Dubos Oleic HiVeg™ Broth Base

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MV839
Recommended for cultivation of Mycobacteria.


Intended Use

Recommended for cultivation of Mycobacterium tuberculosis and for determining its sensitivity to chemo-therapeutic agent.

Composition

Ingredients g/L
HiVeg™ hydrolysate 0.500
L-Asparagine 1.000
Potassium dihydrogen phosphate 1.000
Disodium hydrogen phosphate 2.500
Ferric ammonium citrate 0.050
Magnesium sulphate 0.010
Calcium chloride anhydrous 0.0005
Zinc sulphate 0.0001
Copper sulphate 0.0001
Final pH (at 25°C) 6.6±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 1.0 gram in 180 ml purified/distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool and aseptically add 1 vial (20 ml) of sterile Oleic Albumin Supplement (FD020) and 5,000 to 10,000 units of Penicillin. Mix well and dispense in sterile tubes or flasks as desired.

Principle And Interpretation

Tuberculosis remains a major global public health problem worldwide. Mycobacterium tuberculosis, the causative agent of tuberculosis in man, is carried in airborne particles known as droplet nuclei that are generated when patients with pulmonary tuberculosis cough. Infections occur when a susceptible person inhales the droplet nuclei containing the bacterium (1). Dubos Oleic Broth Base is recommended by Dubos and Middlebrook (1) for the primary isolation and subsequent cultivation of the tubercle bacilli. On comparative studies of various media, Dubos Oleic Agar Base was found to be superior to other media for the primary isolation of Mycobacteria (2,3). Mycobacteria grow very rapidly when inoculated in a broth media and therefore preliminary culture of all the test samples in a broth media is recommended. Dubos Oleic HiVeg™ Broth Base is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. Dubos Oleic HiVeg™ Broth Base contain HiVeg™ hydrolysate and L-aspargine as sources of nitrogen. The phosphates (together with calcium chloride) buffers the media as well as serve as sources of phosphates. Magnesium sulphate, zinc sulphate, copper sulphate and ferric ammonium citrate provide trace metals and sulphates. Standard procedures for the isolation of Mycobacteria from test samples should be followed (4). The specimen should be appropriately decontaminated before culturing as per standard methods (4,5,6,7). Maximum care should be taken while handling Mycobacterial cultures, as they are highly infectious.

Type of specimen

Clinical samples : Sputum

Specimen Collection and Handling

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:

  • Proper aerobic conditions and increased CO2 tension if not provided during incubation, may lead to erroneous result.

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
Off-white to beige homogeneous free flowing powder

Colour and Clarity of prepared medium
Light amber coloured, clear to slightly opalescentsolution with a fine precipitate.

Reaction
Reaction of medium (0.5% w/v aqueous solution containing 0.1% FD020)at 25°C. pH: 6.6±0.2

pH
6.40-6.80

Cultural Response
Cultural characteristics observed with added Oleic Albumin Supplement (FD020) and 5000-10,000 units of Penicillin, after an incubation at 35-37°C for 2-6 weeks.

Organism Growth
Mycobacterium avium (25291) luxuriant
Mycobacterium gordonae ATCC 14470 luxuriant
Mycobacterium kansasii ATCC 12478 luxuriant
Mycobacterium smegmatis ATCC 14468 luxuriant
Mycobacterium tuberculosis H37RV (25618) luxuriant

Storage and Shelf Life

Store below 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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5,6,7,8).

More Information
Product Name Dubos Oleic HiVeg™ Broth Base
SKU MV839
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Dubos R. J., and Middlebrook G., 1947, Am. Rev. Tuberc., 56:334.2.Roberts A. H., Wallace R. J. and Erlich P., 1950, Am. Rev. Tuberc., 61:563.3.Byham, 1950, Am. J. Clin. Pathol., 20:6784.Kent and Kubica, 1985, Public Health Mycobacteriology : A Guide For the Level III Laboratory, USDHHS, Center forDisease Control, Atlanta c.a.5.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of Clinical Microbiology,8th Ed., American Society for Microbiology, Washington, D.C.6.Isenberg (Ed.), 1994, Clinical Microbiology Procedures Handbook, Suppl. 1., American Society for Microbiology,Washington, D.C.7.Forbes B. A., Sahm A. S., and Weissfeld D. F., Bailey & Scotts Diagnostic Microbiology, 10th Ed., 1998, Mosby, Inc.,St. Louis, Mo.8.. 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
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