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V4 – 12/08/11

YGC/Agar 355-5489
(Yeast - Glucose - Chloramphenicol) 356-4104

DEFINITION EQUIPMENT REQUIRED (NOT SUPPLIED)


Medium used for the enumeration of yeasts and (non-exhaustive)
molds in the analysis of food products. • Scales
• Sterile weighing bags
STANDARDS • Grinder
FOOD MICROBIOLOGY • Hotplate
• NF ISO 7698 (August 1991): Cereals, • Mixer-homogenizer
leguminous products and derivative products - • 100 ml Pyrex bottles with autoclave-proof
Enumeration of bacteria, yeasts and moulds stoppers
(IC: V03-763). • Sterile Petri dishes (Ø = 90 mm)
• Sterile Pasteur pipettes (0.1 ml, 1 ml, etc)
• NF ISO 7954 (August 1988): Microbiology - • Sterile spreaders
General guidelines for the enumeration of • Water-bath precise to ± 1°C
yeasts and molds. Colony-count technique at • Thermostatically-controlled incubator or
25°C (IC: V08-022). incubation room, precise to ± 1°C
• Autoclave
• FIL 94B (1991): Milk and milk products - • All usual laboratory equipment.
Enumeration of yeasts and molds by colony
count at 25°C. PREPARATION OF DEHYDRATED MEDIUM
Always shake before use.
PRINCIPLE Dissolve 41.1 g of powder in 1 liter of distilled
The nutrient substances provided by yeast water. Wait for 5 minutes, then mix until a
extract and the glucose used as energy source homogenous suspension is obtained. Heat
favor the growth of yeasts and moulds. The gently swirling frequently, then bring to boiling
presence of chloramphenicol, a thermostabile point until completely dissolved.
broad-spectrum antibiotic, inhibits the growth of Dispense 100 ml per bottle, then sterilize in
contaminant bacteria. autoclave at 121°C (± 1°C) for 15 minutes.
PRESENTATION Reconstitution ratio: 41.1 g/l.
• Ready-to-use 500 g of powder makes 12.1 liters of
100 ml x 6 bottles code 355-5489 medium.
• Dehydrated PROTOCOL
500 g code 356-4104 N.B.: In cases where heavy contamination by
Gram negative bacteria is suspected
STORAGE (particularly in meat or raw fish), it is
• Ready-to-use: + 2°C to 25°C recommended that a solution of Gentamicin be
• Dehydrated: + 15°C to 25°C, in carefully- added to the Y.G.C. medium (final
sealed bottles in a cool, dry place. concentration = 100 mg/l), previously cooled to
• Expiration date and batch number are 44°C- 47°C.
shown on the package.
• Inoculation
THEORETICAL FORMULA Surface
Yeast extract 5g - Spread 0.1 ml of test sample or 0.1 ml of the
Glucose 20 g stock solution (other products) and/or 0.1 ml
Chloramphenicol 0.1 g of its decimal dilutions on the surface of the
Agar 16 g “dried” agar.
Disilled water 1,000 ml - Leave to dry.
Final pH (25°C) = 6.6 ± 0.2
Depth
OTHER PRODUCTS REQUIRED - Transfer 1 ml of sample or 1 ml of stock
(NOT SUPPLIED) suspension (other products) and/or 1 ml of its
• Distilled water decimal dilutions.
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V4 – 12/08/11
YGC/Agar
(Yeast - Glucose - Chloramphenicol)

- Pour 12 to 15 ml of medium, melted and Documentation relative to the production and


cooled to 44°C- 47°C. control of each batch is kept on file.
- Homogenize and leave to cool on a cool, level
surface. PERFORMANCES / QUALITY CONTROL OF
THE TEST
• Incubation The growth performances of the media are
Turn over the plates and incubate at 25°C verified with the following strains:
(± 1°C) for 3 to 5 days, depending on the
standards. Result after 3-5
STRAINS
days at 25°C
READING AND INTERPRETATION
• Colony count (UFC) Productivity
Enumerate the colonies on each plate after 3, 4
Candida albicans
and 5 days, according to the standards. ATCC 10231
PR ≥ 0.5

N.B.: Aspergillus niger


- If, after the last day of incubation, it is difficult PR ≥ 0.5
ATCC 16404
to count highly isolated colonies, use the
counts obtained on the previous day(s). Penicillium cyclopium
PR ≥ 0.5
- If necessary, distinguish colonies of yeasts ATCC 16025
and molds from colonies of bacteria by
Saccharomyces cerevisiae
microscope examination (morphological PR ≥ 0.5
ATCC 9763
characteristics).
- In general, select plates containing fewer than Selectivity
150 colonies (minimum 15).
- Depending on the calculation method, plates Escherichia coli
No growth
containing fewer than 15 colonies or no ATCC 25922
colonies can be selected (small number
Bacillus subtilis
estimation). No growth
ATCC 6633
• Expression of results/Calculations
* PR = Total colony count obtained on 2 plates of
For the calculation method, refer to standard YGC/total colony count on 2 plates of Sabouraud
NF ISO 7218 and the specific standard. agar.

PRECAUTIONS
- Avoid prolonged heating during melting. KEY WORDS
- The medium may look frothy after gelification YGC / Yeasts / Molds / Food products /
in bottles. It nevertheless keeps all its Enumeration / Chloramphenicol / Medium.
qualities as through its appearance is
changed by melting and shaking. BIBLIOGRAPHY
- The time lapse between the end of • SAINCLIVER M. - ROBLOT A.M. (1966):
preparation of the stock solution (or the 10-1 Choix d’un milieu de culture pour le
dilution in the case of a solid product) and the dénombrement des levures et moisissures
moment when the dilutions come into contact dans le beurre. Annales de l’Institut Pasteur.
with the culture medium must not exceed 15 17: 181.
minutes.
- The Petri dishes should be handled with care • MOSSEL D.A.A. - VISSER M. - MENGERINK
in order to avoid the dispersion of mold W.H.J. (1962): A comparison of media for the
spores. enumeration of molds and yeasts in food and
- Comply with Good Laboratory Practice. beverages. 2ab. Pract. 11:109-112.

QUALITY CONTROL OF MANUFACTURER


Every product manufactured and marketed by
Bio-Rad is subject to a quality-assurance
procedure at all stages, from the reception of
raw materials to the marketing of the end-
product. Each batch of finished product
undergoes quality control and is marketed only
if it satisfies the acceptability criteria.

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