Dissertations / Theses on the topic 'Probiotic fermented milk'

The aim of this research was to study the identity and survival of probiotic bifidobacteria in fermented dairy products from the UK market throughout their shelf- life and carry out in vitro studies on some of the probiotic criteria and potential functional properties of the isolates including resistance to stomach-intestine conditions, antimicrobial activities, antibiotic resistance/susceptibility, antibiotic resistance genes and their transferability, biofilm formation and production of health- promoting secondary metabolites, such as conjugated linoleic acid (CLA). The initial study for choosing the right selective medium for isolation and enumeration of bifidobacteria included a comparative study on selectivity of well known media transgalactosylated oligosaccharides-mupirocin lithium salt (TOS- MUP) and Bifidobacterium iodoacetate medium 25 (BIM-25). Considering selectivity of the medium, recovery of the bifidobacteria and ease of preparation, it was concluded that TOS-MUP is the medium of choice for determination of total viable counts of bifidobacteria in fermented milks. The result of enumeration study indicated that most of the tested products (22 out of total 24) on TOS-MUP comply with the recommended minimum therapeutic level for probiotics (106 CFU/g) at the time of purchase and at the end of their shelf-life. All presumptive Bifidobacterium isolates were identified as Bifidobacterium animalis subsp. lactis by analysis of partial sequences of the 16S ribisomal RNA gene. Further use of rep-PCR for differentiation of all isolates using two different primers (GTG-5 and BOXA 1 R) did not show any difference among tested isolates as all isolates presented the same band profile. As far as the claims on the label are concerned, most of the products (19 out of 24) failed to name the Bifidobacterium species at all or correctly.

Orientador: Salvador Massaguer Roig
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Doutorado
Doutor em Tecnologia de Alimentos

A crescente preocupação com tecnologias sustentáveis e a procura de novos alimentos funcionais despertam o interesse para o desenvolvimento de novos produtos alimentícios que proporcionem, além da nutrição, benefícios à saúde do consumidor. Assim, esse trabalho visa propor o leite orgânico como potencial matéria-prima para a fabricação de leites fermentados probióticos. Para tanto, estudou-se o perfil tecnológico de cepas de bifidobactéria em cultura pura e em co-cultura com Streptococcus thermophilus em leites orgânico e convencional, analisando a composição química dos leites, determinando o perfil de acidificação de quatro cepas de Bifidobacterium animalis subsp. lactis em cultura pura e em co-cultura com Streptococcus thermophilus, verificando a contagem microbiológica das culturas probióticas e iniciadoras nos leites fermentados, examinando o perfil de ácidos graxos e o teor de ácido linoléico conjugado dos leites fermentados e determinando o perfil de textura dos leites fermentados. A maior velocidade de acidificação foi observada para as cepas B94 e BL04 em leite orgânico e para a cepa HN019, para ambos os tipos de leite. As contagens de todas as cepas de B. animalis subsp. lactis foram superiores a 8,58 log10 unidades formadoras de colônia (UFC).mL-1The concern about sustainable technologies and demand for new functional foods arouses the interest for the development of new food products in addition to provide nutrition and health benefits to the consumer. Thus, this work aims to offer organic milk as a potential raw material for the manufacture of probiotic fermented milk. Therefore, studying the technological profile of strains of bifidobacteria in pure culture and in co-culture with Streptococcus thermophilus in organic and conventional milks, analyzing the chemical composition of milk; determining the profile of acidification of four strains of Bifidobacterium animalis subsp. lactis in pure culture and in co-culture with Streptococcus thermophilus; checking the counts of starter and probiotic cultures in fermented milks; examining the profile of fatty acids and conjugated linoleic acid content of fermented milks and the profile of texture of fermented milks. The highest rate of acidification profile was observed for the strains BL04 and B94 in organic milk and for the strain HN019 in both milks. The counts of all strains of B. animalis subsp. lactis were higher than 8.58 log10 colony forming units (CFU). mL-1. The organic milk had higher levels of iron and protein, whereas conventional milk had higher levels of fat and lactose. The main fatty acids were not influenced by the type of milk and higher amounts of conjugated linoleic acid (65% higher than the control) were found in organic fermented milk with the strain BB12 in co-culture with S. thermophilus. Thus, it was found that organic milk can be used as raw material in the manufacture of probiotic fermented milk, increasing nutritional quality to final product.

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A ingestão de probióticos tem sido relatada como sendo um dos métodos mais amplamente utilizados para modular a microbiota intestinal. Curiosamente, o diabetes mellitus tipo 2 tem sido associado à disbiose e uma das possíveis vias de reestabelecer a microbiota intestinal saudável é por meio da ingestão regular de probióticos, os quais vem se destacando na indústria alimentícia. Assim, o objetivo deste estudo foi, primeiramente, desenvolver um leite fermentado de origem caprina, saborizado com suco de uva, contendo probióticos e, posteriormente, avaliar o seu efeito metabólico em diabéticos. Leites fermentados contendo ou não bactérias probióticas (Lactobacillus acidophilus La-5 e Bifidobacterium animalis subsp. lactis BB-12) foram submetidos a analises físico-químicas, microbiológicas, sensoriais, além da caracterização nutricional do produto, incluindo atividade antioxidante. O teor de compostos fenólicos totais e atividade antioxidante do leite fermentado contendo probióticos foi maior (p < 0,01) do que o leite fermentado convencional. Observou-se uma maior perda da viabilidade celular para L. acidophilus do que para o B. animalis. No entanto, a viabilidade de todas as bactérias foi adequada (> 106 UFC/ mL) até o 28o dia de armazenamento a 4 oC. Ambos os leites fermentados analisados apresentaram boas características sensoriais, não havendo diferença (p > 0,05) entre os mesmos. Um estudo duplo-cego, randomizado e placebo-controlado, incluindo 50 indivíduos diabéticos, foi desenvolvido posteriormente. Os diabéticos foram divididos em dois grupos, recebendo 120 mL/dia de uma das bebidas durante 6 semanas. Medidas antropométricas, de composição corporal, coleta de sangue e amostras fecais foram obtidos no início e ao final do estudo. A ingestão de leite fermentado contendo probióticos promoveu uma redução (p ≤ 0,05) nos níveis de frutosamina e uma tendência à redução (p = 0,07) nos níveis de hemoglobina glicada. Em ambos os grupos foram observados reduções significativas nos níveis de TNF-α e resistina e a concentração fecal de ácido acético aumentou ao final do estudo, enquanto os níveis de IL-10 foi reduzida (p < 0,001) apenas no grupo controle. Houve diferença significativa entre os grupos em relação às alterações de HbA1c, colesterol total e lipoproteína de baixa densidade. Não houve alterações (p > 0,05) na capacidade antioxidante total e F2-isoprostano. Este estudo desenvolveu uma bebida funcional com boa qualidade em termos de sobrevivência de bactérias e características sensoriais e nutricionais. A ingestão regular da bebida contendo probióticos melhorou o controle glicêmico em diabéticos, no entanto, a ingestão de leite fermentado caprino saborizado com suco de uva, esteve envolvido com outras alterações metabólicas.
The administration of probiotics and prebiotics has been reported to be one of the most widely used approaches to modulate intestinal microbiota. Interestingly, type 2 diabetes has been associated with dysbiosis and one of the possible routes for restore a healthy gut microbiota is by the regular ingestion of probiotics, which has been highlighted in the food industry. The present study aimed, first, to develop a flavored fermented goat milk containing probiotics and assess their metabolic effect in diabetics. Fermented milk with or without probiotic bacteria (Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12) were subjected to physicochemical, microbiological and sensory analysis, besides the nutritional characterization of the product, including antioxidant activity. Total phenolic contents and antioxidant activity of probiotic flavored fermented milk were significantly higher (p < 0.01) than conventional fermented milk. A higher loss in cell viability was observed for L. acidophilus than for the B. animalis. However, the viability of all bacteria was adequate (> 106 cfu/mL) until day 28 of storage. The fermented milk showed good sensory characteristics and no significant sensory preference among the fermented milks was found. A double-blind, randomized, placebo-controlled study including 50 diabetic patients, was developed later. The subjects were divided into two groups, receiving 120 mL/ day one of the fermented milks for 6 weeks. Anthropometric measurements, body composition, fasting blood and faecal samples were taken at baseline and after 6 weeks of intervention. The study demonstrated a significant decrease in fructosamine levels (p ≤ 0.05) and haemoglobin A1c tended to be lower (p = 0.07) in probiotic group. TNF- α and resistin were significantly reduced and faecal acetic acid was increased in both groups after the end of trial, while IL-10 was reduced (p < 0.001) only in the control group. There was a significant difference between groups concerning mean changes of HbA1c, total cholesterol and low-density lipoprotein. No significant changes (p > 0.05) from baseline were detected in plasma total antioxidant status and F2-isoprostane. This study developed a beverage of good quality, in terms of survival of bacteria and sensory and nutritional characteristics. Probiotic flavored fermented milk consumption improved the glycemic control in diabetic subjetcs, however, the intake of flavored fermented goat milk was involved with others metabolic changes.

Thesis presented in partial fulfilment of the requirements for the degree of Master of Technology (Food Technology) Department of Food Technology Faculty of Applied Sciences Cape Peninsula University of Technology, 2012
The aim of this study was to evaluate bambara groundnut milk (BGNM) subjected to fermentation with lactic acid bacteria (LAB) as a probiotic beverage with a view to developing value-added product. Central Composite Rotatable Design (CCRD) was used to optimise the hydration time and temperature of BGN flour for optimum BGN milk (BGNM) production. The optimum time and temperature was 2 h at 25oC. The effect of variety was assessed on the quality and consumer acceptability of BGNM prepared from five varieties of BGN (black, red, brown, brown-eye, and black-eye) which were representatives of the BGN available in South Africa. BGNM from the five varieties differed significantly (p<0.05) in, lightness, chroma, redness, yellowness, hue and antioxidative activity, while the pH were not significantly different. The four BGNM samples were significantly different (p < 0.05) in appearance, colour, mouthfeel and overall acceptability but not in aroma and taste. A three factor design (4 x 3 x 3) consisting of probiotics (Lactobacillus acidophilus, L. bulgaricus, L. casei and L. plantarum), temperature and fermentation time, were used to estimate the optimal conditions for the production of BGN probiotic beverage (BGNPB). The optimal condition for the production of BGNPB was estimated to be 35oC for 24 h with a desirability of 0.854 for L. bulgaricus. The next promising probiotic was L. plantarum that could be fermented at 35oC for 24 h with 0.843 desirability. BGNM from the red variety were fermented with L. bulgaricus and L. plantarum and L bulgaricus (in combination), making plain and sweetened BGNPB which were evaluated for their quality and consumer acceptability. The four BGNPB samples were significantly different (p < 0.05) in aroma, taste, mouthfeel and overall acceptability but not in appearance and colour. The plain BGNPB were assessed for their proximate composition, antioxidant activity, in vitro probiotic tolerance to simulated gastric juices and bile and a 28 days shelf life study at 5, 15 and 25oC. The protein, total dietary fibre (TDF), ash and antioxidative activity of the BGNPB were significantly different while the fat and carbohydrates were not significantly different. Time and concentration of the gastric juice and bile had significant effects on the percentage bacterial survival of probiotics in the BGNPB. However, the probiotics did survive, in low numbers, in the simulated gastric juice and bile after 180 and 240 minutes of incubation. Titratable acidity, pH, microbial load and colour of the BGNPB were significantly affected by the storage time and temperature during the shelf life study. At the 5oC storage temperature the BGNPB had a right censored shelf life on day 28. At 15oC the shelf life was 18 and 10 days for L bulgaricus and L. plantarum and L. bulgaricus respectively. The outcome of this research showed that a novel BGNPB product can be made from fermenting BGNM with LAB.

O trato gastrointestinal de humanos é rico em microrganismos que, podem tanto ser benéficos para a saúde do hospedeiro, prevenindo e/ ou tratando a intolerância à lactose, constipação intestinal, síndrome do intestino irritável, entre outras, quanto podem prejudicá-lo, afetando a aquisição de nutrientes e produção de mediadores inflamatórios. Estes distintos papéis da microbiota intestinal são tão marcantes que podem, inclusive, influenciar no desenvolvimento da obesidade em algumas pessoas, podendo levar até mesmo à Síndrome Metabólica. Probióticos e prebióticos podem conferir alterações nas propriedades da microbiota, afetando o crescimento bacteriano e seu metabolismo e, até mesmo, o uso de nutrientes. Assim, o objetivo geral deste trabalho foi avaliar o efeito do uso de leite fermentado simbiótico na obesidade e nos indicadores de Síndrome Metabólica, como circunferência de cintura, TGL, HDL-c, glicemia e pressão arterial, em pacientes com predisposição a esta doença, selecionados de acordo com os critérios de diagnóstico para síndrome metabólica. Para esta pesquisa foram desenhados quatro produtos, três deles inoculados com uma cultura comercial de probiótico - Bifidobacterium lactis (BL420), adicionados ou não de casca de maracujá em pó (prebiótico), sendo um deles preparado com uma emulsão de óleos vegetais, que apresenta efeito em prolongar a saciedade; e o quarto produto, fermentado apenas com a cultura clássica Streptococcus thermophilus. Avaliaram-se as propriedades tecnológicas dos produtos - físico-química, sensorial, microestrutura, microbiológica, e estudaram-se seus aspectos funcionais. Finalmente, os produtos foram testados em consumidores ao longo de um ensaio clínico durante 12 semanas, período no qual os voluntários tiveram que consumir 100 mL do produto, duas vezes ao dia, todos os dias. A avaliação dos voluntários foi feita antes do início do ensaio e também aos ao final dos 90 dias.
The gastrointestinal tract of humans is rich in microorganisms which can both be beneficial to host health, preventing and / or treating lactose intolerance, constipation, irritable bowel syndrome, among others, as they can harm, affecting nutrient acquisition and the production of inflammatory mediators. These distinctive roles of intestinal microbiota are so striking that can even influence the development of obesity in some people and may even lead to metabolic syndrome. Probiotics and prebiotics can confer alterations in the microbiota properties, affecting the bacteria growth and their metabolism, and the use of nutrients. Thus, the aim of this study was to evaluate the use of innovative symbiotic fermented milk in obesity and metabolic syndrome indicators such as waist circumference, TGL, HDL-C, blood glucose and blood pressure in patients with a predisposition to this disease, selected according to the diagnostic criteria for metabolic syndrome. For this, four fermented mik were designed, three of them inoculated with a commercial probiotic culture - Bifidobacterium lactis (BL420), added or not by passion fruit peel powder (prebiotic), one of them being prepared with an emulsion of vegetable oil, which has effect on prolonguing satiety; and the fourth product, fermented only with the classical culture Streptococcus thermophilus. Products´ technological properties were evaluated - physico chemical, sensory, microstructure, microbiology, and functional aspects were studied. Finally products were tested in consumers throughout a clinical trial during 12 weeks, period in which the volunteers had to consume 100 mL of the product twice a day, every day. The evaluation of the volunteers was done before the start of the test and also at the end of the 90 days.

Os produtos lácteos probióticos e/ou simbióticos são líderes dentro do mercado de alimentos funcionais e prioridade de pesquisa em diversos países. A presença de bactérias probióticas viáveis e em alto número no produto durante sua vida de prateleira é condição essencial para assegurar o efeito probiótico. As bactérias probióticas apresentam crescimento lento no leite. O uso de co-culturas e a suplementação do leite são técnicas para superar esta dificuldade. O presente trabalho teve como objetivo verificar as condições de crescimento de Lactobacillus rhamnosus e Lactobacillus acidophilus em cultura mista com Streptococcus thermophilus em leite suplementado com hidrolisado de caseína e proteína concentrada de soro em diversas proporções (3,05 a 17,05g/L). Os parâmetros cinéticos velocidade de acidificação, tempo para atingir a velocidade máxima, tempo para atingir pH 5,0 e tempo para atingir pH 4,5 foram analisados, assim como as características físico-quimicas dos leites fermentados e a viabilidade das bactérias probióticas. A metodologia de superfície de resposta foi empregada visando otimizar a suplementação. O desempenho dos probióticos foi comparado com o leite fermentado pelas bactérias normais do iogurte nas mesmas condições. Ambos suplementos afetaram a acidificação do leite, sendo possível reduzir o tempo de fermentação. Em todos os ensaios, o nível de probióticos obtido esteve acima de 106 UFC/mL, valor mínimo de microorganismos viáveis sugerido por diversos autores para obter o efeito benéfico. Foi possível identificar modelos matemáticos preditivos.
Dairy products containing probiotics and/or symbiotics are leader in the functional foods market and, priority in research worldwide. The use of casein hydrolysate and co-cultures are useful to lower fermentation time, and, in addition, it enhances the probiotic stability. The aim of the work was to verify the growing conditions of Lactobacillus rhamnosus and Lactobacillus acidophilus in mixed cultures with Streptococcus thermophiles in milk supplemented with casein hydrolysate and whey protein with different amounts (3,05 to 17,05g/L). Response surface methodology was used to establish adequate relationship of casein hydrolysate and whey protein contents aiming their optimization. The performance of probiotic bacteria was compared with those of yogurt bacteria. The supplements influenced milk acidification. In all experiments, probiotic levels were higher than 106 cfu/mL, the minimal viable number suggested by many authors to have therapeutic benefits. Mathematical models were developed.

Os produtos lácteos probióticos e/ou simbióticos são líderes dentro do mercado de alimentos funcionais e têm prioridade de pesquisa em diversos países. Os resultados deste trabalho mostraram que a qualidade do leite fermentado foi fortemente influenciada tanto pela composição das co-culturas probióticas quanto por diferentes prebióticos, como oligofrutose, polidextrose, maltodextrina e inulina. A cinética de acidificação foi influenciada pela composição das culturas probióticas e pelos ingredientes prebióticos no leite fermentado. A suplementação do leite com a inulina reduziu o tempo de fermentação das co-culturas Streptococcus thermophilus + Lactobacillus acidophilus (St-La); Streptococcus thermophilus + Lactobacillus rhamnosus (St-Lr) e Streptococcus thermophilus + Bifidobacterium lactis (St-Bl), além de melhorar a firmeza do leite fermentado probiótico. Foram também relatadas que as quantidades de ácido linoléico conjugado (CLA) aumentaram no leite fermentado pela co-cultura de S. thermophilus + L. acidophilus, suplementado com maltodextrina. No que diz respeito às contagens, a adição de inulina favorece a viabilidade das bactérias probióticas durante o armazenamento a 4°C além de causar um efeito bifidogênico, in vitro, estimulando o crescimento de B. lactis. Em relação ao estudo metabólico entre as co-culturas homofermentativas (St-La e St-Lb), feitas neste presente trabalho, pode-se dizer que a lactose foi apenas parcialmente fermentada a ácido lático, a galactose foi metabolizada em certa medida, e se formaram diacetil e acetoína em níveis apreciáveis. A acetoína e o diacetil provavelmente foram produzidos pelas atividades da α-acetolactato sintase e da α-acetolactato descarboxilase de S. thermophilus.
Probiotics dairy products and/or symbiotic are leaders in the functional foods market and have the research priority in several countries. The results of this study showed that the quality of fermented milk was strongly influenced by composition of probiotic co-cultures and different prebiotics, such as oligofructose, polydextrose, maltodextrin and inulin. The acidification kinetics was influenced by the composition of the probiotic co-cultures and prebiotic ingredients in the fermented milk. The milk supplementation with inulin reduced the fermentation time of the co-cultures of Streptococcus thermophilus + Lactobacillus acidophilus (St-La); Streptococcus thermophilus + Lactobacillus rhamnosus (St-Lr) and Streptococcus thermophilus + Bifidobacterium lactis (St-Bl), and improved the firmness of the probiotic fermented milk. It has also been observed that the amount of conjugated linoleic acid (CLA) increased in the milk fermented by the co-culture S. thermophilus + L. acidophilus supplemented with maltodextrin. As far as the bacterial counts are concerned, the inulin addition promoted the viability of probiotic bacteria during storage at 4°C and led to a bifidogenic effect, in vitro, stimulating the growth of B. lactis. As regards to the metabolic studies of the homofermentative co-cultures (St-La and St-Lb), studied in this work, it can be said that lactose was only partially fermented to lactic acid, galactose was metabolized to some extent, diacetyl and acetoin formed at appreciable levels. The acetoin and diacetyl were probably produced by the activities of α-acetolactate synthase and α-acetolactate decarboxylase of S. thermophilus.

O principal crescimento na indústria de alimentos funcionais corresponde ao dos produtos probióticos e prebióticos. A literatura mostra efeitos imunomoduladores de certas cepas probióticas, contudo, os resultados são às vezes controversos e os mecanismos implicados ainda são pouco elucidados. Sabe-se, no entanto que algumas cepas de probióticos aumentam significantemente a liberação de IL-10 e γ-INF modulando a resposta imune, além destas respostas serem de forma mais branda relacionada às bactérias Gram-positivas probióticas do que às Gram-positivas patogênicas. O presente trabalho teve como objetivo geral estudar o efeito do leite probiótico fermentado na resposta imune celular em cólon de camundongos BALB/c. Os objetivos específicos foram: (i) determinar o efeito imunomodulador do leite adicionado de probiótico em camundongos normais, (ii) identificar os tipos celulares implicados na resposta imune específica por citometria de fluxo e, (iii) colocalizá-los nos cortes histológicos. Simultaneamente, a análise e a comparação da resistência do probiótico à digestão gastrintestinal in vitro e a produção de metabólitos bioativos de acordo com os deferentes produtos foi realizada. Foram preparados leites nos quais as variáveis estudadas foram a tecnologia empregada para a produção das formulações (a) leite; (b) água, (c) leite não fermentado; (d) leite fermentado; (e) leite fermentado seguido de pasteurização, usando a mesma concentração da cepa comercial Bifidobacterium animalis subsp. lactis HOWARU HN019. O leite desnatado e a água foram usados como controles.
Functional food industry is in expansion mainly due to probiotic and prebiotic products. Studies have shown some probiotic strains develop immune modulation effect, however, these results are controversial and the mechanisms are not been well understood. Although, some probiotic strains increase IL-10 and γ-INF release modulating immune response, this response is weaker in probiotic strains when compared to pathogenic Gram-positive bacteria. The major aim of the present study was to assess the effect of probiotic fermented milk in cellular immune response of Balb/c mice colon. The specific objectives were: (i) to determine the immunomodulation of the milk added of probiotic in normal mice; (ii) to identify the cellular types implied in immune specific response and, (iii) to colocalize them in histological sections. Besides, the analyze and comparation of the probiotic resistance upon in vitro gastrointestinal and bioactive metabolites release in fermented or unfermented bifido milk using the same matrix, probiotic strain and probiotic dose in CFU. mL-1 were conducted. Dairy products were prepared in which variable form of technological appliance were: (i) milk, (ii) water, (iii) unfermented milk, (iv) fermented milk, and (v) fermented and heat treatment milk, all using Bifidobacterium subsp. lactis HOWARU HN019 strain in the same concentration. The skimmed milk and water were used as controls. The immune effects were evaluated by histological sections and the lymphocytic infiltrated was analyzed by flow citometry and histology.

1-4% of pulse fractions including pea protein and fiber, chickpea and lentil flour as well as soy flour and protein concentrate were selected and characterized. As preliminary results the functional properties of pulse ingredients are varied upon their protein content and pH of the food carrier. Orange juice, apple juice, yogurt and two probiotic fermented milk were selected for supplementation. 1% and 2% pulse fractions gave comparable results in terms of turbidity, cloud and visual stability, color and sensory attributes for both orange and apple juices beverages. All supplements improved the acidification rate of yogurt and probiotic cultures, but the highest effects were obtained with probiotic supplementation with lentil and soy flour. As for the main study, skim milk (9.5 % w/v solid content) was supplemented with 1-3% (w/v) lentil flour, pea flour or skim milk powder and they were inoculated with yogurt starter cultures or probiotic (L. rhamnosus). Acid production during the fermentation, the pH, syneresis, color, rheological properties (dynamic oscillation temperature sweep test at 4-50 ˚C), and sensory properties (only for yogurt) were studied after production and 28 days of refrigerated storage.1-3% lentil and pea flour enhanced acid production during yogurt fermentation, but the microbial population (CFU) of both S. thermophilus and L. bulgaricus were in the same range in all lentil and pea flour and skim milk supplemented yogurts, after production. Pea flour supplementation enhanced survival of L. bulgaricus after storage. The pH decreased from 4.5 to 4.1 in lentil flour and from 4.5 to 3.75 in pea flour supplemented yogurts, after 28 days. Syneresis in 1-2% lentil and pea flour supplemented yogurts was higher than other samples. In lentil supplemented yogurts, "a" and "L" values did not significantly differ in all samples and remained constant after 28 days whereas, "b" value increased as a result of supplementation. Pea flour supplementation did not alter redness or greenness of yogurts, but the yellowness was significantly higher than other yogurts. Yogurt with 3% lentil and pea flour had higher storage (G΄) and loss (G˝) moduli in comparison with samples supplemented with 1-3% skim milk and the control yogurt. 1-2% lentil and pea flour supplemented yogurt showed comparable sensory properties in comparison with 1-2% skim milk supplemented and control samples.1-3% lentil and pea flour enhanced acid production during probiotic fermentation, and the CFU's of L. rhamnosus were comparable with non-supplemented control sample after production. After 28 days, the CFU`s of 2% and 3% lentil supplemented probiotic were as high as 1% skim milk supplemented sample and the CFU`s of 3% pea flour supplemented probiotic was the highest followed by 3-2% skim milk and 1-2% pea flour supplemented samples. The pH decreased from 4.50 to 3.90 for lentil flour supplemented probiotics and from 4.50 to 4.04 for pea flour supplemented probiotics, over 28 days. Syneresis in 1-3% lentil and pea flour supplemented probiotic was significantly lower than other samples. All lentil flour supplemented samples had significantly lower "L" values and higher "b" and "a" values in comparison with skim milk supplemented samples. Pea flour supplementation slightly changed the color which was not as light as skim milk supplemented samples and they showed more yellowness in final product after production and storage. Probiotic fermented milk with 1-3% lentil and pea flour showed higher G΄ and G˝ in comparison with other samples.
Des légumineuses tels que des protéines et fibres de pois, farine de pois chiche, de lentille et de soja ont été sélectionnées et caractérisés. Des résultats préliminaires ont montré que des propriétés fonctionnelles ont variés en fonction de la teneur en protéines et du pH des légumineuses employées. Du jus d'orange et de pomme, du yogourt et deux laits fermentés à l'aide de probiotiques ont été supplémentés avec les différentes légumineuses à des taux de 1 à 4%. Les supplémentations à 1 et 2% ont donné des résultats comparables en termes de turbidité, de stabilité, de couleur et d'attributs sensoriels pour les jus d'orange et de pomme. L'addition de légumineuse a permis d'avoir une acidification plus rapide dans les yaourts et les cultures probiotiques, mais le effet le plus important a été obtenu avec farine de lentilles et le soja dans les cultures probiotiques. Comme précédemment, des laits écrémés (9,5% p/v) ont été supplémentés avec 1-3% (p/v) de farine de lentilles, de pois ou de poudre de lait écrémé. Ils ont été inoculés avec des cultures de yogourt, des probiotiques (L.rhamnosus). La production d'acide lors de la fermentation, le pH, la synérèse, la couleur, les propriétés rhéologiques (essai dynamique balayer oscillation de température à 40-50˚C), et les propriétés sensorielles (uniquement pour les yogourts) ont été étudiés après la production et durant 28 jours d'entreposage frigorifique. 1-3% de farine de lentilles ou de pois ont amélioré la production d'acide pendant la fermentation du yogourt, mais les UFC ont les même compte pour les laits suppléments que pour les témoins (lait écrémé). Il est a noter que L. bulgaricus avaient un meilleur taux de survie au jour 28 avec une supplémentation en farine de pois. La diminution du pH dans les yogourts est de 4,5 à 4,1 avec la farine de lentille et de 4,5 à 3,75 avec farine de pois, après 28 jours. La synérèse pour les yogourts supplémentés à 1 et 2% avec de la farine de lentille ou de pois était supérieure d'autres échantillons. Lorsque le taux de supplémentation augmente en farine de lentille ou de pois, il n'y a pas de différence significative pour les valeurs de a alors que la valeur b a augmenté en fonction de la supplémentation.Les yogourts faits de 1 a 3 % farine de lentilles et de pois 1 3% avaient un module élastique (G') et un module visqueux (G˝) plus élevés que les échantillons supplémentés en lait écrémé et que les témoins. Les Yogourts avec 1 à 2% de farine de lentilles et de pois possèdent des propriétés sensorielles comparable a celles des yogourts faits avec 1 a 2% de lait écrémé et celles des témoins. 1-3% de farine de lentilles ou de pois dans des laits avec probiotiques ont amélioré la production d'acide pendant la fermentation, et les UFC de L rhamnosus étaient comparable a ceux des témoins (lait écrémé) après production. Après 28 jours, les UFC pour les échantillons supplémentés avec 2 et 3% de farine de lentille étaient aussi élevées que ceux supplémentés avec 1% de lait écrème et les UFC pour les échantillons supplémentés avec 3% de farine de pois étaient plus élevées que ceux de tous les autres échantillons. Durant les 28 jours de production le pH diminue dans les laits probiotiques contenant de la farine de lentille de 4,50 à 3,90 et pour ceux contenant de la farine de pois de 4,50 à 4,04. La synérèse dans laits probiotiques avec 1 à 3% de farine de lentilles ou de pois a été significativement plus faible que les autres échantillons. Tous les échantillons contenant de farine de lentilles avaient significativement une valeur de L plus bas et des valeurs de b et a plus élevés en comparaison aux échantillons supplémentés en lait écrémé. L'addition de farine de pois a entraîné une modification de couleur b.Les laits probiotiques supplémentés avec 1 a 3 % de farine de lentilles et de pois ont des valeurs de G' et G˝ supérieures aux autres échantillons.

O presente trabalho tem como objetivo estudar o perfil de acidificação e a inter-relação entre Streptococcus thermophilus TAO, Lactobacillus delbrueckii subsp. bulgaricus LB340, Lactobacillus acidophilus LAC, Lactobacillus rhamnosus LBA, Bifidobacterium lactis BL04 como culturas associadas em leite fermentado. Cinco leites fermentados foram preparados, sendo a composição das co-culturas a variável estudada. O perfil de acidificação foi monitorado e os parâmetros cinéticos, calculados. Os produtos foram submetidos às análises físico-químicas e microbiológicas durante o armazenamento a 4°C. As associações em cultura mista provocaram a redução do tempo de fermentação do leite. Durante os 21 dias de armazenamento o pH e a firmeza dos leites fermentados variaram. Streptococcus thermophilus TAO, Bifidobacterium lactis BL04 e Lactobacillus rhamnosus LBA forneceram contagens acima de 106 log UFC/mL, porém Lactobacillus acidophilus LAC e Lactobacillus bulgaricus LB340 foram inibidos em cultura mista, demonstrando dificuldades de crescimento quando associados às demais bactérias ácido-láticas.
The present study aimed to evaluate the acidification kinectic and inter-relation between Streptococcus thermophilus TAO, Lactobacillus delbrueckii subsp. bulgaricus LB340, Lactobacillus acidophilus LAC, Lactobacillus rhamnosus LBA, Bifidobacterium lactis BL04 like association cultures in fermented milk. Five fermented milks were prepared and studied variable analyzed was the co-cultures composition. Acidification was monitored and the kinectic parameters were calculated. The products were submitted to physical chemistry and microbiological analyses during the storage at 4°C. The associations in mixed cultures promoted the reduction of fermentation time of the milks. During 21 days of storage, pH and firmness of fermented milks varied. Streptococcus thermophilus TAO, Bifidobacterium lactis BL04 and Lactobacillus rhamnosus LBA presented counts above 106 log cfu/mL. However, Lactobacillus acidophilus LAC and Lactobacillus bulgaricus LB340 were inhibited in mixed cultures demonstrating that these strains had difficulty to grow when in associated cultures with lactic acid bacteria.

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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.

O objetivo deste trabalho foi desenvolver uma bebida láctea fermentada preparada com 60% de leite, 32% de soro lácteo e 8% de açúcar, utilizando micro-organismos probióticos. Foram desenvolvidas 3 formulações, com 1% (A), 3% (B) e 5% de semente de chia (Salvia hispanica L.). Foram realizadas análises físico-químicas na formulação padrão e nas formulações A, B e C durante 0, 7 e 14 dias; análise sensorial nas formulações A, B, e C nos tempos 0, 7 e 14 dias e avaliação microbiológica. O teor de umidade nas 3 formulações apresentou diferença significativa aos 0 e 7 dias de estocagem (p < 0,05). Os teores de cinzas, proteínas, lipídeos e a viscosidade da bebida aumentaram significativamente à medida que se aumentou a proporção de chia na bebida. Foi observada uma redução gradativa do pH e aumento da acidez titulável em função do tempo de estocagem da bebida. A bebida láctea obteve boa aceitação sensorial, mas foi observada uma diminuição nas notas da escala hedônica e intenção de consumo durante o período de estocagem. A contagem de bactérias probióticas teve redução de aproximadamente 1 ciclo logarítmico por semana. A formulação preferida no teste de ordenação de preferência foi a formulação A.
The aim of this study was to develop a fermented dairy beverage made with 60% milk, 32% whey and 8% sugar, using probiotic microorganisms. Three formulations, with 1% (A), 3% (B) and 5% chia seed (Salvia hispanica L.) have been developed. Physico-chemical analyzes on standard formulation and the formulations A, B and C for 0, 7 and 14 days were made; sensory analysis in formulations A, B, and C for times 0, 7 and 14 days and microbiological evaluation The moisture content of the three formulations showed a significant difference at 0 and 7 days of storage (p <0.05). Contents of ash, protein, lipids and viscosity increased significantly as it increased the proportion of chia. A gradual reduction of pH and increase in acidity due to the storage time of the beverage was observed. The product has obtained good acceptability, but a decrease was observed in the global acceptability and intention of consumption during the storage period. The probiotic bacteria count was reduced by about 1 log cycle per week. The preferred formulation of the ranking preference test was the formulation A.

CNPq
A crescente procura dos consumidores por alimentos que, além das funções básicas de nutrição, exercem efeitos benéficos à saúde, tem estimulado os pesquisadores e as indústrias alimentícias a desenvolverem produtos lácteos funcionais. Nesta classe, atenção especial tem sido dada ao desenvolvimento de iogurtes com propriedades probióticas e prebióticas, denominados de simbióticos. Inúmeros mecanismos têm sido propostos para tentar explicar os benefícios do consumo dos produtos lácteos simbióticos, dentre eles a capacidade que os micro-organismos probióticos têm de produzir ácido linoleico conjugado. O consumo deste composto tem sido associado a efeitos benéficos à saúde, como propriedades anticarcinogênicas, modulação do sistema imune, diminuição da aterosclerose, entre outros. O presente trabalho teve por objetivo geral a produção e caracterização de um iogurte simbiótico sabor banana obtido a partir dos leites de cabra e de vaca, das culturas probióticas Lactobacillus acidophilus e Bifidobacterium animalis subps lactis Bb-12 e polpa de banana verde como fonte de amido resistente. Para atingir este objetivo, as seguintes etapas foram realizadas: revisão bibliográfica sobre ácido linoleico conjugado, por serem o leite e seus derivados as principais fontes destes ácidos graxos; análise da composição centesimal e das propriedades físico-químicas do leite de vaca, do leite de cabra, da banana verde, da polpa de banana verde e dos iogurtes produzidos; verificação da viabilidade dos diferentes tratamentos de iogurte como carreadores de micro-organismo probiótico através da enumeração de Lactobacillus acidophilus e de Bifidobacterium animalis subps lactis Bb-12 durante a vida útil dos produtos (tempo 0, 15, 30 e 45 dias); quantificação dos micro-organismos Lactobacillus delbruecki subsp. bulgaricus e Streptococcus salivarius subps. thermophilus empregados como cultura iniciadora na elaboração dos iogurtes, durante a vida útil dos produtos (tempo 0, 15, 30 e 45 dias); estudo do perfil de textura (dureza, coesividade, elasticidade e gomosidade) e viscosidade aparente dos diferentes tratamentos de iogurte durante a vida útil (0, 15, 30 e 45 dias) e determinação da cor (parâmetro L* e coordenadas a* e b*) das diferentes formulações de iogurtes durante a vida útil (0, 15, 30 e 45 dias), um estudo de mercado e por fim, análise sensorial dos iogurtes elaborados. As seis formulações de iogurte atenderam às exigências da legislação vigente para as características físico-químicas, para a composição centesimal e para as contagens microbiológicas. Ainda, foram observadas diferenças significativas na composição centesimal, nas propriedades físico-químicas, nos perfis de cor e textura e na análise sensorial dos iogurtes elaborados com leite de vaca daqueles elaborados com leite de cabra.
The growing demand from consumers for foods that beyond basic nutritional functions, exert beneficial health effects has stimulated researchers and the food industry to develop functional dairy products. In this class, special attention has been given to the development of yoghurt with probiotic and prebiotic properties, called symbiotic. Numerous mechanisms have been proposed to explain the benefits of consuming dairy symbiotic products, including the ability of probiotic microorganisms on conjugated linoleic acid production. The use of this compound has been associated with beneficial health effects, such as anticarcinogenic properties, modulation of the immune system, reduction of atherosclerosis, among others. The overall aim of this study was the production and characterization of a symbiotic banana flavored yogurt obtained from cow and goat milk, probiotic cultures Lactobacillus acidophilus and Bifidobacterium animalis subsp lactis Bb-12 and green banana pulp as resistant starch source. To achieve this goal, the following steps were carried out: literature review about conjugated linoleic acid, analysis of the chemical composition and physicochemical properties of cow's milk, goat milk, green banana, green banana pulp and yoghurt produced; checking the viability of the probiotic microorganism Lactobacillus acidophilus and Bifidobacterium animalis subsp lactis Bb-12 by enumeration during the useful life of the products (time 0, 15, 30 and 45 days); quantification of the microorganisms Lactobacillus delbruecki subsp. bulgaricus and Streptococcus salivarius subps. thermophilus as starter culture employed in the preparation of yoghurt during the product lifetime (time 0, 15, 30 and 45 days); profile study of texture (hardness, cohesiveness, elasticity or gumminess) and apparent viscosity of different treatments during yogurt lifetime (0, 15, 30 and 45 days) and determining color (parameter L * and a * b * coordinates) of different formulations of yogurts during the lifetime (0, 15, 30 and 45 days), market study and finally, the sensory analysis of yoghurt prepared. The six yogurt formulations achieved the legal requirements for the physico-chemical properties, for the centesimal composition and for microbiological enumeration. Still, significant differences were observed in the centesimal composition, the physico-chemical properties, color and texture profiles and sensory analysis of yogurt made with cow’s milk from those made with goat’s milk.

The aim of this project was to study the identity of probiotic lactobacilli in fermented milk products from the United Kingdom/European markets during their survival during shelf-life. This in vitro study was also aimed at undertaking studies on some of the physiological probiotic criteria, such as resistance to stomach/intestine conditions and also possible functional properties of the isolates, such as antimicrobial activities, antibiotic resistance/susceptibility and antibiotic resistance genes, biofilm formation and production of conjugated linoleic acid (CLA). Primarily, a comparative study was carried out on selectivity of MRS-Clindamycin, MRS-Sorbitol and MRS-IM Maltose, to select the right medium for enumeration of probiotic Lactobacillus. Based on selectivity of medium for recovery of the targeted lactobacilli and also simplicity of preparation, MRS-Clindamycin was chosen as the best medium for enumeration of probiotic Lactobacillus in fermented milks. The results of enumeration of lactobacilli showed that 22 out of a total 36 tested products contained more than 106 colony forming units/g at the end of their shelf-life, which comply with the recommended minimum therapeutic level for probiotics. Rep-PCR using primer GTG-5 was applied for initial discrimination of isolated strains, and isolates, which presented different band profile, were placed in different groups. The isolated Lactobacillus spp. were identified mainly as Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus paracasei by analysis of partial sequences of the 16S ribosomal RNA and rpoA genes. In order to characterize the isolates for probiotic properties, this study was focused on six Lactobacillus isolates along with two commercial Lactobacillus cultures from Chr. Hansen (Lactobacillus acidophilus La5 and Lactobacillus casei C431) and three Lactobacillus type strains (Lactobacillus casei subsp. casei, Lactobacillus paracasei subsp. paracasei and Lactobacillus acidophilus) which were purchased from NCIMB. The stomach and intestine conditions were mimiced using a batch culture fermentation system, and the combined effects of pH, enzymes and bile salts on survival of tested isolates was tested. The tested isolates were able to survive at low pH environment and also high concentrations of bile salts of the upper digestive tract. The potential of tested isolates for biofilm formation was determined in different conditions of nutritional and physiological stresses. The capability of tested isolates to produce biofilm in nutrient rich medium was recorded. However, the growth limitation, such as nutrient shortage in diluted media and also using inulin rather than glucose in synthetic medium, did not induce biofilm formation. Antimicrobial activities of tested bacteria against indicator bacteria namely Escherichia coli NCTC 12900, Salmonella enterica serovar Typhimurium DT124 and Salmonella enterica serovar Enteritidis PT4 and Lactobacillus delbruckii subsp. bulgaricus were studied. The production of organic acids and bacteriocin was considered as key mechanisms for antimicrobial activity of tested strains. Screening the isolates competence for production of CLA demonstrated that this feature is species dependent and also entirely related to the level of initial linoleic acid in the medium. Eleven tested isolates were also assessed for their antibiotic resistance profile by determination of minimum inhibitory concentration (MIC). The acquired resistance to cefoxitin, ceftriaxone, chloramphenicol, erythromycin, gentamycin, kanamycin, lincomycin, streptomycin, tylosin tartarate, tetracycline and vancomycin was observed in all tested isolates. Also their genetic background of antibiotic resistance genes was studied by PCR reactions and none of the tested isolates showed positive bands for investigated resistance genes.

O objetivo deste estudo foi avaliar o efeito da inclusão de leite em pó desnatado e de açúcar sobre a viabilidade de microrganismos probióticos e a qualidade do leite fermentado desnatado. Para isso, foram constituídos 8 tratamentos, a saber: 1. leite desnatado (LD) e padronizado com 0,2% de gordura (LD 0,2%G); 2. LD 0,2%G + 5% de leite em pó (Lp); 3. LD 0,2%G + 10% de Lp; 4. LD 0,2%G + 15% de Lp; 5. LD 0,2%G + 5% de Lp + 10% de açúcar (A); 6. LD 0,2%G + 10% de Lp + 10% de A; 7. LD 0,2%G + 15% de Lp + 10% de A; 8. LD 0,2%G + 10% de A. Foi preparada a cultura láctea mista DVS ABT-4 (Chr-Hansen) contendo Streptococcus thermophilus, Bifidobacterium animalis e Lactobacillus acidophilus, que foi aplicada em todos os tratamentos para a fermentação e produção do leite fermentado. O produto foi então envasado e armazenado a 5ºC para as análises subseqüentes. Os leites fermentados foram submetidos à avaliação físico-química, microbiológica e sensorial nos dias 1, 10 e 21 após a fabricação. Os resultados mostraram que todos os tratamentos permaneceram de acordo com a legislação brasileira no que diz respeito aos aspectos físico-químicos, além de os mesmos terem permanecido constantes durante os 21 dias de armazenamento, com exceção do pH, que diminuiu nos dias seguintes a fabricação em decorrência da pós-acidificaçao dos leites fermentados. Quanto às características microbiológicas, observou-se maiores contagens nos tratamentos com maior teor de leite em pó (tratamentos 3, 4, 6 e 7), principalmente para as bactérias probióticas, com diferença estatisticamente significativa entre todos os tratamentos. Entretanto, a adição de açúcar não interferiu nas contagens de probióticos dos leites fermentados. As contagens microbiológicas permaneceram constantes durante todos os 21 dias de armazenamento. O tratamento 6 (LD 0,2%G + 10% de Lp + 10% de A) foi o que obteve melhores notas na avaliação sensorial nos dias 1, 10 e 21 de armazenamento, indicando ser esta formulação a melhor dentre as estudadas para a aceitação do produto com maior viabilidade de bactérias probióticas.
The aim of this study was to evaluate the effect of skimmed milk powder and sugar inclusion on the viability of probiotic microorganisms and on the quality of skimmed fermented milk. Eight different treatments were prepared: 1. Skimmed and standardized milk (SM) at 0,2% of fat (SM 0,2%F); 2. SM 0,2%F + 5% of milk powder (MP); 3. SM 0,2%F + 10% MP; 4.SM 0,2%F + 15% MP; 5. SM 0,2%F + 5% MP + 5% of sugar (S); 6. SM 0,2%F + 10% MP + 5% S; 7. SM 0,2%F + 15%MP + 10%S; 8. SM 0,2%F + 10%S. The mixed lactic culture DVS ABT-4 (Chr-Hansen) containing Streptococcus thermophilus, Bifidobacterium animalis and Lactobacillus acidophilus was prepared and placed in all the treatments for the fermented milk fermentation and production. The product was bottled and stored at 5°C for the subsequent analyses. The fermented milks were submitted to physical-chemical, microbiological and sensory analyses on the 1st, 10th and 21st days after production. Results for the physic-chemical analyses showed that all the treatments comprised the requirements as adopted in Brazilian regulations. All the physical-chemical variables were constant during the 21 days of storage, except for the pH, which decreased during storage because of the fermented milks post-acidification. For the microbiological analyses, greater numbers, especially the probiotic bacteria, were obtained in treatments with higher milk powder content (treatments 3, 4, 6 and 7), with significant statistical differences among all the treatments. However, the sugar addition did not interfere with the recovery of probiotic bacteria in the fermented milks. The microbiological data were constant during the 21 days of storage. Results obtained in the sensory analyses showed that treatment 6 was the one with higher grades on days 1, 10 and 21 of storage, hence indicating that this composition is the best one among those studied for acceptability of the product with greater viability of probiotic bacteria.

Orientador: Ana Lúcia Barretto Penna
Banca: Susana Marta Isay Saad
Banca: Crispin Humberto Garcia Cruz
Resumo: Os leites fermentados probióticos são líderes no mercado de alimentos funcionais e prioridade de pesquisa em diversos países. A presença de bactérias probióticas viáveis e em alto número no produto durante sua vida de prateleira é condição essencial para assegurar o efeito probiótico. As qualidades tecnológica e funcional de leites fermentados são importantes para sua aceitação pelo consumidor e podem ser afetadas pelos aditivos adicionados antes da fermentação dos produtos. Neste trabalho, o objetivo foi avaliar o efeito da adição de edulcorantes, em diferentes concentrações, na produção de leites fermentados por uma cultura mista composta por Streptococcus thermophilus e Lactobacillus bulgaricus (Rich) e uma cultura pura composta por Lactobacillus acidophilus (La-5). As bases lácteas para preparação dos leites fermentados obtidas antes da fermentação foram submetidas às determinações dos teores de sólidos totais, proteínas, cinzas, gordura, acidez titulável e valor calórico. Nos leites fermentados foram avaliados: a viabilidade das bactérias láticas, a pós-acidificação, a sinérese e a capacidade de retenção de água (CRA), durante 28 dias de estocagem refrigerada. As características físico-químicas dos leites fermentados atenderam aos requisitos legais e apresentaram diferenças somente nos teores de sólidos totais, cinzas e valor calórico nos produtos elaborados com sacarose. O tipo e a concentração de edulcorante afetaram a fermentação da cultura La-5 e a acidez titulável. No entanto, não influenciaram o tempo de fermentação da cultura Rich, a viabilidade de S. thermophilus, L. bulgaricus e L. acidophilus, a sinérese e a CRA dos leites fermentados elaborados. O tipo de cultura influenciou o tempo de fermentação do leite, sendo superiores os tempos obtidos para La-5 e a acidez titulável, com valores superiores para a cultura... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Fermented probiotic dairy products are leaders in the functional food markets and priority for researching worldwide. The presence of probiotic bacteria viable at high level during shelf-life of the product is essential to ensure the probiotic effect. The technological and functional qualities of fermented milks are important for their acceptance by the consumers and they can be affected by the addition of ingredients before fermentation. The aim of this research was to evaluate the use of sweeteners, in different concentrations, in the production of fermented milks using a mixed culture composed of Streptococcus thermophilus and Lactobacillus bulgaricus (Rich), or a pure culture composed of Lactobacillus acidophilus (La-5). The milk bases obtained before fermentation were submitted to determinations of total solids, protein, ash, fat, acidity contents and caloric value. The viability of lactic bacteria, post-acidification, syneresis and waterholding capacity (WHC) of fermented milks were examined during 28 days of cold storage. The physicochemical characteristics of fermented milks were according to legal requirements and showed differences only on the contents of total solids, ash and caloric value in the products formulated with sucrose. The type and concentration of sweeteners affected the fermentation of La-5 culture and titratable acidity. However they did not influence fermentation time of Rich culture, viability of S. thermophilus, L. bulgaricus and L. acidophilus, syneresis and WHC of fermented milks. The culture type affected the fermentation time, which was higher for La-5 culture, and acidity, which was higher for Rich culture, however, they did not influence the syneresis and WHC of fermented milks. All kinds and concentrations of sweeteners did not influence the technological quality of products, so all of them can be used to produce lowcalorie fermented milks.
Mestre

Subprodutos de frutas são rotineiramente descartados pelas indústrias. Porém, são ricos em compostos bioativos, podendo ser utilizados como ingredientes em produtos funcionais, promovendo a saúde e minimizando o impacto ambiental. O objetivo deste trabalho foi estudar o potencial funcional de subprodutos desidratados de acerola e de laranja, como fontes de compostos fenólicos, e desenvolver leites fermentados probióticos adicionados deste resíduo, avaliando suas características físico-químicas durante o armazenamento sob refrigeração (28 dias; 4 ± 1 °C), bem como o impacto das condições gastrointestinais sobre os flavonoides e as cepas probióticas. Os subprodutos foram obtidos em indústrias de processamento de frutas do estado de São Paulo, e foram realizadas as seguintes análises para caracterizá-los: composição centesimal, teores de vitamina C, minerais, fibras alimentares, compostos fenólicos totais e proantocianidinas, capacidade antioxidante in vitro e perfil cromatográfico de flavonoides (CLAE). Foram elaboradas quatro formulações de leites fermentados: F0 (controle), sem adição de resíduo de acerola (RA); F2, com 2% de RA; F5, com 5% de RA; F10, com 10% de RA. Adicionou-se a cultura probiótica ABT-4 nos produtos, constituída de duas cepas probióticas: Bifidobacterium animalis subsp lactis Bb-12 e Lactobacillus acidophilus La-5, além da cultura starter Streptococcus thermophilus. As seguintes análises foram realizadas com as formulações de leites fermentados, durante o armazenamento sob refrigeração (28 dias, 4 ± 1 °C): composição centesimal, pH, acidez, viabilidade dos microrganismos, teor de compostos fenólicos totais (CF), cor e textura instrumentais. Além disso, os leites fermentados foram submetidos a condições gastrointestinais simuladas in vitro, para avaliação do impacto na viabilidade das cepas probióticas e nos compostos fenólicos. O RA mostrou-se excelente fonte de vitamina C (605 mg/100 g b.u.), além de apresentar melhor capacidade antioxidante in vitro do que o RL. Proantocianidinas foram encontradas apenas no RA, na concentração de 617 µg EC/g b.s. O teor de compostos fenólicos totais do RA (3240 µg EAG/100 g b.s.) foi 3,6 vezes maior que o do RL. Os principais compostos fenólicos encontrados no RA foram: derivados de quercetina, procianidina B1, rutina, e derivados de caempferol. No RL, foram identificados: naringenina, sinensetina, homorientina, isovitexina e derivados de ácido clorogênico. Os subprodutos estudados apresentaram elevados teores de fibras totais (acima de 60%) e proteínas totais (RA: 10,4%; RL: 9,9%), além de reduzido teor de lipídeos totais (RA: 1,6%; RL: 2,6%). Os principais minerais identificados em ambos os resíduos foram: potássio, magnésio, cálcio e fósforo. Quanto às quatro formulações de leites fermentados, estas apresentaram baixo teor de lipídeos totais (menor que 1%), e o teor de proteínas totais variou entre 3,9 e 5,1 g/100 g, estando de acordo com a legislação vigente para este tipo de produto. O pH das formulações F0 (controle) e F2 manteve-se estável significativamente (p > 0,05) ao longo do período de armazenamento sob refrigeração (28 dias; 4 ± 1 °C), e das outras formulações sofreu pequena queda, mesmo assim mantendo-se acima de 4,5. A acidez das formulações, que variou entre 0,92 a 1,28 mg de ácido lático/g, aumentou entre os dias 1 e 14 de armazenamento, depois se manteve até o final da vida de prateleira. O RA não interferiu de maneira negativa nas populações de microrganismos analisadas durante o armazenamento, já que as formulações F2, F5 e F10 mantiveram suas populações em torno de 8 log UFC/g. Quanto ao teor de CF, as amostras diferiram significativamente entre si (p < 0,05), sendo que F0 apresentou teor em torno de 5 vezes inferior a F10 (21,13 e 101,13 µg EAG/100 g, respectivamente, no dia 1). A cor dos produtos se manteve até o final da vida de prateleira, e diferiram significativamente (p < 0,05) entre si. O RA influenciou pouco nos parâmetros de textura dos leites fermentados, mas a formulação controle foi a única que perdeu adesividade. Após a fase gástrica da digestão simulada in vitro, no 7° dia de armazenamento, as populações de bactérias probióticas diminuíram drasticamente (quedas em torno de 3 a 5 log UFC/g), e após a fase entérica não foram detectadas contagens. Por outro lado, os flavonoides encontrados nos leites fermentados adicionados de RA aumentaram em torno de 2 a 5 vezes, após a fase gástrica, mantendo-se ou sofrendo pequena queda após fase entérica. Estes resultados mostram que o pó de subprodutos de acerola é um valioso ingrediente a ser utilizado em alimentos funcionais, pois é rico em vitamina C, fibras e compostos fenólicos, agregando valor nutricional, além de servir como antioxidante natural. Seus flavonoides parecem ser altamente resistentes aos ácidos e sais da digestão, podendo, assim, exercer efeitos positivos sobre a saúde.
Fruits by-products are routinely discarded by industries. However, they are rich in bioactive compounds, and can be used as ingredients in functional foods, promoting health and minimizing environmental impact. The objective of this study was to investigate the functional potential of acerola and orange dehydrated by-products, as sources of phenolic compounds, and to develop probiotic fermented milks suplemented with this residues, evaluating its physico-chemical characteristics during refrigerated storage (28 days, 4 ± 1 °C), as well as the impact of gastrointestinal conditions on flavonoids and probiotic strains. The by-products were obtained from fruit processing industries of São Paulo, and the following analyzes were performed to characterize them: contents of moisture, ash, lipids, proteins, vitamin C, minerals, dietary fibers, total phenolic compounds and proanthocyanidins, antioxidant capacity in vitro and flavonoids chromatographic profile (HPLC). Were elaborated four formulations of fermented milks: F0 (control), without addition of acerola residue (AR); F2, with 2% AR; F5, with 5% AR; F10, with 10% AR. Was used the probiotic culture ABT-4, composed of two probiotic strains, Bifidobacterium animalis subsp lactis Bb-12 and Lactobacillus acidophilus La-5, in addition to the starter culture Streptococcus thermophilus. During the refrigerated storage (28 days, 4 ± 1 °C), the following analyzes were performed with the fermented milks: contents of moisture, ash, lipids and proteins, pH, acidity, viability of microorganisms, total phenolic compounds (PC), instrumental color and texture. In addition, the fermented milks were submitted to in vitro simulated gastrointestinal conditions to evaluate the impact on the viability of probiotic strains and phenolic compounds. AR presented excellent content of vitamin C (605 mg/100 g), in addition to presenting better antioxidant capacity in vitro than orange residue (OR). Proanthocyanidins were found only in AR (617 µg CE/g). The PC content of AR (3240 µg GAE/100 g) was 3.6 higher than in OR. The phenolic compounds identified in AR were quercetin-3-rhamnoside, rutin and others quercetin derivatives, procyanidin B1 and kaempferol derivatives. In OR, were identified naringenin, sinensetin, homorientin, isovitexin and chlorogenic acid derivatives. The by-products studied showed high total fibers content (above 60%) and total proteins (AR: 10.4%, OR: 9.9%), as well as reduced total lipids content (AR: 1.6%; OR: 2.6%). Both residues showed high levels of potassium, calcium, magnesium and phosphorus. The four formulations of fermented milks presented low total lipids content (below 1%), and the total proteins content ranged from 3.9 to 5.1 g/100 g, being in agreement with the legislation. The pH of F0 (control) and F2 formulations remained stable (p > 0.05) throughout the refrigerated storage period (28 days, 4 ± 1 °C), and the other formulations showed a small decreased, even thus remaining above 4.5. The acidity of the formulations, ranging from 0.92 to 1.28 mg of lactic acid/g, increased between days 1 and 14 of storage, then remained until the end of shelf life. The AR did not negatively interfere in the populations of microorganisms analyzed during storage, since the formulations F2, F5 and F10 maintained their populations around 8 log CFU/g. Regarding PC content, the samples differed significantly (p < 0.05), with F0 being about 5 lower than F10 (21.13 and 101.13 µg GAE/100 g, respectively, in the day 1). The instrumental color of the products remained until the end of shelf life, and differed significantly (p < 0.05) from each other. The AR influenced a little in the texture parameters of the fermented milks, but the control formulation was the only one that lost adhesiveness. After the gastric phase of the simulated digestion in vitro, on the 7th day of storage, the populations of probiotic bacteria decreased dramatically (of 3 to 5 log CFU/g), and after the enteric phase no colonies were detected. On the other hand, the flavonoids found in the fermented milks that were suplemented with AR increased from 2 to 5 times, after the gastric phase, maintaining or suffering small decreased after enteric phase. These results show that acerola by-products powder is a valuable ingredient to be used in functional foods because it is rich in vitamin C, dietary fibers and phenolic compounds, adding nutritional value, and serving as a natural antioxidant. Its flavonoids appear to be highly resistant to the acids and salts of digestion and can thus have positive effects on health.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Os leites fermentados probióticos são líderes no mercado de alimentos funcionais e prioridade de pesquisa em diversos países. A presença de bactérias probióticas viáveis e em alto número no produto durante sua vida de prateleira é condição essencial para assegurar o efeito probiótico. As qualidades tecnológica e funcional de leites fermentados são importantes para sua aceitação pelo consumidor e podem ser afetadas pelos aditivos adicionados antes da fermentação dos produtos. Neste trabalho, o objetivo foi avaliar o efeito da adição de edulcorantes, em diferentes concentrações, na produção de leites fermentados por uma cultura mista composta por Streptococcus thermophilus e Lactobacillus bulgaricus (Rich) e uma cultura pura composta por Lactobacillus acidophilus (La-5). As bases lácteas para preparação dos leites fermentados obtidas antes da fermentação foram submetidas às determinações dos teores de sólidos totais, proteínas, cinzas, gordura, acidez titulável e valor calórico. Nos leites fermentados foram avaliados: a viabilidade das bactérias láticas, a pós-acidificação, a sinérese e a capacidade de retenção de água (CRA), durante 28 dias de estocagem refrigerada. As características físico-químicas dos leites fermentados atenderam aos requisitos legais e apresentaram diferenças somente nos teores de sólidos totais, cinzas e valor calórico nos produtos elaborados com sacarose. O tipo e a concentração de edulcorante afetaram a fermentação da cultura La-5 e a acidez titulável. No entanto, não influenciaram o tempo de fermentação da cultura Rich, a viabilidade de S. thermophilus, L. bulgaricus e L. acidophilus, a sinérese e a CRA dos leites fermentados elaborados. O tipo de cultura influenciou o tempo de fermentação do leite, sendo superiores os tempos obtidos para La-5 e a acidez titulável, com valores superiores para a cultura...
Fermented probiotic dairy products are leaders in the functional food markets and priority for researching worldwide. The presence of probiotic bacteria viable at high level during shelf-life of the product is essential to ensure the probiotic effect. The technological and functional qualities of fermented milks are important for their acceptance by the consumers and they can be affected by the addition of ingredients before fermentation. The aim of this research was to evaluate the use of sweeteners, in different concentrations, in the production of fermented milks using a mixed culture composed of Streptococcus thermophilus and Lactobacillus bulgaricus (Rich), or a pure culture composed of Lactobacillus acidophilus (La-5). The milk bases obtained before fermentation were submitted to determinations of total solids, protein, ash, fat, acidity contents and caloric value. The viability of lactic bacteria, post-acidification, syneresis and waterholding capacity (WHC) of fermented milks were examined during 28 days of cold storage. The physicochemical characteristics of fermented milks were according to legal requirements and showed differences only on the contents of total solids, ash and caloric value in the products formulated with sucrose. The type and concentration of sweeteners affected the fermentation of La-5 culture and titratable acidity. However they did not influence fermentation time of Rich culture, viability of S. thermophilus, L. bulgaricus and L. acidophilus, syneresis and WHC of fermented milks. The culture type affected the fermentation time, which was higher for La-5 culture, and acidity, which was higher for Rich culture, however, they did not influence the syneresis and WHC of fermented milks. All kinds and concentrations of sweeteners did not influence the technological quality of products, so all of them can be used to produce lowcalorie fermented milks.

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Bisco Inc., Il, Usa
Nas últimas décadas, tem se observado uma mudança substancial no hábito dietético das crianças, com um aumento crescente no consumo de produtos comercialmente disponíveis no mercado brasileiro, como os leites fermentados. Diante dessa realidade, o objetivo do presente trabalho foi avaliar o pH, a capacidade tampão, conteúdo de flúor e de cálcio de alguns leites fermentados, bem como avaliar a acidogenicidade e a concentração dos íons flúor, cálcio, fósforo, carboidratos álcali-solúveis no biofilme dental formado in situ, além da desmineralização do esmalte dental bovino através do teste de microdureza superficial. Para análise bioquímica, foram utilizadas 6 marcas de leite fermentado desnatado adoçado (Parmalat®-uva, Chamyto®, Paulista®, Batavito®, Yakult®, Vigor Club®). O pH das amostras variou entre 3.51 e 3.87; a capacidade tampão variou de 470.8 a 804.2 μL da NaOH 1 mol L-1; a concentração do íon flúor variou de 0.027 a 0.958 μg F/g e a concentração de cálcio apresentou uma variação de 0.4787 a 0.8177 mg Ca/g. Para o estudo in situ, selecionou-se 2 marcas de leite fermentado e dez voluntários que receberam dispositivos palatinos contendo 4 blocos de esmalte dental bovino durante três fases de 14 dias cada. Os resultados mostraram que as concentrações dos íons (F, Ca e P) foram significantemente maiores quando se realizou o tratamento C (Batavito®) em comparação aos tratamentos A (Yakult®) e B (sacarose 20%) (p<0,05), os quais não diferiram estatisticamente entre si. Os dois grupos experimentais (Batavito® e Yakult®) tiveram perdas maiores que o controle (Sacarose 20%), embora não tenham apresentado diferença significante entre si.
In the last decades, it has been observed a substantial change in the children's dietary habit, with an increasing in the consumption of products commercially available in the Brazilian market, such as the fermented milks. Due to that reality, the aim of the present study was to evaluate the pH, buffering capacity, fluoride and calcium content of some fermented milks, as well as evaluating the concentration of the fluoride, calcium, phosphate, and alkali-soluble carbohydrates and the acidogenicity of the in situ dental plaque, besides the demineralization of the bovine dental enamel through the superficial microhardness test. For biochemical analysis, 6 brands of fermented milk were used (Parmalat®-grape, Chamyto®, Paulista®, Batavito®, Yakult®, Vigor Club®). The pH of the samples ranged from 3.51 to 3.87; the buffering capacity from 470.8 to 804.2 μL of NaOH 1 mol L-1; the concentration of the ion fluoride from 0.027 to 0.958 μg F/g and the concentration of calcium from 0.4787 to 0.8177 mg Ca/g. For the in situ study, it were selected 2 fermented milks and ten volunteers who received palatine devices containing 4 blocks of bovine dental enamel during three phases of 14 days each. The results showed that the ionic concentrations (F, Ca and P) were significantly larger when the treatment C (Batavito®) took place in comparison to the treatments A (Yakult®) and B (sucrose 20%) (p <0,05), which did not differ significantly amongst themselves. Batavito® and Yakult® had larger losses than the control (Sucrose 20%), although they have not presented significant difference amongst themselves. Considering the results of this study, it could be concluded that although the fluoride and calcium ions had been verified in the fermented milks the commercial brands Batavito® and Yakult® provoked demineralization of the dental enamel.

Orientador: Cleide Cristina Rodrigues Martinhon
Banca: Alberto Carlos Botazzo Delbem
Banca: Fabian Calixto Fraiz
Resumo: Nas últimas décadas, tem se observado uma mudança substancial no hábito dietético das crianças, com um aumento crescente no consumo de produtos comercialmente disponíveis no mercado brasileiro, como os leites fermentados. Diante dessa realidade, o objetivo do presente trabalho foi avaliar o pH, a capacidade tampão, conteúdo de flúor e de cálcio de alguns leites fermentados, bem como avaliar a acidogenicidade e a concentração dos íons flúor, cálcio, fósforo, carboidratos álcali-solúveis no biofilme dental formado in situ, além da desmineralização do esmalte dental bovino através do teste de microdureza superficial. Para análise bioquímica, foram utilizadas 6 marcas de leite fermentado desnatado adoçado (Parmalat®-uva, Chamyto®, Paulista®, Batavito®, Yakult®, Vigor Club®). O pH das amostras variou entre 3.51 e 3.87; a capacidade tampão variou de 470.8 a 804.2 μL da NaOH 1 mol L-1; a concentração do íon flúor variou de 0.027 a 0.958 μg F/g e a concentração de cálcio apresentou uma variação de 0.4787 a 0.8177 mg Ca/g. Para o estudo in situ, selecionou-se 2 marcas de leite fermentado e dez voluntários que receberam dispositivos palatinos contendo 4 blocos de esmalte dental bovino durante três fases de 14 dias cada. Os resultados mostraram que as concentrações dos íons (F, Ca e P) foram significantemente maiores quando se realizou o tratamento C (Batavito®) em comparação aos tratamentos A (Yakult®) e B (sacarose 20%) (p<0,05), os quais não diferiram estatisticamente entre si. Os dois grupos experimentais (Batavito® e Yakult®) tiveram perdas maiores que o controle (Sacarose 20%), embora não tenham apresentado diferença significante entre si.
Abstract: In the last decades, it has been observed a substantial change in the children's dietary habit, with an increasing in the consumption of products commercially available in the Brazilian market, such as the fermented milks. Due to that reality, the aim of the present study was to evaluate the pH, buffering capacity, fluoride and calcium content of some fermented milks, as well as evaluating the concentration of the fluoride, calcium, phosphate, and alkali-soluble carbohydrates and the acidogenicity of the in situ dental plaque, besides the demineralization of the bovine dental enamel through the superficial microhardness test. For biochemical analysis, 6 brands of fermented milk were used (Parmalat®-grape, Chamyto®, Paulista®, Batavito®, Yakult®, Vigor Club®). The pH of the samples ranged from 3.51 to 3.87; the buffering capacity from 470.8 to 804.2 μL of NaOH 1 mol L-1; the concentration of the ion fluoride from 0.027 to 0.958 μg F/g and the concentration of calcium from 0.4787 to 0.8177 mg Ca/g. For the in situ study, it were selected 2 fermented milks and ten volunteers who received palatine devices containing 4 blocks of bovine dental enamel during three phases of 14 days each. The results showed that the ionic concentrations (F, Ca and P) were significantly larger when the treatment C (Batavito®) took place in comparison to the treatments A (Yakult®) and B (sucrose 20%) (p <0,05), which did not differ significantly amongst themselves. Batavito® and Yakult® had larger losses than the control (Sucrose 20%), although they have not presented significant difference amongst themselves. Considering the results of this study, it could be concluded that although the fluoride and calcium ions had been verified in the fermented milks the commercial brands Batavito® and Yakult® provoked demineralization of the dental enamel.
Mestre

O tratamento antimicrobiano pode destruir o equilíbrio da microbiota gastrintestinal, podendo induzir sintomas clínicos, principalmente a diarréia. A influência de Lactobacillus casei Shirota sobre a microbiota intestinal foi avaliada em um estudo prospectivo, randomizado, duplo-cego e controlado. Sessenta e três crianças hospitalizadas com idade de 2 a 14 anos, sob tratamento com antibióticos β-lactâmicos, foram randomizadas para receber o leite fermentado por L. casei Shirota, 108-9 UFC/mL, ou o placebo, durante o tratamento antimicrobiano. As amostras de fezes foram colhidas antes da administração do leite fermentado, durante o tratamento antibiótico e uma semana após o término do tratamento com o antimicrobiano e a ingestão do leite fermentado. O número de L. casei Shirota aumentou significativamente (p<0,001) durante o período de ingestão do leite fermentado. Foi observado na microbiota do grupo que recebeu o placebo um aumento na contagem de Pseudomonas aeruginosa (p<0,05) e Clostridium sp (p<0,05), principalmente no último período da terapia antimicrobiana. A alteração da microbiota intestinal em decorrência do tratamento antibiótico foi constatada pela diminuição de acetato (p<0,05), butirato (p<0,05) e formato (p<0,05). Embora nenhuma criança deste estudo tenha apresentado diarréia, na avaliação geral, a microbiota daquelas que receberam o leite fermentado mostrou uma recuperação precoce da microbiota intestinal. Foi observado que a variação da contagem bacteriana realizada não foi significativa para as crianças do grupo que recebeu o leite fermentado, enquanto que no grupo placebo a contagem bacteriana ficou alterada, mostrando desequilíbrio da microbiota. Cerca de 50% das crianças ainda apresentaram L. casei Shirota nas fezes após uma semana da ingestão do leite fermentado. Este estudo mostrou que a ingestão do leite fermentado contendo L. casei Shirota promoveu um reequilíbrio mais rápido da microbiota intestinal quando comparada com a do grupo que ingeriu o placebo.
Antimicrobial treatment can destroy the balance of gastrointestinal microflora, which may induce clinical symptoms, mainly diarrhoea. The influence of Lactobacillus casei Shirota on the intestinal microflora was assessed in a prospective, randomised, double-blind controlled study. Sixty-three hospitalised children, with ages between 2 and 14 years, under treatment with β-lactam antibiotics were randomised to receive milk fermented by L. casei Shirota, 108-9 CFU/mL, or placebo during the antimicrobial treatment. Stool samples were collected before the administration of fermented milk, during the antibiotic treatment, and one week after the end of treatment with the antimicrobial agent and the ingestion of fermented milk. The number of L. casei Shirota increased significantly (p<0.05) during the period in which fermented milk was ingested. An increase in the Pseudomonas aeruginosa (p<0.05) and Clostridium sp (p<0.05) count was observed in the microflora of the group that received placebo, mainly in the last period of antimicrobial therapy. The alteration of intestinal microflora as a result of antibiotic treatment was found by the reduction of acetate (p<0.05), butyrate (p<0.05) and formate (p<0.05). The variation in bacterial count proved not to be significant for the children under antimicrobial treatment who received fermented milk, while the placebo group showed imbalance of microflora with the result of the altered bacterial count. About 50% of the children still presented L. casei Shirota in their stools after interrupting the ingestion of fermented milk for one week. This study showed that ingestion of fermented milk containing L. casei Shirota promoted a much faster re-balance of the intestinal microflora when compared to the group that ingested a placebo.

碩士
國立中興大學
畜產學系
91
Lactic acid bacteria and bifidobacteria were widely used as probiotics in foods. As consumers concern more about health, the fermented milk of probiotics become more popular. Probiotics improve the balance of intestinal flora, which exert beneficial effects by decreasing harmful bacterial metabolites such as amines and indoles. Some probiotic bacteria have been found to enhance the immunity of the host and prevention of infectious diseases. The purposes of this study were to examine the antioxidant activity and immunomodulating effect of lactic acid bacteria and bifidobacteria fermented milk. The strains have previously shown the antioxidant activity, the tolerance of gastric acid and bile salts by whole cells. Eight strains of probiotics were selected for antioxidant activity test. The fermented milk had better antioxidant activity than the centrifugal whey and curd of fermented milk. B. adolescentis CCRC 14607 predented the best antioxidant activity by the inhibition of linoleic acid oxidation and it was equal to 1 mg/ml BHT. B. catenulatum CCRC 14667 was the best for reducing activity and it achieved 68% of 1 mg/ml BHT. L. acidophilus CCRC 10695 showed the commensurate scavenging effects on α-diphenyl-2-picryl hydrazyl (DPPH．) radical as vitamin C. In respect of chelating effect on Fe2+, L. brevis CCRC 12187 was equal to 1 mg/ml EDTA. L. bulgaricus CCRC 14009 and B. bifidum CCRC 14670 both represented the greatest ability of scavenging H2O2. The scavenging effects of L. bulgaricus CCRC 14009 on hydroxyl radicals reached 42%. Comparing the antioxidant activity of the eight strains, we selected B. adolescentis CCRC 14607、B. catenulatum CCRC 14667、L. acidophilus CCRC 10695 and L. brevis CCRC 12187 were selected to test their effect on immunity. In order to study the effect on macrophage activation of probiotics, cytokines, H2O2 and NO which was secreted from mouse macrophage cell line RAW 264.7 were examined. The cells were incubated with latic acid bacteria cells, the spent culture supernatant and the fermented milk. The cells of L. brevis CCRC 12187 were the most powerful stimulus for TNF-α production although the fermented milk of some strains could inhibit it. The cells and the cultured broth of probiotics represented the same activation level on macrophage. The fermented milk of L. brevis CCRC 12187 and B. catenulatum CCRC 14667 showed much lower level than control group. The in vivo test of these four strains were studied. The pregnant mice were fed with the reconstituted fermented milk of probiotics. During the period of administration, there were no significant difference on body weight and IgG level in the blood of mother mice. As the time of administration increased, more fecal IgA were indentified. B. adolescentis CCRC 14607 was the most significant strain. All the dams of these mice had higher IgA level than the control group.

碩士
國立中興大學
食品暨應用生物科技學系所
103
Hyptis suaveolens is used as a common traditional herb in Taiwan and also Latin America. It provides a lot of benefits for our health, such as antioxidant, anti-inflammatory, antibacterial, anti-tumor, wound healing, etc. The seeds of Hyptis suaveolens can swell and form a mucus-like polysaccharide on the surface of seeds when soaked in water. Studies indicated that plant polysaccharides can be used as dietary fiber and prebiotics. So far, the research of Hyptis suaveolens is mainly focused on the essential oils, which extracted from the stem and leaves of the Hyptis suaveolens. In this study, we are focused on the polysaccharides of Hyptis suaveolens seeds extracted by water and evaluating its antioxidant capacity, anti-bacterial ability, the potential as a prebiotic to enhance the survival rate of the lactic acid bacteria Streptococcus thermophilus BCRC 13680, Lactobacillus delbrueckii subsp. Bulgaricus BCRC 10696 and Lactobacillus paracasei BCRC 12188 in MRS broth and in fermented milk during cold storage, and the viability of the lactic acid bacteria in the gastrointestinal tract. In our results showed that the Hyptis suaveolens seed polysaccharide had good antioxidant capacity, particularly the polysaccharide extracted at 100℃. At the concentration of 2 mg/mL, the total antioxidant capacity was 96.39%, the ferrous ion chelating ability was 58.15%, the DPPH scavenging ability was 55.61%, and the reducing power was 0.68 which was equivalent to the absorbance of 0.0813 mg/ml ascorbic acid. Supplement with Hyptis suaveolens seed polysaccharides enhanced survival of lactic acid bacteria used in this study in MRS broth during cold storage, but the effect varied among the species of bacteria. Overall, supplemented polysaccharides medium concentration (0.5-2% (v/v)) performed better effect to enhance the lactic acid bacteria viability than in lower (0.1% (v/v)) or higher (4 % (v/v)) concentration. The survival rate of lactic acid bacteria with 0.5%(v/v) polysaccharides in stimulated gastric acid (pH 2.0) and bile juice (4.5%, pH 8.0) was higher than control group. Fermented milk with Hyptis suaveolens seed polysaccharides test results show the viability of lactic acid bacteria was relatively better than the results in MRS broth during cold storage. Especially the 0.5% and 1% (v/v) polysaccharide supplement groups, the results were 1.5 to 3.8 times than control group which was without polysaccharide supplement. The results showed Hyptis suaveolens seed polysaccharide could maintain the survival of lactic acid bacteria in fermented milk during cold storage. The sensory evaluation of fermented milk with Hyptis suaveolens seed polysaccharides test showed that most of people with this test had over ordinary degree of preference. In particular, the group with 1% polysaccharide supplement got the best performance in all aspects. This study demonstrated that Hyptis suaveolens seed polysaccharides had good antioxidant capacity, and could enhance the survival of lactic acid bacteria in MRS broth and in fermented milk during the cold storage. Furthermore, most people could accept this kind of fermented milk with Hyptis suaveolens seed polysaccharides. The results in this study suggested that Hyptis suaveolens seed polysaccharide has the potential to be a prebiotic and develop a new functional drink.

D. Tech. Food Technology.
Discusses a range of lactobacilli and bifidobacteria were isolated from functional food products and pharmaceutical preparations and also obtained from culture collections. They were then subjected to phylogenetic analysis for accurate identification and classification and the probiotic properties of the organisms was evaluated. The isolates were then screened for inhibitory activity against a range of pathogenic bacteria and Candida albicans strains. Selected isolates that were found to have the necessary inhibitory and probiotic properties were recommended for inclusion in an envisaged synbiotic, maize-based beverage that would, in a subsequent study, be subjected to a nutritional intervention trial aimed at alleviating oral thrush in human patients. The specific objectives: to investigate and illustrate the superiority of rpoA and pheS gene sequencing compared to 16S rRNA gene sequencing in the identification and phylogenic assignment of Lactobacillus isolates ; to determine the precision of selected protein-coding gene sequencing in comparison with 16S rRNA gene sequencing for the discrimination and phylogenetic analysis of Bifidobacterium isolates ; to investigate the probiotic properties of selected bacterial strains in terms of antibacterial activity, anti-Candida activity, acid resistance, bile tolerance and antibiotic resistance ; to determine the potential of Lactobacillus isolates to inhibit the growth of each of seven Candida albicans strains in fermented maize gruel and to establish the factors contributing to Candida inhibition and to determine the antimicrobial and antioxidant activity of intracellular extracts and to elucidate compounds in methanol extracts from selected Lactobacillus strains.

碩士
國立臺灣海洋大學
食品科學系
102
gamma-Aminobutyric acid (GABA) is a well-known non-protein amino acid, which is widely distributed in animals and plants. GABA has been reported to be a major inhibitory neurotransmitter in the mammalian central nervous system and show antihypertensive activity. This aims of this research are to manufacture a GABA-enriched fermented milk by lactic acid bacteria (LAB) isolated from fish intestines and to evaluate the probiotic potential property of the GABA-producing LAB. Preliminarily, lactic acid bacteria strains were isolated from healthy fish intestine from previous study. Ten lactic acid bacteria strains of 126 LAB strains were screened based on the capacity of synthesizing GABA. And ten GABA-producing strains show that none of them exhibited hemolytic activity. Moreover, five strains exhibited partial bile salt hydrolase activity, including the strain FPS 2520. FPS 2520 also showed a high percentage of adhesion to monolayer of Caco-2 cells. Strain FPS 2520 demonstrated high survival viability to gastrointestinal conditions simulating stomach and duodenum passage. Optimal conditions of strain FPS 2520 for producing GABA in whole milk were: 10% reconstituted whole milk, initial inoculum size of 5 log CFU/mL, addition of 20 mM MSG, 1% brown sugar, and 0.5% yeast extract during fermentation at 37oC for 48 hours. The results indicated FPS 2520 show a probiotic potential property and have a great application in milk fermentation for the production of G (10.67 mg/mL), and the IC50 of ACEI-inhibitory activity is 0.28 ± 0.02 mg/mL. Besides lactic acid bacteria number slightly decreased, pH value, titratable acidity and GABA concentration didn’t have significant difference compare to control at 4oC for 14 days. Using mixed strains, GABA-producing strain FPS 2520 and proteinase-positive FKR 3737 could significantly enhance the GABA production about 14.74 mg/mL.

Probiotics can influence intestinal responses and mucosal immunity either directly or indirectly through transient modulation of the endogenous microenvironment or the immune system. During space travel, astronauts experience various physiological stresses including putting them at risk for infections or inappropriate immune responses. Macrophages and monocytes are a key cell type involved in innate immunity. The effects of dairy milk or soy milk base fermented with S. thermophilus ST5 in combination with either B. longum R0175 or L. helveticus R0052 on the cell line U937 and all-trans retinoic acid differentiated U937 were examined under normal gravity and simulated microgravity conditions, in order to screen for effects on monocytes and macrophages. Soy and milk ferments demonstrated the ability to modulate certain aspects of the innate immune system, both in normal gravity and in simulated microgravity. These probiotics affected U937 cells differently depending on differentiation stage (monocyte or macrophage) and whether or not the cells were tested in regular gravity or in simulated microgravity conditions. These results provide insight into effects on this aspect of innate immunity and may provide guidelines to potential in vivo administration.
UOIT

Submitted in fulfilment of the academic requirements for the Degree in Master's in Food Science and Technology, Durban University of Technology, 2017.
Pigeon pea (Cajanus Cajan) is consumed in many parts of Africa as a source of protein and carbohydrate. It is underutilised and mainly grown for subsistence. Researching on pigeon pea may enhance value addition and increase its utilization. In this study, a non-dairy probiotic yoghurt was prepared from pigeon pea milk. Yoghurt samples were prepared, using 100% pigeon pea milk, pigeon pea/soy milk in the ratio 50:50 and 100% soy bean milk. The yoghurts were inoculated with yoghurt starter cultures and divided into two equal parts. One part inoculated with Propionibacterium freudenreichii was referred to as probiotic yoghurt, while the other part served as the control. The nutritional, sensory and some functional properties of the yoghurt were determined. The microbiological quality of yoghurt samples stored at 4, 10 and 21°C, respectively, for 4 weeks, were monitored and analysed for aerobic spores’ formers, E. coli, total plate counts, mould and Propionibacterium freudenreichii weekly. The protein contents of the yoghurt samples varied from 4.54-5.85% for 100% soymilk and 100% pigeon pea yoghurt respectively. The probiotic yoghurt showed slightly lower protein content than pigeon pea yoghurt alone. All the yoghurt samples had considerably high total solids (16.04-17.41%) and were fairly good sources of amino acids. Essential amino acids in the yoghurt samples were comparable to the FAO/WHO (2007) recommended amino acid requirement for adults. Anti-nutritional factors of yoghurt samples were significantly lower (P≤0.05) than their milk counterparts, which may be attributed to the fermentation process. Probiotic yoghurt samples showed higher firmness than non-probiotic samples. Total plate counts (log 7.01- 7.46 CFU/ml) samples stored for 2 weeks at 4° C were similar. Predominant organisms were LAB and Propionibacterium freudenreichii. Storage temperature of yoghurt samples had an influence on the total plate count and LAB. Total plate count and LAB significant increased approximately by log 2 CFU/ml for the first two weeks of storage. However, moulds and E. coli were not detected in all samples. Beyond 2 weeks of storage, there was significant decline in total plate counts and LAB, while mould grew and increased. Aerobic spore formers and moulds were observed in the control yoghurt. However, E. coli was not found in all yoghurt samples throughout storage period. The pH of the milk in which yoghurt mixtures were formulated, ranged from pH 7 to 6.8 for pigeon pea and soymilk declined significantly as a result of acidification. Decline in pH at 4, 10 and 21°C was significant (p≤0.05) with the rate higher at 21, 10 than 4° C. Decline in pH resulted in increased TTA values over storage temperatures and periods. Samples stored at 21°C and 10°C had significantly higher TTA values than samples stored at 4° C. The colour values evaluated were recorded as L*, b*, a* and ∆E* during 4 weeks storage at 4, 10 and 21° C. Significantly high values (p≤0.05) were recorded for L* yoghurt samples with soymilk. The colour scale defines positive (red) and negative (green) for a* and b* positive (yellow) and negative (blue). All a* values both positive and negative were less than 3. There was no negative value recorded for b*. Colour difference ∆E* values trends increased as storage time and temperature increased. There were significant (p≤0.05) differences between samples stored at same and different storage temperatures and periods. Water holding capacity was significantly different (p≤0.05) in all the yoghurt samples stored at 4, 10 and 21°C for 4 weeks. Formulation with 100% soymilk recorded higher values. Soy yoghurt and probiotic yoghurts (100 %) showed higher water holding capacity compared to pigeon pea yoghurt and pigeon pea/soymilk yoghurt. The addition of Propionibacterium freudenreichii did not significantly affect sensory properties of the yoghurts. Acceptable yoghurt was produced from pigeon pea with comparable quality to soy which serves as control. Proximate composition was comparable to previous reports. Microbial quality and profile of all the yoghurt samples were similar. The absence of pathogenic bacteria in all the yoghurt samples confirm their safety. Soy yoghurt was most acceptable amongst the yoghurt samples but all the samples had comparable ratings, and these ratings are within commercially acceptable range (4 to 9) for yoghurt. Storage at 4oC should be the most acceptable, as storage at 21oC encourage proliferation of contaminant
M

碩士
國立臺灣大學
食品科技研究所
96
Colorectal cancer is a major cause of death from cancer in Taiwan. Approximately 70 % of colorectal cancer is related to dietary habit. Many researches showed that probiotics, both cellular compositions and fermented products have antimutagenic and antitumor activities on mutagens. In present study, we used several probiotics, which have been studied in our laboratory for their acid-resistance, bile salt tolerance, and antimutagenicity against B[a]P, t-BOOH, and H2O2. In addition, MTT assay was used to explore the effects of several probiotics against 4NQO cytotoxicity on Int-407 and the proliferation of human colon cancer cell HT-29 and Caco-2. Furthermore, possible antimutagenic mechanisms of probiotics were investigated by section-preincubation test and comet assay to explore the effects of several probiotics against 4NQO-induced DNA damage on Int-407. The results showed that a high probiotics population inhibits colon cancer cell more effectively than a low probiotics population. Among the strains, the crude cell walls of Bifidobacterium lactis Bb-12 exhibited the most potent inhibition activity in HT-29 and Caco-2 for 48h and the viability were 70.6 % and 63.3 %, respectively. The heat-killed cells of B. lactis Bb-12 and L. casei 01 significantly reduced the genotoxicity of 4NQO-induced DNA damage on Int-407 (p < 0.05). In addition, the crude cell walls and intracellular extracts of probiotics showed inhibitory effects against 4NQO cytotoxicity. The primary mechanism of crude cell walls was desmutagenic effect and intracellular extracts was blocking effect. Besides, fermented milk with B. lactis Bb-12 and L. casei 01 also had inhibition on 4NQO-induced DNA damage and cytotoxicity on Int-407. Their main mechanisms were the same as above, inhibition rate were 37.4 % - 52.5 % and 34.1 % - 41.4 %, respectively.