A simple medium modification for isolation, growth and enumeration of Acidithiobacillus thiooxidans (syn. Thiobacillus thiooxidans) from water samples.

High concentrations of H(2)S in groundwater are commonly removed using Biological Trickling Filter (BTF) that contains high numbers of biofilm immobilized sulfur oxidizing bacteria (mainly Thiobacillus thiooxidans). BTF performance requires continuous monitoring of these bacteria at several sampling points. The Most Probable Number (MPN) technique is at the moment the method of choice to enumerate viable T. thiooxidan cells under the above conditions. However, this method is extremely time-consuming (7-10days) and not always suitable for environmental monitoring. In the present study, Thiobacillus agar recommended for isolation and cultivation of Thiobacillus species by Spread plate method was modified by addition of bromocresol green (BCG) in order obtain a clear-cut resolution of the growing colonies resulting in similar or higher numbers compared to other methods. Visual emergence of bacterial colonies on the 3rd and 4th days, from the initial plating, was associated with sulfuric acid production, resulting in an unambiguous color change from blue to yellow, around each colony. This study revealed that BCG modified Thiobacillus agar is substantially time saving and much easier to infer compared to MPN technique.