Evaluation of Selective Media Containing Iron Source and Alpha-Glucosidase Substrates for Enterobacter sakazakii (Cronobacter spp.) Detection

Enterobacter sakazakii (Cronobacter spp.) causes meningitis, necrotizing enterocolitis, sepsis, and bacteremia in neonates and children and has a high mortality rate. For rapid E. sakazakii detection, various differential and selective media containing α-glucosidase substrates, such as 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside (BCIG) or 4-methylumbelliferyl-α-D-glucoside (α-MUG), have been developed as only E. sakazakii exhibits α-glucosidase activity in the genus Enterobacter. However, Escherichia vulneris (family: Enterobacteriaceae) can also utilize α-glucosidase substrates, thereby resulting in false positives. Various iron sources are known to promote the growth of gram-negative bacteria. This study aimed to develop a selective medium containing α-glucosidase substrates for E. sakazakii detection that would eliminate false positives, such as those of E. vulneris, and to determine the role of iron source in the medium. Three previously developed (TPD) media, i.e., Oxoid, OK, and VRBG, and the medium developed in this study, i.e., NGTE, were evaluated using 58 E. sakazakii and 5 non-E. sakazakii strains. Fifty-four E. sakazakii strains appeared as fluorescent or chromogenic colonies on all four media that were assessed. Two strains showed colonies on NGTE medium and not on TPD media. In contrast, the remaining two strains showed colonies on TPD media and not on NGTE medium. None of the non-E. sakazakii strains showed fluorescent or chromogenic colonies on any of the evaluated media except E. vulneris, which showed colonies on TPD media and not on NGTE medium. This study demonstrated that the newly developed NGTE medium was not only equally efficient in promoting the growth of bacterial colonies when compared with the currently available media but also eliminated false positives, such as E. vulneris.