Carbohydrate Consumption Broth Base M1264
Intended Use:
Recommended for cultivation and differentiation of Listeria species.
Composition**
Ingredients Gms / Litre
Proteose peptone 10.000
Sodium chloride 5.000
HM peptone B # 1.000
Bromocresol purple 0.100
Final pH ( at 25°C) 6.8±0.2
**Formula adjusted, standardized to suit performance parameters
# - Equivalent to Beef extract
Directions
Suspend 16.1 grams in 990 ml purified / distilled water. Heat if necessary to dissolve the medium completely. Dispense
into tubes containing inverted Durhams tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Aseptically add 10 ml separately sterilized carbohydrate solution to give a final concentration of 0.5%. Mix well.
Principle And Interpretation
Carbohydrate Consumption Broth is used for the cultivation and differentiation of Listeria species and formulated as per
Atlas (2). It is also recommended by FDA (3) and ISO (4) with a slight difference in the concentration of bromocresol
purple. Differentiation is based on fermentation of glucose, xylose, rhamnose, ribose, a-methyl-D-mannoside and mannitol.
Proteose peptone and HM peptone B in the medium provide carbon and nitrogen compounds including essential amino
acids, vitamins and trace ingredients for bacterial metabolism. Bromocresol purple is the pH indicator, which indicates acid
production by turning yellow in colour.
Carbohydrate utilization test: Inoculate each kind of carbohydrate fermentation broth with one loopful of inoculum. Incubate
for 7 days at 37°C. Observe daily for acid induced colour change and gas formation. Sometimes weak positive reactions may
occur after 48 hours of incubation (3).
Type of specimen
Isolated Microorganism from food samples.
Specimen Collection and Handling:
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (1,7,8).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Limitations :
NA
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at
recommended temperature.
Quality Control
Appearance
Light yellow to beige homogeneous free flowing powder
Colour and Clarity of prepared medium
Purple coloured, clear solution without any precipitate
Reaction
Reaction of 1.61% w/v aqueous solution at 25°C. pH : 6.8±0.2
pH
6.60-7.00
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
Organism Inoculum Growth w/o w/o w/ rhamnose w/ rhamnose
(CFU) carbohydrate carbohydrate (acid) (gas)
acid gas
Escherichia coli ATCC 50-100 good-luxuriant negative negative positive positive
25922 (00013*) reaction, no reaction reaction, yellow reaction
colour change colour
Listeria monocytogenes 50-100 good-luxuriant negative negative positive negative
subsp. serovar 1 ATCC reaction, no reaction reaction, yellow reaction
19111 (00020*) colour change colour
Listeria monocytogenes 50-100 good-luxuriant negative negative positive negative
ATCC 19112 reaction, no reaction reaction, yellow reaction
colour change colour
Listeria monocytogenes 50-100 good-luxuriant negative negative positive negative
ATCC 19117 reaction, no reaction reaction, yellow reaction
colour change colour
Staphylococcus aureus 50-100 good-luxuriant negative negative negative negative
ATCC 25923 (00034*) reaction, no reaction reaction, no reaction
colour change colour change
Listeria monocytogenes 50-100 good-luxuriant negative negative positive negative
ATCC 19118 reaction, no reaction reaction, yellow reaction
colour change colour
Key : *Corresponding WDCM numbers.
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on
the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent
lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump
formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition Seal the
container tightly after use.
Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product.
Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with
clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (5,6).
Reference
1. American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed.,
Washington D.C.
2. Atlas R. M., 2004, Handbook of Microbiological Media, 3rd Edition, CRC Press, Washington D. C.
3. FDA Bacteriological Analytical Manual, 2005, 18th Ed., AOAC, Washington, D.C.
4. International Organization for Standardization (ISO), 1993, Draft ISO/DIS 10560.
5. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
6. Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W.
(2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.
7. Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of
Foods, 5th Ed., American Public Health Association, Washington, D.C.
8. Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th
Ed., APHA Inc., Washington, D.C.
Revision : 03 / 2019
Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in this
and other related HiMedia™ publications. The information contained in this publication is based on our research and development work
and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to specifications
and information related to the products at any time. Products are not intended for human or animal diagnostic or therapeutic use but for
laboratory, research or further manufacturing use only, unless otherwise specified. Statements contained herein should not be considered
as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.
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