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Simple Yeast Storage Procedure

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@WoodlandBrew,





Shoud I follow the calculator or the volume of slurry?


I know precisely what you mean. Lately I have been cold crashing and decanting my starters into a few mason jars for multiple uses and my measured volume usually falls way short of what my calculated amount should be.

In some cases I would have to assume 5 billion cells per ml to get me in the ballpark. I know I lose a little bit when decanting but not enough to make up the difference. I've started relying more on volume and keeping notes of yeast strains, starting volume, age, starter size and that sort of thing. It's a tough business.
 
That's right. I have seen a significant difference when I failed to allow the temperature to stabilize (about double)
A couple years back I read an article that said that cold pitching was not a problem. That a more serious issue could come from yeast being warmer than the wort it's pitched into. (Say, a summer day where air temperature could be +85° going into 60° wort.) Since then, I've been pulling my yeast from the fridge after starting to transfer wort to my fermenters. I decant all the liquid from the jar and immediately top it off with fresh wort coming from the kettle. I leave that sitting, unmixed until all the rest of the wort has been transferred and oxygenated (10-15 minutes). Then I stir it up and pitch.

For the couple of years that I’ve been doing this it’s worked well. Your quote above is making me rethink my process and that I need to do a side-by-side.
 
I've measured fermentation performance on cold pitched yeast vs equal temperatures and the cold pitch took twice as long to start, but growth rate and biomass yield were about the same. How this effects beer quality I didn't try to quantify.
 
I would suspect that it may be yeast dependent. My house yeast (Pacman) works well at low temperatures which could lessen the impact.

The first generation (edit:and second) of a Saison (Wy3726) was a little slow to start, but had no problem with attenuation or flavor using the cold pitch method. But a late night pitch of the third generation of that same yeast had full krausen by morning. In fairness, I probably over pitched and moved 65° wort temperature carboys into an 80° water bath. I'm guessing that made for a smooth transition to fermentation temperature.
 
Ok so I dont need to go through the whole boil and cool water then mix with slurry, let separate into layers then decant into my ball pint jars. You mean I can just fill the jars with slurry and let it separate while it cold crashes, decant liquid, then make starter. I'd love simpler method than what I've been doing which I think is the traditional way of washing yeast. I have a batch off 001 I've been reusing since dec mostly on 1.062 and lower beers and I'd love to keep more of it around. So am I looking at this right.
 
How many generations of slurry harvesting is too much?
I've gone 6 generations of repitching slurry with no problems but I wonder if it would be safe to go more?
 
What are the negative effects of keeping the mason jar lid on too tight?

Off gassing Will produce some pressure. I usually tighten it down (in 1/2 pint jars) then loosen it once a night to let out the pressure. Do that every night or so for a week and it is all good. Note: don't open it all the way, Just unscrew the band and that ought to do it.
 
Adding my unscientific observations.

I've been harvesting yeast for many years. My technique is not that much different than yours. The difference is I transfer all the beer possible out of my carboys. By tilting and careful placement of the racking cane there's only maybe and ounce of liquid left. I then add boiled and chilled water, swirl, pour into mason jars and store refrigerated. No additional rinsing.

edit to add:
I generally use 1.5B cell/ml as a starting point when estimating pitch rates.

I feel a certain amount of joy that I came on today with a question and rather than post an entirely new thread, I obsessed that I would find the answer in another thread, and I succeeded.

I have been washing yeast using the sterile water / mason jar technique for a little over a year and the harvest on 4/4/15 from a recent brew netted me:

1 quart mason jar, roughly half full of yeast

2 pint jars with around 150 - 200 mL (if the graduated markings on the side are to be trusted) of yeast in each.

So.... when it comes to using this yeast on May 2nd (AHA BIG BREW DAY), I have a couple questions.

1) Is the bold comment from above a safe starting guess as to the slurry percentage?
Brewers Friend Yeast Calculator allows a range of 1 to 10 billions cells per mL
Mr. Malty has a sliding scale and the default setting is 2.4 B/mL

What is a safe starting guess for the percentage of my slurry?

2) I am using this yeast roughly 1 month after harvesting, so I will assume the "industry standard" of 20% viablility loss per month?

026.jpg
 
I do BIAB and use pellet hops, and do nothing to remove cold break material when I pour into the fermenter. The result is that I have a lot of trub, and no way to remove it, so I've avoided yeast washing, etc. On the other hand I have all wide mouth fermenter, and I top crop nearly every brew several times using a sterile ladle. I've had excellent results with top cropping.... There are huge amounts of yeast in the krausen, and it's extremely viable and I suspect young yeast.
I use rubbermaid screw top pint containers rather than mason jars, as the lids are not subject to rust......... It's worked extremely well for me and takes little time.

H.W.
 
How many mls of liquid do you extract when you top crop? What do you think the cell count of the top crop is? I have used washed and unwashed in a wide mouth fermentor but haven't bothered to open and top crop
 
I've never measured ML of liquid........ I try to avoid the liquid as much as possible and just skim the krausen. The settled yeast cake I would estimate at about 80 ML from a 2.5 gallon brew..... I normally crop several times. to get this. I've pitched 1/4 of this amount and had active fermentation within an hour, so it's a LOT of very active young yeast cells in my opinion. I've never done yeast calculations, and I don't think they would be accurate in comparison to bottom harvesting as there is zero trub mixed in. The liquid has almost no alcohol in it due to the fact that I take my crops at around 24 - 48 hours from pitch..... it's pretty much just unfermented wort.


H.W.
 
How many generations of slurry harvesting is too much?
I've gone 6 generations of repitching slurry with no problems but I wonder if it would be safe to go more?

It never gets un safe. I have a batch of US-05 that is on its 26th generation. The only "problem" is that the yeast will mutate and change over the generations. I like this because it is no longer US-05, it is Cottonwood Breweries house blend ale yeast.

If you want to keep the natural yeast flavor and performance, I would recommend no more than 5 generations before it starts to change its profile
 
It never gets un safe. I have a batch of US-05 that is on its 26th generation. The only "problem" is that the yeast will mutate and change over the generations. I like this because it is no longer US-05, it is Cottonwood Breweries house blend ale yeast.



If you want to keep the natural yeast flavor and performance, I would recommend no more than 5 generations before it starts to change its profile


Have you noticed any difference in your house yeast vs. the original strain?
 
It never gets un safe. I have a batch of US-05 that is on its 26th generation. The only "problem" is that the yeast will mutate and change over the generations. I like this because it is no longer US-05, it is Cottonwood Breweries house blend ale yeast.

If you want to keep the natural yeast flavor and performance, I would recommend no more than 5 generations before it starts to change its profile

Wow, 26 generations? What kind of mutations or differences are you seeing?

I'm on my 5th generation with WLP 835 Lager X and I'm thinking of going 5 more since the yeast is only available once per year.
 
What is a safe starting guess for the percentage of my slurry? I am using this yeast roughly 1 month after harvesting, so I will assume the "industry standard" of 20% viablility loss per month?
I've been harvesting yeast for a long time and have come to a lot of the same conclusions through trial and error that WoodlandBrew has come to using actual science. Bought his book on Amazon a couple of weeks ago and have been trying wrap my machinist brain around it. A couple of general takeaways. "... for beer a 50% error pitch rate will likely go unnoticed." "Viability does not drop much over time, do don't use any date-based calculations for viability."

The cell count in a slurry can vary a huge amount. I think the 1.5B cell/ml that I use is conservative. Especially for a relativity clean slurry, but I'd like to hear Steven's opinion on that and what number might be better.

Nice pictures! Looks like you put about the same care into keeping debris out of your fermenter as I do.
 
Have you noticed any difference in your house yeast vs. the original strain?

not much, mabe a little bit. I guess I should split a batch and do a side by side comparison to see the real differences. The main reason was to save $5 a brew, not buying yeast
 
I've been harvesting yeast for a long time and have come to a lot of the same conclusions through trial and error that WoodlandBrew has come to using actual science. Bought his book on Amazon a couple of weeks ago and have been trying wrap my machinist brain around it. A couple of general takeaways. "... for beer a 50% error pitch rate will likely go unnoticed." "Viability does not drop much over time, do don't use any date-based calculations for viability."

The cell count in a slurry can vary a huge amount. I think the 1.5B cell/ml that I use is conservative. Especially for a relativity clean slurry, but I'd like to hear Steven's opinion on that and what number might be better.

Nice pictures! Looks like you put about the same care into keeping debris out of your fermenter as I do.

If by Steven, you mean Woodland Brew, then this is his take on this question:
A safe guess is 1 billion per ml. The actual content depends on how much trub is in mixed in. If it is all grain with everything poured from the brew kettle to the fermentor it will be close to 1 billion per ml. If it is extract, or care is taken to prevent protein from being transferred to the fermentor then it is closer to 2 billion per ml. The viability will be dependent on the amount of alcohol in the beer that it was harvested from. (search my blog for "abv effect"

I was worried that my question would get lost in this thread, so I decided to ask the source.
 
I was worried that my question would get lost in this thread, so I decided to ask the source.
Nice! Always good to hear from the source. From the look of your slurry, it doesn't appear that you have much trub. Looks similar to what I harvest. So, I guess the 1.5B cells/ml number is pretty close for what we have. Did you send a picture to WoodlandBrew? I'd like to hear his take after seeing your harvest.
 
I didn't send the pic to him, but maybe he will see it in the thread?

I played it conservative and went with 1.2B /mL
 
Sorry I haven't been in this thread much recently. (Sometimes life gets busy) The amount of hop debris can give you an idea of the cell density, but it's often difficult to estimate the protein trub from a picture.
 
So if I wanted, I could just pour the yeast cake from a previous batch (trub and all) into a sanitized mason jar and pitch that yeast, totally unwashed, on my next beer? This sounds too good to be true.

And no starter may not be needed depending on the abv of the last beer that cake was used on?

What about just make an over-sized starter, and pouring the excess in a sanitized mason jar? Could I do that to avoid trub and/or washing? If I have to make a starter with the unwashed yeast, then why not just do an over-sized starter from the get go?


Going back to the multiple generation part (mainly Terek's "house" yeast). If you use a certain strain enough to where it does mutate (and the new strain is favorable over the old), how do you keep the current mutation from going through a second mutating phase and thus change that yeast's profile yet again? If a certain profile is not available, how do you maintain a yeast's current profile so it doesn't go through some sort of mutation?

What a great thread!
 
So if I wanted, I could just pour the yeast cake from a previous batch (trub and all) into a sanitized mason jar and pitch that yeast, totally unwashed, on my next beer? This sounds too good to be true.

And no starter may not be needed depending on the abv of the last beer that cake was used on?

What about just make an over-sized starter, and pouring the excess in a sanitized mason jar? Could I do that to avoid trub and/or washing? If I have to make a starter with the unwashed yeast, then why not just do an over-sized starter from the get go?


Going back to the multiple generation part (mainly Terek's "house" yeast). If you use a certain strain enough to where it does mutate (and the new strain is favorable over the old), how do you keep the current mutation from going through a second mutating phase and thus change that yeast's profile yet again? If a certain profile is not available, how do you maintain a yeast's current profile so it doesn't go through some sort of mutation?

What a great thread!

1) Yes. My understanding is that you wouldn't want to use the yeast from a darker / hoppier beer to make a lighter / less hoppy beer as it may taint the color / flavor.

2) according to the blog, indeed.

3) yes. Each and every time that I buy a smack pack or vial of liquid yeast, I do exactly that.
Make a 2L starter, crash and decant and split it in half. Make a 2L starter for beer and preserve the other for future use.

4) I will leave this answer to Woodland.

5) I agree.
 
I'm using 3 gallon Walmart ice tea dispensers mostly....... A wonderfully designed container for brewing as far as I'm concerned........at least for my 2.5 gallon brews. The top is larger than the bottom and the lid seals with a taper that works very well.

Normally I top crop........... but this weekend I plan to "bottom crop" a lager using a stainless steel ladle after racking the lager to one of my cold crash containers that will be used to lager it.

With this lager......... and I presume most if not all lagers, the yeast layer is distinct on top of the trub layer. I typically have quite a bit of trub due to using a fairly fine crush and not making any effort to remove cold break or hop material. I use pellet hops.

My idea is to simply use the ladle to skim the yeast layer off the trub layer. There are clearly multiple layers, and all have yeast, but the bottom ones appear to have the most course trub, and the upper layers the best looking cleanest yeast.

Does this make sense to anybody?


H.W.
 
Going back to the multiple generation part (mainly Terek's "house" yeast). If you use a certain strain enough to where it does mutate (and the new strain is favorable over the old), how do you keep the current mutation from going through a second mutating phase and thus change that yeast's profile yet again? If a certain profile is not available, how do you maintain a yeast's current profile so it doesn't go through some sort of mutation?

There are a few different ways that yeast will change over time.

You may have heard that "yeast doesn't reach its full stride until the second or third batch." The gravity of the wort, malt content, acidity, mineral content of the water, and other factors affect the metabolism of the yeast. In chemostat cultures this generally takes five volume changes, or doublings of the yeast. In batch cultures (like fermentation of beer) it can take a few batches. The genome itself doesn’t change, but the gene expression does. This can be observed in the mRNA and protein content of the yeast cells.

(When brewers refer to the performance of their house strain as different from the original, this is often what they are referring to)

Another form of adaption is competition. If there are multiple strains of yeast (and possibly bacteria) then, over the course of time, the balance between these will reach some stasis, often with one completely dominating the other. This can take about a dozen batches of beer to stabilize.

The next most common form of adaption in yeast is modification of the genome by mating which requires ‘a’ and alpha haploid cells. Most brewing yeast is diploid and reproduces only by meiosis which makes the genome very stable. It’s actually rather difficult to create hybrid brewing yeast through mating.

The least common form of adaption is genetic mutation. When this does occur it is normally from a transcription error, but it can also be forced using a variety of methods (See Brewing Microbiology) When these mutation do occur they often have no impact on yeast performance, or change in a way that is not advantageous and therefor cannot compete with the original strain. The rate at which these mutation occur are 0.27% per cell division. [1] In one batch of beer the yeast doubles (divides) about 4 times. Given these rates it would take 2 million batches of beer for a yeast to mutate from an ale yeast to sake yeast. [2]


(A while back I read an article written by a laboratory manager at a very large brewery where they used gel electrophoresis to “finger print” the yeast that they were using and found that there was no genetic change over a 5 year period.)

Most often yeast is used for 5-7 batches of beer and then a new pitch is grown up from a frozen sample. This is more often done to combat contamination, rather than genetic mutation. In any brewery, contamination is inevitable.

The stock culture is typically cryogenically frozen in an aqueous glycerin solution. When these stock cultures run out new cultures are grown on pettri dishes and singe cell colonies are selected, tested, and then propagated to make a new stock.

[1] Drake, John W., et al. "Rates of spontaneous mutation." Genetics 148.4 (1998): 1667-1686.
[2] Fay, Justin C., and Joseph A. Benavides. "Evidence for domesticated and wild populations of Saccharomyces cerevisiae." PLoS genetics 1.1 (2005): e5.
 
What about these?

Ziplock.PNG



Ziploc Twist N Lock



They look great to me........ The ones I use come from Walmart, and actually are threaded. One reason I like plastic is the low thermal mass compared to glass. I can throw one of these in boiling water to sterilize it, and by the time I have the ladle sterilized and the lid off the fermenter, it's cool enough to ladle into.

H.W.
 
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