Karmali Campylobacter Agar Base

Intended use

Karmali Campylobacter Agar is a blood free medium recommended for selective isolation and cultivation of thermotolerant Campylobacter species from food, animal feeds and clinical samples.

Composition**

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 22.5 grams in 490 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. Aseptically add rehydrated contents of 1 vial of Campylobacter Selective Supplement w/ Hemin (Karmali) (FD132) or Campylobacter Selective Supplement w/ Hemin (Karmali), Modified (FD167). Alternatively, add aseptically rehydrated contents of one vial of Campylobacter Selective Supplement, Karmali (FD078) or Campylobacter Selective Supplement (Karmali), Modified (FD178) and 5 ml of Hemin solution (16 mg/5 ml). Mix well and pour into sterile Petri plates.

Principle And Interpretation

Campylobacter are carried in the intestinal tract of animals and therefore, contaminate foods of animals origin. Campylobacter jejuni is recognized as a leading cause of acute bacterial gastroenteritis in humans, and eating foods of animal origin has been associated with many of these illnesses (9). Campylobacter jejuni and Campylobacter coli are the most common Campylobacter species associated with diarrheal illness and are clinically indistinguishable (8). Karmali Campylobacter Agar Base, recommended for the selective isolation and cultivation of Campylobacter species, is a modification of the original formulation of Karmali et al (7). Selectivity of the medium is achieved by the addition of selective supplement. Campylobacter Selective Supplement with Hemin (Karmali) (FD132) or Campylobacter Selective Supplement with Hemin (Karmali), Modified (FD167) has hemin, as part of the supplement whereas, while using Campylobacter Selective Supplement, Karmali (FD078) or Campylobacter Selective Supplement (Karmali), Modified (FD178), hemin has to be added separately. Karmali Campylobacter Agar Base is also recommended by the ISO Committee (4).

Peptone special, cornstarch and hemin, serve as sources of essential nutrients required for bacterial metabolism. Presence of charcoal in the medium helps to neutralize the toxic metabolic products formed in the medium. Sodium pyruvate (present in Supplement) (2) enhances, the aerotolerance of microaerophilic Campylobacter by quenching the toxic forms of oxygen (3). The antibiotics included in the selective supplement are Vancomycin, Ammphotericin B, Cycloheximide and Cefoperazone. Vancomycin suppresses gram-positive organisms while Amphotericin B/ Cycloheximide inhibits the fungal flora. Cefoperazone has inhibitory action on gram-negative organisms other than Campylobacter. The inoculated plates are incubated in an atmosphere consisting of approximately 5-6% O₂, 10% CO₂ and 84-85% N₂ at 42°C.

Type of specimen

Clinical samples- faeces; Food and dairy samples

Specimen Collection and Handling

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (5,6).

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (1,9,10).

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.

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 Grey to black homogeneous free flowing powder

Gelling Firm, comparable with 1.2% Agar gel.

Colour and Clarity of prepared medium Black coloured, opalescent gel forms in Petri plates

Reaction Reaction of 4.5% w/v aqueous solution at 25°C. pH : 7.4±0.2

pH 7.20-7.60

Cultural Response

Cultural characteristics observed with added Hemin solution and Campylobacter Selective Supplement (Karmali), Modified (FD178)/ Campylobacter Selective Supplement, Karmali (FD078) or Campylobacter Selective Supplement w/ Hemin (Karmali), Modified (FD167)/ Campylobacter Selective Supplement w/Hemin(Karmali)(FD132) after an incubation at 42°C for 42-48 hours.

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. George H. A., Hoffman P. S. and Krieg N. R., 1978, J. Clin. Microbiol., 8:3
3. Hoffman P. S. et al, 1979, Can. J. Microbiol., 25:
4. International Organization for Standardization (ISO), 1995, Draft ISO/DIS 1027
5. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Editio
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. Karmali M. A., Simor A. E., Roscoe M., Fleming P. C., Smith S. S. and Lane J., 1986, J. Clin. Microbiol., 23:456-459.
8. Murray P. R., Baron E. H., Pfaller M. A., Tenover F. C. and Yolken R. H., (Ed.), 1995, Manual of Clinical Microbiology, 6th Ed., American Society for Microbiology, Washington, D.C.
9. 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.
10. Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed., APHA Inc., Washington, D.C.