Absence of oxygen affects the capacity to sporulate and the spore properties of Bacillus cereus

• Bacillus cereus sporulation was low and heterogeneous in anaerobiosis.
• Spores from anaerobiosis were more resistant to wet-heat, NaOH and nitrous acid.
• Germination was more efficiently in anaerobiosis compare to aerobiosis.
• MET of AH 187 spores from anaerobiosis showed a damaged exosporium.

This study was performed to evaluate the effect of anaerobiosis on the formation of Bacillus cereus spores and their resulting properties. For this purpose, an appropriate sporulation medium was developed (MODs). Sporulation of 18 strains from different phylogenetic groups of B. cereus was studied in MODs medium in aerobiosis and anaerobiosis. In anaerobiosis, sporulation ability was weaker and more heterogeneous than in aerobiosis. Among tested strains, B. cereus AH187 produced the highest level of spores in anaerobiosis. This strain was therefore chosen to study spore properties. Spores produced in anaerobiosis were more resistant to wet heat at 90 °C, 92.5 °C, 95 °C, 97.5 °C and 100 °C. For example, D90 were 21,09 ± 1.70 and 81.87 ± 2.00 for aerobiosis and anaerobiosis conditions, respectively. Spores produced in anaerobiosis have a z-value of 7.70 °C compared with 10.52 °C for spores produced in aerobiosis. Spores produced in anaerobiosis were also more resistant to 1 M NaOH, 1 M nitrous acid and pulsed light at fluences of 0.34 J cm−2 and 0.49 J cm−2. No difference in resistance to UV-C, 5% hydrogen peroxide or 0.25 mM formaldehyde was observed between these two conditions. In the presence of l-alanine, spores produced in anaerobiosis germinated more efficiently than spore produced in aerobiosis. No difference in germination was observed with inosine as inducer. No difference in the size of spores produced in the different conditions was observed by transmission electron microscopy. However, spores obtained under anaerobic conditions had a damaged exosporium, or in some cases a completely detached exosporium, unlike spores produced under aerobic conditions.This study shows that few spores are formed under anaerobic condition; nevertheless, this condition has an impact on the spore properties of B. cereus AH 187 strain. Spores obtained under anaerobic condition were more resistant to heat and to some chemical compounds. This is an important feature, considering the risk associated with the presence of this pathogen in thermally processed and packaged food in absence of oxygen.