I'm trying to create frozen mid-log stocks of B. breve. Although I'm having trouble with growing B.breve in MRS broth supplemented with L-cysteine HCl. It grows really slowly, so it leads me to believe I have too much oxygen in the liquid media.
I fill the tubes up to the brim with broth, cap lids tightly and incubate in 5% CO2 humidified incubator, but still getting weird, inconsistent results.
This is bacteria know, It very slowly in growth in media , I t is need 24 hours for skim milk fermentation. But you can use some effects for growth speed e.g. growth under anaerobic condition by 10 % of CO2 and 30% N. The method very best
You still (evidently) have too much oxygen. (With the caps on tightly, it doesn't really matter what kind of atmosphere the organisms are incubated in, and the 5% CO2 isn't what you'd be shooting for, since there's still lots of O2). If you don't have access to anaerobic incubator or some type of anaerobic jar like a GasPak jar, some less expensive options are the Bio-Bag system by BD or Oxoid's Anaerogen. The only problem is those are really made to use with plates lying flat, not tubes, but in a pinch one can be creative. I've even had luck with buying the gas-generating sachets alone, and using Seal-a-Meal bags as the container. As for the medium, if the MRS + cysteine doesn't work, tryptic soy broth + 5% defibrinated sheep blood ought to do the trick.
Vance McCracken Can we grow bifidobacterium in MRS + 0.05% L-cysteine-HCl incubated in a CO2 incubator (2-20%)?
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