Le manioc (Manihot esculenta Crantz) est une plante, dont les tubercules servent de nourriture de base pour près de 800 millions de personnes sous les tropiques. Malgré sont importance, ce tubercule présente deux inconvénients majeurs qui limitent leur utilisation en alimentation humaine : Une toxicité liée à la présence de composés cyanés, et une faible teneur en protéines. Dans l'optique de lever ces contraintes, plusieurs procédés de transformation ont étés mis au point par l'expérience des populations et des améliorations ont été apportées par la recherche. La plupart de ces procédés incluent une étape critique de fermentation spontanée. Cette étape présente quelques problèmes dus à sa dépendance vis à vis des microorganismes épiphytes. Ces problèmes sont notamment la présence de quantités résiduelles de composés cyanés non négligeables (100 à 170ppm) un temps de ramollissement très long (4 à 5 jours) et des produits dérivés de qualités nutritionnelles et hygiéniques douteuses. Dans l'optique d'améliorer ces procédés traditionnels de rouissage du manioc cyanogène, une centaine de microorganismes ont été isolés des cossettes de manioc séché, sur la base de leurs potentialités à produire la β-glucosidase, l'α-amylase, la pectine hydrolase et la polygalacturonase. Ces enzymes sont les plus impliqués dans les phénomènes biochimiques et physico-chimiques ayant lieu au cours du rouissage du manioc. Ils sont responsables de la dégradation des composés cyanés et du ramollissement des tubercules par dégradation des pectines qui constituent le squelette du tubercule. 60% des microorganismes isolés et identifiés ont été productrices d'α-amylases, 46% de β-glucosidase, et 36%de pectinases. Les capacités des différentes souches sélectionnées à produire ces principaux enzymes, ont ensuite été évaluées. Parmi ces microorganismes, deux se sont avérés particulièrement intéressantes : Une souche de bactérie lactique identifiée comme étant Lactobacillus plantarum avec une activité α-amylase de l'ordre de 17000±100UE/ml, et une activité ß-glucosidase de 2000±300µmol CN-/mn; Une moisissure identifiée comme étant Rhizopus oryzae à activité α-amylase de 20000±500UE/ml et activité pectine hydrolase de 2,3±0,2 µmol COO-/mn/mg. Ces deux souches ont été sélectionnées et utilisées comme ferments en culture mixte avec une biomasse initiale de 106ufc/g manioc frais de Lactobacillus plantarum et 103spores/g manioc frais de Rhizopus oryzae pour la fermentation des tubercules de manioc. Les résultats obtenus ont indiqué une acidification rapide avec un pH avoisinant 3,5±1 après 24 heures de rouissage, un ramollissement plus rapide (12±3mm/5s après 48 heures de rouissage). Mais le rouissage naturel s'est avéré meilleur du point de vue dégradation des composés cyanés. En effet la très forte acidification obtenue avec le ferment dès la 24eme heure, a provoqué un blocage du processus de dégradation des glucosides cyanogénétiques au niveau de la cyanohydrine, composé par ailleurs aussi toxique que l'acide cyanhydrique. Une étude de l'influence de quelques paramètres externes nous ont permis de noter que la température optimale de croissance de Lactobacillus plantarum était de 40±5°C pour un pH optimal de 6 alors que Rhizopus oryzae avait une température optimale de croissance à 30±7°C et un pH optimal de 5. Les essais de production de biomasses, indiquent que le meilleur milieu de multiplication des germes s'est avéré être le milieu CTM de Law et al., avec pour substrat de la farine de son de manioc mélangée au milieu nutritif de Meyer.<br />Dans l'optique de mettre à la disposition des populations ce ferment, des essais de conservation des souches par séchage ont indiqué que le fluide de protection indiqué pour avoir des taux de survies acceptables au terme du séchage était le glycérol, alors que le support le plus adapté a été l'amidon de manioc. On a ainsi obtenu plus de 75±5% de survivants après séchage sur lit convectif à une température de 35°C sous un débit d'air de 10 m3/h pour tous les deux germes. Ce ferment composé de Lactobacillus plantarum (106ufc/g) et Rhizopus oryzae (103spores/g), sur support amidon a induit contrairement aux microorganismes frais, une diminution effective du taux de glucosides totaux résiduels (5% en fin de rouissage) diminution corollaire à l'ajout du NaOH réalisé après la 24ème heure, afin de contourner l'effet néfaste du pH acide sur la dégradation de la cyanohydrine.

The effects of peptides derived from β-lactoglobulin and α-lactalbumin on metabolic syndrome were studied. α-lactalbumin and β-lactoglobulin were hydrolyzed with trypsin, alcalase, flavourzyme, or a combination of alcalase and flavourzyme and fractionated. Angiotensin coverting enzyme inhibition of the < 1 kDa fraction of alcalase hydrolyzed β-lactoglobulin was 95 %. Antioxidant activity of the < 1 kDa fraction of β-lactoglobulin hydrolyzed with a combination of alcalase and flavourzyme was 18 %. Stimulated adipocytes incubated with the < 1 kDa fraction of β-lactoglobulin hydrolyzed with either trypsin or alcalase produced 30 pg/mL of interleukin 6. Adiponectin and glucose transporter type 4 secretions increased 1.1 and 0.86 fold respectively during incubation with the < 1 kDa fraction of β-lactoglobulin hydrolyzed with a combination of alcalase and flavourzyme. Results indicate that β-lactoglobulin peptides formed with alcalase and a combination of alcalase and flavourzyme influence markers associated with metabolic syndrome and may be useful as functional foods or nutraceuticals.

The effects of peptides derived from β-lactoglobulin and α-lactalbumin on metabolic syndrome were studied. α-lactalbumin and β-lactoglobulin were hydrolyzed with trypsin, alcalase, flavourzyme, or a combination of alcalase and flavourzyme and fractionated. Angiotensin coverting enzyme inhibition of the < 1 kDa fraction of alcalase hydrolyzed β-lactoglobulin was 95 %. Antioxidant activity of the < 1 kDa fraction of β-lactoglobulin hydrolyzed with a combination of alcalase and flavourzyme was 18 %. Stimulated adipocytes incubated with the < 1 kDa fraction of β-lactoglobulin hydrolyzed with either trypsin or alcalase produced 30 pg/mL of interleukin 6. Adiponectin and glucose transporter type 4 secretions increased 1.1 and 0.86 fold respectively during incubation with the < 1 kDa fraction of β-lactoglobulin hydrolyzed with a combination of alcalase and flavourzyme. Results indicate that β-lactoglobulin peptides formed with alcalase and a combination of alcalase and flavourzyme influence markers associated with metabolic syndrome and may be useful as functional foods or nutraceuticals.

Submitted by MARCOS LEANDRO TEIXEIRA DE OLIVEIRA (marcosteixeira@ufv.br) on 2018-08-15T18:29:36Z No. of bitstreams: 1 texto completo.pdf: 1510730 bytes, checksum: 754986579a43c0d4c754880228c9a25d (MD5) / Made available in DSpace on 2018-08-15T18:29:36Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1510730 bytes, checksum: 754986579a43c0d4c754880228c9a25d (MD5) Previous issue date: 2016-08-25 / A brusone, causada por Pyricularia oryzae, é considerada uma doença economicamente importante para trigo na América do Sul. Uma das estratégias de manejo para minimizar as perdas causadas por essa doença é o uso de cultivares resistentes. Alternativamente, pode-se utilizar indutores de resistência para aumentar o nível basal de resistência do trigo à brusone. O presente estudo teve como objetivos: i) determinar as alterações fisiológicas e bioquímicas em plantas de trigo pulverizadas com uma concentração não fitotóxica do ácido α-picolinico (AP), o qual é uma toxina não seletiva produzida por P. oryzae e ii) verificar a manipulação metabólica exercido por P. oryzae quando infectando cultivares de trigo com diferentes níveis de resistência basal à brusone. Nas folhas de trigo pulverizadas com uma concentração não fitotóxica de AP (0.1 mg mL -1 ), os sintomas da brusone desenvolveram menos em associação com um melhor desempenho fotossintético, melhoria do metabolismo antioxidante e redução nas concentrações de H 2 O 2 , O 2 ●- e MDA. As cultivares BR-18 e EMBRAPA- 16 foram mais resistentes à brusone do que a cultivar BRS-Guamirim. O desempenho fotossintético das plantas infectadas foi alterado devido a limitações de natureza difusiva e bioquímica para uma eficiente fixação do CO 2 . Durante a fase assintomática da infecção por P. oryzae, mudanças drásticas no metabolismo de carboidratos e nos níveis de aminoácidos, compostos intermediários do ciclo de Krebs e poliaminas ocorreram nas plantas das três cultivares sugerindo, assim, uma manipulação metabólica exercida por P. oryzae. No entanto, um metabolismo antioxidativo mais eficiente foi importante para neutralizar os efeitos deletérios da infecção por P. oryzae em associação com maiores atividades da fenilalanina amônia liase e polifenoloxidase e maiores concentrações de compostos fenólicos e lignina. Com base nesses resultados e possível concluir que a concentração não fitotóxica de AP foi capaz de potencializar a defesa das plantas de trigo e reduzir a severidade da brusone. A infecção do trigo por P. oryzae ocasionou distúrbios no metabolismo primário das plantas e alguns deles foram semelhantes entre as cultivares independentemente do nível basal de resistência delas. / Blast, caused by Pyricularia oryzae, has become an economically important disease in wheat in South America. One of the management strategies for minimizing the losses caused by blast includes the use of resistant cultivars. Alternatively, the use of inducers of resistance showed the potentiation to increase wheat resistance to blast. This study aimed: i) to determine the physiological and biochemical alterations in wheat plants sprayed with a non-phytotoxic concentration of α-picolinic acid (PA), which is a non-host selective toxin produced by P. oryzae and ii) to establish the degree of metabolic manipulation exerted during the infection by P. oryzae on plants from cultivars with different levels of basal resistance to blast. The spray of leaves of plants with a non-phytotoxic concentration of PA (0.1 mg mL -1 ) resulted in less blast symptoms in association with a better photosynthetic performance, an improvement on the antioxidant metabolism and reduced concentrations of H 2 O 2 , O 2 ●- and malondialdehyde. The cultivars BR-18 and EMBRAPA-16 were more resistant to blast in comparison to cultivar BRS-Guamirim. The photosynthetic performance of the infected plants was altered due to diffusional and biochemical limitations for CO 2 fixation. During the asymptomatic phase of P. oryzae infection, drastic changes in the carbohydrates metabolism and on the levels of amino acids, intermediates compounds of Krebs cycle and polyamines occurred on plants from the three cultivars suggesting a metabolic manipulation exerted by the pathogen. However, amore efficient antioxidant metabolism was able to help the wheat plants to counteract against the deleterious effects of P. oryzae infection in association with great phenylalanine ammonia lyases and polyphenoloxidases activities and high concentrations of phenolics and lignin. Based on this information, it is possible to conclude that a non- phytotoxic concentration of PA elicited the activation of host defense mechanisms that reduced blast severity. Likewise, the infection of leaves by P. oryzae induced remarkable disturbances in the primary metabolism and some of them were conserved among the cultivars regardless of their basal level of resistance to blast.

The blast rice, caused of fungus Pyricularia oryzae is the most important rice crop disease and can cause up to 100% crop damage. The cultural, varietal and chemical management at the field level and the monitoring and knowledge of in vitro blast isolates sensitivity are essential tools for better understanding and control of this disease. The objective of this project was to evaluate the effect of the fungicides most used in blast control in three cultivars of irrigated rice in two sowing seasons and to determine in vitro the sensitivity of the mycelial growth of two isolates of P. oryzae to the same fungicides: Triciclazole, Tebuconazole, Azoxystrobin, Trifloxystrobin + Tebuconazole and Casugamycin. For that, an experiment was carried at the field level in the 2015/2016 cropseason in Santa Maria-RS and another experiment in the laboratory. In the first experiment the cultivars used were Guri INTA CL, Puitá INTA CL and IRGA 424 RI sown in the beginning and end of the recommended period and submitted to the same fungicides. The second experiment tested the fungicides at concentrations of 0, 0.01, 0.1, 1, 10 and 100 ppm in two isolates of blast. It was concluded that the cultivar IRGA 424 IR showed to be resistant to blast and that for the maintenance of productive levels in the cultivars susceptible to P. oryzae, especially in the second sowing season, the adoption of more efficient fungicides is indispensable. In relation to mycelial growth in vitro, the first insensitivity of a P. oryzae isolate to the active ingredients Triciclazol and Azoxystrobin was verified at the national level. / A brusone, causada pelo fungo Pyricularia oryzae, é a doença mais importante da cultura do arroz, podendo causar até 100% de prejuízos à cultura. O manejo cultural, varietal e químico a nível de campo bem como o monitoramento e conhecimento da sensibilidade in vitro de isolados de brusone, são ferramentas imprescindíveis para o melhor entendimento e controle desta doença. O objetivo deste projeto, foi avaliar o efeito dos fungicidas mais utilizados no controle de brusone, em três cultivares de arroz irrigado, em duas épocas de semeadura e determinar in vitro a sensibilidade do crescimento micelial de dois isolados de P. oryzae aos mesmos fungicidas: Triciclazol, Tebuconazole, Azoxistrobina, Trifloxistrobina + Tebuconazole e Casugamicina. Para tal, foi realizado um experimento em nível de campo na safra 2015/2016 em Santa Maria-RS e outro experimento em laboratório. No primeiro experimento foram utilizadas as cultivares GURI INTA CL, PUITÁ INTA CL e IRGA 424 RI, semeadas no início e no final da época recomendada e essas submetidas aos mesmos fungicidas. O segundo experimento, testou os fungicidas anteriormente citados nas concentrações de 0, 0.01, 0.1, 1, 10 e 100 ppm em dois isolados de brusone. Concluiu-se que a cultivar IRGA 424 RI mostrou-se resistente à brusone e que para a manutenção dos patamares produtivos nas cultivares suscetíveis a P. oryzae, principalmente na segunda época de semeadura, torna-se necessário a adoção de fungicidas mais eficientes. Em relação ao crescimento micelial in vitro, constatou-se à nível nacional, a primeira insensibilidade de um isolado de P. oryzae aos ingredientes ativos Triciclazol e Azoxistrobina.

Vegetable oil is more difficult to degrade by microorganisms in comparison to carbohydrates and protein. Thus, it creates serious environmental and health concerns if oil-derived waste streams produced by restaurants and industries remain untreated. In this study, a strategy has been developed to grow filamentous fungi in pure olive oil so that it can be used as a bench-mark for growth in olive oil mill sidestreams. The growth of different strains (Aspergillus oryzae, Neurospora intermedia and Rhizopus oryzae) was tested in pure olive oil. A pre-germination stage using glucose as carbon and energy source, or the addition of yeast extract, were found necessary for successful fungal growth in olive oil. Here, A. oryzae showed a superior performance in comparison to N. intermedia and R. oryzae. Medium pH did not impact A. oryzae growth in olive oil, whereas a concentration higher than 40 g/L of the latter impaired the growth of the ascomycete. Obtained biomasses from A. oryzae and N. intermedia cultivations in olive oil were analyzed and compared for protein, fat, ash, and alkali-insoluble material (cell wall content), where the presence of olive oil had a steering effect. The fungal biomass of A. oryzae, obtained from cultivation in the absence of olive oil, contained 0.33% fat and 48% protein, whereas the respective values in the presence of olive oil were 31% and 14%. Similar trends on fat and protein contents were observed for the biomass of N. intermedia. Sudan black staining was also performed on fresh biomass which clearly indicated the presence of oil globules inside the fungal cells. This research can be a fundamental step towards treatment of oil-based waste streams, which entails high-energy and costs if treated, or environmental impacts during informal discharges. Moreover, the fact that the composition of fungal biomass can be steered through addition of olive oil increases the versatility of the originated biomass for various applications, namely in feed, food and biofuel production.

in English β-N-acetylhexosaminidase (EC 3.2.1.52) belongs to exoglycosidase, and is one of the most abundant enzymes found in organisms from bacteria to human. The fungal β-N-acetylhexosaminidase from Aspergillus oryzae is composed of propeptide and catalytic domain. The propeptide is noncovalently associated with the catalytic domain of the enzyme. Propeptide is essential for the enzyme activity. While the structure of the catalytic domain was desidned by homology modeling, the structure of the propeptide has not been resolved yet. In this study, the position where the propeptide is associated with the catalytic domain, was uncovered. Presented work consists of two parts. First part deals with optimization of production conditions, purification and crystallization of β-N-acetylhexosaminidase from the filamentous fungi Aspergillus oryzae. Second part is devoted to the study of interaction between propeptide and catalytic domain, which was characterized by chemical cross-linking and high-resolution mass spectrometry. It was found that the structural changes of the catalytic domain depend on the presence of the propeptide molecule. Moreover the region of propeptide-catalytic domain interaction was revealed.

Twenty-four early to midlactation Holstein cows were allocated to one of three treatments. Treatments consisted of: 1) basal ration plus 60 g wheat bran per day per head (control); 2) basal ration plus 57 g yeast culture per day per head; 3) basal ration plus 57 g yeast culture plus 3 g Aspergillus oryzae per day per head. Feed intake and milk yield were recoreded daily and milk composition and body weights were recorded weekly. Feed and fecal samples were recorded weekly. Feed and fecal samples were collected to determine apparent nutrient digestibility. No statistical difference was observed in milk yield among treatments. Percent fat, protein, and solid non fat of milk samples for the control fed group was significantly higher than the other treatment groups. Apparent crude protein, acid detergent fiber, and neutral detergent fiber digestibilities for cows fed the fungal culture treatments were significantly higher. Six rumen-fistulated Holstein cows were randomly assigned to one of three treatments in a 3x3 replicated Latin square design. Rumen pH was significantly lower for cows fed treatment two. Total volatile fatty acid and ammonia nitrogen concentration were higher for cows fed treatment two. Three rumen-fistulated Holstein cows were assigned one of three treatments in a 3x3 Latin square design for an in situ study. Approximately 15 g of grass hay, low quality alfalfa, and high quality alfalfa were sealed in nylon bags and put into the rumen. Samples were analyzed for dry matter, acid detergent fiber, neutral detergent fiber, and crude protein. Regardless of substrate used, dry matter disappearance was lower for control. Regardless of treatment ration fed, high quality alfalfa as a substrate was significantly higher in dry matter disappearance.

No description available.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-04-26T13:48:20Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_cassyo_de_araujo_rufino.pdf: 1782663 bytes, checksum: 48211778344c5f757ff01c4c9282d9b8 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-05-02T18:29:29Z (GMT) No. of bitstreams: 2 tese_cassyo_de_araujo_rufino.pdf: 1782663 bytes, checksum: 48211778344c5f757ff01c4c9282d9b8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-05-02T18:31:26Z (GMT) No. of bitstreams: 2 tese_cassyo_de_araujo_rufino.pdf: 1782663 bytes, checksum: 48211778344c5f757ff01c4c9282d9b8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-05-02T18:31:51Z (GMT). No. of bitstreams: 2 tese_cassyo_de_araujo_rufino.pdf: 1782663 bytes, checksum: 48211778344c5f757ff01c4c9282d9b8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-03-05 / Sem bolsa / A utilização de produtos naturais conjuntamente com nanopartículas de prata tem atraído interesse de pesquisadores, tanto do ponto de vista tecnológico quanto cientifico devido à ação desses produtos no controle de doenças e pragas. O objetivo desse trabalho foi testar produtos naturais (associados ou não a nanoprata) com ação antimicrobiana sobre Bipolaris oryzae, in vitro, e no tratamento de sementes de arroz. Para o estudo dos compostos contendo quitina e própolis adicionadas opcionalmente nanoprata foram utilizados sementes de arroz na cultivar IRGA 424 inoculadas com um isolado de Bipolaris oryzae. Os compostos utilizados são naturais, têm propriedades antiadesivas e atividade antimicrobiana contra microrganismos, possuem propriedades biocompatíveis e não fitotóxicas aos vegetais em geral. O processo de sínteses dos compostos contendo própolis, quitina e nanoprata, foi realizado por meio da aplicação de sonicação para a dispersão e mistura entre as substâncias. Foram testados os produtos no desenvolvimento e crescimento do fungo Bipolaris oryzae em placas de petri e em sementes de arroz infectadas. Os tratamentos foram: quitina, própolis, prata, quitina+própolis, quitina+própolis+nanoprata, quitina+prata, própolis+nanoprata e testemunha (T - sementes sem tratamentos), totalizando neste estudo 10 tratamentos. Logo a eficiência dos produtos e suas soluções foram testados por meio do blotter test (teste de sanidade de sementes), porcentagem de inibição e diâmetro de crescimento do fungo, germinação e comprimento de plântulas das sementes tratadas com os produtos. Os resultados demonstram que os oligômeros de quitosana e os compósitos de quitosana com extrato de própolis e nanopartículas de prata controlam o crescimento do patógeno. A importância deste achado reside no fato de que é um passo em direção ao objetivo de diminuir o uso de fungicidas químicos. A partir de testes de germinação poderá concluir-se que os melhores resultados foi o tratamento com quitosano/prata compósito NPs, com uma taxa de germinação de 91%. Os novos materiais poliméricos são considerados como muito promissores para o controle de Bipolaris oryzae no arroz, tornando-se assim alternativas ecológicas aos fungicidas químicos. / The use of natural products in conjunction with silver nanoparticles has attracted interest of many researches, from both scientific and technological point of view due to their action for disease and pests control. The objective was to test natural products (with or without nanosilver) with antimicrobial action against Bipolaris oryzae, in vitro, and in the treatment of rice seeds. To study the compounds containing chitin and propolis optionally added nanosilver rice seeds were used in IRGA inoculated with an isolated 424 Bipolaris oryzae. The compounds used are natural, have anti-adhesive properties and antimicrobial activity against microorganisms, have biocompatible properties and not phytotoxic to plants in general. The process of synthesis of compounds containing propolis, chitin and nanosilver was performed by applying the sonication for the dispersion and mixing of the substances. Products were tested on the development and growth of Bipolaris oryzae fungus in petri dishes and infected rice seeds. The treatments were: chitin, propolis, silver, chitin + propolis, chitin + Propolis + nanosilver, chitin + silver, propolis + nanosilver and control (T - treatments seeded), totaling 10 treatments in this study. The results were conclusive in showing that chitosan oligomers and the composites of chitosan with propolis extract and silver nanoparticles led to a remarkable reduction in growth of the pathogen. The importance of this finding lies in the fact that it is a step towards the goal of decreasing the use of chemical fungicides in plant pathology. From germination tests it could be concluded that the best results was the treatment with chitosan/silver NPs composite, with a germination rate of 91%. The novel polymeric materials can thus be deemed as very promising for the control of the Bipolaris oryzae of rice and pave the way towards eco-friendly alternatives to chemical fungicides.