• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.579-584
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• 2008

A commercial chromogenic agar medium (DFI) was supplemented with glucose (mDFI) to enhance the specificity of Enterobacter sakazakii (E. sakazakit) detection. Escherichia vulneris (E. vulneris), a putative false-positive strain on the DFI medium, produces ${\alpha}$-glucosidase. The enzyme ${\alpha}$-glucosidase hydrolyzes a substrate, 5-bromo-4-chloro-3-indolyl-${\alpha}$, D-glucopyranoside $(X{\alpha}Glc)$, producing green colonies. E. sakazakii strains produced green colonies on both DFI and mDFI agar, whereas E. vulneris produced green colonies on DFI agar but small white colonies on mDFI agar. E. sakazakii and E. vulneris were also readily differentiated by colony color when the mixed culture of the two strains was plated on mDFI agar and incubated for 24 h at $37^{\circ}C$. The results indicate that the selectivity of the commercial chromogenic agar medium could be improved by a simple supplementation with glucose.

• Journal of Dairy Science and Biotechnology
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• v.39 no.1
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• pp.9-19
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• 2021

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.

• Food Science of Animal Resources
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• v.28 no.5
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• pp.555-561
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• 2008

Ninety-nine samples of powdered infant formula in a market were collected from the local market and their contaminations for total aerobic bacteria, coliform, FAO/WHO Category A, B, and C pathogens were analyzed. Total aerobic bacteria were detected in 92 of 99 samples (93%) at levels of $1.83{\pm}0.68\;Log\;MPN/g$. These levels were below legal levels specified for infant formulas except for one sample detected by 4.5 Log CFU/g. Coliform was detected in 12 of 99 samples (12%) at levels of $1.26{\pm}1.03\;Log\;MPN/g$ whereas non-detection was required according to the specification of coliform in infant formulas. Escherichia coli was detected in 1 of 99 samples by 0.48 Log MPN/g. Salmonella and Enterobacter sakazakii among Category A weren't detected in all the samples. Enterobacteriaceae, Category B group, were detected in 25 samples of total 99 samples (25%) by $0.83{\pm}1.37\;Log\;MPN/g$. Enterobacteriaceae identified by API 20E were Escherichia vulneris, Es. hermannii, Pantoea spp., Citrobacter koseri, Klebsiella pneumoniae, En. cloaceae. Bacillus cereus among Category C was highly detected in 29 of 99 samples (29%) at levels of $0.69{\pm}0.32\;Log\;MPN/g$ with the most probable number count method, which were below legal levels for the specification of B. cereus in infant formulas. Clostridium perfringens, E. coli O157, Staphyloccus aureus, Listeria monocytogenes, Yersinia enterocolitica, and Campylobacter jejuni/coli were not detected. Contamination level of major pathogens was low and falls within the range of specification of infant formulas. However, Enterobacteriaceae and B.cereus showed the high prevalence and some Enterobacteriaceae causing disease were detected. Therefore, it is necessary to monitor the potential pathogens continually and reduce them to improve the microbial quality of non-sterilized powdered infant formulas.