Search

Thanks to the beneficial properties of probiotic bacteria, there exists an immense demand for their consumption in probiotic foods worldwide. Nevertheless, it is difficult to retain a high number of viable cells in probiotic food products during their storage and gastrointestinal transit. Microencapsulation of probiotic bacteria is an effective way of enhancing probiotic viability by limiting cell exposure to extreme conditions via the gastrointestinal tract before releasing them into the colon. This research aims to develop a new coating material system of microencapsulation to protect probiotic cells from adverse environmental conditions and improve their recovery rates. Hence, Lactobacillus rhamnosus was encapsulated with emulsion/internal gelation techniques in a calcium chloride solution. Alginate–probiotic microbeads were coated with xanthan gum, gum acacia, sodium caseinate, chitosan, starch, and carrageenan to produce various types of microcapsules. The alginate+xanthan microcapsules exhibited the highest encapsulation efficiency (95.13 ± 0.44%), they were simulated in gastric and intestinal juices at pH 3 during 1, 2, and 3 h incubations at 37 °C. The research findings showed a remarkable improvement in the survival rate of microencapsulated probiotics under simulated gastric conditions of up to 83.6 ± 0.89%. The morphology, size, and shape of the microcapsules were analyzed using a scanning electron microscope. For the protection of probiotic bacteria under simulated intestinal conditions, alginate microbeads coated with xanthan gum played an important role, and exhibited a survival rate of 87.3 ± 0.79%, which was around 38% higher than that of the free cells (49.4 ± 06%). Our research findings indicated that alginate+xanthan gum microcapsules have a significant potential to deliver large numbers of probiotic cells to the intestines, where cells can be released and colonized for the consumer’s benefit.

Various microbial polymers, namely xanthan gum, gellan gum, pullulan gum and jamilan, were tested as a suitable encapsulating material for Lactobacillus plantarum CRL 1815 and Lactobacillus rhamnosus ATCC 53103. Resulting capsules were also studied for their pH and simulated gastrointestinal conditions tolerance. The morphology of the microcapsules was studied using scanning electron microscopy. pH tolerance was tested at pH 2.0, 3.5, 5.0 and 6.5 over a 6. h incubation period. Simulated gastrointestinal conditions were assayed with simulated gastric and pancreatic juices and simulated bile over a 24. h incubation period. Suspensions of probiotic organisms were used as a control. The results from encapsulation with microbial polymers indicate that mixtures of 1% xanthan gum with 0.75% gellan gum and 1% jamilan with 1% gellan gum were the most suitable for microencapsulation. Results for the pH tolerance tests showed no improvement in the viability of cells in relation to the control, except for pH 2.0 where lactobacilli encapsulated in xanthan:gellan gum (1%:0.75%) prolonged their viability by 6. h exposure. Xanthan:gellan gum (1%:0.75%) was the most effective of the encapsulating materials tested in protecting L. plantarum and L. rhamnosus against simulated bile, improving its viability in 1-2. log. CFU when compared with control. The results of this study suggest that microbial polymers are an interesting source of encapsulating material that should be taken into account for prospective studies of probiotic encapsulation for oral delivery applications. Fil: Jiménez Pranteda, María Luján. Universidad de Granada; España. École Nationale Nantes Atlantique Vétérinaire, Agroalimentaire et de l'Alimentation; Francia. Centro de Investigación Biomédica. Instituto de Nutrición y Tecnología de los Alimentos; España Fil: Poncelet, Denis. École Nationale Nantes Atlantique Vétérinaire, Agroalimentaire et de l'Alimentation; Francia Fil: Nader, Maria Elena Fatima. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Arcos, Antonio. Universidad de Granada; España Fil: Aguilera Gómez, Margarita. Universidad de Granada; España. Centro de Investigación Biomédica. Instituto de Nutrición y Tecnología de los Alimentos; España Fil: Monteoliva Sánchez, Mercedes. Universidad de Granada; España. Centro de Investigación Biomédica. Instituto de Nutrición y Tecnología de los Alimentos; España Fil: Ramos Cormenzana, Alberto. Universidad de Granada; España. Centro de Investigación Biomédica. Instituto de Nutrición y Tecnología de los Alimentos; España

Orientador: Mirna Lúcia Gigante Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos Resumo: A microencapsulação de micro-organismos probióticos tem se mostrado como uma técnica promissora para manter sua viabilidade tanto nos produtos aos quais são adicionados, bem como durante sua passagem pelo trato gastrointestinal. O objetivo geral deste trabalho foi avaliar a fabricação e as características durante o armazenamento refrigerado de iogurte batido probiótico adicionado de Lactobacillus acidophilus nas formas livre e encapsulada. O micro-organismo probiótico foi microencapsulado por gelificação iônica e coacervação complexa, utilizando-se pectina e concentrado proteico de soro como materiais de parede. Antes de serem adicionadas ao iogurte, as microcápsulas foram avaliadas quanto à característica físico-química e viabilidade do micro-organismo probiótico. Os iogurtes foram fabricados com leite homogeneizado esterilizado e padronizado adicionado de 2,5% (v/v) da cultura láctica tradicional para fabricação de iogurte e submetidos aos seguintes tratamentos: 1) adição de 1% (v/v) de L. acidophilus livre; 2) adição de 10% (p/v) de microcápsulas úmidas contendo L. acidophilus e 3) adição de 13% (p/v) de microcápsulas liofilizadas reidratadas contendo L. acidophilus. O tempo de fermentação foi considerado o necessário para que o produto atingisse pH 4,8 ± 0,05. Os produtos foram caracterizados quanto à sua composição centesimal e população dos microorganismos. Após 1, 7, 14, 21, 28 e 35 dias de armazenamento refrigerado, os iogurtes foram avaliados quanto à pós-acidificação e viabilidade dos micro-organismos da cultura tradicional de iogurte e probiótica. A viabilidade do L. acidophilus durante a simulação da passagem do iogurte através das condições gastrointestinais foi avaliada após 35 dias. O experimento completo foi repetido três vezes e o delineamento experimental utilizado foi o Split¿plot, em um planejamento do tipo fatorial 3 x 6, em blocos completamente aleatorizados. O efeito dos tratamentos, do tempo de armazenamento bem como da interação desses fatores sobre as variáveis estudadas foi avaliado por ANOVA aplicando-se teste de Tukey para comparação entre as médias ao nível de 5% de significância. O tempo de fermentação dos produtos variou de 180 a 200 minutos. Um dia após a fabricação, o iogurte obtido com o micro-organismo livre apresentou menor pH que os iogurtes adicionados do micro-organismo encapsulado. Os iogurtes produzidos com o micro-organismo encapsulado apresentaram maior teor de proteína quando comparados ao produzido com o L. acidophilus livre. A interação entre os tratamentos e o tempo de armazenamento refrigerado afetou significativamente a pós-acidificação e o número de células viáveis de S. thermophilus, L. bulgaricus e L. acidophilus no iogurte. Os iogurtes probióticos adicionados de L. acidophilus na forma microencapsulada apresentaram menor pós-acidificação e maior sobrevivência do micro-organismo probiótico após 35 dias de armazenamento quando comparado ao iogurte com probiótico adicionado na forma livre. Na forma livre, a população de células viáveis de L. acidophilus reduziu 0,98 ciclos logarítmicos, enquanto na forma encapsulada (úmida e liofilizada) essa redução foi de 0,20 ciclos logarítmicos. Esses resultados indicam a sobrevivência de aproximadamente 10% e 62% do L. acidophilus nas formas livre e encapsulada, respectivamente. Após 35 dias de armazenamento refrigerado, a simulação da passagem dos produtos pelo trato gastrointestinal indicou maior sobrevivência do L. acidophilus encapsulado do que do micro-organismo livre. A microencapsulação do L. acidophilus por gelificação iônica e coacervação complexa utilizando-se pectina e proteína de soro como materiais de parede, conferiu proteção ao micro-organismo, tanto durante o armazenamento refrigerado do produto por 35 dias, como durante a simulação gastrointestinal Abstract: Microencapsulation of probiotic microorganisms has been shown to be a promising technique to maintain their viability either in products to which they are added and during their passage through the gastrointestinal tract. The aim of this study was to evaluate the manufacture and characteristics during refrigerated storage of probiotic stirred yoghurt added of Lactobacillus acidophilus in both free and encapsulated forms. The probiotic microorganism was microencapsulated by ionotropic gelation and complex coacervation, using pectin and whey protein concentrate as wall materials. Before being added to yoghurt, the microcapsules were evaluated for physical chemical characteristics and viability of the probiotic microorganism. The yoghurts were manufactured with sterilized, homogenized and standardized milk added of 2.5% (v/v) yoghurt starter culture and submitted to the following treatments: 1) addition of 1% (v/v) of free L. acidophilus, 2) addition of 10% (w/v) of moist microcapsules containing L. acidophilus and 3) addition of 13% (w/v) of rehydrated freeze-dried microcapsules containing L. acidophilus. The fermentation period was considered the enough time for the product to reach pH 4.8 ± 0.05. The products were characterized to their centesimal composition and population of microorganisms. After 1, 7, 14, 21, 28 and 35 days of refrigerated storage, the yoghurts were evaluated for post-acidification and viability of yoghurt starter culture and probiotic microorganism. The viability of L. acidophilus during simulation of the passage of yoghurt through gastrointestinal conditions was also evaluated after 35 days. The complete experiment was repeated three times and the experimental design was a Split-Plot in a factorial design of the type 3 x 6, in completely randomized blocks. The effect of treatments, storage time and the interaction of these factors on these studied variables was evaluated by ANOVA according to the Tukey test for comparison between the averages, in 5% of significance level. The fermentation period of the products ranged from 180 to 200 minutes. One day after manufacturing, the yoghurt obtained with free microorganism showed lower pH than the yoghurt produced with added encapsulated microorganism. The yoghurts produced with the encapsulated microorganism showed higher protein content than that produced with free L. acidophilus. The interaction between treatments and time of refrigerated storage affected significantly the post-acidification and the number of viable cells of S. thermophilus, L. bulgaricus and L. acidophilus in yoghurt. The probiotics yoghurts added by L. acidophilus in microencapsulated form showed less post-acidification and increased survival of probiotic microorganism after 35 days of storage than yoghurt added of probiotic in free form. The counting of viable cells of L. acidophilus reduced 0.98 log cycles in free form, while in encapsulated form (moist and freeze-dried) the reduction was 0.20 log cycles. These results indicate the survival of approximately 10% and 62% of L. acidophilus in both free and encapsulated forms, respectively. After 35 days of refrigerated storage, the simulation of the passage of yoghurts through the gastrointestinal tract showed a higher survival of L. acidophilus encapsulated than the free microorganism. Microencapsulation of L. acidophilus by ionotropic gelation and complex coacervation, using pectin and whey protein concentrate as wall materials provide protection to microorganism, both during refrigerated storage for 35 days and during simulated gastrointestinal conditions Mestrado Engenharia de Alimentos Mestre em Tecnologia de Alimentos