O objetivo deste estudo foi avaliar o efeito da suplementação com óleos essenciais e levedura viva Saccharomyces cerevisiae na dieta de equinos. Foi avaliada a digestibilidade aparente dos nutrientes da dieta, resposta glicêmica e insulinêmica pós-prandial, perfil plasmático de triglicerídeos, colesterol total, frações de colesterol ligado à lipoproteína de alta densidade (HDL-C), colesterol ligado à lipoproteína de baixa densidade (LDL-C), colesterol ligado à lipoproteína de muito baixa densidade (VLDL-C), pH fecal e população microbiana das fezes. Foram utilizados oito equinos da raça Mini-horse, machos, castrados, com idade de 48±6 meses e peso inicial de 147±15 kg, alimentados com dieta contendo 60% de concentrado comercial multiparticulado e 40% de feno de gramínea. Os tratamentos foram divididos em controle (sem adição de óleo essencial e levedura), levedura viva Saccharomyces cerevisiae (2 g/dia de produto comercial com concentração de 20x109 UFC/g de levedura), óleos essenciais (150 mg/dia de composto comercial de óleos essencias) e associação de Saccharomyces cerevisiae com óleos essenciais nas mesmas quantidades já mencionadas. O delineamento experimental utilizado foi em quadrado latino duplo 4x4 com medidas repetidas no tempo e os dados obtidos foram submetidos à análise de variância com nível de significância de 5%. Não foi observado efeito de tratamento (P>0,05) para os diferentes coeficientes de digestibilidade de matéria seca (63,92%), matéria orgânica (66,70%), proteína bruta (82,78%), extrato etéreo (71,38%), amido (95,24%), fibra em detergente neutro (47,50%) e fibra em detergente ácido (36,40%). Na avaliação da Área abaixo da curva (AAC), não foi observado efeito de tratamento (P>0,05) para glicose (579,75 mg/dL) e insulina (858,38 µU/dL). Na avaliação de gordura plasmática não foi encontrado efeito de tratamento (P>0,05) para triglicérides (43,79 mg/dL), colesterol total (111,21 mg/dL) e frações HDL (66,37 mg/dL), LDL (36,1 mg/dL) e VLDL (8,78 mg/dL). O valor de pH fecal observado (6,42) foi adequado para o equilíbrio da microbiota e não houve diferença entre os tratamentos (P=0,56). Avaliando-se a população microbiana das fezes não foi possível observar efeito para Fibrobacter succinogenes (P=0,08). Houve interação entre óleo e levedura para Ruminococcus flavefaciens (P=0,03). Observou-se também efeito de levedura para Lactobacillus genus (P=0,04). A inclusão de óleos essenciais e levedura Saccharomyces cerevisiae na dieta de equinos não tem efeito sobre a digestibilidade aparente dos nutrientes, resposta glicêmica e insulinêmica, perfil plasmático de triglicerídeos, colesterol e suas frações e pH fecal, porém a inclusão de levedura viva provoca alteração na população microbiana / The aim of this study was to evaluate the effect of supplementation of essential oils and live yeast Saccharomyces cerevisiae on the diet for horses. It was assessed apparent digestibility of nutrients, glucose and insulin postprandial responses, plasma levels of triglycerides, total cholesterol and its fractions high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), very low density lipoprotein (VLDL-C), fecal pH and microbial population in faeces. Eight horses were used, Mini-horse breed, geldings, aged 48±6 months, initial weight 147±15 kg, fed a diet containing high concentrate, consisting of 60% commercial concentrate and 40 % of grass hay. Treatments were divided in control (without essential oils and live yeast), live yeast Saccharomyces cerevisiae (2g/day of a commercial product with a concentration of 20x109 CFU/g of yeast), essential oils (150mg/day of a commercial product consisting of essential oils) and association of Saccharomyces cerevisiae with essential oils in the same quantity mentioned. The experimental design was 4x4 double Latin square with repeated measures over time and the data were subjected to analysis of variance with a significance level of 5%. It was not observed treatment effect (P>0.05) for the different digestibility coefficients of dry matter (63.92%), organic matter (66.70%), crude protein (82.78%), ether extract (71.38%), starch (95.24%), neutral detergent fiber (47.50%) and acid detergent fiber (36.40%). In evaluation of the area under the curve (AUC), wasn't observed effects of treatment (P>0.05) for values of glucose (579.75 mg/dL) and insulin (858.38 µU/dL). In the evaluation of plasma lipids weren't found effect of treatment (P>0.05) for values of triglyceride (43.79 mg/dL), total cholesterol (111.21 mg/dL) and fractions HDL (66.37 mg/dL), LDL (36.1 mg/dL) e VLDL (8.78 mg/dL). Fecal pH value (6.42) was found suitable to the balance of microorganisms, but there weren’t effect of treatment (P=0.56). In quantification of microbial population weren’t observed effect for Fibrobacter succinogenes (P=0.08), but were found effect for interaction of essential oils and live yeast for Ruminococcus flavefaciens (P=0.03) and effect for Lactobacillus genus with the inclusion of yeast (P=0.04). The inclusion of essential oil and live yeast has no effect on digestibility of nutrients, glycemic and insulinemic response, plasma levels lipids and pH of faeces, however the inclusion of live yeast causes alteration in the microbial population

O presente trabalho teve como objetivo verificar a viabilidade da cepa probiótica Bifidobacterium animalis subsp. lactis Bb-12 incorporado em margarina, suplementada com inulina, concentrado protéico de soro (WPC) e concentrado de caseína (CMP), bem como avaliar as características do produto e a resistência do probiótico às condições simuladas do trato gastrintestinal humano. Foram produzidos 7 diferentes tipos de margarinas de mesa (60% de lipídios: 60 % de óleo de palma + 40% de óleo de canola), empregando-se um modelo de mistura, onde inulina, WPC e CMP foram as variáveis estudadas. Uma formulação controle foi produzida (M8), sem adição desses ingredientes. A utilização da mistura do óleo de palma com óleo de canola favoreceu nutricionalmente as formulações, fornecendo produtos contendo ácidos graxos essenciais em sua composição e ausência de ácidos graxos trans. As formulações M1 a M7, exceto a formulação M2 após o 21º dia de armazenamento, apresentaram populações satisfatórias de Bb-12 para um alimento probiótico, com populações acima de 6 log UFC/g durante 35 dias de armazenamento. Margarinas suplementadas com inulina apresentaram populações satisfatórias durante todo o armazenamento, atingindo populações de 8,01 log UFC/g ao 35º dia (M1). Além disso, M3 e M6, revelaram populações de Bb-12 de 6,87 log UFC/g e 7,27 log UFC/g (dia 35), respectivamente. Por outro lado, M8 não foi caracterizada como margarina probiótica, uma vez que apresentou populações abaixo de 6 log UFC/g, já ao 1º dia de armazenamento. Embora WPC seja utilizado em pesquisas para aumentar a viabilidade de probióticos em alimentos, a suplementação de margarina com WPC sem inulina ou CMP não resultou em populações satisfatórias de Bb-12, apresentando decréscimo de 7,82 (dia 1) para 4,64 log UFC/g (M2, dia 35) (p<0,05). Durante todo o ensaio de resistência in vitro, Bb-12 apresentou sobrevivência significativamente superior (p<0,05) em M1 e revelou populações acima de 6 log UFC/g após 6h de ensaio mesmo ao 28º dia. As populações observadas para M2 diminuíram drasticamente durante o ensaio in vitro (5 log UFC/g após 2h no dia 7). Para as outras formulações, as populações de Bb-12 diminuíram 2 log UFC/g após 2h de ensaio in vitro. Entretanto, M1, M2 e M5 (dias 14 e 28) revelaram aumento significativo nas populações de Bb-12 (p<0,05) entre a fase gástrica (2h) e a segunda fase entérica (6h). As margarinas suplementadas com inulina, principalmente M1, revelaram decréscimo significativo no pH durante todo o armazenamento (p<0,05). Entretanto, isto não afetou a qualidade sensorial dos produtos, uma vez que não foram detectadas diferenças significativas entre as formulações após 7 e 14 dias de armazenamento (p>0,05). A suplementação de margarina com inulina e CMP garantiu populações apropriadas de Bb-12 durante o armazenamento estudado pelo menos até o 28º dia. Além disso, contribuiu para sua sobrevivência durante o ensaio de resistência in vitro. Os resultados revelaram que a margarina apresenta-se como uma matriz alimentar adequada para administração de Bb-12, principalmente quando a inulina foi adicionada. / This study aimed to determine the viability of probiotic Bifidobacterium animalis subsp. lactis Bb-12 incorporated in margarine, with inulin, whey protein concentrate (WPC) and caseinomacropeptide (CMP) supplementation. In addition, the in vitro resistance of Bb-12 incorporated in margarine and related properties were evaluated. Seven margarine-making trials (60% of fat: 60% of palm oil +40% canola oil) were produced, using a mixture model, where inulin, WPC and CMP were the variables studied. Also, a control formulation without these ingredients was manufactured. The use of blending palm oil with canola oil improved the margarine formulations nutritionally, providing products containing essential fatty acids in its composition and absence of trans fatty acids. The formulations M1 to M7, except M2 after 21 days of storage, revealed satisfactory Bb-12 populations for a probiotic food, with counts above 6 log CFU/g during 35 days of storage at 5±1ºC. Margarines supplemented with inulin presented suitable Bb-12 populations throughout the whole storage period, reaching up to 8 log CFU/g by the end of storage (M1). Also, M3 and M6, revealed Bb-12 populations of 6.87 log CFU/g and of 7.27 log CFU/g (day 35), respectively. In contrast, M8 was not characterized as probiotic margarine, since it showed Bb-12 populations below 6 log CFU/g on day 1. Even though whey protein is largely employed in probiotic foods, margarine supplementation with WPC without inulin or CMP did not lead to Bb-12 satisfactory populations, decreasing from 7.82 (day 1) to 4.64 log CFU/g (M2, day 35) (p<0.05). During the whole in vitro assays, Bb-12 survived significantly better (p<0.05) in M1 and revealed populations above 6 log CFU/g after 6h even after 28 days. M2 populations decreased drastically during the in vitro assays for all storage period tested (reduction of 5 log CFU/g after 2h of in vitro assays on day 7 and populations of 2.8 log CFU/g after 6h). For the other formulations, Bb-12 populations decreased 2 log CFU/g after 2h of the in vitro assays. However, for M1, M2 and M5 (on day 14 and 28) the populations of Bb-12 increased significantly (p<0.05) between the gastric phase (2h) and the enteric phase (6h). Formulations containing inulin, mainly M1, showed a significant decrease in pH values during the whole storage period (p<0.05). However, this ingredient did not affect the sensory quality of products, since no significant differences between formulations after 7 and 14 days of storage were observed (p>0.05). The supplementation of margarine with inulin and CMP guaranteed appropriate Bb-12 populations during storage for at least 28 days, and also contributed for its survival throughout the in vitro assays. Therefore, margarine might be considered an appropriate food matrix for Bb-12 survival, mainly when inulin is also added.

Além do aspecto nutricional de suma importância, é notória a contribuição do leite humano para o processo de desenvolvimento da microbiota intestinal do recémnascido, um importante mecanismo de defesa do organismo contra doenças infecciosas. O papel do leite humano como fonte de bactérias probióticas, principais constituintes da microbiota intestinal, tem sido tópico de pesquisas recentes. Este trabalho foi desenvolvido com o objetivo de determinar e comparar a composição da microbiota de oito amostras de leite humano e verificar o potencial de utilização desse produto como fonte de bactérias probióticas. Para tanto, utilizaram-se cinco meios de cultivos seletivos para contagem presuntiva de gêneros normalmente encontrados em leite humano: lactococos, enterococos, bifidobactérias e propionibactérias. A análise quantitativa da microbiota demonstrou tendência de diminuição da contagem em função do aumento do tempo de lactação. A análise qualitativa confirmou a presença de distintos gêneros de bactérias lácticas potencialmente probióticas com algumas variações entre as amostras de leite humano. Na segunda etapa 800 colônias isoladas a partir dos cinco meios de cultivos e caracterizadas como bactérias lácticas foram selecionadas quanto às suas propriedades probióticas (produção de bacteriocina, tolerância à acidez e a sais biliares, resistência à antibióticos, capacidade de adesão a chapas de aço inoxidável) e tecnológicas (capacidade de crescimento e sobrevivência em leite). Verificou-se que apenas 15 (1,9%) linhagens produziram bacteriocinas com atividade contra Listeria innocua L11 e Micrococcus luteus ATCC®4698, linhagens utilizadas como indicadoras, por meio do método de antagonismo simultâneo em poços, usando ágar MRS. Treze dessas linhagens também apresentaram atividade contra Bacillus cereus CTC 011, Listeria monocytogenes ATCC®7644, Lactococcus lactis subsp. lactis CTC 204 e Lactobacillus helveticus ATCC®15009. As duas linhagens remanescentes demonstraram atividade principalmente contra Listeria monocytogenes ATCC®7644. Nenhuma das quinze culturas produtoras de bacteriocinas apresentou atividade contra as bactérias Gram-negativas Escherichia coli ATCC® 2074 e Salmonella thyphimuirim ATCC® 2364. Por outro lado, Staphylococcus aureus ATCC® 1602 foi resistente as quinze bacteriocinas selecionadas neste trabalho. Seis linhagens de bactérias lácticas (BALs) foram selecionadas para avaliação das demais propriedades probióticas. Observou-se que uma dessas linhagens diferenciou-se por apresentar sobrevivência a pH 2,0 e a pH 3,0, enquanto as demais mostraram tolerância apenas a pH 3,0. Todas as linhagens selecionadas apresentaram a capacidade de tolerância a 0,3% de sais biliares, de se aderir à superfície de aço inoxidável e de resistência à clindamicina, eritromicina e gentamicina. Quanto às propriedades tecnológicas, todas as seis linhagens apresentaram capacidade de crescimento em leite e não produziram odor desagradável ou pós-acidificação do leite fermentado durante a estocagem a 4ºC por 28 dias. Notou-se, entretanto, diminuição, de, aproximadamente, 2,0 Log UFC.mL-1, na contagem de células viáveis ao final do período de estocagem. Finalmente, por meio da avaliação dos perfis de fermentação de carboidratos e de outras reações bioquímicas, duas das linhagens isoladas de leite humano foram identificadas como Enterococcus durans e quatro como Enterococcus avium. Os 12 resultados permitem concluir que o leite humano é fonte potencial de bactérias com potencial probiótico para aplicação industrial. / In addition to the nutritional aspect of paramount importance, it is clear the contribution of human milk for the development process of the intestinal tract of the newborn, an important mechanism of defense against infectious diseases. The role of human milk as a source of probiotic bacteria, major constituents of the intestinal microbiota, has been the topic of recent research. This work was carried out to determine and compare the composition of the microbiota of eight human milk samples and verify the potential use of this product as a source of probiotic bacteria. For this purpose, we used five selective culture media for counts of presumptive genera commonly found in human milk: lactococcal, enterococci, bifidobacteria and propionibactérias. The quantitative analysis of microbes showed a trend of decreasing counts as a function of time increased lactation. The qualitative analysis confirmed the presence of different kinds of potentially probiotic lactic acid bacteria with some variations between samples of human milk. In the second stage 800 colonies isolated from the five culture media and characterized as lactic acid bacteria were selected for their probiotic properties (bacteriocin production, tolerance to acid and bile salts, antibiotic resistance, adhesion to stainless steel plates) and technology (ability to grow and survive in milk). It was found that only 15 (1.9%) produced bacteriocin strains with activity against Listeria innocua L11 and Micrococcus luteus ATCC 4698®, strains used as an indicator, by the method of antagonism wells simultaneously, using MRS agar. Thirteen of these strains also showed activity against Bacillus cereus CTC 011, Listeria monocytogenes ATCC® 7644, Lactococcus lactis subsp. CTC 204 lactis and Lactobacillus helveticus ATCC 15009®. The two remaining strains showed activity mainly against Listeria monocytogenes ATCC 7644®. None of the fifteen cultures producing bacteriocins active against Gram-negative Escherichia coli ATCC 2074® and Salmonella thyphimuirim ATCC 2364®. Moreover, Staphylococcus aureus ATCC 1602® was resistant the fifteen bacteriocins selected in this work. Six strains of lactic acid bacteria (BALs) were selected for analysis of other probiotic properties. It was observed that one of these strains distinguished by presenting survival at pH 2.0 and pH 3.0 while the other showed only tolerance to pH 3.0. All strains were selected for the ability tolerance of 0.3% bile salts, of adhering to stainless steel surface and resistance to clindamycin, erythromycin and gentamycin. The technological properties, all six strains were capable of growth in milk and produced no unpleasant odor or post-acidification of the fermented milk during storage at 4°C for 28 days. It was noted, however, decreased from approximately 2,0 Log UFC.mL-1 in the viable cell count at the end of storage period. Finally, by evaluating the profiles of fermentation of carbohydrates and other biochemical reactions, two of the strains isolated from human milk have been identified as Enterococcus durans and Enterococcus avium as four. The results indicate that human milk is a potential source of probiotic bacteria with potential for industrial application.

Orientador: José de Assis Fonseca Faria / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-24T19:36:36Z (GMT). No. of bitstreams: 1 Jesus_AnaLauraTiberiode_M.pdf: 1612991 bytes, checksum: 3935cc6ba87a724d5aaf02c88338eea5 (MD5) Previous issue date: 2014 / Resumo: O sódio é um elemento essencial na alimentação, contudo, o seu consumo em excesso poderá ter efeitos nocivos para a saúde, tais como o aumento da pressão sanguínea e a redução da absorção de cálcio. Devido a estes efeitos, a redução do teor de sódio nos alimentos tem sido uma das prioridades nas campanhas realizadas por órgãos governamentais, nomeadamente no setor de laticínios onde o queijo é considerado um gênero alimentício com elevados níveis deste elemento. Dessa forma, o presente trabalho objetivou avaliar o efeito da substituição parcial de NaCl por KCl e MgCl2 nos parâmetros de qualidade do queijo cottage probiótico durante 28 dias de estocagem refrigerada e verificar a viabilidade da cultura starter e das culturas probióticas utilizadas para o processamento do queijo. Além disso, objetivou-se avaliar a aceitação sensorial para identificar direções para a melhoria de atributos sensoriais do produto perante consumidores. Com base nos resultados obtidos observou-se que o ideal de gosto salgado para o queijo cottage probiótico com NaCl foi de 1,12% (p/p), e as concentrações equivalentes em salinidade para todos os sais hipossódicos avaliados mantiveram-se as mesmas do NaCl (1,12% p/p). As contagens de Lactobacillus acidophilus e Bifidobacterium animalis subsp. lactis variaram de 7,74 a 6,20 e 8,58 a 7,93 log (UFC. g-1), respectivamente no primeiro e no último dia de estocagem, valores esses suficientes para benefícios à saúde humana. Porém, as contagens da cultura starter permaneceram baixas com valores inferiores a 3,0 log UFC. g-1 para todas as formulações. Com relação aos ácidos orgânicos, o ácido lático foi o mais encontrado em todas as formulações, seguido pelo ácido cítrico e ácido acético. Quanto a atividade proteolítica, os resultados demonstraram que a formulação controle (100% p/p NaCl sem probióticos) foi a que apresentou menor atividade durante 28 dias de estocagem refrigerada, diferindo estatisticamente das demais formulações. Todos os queijos desse estudo podem ser classificados como de baixo teor de gordura, porque o teor de gordura variou de 4.33 a 4.69 %, e muito alta umidade (74.60 a 75.59 %), segundo a legislação brasileira. A adição de culturas probióticas e a redução de sódio não provocaram mudanças significativas no perfil de ácidos graxos em todos os queijos formulados. Os resultados demonstraram que todas as formulações de queijo cottage apresentaram potencial probiótico, desde que consumidas numa quantidade adequada (2 porções diárias de 50 g), uma vez que as concentrações dos probióticos permaneceram dentro dos padrões estipulados pela legislação brasileira. Entre as formulações elaboradas com sais hipossódicos, a formulação mais aceita foi aquela com redução de sódio de 25% (p/p) de NaCl e substituição por KCl (12,5% p/p) e MgCl2 (12,5% p/p), e o atributo que mais afetou a aceitação dessas formulações foi a textura das amostras. Dessa forma, o queijo cottage probiótico demonstrou ser uma excelente matriz para inserção de sais-hipossódicos, pois não modifica o poder de salga em relação ao NaCl / Abstract: Sodium intake is essential for the welfare of human being, however, its overconsumption may lead to unhealthy effects, namely increased blood pressure and lower calcium absorption. Due to the aforementioned effects, sodium content in food is one of the main focuses of attention in campaigns by government agencies, especially in the dairy sector where cheese is perceived as being a food with high sodium content. Thus, this study aimed to evaluate the effect of partial replacement of NaCl by KCl and MgCl2 on the parameters of quality probiotic cottage cheese during 28 days of refrigerated storage and to determine the viability of starter culture and probiotic cultures used in cheese processing. Furthermore, the objective was to evaluate the sensory acceptance to identify directions for improving the sensory attributes of the product before consumers.Using magnitude estimation method, the salty flavor in probiotic cottage was generated using 1.12% (w/w) NaCl, and equivalent concentrations in salinity for all low-sodium salts, was the same as for NaCl (1.12% w/w). The counts of Lactobacillus acidophilus and Bifidobacterium animalis subsp. lactis ranged from 7.74 to 6.20 and from 8.58 to 7.93 log (CFU. g-1), respectively in the first and last day of storage, enough for benefits to consumers. However, the starter culture counts remained at values below 3.0 log CFU. g-1 for all formulations. Lactic acid was the organic acid found in greater quantities in all formulations, followed by citric acid and acetic acid. The proteolytic activity results showed that the control formulation (100% w/w NaCl without probiotics) showed the lowest activity during 28 days of refrigerated storage, differing from the other formulations. All cheeses of this study can be classified as low-fat, because the fat content ranged 4.33 to 4.69% and very high humidity (74.6 to 75.59%), according to Brazilian regulation. The addition of probiotic cultures and reducing sodium did not cause significant changes in the fatty acid profile of all tested cheeses.The results showed that all the formulations showed probiotic potential if consumed in appropriate portion (2 portions of 50 g daily), since the concentration of probiotics remained within the Brazilian standards. Among formulations prepared with low-sodium salt the best one was that with a reduction of 25%(w/w) sodium substitution of KCl and NaCl (12.5% w/w) and MgCl2 (12.5% w/w) and the attribute that most affected the acceptance was the texture of the samples. Thus, the probiotic cottage cheese proved to be an excellent matrix for the insertion of low-sodium salts, because it does not change the power of brining in relation to NaCl / Mestrado / Tecnologia de Alimentos / Mestra em Tecnologia de Alimentos

Orientador: Mirna Lúcia Gigante / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-24T21:10:27Z (GMT). No. of bitstreams: 1 Chaves_KarinadaSilva_D.pdf: 1644901 bytes, checksum: fb62d45480675d864d6730df243f1fb3 (MD5) Previous issue date: 2014 / Resumo: A adição do probiótico na fabricação de queijos altera a sua microbiota e, consequentemente, pode afetar o desenvolvimento da maturação, o perfil sensorial e funcional do produto. O objetivo desse trabalho foi avaliar o desenvolvimento da maturação, a viabilidade dos micro-organismos e as características físico-químicas e sensoriais de queijo Prato probiótico tipo lanche adicionado de Lactobacillus acidophilus La-5 e Bifidobacterium Bb-12, separadamente ou juntos. Avaliou-se também, o efeito da matriz queijo sobre a viabilidade dos micro-organismos probióticos durante a simulação da passagem pelo trato gastrointestinal. Para a fabricação dos queijos foram realizados os seguintes tratamentos: 1) adicionado de cultura láctica tipo O (queijo controle); 2) adicionado de cultura láctica tipo O e L. acidophilus; 3) adicionado de cultura láctica tipo O e Bifidobacterium; 4) adicionado de cultura láctica tipo O, L. acidophilus e Bifidobacterium. O experimento foi realizado em esquema fatorial 4 x 6, em blocos inteiramente casualizados com três repetições. Os queijos foram avaliados quanto à composição físico-química, proteólise, textura e viabilidade dos micro-organismos após 1, 7, 14, 28, 40 e 60 dias de armazenamento refrigerado. A viabilidade dos micro-organismos probióticos durante simulação das condições gastrointestinais foi avaliada após 28 e 60 dias de armazenamento. Os resultados foram avaliados por Análise de Variância (ANOVA) e teste de Tukey para comparação de médias a um nível de significância de 5%. A análise sensorial descritiva através do método de Perfil Livre dos queijos foi realizada após 28 e 60 dias de armazenamento e os resultados foram avaliados por Análise de Procrustes Generalizada (APG). Os resultados indicaram que os queijos controle e probióticos apresentaram composição típica de queijo Prato e que os tratamentos não afetaram sua composição. Durante 60 dias de armazenamento, os queijos apresentaram aumento da proteólise, redução da firmeza e os queijos probióticos mantiveram altas contagens de L. acidophilus e Bifidobacterium. Os queijos probióticos apresentaram perfis sensoriais similares, com características de aroma e sabor de queijo Prato mais pronunciados que o queijo controle. Os micro-organismos probióticos adicionados nos queijos foram resistentes à simulação do trato gastrointestinal e à bile. O conjunto dos resultados indicou que a adição de probióticos em queijo Prato mostrou-se uma alternativa viável, uma vez que promoveu a manutenção da viabilidade destes micro-organismos durante o processamento e armazenamento refrigerado sem ocasionar alterações indesejáveis nas características sensoriais do produto e conferiu proteção aos probióticos durante a simulação gastrointestinal / Abstract: The addition of probiotics during cheese manufacture changes its microbiota and, consequently, may affect ripening, sensory and functional profile of the product. The aim of this study was to evaluate ripening, viability of microorganism, physicochemical and sensory characteristics of probiotic Prato cheese with L. acidophilus La-5 and Bifidobacterium Bb-12 added separately or together. The effect of the cheese matrix on the viability of the probiotics during the simulation of the passage through the gastrointestinal tract was also evaluated. For the manufacture of the Prato cheeses, the following treatments were studied: 1) addition of type O lactic culture (control cheese); 2) addition of type O lactic culture and L. acidophilus; 3) addition of type O lactic culture and Bifidobacterium; 4) addition of type O lactic culture, L. acidophilus, and Bifidobacterium. The experiment was repeated three times, and assembled in a 4 x 6 factorial design, using a completely randomized block design. Cheeses were evaluated regarding their physicochemical composition, proteolysis, texture profile and viability of microorganisms after 1, 7, 14, 28, 40 and 60 days of refrigerated storage. Probiotics viability during simulated gastrointestinal conditions was evaluated after 28 and 60 days of storage. The results were evaluated using Analysis of Variance (ANOVA), and comparison of means by Tukey¿s test at 5% significance level. Descriptive sensory analysis was performed using free-choice profiling after 28 and 60 days of cheese storage, and the results were submitted to Generalized Procrustes Analysis (GPA). Both control and probiotic cheeses presented typical composition of Prato cheese and the treatments did not affect cheese composition. During 60 days of storage, an increased proteolysis, and decreased firmness was observed for all cheeses, and high populations of L. acidophilus and Bifidobacterium were found for the probiotic cheeses. Probiotic cheese showed similar sensory profiles with characteristics aroma and taste of Prato cheese more pronounced than control cheeses. The probiotics added to the cheeses were resistant during the simulation of the passage through the gastrointestinal tract and bile. In conclusion, the results evidenced that the addition of probiotics in Prato cheese proved to be a viable alternative, since the viability of microorganisms remained during cheese processing and storage, without causing undesirable changes in the sensory characteristics of the product, and provided protection to the probiotics during simulated gastrointestinal conditions / Doutorado / Tecnologia de Alimentos / Doutora em Tecnologia de Alimentos

O objetivo deste estudo foi o de avaliar a eficiência da aderência de produtos de exclusão competitiva e de probiótico sobre a tonsila cecal de frangos SPF, com idade de um a doze dias. Através da microscopia eletrônica de varredura, verificou-se a ação destes produtos comerciais com relação a proteção ao epitélio, quando desafiados com Salmonella Enteritidis. O delineamento experimental foi dividido em cinco (05) grupos, sendo o grupo T1 o controle negativo, o grupo T2 recebeu o produto de exclusão competitiva Aviguard, o grupo T3 recebeu o produto de exclusão competitiva Broilact, o grupo T4 recebeu o probiótico Biotop e o grupo T5 o controle positivo. Após 12 horas do tratamento com os produtos de exclusão competitiva ou probiótico as aves foram infectadas, por via oral, com S. Enteritidis. Os resultados obtidos mostraram que os produtos de exclusão competitiva e probiótico apresentaram uma maior eficiência em estabelecer uma microbiota protetora contra S. Enteritidis, e que a adição destes pode produzir um efeito de melhor índice zootécnico, quando comparado com as aves que foram desafiadas com S. Enteritidis e que apresentaram um ganho de peso inferior. / The aim of this study was to evaluate the ultra structure of the cecal tonsil in SPF chicks using scanning electron microscopy, determining the efficacy of competitive exclusion products and probiotic and its protection in cecal epithelium against an oral challenge with Salmonella Enteritidis. The experimental design included five groups: T1 negative control, T2 chicks treated with Aviguard, T3 chicks treated with Broilact, T4 chicks treated with Biotop, and, T5 positive control. After 12 hours of treatment with competitive exclusion products or probiotic the chicks were infected orally with S. Enteritidis. Results showed that treated groups had a protective effect stimulating the establishment of the resident microflora against pathogenic bacteria. Treated chicks showed high weight gain when compared to chicks challenged.

Les probiotiques sont des microorganismes vivants qui, ingérés en quantité suffisante, exercent des effets positifs sur la santé. La lyophilisation est aujourd’hui questionnée quant à sa consommation d’énergie et son caractère discontinu. S’il offre une alternative pour produire massivement des poudres probiotiques à faible coût, le séchage par atomisation induit quant à lui des stress thermiques et oxydatifs conduisant à des pertes de viabilité rédhibitoires.Dans ce travail, un procédé innovant de séchage par atomisation est proposé. Du lactosérum doux concentré (jusqu’à 30% p/p) est utilisé à la fois comme support de culture et de séchage de P. freudenreichii et L. casei. Ce procédé élimine les étapes intermédiaires à risque de contamination élevé, accroît la biomasse et améliore la viabilité des bactériesLes mécanismes sous-jacents ont été explorés au plan de la résistance bactérienne et des conditions de séchage. Le milieu concentré induit une osmoadaptation des bactéries par expression de protéines de stress et accumulation de solutés compatibles, conduisant à une tolérance accrue des probiotiques à différents stress. La présence d’agrégats et la concentration en Mg2+ du milieu concentré pourraient également être impliquées.Le scale-up du procédé a été étudié : un schéma technologique semi-industriel impliquant séchage par atomisation, sur bande et en lit fluidisé a permis d’atteindre une viabilité de 100% (> 109 CFU g-1). Par ailleurs, la fonctionnalité des poudres probiotiques a été évaluée in vitro and in vivo sur modèle porcelet. Ce travail ouvre de nouvell / Probiotics are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. Freeze drying, the reference drying method, is currently challenged because of its low energy-efficiency and productivity. Therefore, spray drying is expected to be an alternative and sustainable method for producing probiotic powders. The issue remains in the considerable inactivation of probiotics caused by high temperature and dehydration during the process. In this work, a novel spray-drying process for continuous production of probiotics was challenged. Concentrated sweet whey (up to 30% w/w dry matter) was used to both culture and spray dry P. freudenreichii ITG P20 and L. casei BL23. This process cut down the steps between culturing and drying (e.g. harvesting, washing), increases the cell population after growth and improves spray drying productivity and probiotic viability. The mechanisms were explored from bacterial physiology and drying process conditions. The hypertonic stress led to overexpression of key stress proteins and accumulation of intracellular compatible solutes, which enhanced multistress tolerance. The presence of protein aggregates and optimal concentration of Mg2+ in matrix may also be involved.The feasibility of scaling up this process was validated. A multi-stage semi industrial drying process, coupling spray-drying with belt drying and fluid-bed drying, was applied to further improve the probiotic viability to 100% (> 109 CFU g-1). Moreover, the functionality of these probiotic powders was investigated in vitro and in viv

As the demand for farmed shrimp continues to grow worldwide, the use of probiotics to address the sustainability of aquaculture fisheries has gained much attention. Emerging diseases in shrimp aquaculture, such as acute hepatopancreatic necrosis disease (AHPND), have devastating economic impacts in countries that largely depend on this activity. The relevance of this research lies on the fact that it explores the potential of using probiotics to mitigate the negative effects of AHPND in shrimp aquaculture. The scope of these studies includes survival of probiotic microbes in typical aquaculture water conditions, the effectiveness of probiotics in vitro and in vivo against the pathogenic strain of Vibrio parahaemolyticus that causes AHPND, and the effects of probiotics on the bacterial community composition in aquaculture water and gastrointestinal tract of shrimp after an induced AHPND infection. The microorganisms chosen as probiotics for this research include a lactic acid bacterium, a yeast and a photosynthetic bacterium. Informal feedback from shrimp farmers in Thailand and Vietnam revealed positive results against AHPND when using a commercially available probiotic containing multiple species of microorganisms from these probiotic groups. This research was divided into four studies. The first study (Chapter 2) evaluated the growth of the three different probiotic microbes in two different salinity conditions commonly found in intensive shrimp production systems to determine whether they could be further considered as potential candidates. The hypothesis was that the NaCl concentrations of the media may not have an effect on acid production, growth and cell morphology of the microorganisms being evaluated due to their metabolic mechanisms of adaptation to differences in osmotic pressure. The probiotic microbes were cultured in nutrient media enriched with 1 and 2% NaCl. Microbial survival, acidity and cell morphology between treatments were compared using enumeration by serial dilutions and plating, pH measurements and scanning electron microscopy imaging, respectively. The results showed that salinity levels up to 2% NaCl did not affect the growth of lactic acid bacteria and yeast. Photosynthetic bacteria grown in media with 1% NaCl showed a 24-hour delay in comparison to the control and a prolonged lag phase that lasted 48 hours when the media contained 2% NaCl. Therefore, the hypothesis was partially supported. Based on these results, all three probiotic microbes demonstrated to be suitable for application in aquaculture ponds with up to 2% salinity. The second study (chapter 3) aimed at determining the inhibitory effects of eight different formulations of probiotic solutions against the pathogenic strain of V. parahaemolyticus in vitro. The hypothesis of this study was that probiotic solutions containing whole microbial cultures of multiple microbial types including lactic acid bacteria may have a greater inactivation of the pathogen. The probiotic formulations consisted of individual cultures, combinations of the three probiotic microbes, and a commercially available probiotic formulation. The inhibitory effects were evaluated following a disk diffusion test on solid media by comparing diameters of zones of inhibition, and a challenge test in liquid media by comparing pathogen survival after exposure to probiotic solutions. Findings revealed inhibition zones with greater diameters in disks treated with whole microbial cultures (min: 7.83 mm, max: 11.33 mm) versus disks treated with only supernatants (min: 7.00 mm, max: 8.50 mm). Results from the challenge in liquid media tests showed greater inactivation of the pathogen after 48 h (6.56±0.07 to 5.43±0.03 log10 reduction) when treated with lactic acid bacteria alone and in combination with other microbial types. From these results, the hypothesis was supported and it was concluded that probiotic solutions including a lactic acid bacterium, the combination of lactic acid bacterium and photosynthetic bacterium and the combination of lactic acid bacterium, yeast and photosynthetic bacterium may be used to effectively inhibit AHPND in shrimp aquaculture. The third study (chapter 4) explored the effects of probiotic solutions on live shrimp (Litopenaeus vannamei) pretreated with probiotics for 7 days prior to challenging them with the pathogenic V. parahaemolyticus strain causing AHPND. The hypothesis of this study was that higher shrimp survival and weight gains would be observed when shrimps are exposed to probiotics solutions with multiple microbial types in the water and feed. Water quality parameters (dissolved oxygen, temperature, acidity, salinity and total ammonia nitrogen), difference in shrimp weight increase and shrimp survival were compared between probiotic treatments and controls. Treatments included: (1) a lactic acid bacterium alone (Pro.Sol1), (2) a lactic acid bacterium and a photosynthetic bacterium (Pro.Sol2), (3) the combination of a lactic acid bacterium, a yeast, and a photosynthetic bacterium (Pro.Sol3), and (4) a commercial probiotic (Com.Pro) and the results showed shrimp survival of 11.7, 26.7, 36.7 and 73.3%, respectively. Also, treatments Pro.Sol3 and Com.Pro resulted in higher weight gains (19.7 and 31.2%, respectively) versus the negative control (11.2%). Moreover, onset of the disease was delayed in all treatments as follows: 12 h with Pro.Sol1, 20 h with Pro.Sol2, 22 h with Pro.Sol3, and 26 h with Com.Pro. From these results, the hypothesis was supported and it was concluded that probiotics have the potential to effectively mitigate the effects of AHPND in the shrimp aquaculture. Finally, the fourth study (chapter 5) evaluated the effects of probiotics on the bacterial diversity of the gastrointestinal tract of shrimp as well as variation of bacterial and fungal diversity in the water before and after challenging shrimp with the pathogenic V. parahaemolyticus strain causing AHPND. The hypothesis of this study was that probiotic solutions with multiple microbial types may be able to maintain the microbial composition of the shrimp GI tract and aquaculture water preventing an increase in relative abundance of the family Vibrionaceae. Next generation sequencing was conducted using an Illumina MiSeq™ and primers specific for bacterial V4 hypervariable region of the 16S rRNA gene. The results obtained from the GI tract of shrimp revealed that the relative abundance of the family Vibrionaceae significantly increased in treatments with high mortalities, whereas treatments with higher survivals showed no significant difference in relative abundance of Vibrionaceae family members (P>0.05) in comparison to the negative control. The Shannon diversity index values (abundance and evenness) of the bacterial communities revealed that the treatment with the highest survival had the highest Shannon index value (4.69±0.133) whereas the treatment with lowest survival had the lowest Shannon index value (0.17±0.004). The results obtained from water samples did not show a higher abundance of the family Vibrionaceae, and diversity was maintained after infection (Shannon index 4.64±0.58). Regarding fungal diversity in water samples, Shannon index values revealed no significant changes before (3.627±0.37) and after infection (3.664±0.18) except for Pro.Sol3 (2.859±0.56) and Com.Pro (1.795±0.50), which included yeast in their formulation. Thus, the hypothesis of this study was partially supported since the results revealed that while all probiotics maintained the diversity of microbial composition in the water, only those probiotic solutions with various microbial types in the formulation maintained the diversity of the microbial composition in the GI tract of shrimp providing protection against AHPND.

Förekomsten av övervikt och fetma har ökat kraftigt de senaste åren över hela världen och i fetmans fotspår ökar även fetmarelaterade sjukdomar. Utvecklingen anses bero på en kombination av faktorer som större tillgång på energirik kost, miljömässiga, livsstilsrelaterade, genetiska och patologiska faktorer. Ny forskning har gett en ökad kuskap om tarmflorans betydelse för hälsa och studier på både obesa människor och djur visar att deras sammansättning i tarmfloran skiljer sig jämfört med normalviktiga individer. Detta har lett till ett ökat intresse hos forskare att titta närmare på kostens relation tilltarmfloran och dess sammansättning för att få klarhet i dess koppling till fetma och kunna använda denna kunskap för att förebygga och utveckla behandlingsmetoder mot fetma. Syftet med denna litteraturstudie var att analysera vetenskapliga artiklar och titta närmare på relationen mellan tarmfloran, kost och fetma. Resultaten visade att tarmflorans sammansättning är olika hos normalviktiga och obesa individer och att sammansättningen påverkar fermentationen av ej nedbrytningsbara kolhydrater i kolon. Studierna visade också att det finns en koppling mellan tarmfloran och inflammation som i sin tur också är en faktor relaterad till fetma. Ytterligare studier krävs för att besvara frågan ifall sammansättningen av tarmfloran är en orsakande faktor till utveckling av fetma, eller ett resultat av sjukdomen, samt vilka bakterier och grupper i den som genom bland annat kost går att påverka på ett positivt sätt mot fetma. / Obesity has increased dramatically during the past decades over the whole world, and has resulted in an increase of obesity-related diseases. The potential contributing factors to obesity are a combination of increase in the availability of energy-rich foods, environmental, lifestyle-related, genetic and pathological factors. New research has led to more knowledge about the gut microbiota and its role in health and studies show a difference in the microbial communities of lean vs. obese humans and animals. These findings have created an interest in research to understand gut microbiota composition and its relation to obesity so that the knowledge can be used in the prevention and treatment of obesity. The aim of this project was to analyse scentific articles and investigate the relation between the gut microbiota, diet and obesity. The studies showed differences in gut microbiota composition between lean vs. obese individuals, and that the composition affects the microbiotas ability to ferment non-digestible carbonhydrates in the colon. The studies also showed that the gut microbiota is related to inflammation, and inflammation is another factor that´s also related to obesity. There is a need for further studies to answer the question if the composition of the gut microbiota is the cause or the consequence of obesity, and which bacteria that for example through dietary modulation, can have a positive effect on obesity.

Les probiotiques sont des micro-organismes vivants, capables d’interagir avec le microbiote et les cellules de l’organisme hôte. Par leurs présences ou leurs métabolismes, ils contribuent notamment à la régulation du système immunitaire. D’un point de vue technologique, le développement d’un complément alimentaire contenant des probiotiques nécessite la maitrise des étapes de sélection et de production d’une bactérie fonctionnelle sous forme sèche et revivifiable. Au cours de ce travail, nous avons développé des tests de criblage en cytométrie en flux, permettant de déterminer l’aptitude d’une bactérie à être séchée et l’origine de sa sensibilité. Pour mettre en place ces tests, nous nous sommes intéressés aux mécanismes mis en jeu lors du procédé de séchage. Ainsi, lors de la déshydratation, les micro-organismes vont subir simultanément un stress osmotique et un stress oxydant. Les résultats montrent un fort impact du séchage sur la viabilité et le potentiel immuno-modulateur avec une diminution de la composante pro-inflammatoire (IL-12). Pour protéger les bactéries, trois stratégies d’optimisation ont été développées : le séchage par formation de mousse, l’encapsulation dans une matrice alginate-protéines de pois, et le co-séchage d’une bactérie fragile avec une bactérie résistante. Les connaissances acquises au cours de cette étude ont permis d’optimiser la production industrielle de la souche finale du projet. Afin de répondre au nouveau cadre réglementaire européen, une étude clinique en double aveugle contre placébo est actuellement en cours (2011-2013) / Probiotics are live microorganisms that can act on the immune system at specific sites in the gastro-intestinal tract. From a technological point of view, the development of a dietary supplement containing viable and functional probiotics requires the control of the selection and production steps. In a first time, we have developed specific and sensitive screening tests to determine the ability of bacteria to be dried and the origin of its sensitivity. To develop these tests, we investigated the mechanisms involved during drying process. Thus, microorganisms simultaneously undergo osmotic stress and oxidative stress. Results show a strong impact of drying on the survival rate and immunomodulatory potential with a decrease of proinflammatory cytokines (IL-12). To protect probiotic bacteria, three optimization strategies have been developed: drying by foam formation, encapsulation of probiotic bacteria using a pea-protein alginate matrix and drying of a sensitive probiotic strain with a resistant bacterium. Knowledge obtained during this study was used to optimize the industrial process of the final strain of the project. A randomized double blinded (RDB) trial is in progress (2011-2013) to test the bacteria in humans