Short-chain fatty acids production by Bifidobacterium species in the presence of salep

BACKGROUND: Salep is obtained by grinding dried orchid tubers and used as a valuable ingredient in the food industry. Because of the glucomannan content of salep, it is thought to have prebiotic potential. However, there is little information in studies concerning the fermentation characteristics and potential prebiotic properties of salep. The objective of this study was to investigate the effect of salep on bifidobacterial growth by measuring the highest optical density (OD), calculating the specific growth rates, and determining the production of lactic acid and short-chain fatty acids (acetic, propionic, and butyric acid) as a result of bacterial fermentation. RESULT: The OD and pH values obtained in this study showed that salep was utilized as a source of assimilable carbon and energy by the Bifidobacterium species (BS). All Bifidobacterium strains produced lactic, acetic, propionic, and butyric acid, indicating that salep is readily fermented by these bacteria. Salep at 1% (w/v) showed a similar effect on bifidobacterial growth as that promoted by 1% (w/v) glucose used as a traditional carbon source. CONCLUSIONS: Bifidobacterium species can develop in media containing salep as well as in glucose and exhibit the potential to be used as new sources of prebiotics.

Effects of probiotic intake on intestinal bifidobacteria of celiac patients / Efeitos do consumo de probióticos sobre as bifidobactérias intestinais de pacientes celíacos

ABSTRACT BACKGROUND Healthy individuals exhibit a significantly higher concentration of faecal bifidobacteria in comparison to celiac patients. Even though there are potential benefits in probiotic usage, they have been little explored as an adjunctive therapy in celiac disease. OBJECTIVE This study aimed at the comparison of faecal bifidobacteria concentration and pH among celiac patients and healthy subjects before and after the daily intake of 100 g of yogurt containing probiotic for a thirty-day period. METHODS Feces from 17 healthy subjects and 14 celiac patients were analyzed, in which stool culture was performed for the isolation and quantification of faecal bifidobacteria. Furthermore, Gram’s method was employed for the microscopic analysis of the colonies, while the identification of the Bifidobacterium genus was made through determination of the fructose-6-phosphate phosphoketolase enzyme. Faecal pH was measured using a calibrated pHmeter. RESULTS Faecal bifidobacteria concentration before probiotic consumption was significantly higher in healthy individuals (2.3x108±6.3x107 CFU/g) when compared to celiac patients (1.0x107±1.7x107 CFU/g). Faecal pH values did not show a significant difference. After the daily consumption of probiotic-containing yogurt both groups showed a significant increase in the concentration of faecal bifidobacteria, but healthy subjects presented significantly higher bifidobacteria concentrations (14.7x108±0.2x108 CFU/g) than the celiac group (0.76x108±0.1x108 CFU/g). The obtained pH values from both groups were not significantly different, being 7.28±0.518 for the celiac patients and 7.07±0.570 for healthy individuals after the probiotic intake. CONCLUSION The probiotic supplementation significantly increased the number of bifidobacteria in the feces of celiac patients, although it was not sufficient to reach the concentration found in healthy individuals prior to its consumption.

RESUMO CONTEXTO Indivíduos saudáveis apresentam uma concentração de bifidobactérias fecais significativamente maior em comparação a pacientes celíacos. Apesar de haver benefícios potenciais no uso de probióticos na doença celíaca, estes têm sido pouco explorados como uma terapia adjuvante. OBJETIVO Este estudo objetivou a comparação do pH e concentração fecal de bifidobactérias entre pacientes celíacos e indivíduos saudáveis antes e após o consumo diário de 100 g de iogurte contendo probiótico por um período de 30 dias. MÉTODOS Foram analisadas fezes de 17 pessoas saudáveis e 14 pacientes celíacos, tendo sido realizada a coprocultura para o isolamento e quantificação de bifidobactérias fecais. Além disso, o método de Gram foi empregado na análise microscópica das colônias, enquanto a identificação do gênero Bifidobacterium foi feita através da determinação da enzima frutose-6-fosfato fosfocetolase. O pH fecal foi medido usando um pHmetro calibrado. RESULTADOS A concentração de bifidobactérias fecais antes do consumo do iogurte probiótico foi significativamente maior em indivíduos saudáveis (2.3x108±6.3x107 UFC/g) quando comparada aos celíacos (1.0x107±1.7x107 CFU/g). Por outro lado, o pH fecal de ambos os grupos não apresentou diferença significativa. Após o consumo diário de iogurte contendo probiótico, ambos os grupos tiveram um aumento significativo na concentração de bifidobactérias fecais, entretanto indivíduos saudáveis apresentaram concentrações de bifidobactérias significativamente maiores (14.7x108±0.2x108 UFC/g) do que o grupo celíaco (0.76x108±0.1x108 UFC/g). Os valores de pH obtidos de ambos os grupos não foram significativamente diferentes, sendo de 7.28±0.518 para os pacientes celíacos e de 7.07±0.570 para os indivíduos saudáveis após o consumo do probiótico. CONCLUSÃO A suplementação com probiótico aumentou significativamente o número de bifidobactérias nas fezes dos pacientes celíacos apesar de não ter sido suficiente para alcançar a concentração encontrada em indivíduos saudáveis antes do consumo de probióticos.

Lower bifidobaceeria counts in adult patients with celiac disease on a gluten-free deet / Baixa contagem de bifidobactérias em pacientes adultos com doença celíaca, em dieta isenta de glúten

Context The ingestion of gluten is responsible for the symptoms of Celiac disease, but other environmental factors can also influence. Strains of the Bifidobacterium genus have been shown to afford protection against the inflammatory response and mucosal damage caused by gliadin peptides in vitro. Objectives This study was designed to compare the concentration of fecal bifidobacteria and pH of patients with celiac disease on gluten-free diet and control subjects in order to identify if the imbalance on fecal microbiota still remain during the treatment of celiac disease and identify the necessity of dietary supplementation with pre- or probiotics. Methods It was analyzed the feces of 42 healthy subjects and 14 celiac patients. The bifidobacteria count in feces was done in selective medium BIM-25. Microscopic analysis of the colonies was performed by Gram stain. The identification of the genus Bifidobacterium was performed by determination of fructose-6-phosphate phosphoketolase. Fecal pH was measured using a pH meter. Results The concentration of bifidobacteria per gram of feces was significantly higher in healthy subjects (controls) (1.5 ± 0.63 x108 CFU/g) when compared to celiac patients (2.5 ± 1.5 x107 CFU/g). The fecal pH was not different between celiac patients (7.19 ± 0.521) and controls (7.18 ± 0.522). Conclusions These results suggest that with lower levels of bifidobacteria, celiac patients have an imbalance in the intestinal microbiota, regardless of pH, even while on a gluten-free diet. This fact could favor the pathological process of the disorder. .

Contexto A ingestão do glúten é responsável pelos sintomas da doença celíaca, mas outros fatores ambientais também podem influenciar. Tem sido mostrado que as cepas do género Bifidobacterium proporcionam proteção contra a resposta inflamatória, lesão da mucosa causada por péptidos da gliadina in vitro. Objetivos Este estudo foi desenvolvido para comparar as concentrações de bifidobactérias e pH fecal de pacientes com doença celíaca em dieta isenta de glúten e indivíduos controles, a fim de identificar se o desequilíbrio na microbiota fecal ainda permanece durante o tratamento da doença celíaca e, identificar a necessidade de suplementação alimentar com pré ou probióticos. Métodos Foram analisadas as fezes de 42 indivíduos saudáveis e 14 pacientes com doença celíaca. A contagem de bifidobactérias nas fezes foi feita em meio seletivo BIM-25. A análise microscópica das colônias foi realizada por coloração de Gram. A identificação do género Bifidobacterium foi realizada por determinação de phosphoketolase frutose-6-fosfato. O pH fecal foi medido usando um medidor de pH. Resultados As concentrações de bifidobactérias por grama de fezes foi significativamente mais elevada nos indivíduos saudáveis (controles) (1,5 ± 0,63 x108 UFC/g), quando comparada com pacientes com doença celíaca (2,5 ± 1,5 x107 UFC/g). O pH fecal não foi diferente entre pacientes celíacos (7,19 ± 0,521) e controles (7,18 ± 0,522). Conclusões Estes resultados sugerem que, com concentrações inferiores de bifidobactérias, pacientes com doença celíaca tem um desequilíbrio na microbiota intestinal, independentemente do pH, mesmo durante uma dieta isenta de ...

Kinetic analysis and effect of culture medium and coating materials during free and immobilized cell cultures of Bifidobacterium animalis subsp. lactis Bb 12

Microencapsulation technique appears helpful for more protection of Bifidobacteria against acid inhibitory effect. The effect of medium composition and product inhibitory in free cell culture, as well as the effect of the coating materials in immobilized cells, on biomass growth, acid production and substrate utilization kinetics of Bifidobacterium animalis subsp. lactis Bb 12 in uncontrolled batch fermentation was examined. The Monod and the Luedeking and Piret equations with a product inhibition term involving toxic power terms improved model efficiency for both growth and production. The model showed that media and coating materials had an effect on toxic power terms. Cell immobilization had a positive impact on B. animalis culture. Kinetic analysis revealed the permeability of the coating material had a major impact on culture parameters; permeability increased in the following way: Gellan xanthan < Alginate chitosan < K-Carageenan-locust been, and hence growth parameters x m, maximum specific growth rate (h-1) (um) and monod constant (g lactose L-1) (K S) followed the same trend as well as the linking between growth and production. The link between the microbial environment and cell growth was highlighted by the model. It was shown that for an increasing protect effect of coating materials against environmental deleterious factors, namely a decrease of the permeability, transport limitation occurred, which was disadvantageous for cell formation.

Regulación de la microbiota intestinal en voluntarios sanos mediante el consumo de un producto con el probiotico Lactobacillus johnsonii La1 / Regulation of the intestinal microbiota of healthy volunteers through the intake of the probiotic lactobacillus johnsonii la1

A basic mechanism implicated in the human health-promoting properties of probiotics is their ability to maintain the homeostasis of the intestinal microbiota. This study evaluates how the ingestion of different amounts of the probiotic Lactobacillus johnsonii La1 (La1) influences the main bacterial populations of the fecal microbiota. Eight asymptomatic volunteers participated in the study. After a basal period, they ingested 100 mL of a product containing 10(8) La1/mL during the first week, 200 mL during the second week and 500 ml during the third week. Fecal samples were obtained at the end of each period and during the 2 weeks post-ingestion. Lactobacilli were determined by culture on MRS agar and La1 colonies were confirmed by ERIC-PCR. The main populations of fecal bacteria were identified by fluorescent probes and flow cytometry. At baseline, 18.3 percent of the total fluorescent bacteria were F. praunitzii, 13.2 percent Bacteroides, 2.05 percent Bifidobacterium and 0.95 percent Lactobacillus. Fecal excretion of La1 increased during the ingestion period but it was cleared from the stools of the volunteers 2 weeks later. La1 intake increased the populations of Lactobacillus (p=0.056) and Bifidobacterium (p=0.067), which are considered as beneficial for the host, while it decreased those of F. prausnitzii (p=0.005) a potentially pathogenic microorganism. These bacterial populations returned to their baseline levels during the post-ingestion period. The regular intake of a La1-containing product beneficially affects the homeostasis of the human fecal microbiota probably contributing to the health-promoting effects of this probiotic.

Uno de los principales mecanismos implicados en las propiedades saludables de los probióticos es su capacidad de mantener la homeostasis de la microbiota intestinal. Este estudio evalúa cómo el consumo de distintas cantidades del probiótico Lactobacillus johnsonii La1 (La1) contribuye en la modulación de las principales poblaciones de la microbiota fecal. Ocho voluntarios asintomáticos participaron en el estudio. Después de un periodo basal, consumieron 100 mL de un producto que contenía 10(8) La1/mL durante la primera semana, 200 mL durante la segunda semana y 500 mL durante la tercera semana. Se obtuvieron muestras de deposición al final de cada uno de estos períodos y luego a los 7 y 14 días de haber terminado el consumo del producto. Las cantidades de lactobacilos excretados fueron determinadas por cultivo en agar MRS y las colonias de La1 fueron confirmadas por ERIC-PCR. Algunas de las principales poblaciones de bacterias fecales fueron evaluadas por hibridación in situ con sondas fluorescentes (FISH) y citometría de flujo. A nivel basal, 18.3 por ciento del número total de bacterias fluorescentes detectadas eran F. praunitzii,13.2 por ciento Bacteroides, 2.05 por ciento Bifidobacterium y 0.95 por ciento Lactobacillus. La excreción fecal de La1 aumentó durante el período de consumo pero desapareció después de 14 días de haber terminado el período de ingestión. El consumo de La1 aumentó las poblaciones de Lactobacillus (p=0.056) y Bifidobacterium (p=0.067) que son consideradas como beneficiosas para el huésped mientras que disminuyó aquella de F. prausnitzii (p=0.005), un microorganismo potencialmente patogénico. Las poblaciones bacterianas afectadas volvieron a sus niveles basales durante el periodo post-ingestión. Estos resultados indican que el consumo regular de La1 modula la homeostasis de la microbiota intestinal, lo cual contribuye probablemente a los efectos beneficiosos de este probiótico sobre la salud.

Evaluación de la actividad de cultivos probióticos sobre Listeria monocytogenes durante la producción y almacenamiento de yogur / Evaluation of the activity of probiotic cultures over Listeria monocytogenes during the production and storage of yogurt

The effect of probiotic cultures over Listeria monocytogenes during the production and storage of yogurt was evaluated. A yogurt mixture (10.6 per cent non-fat solid liquids, 3 per cent fat and 0.3 per cent gelatin) was prepared, homogenized and pasteurized. Yogurt was inoculated with 0, 10(2), 10(4) and 10(6) CFU/mL of L. monocytogenes and 0.02 per cent of traditional lactic culture YC 180 (Streptococcus thermophilus and Lactobacillus bulgaricus) and probiotic culture ABY-1 (Bifidobacterium longum, B. bifidum, B, infantis, Lactobacillus acidophilus, Streptococcus thermophilus y Lactobacillus delbrueckii subsp. bulgaricus). It was incubated for 3 h at 43 degrees C until pH reached an approximate value of 4.8, followed by refrigeration at 5 degrees C for 21 days. During fermentation, samples were taken every hour, and during storage every 3 days, analyzing pH and lactic, bifidobacteria and pathogen count for each time. It was demonstrated that there was no significant simple effect for the type of culture used (ABY-1 and YC 180) (p = 0.684) over the amount of L. monocytogenes present in yogurt during the fermentation and storage periods. The presence of bifidobacteria in the ABY-1 culture did not present a significant effect over L. monocytogenes. Neither the effect of time presented a significant effect over L. monocytogenes (p = 0.448). In this case, the ABY-1 and YC 180 cultures present a bacteriostatic effect over the pathogen. The probiotic cultures had a bacteriostatic but not bactericidal effect over L. monocytogenes. This is not related to the protective effect of these cultures in bowel, since in-vivo conditions favor the production of antimicrobial substances, such as bacteriocins that act over pathogens.

Importância dos oligossacárides (prebióticos) como promotores de crescimento de bifidobactéria / Importance of oligossacharids as stimulate Bifidobacteria growth

Nos últimos anos têm-se intensificado as pesquisas em relaçäo aos oligossacárides, por serem carboidratos que estimulam o crescimento das bifidobactérias no intestino. Esses microrganismos säo benéficos à saúde humana e por isso a sua permanência na microbiota intestinal é desejável. Os oligossacárides produzidos em maior quantidade säo os frutoligossacárides, lactulose, galactoligossacárides, isomaltoligossacárides e maltoligossacárides. Säo empregados na elaboraçäo de alguns alimentos como bebidas, leite em pó, produtos de confeitaria e sobremesas lácteas, sendo a maioria destes produtos encontrados no Japäo. Os oligossacárides também podem ter outras aplicaçöes, como produçäo de cosméticos, alimentos dietéticos, produtos anticariogênicos e alimentaçäo de animais. Entretanto, ainda säo necessárias mais pesquisas que garantam o seu efeito benéfico e a segurança ao hospedeiro.

Un método sencillo para generar anaerobiosis en tubos de cultivo / A simple method to achieve anaerobiosis in culture media tubes

Introducción. El cultivo de bacterias anaerobias requiere una atmósfera libre de oxígeno, por lo cual, a pesar de la gran importancia clínica de los anaerobios, el diagnóstico usualmente es sólo microscópico. En este estudio se propone y evalúa un método para preparar tubos de medio de cultivo libres de oxígeno, sencillo, económico y al alcance de cualquier laboratorio. Material y métodos. La atmósfera anaerobia se logra durante el proceso de autoclavado y descompresión rápida de los tubos a los que se le ha incorporado un tapón de hule, tapa de rosca y una aguja a través de la cual se elimina la atmósfera aerobia. Para evaluar la efectividad de estos medios se crecieron 45 especies de clostridium y 12 especies de los géneros Bifidobacterium, acteroides, Lactobacillus, Propionibacterium y Actinomyces, (hasta completar 80 cepas), en tubos anaeróbios prerreducidos antes de esterilizar (PRAS) y en tubos preparados de acuerdo con el método propuesto. Resultados. Las 80 cepas de bacterias anaerobias crecieron en los tubos prerreducidos y sin prerreducir, no hubo diferencias en los grados de turbiedad. Discusión. El método propuesto para generar anaerobiosis en los tubos de cultivo es una posibilidad al alcance a cualquier laboratorio de bacteriología, para mejorar el diagnóstico, mediante cultivo, de infecciones por bacterias anaerobias