SCHEMATIC DIAGRAM

ISOLATION AND ENUMERATION OF LISTERIA

1. FOOD
HOMOGENATE




2. PRE-ENRICHMENT




3. ISOLATION








4. READING


5. CONFIRMATION


25 gram sample

225 mL Listeria enrichment broth
Blend at 10,000-12,000 rpm for 2 min
Listeria colonies are black with
halo on esculin-containing media


After 4 h at 30
o
C in buffered Listeria
Enrichment Broth (LEB), add selective
agents acriflavin, 10mg/L, sodium
naliixate, 40mg/L, cycloheximide
50mg/L or natamycin, 25mg/L
FOOD ANALYTICAL SERVICE LABORATORY
FOOD AND NUTRITION RESEARCH INSTITUTE
MIC-AM-007-
Incubate at 30
o
C
for 24-48 hr; streak
plates at 24 hr and
after 48 hr

Streak BLEB
onto OXA.
Incubate at 35
o
C
for 24-48 h

Use rapid kits for biochemical testing to genes or species level.

Transfer 5 or more typical colonies from OXA to trypticase soy agar with yeast
extract (TSAYE) streaking for purity, incubate at 30
o
C, 24-48 h. Plates may be
incubated at 35
o
C if colonies are not used for wet mount motility.
oxa oxa
or
PALCAM
agar
PALCAM
agar

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Effectivity Date:
May 15, 2012