Bifidobacterium scardovii aislado de 2 muestras consecutivas de orina / Bifidobacterium scardovii isolated from 2 consecutive urine samples

La especie Bifidobacterium scardovii está constituida por bacilos gram positivos anaerobios facultativos, cuyo desarrollo es estimulado por el CO2 y la anaerobiosis. Excepcionalmente se la ha asociado a infecciones humanas. Se presenta el caso de un paciente anoso con infección urinaria por B. scardovii y Enterococcus faecalis, ambos microorganismos aislados en 2 urocultivos consecutivos. El bacilo no desarrolló en los medios de cultivo habituales, pero sí en agar chocolate en CO2 y en agar Brucella suplementado, incubados durante 72 h a 35°C. La coloración de Gram alertó acerca de su presencia al observarse abundantes bacilos gram positivos irregulares con extremos bifurcados en forma de Y, y escasos cocos gram positivos. Es importante la coloración de Gram en orinas con piuria y la siembra en medios enriquecidos por tiempos prolongados. En este caso, sin el resultado del Gram y con el desarrollo de E. faecalis, no hubiésemos advertido la presencia del agente mayoritario.

Bifidobacterium scardovii species consists of facultative anaerobic gram-positive rods whose growth is stimulated by CO2 and anaerobiosis. Exceptionally it has been associated with infections in humans. An elderly male patient with a urinary tract infection due to B. scardovii and Enterococcus faecalis is presented here; both microorganisms were isolated from two consecutive urine samples. The bacillus did not grow on standard media, but on chocolate agar incubated in CO2 and on supplemented Brucella agar in an anaerobic atmosphere, incubated for 72 h at 35°C. Gram staining with abundant irregular gram-positive rods with Y-shaped ends and some gram-positive cocci alerted to its presence. The importance of the Gram stain test in urine samples with pyuria and the growth on enriched media for long periods is highlighted here. In this case, if we had not had the Gram stain test results, and had considered only the E. faecalis growth, we would have lost the major etiologic agent.

Efeito do leite probiótico fermentado na resposta imune celular em cólon de camundongos BALB/c / Effect of probiotic fermented milk in immune cellular response of BALB/c mice colon

O principal crescimento na indústria de alimentos funcionais corresponde ao dos produtos probióticos e prebióticos. A literatura mostra efeitos imunomoduladores de certas cepas probióticas, contudo, os resultados são às vezes controversos e os mecanismos implicados ainda são pouco elucidados. Sabe-se, no entanto que algumas cepas de probióticos aumentam significantemente a liberação de IL-10 e γ-INF modulando a resposta imune, além destas respostas serem de forma mais branda relacionada às bactérias Gram-positivas probióticas do que às Gram-positivas patogênicas. O presente trabalho teve como objetivo geral estudar o efeito do leite probiótico fermentado na resposta imune celular em cólon de camundongos BALB/c. Os objetivos específicos foram: (i) determinar o efeito imunomodulador do leite adicionado de probiótico em camundongos normais, (ii) identificar os tipos celulares implicados na resposta imune específica por citometria de fluxo e, (iii) colocalizá-los nos cortes histológicos. Simultaneamente, a análise e a comparação da resistência do probiótico à digestão gastrintestinal in vitro e a produção de metabólitos bioativos de acordo com os deferentes produtos foi realizada. Foram preparados leites nos quais as variáveis estudadas foram a tecnologia empregada para a produção das formulações (a) leite; (b) água, (c) leite não fermentado; (d) leite fermentado; (e) leite fermentado seguido de pasteurização, usando a mesma concentração da cepa comercial Bifidobacterium animalis subsp. lactis HOWARU HN019. O leite desnatado e a água foram usados como controles

Functional food industry is in expansion mainly due to probiotic and prebiotic products. Studies have shown some probiotic strains develop immune modulation effect, however, these results are controversial and the mechanisms are not been well understood. Although, some probiotic strains increase IL-10 and γ-INF release modulating immune response, this response is weaker in probiotic strains when compared to pathogenic Gram-positive bacteria. The major aim of the present study was to assess the effect of probiotic fermented milk in cellular immune response of Balb/c mice colon. The specific objectives were: (i) to determine the immunomodulation of the milk added of probiotic in normal mice; (ii) to identify the cellular types implied in immune specific response and, (iii) to colocalize them in histological sections. Besides, the analyze and comparation of the probiotic resistance upon in vitro gastrointestinal and bioactive metabolites release in fermented or unfermented bifido milk using the same matrix, probiotic strain and probiotic dose in CFU. mL-1 were conducted. Dairy products were prepared in which variable form of technological appliance were: (i) milk, (ii) water, (iii) unfermented milk, (iv) fermented milk, and (v) fermented and heat treatment milk, all using Bifidobacterium subsp. lactis HOWARU HN019 strain in the same concentration. The skimmed milk and water were used as controls. The immune effects were evaluated by histological sections and the lymphocytic infiltrated was analyzed by flow citometry and histology

Capacidad bactericida de Bifidobacterium sp. aislada de leche materna y de heces de neonatos, frente a los principales causantes de enfermedades transmitidas por alimentos / Antibiotic activity of Bifidobacterium sp. isolated from breast milk and newborn faeces, against the main causes for foodborne illnesses

Introducción. La microbiota del tubo digestivo humano contiene bacterias benéficas para la salud que regulan el funcionamiento del colon e inhiben algunos microorganismos patógenos intestinales. Las bifidobacterias aisladas de neonatos y de leche materna se usan como microorganismos probióticos para prevenir enfermedades infecciosas, incluidas las transmitidas por alimentos. Objetivo. Aislar e identificar Bifidobacterium sp. en humanos y determinar su capacidad bactericida frente a patógenos causantes de enfermedades transmitidas por alimentos, importantes en Colombia y en el mundo. Materiales y métodos. Se recolectaron 17 muestras de leche materna, y 19 muestras de meconio y heces de neonatos, en diferentes hospitales de Bogotá. Los 26 aislamientos sospechosos se identificaron a nivel de género mediante PCR 16-23S; para la identificación de especie, se secuenciaron algunos de los aislamientos. La capacidad antagonista de las 26 cepas de Bifidobacterium sp. fue evaluada contra Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Salmonella enteritidis ATCC 13076, Listeria monocytogenes ATCC 7644 y E. coli O157:H7 ATCC 35150. Para las cepas que presentaron mayor actividad antagonista, se analizó el extracto inhibidor con ensayos de difusión en placa. Resultados. Todas las cepas amplificaron la banda esperada para la confirmación de género; asimismo, las cepas Bif 013 y Bif 023 se identificaron por secuenciación como Bifidobacterium breve, con una homología del 97%. Del total de cepas, 17 mostraron capacidad de inhibir, al menos, uno de los patógenos evaluados. E. coli ATCC 25922 fue el patógeno más inhibido. Se determinó que la cepa Bif 023 es eficiente como antagonista, ya que inhibió todos los patógenos evaluados. Los halos de inhibición presentaron diámetros mayores a lo esperado, lo que indica una muy buena capacidad antagonista de las cepas nativas. Se aisló Bifidobacterium sp. de leche materna, meconio y heces de neonatos, por lo cual se confirmó que este microorganismo es microbiota humana (2). Conclusión. Se concluye que, debido a la gran capacidad antagonista de la mayoría de las bacterias aisladas, éstas pueden estar cumpliendo una importante función protectora en el recién nacido, en particular las cepas Bif 013 y Bif 023 aisladas de materia fecal. Estos microorganismos deben continuar siendo estudiados para definir su potencial probiótico. También, se pueden evaluar para bioconservación en la industria y contra patógenos transmitidos por alimentos.

Introduction. The microbiota in the human gastrointestinal tract contains beneficial microorganisms for human health, which contribute to the regulation of colonic function and inhibition of some intestinal pathogens growth. Bifidobacterium sp. isolated from newborns and breast milk are used as probiotic microorganisms, which are useful in the prevention of infectious diseases including foodborne illnesses. Objective: To isolate and identify human Bifidobacterium sp. and to determine its antibiotic activity against important pathogens which cause foodborne illnesses in Colombia and the world. Materials and methods. 17 breast milk samples and 19 meconium and newborn faeces samples were collected from different hospitals in Bogotá. 26 presumptive Bifidobacterium strains were identified at genus level by PCR 16-23S; some strains were identified at species level by nucleic acid sequencing. The antagonistic activity of 26 Bifidobacterium sp. strains was tested against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Salmonella enteritidis ATCC 13076, Listeria monocytogenes ATCC 7644 and E. coli O157:H7 ATCC 35150. For the strains that showed a greater antibiotic activity, the inhibitory compound was analyzed using disk diffusion tests. Results. All strains amplified the expected band for genus confirmation. Strains Bif 023 and Bif 013 were identified by DNA-sequencing as Bifidobacterium breve, with 97% homology. 17 strains were able to inhibit at least one of the pathogens tested. Escherichia coli ATCC 25922 was the most inhibited. It was determined that the strain Bif 023 is highly efficient as an antagonist strain due its ability to inhibit all the evaluated pathogens. Inhibition areas showed higher diameters than expected, suggesting an enhanced antagonist capacity of native strains. Bifidobacterium sp. was isolated from breast milk, meconium and newborn faeces which confirmed that this microorganism is human microbiota. Conclusion. Due to the high antagonist activity of most isolated bacteria, they could be playing an important protective function in the newborn, in particular strains Bif 013 and Bif 023, isolated from faeces. Other studies must be performed with these organisms to determine their probiotic potential as well as their use in the biocontrol industry due their activity against foodborne pathogens.

Influence of probiotics on Candida presence and IgA anti-Candida in the oral cavity

Probiotics are defined as microorganisms that promote benefits to host health, mainly by regulating resident microbiota. Disequilibrium in microbiota can favor the growth of opportunist microorganisms and the development of pathologies, like candidosis caused by yeasts of the Candida genus. This work evaluated whether probiotics consumption was able to influence a specific immunological response to Candida and the presence of these yeasts in the oral cavity. Saliva samples were collected from healthy individuals and plated in Dextrose Saboraud Agar with chloramphenicol. Individuals presenting Candida in the oral cavity used the probiotic Yakult LBâ for 20 days, after which new collections and identifications were performed. Anti-Candida IgA analysis was conducted using the ELISA technique. Analysis of the results showed a significant reduction in Candida prevalence (46 percent) and mean Candida CFU/mL counts (65 percent). The Candida species identified were C. albicans (98 percent) and C.tropicalis (2 percent), before and after probiotics consumption. Immunological analysis demonstrated a significant reduction in anti-Candida IgA levels after probiotics use, probably due to less antigenic stimulation. In conclusion, in the individuals studied, probiotics use significantly reduced the amount of Candida in the oral cavity, possibly due to competition between the yeasts rather than by specific secretory immune response stimulation.

Evaluation of culture media for counts of Bifidobacterium animalis subsp. lactis Bb 12in yoghurt after refrigerated storage / Avaliação de meios de cultura para contagem de Bifidobacterium animalis subsp. lactis Bb 12em iogurte após a estocagem refrigerada

The agar RCPB pH5 has been considered a good alternative for counts of Bifidobacterium in yoghurt. However, during the refrigerated storage of yoghurt it is extremely difficult to count this microorganism due to the size of the colonies, which are so small they require the aid of a stereoscope to count them. Another agar, MRS-LP, has been also recommended for counts of Bifidobacterium in the presence of yoghurt bacteria. This study evaluated the supplementation of RCPB pH5 agar with dehydrated liver extract and the salts KH2PO4, K2HPO4, FeSO(4)7H2O, MnSO4H2O and MgSO(4)7H2O, aiming at improving the differentiation of Bifidobacterium in yoghurt after refrigerated storage, and also evaluated the selective count of Bifidobacterium in yoghurt using the agar MRS-LP. The agar MRS-LP presented the same cell recovery as non-fortified RCPB pH5 agar, used as a standard medium, thus being considered a good option for counts of Bifidobacterium in yoghurt. The fortified RCPB pH5 also presented the same recovery as the standard RCPB pH5 medium, however, the addition of dehydrated liver extract to the RCPB pH5 agar considerably increased the size of the Bifidobacterium colonies after refrigerated storage, making differentiation of the colonies much easier and reliable when compared to the standard non-fortified RPCP pH5. The addition of the salts (KH2PO4, K2HPO4, FeSO(4)7H2O, MnSO4H2O and MgSO(4)7H2O) had no influence on the performance of the RCPB pH5 agar.

O meio RCPB pH5 tem sido considerado uma boa opção para a contagem de Bifidobacterium em iogurte. Entretanto, durante a estocagem refrigerada do iogurte é extremante difícil a contagem deste microrganismo devido ao pequeno diâmetro desenvolvido pelas colônias de Bifidobacterium neste meio, sendo que a sua contagem somente se torna possível com o auxílio de um estereoscópio. Outro meio, MRS-LP, também tem sido recomendado para a contagem de Bifidobaterium em iogurte. Este estudo avaliou a suplementação do meio RCPB pH5 com extrato de fígado desidratado e com os sais KH2PO4, K2HPO4, FeSO(4)7H2O, MnSO4H2O e MgSO(4)7H2O, visando melhorar a diferenciação de Bifidobacterium em iogurte durante a estocagem refrigerada e também avaliou a contagem seletiva de Bifidobacterium em iogurte usando o meio MRS-LP. O meio MRS-LP apresentou a mesma recuperação de células que o meio RCPB pH5, usado como padrão, após 30 dias de estocagem refrigerada do iogurte, sendo considerado uma boa opção para a contagem de Bifidobacterium em iogurtes durante a estocagem refrigerada. O meio RCPB pH5 fortificado também apresentou a mesma recuperação de células de Bifidobacterium que o meio padrão RCPB pH5; entretanto, a adição de extrato de fígado desidratado aumentou consideravelmente o diâmetro das colônias de Bifidobacterium, tornando a diferenciação destas bastante fácil e confiável quando comparadas à sua diferenciação no meio RCPB pH5 sem a fortificação. A adição dos sais (KH2PO4, K2HPO4, FeSO(4)7H2O, MnSO4H2O e MgSO(4)7H2O) não exerceu influência no desempenho do meio RCPB pH5.