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Applied and Environmental Microbiology, January 2007, p. 48-52, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.01562-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Comparison of Media for the Isolation of Enterobacter sakazakii{triangledown}

Carol Iversen and Stephen J. Forsythe*

School of Biomedical and Natural Sciences, Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS, United Kingdom

Received 6 July 2006/ Accepted 13 October 2006

Enterobacter sakazakii is associated with neonatal infections and is occasionally present at low levels (<1 CFU/g) in powdered infant formula milk (IFM). It has been previously reported that some E. sakazakii strains do not grow in standard media for Enterobacteriaceae and coliform bacteria; therefore, a reliable method is needed for recovery of the organism. Three E. sakazakii enrichment broths—Enterobacteriaceae enrichment broth (EE), E. sakazakii selective broth (ESSB), and modified lauryl sulfate broth (mLST)—were compared with a novel broth designed for maximum recovery of E. sakazakii, E. sakazakii enrichment broth (ESE). One hundred seventy-seven strains (100%) grew in ESE, whereas between 2 and 6% of strains did not grow in EE, mLST, or ESSB. E. sakazakii possesses {alpha}-glucosidase activity, and a number of selective, chromogenic agars for E. sakazakii isolation based on this enzyme have been developed. E. sakazakii isolation agar produced fewer false-positive colonies than did Druggan-Forsythe-Iversen agar. However, the latter supported the growth of more E. sakazakii strains. It was also determined that 2% of E. sakazakii strains did not produce yellow pigmentation on tryptone soya agar at 25°C, a characteristic frequently cited in the identification of E. sakazakii. The recovery of desiccated E. sakazakii (0.2 to 2000 CFU/25 g) from powdered IFM in the presence of a competing flora was determined with various enrichment broths and differential selective media. Current media designed for the isolation and presumptive identification of E. sakazakii do not support the growth of all currently known E. sakazakii phenotypes; therefore, improvements in the proposed methods are desirable.

* Corresponding author. Mailing address: School of Biomedical and Natural Sciences, Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS, United Kingdom. Phone: 1158483529. Fax: 1158486636. E-mail: stephen.forsythe{at}ntu.ac.uk.

{triangledown} Published ahead of print on 27 October 2006.

Applied and Environmental Microbiology, January 2007, p. 48-52, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.01562-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Almeida, C., Azevedo, N. F., Iversen, C., Fanning, S., Keevil, C. W., Vieira, M. J. (2009). Development and Application of a Novel Peptide Nucleic Acid Probe for the Specific Detection of Cronobacter Genomospecies (Enterobacter sakazakii) in Powdered Infant Formula. Appl. Environ. Microbiol. 75: 2925-2930 [Abstract] [Full Text]  
  • Iversen, C., Druggan, P., Schumacher, S., Lehner, A., Feer, C., Gschwend, K., Joosten, H., Stephan, R. (2008). Development of a Novel Screening Method for the Isolation of "Cronobacter" spp. (Enterobacter sakazakii). Appl. Environ. Microbiol. 74: 2550-2553 [Abstract] [Full Text]  
  • Townsend, S. M., Hurrell, E., Gonzalez-Gomez, I., Lowe, J., Frye, J. G., Forsythe, S., Badger, J. L. (2007). Enterobacter sakazakii invades brain capillary endothelial cells, persists in human macrophages influencing cytokine secretion and induces severe brain pathology in the neonatal rat. Microbiology 153: 3538-3547 [Abstract] [Full Text]  


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