Listeria Chromogenic Medium is used
for rapid detection of Listeria monocytogenes in food and medicine. Negative
results can be detected completely in three days. The colonies of Listeria
monocytogenes is blue, surrounded by an opaque ring. On recipe, peptone
provides nitrogen and amino acids, sodium chloride maintains osmotic pressure,
agar is coagulants, lithium chloride inhibits the growth of Gram-negative
bacteria, bacteriosestatic agents inhibits the growth of Gram-positive bacteria,
except Listeria.
Ingredients(g/L):
Nutrients
|
70.0
|
Chromogenic reagent
|
1.0
|
Final pH 7.2 ± 0.2 at 25℃
|
Additives:
Listeria
additive 6.0g
Listeria
bacteriosestatic agent 1 1mL* 5 pieces
Listeria
bacteriosestatic agent 2 1mL * 5 pieces
*Adjusted and/or
supplemented as required to meet performance criteria.
Directions:
Suspend 14.2g of the medium in 180ml distilled water, and weigh 1.2g
Listeria additive, add 20ml of sterile water and some glass balls, , both are
autoclaved at 121 ℃ for 15 min. When cooling down to about 50 ℃, put the Listeria additive into the base medium, and
add each one piece of Listeria antibacterial agents 1 and 2, mix evenly, pour
into sterile petri dishes. Medium thickness is about 5mm, so advantageous to
observe opaque ring.
Procedures:
Before to use, the prepared medium is placed
at room temperature for 10-20 minutes
1. Prepared sample solution according to
national standards, SN standard, FDA standards or other methods.
2. Put samples solution into Listeria
enrichment broth (LB1, LB2) for enrichment culture; or the sample solution and
the Half-Fraser enrichment broth according to the mixing ratio of 1:10, 30 ℃ for 24 hours; Or other enrichment broth for
enrichment culture;
3. Take one ring enrichment broth, streak
and inoculate on Listeria chromogenic medium, culture for 24-48 hours at 37 ℃.
Quality Control:
1. Appearance:
The dehydrate powder medium is homogeneous,
having good flowability; yellowish, prepared medium is opaque milky white
solid.
2. Microbial sensitivity test:
Incubated at 37 ℃ for 24-48 hours:
QC Strains
|
Strains No.
|
Growth
|
Colonies color
|
Opaque ring
|
Listeria
monocytogenes
|
ATCC27853
|
+++
|
blue
|
+
|
Listeria iuanuii
|
CICC21663
|
+++
|
blue
|
+
|
Listeria seeligeri
|
CICC21617
|
+++
|
blue
|
+
|
Staphylococcus
aureus
|
ATCC25923
|
++
|
yellow
|
+
|
E. coli
|
ATCC25922
|
-
|
-
|
-
|
salmonella
|
ATCC14028
|
-
|
-
|
-
|
Typical features:
Listeria monocytogenes culture for 24-28
hours at 37 ℃, each of blue colonies are surround by an
opaque ring. Listeria iuanuii culture for 48 hours at 37 ℃, each of blue colonies are surrounded by an opaque
ring. Listeria seeligeri culture for 24-48 hours at 37 ℃, blue colonies appear, no opaque ring around. Listeria
iuanuii inspection is very rare in food.
Storage:
The prepared plates can be kept for 2-8 ℃ avoid direct exposure to light. The powder medium
should be placed in a dark, dry place, storage temperature 2-8 ℃, avoid direct exposure to light.
Failure:
Dry powder media which are over the shelf
life, caking and color variation cannot be used.
Note:
This medium is only for laboratory use.
Reference:
1. VAN NETTEN, P., PERALES, J., VAN DE
MOOSDIJK, A., CURTIS, G.D.W., a. MOSSEL, D.A.A.: Liquid and solid selective
differential media for the detection and enumeration of Listeria
monocytogenes. Int. Food Microbiol., 8;
299-316 (1989).
2. HAMMER, G., HAHN, G., KIRCHHOFF, H., a.
HEESCHEN, W.: Vergleich der Eignung von Oxford- und PALCAM-Medium zur
Isolierung von Listeria monocytogenes aus Weichkäse. Dtsch. Milchwirtschaft,41; 334-336 (1990).