HiCrome™ Bifidobacterium Agar

Intended use

Recommended for the differentiation of Bifidobacterium and Lactobacillus species from clinical and non-clinical samples.

Composition**

Final pH ( at 25°C): 7.2±0.2

**Formula adjusted, standardized to suit performance parameters

# - Equivalent to Milk powder

Directions

Suspend 59.48 grams in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

The genus Bifidobacterium is the third most numerous bacterial populations found in the human intestine after Bacteroides and Eubacterium. It is an anaerobic bacteria that makes up the gut microbial flora. It resides in the colon and have health benefits for their hosts. Bifidobacteria are also associated with lower incidences of allergies (1,2). Bifidobacterium Agar is used for the cultivation and maintenance of Bifidobacterium species (3).

Peptone special provides nitrogenous and carbonaceous compounds, long chain amino acids, vitamins and other essential growth nutrients. Sodium chloride maintains osmotic balance. M-protein powder aids in detecting casein hydrolysis activity which is exhibited by Bifidobacterioum breve. A halo zone is observed around the colony in case of casein hydrolysis. The indicator system in the chromogenic mixture helps in distinguishing between Lactobacillus and Bifidobacterium species. Lactobacillus species usually produce green colonies with opaque zone. Bifidobacterium infantis produces dark blue to bluish green colonies. Agar serves as a solidifying agent.

Type of specimen

Clinical samples - Faeces, gut swab, etc.; Food and dairy samples.

Specimen Collection and Handling

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (4,5). For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (6,7,8). After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

In Vitro diagnostic Use. For professional use only. 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 clinical specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations

1. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
2. Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user’s unique requirement.
3. Slight colour variation may be observed depending upon the utilization of the substrate by the organism.
4. Bifidobacterium species are strict anaerobes, hence condition must be appropriately maintained.

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 Cream to yellow homogeneous free flowing powder

Gelling Firm, comparable with 1.6% Agar gel

Colour and Clarity of prepared medium Reddish orange coloured clear to slightly opalescent gel forms in Petri plates

Reaction Reaction of 5.95% w/v aqueous solution at 25°C. pH :7.2±0.2

Cultural Response Cultural characteristics observed after an incubation at 35-37°C for 48 hours under anaerobic conditions.

Storage and Shelf Life

Store between 15-30°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 (4,5).

Reference

1. Björkstén B., Sepp E., Julge K., Voor T., and Mikelsaar M., 2001, J. Allergy Clin. Microbiol., Volume 108, Issue 4, 516-520.
2. Guarner F., and Malagelada J. R., 2003, The Lancet, Vol. 361, Issue 9356, 8 February 2003, 512-519.
3. Atlas R. M. 2004, 3rd Edi. Handbook of Microbiological Media, Parks, L. C. (Ed.), CRC Press, Boca Raton.
4. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
5. 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.
6. American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., Washington D.C.
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.