Description: used for
the rapid detection of Bacillus cereus in food, water, dairy products and meat
products. Improved chromogenic reagents and additives have enhanced specificity
and sensitivity significantly. The colonies of Bacillus cereus are blue-green
and have an opaque ring around, other bacteria are blue-green, yellow or
colorless, Gram-negative bacteria are inhibited.
Ingredients (g / L)
Special
Nutrients
|
20.0
|
Chromogenic
reagent
|
0.5
|
Bacteriostatic
agent
|
4.0
|
Agar
|
15.0
|
Finnal pH 7.0±0.2 at 25 ℃
|
*Adjusted and/or supplemented as required
to meet performance criteria
Directions:
Suspend basal medium 7.95g of 180mL of
distilled water; add Bacillus cereus additive 0.6g and 20ml of sterile water, put
into some glass balls, constant vibrating for 5-10min to emulsify uniformly,
autoclave the above two at 121 ℃ for 15min. When cooling to about 45-50 ℃,add the Bacillus cereus additive into basic medium,
and mix thoroughly, pour into sterile petri dishes. Make the medium thickness is
about 1-2mm, so advantageous to observe opaque ring.
Procedures:
1. Prepared sample solution according to
national standards, SN standard, FDA standards or other methods.
2. Enrichment culture for the samples
solution at 30 ± 1 ℃ for 18- 24 hours.
Transfer
25g sample solution into 225mL GN Broth or Shigiella Broth, enrichment culture
for 18- 24hours at 36 ± 1 ℃;
3. Take enrichment broth streak to inoculate
Bacillus Cereus Chromogenic Medium (Generation Ⅲ), culture
at 36 ± 1 ℃ for 18- 24 hours, the typical colonies of Bacillus cereus
are blue-green and have an opaque ring around, if not so in 24 hours, culture
for 48 hours.
4. For
undetermined colonies, streak on nutrient agar plate, culture for 18- 24hours
at 36 ± 1 ℃, select single colony to do full set of 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
|
Bacillus
Cereus
|
CMCC63303
|
+++
|
Dark
blue, an opaque ring around colony
|
Bacillus
Cereus
|
ATCC11778
|
+++
|
Blue-green,
large and flat and an opaque ring around colony
|
Bacillus
subtilis
|
CMCC63501
|
++
|
White
colonies, no opaque ring around
|
Staphylococcus
aureus
|
ATCC25923
|
-/+
|
white
|
E. coli
|
ATCC25922
|
-
|
white
|
Methods limitations:
The identification of Bacillus cereus
medium, sometimes there may be a false positive, it needs be done other
complementary tests. The color of Bacillus thuringiensis on chromogenic medium
is the same as Bacillus cereus, but rarely inspect in food
Typical features:
The typical colonies of Bacillus cereus are
blue-green and have an opaque ring around
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. P.R. Mortimer, G. McCann, Lancet, 1043
(1974)
2. E. Bouza, S. Grant, C. Jordan, et. al,
Arch. Ophthamol., 97, 488 (1979)
3. K. Wohlgemuth, C.A.
Kirkbride, E.J. Bicknell, R.P. Ellis, Am. Vet. Met, Ass., 161, 1691 (1972)