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Chapter 2 : Isolation and Identification of

Authors: Séamus Fanning1, Steve Forsythe2
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Affiliations: 1: Centre for Food Safety, School of Agriculture, Food Science, and Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland; 2: School of Biomedical and Natural Sciences, Nottingham Trent University, Nottingham, United Kingdom
Content Type: Monograph
Publication Year: 2008

Category: Applied and Industrial Microbiology

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Isolation and Identification of , Page 1 of 2

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Abstract:

Due to the association of with neonatal infections, the organism has primarily been associated with powdered infant formula. However, is widely distributed in the environment and in foods, with the most probable sources of the organism being water, soil, and vegetables. It is important that infection from rehydrated infant formula can arise from contaminated water, introduction by the caregiver, or the environment. In addition to specific criteria for , the replacement of the original indicator organism group, coliforms with , has been proposed. The Food and Drug Administration (FDA)-approved method is a presumptive, most-probable-number test based on the three-tube enrichment format, so that low levels of the organism, if present in the product, can be detected and quantified. A chromogenic agar was developed, designated Druggan-Forsythe-Iversen (DFI) agar, where a chromogenic substrate, 5-bromo-4-chloro-3- indolyl-α-D-glucopyranoside, is used as an indicator of α-glucosidase activity. Rapid development of laboratory detection and recognition methods for infectious agents has been aided by the rapid and innovative developments in nucleic acid amplification technologies. Molecular subtyping of bacteria by profiling either proteins or nucleic acids is a useful approach to investigate the epidemiological relationship(s) of isolates involved in outbreaks of food-borne disease. A multilocus sequence typing method using 10 housekeeping genes is presently under development for and looks very promising as a molecular typing tool for the epidemiology of .

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
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Isolation and Identification of Enterobacter sakazakii
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Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02fig01
Figure 1
Conventional cultural detection of (from Forsythe, ).
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10.1128/9781555815608/f0035-01.gif
Image of Figure 1
Figure 1

Conventional cultural detection of (from Forsythe, ).

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
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Figure 2
XbaI-generated PFGE profiles for a collection of isolates.
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10.1128/9781555815608/f0049-01.gif
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Figure 2

XbaI-generated PFGE profiles for a collection of isolates.

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
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Figure 3
ERIC-PCR DNA profiles for a collection of isolates and a strain.
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Image of Figure 3
Figure 3

ERIC-PCR DNA profiles for a collection of isolates and a strain.

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
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Tables

Isolation and Identification of Enterobacter sakazakii
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Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab01
Table 1
Sources of
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Table 1

Sources of

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab02
Table 2
Summary of cultured from different infant formulations
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Table 2

Summary of cultured from different infant formulations

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab03
Table 3
Surveys of powdered infant formula or milk powders for
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Table 3

Surveys of powdered infant formula or milk powders for

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab04
Table 4
General microbial flora cultured from powdered infant formula
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Table 4

General microbial flora cultured from powdered infant formula

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab05
Table 5
Proposed CCFH and ICMSF microbiological criteria
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Table 5

Proposed CCFH and ICMSF microbiological criteria

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab06
Table 6
Presence of in samples in which were present in or absent from powdered infant formula samples
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Table 6

Presence of in samples in which were present in or absent from powdered infant formula samples

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab07
Table 7
Culture methods for the detection of in powdered infant formula
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Table 7

Culture methods for the detection of in powdered infant formula

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab08
Table 8
Composition of enrichment broths
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Table 8

Composition of enrichment broths

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab09
Table 9
Composition of selective and differential agars for
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Table 9

Composition of selective and differential agars for

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab10
Table 10
Percentage of strains showing increase in optical density after 24 h of incubation in enrichment media
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Table 10

Percentage of strains showing increase in optical density after 24 h of incubation in enrichment media

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab11
Table 11
Percent growth of strains and other on various selective and differential agars
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Table 11

Percent growth of strains and other on various selective and differential agars

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab12
Table 12
Comparison of sensitivity and specificity of the FDA method with ISO (DTS 22964) for the isolation of
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Table 12

Comparison of sensitivity and specificity of the FDA method with ISO (DTS 22964) for the isolation of

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab13
Table 13
Sensitivity and specificity of three biochemical profiling methods
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Table 13

Sensitivity and specificity of three biochemical profiling methods

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab14
Table 14
biotypes and 16S rRNA gene sequence cluster groups
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Table 14

biotypes and 16S rRNA gene sequence cluster groups

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2
/content/10.1128/9781555815608.ch02.ch02tab15
Table 15
PCR primers and their thermodynamic characteristics, genome targets, and sequences used for the detection of
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Table 15

PCR primers and their thermodynamic characteristics, genome targets, and sequences used for the detection of

Citation: Fanning S, Forsythe S. 2008. Isolation and Identification of , p 27-59. In Farber J, Forsythe S, Doyle M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815608.ch2

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