Here are some tips:

1. Enrichment in broth with 5-10% defibrinated blood (sheep or horse) will increase the chances of recovering C. jejuni.

2. Consider using Campy selective supplement (e.g., from Himedia) with Hemin.

3. Don't forget about microaerophilic condition (5% O2, 10% CO2, 85% N2)

4. Be careful about the thickness of your agar medium as it determines the water content of the medium.

Good luck!

Hi,

I agree with Dr. Salaheen

according to himedia, the campylobacter colonies should be grey in color and should appear spiral on gram staining under microscope. I am getting white or transparent colonies and arenot spiral..if anyone has their own microscopic image of campylobacter please share..

Amphotericin B and vancomycin (final conc. 10 μg/ml for both) should be added in your medium to make sure the bug you isolate is Campylobacter jejuni.

Dear Vandana,

Following Dr. Salaheen's suggestions, since you are working with environmental samples which may have low numbers of campylobacters and high level of background flora, I would suggest considering Preston broth as an alternative enrichment medium instead of Bolton broth.

Detection using Preston broth is useful for samples that contain significant numbers of microflora resistant to third generation β-lactams, like cefoperazone which is used in Bolton broth as well as in the commonly used mCCD selective agar. Such microflora includes e.g. strains of E. coli which appear white on mCCD agar.

During isolation on selective media, in order to avoid confusion, you can also try using chromogenic media, like Brilliant Campy Count Agar (BCCA, Oxoid) and Campylobacter Selective Agar (CASA, AES) which are highly selective media and make Campylobacter reading easy, through the enumeration of brick red colonies.

Hope this will help you!

Hey, thankyou for your reply. I am using wesley broth instead of bolton.