Dissertations / Theses on the topic 'Synbiotic'

One important development which has recently started to be incorporated to the group of functional foods, is that of the synbiotics (combination of probiotics with prebiotics). This study focused on the development of synbiotic versions of three potential probiotic strains (Lactobacillus acidophilus 74-2, Lactobacillus casei 163 and Bifidobacterium lactis 420) with potential prebiotics and their antipathogenic effects were also investigated. The probiotic characteristics of each of the three strains were assessed prior to investigation of potential synbiotic combinations.

Master of Science
Food Science Institute
Fadi M. Aramouni
Fermented dairy products have long been associated with positive health benefits. Certain probiotics, such as Lactobacillus rhamnosus, have gained popularity as a natural way to improve health and immunity. Fermented dairy products are an easy way to incorporate and deliver probiotics to the consumer. Synbiotic systems are also being investigated as a method for promoting the growth and survival of probiotic bacteria, both during fermentation and after consumption. The purpose of this research was to develop a synbiotic yogurt drink, incorporating L. rhamnosus (probiotic) and inulin (prebiotic), and to investigate the effect of the synbiotic relationship on sensory, physical, chemical, and microbiological properties over time. Yogurt drinks containing the probiotic L. rhamnosus HN001 (treatment P), inulin (treatment I), and a synbiotic system (treatment S) were tested using a randomized block design. Color, viscosity, brix, syneresis, and pH were measured throughout a shelf-life of 28 days. Consumer acceptability was also tested with middle-school aged children. A shelf-life of 28 days at refrigerated temperature was found to be acceptable for the product based on chemical, physical, and microbial analysis. Based on sensory results, the synbiotic drink was found to be acceptable to the target consumers (scoring a 6.82 in “overall liking” on a 9-point hedonic scale), though the treatment with 3% inulin scored slightly higher in most categories (scoring a 7.24 in “overall liking” on a 9-point hedonic scale).

This study examined the possibility of converting mageu, a fermented maize beverage popular throughout southern Africa, into a health-promoting and affordable alternative to probiotic dairy products. A range of probiotic Lactobacillus species was compared with a control species traditionally used to prepare mageu. Prebiotic oligosaccharide (soluble fibre), which enhances the growth of beneficial bacteria, was also included. The resulting beverages were compared in two ways: sensory attributes were determined by a trained panel using quantitative descriptive analysis (QDA), and consumer acceptability was assessed by 53 untrained volunteers. The QDA results suggest that mageu fermented by Lb. acidophilus or Lb. rhamnosus was most similar to the control mageu, while Lb. paracasei mageu and Lb. casei mageu were least similar. The consumer acceptability data confirmed that Lb. acidophilus or Lb. rhamnosus mageu did not differ significantly from the control, suggesting that either of these is suitable for commercial production of probiotic mageu.

L'objectiu d'aquesta Tesi va ser avaluar l'eficàcia de compostos simbiòtics per a millorar la salut i productivitat del bestiar porcí després del deslletament i la seva capacitat per a enfrontar patògens digestius. Per a complir-ho, es van realitzar quatre proves experimentals. En les proves 1 i 2, es van avaluar els efectes d'un simbiòtic format per Bifidobacterium longum subsp. infantis CECT 7210 i inulina enriquida amb oligofructosa en porcs desafiats o no amb Salmonel·la Typhimurium o E. coli enterotoxigènic (ETEC) F4, respectivament. La prova 3 va valorar l'eficàcia d'un probiòtic multisoca compost per la soca anterior i Lactobacillus rhamnosus HN001, barrejats o no amb inulina enriquida amb oligofructosa front a Salmonel·la Typhimurium. La prova 4 va avaluar el mateix probiòtic, administrat o no amb galacto-oligosacàrids en garrins desafiats amb ETEC F4. En les proves 1 i 2 es va seguir un disseny factorial 2x2; tractats o no amb el simbiòtic i desafiats o no amb el patogen. En les proves 3 i 4 es va usar un disseny totalment aleatoritzat constant de cinc grups: un no desafiat (CTR+) i quatre desafiats: la mateixa dieta (CTR-), suplementada amb el probiòtic (PRO), el prebiòtic (PRE) o la combinació dels dos (SYN). Després d'una setmana d'adaptació, els animals van ser desafiats oralment i es va eutanasiar un per corral els dies 4 i 8 (prova 2: 3 i 7) postinfecció. Les principals variables avaluades van ser productivitat, signes clínics, excreció de patogen, perfil fermentatiu, resposta immunitària i morfologia intestinal. El simbiòtic format per Bifidobacterium longum subsp. infantis CECT 7210 i inulina enriquida amb oligofructosa va reduir el número de enterobacteris enganxats i va augmentar la presència de limfòcits intraepitelials en ili de garrins sans. Enfront d'un desafiament, aquest simbiòtic no va poder contrarestar la càrrega de patogen en animals desafiats amb ETEC F4 (prova 2); no obstant, va tendir a accelerar l'eliminació de Salmonel·la (trial 1). Curiosament, en ambdues proves (proves 1 i 2), el simbiòtic va tenir un impacte significatiu en la fermentació colònica, però amb efectes diferents en funció de si els animals havien estat o no desafiats. En la prova 3, la combinació de Bifidobacterium longum subsp. infantis CECT 7210, Lactobacillus rhamnosus HN001 i inulina enriquida amb oligofructosa no va mostrar cap impacte en garrins una setmana després del deslletament. Posteriorment al desafiament, el probiòtic va promoure una eliminació fecal accelerada de Salmonel·la i va millorar la recuperació de l'alteració intestinal. El prebiòtic va portar associat un major nombre de limfòcits intraepitelials. La combinació simbiòtica no va mostrar cap activitat sinèrgica enfront del patogen, perdent la majoria dels efectes esmentats. El simbiòtic avaluat en la prova 4, format per Bifidobacterium longum subsp. infantis CECT 7210, Lactobacillus rhamnosus HN001 i galacto-oligosacàrids, va reduir el número de enterobacteris i coliforms fecals una setmana després del deslletament, en comparació a PRE i PRO. No obstant, després del desafiament amb ETEC F4, el tractament SYN es va associar amb una major càrrega de ETEC F4 en contingut de còlon. Així mateix, les concentracions elevades de TNF-F-α i Pig-MAP van suggerir la presència d'un estat pro-inflamatori sobreestimulat. Per tant, aquesta combinació no va ser capaç de demostrar cap benefici sinèrgic enfront del patogen, perdent alguns dels efectes observats per al probiòtic o el prebiòtic. Els resultats d'aquesta Tesi assenyalen que la combinació de probiòtics i prebiòtics no necessàriament té un efecte additiu o sinèrgic. El seu impacte en la microbiota intestinal i la resposta de l'individu depenen dels desafiaments als quals els animals han d'enfrontar-se. Són necessaris més estudis per a entendre les complexes interaccions que es produeixen en el tracte gastrointestinal i els mecanismes implicats.
El objetivo de esta Tesis fue evaluar la eficacia de compuestos simbióticos para mejorar la salud y productividad del ganado porcino después del destete y su capacidad para enfrentar patógenos digestivos. Para cumplirlo, se realizaron cuatro pruebas experimentales. En las pruebas 1 y 2, se evaluaron los efectos de un simbiótico formado por Bifidobacterium longum subsp. infantis CECT 7210 e inulina enriquecida con oligofructosa en cerdos desafiados o no con Salmonella Typhimurium o E. coli enterotoxigénica (ETEC) F4, respectivamente. La prueba 3 valoró la eficacia de un probiótico multicepa compuesto por la cepa anterior y Lactobacillus rhamnosus HN001, mezclados o no con inulina enriquecida con oligofructosa frente a Salmonella Typhimurium. La prueba 4 evaluó el mismo probiótico multicepa, administrado o no conjuntamente con galacto-oligosacáridos en lechones desafiados con ETEC F4. En las pruebas 1 y 2 se siguió un diseño factorial 2x2; tratados o no con el simbiótico y desafiados o no con el patógeno. En las pruebas 3 y 4 se usó un diseño totalmente aleatorizado constante de cinco grupos: uno no desafiado (CTR+) y cuatro desafiados: la misma dieta (CTR-), suplementada con el probiótico (PRO), el prebiótico (PRE) o la combinación de ambos (SYN). Después de una semana de adaptación, los animales fueron desafiados oralmente y se eutanasió uno por corral los días 4 y 8 (prueba 2: 3 y 7) posinfección. Las principales variables evaluadas fueron productividad, signos clínicos, excreción de patógeno, perfil fermentativo, respuesta inmunitaria y morfología intestinal. La combinación simbiótica formada por Bifidobacterium longum subsp. infantis CECT 7210 e inulina enriquecida con oligofructosa redujo el número de enterobacterias enganchadas y aumentó la presencia de linfocitos intraepiteliales en íleon de lechones sanos. Frente a un desafío, el simbiótico no pudo contrarrestar la carga de patógeno en animales desafiados con ETEC F4 (prueba 2); sin embargo, tendió a acelerar la eliminación de Salmonella (trial 1). Curiosamente, en ambas pruebas (pruebas 1 y 2), el simbiótico tuvo un impacto significativo en la fermentación colónica, pero con efectos diferentes en función de si los animales habían sido o no desafiados. En la prueba 3, la combinación de Bifidobacterium longum subsp. infantis CECT 7210, Lactobacillus rhamnosus HN001 e inulina enriquecida con oligofructosa no mostró ningún impacto en lechones una semana después del destete. Posteriormente al desafío oral, el probiótico promovió una eliminación fecal acelerada de Salmonella y mejoró la recuperación del daño intestinal. El prebiótico trajo asociado un mayor número de linfocitos intrapiteliales. La combinación simbiótica no mostró ninguna actividad sinérgica frente al patógeno, perdiendo la mayoría de los efectos mencionados. El simbiótico evaluado en la prueba 4, formado por Bifidobacterium longum subsp. infantis CECT 7210, Lactobacillus rhamnosus HN001 y galacto-oligosacáridos, redujo el número de enterobacterias y coliformes fecales una semana después del destete de los animales, en comparación a PRE y PRO. No obstante, tras el desafío con ETEC F4, el tratamiento SYN se asoció con una mayor carga de ETEC F4 en contenido de colon. Asimismo, las concentraciones elevadas de TNF-α; y Pig-MAP sugirieron la presencia de un estado proinflamatorio sobreestimulado. Por lo tanto, esta combinación no fue capaz de demostrar ningún beneficio sinérgico frente al patógeno, perdiendo algunos de los efectos observados para el probiótico o el prebiótico. Los resultados de esta Tesis señalan que la combinación de probióticos y prebióticos no necesariamente tiene un efecto aditivo o sinérgico. Su impacto en la microbiota intestinal y la respuesta del individuo dependen de los desafíos a los que los animales deben enfrentarse. Son necesarios más estudios para entender las complejas interacciones que se producen en el tracto gastrointestinal y los mecanismos implicados en ellas.
The main objective of this Thesis was to evaluate the efficacy of different synbiotics to improve health and performance of pigs after weaning, as well as their capacity to fight digestive pathogens. To accomplish it, four experimental trials were performed. In trial 1 and trial 2, the effects of a synbiotic composed by Bifidobacterium longum subsp. infantis CECT 7210 and oligofructose-enriched inulin in pigs challenged or not with Salmonella Typhimurium or enterotoxigenic E. coli F4 (ETEC F4), respectively, were evaluated. Trial 3 assessed the efficacy of a multistrain probiotic composed by the former strain and Lactobacillus rhamnosus HN001, mixed or not with oligofructose-enriched inulin against Salmonella Typhimurium. Lastly, trial 4 evaluated the response of the same multistrain probiotic administered or not with galacto-oligosaccharides in weaned piglets challenged with ETEC F4. A 2x2 factorial design was followed for trials 1 and 2; treated or not with the synbiotic and challenged or not with the pathogen; and, for trials 3 and 4, a completely randomized one, composed by five treatment groups: one non-challenged (CTR+) and four challenged: same diet (CTR-), or supplemented with the probiotic (PRO), prebiotic (PRE) or their combination (SYN). After one week of adaptation, animals were orally challenged and one animal per pen was euthanized at day 4 and 8 (3 and 7 for trial 2) post-inoculation. Main variables assessed were animal performance, clinical signs, pathogen excretion, fermentation profile, immune response and intestinal morphology. The synbiotic combination consisting of Bifidobacterium longum subsp. infantis CECT 7210 and oligofructose-enriched inulin reduced the numbers of ileal attached enterobacteria (trial 2) and enhanced the presence of ileal intraepithelial lymphocytes of healthy weaned pigs. In a challenge, the synbiotic mixture could not counteract pathogen loads in animals challenged with ETEC F4 (trial 2); however, it tended to accelerate Salmonella clearance when it was tested against this pathogen (trial 1). Interestingly, in both trials (trial 1 and trial 2), the synbiotic had a significant impact on the colonic fermentation but with differential effects depending on if animals had been or not challenged. In trial 3, the combination of Bifidobacterium longum subsp. infantis CECT 7210, Lactobacillus rhamnosus HN001 and oligofructose-enriched inulin did not show to have any significant impact in pigs one week after weaning. After the oral challenge, the multistrain probiotic promoted a faster faecal clearance of Salmonella and improved recovery of intestinal damage. Prebiotic treatment increased number of ileal intraepithelial lymphocytes (IEL). The synbiotic combination did not reflect any synergistic activity against the pathogen, losing most of the previous mentioned effects. The synbiotic evaluated in trial 4, composed by Bifidobacterium longum subsp. infantis CECT 7210, Lactobacillus rhamnosus and galacto-oligosaccharides, decreased the number of faecal enterobacteria and coliforms one week after weaning compared to PRE or PRO. Nevertheless, after the ETEC F4 challenge, SYN treatment was associated to the greatest loads of ETEC F4 in colonic digesta. In addition, an enhanced pro-inflammatory status was suggested by the increased serum concentrations of TNF-α and Pig-MAP. This combination was not able, therefore, to demonstrate any synergistic benefits against the pathogen, losing some of the effects found for the probiotic blend or the prebiotic independently. The results exposed in this Thesis highlight that the combination of probiotics and prebiotics not necessarily result in an additive or synergistic effect. Their impact on the intestinal microbiota and the response of the individual probably depend on the challenges that the animals need to face. More studies are needed to understand the complex interactions roduced in the gastrointestinal tract and the involved mechanisms.

Irritable bowel syndrome (IBS) is a condition where abdominal pain, diarrhoea, constipation, gas distension and/or variability in the bowel habit are experienced. It has been suggested that the gut microflora may be involved in this condition and that an 'abnormal' colonic fermentation may provoke the symptoms ofIBS. In this study, the faecal flora of 11. IBS patients was compared with that of 11 healthy individuals quantitatively using fluorescence in situ hylJridisation (FISH) and· traditional plating techniques, as well as qualitatively using denaturating gradient gel electrophoresis (DGGE). It was found that IBS patients had lower total . . ./ bacterial counts, bifidobacteria, lactobacilli, bacteroides and clostridia than seen in healthy individuals. DGGE also showed differences between illS and healthy samples, especially in relation to stability ofthe microflora over time. A synbiotic version ofLactobacillus plantarum NCIMB 41114 with seven different prebiotics was tested in batch culture fermenters using faecal samples from IBS donors. From all the combinations GOS-synbiotic was chosen sirc~ it enhanced the . . growth of the probiotic and was found to be more effective in 'normalising' the . .~ faecal flora in IBS. This synbiotic combination was used in gut model· systems using faecal samples from donors belonging to different IDS subgroups. The results showed that administration of the synbiotic impacted the specific IBS subgroups .. differently, but modulated the rnicroflora towards selective stimulation of bifidobacteria and lactobacilli. In the last part of the study the amount of gas produced from IBS faecal flora during fermentation was compared to that ofhealthy faecal flora. The IDS flora produced more cumulative gas and had greater rates of production than the healthy flora. Administration of the synbiotic reduced gas production in all systems (IBS and healthy). Taken together, these observations help to define dietary management strategies, based on the gut flora and its activities, aimed towards improving IBS symptomology.

A link between the gut and the brain has been proposed to influence psychiatric disorders. Probiotics have been suggested to modify the gut microbiota and thereby improve autism symptoms in children. Attention deficit hyperactivity disorder (ADHD) has high comorbidity with other neuropsychiatric diagnoses, including autism. This is a follow-up of the first study examining a synbiotic intervention in patients with ADHD (Skott et al., 2019). In the original study, 114 adults participated. In this study, 38 adults were evaluated. The aim was to examine if suggested improvements remained 18 months post treatment. Specifically, if reductions were detected in comorbid autism symptoms, emotional dysregulation or functional impairment. The endpoints were measured using questionnaires: Autism-Spectrum Quotient (AQ), Difficulties in Emotion Regulation Scale (DERS-16) and Weiss Functional Impairment Rating Scale (WFIRS). No Synbiotic2000-specific effect was detected. Synbiotic2000 and placebo improved emotion regulation and life skill-functioning equally well. More research is needed to draw reliable conclusions.
En koppling mellan magen och hjärnan har i studier antytts påverka psykiatriska tillstånd. Probiotika har föreslagits förändra mag- och tarmkanalens bakterieflora och därigenom förbättra psykiatriska symtom hos barn med autism. ADHD har hög komorbiditet med andra neuropsykiatriska diagnoser, däribland autism. Detta är en långtidsuppföljning av RCT-studien som var först med att undersöka en synbiotika-intervention hos patienter med ADHD (Skott et al., 2019). I uppföljningen undersöktes 38 av de 114 vuxna som deltagit i originalstudien. Syftet var att undersöka om indikationerna till förbättring höll i sig 18 månader efter studieavslutet. Frågeställningen var om reduktion i komorbida autismsymtom, svårigheter med emotionsreglering eller funktionsnedsättning kunde identifieras. Detta undersöktes genom självskattningsskalor: Autism-Spectrum Quotient (AQ), Difficulties in Emotion Regulation Scale (DERS-16) och Weiss Functional Impairment Rating Scale (WFIRS). Ingen behandlingsspecifik effekt detekterades. Förbättringar av samma grad identifierades av Synbiotic2000 och placebo, utifrån emotionsreglering samt delskalan färdigheter. Mer forskning på området krävs för att möjliggöra tillförlitliga slutsatser.
BAMBA (Behandla Adhd med MagBakterier)

The diseases of the respiratory system occupy the first position which constitutes nearly 62-65 % in the structure of child morbidity. Community-acquired pneumonia (CAP) is one of the most common diseases of modern society. Cytokines play a central role in inflammatory response that is a basis for further immune response. Our aim was to study the effect of synbiotics on pro- and anti-inflammatory serum cytokines (IL-8 and IL-4) concentrations in pre-preschool children with CAP.

Introdução:O envelhecimento está associado a redução na capacidade de lidar/enfrentar estressores, o que significa o principal conceito de fragilidade. Algumas mudanças na composição e função corporal são relacionadas com o desenvolvimento da fragilidade, como: deposição de gordura visceral (altamente relacionada com a inflamação sistêmica) e a redução da gordura subcutânea. Uma alteração importante é a relacionada com um aumento na concentração de marcadores inflamatórios no sangue, o que tem sido associado com um processo mais amplo, denominado inflammaging. Nosso trabalho destaca a hipótese de que esta inflamação de baixo nível é associada a um desbalanço na microbiota e alterações na permeabiliade intestinal, o que então justifica o uso de substâncias simbióticas. Objetivos: avaliar o efeito de uma suplementação por seis meses com substância simbiótica sobre a inflamação sistêmica de idosos em risco de fragilidade. Métodos: foram estudados 49 indivíduos, entre 65-90 anos, que atenderam um ou dois critérios de fragilidade propostos por Fried et al (2001). Tratou-se de um estudo duplo-cego randomizado, no qual os participantes foram alocados em um dos grupos que seguem: SIM (tratamento com simbiótico) - ingestão de uma substância simbiótica (Frutooligossacarideos 6 g, Lactobacillus paracasei 109 a 108 CFU, Lactobacillus rhamnosus 109 a 108 CFU, Lactobacillus acidophilus 109 a 108 CFU e Bifidobacterium lactis 109 a 108 CFU); ou PLA (placebo, controle)- maltodextrina (6g) nas mesmas doses do grupo simbiótico. Ambos os grupos foram instruídos a consumir as substâncias duas vezes ao dia. Antes e após a suplementação os sujeitos foram avaliados para: marcadores plasmáticos de inflamação (IL-6, IL-10 e TNF-α); contagem de células sanguíneas; medidas antropométricas; função intestinal (Escala de Bristol e Critério de ROMAIII). Os dados foram analisados por ANOVA de medidas repetidas (marcadores inflamatórios), com test pos-hoc apropriado, teste T de student (células sanguíneas), bem como odds ratio e redução de risco. Foi adotado o princípio da intenção de tratar. A significância foi considerada para p<0.05. Resultados: O grupo SIM foi composto por 25 sujeitos com idade media de 76.2±8.4 anos, e o grupo PLA foi formado por 24 sujeitos média de idade de 75.6±8.1 anos. A função intestinal melhorou em 5,3% no grupo SIM. Em constraste, o grupo PLA apresentou piora de 20% dos sintomas negativos da função intestinal. O OR da melhora do SIM de acordo com o ROMA III foi de 0.76 (IC= 0.09-6.17), indicando um benefício da suplementação com simbióticos. Os marcadores inflamatórios foram analisados de duas diferentes maneiras: o grupo todo e considerando a idade como co-variável. Quando comparados os grupos, algumas diferenças foram encontradas em função do tempo para ambos os grupos entre os níveis de IL-10 antes (a) e depois (d) da suplementação (p<0,01) (PLAa=3,9±5,4; PLAd=1,4±1,7; SIMa1,9±3,2; SIMd=1,5±1,3) e IL-6 (p<0,01) (PLAa=3,9±5,4; PLAb=2,4±1,6 e SIMa=3,0±1,9; SIMb=2,5±1,1). A contagem de células sanguíneas revelou maior número de basófilos (contagem relativa) (p=0,01) (PLA=4,22±17,91; SIM=32,05±39,85) e monócitos (contagem absoluta) (p=0,04) (PLA=14,58±8,60; SIM=11,00±7,36) no grupo SIM comparado ao grupo PLA. Conclusão: Considerando tudo o que foi exposto, é possível concluir que a suplementação com simbióticos pareceu ser eficiente para idosos considerados em risco de fragilidade, mas ainda é necessário que mais estudos sejam desenvolvidos incluindo populações maiores, e análises de permeabilidade intestinal.
Background: Ageing is associated to a reduction in the capacity to cope with stressors, which is the main concept of frailty. A number of changes in body composition and function are related to the development of frailty, such as: visceral deposition of fat (highly associated with systemic inflammation) and reduction in the subcutaneous fat. A remarkable change is related to a systemic and low level rise in inflammatory markers in the blood, which has been associated to a broader process called inflammaging. Our work highlights the hypothesis that this low-level inflammation is associated to an imbalance in gut microbiota and changes in intestinal permeability, which in turn justifies the use of synbiotic substances. Aims: to evaluate the effect of a 6-month supplementation of a synbiotic substance on the systemic inflammation in elderly in risk of frailty. Methods: we studied 49 individuals, 65-90 years old, fulfilling from one to two frailty criteria proposed by Fried et al (2001). A double-blind randomized clinical trial was performed, in which the participants were allocated in one of the following groups: SYN (symbiotic treatment) - intake of a synbiotic substance (Frutooligossacarideos 6 g, Lactobacillus paracasei 109 to 108 CFU, Lactobacillus rhamnosus 109 to 108 CFU, Lactobacillus acidophilus 109 to 108 CFU and Bifidobacterium lactis 109 to 108 CFU); or PLA (placebo, control)- maltodextrin in the same doses as the SYN group. Both groups were instructed to consume the substances twice a day, for the period of 6 months. Before and after the supplementation the subjects were evaluated for: plasma inflammatory markers (IL-6, IL-10 and TNF-α); blood cells count; anthropometric measures, gut function (Bristol Scale and Roma criteria). Data were analyzed by repeated measures ANOVA (inflammatory markers), with appropriated pos-hoc test, student\'s t Test (blood cells), as well as odds ratio and risk reduction analysis. The Intention to treat principle was adopted. Significance was considered as p<0.05. Results: The PLA group was composed by 24 subjects with mean age of 76.2±8.4 years, and the PLA group was formed by 24 subjects with mean age 75.6±8.1 years. The gut function was improved in 5.3% of SYN. In turn, PLA group presented 20% of negative symptoms of gut function. The OR of improvement of SYN according to ROMA III was 0.76 (CI= 0.09-6.17), indicating a benefit of the synbiotic supplementation. The inflammatory markers were analyzed in two different ways: the whole group and considering the age as co-variable. When compared the whole groups, some differences were found by the time between both groups for IL-10 at baseline (b) and after supplementation (a) for IL-10 (p<0,01) (PLAb=3,9±5,4; PLAa=1,4±1,7; and SYNb1,9±3,2; SYNa=1,5±1,3) and IL6 (p<0,01) (PLAb=3,9±5,4; PLAa=2,4±1,6 and SYNb=3,0±1,9; SYNa=2,5±1,1). The blood cells count revealed higher numbers of basophiles (p=0,01; PLA=4,22±17,91; SYN=32,05±39,85) and monocytes (p=0,04; PLA=14,58±8,60; SYN=11,00±7,36) on the SIM group compared to the PLA group. Conclusion: Taking altogether, it\'s possible to conclude that the synbiotic supplementation seemed to be efficient for the elderly considered in risk of frailty, but it remains necessary to perform more studies, including larger populations and with analysis of gut permeability.

The objective of this work was to adapt a synbiotic aerated diet dessert, produced with the addition of a probiotic culture of Lactobacillus acidophilus La-5 and prebiotic ingredients (fructooligosaccharides and inulin), from the previously developed sucrose-containing formulation, and to evaluate the effects of its ingestion on adult volunteers with metabolic syndrome (MetS) during a period of 8 weeks of intervention. In addition, to improve the resistance of the probiotic to simulated gastrointestinal conditions, a microencapsulation process was optimized. For the development of the product, the formulations were produced in triplicates, in which probiotic culture survival, instrumental texture and sensory acceptability were evaluated up to 112 days of storage under freezing (-18 °C). Subsequently, a randomized, double-blind, placebo-controlled trial was carried out in which the product developed was administered to forty-five volunteers with MetS assigned into two groups, each receiving 40 g/day of: synbiotic diet mousse (SDM) (n=23) and placebo diet mousse (PDM) without pro- and prebiotics (n=22). Fasting blood samples were collected at the beginning and after 8 weeks of daily consumption of both mousses to determine the anthropometric, biochemical, haematological, inflammatory, and immunological parameters. Afterward, with the goal of improving the survival of L. acidophilus La-5 to in vitro simulated gastrointestinal conditions, the microencapsulation process conditions of the probiotic strain via spray drying were optimized using inulin as the encapsulating agent. The viability of L. acidophilus La-5 incorporated into SDM was above 7.8 log CFU/g and remained stable throughout storage. PDM showed lower acceptability (5.77-6.50) after storage than SDM (6.67-7.03). The texture was the most appreciated attribute and hardness of the SDM increased during storage, but remained stable for PDM. The clinical trial revealed significant reductions of total cholesterol and HDL-cholesterol, as well as of immunoglobulins (A and M), and interleukin-1β in both groups during the intervention period. However, regarding intergroup changes, there were not any significant differences for all parameters evaluated (p>0.05). After the optimization of the microencapsulation process of the probiotic culture (80 mL/min, 82% and 10%, respectively for feed flow, aspiration rate, and inulin concentration), the microencapsulated probiotic strain incorporated in the SDM mousse showed the highest in vitro gastrointestinal survival (p<0.05) in the different stages of the assay, as follows: after the gastric phase: 5.68 log CFU/g (83.3%), the enteral phase I: 5.61 log CFU/g (82.3%), the enteral phase II: 5.56 log CFU/g (81.4%). Therefore, these results suggest that the presence of probiotic and prebiotics in SDM did not provide an additional effect on the health of volunteers with MetS. Additionally, the results confirm the appropriateness of the spray drying process to microencapsulate L. acidophilus La-5 using inulin as coating agent, providing increased resistance to the microencapsulated probiotic strain under in vitro gastrointestinal stress.
O objetivo deste trabalho foi adaptar uma sobremesa aerada simbiótica diet do tipo musse, processada com a adição de uma cultura probiótica de Lactobacillus acidophilus La-5 e de ingredientes prebióticos (fruto-oligossacarídeos e inulina), a partir da formulação contendo sacarose desenvolvida anteriormente, e avaliar os efeitos de sua ingestão em voluntários adultos com síndrome metabólica (MetS) durante um período de 8 semanas de intervenção. Adicionalmente, para melhorar a resistência do probiótico frente às condições gastrintestinais simuladas, otimizou-se um processo de microencapsulação da cepa probiótica. Para o desenvolvimento do produto, as formulações foram produzidas em triplicata, em que se avaliou a sobrevivência da cultura probiótica, a textura instrumental e a aceitabilidade sensorial até 112 dias de armazenamento sob congelamento (-18 oC). Em seguida, foi realizado um estudo randomizado, duplo-cego e controlado por placebo, no qual o produto desenvolvido foi administrado a quarenta e cinco indivíduos com MetS divididos em dois grupos, cada um recebendo 40 g/dia de: mousse simbiótica diet (SDM) (n=23) e musse placebo diet (PDM) sem componentes pro- e prebióticos (n=22). As amostras sanguíneas foram coletadas em jejum no início e após 8 semanas de consumo diário de ambas as musses para a determinação dos parâmetros antropométricos, bioquímicos, hematológicos, inflamatórios e imunológicos. Posteriormente, com o intuito de melhorar a sobrevivência do L. acidophilus La-5 em condições gastrointestinais simuladas in vitro, as condições de processo de microencapsulação da cepa probiótica via spray drying foram otimizadas, utilizando inulina como agente encapsulante. A viabilidade de L. acidophilus La-5 incorporados na SDM foi superior a 7,8 log UFC/g e se manteve estável ao longo do armazenamento. A PDM mostrou menor aceitabilidade (5.77-6.50) após o armazenamento do que a SDM (6.67-7.03). A textura foi o atributo mais apreciado, sendo que a dureza da SDM apresentou elevação, enquanto a da PDM manteve-se estável. O ensaio clínico revelou reduções significativas de colesterol total, colesterol-HDL, imunoglobulinas (A e M) e interleucina1β em ambos os grupos durante o período de intervenção. Entretanto, no que se refere às mudanças intergrupos, não se observou diferenças significativas para todos os parâmetros avaliados (p>0,05). Após a otimização do processo de microencapsulação da cultura probiótica (80 mL/min, 82% e 10%, respectivamente para o fluxo de alimentação, taxa de aspiração e concentração de inulina), a cepa probiótica microencapsulada incorporada a amostra SDM apresentou a maior sobrevivência gastrointestinal in vitro (p<0,05) nas diferentes etapas do ensaio, a saber: após a fase gástrica: 5,68 log UFC/g (83,3%); fase entérica I: 5,61 log UFC/g (82,3%); fase entérica II: 5,56 log UFC/g (81,4%). Portanto, esses resultados sugerem que a presença de probiótico e prebiótico na SDM não apresentou efeitos adicionais na saúde dos voluntários com MetS. Adicionalmente, os resultados confirmaram a adequação do processo de spray drying para a microencapsulação de L. acidophilus La-5 utilizando inulina como agente de revestimento, proporcionando uma maior resistência da cepa probiótica microencapsulada às condições gastrintestinais simuladas in vitro.

The aim of this project was to evaluate the effects of probiotics, prebiotics and synbiotics on the gut ecosystem, immune function and growth parameters of broiler. The first study screened naturally occurring Campylobacter levels in four local sites and revealed the NTU broiler research unit and the NTU animal unit laying hens were Campylobacter free, but a small holding with laying hens was positive and the commercial broiler farm was negative until thinning, after which it was positive. The second study investigated possible delivery routes of a novel strain of Lactobacillus johnsonii (FI9785) into broiler chicken gut and concluded feed was the optimum method for delivery. A third study compared the effect L. Johnsonii FI9785 supplied via feed to control and showed no significant difference in the CFU of caecal Campylobacter, no significant (p≤0.05) effects on growth performance and serum uric acid concentration over 4 weeks. However, mucin layer thickness in the jejunum was significantly (P≤0.05) increased. Concentration of IgA in the serum blood of probiotic treated birds was also increased but IgM and IgG were not significantly altered. Study 4 involved isolation and in vitro screening of candidate probiotic isolates of lactic acid bacteria and a prebiotic from Jerusalem artichoke plant (JA). All tests confirmed the isolates had the characteristics of lactic acid bacteria and have an inhibition activity toward Campylobacter. All isolates belonged to the genus of Lactobacillus and all retained viability during freezing and drying and the poultry gastrointestinal environment, indicating all were potential probiotic agents. Assessment of JA inulin levels indicated the plant to be a potentially good prebiotic source with these isolates. Study 5 investigated in vivo effects of the Lactobacillus isolates (probiotic), JA powder (prebiotic), synbiotic (mix of pre and probiotic). Caecal content were negative for Campylobacter throughout but at day 7, abundance of Firmicutes phyla were higher (p≤0.05) than control for all of supplements treatments and abundance of Faecalibacterium genus numerically increased in all treatments but significantly (p≤0.05) only in 5% prebiotic and probiotic supplemented diets. At day 42, abundance of genus of Erysipelotrichaceae decreased in all treatments. Assessment of growth performance showed JA had no effects but probiotic and synbiotic supplementation caused a degradation in the body weight and increased feed intake. Supplements downregulated the cytokine expression IFNγ,IL-10 and IL-6 in the ileum tissue but showed no effect in the bursa tissue.

The gut microbiota and its metabolites – namely short chain fatty acids (SCFAs) – interact with the digestive, immune, and nervous systems. Microbiota with disrupted composition are highly associated with obesity, gastrointestinal symptoms, and chronic inflammation. Levels of SCFAs in the feces can represent dynamics of the microbiota, and they represent one mechanism by which the microbiota interacts with its host. This study aimed to further our understanding of associations between microbiota bacterial diversity and SCFAs, immune markers, BMI, and GI symptoms and to identify bacteria that are differentially abundant in different BMI groups and with synbiotic supplementation. Data (SCFAs, immunoglobulins, body mass index, fecal fiber, fecal protein, measures of GI symptoms, and 16s RNA sequences, n=11) was extracted from a randomized control trial investigating the effects of synbiotic supplementation in non-celiac gluten-sensitive participants. QIIME2 was used to process 16s RNA data, analyze quantitative, qualitative, phylogenetic quantitative, and phylogenetic qualitative measures of alpha and beta diversity and to perform an analysis of composition of microbiomes (ANCOM) for identification of differential abundances. Multiple metrics of alpha diversity were found to significantly correlate with IgG4, IgM, IL-2, acetate, propionate, isobutyrate, valerate, isovalerate, caproate, heartburn, urgent need to defecate, and feelings of incomplete evacuation. Multiple metrics of beta diversity were significantly different between normal and overweight, normal and obese, and overweight and obese BMI classification groups. Beta diversity was also found to significantly correlate with IgG1, IgG3, IgG4, IgA, IL-6, IL-8, fecal fiber, propionate, butyrate, heartburn, acid regurgitation, nausea and vomiting, bloating, abdominal distension, increased gas, and eructation. The synbiotic intervention did not significantly alter alpha or beta diversity. An ANCOM identified bacterial taxa differentially abundant with BMI shifts and synbiotic supplementation, though these taxa were not those included in the synbiotic. Findings demonstrate alpha and beta diversity associations with various SCFAs, GI symptoms, immune markers, and BMI, and the results of the placebo-controlled intervention suggest careful consideration of placebo contents moving forward. This research supports plans to apply analysis to larger sample sizes to elucidate changes microbial profiles that are associated with clinically relevant biomarkers and symptoms.

The microbial populations in the gastrointestinal tracts of poultry play an important role in normal digestive processes and in maintaining animal health. The purpose of this study was to evaluate the effects of probiotics, prebiotics and synbiotics on the growth parameters, gut ecosystem, histology and immune function. In this study, four experiments one in vitro and three in vivo were conducted using specific pathogen free (SPF) and Hubbard broiler chickens. The first experiment was designed to determine the influence of inulin as an effective prebiotic on lactic acid bacteria (LAB) strains, and to screen LAB for selection as a source of chicken probiotic. Eight strains of LAB were isolated from chicken caeca and three strains from the Plymouth University culture collection were screened for potential probiotic properties for growth in inulin from Jerusalem artichoke (Helianthus tuberosus) and commercial inulin (Frutafit® HD, Netherlands). Lactobacillus animalis JCM 8692 strain isolated from chicken caeca showed the highest auto-aggregation and co-aggregation ability, resistance to acidity and bile salts, strong suppression of pathogens and ability to adhere to epithelial cells compared with other isolated strains. The second experiment was conducted to investigate the influence of commercial inulin and Jerusalem artichoke tubers as prebiotic supplementation on the diversity of the caecal microflora, jejunum histology and immune organ of SPF chickens. This investigation has found that inulin which was extracted from JA had a similar result when compared with commercial inulin and could be a suitable candidate for an inulin source in broiler diets. The third experiment was conducted to investigate the influence of Bactocell® (PRO1) and Lb. animalis (PRO2) as probiotic supplements on broiler chickens. EPEF was significantly increased in probiotic1 and probiotic2 compared with control (311.03, 309.87 and 260.06) respectively. Both types of probiotics supported the growth of chicks healthy and could be a suitable candidate as a source of probiotic in broiler diet. The fourth experiment was conducted to investigate the influence of dietary supplementation of a probiotic (Lb. animalis), a prebiotic JA tuber and a combination of both (Synbiotic) in broiler chickens. Growth performance was improved in all additive supplementation compared with the control group. EPEF was increased in probiotic, prebiotic and synbiotic compared with control (290.8±11.8, 300.9±3.86, 322.1±7.09 and 262.3±5.94) respectively. Beneficial bacteria in the guts of chicks fed probiotic, prebiotic and synbiotic was increased compared with chicks fed control diet. The diversity of microbial population in the gastrointestinal tract of chickens improved due to additives. The intestinal villus lengths and microvilli density was improved in all additives supplementation in comparison with control. Overall, it was concluded that probiotic, prebiotic and synbiotics can positively affect production performance and can improve the gut health.

O presente trabalho teve como objetivo verificar a viabilidade da cepa probiótica Bifidobacterium animalis subsp. lactis Bb-12 incorporado em margarina, suplementada com inulina, concentrado protéico de soro (WPC) e concentrado de caseína (CMP), bem como avaliar as características do produto e a resistência do probiótico às condições simuladas do trato gastrintestinal humano. Foram produzidos 7 diferentes tipos de margarinas de mesa (60% de lipídios: 60 % de óleo de palma + 40% de óleo de canola), empregando-se um modelo de mistura, onde inulina, WPC e CMP foram as variáveis estudadas. Uma formulação controle foi produzida (M8), sem adição desses ingredientes. A utilização da mistura do óleo de palma com óleo de canola favoreceu nutricionalmente as formulações, fornecendo produtos contendo ácidos graxos essenciais em sua composição e ausência de ácidos graxos trans. As formulações M1 a M7, exceto a formulação M2 após o 21º dia de armazenamento, apresentaram populações satisfatórias de Bb-12 para um alimento probiótico, com populações acima de 6 log UFC/g durante 35 dias de armazenamento. Margarinas suplementadas com inulina apresentaram populações satisfatórias durante todo o armazenamento, atingindo populações de 8,01 log UFC/g ao 35º dia (M1). Além disso, M3 e M6, revelaram populações de Bb-12 de 6,87 log UFC/g e 7,27 log UFC/g (dia 35), respectivamente. Por outro lado, M8 não foi caracterizada como margarina probiótica, uma vez que apresentou populações abaixo de 6 log UFC/g, já ao 1º dia de armazenamento. Embora WPC seja utilizado em pesquisas para aumentar a viabilidade de probióticos em alimentos, a suplementação de margarina com WPC sem inulina ou CMP não resultou em populações satisfatórias de Bb-12, apresentando decréscimo de 7,82 (dia 1) para 4,64 log UFC/g (M2, dia 35) (p<0,05). Durante todo o ensaio de resistência in vitro, Bb-12 apresentou sobrevivência significativamente superior (p<0,05) em M1 e revelou populações acima de 6 log UFC/g após 6h de ensaio mesmo ao 28º dia. As populações observadas para M2 diminuíram drasticamente durante o ensaio in vitro (5 log UFC/g após 2h no dia 7). Para as outras formulações, as populações de Bb-12 diminuíram 2 log UFC/g após 2h de ensaio in vitro. Entretanto, M1, M2 e M5 (dias 14 e 28) revelaram aumento significativo nas populações de Bb-12 (p<0,05) entre a fase gástrica (2h) e a segunda fase entérica (6h). As margarinas suplementadas com inulina, principalmente M1, revelaram decréscimo significativo no pH durante todo o armazenamento (p<0,05). Entretanto, isto não afetou a qualidade sensorial dos produtos, uma vez que não foram detectadas diferenças significativas entre as formulações após 7 e 14 dias de armazenamento (p>0,05). A suplementação de margarina com inulina e CMP garantiu populações apropriadas de Bb-12 durante o armazenamento estudado pelo menos até o 28º dia. Além disso, contribuiu para sua sobrevivência durante o ensaio de resistência in vitro. Os resultados revelaram que a margarina apresenta-se como uma matriz alimentar adequada para administração de Bb-12, principalmente quando a inulina foi adicionada.
This study aimed to determine the viability of probiotic Bifidobacterium animalis subsp. lactis Bb-12 incorporated in margarine, with inulin, whey protein concentrate (WPC) and caseinomacropeptide (CMP) supplementation. In addition, the in vitro resistance of Bb-12 incorporated in margarine and related properties were evaluated. Seven margarine-making trials (60% of fat: 60% of palm oil +40% canola oil) were produced, using a mixture model, where inulin, WPC and CMP were the variables studied. Also, a control formulation without these ingredients was manufactured. The use of blending palm oil with canola oil improved the margarine formulations nutritionally, providing products containing essential fatty acids in its composition and absence of trans fatty acids. The formulations M1 to M7, except M2 after 21 days of storage, revealed satisfactory Bb-12 populations for a probiotic food, with counts above 6 log CFU/g during 35 days of storage at 5±1ºC. Margarines supplemented with inulin presented suitable Bb-12 populations throughout the whole storage period, reaching up to 8 log CFU/g by the end of storage (M1). Also, M3 and M6, revealed Bb-12 populations of 6.87 log CFU/g and of 7.27 log CFU/g (day 35), respectively. In contrast, M8 was not characterized as probiotic margarine, since it showed Bb-12 populations below 6 log CFU/g on day 1. Even though whey protein is largely employed in probiotic foods, margarine supplementation with WPC without inulin or CMP did not lead to Bb-12 satisfactory populations, decreasing from 7.82 (day 1) to 4.64 log CFU/g (M2, day 35) (p<0.05). During the whole in vitro assays, Bb-12 survived significantly better (p<0.05) in M1 and revealed populations above 6 log CFU/g after 6h even after 28 days. M2 populations decreased drastically during the in vitro assays for all storage period tested (reduction of 5 log CFU/g after 2h of in vitro assays on day 7 and populations of 2.8 log CFU/g after 6h). For the other formulations, Bb-12 populations decreased 2 log CFU/g after 2h of the in vitro assays. However, for M1, M2 and M5 (on day 14 and 28) the populations of Bb-12 increased significantly (p<0.05) between the gastric phase (2h) and the enteric phase (6h). Formulations containing inulin, mainly M1, showed a significant decrease in pH values during the whole storage period (p<0.05). However, this ingredient did not affect the sensory quality of products, since no significant differences between formulations after 7 and 14 days of storage were observed (p>0.05). The supplementation of margarine with inulin and CMP guaranteed appropriate Bb-12 populations during storage for at least 28 days, and also contributed for its survival throughout the in vitro assays. Therefore, margarine might be considered an appropriate food matrix for Bb-12 survival, mainly when inulin is also added.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-03T19:19:19Z No. of bitstreams: 1 2015 - Marina Jorge de Lemos.pdf: 6290720 bytes, checksum: 466915088ab49af16fcd11435ccc2913 (MD5)
Made available in DSpace on 2017-05-03T19:19:19Z (GMT). No. of bitstreams: 1 2015 - Marina Jorge de Lemos.pdf: 6290720 bytes, checksum: 466915088ab49af16fcd11435ccc2913 (MD5) Previous issue date: 2015-12-08
CAPES
An experiment was conducted in order to evaluate the intestinal morphology, performance, egg quality and analysis of the financial feasibility of including various additives in feed during the whole production period of Japanese quails. 360 quails were distributed in a completely randomized design in a time split plot scheme (9 to 23 weeks of age and 24 to 39 weeks old) with five treatments and eight repetitions of nine birds each. The used treatments were: control, antibiotic, prebiotic, probiotic and synbiotic. The intestinal morphology was evaluated by the height and width and width/height ratio of the villus, crypt depth and villus/crypt ratio; performance, by the feed intake, egg production, egg weight average, feed conversion (kg / kg and kg / dozen), egg mass and viability; egg quality, by the Haugh unit, yolk index, percentage of egg components, shell thickness measured by the micrometer and the scanning electron microscope (SEM); financial analysis was computed by the internal rate of return, net present value and cost benefit. The inclusion of additives in the feed increased the height and width of the villi, decreased crypt depth and increased villous / crypt ratio compared to the control. The inclusion of the additives reduced the feed intake in the two periods of production of the quails in comparison with the control. Throughout the production period, consumption was lower after inclusion of antibiotic and synbiotic in the feed. Egg production, egg weight average, egg mass and feed conversion (kg / kg and kg / dozen) were improved in the quails fed with additives in the two periods of production. The supply of additives within 24 to 39 weeks of age increased the Haugh unit of the eggs compared to the control. The inclusion of additives in the feed increased the yolk and shell percentage, and improved shell and membrane thickness measured by the micrometer compared to the control in both periods of production. The shell thickness (SEM) and the shell palisade layer were improved after inclusion of antibiotic and synbiotic in the feed within 24 to 39 weeks of age. The financial analysis showed best values in the treatments that used antibiotics and synbiotic. The inclusion of antibiotics, probiotic, prebiotic and synbiotic in the feed improves performance and egg quality throughout the production phase of Japanese quails, and these improvements increase effects when the birds receive the additives in the first productive period. The inclusion of antibiotics and synbiotic is more effective than the inclusion of the other additives in reducing food intake throughout the production period and improving shell thickness during the second production period. Intestinal integrity of Japanese quails improved after the inclusion of the tested additives. The financial analysis proved to be more viable to include antibiotic or synbiotic in the feed of Japanese quails in comparison to the other studied additives.
Foi conduzido um experimento com o objetivo de avaliar a morfologia intestinal, o desempenho, a qualidade dos ovos e an?lise da viabilidade financeira da inclus?o de diferentes aditivos na ra??o durante todo o per?odo produtivo de codornas japonesas. Foram utilizadas 360 codornas distribu?das em um delineamento inteiramente casualizado em esquema de parcela subdividida no tempo (de 9 at? 23 semanas de idade e de 24 at? 39 semanas de idade) com cinco tratamentos e oito repeti??es de nove aves cada. Os tratamentos utilizados foram: controle, antibi?tico, prebi?tico, probi?tico e simbi?tico. A morfologia intestinal foi avaliada atrav?s da altura e largura e propor??o largura/altura das vilosidades, profundidade de cripta e rela??o vilo: cripta; o desempenho pelo consumo de ra??o, produ??o de ovos, peso m?dio dos ovos, convers?o alimentar (kg/kg e kg/d?zia), massa dos ovos e viabilidade; a qualidade de ovos pela unidade Haugh, ?ndice de gema, percentagem dos componentes do ovo, espessura de casca medida pelo micr?metro e pelo microsc?pio eletr?nico de varredura (MEV); a an?lise financeira foi computada pela taxa interna de retorno, valor presente l?quido e benef?cio custo. A inclus?o dos aditivos na ra??o aumentou a altura e largura das vilosidades, diminuiu a profundidade de cripta e aumentou a rela??o vilo:cripta em compara??o com o controle. A inclus?o dos aditivos reduziu o consumo de ra??o nos dois per?odos de produ??o das codornas em compara??o com o controle. Durante todo o per?odo produtivo, o consumo foi menor ap?s inclus?o de antibi?tico e simbi?tico na ra??o. A produ??o de ovos, o peso m?dio dos ovos, a massa de ovos e a convers?o alimentar (kg/kg e kg/d?zia) foram melhores nas codornas alimentadas com os aditivos nos dois per?odos de produ??o. O fornecimento dos aditivos no per?odo de 24 at? 39 semanas de idade aumentou a unidade Haugh dos ovos em rela??o ao controle. A inclus?o dos aditivos na ra??o aumentou a porcentagem de gema e de casca, e melhorou a espessura da casca avaliada pelo micr?metro e das membranas da casca em compara??o com o controle nos dois per?odos de produ??o. A espessura da casca (MEV) e da camada pali?ada da casca foi melhor ap?s inclus?o de antibi?tico e simbi?tico na ra??o no per?odo de 24 at? 39 semanas de idade. A an?lise financeira mostrou os melhores valores nos tratamentos que utilizaram antibi?tico e simbi?tico. A inclus?o de antibi?tico, probi?tico, prebi?tico e simbi?tico na ra??o melhora o desempenho e a qualidade dos ovos durante toda a fase produtiva de codornas japonesas, sendo esse efeito potencializado quando as aves recebem os aditivos no primeiro per?odo produtivo. A inclus?o de antibi?tico e simbi?tico ? mais eficaz que a inclus?o dos outros aditivos na redu??o do consumo de ra??o em todo per?odo produtivo e em melhorar a espessura da casca durante o segundo per?odo produtivo. A integridade intestinal das codornas japonesas melhora ap?s a inclus?o dos aditivos testados. A an?lise financeira demonstrou ser mais vi?vel a inclus?o de antibi?tico ou de simbi?tico na ra??o das codornas japonesas em compara??o com os outros aditivos estudados.

O objetivo do presente trabalho foi avaliar o efeito da adição de inulina (I-X1), concentrado proteico de soro de leite (WC-X2) e/ou isolado proteico de soro de leite (WI-X3) em sorvete simbiótico de açaí (Euterpe oleracea Mart.) sobre a viabilidade do probiótico L. rhamnosus GG e a sua sobrevivência frente às condições encontradas no trato gastrintestinal simuladas in vitro, bem como sobre as características tecnológicas e a aceitabilidade sensorial dos produtos resultantes, ao longo de seu armazenamento a -18°C por até 112 dias. Adicionalmente, os sorvetes de açaí foram caracterizados quanto à sua composição centesimal, ácidos graxos insaturados e potencial antioxidante. Por último, objetivou-se a otimização do sorvete simbiótico de açaí a partir dos parâmetros de dureza instrumental e fração de derretimento. Para este fim, foi empregado um delineamento experimental para misturas simples (centroid simplex) incluindo um ponto axial, utilizando diferentes proporções dos fatores X1, X2 e X3. No total, 9 formulações foram avaliadas por um período de até 112 dias de armazenamento congelado. As populações de L. rhamnosus GG permaneceram estáveis e entre 8 e 9 log ufc/g para todas as formulações e ao longo de todo o período de armazenamento. Entretanto, houve redução em, pelo menos, 5 ciclos logaritmos da população do probiótico, após 6 horas de simulação gastrintestinal em todos os períodos estudados, com uma recuperação de L. rhamnosus GG próxima a 4 log ufc/g. Apenas ao 7º dia de armazenamento, a resistência in vitro do probiótico nas formulações contendo inulina, WC e WI superou em até 0,93 log ufc/g a formulação controle. A composição de ácidos graxos revelou que o ácido oleico esteve em maior quantidade nas formulações (teor médio 46,47%), seguido do ácido palmitoleico (teor médio 42,12%). Os sorvetes apresentaram atividade antioxidante na faixa de 302,03 a 505,41 µmol TE/g. A maior atividade antioxidante foi observada para a formulação controle, seguida das formulações WC (4% de WC) e WI (4% de WI). As formulações WC (4% WC) e WI (4% WI) apresentaram os maiores valores de dureza, revelando que os fatores WC e WI exerceram forte influência na dureza dos produtos (p<0,05). As menores taxas de derretimento foram observadas nas formulações WI (4% de WI) e WC-WI (2% de WC; 2% de WI). Exceto para a formulação I-WC-WI (p<0,05), a presença de inulina nas demais formulações não alterou de forma significativa a velocidade de derretimento quando comprada à amostra controle. As formulações I-WI (2% de inulina; 2% de WI) e I-WC-WI (1,33% de inulina; 1,33% WC; 1,33 WI), apresentaram os maiores valores de overrun (37,95% e 39,18%, respectivamente). Os escores de aceitabilidade sensorial variaram de 5,83 a 7,63. Apenas as formulações WI (4% WI) e WC-WI (2% WC; 2% WI) aumentaram seus escores médios de aceitabilidade durante os 84 dias de armazenamento (p<0,05). De acordo com os modelos obtidos, WC foi o fator que mais contribuiu tanto para o aumento de dureza como para a velocidade de derretimento das formulações. A partir da otimização simultânea, considerando as respostas avaliadas (textura e fração de derretimento), é desejável que a formulação de sorvete simbiótico açaí seja produzida contendo 1,33% de inulina, 1,33% de WC e 1,33% de WI. Os resultados obtidos sugerem que a utilização dos três ingredientes (inulina, WC e WI) pode ser vantajosa no desenvolvimento de um sorvete probiótico de açaí, uma vez que a presença desses ingredientes contribuiu com o valor nutricional das formulações e também melhorou as características tecnológicas de overrun, derretimento e textura.
The aimed of the present study was to evaluate the effect of inulin (I-X1), whey protein concentrate (WC-X2) and whey protein isolate (WI-X3) addition on the viability and in vitro survival of L. rhamnosus GG through in vitro simulated gastrointestinal tract (GIT), technological properties and sensory acceptance of symbiotic açai ice cream. Additionally, açai ice cream were characterized for their chemical composition, unsaturated fatty acids and antioxidant activity. Finally, we aimed to optimized the symbiotic açai ice cream using parameters of hardness and melting rate. For this purpose, an experimental design was used for simple mixture (simplex centroid) including an axial point, using different proportions of X1, X2 and X3, nine formulations of synbiotic açai ice cream were studied during frozen storage up to 112 days. A reduction of at least 5 log cycles of L. rhamnosus GG was observed in all formulations after 6 hours of the in vitro assays for all storage period evaluated. However, L. rhamnosus GG recovery was close to 4 log cfu/g for all formulations studied in evaluated periods. Nevertheless, on the 7th day of storage, the probiotic in vitro resistance in formulations supplemented with inulin, WC, and WI exceeded up to 0.93 log cfu/g the control formulation. The fatty acids composition showed that oleic acid was the highest fatty acid in synbiotic açai ice cream (mean content 46.47%), followed by palmitoleic acid (mean content 42.12%). The antioxidant activity of ice creams ranged from 302.03 to 505.41 µmol TE/g. Formulation C showed the highest antioxidant activity, followed by the formulations WC (WC 4%) and WI (WI 4%). Formulations WC (4% of WC) and WI (4% of WI) showed the highest hardness values, revealing that WC and WI exerted a strong influence on the product firmness. The lowest melting rate values were observed in formulation WI (4% of WI) and WC-WI (2% of WC, 2% of WI). However, the presence of inulin and WC showed strong influence on the melting rate of the ice creams, resulting in higher melting down of these products. I-WI and I-WC-WI formulations presented the highest overrun (37.95 and 39.18%, respectively). The sensory acceptability scores ranged from 5.83 to 7.63. WI and WC-WI formulations increased their mean acceptability scores significantly during 84 days of storage, whereas no significant differences were observed in acceptability throughout storage for the other formulations. According to the obtained models, WC was the factor that most contributed to increase both hardness and melting rate. From the simultaneous optimization, considering the responses evaluated (hardness and melting rate), it is desirable a symbiotic acai ice cream containing 1.33% of inulin, 1.33% of WC and 1.33% of WI. The results suggest that the use of the three ingredients (inulin, WC and WI) may be advantageous in the development of a synbiotic açai ice cream, since the presence of these ingredients contributed to the nutritional value of the formulations and also improved the technological characteristics of overrun, melting rate and texture.

BACKGROUND: Tackling severe acute malnutrition (SAM) is a global public health priority. This thesis explores two major influences on treatment outcomes: -Treatment efficacy -Patient-related risk factors OBJECTIVES: 1. To explore whether a pre/probiotic mixture (Synbiotic2000 Forte™) improves treatment outcomes (nutritional and clinical) in children affected by SAM. 2. To describe long term outcomes from SAM and identify key mortality risk factors. METHODS: All 1024 malnourished children admitted to a therapeutic feeding centre in Malawi from July 2006 to March 2007 were eligible for: The PRONUT study (Pre and PRObiotics in the treatment of severe acute malNUTrition): 795 were recruited into a randomised, double-blind, placebo-controlled trial. They received Readyto- Use Therapeutic Food either with or without Synbiotic2000 Forte™. Primary outcome was nutritional cure (weight-for-height >80% of NCHS median). The FUSAM study (Long term Follow-Up after Severe Acute Malnutrition): all children known to be still alive were followed up ≥1 year post discharge. RESULTS: In PRONUT, nutritional cure was similar in both groups: 54%(215/399) for Synbiotic-enhanced RUTF and 51%(203/396) for controls (p=0.40). Main secondary outcomes were also similar (p>0.05). Overall mortality from SAM was 41%(427/1024). Mortality was highest during initial inpatient treatment: 23%(238/1024). In FUSAM, 8%(84/1024) more died within 90 days of admission and 10%(105/1024) during long term follow-up. Cox regression identified HIV, low weight-forheight, low mid-upper arm circumference and low weight-for-age as major risk factors for death (p<0.001). CONCLUSIONS: In this high-mortality setting, Synbiotic2000 ForteTM, did not improve clinical or nutritional outcomes from SAM. A more promising strategy to improve outcomes might be to tackle the major risk factors for SAM mortality: HIV and severity of malnutrition disease. It is likely that earlier treatment would be beneficial. This is a focus of current strategies for both HIV and malnutrition. Rollout of such programmes should be supported and their impact on SAM evaluated.

Негоспітальні пневмонії (НП) займають провідне місце в структурі загальної захворюваності серед дитячого населення та залишаються в ряді 10 найважливіших причин смертності в економічно розвинених країнах. Метою дослідження було вивчення впливу синбіотичного препарату на клінічний перебіг негоспітальної пневмонії у дітей дошкільного віку.

Hintergrund: Gestillte Kinder haben im Vergleich zu nicht gestillten Kindern eine geringere Inzidenz von gastrointestinalen Infektionen und atopischen Erkrankungen. Man geht davon aus, dass der gesundheitsfördernde Effekt der Muttermilch teilweise über die intestinale Mikrobiota vermittelt wird. Diese ist in Stillkindern durch eine geringe Diversität und einen hohen Anteil an Bifidobakterien charakterisiert. Neueste Ansätze in der Weiterentwicklung industriell hergestellter Säuglingsnahrung zielen darauf ab, eine intestinale Mikrobiota zu fördern, die der von gestillten Kindern ähnelt. Die Supplementation von Säuglingsnahrung mit Probiotika (lebende Mikroorganismen) oder Präbiotika (unverdauliche Kohlenhydrate, die als Energiesubstrat für probiotische Bakterien dienen) könnte die bifidogene und antipathogene, aber auch immunmodulierende Wirkung der Muttermilch nachahmen. Aufgrund unterschiedlicher Interaktionen mit der Darmmikrobiota und dem Immunsystem fokussiert man mit der gleichzeitigen Gabe von Pro- und Präbiotika (Synbiotika) eine synergistische Wirkung an. Zielstellung und Studiendesign: In einer randomisiert-kontrollierten, klinischen Studie wurde untersucht, ob sich in den ersten drei Lebensmonaten von gesunden und termingerecht geborenen Kindern mit einer Synbiotikum-haltigen Säuglingsnahrung eine intestinale Mikrobiota etabliert, die der von gestillten Kindern gleicht. Das Synbiotikum setzte sich aus Bifidobacterium animalis ssp. lactis CNCM I 3446 (ältere Bezeichnung B. lactis BB-12) und Kuhmilcholigosacchariden zusammen. Die Studie umfasste zwei Gruppen von Kindern, die eine Säuglingsnahrung mit (SYN-Gruppe, n=21) oder ohne Supplement (KON-Gruppe, n=18) erhielten. Gestillte Kinder dienten als Referenz (REF-Gruppe, n=23). Um die Diversität der Bifidobakterien auf Speziesebene umfassend zu charakterisieren, wurden quantitative Real-Time PCR (qPCR)-Verfahren, basierend auf dem single-copy groEL als phylogenetisches Zielgen, zur spezifischen Quantifizierung von zwölf Bifidobakterienspezies in humanen Fäzes entwickelt und validiert. Ergebnisse: Die supplementierte Säuglingsnahrung war gut verträglich und unterstützte eine gesunde Entwicklung; vergleichbare anthropometrische Daten von SYN- und REF-Gruppe. Das Synbiotikum stimulierte selektiv das Wachstum von Laktobazillen und Bifidobakterien. Die Zellzahl für Laktobazillen der SYN-Gruppe war zur REF-Gruppe äquivalent (9,07±0,32 versus 9,90±0,27 log10 Zellen/g Fäzes TM [MW±SEM]; p<0,0019; Äquivalenzdifferenz von 1 log10 Zellen/g Fäzes TM) und höher als in der KON-Gruppe (8,27±0,31 log10 Zellen/g Fäzes TM [MW±SEM]). Die Zellzahl für Bifidobakterien war in der SYN-Gruppe am höchsten (11,54±0,05 versus 11,00±0,17 [REF-Gruppe] und 10,54±0,24 [KON-Gruppe] log10 Zellen/g Fäzes TM [MW±SEM]). In der SYN-Gruppe wurde die höchste Anzahl an Bifidobakterienspezies erfasst (167 mit [128 ohne] B. animalis in 56 Fäzesproben versus 98 und 93 in jeweils 51 Fäzesproben der REF- und KON-Gruppe). Neben Kinder-typischen Spezies wie B. bifidum und B. breve wurden auch Spezies, die für Erwachsene charakteristisch sind (B. adolescentis), häufiger in der SYN-Gruppe als in den Vergleichsgruppen nachgewiesen. Der pH-Wert in Fäzes von Kindern aus der SYN-Gruppe war niedriger als der aus der KON-Gruppe (6,07±0,20 versus 6,45±0,17 [MW±SEM]) und näher an dem von gestillten Kindern mit 5,29±0,12 (MW±SEM). Schlussfolgerung: Die Supplementation einer Säuglingsnahrung mit dem Synbiotikum aus CNCM I-3446 und Kuhmilcholigosacchariden führte zu einer Angleichung in der Zusammensetzung der intestinalen Mikrobiota und des fäkalen pH-Wertes an gestillte Kinder. Die in dieser Arbeit entwickelten groEL-basierten qPCR-Verfahren erlaubten eine spezifische und genaue Analyse der Bifidobakterienpopulation unter dem Einfluss eines Synbiotikums.
Background: Compared to formula-fed infants, breast-fed infants have a reduced incidence of gastrointestinal infections and atopic diseases. The health-promoting effect of breast milk is assumed to be partly mediated by the intestinal microbiota, which is characterized by a low diversity and a high proportion of bifidobacteria. Recent approaches in further development of infant formulae aim at promoting an intestinal microbiota similar to that of breast-fed infants. The supplementation of infant formula with probiotics (live microorganisms) or prebiotics (non-digestible carbohydrates, which serves as energy substrates for probiotic bacteria) could mimic the bifidogenic and antipathogenic, but also immunomodulating effect of breast milk. Due to various interactions with the gut microbiota and the immune system, the simultaneous administration of pro- and prebiotics (synbiotics) is focussed to have a synergistic effect. Objective and study design: In a randomized-controlled, clinical trial healthy full-term infants receiving an infant formula with synbiotic for the first three months of life were studied, whether an intestinal microbiota is induced, which is equivalent to that of breast-fed infants. The synbiotic consisted of Bifidobacterium animalis ssp. lactis CNCM I 3446 (previously known as B. lactis BB-12) and cow milk oligosaccharides. The study comprised two groups of infants receiving a starter formula with (SYN-group, n=21) or without supplement (KON-group, n=18). Breast-fed infants served as a reference (REF-group, n=23). In order to comprehensively characterize the bifidobacteria diversity at species level, quantitative real-time PCR (qPCR) assays based on the single-copy groEL as phylogenetic marker for the specific quantification of twelve bifidobacteria species in human feces were established and validated. Results: The supplemented formula was well tolerated and supported a healthy development; comparable anthropometric data of SYN- and REF-group. The synbiotic selectively stimulated the growth of lactobacilli and bifidobacteria. Lactobacilli levels were equivalent in SYN- and REF-group (9.07±0.32 versus 9.90±0.27 log10 cells/g feces DM [Mean±SEM]; p<0.0019; equivalence margin of 1 log10 cells/g feces DM) and higher than the KON-group (8.27±0.31 log10 cells/g feces DM [Mean±SEM]). The highest levels of bifidobacteria were observed in the SYN-group (11.54±0.05 versus 11.00±0.17 [REF-group] and 10.54±0.24 [KON-group] log10 cells/g feces DM [Mean±SEM]). The highest number of bifidobacteria species were obtained in the SYN-group (167 with [128 without] B. animalis in 56 fecal samples versus 98 and 93 in each of 51 fecal samples of the REF- and KON-group). Beside species, typically found in infants such as B. bifidum und B. breve, also species, which are characteristic for adults (B. adolescentis), were detected more often in the SYN-group than in the other study groups. Fecal pH was lower in the SYN- than in the KON-group 6.07±0.20 versus 6.45±0.17 [Mean±SEM]) and closer to that of breast-fed infants (5.29±0.12 [Mean±SEM]). Conclusion: In infants fed a starter formula supplemented with a synbiotic (CNCM I-3446 and cow milk oligosaccharides), composition of intestinal microbiota and fecal pH were closer to that of breast-fed infants. The groEL-based qPCR-assays, developed in this study, allowed a specific and accurate analysis of the bifidobacterial population in response to the synbiotic intake.

Les probiotiques sont des microorganismes vivants qui, lorsqu’ils sont ingérés en quantité adéquate, peuvent exercer des propriétés bénéfiques sur la santé de l’hôte. Les souches de Bacillus utilisées en tant que probiotiques ne sont pas colonisatrices du tractus intestinal, mais sont des résidents transitoires du microbiote. Ce travail fait l’investigation in vitro de l’association, qualifiée de synbiotique, entre une souche probiotique de Bacillus subtilis et une source carbonée prébiotique, composé alimentaire peu digéré par les enzymes intestinales mais utilisable par les bactéries dans l’intestin. L’étude de cette association met en évidence la capacité de la souche à utiliser les isomaltooligosaccharides (IMOS) prébiotiques comme unique substrat carboné. L’effet positif de ce substrat sur la tolérance à la bile de la souche a été démontré in vitro. Les résultats d’une analyse protéomique faisant l’étude des enzymes clés impliquées dans le métabolisme des IMOS, ainsi que d’autres biomarqueurs d’intérêt probiotique, sont en cours d’exploitation. Ce travail préliminaire d’investigation de l’association synbiotique entre les IMOS prébiotiques et la souche probiotique B. subtilis, aboutira à des essais in vivo. Les effets bénéfiques des probiotiques du genre Bacillus, notamment au niveau de la modulation du système immunitaire, résultent de l’interaction entre les molécules de la surface bactérienne et les cellules de l’intestin. Les mécanismes moléculaires à l’origine de l’immunomodulation sont mal connus, alors que leur compréhension est nécessaire à l’optimisation de l’utilisation du probiotique. Un deuxième volet de ce travail concerne la comparaison des structures d’entités moléculaires de surface de trois Bacilli probiotiques, les acides lipotéichoïques (LTAs), et leurs activités immunologiques respectives. Une étude structurale des LTAs par des méthodes biochimiques et par RMN a permis de mettre en évidence la diversité structurale au sein du même genre Bacillus. Le rôle clé de la D-alanine dans l’activité biologique de ces antigènes bactériens a été démontré
Probiotics are live microorganisms, which when administered in adequate amounts confer a health benefit on the host. Bacillus probiotic strains are not able to colonize the gut, and are considered as transient residents of the microbiota. Prebiotic are non-digestible food ingredients that could stimulate growth of bacteria in the gut. This work investigates the in vitro effect of a prebiotic isomaltooligosaccharide (IMOS) on the growth of a probiotic strain Bacillus subtilis. This study highlights the ability of the strain to use IMOS as unique carbon source. A comparative proteomic analysis investigates the main enzymes implicated in the prebiotic metabolism, and biomarkers possibly involved in probiotic effects. This preliminary work, which studies the synbiotic association between a probiotic and a prebiotic, will lead to in vivo assays. Beneficial effects of probiotic Bacilli, mainly modulation of intestinal immune system, result from interaction between bacterial cell-wall molecules and intestinal cells. The molecular origin of immunomodulatory mechanisms are poorly understood, while understanding is needed to optimize the use of probiotics. A second part of this work consists in comparing the structure of a molecular cell-wall component named lipoteichoic acids (LTA) from three Bacillus probiotic, a molecular cell-wall component of Gram positive bacteria, and their immunological activity. A structural study, using biochemical determinations and NMR spectroscopic analysis, highlights the structural diversity between LTAs from different Bacillus species. The key role of D-alanine substituents in the biological activity of these bacterial antigens has been demonstrated

The aim of this thesis was to study the effect of symbiotic administration on measurable parameters of gut-barrier function and clinical outcome in the following groups of surgical patients;  Elective surgical admissions, Intensive Care patients, Irritable Bowel Syndrome sufferers and Crohn’s Disease patients. Each study was performed as a double blind, randomised and placebo controlled trial.  The symbiotic preparation contained oligofructose and the probiotic bacteria Lactobacillus acidophilus La5, Bifidobacterium lactis Bb-12, Streptococcus thermophilus and Lactobacillus bulgaricus. The preoperative ingestion of synbiotics had no influence on the rate of bacterial translocation, endotoxin exposure or subsequent septic morbidity in elective surgical patients.  Nasogastric colonisation by potentially pathogenic organisms was significantly reduced in the population of critically ill patients after symbiotic therapy, but was not associated with improved clinical outcome.  Physical and psychological symptom scores improved in both the placebo and active symbiotic groups of irritable Bowel Syndrome patients, representing a significant placebo response.  Synbiotic dietary supplementation had no significant effect on relapse rate, markers of disease activity or nutritional status in patients with Crohn’s Disease over a one year period. Synbiotic administration significantly altered nasogastric colonisation by potentially pathogenic bacteria, but had no effect on any other quantifiable measure of gut-barrier function.  The clinical significance of this finding is uncertain, but warrants further investigation.  To date, there is insufficient scientific evidence to recommend the routine use of synbiotics in surgical patients.

Diversos estudos têm mostrado que alimentos adicionados de probióticos e prebióticos, em quantidades suficientes, trazem benefícios à saúde humana. Estes benefícios se devem, fundamentalmente, ao seu efeito na manutenção da microbiota intestinal benéfica. Paralelamente, ocorre aumento do consumo de derivados do açaí em grandes centros urbanos no Brasil e até mesmo no exterior, principalmente a sua polpa e derivados, que se deve, principalmente, ao alto valor calórico da fruta e à presença de pigmentos antioxidantes. O presente trabalho teve como principal objetivo desenvolver um mix de açaí que apresente aspectos nutricionais e sensoriais semelhantes ao produto tradicional, porém com propriedade funcional suplementar, por se tratar de um alimento probiótico, prebiótico ou simbiótico. Quatro tratamentos foram produzidos (três vezes cada um deles), todos contendo mix de açaí congelado: M1 (controle), M2 (com L. acidophilus La-5 + B. animalis subsp. lactis Bb-12), M3 (com inulina) e M4 (com L. acidophilus La-5 + B. animalis subsp. lactis Bb-12 + inulina) e armazenados a -18 °C por até 3 meses. Após 1, 7, 14, 21, 28, 56 e 84 dias de armazenamento, foram determinadas as populações de microrganismos probióticos, o pH e a cor instrumental dos produtos. O teor de umidade foi determinado após a fabricação dos produtos (dia 0). Adicionalmente, após 7, 42 e 84 dias de armazenamento, foi realizada análise sensorial (aceitabilidade). Foi determinada a composição centesimal dos produtos, a partir de amostras mantidas liofilizadas. Todos os resultados foram comparados, através de análise estatística. A diferenciação na composição centesimal e no teor de sólidos entre as formulações decorreu da adição da fibra prebiótica. Com relação à evolução da cor e do pH, não houve variação importante ao longo do tempo, assim como entre os diferentes mixes de açaí estudados. Os produtos obtiveram boa aceitabilidade sensorial por parte dos provadores (médias das notas acima de 7, na maioria das formulações). Houve maior aceitabilidade dos mixes suplementados com o ingrediente prebiótico e o mix simbiótico recebeu notas médias significativamente mais elevadas (p < 0,05) em dois períodos de armazenamento (7 e 84 dias), quando comparado aos demais. O tempo de armazenamento não influenciou a avaliação sensorial significativamente (p > 0,05). Durante o período de armazenamento proposto, os probióticos apresentaram sobrevivência satisfatória (populações superiores a 108 UFC por porção diária), sob o ponto de vista da legislação brasileira vigente, com exceção do B. animalis subsp. lactis no mix não suplementado de inulina. O mix de açaí apresentou-se como boa matriz para veicular os microrganismos probióticos testados e a inulina contribuiu para uma maior aceitabilidade sensorial do produto.
Some studies have been showing that food products supplemented with probiotics and prebiotics, in sufficient amounts, result in benefits to human health. These benefits are related, fundamentally, to their effect on the healthy intestinal microbiota maintenance. Concomitantly, consumption of açai food products, mainly its pulp and derivatives, is increasing in urban centers in Brazil and also abroad. The high caloric value and the presence of antioxidant pigments are responsible for its higher consumption. The main aim of the present study was to develop an açai mix with nutritional and sensory aspects similar to the conventional product, but with a supplementary functional property, by being a probiotic, prebiotic or synbiotic food. Four trials where produced (three times each), all containing frozen açaimix: M1 (control), M2 (supplemented with L. acidophilus La-5 + B. animalis subsp. lactis Bb-12), M3 (supplemented with inulin) and M4 (supplemented with L. acidophilus La-5 + B. animalis subsp. lactis Bb-12 + inulin) and stored at -18 °C for up to 3 months. After 1, 7, 14, 21, 28, 56 and 84 days of storage, the products were analyzed for probiotic microorganisms populations, pH and instrumental color. The moisture content was determined soon after production (on day 0). Additionally, after 7, 42 and 84 days of production, sensory evaluation (acceptability test) was carried out. Finally, compositional analyses proceeded from freeze-dried samples. Results were compared through statistical analysis. Differences in compositional analyses and moisture content between trials were attributed to the presence or absence of inulin. Instrumental color and pH evolution were not influenced by the storage period and also no differences between the different acaimixes studied were observed (p > 0,05). Products were well accepted by the panellists, and mean scores were often above 7. Prebiotic fiber inulin improved overall acceptability of açaimixes and the synbiotic mix obtained higher mean score (p < 0.05) in two storage periods (7 and 84 days), when compared to the others. The storage period did not influence sensory evaluation significantly (p > 0.05). During the storage period studied, probiotics showed a satisfactory survival (viability higher than 108 cfu for the products daily portion), according to the current Brazilian legislation, except for B. animalis subsp. lactis in the mix not supplemented with inulin. The açaimix showed to be a good matrix for the probiotic microorganisms tested and inulin contributed for a higher acceptability of the product.

Immediately post-hatch, broiler chicks are exposed to microbes that begin colonizing the gut, including environmental pathogens. One of the costliest enteric diseases in broiler production is necrotic enteritis (NE), caused by the ubiquitous opportunistic bacteria Clostridium perfringens (CP). With the worldwide reduction in antibiotic growth-promoters (AGPs), there is increased interest in natural alternatives to reduce disease and improve broiler health. The overall objective of the studies described herein was to apply probiotics or synbiotics to birds by in ovo application or orally before they leave the hatchery, then evaluate bird performance and various intestinal responses. Data were analyzed in JMP with LS Means to separate means with significance assigned at P ≤ 0.05 and trends at 0.05 < P ≤ 0.10. The first 21-day (D) study used 480 male Cobb 500 broilers randomly divided into one of four treatments using a 2x2 factorial design: a no-additive control (CTRL), a one-time oral application of synbiotic at the hatchery fed a basal diet (HS), an oral application of water at the hatchery with dietary synbiotics (DS), and a hatchery synbiotic plus dietary synbiotic (HSDS). Performance was measured on day-of-hatch (DOH), D3, D7, D14, and D21. mRNA abundance of various intestinal markers was measured at D7 and D21, including tight junction proteins ZO-1, ZO-2, and CLD-1; nutrient transporters SGLT1 and PepT1; and immune response markers TLR2, TLR4, and IL-10. HS lowered feed intake (FI) and feed conversion ratio (FCR) without lowering body weight (BW) from D14-21. There was greater abundance of PepT1 mRNA (P ≤ 0.1) and IL-10 mRNA (P ≤ 0.05) on D21 in HSDS. Second, a 21-day pilot study with 480 male and female Cobb 500 broilers was conducted to determine the optimum in ovo dosage level of a probiotic or synbiotic (PROB or SYNB) applied at embryonic day 18 (E18) with subsequent NE challenge using seven treatments: in ovo application of sterile water (CTRL), low (PROB-L or SYNB-L: 1x105 CFU), medium (PROB-M or SYNB-M: 1x106 CFU), or high (PROB-H or SYNB-H: 1x107 CFU) probiotic or synbiotic doses dissolved in sterile water. Performance measurements were taken on DOH, D4, D8, D14 and D21. On D8, NE lesion scores were not impacted by treatment. D8 ileal samples were taken for mRNA abundance of TLR4, IL-10, IL-1β, AvBD8, AvBD10, and AvBD13. SYNB-H had higher abundance of AvBD10 mRNA compared to CTRL (P ≤ 0.1), and higher IL-1β mRNA compared to SYNB-L (P ≤ 0.05). PROB-H and SYNB-H had better performance than the low and medium doses, but were not better than the CTRL. The high doses were chosen for subsequent studies. Third, a longer 42-day study using 1,630 Ross 708 male and female broilers was conducted consisting of the following six treatments. A negative control (NC): sterile water in ovo fed basal corn/soybean meal mash diet without NE challenge; antibiotic growth-promoter (AGP+): sterile water in ovo fed basal diet with virginiamycin (0.5 kg/MT) as an AGP with NE challenge; NC+: same as NC plus NE challenge; SI+: synbiotic in ovo fed the basal diet and NE challenged; SD+: sterile water in ovo fed basal diet supplemented with synbiotic (0.5 kg/MT feed) and NE challenged; and SID+: synbiotic in ovo fed basal diet with synbiotic (0.5 kg/MT feed) with NE challenge. Cumulatively, SID+ had lower FI and FCR than NC+, but no change in BW or BWG. The combination treatment (SID+) often had an additive effect compared to SD+ or SI+ alone on mRNA abundance and D7 cecal fatty acid profiles. SD+ and SID+ also had higher D42 lean:fat ratios compared to NC+. Last, a 42-day study was conducted using 1,630 male and female Ross 708 broilers and the in ovo application of probiotics and subsequent NE challenge with five treatments. NC: sterile water in ovo, fed basal corn/soybean meal mash diet without NE challenge; AGP+: sterile water in ovo, fed basal diet with virginiamycin (0.5 kg/MT of feed) as AGP with NE challenge; NC+: NC treatment, with NE challenge; PI+: probiotic in ovo, fed basal diet, with NE challenge; PD+: sterile water in ovo, fed basal diet supplemented with probiotic (1.3 kg/MT of feed), with NE challenge. The use of probiotics in this study had little effect on performance, lean:fat ratios, and cecal fatty acid profiles, but PD+ increased mRNA abundance of D14 TLR2, D14 TNF-α, and D42 LEAP2 in cecal tonsils compared to controls. PI+ increased mRNA abundance of D7 and D42 MUC2, D7 LEAP2, and D42 TNF-α in the ileum. PI+ increased mRNA abundance in the cecal tonsils of D7 TLR2 and D42 TNF-α. These studies yielded interesting results about probiotics and synbiotics during a NE challenge by evaluating performance, intestinal immune responses, and fatty acid profiles in the ceca of broilers. In conclusion, the probiotic in this study did not improve broiler health during a NE challenge, but synbiotic use in ovo and continuation in the feed showed improvement over in ovo or dietary application alone. Synbiotic improved FCR over a challenged control, and altered mRNA abundance in the small intestine.
Doctor of Philosophy
The poultry industry is one of the most popular animal protein sources worldwide. As with any livestock operation, industry goals include optimizing animal health and well-being, maximizing animal productivity, and producing quality products in the most cost effective manner. Improvements in genetics, nutrition, and management have increased productivity and cut costs. One important application was the low-level use of antibiotics in feed. These medications reduced the risk of disease outbreak in flocks, which led to healthier birds and improved growth rates. However, when global concern of antibiotic resistance in human medicine came to light, both the livestock industry and governing bodies implemented voluntary and mandatory reduction or elimination of antibiotics. Previously, these important antibiotics helped to control costly diseases. As they are removed, alternatives to antibiotics will be important in disease control and prevention. A major group of alternatives to antibiotics in poultry includes probiotics, prebiotics, and synbiotics. Probiotic bacteria are considered 'good bacteria' in the gut, and provide various health benefits to the host. Prebiotics are non-living substances that support the growth of healthy bacteria. A synbiotic is the combination of both probiotics and prebiotics in a single application method. The goal of this research project was to give probiotics or synbiotics to broiler chicks and evaluate their potential benefits and effects on bird performance and the immune response. Ideally, applying probiotic bacteria as early as possible might translate into early colonization of the gut with healthy bacteria. This included oral application of synbiotics at the hatchery, or by safely injecting them into part of the egg that is swallowed by the chick embryo before hatch. This egg application, or in ovo application, is a safe, effective, widely-practiced method of vaccinating chicks to jumpstart their defense against disease. By vaccinating them in ovo, they can start to prime the immune system before they even hatch. Applying probiotics in ovo may improve health after early gut colonization with beneficial microbes. Numerous studies on natural alternatives to antibiotics have been conducted, with varying results. Results of this research indicate that in ovo application of probiotics and synbiotics is safe. Birds that received probiotics in the feed often performed similar to those that received none. However, the in ovo use of synbiotics combined with the continued use in the feed after hatch improved efficiency in broilers during an intestinal disease challenge and improved various aspects of gut function. Overall, as antibiotics are phased out, using probiotics and synbiotics may improve poultry health, but continued research will help understand the optimum ways to use them.

Thesis (PhD)--Stellenbosch University, 2014.
ENGLISH ABSTRACT: Background: Synbiotics, probiotics and prebiotics are being added to infant formula. This study was an in-depth evaluation of research on infants fed infant formula containing synbiotics, probiotics or prebiotics and was carried out in two phases. Phase one included two systematic reviews that assessed if synbiotics, probiotics or prebiotics led to improved growth and clinical outcomes in formula fed full term and preterm infants. Phase two included two studies: A systematic review compared the methodological quality and outcomes of industry and non-industry sponsored randomized controlled trials (RCTs) and a descriptive study evaluated how the food industry applies the knowledge and evidence gained from probiotics, prebiotics or synbiotics research in infants. The research questions were: Does the consumption of probiotics, prebiotics or synbiotics supplemented infant formula lead to improved clinical outcomes in infants? Is there an association between source of funding and methodological quality, clinical outcomes and author’s conclusions in trials using probiotics, prebiotics or synbiotics supplemented formula in infants? Does the food industry use the evidence gained through probiotics, prebiotics and synbiotics research trials on infants for the benefit of the general paediatric population? The hypotheses were: Consumption of probiotics, prebiotics or synbiotics by infants leads to improved clinical outcomes; The source of funding in research trials using probiotics, prebiotics or synbiotics supplemented formula in infants is associated with outcomes in favour of the sponsor’s products and authors’ conclusions; Methodological qualities of non-industry sponsored trials are equivalent to industry sponsored trials; Evidence gathered through probiotics, prebiotics and synbiotics research is implemented by the food industry. Methods: Phase one: Both systematic reviews on preterm and full term infants: Cochrane methodology was followed using RCTs which compared preterm or full term formula containing probiotics, prebiotics or synbiotics to conventional infant formula with / without placebo among healthy preterm or full term infants. The mean difference (MD) and corresponding 95% confidence intervals (CI) were reported for continuous outcomes, risk ratio (RR) and corresponding 95% CI for dichotomous outcomes. Phase two: In the systematic review, Cochrane methodology was used to assess the risk of bias of included RCTs. Association between source of funding and risk of bias, clinical outcomes and conclusions were assessed. In the descriptive study, all listed companies that manufacture infant food products with added synbiotics, probiotics or prebiotics for infants were identified and invited to participate. A letter of invitation was sent and if they expressed willingness to take part in the study, a questionnaire with a written consent form was sent. Descriptive statistics and associations between categorical variables were to be tested using a Chi-square test. Results: Phase one: Review on preterm infants: 8 studies were included. Probiotics increased stool frequency with no effect on other clinical outcomes. Prebiotics increased stool frequency and bifidobacteria counts only. Review on full term infants: 25 studies were included. Synbiotics improved stool frequency but had no effect on other clinical outcomes. Probiotics did not have an effect on any clinical outcome. Prebiotics increased weight gain and stool frequency with no effect on other outcomes. Phase two: Systematic review: 67 studies were included, majority were funded by food industry. There was no significant association between the source of funding and four domains (sequence generation, allocation concealment, blinding, selective reporting), majority of reported clinical outcomes or authors’ conclusions. Source of funding was significantly associated with two domains (incomplete outcome data, free of other bias), antibiotic use and conclusions on weight gain. Descriptive study: 25 companies were identified and invited to participate. No company agreed to participate in the survey for different reasons. Conclusions Phase one: Review on preterm infants: There is not enough evidence to state that supplementation with probiotics or prebiotics results in improved growth and clinical outcomes in exclusively formula fed preterm infants. Review on full term infants: There is not enough evidence to state that supplementation of term infant formula with synbiotics, probiotics or prebiotics does result in improved growth or clinical outcomes in term infants. There is no data available to establish if synbiotics are superior to probiotics or prebiotics. Phase two: Systematic review: In RCTs on infants fed infant formula containing probiotics, prebiotics or synbiotics, the source of funding does not influence majority of outcomes in favour of the sponsors’ products. More non-industry funded research is needed to further assess the impact of funding on reported clinical outcomes and authors’ conclusions. Descriptive study: Due to companies refusing to participate in this study, no conclusion could be drawn on how the food industry applies evidence gained through probiotics, prebiotics or synbiotics research on infants. More transparency is needed from the infant formula manufactures on how they apply the evidence gained from probiotic, prebiotic or synbiotic research on infants.
AFRIKAANSE OPSOMMING: Agtergrond Synbiotika, probiotika en prebiotika word gereeld by baba formule gevoeg. Hierdie studie was 'n in-diepte evaluering van navorsing oor babas gevoed met formule melk wat synbiotika, probiotika of prebiotika bevat en is uitgevoer in twee fases. Fase een het twee sistematiese oorsigte ingesluit wat die rol van synbiotika, probiotika en prebiotika op verbeterde groei en kliniese uitkomste van formule gevoede volterm babas en vroeg gebore babas evalueer het. Fase twee het bestaan uit twee studies: 'n sistematiese oorsig wat die metodologiese kwaliteit en uitkomste van die bedryf en nie-bedryf geborgde ewekansige gekontroleerde proewe (RCTs) evalueer het, asook 'n beskrywende studie wat die kennis en toepassing van bewyse oor die effektiewiteit van probiotika, prebiotika of synbiotika in die voedsel industrie bestudeer het. Die hipotese stellings was: verbruik van probiotika, prebiotika of synbiotika by babas lei tot verbeterde kliniese uitkomste; die bron van befondsing vir synbiotics, probiotika of prebiotika navorsing beïnvloed uitkomste ten gunste van die borg se produkte; bewyse ingesamel deur middel van probiotika, prebiotika en synbiotika navorsing word geïmplementeer deur die voedselindustrie. Metodes Fase een: Beide sistematiese oorsigte op volterm en premature babas: Cochrane metodes is gevolg deur ewekansige, gekontroleerde studies wat vol termyn of premature formule met probiotika, prebiotika of synbiotika met konvensionele baba formule met / sonder plasebo onder gesonde volterm of premature babas bestudeer. Die gemiddelde verskil (MD) en die ooreenstemmende 95% vertrouensintervalle is gebruik vir deurlopende uitkomste, risiko verhouding (RR) en die ooreenstemmende 95% CI vir tweeledige uitkomste. Fase twee: In die sistematiese oorsig is Cochrane metodiek gebruik om die risiko van vooroordeel van ingesluite ewekansige, gekontroleerde studies te evalueer. Assosiasie tussen bron van befondsing en die risiko van vooroordeel, asook kliniese uitkomste en gevolgtrekkings was beoordeel. In die beskrywende studie, is alle genoteerde maatskappye wat babavoeding produkte vervaardig met bygevoegde synbiotika, probiotika of prebiotika vir babas geïdentifiseer en uitgenooi om deel te neem. 'n Uitnodigingsbrief is vir die relevante maatskappye gestuur om hul bereidwilligheid om deel te neem te bevestig. Indien hulle wel bereid was om deel te neem was 'n vraelys met 'n skriftelike toestemming vorm gestuur. Beskrywende statistiek en assosiasies tussen kategoriese veranderlikes was getoets met behulp van 'n Chi-kwadraat toets. Resultate Fase een: Oorsig oor premature babas: 8 studies was ingesluit. Probiotika verhoog stoelgang frekwensie met geen effek op ander kliniese uitkomste. Prebiotika verhoog ook stoelgang frekwensie en slegs bifidobakteriële tellings. Oorsig oor die vol termyn babas: 25 studies was ingesluit. Synbiotika verbeter stoelgang frekwensie, maar het geen effek op ander kliniese uitkomste gehad nie. Probiotika het nie 'n effek op enige kliniese uitkoms gehad nie. Prebiotika verhoog gewigstoename en stoelgang frekwensie met geen effek op ander uitkomste. Fase twee: Sistematiese oorsig: 67 studies was ingesluit, en die meerderheid was befonds deur die voedsel bedryf. Daar was geen beduidende assosiasie tussen die bron van befondsing en vier gebiede (toekenningsvolgorde, toekenningsverberging, studie verblinding, selektiewe verslaggewing), en die meerderheid van gerapporteerde kliniese uitkomste of skrywers se gevolgtrekkings. Die bron van befondsing was beduidend verbind met twee gebiede (onvolledige uitslag data, vry van ander vooroordeel), antibiotika gebruik en gevolgtrekkings op gewigstoename. Beskrywende studie: 25 maatskappye is geïdentifiseer en genooi om deel te neem. Geen maatskappy het ingestem om deel te neem aan die studie om verskillende redes. Gevolgtrekkings Fase een: Oorsig oor premature babas: Daar is nie genoeg bewyse dat die aanvulling met probiotika of prebiotika resultate in verbeterde groei en kliniese uitkomste in uitsluitlik formule gevoede premature babas tot gevolg het nie. Oorsig oor die volle termyn babas: Daar is nie genoeg bewyse om te sê dat die aanvulling van term baba formule met synbiotika, probiotika of prebiotika lei tot verbeterde groei of kliniese uitkomste in termyn babas. Daar is geen inligting beskikbaar om te stel of synbiotika beter is as probiotika of prebiotika nie. Fase twee: Sistematiese oorsig: In studies op babas gevoed met formule melk wat probiotika, prebiotika of synbiotika bevat het, het die bron van befondsing nie meerderheid van die uitkomste in die guns van die borge se produkte beïnvloed nie. Meer nie-industrie befondsde navorsing is nodig om verder die impak van befondsing op kliniese uitkomste en skrywers se gevolgtrekkings te evalueer. Beskrywende studie: Aangesien al die maatskappy deelname geweier het, kon geen gevolgtrekking gemaak word of die voedsel bedryf bewyse oor die gebruik van probiotika, prebiotika of synbiotika toepas nie. Meer deursigtigheid is nodig van die formule vervaardigers oor hoe hulle die bewyse oor die gebruik van probiotika, prebiotika of synbiotika toepas.

Celem badań przedstawionych w niniejszej dysertacji było określenie wpływu substancji prebiotycznych oraz synbiotycznych podanych in ovo (do komory powietrznej) w 12. dobie rozwoju zarodkowego, na poziom ekspresji genów kurcząt brojlerów
The purpose of the study presented in this dissertation was to determine the effects of prebiotic and synbiotic delivered in ovo on the 12th day of embryonic development on the gene expression level of the broiler chickens
Streszcz. ang

Objetivos: O presente trabalho é secundário a um estudo clínico randomizado, que teve por objetivos investigar os efeitos de uma substância simbiótica sobre a permeabilidade intestinal, a inflamação sistêmica, a cognição e sintomas depressivos em idosos. Métodos: quarenta e nove participantes, 65-90 anos de idade, ambos os gêneros, foram randomizados em dois grupos: grupo S (uso de simbióticos); e grupo P (uso de placebo). As seguintes avaliações foram realizadas no início e no final do experimento: Escala de Depressão Geriátrica (GDS-15); Mini Exame do Estado Mental (MEEM); % de gordura corporal (%gordura); citocinas pró-inflamatórias e regulatórias no soro (IL-6, TNF-α e IL-10); e marcadores de permeabilidade intestinal no soro (DAO, IFABP e LPS). Resultados: A interação entre tempo e grupo mostrou maior redução do LPS no grupo P e maior aumento do IL-10 no grupo S. Considerando somente o fator tempo, ambos os grupos reduziram a %gordura, TNF-α e DAO. O IL-6, o GDS-15 e MEEM aumentaram em ambos os grupos. O valor final de GDS-15 foi explicado negativamente por DAO, IL-10, TNF-α, %gordura, ser mulher e estar alocado no grupo P. As variáveis que explicaram positivamente o valor final do GDS-15 foram IL-6, IFABP, e LPS. As variáveis positivamente associadas aos valores finais do MEEM foram IL-10, DAO, ser mulher e estar alocado no grupo P; foram encontradas associações negativas entre o MEEM e IL-6, TNF-α, %gordura, IFABP e LPS. Foi discutida a possibilidade de alguma interferência dos medicamentos utilizados pelos idosos, nas variáveis de interesse do estudo. Conclusões: de acordo com nossas condições experimentais, foram encontrados efeitos sutis da suplementação de simbióticos sobre sintomas depressivos e cognição em idosos da comunidade, aparentemente saudáveis; entretanto, os efeitos sobre a cognição parecem ter sido mais otimistas. Estudos futuros, com indivíduos com diagnóstico de depressão ou de comprometimento cognitivo, poderão clarear questões levantadas no presente estudo.
Objectives: This secondary study from a randomized clinical trial aimed to investigate the effect of a synbiotic substance on intestinal permeability, systemic inflammation, cognition and depressive symptoms in elderly. Methods: Forty-nine participants, 65-90 years old, both genders, were randomly assigned into two groups: S-group (synbiotic); and P-group (placebo). Evaluations performed at the beginning and at the end of the experiment were: Geriatric Depressive Symptoms scale (GDS-15); Mini-Mental Status Examination (MMSE); % of body fat (%fat); pro-inflammatory and regulatory cytokines (serum IL-6, TNF-α and IL-10); gut permeability (serum DAO, IFABP and LPS). Results: The interaction between time and group showed higher reduction of LPS in P-group and higher increase of IL-10 in S-group. Considering only time, both groups had reduced their %Fat, TNF-α and DAO. The IL-6, GDS-15 and MMSE were increased in both groups. The GDS-15 final was negatively explained by DAO, IL-10, TNF-α, %Fat, being woman and being allocated in P-group. The variables that positively explained the GDS-15final were the IL-6, the IFABP, and the LPS. The variables that were positively associated to MMSE final were the IL-10, DAO, being woman, and being allocated in P-group; The negative associations were IL-6, TNF-α, %Fat, IFABP and LPS. We discussed the possibility of some interference of the medicines used by the elderly on our outcomes of interest. Conclusions: our experimental conditions demonstrated weak effects of synbiotic supplementation in the management of depressive symptoms or cognitive decline in apparently healthy elderly living in the community, although the effect on cognition seems to be more optimist. Other comparative studies, with individuals diagnosed as depressive morbidity or cognitive decline, may clarify our hypothesis.

A associação de ingredientes prebióticos com microrganismos probióticos na elaboração de produtos lácteos pode resultar na obtenção de produtos simbióticos, com os efeitos benéficos dos probióticos, aliado ao estímulo seletivo das bifidobactérias endógenas do cólon. Objetivou-se desenvolver um queijo petit-suisse simbiótico, suplementado com os probióticos Lactobacillus acidophilus e Bifidobacterium lactis e os prebióticos inulina, oligofrutose e mel; avaliar a viabilidade dos probióticos e do starter, a concentração de frutanos, a aceitabilidade sensorial e parâmetros físico-químicos durante o seu armazenamento a 4±1°C por até 28 dias; otimizar as misturas envolvendo os prebióticos, garantindo a boa viabilidade probiótica e o potencial efeito prebiótico, associados a características tecnológicas e sensoriais satisfatórias. Para esse fim, foram desenvolvidas formulações de queijo petit-suisse, com Lactobacillus acidophilus e Bifidobacterium lactis, utilizando Streptococcus thermophilus como starter, conforme o delineamento da superfície de resposta para misturas com três fatores: oligofrutose, inulina e mel. Os fatores foram combinados em diferentes proporções, de modo a obter 10% da formulação final, totalizando 7 ensaios e um controle (T1 a T8). As populações dos probióticos estiveram sempre superiores ao recomendado para a promoção dos efeitos benéficos, tendo variado de 7,20 a 7,69 log ufc g-1 (B. lactis) e de 6,08 a 6,99 log ufc g-1 (L. acidophilus). Os valores de pH decresceram e de acidez cresceram ao longo do tempo, devido à atividade dos microrganismos acidificantes presentes, tendo diferido significativamente entre os ensaios (p<0,05). A umidade permaneceu estável e diferiu significativamente entre os ensaios (p<0,05). O ingrediente que mais afetou a textura instrumental foi a inulina, fornecendo produtos mais firmes, mais coesos e com adesividade e gomosidade maiores. Oligofrutose e mel, como ingredientes únicos, produziram queijo petit-suisse com menor firmeza, adesividade e gomosidade, parâmetros estes correlacionados negativamente com o pH nos diferentes produtos estudados, ao contrário do que ocorreu com a coesividade. Os ensaios não diferiram significativamente quanto à aceitabilidade sensorial com painel de consumidores, exceto aos 28 dias de armazenamento (p<0,05). Aqueles contendo oligofrutose (T1) e oligofrutose mais inulina (T4) foram os mais bem aceitos, enquanto o controle (T8) foi o menos aceito. Todos os resultados ficaram entre \"gostei ligeiramente\" e \"gostei moderadamente\". A aceitabilidade foi crescente durante o armazenamento, significativamente (p<0,05) apenas para os ensaios que continham oligofrutose ou inulina sozinhos e sua combinação. Os atributos mais citados pelos consumidores foram o sabor - mais preferido nos ensaios T1 (oligofrutose), T3 (mel) e T5 (oligofrutose mais mel) e menos preferido no T8 (controle), e a textura, sempre menos preferida, em virtude de \"arenosidade\". Todas as combinações dos ensaios estudadas, exceto o T3 (mel), podem ser consideradas prebióticas, partindo-se da recomendação de 4 a 5 g dia-1 de frutanos e com uma porção diária de 100 g de queijo petit-suisse. A Metodologia da Superfície de Resposta aplicada para otimizar a viabilidade dos probióticos, a firmeza instrumental, o teor de frutanos e o custo, resultou na mistura ótima contendo 25% de oligofrutose, 70% de inulina e 5% de mel e desejabilidade global de 99,55%.
Due to the potential synergy between probiotics and prebiotics, dairy products containing a combination of these ingredients are often referred to as synbiotic, providing the beneficial effects of the probiotics combined with the effect of selectively stimulating the growth and/or activity of bifidobacterium in the colon. The purpose of this study was to develop a synbiotic petit-suisse cheese, supplemented with the probiotics Lactobacillus acidophilus and Bifidobacterium lactis and the prebiotics inulin, oligofructose and honey; to monitor the viability of the probiotics and the starter, the concentration of fructans, and also the sensory acceptance and physico-chemical parameters during refrigerated storage (4±1°C) for up to 28 days; to optimize the mixtures involving the prebiotic ingredients, in order to guarantee good probiotic viability and potential prebiotic effects, together with to satisfactory technological and sensory properties. The formulations were developed according to the surface response methodology for mixtures including three factors: oligofructose, inulin and honey. The prebiotics were combined in different proportions (to achieve 10% of total mixture), and 7 trials plus a control trial were obtained (T1 to T8). Probiotic populations were always above the minimum counts suggested for providing health benefits, and varied from 7.20 up to 7.69 log cfu g-1 (B. lactis) and from 6.08 up to 6.99 log cfu g-1 (L. acidophilus). The pH values decreased and the acidity values increased significantly during storage, and were statistically different in the various trials (P<0.05), due to the acidifying effect of the organisms present. The moisture remained stable during storage and also varied statistically in the trials (P<0.05). Instrumental texture profile was greatly influenced by inulin, resulting in firmer, more cohesive and adhesive products, and with higher gumminess. Oligofructose and honey added as sole ingredients produced a less firm, adhesive and gummy cheese. The pH influenced the instrumental firmness, adhesiveness and gumminess negatively, and cohesiveness positively. There was no significant difference in sensory acceptance for the consumer panel between the different trials, except after 28 days of storage (P<0.05). The acceptance was significantly higher for cheeses supplemented with oligofructose (T1) and oligofructose plus inulin (T4), whereas the control (T8) was less accepted. The results always ranged from like slightly to like moderately. Acceptability increased during storage, though significantly (P<0.05) only for cheeses supplemented with oligofructose and/or inulin. Taste was the most preferred attribute for trials T1 (oligofructose), T3 (honey), and T5 (oligofructose plus honey) and less preferred for T8 (control). Texture attribute was always the least preferred, due to the \"dust\" present. All cheese trials, except T3 (honey), can be considered prebiotic, according to the recommended ingestion of 4 to 5 g day -1 of fructans for a 100g daily portion. The response surface methodology, applied to optimize probiotic viability, instrumental firmness, fructans content and cost, resulted in an optimum mixture containing 25% oligofructose, 70% inulin and 5% honey and global desirability of 99.55%.

Submitted by Nelci Antunes de Moura (nelcimoura@gmail.com) on 2016-05-20T00:29:22Z No. of bitstreams: 1 tese Nelci final.pdf: 3449766 bytes, checksum: 82850e9746cb84286bf3b3204344f1e9 (MD5)
Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-05-23T19:39:07Z (GMT) No. of bitstreams: 1 moura_na_dr_bot.pdf: 3449766 bytes, checksum: 82850e9746cb84286bf3b3204344f1e9 (MD5)
Made available in DSpace on 2016-05-23T19:39:07Z (GMT). No. of bitstreams: 1 moura_na_dr_bot.pdf: 3449766 bytes, checksum: 82850e9746cb84286bf3b3204344f1e9 (MD5) Previous issue date: 2015-12-18
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
O ferro heme presente na carne vermelha está associado ao aumento da incidência do câncer colorretal (CCR). O heme pode catalisar a formação de compostos nitrosos e a peroxidação lipídica no lúmen intestinal. No entanto, os efeitos pró-carcinogênicos do heme podem ser inibidos por alguns compostos como os sais de cálcio, clorofila entre outros. Sabe-se que o indol-3-carbinol (I3C), presente nas plantas da família das Brassicas e os simbióticos são compostos promissores na prevenção do câncer de cólon, atuando em via de proliferação, apoptose e modulação da microbiota intestinal. Dessa forma, o objetivo desse estudo foi o de avaliar os efeitos da ingestão de simbiótico (prebiótico inulina associado ao probiótico Bifidobacterium lactis bb-12) e de I3C, isolados ou em associação sobre o processo de carcinogênese de cólon induzido pela 1,2-dimetilhidrazina (DMH) em ratos Wistar alimentados ou não com dieta suplementada com heme. Os animais foram alocados em 9 grupos, os grupos 1 a 8 (n=12) receberam quatro doses de DMH (40 mg/Kg) nas duas semanas iniciais do experimento. Os grupos 1 e 9 (n=12 e 5) receberam ração basal até o final do experimento e os grupos 2 a 8 receberam ração basal suplementada com heme, heme+I3C, heme+simbiótico, heme+I3C+simbiótico, I3C, simbiótico e I3C+simbiótico, respectivamente. A eutanásia ocorreu ao final da 25ª semana. Neste momento foi realizada a coleta do cólon com os respectivos tumores e amostras de fezes do ceco. Em seguida, procedeu-se a medida dos tumores e coleta de amostras para biologia molecular. Após a fixação em formalina tamponada e a retirada dos tumores, realizou-se a contagem de focos de criptas aberrantes (FCA) pela coloração de azul de metileno. Realizou-se a análise histológica dos tumores e a análise da expressão de 95 genes relacionados a via da carcinogênese colônica, pela técnica Taqman Low Density Array, e a expressão proteica da E-caderina, TGFB1 (Transforming growth factor beta 1) e RAF1 (Serine/threonine-protein kinase) por Western Blotting. Foram analisados os índices de proliferação celular e apoptose pelo PCNA (Proliferating cell nuclear antigen) e caspase 3-clivada, respectivamente, tanto nos cólons como em tumores, e a expressão de β-catenina e E-caderina nos tumores, por imunoistoquímica. Células da linhagem Caco-2 foram incubadas com água fecal extraída das fezes do ceco e submetidas a testes de citotoxicidade e genotoxidade pelos testes do MTT (mitochondrial tetrazolium test) e Cometa, respectivamente. Os dados foram comparados utilizando-se o software Sigma Stat 3.5 e Expression Suíte para expressão gênica. Foi observado aumento significativo no número de criptas aberrantes (CA) no grupo que recebeu heme (G2) quando comparado ao grupo que recebeu apenas ração basal (G1). Redução significativa no número de CA foi observada no grupo que recebeu heme+I3C (G3) e heme+simbiótico (G4) quando comparado ao grupo que recebeu heme (G2). O número de FCA totais com ≥ 9 criptas aberrantes foi significativamente menor no grupo que recebeu heme+simbiótico (G4) quando comparado ao grupo que recebeu heme (G2). Entretanto, aumento significativo no número de tumores com mais de 60 mm3 foi observado no grupo suplementado com heme+I3C+simbiótico (G5), quando comparado ao grupo que recebeu heme (G2). Além disso, foi observado aumento significativo na incidência de tumores invasivos no grupo que recebeu heme+I3C+simbiótico (G5) quando comparado ao grupo que recebeu heme (G2). Os tumores do grupo suplementado com heme+I3C+simbiótico (G5) apresentaram baixa expressão dos genes Cdh1, Tgfb1, Appl1 e alta expressão do Raf1, já os tumores do grupo suplementado com heme +I3C (G3) apresentaram baixa expressão do Cdh1. A água fecal do grupo que recebeu heme (G2) apresentou significativamente maior citotoxicidade e genotoxicidade quando comparado ao grupo que recebeu ração basal (G1). Com relação aos tratamentos, a água fecal do grupo que recebeu heme+I3C (G3) e heme e simbiótico (G4) apresentaram água fecal significativamente com menor potencial genotóxico quando comparada ao grupo que recebeu heme (G2). No entanto, o grupo que recebeu heme+I3C+simbiótico (G5) apresentou aumento significativo na genotoxicidade da água fecal. Dessa forma, concluímos que o heme associado a uma dieta com níveis normais de cálcio não é um potente indutor de FCA, mas aumenta a citotoxicidade e genotoxicidade da água fecal. No entanto, tanto o I3C como o simbiótico reduzem os efeitos citotóxicos/genotóxicos da ingestão de heme. Contudo, a associação do heme+I3C+simbiótico apresentou efeito promotor da carcinogênese de cólon.
Colorectal cancer (CRC) is the third most common type of cancer worldwide. Hemin iron, which is found in red meat, catalyzes the formation of carcinogenic N-nitroso compounds and lipid peroxidation end-products in the colon lumen. The procarcinogenic effect of hemin is known to be inhibited by molecules, such as calcium, chlorophyll and others. However, the preventive effect of indole 3-carbinol and synbiotics on colon carcinogenesis remains uninvestigated. The aim of this study was to assess the modifying effects of a synbiotic (inulin+ Bifidobacterium lactis) and/or I3C against dimethylhidrazine (DMH)-induced colon carcinogenesis in hemin-fed male Wistar rats. Nine groups of animals were evaluated. Groups 1–8 received a total of four s.c. DMH injections (40 mg/kg b.w.) over 2 weeks, whereas group 9 was given EDTA solution (vehicle). Two weeks after DMH-initiation, G1 and G9 were fed a basal diet while groups G2, G3, G4, G5, G6, G7 and G8 received a basal diet containing hemin, hemin+I3C, hemin+synbiotic, hemin+I3C+synbiotic, I3C, synbiotic and I3C+synbiotic, respectively, during 23 weeks. At 25 week, all animals were killed and their colons were removed. Cecal contents were collected to determine fecal water cytotoxicity and genotoxicity (DNA damage) in Caco-2 cells. Colon tumors were measured and samples were collected and stored at -800C. The colons were fixed flat in 10% buffered formalin for 24 h and stained with 1.0% methylene blue for classical ACF analysis and quantification. Tumor incidence and multiplicity were assessed after histopathological analysis. Gene and protein expression were determined in tumor samples alone. The total number of aberrant crypts (AC) was significantly higher (p= 0.03) in the hemin group (G2) than in the group fed basal diet (G1). AC number in both hemin+I3C (G3) and hemin+synbiotic (G4) groups was also significantly lower than in the group fed hemin (G2). Tumor volume was higher in the hemin+I3C+ synbiotic (G5) group and invasive adenocarcinoma was more frequent in the hemin+I3C+synbiotic group (G5) than in the group fed hemin (G2). Colon tumor expression analysis showed that in comparison with the group fed hemin (G2), Cdh1, Tgfb1 and Appl1 were downregulated while Raf1 was upregulated in the group hemin+I3C+synbiotic (G5), and Cdh1 was down-regulated in the group hemin+I3C (G3). Fecal water cytotoxicity in the hemin group (G2) was higher than in groups fed basal diet (G1) and hemin+I3C (G3). Fecal water genotoxicity was also significantly higher in the group fed hemin alone (G2) than in the basal diet group (G1), as well as, in groups fed hemin+I3C (G3) and hemin+synbiotics (G4). However, when compared to hemin alone (G2), fecal water from group hemin+I3C+ synbiotics (G5) presented the highest DNA damage levels. Our results suggest that although hemin in a regular-calcium diet was not a powerful ACF promoter, it increased fecal water citotoxicity and genotoxicity. On the other hand, hemin associated with either I3C or synbiotics prevented ACF promotion. Nonetheless, a synergistic interaction among hemin, I3C and synbiotic did promote DMH-induced tumorigenesis.
FAPESP: 2011/23699-4

Orientador: Luís Fernando Barbisan
Resumo: O ferro heme presente na carne vermelha está associado ao aumento da incidência do câncer colorretal (CCR). O heme pode catalisar a formação de compostos nitrosos e a peroxidação lipídica no lúmen intestinal. No entanto, os efeitos pró-carcinogênicos do heme podem ser inibidos por alguns compostos como os sais de cálcio, clorofila entre outros. Sabe-se que o indol-3-carbinol (I3C), presente nas plantas da família das Brassicas e os simbióticos são compostos promissores na prevenção do câncer de cólon, atuando em via de proliferação, apoptose e modulação da microbiota intestinal. Dessa forma, o objetivo desse estudo foi o de avaliar os efeitos da ingestão de simbiótico (prebiótico inulina associado ao probiótico Bifidobacterium lactis bb-12) e de I3C, isolados ou em associação sobre o processo de carcinogênese de cólon induzido pela 1,2-dimetilhidrazina (DMH) em ratos Wistar alimentados ou não com dieta suplementada com heme. Os animais foram alocados em 9 grupos, os grupos 1 a 8 (n=12) receberam quatro doses de DMH (40 mg/Kg) nas duas semanas iniciais do experimento. Os grupos 1 e 9 (n=12 e 5) receberam ração basal até o final do experimento e os grupos 2 a 8 receberam ração basal suplementada com heme, heme+I3C, heme+simbiótico, heme+I3C+simbiótico, I3C, simbiótico e I3C+simbiótico, respectivamente. A eutanásia ocorreu ao final da 25ª semana. Neste momento foi realizada a coleta do cólon com os respectivos tumores e amostras de fezes do ceco. Em seguida, procedeu-se a... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Colorectal cancer (CRC) is the third most common type of cancer worldwide. Hemin iron, which is found in red meat, catalyzes the formation of carcinogenic N-nitroso compounds and lipid peroxidation end-products in the colon lumen. The procarcinogenic effect of hemin is known to be inhibited by molecules, such as calcium, chlorophyll and others. However, the preventive effect of indole 3-carbinol and synbiotics on colon carcinogenesis remains uninvestigated. The aim of this study was to assess the modifying effects of a synbiotic (inulin+ Bifidobacterium lactis) and/or I3C against dimethylhidrazine (DMH)-induced colon carcinogenesis in hemin-fed male Wistar rats. Nine groups of animals were evaluated. Groups 1–8 received a total of four s.c. DMH injections (40 mg/kg b.w.) over 2 weeks, whereas group 9 was given EDTA solution (vehicle). Two weeks after DMH-initiation, G1 and G9 were fed a basal diet while groups G2, G3, G4, G5, G6, G7 and G8 received a basal diet containing hemin, hemin+I3C, hemin+synbiotic, hemin+I3C+synbiotic, I3C, synbiotic and I3C+synbiotic, respectively, during 23 weeks. At 25 week, all animals were killed and their colons were removed. Cecal contents were collected to determine fecal water cytotoxicity and genotoxicity (DNA damage) in Caco-2 cells. Colon tumors were measured and samples were collected and stored at -800C. The colons were fixed flat in 10% buffered formalin for 24 h and stained with 1.0% methylene blue for classical ACF analysis and qua... (Complete abstract click electronic access below)
Doutor

O aumento populacional e a elevação no consumo de alimentos tem aumentado a necessidade de intensificação da aquicultura objetivando maiores produtividades para atendimento à elevação na demanda. Consequentemente, aumentam as chances de ocorrência de surtos epizoóticos dentro dos sistemas de produção, devido ao maior estresse imposto aos animais densidade elevada, manejo constante, transporte, baixa qualidade da água, etc. Alterações no estado fisiológico dos animais fazem com que seu sistema imunológico não consiga combater agentes inócuos em situações normais, mas que se tornam patogênicos nos casos de imunodepressão, levando a grandes perdas econômicas, seja com mão de obra, mortandade, medicamentos, etc. Assim, este estudo avaliou o efeito da administração individual ou conjunta de -glucano e Bacillus subtilis na dieta de juvenis de pacu, Piaractus mesopotamicus. Em um primeiro ensaio, quantidades crescentes de -glucano (60; 120; 180; 240 e 300 mg kg-1 de ração) e de B. subtilis (109 UFC g-1 de B. subtilis; 0,025%, 0,050%, 0,075%, 0,100%, 0,125%) foram adicionados a uma dieta basal extrudada (32% de proteína bruta PB; 3200 kcal kg-1 de energia digestível ED) e administrados a juvenis de pacu (8,91 ± 0,30 g) até a saciedade aparente por 75 dias, em um delineamento inteiramente aleatorizado. Ao final do período experimental, os animais foram anestesiados, pesados, submetidos à coleta de sangue e tecidos, para determinação do ganho de peso, conversão alimentar, taxa de crescimento específico, proteínas totais, albumina e globulina séricas, índice albumina:globulina (A:G), atividade da lisozima sérica, explosão respiratória de leucócitos e morfometria intestinais. Em um segundo ensaio conduzido nas mesmas condições ambientais, juvenis de pacu (2,0 ± 0,1 g; 15 peixes/aquário) foram alimentados por 59 dias com rações suplementadas pelos aditivos em questão, mais um grupo controle: -glucano de levedura 240 mg kg-1 de ração; B. subtilis comercial (109 UFC B. subtilis g-1) 0,100% de inclusão; tratamento simbiótico 240 mg de -glucano kg-1 + 0,100% de B. subtilis. Ao final do período experimental os peixes foram amostrados para determinação dos mesmos parâmetros anteriormente descritos. A inclusão dos níveis de -glucano e de Bacillus subtilis na dieta não influenciou o desempenho (P>0,05) de juvenis de pacu, mas peixes alimentados com a dieta contendo o B. subtilis consumiram mais ração (P<0,05). Não houve alterações nos parâmetros de morfologia intestinal (P>0,05), nem nos valores de explosão respiratória, atividade de lisozima, proteínas totais séricas e globulinas séricas. Quando os dois imunoestimulantes foram adicionados às dietas em combinação, também não foi observado efeito sobre os parâmetros de desempenho, explosão respiratória e atividade de lisozima, mas as proteinas totais séricas e as globulinas séricas foram positivamente influenciadas pelos tratamentos (P<0,05). As alturas de microvilosidades da porção cranial do intestino dos pacus alimentados com o tratamento simbiótico (-glucano + B. subtilis) foram maiores se comparadas ao grupo controle (P<0,05). Apesar de algumas respostas terem indicado efeito positivo sobre alguns parâmetros imunológicos e morfologia intestinal de pacus alimentados por uma combinação de -glucano + B. subtilis, não houve clara evidência que a administração simbiótica é superior à administração individual de cada imunoestimulante.
Stimulation of immune response by dietary supplements has already been proven effective in aquaculture. This work evaluates effects of dietary supplementation with the prebiotic -glucan, the probiotic Bacillus subtilis and their synbiotic combination on growth performance, immune responses, and intestinal morphology of pacu Piaractus mesopotamicus. In a first trial, a basal diet (32% crude protein; 3200 kcal kg-1) was supplemented with either -glucan (60; 120; 180; 240 e 300 mg kg-1) and B. subtilis (109 UFC g-1 of B. subtilis; 0.025%, 0.050%, 0.075%, 0.100%, 0.125%) levels and fed to groups of juvenile pacu (eight fish; 8.91 ± 0.3 g) stocked in 33 glass aquariums (70 L) for 75 days, setting up a totally randomized design trial (n=3). Fish were then weighed and had their blood drawn for determination of leucocytes respiratory burst activity, serum lysozyme activity and serum immunoglobulin concentration; intestinal tissue was sampled from two fish from each unit for micromorphology analysis. The inclusion of -glucan and Bacillus subtilis in the diet did not affect growth (P>0.05) of pacu juvenile, but fish fed diet containing levels of B. subtilis showed higher feed consumption (P<0.05). Intestinal morphology, respiratory burst activity, lysozyme activity, total serum proteins and total globulins were not affected in either inclusion of -glucan and Bacillus subtilis (P>0.05). In a second trial, a basal diet (34% crude protein; 3400 kcal kg-1) was supplemented with either -glucan (240 mg kg-1), B. subtilis (0.1 %) and -glucan + B. subtilis mix and fed to groups of juvenile pacu (15 fish; 2.0 ± 0.1 g) stocked in 16 glass aquariums (70 L) for 59 days, setting up a totally randomized design trial (n=4). Fish fed diets supplemented with -glucan + B. subtilis mix presented better growth rate, but it was not significantly different from other feeding groups. Serum total proteins and serum globulins (serum total proteins serum albumin) were affected by dietary supplements (P < 0.05); fish receiving the synbiotic treatment had higher serum globulin contents, but respiratory burst and lysozyme activity were not significantly affected. Dietary synbiotic mix altered the micromorphology of anterior intestine (P < 0.05), but the same effect was not observed in the mid and posterior intestines (P > 0.05). Dietary -glucan + B. subtilis synbiotic mix affected the intestinal ultrastructure of pacu, improved responses of immune system but did not significantly affect growth rate.

De nombreuses pathologies découlent de déséquilibres du microbiote intestinal. Ainsi, des stratégies visent à les prévenir en restaurant cet écosystème par l'apport de probiotiques, de prébiotiques et de synbiotiques. Des résultats prometteurs nécessitant d'être validés rationnellement, nous nous sommes proposés d'établir un premier crible de sélection de bactéries lactiques et de glucooligosaccharides (GOS) pour des activités anti-inflammatoires et anti-prolifératives in vitro. Les probiotiques ont inhibé des pathogènes tout en se révélant résistants aux conditions du tube digestif, tandis que deux GOS ont conduit à leur croissance sélective. De plus, les probiotiques ont réduit la réponse inflammatoire de cellules intestinales et la prolifération de cellules cancéreuses en association avec un GOS. Nous avons donc retenu des produits potentiellement actifs contre les maladies inflammatoires chroniques de l'intestin et du cancer colorectal, par un crible devant être validé in vivo
Some pathologies are induced by intestinal microbiota disorders. Thus, some strategies aim torestore this ecosystem through probiotics, prebiotics and synbiotics. Promising results need tobe rationally validated, so we aimed to establish the screening first step of lactic acid bacteriaand glucooligosaccharides (GOS) against anti-inflammatory and anti-proliferative activity invitro. Probiotics inhibited pathogens and were resistant to digestive tract conditions whileGOS promoted specifically their growth. Moreover, probiotics reduced inflammatoryresponse of intestinal cells and proliferation of cancer cells when combined with GOS. Thus,we selected compounds potentially efficient against inflammatory bowel diseases andcolorectal cancer, through a screen that need to be validated in vivo

Thesis (MScAgric)--Stellenbosch University, 2008.
ENGLISH ABSTRACT: Different substances were evaluated and compared to an antibiotic, in terms of their effect on nitrogen - and amino acid digestibilities. Two digestibility trials and one performance trial were conducted. Trials one and two apparent nitrogen (AND)- and amino acid (AAD) digestibilities were determined from digesta collected at the terminal ileum (ileal digestibility method). In Trial 3 the substances were evaluated in terms of their potential to improve production performance. Broilers were fed a maizesoya based diet throughout the three trails. In the first trial, garlic and a commercial prebiotic (Bio-Mos®), were tested and compared in terms of AND and AAD, to an antibiotic (doxycyclin, Doxyvete-SOS). A starter and finisher diet were fed as either mash or pellets. The garlic was included at 8g/kg, 13g/kg and 18g/kg to the starter and finisher diets. Bio-Mos® was added at 1g/kg, 2g/kg and 3g/kg to the starter diet, and 0.5g/kg, 1g/kg and 1.5g/kg to the finisher diet. The doxycyclin was added at 0.3 g/kg. None of the treatments had any beneficial effects in terms of AND. Feeding a pellet seem to have some negative effects in terms of AND. In general most of the treatments did not show any improvement in AAD at any determination period (day 21, 28 or 35). At day 21 and day 35, the mash diet supplemented with 18g/kg garlic had a negative effect on AAD, when compared to the negative and positive control. It doesn’t seem that feeding either a mash or a pellet had an influence on the effects exerted by the different treatments. In the second trial the influence of Bio-Mos®, a blend of organic acids, probiotics and electrolytes (Acid-Pak 4-way®) and a medium-chain triglyceride (MCT) were evaluated and compared in terms of AAD and AND, to the effect of an antibiotic, doxycyclin. The starter and finisher diets were fed as a mash. Bio-Mos® was included at 1g/kg, 2g/kg, and 3g/kg in the starter diet, and at 0.5g/kg, 1g/kg, 1.5g/kg in the finisher diet, respectively. Acid-Pak 4-way® was included at 0.4g/kg, 1g/kg and 1.6g/kg for both the starter and finisher diets. Medium-chain triglycerides (MCT) were allocated at 3g/kg, 3.6g/kg, 4.2g/kg for the starter diet, and 2.1g/kg, 2.7g/kg and 3.4g/kg for the finisher diet. An antibiotic, doxycyclin, was included at 0.3 g/kg. With AND, no treatment had any significant effect for the entire experimental period. At day 21, the treatment supplemented with MCT (3.4g/kg) had the most significant beneficial effect on AAD, when compared to the negative- and positive controls, as it increased AAD for the majority of the amino acids. The treatment with Acid-Pak 4-way® (1g/kg) had the most significant negative effect on AAD when compared to the positive control. At day 28, the treatments with Bio-Mos® (0.5g/kg and 1.5g/kg) and Acid-Pak 4-way® (0.4g/kg) had the most significant beneficial effect on AAD when compared to the positive control. It increased AAD for more than half of the 17 amino acids evaluated. The treatment supplemented with MCT (2.7g/kg) has shown the most significant negative effect on AAD, when compared to the positive control. In the third trial the effect of Bio-Mos®, Acid-Pak 4-way® and MCT on production performance was evaluated, and compared to the effects of the presence or absence of doxycyclin. Body weight (BW), body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR) were measured. The starter and finisher diets were fed as a mash. Bio-Mos®, MCT and Acid-Pak 4-way® were included at 3.0g/kg, 4.2g/kg and 1.6g/kg, respectively in the starter and finisher diets. Birds were weighed (per pen) on arrival and on days 7, 14, 21, 28, 35. Feed intake (FI) per pen was measured at days 7, 14, 21, 28 and 35, and mortality was recorded daily. In terms of BWG, Acid-Pak 4-way® had a higher BWG, when compared to the negative control, Bio-Mos® and MCT. It can be concluded that Bio-Mos®, Acid-Pak 4-way®, as well as MCT can be a possible alternatives to antibiotic supplementation. These three treatments did not necessary prove to be more effective than antibiotics, but are definitely competitive alternatives.
AFRIKAANSE OPSOMMING: Verskillende behandelings is geëvalueer en vergelyk met ‘n antimikrobiese produk, in terme van hul uitwerking op stikstof - en aminosuur verteerbaarhede. Twee verteringstudies en produksieprestasie studie is uitgevoer. In die eerste twee studies is die skynbare stikstof (AND)- en aminosuur (AAD) verteringskoöeffisiënte bepaal deur gebruik te maak van digesta wat by die terminale ileum ingesamel is (ileale verteringsmetode). In die derde studie is die produksie prestasie van braaikuikens op ‘n gebalanseerde metaboliseerbare energie (AME) rantsoen, soos beïnvloed deur die verskillende behandelings, geëvalueer. In die eerste studie is knoffel en ‘n kommersiële prebiotikum (Bio-Mos®) geëvalueer en met ‘n antibiotikum (doksisiklien, Doxyveto-SOS) in terme van AND en AAD vergelyk. Beginner- en afrondingsrantsoene is as ‘n meel of pille gevoer. Die knoffel is teen 8g/kg, 13g/kg en 18g/kg in die rantsoen ingesluit. Bio-Mos® is teen 1g/kg, 2g/kg en 3g/kg in die beginner rantsoen en teen 0.5g/kg, 1g/kg en 1.5g/kg in die afrondingsrantsoen, ingesluit. Die antibiotikum is teen 0.3g/kg in beide rantsoene ingesluit. Geen van die behandelings het enige positiewe invloed op AND gehad nie. Deur ‘n verpilde rantsoen te voer het sekere negatiewe invloed op AND gehad. Oor die algemeen het geen behandelings enige positiewe invloed op AAD gehad nie. Op dag 21 en 35 het die insluiting van knoffel teen 18g/kg in ’n meel rantsoen ’n negatiewe invloed op AAD gehad, wanneer dit met die negatiewe- en positiewe kontroles vergelyk is. Dit blyk nie dat om ‘n pil of meel te voer enige invloed op die invloede van die verskillende behandelings gehad het nie. In die tweede studie is Bio-Mos®, ‘n organiese suur (Acid-Pak 4-way®) en ‘n medium-ketting trigliseried (MCT) geëvalueer en met ‘n antbiotikum, doksisiklien (Doxyveto-SOS) in terme van AND en AAD, vergelyk. Beginner- en afrondingsrantsoene is gevoer as ‘n meel. Bio-Mos® is teen 1g/kg, 2g/kg, and 3g/kg in die beginner rantsoen en teen 0.5g/kg, 1g/kg, 1.5g/kg in die afrondingsrantsoen, ingesluit. Acid-Pak 4-way® is teen 0.4g/kg, 1g/kg en 1.6g/kg vir die beginner –en afrondingsrantsoene ingesluit. Die MCT is teen 3g/kg, 3.6g/kg, 4.2g/kg in die beginner rantsoen en teen 2.1g/kg, 2.7g/kg en 3.4g/kg in die afrondingsrantsoen ingesluit. Die antibiotikum is ingesluit teen 0.3g/kg. Geen behandelings het enige betekenisvolle invloed in terme van AND gehad nie. Op dag 21 het MCT (3.4g/kg), in vergelyking met die negatiewe- en positiewe kontrole, die grootste positiewe invloed op AAD gehad. Acid-Pak 4-way® (1g/kg) het, in vergelyking met die positiewe kontrole, ‘n positiewe invloed gehad op AAD. Op dag 28, het Bio-Mos® (0.5g/kg en 1.5g/kg) en Acid-Pak 4-way® (0.4g/kg) die grootste positiewe invloed op AAD gehad. Die behandeling met MCT (2.7g/kg) het die mees negatiewe invloed op AAD gehad. In die derde studie is die insluiting van Bio-Mos®, Acid-Pak 4-way® en MCT getoets om die invloed op braaikuiken produksie prestasie te evalueer, en te vergelyk met die invloed van die insluiting of afwesigheid van ‘n antibiotikum. Liggaamsmassa (BW), liggaamsmassa toename (BWG), voerinname (FI) en voeromsetverhouding (FCR) is gemeet. Beginner- en afrondings rantsoene is gevoer as ‘n meel. Bio-Mos®, MCT en Acid-Pak 4-way® is onderskeidelik teen 3.0g/kg, 4,2g/kg en 1.6g/kg in die rantsoen ingesluit. Die kuikens is met aankoms (per hok) geweeg, asook op dae 7, 14, 21, 28, 35. Voerinname per hok is gemeet op dae 7, 14, 21, 28 en 35. Mortaliteite is daagliks aangeteken. Die insluiting van Acid-Pak 4-way® het in vergelyking met die negatiewe kontrole, Bio-Mos® en MCT insluiting ‘n hoër BWG tot gevolg gehad. Die gevolgtrekking wat gemaak kan word is dat Bio-Mos®, Acid-Pak 4-way® en MCT gebruik kan word as ‘n moontlike alternatief vir antibiotika insluiting. Hierdie drie behandelings was nie noodwending meer effektief as die antibiotika nie, maar het wel bewys dat dit kompeterende alternatiewe is.

碩士
中華科技大學
健康科技研究所在職專班
100
Abstract Cancer was the biggest reason that caused death in Taiwan for many years, furthermore, the global market in cancer had had a high value recently. In 2012, the global immune cancer therapy market was worth more than 372 billions, thus, many countries had had put effort in this area. However, Chinese herbals’functions were utilized through the metabolism of enteric bacteria and transformed it to benefit human body. Therefore, the most important discussion was to find more spices of bacteria to do transformation that benefit human. This experiment was done and invented Symbiotic hermentation by Chen-ping Company, extracting many kinds of enzymes and mushrooms, such as: Saccharomyces cerevisiae, Lactobacillus acidophilus, Bifidobacterium longum, Ganoderma lucidum and Cordyceps sinensis, and fermented together. After that, they used Symbiotic hermentation, mixede with different concentrations of 【CP Hermentation】to allocate different concentrations of samples and added nine kinds of human cancer cells in each allocated sample. By influencing its living period, they analyzed restrained ability of cancer. The result came out that cancer cells living between 24hrs and 72hrs, treated under 1ul of 【CP Hermentation】had no differences compared with normal groups. On the other hand, under treatment of 1, 5, 10, 25, 50l【CP Hermentation】, the cancer cells, （HepG2）,（MCF-7）,（RH2）,(CE48T),(CAL-27),（PC-3）,（T84）,（HCF8）,had distinct lower living period than normal groups. Especially in (CE48T),(CAL-27),（PC-3）the restrained ratio can be high up to 98.2%、98.1%、97.2%, which were the best effects. The more concentrated the less time period cells tended to live and it had dose dependent relationship. However,72hrs treatment in cancer cells, （ CL1-1 ）,（ HCC827）, the best effect was shown in 25l【CP Hermentation】, similar in 24hrs’ treatment, the 50l【CP Hermentation】appeared feedback inhabitation mechanism. High concentrated【CP Hermentation】treated in （ CL1-1 ）,（ HCC827） had drug resistance. They also wanted to know how Symbiotic hermentation effected on human body due to the acknowledgment that plasticizer and preservatives had significant effects on human body and using purification can get rid of components that caused drug resistance and lowered the risk and possibility of the drug resistance. Keywords: CP Hermentation, cancer retain, cancer cells

碩士
國立成功大學
建築學系
102
Since the industrial revolution,as economic development has shifted to world,environmental impact has become an increasingly serious concern.That impels human realize how important the symbiotic relationship between itself and ecological environment.For the concept of sustainable development subsequently proposed already become a common topic of concern.This study mainly investigate about ecological village,derived from sustainable development.In recent years,the concern and protection about the ecological environment has increased much.The technology of ecological construction also has been improved a lot. Moreover,the project of general village reconstruction has raised the whole quality of the village to ecological village.The purpose of ecological village is not only focused on the improvement of the ecological environment maintenance,for how symbiosis is the key factor.uder the investigate how to reach a balance between execute sustainable development and conserve island village's original style that faces industrial transformation and change of environment through carry out ecological village planning. This study selected Longmen village Port in Huxi Township Penghu County as the base for planning and design.Longmen village grown up in the high fish output and rich natural resources,it's developed a unique agriculture and fisheries industry type and it also become one of the important village in Huxi Township.But as the industrial transformation,Urban development and environmental damage related problem,Longmen village has fade off Past the Bustling to reduce edge area.Through the premise of symbiotic ecological theory,this study inspected Longmen village,the ecological resources,historic Monument,local cultural heritage,field images of the surrounding environment,space evident,and field images of the surrounding environment.This study analyzed the area from the perspective of ecological village planning and developed design strategies as the creation achievements.lastly accomplish Longmen village rebirth of the target.

Cancer is the most widely recognized reason for human deaths globally. Conventional anticancer therapies, including chemotherapy and radiation, are very costly and induce severe side effects on the individual. The discovery of anticancer compounds including dairy-derived peptides may thus be a better alternative for cancer prevention and management. Anticancer peptides exist in the amino acid chain of milk proteins and can be generated during proteolytic activities such as gastrointestinal digestion or food processing including fermentation by lactic acid bacteria (LAB) and probiotics. However, proteolytic capacity of these bacteria is strain specific. The study was conducted to establish proteolytic activity of Lactobacillus (L.) acidophilus (ATCC® 4356™), L. casei (ATCC® 393™) and L. paracasei subsp. paracasei (ATCC® BAA52™) in yogurt. Crude peptides were separated by ultra-high centrifugation and tested for antioxidant and antimutagenic activities. The degree of proteolysis highly correlated with these bioactivities, and its value (11.91 %) for samples containing all the cultures was double that of the control. Liberated peptides showed high radical scavenging activities with 1,1- diphenyl-2-picrylhydrazyl and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), IC50 1.51 and 1.63 mg/ml respectively and strong antimutagenicity (26.35 %). These probiotics enhanced the generation of bioactive peptides in yogurt.

博士
國立屏東科技大學
水產養殖系所
106
The aims of this study were to (1) develop a relevant synbiotic to promote growth perfomance of the white shrimp, Litopenaeus vannamei; (2) discover some new insights into the mechanisms of action how synbiotic can exert growth of L. vannamei by using next generation sequencing (NGS) technology and metabolomic approaches; and (3) confirm whether identifed metabolites assuredly exert the growth of L. vanamei. Firstly, three common prebiotics, including fructooligosaccharide (FOS), galactooligosaccharide (GOS) and mannan oligosaccharide (MOS), were screened for supporting the growth of Lacto. plantarum 7-40, prebiotic score, and abilities to induce bacterial enzyme activities of protease, leu-aminopeptidase, and β-galactosidase. Results showed that growth of Lacto. plantarum 7-40 significantly increased after adding GOS as the sole carbon source. GOS induced the highest activities of protease and β-galactosidase by Lacto. plantarum 7-40. These results indicated that GOS was more suitable for combining with Lacto. plantarum 7-40 as a synbiotic for shrimp than FOS and MOS. For growth performance trial, four experimental diets, including a basal diet with no GOS or Lacto. plantarum 7-40 (control), 0.4% GOS (PRE), 108 CFU Lacto. plantarum 7-40 kg 1 (PRO), and 0.4% GOS in combination with 108 CFU Lacto. plantarum 7-40 kg 1 (SYN), were given to shrimp. At the end of a 60-day feeding trial, shrimp fed the SYN diet had significantly higher weight gain (WG), specific growth rate (SGR), lactic acid bacteria (LAB) and intestinal enzyme activities, including protease, leu-aminopeptidase, and β-galactosidase, compared to the control. We firstly concluded that dietary Lacto. plantarum 7-40 and GOS had a synergistic effect on improving the growth performance, intestinal microbiota, and digestive enzyme activities of L. vannamei. Secondly, to discover new insights into the mechanisms of action, shrimp from the synbiotic and control treatments were sampled for the elucidation of intestinal microbial communities by using the next generation DNA sequencing analyses and the detection of the changes in metabolic profiles in hepatopancreas of shrimp were examined by using 1H nuclear magnetic resonance (NMR)-based metabolomic analyses, then reverse-phase high-performance liquid chromatography (RP-HPLC) and spectrophotometric analyses were used to confirm significantly altered metabolites in the hepatopancreas and plasma of shrimp in all groups. Results showed that 6 phyla, 11 classes, 19 orders, 30 families, 58 genera and 73 species with taxonomical names were assigned. The majority of OTUs were shared between synbiotic and control shrimps composed of 37 OTUs. However, intestinal biodiversity analyses revealed that SYN-fed shrimp had higher species richness, eveness, and Shannon-Weaver index than those of the control shrimp, but no statistical significance. Interestingly, shrimp that were fed SYN diet improved colonization of Lacto. plantarum and reduced prevalences of V. harveryi and Photobacterium damselae in the intestines. In addition, results from the 1H NMR analyses unraveled that 22 hepatopancreas metabolites were matched and identified between the SYN- and control- fed shrimp. Among hepatopancreatic metabolites, inosine monophosphate (IMP), valine, and betaine, significantly increased in the SYN group. Confirmation using RP-HPLC and spectrophotometric methods showed that IMP presented high amounts in the hepatopancreas, but not in the plasma of shrimp; in contrast, valine and betaine were in high concentrations in both the hepatopancreas and plasma. Finally, to confirm whether identifed metabolites assuredly exert the growth of L. vanamei, a confimatory study on the effects of metabolite-supplemented diets on growth of L. vannamei was carried out. Six experimental diets, including 0.1% inosine monophosphate (IMP); 0.5% valine (VAL); 0.5% betaine (BET); 0.1% IMP + 0.5% valine + 0.5% betaine (META); synbiotic (SYN); and the control diet (added nothing) were given to shrimp. After the 60-day feeding regimen, results showed that shrimp fed either IMP- or META-supplemented diets significantly increased WG and SGR than that of shrimp in the BET and control groups. In conclusion, results suggested that GOS and the Lacto. plantarum 7-40 had a synergistic effects on growth performance of L. vannamei. Feeding a synbiotic comprised of 0.4% GOS and Lacto. plantarum 7-40 at 108 CFU kg-1 of feed can alter intestinal microbial communities of shrimp by inducing colonization of Lacto. plantarum in the intestine of shrimp and reducing the prevalence of Vibriosis in the intestine of shrimp. In presence of GOS, Lacto. plantarum 7-40 exhibited higher production levels of intestinal protease, leu-aminopeptidase, β-galactosidase enzymes that play important roles in protein digestibility and sugar digestion. Combination of GOS and Lacto. plantarum 7-40 siginificantly increased crucial metabolites, including IMP, valine, and betaine in the hepatopanreas and plasma of shrimp, that would be the important components in protein and energy metabolism in shrimp, of which IMP, a nucleotide, could be a main factor to improve growth performance of L. vannamei.

碩士
國立陽明大學
腦科學研究所
107
The concept of gut-brain axis and importance of gut microbiota have been raised these years. Dysbiosis of gut microbiota is associated with multiple disorders, including asthma, obesity, depression, autism, Parkinson’s disease and Alzheimer’s disease (AD). AD is a neurodegenerative disease that impairs multiple memory and behavior domains without any effective treatment. Whether microbiota modulation can ameliorate cognitive function in AD remains unknown. In our study, we established prebiotics and synbiotic mixture, TCP and TCP+P, as intervention for AD mouse model. Our results demonstrated that after synbiotic treatment for 2 months, AD mice showed improving spatial memory, and increasing neurogenesis compared to untreated mice. Furthermore, the synbiotic treatment also reduced systemic inflammation, Aβ 42 accumulation and plaque formation. Taken together, our study demonstrated the protective effects of synbiotic mixture in AD mice, suggesting that dietary modulation may be a potential treatment to slow down AD processing.

This project involved the development of a synbiotic dairy product and its use in human feeding trials to quantify the hypocholesterolemic and antihypertensive effects of probiotics on hypercholesterolemic men and women. To this end, eleven strains of Lactobacilli and three strains of Bifidobacteria were evaluated for their acid and bile tolerance, cholesterol removal ability and hypotensive properties in vitro. Our results showed that all studied strains were able to tolerate pH 2.0 for two hours. However, strains of L. acidophilus in general were more acid-resistant than Bifidobacteria with an average viable count more than 107 CFU mL-1 after 2h incubation at pH 2.0. All selected strains could tolerate the bile salts, with greater tolerance towards oxgall than to taurocholic acid. The ACE inhibition was found to be significantly different among the strains ranging from 8.2% to 78.5%. In addition to the above, all strains tested possessed varying degrees of cholesterol removal capabilities from the growth, either via medium via incorporation of cholesterol into the cellular structure, or binding of cholesterol onto cellular membrane and/or bile salt deconjugation. The affinity towards cholesterol was found to be greater in growing cells than resting or dead cells; nevertheless, this study confirmed that the latter cells could be useful as cholesterol-reducing agents in gastrointestinal tract (GIT). Fatty acid composition analysis of the harvested cells revealed that cells grown in the presence of cholesterol contained higher level of saturated fatty acids and lower level of unsaturated fatty acids than those grown in the absence of cholesterol, suggesting that cholesterol from the medium may be incorporated into the cellular membrane of probiotics. The membrane of each probiotic cell was broken up by centrifuging at 4000 × g for 20 min and fatty acid composition of harvested cells was analysed using gas chromatography. The selected strains were also screened for their bile salt deconjugation ability and bile salt hydrolase activity (BSH). Strains of Lactobacillus in general showed a higher bile salt deconjugation ability and liberated a more substantial amount of cholic acid than Bifidobacterium strains. Substrate preference for BSH activity was more towards sodium glycocholate than sodium taurocholate, however, most strains exhibited higher total BSH activity on bile salt mixtures than to when bile salts were used individually. Based on the above results L. acidophilus CSCC 2404 and L. rhamnosus ASCC 1520 were selected to develop a synbiotic yogurt to be used in human trials. The selected probiotics were subsequently screened in the presence of four prebiotics, sorbitol, mannitol, fructooligosaccharide, and fructooligosaccharides-enriched inulin (Synergy 1), to determine the best type and concentration of prebiotic in order to achieve maximum cholesterol removal and ACE-I activity inhibition in vitro and in vivo. Screening using factorial design showed that the combination of 1% (v/v) L. acidophilus CSCC 2404 and L. rhamnosus ASCC 1520 in the presence of 2.0% (w/v) Synergy 1 achieved the best results in synbiotic yogurt. In addition to synbiotics, there has been an increased interest in dietary sources of antioxidant polyphenols in red fruit juices such as pomegranate (Punica granatum L.) due to their strong antioxidant activity (AA). Therefore, to further improve the health-promoting properties of the synbiotic yogurt, different levels of pomegranate juice concentrate (PJC) (10, 12.5, 15 and 20 %) were added to yogurt milk before incubation, and the AA, antihypertensive activities and hypocholesterolemic properties of the synbiotic yogurt was evaluated during refrigerated storage for 21 days. Despite the slight adverse effect of PJC supplementation at 20% level on probiotic numbers, the probiotic population at the end of storage period was still within the acceptable range of colony- forming unit (CFU) to deliver health-promoting properties (> 106 CFU mL-1). The resulting synbiotic yogurt showed increased polyphenol levels by 63%, AA by 94%, and angiotensin-converting-enzyme (ACE) inhibition by 75% compared to the control plain yogurt. The sensory evaluation of the PJC-enriched sybniotic yogurt revealed that by increasing the PJC level, the overall acceptability among the panellists increased significantly (P < 0.05). To provide evidence to support the positive effects of this PJC-supplemented synbiotic yogurt on serum lipid profile and blood pressure (BP), an 8-week parallel, doubleblinded, randomised trial was designed and conducted with 48 male and female volunteers. The volunteers were aged between 30-65 years, with serum total cholesterol (TC) and triglyceride (TG) levels less than 6·2 and 2.3 mmol L-1, respectively, a body mass index (BMI) between 25 and 35 kg/m2, and willing to consuming a daily serving of 200g yogurt. Subjects were assigned to three groups; Group 1 (control) consumed the yogurt samples which only contained 0.02% (w/v) freeze-dried YC-380 (Streptococcus thermophilus 1275 and L. bulgaricus 1842), Group 2 consumed the synbiotic yogurt (Syn) containing 0.02% (w/v) freeze-dried YC-380, 0.5% (v/v) L. acidophilus ASCC 2404, L. rhamnosus ASCC 1520, and 2.0% (w/v) Synergy 1, and Group 3 consumed the same yogurt supplemented with 20% (v/v) PJC (Synp). The yogurts were produced and distributed to the participants on weekly intervals. Fasting blood samples, 3-day dietary records, anthropometric measurements and BP were collected at baseline and at the end of four and eight weeks. Comparison of the mean differences of serum TC, TG, and high-density lipoprotein cholesterol (HDL-C) levels among the three groups confirmed a drop of 9.81% and 18.10 % in TG level and TC: HDL-C ratio (P < 0.05) respectively, from baseline to week eight. However, no significant changes from the baseline were observed in systolic blood pressure (SBP) and diastolic blood pressure (DBP) (P > 0.05). The post hoc test showed that the decrease in serum TC and LDL-C was significant for both the Syn and Synp groups (P < 0.001 for both). Consumption of Synp yogurt resulted in 19% and 23% decrease in serum TC and LDL-C levels, respectively, at the end of the study, whereas in the Syn group, these levels were only reduced by 14% and 17% respectively. Overall, the selected probiotics L. acidophilus CSCC 2404 and L. rhamnosus ASCC 1520 proved to have the ability to tolerate acid and bile conditions and were optimised with 2.0% (w/v) Synergy 1 and 20% (v/v) PJC for maximum cholesterol removal in vitro and in vivo. Therefore, this study indicates the combined effectiveness of probiotics, prebiotics and polyphenols in ameliorating cardiovascular disease risk factors in both women and men.

Consumers are currently concerned about improving their health, and therefore demand foods that are beneficial to overall health. This has caused the rising interest in probiotics, which are live microorganisms which when ingested in sufficient amounts, restore balance in the gastrointestinal tract and consequently improve health. Probiotic bacteria have been incorporated into various food products which are now referred to as functional foods, and represent about 65% of the world‟s food market. Probiotics are sensitive to various environmental factors such as oxygen, moisture, pH and temperature. It is of great importance that probiotics remain viable and alive throughout the stages of processing, storage in food products and during gastrointestinal transit in order for them to confer health benefits. The use of prebiotics and microencapsulation to protect and ensure viability of probiotics has been used in food industries. Challenges faced when using most microencapsulation techniques include the need for a food grade encapsulating material, stability of the probiotic cells during encapsulation processes and storage, the need to minimize negative effects they might have on the organoleptic properties of foods into which they are incorporated. The freeze drying technique, which is known to be suitable for the preservation of probiotic cells, avoids heat induced injuries to cells and also slows down detrimental chemical reactions, was used in the current study to prepare microparticles encapsulating probiotic bifidobacteria. Due to limited reports on the use of lipid based food grade encapsulating materials for the microencapsulation of probiotics, this study explored the use of such materials and developed a lipid based synbiotic material which is expected to protect and improve probiotic viability. A lipid based excipient Vegetal BM 297 ATO and various concentrations of the prebiotic inulin were used to prepare different formulations, followed by an investigation to determine which concentration of inulin resulted in better protection and survival of Bifidobacterium longum LMG 13197 during the freeze drying process. Bifidobacterium longum LMG 13197 was successfully encapsulated in Vegetal using freeze drying method. It was observed that the formulation prepared with 2% (w/v) inulin resulted in better protection of B. longum LMG 13197 during the encapsulation process. Characterization of the microparticles revealed that they contained high numbers of bacterial cells resulting from relatively high encapsulation efficiency. The presence of inulin resulted in microparticles with an acceptable size which is desirable for food applications. These results led to further investigation of the potential of Vegetal-inulin matrix to protect bifidobacteria in simulated gastrointestinal fluids and improve shelf life under different storage conditions. This study demonstrates that the Vegetal-inulin matrix protected B. longum LMG 13197 during transit in the simulated gastric fluid (SGF) and subsequently released the cells in the simulated intestinal fluid (SIF). In comparison with the unencapsulated cells, the number of cells released in SIF was higher, which suggests that the Vegetal-inulin matrix has the potential to release probiotics in the colon for health benefits to be exerted. The shelf life of encapsulated B. longum LMG 13197 powders stored in glass bottles was investigated under two different storage temperatures for 6 weeks. The study demonstrates that although there was a high loss of viable probiotic cells during storage at 25°C, Vegetal-inulin matrix improved survival of probiotics for 3 weeks as opposed to the unencapsulated cells. On the other hand, encapsulation with Vegetal did not offer improved survival of bacteria when compared to the unencapsulated cells at 4°C, but the addition of inulin offered better protection for up to 5 weeks. Therefore, better shelf life of Vegetal-inulin microparticles containing B. longum LMG 13197 can be achieved at 4°C than at 25°C.
Dissertation (MSc)--University of Pretoria, 2015.
tm2015
Microbiology and Plant Pathology
MSc
Unrestricted

A 24-hour fermentation of the optimized synbiotic composed of B. longum and EPR was performed to give a cell-free fermentation broth (S24). S24 was co-cultured with two colon cancer cell lines (Caco-2 and SW620) and normal colon cells (FHC). S24 significantly inhibited cell proliferation for both colon cancer cells but promoted FHC cell growth by 10-25% as shown by MTT and BrdU arrays. Primary DNA damage analysis by alkaline comet assay showed S24 caused DNA damage to a comparable extent as the positive control of 10 mM H2O2 (treated for 1 hour) for both cancer cells. Dynamic analysis on DNA damage-associated DNA repair showed the two colon cancer cells had different response pattern to S24. Flow cytometric analysis showed that both Caco-2 and SW620 when treated with S24 (IC 50=3.66 mM of acetate) were arrested initially at G2/M and subsequently at S phase accompanied with large sub-G1 peaks. Dual staining with PI/AnnexinV further proved the appearance of apoptosis. Live cell imaging analysis on Caco-2 cells treated with S24 showed the following events: mitochondria were rapidly destroyed within the first two-hour treatment, the cells bubbled and the nucleus condensed after the mitochondrial had shrunken, followed by apoptosis.
Despite active research on synbiotic on anti-carcinogenesis of colon cancer by synbiotics, the underlying mechanism still remains unclear. This study investigated a novel synbiotic composed of non-digestible carbohydrates (NDCs) extracted from mushroom sclerotia as prebiotics and Bifidobacteria as probiotics. Preliminary results on incubation of two probiotics ( Bifidobacterium longum and Lactobacillus brevis) and one pathogenic bacterium (Clostridium celatum) separately with 3 NDCs extracted from mushroom sclerotia [Poria cocos (PC), Polyporus rhinocerus (PR) and Pleurotus tuber-regium (PT)] indicated that the growth of B. longum and L. brevis was stimulated more preferentially than C. celatum after 72-hour fermentation. The short-chain fatty acid (SCFA) profile was dominated by acetate (> 98% of total SCFAs) with very little butyrate (< 2.0% of total SCFAs) and the organic matter disappearance (OMD) during fermentation was consistent with the bacterial growth. Among the synbiotic combinations, NDC from PR and B. longum gave the largest amount of acetate (2.47+/-0.232 mmol/g of organic matter disappearance).
Results obtained from human pathway finder RT2 Profiler(TM) PCR Array indicated that S24 could modulate the proliferation of colon cancer cells mainly by various pathways such as cell cycle and DNA damage repair, apoptosis and cell senescence, etc. In SW620 cells, PCR Array of Human Cell Cycle further revealed that the modulated genes mainly belonged to the gene cluster of S phase and DNA replication as well as G2 and G2/M transition. While for Caco-2 cells, the cell-cycle modulated genes mainly belonged to the cluster of G2 and G2/M transition. Immuno-blotting on the pivotal upstream regulators showed that phosphorylation of ATM at Serine 1981 was significantly increased in both cancer cells. Site-specific phosphorylation of pRB was decreased and phosphorylation of Chk1 was increased in both cancer cells while Chk2 were increased in SW620 cells. Cdc25A was phosphorylated at serine17 in both cancer cells. It can be proposed that the blockage of DNA synthesis or DNA damage was due to the down-regulation of some pivotal DNA replication related proteins such as RPA3, PCNA and MCMs, detected by ATM-Chk1/Chk2-Cdc25A pathway. This would cause the prolonged staying of cells at the G1/S checkpoint which further moved on to S phase arrest for SW620 cells. Moreover the sharply up-regulated p21, an important inhibitor of Cdk2 would further hinder the cells passing the G1/S checkpoint in SW620 cells.
The tumor suppressor p53 was detected phosphorylated at various sites in SW620 but not in Caco-2 cells. In SW620 cells, G2/M arrest was caused by the inhibition of CDK1/CDC2 due to increased expression of GADD45A and p21 and phosphorylation of Cdc25A, while for Caco-2, the G2/M arrest was caused by degradation of Cdc25A due to the absence of p53-activated GADD45A and p21 expression as shown in the pathway finder results. Some apoptosis-related proteins of Bax, Apaf-1 and PARP were modulated as shown by immuno-blotting in both colon cancer cells. (Abstract shortened by UMI.)
Gao, Shane.
Adviser: Peter Chi-Keung Cheung.
Source: Dissertation Abstracts International, Volume: 72-11, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2009.
Includes bibliographical references (leaves 55-94).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.

O ser humano, integra em si, um reservatório único e bastante diversificado de microrganismos, os quais constituem o microbioma humano. Desequilíbrios no microbioma humano, ou disbiose, estão na origem de diversas doenças, de diferentes etiologias. Os probióticos e prebióticos, são essencialmente prescritos, para prevenção e tratamento de distúrbios gastrointestinais, no entanto, surgem cada vez mais evidências clínicas que provam que os efeitos destes compostos se estendem para além do intestino. Saccharomyces cerevisiae var. boulardii, e as estirpes bacterianas, pertencentes aos géneros Lactobacillus spp. e Bifidobacterium spp., são os microrganismos probióticos mais utilizados em seres humanos. Já os prebióticos mais comuns, são os frutooligossacarídeos, galacto-oligossacarídeos e a inulina. Este trabalho tem como objetivos uma revisão bibliográfica atual sobre as características dos probióticos e prebióticos, os seus benefícios e potenciais mecanismos de ação, na saúde do ser humano, e em diferentes estados de disbiose nos vários sistemas que compõem o corpo humano: no sistema gastrointestinal, no sistema respiratório, no sistema tegumentar, no sistema nervoso central e no sistema cardiovascular. Este estudo permitiu comprovar, um crescimento exponencial no interesse da aplicação dos probióticos e prebióticos, como potenciais agentes terapêuticos em vários estados de disbiose que visem o pleno equilíbrio de saúde no ser humano. Contudo, é crucial a continuidade da investigação científica sobre os seus benefícios no ser humano.
The human being, integrates in itself, a unique and quite diversified reservoir of microorganisms, which constitute the human microbiome. Imbalances in the human microbiome, or dysbiosis, are at the origin of several diseases, of different etiologies. Probiotics and prebiotics are primarily prescribed for the prevention and treatment of gastrointestinal disorders, however, there’s increasing clinical evidence which prove that the effects of these compounds extend beyond the intestine. Saccharomyces cerevisiae var. boulardii, and bacterial strains belonging to the genus Lactobacillus spp. and Bifidobacterium spp. are the most used probiotic microorganisms in humans. The most common prebiotics are Fructo-oligosaccharides, Galactooligosaccharides and inulin. This dissertation aims to provide a current literature review on the characteristics of probiotics and prebiotics, their benefits and potencial mechanisms of action, in human health, and in different states of dysbiosis in the various systems that make up the human body: gastrointestinal system, respiratory system, integumentary system, central nervous system and cardiovascular system. This study showed an exponential growth in the interest in the aplication of probiotics and prebiotics as potencial therapeutic agentes in various states of dysbiosis that aim at achieving full health balance in humans beings. However, it is crucial to continue scientific research on it’s benefits in humans.

Nutrition of children of each age has its particular aspects. For the new born and infantile age it is mainly breast-feeding that is very significant. Nutrition with breast milk is ideally balanced, ensures the intake of high-quality nutrition that is well tolerated by the young organism and ensures the optimal growth and psychological development of the child. Thanks to its composition, breast milk increases the protective impact on the child with respect to infections and creates a natural bond between the mother and the child. In the Thesis, I was trying to look up decisive moments in the infantile nutrition concept both in this country and worldwide and to map the issue of breast feeding and formula feeding since the end of the Second World War up to the present. Also, I wanted to emphasize the irreplaceability of breast milk as wel as supplementing of artificial milks with probiotics, prebiotics, synbiotics and their importance in the nutrition of children. Objective No. 1 of the Thesis has been to look up the key points in the infantile nutrition concept in the world since the beginning of the nineteenth century and since 1945 until this day in the Czech Republic. Objective No. 2 has been to prove that infantile formula nutrition has been focusing also on the immunological aspect apart from the nutritional ones. Objective No. 3 has been to create and aducational program proposal, which can serve as a practical guide for the application of theoretical data in clinical practice. Data necessary to achieve the determined objectives have been acquired by studing, analyses and comparison of available written documents relating to the natural and formula nutrition of infants. Most of these documents have been intended for the educational purposes at secondary school and universities in the Czech Republic. Also, data heve been taken from publications for parents and broad public published since 1945 until this day. By way of illustration, certain charts containing schemes of artificial nutrition corresponding to the time of creation have been used. On these schemes I wanted to show the chages in the concepts brought about by the development not only in the knowledge and thinking of people dealing with the nutrition of infants but also in the new possibilities and technology of the food processing industry.

碩士
臺北醫學大學
保健營養學系
93
This study was to investigate the effects of synbiotics on the intestinal function and antioxidative status in rats. The male Sprague-Dawley rats were randomly divided into three groups such as control (C), low (L) and high dosage (H) groups (1.56 and 7.80g/kg BW/day synbiotics powder was added to diet). The synbiotics was composed of inulin and 5 probiotics (FloraGuardâ, Viva Life Science). After 8 weeks, rats in L group showed the significantly lower fecal E. coli count than that of control group. The jejunal lactase activity and erythrocytic GRd activity were significantly increased in L group. Rats in H group had higher fecal Lactobacillus and Bifidobacterium and lower E. coli. counts than those of C group. H group also showed the significant increase in jejunal lipase, lactase and isomaltase activities. In addition, the jejunal GRd, CAT and SOD activities, erythrocytic GRd and SOD activities, and hepatic CAT activity were significantly increased in H group.

The purpose of the study was to compare the impact of 3 weeks of consumption of commercial yoghurt enriched with Bifidobacterium animalis subsp. lactis BB-12 and Lactobacillus acidophilus LA-5 (P), and the same yoghurt supplemented with prebiotic-inulin (S), on feacal bacterial counts of healthy adults. A randomized, parallel-group, crossover, open-label – DBPC intervention with 66 volunteers was carried out. The 3 week administration of both P and S yoghurt increased (P lower 0.01) counts of bifidobacteria and Lactobacillus acidophilus, and reduced (P lower 0.01) counts of Clostridium spp., Enterococcus spp. and E. coli in fecal samples, in comparison with the control croup. The consumption of probiotics and synbiotics yoghurt reduced (P lower 0.05) the counts of tyramine producing enterococci and E. coli among the faecal isolates. In the synbiotics group, counts of bifidobacteria and Lactobacillus acidophilus remained higher, and counts of Clostridium spp., Enterococcus spp. and E. coli lower (P lower 0.05) after 1 week wash out period, in comparison with the preadministration phase. It was concluded that the synbiotics yoghurt was superior to the probiotics product in maintaining the increased levels of beneficial bacteria and decreased counts of potentially patogens bacteria in the intestine.