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The prevalence and characteristics of low-temperature-tolerant Bacillus cereus (psychrotolerant B. cereus ) in green leaf lettuce collected during cold chain were investigated. Among the 101 isolated B. cereus samples, only 18 were capable of growth at 7 °C, and these isolates shared potential health hazard characteristics with mesophilic isolates. Most psychrotolerant B. cereus isolates contained various combinations of nheA, nheB, nheC,  hblA, hblA, hblC, hblD , cytK, and entFM. Most isolates of psychrotolerant B. cereus possessed at least two enterotoxin genes and 28% of isolates harbored tested nine enterotoxin genes. Additionally, the psychrotolerant B. cereus isolates showed resistance to tetracycline and rifampin and intermediate levels of resistance to clindamycin. A total of 23% of isolates among psychrotolerant B. cereus displayed a high level of biofilm formation at 7 °C than at 10 °C or 30 °C. The results of this study indicate that cold distribution and storage for green leaf lettuce may fail to maintain food safety due to the presence of enterotoxigenic, antibiotic-resistant, and strong biofilm forming psychrotolerant B. cereus isolates, which therefore poses a potential health risk to the consumer. Our findings provide the first account of the prevalence and characteristics of psychrotolerant B. cereus isolated from green leaf lettuce during cold storage, suggesting a potential hazard of psychrotolerant B. cereus isolates to public health and the food industry.

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Enterotoxin Genes, Antibiotic Susceptibility, and Biofilm Formation of Low-Temperature-Tolerant Bacillus cereus Isolated from Green Leaf Lettuce in the Cold Chain