Evaluation of a New One-Step Enrichment in Conjunction with a Chromogenic Medium for the Detection of Cronobacter spp. (Enterobacter sakazakii)in Powdered Infant Formula

Abstract

The aim of the present study was to evaluate a new one-step enrichment protocol, consisting of a combined preenrichment and enrichment broth (Cronobacter Enrichment Broth [CEB]) used in conjunction with selective-differential agar ChromID Sakazakii, to facilitate a shortened 2-day cultural method for detection of Cronobacter spp. (Enterobacter sakazakii ) in powdered infant formula (PIF). The CEB was evaluated using samples artificially inoculated with low concentrations of 10 lyophilized strains, representative of the genus Cronobacter. The detection of strains was compared in parallel with the enrichment medium from ISO/TS 22964 and a recently proposed differential screening broth for the detection of Cronobacter. All of the Cronobacter strains were recovered using the CEB, and a significantly higher final bacterial concentration was obtained with the CEB than with the other enrichment broths (P < 0.01). There was no significant difference between the cell concentrations for cultures grown in CEB at 37°C and those grown at 41.5°C. Cronobacter was recovered from both 1/10 (50 g:450 ml) and 1/5.5 (100 g:450 ml) sample-to-broth ratios, with no significant difference observed between the final bacterial concentrations obtained from the two ratios. Further studies on a wider range of PIFs, including naturally contaminated samples, are warranted to determine if the use of this protocol may facilitate the rapid release (within 40 to 48 h) of PIF.