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This study aimed to classify a collection of Enterobacter sakazakii ( E. sakazakii) strains previously identified from powdered infant formula (PIF) to species level by recN gene sequencing and biochemical testing to determine the distribution of Cronobacter species in China and investigate the strain diversity by cellular fatty acid (CFA) analysis. Of 24 E. sakazakii isolates, 23 were identified as C. sakazakii and one was C. malonaticus. The 23 C. sakazakii isolates showed the same CFA profiles. The C. malonaticus isolate was discriminated from the C. sakazakii isolates by the significant difference in the amounts of C, C, and C acids. These results showed that C. sakazakii and C. malonaticus were the common Cronobacter species distributed in PIF in China and that the isolates of the two species exhibited different CFA profiles. These findings are of value for epidemiological investigations and provide an alternative method for confirming various Cronobacter spp. [ABSTRACT FROM AUTHOR]