Abstract
Abstract method was developed for detection of Bacillus cereus spores in milk. It includes heat treatment at 72°C, followed by treatment with trypsin and Triton-X100 at 55°C, and membrane filtration. Colonies of B. cereus could be recognised by their typical colony morphology and the zone of hemolysis surrounding the colonies following incubation on blood agar with 10 ppm polymyxin. Blood agar was superior to MYP and PEMBA for detection. Psychrotrophic as well as mesophilic strains of B. cereus gave clearly visible micro-colonies after 48 h of incubation at 20°C. At 20°C interference by growth of other hemolytic Bacillus sp. was strongly reduced compared to incubation at 30°C. Heat treatment of the milk at 72°C for 5 min as compared to 75°C for 15 min improved the filterability of the milk and resulted in significantly higher recovery of psychrotrophic spores. At least 100 mL and up to 250 mL of milk could be filtered. Recovery of spores was typically 85±13% (SEM) with a mean coefficient of variation for duplicate analyses of 18%.
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