Effects of Nutrient Density and exogenous enzymesin Starter Diet on Performance, Intestinal Microflora, Gut Morphology and Immune Response of Broiler Chickens

ABSTRACT A total of 810 one-day-old, straight-run broilers were used to evaluate the effects of dietary nutrient density and feed additives included in the starter diet on their performance, intestinal microbiota, gut morphology, and immune response. A 3×3 factorial arrangement with three nutrient densities (100, 103.75 and 107.5%, as recommended) and three feed additives (no additives, 0.5 g/kg diet Maxi-Gen, and Maxi-Gen + Superzyme + Bio-Phytase at the rate of 0.5, 0.25 and 0.1 g/kg diet, respectively), fed from 1 to 10 d of age. Similar commercial corn-soy grower and finisher diets fed to all birds from 10-24 and 24-42 d of age, respectively. There was higher (p 0.05) body weight gain and lower (p 0.05) feed conversion ratio in chicks fed starter diet with 107.5% nutrient density and Maxi-Gen with or without exogenous enzymes compared with those fed control diet at 10 and 42 d of age. Lactobacilli and Bifidobacteria counts in the cecal content were increased linearly as dietary nutrient density increased in 10-d-old birds (p 0.05). Higher duodenal and jejunal villus height and villus height to crypt depth ratio (p 0.05) were measured in the birds fed the starter diets with 103.75% and 107.5% nutrient density at 5 and 10 d of age. Total anti-SRBC and IgM titers were significantly higher in the broilers fed the 107.5% nutrient density diet containing feed additives at 35 day of age. It is concluded that higher nutrient density and the inclusion of feed additives in the starter diet may improve the growth performance, gut morphology, and immune response of broiler chickens.(AU)

Effects of Nutrient Density and exogenous enzymesin Starter Diet on Performance, Intestinal Microflora, Gut Morphology and Immune Response of Broiler Chickens

ABSTRACT A total of 810 one-day-old, straight-run broilers were used to evaluate the effects of dietary nutrient density and feed additives included in the starter diet on their performance, intestinal microbiota, gut morphology, and immune response. A 3×3 factorial arrangement with three nutrient densities (100, 103.75 and 107.5%, as recommended) and three feed additives (no additives, 0.5 g/kg diet Maxi-Gen, and Maxi-Gen + Superzyme + Bio-Phytase at the rate of 0.5, 0.25 and 0.1 g/kg diet, respectively), fed from 1 to 10 d of age. Similar commercial corn-soy grower and finisher diets fed to all birds from 10-24 and 24-42 d of age, respectively. There was higher (p 0.05) body weight gain and lower (p 0.05) feed conversion ratio in chicks fed starter diet with 107.5% nutrient density and Maxi-Gen with or without exogenous enzymes compared with those fed control diet at 10 and 42 d of age. Lactobacilli and Bifidobacteria counts in the cecal content were increased linearly as dietary nutrient density increased in 10-d-old birds (p 0.05). Higher duodenal and jejunal villus height and villus height to crypt depth ratio (p 0.05) were measured in the birds fed the starter diets with 103.75% and 107.5% nutrient density at 5 and 10 d of age. Total anti-SRBC and IgM titers were significantly higher in the broilers fed the 107.5% nutrient density diet containing feed additives at 35 day of age. It is concluded that higher nutrient density and the inclusion of feed additives in the starter diet may improve the growth performance, gut morphology, and immune response of broiler chickens.

Interference of different storage temperatures in the dynamics of probiotic Bifidobacterium spp. and Streptococcus thermophilus starter cultures in fermented milk

The variations of temperature during the cold chain can impair the quality of live foods, such as fermented milks. Probiotic bacteria are commonly added to food to provide the consumer with beneficial effects. Nevertheless, the concentration of probiotic in the end products should be elevated to ensure functionality. The aim of this study was to evaluate the viability of probiotic strains of bifidobacteria and starter strain of Streptococcus thermophilus in fermented milks at storage temperatures of 4 and 10ºC, for a period of 28 days. Commercial cultures ofBifidobacterium spp. were added to milk fermented byStreptococcus thermophillusand stored for 28 days at 4 and 10ºC. During this period, bifidobacteria and S. thermophilluscultures were monitored to check their behavior in the evaluated storage conditions. Viable bifidobacteria and S. thermophillus counts showed no significant variation during storage at 4 and 10ºC (p 0.05), indicating that both of these conditions are adequate for maintaining their initial concentrations. The results indicate that the storage conditions usually adopted in sale establishments of dairy products are suitable to maintain bifidobacteria and S. thermophillus cultures in fermented milk.

As variações de temperatura que ocorrem durante a cadeia refrigerada da produção leiteira pode interferir na qualidade de alimentos bioativos, como leites fermentados. Bactérias probióticas são usualmente adicionadas a alimentos, visando a oferecer ao consumidor efeitos benéficos. O objetivo deste trabalho foi avaliar a viabilidade de culturas probióticas de bifidobactérias e de cultura starter de Streptococcus thermophilus em leites fermentados, armazenados nas temperaturas de 4 e 10ºC por um período de 28 dias. Culturas comerciais de Bifidobacterium spp. foram adicionadas ao leite fermentado produzido com Streptococcus thermophillus e estocados por 28 dias a 4 e 10ºC. Nesse período, as culturas deBifidobacterium spp. e Streptococcus thermophillus foram monitoradas, com o objetivo de verificar seus comportamentos nas temperaturas de estocagem testadas. As contagens de ambos os micro-organismos não apresentaram variação significativa ao longo do período de estocagem a 4 e a 10ºC (p 0,05), indicando que as duas temperaturas testadas podem oferecer condições adequadas de conservação desse produto. Os resultados obtidos indicaram que as condições de conservação de alimentos usualmente adotadas para produtos lácteos em estabelecimentos comerciais são adequadas para manter as contagens de bifidobactérias e Streptococcus thermophillus em leites fermentados.

Interference of different storage temperatures in the dynamics of probiotic Bifidobacterium spp. and Streptococcus thermophilus starter cultures in fermented milk

The variations of temperature during the cold chain can impair the quality of live foods, such as fermented milks. Probiotic bacteria are commonly added to food to provide the consumer with beneficial effects. Nevertheless, the concentration of probiotic in the end products should be elevated to ensure functionality. The aim of this study was to evaluate the viability of probiotic strains of bifidobacteria and starter strain of Streptococcus thermophilus in fermented milks at storage temperatures of 4 and 10ºC, for a period of 28 days. Commercial cultures ofBifidobacterium spp. were added to milk fermented byStreptococcus thermophillusand stored for 28 days at 4 and 10ºC. During this period, bifidobacteria and S. thermophilluscultures were monitored to check their behavior in the evaluated storage conditions. Viable bifidobacteria and S. thermophillus counts showed no significant variation during storage at 4 and 10ºC (p 0.05), indicating that both of these conditions are adequate for maintaining their initial concentrations. The results indicate that the storage conditions usually adopted in sale establishments of dairy products are suitable to maintain bifidobacteria and S. thermophillus cultures in fermented milk.

As variações de temperatura que ocorrem durante a cadeia refrigerada da produção leiteira pode interferir na qualidade de alimentos bioativos, como leites fermentados. Bactérias probióticas são usualmente adicionadas a alimentos, visando a oferecer ao consumidor efeitos benéficos. O objetivo deste trabalho foi avaliar a viabilidade de culturas probióticas de bifidobactérias e de cultura starter de Streptococcus thermophilus em leites fermentados, armazenados nas temperaturas de 4 e 10ºC por um período de 28 dias. Culturas comerciais de Bifidobacterium spp. foram adicionadas ao leite fermentado produzido com Streptococcus thermophillus e estocados por 28 dias a 4 e 10ºC. Nesse período, as culturas deBifidobacterium spp. e Streptococcus thermophillus foram monitoradas, com o objetivo de verificar seus comportamentos nas temperaturas de estocagem testadas. As contagens de ambos os micro-organismos não apresentaram variação significativa ao longo do período de estocagem a 4 e a 10ºC (p 0,05), indicando que as duas temperaturas testadas podem oferecer condições adequadas de conservação desse produto. Os resultados obtidos indicaram que as condições de conservação de alimentos usualmente adotadas para produtos lácteos em estabelecimentos comerciais são adequadas para manter as contagens de bifidobactérias e Streptococcus thermophillus em leites fermentados.

Interference of different storage temperatures in the dynamics of probiotic Bifidobacterium spp. and Streptococcus thermophilus starter cultures in fermented milk / Interferência de diferentes temperaturas de armazenamento na dinâmica de Bifidobacterium spp. probióticas e cultura starterStreptococcus thermophilus em leite fermentado

The variations of temperature during the cold chain can impair the quality of live foods, such as fermented milks. Probiotic bacteria are commonly added to food to provide the consumer with beneficial effects. Nevertheless, the concentration of probiotic in the end products should be elevated to ensure functionality. The aim of this study was to evaluate the viability of probiotic strains of bifidobacteria and starter strain of Streptococcus thermophilus in fermented milks at storage temperatures of 4 and 10ºC, for a period of 28 days. Commercial cultures of Bifidobacterium spp. were added to milk fermented by Streptococcus thermophillus and stored for 28 days at 4 and 10ºC. During this period, bifidobacteria and S. thermophilluscultures were monitored to check their behavior in the evaluated storage conditions. Viable bifidobacteria and S. thermophillus counts showed no significant variation during storage at 4 and 10ºC (p < 0.05), indicating that both of these conditions are adequate for maintaining their initial concentrations. The results indicate that the storage conditions usually adopted in sale establishments of dairy products are suitable to maintain bifidobacteria and S. thermophillus cultures in fermented milk.(AU)

As variações de temperatura que ocorrem durante a cadeia refrigerada da produção leiteira pode interferir na qualidade de alimentos bioativos, como leites fermentados. Bactérias probióticas são usualmente adicionadas a alimentos, visando a oferecer ao consumidor efeitos benéficos. O objetivo deste trabalho foi avaliar a viabilidade de culturas probióticas de bifidobactérias e de cultura starter de Streptococcus thermophilus em leites fermentados, armazenados nas temperaturas de 4 e 10ºC por um período de 28 dias. Culturas comerciais de Bifidobacterium spp. foram adicionadas ao leite fermentado produzido com Streptococcus thermophillus e estocados por 28 dias a 4 e 10ºC. Nesse período, as culturas de Bifidobacterium spp. e Streptococcus thermophillus foram monitoradas, com o objetivo de verificar seus comportamentos nas temperaturas de estocagem testadas. As contagens de ambos os micro-organismos não apresentaram variação significativa ao longo do período de estocagem a 4 e a 10ºC (p < 0,05), indicando que as duas temperaturas testadas podem oferecer condições adequadas de conservação desse produto. Os resultados obtidos indicaram que as condições de conservação de alimentos usualmente adotadas para produtos lácteos em estabelecimentos comerciais são adequadas para manter as contagens de bifidobactérias e Streptococcus thermophillus em leites fermentados.(AU)

Interference of different storage temperatures in the dynamics of probiotic Bifidobacterium spp. and Streptococcus thermophilus starter cultures in fermented milk / Interferência de diferentes temperaturas de armazenamento na dinâmica de Bifidobacterium spp. probióticas e cultura starter Streptococcus thermophilus em leite fermentado

The variations of temperature during the cold chain can impair the quality of live foods, such as fermented milks. Probiotic bacteria are commonly added to food to provide the consumer with beneficial effects. Nevertheless, the concentration of probiotic in the end products should be elevated to ensure functionality. The aim of this study was to evaluate the viability of probiotic strains of bifidobacteria and starter strain of Streptococcus thermophilus in fermented milks at storage temperatures of 4 and 10C, for a period of 28 days. Commercial cultures of Bifidobacterium spp. were added to milk fermented by Streptococcus thermophillus and stored for 28 days at 4 and 10ºC. During this period, bifidobacteria and S. thermophillus cultures were monitored to check their behavior in the evaluated storage conditions. Viable bifidobacteria and S. thermophillus counts showed no significant variation during storage at 4 and 10ºC (p 0.05), indicating that both of these conditions are adequate for maintaining their initial concentrations. The results indicate that the storage conditions usually adopted in sale establishments of dairy products are suitable to maintain bifidobacteria and S. thermophillus cultures in fermented milk.

As variações de temperatura que ocorrem durante a cadeia refrigerada da produção leiteira pode interferir na qualidade de alimentos bioativos, como leites fermentados. Bactérias probióticas são usualmente adicionadas a alimentos, visando a oferecer ao consumidor efeitos benéficos. O objetivo deste trabalho foi avaliar a viabilidade de culturas probióticas de bifidobactérias e de cultura starter de Streptococcus thermophilus em leites fermentados, armazenados nas temperaturas de 4 e 10C por um período de 28 dias. Culturas comerciais de Bifidobacterium spp. foram adicionadas ao leite fermentado produzido com Streptococcus thermophillus e estocados por 28 dias a 4 e 10C. Nesse período, as culturas de Bifidobacterium spp. e Streptococcus thermophillus foram monitoradas, com o objetivo de verificar seus comportamentos nas temperaturas de estocagem testadas. As contagens de ambos os micro-organismos não apresentaram variação significativa ao longo do período de estocagem a 4 e a 10C (p 0,05), indicando que as duas temperaturas testadas podem oferecer condições adequadas de conservação desse produto. Os resultados obtidos indicaram que as condições de conservação de alimentos usualmente adotadas para produtos lácteos em estabelecimentos comerciais são adequadas para manter as contagens de bifidobactérias e Streptococcus thermophillus em leites fermentados.

Interference of different storage temperatures in the dynamics of probiotic Bifidobacterium spp. and Streptococcus thermophilus starter cultures in fermented milk / Interferência de diferentes temperaturas de armazenamento na dinâmica de Bifidobacterium spp. probióticas e cultura starter Streptococcus thermophilus em leite fermentado

The variations of temperature during the cold chain can impair the quality of live foods, such as fermented milks. Probiotic bacteria are commonly added to food to provide the consumer with beneficial effects. Nevertheless, the concentration of probiotic in the end products should be elevated to ensure functionality. The aim of this study was to evaluate the viability of probiotic strains of bifidobacteria and starter strain of Streptococcus thermophilus in fermented milks at storage temperatures of 4 and 10C, for a period of 28 days. Commercial cultures of Bifidobacterium spp. were added to milk fermented by Streptococcus thermophillus and stored for 28 days at 4 and 10ºC. During this period, bifidobacteria and S. thermophillus cultures were monitored to check their behavior in the evaluated storage conditions. Viable bifidobacteria and S. thermophillus counts showed no significant variation during storage at 4 and 10ºC (p 0.05), indicating that both of these conditions are adequate for maintaining their initial concentrations. The results indicate that the storage conditions usually adopted in sale establishments of dairy products are suitable to maintain bifidobacteria and S. thermophillus cultures in fermented milk.(AU)

As variações de temperatura que ocorrem durante a cadeia refrigerada da produção leiteira pode interferir na qualidade de alimentos bioativos, como leites fermentados. Bactérias probióticas são usualmente adicionadas a alimentos, visando a oferecer ao consumidor efeitos benéficos. O objetivo deste trabalho foi avaliar a viabilidade de culturas probióticas de bifidobactérias e de cultura starter de Streptococcus thermophilus em leites fermentados, armazenados nas temperaturas de 4 e 10C por um período de 28 dias. Culturas comerciais de Bifidobacterium spp. foram adicionadas ao leite fermentado produzido com Streptococcus thermophillus e estocados por 28 dias a 4 e 10C. Nesse período, as culturas de Bifidobacterium spp. e Streptococcus thermophillus foram monitoradas, com o objetivo de verificar seus comportamentos nas temperaturas de estocagem testadas. As contagens de ambos os micro-organismos não apresentaram variação significativa ao longo do período de estocagem a 4 e a 10C (p 0,05), indicando que as duas temperaturas testadas podem oferecer condições adequadas de conservação desse produto. Os resultados obtidos indicaram que as condições de conservação de alimentos usualmente adotadas para produtos lácteos em estabelecimentos comerciais são adequadas para manter as contagens de bifidobactérias e Streptococcus thermophillus em leites fermentados.(AU)

Lactobacillus Pentosus Ita23 and L. Acidipiscis Ita44 Enhance Feed Conversion Efficiency and Beneficial Gut Microbiota in Broiler Chickens

Although the use of probiotics especially Lactobacillus and Bifidobacteria species on growth and feed utilization in poultry production has been extensively studied, the results were inconsistent presumably because the mode of action of probiotic is multi-factorial and each probiotic strain may affect the host in a specific manner. This study investigated the probiotic effect of two strains of Lactobacillus (Lactobacillus pentosus ITA23 and Lactobacillus acidophilus ITA44; 109 cells/kg feed) isolated from mulberry (Morus Alba) silage on the growth performance, cecal microbial population, and blood cholesterol of broiler chickens. One hundred twenty male broiler chicks (Cobb500) were randomLy allocated to two groups (control and treatment) of six replicates (cages) with 10 chicks per cage. Chicks in the control group received a standard diet and those in the treatment group received the same diet supplemented with 109 cells of the above Lactobacillus per kg feed. Supplementation of Lactobacillus did not affect body weight gain (averaged 1604 g at 35 days old) but feed conversion ratio improved (p 0.01) by 6.4% due to reduction in feed intake (p 0.01) by birds in the treatment group. Supplementation also increased the population of Lactobacillus spp. and reduced pathogens E. coli in the cecal samples. Although Lactobacillus supplementation tends to reduce serum total cholesterol, low-density lipoprotein (LDL) and triglyceride concentrations, these values were not significantly different from those of the control group. Results of this study showed that L. pentosus ITA23 and L. acidophilus ITA44 are potential probiotics to be used in poultry diets.(AU)

Lactobacillus Pentosus Ita23 and L. Acidipiscis Ita44 Enhance Feed Conversion Efficiency and Beneficial Gut Microbiota in Broiler Chickens

Although the use of probiotics especially Lactobacillus and Bifidobacteria species on growth and feed utilization in poultry production has been extensively studied, the results were inconsistent presumably because the mode of action of probiotic is multi-factorial and each probiotic strain may affect the host in a specific manner. This study investigated the probiotic effect of two strains of Lactobacillus (Lactobacillus pentosus ITA23 and Lactobacillus acidophilus ITA44; 109 cells/kg feed) isolated from mulberry (Morus Alba) silage on the growth performance, cecal microbial population, and blood cholesterol of broiler chickens. One hundred twenty male broiler chicks (Cobb500) were randomLy allocated to two groups (control and treatment) of six replicates (cages) with 10 chicks per cage. Chicks in the control group received a standard diet and those in the treatment group received the same diet supplemented with 109 cells of the above Lactobacillus per kg feed. Supplementation of Lactobacillus did not affect body weight gain (averaged 1604 g at 35 days old) but feed conversion ratio improved (p 0.01) by 6.4% due to reduction in feed intake (p 0.01) by birds in the treatment group. Supplementation also increased the population of Lactobacillus spp. and reduced pathogens E. coli in the cecal samples. Although Lactobacillus supplementation tends to reduce serum total cholesterol, low-density lipoprotein (LDL) and triglyceride concentrations, these values were not significantly different from those of the control group. Results of this study showed that L. pentosus ITA23 and L. acidophilus ITA44 are potential probiotics to be used in poultry diets.

Evaluation of culture media for selective enumeration of bifidobacteria and lactic acid bacteria

The purpose of this study was to test the suitability of Transgalactosylated oligosaccharides-mupirocin lithium salt (TOS-MUP) and MRS-clindamycin-ciprofloxacin (MRS-CC) agars, along with several other culture media, for selectively enumerating bifidobacteria and lactic acid bacteria (LAB) species commonly used to make fermented milks. Pure culture suspensions of a total of 13 dairy bacteria strains, belonging to eight species and five genera, were tested for growth capability under various incubation conditions. TOS-MUP agar was successfully used for the selective enumeration of both Bifidobacterium animalis subsp. lactis BB-12 and B. breve M-16 V. MRS-CC agar showed relatively good selectivity for Lactobacillus acidophilus, however, it also promoted the growth of Lb. casei strains. For this reason, MRS-CC agar can only be used as a selective medium for the enumeration of Lb. acidophilus if Lb. casei is not present in a product at levels similar to or exceeding those of Lb. acidophilus. Unlike bifidobacteria and coccus-shaped LAB, all the lactobacilli strains involved in this work were found to grow well in MRS pH 5.4 agar incubated under anaerobiosis at 37 °C for 72 h. Therefore, this method proved to be particularly suitable for the selective enumeration of Lactobacillus spp.

PRODUÇÃO E PURIFICAÇÃO DE FRUTOSILTRANSFERASE POR Aspergillus Flavus UTILIZANDO BORRA DE CAFÉ (Coffea sp.) ATRAVÉS DA FERMENTAÇÃO EM ESTADO SÓLIDO

Frutosiltransferase (FTase) é uma enzima que catalisa a quebra de ligações glicosídicas da molécula de sacarose transferência do grupo frutosil para uma outra molécula, podendo ser sacarose ou frutooligossacarídeos (FOS), ocorrendo a liberação de 6 glicose. O FOS é um prebiótico convencional que apresenta um baixo valor calorimétrico e promove a estimulação seletiva do crescimento microbiano intestinal, especialmente de bifidobactérias e lactobacilos, reduzindo os riscos de doenças cardiovasculares, câncer de cólon e obesidade. No entanto para obtenção a utilização de processos enzimáticos é mais caro devido ao alto custo de produção e baixa estabilidade das enzimas. Contudo esses fatores podem ser contornados com a utilização de microrganismos e resíduos agroindustriais como um meio de cultivo para uma fermentação solida, tornando o procedimento mais econômico e viável. O objetivo desse trabalho foi à produção da enzima frutosiltransferase em fermentação solida utilizando borra de café a partir de Aspergillus flavus, onde foi possível ainda verificar que 72h de crescimento apresentou maior produção da enzima com atividade especifica de 31,5 U/mg nas seguintes condições, 60% de umidade, concentração de esporos 106 esporos/mL a 30°C durante 120 horas. O extrato bruto contendo a enzima foi submetido a uma seleção de precipitação entre a cetona, sulfato de amônia e etanol, onde a maior atividade enzimática se deu utilizando a precipitação com acetona 161,85 U/mL e fator de purificação relativamente alto de 10,70. Posteriormente a amostra foi purificada através de sistema de cromatografia de troca iônica DEAE SEPHADEX A50 e SUPERDEX AKTA nas colunas DEAE-sephadex sistema haithap e superdex 75 sendo realizadas em sequência. A enzima purificada apresentou peso molecular de 57 kDa em Superdex G-75. Quanto a caracterização, apresentou temperatura ótima de 60ºC respectivamente, obtendo termo estabilidade a 50ºC. Os resultados da purificação mostraram uma fração com atividade enzimática para frutosiltransferase pura na qual obtive um fator de purificação de 86,16. Quando observado sua síntese enzimática foi constatado que a enzima consegue produzir frutooligossacarideos, sendo eles kestose e nistose. Os resultados obtidos no presente estudo mostram o potencial promissor da frutosiltransferase produzida por Aspergillus flavus e na sua utilização para produção de frutooligossacarideos.

Frutosiltransferase (FTase) is an enzyme that catalyzes the breakdown of the glycosidic bonds of the sucrose molecule by transferring the fructosyl group to another molecule, which can be sucrose or fructooligosaccharides (FOS), resulting in the release of glucose. FOS is a conventional prebiotic that has a low calorimetric value and promotes the selective stimulation of the intestinal microbial growth, especially of bifidobacteria and lactobacilli, reducing the risks of cardiovascular disease, colon cancer and obesity. However to obtain the use of enzymatic processes is more expensive due to the high cost of production. However, these factors can be overcome with the use of microorganisms and agroindustrial residues as a culture medium for a solid fermentation, making the procedure more economical and feasible. The objective of this work was the production of the enzyme fructosyltransferase in solid fermentation using coffee grounds from Aspergillus flavus, where it was possible to verify that 72h of growth presented higher production of the enzyme with specific activity of 31.5 U / mg in the following conditions, 60% moisture, spore concentration 106 spores / mL at 30 ° C for 120 hours. The crude extract containing the enzyme was subjected to a precipitation selection between ketone, ammonium sulfate and ethanol, where the highest enzymatic activity was given using the precipitation with acetone 161.85 U / mL and relatively high purification factor of 10.70. Subsequently the sample was purified by DEAE SEPHADEX A50 and SUPERDEX AKTA ion exchange chromatography systems on the DEAE-sephadex system haithap and superdex 75 columns being carried out in sequence. The purified enzyme had 57 kDa molecular weight in Superdex G-75. Regarding the characterization, it presented optimum temperature of 60ºC respectively, obtaining thermo stability at 50ºC. The purification results showed a fraction with enzymatic activity for pure fructosyltransferase in which it obtained a purification factor of 86.16. When observed its enzymatic synthesis it was verified that the enzyme can produce fructooligosaccharides, being kestose and nystose. The results obtained in the present study show the promising potential of the fructosyltransferase produced by Aspergillus flavus and its use for the production of fructooligosaccharides.

Effect of probiotics on the development of dimethylhydrazine-induced preneoplastic lesions in the mice colon

PURPOSE: To determine the effect of probiotics on the development of chemically induced (1, 2-dimethylhydrazine) colonic preneoplastic lesions, in mice. METHODS: The animals were divided into five groups. The control group was injected with carcinogen alone and the other groups also received probiotics (1- Lactobacillus delbrueckii UFV-H2b20; 2- Bifidobacterium animalis var. lactis Bb12; 3- L. delbrueckii UFV-H2b20 plus B. animalis var. lactis Bb12; and 4- Saccharomyces boulardii) administered orally in drinking water throughout fourteen weeks. RESULTS: Consumption of lactobacilli and bifidobacteria alone resulted in a significant reduction of the total number of aberrant crypt foci (55.7% and 45.1%, respectively). Significant reduction in the number of these small foci (<3 aberrant crypts) was only observed in the group treated with lactobacilli (52.2%) in comparison to control group. The number of larger foci (>3 aberrant crypts) crypts had no significant reduction. CONCLUSION: L. delbrueckii UFV-H2b20 and B. animalis var. lactis Bb12 administered alone protect colonic preneoplastic lesions in mice, while the combined treatment of these bacteria and the administration of S.boulardii were not effective in reducing such colonic lesions.(AU)

Suplemento proteico fermentado para abelhas Apis mellifera africanizadas

O objetivo desta pesquisa foi desenvolver um método com condições ideais para a fermentação de um suplemento proteico que exerça efeito positivo sobre a longevidade das abelhas africanizadas, confinadas em gaiola. Os probióticos utilizados para a fermentação do produto foram: Bifidobacterias, Lactobacilos, e Saccharamyces e o prebiótico inulina. Para o desenvolvimento do método, foram realizados o Planejamento Fatorial Fracionado (24-1 + 4pc) e o Delineamento Composto Central Rotacional (22 + 4 axiais + 4 pc), totalizando 24 ensaios. Em um período de fermentação de cinco dias houve a diminuição do pH e aumento de ácido lático. A maior quantidade de bactérias ácidoláticas presentes foi de 109 UFC/g de suplemento. Temperaturas baixas e umidades altas produziram os melhores efeitos sobre as respostas fermentativas. A condição ideal que possibilitou eficiente fermentação foi: 38,96ºC de temperatura, 66,00% de umidade, 2,00% de inulina e 10,00% de probióticos. Para testar o efeito do suplemento fermentado por probióticos em abelhas africanizadas, foram utilizados quatro tratamentos com cinco repetições. Os tratamentos foram: xarope, pólen, suplemento proteico e suplemento proteico fermentado. As respostas avaliadas foram: consumo do suplemento, taxa de mortalidade e aumento da longevidade. Observou-se em abelhas alimentadas com suplemento proteico fermentado por probióticos: menor consumo médio, menor taxa de mortalidade e a maior longevidade. Foi possível criar um ambiente ideal que permitiu a fermentação do suplemento proteico em apenas cinco dias. A fermentação e a presença de probióticos promoveram nas abelhas aumento da longevidade em sistema fechado.

This research was carried out to develop a method with ideal conditions for fermentation of a protein supplement that promotes a positive effect on longevity of Africanized honeybees confined in cages. The probiotics used for fermentation were: Bifidobacteria, Lactobacilli, and Saccharomyces and the inulin prebiotic. For the method development, Fractional Factorial Planning (24-1 + 4pc) and Rotational Central Compound Design (22 + 4 axial + 4 ch) were performed, totaling 24 trials. In a fermentation period of five days there was a decrease in the pH and an increase in the lactic acid. The greater amount of lactic acid bacteria present was of 109 CFU/g of supplement. The low temperatures and high humidity produced the best effects on the fermentative responses. The ideal condition that made possible an efficient fermentation was: temperature 38.96°C, humidity of 66.00%, inulin 2.00% and probiotics 10.00%. To try out the effect of the fermented supplement by probiotic on Africanized honeybees, four treatments with five repetitions were used. The treatments were: sugar syrup, pollen, protein supplement and fermented protein supplement. The responses evaluated were: supplement intake, mortality rate and longevity. It was observed in honeybees fed with protein supplement fermented by probiotics the lowest mean intake, lowest mortality rate and greatest increase in longevity. Also, it was possible to create an ideal environment that allowed fermentation of the protein supplement, in just five days. The fermentation and the presence of probiotics promoted in bees the increase of longevity in closed system.

UTILIZAÇÃO DE IMUNOESTIMULANTES EM TILÁPIADO-NILO (Oreochromis niloticus)

O objetivo deste estudo foi avaliar o desempenho, a microbiota intestinal, os parâmetros hematológicos e imunológicos de tilápia-do-nilo alimentados com ração suplementada com probiótico e prebióticos, além da sobrevivência frente a desafio bacteriano com Aeromonas hydrophila. Para o primeiro experimento foram utilizadas 240 tilápias (8,84±1,29g), distribuídas em 24 aquários de 40L, com seis tratamentos, com 4 repetições e alimentadas durante 63 dias: Controle (C), Probiótico DBA, constituído de - 3,5x109UFC g-1 Bifidobacteria, 3,5x109UFC g-1 Lactobacillus acidophilus e 3,5x109UFC g-1 Enterococcus faecium (PRO), os Prébióticos Actigen - MOS (PRE1), Quitosana (PRE2) e os simbióticos DBA e Actigen MOS (S1), DBA e Quitosana (S2). No desafio bacteriano foram utilizadas 180 tilápias-do-nilo, dustribuídas em 18 aquários de 40L, com os mesmos seis tratamentos, com 3 repetições. Os peixes receberam as dietas experimentais durante 21 dias, três vezes ao dia, até a saciedade. Após o período de alimentação os peixes foram infectados via injeção intraperitoneal com 1,26x108 UFC mL-1 de Aeromonas hydrophila e observados durante 15 dias para a detecção da mortalidade. O delineamento dos dois experimentos foi inteiramente casualizado, e em ambos os casos, os dados foram submetidos ao teste de ANOVA e, quando p<0,05, foi realizado o teste de Dunnett. No desempenho zootécnico, houve diferença significativa na biomassa final dos tratamentos S2 e PRE2 quando comparados com S1, e maior ganho de peso individual no grupo S2 em relação ao grupo S1. A avaliação da microbiota intestinal não apresentou diferença estatística (p>0,05) nos índices de bactérias dos gêneros Lactobacillus spp., Bifidobacterium spp. e Enterococcus spp no intestino dos animais que foram suplementados com os prebióticos Actigen e Quitosana e o probiótico DBA, quando utilizados em separado e em conjunto. No desafio bacteriano, o nível de proteção relativa (NPR) foi maior no grupo S1 (40%) em relação a proteção do grupo controle, apontando interação positiva entre o probiótico DBA e o prebiótico mananoligossacarídeo (Actigen). Não foram observadas diferenças significativas nos parâmetros hematológicos e imunológicos.

The aim of this study was to evaluate the growth performance, intestinal microbiota, hematological and immunological parameters, from Nile tilapia fed with supplemented diet with probiotic and prebiotics, and bacterial challenge with Aeromonas hydrophila survival. For the first experiment were used 240 tilapia, distributed in 24 tanks of 40L, with six treatments, 4 replicates and were fed for 63 days: Control (C), Probiotic DBA, consisting of - 3,5x109UFC g-1 Bifidobacteria, 3,5x109UFC g-1 Lactobacillus acidophilus and 3,5x109UFC g-1 Enterococcus faecium (PRO), prebiotics Actigen - MOS (PRE1), chitosan (PRE2), and symbiotic DBA and Actigen MOS (S1), DBA and chitosan (S2). In bacterial challenge were used 180 Nile tilapia, distributed in 18 tanks 40L, with the same six treatments with three repetitions. Fish received the experimental diets for 21 days, three times daily until satiation. After the feeding period the fish were infected by intraperitoneal injection with 1,26x108 UFC mL- 1 from Aeromonas hydrophila and observed for 15 days for the detection of mortality. The design of the two trials was randomized, and in both cases, the data were submitted to ANOVA test and p <0.05, was held the Dunnett test. On the growth performance, there was a significant difference in the final biomass of the S2 and PRE2 treatments compared to S1, and greater individual weight gain in the S2 group compared to the S1 group. The evaluation of the intestinal microbiota showed no statistical difference (p> 0.05) in the levels of bacteria of the genus Lactobacillus spp., Bifidobacterium spp. and Enterococcus spp the intestine of the animals that were supplemented with the prebiotics Actigen and Chitosan and the probiotic DBA, used separately and together.In the bacterial challenge, the level of relative protection (NPR) was higher in the group S1 (40%) compared to the control group protection, pointing to a positive interaction between the DBA probiotic and prebiotic mannan oligosaccharide (Actigen). No significant differences were observed in hematological and immunological parameters.

Selective enumeration and viability of Bifidobacterium animalis subsp. lactis in a new fermented milk product

One of the key focuses of today's dairy industry worldwide is the continued development of new products, especially probiotic-based products. Buttermilk is originally a by-product of butter making fermented by Mesophilic Aromatic Cultures (MAC). It can also be made by fermentation of pasteurized whole milk or skimmed milk. This product is not marketed in Brazil. The objectives of this work were: (1) to develop a selective medium for Bifidobacterium animalis subsp. lactis enumeration and (2) to determine the viability of this microorganism during the shelf life of the buttermilk. Skim milk added with 10% sucrose or 0.03% sucralose was pasteurized and inoculated with a composite starter culture consisting of 1% MAC (containing Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis biovar. diacetylactis and Leuconostoc mesenteroides subsp. cremoris) and 2% Bifidobacterium animalis subsp. lactis. To attain selective counts of Bif. animalis subsp. lactis the MRS agar supplemented with 0.5% L-cysteine hydrochloride at 10%, 1% lithium chloride at 10%, 0.01% aniline blue and 0.5% dicloxacillin at 0.1% was modified by increasing the antibiotic concentration, addition of NaCl, adjusting pH to 4.8 or increasing the incubation temperature (from 37 to 45ºC). Raising the incubation temperature to 45ºC was found to be efficient in inhibiting the MAC cultures, even in media not added with dicloxacillin. Bif. animalis subsp. lactis exhibited high viability in the product. The buttermilk product prepared with sucrose and sweetener contained in excess of 10(8) cfu.ml-1 bifidobacteria throughout the shelf life of the product (28 days).

Atualmente, um dos principais focos da indústria de laticínios em todo o mundo é o desenvolvimento de novos produtos, especialmente probióticos. Buttermilk é originalmente um sub-produto do processamento da manteiga fermentado por Culturas Aromáticas Mesofílicas (MAC). Pode também ser feito pela fermentação de leite integral ou desnatado. Este produto não é comercializado no Brasil. Os objetivos deste trabalho foram o desenvolvimento de meio de cultura seletivo para Bifidobacterium animalis subsp. lactis e a determinação da viabilidade deste microrganismo durante a vida de prateleira do buttermilk produzido. Leite desnatado foi adicionado de 10% da sacarose ou 0,03% de sucralose, pasteurizado e inoculado com 1% de MAC composto por Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis biovar. diacetylactis e Leuconostoc mesenteroides subsp. cremoris e por 2% de Bifidobacterium animalis subsp. lactis. Para obter contagens seletivas de Bif. animalis subsp. lactis, o meio MRS ágar suplementado com 0,5% L-cisteína HCl a 10%, 1% cloreto de lítio a 10%, 0,01% azul de anilina e 0,5% dicloxacilina a 0,1% foi modificado pelo aumento da concentração de antibiótico, adição de NaCl, ajuste de pH para 4,8 ou aumento da temperatura de incubação (de 37 para 45ºC). A temperatura de incubação de 45ºC foi eficiente para inibir as culturas MAC mesmo sem adição de antibiótico ao meio. Bif. animalis subsp. lactis apresentou alta viabilidade no produto. O buttermilk preparado com sacarose e edulcorante, apresentou mais de 10(8) ufc.ml-1 de Bif. animalis subsp. lactis durante a vida-de-prateleira (28 dias).

Emprego de probióticos em infecções gastrointestinais e em terapias com antibióticos / Use of probiotics in gastrointestinal infections and antibiotic therapy

São considerados probióticos alguns micro-organismos, principalmente bactérias, que atuam na regulação da microflora intestinal e conferindo ao hospedeiro alguns benefícios, como a melhoria do sistema imunológico, proteção contra colonização de patógenos e controle da flora intestinal em tratamentos com antibióticos. Esta revisão objetiva discorrer sobre algumas funções destes micro-organismos bem como relatar alguns resultados encontrados em experimentos realizados visando à compreensão a ação dos probióticos em infecções gastrointestinais. Os trabalhos realizados nesta área indicam que os probióticos podem ser considerados como uma possibilidade na prevenção e no tratamento de infecções gastrointestinais, bem como coadjuvante em terapias com antibióticos.(AU)

Probiotics are some microorganisms,mainly bacteria, that act inthe regulation of intestinal microfloraand giving the host some benefitssuch as improved immune system,protection against colonization of pathogensand control of intestinal florain antibiotic treatments. This reviewaims to report some features of thesemicroorganisms as well to repot someresults found in experiments aimed atunderstanding the actin of probioticsin gastrointestinal infections. Theworks done in this area indicate thatprobiotics can be considered as apossibility in preventing and treatinggastrointestinal infections, as well asadjuvant therapies with antibiotics. (AU)

Properties of Streptococcus thermophilus fermented milk containing variable concentrations of Bifidobacterium longum and Lactobacillus acidophilus

Sensory evaluation and analysis of pH, titratable acidity and microbial counts after 1, 7, 14 and 21 days of storage of five combinations of lyophilized Bifidobacterium longum and/or Lactobacillus acidophilus added to milk fermented with Streptococcus thermophilus were studied during storage at 4ºC. The taste and acidity sensory attributes were significantly (P 0.05) perceived as the storage time increased. Favourite combinations contained, initially, Bif. longum and L. acidophilus (10(8) and 10(7) cfu/mL, respectively) and Bif. longum only (10(8) cfu/mL). These combinations did not differ significantly among them and neither from the standard fermented milk. The lowest sensory scores and highest titratable acidity values were obtained for fermented milk added of L. acidophilus (10(8) cfu/mL initial concentration). During the 21 days of storage at 4ºC the viable cell counts of: Str. thermophilus did not change. Bif. longum viable cells counts kept constant or reduced 1 logarithmic cycle and L. acidophilus reduced 1 or 2 logarithmic cycle. We did not observe inhibition of over-acidification caused by the presence of bifidobacteria or L. acidophilus.

Foram estudadas as características sensoriais, de pH e acidez de cinco combinações de leites fermentados por Streptococcus thermophilus, adicionados de Bifidobacterium longum e/ou Lactobacillus acidophilus liofilizados. Os efeitos sobre as características sensoriais tornaram-se significantes (p 0,05) com o aumento do tempo de estocagem. As combinações favoritas continham, inicialmente, Bif. longum e L. acidophilus (10(8) e 10(7)ufc/mL, respectivamente) e Bif. longum apenas (10(8)ufc/mL). Estas combinações não diferiram significativamente do leite fermentado padrão, nem entre si. As notas mais baixas e os maiores valores de acidez titulável foram apresentados pelo leite fermentado por Streptococcus thermophilus contendo apenas L. acidophilus (10(8)ufc/mL, concentração inicial). Durante 21 dias de estocagem a 4ºC, as contagens de células viáveis de Str. thermophilus não mudaram, as de Bif. longum mantiveram-se constantes ou reduziram em um ciclo logarítmico e, as de L. acidophilus reduziram de 1 a 2 ciclos logarítmicos. Não foi observada inibição da pós-acidificação resultante da presença de bifidobactéria ou L. acidophilus.

Efeito da adição de parede celular de levedura sobre a digestibilidade, microbiota, ácidos graxos de cadeia curta e aminas fecais e parâmetros hematológicos e imunológicos de cães

Considerando a importância da eubiota intestinal na saúde dos animais, o papel da dieta em sua composição e metabolismo tem sido estudado. Neste aspecto os prebióticos têm merecido destaque, incluindo-se a parede celular de leveduras (PCL), uma fonte natural de mananoligossacarídeos. Para o estudo de seu potencial prebiótico para cães, foram empregadas quatro dietas isonutrientes com inclusão de 0% (T0), 0,15% (T15), 0,30% (T30) e 0,45% (T45) de PCL. Foram utilizados oito Beagles adultos, divididos em dois quadrados latinos 4 X 4. Em cada período uma fase de adaptação de 15 dias precedeu 5 dias de coleta total de fezes para o ensaio de digestibilidade e 1 dia de coleta de fezes frescas para enumeração bacteriana, mensuração de pH e determinação das concentrações de ácidos graxos de cadeia curta e aminas bioativas. No dia 21 de cada período, amostra de sangue foi colhida para quantificação imunofenotípica por citometria de fluxo das subpopulações linfocitárias CD5+CD4+, CD5+, CD5+CD8+, CD45+ e CD45+CD21+. Os dados foram avaliados pelo Proc GLM do SAS, sendo as médias comparadas pelo teste de Tukey e por contraste polinomial e ortogonal (p<0,1). Os coeficientes de digestibilidade dos nutrientes não variaram com a inclusão de PCL (p>0,1). A inclusão de PCL não resultou em diferenças quanto às contagens de anaeróbios totais, aeróbios totais, bifidobactéria, lactobacilos, clostridios nas fezes dos cães (p>0.1). Verificou-se aumento linear na concentração fecal de butirato (p=0,055), redução linear das concentrações fecais de tiramina (P=0,1) e histamina (P=0,07) e redução quadrática de feniletilamina e triptamina (P=0,07). Os cães apresentaram, também, aumento linear na concentração de linfócitos CD5+ (P=0,10) e maior número de linfócitos CD45+CD21+ (P=0,053) mediante ingestão de PCL. Concluiu-se que a PCL apresenta efeito prebiótico...

Considering the importance of the intestinal microbiota to the animal health, the role of diet upon microbiota composition and metabolism have been studied. Prebiotics have been emphasized on this regard, including the yeasts cell walls (YCW), a natural source of mannanoligosaccharides. To study the prebiotic potential of this compound for dogs, four isonutrient diets were used with inclusion of 0% (T0), 0,15% (T15), 0,30% (T30) and 0,45% (T45) of YCW. Eight Beagles adults were used, arranged in two 4 X 4 Latin squares. In each replicate an adaptation phase of 15 days preceded a 5 days of total feces collection for digestibility trial and 1 day collection of fresh fecal samples for bacterial enumeration, pH measurement and determination of short chain fatty-acids and bioactive amines concentration. On day 21, a blood samples were collected for immunophenotypic quantification of lymphocyte subsets CD5+CD4+, CD5+, CD5+CD8+, CD45+ e CD45+CD21+ through flow cytometry. The data were evaluated using Proc GLM of SAS, means were compared by Tukey test and polinomial and orthogonal contrasts (p <0,1). The inclusion of YCW did not change the coefficient of total tract apparent digestibility of nutrients (p>0.1) nor the fecal bacterial enumeration (total aerobes and anaerobes, Bifidobacterium, Lactobacillus, Clostridium and Escherichia coli) in dog?s feces (p>0.1). Fecal butirate concentration increased linearly with YCW addition (p=0,055), fecal concentrations of tyramine (P=0,1) and histamine (P=0,07) decreased linearly, and phenylethylamine (P=0,07) and tryptamine (P=0,07) concentrations showed a quadratic reduction with YCW consumption. Dogs also presented a linear increase in lymphocytes CD5+ concentration (P=0,10) and a greater CD45+CD21+ lymphocytes number (P=0,053) with YCW addition. In conclusion YCW presents a prebiotic (changing bacterial end product formation) and immunostimulating effect for dogs

Antimicrobial activity of Lactobacillus and Bifidobacterium strains against pathogenic microorganisms in vitro / Atividade antimicrobiana de Lactobacillus e Bifodobacterium frente a microrganismos patogênicos in vitro

Lactobacilli and bifidobacteria have a long history of safe use in foods. These bacteria have biotechnological characteristics of interest such as the inhibition of pathogens. In this work, two lactobacilli strain and a bifidobacterium strain isolated from human gut were evaluated concerning to their ability to inhibit pathogenic microorganisms in foods by diffusion agar tests. Moreover, we assessed the metabolites produced in culture broth under static and shaking growth to simulate anaerobiosis and aerobiosis conditions, respectively. L. acidophilus LA5, L. plantarum DCTA 8420 and B. lactis DCTA 8724 showed ability to inhibit S. aureus FRI 196, strains producer toxins A and D, as well as B. cereus ATCC 25923, E. coli ATCC 25922 and S. Enteritidis, whose inhibition halos reached, on average, 24 mm in diameter. In the agar diffusion method with concentrated culture medium, it was possible to observe the effect of oxygen on the production of toxic substances. This result showed that cultivation of Lactobacillus under aerobic conditions seems to exert greater inhibitory effect, whereas for Bifidobacterium strain the effect was the opposite.

Lactobacilos e bifidobactérias apresentam um longo histórico de uso seguro em alimentos, além de apresentarem características de interesse biotecnológico como a inibição de patógenos. Neste trabalho duas linhagens de lactobacilos e uma de bifidobactéria, isoladas do intestino humano, foram avaliadas em testes de difusão em ágar, quanto à capacidade de inibição de microrganismos patogênicos de ocorrência comuns em toxinfecções alimentares. Adicionalmente, foram avaliados os metabólitos produzidos em caldo de cultivo estático e em agitação para simular condições de anaerobiose a aerobiose, respectivamente. As três bactérias, L. acidophilus LA5, L. plantarum DCTA 8420 e B. lactis DCTA 8724 apresentaram capacidade de inibição para S. aureus FRI 196 linhagem produtora de toxinas A e D, assim como para B. cereus ATCC 25923, E. coli ATCC 25922 e S. Enteritidis, cujos halos de inibição apresentaram em média 24 mm de diâmetro. No método de difusão em ágar com meio de cultivo concentrado, foi possível observar o efeito do oxigênio sobre a produção de substâncias tóxicas aos microrganismos indicadores, evidenciando que Lactobacillus cultivados em aerobiose, parece produzir metabólitos com maior efeito inibitório, enquanto que, para a linhagem de Bifidobacterium, o efeito foi oposto.

Antimicrobial activity of Lactobacillus and Bifidobacterium strains against pathogenic microorganisms in vitro / Atividade antimicrobiana de Lactobacillus e Bifodobacterium frente a microrganismos patogênicos in vitro

Lactobacilli and bifidobacteria have a long history of safe use in foods. These bacteria have biotechnological characteristics of interest such as the inhibition of pathogens. In this work, two lactobacilli strain and a bifidobacterium strain isolated from human gut were evaluated concerning to their ability to inhibit pathogenic microorganisms in foods by diffusion agar tests. Moreover, we assessed the metabolites produced in culture broth under static and shaking growth to simulate anaerobiosis and aerobiosis conditions, respectively. L. acidophilus LA5, L. plantarum DCTA 8420 and B. lactis DCTA 8724 showed ability to inhibit S. aureus FRI 196, strains producer toxins A and D, as well as B. cereus ATCC 25923, E. coli ATCC 25922 and S. Enteritidis, whose inhibition halos reached, on average, 24 mm in diameter. In the agar diffusion method with concentrated culture medium, it was possible to observe the effect of oxygen on the production of toxic substances. This result showed that cultivation of Lactobacillus under aerobic conditions seems to exert greater inhibitory effect, whereas for Bifidobacterium strain the effect was the opposite.

Lactobacilos e bifidobactérias apresentam um longo histórico de uso seguro em alimentos, além de apresentarem características de interesse biotecnológico como a inibição de patógenos. Neste trabalho duas linhagens de lactobacilos e uma de bifidobactéria, isoladas do intestino humano, foram avaliadas em testes de difusão em ágar, quanto à capacidade de inibição de microrganismos patogênicos de ocorrência comuns em toxinfecções alimentares. Adicionalmente, foram avaliados os metabólitos produzidos em caldo de cultivo estático e em agitação para simular condições de anaerobiose a aerobiose, respectivamente. As três bactérias, L. acidophilus LA5, L. plantarum DCTA 8420 e B. lactis DCTA 8724 apresentaram capacidade de inibição para S. aureus FRI 196 linhagem produtora de toxinas A e D, assim como para B. cereus ATCC 25923, E. coli ATCC 25922 e S. Enteritidis, cujos halos de inibição apresentaram em média 24 mm de diâmetro. No método de difusão em ágar com meio de cultivo concentrado, foi possível observar o efeito do oxigênio sobre a produção de substâncias tóxicas aos microrganismos indicadores, evidenciando que Lactobacillus cultivados em aerobiose, parece produzir metabólitos com maior efeito inibitório, enquanto que, para a linhagem de Bifidobacterium, o efeito foi oposto.