Two outbreak-related Bacillus cereus emetic strains were investigated for their growth and cereulide production potential in penne pasta at 4, 8 and 25 °C during 7-day storage. Cereulide production was detected and quantified by LC-MS method (LOD of 1 ng/ml, LOQ of 5 ng/ml) and growth was determined by culture-based enumeration. Inoculated B. cereus strains (10(5) CFU/g) were able to reach counts of more than 10(8) CFU/g and cereulide production of about 500 ng/g already after 3 days of storage at 25 °C. Interestingly, a constant increase of the toxin was noticed during incubation at ambient temperature storage: the cereulide was continuously produced during the bacterial stationary growth phase reaching maximal amounts at the end of the experiment (7 days, concentration of about 1000 ng/g). Strictly respected cold chain temperature as 4 °C did not allow any detectable cereulide production for any of the two tested strains. At the limited temperature abuse of 8 °C, a detectable amount of cereulide was observed after two days for one of the strain (TIAC303) (<LOQ). These results confirm that cereulide production is controlled by multiple factors (from type of strain to temperature) and that prolonged storage time plays a crucial role for consumer safety.
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