Coliforms Chromogenic Medium is used for
fast, accurate detection of coliforms in food, water, milk, ice cream and meat,
culture for 24 hours, the colonies of coliforms are blue-green, other bacteria
are yellow or colorless, gram-positive bacteria are inhibited.
Ingredients (g/L):
Special
Nutrients
|
34.55
|
Chromogenic
reagent
|
0.35
|
Agar
|
13.0
|
Final pH 7.0-7.2
at 25℃
|
*Adjusted
and/or supplemented as required to meet performance criteria.
Directions:
Suspend 47.9g of the medium in 1L of distilled water, boil in 1 minute,
dispensing into flasks, at 121 ℃ autoclaving for 15 minutes, add 1mL
prepared sample into a sterile in 9cm petri dish, transfer 15mL chromogenic
medium when cooling down to 45-50 ℃, mix, inverted culture at 36 ± 1 ℃ for 18-24h after solidification.
Or when cooling down to 45-50 ℃, pour into a sterile petri dish, after
complete solidification, inverted culture at 37 ℃ in incubator for 1-2 hours, add 1mL
uniform liquid sample with a suitable concentration, streak onto the medium ,
culture at 36 ± 1 ℃ for 18-24h.
Procedures:
1. Prepared sample solution according to
national standards, SN standard, FDA standards or other methods.
2. Transfer 1mL sample onto chromogenic
medium cooling down to 45-50 ℃, mix thoroughly or coating on plate;
3. Culture at 36 ± 1 ℃ for 18-24h, select the plate which have 30
to 200 typical colonies of E. coli, total count of the colonies on the plate,
typically E. coli colonies blue-green, others yellow or colorless colonies.
4. For undermined colonies, streak them on
nutrient agar plates, culture at 36 ± 1 ℃ for 12-16h, select single colony to do BGLB
Fermentation tests.
Quality Control:
1. Appearance:
The dehydrate powder medium is homogeneous,
having good flowability, milky white, prepared medium is colorless solid.
2. Microbial sensitivity test:
Incubated at 36 ± 1 ℃ 18-24h:
QC Strains
|
Strains No.
|
Growth
|
Colonies color
|
Listeria
monocytogenes
|
ATCC27853
|
-/+
|
colorless
|
Staphylococcus
aureus
|
ATCC25923
|
-/+
|
colorless
|
E. coli
|
ATCC25922
|
+++
|
blue
|
salmonella
|
ATCC14028
|
+++
|
colorless
|
Enterobacter
aerogenes
|
ATCC35029
|
+++
|
blue
|
Enterobacter
cloacae
|
ATCC23355
|
+++
|
blue
|
Freundii Citrobacter
|
ATCC43864
|
+++
|
blue
|
Klebsiella pneumoniae
|
ATCC10031
|
+++
|
blue
|
Methods limitations:
The medium identification of Coliforms may
occur false-positive in a few cases, but no false-negative, sometimes it needs
to do other complementary tests
Typical features:
Coliforms typical colonies are blue-green,
coliforms colonies are colorless, other bacteria are colorless or yellow
colonies.
Storage:
The prepared plates must be used
immediately, 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. FRAMPTON, E.W., RESTAINO, L. a. BLASZKO,
L.: evalsuation of ß-glucuronidase substrate in a 24 hour direct plating method
for Escherichia coli. J. Food
Protection, 51; 402-404 (1988).
2. KILIAN, M. a. BüLOW, P.: Rapid diagnosis
of Enterobacteriaceae. I. Detection of bacterial glycosidases. Acta Pathol.
Microbiol. Scand. Sect. B 84; 245-251 (1976).
3. LE MINOR, L. a. HAMIDA, F. BEN:
Advantages de la recherche de la ßgalactosidase sur celle de la fermentation du
lactose en milieu complexe dans le diagnostic bactériologique, en particulier des Enterobacteriaceae. Ann.
Inst. Pasteur (Paris), 102; 267-277 (1962).