Uma notável tendência na área de alimentos é o desenvolvimento de produtos que agreguem sabor, praticidade, conveniência e tragam benefícios à saúde. Dentro desse cenário, encontram-se os alimentos funcionais, que têm sido alvos de pesquisas científicas para elucidar os benefícios que trazem ao homem. Os probióticos e prebióticos são os ingredientes fisiologicamente ativos que compõem o alimento e agem na manutenção do equilíbrio da microbiota do trato gastrintestinal humano. A combinação de probióticos e prebióticos resulta em um produto simbiótico. Com outro apelo de alimento funcional, pode-se citar o tomate, fruto que apresenta licopeno em sua composição, pigmento com potencial antioxidante e ação preventiva contra diversos tipos de câncer. O presente trabalho teve como objetivo desenvolver um produto tipo patê potencialmente simbiótico, à base de queijo quark, adicionado de derivados de tomate, manjericão e queijo parmesão, bem como avaliar as suas características físico-químicas, microbiológicas, de textura instrumental e sua aceitabilidade sob o ponto de vista sensorial durante o seu armazenamento a 4 ± 1 °C por até 28 dias. Foram produzidos três tratamentos do produto tipo patê (em triplicata), todos eles suplementados com inulina: T1 (controle) com a cultura starter Streptococcus thermophilus ST-M6; T2 com a cultura starter ST-M6 e a adição dos probióticos Bifidobacterium animalis subsp. lactis Bb-12 e Lactobacillus acidophilus NFCMe T3 com a cultura starterST-M6, a cultura probiótica Bb-12 e a adição de Lactobacillus sakei 2a. Os produtos foram armazenados a 4 ± 1 °C e durante os períodos de amo stragem (1, 7, 14, 21 e 28 dias), foi analisada a viabilidade dos probióticos, da cultura starter e de L. sakei 2a, a fim de avaliar a possível adaptação desta cepa bacteriocinogênica ao meio lácteo, uma vez que foi isolada de linguiça frescal. Adicionalmente, o pH e o perfil de textura instrumental do produto (teste de dupla penetração de amostras em analisador de textura TA-XT2) foram analisados e, a partir de amostras mantidas congeladas no dia seguinte à sua fabricação, foi determinada a composição centesimal das diferentes formulações. Além disso, aos 7, 14 e 21 dias, as amostras foram avaliadas sensorialmente (teste de aceitabilidade com escala hedônica estruturada de 9 pontos) por 50 provadores em cada seção. Paralelamente, foi feito um monitoramento microbiológico das amostras quanto à presença de contaminantes. O pH dos três tratamentos apresentou uma queda significativa ao longo dos 28 dias (0,16 a 0,21) e foi significativamente menor (p<0,05) para T2 (4,67 após 28 dias) em relação a T1 (4,85) e T3 (4,76). Quanto à viabilidade da cultura starter, os produtos apresentaram populações médias ao longo do armazenamento sempre acima de 8,95 log UFC/g. As culturas probióticas NCFM e Bb-12 apresentaram contagens acima do mínimo recomendado pela legislação durante o armazenamento, com populações acima de 8 log UFC/g para Bb- 12 e populações de NCFM que diminuíram significativamente (p<0,05) de 8,61 (7 dias) a 7,93 log UFC/g (28 dias) para T2. L. sakei 2a mostrou-se adaptar bem ao produto lácteo, com viabilidade que se manteve na faixa de 8,10 a 8,18 log UFC/g ao longo dos 28 dias. Quanto ao perfil de textura, T2 foi mais estável durante o armazenamento, com parâmetros mais constantes ao longo do estudo, em comparação a T1 e T3. Na análise sensorial, não foram encontradas diferenças significativas (p>0,05) entre os tratamentos durante os dias avaliados e, tampouco, entre os dias estudados para um mesmo tratamento. As amostras tiveram uma boa aceitação pelos provadores, principalmente T2, com notas médias acima de 7 (gostei regularmente), apesar de não diferirem significativamente. O produto inovador tipo patê de base láctea no sabor tomate desenvolvido mostrou-se um bom veículo para os micro-organismos probióticos B. animalis Bb-12 e L. acidophilus NCFM e para L. sakei 2a, potencialmente simbiótico e sensorialmente aceitável. / A remarkable trend in the food area is the development of products that are tasty, convenient, and able to bring healthy benefits. Within this scenario, functional foodshave been object of scientific studies to elucidate their benefits to the human being. Probiotics and prebiotics are physiologically active ingredients in food which positively influence the maintenance of the human gastrointestinal tract balance. A synbiotic product combines probiotic and prebiotics ingredients. The tomato is also remarkable, regarding some compounds with functional attributes in its composition. Lycopene is the pigment present in the fruit that has antioxidant potential and is correlated with reduced incidence of some cancer types. This study aimed to develop a potentially synbiotic quark cheese-based spread product containing tomato and with basil and parmesan cheese, as well as evaluating its physico-chemical and microbiological features, and its instrumental texture profile and sensory acceptability, during storage at 4 ± 1 °C f or up to 28 days. Three different trials of quark cheese-based spread product were produced (in triplicates), all supplemented with inulin: T1 (control) with the starter culture Streptococcus thermophilus ST-M6; T2 with the starter culture ST-M6 plus the addition of the probiotics Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp. lactis Bb-12, and T3 with the starter culture ST-M6, the probiotic culture Bb-12, and the addition of Lactobacillus sakei 2a. The products were stored at 4 ± 1 °C and, duri ng the sampling periods (1, 7, 14, 21, and 28 days), the viability of probiotics, starter culture, and L. sakei 2a was assessed. The adaptation of L. sakei 2a to the dairy matrix was also evaluated, once this bacteriocin-producing strain was isolated from a Brazilian sausage. Additionally, the pH and instrumental texture profile of the product was assessed (double penetration of samples test, using a TA-XT2 texture analyzer), and the chemical composition was determined, using samples kept frozen after the first day of production. Besides, the trials were sensory evaluated (acceptability test, using a 9-point structured hedonic scale), after 7, 14, and 21 days, by 50 consumers in each session. At the same time, samples were microbiologically monitored for the presence of contaminants. The pH of the three trials displayed a significantly decrease throughout the 28 days of storage (0.16 up to 0.21), and it was significantly lower (p<0.05) for T2 (4.67 after 28 days), compared to T1 (4.85) and T3 (4.76). Regarding the viability of the starter culture, the products showed mean populations always above 8.95 log CFU/g during storage. The probiotic cultures NCFM and Bb-12 displayed counts above the minimum recommended by the Brazilian regulatory standards throughout storage, with populations above 8 log CFU/g for Bb-12, and counts of NCFM decreased significantly (p<0.05) from 8.61 (7 days) to 7.93 log CFU/g (28 days) for T2. L. sakei 2a was able to adequately adapt to the dairy product, with populations that maintained between 8.10 and 8.18 log CFU/g during the 28 days of storage. With respect to the texture profile, T2 was considered the most stable trial during storage, as displayed preservation ofmore homogeneous parameters throughout study, compared to T1 and T3. Regarding sensory evaluation, there were neither significantly differences (p>0,05) among the trials during the studied period nor among the studied periods for the same trial. The samples had a good acceptance by consumers, with mean scores above 7 (like regularly), mainly T2, although the trials did not differ significantly. The innovative tomato dairy-based spread product developed revealed to be an adequate matrix for the probiotic microorganisms B. animalis Bb-12 and L. acidophilus NCFM, and for L. sakei 2a, potentially synbiotic, and sensory acceptable.

Dans les produits laitiers, les cultures bioprotectrices et leurs métabolites représentent une alternative d’intérêt aux conservateurs chimiques pour lutter contre les contaminations fongiques. L’objectif de cette thèse était de développer des ingrédients antifongiques, issus de la fermentation d’un substrat laitier par différents microorganismes, bactéries ou champignons, utilisables dans des produits laitiers variés. Pour ce faire, nous avons dans un premier temps caractérisé la diversité des contaminants fongiques des produits laitiers et de leur environnement afin de sélectionner les cibles fongiques les plus pertinentes pour le reste de notre étude. Ensuite, nous avons criblé, in vitro, l’activité antifongique de fermentats issus de la fermentation de 2 substrats laitiers par 698 souches de bactéries lactiques, propioniques et de champignons grâce à une nouvelle méthode de criblage haut-débit, dans une matrice mimant le fromage. Après optimisation des conditions de fermentation pour améliorer l’activité antifongique de ces fermentats, les plus actifs ont été testés à l’échelle du laboratoire sur des fromages blancs et des fromages à pâte pressée à croûte morgée (PPCM) avant d’être testés à l’échelle pilote dans des crèmes fraiches et des fromages à PPCM. Cette étape nous a permis (i) de valider l’activité antifongique des fermentats en produits réels en mettant en place des challengetests et des tests d’usage et (ii) d’évaluer leur impact sur les qualités organoleptiques des produits grâce à des analyses sensorielles. Les molécules impliquées dans l’activité antifongique ont ensuite été identifiées grâce à différentes méthodes (chromatographie en phase gazeuse ou liquide couplée ou non à la spectrométrie de masse) et l’impact de ces composés sur la croissance des cibles fongiques a été évalué. Le criblage in vitro a permis la sélection de 3 ingrédients antifongiques qui se sont tous révélés actifs in situ. Les tests de durabilité ont révélé que le fermentat issu de Lactobacillus rhamnosus CIRM-BIA 1952 avait une activité prometteuse dans les crèmes fraiches. Pour le fromage à PPCM, le fermentat issu de Propionibacterium jensenii CIRM-BIA 1774, qui retarde jusqu’à 16 jours la croissance de Mucor racemosus et Penicillium commune pourrait être utilisé à la place de la natamycine. La caractérisation des composés antifongiques dans les fermentats a mis en évidence certains acides organiques, des acides gras, des composés volatils et un peptide qui, ensemble, jouent très probablement un rôle dans l’activité antifongique de ces fermentats. Nous avons enfin montré l’effet fongistatique du fermentat le plus actif sur Rhodotorula mucilaginosa UBOCC-A-216004 et Mucor racemosus UBOCC-A-109155 ainsi que son impact sur la physiologie des spores de M. racemosus. L’ensemble de ces résultats devrait donc conduire au développement de nouveaux ingrédients laitiers antifongiques qui pourraient remplacer avantageusement les conservateurs dans les produits laitiers, ils apportent en outre des éléments d’information sur les molécules et les mécanismes mis en œuvre dans l’inhibition antifongique. / In dairy products, bioprotective cultures and their metabolites represent an interesting alternative to chemical preservatives. The aim of this PhD thesis was to develop antifungal dairy ingredients, derived from the fermentation of dairy substrates by different microorganisms, bacteria and fungi, which could be used in various dairy products. To do so, we first studied the diversity of spoilage fungi in a large variety of dairy products and their environment in order to select the most appropriate fungal targets. Then, we screened the in vitro antifungal activity of fermentates obtained from the fermentation of 2 dairy substrates by 698 lactic acid bacteria, propionibacteria and fungal strains using a novel high-throughput method in a cheese mimicking model. After optimizing fermentation conditions to enhance antifungal activity, the most active fermentates were tested at a lab scale in semi-hard and fresh cheese before evaluation at a pilot scale in semi-hard cheese and sour cream. This step allowed us (i) to validate the fermentate antifungal activities in real products using challenge- and durability-tests and (ii) to evaluate their impact on the products’ organoleptic properties using sensorial analysis. Antifungal molecules were then identified using different analytical methods (high performance liquid chromatography, gas chromatography coupled or not with mass spectrometry) and impact of these compounds on fungal growth was studied. The in vitro screening allowed selecting 3 antifungal ingredients that were also active in situ. Durability tests revealed that Lactobacillus rhamnosus CIRMBIA 1952 fermentate had a promising activity in sour cream. In semi-hard cheese, Propionibacterium jensenii CIRMBIA 1774 fermentate, which delayed Mucor racemosus and Penicillium commune growth for up to 16 days, could be used instead of natamycin. Antifungal compounds present in fermentates consisted of organic acids, free fatty acids, volatile compounds and peptides which altogether might play a role in the antifungal activity of these fermentates. Finally, the fungistatic effect of P. jensenii CIRM-BIA 1774 fermentate against R. mucilaginosa UBOCC-A-216004 and M. racemosus UBOC-A-109155 was demonstrated, as well as its impact on the physiology of M. racemosus spores. Together, these results should lead to the development of new antifungal dairy ingredients which could replace preservatives in dairy products. Finally, these results gave us new information concerning antifungal molecules and their action mechanisms.

Uma notável tendência na área de alimentos é o desenvolvimento de produtos que agreguem sabor, praticidade, conveniência e tragam benefícios à saúde. Dentro desse cenário, encontram-se os alimentos funcionais, que têm sido alvos de pesquisas científicas para elucidar os benefícios que trazem ao homem. Os probióticos e prebióticos são os ingredientes fisiologicamente ativos que compõem o alimento e agem na manutenção do equilíbrio da microbiota do trato gastrintestinal humano. A combinação de probióticos e prebióticos resulta em um produto simbiótico. Com outro apelo de alimento funcional, pode-se citar o tomate, fruto que apresenta licopeno em sua composição, pigmento com potencial antioxidante e ação preventiva contra diversos tipos de câncer. O presente trabalho teve como objetivo desenvolver um produto tipo patê potencialmente simbiótico, à base de queijo quark, adicionado de derivados de tomate, manjericão e queijo parmesão, bem como avaliar as suas características físico-químicas, microbiológicas, de textura instrumental e sua aceitabilidade sob o ponto de vista sensorial durante o seu armazenamento a 4 ± 1 °C por até 28 dias. Foram produzidos três tratamentos do produto tipo patê (em triplicata), todos eles suplementados com inulina: T1 (controle) com a cultura starter Streptococcus thermophilus ST-M6; T2 com a cultura starter ST-M6 e a adição dos probióticos Bifidobacterium animalis subsp. lactis Bb-12 e Lactobacillus acidophilus NFCMe T3 com a cultura starterST-M6, a cultura probiótica Bb-12 e a adição de Lactobacillus sakei 2a. Os produtos foram armazenados a 4 ± 1 °C e durante os períodos de amo stragem (1, 7, 14, 21 e 28 dias), foi analisada a viabilidade dos probióticos, da cultura starter e de L. sakei 2a, a fim de avaliar a possível adaptação desta cepa bacteriocinogênica ao meio lácteo, uma vez que foi isolada de linguiça frescal. Adicionalmente, o pH e o perfil de textura instrumental do produto (teste de dupla penetração de amostras em analisador de textura TA-XT2) foram analisados e, a partir de amostras mantidas congeladas no dia seguinte à sua fabricação, foi determinada a composição centesimal das diferentes formulações. Além disso, aos 7, 14 e 21 dias, as amostras foram avaliadas sensorialmente (teste de aceitabilidade com escala hedônica estruturada de 9 pontos) por 50 provadores em cada seção. Paralelamente, foi feito um monitoramento microbiológico das amostras quanto à presença de contaminantes. O pH dos três tratamentos apresentou uma queda significativa ao longo dos 28 dias (0,16 a 0,21) e foi significativamente menor (p<0,05) para T2 (4,67 após 28 dias) em relação a T1 (4,85) e T3 (4,76). Quanto à viabilidade da cultura starter, os produtos apresentaram populações médias ao longo do armazenamento sempre acima de 8,95 log UFC/g. As culturas probióticas NCFM e Bb-12 apresentaram contagens acima do mínimo recomendado pela legislação durante o armazenamento, com populações acima de 8 log UFC/g para Bb- 12 e populações de NCFM que diminuíram significativamente (p<0,05) de 8,61 (7 dias) a 7,93 log UFC/g (28 dias) para T2. L. sakei 2a mostrou-se adaptar bem ao produto lácteo, com viabilidade que se manteve na faixa de 8,10 a 8,18 log UFC/g ao longo dos 28 dias. Quanto ao perfil de textura, T2 foi mais estável durante o armazenamento, com parâmetros mais constantes ao longo do estudo, em comparação a T1 e T3. Na análise sensorial, não foram encontradas diferenças significativas (p>0,05) entre os tratamentos durante os dias avaliados e, tampouco, entre os dias estudados para um mesmo tratamento. As amostras tiveram uma boa aceitação pelos provadores, principalmente T2, com notas médias acima de 7 (gostei regularmente), apesar de não diferirem significativamente. O produto inovador tipo patê de base láctea no sabor tomate desenvolvido mostrou-se um bom veículo para os micro-organismos probióticos B. animalis Bb-12 e L. acidophilus NCFM e para L. sakei 2a, potencialmente simbiótico e sensorialmente aceitável. / A remarkable trend in the food area is the development of products that are tasty, convenient, and able to bring healthy benefits. Within this scenario, functional foodshave been object of scientific studies to elucidate their benefits to the human being. Probiotics and prebiotics are physiologically active ingredients in food which positively influence the maintenance of the human gastrointestinal tract balance. A synbiotic product combines probiotic and prebiotics ingredients. The tomato is also remarkable, regarding some compounds with functional attributes in its composition. Lycopene is the pigment present in the fruit that has antioxidant potential and is correlated with reduced incidence of some cancer types. This study aimed to develop a potentially synbiotic quark cheese-based spread product containing tomato and with basil and parmesan cheese, as well as evaluating its physico-chemical and microbiological features, and its instrumental texture profile and sensory acceptability, during storage at 4 ± 1 °C f or up to 28 days. Three different trials of quark cheese-based spread product were produced (in triplicates), all supplemented with inulin: T1 (control) with the starter culture Streptococcus thermophilus ST-M6; T2 with the starter culture ST-M6 plus the addition of the probiotics Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp. lactis Bb-12, and T3 with the starter culture ST-M6, the probiotic culture Bb-12, and the addition of Lactobacillus sakei 2a. The products were stored at 4 ± 1 °C and, duri ng the sampling periods (1, 7, 14, 21, and 28 days), the viability of probiotics, starter culture, and L. sakei 2a was assessed. The adaptation of L. sakei 2a to the dairy matrix was also evaluated, once this bacteriocin-producing strain was isolated from a Brazilian sausage. Additionally, the pH and instrumental texture profile of the product was assessed (double penetration of samples test, using a TA-XT2 texture analyzer), and the chemical composition was determined, using samples kept frozen after the first day of production. Besides, the trials were sensory evaluated (acceptability test, using a 9-point structured hedonic scale), after 7, 14, and 21 days, by 50 consumers in each session. At the same time, samples were microbiologically monitored for the presence of contaminants. The pH of the three trials displayed a significantly decrease throughout the 28 days of storage (0.16 up to 0.21), and it was significantly lower (p<0.05) for T2 (4.67 after 28 days), compared to T1 (4.85) and T3 (4.76). Regarding the viability of the starter culture, the products showed mean populations always above 8.95 log CFU/g during storage. The probiotic cultures NCFM and Bb-12 displayed counts above the minimum recommended by the Brazilian regulatory standards throughout storage, with populations above 8 log CFU/g for Bb-12, and counts of NCFM decreased significantly (p<0.05) from 8.61 (7 days) to 7.93 log CFU/g (28 days) for T2. L. sakei 2a was able to adequately adapt to the dairy product, with populations that maintained between 8.10 and 8.18 log CFU/g during the 28 days of storage. With respect to the texture profile, T2 was considered the most stable trial during storage, as displayed preservation ofmore homogeneous parameters throughout study, compared to T1 and T3. Regarding sensory evaluation, there were neither significantly differences (p>0,05) among the trials during the studied period nor among the studied periods for the same trial. The samples had a good acceptance by consumers, with mean scores above 7 (like regularly), mainly T2, although the trials did not differ significantly. The innovative tomato dairy-based spread product developed revealed to be an adequate matrix for the probiotic microorganisms B. animalis Bb-12 and L. acidophilus NCFM, and for L. sakei 2a, potentially synbiotic, and sensory acceptable.

A dairy-based whipped topping consisting of 22.0 percent milk fat, 7.5 percent milk solids-not-fat, 12.0 percent sucrose, 10.0 percent corn syrup solids, 0.60 percent gum arabic, 0.06 percent carrageenin, 0.19 percent polyoxyethylene sorbitan rnonostearate, 0.19 percent polyoxythylene sorbitan tristearate, and 0.12 percent sodium stearoyl-2-lactylate was developed that would withstand the rigors of frozen storage. The best products were obtained when the topping was whipped on a Creamery Package 3M-30 continuous type ice cream freezer. Toppings whipped in the laboratory at temperatures higher or lower than -2.2 to -1.0 centigrade were weak and slightly wet. After 18 days frozen storage the toppings whipped at -2.2 centigrade had the best body and texture characteristics. Refrigerated storage after 18 days frozen storage resulted in an enlargement of the air cells and after about 10 days a stale flavor was detected. Commercial application of the formulation was considered feasible.