WL Nutrient Broth M050
Intended Use:
Recommended for cultivation of yeasts, moulds and bacteria encountered in brewing and industrial fermentation processes.
Composition**
Ingredients Gms / Litre
Tryptone 5.000
Yeast extract 4.000
Dextrose (Glucose) 50.000
Potassium dihydrogen phosphate 0.550
Potassium chloride 0.425
Calcium chloride anhydrous 0.125
Magnesium sulphate 0.125
Ferric chloride 0.0025
Manganese sulphate 0.0025
Bromo cresol green 0.022
Final pH ( at 25°C) 5.5±0.2
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 60.25 grams in 1000 ml purified / distilled water. Heat if necessary, to dissolve the medium completely.Dispense in
tubes or flasks or as desired Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. If desired
to obtain a pH of 6.5, add 1% solution of sodium bicarbonate before sterilization.
Principle And Interpretation
WL (Wallerstein Laboratory) media are formulated as described by Green and Gray for the examination of materials
encountered in brewing and for industrial fermentations containing mixed flora of yeast and bacteria (1, 2). Bakers yeast
counts can be carried out in this medium at a pH 5.5. By adjusting the pH to 6.5, the medium can be used for obtaining
counts of Baker and distillers yeast (5). If desired Durham's tubes can be added to WL Nutrient Broth to study fermentation
reactions.
Yeast extract serves as a source of trace elements, vitamins and amino acids. Tryptone is used as a source of nitrogen, amino
acids and carbon. Dextrose is the source of carbohydrate. Buffering of the medium is done by monopotassium phosphate.
Potassium chloride, calcium chloride and ferric chloride are essential ions that help to maintain the osmotic balance.
Magnesium sulphate and manganese sulphate are sources of divalent cations. Bromo cresol green is a pH indicator.
Type of specimen
Microorganisms in brewery
Specimen Collection and Handling:
For fermentation samples follow appropriate techniques for handling specimens as per established guidelines (1,2).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions :
Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/ face
protection. Follow good microbiological lab practices while handling specimens and culture. Standard
precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be
referred in individual safety data sheets.
Limitations :
1. Further biochemical identification of organisms is required for confirmation.
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data
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
Ligth yellow to light green homogeneous free flowing powder
Colour and Clarity of prepared medium
Bluish green coloured very slightly opalescent solution in tubes.
Reaction
Reaction of 6.02% w/v aqueous solution at 25°C. pH : 5.5±0.2
pH
5.30-5.70
Cultural Response
Cultural characteristics observed in tubes containing inverted Durham's tubes after an incubation at 35-37°C for 40-48 hours
for bacteria and at 30-32°C upto 5 days for yeast.
Organism Inoculum Growth Acid Gas
(CFU)
Escherichia coli ATCC 50-100 fair-good positive positive
25922 (00013*) reaction, yellow
colour
Lactobacillus fermentum 50-100 fair-good positive slightly positive
ATCC 9338 reaction, yellow
colour
Saccharomyces cerevisiae 50-100 good positive positive
ATCC 9763 (00058*) reaction, yellow
colour
Key : *Corresponding WDCM numbers.
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°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 sample must
be decontaminated and disposed of in accordance with current laboratory techniques (3,4).
Reference
1. Green S. R. and Gray P. P., 1950, Wallerstein Lab. Commun., 12:43
2. Green S. R. and Gray P. P., 1950, Wallerstein Lab. Commun., 13:357
3. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
4. 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.
5. MacFaddin J. F., 1985, Media for Isolation-Cultivation- Identification- Maintenance of Medical Bacteria, Vol.1,Williams
& Wilkins, Baltimore, Md.
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 or therapeutic use but
for laboratory,diagnostic, 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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