I've got isolated strains of C. jejuni and I want to culture them, If anyone have prior experience with Campylobacter jejuni, please suggest me which media will be best to use. As there are many growth media's available, I want to know the best one which is used for already isolated C.jejuni samples not those selective media's which are required for C. jejuni isolation from fecal samples, contaminated water etc.
hello Sharma,
I haven't worked with C. jejuni before. However based on papers I have reviewed , I think Mueller Hinton broth and agar support the best C. jejuni growth.
http://www.ncbi.nlm.nih.gov/pubmed/18729058
You could also consider making conditions of growth a bit Microaerophilic because this allows for more luxurious growth.
I hope I have been able to help a bit.
Cheers.
A selective blood agar medium (Skirrow's medium) can be used and growth under microaerophilic conditions at 42 degree Celsius.
Sharma
If you have isolated strains you don't need selective medium. You can use it but a simple columbia blood agar will work just fine. The most important condition when working with C. jejuni is the microaerophilic atmosfere. If you can assure a good microaerophilic atmosfere you wont have any trouble growing it. You can also grow it at 35ºC-37ºC.
C. jejuni grows at 42ºC but this temperature is used as a selective tool to stop or at least reduce the growth of most Enterobactereaceae in feacal samples so you can also grow it at 35ºC-37ºC.
I wouldn't open the microaerophilic jar in the first 48h because some strains are very susceptible to O2. After the 48h period you can check the plates for growth and reincubate them if necessary.
Sometimes we can get some contaminants in the culture plates but if you need to recover the C. jejuni strain from those plates I would advise you to use CCDA medium (Charcoal Cefoperazone Deoxycholate Agar) or the modified version of it. It's a great medium to recover contaminated strains.
Hope this helps.
Thank you Joseph, Rushikesh and Joao for your suggestion, I'll see which one medium should I use for my experiment.
I believe blood agar plates at 37 in 7% CO2 incubator will work well if you lack easy access to specialized growth chambers.
The best way is that Mueller Hinton (MH) agar with CSS (Campylobacter Specific Supplement) at 42 c in microaerophilic condition.And columbia blood agar is also good. For saving money ,you also can use blood agar.
good morning Sharma,
no direct experience, I know FSA (Food Standard Agency ) UK refers to ISO norm 10272-2:2006 Microbiology of food and animal feeding stuffs — Horizontal method for detection and enumeration of Campylobacter spp. — Part 2: Colony-count technique.
KR,
valeria
Thanks to everyone for your valuable suggestions, I'll look forward to purchase the best one medium among your different suggestions.
Sharma, I used Mueller Hinton Agar /Mueller Hinton Broth(Oxoid) supplemented with trimethoprim and vancomycin as a selective media for C.jejuni at 42 oC in microaerobic (10% O2 and 7 % CO2 and 83% N2) condition for 3-5 days.
Regards, Dr Abdullahi
Thanks to everyone, Thanks Dr Abdullahi I also think Mueller Hinton media would be best for culturing C.jejuni
You can use a selective media BBLTM Campylobacter CSM Agar (Charcoal-Based Selective Medium) for the Campylobacter jejuni.
mCCDagar (with supplement) is best media for campylobacter isolation.
Mueller Hinton (MH) agar with CSS (Campylobacter Specific Supplement) is best.
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