E.Coli & Coliforms Chromogenic Medium is used for fast, accurate simultaneous detection of E. coli and Coliform bacteria, incubated for 24 hours, E. coli was blue-green - purple, coliform is red.
Ingredients (g/L):
Special
Nutrients
|
39.0
|
Mixed Chromogenic
reagent
|
0.2
|
Agar
|
13.0
|
Final pH 7.0-7.2
at 25℃
|
*Adjusted and/or supplemented as required
to meet performance criteria.
Directions:
Suspend 52.2g of the medium in 1L of diatilled 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 200
to 300 typical colonies of E. coli and coliforms, total count of the colonies
on the plate, typically E. coli colonies blue to purple, coliforms are pink, 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 full
set of E.Coli biochemical 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
|
-/+
|
blue
|
E. coli
|
ATCC25922
|
+++
|
colorless
|
salmonella
|
ATCC14028
|
+++
|
pink
|
Enterobacter
aerogenes
|
ATCC35029
|
+++
|
pink
|
Freundii Citrobacter
|
ATCC43864
|
+++
|
pink
|
Klebsiella
pneumoniae
|
ATCC10031
|
+++
|
pink
|
Methods limitations:
The medium identification of E.Coli./Coliforms
may occur false-positive in a few cases, but no false-negative, sometimes it
needs to do other complementary tests.
Typical features:
E.Coli typical colonies are blue to purple,
coliforms colonies are pink, 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.: Evaluation 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).