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Nico
01/16/15 06:18 PM  
English Brett
Historically, English strong ales aged a very long time to develop a particular and highly sought after "English character." We now know that character came from Brettanomyces, and possibly multiple strains of it. I want to brew these historical beers, but I need these English Brett strains. The only commercially available one is White Labs Brett C, which I will use, but I want more. I've asked six yeast labs and nobody has them. I'm planning on culturing it from Gale's Prize Old Ale, from Le Coq Imperial Extra Double Stout, and from Perennial's Stefan (Brett Anomalus). That's 4 English Brett strains all together, but it would be great to get my hands on any more that I can. My questions are: 1. Has anyone isolated some English Brett strains I can use? 2. Does anyone have access to these bottles and would be willing to send some my way?: Colne Spring Ale Pre-reboot bottles of Courage Imperial Stout Thanks very much! Nico
Ingo
01/20/15 01:53 PM  
Re: English Brett
https://catalogue.ncyc.co.uk/
http://www.dsmz.de/home.html
http://www.cbs.knaw.nl/
http://www.hefebank-weihenstephan.de/ (they have more than they publish, ask Ulrich, he's the chef)
SteveG
01/23/15 11:45 AM  
Re: English Brett
Wow man, quite the list of resources!!
TheGarv
03/15/15 10:20 AM  
Re: English Brett
Samuel Smiths Yorkshire Stingo is fermented with brett and is bottle conditioned. I'm culturing dregs at the moment and, as far as I can tell, my starter contains brett.

Also, the Harvey's imperial stout doesn't contain brettanomyces, I don't think. The wild strain in their culture is, allegedly, a strain of debaryomyces (crazy extremophile yeast that turns up in sausages and cheese, mostly). I cultured some and sent it to a friend from ratebeer who's doing a science on it at the moment. Tests so far are consistent with debaryomyces, but hopefully he'll have something more conclusive soon. It's a nice strain, whatever it is.
awfulperson
03/22/15 07:49 PM  
Re: English Brett
Hey Nico,

Try and seek out the beers "Tsar Bomba" and "Grain and Grape" by Buxton Brewery. These both contain Brett cultured from old bottles of Courage RIS.

TheGarv - I might have to have a go at culturing Stingo dregs! Oh, and please do post the results regarding the Harveys.
TheGarv
03/24/15 08:46 AM  
Re: English Brett
We've got some preliminary results that he posted on ratebeer, which I'm sure he won't mind me reproducing here:

"Iíve got stuff up on plates and in tubes.

What I did-

I made 1:10 serial dilutions of what Gareth sent me in a medium made of Yeast Extract, Malt Extract, Peptone, and Glucose (YPGM). From the 10^-3 dilution, I streaked onto agar plates made of the same medium+1.5g/L agar powder. Tubes were incubated at 25, and plates were incubated at 25, 30, and 37 degrees C.

All of the tubes show growth, with the 10^-1, 10^-2, and 10^-3 showing pelicle formation. There is only slight turbidity in the 10^-4 tube, so Iíd expect once the cell number get high enough we will also see a pelicle there.

I also got two distinct colony morphologies on the plates incubated at 30 degrees. Only one type of colony was present on the 25 degree plates, and no growth was observed on the 37 degree plates. The colonies on the 25 degree plates are identical to one of the morphologies on the 30 degree plates, so we are dealing with a mixed culture of at least two organisms. For now, Iím calling them "Deb 1" and "Deb 2". Deb 2 looks like run of the mill Sacch to me, whereas Deb 1 looks like some of the environmental isolates Iíve cultured before; Flat, spreading, white-grey in color.

Because Debaromyces is supposed to be very resistant to chlorine dioxide, I used both isolates, the sample Gareth sent me, and one of the dilution tubes to set up a series of incubations with or without chlorine dioxide at an initial concentration of 0.3mg/L. If one of the isolates grows, that would be great. Otherwise, the chlorine dioxide will serve to enrich the mixed population for whatever (very) small number of Debaromyces are there, and then I will attempt again to isolate on a plate."

Then an update:

"Little update.

I ran PCR on the two isolates I have using a technique nearly identical to the one described in this blog post by another person keen on beer microbiology: http://beer.suregork.com/?p=3598

Turns out this technique has been used a number of times for the rapid identification from brewery "contaminants"- See Evaluation of ITS PCR and RFLP for Differentiation and Identification of Brewing Yeast and Brewery ĎWildí Yeast Contaminants and Sequence-based identification of species belonging to the genus Debaryomyces for examples.

Here are the results of that PCR (Iíve inverted the colors, as I prefer to see the bands in black...plus, it saves on ink if/when I print the picture for my notes!)

[Here's a link to the picture - not sure how to embed it here: http://i.imgur.com/kjqdgPu.pnggarbagehttp://i.imgur.com/kjqdgPu.png]

Lanes 1 and 6 are 1KB ladder from NEB.
Lane 2 is a Brett isolate from WLP.
Lane 3 and 4 are the two isolates from the sample I was sent.
Lane 5 is an isolate from Black Label Gueuze that I wanted to identify (A Brett, for reasons that you should be able to work out by the end of this)

Lane 2 shows me that the primers and my PCR protocol work (on Brett at least).

Lane 3 and 4 indicate that the two isolates are distinct (pretty easy to tell from morphology too), and that neither of the isolates are likely Brett, due to the different size bands.

Ideally, I would have had a Sacch isolate to run as well, but I didnít have any grown up, but I think that itís a fairly safe bet that Lane 4 is a Sacch.

Lane 3 Iím not so sure on. BUT, suffice it to say that itís NOT the size that Debaromyces has been reported to be using these primers.

So, Iím currently working on doing some other stuff to try to pull out what Debaromyces might be present in the samples.

Stay tuned!"

And:

"Oh, I should mention that I didnít do the restriction enzyme digestion due to not having the specific enzyme needed. So only the PCR was done."

Apparently he's currently enriching on xylose to try and pull it out.
awfulperson
04/06/15 09:28 PM  
Re: English Brett
Thank you for posting! I found the thread... please post an update either here or on RB if you do a single fermentation with it!
awfulperson
04/06/15 09:40 PM  
Re: English Brett
Oh, another thought

Nico, if you can, try finding Empire Pale Ale from Burton Bridge Brewery. Unfortunately I can never find anything from them in my area (Manchester, UK) so I don't get to drink it frequently, but the taste definitely implies the presence of some non-Saccharomyces stuff. TheGarv, you too should try it if you can find it locally!
 
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