M-Kleb Agar Base

Intended Use

Recommended for selective isolation and differentiation of Klebsiella from water and other sources.

Composition**

Final pH ( at 25°C): 7.4±0.2

**Formula adjusted, standardized to suit performance parameters

# Equivalent to Beef Extract

Directions

Suspend 36.22 grams in 980 ml 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. Add 20ml of 95% Ethyl alcohol to 980ml of media and aseptically add rehydrated contents of two vials of Klebsiella Selective Supplement (FD225). Mix well and pour into sterile Petri plates.

Principle And Interpretation

m-Kleb Agar Base is recommended for isolation and differentiation of Klebsiella species by membrane filtration method Klebsiella pneumoniae strains are widely distributed in the environment and contribute to biochemical and geochemical process (1). K.pneumoniae bacteria may be opportunistic pathogens that can give rise to bacteremia, pneumonia, urinary tract, and several other types of human infection. It also proves to be the source of lung infections that generally occur in patients with debilitating conditions such as alcoholism, diabetes mellitus, and chronic obstructive pulmonary disease (2). K.pneumoniae are also excreted in the faeces of many healthy humans and animals, and they are readily detected in sewage polluted waters (3).

K.pneumoniae produces a deep blue to bluish green coloured colony thereby aiding in the easy detection of the organisms. Most of the frequently encountered gram-negative faecal contaminants are inhibited on this media using a selective supplement.

Proteose peptone, HM peptone B provides nitrogenous and carbonaceous compounds, long chain amino acids, vitamins and other essential nutrients required for the growth of the organism. Inositol us the fermentable carbohydrate. Neutral red and aniline blue are the pH indicators. Sodium chloride maintains the osmotic equilibrium of the medium. Sodium lauryl sulphate (SLS) inhibit most of the accompanying flora. Addition of the selective supplement further increases the selectivity of the medium.

Type of specimen

Water samples

Specimen Collection and Handling

For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(4)

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

This medium is selective medium, however confirmatory test has to be carried out.

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 pink homogeneous free flowing powder

Gelling: Firm, comparable with 1.5% Agar gel

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

Reaction: Reaction of 3.62% w/v aqueous solution at 25°C. pH : 7.4±0.2

pH: 7.20-7.60

Cultural Response: Cultural characteristics observed on membrane filter with added Klebsiella Selective Supplement (FD225) after an incubation at 35-37°C for 18-24 hours.

Key : * Corresponding WDCM Numbers

# Formerly known as Klebsiella aerogenes

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 inorder 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. Use before expiry date on the label.

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 (5,6).

Reference

1. Krieg, N. R., and J. G. Holt, (Eds.), 1984, Bergeys Manual of Systematic Bacteriology, Vol. 1, p. 408 - 516. The Williams and Wilkins Co., Baltimore, MD.
2. Wyngaarden J. B., Smith L. H., (Eds.), Cecil Text book of Medicine,16th Ed, pp 1430 -1432, Philadelphia, W. B. Saunders, 1982.
3. Standard methods, For the examination of water and wastewater, 22nd edition, Eugene W. Rice, Rodger B. Baird, Andrew D. Eaton, Lenore S. Clesceri.
4. Greenberg A. E., Clesceri L. S. and Eaton A. D., (Eds.), 2005, Standard Methods for the Examination of Water and Wastewater, 21st ed., APHA, Washington, D.C.
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.