A Method of Screening for Highly Inhibitory Lactic Acid Bacteria

Abstract

A rapid method of screening for lactic acid bacteria with high inhibitory power was developed. The methodology employed was the agar-diffusion assay, which was standardized for the indicator strain, medium composition, and incubation conditions. The assay was performed in nutrient agar at 30°C with 108 spores of Bacillus subtilis per plate as indicator strain. The inhibition produced by supernatants of lactic acid bacteria cultures harvested at stationary phase was determined. The inhibitory powers of different strains were compared with a standard curve obtained with racemic lactic acid. Results obtained with lactic acid and supernatants of bacterial cultures demonstrate that the diameter of the inhibition zone (d) was related to the pH by the exponential relation d = a exp (− b pH). Results obtained with strains that produced inhibitory substances other than lactic acid could not be fitted into the standard linear curves obtained in the plot of ln d versus pH of lactic acid.