Slanetz and Bartley HiCynth™ Medium

Principle And Interpretation

Slanetz and Bartley Medium was originally devised by Slanetz and Bartley (1) for the detection and enumeration of Enterococci by membrane filtration technique. It can be also used as a direct plating medium (2, 3). The medium is highly selective for Enterococci. The medium is modified by using chemically defined peptones instead of animal peptones to avoid BSE/TSE risk.

HiCynth™ Peptone No.1 and HiCynth™ Peptone No.5 in the medium provide the necessary nitrogen and carbon compounds, long chain amino acids, vitamins and minerals required for the growth of organisms. Sodium azide has inhibitory effect on gram-negative organisms. Triphenyl Tetrazolium Chloride is reduced to the insoluble formazan inside the bacterial cell forming dark red-coloured colonies. When the medium is incubated at higher temperature (44-45°C), all red or maroon colonies can be considered as presumptive Enterococci (4, 5).

The Department of Health (6) has recommended this medium to be used for enumeration of Enterococci in water supplies. Water is filtered through a membrane filter which is then placed on the surface of the Slanetz and Bartley HiCynth™ Medium plates and incubated at 35°C for 4 hours and then at 44-45°C for 44-48 hours. Red or maroon colonies are counted as Enterococci. The preliminary incubation at 35°C helps for the recovery of stressed organisms. Not all the species reduce TTC, hence pale colonies also should be considered.

Food samples are homogenized and so diluted with physiological saline to give 15-150 colonies on each Petri plate. Homogenates or dilutions are spread on agar surface and incubated at 35°C for 48 hours. Pink or dark red colonies with a narrow whitish border are counted (3).