídos em 32 tanques-rede de 200L (cinco peixes/aquário). Foi utilizado um delineamento inteiramente casualizado, com quatro tratamentos e oito repetições. Ao final do período experimental determinou-se o desempenho produtivo (ganho de peso, consumo de ração, conversão alimentar aparente, taxa de eficiência proteica, taxa de retenção proteica e porcentagem de sobrevivência). Posteriormente, foram efetuadas as análises hematológicas dos peixes (contagem de eritrócitos, porcentagem de hematócrito, taxa de hemoglobina e contagem diferencial de leucócitos) e aos 45 e 90 dias as lâminas histológicas dos fígados. Os peixes alimentados com ração sem SC apresentaram melhor ganho de peso e consumo de ração. Sinais de estresse, confrontos agonísticos, dominância entre os animais e lesão dérmica foi observado nos peixes alimentados com rações contendo níveis crescentes de soja crua. O fígado dos peixes não apresentou edema ou necrose, apenas degeneração. A soja crua prejudicou o desempenho produtivo da tilápia-do-nilo, porém no período de 90 dias, não determinou alterações no eritrograma e histologia do fígado / Abstract: The research aimed to evaluate the apparent digestibility coefficient (ADC) of raw soybean (RS) and the replacement of protein from soybean meal by raw soybean protein in increasing levels of RS (0.0, 15.0, 25 , 0, 35.0%) in practical diets on productive performance, hematological response and histopatological analysis of liver. The present research was divided into two studies: Study I - the digestibility trial. In this study 80 juveniles (± 50.0 g) of Nile tilapia were used to determining the ADC. Chromic oxid was added in test diet at a rate of 0.1%. Study II - productive performance and health trial. In this study the experimental period was 90 days. One hundred and sixty fish with initial average weight of 17.0 ± 1.55 g were used. The animals were distributed into 32 cages of 200L containing five fish per aquarium. Treatments were arranged in a completely randomized design with four treatments and eight replicates. At the end of the experimental period fish were weighed to determine productive performance (weight gain, feed intake, apparent feed conversion, efficiency protein ratio, protein retention ratio and survival percentage). Hematological parameters evaluated after 90 days were total (erythrocyte and leukocytes count, differential leucocytes, hematocrit, hemoglobin, total plasmatic protein, albumin and globulin). Hematimetric indices were calculated. At the 45th and the 90th days histological slides of fish livers were performed. Fish fed diet with no RS showed better weight gain and feed intake. Signs of stress, agonistic behaviors, dominance among animals and skin lesions were observed in fish fed diets containing raw soybean. Fish livers showed no edema or necrosis, only degeneration. Raw soybean impaired growth performance, however 90 days did not determined alterations neither on erythrogram nor in histological characteristics of liver / Mestre

Resumo: Várias oleaginosas estão sendo estudadas para a produção de biocombustíveis. O processamento do grão para a obtenção do óleo resulta numa diversidade de subprodutos, entre eles o farelo de nabo forrageiro (Raphanus sativus) que, por possuir alto teor protéico, se apresenta como possível sucedâneo do farelo de soja nas rações. Entretanto, faltam informações de seu valor nutritivo e da ação dos fatores antinutricionais presentes. Este estudo foi realizado na Unesp - Universidade Estadual Paulista, da Faculdade de Medicina Veterinária e Zootecnia, Campus de Botucatu, para determinar os coeficientes de digestibilidade aparente (CDA) da matéria seca (MS), proteína bruta (PB), energia bruta (EB), aminoácidos (AA) e a disponibilidade do fósforo do farelo de nabo forrageiro, pela tilápia-do-Nilo. Empregouse uma ração purificada (referência) e uma ração composta de 60% da ração referência e 40% do farelo de nabo forrageiro. O farelo de nabo forrageiro apresentou 91,28% de MS; 42,07% de PB, 4256 kcal/kg de EB, 3,47% de EE, 7,37% de FB, 1,25% de cálcio e 1,0% de fósforo. Os CDA foram de 55,92% para MS, 82,10% para PB, 75,26% para EB e 85,23% do fósforo. Os CDA dos AA estiveram entre 81,12% para a glicina e 95,11% para o ácido glutâmico. Os resultados demonstram que o farelo de nabo forrageiro apresenta potencial para ser usado como fonte protéica alternativa para compor a ração da tilápia do Nilo. / Abstract: Oil seeds have been studied to produce bio combustibles. In order to obtain oil, seed processing results in several byproducts, among them, the fodder radish meal (Raphanus sativus) which has high protein content. This byproduct is a potential substitute to soybean meal in feed manufacture. However, there is a lack of information regarding its nutritional value and the anti-nutritional content and effects. This study was conduced at the Veterinary Medicine and Animal Science College from Sao Paulo State University, Botucatu, to determine the apparent digestibility coefficient (ADC) of dry matter (DM), crude protein (CP), raw energy (RE), amino acids (A) and phosphorus availability. The fish were fed a purified feed (base diet) and a 60% base diet plus 40% of fodder radish meal feed. Radish meal presented 91.98% DM, 42% CP, 4256 kcal/kg RE, 3.47% Ether extract, 7.37% crude fiber, 1.25% calcium and 1.0% phosphorus. The ADC values were DM: 55.92%, CP: 82.10%, RE: 75.26% and phosphorus: 85.23%. The ADC value for AA was 81.12% for glycine and 95.11% for glutamic acid. The results presented a potential utility for radish meal as an alternative protein source for Nile tilapia feeds. / Orientador: Luiz Edivaldo Pezzato / Coorientador: Margarida Maria Barros / Banca: Wilson Maddamitu Furuya / Banca: José Roberto Sartori / Mestre

Submitted by Irene Nascimento (irene.kessia@ufpe.br) on 2016-07-08T17:05:49Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Bernardo do Rego Belmonte_Mestrado.pdf: 1797925 bytes, checksum: 7816587bf6318d594398ab8d0d619d73 (MD5) / Made available in DSpace on 2016-07-08T17:05:49Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Bernardo do Rego Belmonte_Mestrado.pdf: 1797925 bytes, checksum: 7816587bf6318d594398ab8d0d619d73 (MD5) Previous issue date: 2015-07-30 / FACEPE / Lectinas são proteínas que apresentam atividade inseticida, sendo capazes de interferir na alimentação, desenvolvimento, reprodução e sobrevivência de insetos. Sitophilus zeamais, conhecido como gorgulho-do-milho, destaca-se como uma das principais pragas de grãos armazenados no Brasil e causa danos aos grãos de milho, trigo, sorgo e arroz. Myracrodruon urundeuva (aroeira-do-sertão) é uma árvore, madeira de lei, amplamente distribuída no Brasil. O presente trabalho avaliou os efeitos deletérios de extratos e lectinas isoladas da entrecasca (MuBL) e do cerne (MuHL) de M. urundeuva sobre o gorgulho-do-milho. Para determinação da atividade inseticida, a amostra foi incorporada a discos de farinha de trigo que serviram como dieta para os insetos durante 7 dias. As faixas de concentração testadas foram: 4 a 20 mg do extrato da entrecasca por g de farinha de trigo, 14 a 70 mg/g (extrato do cerne), 0,6 a 3,0 mg/g (MuBL) e 1,4 a 7,0 mg/g (MuHL). Após o término do bioensaio, foi avaliada a taxa de mortalidade e os seguintes parâmetros nutricionais: índice de deterrência alimentar (IDA), taxa de consumo relativo (TCR), taxa de ganho relativo de biomassa (TGB) e eficiência na conversão do alimento ingerido (ECAI). Em seguida, foram realizados ensaios com o extrato da entrecasca, extrato do cerne, MuBL e MuHL em concentrações de 20 mg/g, 70 mg/g, 3 mg/g e 7 mg/g, respectivamente, e após 7 dias, extratos de intestino dos insetos foram obtidos e avaliados quanto às atividades de protease, celulases (endoglucanase, exoglucanase, β-glicosidase), fosfatases (ácida e alcalina) e α-amilase. As dietas contendo os extratos de entrecasca e do cerne não induziram mortalidade dos insetos em um período de 7 dias de experimento, bem como não exerceram efeito deterrente sobre os insetos. Contudo, ambos os extratos interferiram nos parâmetros nutricionais dos insetos. Em todos os tratamentos com os extratos houve redução da biomassa corporal, uma vez que os valores de TGB foram negativos. Os valores de ECAI também foram negativos, variando de -3% a -70%. Com relação à TCR, os valores não foram significativamente diferentes (p > 0,05) que no controle, corroborando com a ausência de efeito deterrente. A comparação entre as concentrações testadas e os valores obtidos para os parâmetros nutricionais revela que o extrato da casca foi mais efetivo que o extrato do cerne. MuBL não induziu mortalidade em 7 dias, mas afetou a incorporação da dieta pelos insetos, como evidenciado pelos valores negativos de TGB e ECAI. Os valores de TCR foram maiores que no grupo controle, indicando a ausência de efeito deterrente. MuHL apresentou efeito deterrente variando de 44,82% (fraca) a 89,65% (forte) de acordo com a concentração e afetou fortemente a nutrição dos insetos, com valores de ECAI chegando a -360%. Uma vez que todas as amostras testadas apresentaram efeitos antinutricionais, as atividades de enzimas digestivas em insetos do grupo controle e que ingeriram os extratos e as lectinas foram avaliadas. A ingestão do extrato da entrecasca não resultou em alteração significativa (p > 0,05) das atividades de protease, fosfatase ácida, fosfatase alcalina e β-glicosidase. Já as atividades de endoglucanase, exoglucanase e α-amilase foram maiores em relação ao controle. Os insetos que ingeriram o extrato do cerne apresentaram atividades de protease, endoglucanase, exoglucanase e α-amilase menores em comparação ao controle enquanto a atividade de fosfatase ácida foi maior. No tratamento com MuBL, as atividades de fosfatase ácida e α-amilase foram menores em relação ao controle. Por outro lado, as atividades de protease, endoglucanase e exoglucanase foram maiores. No tratamento com MuHL, as atividades de fosfatase alcalina, fosfatase ácida e exoglucanase foram menores em relação ao controle. Em conclusão, extratos e lectinas de entrecasca e cerne de M. urundeuva exerceram efeitos antinutricionais sobre adultos de S. zeamais, apresentando potencial para uso no controle dos danos causados por esse inseto-praga. / Lectins are proteins with insecticidal activity able to interfere with the feeding, development, reproduction and survival of insects. Sitophilus zeamais, known as maize weevil, stands out as one of the main stored grain pests in Brazil and causes damage to maize, wheat, sorghum and rice. Myracrodruon urundeuva (aroeira-do-sertão) is a hardwood tree broadly distributed in Brazil. The present work evaluated the deleterious effects of extracts and lectins isolated from the bark (MuBL) and heartwood (MuHL) of M. urundeuva against the maize weevil. For determination of insecticidal activity, the sample was incorporated into wheat flour disks that served as diet for the insects during 7 days. The concentration ranges tested were: 4–20 mg of bark extract per gram of wheat flour, 14–70 mg/g (heartwood extract), 0.6-3.0 mg/g (MuBL) and 1.4-7.0 mg/g (MuHL). After the end of bioassay, it was evaluated the mortality rate and the following nutritional parameters: feeding deterrence index (FDI), relative consumption rate (RCR), relative biomass gain rate (BGR) and efficiency in conversion of ingested food (ECIF). Next, it was performed assays with the bark extract, heartwood extract, MuBL and MuHL at concentrations of 20 mg/g, 70 mg/g, 3 mg/g and 7 mg/g, respectively, and after 7 days, extracts from insects gut were obtained and evaluated for the activities of protease, cellulases (endoglucanase, exoglucanase, β-glucosidase), phosphatases (acid and alkaline) and α-amylase. The diets containing the bark and heartwood extracts did not induce mortality of the insects in a period of 7 days of assay as well as did not exert deterrent effect on the insects. However, both extracts interfered with the nutritional parameters of the insects. In all treatments with the extracts, there was a reduction of body biomass since the values of BGR were negative. The values of ECIF were also negative, ranging from -3% to -70%. Concerning RCR, the values were not significantly (p > 0.05) from control, corroborating the absence of deterrent effect. The comparison between the tested concentrations and the values obtained for the nutritional parameters reveal that the bark extract was more effective than the heartwood extract. MuBL did not induce mortality in 7 days but affected the incorporation of the diet by the insects, as evidenced by the negative values of BGR and ECIF. The RCR values were higher than in control group indicating the absence of deterrent effect. MuHL showed deterrent effect ranging from 44.82% (weak) to 89.65% (strong) according to the concentration and affected strongly the insect nutrition, with ECIF values reaching -360%. Since all the tested samples showed antinutritional effects, the activities of digestive enzymes from insects of control group and that ingested the extracts and lectins were evaluated. The ingestion of bark extract did not result in significant alteration (p > 0.05) of the activities of protease, acid and alkaline phosphatases and β-glucosidase. However, the activities of endoglucanase, exoglucanase and α-amylase were higher concerning control. The insects that ingested the heartwood extract showed protease, endoglucanase, exoglucanase and α-amylase activities lower than in control while the activity of acid phosphatase was higher. In treatment with MuBL, the acid phosphatase and α-amylase activities were lower than in control. On the other hand, the activities of protease, endoglucanase and exoglucanase were higher. In the treatment with MuHL, the activities of alkaline and acid phosphatases and of exoglucanase were lower than in control. In conclusion, the extracts and lectins from bark and heartwood of M. urundeuva exerted antinutritional effects on S. zeamais adults, showing potential for use in control of the damages caused by this insect pest.

Orientador: Jaime Amaya-Farfan / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-08T21:26:36Z (GMT). No. of bitstreams: 1 BarruetoGonzalez_NorkaBeatriz_D.pdf: 1348327 bytes, checksum: 18f0856e8caba5f78415b8cd169e869a (MD5) Previous issue date: 2007 / Resumo: Dada a importância que os feijões secos têm na dieta tradicional brasileira, a Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) tem desenvolvido novos cultivares de feijão-comum (P. vulgaris), por meio melhoramento genético, para atender os altos padrões agronômicos e culinários do mercado interno. Entretanto, as características nutricionais destas novas cultivares estão documentadas de maneira incompleta, tanto em termos de conteúdo de macro/micro nutrientes, como também de seus valores biológicos. O objetivo do presente trabalho foi avaliar o valor nutritivo de sete novas cultivares de feijão: Jalo Precoce, Radiante, Vereda, Pérola, Timbó, Valente e Ouro Branco, da EMBRAPA do Centro de Pesquisa Arroz-Feijão, Goiás. Após análise centesimal de rotina, foram determinados o perfil aminoacídico e alguns importantes componentes minerais (cálcio, ferro, magnésio, cobre e zinco). Os resultados mostram que variações significativas (P<0,05) ocorreram no conteúdo protéico e de carboidratos totais entre todas as cultivares. O teor de proteína variou entre 21 e 28%, com destaque para Radiante, Jalo Precoce e Ouro Branco que tiveram as maiores concentrações. O teor de carboidratos foi de 55,4 (Radiante) a 62,1% (Valente), entretanto, os lipídios permaneceram constantes, variando de 2,4 a 2,8%. As proteínas de todas as novas cultivares alcançaram os padrões internacionais para os requerimento de histidina, valina, isoleucina, leucina, fenilalanina e tirosina. Com destaque para as cultivares Vereda e Ouro Branco, que alcançaram os requerimentos de lisina e treonina. Por outro lado, todas as cultivares apresentaram baixo conteúdo de aminoácidos sulfurados, metionina e cistina. Não foram encontradas diferenças estatísticas significativas entre as cultivares em termos de Mg, Fe e Zn, mas as concentrações de Ca e Cu variaram significativamente (P<0,0001). O conteúdo de ferro foi de 4,7 (Ouro Branco) a 5,5mg/100g (Vereda e Valente), enquanto que o cobre foi de 0,5 (Ouro Branco) a 0,9mg/100g (Radiante), e o conteúdo de zinco foi de 2,4 (Ouro Branco e Pérola) a 3,1mg/100g (Jalo Precoce). Pelos altos teores de Ca se destacaram as variedades Pérola (486), Vereda (428) e Valente (387mg/100g), enquanto que Jalo Precoce, Pérola e Valente apresentaram os maiores teores de Mg (129-117mg/100g). Considerando o fato de que todas as variedades foram cultivadas sob condições estritamente controladas de solo, água e práticas agrícolas, estes resultados podem ser altamente representativos com relação às características individuais de composição, onde as variedades Jalo Precoce, Ouro Branco e Valente apresentaram-se mais vantajosas em termos de composição de nutrientes. A partir destes resultados, três novas cultivares de feijão-comum (P. vulgaris) foram selecionadas, por causa do seu conteúdo de nutrientes, para avaliação da biodisponibilidade mineral e comparar com um cultivar clássico de soja (Glycine Max), var. Conquista. As três cultivares foram: feijão branco ¿Ouro Branco¿, feijão preto ¿Valente¿ e Carioca ¿Pérola¿ e os minerais testados foram os previamente estudados: cálcio, magnésio, cobre e zinco. O balanço metabólico, coeficiente de digestibilidade aparente (CDA) e a taxa de retenção e absorção (R/A) para estes minerais foram os parâmetros determinados, usando ratos Wistars recém-desmamados. Os animais receberam por doze dias dietas contendo exclusivamente os feijões cozidos ou farinha integral de soja tostada. Os níveis de fitatos foram determinados pelo método colorimétrico de Latta & Eskin e os minerais por absorção atômica nas dietas, fezes, urina e fêmur. Além disso e com o objetivo de avaliar o impacto das dietas no crescimento do osso cortical das tíbias, foram feitas medidas histomorfométricas da altura e espessura da cartilagem. Os resultados mostraram que as dietas à base de leguminosas diferiram estatisticamente uma das outras em termos de cálcio (275-480mg/100g), magnésio (108-262mg/100g), cobre (0,5-1,06mg/100g) e zinco (2,4-5,0mg/100g), e que as dietas à base de feijão Carioca ¿Pérola¿ e feijão preto ¿Valente¿, bem como a soja foram as que maior conteúdo mineral apresentaram. As dietas à base de feijão mostraram baixos teores de fitatos (3,1-3,9mg/g), especialmente com relação à soja (6,5mg/g). A análise estatística do balanço mineral detectou diferenças significativas entre os índices biológicos obtidos para os tratamentos de feijão e soja. As dietas ¿Valente¿ e ¿Pérola¿ mostraram alto balanço de cálcio (22-25mg/d), em comparação com as dietas ¿Ouro Branco¿ (12,8mg/d) e soja (14,4mg/d). A taxa de retenção e absorção (R/A), entretanto, foi maior para a dieta soja (99,0%), seguida pela dieta ¿Valente¿ (96,7%). Com relação ao CDA para cálcio, a dieta ¿Pérola¿ se destacou (92,5%) entre todos os tratamentos à base de leguminosas e Controle (AIN-93G). Os índices de avaliação revelaram que a soja teve maior balanço de magnésio (4,7mg/d) entre todas as dietas è base de feijão, entretanto o CDA foi melhor para a dieta ¿Pérola¿ (82%), apesar da taxa R/A ter sido melhor para ¿Valente¿ (7,1%) e soja (6,8%), enquanto que ¿Ouro Branco¿ e Controle (AIN-93G) tiveram índices negativos (valores de balanço e R/A). Com relação ao cobre, todas as dietas à base de leguminosas tiveram resultados negativos de balanço e CDA, de maneira similar ao que foi observado para o zinco. A dieta à base de soja, entretanto, mostrou balanço e CDA positivos para cobre e zinco. Por outro lado, somente a dieta Controle teve resposta positiva no teste de balanço de zinco, apesar da taxa de R/A para as dietas ¿Ouro Branco¿ e ¿Pérola¿ ter sido alta. Juntando estes dados, os índices sugerem que a biodisponibilidade de cálcio e magnésio não foi afetada pelos fitatos presentes nestas fontes alimentares, porém, para o zinco e o cobre este efeito foi evidente. As medidas histomorfométricasm, entretanto, não detectaram nenhuma diferença entre os diversos tratamentos à base de feijão e soja. Além disso, pode ser concluído que a baixa ingestão de cálcio dos grupos alimentados com leguminosas, acarretou alterações no sistema esquelético, tais como alta redução do platô da cartilagem de crescimento e na espessura do osso em si, mesmo que a retenção de cálcio no fêmur dos animais tratados com as dietas ¿Pérola¿ e soja tenham sido significativa / Abstract: Given the importance that dry beans have in the traditional Brazilian diet, the Brazilian Agency for Agricultural Research (EMBRAPA) has developed new cultivars of common beans (P. vulgaris) by means of classical breeding expecting to meet higher agronomic and culinary standards for the domestic market. Nevertheless, the nutritional characteristics of these new cultivars are incompletely documented, both in terms of macro/micro nutrient contents and biological value. The objective of the present work was to evaluate the nutritive value of seven new dry bean cultivars: Jalo Precoce, Radiante, Vereda, Pérola, Timbó, Valente e Ouro, from the Embrapa Rice and Bean Research Center in Goiás. After routine proximal composition, their amino acid profiles and key mineral components (calcium, iron, magnesium, zinc and copper) were determined. The results show that substantial and significant (P<0,05) variations occur in protein and carbohydrates among all the cultivars. Protein contents varied between 21 and 28%; Radiante, Jalo Precoce and Ouro Branco having the highest contents. In turn, the total carbohydrate contents ranged between 55,4 for Radiante, to 62.1%, for Valente, whereas the lipids remained rather constant, varying from 2.4 to 2.8%. The proteins of all these new cultivars met the international standard requirements for histidine, valine, isoleucine, leucine and phenylalanine plus tyrosine. Additionally, the cultivars Valente and Ouro Branco stood out because of their contents for lysine and threonine that also met the requirements. As in the case of all leguminous seeds, all the cultivars were low in the sulfurcontaining amino acids, methionine and cysteine. No statistical differences were noticed among the cultivars in terms of Mg, Fe and Zn, but the concentrations of Ca and Cu did vary significantly (P<0.0001). Iron contents varied from 4.7 in Ouro Branco, to 5.5mg/100g in Vereda and Valente, while copper ranged from 0.5 in Ouro Branco, to 0.9mg/100g, in Radiante, and the zinc contents went from 2.4 (Ouro Branco and Pérola) to 3.1mg/100g (Jalo Precoce). Insofar as calcium is concerned, Pérola (486), Vereda (428) and Valente (387mg/100g) were the richest, whereas Jalo Precoce, Pérola and Valente stood out because of their high contents of magnesium (129 ¿ 117mg/100g). Considering the fact that all the cultivars were grown under strictly controlled conditions of soil, weather and agricultural practices, these results can be taken as highly representative of the individual compositional characteristics and could be concluded that among the new cultivars, the Jalo Precoce, Ouro Branco and Valente are the most advantageous in terms of nutrient composition. Then, three of those new Brazilian dry bean (P. vulgaris) cultivars were selected because of their nutrient content for mineral bioavailability evaluation, and compared with the classical soybean (Glycine max) cultivar Conquista. The three cultivars were the white bean ¿Ouro Branco¿, a black bean ¿Valente¿ and the Carioca type ¿Pérola¿ and the minerals tested were the least commonly studied: calcium, magnesium, copper and zinc. The metabolic balance, apparent digestibility coefficient (ADC) and the ratio of retention to absorption (R/A) for these minerals were the parameters determined using young Wistar rats (21d). The animals received for twelve days diets consisting exclusively of the cooked beans or the roasted soybean flour. The phytate levels of the diets were determined by the colorimetric method of Latta & Eskin and the minerals by atomic absorption in the diets, feces, urine and femur. Additionally and in order to evaluate the impact of the diets on the cortical bone growth of the tibias, histomorphometric measurements of the height and thickness of the cartilage were made. The results showed that the legume-based diets statistically differed from each other in terms of calcium (275 ¿ 480 mg/100g), magnesium (108 ¿ 262 mg/100g), copper (0.50 ¿ 1.06 mg/100g) and zinc (2.4 ¿ 5.0mg/100g), and that the diets made up of the Carioca ¿Pérola¿ and the Black ¿Valente¿, as well as soy bean were those with the highest mineral contents. The dry bean diets exhibited low concentrations of phytates (3.1 ¿ 3.9mg/g), especially with respect to the soybean (6.5mg/g). Statistical analysis of the mineral balance detected significant differences between the biological indices obtained by the dry-bean and the soybean treatments. The dry bean diets ¿Valente¿ and ¿Pérola¿ exhibited higher balance for calcium (22 ¿ 25mg/d), in comparison to ¿Ouro Branco¿ (12.8mg/d) and the soybean (14.4mg/d) diets. The ratio of retention to absorption (R/A), however, was greatest for the soybean (99.0%), followed by the ¿Valente¿ (96.7%) diet. With regard to the ADC for calcium, in turn, the ¿Pérola¿ ranked first (92.5%) among all treatments and the Control. The evaluation indices revealed that the soybean had the highest magnesium balance (4.7mg/d) among all the dry bean diets, whereas the ADC was best for the ¿Pérola¿ (82%) diet, although the R/A ratios were better for the ¿Valente¿ (7.1%) and soybean (6.8%), while the ¿Ouro Branco¿ and Control gave negative indices (balance and R/A values). With regard to copper, all the dry bean diets gave negative balance and ADC indices, something similar to what was observed for zinc. The soy bean diet, however, showed positive balance and ADCs for both copper and zinc. On the other hand, only the Control diet gave a positive response in the balance test for zinc, although the R/A ratios of the ¿Ouro Branco¿ and ¿Pérola¿ diets were the highest. Taken together, these indices suggest that the little studied calcium and magnesium bioavailability from dry beans is not affected by the phytates present in this staple food, but a suppressive effect on zinc and copper was clear. Histomorphometric measurements, however, did not detect any differences between the various dry bean treatments or the soybean. In addition, it could be concluded that the low calcium intake by dry bean-fed groups had profound consequences to the skeleton system of the animals, such as the height reduction of the cartilaginous grow plates and the thickness of the bone itself, in spite of the femur calcium retention having been significant in the ¿Pérola¿ and soybean diets / Doutorado / Nutrição Básica Experimental e Aplicada à Tecnologia de Alimentos / Doutor em Alimentos e Nutrição

Made available in DSpace on 2014-07-29T16:16:36Z (GMT). No. of bitstreams: 1 dissertacao aline silva.pdf: 641990 bytes, checksum: f01c5af83294c71fd63825142856d782 (MD5) Previous issue date: 2009-09-14 / Many species from Cerrado remain unknown in their biochemical characteristics and in their potential for food processing. This work had as objective to characterize the almond of chichá from the specie Sterculia striata A. St. Hill & Naudin, originated from Corrente (Piauí) in their biochemical components, analyze the changes in the chemical composition and the amounts of antinutritional factors after roasting. The almond of chicha presented high levels of protein, fiber, phosphorus, copper and manganese. The roasting held to 205°C for 11 minutes reduced the content of humidity from 6.0% to 2.1% and phytates from 10,6mg/g to 5.5 mg/g. There were no lectins, tannins, trypsin inhibitors and alpha-amylase was also a important result obtained. It was verified the absence of the enzymes polyphenol oxidase and peroxidase and a content of phenolics in the range of 107.7 mg/100g and 108.9 mg/100g in the raw and roasted almond respectively. The almond of chicha presented polyunsaturated fatty acids linoleic (3,8%) and linolenic acid (2,3%), high levels of monounsaturated (40%) and saturated (34.5%) and presence of fatty acids cyclopropenoid. Therefore, the almond chichá is rich in nutrients; however it is necessary to investigate the effects of eating cyclopropenoid in the concentration found in these almonds. This work reinforces the necessity of conservation and in deep studies of native plants from Cerrado / Muitas espécies do Cerrado ainda são pouco conhecidas, assim como suas características bioquímicas e seu potencial para processamento de alimentos. Este trabalho teve como objetivo caracterizar bioquimicamente a amêndoa de chichá da espécie Sterculia striata A. St. Hill & Naudin, oriunda de Corrente (Piauí) e verificar alterações na composição química e no teor de fatores antinutricionais após torrefação. A amêndoa de chichá mostrou-se ser um alimento com alto teor de proteínas, fibras, fósforo, cobre, zinco e manganês. A torrefação realizada a 205°C por 11 min resultou em perda de umidade de 6,0% para 2,1% e diminuição do fitato de 10,6 mg/g para 5,5 mg/g. A ausência de fatores antinutricionais como lectinas, taninos e inibidores de tripsina e alfa-amilase foi também um resultado importante. Verificou-se a ausência de peroxidase e polifenoloxidase e conteúdo de compostos fenólicos de 107,7 mg/100g e 108,9 mg/100g para amêndoas cruas e torradas, respectivamente. Por fim, têm-se ácidos graxos poliinsaturados linoléico (3,8%) e linolênico (2,3%), altos teores de monoinsaturados (40%) e saturados (34,5%) e presença de ácidos graxos ciclopropenoídicos. Assim, pode-se afirmar que a amêndoa de chichá é um alimento rico em nutrientes, mas é preciso investigar os efeitos da ingestão de ácidos ciclopropenoídicos na concentração encontrada. Este trabalho reforça a necessidade de conhecer e conservar as plantas nativas do Cerrado