Academic literature on the topic 'Bifidobacterium animalis subsp lactis Bb12'

ABSTRACT A real-time PCR method has been developed to distinguish Bifidobacterium animalis subspecies in the gastrointestinal tracts of pigs. Identification of a highly conserved single-copy tuf gene encoding the elongation factor Tu involved in bacterial protein biosynthesis was used as a marker to differentiate homologous Bifidobacterium animalis subsp. lactis (strain Bb12) from Bifidobacterium animalis subsp. animalis, as well as Bifidobacterium suis, Bifidobacterium breve, Bifidobacterium longum, several species of Lactobacillus, and Enterococcus faecium. Real-time PCR detection of serially diluted DNA extracted from a pure culture of Bb12 was linear for bacterial numbers ranging from 10 to 10,000 tuf gene copies per PCR (r 2 = 0.99). Relative differences in Bb12 bacterial numbers in pigs fed daily with Bb12 were determined after detection of Bb12 tuf gene copies in DNA extracted from the intestinal contents. Piglets treated with Bb12 immediately after birth maintained a high level of Bb12 in their large intestines with continuous daily administration of Bb12. Piglets born to Bb12-treated sows during the last third of their gestation and also treated with Bb12 at birth (T/T group) had a higher number of Bb12 organisms per gram of intestinal contents compared to placebo-treated piglets born to placebo-treated sows (C/C group), Bb12-treated sows (T/C group), or piglets born to placebo sows but treated with Bb12 immediately after birth (C/T group). In addition, there was a significant increase in gene expression for Toll-like receptor 9 (TLR9) in piglets from the T/T group, with no change in TLR2 and TLR4. These findings suggest that the tuf gene represents a specific and functional marker for detecting Bifidobacterium animalis subsp. lactis strain Bb12 within the microbiota of the intestine.

The commercial probiotic strain Bifidobacterium animalis subsp. lactis Bb12 was encapsulated using emulsion encapsulation into milk protein matrix without and with the addition of 0.5% w/w lecithin into the oil. Different agitation speeds were used during the encapsulation process. The examination of microcapsules was carried out by optical microscope and fluorescence in situ hybridisation. The particle size distribution as volume based median d_0.5 was evaluated by the laser diffraction method. In the case of no lecithin addition, the agitation speed did not influence significantly the size of the microcapsules. The addition of 0.5% (w/w) of lecithin into the oil caused a decrease of d_0.5value from 196 ± 37 µm to 79 ± 3 µm at an agitation speed of 500 rpm, and from 193 ± 24 µm to 39 ± 3 µm at 1200 rpm. It can improve the sensory properties of the products with the added microcapsules.  

Eight types of capsules containing Bifidobacterium animalis subsp. lactis Bb12 with addition of inulin and/or ascorbic acid were prepared by emulsion method with milk protein matrix or by extrusion method with alginate matrix. The size of protein and alginate capsules containing only Bb12 was 204 ± 18 µm and 1.7 ± 0.1 mm, respectively. Addition of both inulin (1% w/w) and ascorbic acid (0.5% w/w) increased the size of alginate capsules. Both methods of encapsulation prevented efficiently the manifestation of Bb12 cell metabolic activity. All types of encapsulation provided higher resistance of Bb12 cells to the conditions of a model gastrointestinal tract (GIT) compared to free cells. The influence of co-encapsulation with inulin (1% w/w) and ascorbic acid (0.5% w/w) on viability in model GIT was not demonstrable in alginate capsules but it was significant in protein capsules. The most efficient was co-encapsulation in a protein matrix with 1% w/w inulin and 0.5% w/w ascorbic acid.<br /><br />

The cells of commercial strain Bifidobacterium animalis subsp. lactis Bb12 were encapsulated using emulsion encapsulation in a milk protein matrix. The volume based median of the microcapsules was 52.1 ± 6.2 µm. The stability of free and encapsulated cells was compared during 28 day-storage in pineapple juice and in strawberry-apple juice at 8 ± 1°C and 22 ± 1°C. Encapsulation ensured a higher number of cells compared to the free cells only at 8 ± 1°C. Strawberry-apple juice was found to be not suitable as probiotic vehicle. Both free and encapsulated cells lost their viability after 14 days at 22 ± 1°C. The number of bifidobacteria cells, pH and lactic and acetic acid content did not change in pineapple and strawberry-apple juice after 24 h cultivation at 37°C.

The aim of the present study is to background the effect of conjugated lino-leic acid isomers (CLA) produced by two probiotic strains, Lactobacillus rham-nosus LBRE-LSAS (a human originated bacterium) and Bifidobacterium ani-malis subsp lactis Bb12, on both hepatic and adipose tissues of high-fat diet fed Wistar rats. Five-week-old male Wistar rats were divided into 4 groups (n=6) fed a high-fat diet for three of them (control and supplemented with 1x109 CFU per rat of LBRE-LSAS or Bb12 strain and 1.4% of free linoleic acid; designed as treated rats) and a standard diet for the fourth group. After 8 weeks of experimental period, rats were sacrificed after chloroform anesthe-sia; livers and adipose tissues of each group were excised for biochemical and histological analyses. Obtained results showed that livers of treated high-fat diet fed rats did not exhibit a hepatic steatosis like those of untreated high-fat diet fed rats (control group) did. Lipid profile (triglycerides and total cholesterol) of the liver and the adipose tissue was markedly improved in treated rat groups, especially in LBRE-LSAS strain given high-fat diet rats. Such results strongly support the occurrence of the bacterial power of Lacto-bacillus rhamnosus LBRE-LSAS and Bifidobacterium animalis subsp lactis Bb12 to modulate lipid metabolism and to avoid steatosis in diet-induced model of obesity in rat.

Preterm germ-free piglets were monoassociated with probiotic Bifidobacterium animalis subsp. lactis BB-12 (BB12) to verify its safety and to investigate possible protection against subsequent infection with Salmonella Typhimurium strain LT2 (LT2). Clinical signs of salmonellosis, bacterial colonization in the intestine, bacterial translocation to mesenteric lymph nodes (MLN), blood, liver, spleen, and lungs, histopathological changes in the ileum, claudin-1 and occludin mRNA expression in the ileum and colon, intestinal and plasma concentrations of IL-8, TNF-α, and IL-10 were evaluated. Both BB12 and LT2 colonized the intestine of the monoassociated piglets. BB12 did not translocate in the BB12-monoassociated piglets. BB12 was detected in some cases in the MLN of piglets, consequently infected with LT2, but reduced LT2 counts in the ileum and liver of these piglets. LT2 damaged the luminal structure of the ileum, but a previous association with BB12 mildly alleviated these changes. LT2 infection upregulated claudin-1 mRNA in the ileum and colon and downregulated occludin mRNA in the colon. Infection with LT2 increased levels of IL-8, TNF-α, and IL-10 in the intestine and plasma, and BB12 mildly downregulated them compared to LT2 alone. Despite reductions in bacterial translocation and inflammatory cytokines, clinical signs of LT2 infection were not significantly affected by the probiotic BB12. Thus, we hypothesize that multistrain bacterial colonization of preterm gnotobiotic piglets may be needed to enhance the protective effect against the infection with S. Typhimurium LT2.

Background: Probiotics may prevent the allergic response development due to their anti-inflammatory and immunomodulatory effects. The aim of this study is to determine if the prophylactic treatment with a mixture of Bifidobacterium animalis subsp. Lactis BB12 and Enterococcus faecium L3 would reduce symptoms and need for drug use in children with allergic rhinitis (AR). Methods: The study included 250 children aged from 6 to 17 years, affected by AR. Patients were randomly assigned to the intervention group (150) or to the placebo group (100). Patients in the intervention group, in addition to conventional therapy (local corticosteroids and/or oral antihistamines), were treated in the 3 months preceding the onset of symptoms related to the presence of the allergen to which the children were most sensitized, with a daily oral administration of a probiotic mixture containing the Bifidobacterium animalis subsp. Lactis BB12 DSM 15954 and the Enterococcus faecium L3 LMG P-27496 strain. We used Nasal Symptoms Score (NSS) to evaluate AR severity before and after the treatment with probiotics or placebo. Results: the patients in the intervention group had a significant reduction in their NSS after probiotic treatment (p-value = 2.2 × 10−10. Moreover, for the same group of patients, we obtained a significant reduction in the intake of pharmacological therapy. In particular, we obtained a reduction in the use of oral antihistamines (p-value = 2.2 × 10−16), local corticosteroids (p-value = 2.2 × 10−13), and of both drugs (p-value 1.5 × 10−15). Conclusions: When administered as a prophylactic treatment, a mixture of BB12 and L3 statistically decreased signs and symptoms of AR and reduced significantly the need of conventional therapy.

The market for new functional foods and food supplements is rapidly evolving, with a current emphasis on using natural sources. Algae, probiotics, and colostrum are rich sources of nutrients and bioactive compounds with positive effects on human and animal health. To determine the potential for developing new functional foods combining these components, we evaluated their synergistic effects. We assessed the growth of selected bifidobacteria in a medium supplemented with Chlorella vulgaris and its immunomodulatory and cytotoxic effects on the human peripheral mononuclear cells and colon cancer cell lines Caco-2 and HT29. The hypocholesterolemic effects of Chlorella powder and bovine colostrum fermented by Bifidobacterium animalis subsp. lactis BB12® on lipid metabolism in rats fed a high-fat diet were also determined. Chlorella addition promoted Bifidobacteria growth, with significantly increased inflammatory cytokine (TNF-α and IL-6) levels following 1.0% (w/v) Chlorella stimulation. Rats fed diets containing fermented colostrum with 0.5% (w/v) added Chlorella powder exhibited significantly decreased triglyceride, very low-density lipoprotein, and alanine and aspartate aminotransferase levels, compared to those of the control group. These results support that C. vulgaris is not cytotoxic in intestinal cell models and affords prebiotic and immunomodulatory effects, as well as synergistic triglyceride-lowering effects with bovine colostrum and B. animalis subsp. lactis BB-12.

Adhesion to intestinal mucus is the first event in the process by which intestinal microbes colonize the intestine. It plays a critical role in the initiation of interactions between gut microbes and host animals. Despite the importance, the adhesion properties of probiotics are generally characterized using porcine mucin; adhesion to human mucus has been poorly characterized. In the present study, human intestinal mucus samples were isolated from 114 fecal samples collected from healthy infants and adults. In initial screening, four out of the 13 beneficial microbes tested, including the type strain of Bifidobacterium bifidum, B. bifidum TMC3115, Lacticaseibacillus rhamnosus GG, and Bifidobacterium animalis subsp. lactis Bb12, showed strong adhesion abilities to human mucus. The type strain of B. bifidum and TMC3115 adhered more strongly to neonatal and infant mucus than to adult mucus, while L. rhamnosus GG and B. lactis Bb12 adhered more strongly to adult mucus than to infant mucus. Similar results were obtained for ten additional strains of B. bifidum. In conclusion, age/generation-related differences were observed in the adhesion properties of B. bifidum and other strains. A deeper symbiotic relationship may exist between infants, particularly neonates, and B. bifidum based on its enhanced adhesion to neonatal intestinal mucus.

Probiotics are living organisms which exert a beneficial health effect when consumed in sufficient numbers. Consumer interest in probiotics has increased dramatically in recent years prompting an increase in production and development of functional foods. One major problem is the decreased viability of probiotic bacteria during functional food production and storage and subsequent digestion due to environmental stresses. The most common probiotic strains belong to the genus Lactobacillus or Bifidobacterium. Due to the anaerobic nature of these bacteria, they lack the required defense mechanisms for oxidative stress inherent in aerobic microorganisms. This study examined the oxidative stress responses of six strains of Bifidobacterium, which are commonly used as probiotics in functional foods.The first phase of the study investigated the innate and inducible hydrogen peroxide (H2O2) stress response of Bifidobacterium longum strains NCC2705 and D2957, Bifidobacterium longum ssp. infantis ATCC 15697, and Bifidobacterium animalis ssp. lactis strains BL-04, DSM10140 and RH-1. Strains were screened for survival at increasing concentrations of H2O2 and lethal and sublethal concentrations were determined for each. In the second phase, B. animalis ssp. lactis strains BL-04 and DSM10140 and B. longum strains NCC2705 and D2957 were treated with a sublethal H2O2 concentration and RNA samples were collected for transcriptome analysis after 5 min and either 20 or 60 min. Statistical analysis was performed to identify genes that increased or decreased in expression during H2O2 treatment compared to control cells.Results showed that survival was species and strain dependent and that strains which naturally survived higher H2O2 concentrations had a larger number of differentially expressed genes early on during H2O2 exposure. Some of the protective genetic systems that were activated during H2O2 stress are mechanisms which perform basic cellular functions under normal conditions such as deoxuynucleotide synthesis. Under stress conditions, these systems can be used to detoxify oxidative free radicals. Also a number of genes involved in sugar transport and energy production for the cell showed increased expression, which reveals the increased energy needs of the cells during oxidative stress.During testing, it was found that two B. animalis ssp. lactis strains, BL-04 and DSM10140, had differing levels of survival and gene expression during H2O2 exposure despite having almost identical genome sequences. It was determined that one possible cause of the differences was a genetic deletion in a gene that allows the cell to incorporate extracellular fatty acids into the cell membrane instead of synthesizing them.Results from this project have increased the understanding of oxidative stress responses in bifidobacteria and highlighted possible methods to increase bacterial survival during food manufacture, storage, and human digestion.

Os objetivos do estudo foram elaborar iogurtes com a cultura tradicional e a cultura probiótica de Bifidobacterium animalis subsp. lactis, desidratar os produtos em spray dryer utilizando maltodextrina como carreador e caracterizar os pós obtidos, bem como determinar a resistência dos probióticos ao processo de atomização. O presente estudo avaliou três temperaturas de entrada do ar de secagem (130, 150 e 170°C) em iogurtes com duas diferentes concentrações de maltodextrina (10 e 20%), totalizando 6 tratamentos: T1 (10%malto/130°C), T2 (20%malto/130°C), T3 (10%malto/150°C), T4 (20%malto/150°C), T5 (10%malto/170°C) e T6 (20%malto/170°C). A secagem do iogurte foi realizada em spray dryer piloto e a enumeração das células viáveis de Bifidobacterium animalis subsp. lactis foi realizada através de plaqueamento em profundidade. Os pós apresentaram baixos valores de umidade e elevada higroscopicidade. A atividade de água (Aw) dos pós variou de 0,09 a 0,19 e aumentou após 30 dias de armazenamento, comprovando o caráter higroscópico dos pós obtidos. Verificou-se que após a desidratação dos iogurtes, apesar deles apresentarem contagens inferiores que os produtos integrais, ainda apresentaram contagens acima de 106 UFC/g, ou seja, ainda estavam dentro do limite estabelecido pela legislação para um produto ser considerado probiótico. Os tratamentos que passaram por maiores temperaturas durante o processamento de secagem (T5 e T6) foram os que tiveram maiores perdas de micro-organismos probióticos, sugerindo que as altas temperaturas exerceram forte influência na viabilidade dos probióticos. O T1 obteve maiores contagens do micro-organismo analisado, com contagens acima de 106 UFC/g, com até 60 dias de armazenamento, indicando ser o melhor tratamento entre os estudados, em relação à obtenção de um iogurte caprino probiótico em pó com maior período de vida de prateleira. De maneira geral, conclui-se que o processo de atomização possibilitou a obtenção de iogurte de leite de cabra em pó estável do ponto de vista microbiológico. Além disso, obteve-se um produto que pode ser uma alternativa para incrementar o consumo de leite de cabra, bem como o de probióticos.
The aim of this study was to develop yogurts with the traditional culture and Bifidobacterium animalis subsp. lactis probiotic culture, dehydrate products in spray drying using maltodextrin as a carrier and characterize the powders, as well as determining the resistance of probiotics to atomization process. The present study evaluated three different inlet air temperatures of spray dryer (130, 150 and 170°C) in yoghurts with two different maltodextrin concentrations (10 and 20%), totaling six treatments: T1 (10%malto/130°C), T2 (20%malto/130°C), T3 (10%malto/150°C), T4 (20%malto/150°C), T5 (10%malto/170°C) e T6 (20%malto/170°C). The yogurt drying was performed in a pilot spray dryer and the viable cells of Bifidobacterium animalis subsp. lactis enumeration was performed by pour plate. The powders showed low levels of humidity and high hygroscopicity. The water activity (Aw) of the powders ranged from 0.09 to 0.19 and increased after 30 days of storage, showing the hygroscopic powders character. It was found that after yogurt dehydration, despite their counts were less than integral products, still had counts above 106 CFU/g, therefore were still within regulation limits for a product to be considered as probiotic. The treatments that have undergone higher temperatures during the drying process (T5 and T6) were those who had higher losses of probiotic microorganisms, suggesting that high temperatures had a strong influence on the viability of probiotics. The T1 (130°C/10%) obtained higher counts of the microorganism analyzed, with counts above 106 CFU/g, during 60 days of storage, indicating that is the best treatment among those studied in relation to obtaining a goat probiotic yoghurt powder with longer shelf life. In general, it is concluded that the atomization process allows the obtention of stable goat milk yogurt powder, in a microbiological point of view. Furthermore, it was obtained a product that can be an alternative for increasing the consumption of goat milk as well as probiotics.

O objetivo deste trabalho foi estudar a viabilidade de Bifidobacterium animalis subsp. lactis HN019 em fórmulas infantis fermentadas ou não, probióticas durante armazenamento a 4°C. Três matrizes lácteas e três não lácteas (a base de soja) foram utilizadas para a elaboração de produtos fermentados ou não fermentados usando Bifidobacterium animalis subsp. lactis HN019, resultando em doze diferentes fórmulas probióticas para lactentes. O perfil de acidificação foi determinado a 42°C até pH 4,7. Determinações físico-químicas (sólidos totais, proteína, gordura, cinzas, carboidratos, calorias, densidade e pH) foram realizadas e foram focadas as contagens de bactérias viáveis durante o armazenamento refrigerado. A caracterização química dos produtos lácteos e a não lácteos apresentou resultados diferentes, à exceção FSL2, todos estavam de acordo com Codex Alimentarius. O perfil de acidificação de Bifidobacterium animalis subsp. lactis HN019 diferiu conforme a matriz. Durante o armazenamento dos produtos a 4°C, a contagem de bactérias viáveis de acordo com o preconizado, bem como a pós-acidificação, estando em conformidade com as recomendações da legislação brasileira. Processo (fermentação ou adição) e tipo de matriz (lácteos e não lácteos) influenciaram a pós-acidificação e a viabilidade de Bifidobacterium animalis subsp. lactis HN019. As fórmulas para lactentes podem ser considerados bons veículos de Bifidobacterium animalis subsp. lactis HN019.
This study proposed to study infant formulas as vehicles for Bifidobacterium animalis ssp.lactis HNOI9. Three dairy and three non-dairy matrices were employed for the preparation of fermented or unfermented products using Bifidobacterium animalis ssp. lactis HN019 resulting in twelve different probiotic infant formulas. Acidification profile of the probiotic was determined at 42°C until pH 4.7. Physicochemical determination (total solids, protein, fat, ash, carbohydrates and calories, density and pH) was conducted, and counts viable bacteria (in dairy and non dairy infant formulas fermented and unfermented) during cold storage was focused on. The chemical characterization of the dairy and non-dairy matrix showed different results, the exception FSL2, all were in accordance to the Codex Alimentarius. The acidification profile of B. animalis ssp. lactis HN019 differed according to the matrix. During storage of products at 4°C counts of viable bacteria were stable as well as post-acidification, and were in accordance with the recommendations of the Brazilian legislation. Process (fermentation or addition) and matrix type (dairy and non-dairy) influenced post-acidification and viability of B. animalis ssp. lactis BN019 . Infant formulas could be considered good vehicles for Bifidobacterium animalis ssp. lactis HN019.

Probióticos do gênero Lactobacillus estão sendo amplamente investigados no tratamento da periodontite. Contudo, os efeitos de microrganismos do gênero Bifidobacterium ainda são pouco conhecidos. Este estudo avaliou os efeitos do probiótico Bifidobacterium animalis subsp. lactis (B. lactis) HN019 como adjuvante à raspagem e alisamento radicular (RAR) no tratamento da periodontite experimental (PE) em ratos. No dia 0 do experimento, 32 ratos foram alocados em 4 grupos: C (controle), PROB (probiótico), PE-RAR e PE-RARPROB. Nos grupos PE-RAR e PE-RAR-PROB, a PE foi induzida pela colocação de ligaduras de seda ao redor dos primeiros molares inferiores dos animais. No 14° dia, as ligaduras foram removidas e realizou-se a RAR. Nos animais dos grupos PROB e PE-RARPROB, o probiótico B. lactis HN019 foi administrado diariamente (10 mL/dia de 109 unidades formadoras de colônia) por 15 dias tendo seu início no 14° dia do experimento. Os animais de todos os grupos foram submetidos à eutanásia 29 dias após o início do experimento. As hemimandíbulas e amostras de intestino delgado foram coletadas. Foram realizadas análises histomorfométricas, microtomográficas e imunohistoquímicas. Foram investigados, também, os efeitos microbiológicos de B. lactis HN019 no biofilme associado às ligaduras durante o desenvolvimento da PE. Todos os dados foram analisados estatisticamente. O Grupo PE-RAR-PROB apresentou menores reabsorção óssea alveolar e perda de inserção conjuntiva quando comparado ao Grupo PE-RAR, bem como menor número de osteoclastos, maior expressão de citocinas anti-inflamatórias e menor expressão de citocinas pró-inflamatórias (p <0,05). No grupo PE-RAR-PROB, os valores médios da profundidade da cripta do jejuno e duodeno foram significativamente maiores que aqueles do grupo PE-RAR. A proporção de bactérias aeróbias/anaeróbias foi maior nas amostras de biofilme de animais tratados com B. lactis HN019 em relação àquelas de animais não tratados (p <0,05). Dentro dos limites deste estudo, pode-se concluir que a utilização de B. lactis HN019 como adjuvante à RAR promove benefícios histológicos, microtomográficos e imunológicos adicionais no tratamento da PE em ratos, bem como melhora a morfologia intestinal.
Lactobacillus probiotics have been investigated in periodontitis. However, the effects of the genus Bifidobacterium on periodontitis are hardly known. This study evaluated the effects of the probiotic Bifidobacterium animalis subsp. lactis (B. lactis) HN019 as an adjunct to scaling and root planing (SRP) in rats with experimental periodontitis (EP). At baseline, 32 rats were assigned to 4 groups: C (control), PROB, EP-SRP and EPSRP- PROB. In groups EP-SRP and EP-SRP-PROB, the mandibular first molars of the animals received a ligature. At day 14, the ligatures were removed and SRP was performed. Animals of groups PROB and EP-SRP-PROB were orally administered with 10 mL/day of 109 colony forming units of B. lactis HN019 for 15 days, starting at day 14. Animals were euthanized at day 29. The jaws and samples of the duodenum, jejunum, and ileum were resected. Histomorphometric, microtomographic and immunohistochemical analyses were performed. Microbiological effects of B. lactis on biofilm were also evaluated. Data were statistically analyzed. Group EP-SRP-PROB presented reduced alveolar bone resorption and attachment loss when compared with Group EP-SRP (p<0.05). Group EP-SRP-PROB showed significantly fewer osteoclasts, increased expression of anti-inflammatory cytokines and reduced expression of proinflammatory cytokines compared with Group EP-SRP (p<0.05). In group EP-SRPPROB, the mean values of crypt depth of the jejunum and dudoenum were significantly higher than the ones from group EP-SRP. B. lactis promoted a higher ratio between aerobic and anaerobic bacteria in biofilm samples (p<0.05). Within the limits of this study it can be concluded that the use of B. lactis HN019 as an adjunct to SRP promotes additional histologic, microtomographic and immunologic benefits in the treatment of EP in rats and improves the intestinal morphology.

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This work targetet the caprine ice cream production added with probiotic bacteria Bifidobacterium animalis subsp. lactis. It is divided into two parts. In the first one, four caprine ice cream formulations were evaluated, in which it was used hydrogenated fat (F1 and F3) or fat substitute (F2 and F4) in two different flavors (F1 and F2, passion fruit, F3 and F4, guava). Statistical differences (p<0.05) were detected for their physical-chemical properties, mainly for total solids and fat, but no differences were observed for melting test results. When it went to sensory acceptance, all four ice cream formulations reached high acceptance indexes, mostly formulation F4, which was selected for further studies. In the second part, F4 formulation was prepared with the addition of probiotic bacteria Bifidobacterium animalis subsp. lactis. The growth kinetics was studied and it was observed that the cellular concentration peak was reached after four fermentation hours (10.14 log UFC/g). This time was selected for pre-fermentation procedure and posterior addition at ice cream syrup. In this part of the study, two experimental groups were evaluated: group G1, in which the probiotic addition occurred before the maturation step and group G2, which included a pre-fermentation step and probiotic addition after ice cream maturation. The physical-chemical properties of these two ice cream groups were similar, except for pH, which was higher for group G2 (p<0.05). G1 samples had superior melting rate (3.566 mL/min) and both groups presented microbiological and sanitary results in accordance to current Brazilian legislation. Also, G1 and G2 were considered sensory accepted due to their acceptance indexes higher than 70%. G1 and G2 sensory profiles were similar (p>0.05), and both ice cream samples exhibited high creaminess (6.76 to 6.91) and mouth melting sensation (6.53 to 6.67) scores, while low sandiness scores (0.85 to 0.86) were observed, positive characteristics for this kind of food product. During the first 24 hours after ice cream production, the population of B. animalis subsp. lactis decreased, reaching 7.15 e 6.92 log CFU/g for G1 and G2, respectively. Probiotic bacteria counts fluctuated in ice cream samples during the first 108 days at frozen storage, especially for G2 group. Decreased probiotic viability was observed for G1 samples during the first 35 days of frozen storage, mild variation between 35 and 63 days and stabilized counts were observed after this time. After 21 days at frozen storage, ice cream samples of G1 and G2 groups reached 1.2 x 109 and 1.3 x 109 CFU/portion, respectively. After 108 days under these storage conditions, the survival rate of B. animalis subsp. lactis was 94.26% and 81.10% for G1 and G2 samples, respectively. After simulation of gastroenteric conditions, G2 group reached 9.72 x 105 CFU/portion. Considering the current requirements of Brazilian legislation, which stipulates that functional foods must have minimum probiotic count between 108 and 109 CFU/portion and detectable probiotic bacteria after being submitted to gastroenteric conditions, it is concluded that the ice cream with the addition of Bifidobacterium animalis subsp. lactis made as shown in this work, can be considered as a dairy functional food
O presente trabalho teve o objetivo de estudar a produ??o de sorvete elaborado com leite caprino com adi??o da bact?ria probi?tica Bifidobacterium animalis subsp. lactis. Foi estruturado em duas etapas. Inicialmente foram avaliadas quatro formula??es de sorvete, nas quais foi utilizada gordura vegetal hidrogenada (F1 e F3) e substituto de gordura (F2 e F4) em dois sabores (F1 e F2, maracuj?; F3 e F4, goiaba). Os c?lculos de densidade aparente e overrun foram feitos levando em considera??o as especifica??es da RDC n?. 266 de setembro de 2005. Tamb?m foram realizadas determina??es de pH, acidez total titul?vel e s?lidos totais, s?lidos sol?veis e res?duo por incinera??o, m?todo Kjeldahl para determina??o de prote?na bruta, lip?dios, a??cares redutores e totais. Al?m destas an?lises foram realizados o teste de derretimento e as an?lises microbiol?gicas nos sorvetes elaborados. As formula??es apresentaram diferen?as significativas (p<0,05) quanto ? composi??o f?sico-qu?mica, sobretudo no que diz respeito ao teor de s?lidos totais e gordura, mas n?o foi observada influ?ncia da formula??o sobre o perfil de derretimento das amostras. Quanto ? avalia??o sensorial, as quatro formula??es apresentaram elevados ?ndices de aceita??o, sobretudo a formula??o F4. Essa formula??o foi selecionada para dar prosseguimento aos estudos, cujo objetivo foi estudar os procedimentos de elabora??o do sorvete de leite de cabra com adi??o de B. animalis subsp. lactis. O pico de concentra??o celular (10,14 log UFC/g) foi alcan?ado ap?s quatro horas de cultivo sendo esse ponto escolhido para o procedimento de pr?-fermenta??o e adi??o de B. animalis subsp. lactis na calda do sorvete. Foram avaliados dois grupos experimentais de sorvete caprino com adi??o de probi?ticos: o grupo G1, com adi??o das bifidobact?rias antes da matura??o e G2, com etapa de pr?-fermenta??o e adi??o ap?s a matura??o. As propriedades f?sico-qu?micas dos dois grupos foram similares, com exce??o do pH, cujo valor foi superior no grupo G2 (p<0,05). O grupo G1 apresentou maior (p<0,05) taxa de derretimento (3,566 mL/min) e ambos os tratamentos apresentaram padr?es microbiol?gicos e sanit?rios dentro do exigido pela legisla??o vigente. Os dois grupos foram considerados aceitos sensorialmente, por exibirem n?veis de aceita??o superiores a 70% em todos os atributos verificados. Os perfis sensoriais das amostras G1 e G2 foram semelhantes (p>0,05), com elevados escores para os atributos cremosidade (6,76 a 6,91) e derretimento na boca (6,53 a 6,67), al?m de pontua??o reduzida para o quesito arenosidade (0,85 a 0,86), resultados considerados positivos para esse tipo de alimento. Foi observado decr?scimo da popula??o de B. animalis subsp. lactis ap?s as primeiras 24 horas de produ??o com contagens de 7,15 e 6,92 log UFC/g para G1 e G2, respectivamente. A contagem de bact?rias probi?ticas apresentou varia??es ao longo do armazenamento congelado por 108 dias, sobretudo para o grupo G2. O grupo G1, por sua vez, apresentou queda de viabilidade durante os primeiros 35 dias de congelamento, leve varia??o entre 35 e 63 dias de armazenamento e tend?ncia ? estabiliza??o ap?s esse ponto. Ap?s 21 dias sob armazenamento congelado, as amostras G1 e G2 apresentaram contagem de 1,2 x 109 e 1,3 x 109 UFC/por??o, respectivamente, conforme determina a legisla??o para contagens m?nimas por por??o de comest?veis gelados. Por sua vez, ao final de 108 dias sob essas condi??es, as taxas de sobreviv?ncia do B. animalis subsp. lactis do grupo G1 e G2 foram respectivamente, 94,26% e 81,10%. Ap?s ser submetido a condi??es g?stricas e ent?ricas simuladas in vitro, o sorvete com quatro meses de armazenamento do tratamento G2 apresentou contagem 9,72 x 105 UFC/por??o. Considerando o disposto pela legisla??o nacional vigente, segundo a qual um alimento com alega??o funcional deve possuir contagem m?nima probi?tica entre 108 e 109 UFC/por??o e exist?ncia de microrganismos vi?veis ap?s exposi??o ?s condi??es gastroent?ricas, conclui-se que o sorvete com adi??o de B. animalis subsp. lactis, produzido no presente estudo, constitui alimento l?cteo funcional

O objetivo deste estudo foi avaliar os efeitos da administração tópica de bactérias probióticas do gênero Bifidobacterium na doença periodontal experimental em ratos. Foram utilizados 32 ratos divididos nos seguintes grupos: CT (controle), DPT (doença periodontal), CT-HN019 (controle + probiótico) e DPT-HN019 (doença periodontal + probiótico). No dia 0 do experimento, a doença periodontal foi induzida nos animais dos grupos DPT e DPT-HN019 por meio da colocação de ligaduras ao redor dos primeiros molares inferiores. Nos Grupos CT-HN019 e DPT-HN019, 2 mL de uma suspensão contendo 109 unidades formadoras de colônia/mL de Bifidobacterium animalis subsp lactis (B. lactis) HN019 foram administrados topicamente na região subgengival dos primeiros molares inferiores nos dias 0, 3 e 7 do experimento. Nos grupos CT e DPT, as administrações tópicas foram realizadas com uma suspensão sham (sem probiótico). Todos os animais foram submetidos à eutanásia 14 dias após o início do experimento. Foram coletados tecido gengival, hemi-mandíbulas e biofilme bucal para avaliação dos seguintes parâmetros: i) microbiota bacteriana (checkerboard DNA-DNA hybridization; ii) expressão de citocinas pró e anti-inflamatórias (análise Multiplex); iii) imunorreatividade de peptídeos antimicrobianos (reações imunohistoquímicas - estreptavidina-biotina-peroxidase); iv) níveis inserção conjuntiva (análise histomorfométrica) e v) microarquitetura e volume ósseos (microtomografia computadorizada por transmissão de raios X). Os dados obtidos foram submetidos à análise estatística (p < 0,05). O grupo DPT apresentou maiores valores de porosidade óssea, espaçamento de trabéculas ósseas e nível de inserção conjuntiva, bem como menor número de trabéculas e volume ósseos quando comparado a todos os outros grupos (p < 0,05). No Grupo DPT-HN019, foram observados maiores percentuais de bactérias dos complexos amarelo e azul, bem como maiores expressões de Osteoprotegerina (OPG), Beta-defensina (BD)-1, BD-2 e BD-3 quando comparados àqueles do Grupo DPT (p < 0,05). O Grupo DPT apresentou níveis de Interleucina (IL)-1β Receptor Ativador do Fator Nuclear Kappa-beta (RANKL) maiores que aqueles do Grupo DPT-HN019 (p < 0,05). As razões RANKL/OPG e IL-1β/IL-10 foram mais elevadas no grupo DPT do que no Grupo DPT-HN019 (p < 0,05). Dentro dos limites deste estudo pode-se concluir que o uso tópico de B. lactis HN019 promove um efeito protetor contra a perda óssea e de inserção conjuntiva decorrentes da periodontite experimental em ratos.
The purpose of this study was to evaluate the effects of topical administration of probiotic bacteria of the genus Bifidobacterium on experimental periodontal disease in rats. 32 rats were divided into groups C (control), EP (experimental periodontitis), C-HN019 (control + probiotic) and EP-HN019 (EP+ probiotic). On day 0 of the experiment, periodontitis was induced in the animals of groups EP and EP-HN019 through the placement of ligatures around mandibular first molars. In groups C-HN019 and EP-HN019, 2 mL of suspensions containing 109 colony-forming units/mL of Bifidobacterium animalis subsp lactis (B. lactis) HN019 were topically administered in the subgingival region of mandibular first molars on days 0, 3 and 7 of the experiment. In groups C and EP, topical administrations were performed using a sham suspension (without probiotic). All animals were euthanized 14 days after the beginning of the experiment. Gingival tissue, hemi-mandibles and oral biofilm were collected for evaluation of the following parameters: i) bacterial microbiota (checkerboard DNA-DNA hybridization), ii) expression of pro- and anti-inflammatory cytokines (Multiplex analysis), iii) immunoreactivityof antimicrobial peptides (immunohistochemical reactions - streptavidin-biotin-peroxydase); iV) connective tissue attachment levels (histomorphometric analysis) and v) bone microarchitecture and volume (microtomographic analysis). Data were statistically analyzed (p < 0.05). Group EP presented greater values of bone porosity, trabecular separation and connective tissue attachment loss as well as reduced trabecular number and bone volume when compared with all the other groups (p < 0.05). In group EP-HN019, there were greater percentages of bacteria of the yellow and blue complexes and greater expressions of Osteoprotegerin (OPG) and beta-defensins (BD)-1, BD-2 and BD-3 when compared with group EP (p < 0.05). Group EP presented greater levels of Interleukin (IL)-1β and Receptor Activator of Nuclear Factor-Kappa B ligand (RANKL) than group EP-HN019 (p < 0.05). The increase in the ratios RANKL/OPG and IL-1β/IL-10 was greater in group EP than in group EP-HN019 (p < 0.05). Within the limits of the present study, it can be concluded that the topical use of B. lactis HN019 promotes a protective effect against alveolar bone and connective tissue attachment losses attributable to experimental periodontitis in rats.

The hypocholesterolemic effect of peptides present in crude casein hydrolysate using trypsin from Bifidobacterium animalis subsp. lactis (Bb-12) culture grown in casein solution or in MRS broth were compared with the control (unhydrolysed crude casein). These samples have shown to reduce cholesterol level in vitro to varying degree between 24-87%. The unhydrolysed crude casein (control) has shown not to have a reducing effect on cholesterol level. The reduction level of cholesterol was found to be dependant on the degree of hydrolysis. Fractions obtained after size exclusion from crude casein hydrolysate formed by trypsin after 48h hydrolysis have shown to reduce cholesterol level to degree vary between 2.7% to 50%. Bb-12 or trypsin crude casein hydrolysates were also found to possess angiotensin-converting enzyme (ACE) inhibitory activity in vitro to a varying degree between 29% and 38%, respectively. Unhydrolysed crude casein showed no real effect on ACE inhibitory activity. Trypsin hydrolysate has slightly more effect on ACE activity than Bb-l2. A relationship was also found between ACE-inhibitory activity and degree of hydrolysis. Fractions of crude casein hydrolysate after 48h hydrolysis with trypsin by size exclusion have shown to have ACE-inhibitory activity varying between 37% and 50%. Several identified ACE-inhibitory peptides from active fractions were found to have Phe, Trp, Tyr, Arg, Lys or Pro at their ultimate C-terminal position, making them a possible candidate for ACE-inhibitory activity. Further analysis on the profiles of casein and whey protein fractions, obtained by fast protein liquid chromatography (FPLC), of skimmed milk and four different commercial types of probiotic health drinks, and casein profile of added Bb-12 culture were investigated to study the possible effect of probiotics on milk proteins. Differences in protein-profile were found between the control and the four commercial probiotic health drinks as well as with added Bb-12 culture. Whey proteins were more resistance to hydrolysis than caseins.

Research Doctorate - Doctor of Philosophy (PhD)
Probiotics are increasingly being included into food products in order to develop functional foods with health promoting effects, but have to date been exploited mainly in dairy products. Development of non-dairy probiotic foods such as fruit juices may provide consumers with greater choice and be attractive to those who can not eat dairy foods. Orange juice presents as an ideal vehicle for probiotic delivery as it is the most popular fruit beverage worldwide, and like other fruit juices has a short gastro-intestinal transit time which reduces exposure of probiotics to harsh environment of the stomach. Since probiotic organisms vary in the type and level of their health promoting effects, it is likely that probiotic combinations would offer the consumer more benefit than single strains. Effective design of functional foods containing probiotic combinations, must take into consideration the likely occurrence and impact of potential interactions between individual species within a proposed combination, and between the probiotic and the carrier matrix. The main objectives of the current study were 1) to identify the effect of combining probiotics on their viability and adhesion to intestinal cells and 2) to examine the combined effect of exposure of probiotics to orange juice and low temperatures during refrigerated storage, on their viability and functional properties. The initial study of long-term (14 days) growth interactions of several lactobacilli and Bifidobacterium animalis subsp lactis Bb12 (Bb), both alone and in co-culture with Propionibacterium jensenii 702 (PJ), revealed that growth patterns of Lactobacillus strains were not adversely affected by the presence of PJ, whereas lactobacilli strongly inhibited growth of PJ. In the co-culture of Bb and PJ, a significant enhancement of the growth of both bacteria was observed. The effect of combining probiotics on their adhesion to human intestinal epithelial Caco-2 cells was only evident in the case of Lb. casei 01 and Lb. rhamnosus GG (LG) which exhibited a decrease in adhesion rate in the presence of PJ. The viability of LG, Lb. reuteri ATCC 55730 (LR), Bb and PJ, both individually and as 2- or 3- multispecies combinations, were then monitored in orange juice (OJ) (with and without 20% pulp) as well as bottled drinking water (BW) over 8 weeks of refrigerated (4ºC) and non-refrigerated storage (only for BW). Lactobacilli remained viable in higher numbers in OJ relative to that observed in BW under refrigeration. In contrast, a better outcome was observed for Bb and PJ in BW. Combining of probiotic species was observed to affect individual strain viability. Presence of pulp did not affect the viability of probiotics in OJ, while storage of BW at room temperature had an adverse effect on viability of all probiotics except of PJ, relative to storage under refrigeration. Influence of combined exposure to OJ and refrigerated storage of the same probiotic preparations on their in vitro gasto-intestinal tolerance, adhesion to intestinal epithelial cells and immunomodulatory effects was then investigated at 10-day intervals during one month of storage. Suspension in OJ did not adversely affect the tolerance of any of the strains examined to simulated gastric juice (SGJ), with the tolerance of LG and PJ considerably enhanced relative to that observed in PBS, but did appear to impair the tolerance of lactobacilli and PJ to simulated intestinal juice (SIJ) at the baseline. High tolerance to SGJ was maintained throughout the storage period. The tolerance of both Bb and PJ to SIJ remained relatively constant during storage. Combining with both Bb and PJ enhanced the tolerance of the lactobacilli to SIJ with little impact on Bb, but adversely affected PJ in all combinations. The adhesion rate of LG remained relatively constant in all preparations along with the viability during storage. In contrast with LG, adhesion rates and viabilities of other probiotics exhibited variation in relation to strain, presence of other microorganisms, and storage duration. In terms of both viability and adhesion rate, the preparations that provided the best outcomes for all constituents were LG and LR-PJ. With the exception of LG, all probiotic preparations significantly enhanced non-stimulated interleukin-8 (IL-8) but not interleukin-6 (IL-6) or tumor necrosis factor-α (TNF-α) secretion by Caco-2 cells. Probiotic preparations enhanced Escherichia coli lipopolysaccharide (LPS) induced IL-8 release at baseline however this effect was not evident in all preparations at day 10. With the exception of LG, all probiotic preparations enhanced TNF-α induced IL-8 secretion towards day 20 after which it returned to the control level. In contrast, probiotic preparations significantly reduced IL-1β induced IL-8 secretion at baseline, with no further effect evident during storage. The relative probiotic effect on IL-1β and TNF-α induced IL-8 secretion showed an upward and downward trend respectively over the storage period. Probiotic preparations did not affect LPS or IL-1β induced secretion of IL-6 up to 10 days of storage, while thereafter some of them exhibited variable effects on IL-1β induced IL-6 secretion. Compared to baseline (day 0), the effect of all four probiotic strains on IL-1β induced TNF-α production was found to decrease significantly by day10 of the storage period. In conclusion, the results provided evidence of variation between individual strains in terms of their viability and intestinal adhesion capacity, and for the same strain when combined with different probiotics. When included in bottled drinking water and orange juice, the viabilities and functional properties of the probiotic preparations were further affected by the duration of their exposure to the carrier matrix and refrigerated storage. Such effects should be considered when formulating probiotic products, and further research is recommended to confirm the observed in vitro functional effects in vivo.