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Isolated Yeast (Tree House): How to Identify and Characterize?

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Lakeside_Brewer5

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I havent had many TH but the ones I did really have a hefe/wheat beer smell to it.
I poured a bit of a belgium wit bier in my neipa and wasnt impressed, the lower abv probably makes it thinner then you want so I dont know if this is worth comparison.
To me this is evidence that it’s not actually 2 different beer styles, but the same exact wort, mashed and sparged, then split into a main batch and small batch (or 1 main and 2 small batches with the right equipment) for each yeast to interact with the wort in its own way.
 

echoALEia

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@echoALEia Ive asked this before somewhere in HBT with no response though. How do you know how many cells are in a packet of SO4 to know you are under pitching? Fermentis states is 6B cells per gram, but if you go by Mr. Malty standards, its 20B cells. So this is tremendous variation. What do you go by to know you are under pitching and what do you think your pitch rate actually is? Thanks.
Hey sorry! Didn’t see the post from before. I guess in the end I really don’t. I just go by recommended pitching rates in beersmith then pitch less than that. For this batch 137g was recommended. I pitched 90g. No rhyme or reason why. Again, sorry about no responding before
 

echoALEia

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LOL. If you don’t want to share your recipe bud, don’t share your recipe. There’s been a lot of people who contributed to this thread, who shared beer, who shipped beer to be test, etc. You through out an idea that was also thrown out about 80 pages ago. Nothing wrong with that, but let’s not act like you’re the only one contributing.
This seems more process oriented than recipe oriented, in my opinion. He has laid out good/detailed ideas for a process.
 

troxerX

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To me this is evidence that it’s not actually 2 different beer styles, but the same exact wort, mashed and sparged, then split into a main batch and small batch (or 1 main and 2 small batches with the right equipment) for each yeast to interact with the wort in its own way.
Gosh, wanted to ask, you read through the entire thread?, LOL I stopped by panel #13 and re-started at #90...

Affirmative, all options are on the table, either a wort that we split and ferment separately and then re-inject or multiple worts fermented separately and then blended. One of the reasons I lean on different worts is that it gives you the capability to maximize its properties for the yeast you will be pitching and also allows you to add layers of texture and complexity vs one wort with a fixed set of properties. It also allows you to add more layers of different hops without overhopping/overbittering, dilution may play a role in saturation but we have to figure that one out. What I can’t think of right now is that if aroma is impacted. If splitting a wort, it has to be one properly designed to add all the sensory just like @echoALEia did. TH has multiple beers and who knows all these options are part of their processes. If let’s say some of their beers are made with multiple worts using the same principles, I might think they can use that to their advantage by brewing a huge large base batch and then split into different blends with their own other smaller batches, people see blending as a limitation, whatever approach we take I see it as an opportunity.
 

Clyde McCoy

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Summary of findings:

The primary yeast in Julius, jjjuliusss, and Very Green matches the electrophoretic banding profile of Fermentis S-04 (as identified by @isomerization):

Screen Shot 2020-09-20 at 4.09.15 PM.png


Screen Shot 2020-09-20 at 4.09.28 PM.png


Screen Shot 2020-09-20 at 4.09.43 PM.png


Screen Shot 2020-09-20 at 4.10.02 PM.png


In fact, S-04 was the ONLY strain I could find until @Northern_Brewer suggested I try WLN agar. This is what various Tree House beers look like on WLN:


Screen Shot 2020-09-20 at 4.11.57 PM.png
Screen Shot 2020-09-20 at 4.12.15 PM.png

Screen Shot 2020-09-20 at 4.12.33 PM.png
Screen Shot 2020-09-20 at 4.12.52 PM.png



Everything on these plates tends to match the electrophoretic banding profile of Fermentis S-04, except for the large, beige colonies, which are 3-4 different strains:


Screen Shot 2020-09-20 at 4.15.57 PM.png


Screen Shot 2020-09-20 at 4.16.05 PM.png


(continued on next post, max 10 images)
 

Clyde McCoy

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Screen Shot 2020-09-20 at 4.16.12 PM.png


Screen Shot 2020-09-20 at 4.16.36 PM.png


Screen Shot 2020-09-20 at 4.16.52 PM.png



These strains are likely WB-06 and T-58 (as identified by @isomerization), but I’ll say these are unconfirmed because the profiles aren’t an exact match and may require a bit of PCR optimization to say more conclusively.

I’m again going to quote Chris White (White Labs) on mixed-strain fermentations:

“When yeast is pitched into beer, it starts to grow, entering into a log phase of growth after a few hours. This is when the bulk of the flavor compounds are produced. 12-36 hours into the fermentation. Therefore, if your goal is flavor, you need to add the multiple strains early on, preferably together. Note that if you just want another strain for bottling, or to complete attenuation, go ahead and add later. Very little flavor contribution happens here, unless the beer undergoes prolonged ageing”

You'll notice that on some of the WLN plates it looks like S-04 (small green colonies) is 99+% of the plate.

I'm pretty near my limit on the amount of time I can spend on this, but because the composition of yeast in the can will change over time (all beers above are typically analyzed at 1-2 months old) I am willing to test a few very fresh cans (ideally <10 days), PM me if you can mail. These will be the most indicative of the ratio of S-04 to everything else at the time of canning.
 

echoALEia

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I’m so intrigued that Monkish uses 71b. I don’t even know where to begin with the questions for that. Is that first colony S33?
 

Clyde McCoy

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I’m so intrigued that Monkish uses 71b. I don’t even know where to begin with the questions for that. Is that first colony S33?
Sorry to clarify, there is no match between Monkish and 71B (or between Tree House and 71B). It is somewhat similar to S-33, but would need to run them together. That Monkish profile might be Conan or LAIII, I haven't tested those.
 

troxerX

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Summary of findings:

The primary yeast in Julius, jjjuliusss, and Very Green matches the electrophoretic banding profile of Fermentis S-04 (as identified by @isomerization):

View attachment 699335

View attachment 699337

View attachment 699338

View attachment 699339

In fact, S-04 was the ONLY strain I could find until @Northern_Brewer suggested I try WLN agar. This is what various Tree House beers look like on WLN:


View attachment 699341View attachment 699342
View attachment 699344View attachment 699345


Everything on these plates tends to match the electrophoretic banding profile of Fermentis S-04, except for the large, beige colonies, which are 3-4 different strains:


View attachment 699347

View attachment 699348

(continued on next post, max 10 images)
Just curious, do we know why these 3-4 yeasts look beige and S04 green? is this an artifact of the test itself or is there any biological reason?
 

Noob_Brewer

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From what I understand about these agar plates is that the green is simply a dye in the agar and the yeast colonies take up the dye at different rates, so the presence of Visibly different colors of colonies points merely to the presence of multiple yeast strains.

I am unsure though if you can use thesetypes of qualitative analyses to come to a conclusion of the distribution of one type of yeast (all the small darker green colonies) vs another (larger beige colonies)
 

Clyde McCoy

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From what I understand about these agar plates is that the green is simply a dye in the agar and the yeast colonies take up the dye at different rates, so the presence of Visibly different colors of colonies points merely to the presence of multiple yeast strains.

I am unsure though if you can use thesetypes of qualitative analyses to come to a conclusion of the distribution of one type of yeast (all the small darker green colonies) vs another (larger beige colonies)
You can if you've tested lots of colonies
 

Noob_Brewer

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You can if you've tested lots of colonies
You've done a sh*t ton of work @Clyde McCoy as well as @isomerization has as well. Pretty cool watching all of this as a long time lurker in this thread. BTW....im on to you!

I'm pretty near my limit on the amount of time I can spend on this, but because the composition of yeast in the can will change over time (all beers above are typically analyzed at 1-2 months old) I am willing to test a few very fresh cans (ideally <10 days), PM me if you can mail. These will be the most indicative of the ratio of S-04 to everything else at the time of canning.
This is code for - give me FRESH beer and I'll continue with this escapade! LMFAO :) lol.
 

Noob_Brewer

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seriously though, appreciate your work on this! This isn't easy and is time intensive!
 

Clyde McCoy

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You've done a sh*t ton of work @Clyde McCoy as well as @isomerization has as well. Pretty cool watching all of this as a long time lurker in this thread. BTW....im on to you!


This is code for - give me FRESH beer and I'll continue with this escapade! LMFAO :) lol.
I use to think that until I realized how many hours I've put into this...
 

Clyde McCoy

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It escalates quickly!
I think I crossed the 100 colony mark today...

It's looking like I will identify the same minor strains as you previously found (T-58 for sure, I'm still not 100% on WB-06), but I think they might be more rare than previously thought (going by WLN here and previous testing without WLN which was a needle in a haystack problem).
 
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isomerization

isomerization

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It’s very interesting to me that the 3 other strains are metabolizing the dye. I could never find much info on WLN agar mechanics, but Chad Yackobson suggested Wit yeast were among Sacc spp that could do this, don’t know if that’s helpful or not though :)
 
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isomerization

isomerization

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I think I crossed the 100 colony mark today...

It's looking like I will identify the same minor strains as you previously found (T-58 for sure, I'm still not 100% on WB-06), but I think they might be more rare than previously thought (going by WLN here and previous testing without WLN which was a needle in a haystack problem).
That’s cool in its own right, as I could have envisioned them having to change with the process scaling up. Speaks to how difficult their process is, given the quality issues reported.
 

Clyde McCoy

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It’s very interesting to me that the 3 other strains are metabolizing the dye. I could never find much info on WLN agar mechanics, but Chad Yackobson suggested Wit yeast were among Sacc spp that could do this, don’t know if that’s helpful or not though :)
I tested dozens and dozens of small dark and medium green colonies and they were always S-04 which is why I've started to ignore them and focus on the others. Always good to include a few though to be sure.

Looking back at your gels it doesn't seem like S-04 would be confused with anything else.
 

lilbova3

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View attachment 699349

View attachment 699350

View attachment 699351


These strains are likely WB-06 and T-58 (as identified by @isomerization), but I’ll say these are unconfirmed because the profiles aren’t an exact match and may require a bit of PCR optimization to say more conclusively.

I’m again going to quote Chris White (White Labs) on mixed-strain fermentations:

“When yeast is pitched into beer, it starts to grow, entering into a log phase of growth after a few hours. This is when the bulk of the flavor compounds are produced. 12-36 hours into the fermentation. Therefore, if your goal is flavor, you need to add the multiple strains early on, preferably together. Note that if you just want another strain for bottling, or to complete attenuation, go ahead and add later. Very little flavor contribution happens here, unless the beer undergoes prolonged ageing”

You'll notice that on some of the WLN plates it looks like S-04 (small green colonies) is 99+% of the plate.

I'm pretty near my limit on the amount of time I can spend on this, but because the composition of yeast in the can will change over time (all beers above are typically analyzed at 1-2 months old) I am willing to test a few very fresh cans (ideally <10 days), PM me if you can mail. These will be the most indicative of the ratio of S-04 to everything else at the time of canning.
How long is prolonged aging I wonder? I’ve mentioned before that after some age all their beers taste the same to me. Which leads me to what White said about ‘very little flavor contribution happens here unless prolonged aging’. Haven’t had a fresh TH in awhile.
 
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isomerization

isomerization

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I tested dozens and dozens of small dark and medium green colonies and they were always S-04 which is why I've started to ignore them and focus on the others. Always good to include a few though to be sure.

Looking back at your gels it doesn't seem like S-04 would be confused with anything else.
yeah I think it’s clear that S-04 (the most abundant and closest banding match) is their primary yeast strain. I’m 75% convinced on T-58, but the WB-06 just doesn’t fit imo. Something close, or perhaps a banked strain from serial passaging of WB-06 (can you do that with a dry yeast)?
 

brewpharm Hill

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@Clyde McCoy your work is very appreciated here! Same goes for @isomerization. None of this would be possible with out lab savvy gents such as yourselves.

Considering the dominant flavors that come from T58 and Wb06 (assuming this is even being used) I feel like they're either used in seriously small percentages compared to s04 OR they're used post s04 pitching but pre 72 hr mark (Chris White states that there is pretty much no flavor contribution after that point). My grainfather is down right now due to the plug melting but once I have it replaced I will definitely be going back to experimenting with the blend as well as s04 on its own. Hoping to nail down some different temps and ratios
 

beervoid

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@Clyde McCoy your work is very appreciated here! Same goes for @isomerization. None of this would be possible with out lab savvy gents such as yourselves.

Considering the dominant flavors that come from T58 and Wb06 (assuming this is even being used) I feel like they're either used in seriously small percentages compared to s04 OR they're used post s04 pitching but pre 72 hr mark (Chris White states that there is pretty much no flavor contribution after that point). My grainfather is down right now due to the plug melting but once I have it replaced I will definitely be going back to experimenting with the blend as well as s04 on its own. Hoping to nail down some different temps and ratios
Pitch T58 a day later in order to control the esters?
 

brewpharm Hill

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Pitch T58 a day later in order to control the esters?

Yeah roughly. I'm pretty sure someone else has tried staggered pitching in this thread and they didn't really like the results so I need to go back and see how this person tried it and adjust from there. The book "Yeast" goes into a little more detail but the general idea you'll see below. My understanding is that within the first few days is when the flavor compounds are formed and that the further away from initial yeast pitch, subsequent pitches will have less flavor contribution.

An example they used in the book was that some breweries pitch their house strain which is a poor flocculator and then pitch a strong floccing yeast outside of that flavor making window to get the benefit of flocculation without flavor contribution.



"Q: Do I add the strains in the beginning or staggered?

A: Depends what you want. When yeast is pitched into beer, it starts to grow, entering into a log phase of growth after a few hours. This is when the bulk of the flavor compounds are produced. 12-36 hours into the fermentation. "
 

mcoman

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Thanks @Clyde McCoy, this is great information. Do you have the canned date vs the date these were plated? If I remember correctly when you tested the older can of very green before the WLN agag you found mostly T-58, vs this newer can has mostly S-04. Would this suggest that over time T-58 begins to take over?

I find it interesting that JJJuliusss appears to have more a higher concentration of of not S-04 yeast compared to Julius. If these were canned and plated after similar amounts of time, than perhaps this means that as hop content goes up they introduce more non S-04 yeast.
 
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isomerization

isomerization

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I would strongly recommend everyone avoid trying to draw conclusions from single cans. In order to be properly powered, @Clyde McCoy would need to evaluate multiple cans, preferably from throughout the canning process. I could be wrong, but that’s my thought process.

Question I’ve been thinking on lately, if another yeast is added after the primary strain has started fermenting (72 hr in, during dry hop, conditioning, etc), then how much replication will occur? I was under the impression that it would be very little. How does a strain then take over across time? Yeast shouldn’t be growing in a can right?
 

Clyde McCoy

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I would strongly recommend everyone avoid trying to draw conclusions from single cans. In order to be properly powered, @Clyde McCoy would need to evaluate multiple cans, preferably from throughout the canning process. I could be wrong, but that’s my thought process.

Question I’ve been thinking on lately, if another yeast is added after the primary strain has started fermenting (72 hr in, during dry hop, conditioning, etc), then how much replication will occur? I was under the impression that it would be very little. How does a strain then take over across time? Yeast shouldn’t be growing in a can right?
You can see the WLN plates above from 4 TH beers, so I think that counts as multiple? Fully agree that the most informative in terms of strain ratios will be the youngest cans.

@mcoman good questions and I will have to look at the dates on the cans and on the plates. I don't recall any big difference in can age but I could be wrong. I just don't know how the strains change over time in the can.

I think S-04 + T-58 is a good starting point.
 
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isomerization

isomerization

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You can see the WLN plates above from 4 TH beers, so I think that counts as multiple? Fully agree that the most informative in terms of strain ratios will be the youngest cans.

@mcoman good questions and I will have to look at the dates on the cans and on the plates. I don't recall any big difference in can age but I could be wrong. I just don't know how the strains change over time in the can.

I think S-04 + T-58 is a good starting point.
Sorry, to clarify, I meant from the same beer. As in we can’t make conclusions about what is in Julius verses Green.
 

Clyde McCoy

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Sorry, to clarify, I meant from the same beer. As in we can’t make conclusions about what is in Julius verses Green.
I'll test those ~12 beige colonies on the Very Green plate and see if they're all T-58 (which I suspect they are) 😂
 
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isomerization

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I'll test those ~12 beige colonies on the Very Green plate and see if they're all T-58 (which I suspect they are) 😂
Im not sure if you’re serious or jacking with me, but I meant test 4 different cans of Green, before making the conclusion that Green has a higher % of T-58 than other TH beers.
 

Noob_Brewer

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Hey sorry! Didn’t see the post from before. I guess in the end I really don’t. I just go by recommended pitching rates in beersmith then pitch less than that. For this batch 137g was recommended. I pitched 90g. No rhyme or reason why. Again, sorry about no responding before
Thanks for the info, by the way, I wasn’t actually picking on you for not responding lol I was just stating my Q didn’t get any response before. Appreciate your feedback though. It does seem though that when peeps are referring to “underpitching” without knowing really what’s in the packet, kinda makes it difficult eh? I imagine beersmith uses Fermentis values of 6B cells per gram as beersmith typically uses manufacturers recommendations.
 

brewpharm Hill

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I know some people are apprehensive about sharing their experiments and results, but that is what this whole thread is about. Want to try blending post fermentation? knock yourself out. Pitching yeast blends etc? Go ahead. But sharing results is the only way to make progress. If you want to conduct experiments and then hold your findings hostage forcing others to try your experiments then we collectively as a group will get no where. Having a summary of what has been tried would be a good place to start....
 

RTE

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Not completely on topic, but has anyone tried Equilibrium? They are distributing now and I swear the beers Fluctuation and Particle Physics both have a very Tree House-y biotransformation/ester taste. I could be delusional because I'm too lazy to make the drive to TH but these are the first beers I've tasted that somewhat have a TH house yeast taste to them. Fluctuation reminds a lot of Haze (but maybe better...). Maybe it'd be worthwhile to test of can of Eq and see if there is any interesting yeast floating in there? They ship directly PA OH DC NH VT VA and ND.
 

troxerX

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I know some people are apprehensive about sharing their experiments and results, but that is what this whole thread is about. Want to try blending post fermentation? knock yourself out. Pitching yeast blends etc? Go ahead. But sharing results is the only way to make progress. If you want to conduct experiments and then hold your findings hostage forcing others to try your experiments then we collectively as a group will get no where. Having a summary of what has been tried would be a good place to start....
LOL just teasing you guys, not holding anyone hostage LOL, I can’t share results right now as I’m still working towards them!, blending batches takes time... again - this may not be the solution after all but we have to try something different

Batch 2 with T-58

4EE2B022-E9C9-4FEA-BE4E-2F1E372EFC7E.jpeg
 

Clyde McCoy

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Not completely on topic, but has anyone tried Equilibrium? They are distributing now and I swear the beers Fluctuation and Particle Physics both have a very Tree House-y biotransformation/ester taste. I could be delusional because I'm too lazy to make the drive to TH but these are the first beers I've tasted that somewhat have a TH house yeast taste to them. Fluctuation reminds a lot of Haze (but maybe better...). Maybe it'd be worthwhile to test of can of Eq and see if there is any interesting yeast floating in there? They ship directly PA OH DC NH VT VA and ND.
I have a can of EQ "Infinite Skills Create Miracles" that I will test at some point soon
 
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