Aislamiento e identificación de Lactobacillus spp. de chicha de jora y caracterización del potencial probiótico in vitro
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Date
2024
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Universidad Nacional de Trujillo
Abstract
El presente trabajo tuvo como propósito aislar Lactobacillus spp. de chicha de jora artesanal provenientes de tres establecimientos productores del centro minorista “La Recoleta” del departamento de Cajamarca. Se aislaron 31 cultivos de BAL de los cuales solo ocho cumplieron con las características morfológicas, tintoriales y bioquímicas correspondientes a Lactobacillus spp. Se les sometió a pruebas de resistencia al pH ácido, tolerancia a las sales biliares, actividad hemolítica, capacidad de licuefacción de la gelatina, susceptibilidad a los antibióticos y actividad antimicrobiana para evaluar para evaluar su potencial probiótico. Los resultados obtenidos mostraron que a pH 4 todos los cultivos presentaron supervivencia igual o mayor al 45 %, solo 6 presentaron resistencia cuando fueron sometidos a pH 3 y ningún cultivo presentó resistencia a pH 2. Cuando fueron sometidos a concentraciones de 0.3% y 0.5% de sales biliares todos los cultivos presentaron una supervivencia mayor al 90 %. En las pruebas de virulencia ninguno cultivo presentó actividad proteolítica y solo un cultivo presentó actividad hemolítica. También se observó que presentaron sensibilidad a 6 antibióticos y resistencia a la vancomicina. Por último, los sobrenadantes sin tratamiento de todos los cultivos presentaron actividad antimicrobiana moderada frente a E. coli con halos de inhibición de16 – 19 mm; mientras que, para S. typhi se observó actividad antimicrobiana baja con halos de inhibición de 10 – 14 mm. De los ocho cultivos analizados solo 5 fueron los más prometedores que cumplieron con las características potencialmente probióticas; el análisis filogenético y la secuenciación del ARNr 16S identificaron a todos los aislados como Lactobacillus paracasei subsp. tolerans JCM 1171.
The purpose of this work was to isolate Lactobacillus spp. of artisanal chicha de jora from three producing establishments in the “La Recoleta” retail center in the department of Cajamarca. 31 BAL cultures were isolated, of which only eight met the morphological, staining and biochemical characteristics corresponding to Lactobacillus spp. They were tested for acidic pH resistance, bile salt tolerance, hemolytic activity, gelatin liquefaction capacity, antibiotic susceptibility, and antimicrobial activity to evaluate their probiotic potential. The results obtained showed that at pH 4 all cultures presented survival equal to or greater than 45%, only 6 presented resistance when they were subjected to pH 3 and no culture presented resistance to pH 2. When they were subjected to concentrations of 0.3% and 0.5% of bile salts, all cultures had a survival greater than 90%. In the virulence tests, no culture showed proteolytic activity and only one culture showed hemolytic activity. It was also observed that they presented sensitivity to 6 antibiotics and resistance to vancomycin. Finally, the untreated supernatants of all cultures presented moderate antimicrobial activity against E. coli with inhibition zones of 16 – 19 mm; while, for S. typhi, low antimicrobial activity was observed with inhibition zones of 10 – 14 mm. Of the eight cultures analyzed, only 5 were the most promising that met the potentially probiotic characteristics; phylogenetic analysis and 16S rRNA sequencing identified all isolates as Lactobacillus paracasei subsp. tolerans JCM 1171.
The purpose of this work was to isolate Lactobacillus spp. of artisanal chicha de jora from three producing establishments in the “La Recoleta” retail center in the department of Cajamarca. 31 BAL cultures were isolated, of which only eight met the morphological, staining and biochemical characteristics corresponding to Lactobacillus spp. They were tested for acidic pH resistance, bile salt tolerance, hemolytic activity, gelatin liquefaction capacity, antibiotic susceptibility, and antimicrobial activity to evaluate their probiotic potential. The results obtained showed that at pH 4 all cultures presented survival equal to or greater than 45%, only 6 presented resistance when they were subjected to pH 3 and no culture presented resistance to pH 2. When they were subjected to concentrations of 0.3% and 0.5% of bile salts, all cultures had a survival greater than 90%. In the virulence tests, no culture showed proteolytic activity and only one culture showed hemolytic activity. It was also observed that they presented sensitivity to 6 antibiotics and resistance to vancomycin. Finally, the untreated supernatants of all cultures presented moderate antimicrobial activity against E. coli with inhibition zones of 16 – 19 mm; while, for S. typhi, low antimicrobial activity was observed with inhibition zones of 10 – 14 mm. Of the eight cultures analyzed, only 5 were the most promising that met the potentially probiotic characteristics; phylogenetic analysis and 16S rRNA sequencing identified all isolates as Lactobacillus paracasei subsp. tolerans JCM 1171.
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Keywords
Lactobacillus, probióticos, chicha de jora, tolerancia