Typing and evaluating heat resistance of Bacillus cereus sensu stricto isolated from the processing environment of powdered infant formula

Abstract

Bacillus cereus sensu lato is one of the most harmful bacterial groups affecting the quality and safety of powdered infant formula (PIF). In this study, samples were collected from the raw materials and processing environments of PIF. A total of 84 isolates were identified as Bacillus cereus sensu stricto (B. cereus s. s.) by 16S rRNA analysis, molecular typing technology, and physiological and biochemical tests. The 84 B. cereus s. s. strains were assigned to panC group II, group III, and group IV. Then, the 7 housekeeping genes glpF, gmk, ilvD, pta, pur, pycA, and tpi were selected for multilocus sequence typing. Results showed that the 84 isolates were clustered into 24 sequence types (ST), and 14 novel ST were detected. Among the 24 ST, ST999 (19/84, 22.62%) and ST1343 (13/84, 15.48%) predominated. The correlation between processing areas and ST showed that the processing environments of the production and packing areas were the most susceptible to contamination by B. cereus s. s. Spores of these ST showed different heat resistance phenotypes evaluated by the analysis of DT (time in minutes of spore decimal reduction at each temperature) and Z values (temperature increase required to reduce the DT value to one-tenth of the original). Spores from group III according to panC gene analysis were the most heat resistant. These findings will help us to better understand B. cereus s. s. contamination and control in PIF processing environments.