Buffered HiCynth™ Peptone Water

Principle And Interpretation

Buffered HiCynth™ Peptone Water is a pre-enrichment medium designed to help recovery of sub-lethally damaged Salmonellae before transfer to a selective medium. It is prepared by completely replacing animal or vegetable peptones with chemically defined peptones to avoid BSE/TSE risks associated with animal peptones. This pre-enrichment medium is free from inhibitors and is well buffered and provides conditions for resuscitation of the cells that have been injured by processes of food preservation. It was noted by Edel and Kampelmacher (1) that sub-lethal injury to Salmonella may occur due to food preservation techniques involving heat, desiccation, high osmotic pressure, preservatives or pH changes. Buffered HiCynth™ Peptone Water during the pre-enrichment period helps in recovery of injured cells that may be sensitive to low pH (2). This is particularly important for vegetable specimens, which have low buffering capacity. This medium can be used for testing dry poultry feed (3). In a survey involving isolation of Salmonellae from meat that had been artificially contaminated with sub-lethally injured organisms, pre-enrichment in Buffered Peptone Water at 37°C for 18 hours before selection in Tetrathionate Brilliant Green Bile Broth showed superior results compared with direct selection method. Lactose Broth is frequently used as a pre-enrichment medium but it may be detrimental to recovery of Salmonellae (4).

The media contain HiCynth™ Peptone No.1 as a source of carbon, nitrogen, vitamins and minerals. Sodium chloride maintains the osmotic balance and phosphates buffer the medium. The broth is rich in nutrients and produces high resuscitation rates for sublethally injured bacteria and supports intense growth. The phosphate buffer system prevents bacterial damage due to changes in the pH of the medium.

Inoculate 10 grams specimen in 50 ml of these media and incubate at 35-37°C for 18 hours. Transfer 10 ml from this medium to 100 ml of Tetrathionate Broth and incubate at 43°C for 24 - 48 hours and then subculture on selective plating media. Examine the plates for characteristic Salmonella colonies.