A simple modified membrane filtration medium for the enumeration of aerobic spore-bearing bacilli in water

Aerobic spore-bearing bacilli have been proposed as a surrogate indicator for the removal of Cryptosporidium by drinking water treatment processes. Pasteurisation of samples followed by culture on non-selective media is the method of choice. Using white membranes for filtration of water samples makes colony counting difficult. Vital dyes such as neutral red or trypan blue can help when added to the growth medium but these dyes tend to colour the membrane and reduce the contrast between the colonies and the background. The incorporation of bromothymol blue at a concentration of 0.005% (w/v) into nutrient agar facilitated colony counting without inhibiting colony formation compared with unsupplemented nutrient agar. Statistical analysis of the data (ANOVA) confirms this observation. The modified technique was found to be satisfactory with spore suspensions of Bacillus globigii and B. cereus as well as with samples of surface water, settled water and drinking water.