472 Handbook of Culture Mediafor Food Microbiology, J.E.L. Corry et al. (Eds.

)
9 2003 Elsevier Science B.V. All rights reserved

Fraser broth and modified Half Fraser broth

This monograph has been reviewed by members of the IUMS-ICFMH Working Party
on Culture Media and given 'Proposed' status.

Description and history

Fraser and Sperber (1988) developed the secondary enrichment broth (UVM Listeria
Enrichment Broth II) used in the United States Department of Agriculture procedure
for isolating Listeria monocytogenes from meats (McClain and Lee, 1988). The
present formula is a subsequent modification with an increased amount of acriflavine
(Cook, 1998). It gives a presumptive test for Listeria spp. as it contains an indicator
for the hydrolysis of aesculin. This reaction is not exclusive to Listeria spp. so any
microorganisms giving a positive reaction (blackening) must be isolated and further
identified. The 'Half Fraser' broth used as a primary enrichment in the ISO (1997)
method contains reduced amounts of the selective agents nalidixic acid and acriflavine
to aid the recovery of injured cells.

Composition (grams)

Proteose peptone (peptic digest of animal tissue) 5.0

Tryptone (pancreatic digest of casein) 5.0
Beef extract 5.0
Yeast extract 5.0
Sodium chloride 20.0
Di-sodium hydrogen orthophosphate .2H20 12.0
Potassium di-hydrogen orthophosphate 1.35
Aesculin 1.0
Lithium chloride 3.0
Iron (III) ammonium citrate 0.5
Nalidixic acid 0.02 a or 0.01 b
Acriflavine 0.025 a or 0.0125 b
Distilled or deionized water 1000.0

a Fraser broth
b Half Fraser broth
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Preparation

Basal broth

Suspend all of the ingredients, except the acriflavine and iron (III) ammonium citrate,
in the water and heat to dissolve. Dispense 10 ml portions into 16 x 150 mm culture
tubes with screw cap closures and sterilize at 121~ for 15 min.

Supplements

To each tube of the cooled basal broth, immediately before use, add 0.1 ml portions of
filter sterilized aqueous solutions of 0.25% of acriflavine and 5.0% iron (III) ammo-
nium citrate. For Half Fraser broth add 0.05 ml of 0.25% acriflavine and 0.1 ml of
5.0% iron (III) ammonium citrate.

Physical properties

Appearance Light straw colour, clear.

pH 7.2 + 0.2

Shelf life

Basal broth and 14 days at 4 + 2~ Use complete medium

supplement immediately after addition of the supplements.

Inoculation method for samples

Food or environmental samples are cultured in a primary listeria enrichment broth

(e.g. Half Fraser) and incubated at 30~ for 24 h. These cultures are gently mixed and
0.1 ml portions are added to 10 ml of complete Fraser broth.

Incubation

Half Fraser: at 30~ for 24 h.

Fraser: at 35~ for 24 and 48 h (Warburton et al., 1991), in air and in the dark.

Reading of results and interpretation

After incubation the tubes are compared to an uninoculated control against a white
background. Blackened cultures are considered as presumptively positive for Liste-
ria spp. However, confirmatory tests are necessary on all positive cultures. Cultures
which retain the original straw colour are considered to be negative.
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Quality assessment

(i) Productivity
Test strains Listeria monocytogenes serovar 1/2a (ATCC 35152/
N C T C 7973)
Listeria monocytogenes serovar 4b (ATCC 13932/
NCTC 10527)
Listeria ivanovii serovar 5 NCIMB 50095

Inoculation method Dilution to extinction.

(ii) Selectivity
Test strains Enterococcus faecalis NCIMB 50030
Proteus mirabilis (ATCC 29906/CECT 4168/NCTC
11938)

Inoculation method Dilution to extinction.

Comments

Selective properties of acriflavine may vary from lot to lot and manufacturer to manu-
facturer. Each new batch must be assayed in combination with other selective agents
used in the medium to determine the optimum concentration for use, with regard to
the efficiency of selectivity and absence of inhibition of Listeria spp. Storage of stock
solutions of acriflavine for periods of more than one month is not recommended.

References

Cook, L.V. (1998) Chapter 8. Isolation and identification of Listeria monocytogenes from red meat,
poultry, eggs and environmental samples (Revision 2; 11/08/99) In: USDA/FSIS Microbiology
Laboratory Guidebook 3rdEdn. US Department of Agriculture, Washington, DC.
Fraser, J.A. and Sperber, W.H. (1988) Rapid detection of Listeria spp. in food and environmental
samples by esculin hydrolysis. J. Food Protect. 51,762-765.
ISO (1997) Microbiology of food and animal feeding stuffs - Horizontal method for the detection
and enumeration of Listeria monocytogenes. Part 1. Detection method. ISO 11290-1:1997.
International Organisation for Standardisation, Geneva.
McClain, D. and Lee, W.H. (1988) Development of USDA-FSIS method for isolation of Listeria
monocytogenes from raw meat and poultry. J. Assoc. Off. Anal. Chem. 71,660-664.
Warburton, D.W., Farber, J.M., Armstrong, A., Caldeira, R., Tiwari, N.P., Babiuk, T., Lacasse, P. and
Read, S. (1991) A Canadian comparative study of modified versions of the 'FDA' and 'USDA'
methods for the detection of Listeria monocytogenes. J. Food Protect. 54, 669-676.