Diversos estudos têm mostrado que alimentos adicionados de probióticos e prebióticos, em quantidades suficientes, trazem benefícios à saúde humana. Estes benefícios se devem, fundamentalmente, ao seu efeito na manutenção da microbiota intestinal benéfica. Paralelamente, ocorre aumento do consumo de derivados do açaí em grandes centros urbanos no Brasil e até mesmo no exterior, principalmente a sua polpa e derivados, que se deve, principalmente, ao alto valor calórico da fruta e à presença de pigmentos antioxidantes. O presente trabalho teve como principal objetivo desenvolver um mix de açaí que apresente aspectos nutricionais e sensoriais semelhantes ao produto tradicional, porém com propriedade funcional suplementar, por se tratar de um alimento probiótico, prebiótico ou simbiótico. Quatro tratamentos foram produzidos (três vezes cada um deles), todos contendo mix de açaí congelado: M1 (controle), M2 (com L. acidophilus La-5 + B. animalis subsp. lactis Bb-12), M3 (com inulina) e M4 (com L. acidophilus La-5 + B. animalis subsp. lactis Bb-12 + inulina) e armazenados a -18 °C por até 3 meses. Após 1, 7, 14, 21, 28, 56 e 84 dias de armazenamento, foram determinadas as populações de microrganismos probióticos, o pH e a cor instrumental dos produtos. O teor de umidade foi determinado após a fabricação dos produtos (dia 0). Adicionalmente, após 7, 42 e 84 dias de armazenamento, foi realizada análise sensorial (aceitabilidade). Foi determinada a composição centesimal dos produtos, a partir de amostras mantidas liofilizadas. Todos os resultados foram comparados, através de análise estatística. A diferenciação na composição centesimal e no teor de sólidos entre as formulações decorreu da adição da fibra prebiótica. Com relação à evolução da cor e do pH, não houve variação importante ao longo do tempo, assim como entre os diferentes mixes de açaí estudados. Os produtos obtiveram boa aceitabilidade sensorial por parte dos provadores (médias das notas acima de 7, na maioria das formulações). Houve maior aceitabilidade dos mixes suplementados com o ingrediente prebiótico e o mix simbiótico recebeu notas médias significativamente mais elevadas (p < 0,05) em dois períodos de armazenamento (7 e 84 dias), quando comparado aos demais. O tempo de armazenamento não influenciou a avaliação sensorial significativamente (p > 0,05). Durante o período de armazenamento proposto, os probióticos apresentaram sobrevivência satisfatória (populações superiores a 108 UFC por porção diária), sob o ponto de vista da legislação brasileira vigente, com exceção do B. animalis subsp. lactis no mix não suplementado de inulina. O mix de açaí apresentou-se como boa matriz para veicular os microrganismos probióticos testados e a inulina contribuiu para uma maior aceitabilidade sensorial do produto. / Some studies have been showing that food products supplemented with probiotics and prebiotics, in sufficient amounts, result in benefits to human health. These benefits are related, fundamentally, to their effect on the healthy intestinal microbiota maintenance. Concomitantly, consumption of açai food products, mainly its pulp and derivatives, is increasing in urban centers in Brazil and also abroad. The high caloric value and the presence of antioxidant pigments are responsible for its higher consumption. The main aim of the present study was to develop an açai mix with nutritional and sensory aspects similar to the conventional product, but with a supplementary functional property, by being a probiotic, prebiotic or synbiotic food. Four trials where produced (three times each), all containing frozen açaimix: M1 (control), M2 (supplemented with L. acidophilus La-5 + B. animalis subsp. lactis Bb-12), M3 (supplemented with inulin) and M4 (supplemented with L. acidophilus La-5 + B. animalis subsp. lactis Bb-12 + inulin) and stored at -18 °C for up to 3 months. After 1, 7, 14, 21, 28, 56 and 84 days of storage, the products were analyzed for probiotic microorganisms populations, pH and instrumental color. The moisture content was determined soon after production (on day 0). Additionally, after 7, 42 and 84 days of production, sensory evaluation (acceptability test) was carried out. Finally, compositional analyses proceeded from freeze-dried samples. Results were compared through statistical analysis. Differences in compositional analyses and moisture content between trials were attributed to the presence or absence of inulin. Instrumental color and pH evolution were not influenced by the storage period and also no differences between the different acaimixes studied were observed (p > 0,05). Products were well accepted by the panellists, and mean scores were often above 7. Prebiotic fiber inulin improved overall acceptability of açaimixes and the synbiotic mix obtained higher mean score (p < 0.05) in two storage periods (7 and 84 days), when compared to the others. The storage period did not influence sensory evaluation significantly (p > 0.05). During the storage period studied, probiotics showed a satisfactory survival (viability higher than 108 cfu for the products daily portion), according to the current Brazilian legislation, except for B. animalis subsp. lactis in the mix not supplemented with inulin. The açaimix showed to be a good matrix for the probiotic microorganisms tested and inulin contributed for a higher acceptability of the product.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / The objective of this study was to develop a probiotic cashew apple juice ready to drink and in the dehydrated form through spray drying. The first stage of the study was the optimization of Lactobacillus casei NRRL B-442 cultivation in cashew apple juice, to optimize the proper inoculum amount and the fermentation time. The optimum conditions for probiotic cashew apple juice production were: initial pH 6.4, fermentation temperature of 30ÂC, inoculation level of 7.48 log CFU/mL (L. casei) and 16 h of fermentation process. Cashew apple juice showed to be as efficient as dairy products for L. casei growth. In a second stage, the stability of probiotic cashew apple juice stored for 42 days at 4ÂC was evaluated. Analyses were conducted in the non fermented cashew apple juice (control), and in the fermented juices with L. casei NRRL B-442, with 8% (w/v) of sucrose (sugar table), after fermentation, and without the addition of sugar. The viability of the probiotic bacteria, sugars and organic acids content, color, antioxidant and enzymatic activity, and sensory characteristics were evaluated during the storage. Viable cell counts increased in the probiotic cashew apple containing sucrose along the storage period. Moreover, the fermentation lead to the preservation of the ascorbic acid content, which had a less intense reduction in the fermented cashew apple juices compared to the non fermented sample. The antioxidant activity and total polyphenolic compounds of cashew apple juice had a similar trend. Browning reactions and nutritional breakdown caused by enzymes were minimized in the fermented samples during storage. In these samples, a higher reduction of the enzymatic activity of polyphenoloxidase and peroxidase activity was observed. During the storage, the increase in the chroma values indicated that yellowness was reinforced, being well accepted by consumers. The sensory attributes (aroma, flavor, acidity and color) of probiotic cashew apple juice were positively influenced by storage under refrigeration for 42 days. In the third stage of the research, the effects of dehydration by spray drying in cashew apple juice containing L. casei NRRL B-442 was assessed and the influence of storage temperature on the viability of L. casei NRRL B-442 and physical properties of the powder were evaluated during 35 days of storage. The drying agents used were: 20% (w/v) maltodextrin or 10% (w/v) maltodextrin + 10% (w/v) arabic gum. The powder of probiotic cashew apple juice showed satisfactory levels of L. casei survival, during drying. During storage, the addition of 10% (w/v) maltodextrin + 10% (w/v) arabic gum kept microbial viability within satisfactory levels when the powder was subjected to cooling at 4ÂC. However, greater differences in the reconstituted powder color and higher rehydration time were obtained in this condition. On the other hand, the addition of 20% (w/v) maltodextrin provided better yield. In conclusion, cashew apple juice is a good substrate for the probiotic beverage production, and the condition of drying agents 10% maltodextrin + 10% arabic gum is adequate to maintain satisfactory levels of L. casei NRRL B-442 survival for 35 days, in the powder of probiotic cashew juice stored at 4ÂC. / O objetivo desta pesquisa foi elaborar um produto probiÃtico à base de suco de caju pronto para beber, como tambÃm, na forma desidratada obtida pela secagem por aspersÃo (spray drying). A primeira etapa da pesquisa consistiu em otimizar as condiÃÃes de crescimento do Lactobacillus casei NRRL B-442 em suco de caju, a quantidade adequada de inÃculo e o tempo de fermentaÃÃo. As condiÃÃes Ãtimas para produÃÃo do suco de caju probiÃtico foram: pH inicial de 6,4, temperatura de fermentaÃÃo de 30ÂC, quantidade de inÃculo de 7,48 log UFC/mL (L. casei) e 16 h de fermentaÃÃo. O suco de caju mostrou ser tÃo eficiente quanto os produtos lÃcteos para o crescimento de L. casei. Em uma segunda etapa, foi avaliada a estabilidade da bebida probiÃtica de caju estocada por 42 dias a 4ÂC. Foram realizadas anÃlises no suco de caju nÃo fermentado (controle) e nos sucos fermentados com L. casei NRRL B-442, adicionado ou nÃo de 8% (p/v) de sacarose depois da fermentaÃÃo. Durante a estocagem, foram realizadas as determinaÃÃes de viabilidade de L. casei NRRL B-442, conteÃdo de aÃÃcares e Ãcidos orgÃnicos, cor, atividade antioxidante e enzimÃtica e aceitaÃÃo sensorial. Foi observado que o nÃmero de cÃlulas viÃveis aumentou no suco de caju contendo sacarose ao longo da estocagem. AlÃm disso, a fermentaÃÃo proporcionou um efeito conservante no conteÃdo de Ãcido ascÃrbico que teve uma reduÃÃo menos intensa, com a estocagem, nos sucos fermentados, quando comparados com o controle. A atividade antioxidante e o conteÃdo de polifenÃis apresentaram similar tendÃncia. ReaÃÃes que reduzem o valor nutricional causadas por enzimas foram minimizadas nas amostras fermentadas durante a estocagem. Nessas amostras foi observada maior reduÃÃo da atividade enzimÃtica da polifenoloxidase e peroxidase. Durante a estocagem, o aumento do croma indicou que a cor amarela foi intensificada, sendo bem aceita pelos consumidores. Os atributos sensoriais (aroma, sabor, acidez e cor) do suco de caju probiÃtico foram positivamente influenciados pela estocagem sob refrigeraÃÃo por 42 dias. Na terceira etapa da pesquisa, foi avaliado o efeito da desidrataÃÃo por spray drying no suco de caju contendo L. casei NRRL B-442, alÃm de avaliar a influÃncia da temperatura de estocagem sobre a viabilidade de L. casei e nas propriedades fÃsicas do pÃ, durante 35 dias de estocagem. Os agentes de secagem usados foram: 20% (p/v) de maltodextrina ou 10% (p/v) de maltodextrina + 10% (p/v) de goma arÃbica. O suco de caju probiÃtico desidratado por spray drying apresentou nÃveis satisfatÃrios de sobrevivÃncia de L. casei NRRL B-442, durante a secagem. Durante a estocagem, a adiÃÃo de 10% (p/v) de maltodextrina + 10% (p/v) de goma arÃbica manteve a viabilidade microbiana dentro de nÃveis satisfatÃrios quando o pà foi submetido à refrigeraÃÃo a 4ÂC. Entretanto, maiores diferenÃas na coloraÃÃo do pà reconstituÃdo e maior tempo de reidrataÃÃo foram obtidos nesta condiÃÃo. Jà a adiÃÃo de 20% (p/v) de maltodextrina proporcionou melhor rendimento. Em conclusÃo, o suco de caju pode ser utilizado como substrato para o desenvolvimento de bebida probiÃtica, e a condiÃÃo dos agentes de secagem de 10% de maltodextrina + 10% de goma arÃbica mostra-se adequada para manter os nÃveis satisfatÃrios de L. casei NRRL B-442 por atà 35 dias, no suco de caju probiÃtico desidratado estocado a 4ÂC.

Submitted by Rosangela Silva (rosangela.silva3@unioeste.br) on 2017-08-31T13:37:40Z No. of bitstreams: 2 Andressa Regina Antunes.pdf: 1269867 bytes, checksum: d72378812bab26dd3a8c0c8918a70c93 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-08-31T13:37:40Z (GMT). No. of bitstreams: 2 Andressa Regina Antunes.pdf: 1269867 bytes, checksum: d72378812bab26dd3a8c0c8918a70c93 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-03 / Introduction: The damage caused by the formation of free radicals in the body of animals and humans has presented significative results, once the physiopathology of different types of sickness, such as diabetes, hypertension, atherosclerosis, cancer, among others, follows the binding with the action of these highly reactive components. To reduce the effects caused by them, alternatives in medication are being developed, in order to act as therapeutic adjuvants or prophylactics, like in the case of lactic beverages fermented with probiotic microorganisms. This occurs due to the advantages obtained by both the intrinsic nutrients of these beverages, as well as the microorganisms that forms them, beyond the products of their fermentation. A bacteria species with probiotic properties well-known by their antioxidant potential is the Lactobacillus acidophilus, that is used in fermentation processes for the production of fermented milk, mainly with functional aspects. Objective: Gathering evidences about the antioxidant aspect shown in fermented lactic beverages that contain Lactobacillus acidophilus, by “in vitro” and “in vivo” experimental design tests, besides, evaluating the evidences regarding the other characteristics related to the manufacture of these beverages, like the physical-chemical, microbiologic, antioxidants and methodologic aspects, with the intention of clarifying if these probiotic properties really interfere with the antioxidant aspects of the samples, as well as with the other important aspects for the production of fermented lactic beverages. Material and Methods: A systematic review was done in the data bases: Medline, Cochrane, Scopus, Science Direct, Scifinder, Web of Science, Scielo and Agrícola, considering the following criteria: “antioxidant activity”, “oxidative stress”, “Lactobacillus acidophilus”, “lactic beverage”, “fermented milk”, “yogurt”, “in vitro techniques” and “in vivo”, related with the boolean operators “AND” and “OR”, moreover the manual search for studies on this area that could be of interest. The articles gathered through all the research sources had their titles and abstracts evaluated according to the pre-established inclusion criteria being, basically, the presence of Lactobacillus acidophilus in the constitution of fermented milks, whether isolated or in association with other bacteria, and that had their contents evaluated about their antioxidant aspect. The studies that followed these criteria had their contents fully read. Subsequently, all data of interest were extracted from the included articles, evaluated and compared with literature data for control samples (pure milk) or samples of similar microbiological composition, and also compared with data from brazilian legislation, when available. Results and Discussion: Through the entire data bases searched, 1751 articles were retrieved and read in terms of title and summary. Out of these, 36 articles were selected and fully read, and from them, eight articles followed the inclusion criteria for “in vitro” tests. For “in vivo” tests, six articles were fully read, and considering that only one of them followed the inclusion criteria, it was not possible, therefore, to develop a comparative research for this methodologic design, considering the selected search strategy. Thus, from these eight articles related to “in vitro” experiments, a total of 17 samples of interest were comprehended in the microbiologic constitution requirements, showing at least the presence of Lactobacillus acidophilus, which included three fermented milks (Subgroup A = 17.65%), eight acidophilic milks (Subgroup B = 47.06%) and three yogurts (Subgroup C = 35.29%). From the total declared cellular materials, 66.67% of these were characterized as intracellular content free of cells, being the rest, cellular content without breaking. Regarding the microbiologic constitution, it was possible to verify that, after the intentional presence of Lactobacillus acidophilus, the second most frequent used microorganism was Streptococcus thermophilus, followed by Lactobacillus bulgaricus. Regarding the evaluation of antioxidant activity, the analytic technique used with more frequency was that which evaluates the eliminatory capacity of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, followed by the method of elimination of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical. Based on the profile of antioxidant activity, subgroup B was classified as being the best characterized, due to emcompass, in general, greater number of tests of antioxidant activity in relation to the others. In addition, when constituted by Lactobacillus acidophilus, almost all of the samples evaluated presented consistent results for the antioxidant activity. Conclusion: Based on the obtained results, the importance of association between different analytical methods for evaluation of antioxidant activity is emphasized, due to the greater precision of the recovered results and consequent scientific support given to the study. Besides, it could be verified that, when there is presence of the probiotic of interest in the constitution of the studied food, the antioxidant activity is enhanced, which leads to understand that in fact the microorganism has that characteristic. On the other hand, when in presence of other microorganisms, whether they are probiotic or not, the antioxidant activity is still increased, although not in a proportional way, leading to the understanding that only the presence of Lactobacillus acidophilus is able to provide sufficient antioxidant protection to food in which it is added. Thus, lactic fermented beverages that contain this probiotic could be consumed with the premise of antioxidant protection, mainly by individuals that suffer from sicknesses with physiopathologies related to oxidative stress. / Introdução: O dano causado pela formação de radicais livres no organismo de animais e humanos tem apresentado resultados significativos, uma vez que a fisiopatologia de diversos tipos de doenças, como diabetes, hipertensão, aterosclerose, câncer, entre outras, possui ligação com a ação destes compostos altamente reativos. Para a redução dos malefícios causados por eles, alternativas não medicamentosas estão sendo desenvolvidas, a fim de atuarem como adjuvantes terapêuticos ou profiláticos, como é o caso das bebidas lácteas fermentadas com microrganismos probióticos. Isso ocorre em função das vantagens obtidas tanto pelos nutrientes intrínsecos destas bebidas, bem como dos microrganismos que as constituem, além de seus produtos de fermentação. Uma espécie bacteriana com propriedade probiótica notadamente conhecida por seu potencial antioxidante é o Lactobacillus acidophilus, utilizado em processos fermentativos para a obtenção de leites fermentados, e preferencialmente com aspectos funcionais. Objetivo: Reunir evidências sobre o aspecto antioxidante apresentado por bebidas lácteas fermentadas em presença de Lactobacillus acidophilus, a partir de ensaios com desenhos experimentais “in vitro” e “in vivo”, além de avaliar as evidências a respeito das demais características vinculadas à fabricação destas bebidas, como os aspectos físico-químicos, microbiológicos, antioxidantes e metodológicos, com a intenção de esclarecer se o referido probiótico realmente interfere no aspecto antioxidante das amostras, bem como nos demais aspectos de interesse para a fabricação de leites fermentados. Material e Métodos: Foi realizada uma revisão sistemática nas bases de dados: Medline, Cochrane, Scopus, Science Direct, Scifinder, Web of Science, Scielo e Agrícola, considerando os seguintes termos de busca: “antioxidant activity”, “oxidative stress”, “Lactobacillus acidophilus”, “lactic beverage”, “fermented milk”, “yogurt”, “in vitro techniques” e “in vivo”, associados aos operadores booleanos “AND” e “OR”, além da busca manual por estudos na área que pudessem ser de interesse. Os artigos recuperados por meio de todas as fontes de busca tiveram os seus títulos e resumos avaliados de acordo com os critérios de inclusão preestabelecidos, sendo eles, basicamente, a presença de Lactobacillus acidophilus na constitução de leites fermentados, isolado ou em associação com outras bactérias, e que tiveram os seus conteúdos avaliados quanto ao aspecto antioxidante. Os estudos que atenderam a esses critérios tiveram os seus conteúdos lidos na íntegra. Na sequência, todos os dados de interesse foram extraídos dos artigos incluídos, avaliados e comparados com dados de literatura para amostras controle (leite puro) ou amostras de constituição microbiológica semelhante, e ainda, comparados com dados da legislação brasileira quando disponíveis. Resultados e Discussão: Por meio das bases de dados pesquisadas, 1751 artigos foram recuperados e lidos em termos de título e resumo. Desses, 36 artigos foram selecionados e tiveram seus conteúdos lidos na íntegra, e, a partir deles, oito artigos atenderam aos critérios de inclusão para estudos “in vitro”. Para estudos “in vivo”, seis artigos foram lidos na íntegra, sendo que apenas um deles atendeu aos critérios de inclusão, não sendo possível, portanto, o desenvolvimento de uma pesquisa comparativa para o referido desenho metodológico, considerando a estratégia de busca escolhida. Dessa forma, a partir dos oito artigos envolvendo experimentos “in v vitro”, um total de 17 amostras de interesse se enquadraram no requisito de constituição microbiológica, por apresentarem no mínimo a presença de Lactobacillus acidophilus, as quais incluíram três leites fermentados (Subgrupo A = 17,65%), oito leites acidófilos (Subgrupo B = 47,06%) e seis iogurtes (Subgrupo C = 35,29%). Do total de materiais celulares declarados, 66,67% deles foram caracterizados como conteúdo intracelular livre de células, sendo o restante, o conteúdo celular sem rompimento. Quanto à constituição microbiológica, foi possível verificar que, após a presença intencional de Lactobacillus acidophilus, o segundo microrganismo mais frequentemente utilizado foi Streptococcus thermophilus, seguido por Lactobacillus bulgaricus. No que se refere à avaliação da atividade antioxidante, a técnica analítica empregada com maior frequência foi aquela que avalia a capacidade eliminatória do radical 2,2-diphenyl-1-picrylhydrazyl (DPPH), seguida pelo método de eliminação do radical 2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulphonic acid) (ABTS). Com base no perfil de atividade antioxidante, o subgrupo B foi classificado como sendo o melhor caracterizado, em função de ter englobado, no geral, maior número de testes de atividade antioxidante em relação aos demais. Em adição, quando constituídas por Lactobacillus acidophilus, quase a totalidade das amostras avaliadas apresentaram resultados consistentes para a atividade antioxidante. Conclusão: Partindo dos resultados obtidos, enfatiza-se a importância da associação entre diferentes métodos analíticos para a avaliação da atividade antioxidante, devido à maior precisão dos resultados recuperados e consequente respaldo científico conferido ao estudo. Além disso, verifica-se que, quando a presença do probiótico de interesse se encontra na constituição do alimento estudado, a atividade antioxidante é exaltada, o que leva a entender que de fato o microrganismo apresenta tal característica. Por outro lado, quando em presença de outros microorganismos, probióticos ou não, a atividade antioxidante é ainda melhorada, mas não de forma proporcional, levando à compreensão de que apenas a presença de Lactobacillus acidophilus é capaz de fornecer proteção antioxidante suficiente ao alimento em que for acrescentado. Dessa forma, bebidas lácteas fermentadas em presença desse probiótico poderiam ser consumidas com a premissa de proteção antioxidante, principalmente por indivíduos acometidos pordoenças com fisiopatologias envolvidas com o estresse oxidativo.

La maladie de Crohn (MC) est une maladie inflammatoire chronique du tube digestif caractérisée par un état d’hyperactivation du système immunitaire intestinal. Les données cliniques et expérimentales montrent que l’étiologie de la MC serait une réponse immunitaire aberrante à des facteurs environnementaux et/ou infectieux chez un hôte génétiquement prédisposé. Les patients atteints de MC présentent une perméabilité intestinale anormalement élevée pouvant expliquer la stimulation du système immunitaire intestinal par les bactéries et antigènes du microbiote. La muqueuse iléale des patients atteints de MC est anormalement colonisée par des souches de Escherichia coli ayant la propriété d’adhérer et d’envahir les cellules épithéliales intestinales, de survivre et de se multiplier dans les macrophages en entraînant la sécrétion de TNF-α (Tumor Necrosis Factor-alpha). Un pathovar de E. coli associé à la MC et dénommé AIEC pour « Adherent-Invasive E. coli » a été défini. Les souches AIEC adhérent via l’adhésine FimH des pili de type 1 aux résidus mannose de la glycoprotéine CEACAM6, anormalement exprimée au niveau de l’épithélium iléal des patients atteints de MC. Au cours de la maladie, l’inflammation intestinale peut être contrôlée par les traitements médicamenteux ou la chirurgie sans pour autant obtenir de rémission complète et définitive.Le but du travail était d’analyser les conséquences de l’infection par des bactéries AIEC in vivo dans un modèle murin reproduisant l’interaction AIEC/CEACAM6 et de proposer des stratégies pour éliminer ces bactéries de l’intestin. Nous avons montré que les AIEC avaient la capacité d’altérer la fonction de barrière de l’épithélium intestinal chez des souris transgéniques CEABAC10 exprimant CEACAM6. Cette altération est associée à une forte induction de l’expression de la protéine de jonction Claudine-2, comme observée chez les patients atteints de MC. Nous avons ensuite démontré que la levure S. cerevisiae CNCM I-3856 et des produits de levures étaient capables de maintenir l’intégrité de la barrière intestinale des souris en prévenant la colonisation du tractus digestif par les bactéries AIEC. Dans une deuxième partie, nous nous sommes intéressés au développement de nouvelles molécules antagonistes de l’adhésine FimH. Les thiazolylaminomannosides ont montré un puissant effet inhibiteur de l’adhésion des bactéries AIEC à des cellules épithéliales intestinales. Les heptyl-mannosides sont également de puissants inhibiteurs de l’adhésion des AIEC et leur présentation en multivalence sur des corps polymériques ou des cyclodextrines potentialisent l’effet anti-adhésif. De manière intéressante, certains heptyl-mannosides diminuent fortement la colonisation de l’intestin par les bactéries AIEC et préviennent la colite chez des souris CEABAC10. Toutefois, la multivalence n’apporte pas d’efficacité supplémentaire dans ce contexte. En conclusion, deux stratégies anti-adhésives ont été étudiées : les levures et les mannosides. Elles pourraient être proposées aux patients atteints de MC fortement colonisés par les AIEC afin d’éliminer ces bactéries du tractus digestif pour espérer diminuer l’inflammation. Les perspectives à ce travail seront la mise au point de techniques de détection simples et rapides des personnes colonisées ou susceptibles d’être colonisées par ces souches de E. coli pour cibler la population à traiter par les probiotiques levures et les molécules anti-adhésives. / Crohn’s disease (CD) is an inflammatory bowel disease (IBD) with a multifactorial etiology, resulting from an exacerbated inflammatory response to intestinal microbes and/or microbial components in genetically susceptible hosts. CD patients present an increased intestinal permeability which can favor the overstimulation of the intestinal immune system by bacteria or antigens from microbiota. Ileal mucosa from CD patients is abnormally colonized by Escherichia coli strains sharing the ability to adhere to and to invade intestinal epithelial cells, to survive and to replicate within macrophages, inducing high secretion of TNF-α (Tumor Necrosis Factor-). These strains associated with CD are grouped in a pathovar of E. coli named AIEC for « Adherent-Invasive E. coli ». AIEC bacteria adhere via the adhesin FimH localized at the tip of the type 1 pili, to mannose residues exposed on the glycoprotein CEACAM6 abnormally expressed at the ileal mucosa of CD patients. Currently, intestinal inflammation can be controlled with drugs or intestinal surgery but total remission cannot be yet achieved. The aim of the present work was to investigate consequences of AIEC infection in a murine model mimicking the AIEC/CEACAM6 interaction and to test different strategies to eradicate these bacteria from the gut. We showed that AIEC bacteria altered barrier function of the intestinal epithelium in transgenic CEABAC10 mice expressing human CEACAM6. The overexpression of the pore-forming tight junction protein claudin-2 was correlated with the increase intestinal permeability, as observed in CD patients. We demonstrated that the yeast strain S. cerevisiae CNCM I-3856, as well as some yeast products, were able to prevent increase of intestinal permeability in decreasing AIEC gut colonization. In a second part, we investigated another strategy targeting AIEC bacteria using antagonists to FimH adhesin. Thiazolylaminomannosides molecules exerted a strong inhibitory effect on the ability of AIEC bacteria to adhere to intestinal epithelial cells. Heptyl-mannosides (HM) also shared high inhibitory properties in vitro and their efficacy can be potentiated when HM are harbored in multiple copies on polymeric or cyclodextrin cores. Interestingly, some HM molecules strongly decreased AIEC gut colonization and the signs of colitis in vivo, in AIEC LF82-infected CEABAC10 mice. In that context, multivalency did not improve inhibitors efficacy.To conclude, two different strategies were studied: probiotic yeasts and anti-adhesive mannosides. These treatments should be proposed in CD patients highly colonized by AIEC bacteria in order to eliminate these bacteria from the gut and to decrease intestinal inflammation. Future works will focus on the development of quick and easy detection methods to determine people colonized or susceptible to be colonized by AIEC bacteria to treat this subpopulation of CD patients.

Objetivo: Evaluar el efecto de la incorporación de Camellia sinensis L. (2%, 4% y 6% de extracto de té verde) sobre el contenido de compuesto fenólicos, capacidad antioxidante y aceptación sensorial de un yogurt probiótico. Material y métodos: Estudio experimental de laboratorio dónde se elaboró un yogurt probiótico con té verde añadido en distintas concentraciones de 2%, 4% y 6%. Además, se evaluó la aceptabilidad sensorial y se analizó las características físicoquímicas de cada muestra. Asimismo, para determinar el contenido de polifenoles se utilizó el método de Folin-Ciocalteau y, el método de DPPH (2,2-difenil-1-1-picrilhidrazilo) para determinar la capacidad antioxidante. Resultados: Se encontró que en el yogurt probiótico con 6% de té verde añadido había 1.64 mg/mL de contenido de polifenoles, casi el triple del contenido de polifenoles que en el yogurt probiótico con 2% de té verde añadido. Además, se encontró que el yogurt con mayor porcentaje de té verde añadido (6%) obtuvo 61% de decoloración del reactivo de DPPH, lo que nos indica mayor capacidad antioxidante a comparación de las muestras con menor porcentaje de té verde añadido. Conclusión: A mayor contenido de compuestos fenólicos, mayor es la capacidad antioxidante de un yogurt probiótico con té verde añadido, además de ser el que mayor aceptabilidad sensorial alcanzó, siendo su olor y sabor los atributos más apreciados por los panelistas no entrenados quienes le otorgaron un puntaje de 4 en una escala hedónica de 5 puntos, lo que equivale a un “le gusta moderadamente”. / Objective: To evaluate the effect of Camellia sinensis L. (2%, 4% and 6% of green tea extract) incorporation on the content of phenolic compounds, antioxidant capacity and sensory acceptance of a probiotic yogurt. Material and methods: In this experimental laboratory study, probiotic yogurt with added green tea was elaborated using three concentrations of green tea extract (2%, 4% and 6%), and the sensory acceptability and the physical-chemical characteristics were determined for each concentration. Likewise, to determine the content of polyphenols, the Folin-Ciocalteau method was used, and the DPPH method (2,2-diphenyl-1-1-picrylhydrazyl) was used to determine the antioxidant capacity. Results: It was found that in the probiotic yogurt with 6% green tea extract added there was 1.64 mg / mL of polyphenol content, almost triple the polyphenol content than in the probiotic yogurt with 2% green tea added. In addition, it was found that the yogurt with the highest percentage of green tea added (6%) obtained 61% discoloration of the DPPH reagent, which indicates greater antioxidant capacity compared to samples with a lower percentage of green tea added. Conclusion: The higher the content of phenolic compounds, the greater the antioxidant capacity of a probiotic yogurt with green tea added, besides being the one with the highest sensory acceptability, its smell and taste were the attributes most appreciated by the untrained panelists who gave it a score of 4 on a hedonic scale of 5 points, which is equivalent to a "moderately like". / Tesis

Diarrhoe ist eines der häufigsten Probleme beim equinen Neonaten. Nahezu alle Fohlen entwickeln Durchfall innerhalb der ersten Lebenswochen. Unterschiedliche virale, bakterielle und parasitäre Ursachen werden diskutiert. In diesen Zeitraum fällt ebenfalls die erste Rosse der Stute, sodass der Durchfall um den 5. - 15. Lebenstag (LT) bei den Fohlen als „Fohlenrossedurchfall“ bezeichnet wird. Es wird vermutet, dass die Entwicklung der intestinalen Mikroflora und die Reifung der Darmschleimhaut im Wesentlichen für das Durchfallgeschehen verantwortlich sind. Bisher ist jedoch wenig bekannt über die Entwicklung der intestinalen Mikroflora bei Fohlen. Einige Probiotika sind als Darmflorastabilisatoren bei Tieren zugelassen. Studien belegten positive Effekte von Toyocerin® (B. cereus var. toyoi) auf die Darmgesundheit bei anderen Tierarten wie z.B. Kälbern, Ferkeln, Broilern, Puten und Mastkaninchen. Die vorliegende Arbeit sollte klären, ob die Supplementierung von B. cereus var. toyoi zu einer Stabilisierung der sich entwickelnden intestinalen Mikroflora und damit zu einer Verringerung der Durchfälle bei Fohlen führt. Die Untersuchung wurde an 25 Mutterstuten eines Vollblutgestüts und ihren Fohlen durchgeführt. Alle Fohlen wurden von Februar bis Mai 2011 geboren. Von Geburt an wurden die Fohlen randomisiert in drei Behandlungsgruppen eingeteilt: Placebo-Gruppe (10 ml isotone Kochsalzlösung, n=8), 50 mg Toyocerin-Gruppe (5 x 108 KbE B. cereus var. toyoi gelöst in 10 ml isotoner Kochsalzlösung, n=7) und 200 mg Toyocerin-Gruppe (2 x 109 KbE B. cereus var. toyoi gelöst in 10 ml isotoner Kochsalzlösung, n=10). Die Placebo- und Behandlungsgruppen wurden einmal täglich vom 1. – 58. LT supplementiert. Herz- und Atemfrequenz, Körperinnentemperatur und die Körpermasseentwicklung wurden nach einem standardisierten Protokoll erhoben. Kotproben konnten mit Hilfe von Kotsammelbeuteln oder durch rektale Entnahme innerhalb von 24 Stunden nach der Geburt sowie an LT 9, 16, 23, 30, 44, 58 und am ersten Durchfalltag gewonnen werden. Blutproben wurden aus der Vena jugularis externa am 1., 9., 16., 30., 58. LT sowie am ersten Durchfalltag entnommen. Die bakteriologische Untersuchung erfolgte mit Hilfe des Kulturverfahrens. Die Bestimmung der Gesamt-IgG-Werte wurde mit einem kompetitiven ELISA, die Bestimmung der spezifischen Antikörper IgG-anti-LPS von E. coli J5 und IgG-anti-PLC-von-C. perfringens-1a mit einem indirekten ELISA durchgeführt. 88 % der Fohlen entwickelten Durchfall (Placebo 7/8, 50 mg Toyocerin 5/7, 200 mg Toyocerin 10/10) mit einer hohen Inzidenz zwischen dem 8. und 16. LT. Das Allgemeinbefinden und die Bewegungs- und Sauglust blieben dabei unbeeinflusst. Zum Zeitpunkt des ersten Östrus der Stute zeigten 59 % der Fohlen Diarrhoe. Unter den 41 %, die keinen Durchfall zum Zeitpunkt der Fohlenrosse hatten, waren Fohlen, die nie Durchfall vom 1. – 58. LT zeigten, aber auch welche die Diarrhoe entwickelten, als die Mutterstute sich nicht in Rosse befand. Ein Zusammenhang zwischen der Fohlenrosse der Stute und Durchfall bei ihrem Fohlen konnte nicht hergestellt werden. Es zeigte sich eine Tendenz, dass hohe Spiegel der Gesamt-IgG (>20 mg/ml) und IgG-anti-LPS von E. coli J5 (>120 RE/ml) nach der Kolostrumaufnahme im Zusammenhang mit einer geringeren Anzahl von Durchfalltagen innerhalb der ersten zwei Lebensmonate standen. C. perfringens und Enterobakterien waren gleichermaßen nachweisbar bei Fohlen mit Durchfall als auch bei unauffälligen Fohlen. Aus der Supplementierung von B. cereus var. toyoi ergab sich kein Effekt auf die Kotflora der Fohlen, außer auf die Gesamtkeimzahlen (GKZ) der aeroben Bakterien. Bei den Aerobiern im Fohlenkot konnte ein signifikanter Behandlungseffekt (p=0,012) festgestellt werden. Im ersten Milchkot der Fohlen waren GKZ von 4,5 x 104 KbE/g (200 mg Toyocerin-Gruppe) bis 5,0 x 105 KbE/g (50 mg Toyocerin-Gruppe) bei den aeroben Bakterien und GKZ von 2,4 x 105 KbE/g (200 mg Toyocerin-Gruppe) bis 4,7 x 106 KbE/g (50 mg Toyocerin-Gruppe) median bei den Anaerobiern nachweisbar. Danach stieg der Gehalt der aeroben und anaeroben Bakterien weiter bis zum 3. LT und stagnierte bis zum 16. LT. Während dieser Stagnationsphase trat bei 92 % der Fohlen (23/25) eine Veränderung der Kotkonsistenz bis hin zu Durchfällen auf. Vom 16. bis zum 58. LT sanken die Gehalte moderat bei den Aerobiern median am 58. LT auf 2,7 x 105 KbE/g (Placebo-Gruppe) bis 2,2 x 106 KbE/g (50 mg Toyocerin-Gruppe) und bei den Anaerobiern median am 58. LT auf 3,8 x 105 KbE/g (Placebo-Gruppe) bis 2,9 x 106 KbE/g (200 mg Toyocerin-Gruppe). Bis zum 58. LT näherte sich der Medianwert der aeroben und anaeroben Bakterien im Kot der Placebo-Gruppe dem Wert der Mutterstuten (gemessen am ersten Tag nach der Geburt) an. Innerhalb der ersten Lebenstage war eine hohe aerobe sowie anaerobe Keimzahl im Kot der Fohlen nachzuweisen, die sich oberhalb der Keimzahlen befand, die im Kot der Mutterstuten zum Zeitpunkt der Geburt gemessen wurde. Im Rahmen der Entwicklung und Etablierung der bakteriellen intestinalen Mikroflora wurde das Fohlenrossedurchfallgeschehen bei den Fohlen beobachtet. B. cereus var. toyoi hatte dabei keinen Einfluss auf die Anzahl der Fohlen mit Durchfall und den Gesundheitsstatus der Fohlen. / Diarrhoea is probably one of the most common problems in equine neonates. Almost all foals develop transient diarrhoea within the first weeks of life. Different viral, bacterial and parasitic causes are discussed. Between the 5th and the 15th day of the foal’s life, when their dam’s first post partum (p.p.) oestrus is expected, diarrhoea in foals is observed quite often. That is why it’s called “foal heat diarrhoea”. In literature establishment of intestinal microflora and maturation of the intestinal mucosa is responsible for the occurrence of diarrhoea in this period of life. But little is known about the development of the intestinal microflora in foals. Many probiotics are authorised as gut flora stabilisers in animal nutrition. Some studies proved positive effects of Bacillus (B.) cereus var. toyoi (Toyocerin®) on intestinal health in other species e.g. calves, piglets, broiler chicken, poultry and growing rabbits. The present study deals with the question if a supplementation of B. cereus var. toyoi lead to a stabilisation of the developing intestinal microflora and therefore to a reduction of diarrhoea in foals. A total of 25 mares and foals of a thoroughbred stud were included into the study. Foals were born between February and May 2011. From birth, the foals were randomly assigned to three treatment groups: placebo group (10 ml isotonic saline solution, n=8), 50 mg Toyocerin group (5 x 108 cfu B. cereus var. toyoi solved in 10 ml isotonic saline solution, n=7) and 200 mg Toyocerin group (2 x 109 cfu B. cereus var. toyoi solved in 10 ml isotonic saline solution, n=10). Placebo- and treatment groups were orally supplemented once a day starting on the 1st through to the 58th day of life. Determination of heart and respiratory rate, body temperature, body weight was realised according to a standardised protocol. Within the first day of life, on day 9, 16, 23, 30, 44, 58 and on the first day of diarrhoea faecal samples has been taken from the rectum or by the use of a collection bag. Blood samples were taken via jugular venipuncture on day 1, 9, 16, 30, 58 and on the first day of diarrhoea. Culture-depend methods were used to analyse the bacterial microflora. Serum IgG was analysed by a competitive ELISA, IgG-anti-LPS from E. coli J5 and IgG-anti-PLC-from-C. perfringens-1a by an indirect ELISA. 88 % of the foals developed diarrhoea (placebo 7/8, 50 mg Toyocerin 5/7, 200 mg Toyocerin 10/10) with a high incidence between the 8th and the 16th day of the foal’s life. Meanwhile, foals remained bright and alert and continued to nurse. At the time point of the first p.p. oestrus in the mares, 59 % of their foals showed signs of diarrhoea. Within the remaining 41 % there are foals that had no diarrhoea but there are also foals which had diarrhoea when the mare had not been in heat. Neonatal diarrhoea in foals is not linked to p.p. oestrus in their mares. There was a tendency, that high serum-IgG (> 20 mg/ml) and IgG-anti-LPS from E. coli J5 (> 120 RE/ml) after colostrum uptake were associated with lower diarrhoea severity in the first 58 days of the foal’s life. C. perfringens and enterobacteria can be found equally in foals with diarrhoea and in foals which are not afflicted. B. cereus var. toyoi supplementation had no effect on faecal bacteria in foals, except on aerobic bacteria (p=0,012). In the first milk faeces aerobic bacteria were detected in median from 4,5 x 104 cfu/g (200 mg Toyocerin-group) to 5,0 x 105 cfu/g (50 mg Toyocerin-group) and anaerobic bacteria were detected in median from 2,4 x 105 cfu/g (200 mg Toyocerin-group) to 4,7 x 106 cfu/g (50 mg Toyocerin-group). Afterwards the counts increased towards the 3rd day of life and stayed on a high level till the 16th day of life. During this stagnation in 92 % of the foals a change in faecal consistency and diarrhoea was observed. Afterwards, from the 16th though to the 58th day of life, the bacteria counts in the faeces moderately decreased in median for the aerobic bacteria on the 58th day of life down to 3,8 x 105 cfu/g (placebo-group) till 2,9 x 106 cfu/g (200 mg Toyocerin-group). On the 58th day of life the counts of aerobic and anaerobic bacteria in the faeces of the placebo-group approached the counts in the faeces of the mare (measured at the time point of birth). In the first days of foals’ life detection of aerobic and anaerobic bacteria in the faeces were high, and above the level of the bacteria counts in the faeces of the mare at the time point of birth. Foal heat diarrhoea is observed as a part of the development and establishment of bacterial intestinal microflora. B. cereus var. toyoi had no effect on the percentage of foals with diarrhoea and health status in the foals at that point.

Le microbiote intestinal joue un rôle fondamental chez son hôte. Son étude contribue à la compréhension de différents mécanismes physiologiques chez l’hôte. Suite à l’interdiction d’utiliser des antibiotiques comme facteurs de croissance dans l’alimentation animale, l’utilisation de probiotiques s’est largement répandue pour améliorer les performances zootechniques des animaux. De plus, depuis 1999, des études ont été demandée aux pays producteurs de foie gras de rechercher des méthodes alternatives au gavage. Les travaux de cette thèse se sont donc articulés autour de ces deux objectifs. L’effet de la supplémentation en probiotiques (genre Lactobacillus ou mix de souches) chez des canards mulard durant la phase d’élevage et de gavage a donc été étudié. Si aucun effet des probiotiques sur le métabolisme n’a été démontré, l’expression des marqueurs de l’inflammation a diminué lors du gavage. De plus, nous avons confirmé l’importance du genre Lactobacillus, pendant le gavage, groupe bactérien à fort pouvoir amylolytique. Dans un second temps, la composition du microbiote intestinal de l’oie grise landaise gavée ou suite à une stéatose hépatique spontanée a été étudiée et comparée dans le cadre d’un projet d’alternative au gavage. Pour finir, une étude sur le déclenchement d’une stéatose hépatique chez des oies issues du système de reproduction à l’aide d’un aliment complet de croissance a été réalisée. Ces deux études sur les oies nous ont permis d’observer des similitudes dans les phyla majoritaires du microbiote intestinal qui sont les Firmicutes et les Proteobacteria bien que des différences ont été observées à des niveaux taxonomiques plus bas. L’ensemble de ces travaux ont confirmé la corrélation entre diversité du microbiote intestinal et aptitude à déclencher une stéatose hépatique qu’elle soit issue d’un gavage ou spontanée. / The intestinal microbiota plays a fundamental role in its host. Microbiota studies contribute to understand the different physiological mechanisms in the host. Then, following the ban on antibiotics as growth factors in animal feed, the use of probiotics has increased to improve zootechnical performances. Moreover, since 1999, studies have been requested to the European countries producing “foie gras” to develop alternative methods to overfeeding. The work of this phD was organized according to these two objectives. The effect of probiotic supplementation in ducks during breeding and overfeeding periods was studied. If no impact of probiotics on metabolism has been demonstrated, the expression of markers of inflammation decreased during overfeeding. Moreover, we confirmed the importance of the Lactobacillus genus during overfeeding period, bacterial group with strong amylolytic activities. In a second step, the composition of the intestinal microbiota of the graylag goose after overfeeding period or following a spontaneous hepatic steatosis was studied and compared. Finally, a study on the initiation of hepatic steatosis in graylag geese fed with complete pellets was performed. These two geese studies allowed us to observe the similarities in the major phyla of the intestinal microbiota. Major phylum are still Firmicutes and Proteobacteria although differences were observed at lower taxonomic levels. All of these studies confirmed the correlation between intestinal microbiota diversity and the ability to trigger hepatic steatosis, whether with overfeeding or spontaneous.

Probióticos são suplementos alimentares de microrganismos vivos com efeitos benéficos no hospedeiro animal pela melhora do balanço intestinal. Dentre os microrganismos considerados probióticos, somente aqueles microrganismos classificados como bactérias ácido lácticas (LAB) são considerados importantes em relação à alimentação. Cultivos de altas densidades de células são cada vez mais importantes do ponto de vista industrial para a obtenção de LABs, pois produtos adicionados com esses suplementos apresentam alto valor agregado. Este trabalho tem por objetivo a produção de Lactobacillus plantarum em cultivo submerso em biorreator, avaliação da sua resistência na forma livre e imobilizada frente às condições de armazenamento sob refrigeração e trânsito gastrointestinal e elaboração de produto fermentado com adição de probióticos. Inicialmente foram realizados experimentos para selecionar Lactobacillus que apresentassem alta produtividade de biomassa. Em seguida foram realizados experimentos para avaliar a utilização de soro de queijo como ingrediente base da formulação do meio de cultivo e sua suplementação com diferentes fontes de nitrogênio. Com o microrganismo selecionado, L. plantarum, realizou-se a seleção de variáveis através do delineamento experimental Plackett Burman (P-B). A otimização das condições de cultivo foi realizada utilizando um delineamento composto central rotacional (DCCR). Paralelamente, foram testadas a sobrevivência de L. plantarum em armazenamento sob refrigeração e à exposição a meios que simulem a passagem pelo aparelho digestivo. Estas avaliações foram realizadas comparativamente entre os microrganismos na forma livre e na forma microencapsulada utilizando como polímeros alginato de sódio e pectina e recobrimentos com alginato de sódio e quitosana. Os resultados mostraram que a temperatura, pH, taxa de aeração, concentração de lactose e peptona foram os parâmetros que mais influenciaram a produção de biomassa. O DCCR para temperatura e taxa de aeração mostraram que o máximo de produção de biomassa predita foi de 14,30 g L-1 de L. plantarum, nas condições otimizadas. No ponto central do DCCR, atingiu-se a produção de biomassa de L. plantarum de 10,2 g L-1, como taxa de conversão de 0,10 g de células g-1 de lactose e 1,08 g de ácido láctico g-1 lactose (m/m) com as seguintes condições de cultivos: 140 g L-1 de lactose; 15 g L-1 peptona; 5 g L-1 extrato de levedura; pH 5,2; velocidade de agitação de 200 rpm; 34 ºC e 3,5 vvm. O meio intestinal simulado não interferiu na viabilidade dos microrganismos em relação ao meio controle. Já o meio gástrico simulado diminui drasticamente a viabilidade dos microrganismos nas condições testadas não havendo diferença significativa entre os diferentes materiais imobilizantes utilizados e o controle sem imobilização. No armazenamento sob refrigeração houve aumento da viabilidade em relação às células não imobilizadas, sendo que os tratamentos em que houve menor perda de viabilidade foram imobilização em 4 % de pectina, 3 % de alginato de sódio recoberto com quitosana e mistura de 2 % de alginato de sódio e 2 % de pectina. Quando testada a viabilidade em iogurte de L. plantarum imobilizados em 3 % de alginato recoberto com quitosana houve perda de viabilidade de 0,55 ciclo logarítmico durante 38 dias de armazenamento. A cepa de L. plantarum estudada se mostra como um microrganismos potencial para utilização como probiótico em alimentos, uma vez que demonstrou alta produtividade de células e boa viabilidade frente às condições de estresse utilizadas. / Probiotics are live microorganisms feed supplement, which beneficially affects the host animal by improving its intestinal microbial balance. Among the microorganisms considered probiotics, only those strains classified as latic acid bacteria - LAB are considered of importance regarding to the nutritional effects. High cell density cultivations of LABs are important from the industrial viewpoint, because products added with this supplement are of high value. The aims of this work were to investigate the biomass production of Lactobacillus plantarum in submerged bioreactor cultures, evaluate the resistance of free and immobilized L. plantarum when submitted to refrigerated storage, the viability in simulated gastrointestinal juices and in yoghurt. Initially, experiments were performed to select Lactobacillus that showed high productivity of biomass. Further experiments were performed to evaluate the use of cheese whey as a basic ingredient in the formulation of the medium and its supplementation with different nitrogen sources. The selected microorganism, L. plantarum, was used for the selection of variables of the Plackett Burman (PB) design. The optimization of culture conditions was performed using a central composite rotational (CCD) design. In parallel, it was tested the survival of L. plantarum in refrigerated storage and exposure to media that simulated the passage through the digestive tract. These evaluations were performed comparatively between microorganisms in the free and microencapsulated form using as polymers sodium alginate and pectin, coated with sodium alginate or chitosan. Results have shown that temperature, pH, aeration rate, lactose, and peptone were the most influential over biomass formation. The CCD for temperature and aeration rate showed that the model predicted maximal biomass production of 14.30 g L-1 (dw) of L. plantarum under the optimized conditions. At central point of CCD, it was obtained a biomass production of 10.2 g L-1 (dw), with conversion rates of 0.10 g of cell g-1 lactose and 1.08 g lactic acid g-1 lactose (w/w), with the following conditions: 140 g L-1 of lactose; 15 g L-1 peptone; 5 g L-1 of yeast extract; pH 5.2; stirred agitation of 200 rpm; 34 ºC and 3.5 vvm. The simulated intestinal medium did not affect the viability of microorganisms in relation to the control medium. However, the simulated gastric medium drastically reduces the viability of microorganisms in the conditions tested with no significant difference between the different materials used and the control without immobilization. In refrigerated storage there was an increase in the viability compared to free microorganisms, and the treatments with lower loss of viability were those of 4% pectin, 3% sodium alginate coated with chitosan and a mixture of 2% alginate sodium and 2% pectin. When tested the viability in yogurt of L. plantarum immobilized in 3% alginate coated with chitosan, the viability loss was 0.55 log cycle during 38 days of storage. The strain of L. plantarum studied was shown as a potential organism for use as probiotics in food, since it has shown high yield and good cell viability in the face of stress conditions used.

FONTELES, Thatyane Vidal. Desenvolvimento de uma nova bebida funcional probiótica à base de melão Cantaloupe sonificado. 2011. 122 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Departamento de Tecnologia de Alimentos, Programa de Pós-Graduação em Ciências e Tecnologia de Alimentos, Fortaleza-CE, 2011 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-06-28T14:35:13Z No. of bitstreams: 1 2011_dis_tvfonteles.pdf: 1940091 bytes, checksum: d899f4bcf6d27d83e3c7c5b7f04d65a0 (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-06-28T14:35:32Z (GMT) No. of bitstreams: 1 2011_dis_tvfonteles.pdf: 1940091 bytes, checksum: d899f4bcf6d27d83e3c7c5b7f04d65a0 (MD5) / Made available in DSpace on 2016-06-28T14:35:32Z (GMT). No. of bitstreams: 1 2011_dis_tvfonteles.pdf: 1940091 bytes, checksum: d899f4bcf6d27d83e3c7c5b7f04d65a0 (MD5) Previous issue date: 2011 / The use of fruit juices for the development of functional foods has been an alternative for people who do not want or cannot make use of fermented dairy products. The objective of this research was the development of a drink with probiotic properties trough the fermentation of melon juice by Lactobacillus casei NRRL B-442. The first stage of the work consisted of the optimization of pH and temperature of fermentation for the L. casei cultivation in melon juice. Results showed that this strain has fermentative potential in melon juice, with no need of nutrient supplementation to reach the appropriated values of pH and viable cells. The best operating condition found to guarantee a good growth of the microorganism and ensure the cellular viability necessary for the product to be considered probiotic was the temperature of 31°C with initial pH of 6.1. The technological advantage of using melon juice as a substrate for probiotic fermentation was demonstrated in this work because the natural pH of the juice provided a favorable environment for the probiotic microorganism development. In a second stage, the effect of ultrasound treatment on quality parameters of melon juice was investigated as a function of intensity of ultrasonic energy and treatment time. The ultrasonic processing intensity using 372.93 W/cm2 for 10 minutes improved the quality of the juice reducing the spoilage due to enzyme activity, increasing homogeneity without damaging the product color. In the third phase of the research, the kinetics of probiotic melon juice fermentation and the influence of the product storage under refrigeration on the beverage quality parameters such as survival of L. casei, post-acidification and color were studied. The melon juice was subjected to ultrasonic processing (372.93 W/cm2/10min) and inoculated with 1% (v/v) and incubated at 31°C for 8 hours. The fermented juice was stored at 4°C, and pH, color, cell biomass, and viable cells number were evaluated. Analyses were performed at intervals of 7 days until 42 days of storage. The highest cell count was achieved within 12 hours of study, reaching 9.0 log CFU/mL, but from the 8th of fermentation there was no significant increase in the concentration of viable cells. Thus, 8 hours was elected as the optimal fermentation time to prepare the probiotic melon juice. During refrigerated storage there was acid production and a slight reduction in the number of viable cell. However, the product kept the number of viable cells above the level recommended by the current legislation during 42 days of storage at 4°C, regardless of having been stocked sugar-free or with sugar addition and no damage to the product color was observed. Melon juice was considered a good vehicle for probiotic microorganism as it showed viable cell count higher than 8 log CFU/mL throughout the studied period. / A utilização de sucos de frutas para o desenvolvimento de alimentos funcionais probióticos tem sido uma alternativa de consumo para indivíduos que não querem ou não podem fazer uso de produtos lácteos fermentados. Desse modo, o objetivo deste trabalho foi desenvolver uma bebida com propriedades probióticas à partir da fermentação do suco de melão Cantaloupe por Lactobacillus casei NRRL B-442. A primeira etapa da pesquisa consistiu na otimização de parâmetros de pH e temperatura de fermentação para o desenvolvimento de L. casei no suco de melão. Os resultados mostraram que a linhagem probiótica estudada apresentou potencial fermentativo em suco de melão, não havendo necessidade de suplementação com nutrientes para a obtenção de valores adequados de pH e número de células viáveis. A melhor condição encontrada que garantisse um bom crescimento do micro-organismo além de assegurar a viabilidade celular necessária para que o produto seja considerado probiótico foi a temperatura de 31°C com o pH inicial do suco de 6,1. Neste trabalho foi demonstrada a vantagem tecnológica do uso de suco de melão como substrato para a fermentação probiótica uma vez que o pH natural do suco proporcionou um meio favorável para o desenvolvimento do micro-organismo. Em uma segunda etapa, investigou-se o efeito do tratamento ultra-sônico nos parâmetros de qualidade do suco de melão em função da intensidade de energia ultra-sônica e do tempo de tratamento. O processamento ultra-sônico utilizando intensidade de 372,93 W/cm2 durante 10 minutos melhorou as características de qualidade do suco reduzindo a atividade de enzimas deteriorantes, aumentando a homogeneidade sem causar danos à cor do produto. Considerando estes resultados, na terceira etapa da pesquisa foram estudadas a cinética da produção de suco de melão sonificado probiótico e a influência da estocagem sob refrigeração em parâmetros de qualidade da bebida como: sobrevivência de L. casei, pós-acidificação e cor. O suco de melão foi submetido ao processamento ultra-sônico, inoculado com 1% (v/v) da cultura e incubado a 31ºC por 8 horas. Os sucos foram armazenados sob refrigeração a 4°C. Foram determinados pH e parâmetros de cor da bebida, crescimento celular e o número de células viáveis. As análises foram realizadas em intervalos de 7 dias, até o 42º dia de estocagem. A maior contagem celular foi atingida nas 12 horas do estudo, chegando a 9,0 log UFC/mL, porém a partir de 8ª hora não houve um aumento significativo da concentração de células viáveis, motivo pelo qual elegeu-se 8 horas como o tempo de fermentação ideal para a elaboração do suco de melão probiótico. Durante a estocagem refrigerada, houve produção de ácidos e uma discreta redução do número de células viáveis, porém o produto manteve prevalência de células viáveis (com índices acima do preconizado pela legislação vigente), durante 42 dias de armazenamento a 4°C, independentemente de terem sido estocadas com ou sem açúcar sem prejuízos à coloração do produto. O suco de melão foi considerado um veículo adequado para a ingestão de micro-organismos probióticos, pois apresentou contagem de células viáveis superior a 8 log UFC/mL durante todo o período de estudo.

SANCHO, Soraya de Oliveira. Estudo do potencial de resíduos de frutas tropicais para elaboração de suplemento alimentar probiótico. 2011. 203 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências, Programa de Pós-Graduação em Biotecnologia (Rede nordeste de Biotecnologia), Fortaleza-CE, 2011. / Submitted by demia Maia (demiamlm@gmail.com) on 2016-05-27T16:26:58Z No. of bitstreams: 1 2011_tese_sosancho.pdf: 4193440 bytes, checksum: 7bf111a6bfc02a4e47da8ccf0c7de283 (MD5) / Approved for entry into archive by demia Maia (demiamlm@gmail.com) on 2016-05-27T16:27:24Z (GMT) No. of bitstreams: 1 2011_tese_sosancho.pdf: 4193440 bytes, checksum: 7bf111a6bfc02a4e47da8ccf0c7de283 (MD5) / Made available in DSpace on 2016-05-27T16:27:24Z (GMT). No. of bitstreams: 1 2011_tese_sosancho.pdf: 4193440 bytes, checksum: 7bf111a6bfc02a4e47da8ccf0c7de283 (MD5) Previous issue date: 2011 / The fruit processing industries produce large amounts of residues, which often do not receive adequate attention. This raw material has several constituents beneficial to health, which can be applied in the food industry in the development of new products. This study aimed to determine the feasibility of using agro-industrial residues such food supplements enriched with probiotic. Residues of pineapple (Ananas comosus L. Mer.), acerola (Malpighia glabra L.), cashew apple (Anacardium occidentale L.), guava (Psidium guajava L.), papaya (Carica papaya L.), mango (Mangifera indica L.) and sapodilla (Achras sapota L.) were evaluated. The physicochemical composition of the residues was determined with the analysis of total soluble solids, titratable acidity, pH, ascorbic acid, lipid, moisture, anthocyanins, water activity, protein, ash, total reducing sugars, total phenolics, antioxidant activity and mineral composition (Na, Mn, Ca, Fe, Mg, K, Zn and P) by the microwave digestion, digester block or ash techniques. The samples were analyzed by optical absorption spectrometry with inductively coupled plasma (ICP-OES). Microbial loads of the fruit residues (molds, yeasts, total coliforms and Escherichia coli) were also evaluated. Subchronic toxicity tests were performed with male Swiss mice by administration of extracts from fruit residues (C= 500 mg/ kg orally). The lethality bioassay against Artemia salina L., was conducted at 5,000, 1,000, 500, 100 and 10 mg/ mL concentrations from fruit residue extracts. In addition, tests were performed with the probiotic Lactobacillus casei NRRL B-442 grown in skim milk 10%, submitted to the lyophilization or atomization processes. The analytical results showed rich nutritional properties of the fruit residues and the acerola residue showed the highest values of total anthocyanins (60.83 mg/ 100 g), total phenolic compounds (173.30 mg EAG/ 100 g), ascorbic acid (170.73 mg/ 100 g) and antioxidant activity (48.21%). The microwave digestion procedure is the most suitable for minerals determination of fruit residues given the easiest and simplest analysis. The subchronic toxicity study, in the experimental conditions evaluated did not cause mortality or toxicity in male mice; however, additional studies are recommended. The lethality bioassay with Artemia salina L. indicated that acerola and cashew apple extracts showed biological activity, with LC50 values of 209.90 and 148.10 mg/ mL, respectively. In this study, no fungal contamination was detected, only the guava residue showed total coliforms and there was no evidence of E. coli in all samples, indicating no contamination of fecal origin. In tests with Lactobacillus casei, all counts obtained in fermented milk samples, submitted to drying processes, were within the limits required by legislation to be considered probiotic. Due to the nutritional properties, coupled with functional compounds and higher antioxidant activity, the acerola residue is proposed for the preparation of food supplement enriched with probiotic / As indústrias de processamento de frutas produzem uma grande quantidade de resíduos, que muitas vezes não recebem atenção adequada. Esta matéria-prima apresenta vários constituintes benéficos à saúde, que podem ser aplicados na indústria alimentícia no desenvolvimento de novos produtos. O presente trabalho teve como objetivo determinar a viabilidade de utilização de resíduos agroindustriais para a elaboração de suplemento alimentar enriquecido com probiótico. Foram analisados os resíduos de abacaxi (Ananas comosus L. Mer.), acerola (Malpighia glabra L.), caju (Anacardium occidentale L.), goiaba (Psidium guajava L.), mamão (Carica papaya L.), manga (Mangifera indica L.) e sapoti (Achras sapota L.). A composição físico-química dos resíduos foi determinada com as análises de sólidos solúveis totais, acidez total titulável, pH, ácido ascórbico, lipídios, umidade, antocianinas, atividade de água, proteínas, cinzas, açúcares redutores totais, compostos fenólicos totais, atividade antioxidante, além da composição de minerais (Na, Mn, Ca, Fe, Mg, K, Zn e P), a partir das técnicas de digestão por microondas, bloco digestor e cinzas. As amostras foram analisadas através de espectrometria de absorção óptica com plasma indutivamente acoplado (ICP-OES). Além disto, foram realizados os testes microbiológicos de bolores, leveduras, coliformes totais e Escherichia coli. Ensaios de toxicidade subcrônica foram realizados em camundongos Swiss machos, pela administração de soluções de extratos de resíduos de frutas (C= 500 mg/ kg, via oral). O bioensaio de letalidade frente à Artemia salina L., foi realizado em concentrações de 5.000, 1.000, 500, 100 e 10 µg/ mL de extratos de resíduos de frutas. Além disto, foram realizados testes com a bactéria Lactobacillus casei cultivada em leite desnatado 10%, submetido aos processos de secagem por liofilização e atomização. Os resultados analíticos indicaram ricas propriedades nutricionais em resíduos de frutas e o resíduo de acerola apresentou os maiores valores de antocianinas totais (60,83 mg/ 100 g), compostos fenólicos totais (173,30 mg EAG/ 100 g), ácido ascórbico (170,73 mg/ 100 g) e atividade antioxidante (48,21%). O processo de digestão por microondas foi o mais indicado para a determinação de minerais, devido à sensibilidade e facilidade de análise. O estudo de toxicidade subcrônica, nas condições experimentais utilizadas, não resultou em mortalidade ou toxicidade em camundongos machos, entretanto, estudos adicionais são recomendados. Os resultados para o bioensaio com Artemia salina L. indicaram que os extratos de acerola e caju apresentaram atividade biológica, com valores de LC50 iguais a 209,90 e 148,10 µg/ mL, respectivamente. Neste estudo, não foi detectada contaminação por bolores e leveduras; apenas a amostra de goiaba apresentou contaminação por coliformes totais e não houve evidência de E. coli em nenhuma amostra, indicando ausência de contaminação de origem fecal. Nos ensaios com Lactobacillus casei, todas as contagens obtidas nas amostras de leite fermentado, submetidos aos processos de secagem, se encontravam dentro do limite exigido pela legislação para ser considerado probiótico, podendo ser utilizado para a elaboração de suplementos alimentares. Devido às propriedades nutricionais, aliado aos compostos funcionais e atividade antioxidante mais elevada, o resíduo de acerola é proposto para elaboração de suplemento alimentar enriquecido com probiótico.