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

No description available.

In recent years, there has been an upsurge in medical research assessing the therapeutic benefits of probiotic bacteria and growing commercial interest in food fortification with these bacteria. Probiotic bacteria such as L. acidophilus are known to be predominant Lactobacilli species in the intestinal tract of healthy humans and suggested to provide clinical health benefits such as enhancement of immunity against intestinal infections, prevention of diarrhea and hypercholesterolaemia and improvement in lactose utilization. Many studies have demonstrated the possibility of incorporating probiotic bacteria in an ice cream matrix and shown its viability can be maintained throughout the shelf life of the ice cream. However, there is limited information about the protective effect of ice cream on viability of incorporated probiotic bacteria during simulated gastric digestion using an in vitro dynamic model stomach. In phase one of this study, a preliminary study was conducted to determine the effect of air addition on the viability of L. acidophilus La-5. This was done by manufacturing low fat (4%) non-fermented ice cream mix supplemented with L. acidophilus La-5 to yield an initial population of 107cfu/g. The mix was processed with 60% and 100% overrun (OR) and stored at -10ᵒC for 90 days. The effect of air addition at different levels was tested post freezing and every 30 days throughout its shelf life of 90 days. The results showed less than one log reduction in the viable counts of L. acidophilus La-5 for both samples incorporated with 60% and 100% OR after freezing and the number of viable cells did not differ significantly (p>0.05) from day 1 to day 90. In phase two of this study, a 22 full factorial experimental design was used to evaluate whether the viscous nature of ice cream mix plays an important role in improving the survivability of L. acidophilus La-5 during simulated digestion against low pH and presence of mechanical shear and to determine whether initial inoculation level has any effect on the viability of L. acidophilus La-5 at the end of 2 hr simulated digestion. Non-fermented low fat (5%) ice cream mixes with high and low viscosity were produced by changing the amount of stabilizer/emulsifier blend and each of the two mixes were supplemented with two levels of L. acidophilus La-5 to obtain an initial population of 108cfu/g and 106cfu/g before freezing. These mixes were frozen with 60% overrun. The ice cream samples were digested for 2 hr in an in vitro model stomach called Human Gastric Simulator (HGS). This model included factors such as gastric secretions, mechanical shearing due to peristaltic contractions and temperature and pH control. No significant effect (p>0.05) of different levels of viscosity on the survivability of L. acidophilus La-5 was found during and at the end of 2 hr simulated in vitro digestion, irrespective of the difference in initial inoculation level. The initial supplementation level of L. acidophilus La-5 had a significant impact (p<0.05) on its survivability during the simulated digestion of ice cream samples, irrespective of the difference in viscosity. The log survival of L. acidophilus La-5 was on an average 3.64 log cfu/g and 4.08 log cfu/g for ice cream samples supplemented with higher and lower amount of L. acidophilus La-5, respectively at the end of 2 hr. Nevertheless, this difference in overall survival was not statistically significant (p>0.05). These studies demonstrated the efficacy of low fat non-fermented ice cream in maintaining high viable numbers of L. acidophilus La-5 throughout its tested shelf life of 90 days. In addition, protective effect of ice cream on the viability of L. acidophilus La-5 against harsh stomach conditions was observed, but this effect was not as a result of viscosity of ice cream. It was also found that an ice cream supplemented with 106cfu/g would result in a similar overall log reduction of L. acidophilus La-5 at the end of 2 hr simulated digestion compared to an ice cream supplemented with 108cfu/g. The aggressive stomach conditions had a negative impact on the survivability of L. acidophilus La-5 during digestion of all the ice cream samples, but this detrimental effect can be reduced by incorporating L. acidophilus La-5 into an ice cream matrix which would increase the opportunity of bacteria to reach the small intestine and provide the desired health benefit.

L’usage des produits microbiens administrés directement (aussi appelés probiotiques) gagne de l’intérêt comme alternative à l’utilisation des antibiotiques comme promoteurs de croissance dans les élevages. Cependant, très peu d’informations existent quant à l’influence des probiotiques sur la modulation du microbiote gastrointestinal et la réponse immunitaire innée chez le veau laitier. Les objectifs de cette thèse visaient à (1) Étudier l’effet de Lactobacillus acidophilus BT 1386 ou de Saccharomyces cerevisiae boulardii CNCM 1- 1079 sur les constituants sanguins, biochimiques / chimiques du sang. (2) Déterminer les mécanismes potentiels d’une réponse immunitaire renforcée de Lactobacillus acidophilus BT 1386 et de Saccharomyces cerevisiae boulardii CNCM 1-1079. (3) Déterminer comment Lactobacillus acidophilus BT 1386 ou Saccharomyces cerevisiae boulardii CNCM 1-1079 modulent la composition de la communauté microbienne GIT de veau par séquençage de nouvelle génération de la région V3-V4 du gène ARNr 16S. (4) Comparer l'efficacité de ces deux DFM avec la tetracycline-néomycine, un promoteur de croissance antibiotique. Quatre traitements ont été distribués aléatoirement à 48 veaux âgés de 2 à 7 jours (n=12). TÉMOIN : lactoremplaceur (LR) suivi d’une moulée de démarrage (MD); SCB) TÉMOIN + Saccharomyces cereviseae var. boulardii CNCM I-1079 [7,5 × 108 unités formatrices de colonie (CFU)/L de LR + 3 × 109 CFU/kg de MD]; LA) TÉMOIN + Lactobacillus acidophilus BT 1386 (2,5 × 108 CFU/L de LR + 1 × 109 CFU/kg de MD); ATB) TÉMOIN + traitement antibiotique composé de chlortétracycline (528 mg/L de LR + 55 mg/kg de MD) et de néomycine (357 mg/L de LR). Les animaux ont été élevés selon les procédures d’élevage conventionnelles pendant les 96 jours de la période expérimentale. Des échantillons de sang ont été prélevés de la veine jugulaire à différents moments pendant les périodes de pré-sevrage (jours 1 à 42), de sevrage (jours 43 à 53) et de post-sevrage (jours 54 à 96). Aux jours 33 et 96 dans chacun des groupes, 4 veaux ont été euthanasiés afin de prélever des échantillons de tissus et de digesta. Des SCB viables ont été retrouvées tout au long du tractus gastrointestinal, ainsi que dans les fèces des veaux en périodes pré- et post-sevrage. Autour du sevrage, les fèces du groupe SCB contenaient une population de lactobacilli plus importante que celles du groupe TÉMOIN. Au cours de la période pré-sevrage, la distribution des lactobacilli évoluait graduellement à travers les sections du tube digestif (colon > contenu iléal > rumen > muqueuse iléale). À l’exception du rumen, tous les autres compartiments présentaient une population de lactobacilli réduite en post- vs. en pré-sevrage. Comparativement aux groupes TÉMOIN et LA, la profondeur et la largeur des cryptes du colon des groupes SCB et ATB étaient réduites. Toujours comparativement aux groupes TÉMOIN et LA, le nombre de cellules caliciformes contenant des mucines neutres tendait à augmenter pour les groupes SCB et ATB, alors que le nombre de mucines acides augmentaient. Globalement, les traitements n’ont pas affecté les performances des animaux. Pendant le sevrage, une amélioration de la stimulation oxydative et de la phagocytose, ainsi qu’une augmentation des concentrations des protéines de la phase aiguë, ont été observées chez les groupes SCB et LA. L’ajout de probiotiques à la diète du veau a eu moins d’impact sur la diversité bactérienne mais a tout de même modifié significativement l’abondance des différentes populations microbiennes, et ce plus particulièrement dans l’iléon. L’ajout de SCB ou de LA a réduit l’abondance de certains genres bactériens pathogènes, tels que Streptococcus et Tyzzerella_4, alors que cela a augmenté l’abondance de bactéries potentiellement bénéfiques pour l’hôte tel que celles appartenant au genre Fibrobacter. Par ailleurs, d’autres bactéries bénéfiques tel que Rumminococcaceae UCG 005 et Olsenella étaient aussi plus abondantes, mais seulement pour le traitement SCB. Les bactéries pathogènes Peptoclostridium et Ruminococcus_2 étaient respectivement moins abondantes lorsque les traitements SCB et LA étaient ajoutés à la ration. Les analyses de prédiction fonctionnelle ont montré qu’en plus des effets observés sur les voies métaboliques locales impliquées dans le cycle cellulaire, la sécrétion biliaire et les voies de signalisation de l’AMPc et du proteasome, l’ajout des deux formes de probiotiques a également affecté d’importantes voies impliquées au sein d’autres tissus comme la synthèse des hormones thyroïdiennes ou le fonctionnement des synapses dopaminergiques. Cette étude suggère que les probiotiques, et plus particulièrement SCB, devraient être davantage considérés comme modulateur de la santé gastro-intestinale du veau laitier. Aussi, la supplémentation en SCB, en améliorant la réponse immunitaire innée, permettrait de stimuler le système immunitaire du veau avant l’infection, le préparant ainsi à mieux affronter les périodes plus sensibles comme celle du sevrage. Le SCB et le LA ont modifié la composition en bactéries du GIT. Dans l’ensemble, cette étude a montré une démonstration remarquable de l’importance du DFM sur le microbiote de la TI. Cependant, il faut mieux comprendre les molécules et les mécanismes qui déterminent le rôle du microbiote, puis exploiter ces connaissances pour améliorer la santé et augmenter la production animale / L’usage des produits microbiens administrés directement (aussi appelés probiotiques) gagne de l’intérêt comme alternative à l’utilisation des antibiotiques comme promoteurs de croissance dans les élevages. Cependant, très peu d’informations existent quant à l’influence des probiotiques sur la modulation du microbiote gastrointestinal et la réponse immunitaire innée chez le veau laitier. Les objectifs de cette thèse visaient à (1) Étudier l’effet de Lactobacillus acidophilus BT 1386 ou de Saccharomyces cerevisiae boulardii CNCM 1- 1079 sur les constituants sanguins, biochimiques / chimiques du sang. (2) Déterminer les mécanismes potentiels d’une réponse immunitaire renforcée de Lactobacillus acidophilus BT 1386 et de Saccharomyces cerevisiae boulardii CNCM 1-1079. (3) Déterminer comment Lactobacillus acidophilus BT 1386 ou Saccharomyces cerevisiae boulardii CNCM 1-1079 modulent la composition de la communauté microbienne GIT de veau par séquençage de nouvelle génération de la région V3-V4 du gène ARNr 16S. (4) Comparer l'efficacité de ces deux DFM avec la tetracycline-néomycine, un promoteur de croissance antibiotique. Quatre traitements ont été distribués aléatoirement à 48 veaux âgés de 2 à 7 jours (n=12). TÉMOIN : lactoremplaceur (LR) suivi d’une moulée de démarrage (MD); SCB) TÉMOIN + Saccharomyces cereviseae var. boulardii CNCM I-1079 [7,5 × 108 unités formatrices de colonie (CFU)/L de LR + 3 × 109 CFU/kg de MD]; LA) TÉMOIN + Lactobacillus acidophilus BT 1386 (2,5 × 108 CFU/L de LR + 1 × 109 CFU/kg de MD); ATB) TÉMOIN + traitement antibiotique composé de chlortétracycline (528 mg/L de LR + 55 mg/kg de MD) et de néomycine (357 mg/L de LR). Les animaux ont été élevés selon les procédures d’élevage conventionnelles pendant les 96 jours de la période expérimentale. Des échantillons de sang ont été prélevés de la veine jugulaire à différents moments pendant les périodes de pré-sevrage (jours 1 à 42), de sevrage (jours 43 à 53) et de post-sevrage (jours 54 à 96). Aux jours 33 et 96 dans chacun des groupes, 4 veaux ont été euthanasiés afin de prélever des échantillons de tissus et de digesta. Des SCB viables ont été retrouvées tout au long du tractus gastrointestinal, ainsi que dans les fèces des veaux en périodes pré- et post-sevrage. Autour du sevrage, les fèces du groupe SCB contenaient une population de lactobacilli plus importante que celles du groupe TÉMOIN. Au cours de la période pré-sevrage, la distribution des lactobacilli évoluait graduellement à travers les sections du tube digestif (colon > contenu iléal > rumen > muqueuse iléale). À l’exception du rumen, tous les autres compartiments présentaient une population de lactobacilli réduite en post- vs. en pré-sevrage. Comparativement aux groupes TÉMOIN et LA, la profondeur et la largeur des cryptes du colon des groupes SCB et ATB étaient réduites. Toujours comparativement aux groupes TÉMOIN et LA, le nombre de cellules caliciformes contenant des mucines neutres tendait à augmenter pour les groupes SCB et ATB, alors que le nombre de mucines acides augmentaient. Globalement, les traitements n’ont pas affecté les performances des animaux. Pendant le sevrage, une amélioration de la stimulation oxydative et de la phagocytose, ainsi qu’une augmentation des concentrations des protéines de la phase aiguë, ont été observées chez les groupes SCB et LA. L’ajout de probiotiques à la diète du veau a eu moins d’impact sur la diversité bactérienne mais a tout de même modifié significativement l’abondance des différentes populations microbiennes, et ce plus particulièrement dans l’iléon. L’ajout de SCB ou de LA a réduit l’abondance de certains genres bactériens pathogènes, tels que Streptococcus et Tyzzerella_4, alors que cela a augmenté l’abondance de bactéries potentiellement bénéfiques pour l’hôte tel que celles appartenant au genre Fibrobacter. Par ailleurs, d’autres bactéries bénéfiques tel que Rumminococcaceae UCG 005 et Olsenella étaient aussi plus abondantes, mais seulement pour le traitement SCB. Les bactéries pathogènes Peptoclostridium et Ruminococcus_2 étaient respectivement moins abondantes lorsque les traitements SCB et LA étaient ajoutés à la ration. Les analyses de prédiction fonctionnelle ont montré qu’en plus des effets observés sur les voies métaboliques locales impliquées dans le cycle cellulaire, la sécrétion biliaire et les voies de signalisation de l’AMPc et du proteasome, l’ajout des deux formes de probiotiques a également affecté d’importantes voies impliquées au sein d’autres tissus comme la synthèse des hormones thyroïdiennes ou le fonctionnement des synapses dopaminergiques. Cette étude suggère que les probiotiques, et plus particulièrement SCB, devraient être davantage considérés comme modulateur de la santé gastro-intestinale du veau laitier. Aussi, la supplémentation en SCB, en améliorant la réponse immunitaire innée, permettrait de stimuler le système immunitaire du veau avant l’infection, le préparant ainsi à mieux affronter les périodes plus sensibles comme celle du sevrage. Le SCB et le LA ont modifié la composition en bactéries du GIT. Dans l’ensemble, cette étude a montré une démonstration remarquable de l’importance du DFM sur le microbiote de la TI. Cependant, il faut mieux comprendre les molécules et les mécanismes qui déterminent le rôle du microbiote, puis exploiter ces connaissances pour améliorer la santé et augmenter la production animale. / There is interest in the use of direct-fed microbials (DFM) as substitutes for antibiotic growth promoters in farm animal production. However, little information exists on the effects of Lactobacillus acidophilus BT 1386 (LA) and Saccharomyces cereviseae boulardii CNCM I-1079 (SCB) on the modulation of the gastrointestinal tract (GIT) microbiota and innate immune responses in dairy calves. Therefore, the objectives of this thesis were to (1) investigate the effect of Lactobacillus acidophilus BT 1386 or Saccharomyces cerevisiae boulardii CNCM 1-1079 on blood cellular and biochemical/chemical constituents; (2) determine the potential mechanisms of enhanced immune response by Lactobacillus acidophilus BT 1386 and Saccharomyces cerevisiae boulardii CNCM 1-1079; (3) determine how Lactobacillus acidophilus BT 1386 or Saccharomyces cerevisiae boulardii CNCM 1-1079 modulate calf GIT microbial community composition by next-generation sequencing of the V3-V4 region of the 16S rRNA gene and (4) compare the efficacy of these two DFM with tetracycline-neomycin, an antibiotic growth promoter. Forty eight calves (2 to 7 days old) were randomly allocated to four treatments: 1) Control (CTRL) fed milk replacer (MR) and starter feed (SF); 2) CTRL supplemented with Saccharomyces cerevisiae boulardii CNCMI-1079 (SCB; 7.5 × 108 (CFU)/L MR + 3 × 109 CFU/kg SF); 3) CTRL supplemented with Lactobacillus acidophilus BT1386 (LA; 2.5 × 108 CFU/L MR + 1 × 109 CFU/kg SF); and 4) CTRL supplemented with antibiotics (ATB) chlortetracycline and neomycin (528 and 357 mg/L MR, respectively), and chlortetracycline (55 mg/kg SF). Animals were raised for 96 days following standard management procedures. Growth parameters (body weight and feed intake) of calves were recorded weekly. Four calves per treatment were euthanized on day 33 (pre-weaning) and an additional four calves per treatment on day 96 (post-weaning) to sample rumen and ileum tissues for real time quantitative polymerase chain reaction and colon for histomorphology. The ileum, colon and rumen were also analyzed for viability. Furthermore, samples of digesta (colon, ileum and rumen) and mucosa (colon and ileum) for bacterial characterization by sequencing the V3-V4 region of 16S rRNA gene. Weekly feces samples were collected for viability analysis. Blood samples were also collected for isolation of neutrophils and peripheral blood mononuclear cells for oxidative burst and phagocytosis analyses by flow cytometry. Serum measurements of acute phase proteins were done by ELISA. Viable SCB were recovered throughout the GIT and in the feces pre- and post-weaning. The feces of SCB-treated calves showed a greater lactobacilli population compared with CTRL (P < 0.01) around weaning. In the pre-weaning period, the distribution of lactobacilli population differed along the digestive tract (colon > ileum content > rumen > ileum mucosa; P < 0.001). The lactobacilli population were significantly reduced in all compartments (P = 0.02) post-weaning compared to pre-weaning, except in the rumen. Crypts depth and width of the colon decreased (P < 0.01) whereas number of goblet cells containing neutral mucins tended to increase (P = 0.058) while acidic mucins increased (P < 0.05) in SCB- and ATB-treated calves compared with CTRL and v LA-treated calves. Overall, growth performances were not affected by treatment. There was improvement of both oxidative burst and phagocytosis by SCB and LA during weaning in calves. Similarly, the concentrations of acute phase proteins (C-reactive proteins and serum amyloid A proteins) were increased by SCB and LA during weaning. The DFM had less impact on the bacteria diversity but had significant impact on the abundance of the bacteria community with most changes associated to treatments occurring in the ileum. SCB and LA reduced some pathogenic bacteria genera such as Streptococcus, Tyzzerella_4 and increased some potential beneficial bacteria such as fibrobacter. Meanwhile, Rumminococcaceae UCG 005 and Olsenella, also beneficial, were increased only by SCB treatment. The potential pathogenic bacterium, Peptoclostridium, was reduced by SCB only while LA reduced Ruminococcus_2. The functional prediction analyses indicated that besides affecting local pathways such as cell cycle, bile secretion, proteasome or cAMP signaling pathway, both DFM might also affect important pathways in other tissues such as thyroid hormone synthesis or Dopaminergic synapse in the brain. Our results suggest that SCB is a modulator of gastrointestinal health and could prime the immune system prior to infection leading to an enhanced innate immune response in calves especially during periods of stress (e.g., weaning). Consequently, SCB might have the potential to strengthen calf immune system in the critical periods of disease susceptibility. Both SCB and LA changed the bacteria composition of the GIT. Overall, this study showed a remarkable demonstration of the importance of DFM on the GIT microbiota. However, what is needed is a complete and better understanding of the molecules and mechanisms driving the roles played by the microbiota and then to exploit this knowledge to improve health and increase animal production.

The causality dilemma between dysbiosis and cancer has given rise to numerous studies both exploring the mechanisms behind cancer progression and the associative shifts in the microbiota upon carcinogenesis. Aside from the hallmark study of Dr. Barry Marshall in establishing the true causal relationship between Helicobacter pylori and gastric adenocarcinoma, studies have only been successful in adding associative links of carcinogenesis mediated by bacteria to the literature. The current field is limited in its ability to establish causative relationships, and further work is needed to construct a reference community whose physiological responses reflect global community responses. In this thesis, the organism Lactobacillus acidophilus was selected as a pilot strain for the development of a novel framework to establish the fitness and physiological changes that occur when bacteria engage the human epithelial environment. The pilot strain was revived from the American Type Culture Collection (ATCC), verified through 16S rRNA Sanger sequencing, and grown in its conventional culture medium and human tissue culture medium to establish baseline growth rates and gauge its physiological responses to an in vitro tumor microenvironment. A set of standard conditions was proposed for growth under human tissue culture conditions. Finally, a metabolic study and spot plate assay were performed to elucidate the anabolic deficits and viability of this strain in human tissue culture medium, respectively. This research was performed to better understand the environmental and metabolic requirements for this pilot strain to inhabit the human epithelial environment, and to establish a workflow that will set the foundation for an appropriate clinical study to demonstrate the causative relationship between dysbiosis and carcinogenesis.

Le lactosérum est un coproduit de l'industrie de fabrication du fromage et de la caséine et se caractérise par une forte demande chimique et biologique en oxygène. Les énormes quantités de lactosérum générées dans le monde, sa composition particulière et son utilisation limitée dans l'industrie alimentaire rendent nécessaire la recherche d'autres moyens d'ajouter de la valeur à cet ingrédient en vue d'augmenter la rentabilité de la transformation du lait. Dans ce contexte, la technologie de l'électro-activation (EA) offre la possibilité de valoriser le lactosérum par la conversion in situ d'une partie du lactose en lactulose, un prébiotique bien connu et éprouvé. De plus, l'EA cathodique du lactosérum a montré une formation des bases de Schiff suite à la glycation avec différents sucres des protéines, des peptides et des acides aminés libres dans le processus d'électro-isomérisation du lactose en lactulose. Ces produits sont connus pour leur forte activité antioxydante. Ainsi, l'EA ouvre une possibilité de générer un ingrédient fonctionnel avec une valeur ajoutée significative. Dans ce contexte, l'objectif principal de ce projet de doctorat était d'étudier et de démontrer la faisabilité technologique de l'utilisation du lactosérum électro-activé comme ingrédient fonctionnel à haut potentiel prébiotique dans la production de différents produits laitiers fermentés. La première étape de ce projet a été l'évaluation du comportement du lactosérum électro-activé dans la matrice de gel de lait fermenté. Une comparaison entre le pourcentage de matière grasse du lait, l'inoculum de lactosérum et le type de lactosérum a été effectuée. À cette fin, des échantillons de lait fermenté ont été préparés avec un ajout de 3, 6 et 9 % de lactosérum des deux types (électro-activé et non électro-activé). Il a été constaté que le lactosérum électro-activé prolongeait le temps d'obtention d'un pH de 4,6 en fonction de la quantité ajoutée. Ceci a été attribué à la capacité tampon plus élevée du lactosérum électroactivé; les résultats de l'acidité titrable ayant démontré des niveaux élevés de groupes acides libres. La microstructure du gel obtenu avec l'ajout du lactosérum électro-activé a montré une structure uniforme et moins poreuse, ce qui était en accord avec les résultats de la réduction de la synérèse. Pour confirmer ces résultats, un autre produit laitier fermenté avec un ajout de lactosérum électro-activé a été également développé. Le kéfir enrichi de lactosérum électro-activé présentait également une phase de fermentation prolongée. Les particules de lactosérum EA ont été incorporées de manière homogène dans la matrice du gel de kéfir. Par conséquent, aucune synérèse n'était visible dans les échantillons de kéfir additionnés de lactosérum EA à 9 %. De plus, les deux produits contenaient des niveaux élevés d'acides organiques (lactique, citrique, acétique, propionique et butyrique) lorsqu'ils étaient supplémentés avec du lactosérum EA. La production d'acide butyrique a été induite par l'ajout de lactosérum des deux types. L'analyse HPLC a révélé qu'environ 75-85% des niveaux initiaux de lactulose ont été conservés dans les produits avec du lactosérum EA après le processus de fermentation, ce qui démontre que la consommation de tels produits pourrait constituer une source de lactulose pour le consommateur. La deuxième étape de cette recherche a été d'optimiser l'utilisation du lactosérum électro-activé en tant qu'ingrédient par son incorporation dans le produit qui convient à sa couleur et aux caractéristiques de la réaction de Maillard et des conjugués entre les matières azotées avec les sucres. Le lait fermenté cuit, Ryazhenka, a été testé comme une matrice alimentaire appropriée pour véhiculer le lactosérum EA enrichi en lactulose. L'extension du temps de fermentation a été moins importante pour ce produit. Ainsi, le Ryazhenka additionné de lactosérum à 9% a atteint un pH de 4,6 après 4 h de fermentation. Le produit additionné de lactosérum EA (9%) a atteint ce niveau après 6,5 h. De plus, le lactosérum EA a amélioré la capacité antioxydante de Ryazhenka. Au cours de cette étape, nous avons démontré par des tests in vitro que l'électro-activation du lactosérum peut diminuer l'allergénicité de la β-lactoglobuline de 19,52 mg/kg à 7,56 mg/kg, qui s'est stabilisée à 12,13 mg/kg après neutralisation. Comme le protocole de production de Ryazhenka comprend une étape de cuisson de 3 à 5 h à 97-100°C, on considère qu'il présente des taux d'allergénicité plus faibles en raison des changements de conformation des protéines induits par la chaleur. Ainsi, l'ajout de lactosérum électro-activé ne contribue pas à l'augmentation de l'allergénicité de ce produit. Le troisième objectif de cette étude était de démontrer un potentiel prébiotique du lactosérum électro-activé en cultivant des bactéries probiotiques Lactobacillus rhamnosus subsp, Lactobacillus rhamnosus GG et Lactobacillus acidophilus ATCC4356. La densité optique(OD₆₀₀), le dénombrement sur plaques de Petri, la stabilité durant l'entreposage à 4 °C et la tolérance aux acides et à la bile des bactéries cultivées pendant 24 heures dans du lactosérum électro-activé ont été étudiés et comparés aux résultats obtenus par la culture sur du lactosérum, du lactosérum additionné de lactulose, du MRS et du MRS avec ajout de lactulose. Les valeurs OD₆₀₀ les plus élevées (>2) ont été obtenues dans les biomasses de lactosérum EA pour toutes ces bactéries. Cependant, les numérations sur plaque de Petri n'ont pas confirmé un nombre plus élevé de cellules bactériennes dans du lactosérum électro-activé. On peut donc conclure que le lactosérum électro-activé a probablement stimulé un métabolisme distinct chez les bactéries testées, ce qui est conforme à la définition des prébiotiques qui ont la particularité d'induire une stimulation de la croissance et/ou de l'activité des bactéries probiotiques afin de conférer des avantages pour la santé. En résumé, cette recherche a validé la faisabilité technologique de l'utilisation du lactosérum électro-activé comme ingrédient dans la production de lait fermenté et source de lactulose qui reste stable durant l'entreposage pendant 14 jours à 4 °C. Également, ce projet a montré que le lactosérum électro-activé est un ingrédient fonctionnel prometteur pour une éventuelle utilisation potentielle comme additif alimentaire fonctionnel et prébiotique dans l'industrie laitière. De plus, il peut être utilisé comme agent protecteur pour améliorer la viabilité et l'activité des probiotiques.

O objetivo do presente estudo clínico randomizado foi avaliar in vivo, por meio da técnica de biologia molecular Checkerboard DNA-DNA Hybridization: 1) A contaminação de bráquetes metálicos por 4 espécies bacterianas de microrganismos cariogênicos (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei e Lactobacillus acidophilus); e 2) A eficácia da utilização do gluconato de clorexidina a 0,12% (Periogard&reg;) sob a forma de bochechos, sobre esses microrganismos. Participaram do estudo 39 pacientes de 11 a 33 anos de idade, em tratamento com aparelho ortodôntico fixo, nos quais foram colados 2 bráquetes metálicos novos, nos pré-molares. Os pacientes do Grupo Controle (n=20) foram orientados a fazer 2 bochechos semanais com solução placebo, durante 30 dias. Os pacientes do Grupo Experimental (n=19) foram orientados a fazer bochechos com solução à base de gluconato de clorexidina a 0,12% (Periogard&reg;), da mesma forma que o grupo Controle. Decorridos 30 dias, os 2 bráquetes foram removidos de cada paciente e processados para detecção das 4 cepas bacterianas, pela técnica Checkerboard DNADNA Hybridization. Os resultados obtidos foram analisados por meio do teste nãoparamétrico de Kruskal-Wallis, utilizando o software SAS (Statistical Analysis System). O nível de significância adotado foi de 5%. De acordo com os resultados obtidos, observou-se que S. mutans, S. sobrinus, L. casei e L. acidophilus foram detectados em 100% das amostras dos bráquetes de ambos os grupos. No entanto, os bráquetes do grupo Controle encontravam-se mais densamente contaminados por S. mutans e S. sobrinus (p<0,01). No grupo Experimental, embora as quantidades de S. mutans, S. sobrinus, L. casei e L. acidophilus tenham sofrido redução numérica após o uso dos bochechos com solução de gluconato de clorexidina a 0,12%, a comparação com os valores observados no grupo Controle evidenciou diferença estatisticamente significante apenas para S. mutans (p=0,03). Concluiu-se que os bochechos com solução de gluconato de clorexidina a 0,12% podem ser úteis, na prática clínica, com a finalidade de reduzir os níveis de microrganismos cariogênicos, em pacientes portadores de aparelhos ortodônticos fixos. / The purpose of this randomized clinical study was to evaluate in vivo, by the checkerboard DNA-DNA hybridization biomolecular technique: 1) The contamination of metallic brackets by four cariogenic bacterial strains (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei and Lactobacillus acidophilus); and 2) The efficacy of 0.12% chlorhexidine gluconate mouthrinses (Periogard&reg;) against these microorganisms. Thirty-nine 11-33-year-old patients under treatment with fixed orthodontic appliances were enrolled in the study and had 2 new metallic brackets bonded to the premolars. For the patients of the control group (n=20), mouthrinses with a placebo solution were prescribed twice a week during 30 days. The patients randomized to the experimental group (n=19) were instructed to use a 0.12% chlorhexidine gluconate solution (Periogard&reg;) as an oral rinse twice a week during 30 days, in the same way as in control group. After this period, the 2 brackets were removed from each patient and processed for detection of the 4 bacterial strains by checkerboard DNA-DNA hybridization. The obtained data were analyzed statistically by the non-parametric Kruskal-Wallis test using the SAS (Statistical Analysis System) software. A level of significance of 5% was set for all analysis. S. mutans, S. sobrinus, L. casei and L. acidophilus were detected in 100% of the samples from brackets of both groups. However, the brackets of the control group were more heavily contaminated by S. mutans and S. sobrinus (p<0.01). In the experimental group, although S. mutans, S. sobrinus, L. casei and L. acidophilus counts decreased after rinsing with the 0.12% chlorhexidine gluconate solution, the comparison with the values obtained in the control group showed statistically significant difference only for S. mutans (p=0.03). In conclusion, the use of 0.12% chlorhexidine gluconate mouthrinses can be useful in clinical practice to reduce the levels of cariogenic microorganisms in patients under treatment with fixed orthodontic appliances.

A possibilidade de se obter um produto com grande aceitabilidade nacional, principalmente voltado ao público infantil, com propriedades microbiológicas, nutricionais, físico-químicas e organolépticas ótimas associadas aos benefícios creditados aos probióticos é bastante promissora. Assim, o presente trabalho teve como objetivo verificar a influência do emprego de culturas probióticas compostas pelos microrganismos Lactobacillus acidophilus La-5 e Bifidobacterium animalis subsp. lactis BL04 isolados e em co-cultura, em queijo tipo petit-suisse processado com a adição de Streptococcus thermophilus como cultura starter, sobre as características do produto, ao longo do seu armazenamento refrigerado. Quatro tratamentos de queijo petit-suisse foram estudados (em triplicata): T1 (controle - Streptococcus thermophilus ST), T2 (ST + Lactobacillus acidophilus La-5), T3 (ST + B. animalis subsp. lactis BL04) e T4 (ST + La-5 + BL04) e armazenados a 4±1ºC. Foram avaliados parâmetros microbiológicos (população de probióticos, starter e contaminantes), de textura, físico-químicos e sensoriais (aceitabilidade). As análises sensoriais foram realizadas após 7 e 14 dias e as demais análises, após 1, 7, 14, 21 e 28 dias de armazenamento dos produtos a 4±1oC. As populações dos probióticos estiveram sempre superiores ao mínimo recomendado para um alimento probiótico, tendo variado de 7,22 a 7,60 log UFC/g e de 8,56 a 8,72 log UFC/g durante o armazenamento, para L. acidophilus e B. animalis subsp. lactis, respectivamente. A cultura starter apresentou populações entre 9,20 e 9,61 log UFC/g no mesmo período. As populações máximas para coliformes totais, bolores e leveduras foram de 2,79 e 3,60 log UFC/g, respectivamente. Os valores de pH diminuíram ao longo do armazenamento, devido à atividade dos microrganismos acidificantes presentes. A umidade variou entre 67,16% e 70,26% no mesmo período. Todos os queijos apresentaram queda na dureza e adesividade ao longo do armazenamento. Na análise sensorial, o queijo T4, com a co-cultura, revelou uma aceitabilidade significativamente maior em todos os atributos avaliados (p<0,05). Após 14 dias de armazenamento, o queijo T3 apresentou uma aceitabilidade significativamente inferior aos demais (p<0,05). A suplementação de queijo petit-suisse T4 com Lactobacillus acidophilus La-5 e B. animalis subsp. lactis BL04 em co-cultura com a cultura starter resultou em um produto com excelente potencial comercial, tendo apresentado características sensoriais, microbiológicas e de textura bastante vantajosas. / The possibility of obtaining a product with wide acceptability in Brazil, and which is mainly targeted at children and has excellent microbiological, nutritional, physical-chemical and organoleptic properties associated with the benefits ascribed to probiotic microorganisms is rather promising. Thus, the aim of the present work was to examine the influence of the probiotic cultures of Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BL04, isolated and in co-culture, on the characteristics of petit-suisse cheese produced with the addition of Streptococcus thermophilus as a starter culture, throughout the period of refrigerated storage. Four petit-suisse cheese trials were studied (in triplicates): T1 (control - Streptococcus thermophilus ST), T2 (ST + Lactobacillus acidophilus La-5), T3 (ST + Bifidobacterium animalis subsp. lactis BL04) and T4 (ST + La-5 + BL04) and stored at 4±1ºC. Microbiological (population of probiotic bacteria, of the starter and of contaminants), instrumental texture, physical-chemical and sensory (acceptability test) parameters were evaluated. The sensory evaluation was carried out after 7 and 14 days of storage of the products at 4±1ºC, and further analyses were carried out after 1, 7, 14, 21 and 28 days of storage. The populations of the probiotics always remained above the minimum level recommended for a probiotic food, having varied between 7.22 and 7.60 log CFU/g, and between 8.72 and 8.56 log CFU/g during storage, respectively, for L. acidophilus and B. animalis subsp. lactis. The starter culture presented populations between 9.20 and 9.61 log CFU/g in the same period. Coliforms, yeasts and molds presented maximum populations of, respectively, 2.79 and 3.60 log CFU/g. The pH values decreased throughout storage, due to the activity of the acidifying microorganisms present. The moisture ranged from 67.16% up to 70.26% in the same period. All the cheeses studied presented a decrease in hardness and adhesiveness throughout storage. Both after 7 and 14 days, cheese T4 presented the highest sensory acceptance for all attributes evaluated and differed significantly from the other cheeses (p<0.05). After 14 days of storage, cheese T3 presented the lowest acceptance and differed significantly from the other cheeses (p<0.05). The supplementation of petit-suisse cheese T4 with both L. acidophilus and B. animalis subsp. lactis in coculture with a starter culture resulted in a product with excellent market potential, as it presented favorable sensory, microbiological and texture features.

A possibilidade de se obter um produto com grande aceitabilidade nacional, principalmente voltado ao público infantil, com propriedades microbiológicas, nutricionais, físico-químicas e organolépticas ótimas associadas aos benefícios creditados aos probióticos é bastante promissora. Assim, o presente trabalho teve como objetivo verificar a influência do emprego de culturas probióticas compostas pelos microrganismos Lactobacillus acidophilus La-5 e Bifidobacterium animalis subsp. lactis BL04 isolados e em co-cultura, em queijo tipo petit-suisse processado com a adição de Streptococcus thermophilus como cultura starter, sobre as características do produto, ao longo do seu armazenamento refrigerado. Quatro tratamentos de queijo petit-suisse foram estudados (em triplicata): T1 (controle - Streptococcus thermophilus ST), T2 (ST + Lactobacillus acidophilus La-5), T3 (ST + B. animalis subsp. lactis BL04) e T4 (ST + La-5 + BL04) e armazenados a 4±1ºC. Foram avaliados parâmetros microbiológicos (população de probióticos, starter e contaminantes), de textura, físico-químicos e sensoriais (aceitabilidade). As análises sensoriais foram realizadas após 7 e 14 dias e as demais análises, após 1, 7, 14, 21 e 28 dias de armazenamento dos produtos a 4±1oC. As populações dos probióticos estiveram sempre superiores ao mínimo recomendado para um alimento probiótico, tendo variado de 7,22 a 7,60 log UFC/g e de 8,56 a 8,72 log UFC/g durante o armazenamento, para L. acidophilus e B. animalis subsp. lactis, respectivamente. A cultura starter apresentou populações entre 9,20 e 9,61 log UFC/g no mesmo período. As populações máximas para coliformes totais, bolores e leveduras foram de 2,79 e 3,60 log UFC/g, respectivamente. Os valores de pH diminuíram ao longo do armazenamento, devido à atividade dos microrganismos acidificantes presentes. A umidade variou entre 67,16% e 70,26% no mesmo período. Todos os queijos apresentaram queda na dureza e adesividade ao longo do armazenamento. Na análise sensorial, o queijo T4, com a co-cultura, revelou uma aceitabilidade significativamente maior em todos os atributos avaliados (p<0,05). Após 14 dias de armazenamento, o queijo T3 apresentou uma aceitabilidade significativamente inferior aos demais (p<0,05). A suplementação de queijo petit-suisse T4 com Lactobacillus acidophilus La-5 e B. animalis subsp. lactis BL04 em co-cultura com a cultura starter resultou em um produto com excelente potencial comercial, tendo apresentado características sensoriais, microbiológicas e de textura bastante vantajosas. / The possibility of obtaining a product with wide acceptability in Brazil, and which is mainly targeted at children and has excellent microbiological, nutritional, physical-chemical and organoleptic properties associated with the benefits ascribed to probiotic microorganisms is rather promising. Thus, the aim of the present work was to examine the influence of the probiotic cultures of Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BL04, isolated and in co-culture, on the characteristics of petit-suisse cheese produced with the addition of Streptococcus thermophilus as a starter culture, throughout the period of refrigerated storage. Four petit-suisse cheese trials were studied (in triplicates): T1 (control - Streptococcus thermophilus ST), T2 (ST + Lactobacillus acidophilus La-5), T3 (ST + Bifidobacterium animalis subsp. lactis BL04) and T4 (ST + La-5 + BL04) and stored at 4±1ºC. Microbiological (population of probiotic bacteria, of the starter and of contaminants), instrumental texture, physical-chemical and sensory (acceptability test) parameters were evaluated. The sensory evaluation was carried out after 7 and 14 days of storage of the products at 4±1ºC, and further analyses were carried out after 1, 7, 14, 21 and 28 days of storage. The populations of the probiotics always remained above the minimum level recommended for a probiotic food, having varied between 7.22 and 7.60 log CFU/g, and between 8.72 and 8.56 log CFU/g during storage, respectively, for L. acidophilus and B. animalis subsp. lactis. The starter culture presented populations between 9.20 and 9.61 log CFU/g in the same period. Coliforms, yeasts and molds presented maximum populations of, respectively, 2.79 and 3.60 log CFU/g. The pH values decreased throughout storage, due to the activity of the acidifying microorganisms present. The moisture ranged from 67.16% up to 70.26% in the same period. All the cheeses studied presented a decrease in hardness and adhesiveness throughout storage. Both after 7 and 14 days, cheese T4 presented the highest sensory acceptance for all attributes evaluated and differed significantly from the other cheeses (p<0.05). After 14 days of storage, cheese T3 presented the lowest acceptance and differed significantly from the other cheeses (p<0.05). The supplementation of petit-suisse cheese T4 with both L. acidophilus and B. animalis subsp. lactis in coculture with a starter culture resulted in a product with excellent market potential, as it presented favorable sensory, microbiological and texture features.

O objetivo do presente estudo clínico randomizado foi avaliar in vivo, por meio da técnica de biologia molecular Checkerboard DNA-DNA Hybridization: 1) A contaminação de bráquetes metálicos por 4 espécies bacterianas de microrganismos cariogênicos (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei e Lactobacillus acidophilus); e 2) A eficácia da utilização do gluconato de clorexidina a 0,12% (Periogard&reg;) sob a forma de bochechos, sobre esses microrganismos. Participaram do estudo 39 pacientes de 11 a 33 anos de idade, em tratamento com aparelho ortodôntico fixo, nos quais foram colados 2 bráquetes metálicos novos, nos pré-molares. Os pacientes do Grupo Controle (n=20) foram orientados a fazer 2 bochechos semanais com solução placebo, durante 30 dias. Os pacientes do Grupo Experimental (n=19) foram orientados a fazer bochechos com solução à base de gluconato de clorexidina a 0,12% (Periogard&reg;), da mesma forma que o grupo Controle. Decorridos 30 dias, os 2 bráquetes foram removidos de cada paciente e processados para detecção das 4 cepas bacterianas, pela técnica Checkerboard DNADNA Hybridization. Os resultados obtidos foram analisados por meio do teste nãoparamétrico de Kruskal-Wallis, utilizando o software SAS (Statistical Analysis System). O nível de significância adotado foi de 5%. De acordo com os resultados obtidos, observou-se que S. mutans, S. sobrinus, L. casei e L. acidophilus foram detectados em 100% das amostras dos bráquetes de ambos os grupos. No entanto, os bráquetes do grupo Controle encontravam-se mais densamente contaminados por S. mutans e S. sobrinus (p<0,01). No grupo Experimental, embora as quantidades de S. mutans, S. sobrinus, L. casei e L. acidophilus tenham sofrido redução numérica após o uso dos bochechos com solução de gluconato de clorexidina a 0,12%, a comparação com os valores observados no grupo Controle evidenciou diferença estatisticamente significante apenas para S. mutans (p=0,03). Concluiu-se que os bochechos com solução de gluconato de clorexidina a 0,12% podem ser úteis, na prática clínica, com a finalidade de reduzir os níveis de microrganismos cariogênicos, em pacientes portadores de aparelhos ortodônticos fixos. / The purpose of this randomized clinical study was to evaluate in vivo, by the checkerboard DNA-DNA hybridization biomolecular technique: 1) The contamination of metallic brackets by four cariogenic bacterial strains (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei and Lactobacillus acidophilus); and 2) The efficacy of 0.12% chlorhexidine gluconate mouthrinses (Periogard&reg;) against these microorganisms. Thirty-nine 11-33-year-old patients under treatment with fixed orthodontic appliances were enrolled in the study and had 2 new metallic brackets bonded to the premolars. For the patients of the control group (n=20), mouthrinses with a placebo solution were prescribed twice a week during 30 days. The patients randomized to the experimental group (n=19) were instructed to use a 0.12% chlorhexidine gluconate solution (Periogard&reg;) as an oral rinse twice a week during 30 days, in the same way as in control group. After this period, the 2 brackets were removed from each patient and processed for detection of the 4 bacterial strains by checkerboard DNA-DNA hybridization. The obtained data were analyzed statistically by the non-parametric Kruskal-Wallis test using the SAS (Statistical Analysis System) software. A level of significance of 5% was set for all analysis. S. mutans, S. sobrinus, L. casei and L. acidophilus were detected in 100% of the samples from brackets of both groups. However, the brackets of the control group were more heavily contaminated by S. mutans and S. sobrinus (p<0.01). In the experimental group, although S. mutans, S. sobrinus, L. casei and L. acidophilus counts decreased after rinsing with the 0.12% chlorhexidine gluconate solution, the comparison with the values obtained in the control group showed statistically significant difference only for S. mutans (p=0.03). In conclusion, the use of 0.12% chlorhexidine gluconate mouthrinses can be useful in clinical practice to reduce the levels of cariogenic microorganisms in patients under treatment with fixed orthodontic appliances.