Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Dairy products form the major part of functional foods available on market place. Fresh cheeses are propitious to addition of probiotics due to their suitable pH, acidity and moisture. The objective of this study was evaluate the potential of cream cheese as a vehicle of probiotics, the physical-chemical, microbiological and sensory characteristics and the lipid profile of this product when additioned of prebiotic and probiotics. Treatments of cream cheeses were produced with addition of different levels of probiotics (Bifidobacterium animalis and Lactobacillus acidophilus) and prebiotic (inulin) in amounts according to Central Composite Design, totalizing 12 treatments, with 4 fatorials, 4 axial, 3 repetitions on central point and 1 control. The characteristics were evaluated in all treatments during storage for 45 days at 8°C. No significant changes were observed on physical-chemical characteristics of fat, protein and ash in comparison with control, while moisture was altered by prebiotic addition. The surface response was done to verify the influence of different levels of inulin and probiotic on pH and tritable acidity during storage; only linear and quadratic terms of probiotic level were significant (p<0,05). The pH and tritable acidity affect the viability of cultures; starter culture (S. termophilus) exhibited high viability during storage. Bifidobacterium animalis showed counts above 106 UFC g-1 during all period, assuring the probiotic potential in all treatments. The counts of Lactobacillus acidophilus reduced along storage, probably by competition with starter culture. Assessment of sensory attributes (global appearance, color, aroma, texture, acidity and taste) was evaluated using test with hedonic scale of 7 levels and ordering test. No significant difference was found by Tukey s test (p>0,05) on assessment using hedonic scale, neither on the ordering test. The linear, quadratic and interaction terms of multiple linear regression were not significant (p>0,05) for any sensory attribute; only the intercept of each equation was significant (p<0,01). Lipid profile was not significantly changed by the different levels of probiotic, with amounts of saturated fatty acids, monounsaturated, polyunsaturated and trans fatty acids similar to that of the milk used on cheese manufacture and control. The different levels of probiotic no altered the concentration of conjugated linoleic acid (CLA) in comparison with control, probably due to the low addition of probiotic, the reduced viability of Lactobacillus acidophilus during storage and the short shelf-life of cream cheese, with small time to produce CLA. In conclusion, cream cheese with addition of prebiotic and probiotic on levels of this study showed functional potential; nevertheless, the amount of probiotic used does not increase the concentration of CLA compared to the traditional product. / Derivados lácteos compõem a maior parte dos alimentos funcionais disponíveis no mercado. Queijos frescos são propícios à adição de probióticos por seu pH, acidez e umidade favoráveis. O objetivo deste estudo foi avaliar o potencial do cream cheese em atuar como condutor de probióticos, as características físico-químicas, microbiológicas e sensoriais e o perfil lipídico do produto quando adicionado de prebiótico e probióticos. As formulações de cream cheese foram elaboradas com adição de diferentes concentrações de probiótico (Bifidobacterium animalis e Lactobacillus acidophilus) e de prebiótico (inulina), em quantidades adotadas de acordo com o Delineamento Central Composto Rotacional, totalizando 12 ensaios, sendo 4 fatoriais, 4 axiais, 3 repetições no ponto central e 1 tratamento-controle. As características foram avaliadas durante o armazenamento do produto por 45 dias a 8°C. Não foi observada mudança significativa nas características físico-químicas quanto a gordura, proteína e cinzas em comparação à formulação-controle, enquanto o teor de umidade foi alterado pela adição do prebiótico. A superfície de resposta foi elaborada para verificar a influência das diferentes concentrações de inulina e probióticos sobre o pH e acidez (expressa em ácido láctico) das formulações durante o armazenamento, tendo somente os termos linear e quadrático da concentração de probióticos apresentado significância (p<0,05). O pH e acidez titulável das formulações influenciaram a viabilidade das culturas, tendo a cultura starter (S.termophilus) alta viabilidade durante todo o armazenamento. Bifidobacterium animalis apresentou contagens acima de 106 UFC/g durante todo o período, garantindo a potencialidade probiótica de todas as formulações. As contagens de Lactobacillus acidophilus declinaram ao longo do armazenamento, provavelmente por competição com a cultura starter. A avaliação dos atributos sensoriais (aparência global, cor, aroma, textura, acidez e sabor) foi realizada através de teste com escala hedônica de 7 níveis e teste de ordenação. Não foi encontrada diferença estatística pelo Teste de Tukey (p>0,05) na avaliação por escala hedônica, tampouco preferência significativa no teste de ordenação. Os termos linear, quadrático e de interação da regressão linear múltipla não foram significativos (p>0,05) para nenhum dos parâmetros sensoriais avaliados; somente o intercepto da equação de cada atributo apresentou significância estatística (p<0,01). O perfil lipídico das formulações não foi significativamente alterado pela adição de diferentes concentrações de probiótico, sendo as quantidades de ácidos graxos saturados, monoinsaturados, poliinsaturados e trans semelhantes aos do leite usado como matériaprima e à formulação-controle. As diferentes concentrações de probiótico igualmente não alteraram a concentração de ácido linoléico conjugado (CLA) em relação ao controle, provavelmente pela reduzida concentração de cultura usada, baixa viabilidade de Lactobacillus acidophilus durante ao armazenamento e ao fato de cream cheese ter vida-de-prateleira breve, com curto espaço de tempo para formação do CLA. Conclui-se que o cream cheese adicionado das concentrações estudadas de pre- e probiótico apresenta potencial funcional; entretanto, a quantidade de probióticos não aumentou o teor de CLA em relação às quantidades do produto tradicional.

This thesis is interested in isolation and identification of probiotic bacteria in three different probiotic products using polymerase chain reaction (PCR). DNA in quality suitable for PCR was isolated from crude lysates using three different types of magnetic microparticles and phenol extraction. Identification genera and species of probiotic bacteria was proven using genus and species specific PCRs. Results were in accordance with data presented by manufacturers.

Background - Irritable bowel syndrome (IBS) is a poorly understood functional gastrointestinal disorder and is a major cause of abdominal discomfort and gut dysfunction. IBS symptoms encompass abdominal pain, bloating, flatulence and irregular bowel movements such as constipation, diarrhoea and alternating bowels, bloating, flatulence and irregular bowel movements. Physiological studies have shown that manipulation of the intestinal microbiota by antibiotics, prebiotics or probiotics can affect intestinal functions in the pathogenesis of IBS. The probiotic concept suggests that supplementation of the intestinal microbiota with the right type and number of live microorganisms can improve gut microbiota composition and promote health in IBS sufferers. Aim - The aim of the main clinical trial is to determine whether ingestion of fermented milk containing Bifidobacterium animalis DN-173010 is associated with improved defecation frequency, stool consistency and quality of life in black South African females with constipation-predominant IBS (IBS-C). Methods - A pilot and process evaluation approach was employed during the current study to examine and understand the feasibility of implementing the study and to explore the facilitating implementation of the main clinical trial. Twenty black female participants, aged 18-60, with IBS-C were recruited from the practices of gastroenterologists, specialist physicians and medical doctors in Soweto. Participants fulfilling the Rome III criteria for IBS-C and inclusion criteria were randomized into two groups to participate in a 4-week, double blind, placebo controlled study. The placebo group received unflavoured sweetened, white base yoghurt and the intervention group received similar yoghurt with the probiotic, Bifidobacterium animalis DN-173010 [>3,4X10⁷ CFU/g]. Participants were required to record their bowel movements daily and IBS symptoms weekly in questionnaires during the four-week study period. Quality of life was assessed at baseline and at the end of the treatment period. Participants visited the study unit weekly to collect the placebo or probiotic study products and return the completed questionnaires during the study period. Results - Seventeen participants completed the study (eight intervention and nine placebo). There were not significant differences in IBS symptoms between the two groups, but differences were observed overtime within groups. The severity of abdominal pain score within both groups was statistically significant (p=0.004), and the number of days with pain was also statistically significant (p=0.00001). The frequency of normal stools reported was statistically significant different compared to all the other stool types (constipation and loose stools) throughout the four-week study period in both the intervention and placebo group. There was no significant difference in the quality of life between the intervention group compared to the placebo group. Conclusion - Process evaluation allows for the monitoring of a programme and corrections of problems as they occur. The intervention is feasible to implement, acceptable and safe to participants. The study indicates that consumption of the probiotic Bifidobacterium animalis DN-173010 for four weeks is not superior to the placebo in relieving IBS symptoms. / MSc (Dietetics), North-West University, Potchefstroom Campus, 2014

Background - Irritable bowel syndrome (IBS) is a poorly understood functional gastrointestinal disorder and is a major cause of abdominal discomfort and gut dysfunction. IBS symptoms encompass abdominal pain, bloating, flatulence and irregular bowel movements such as constipation, diarrhoea and alternating bowels, bloating, flatulence and irregular bowel movements. Physiological studies have shown that manipulation of the intestinal microbiota by antibiotics, prebiotics or probiotics can affect intestinal functions in the pathogenesis of IBS. The probiotic concept suggests that supplementation of the intestinal microbiota with the right type and number of live microorganisms can improve gut microbiota composition and promote health in IBS sufferers. Aim - The aim of the main clinical trial is to determine whether ingestion of fermented milk containing Bifidobacterium animalis DN-173010 is associated with improved defecation frequency, stool consistency and quality of life in black South African females with constipation-predominant IBS (IBS-C). Methods - A pilot and process evaluation approach was employed during the current study to examine and understand the feasibility of implementing the study and to explore the facilitating implementation of the main clinical trial. Twenty black female participants, aged 18-60, with IBS-C were recruited from the practices of gastroenterologists, specialist physicians and medical doctors in Soweto. Participants fulfilling the Rome III criteria for IBS-C and inclusion criteria were randomized into two groups to participate in a 4-week, double blind, placebo controlled study. The placebo group received unflavoured sweetened, white base yoghurt and the intervention group received similar yoghurt with the probiotic, Bifidobacterium animalis DN-173010 [>3,4X10⁷ CFU/g]. Participants were required to record their bowel movements daily and IBS symptoms weekly in questionnaires during the four-week study period. Quality of life was assessed at baseline and at the end of the treatment period. Participants visited the study unit weekly to collect the placebo or probiotic study products and return the completed questionnaires during the study period. Results - Seventeen participants completed the study (eight intervention and nine placebo). There were not significant differences in IBS symptoms between the two groups, but differences were observed overtime within groups. The severity of abdominal pain score within both groups was statistically significant (p=0.004), and the number of days with pain was also statistically significant (p=0.00001). The frequency of normal stools reported was statistically significant different compared to all the other stool types (constipation and loose stools) throughout the four-week study period in both the intervention and placebo group. There was no significant difference in the quality of life between the intervention group compared to the placebo group. Conclusion - Process evaluation allows for the monitoring of a programme and corrections of problems as they occur. The intervention is feasible to implement, acceptable and safe to participants. The study indicates that consumption of the probiotic Bifidobacterium animalis DN-173010 for four weeks is not superior to the placebo in relieving IBS symptoms. / MSc (Dietetics), North-West University, Potchefstroom Campus, 2014

BACKGROUND:Widely considered probiotic organisms, Bifidobacteria are common inhabitants of the alimentary tract of animals including insects. Bifidobacteria identified from the honey bee are found in larval guts and throughout the alimentary tract, but attain their greatest abundance in the adult hind gut. To further understand the role of Bifidobacteria in honey bees, we sequenced two strains of Bifidobacterium cultured from different alimentary tract environments and life stages.RESULTS:Reflecting an oxygen-rich niche, both strains possessed catalase, peroxidase, superoxide-dismutase and respiratory chain enzymes indicative of oxidative metabolism. The strains show markedly different carbohydrate processing capabilities, with one possessing auxiliary and key enzymes of the Entner-Doudoroff pathway.CONCLUSIONS:As a result of long term co-evolution, honey bee associated Bifidobacterium may harbor considerable strain diversity reflecting adaptation to a variety of different honey bee microenvironments and hive-mediated vertical transmission between generations.

Orientador: Thais Borges Cesar / Banca: Paula Garcia Chiarello / Banca: Renata Maria Galvão de Campos Cintra / Banca: Anderson Marliere Navarro / Banca: Katia Sivieri / Resumo: Introdução: Aos compostos do suco de laranja 100% puro (SL) são atribuídos várias propriedades biológicas, como atividade antioxidante, anti-hipertensiva, redutora do colesterol e glicemia sanguíneos, e atualmente estudos in vitro têm mostrado que o suco de laranja pode ter ação seletiva sob a microbiota intestinal. Objetivo: Investigar o impacto do consumo diário de suco de laranja sobre a microbiota intestinal e no perfil glicídico e lipídico em mulheres. Métodos: Foram avaliadas 10 mulheres por período de 120 dias, distribuídos em três etapas: (1ª): dieta livre de citrus 30º dia (30 dias); (2ª) ingestão de 300 mL/dia de SL 90º dia (60 dias); (3ª) dieta livre de citrus 120º dia (30 dias). Avaliação dietética e antropometria foram realizadas quinzenalmente durante o período experimental. Foram colhidas amostras de sangue, urina e fezes, no início do estudo e a cada 30 dias, até o final do experimento. Foi realizado: analise dos parâmetros bioquímicos; identificação dos metabólitos urinários por UHPLC-UV-VIS; e análise da composição da microbiota por sequenciamento de gene RNAr 16S. Resultados: A avaliação da composição corporal e a excreção urinária não mostraram alterações durante as etapas experimentais. Os parâmetros bioquímicos mostraram diminuição da glicose (-6,25%), da insulina (-32,7%) e da resistência à insulina (-43,8%), além do colesterol total (-13,9%), LDL (-16%) e triglicerídeos (-30%) com a ingestão de SL (90º dia). Após a parada do consumo de SL (120º dia), o... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: 100% pure orange juice (OJ) compounds are assigned to various biological properties, such as antioxidant, antihypertensive, blood cholesterol lowering and glycemia activity. Currently, in vitro studies have shown that orange juice can present selective action of intestinal microbiota. Objective: To investigate the impact of daily consumption of orange juice on the intestinal microbiota and on the lipid and glucose profile in women. Methods: Ten women were evaluated for 120 days at three differen stages: (1st) citrus-free diet 30th day (30 days); (2nd) intake of 300 mL / day SL 90th day (60 days); (3rd) citrus-free diet 120th day (30 days). Dietary assessment and anthropometry were performed biweekly during the experimental stages. Samples of blood, urine and feces were taken at the beginning of the study, and every 30 days until the end of the experiment. It was carried out: analysis of the biochemical parameters; identification of urinary metabolites by UHPLC-UV-VIS; and analysis of the composition of the microbiota by 16S rRNA gene sequencing. Results: Changes during the experimental stages were not found in evaluation of body composition and urinary excretion. Biochemical parameters showed a decrease in glucose (-6.25%), insulin (-32.7%) and insulin resistance (-43.8%), total cholesterol (-13.9%), LDL (-16%) and triglycerides (-30%) with ingestion of OJ (90th day). After stopping OJ intake (120th day), the biochemical parameters returned to the initial values... (Complete abstract click electronic access below) / Doutor

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.

Orientador: Katia Sivieri / Orientador no exterior: Lene Jespersen / Banca: Suzana Marta Isay Saad / Banca: Adriane Elisabete Antunes de Moraes / Banca: Carla Raquel Fontana Mendonça / Banca: Marcia Pinto Alves Mayer / Resumo: Diversos subprodutos de frutas tropicais, os quais são frequentemente descartados pelas indústrias alimentícias, apresentam elevado conteúdo de fibras e de compostos bioativos. Estes compostos, assim como determinadas cepas probióticas e algumas pectinas presentes nos subprodutos, têm o potencial de modular a microbiota intestinal humana, promovendo diversos benefícios à saúde, tais como a atenuação de parâmetros relacionados à obesidade. O objetivo deste trabalho foi avaliar os efeitos da pectina do limão, de subprodutos secos de frutas tropicais (acerola e camu-camu) e de diferentes cepas probióticas (Bifidobacterium longum BB-46, Lactobacillus acidophilus LA-5 and L. paracasei L-431) na microbiota intestinal de indivíduos eutróficos e obesos utilizando o Simulador do Ecossistema Microbiano Humano (SEMH®). Seis artigos foram desenvolvidos a fim de se responder os objetivos propostos. O primeiro artigo trata-se de uma mini-revisão e, os cinco restantes, artigos originais. No primeiro artigo, sumarizou-se os principais achados sobre a composição da microbiota intestinal de obesos e, revisou-se as novas estratégias de modulação da microbiota intestinal em favor do tratamento da obesidade. Foi possível mostrar que a composição da microbiota intestinal é essencial para o entendimento de mecanismos envolvidos na etiologia da obesidade e que, várias estratégias, tais como, o consumo de prebióticos e probióticos, bem como a prática de atividade física moderada e regular, podem modu... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Several by-products of tropical fruits, which are often discarded by the food industry, have high fibre content and bioactive compounds. These compounds, as well as certain probiotic strains and some pectins present in the by-products, have the potential to modulate the human gut microbiota, promoting several health benefits, including the attenuation of obesity parameters. The aim of this work was to evaluate the effects of lemon pectin, dried by-products of tropical fruits (acerola and camu-camu), as well as of different probiotic strains (Bifidobacterium longum BB-46, Lactobacillus acidophilus LA-5 and L. paracasei L-431) on the gut microbiota from healthy-weight and obese individuals using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME®). Six articles were developed in order to meet the proposed aims. The first article is a mini-review and the other five are original articles. In the first article, we summarized the principal findings on obesity-related microbiota composition and reviewed new strategies for gut microbiota modulation in favour of obesity treatment. We showed that the composition of the microbiota is essential for understanding the mechanisms involved in the aetiology of obesity and, that several strategies, such as consumption of probiotics and prebiotics, as well as moderate and regular physical activity, can modulate the gut microbiota in favour of obesity treatment. In the second article, the chemical composition, the total phenolic co... (Complete abstract click electronic access below) / Resumé: Flere biprodukter fra tropiske frugter, som ofte kasseres af fødevareindustrien, har højt fiberindhold og bioaktive forbindelser. Disse forbindelser, såvel som visse probiotiske stammer og nogle pektiner, der er til stede i biprodukterne, har potentialet til at modulere den humane tarmmikrobiota, der fremmer flere sundhedsmæssige fordele, herunder dæmpning af fedmeparametre. Formålet med dette arbejde var at evaluere virkningerne af citronpektin, tørrede biprodukter fra tropiske frugter (acerola og camu-camu), samt forskellige probiotiske stammer (Bifidobacterium longum BB-46, Lactobacillus acidophilus LA-5 og L. paracasei L-431) på tarmmikrobioten fra raske og overvægtige individer, under anvendelsen af den humane intestinale mikrobielle økosystem simulator (SHIME®). Seks artikler blev udviklet for at opfylde de foreslåede mål. Den første artikel er en mini-anmeldelse, og de andre fem er originale artikler. I den første artikel opsummerede vi de vigtigste fund om fedme-relateret mikrobiota sammensætning og gennemgik nye strategier for tarmmikrobiota modulering til fordel for fedmebehandling. Vi viste, at mikrobiotas sammensætning er afgørende for forståelsen af mekanismerne i fedmens etiologi, og at flere strategier, såsom forbrug af probiotika og præbiotika, samt moderat og regelmæssig fysisk aktivitet kan modulere tarmmikrobiotaten til fordel for fedme behandling. I det anden artikel blev den kemiske sammensætning, de samlede phenolforbindelser og in vitro antioxidantkapac / Doutor

In this study, strains of probiotic bacteria have been selected for tolerance to low pH, bile, sucrose, oxygen in media and low storage temperatures. Lactobacillus acidophilus 2401 and Bifidobacterium infantis 1912 were selected as strains able to survive in these conditions. These two strains were then offered further protection from the adverse conditions of food processing and storage by microencapsulation in a calcium alginate and starch gel matrix. Encapsulation in calcium alginate increases survival in yoghurt. In cheddar cheese the free L. acidophilus 2401 and B. infantis 1912 cells survived better than the encapsulated cells, probably due to the dense nature of the cheddar cheese matrix combined with the encapsulation restricting the flow of the nutrients and metabolites between the outside environment and the cells. In ice cream survival was high, probably due to the high fat and solids nature of the ice cream combined with the low storage temperature. The trial results of the laboratory scale production was consistent with the survival results for yoghurt and cheddar cheese. Incorporation of encapsulated probiotic bacteria into ice cream and cheddar cheese was acceptable by sensory standards and largely unnoticeable in comparison with the same foods without capsules. The capsules were visible and able to be felt on the tongue when eaten in yoghurt causing the product to be disliked by the panellists. / Master of Science (Hons)

Probiotics and prebiotic are becoming increasingly important to consumers to alleviate issues surrounding gut health, despite the lack of definitive efficacy studies to support health claims. The addition of both probiotics and prebiotics to foods is challenging due to the harsh environmental conditions within the food itself and during transit through the gastrointestinal (GI) tract. To circumvent these challenges encapsulation technology is being explored to protect sensitive ingredients and to control their release within the lower intestines thereby maximizing the health benefiting effects. The overall goal of this research was to design a protein delivery capsule using phase separated pea protein isolate (PPI)-alginate (AL) mixtures for the entrapment of the synbiot which includes the probiotics, Bifidobacterium adolescentis, and the prebiotic, fructooligosaccharides (FOS), such that the capsule design provides highly effective protection and release within the GI tract. Research was carried out in three studies.<p> In study 1, PPIn (native isolate) and AL interactions were studied in dilute aqueous solutions as a function of pH and biopolymer mixing ratio. Turbidimetric analysis and electrophoretic mobility during an acid titration was used to determine conditions where phase separation occurred. Critical structure forming events associated with the formation of soluble and insoluble complexes in a 1:1 PPIn-AL mixture were found to occur at pH 5.00 and 2.98, respectively, with optimal interactions occurring at pH 2.10. As the PPIn-AL ratio increased, critical pH values shifted towards higher pH until a mixing ratio between 4:1 and 8:1was reached, above which structure formation became independent of the ratios through to ratios of 20:1. Electrophoretic mobility measurements showed a similar trend, where the isoelectric point (pI) shifted from pH 4.00 (homogeneous PPIn) to pH 1.55 (1:1 PPIn-AL). As the ratio increased towards 8:1 PPIn-AL, net neutrality values shifted to higher pHs (~3.80) before becoming constant at higher ratios. Maximum coacervate formation occurred at a mixing ratio of 4:1. Based on these findings, capsule design by segregative phase separation was only used in future studies, due to the acidic nature associated with associative phase separation.<p> In study 2, capsule formation using a native and commercial PPI was studied, and showed no difference between the two formulations during challenge experiments in simulated gastric juice (SGJ). As a result study 3 focused on optimization and characterization of capsules prepared using the commercial PPI. Capsule designs were investigated as a function of protein concentration, prebiotic level, and extrusion conditions (20 vs. 27 G needle) in order to determine protective ability for B. adolescentis within SGJ. Capsule designs were also measured in terms of protein and prebiotic retention during the encapsulation process, geometric mean diameter and size distribution, swelling behaviour and release characteristics within simulated intestinal fluids (SIF). All capsules provided adequate protection over the 2 h duration within SGJ. Capsule breakdown and release was similar for all designs within SIF, with a release mechanism believed to be tied to enzymatic degradation of the PPI material within the wall matrix and/or the amount of excessive Na+ present in the SIF. Capsule size was found to be dependent only on the needle gauge used in the extrusion process. Swelling behaviour of the capsules with SGJ was also found to be dependent only on the protein concentration, where capsules shrank once immersed in SGJ.<p> A 2.0% PPI-0.5% AL capsule without FOS and extruded through a 20 G needle represents the best and most cost effective design for entrapping, protecting and delivering probiotic bacteria. Future work to establish the role FOS could play post-release as the entrapping probiotics colonize the GI tract, and the protective effect of the capsules wall on FOS structure during transit is recommended.