Bifidobacteria have long since been recommended as indicators of human and animal pollution. Concentration ratio (tracking ratio) of the sorbitol-utilising bifidobacteria (SUB) and the total bifidobacteria (TB) can be used to distinguish between animal and human sources of faecal water contamination. The cut-off value needs to be calibrated in a given geographical area. Seven sites with permanent faecal contamination were selected in South Africa. Concentrations of SUB ranged from 10-50000 cells/100 mL, while TB ranged from 0-8000 cells/100 mL. The tracking ratio ranged from 0.10 to 6.25, but no clear cut-off value could be established. The YN-17 agar was replaced for TB with the modified Beerens medium with pH = 5.70, to suppress the growth of faecal streptococci. Tracking ratios observed are most likely the results of different survival rates of SUB and TB. Bifidobacteria die-off due to nutrients was not found to be significant using design of experiment. Thus a lack of continuous input or oxygen levels in water may be major factors. This would limit the ratios used as a faecal source tracking method.