Cantillon Iris (2007) Wild Yeast Isolation

Some time ago I got my hands on a bottle of 2007 Cantillon Iris. While the beer, in my opinion, is one of the worst ones I’ve had (dry-hopped woody vinegar hehe) the main purpose for getting it was the analysis of the dregs because it would be totally awesome to brew something with the wild yeast floating around at that brewery.

The procedure was pretty simple: Collect the dregs, shake it gently to homogenize, dilute 1:100 and plate. After waiting 3-4 weeks for something to grow, nothing did and I threw the plates out and almost gave up. As a “just in case” attempt, I decided to plate straight dregs and voila! I got colonies. For this purpose milk lactose + bromcresol green as well as brilliant green agar plates were used (sorry no pictures) resulting in three colony types: matte, rough colonies (C1), matte round colonies (C2), and smooth, round, glossy colonies (C3). I am still not sure whether the first two are the same or not as I suspect one may just be the later stage of the other one, but perhaps they are not due to some observations. The colonies were picked and used to inoculate 5mL of LB. The rough colonies formed pellicles in the tubes while the smooth one did not. Pellicle resulting from rough matte colonies fell away after a couple days, while the round matte one persists to this day. Subsequent plating didn’t give me much for some reason and I will have to replate. Meanwhile, here are some pictures from the liquid cultures.

It seems to me that cells grown in suspension culture have a different morphology than the ones grown in plates. Everyone who’s done cell culturing knows this, but I didn’t think it was true for yeast too. From what I see, though I did not show pictures, cells from plates tend to be shorter and rounder than the ones from tubes. If anyone has any ideas or knowledge, please share.

I also noticed that the rough colonies flocculate well and the medium is crystal clear with a sediment and a pellicle, while the smooth ones, while also forming sediment, remain suspended resulting in turbid medium. That is interesting because I noticed similar behavior and colony morphology from WY Berliner Weisse Blend Brettanomyces strain, but the cells look different.

They also smell different. C1 smells very slightly funky, C2 is quite funky, and C3 just smells like medium. Interesting…

Advertisements

12 thoughts on “Cantillon Iris (2007) Wild Yeast Isolation

  1. I did the same experiment a year ago. I used Gueuze Fond Tradition from St Louis, and also noticed the same regarding plating, I had to plate the dregs directly.
    A lot of the colonies were grey and had hyphae. Regarding cell morphology, I have to say I would not know. I noticed though, that when I switched from shaking cultures (aerobic) to fermentative growth (anaerobic) the cells that grew in hyphae (black clumps) turned to “normal” yeast growth.
    I isolated 4 yeasts from this, one was Brett, the others were not really classified.

  2. Nice post!

    You might want to try something other than LB. Luria Broth is very rich and specifically made for growing bacteria, not wild yeast. I have had good success with MYPG and you may have different type of cell growth.

    J

    • Hey! Thanks for the suggestion. I used LB for liquid culture because… well because that’s what I have in hand right now. I just prepared 100 ml of LB in the lab and split it into 5 ml tubes because it was getting kind of annoying to do this with malt extract at home. I will try using a different liquid medium and see what I get. Thanks again for the suggestion.

  3. Pingback: Free Brettanomyces! « BKYeast

  4. Pingback: Free Brettanomyces Part 2 « BKYeast

  5. I want to apologize for my delay in feedback. C1 is a winner. I found out this morning that the Saison I brewed with C1 took first place in the Bruery Batch 1000 contest Belgian & French Ale category and was runner-up for Best of Show. (www.thebruery.com/batch1000) Thanks for sharing! I’d love to send you a bottle if you can send me your address. jonathan dot moxey at gmail dot com

  6. I have recently pitched a starter of c2 into a wort of around 1.050 OG. This is not a dextrin heavy wort, mostly 2 row, I built a 2L starter for it and was wondering what kind of time frame I may be looking at for a primary fermentation? The starter tried to form a pellicle each time I stopped stirring to step up so I know it’s growing, just curious.

    • I’d say around a month, though other people told me they had a complete fermentation in as little as 2 weeks. In my experience, they keep going for around 3 months until they reach complete terminal gravity.

    • It varies. Vials are 50mL and cell counts vary from 5 to 70 billion cells. When I send out vials for late comers, they’re usually 15mL with 0.5-5 billion cells.

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s