Bacteria, belonging to the Lactobacillus bifidus group, were isolated from rectal swabs obtained from healthy babies receiving breast milk. The fermentation properties of these bacteria in cow's milk were studied. Strains potentially suitable for fermentation in a food product were selected according to their ability to produce lactic acid and no gas from lactose, as well as adhering to characteristics common to the bifidus group (Gram positive, catalase negative and blanching morphology). The five strains chosen for intensive study produced 1.14 to 1.25% lactic acid and less than 0.05% volatile acid. The latter organisms could be cultured readily and coagulated milk in less than 24 hr when using a 5% inoculum. Repeated subculture resulted in a change from the branched to the unbranched bacterial morphology. This transformation, which is not unusual among bifid bacteria, was accompanied by a change in ability to produce lactic acid, but no gas was produced by branched or unbranched strains. A great deal of variability in morphology, growth consistency, and acid production was observed among the more than 500 bacterial strains isolated. However, the bifid populations from Caucasian babies was not noticeably different from those of native Indian babies. All of the five strains selected utilized inulin, dextrin, mannitol, and melezitose. A 24 hr culture of fermented milk yielded a product with a mild pleasant natural flavour which could conceivably be altered, if desired, by adding sweeteners or flavouring agents / Land and Food Systems, Faculty of / Graduate

Bifidobacterium bifidum MIMBb75 is a novel strain whose behavior within the intestinal tract is not known. This work attempts to determine the survival and molecular determinant responsible for its colonization through two studies involving administration to mice of 1) B. bifidum MIMBb75 or 2) engineered B. longum NCC2705 expressing the B. bifidum-specific surface protein BopA. MIMBb75 was able to transiently colonize the murine intestinal tract. As a result, endogenous bifidobacteria increased significantly in the proximal colon whereas Clostridia clusters were differentially affected in a region-dependent fashion. Genetically engineered B. longum NCC2705, expressing the BopA gene, did not impact endogenous bacterial groups in the proximal colon or feces, albeit, it down-regulated the gene expression of KC and mucin 4 in the proximal colon. These outcomes indicate that MIMBb75 encompasses characteristics of a probiotic, but further studies are required to assess the role of BopA as the molecular determinant of its action.

Bifidobacterium bifidum MIMBb75 is a novel strain whose behavior within the intestinal tract is not known. This work attempts to determine the survival and molecular determinant responsible for its colonization through two studies involving administration to mice of 1) B. bifidum MIMBb75 or 2) engineered B. longum NCC2705 expressing the B. bifidum-specific surface protein BopA. MIMBb75 was able to transiently colonize the murine intestinal tract. As a result, endogenous bifidobacteria increased significantly in the proximal colon whereas Clostridia clusters were differentially affected in a region-dependent fashion. Genetically engineered B. longum NCC2705, expressing the BopA gene, did not impact endogenous bacterial groups in the proximal colon or feces, albeit, it down-regulated the gene expression of KC and mucin 4 in the proximal colon. These outcomes indicate that MIMBb75 encompasses characteristics of a probiotic, but further studies are required to assess the role of BopA as the molecular determinant of its action.

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Submitted by Lucelia Lucena (lucelia.lucena@ufpe.br) on 2015-03-10T18:06:55Z No. of bitstreams: 2 Gabriela Teixeira _ Dissertação Mestrado.pdf: 1514043 bytes, checksum: f74fa7770b03bc783658b45328e64e05 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-10T18:06:55Z (GMT). No. of bitstreams: 2 Gabriela Teixeira _ Dissertação Mestrado.pdf: 1514043 bytes, checksum: f74fa7770b03bc783658b45328e64e05 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2012-03 / A adição de culturas probióticas tem sido testada em vários produtos lácteos, e a maior parte dos microrganismos consegue permanecer viável, alcançando características desejáveis no produto final. O objetivo da pesquisa foi desenvolver requeijão cremoso com probióticos, Lactobacillus acidophilus e Bifidobacterium bifidum, a fim de se tornar uma alternativa saudável de um alimento de amplo consumo. Foram elaboradas 3 formulações: requeijão cremoso fresco padrão, requeijão cremoso fresco com probióticos, requeijão cremoso fresco probiótico com reduzido teor de gordura. Para a caracterização do produto foram realizadas análises para determinação da composição centesimal, em triplicata para determinação de umidade, cinzas, lipídios e proteínas, e carboidratos. A contagem de coliformes totais e Escherichia coli foram realizadas nas amostras para análise da qualidade higiênico-sanitária. Realizou-se a contagem de L. acidophilus e B. bifidum durante o período de armazenamento de 15 dias. A avaliação da resistência das culturas probióticas às condições digestivas foi realizada empregando-se um modelo in vitro, utilizando suco gástrico e entérico simulados e enzimas do trato gastrintestinal. A análise da cor foi realizada por colorimetria em escala CIELab. A análise sensorial foi aplicada pelo teste de aceitação por escala hedônica para avaliação dos atributos e avaliação da qualidade global e intenção de compra. A composição centesimal dos requeijões padrão e adicionado de probióticos atendeu aos requisitos da legislação para o produto. O requeijão proposto como light apresentou redução de 26% no valor calórico total e 43% no teor de lipídeos em relação ao padrão. Quanto a acidez titulável, observou-se que o armazenamento influenciou significativamente os seus valores nas preparações com probióticos, diferindo do padrão que não apresentou alterações nos valores durante o armazenamento. A qualidade higênico-sanitária foi satisfatória, visto que as amostras foram negativas para presença de coliformes totais e Escherichia coli. As contagens individuais de Lactobacillus acidophilus e Bifidobacterium bifidum, estiveram de acordo com as exigências da legislação para alimentos com alegações de propriedades funcionais. O requeijão cremoso fresco probiótico apresentou contagem de L. acidophilus de 4,0x107UFC/g e B. bifidum de 2,0x108UFC/g aos sete dias de armazenamento e o requeijão cremoso fresco probiótico com reduzido teor de gordura apresentou contagem de L. acidophilus de 5,0x109UFC/g e B. bifidum de 2,0x109UFC/g. A redução de gordura no produto até os níveis estudados não interferiu na viabilidade dos probióticos no requeijão cremoso durante o armazenamento de 15 dias. Após a simulação da digestão in vitro, houve redução da população total de probióticos em ambas as formulações com resultados mais expressivos para a formulação com reduzido teor de gordura afirmando o efeito protetor das gorduras para a sobrevivência dos probióticos ao trato gastrintestinal. A análise instrumental da cor dos requeijões cremosos indicou produtos com boa luminosidade, com acentuação da cor amarela ao longo do tempo de armazenamento. A avaliação sensorial demonstrou que a adição de probióticos resultaram em requeijões cremosos com melhor perfil de características e maior aceitabilidade e intenção de compra que a preparação padrão. Pode-se concluir que os probióticos melhoraram a qualidade sensorial do produto, sendo tais amostras preferidas pelos julgadores. O requeijão cremoso tradicional e com baixo teor de gordura são tecnologicamente viáveis para a incorporação das culturas probióticas de L. acidophilus e B. bifidum.

Made available in DSpace on 2017-07-10T17:48:20Z (GMT). No. of bitstreams: 1 Maikel_Possamai.pdf: 915835 bytes, checksum: b9f7340ba0b838b5bd95229abaed0be6 (MD5) Previous issue date: 2010-08-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Two experiments were conducted to evaluate the effect of using probiotics and symbiotic in piglets diets in the nursery phase. The first experiment was the performance of piglets from 21 to 35, from 36 to 49 days and in total period, being also held the count of lactic bacteria, fecal coliforms and clostridia in the piglets faeces at 35 and 49 days old. The economic viability of probiotics and symbiotic used in piglets diets was also evaluated. There were used 120 piglets, with 21 days old, with initial average weight of 5.80 ± 0.30 kg, distributed in a randomized blocks design, in a factorial scheme consisting of two levels of probiotics (0.30 and 0.60%) and three levels of inulin inclusion (0.00; 0.25 and 0.50%), totalizing six treatments with five replicates. In the metabolism trial were used 24 barrows, with average initial weight of 18.00 ± 0.38 kg, distributed individualy in metabolic cages, and the experimental design and treatments used were the same described in the performance experiment, with four replicates. In this trial were determined the values of digestible energy, metabolizable energy, the digestibility and metabolizability coefficients of gross energy and at the end of metabolism trial, were collected fecal samples, by means of retal massage to determine fecal pH. The inclusion of probiotic and inulin in the diets did not influence (P>0.05) the performance of piglets, from 21 to 35 and from 21 to 49 days of age, the cost index, the index of economic efficiency, the count of lactic bacteria, fecal coliforms and clostridia in the faeces. In the metabolism trial was observed that the inclusion of probiotics in the diets did not influence in the energy metabolization. Nevertheless, the levels of inclusion of inulin reduced (P<0.05) the digestible energy and the digestibility coefficient of gross energy, but did not influence in the coefficient of gross energy metabolization. The pH fecal values were not influenced by the inclusion of probiotic and symbiotic. The use of probiotic and inulin, and their association, did not change the performance, thefaeces microbial counts, the economic viability and gross energy metabolization of piglets from 21 to 49 days old / Foram realizados dois experimentos para avaliar o efeito do uso de probióticos e simbiótico em rações para leitões na fase de creche. O primeiro experimento foi o de desempenho dos leitões dos 21 aos 35, dos 36 aos 49 dias e no período total, sendo também realizada a contagem das bactérias lácticas, coliformes e clostrídios nas fezes dos leitões, aos 35 e 49 dias de idade. A viabilidade econômica da utilização dos probióticos e simbiótico nas rações dos leitões também foi avaliada. Foram utilizados 120 leitões, com 21 dias de idade, com peso médio inicial de 5,80 ± 0,30 kg, distribuídos em um delineamento experimental de blocos ao acaso, em um esquema fatorial constituído de dois níveis de probióticos (0,30 e 0,60%) e três níveis de inclusão de inulina (0,00; 0,25 e 0,50%), totalizando seis tratamentos com cinco repetições. No ensaio de metabolismo foram utilizados 24 suínos machos castrados, com peso vivo médio inicial de 18,00 ± 0,38 kg, distribuídos individualmente em gaiolas de metabolismo, e o delineamento experimental e tratamentos utilizados foram os mesmos descritos no experimento de desempenho, com quatro repetições. Foram determinados os valores de energia digestível, energia metabolizável, os coeficientes de digestibilidade e metabolizibilidade da energia bruta e no final do ensaio metabólico foram coletadas amostras de fezes, por meio de massagem retal, para determinar o pH fecal. A inclusão do probiótico e inulina nas rações não influenciou (P>0,05) o desempenho dos leitões, dos 21 aos 35 e dos 21 aos 49 dias de idade, o índice de custo, o índice de eficiência econômica, a contagem de bactérias acidoláticas, coliformes e clostrídios das fezes. No ensaio metabólico foi observado que a inclusão de probiótico nas rações não influenciou na metabolizibilidade da energia. No entanto, os níveis de inclusão de inulina reduziram (P<0,05) os valores de energia digestível e o coeficiente de digestibilidade da energia bruta, mas não influenciaram no coeficiente de metabolizibilidade da energia bruta. Os valores de pH fecal não foram influenciados pela inclusão de probiótico e simbiótico. O uso de probiótico e inulina, e sua associação, não alteraram o desempenho, a contagem de micro-organismos, a viabilidade econômica e a metabolizibilidade da energia bruta de leitões dos 21 aos 49 dias de idade

Made available in DSpace on 2017-07-10T17:48:24Z (GMT). No. of bitstreams: 1 Wagner_Thiago_Mozer_da_Silva.pdf: 197351 bytes, checksum: b4b84a219d2e39d9b19bb5c957000af1 (MD5) Previous issue date: 2010-08-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this work was evaluate the effect of different levels of inclusion of inulin (chicory root extract) in diets containing ingredients of plant and animal on performance, carcass characteristics (cuts and abdominal fat) and the parameters blood, for broiler chickens 10-40 days old. 1056 chicks were used from one day old, housed on reused litter in a completely randomized design with eight treatments and six replicates of 22 birds each, which were arranged in a factorial 2 x 4, where the first factor was the use of two diets (plant and animal) and the second factor represented by four levels of inulin (0.00, 0.25, 0.50, 0.75%). The probiotic used was composed of Lactobacillus acidophillus, Bifidobacterium bifidum and Streptococcus faecium. The characteristics evaluated were weight gain, final weight, average feed intake, feed conversion, viability 1-40 days of age. At 21 and 35 days of age were measured blood parameters (calcium, phosphorus, uric acid and total protein). At 40 days was carried out the slaughter of birds to assess the yield of carcass, breast, drumstick, thigh, wing and abdominal fat percentage. Within the analysis of economic viability was calculated the cost of the experimental diets, the index of economic efficiency and cost index. In the initial phase and growth was not found significant results for the performance variables. In the total periods of 1-40 days of age for the variable feed no significant effect (P <0.05) for levels of inclusion. Carcass yield was influenced (P <0.05) by food type regardless of the levels of inclusion of inulin. For the cuts, only the thigh showed a significant difference (P <0.05) for the type of diet. Was observed in abdominal fat interaction (P <0.05) between type of diet (plant and animal) and the levels of inclusion of inulin. For the blood parameters for 21 days was significantly different (P <0.05) between levels of inulin inclusion for the variables P and total protein, and 35 was again a significant difference (P <0.05) for phosphorus . The other variables were not affected with the use of inulin. The use of 2.5 g / kg inulin provides better feed for broilers at 1-40 days old. Inclusion of inulin reduces abdominal fat independent mind the type of diet. The use of feed containing animal ingredients provides improved higher carcass yield and higher thigh yield. The use of inulin affected the phosphorus stock you live at 21 and 35 days old. The use of inulin in diets of broiler feed cost rises mind independent of the use of ingredients of animal or vegetable / Objetivou-se com este trabalho avaliar o efeito de diferentes níveis de inclusão de inulina (extrato de raiz chicória) em rações contendo ingredientes de origem vegetal e animal, sobre o desempenho, as características de carcaça (cortes nobres e gordura abdominal) e os parâmetros sanguíneos, para frangos de corte de 1 a 40 dias de idade. Foram utilizados 1056 pintos de um dia de idade, alojados sobre cama reutilizada, em um delineamento experimental inteiramente casualizado, com oito tratamentos e seis repetições e 22 aves por unidade experimental, os quais foram arranjados em um esquema fatorial 2 x 4, onde o primeiro fator foi a utilização de duas dietas (vegetal e animal) e o segundo fator representado por quatro níveis de inclusão de inulina (0,00; 0,25; 0,50; 0,75%). O probiótico utilizado era composto por Lactobacillus acidophillus; Streptococcus faecium e Bifidobacterium bifidum. As características avaliadas foram ganho de peso, peso final, consumo médio de ração, conversão alimentar, viabilidade de 1 a 40 dias de idade. Aos 21 e 35 dias de idade foram mensurados os parâmetros sanguíneos (cálcio, fósforo, ácido úrico e proteínas totais). Aos 40 dias foi realizado o abate das aves para avaliar o rendimento de carcaça, peito, coxa, sobrecoxa, asa e percentual de gordura abdominal. Dentro da análise da viabilidade econômica foram calculados os custos das dietas experimentais, o índice de eficiência econômica e o índice de custo. Na fase inicial e de crescimento não foi encontrado resultado significativo para as variáveis de desempenho. No período total de criação de 1 a 40 dias de idade, para a variável conversão alimentar houve efeito significativo (P<0,05) para os níveis de inclusão. O rendimento de carcaça foi influenciado (P<0,05) pelo tipo de ração independente dos níveis de inclusão de inulina. Para os cortes, somente a coxa apresentou diferença significativa (P<0,05) para o tipo de ração. Na gordura abdominal foi observado interação (P<0,05) entre o tipo de ração (vegetal e animal) e os níveis de inclusão de inulina. Para os parâmetros sanguíneos aos 21 dias foi observada diferença significativa (P<0,05) entre os níveis de inclusão inulina para as variáveis fósforo e proteínas totais, e aos 35 novamente foi observada diferença significativa (P<0,05) para o fósforo. As demais variáveis não foram afetadas com a utilização da inulina. A utilização de 2,5g/kg de inulina proporciona melhor conversão alimentar para aves de 1 a 40 dias de idade. A inclusão de inulina reduz a deposição de gordura abdominal independentemente do tipo de ração. A utilização de ração com ingredientes de origem animal proporciona melhor rendimento de carcaça e maior rendimento de coxa. A utilização de inulina afetou os níveis de fósforo circulante aos 21 e 35 dias de idade. A utilização de inulina em rações de frango de corte eleva o custo da ração independente mente da utilização de ingredientes de origem animal ou vegetal

Lactic acid bacteria have been reported to be used as a health adjunct in food for u many years. However, these health benefits have not been proven. and how these bacteria pass through the digestion process and remain viable in the human intestinal tract is still not clear. The aim of this work was to isolate mutants from Lactobacillus acidophilus or Bifidobacterium bifidum that could tolerate the conditions of the digestion process (low pH and bile conduction) and to characterize these isolated mutants. Acid- and bile-tolerant mutants of L. acidophilus were isolated from parental strains successfully using natural selection techniques. These mutants survived and grew at conditions of pH 3.5 with 0.2% mixed bile salts added. After the selection, phenotypic characterization was identified to further clarify desirable traits for use as probiotic adjuncts in foods. These phenotypic characteristics included protease, aminopeptidase, ß-galactosidase, and bile salt hydrolase activity. Based on different protease, aminopeptidase, and ß-galactosidase activity, selected acid- and bile-tolerant mutants contained different growth characteristics compared with their parents. All the isolates tested showed different bile salt hydrolase activity, and this activity was not strain and medium dependent. Plasmid profiles and fatty acid analysis were conducted to provide more information of these acid/bile tolerant isolates and whether or not they were mutants from their parent strains rather than only adapted variants. Results showed the acid-/bile-tolerant isolates contained different plasmid profiles and cell wall fatty acids compared with their parents, which indicated these isolates were mutants. Protein expression by two-dimensional gel electrophoresis showed different protein expression patterns between acid- and bile-tolerant mutants and their parents. fm1her suggesting these isolates were mutants. We observed the protein production in parent strains decreased as the pH decreased. and protein expression in mutants remained the same as pH decreased. Two of the proposed health benefits of probiotic bacteria are anticholesterol activity and antimicrobial activity. These were evaluated using selected acid- and bile-tolerant mutants. Results showed no decrease of cholesterol in the test medium during bacterial growth. The observed antimicrobial activity was due to the presence of active cells. and this may relate to the acid production during cell growth and not to the production of antimicrobial substances. We concluded that the acid-/bile-tolerant isolates were mutants, and they survived and grew better in harsh environments compared with their parent strains. These mutants may be useful as a food adjunct in the future, but further study is needed to establish their use and possible probiotic benefits.

Ice cream mix (12% fat, 11% milk solids nonfat, 12.5% sugar, and 4.5% corn syrup solids) was fermented with Lactobacillus acidophilus and Bifidobacterium bifidum. Half of the mix was heat treated at 82°C for 30 minutes and cooled to 40-41°C. The other half was warmed to 40-41°C and inoculated with the starter cultures. Both were made into ice cream and stored at -29°C. Survival of L. acidophilus and B. bifidum and of β-galactosidase activity were monitored during 17 weeks of frozen storage. Reinforced clostridial medium was used to enumerate culture bacteria. Colony counts, after fermentation, for both L. acidophilus and B. bifidum were about 5 x 108. The population of cultures decreased less than one log cycle after initial freezing. After 17 weeks storage the bacterial counts were 1 x 107 for B. bifidum and were 4 x 106 for L. acidophilus. During the same period, β-galactosidase activity decreased only 31%. Therefore, frozen fermented dairy products provide a good vehicle to supply β-galactosidase enzymes to people who are lactose maldigestors. Frozen fermented ice cream was prepared at four different pH's (5.0, 5.5, 6.0, 6.5) by blending fermented mix with unfermented mix and then was frozen to produce samples for sensory evaluation. All samples were strawberry flavored. These were then evaluated by 88 judges. The preferred pH, based on overall acceptance, was 5.5. A second sensory evaluation was conducted to compare heat-treated with non-heat-treated ice cream. There were no significant differences in appearance, texture, flavor, and overall acceptance between the two samples. Our study shows that ice cream is a suitable vehicle for delivering these beneficial microorganisms and enzymes to consumers.