TIME SCALE AND SCHEMATIC REPRESENTATION OF MICROBIOLOGICAL ANALYSIS OF FOOD and BEVERAGE SAMPLES AS FSSR, 2011

Food and Beverage

Sample

3 4 5 6
Test 1 2
No
Coliforms count/ ml or g Escherichia coli / ml or ml
Total Plate count/ Aerobic Yeast and Mould count Staphylococcus aureus/ g Staphyloccus aureus /25g
Microbial count/ml or g / ml or g
10g/ml of Sample + 90ml of 0.1% Sterile Peptone Water
(dilution 10-1)/ Homogenize. Decimal dilution up to 10-3 Aseptically weigh 50 gm
25g sample + 225ml of
food sample into the sterile
10g/ml of Sample + 90ml of 10g/ml of Sample + 90ml of Butterfields Buffered
blender jar. Add 450mL of
0.1% Sterile Peptone Water 0.1% Sterile Peptone Water Pour Plating: Pipette1ml of dilution into appropriately phosphate. Homogenize
diluent (1:10) and
(dilution 10-1)/ Homogenize (dilution 10-1)/ Homogenize marked petri dish and pour 10ml of molten Violet Red Bile 2min at high speed.
homogenize 2 min at high
agar (48oC). Overlay with 3-5ml VRBA. Perform pour plating speed.
in duplicates. Allow plates to set at room temperature.
Make decimal dilutions with 10g/ml of Sample + 90ml
Pour Plating: Pipette1ml of Incubation: Incubate Plates inverted at 37oC /-24hrs.
1ml food homogenate of dilution broth
food homogenate into
using 0.1% Peptone Water
appropriately marked petri 10g/ml of Sample + 90ml of
up to 10-3 dilution Count all colonies that are purple red in color, 0.5 mm in
dish and pour 10ml of dilution broth
molten acidified Potato diameter or larger and are surrounded by a zone of
Dextrose agar (45oC) precipitated bile acids. Optimally the count should be
Pour Plating: Pipette1ml of between 15-150 (Plate diameter of 90 to 95 mm). 10g/ml of Sample + 90ml
dilution into appropriately 10g/ml of Sample + 90ml of of dilution broth
marked petri dish and pour dilution broth
10ml of molten Standard Calculation: Inoculate 5purple red
Perform pour plating in colonies from VRBA into
Plate Count agar (45oC)
duplicates. Allow plates to N= C/ [N1 + 0.1N2]D
Report xcfu/g orml. 2ml of LST broth, Incubate 10g/ml of Sample + 90ml of 10g/ml of Sample + 90ml
set at room temperature for 2-4 has at 37oC. dilution broth of dilution broth
Perform pour plating in
duplicates. Allow plates to Streak one plate EMB agar to obtain discrete colonies.
set at room temperature Incubation: Incubate plates Incubate plates inverted at 37oC/ 24hrs.
inverted at 25oC for 24hrs.
10g/ml of Sample + 90ml of 10g/ml of Sample + 90ml
Examine plates for typical nucleated dark centered colonies dilution broth of dilution broth
Incubation: Incubate Plates with orwithout sheen.. Transfer growth to PCA slant and
inverted at 37oC /-24hrs Observe for typical yeast
incubate at 37oC/24hrs.
and mould colonies. Do
not open plate containing
Transfer growth from PCA slants to the following media for 10g/ml of Sample + 90ml of 10g/ml of Sample + 90ml
Observe for discrete mould colonies. Re-
biochemical test. dilution broth of dilution broth
colonies. Count the colonies incubate plates up to 5-7
using Colony Counter days and look for growth
before reporting -ve Tryptone Broth: Incubate at 37°C/ 24hr and test for indole.

MR-VP Medium: Incubate 48hrs at 37°C. Aseptically transfer

Calculation: Count colonies and report 1ml of culture to a tube and perform the VP test. Incubate
N= C/ [N1 + 0.1N2]D count after taking account the remaining culture an additional 48h and test for methyl 10g/ml of Sample + 90ml
Report xcfu/g orml of dil. factor x cfu/ml or g red reaction. of dilution broth

Koser citrate broth: Inoculate koser citrate broth and Incubate

96 hours at 37+0.5°C and record growth.

Report as Eshchericahiia colil - x cfu,g or ml