It is an anaerobic bacteria and I am finding it difficult to isolate it, we have collected skin samples from people suffering from acne.
Hi,
I don't know if you have managed to solve the Propi isolation problem yet, but I find Thioglycollate Broth an excellent media for enhancing the growth of Propi (and other skin flora) from swab or tissue cultures. After 5-7 days (although it may take up to 9) at 35-37C, if growth is detected in the Broth, subculturing to 5% sheep's blood agar is always helpful to me to categorize and isolate Propi. I usually incubate two plates, one aerobically and the other anaerobically just to help me to categorize quickly. Hope this helps!
If your bacterium is anaerobe, you can use anaerobic or microaerophilic conditions for growth. Enriched medium would give better results and for further identification you may perform molecular methods
Another useful enrichment mediumf for isolation of Propi is Fastidious anaerobic broth
It is difficult to harvest and grow the anaerobic bacteria. Anaerogas Pack/anaerobic conditions is mandatory while harvesting the microbe. Brain Heart Infusion (BHI) media containing 1% glucose is ideal for the growth in anaerobic conditions. You can refer: "Nano-colloidal Carriers of Isotretinoin: Antimicrobial Activity against Propionibacterium acnes and Dermatokinetic Modeling.Mol Pharm. 2013"
inoculation in to routine media like blood agar and incubation in anaerobic jar (Dynamicro) will be sufficient for isolation
It is should be sub-cultured every day and should be cultured in anaerobic gas pack (Anaerogas pack 3.5L, M/s Hi Media Lab. Ltd., Mumbai, India) at 37˚C for 48 h.
It should be sub-cultured every 7th day and should be cultured in anaerobic gas pack (Anaerogas pack 3.5L, M/s Hi Media Lab. Ltd., Mumbai, India) at 37˚C for 48 h.
Skin swabs work well for surface Prop acnes, but tell you nothing about follicular P. acnes. For the latter follicular casts have been used. Check out:
Sampling and detection of skin Propionibacterium acnes: current status.
Alexeyev OA, Jahns AC.
Anaerobe. 2012 Oct;18(5):479-83. doi: 10.1016/j.anaerobe.2012.07.001
It depends on the context of the sample and if it is a clinical specimen you want to isolate from, or an already pure isolate. If you have an isolate you want to grow up then anaerobic blood agar works very well.
Can we use human blood as supplement instead of sheep blood for the culturing of Propionic bacteria?
If yes, kindly refer the article
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