RESUMEN
El objetivo de este estudio es caracterizar molecularmente bacterias con potencial probiótico aisladas de heces de neonatos humanos. Se evaluó 60 muestras de heces de neonatos (0-3 días) se enriquecieron en caldo Man Rogosa y Sharp (MRS) a 37°C/24h. Se seleccionó y se sometió a pruebas in vitro con sales biliares, resistencia a pH bajo y actividad antimicrobiana frente a Escherichia coli ATCC25922, E. coli ATCC35218, Salmonella enterica y Listeria inocua mediante el ensayo difusión en agar. La identificación molecular se realizó con amplificaciones PCR-BOX y el secuenciamiento del gen 16S rRNA. Se aislaron un total de 48 cepas y todas presentaron resistencia a pH 3 y 0.3% sales biliares; 3 cepas mostraron actividad antimicrobiana frente a E. coli ATCC25922, 1 cepa frente a E. coli ATCC35218, 5 cepas frente a L. inocua y todas frente a Salmonella entérica. De las 48 cepas se obtuvieron dos perfiles BOX-PCR pertenecientes a los géneros de Lactobacillus y Enterococcus. Nueve cepas (C5(2), C6(1), C7(1), C11(2), C16 2, C19(2), C20, C35, y C42) presentaron un 100% de similaridad a Lactobacillus plantarum ATCC 14917T [ACGZ01000098] y dos cepas (C15 y C40) un 99.93% y 99.80% de similaridad, respectivamente a Enterococcus faecium CGMCC 1.2136T [AJKH01000109]; estas cepas mostraron actividad en leche con diferencias significativas (p valor < 0.05) en la cinética de pH 3. En conclusión se encontró bacterias con potencial probiótico.
The aim of this study is to molecularly characterize bacteria with probiotic potential isolated from feces of human neonates. Sixty stool samples from neonates (0-3 days) were evaluated and enriched in Man Rogosa and Sharp (MRS) broth at 37 ° C / 24h. It was selected and subjected to in vitro tests with bile salts, resistance to low pH and antimicrobial activity against Escherichia coli ATCC25922, E. coli ATCC35218, Salmonella enterica and Listeria inocua by agar diffusion assay. The molecular identification was made with PCR-BOX amplifications and the sequencing of the 16S rRNA gene. A total of 48 strains were isolated and all showed resistance to pH 3 and 0.3% bile salts; 3 strains showed antimicrobial activity against E. coli ATCC25922, 1 strain against E. coli ATCC35218, 5 strains against L. innocuous and all against S. enterica. Of the 48 strains, two BOX-PCR profiles belonging to the genera of Lactobacillus and Enterococcus were obtained. Nine strains (C52, C61, C71, C112, C162, C192, C20, C35, and C42) presented 100% similarity to L. plantarum ATCC 14917T [ACGZ01000098] and two strains (C15 and C40) 99.93% and 99.80 % similarity, respectively to Enterococcus faecium CGMCC 1.2136T [AJKH01000109]; these strains showed activity in milk with significant differences (p value <0.05) in the kinetics of pH 3. In conclusion, bacteria with probiotic potential were found.
RESUMEN
Se presenta un estudio etnobotaÌnico sobre las Leguminosas (Fabaceae) silvestres de uso medicinal de una regioÌn altoandina del PeruÌ (distrito de Lircay, provincia de Angaraes, departamento de Huancavelica). La metodologiÌa se basoÌ en entrevistas semiestructuradas a 33 informantes clave, calculando el iÌndice de Importancia Cultural. Se registran 12 geÌneros con 17 especies. Todas las plantas poseen por lo menos un nombre vernaÌculo y tienen usos adicionales a la categoriÌa medicinal. Dos especies son comercializadas en Lircay: Desmodium molliculum y Otholobium pubescens. Se reportan nueve categoriÌas de uso, siendo la categoriÌa medicinal la que presentoÌ mayor nuÌmero de especies (17 especies) y la que tuvo maÌs reportes de uso (125 reportes). La Leguminosa con mayor Importancia Cultural es Senna versicolor, lo que demuestra que es la especie maÌs arraigada al conocimiento tradicional de los pobladores locales.
What follows is an ethnobotanical study of wild medicinal Legumes from a high Andean region of Peru (district of Lircay, province of Angaraes, department of Huancavelica). The methodology was based on 33 semi-structured interviews with key informants, and the Cultural Importance Index was calculated. A total of 12 genera with 17 species were recorded. All of the plants possessed at least one vernacular name and in addition to their medicinal function had other uses. Two species are traded in the Lircay: Desmodium molliculum and Otholobium pubescens. Nine categories of use were reported, with the medicinal category registering the highest number of species (17 species) and the greatest number of reported uses (125 reports). The legume of greatest cultural importance is Senna versicolor, which was shown to be the most deeply-rooted plant in the traditional knowledge of the local.