Homebrewers Association | AHA Forum

General Category => Yeast and Fermentation => Topic started by: TMX on February 07, 2015, 02:04:34 AM

Title: Shaken not Stirred
Post by: TMX on February 07, 2015, 02:04:34 AM
(http://i42.photobucket.com/albums/e346/TXAleWorks/Yeast/IMG_1122_zps664e3c14.jpg)

Trying the "James Bond" starter method.

Lots of messages back and forth with @S. cerevisiae to get an understanding of his process.
Here are the steps I am going by:

1.   Gather the stuff needed
2.25 L water
200g DME
Glassware
Lids
Funnel
Sanitizer
Cotton
Alcohol
Yeast
2.   Prepare and boil starter wort for 15 min; chill when complete
3.   While chilling; sanitize glass jugs, lids, funnels, and other equipment needed
4.   When chilling complete: Clean pouring lip of pan with alcohol and transfer in to 2x1g glass jug
5.   Clean yeast package / vial with alcohol and add equal amounts to each starter
6.   Cap jugs, and “Shake like they owe you money”
7.   Rest for a few min and loosen caps
8.   Pitch or crash at High Krausen

This is split to get two 1 litre starters to pitch in 10g of wort....Irish Red I am brewing on Sunday.
More to follow.

T
Title: Re: Shaken not Stirred
Post by: klickitat jim on February 07, 2015, 03:51:45 AM
I'm kinda in too. Starting my starters brew morning and pitching at HK
Title: Re: Shaken not Stirred
Post by: brewday on February 07, 2015, 04:03:16 AM
Tomorrow will be my third batch in a row using a similar method.  So far so good...
Title: Re: Shaken not Stirred
Post by: narcout on February 07, 2015, 09:01:22 AM
8.   Pitch or crash at High Krausen

I've been trying the crash at high krausen method myself (although I hit my starters with O2 rather than shaking, and I put them on a stir plate at a very low RPM).
Title: Re: Shaken not Stirred
Post by: Philbrew on February 07, 2015, 03:58:56 PM
Yeast
2.   Prepare and boil starter wort for 15 min; chill when complete
3.   While chilling; sanitize glass jugs, lids, funnels, and other equipment needed
4.   When chilling complete: Clean pouring lip of pan with alcohol and transfer in to 2x1g glass jug
5.   Clean yeast package / vial with alcohol and add equal amounts to each starter
6.   Cap jugs, and “Shake like they owe you money”
7.   Rest for a few min and loosen caps
8.   Pitch or crash at High Krausen

This is split to get two 1 litre starters to pitch in 10g of wort....Irish Red I am brewing on Sunday.
More to follow.

T
How much and what kind of yeast?

How long between steps 7 & 8?
Title: Re: Shaken not Stirred
Post by: TMX on February 07, 2015, 06:15:51 PM
Split a vial of Irish Ale yeast from a local source that rents space at the college microbiology lab in Colorado Springs decanted until there was 30ml of yeast and medium then put half in each jug 

Only have to wait a minute or so. Just to give it time for the wort to run back down so you don't get sprayed in the face when you break the seal. After that I just leave the caps sitting on top of the jug
Title: Re: Shaken not Stirred
Post by: Philbrew on February 07, 2015, 09:08:19 PM
Split a vial of Irish Ale yeast from a local source that rents space at the college microbiology lab in Colorado Springs decanted until there was 30ml of yeast and medium then put half in each jug 

Only have to wait a minute or so. Just to give it time for the wort to run back down so you don't get sprayed in the face when you break the seal. After that I just leave the caps sitting on top of the jug
How long from when you pitch the yeast in the starter to when high krausen occurs in the starter?  Do you add starter to main wort at that time?
Title: Re: Shaken not Stirred
Post by: klickitat jim on February 07, 2015, 09:50:07 PM
Split a vial of Irish Ale yeast from a local source that rents space at the college microbiology lab in Colorado Springs decanted until there was 30ml of yeast and medium then put half in each jug 

Only have to wait a minute or so. Just to give it time for the wort to run back down so you don't get sprayed in the face when you break the seal. After that I just leave the caps sitting on top of the jug
How long from when you pitch the yeast in the starter to when high krausen occurs in the starter?  Do you add starter to main wort at that time?
1. Depends
2. You bet, unless you didn't or couldn't time it right. In that case, crash it in the fridge before it ferments out.
Title: Re: Shaken not Stirred
Post by: TMX on February 07, 2015, 10:56:16 PM

Split a vial of Irish Ale yeast from a local source that rents space at the college microbiology lab in Colorado Springs decanted until there was 30ml of yeast and medium then put half in each jug 

Only have to wait a minute or so. Just to give it time for the wort to run back down so you don't get sprayed in the face when you break the seal. After that I just leave the caps sitting on top of the jug
How long from when you pitch the yeast in the starter to when high krausen occurs in the starter?  Do you add starter to main wort at that time?
1. Depends
2. You bet, unless you didn't or couldn't time it right. In that case, crash it in the fridge before it ferments out.

Mine went about 12 hours. I then crashed it and will pitch tomorrow.

I will taste the starter before I pitch.
Title: Re: Shaken not Stirred
Post by: brewday on February 07, 2015, 11:08:16 PM
The first two for me went 12-14 hrs.  Today - sometime under 12 hrs.
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 07, 2015, 11:45:50 PM
Most one to two liter starters that are made with a vial of White Labs yeast or an activator smack pack of Wyeast yeast should reach high krausen within 12 to 18 hours, sometimes sooner.  The cells are in high gear when high krausen is reached.
Title: Re: Shaken not Stirred
Post by: TMX on February 08, 2015, 02:27:47 AM
(http://i42.photobucket.com/albums/e346/TXAleWorks/Yeast/SAM_1158_zpsd0cd1ac9.jpg)

Nice little yeast bed ready to be pitched tomorrow.
Title: Re: Shaken not Stirred
Post by: Philbrew on February 08, 2015, 04:05:05 AM
Most one to two liter starters that are made with a vial of White Labs yeast or an activator smack pack of Wyeast yeast should reach high krausen within 12 to 18 hours, sometimes sooner.  The cells are in high gear when high krausen is reached.
Thanks, that answers my Q.
Title: Re: Shaken not Stirred
Post by: 69franx on February 09, 2015, 09:18:23 PM
As an example, my next brew day will be 5.5G of 1.05(targeted) Pilsner. Beersmith, MrMalty, and Yeastcalc all call for around 385B cells, give or take. If I buy 1 vial of WLP 833, split it as above into 2 1L starters, and shake the heck out of, what is my approximate yield cell mass? If I remember correctly Mark, maximum density in 1L is 200B cells, so I should yield roughly 400B, correct? Is this putting too much stress on half a vial, for a lager, asking it to go from about 50B to 200B in one step? Most of those calculators call for 2 vials and huge starters to get there: 6L un-stirred, for 1 vial, 4L for 2 vials, etc. I have 5L flask and 1G jug, so I can easily split into 2 vessels, just don't want to stress my yeast. My last 2 lagers were both under-attenuated by 5 points or so. 5 points may not be much, but they both taste sweet to me, but I know they were under-attenuated. I just really don't want a sweet pilsner. This brew day is still almost 2 weeks out, so let me know what you think
Title: Re: Shaken not Stirred
Post by: narcout on February 09, 2015, 09:50:07 PM
If you have a copy of Yeast, there is a table on page 140 which shows how many cells resulted from pitching 100 billion cells into various amounts of starter wort.

When 100 billion cells were pitched into 2 liters of 1.036 wort at 70 degrees, the result was 205 billion cells. When 100 billion cells were pitched into 4 liters of the same wort, the result was 276 billion cells.

However, the wort was not oxygenated nor was the starter agitated.  I think that would have increased the cell counts. 

I wish they had run several different iterations of the experiment to test the effects of oxygen, agitation, stir plates, etc.

Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 09, 2015, 09:58:35 PM
Is this putting too much stress on half a vial, for a lager, asking it to go from about 50B to 200B in one step?

Absolutely not, if you saw how little yeast I pitch into 600ml of wort, you would would never again question pitching half of a White Labs vial into 1L of wort. 
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 09, 2015, 10:08:22 PM
When 100 billion cells were pitched into 2 liters of 1.036 wort at 70 degrees, the result was 205 billion cells. When 100 billion cells were pitched into 4 liters of the same wort, the result was 276 billion cells.

Increasing the cell count from 100B to 205B cells requires the viable cells to double a little more than one time, which tells me that dissolved O2 was more than likely the limiting factor in this experiment.  The requirements for this method are a starter vessel that is three, preferably four or more times the volume of the starter.  Ignoring this requirement has the potential to negatively impact the results one obtains from the method.  Luckily, one gallon jugs are relatively cheap.
Title: Re: Shaken not Stirred
Post by: TMX on February 10, 2015, 02:32:26 AM
So. I pitched yesterday at about 68F around 5pm, took a quick look this morning about 7am and saw the largest krausen EVER in on of my beers.
Title: Re: Shaken not Stirred
Post by: Philbrew on February 10, 2015, 03:59:49 AM
The requirements for this method are a starter vessel that is three, preferably four or more times the volume of the starter. 
Why?  Is it the O2 available in the headspace, and cannot enriching the O2 percentage in the headspace allow one to "cheat" on the starter/vessel ratio to some degree?  I want to do 2 starters and 8L vessels are impractical and hard-to-find! (and afford!)
Title: Re: Shaken not Stirred
Post by: TMX on February 10, 2015, 05:30:33 AM
You don't need 2  8L vessels, just 2 4L vessels, or any vessel that is 3-4 times larger than the starter you want to make.

I used 2 1gallon jugs to make 2 1L starters to pitch into 10gal of wort
Title: Re: Shaken not Stirred
Post by: Philbrew on February 10, 2015, 04:16:14 PM
You don't need 2  8L vessels, just 2 4L vessels, or any vessel that is 3-4 times larger than the starter you want to make.

I used 2 1gallon jugs to make 2 1L starters to pitch into 10gal of wort
Sorry, spell checker erased an L in my post.  I wanted to say that I want to make two 2L starters (to pitch in two six gal. lager batches).  I don't have four 1 gal. jugs.
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 10, 2015, 04:55:48 PM
Sorry, spell checker erased an L in my post.  I wanted to say that I want to make two 2L starters (to pitch in two six gal. lager batches).  I don't have four 1 gal. jugs.

Headspace is critical to achieving a proper shake.  One is attempting to turn the wort into as much foam as is humanly possible.  This method is designed to be low-tech and low-cost. 

With that said, another thing that I am attempting to dispel is the insane notion that people have to hit the cell counts provided by yeast calculators in order to have a healthy fermentation.   It is better to have 60 billion healthy, ready to go to war with the wort cells than it is to have 200 billion stressed cells that are barely clinging to life.  The difference between 60 billion cells and 200 billion cells is approximately 180 minutes of propagation time.

The maximum cell density for 2 liters of wort is roughly 400 billion cells.  The maximum cell density for 5 gallons of wort is roughly 3.8 trillion cells; hence, pitching 400 billion cells requires log(3,800 / 400) / log(2) = ~3.25 replication periods.  Pitching 200 billion cells requires log(3,800 / 200) / log(2) = ~4.25 replication periods.  The notion that one has to pitch a 2 liter starter into 5 gallons of normal gravity lager wort in order to have healthy fermentation is ludicrous.  As there is more than enough carbon (sugar is carbon bound to water) in the average batch of wort to support the growth of 3.8 trillion cells, the limiting factor is dissolved O2 because the ergosterol and unsaturated fatty acids (UFA) that are synthesized by mother cells while O2 is still in solution is shared with all of the daughter cells that are created after O2 is depleted.  Cell health declines as ergosterol and UFA reserves decline.

By pitching cells at high krausen instead of waiting until they have reached the stationary phase and prepared for starvation, we are pitching cells with non-depleted ergosterol and UFA reserves.  The difference in ergosterol and UFA reserves from pitching a yeast culture at high krausen instead of waiting until the cells enter the stationary phase results in a lower initial load being placed on dissolved O2, preserving more O2 for future generations of yeast cells. Ergosterol and UFAs make yeast cells more pliable, which, in turn, allows for the passage of nutrient and waste products through their cell walls. 

Cells with healthy ergosterol and UFA reserves are more ethanol tolerant than cells with depleted ergosterol reserves.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3274781/

"Although most organisms cannot tolerate high levels of alcohol, certain yeasts (e.g., Saccharomyces cerevisiae) are able to maintain viability in the presence of up to 15–20 vol % ethanol. Through natural and directed evolution, yeasts have developed many strategies, also known as survival factors, to deal with ethanol toxicity (1). One important survival factor is to modify plasma membrane composition by increasing the content of unsaturated lipids and ergosterol (1,2,4,5)."

The reason why yeast cells in large part stop fermenting above a certain alcohol level is because they become dehydrated causing a reduction in cell size and a loss of turgor pressure.  Ethanol is hygroscopic; hence, it draws water out of yeast cells.  A similar thing happens when we pitch yeast into high gravity wort; however, it is phenomenon known as osmotic pressure.  Osmotic pressure is the tendency of water to be drawn to the side of a semi-permeable membrane that contains the highest solute concentration.  What happens when we pitch yeast cells into high gravity wort (hypertonic environment) is that water is drawn out of the cells into the wort, which, in turn, causes cell size to decrease resulting in reduced, if not outright cessation of cellular metabolism or even cell death. 

The following paper contains images that show what happens to yeast cells when they are exposed to high gravity solutions and high alcohol levels: http://onlinelibrary.wiley.com/doi/10.1002/j.2050-0416.2003.tb00162.x/pdf

Title: Re: Shaken not Stirred
Post by: narcout on February 10, 2015, 04:59:46 PM
Sorry, spell checker erased an L in my post.  I wanted to say that I want to make two 2L starters (to pitch in two six gal. lager batches).  I don't have four 1 gal. jugs.

The shaking is just to aerate the starter wort.  You could always use an alternative method of aerating or oxygenating the starter and still achieve the benefits of pitching or crashing at high krausen.

Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 10, 2015, 05:06:50 PM
The shaking is just to aerate the starter wort.  You could always use an alternative method of aerating or oxygenating the starter and still achieve the benefits of pitching or crashing at high krausen.

That statement is correct.  However,  one cannot just flood the available headspace with O2 and shake when using an undersized container. That will not work anywhere near as well as using a larger container and normal automospheric levels of O2.  The media has to be able to expand into foam, which is the low-tech equivalent of bubbling air or O2 through the media.  The beauty of the low-tech method is that there is one less opportunity to infect one's starter.
Title: Re: Shaken not Stirred
Post by: Stevie on February 10, 2015, 05:09:29 PM
Mark, with this method, would an appropriate starter be possible with as little as two table spoons of thicker slurry?
Title: Re: Shaken not Stirred
Post by: narcout on February 10, 2015, 05:11:11 PM
The media has to be able to expand into foam, which is the low-tech equivalent of bubbling air or O2 through the media.

Yes, bubbling air or O2 through a diffusion stone is the alternative method I was referring to.

The maximum cell density for 5 gallons of wort is roughly 3.8 trillion cells; hence, pitching 400 billion cells requires log(3,800 / 400) / log(2) = ~3.25 replication periods.  Pitching 200 billion cells requires log(3,800 / 200) / log(2) = ~4.25 replication periods.

Do you think there is any discernible flavor impact from that extra replication period?
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 10, 2015, 05:48:03 PM
I need to take a photo what the amount of yeast that I pitch into 600 milliliters of wort (my primary volume is 3.5 gallons), but it is a ridiculously small amount of yeast compared to what is in a White Labs vial.  You will laugh at the thought of ever thinking that half of a White Labs vial is not enough yeast to pitch into a 1L starter when you see how little yeast I pitch.

With that said, if my math is not failing me, 2 tablespoons is just shy of 30ml. Thirty milliliters of truly thick slurry can have up to 3 billion yeast cells per milliliter, or up 90 billion yeast cells in total.  I kid you not when I say that you can pitch five gallons of ale with 90 billion fresh viable cells with no off-flavors or loss of attenuation as long as the wort is properly aerated.

Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 10, 2015, 06:03:54 PM
Do you think there is any discernible flavor impact from that extra replication period?

With proper aeration, one should not experience any discernible flavor impact from a single replication period. 

With that said, one should experience a discernible flavor impact from pitching at high krausen; namely, a cleaner tasting product.

Title: Re: Shaken not Stirred
Post by: TMX on February 10, 2015, 08:13:45 PM
If it were me, and I needed 2x2l starters, I would just order 4x1gallon jugs. I am sure you could divide the WL or Wyeast vial by 4 and still have enough yeast to make the starters.

Mark, if I am wrong here please jump in.

FWIW, these appears to be the best start to a ferment I have ever had.  The only change I wold make, is to start my starter the night prior and not 2 nights prior. I made my starters Friday night and by Sat moring they were crashing. Next time, I will start them the night prior to brewday, as most of the time I pitch around 5pm in the evening.

I ferment 10-11 gallons of wort in a 16g container, I ahve never needed a blow off tube with that much head space, when I checked this moring, the Krausen ring was within 1/2 inch of the airlock......

T
Title: Re: Shaken not Stirred
Post by: narcout on February 11, 2015, 01:45:08 AM
The following paper contains images that show what happens to yeast cells when they are exposed to high gravity solutions and high alcohol levels: http://onlinelibrary.wiley.com/doi/10.1002/j.2050-0416.2003.tb00162.x/pdf

That was an interesting read.  Did you see this part at the end?

"Yeast cells in the exponential phase had weak turgor pressure compared to turgor pressure of stationary phase yeast cells."

"Both groups postulated that the ability of a yeast strain to support a higher gradient of osmotic pressure could be due to turgor pressure and thickness of the cell membrane. Cells, which normally have high turgor, have membranes with higher tensile strength. Thus, stationary phase cells, which have been reported to be more stress tolerant than actively growing exponential phase cells, must have a more rigid cell membrane in order to maintain the same volume than exponential cells."
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 11, 2015, 03:08:12 AM
That was an interesting read.  Did you see this part at the end?

"Yeast cells in the exponential phase had weak turgor pressure compared to turgor pressure of stationary phase yeast cells."

"Both groups postulated that the ability of a yeast strain to support a higher gradient of osmotic pressure could be due to turgor pressure and thickness of the cell membrane. Cells, which normally have high turgor, have membranes with higher tensile strength. Thus, stationary phase cells, which have been reported to be more stress tolerant than actively growing exponential phase cells, must have a more rigid cell membrane in order to maintain the same volume than exponential cells."

I have mentioned that yeast cells undergo morphological changes at the end of fermentation in preparation for hard times in several thread on this forum and others. 

https://www.homebrewersassociation.org/forum/index.php?topic=21705.msg275241#msg275241

"By stepping a culture at the end of the exponential phase, we are pitching yeast cells that require very little in the way of replenishment. Hence, we will experience a shorter lag phase than we will if we pitch a culture that has reached quiescence because a quiescent culture has to undo the morphological changes it underwent in preparation for hard times.  A quiescent culture also has to replenish the ergosterol and UFA reserves that were spent post-exponential phase, which increases dissolved O2 requirements."

https://www.homebrewersassociation.org/forum/index.php?topic=21952.msg279097#msg279097

"As I have mentioned many times, a high krausen pitch will always outperform a "fermented out" pitch because the cells are in peak health and do not have to undo the morphological changes that occur at the end of fermentation.  Add in sufficient aeration and a strain with a high attenuation rate, and an AA of 85% is not out of the question."

I have also mentioned that cell wall thickening is the major morphological change.

https://www.homebrewersassociation.org/forum/index.php?topic=20692.msg262969#msg262969

"At the end of fermentation, yeast cells go into survival mode where their cell walls thicken and they store carbohydrate as glycogen.  In effect, the cells are preparing for hard times.  It takes longer to exit this state than its does when the yeast cells are still in active growth mode; hence, lag times in addition to oxygen demands are also increased."

What the authors did not mention is that this survival mechanism has to be undone during the lag phase; otherwise, nutrients and waste products cannot easily pass through the cell wall and plasma membrane.  Hence, stationary phase cells are no more likely to survive the lag phase than yeast cells that are pitched during the exponential phase, which is why we pitch at a higher rate.
Title: Re: Shaken not Stirred
Post by: Philbrew on February 11, 2015, 03:30:18 AM
Sorry, spell checker erased an L in my post.  I wanted to say that I want to make two 2L starters (to pitch in two six gal. lager batches).  I don't have four 1 gal. jugs.

Headspace is critical to achieving a proper shake.  One is attempting to turn the wort into as much foam as is humanly possible.  This method is designed to be low-tech and low-cost. 

With that said, another thing that I am attempting to dispel is the insane notion that people have to hit the cell counts provided by yeast calculators in order to have a healthy fermentation.   It is better to have 60 billion healthy, ready to go to war with the wort cells than it is to have 200 billion stressed cells that are barely clinging to life.  The difference between 60 billion cells and 200 billion cells is approximately 180 minutes of propagation time.
OK, light bulbs are turning on all over.  "as much foam as is humanly possible"  makes big time sense because foam has huge surface area. 
This engineer has always believed that "low-tech and low-cost" are the better solution. 
I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.
Thanks, Mark.  Very enlightening post on how yeast do their thing.
Title: Re: Shaken not Stirred
Post by: narcout on February 11, 2015, 04:18:05 AM
What I hadn't heard before was the part about stationary cells being reported to be more stress tolerant.

I haven't had a chance to read the article cited at the end of that sentence, but here's the link.  I have no idea if it's interesting or presents anything that you haven't already mentioned.  My guess is that you've probably read it before.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC372915/pdf/microrev00025-0099.pdf

I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.

For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 11, 2015, 05:44:57 AM
What I hadn't heard before was the part about stationary cells being reported to be more stress tolerant.

The morphological changes occur in order to harden the cells against the harsh reality of living in a toxic, low-nutrient medium for an unknown amount of time.
 
Quote
For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.


Cell count is only half of the equation.  Cell health is equally, if not more important.  When pitching cells at high krausen, one is pitching very healthy cells with very pliable cell walls that require little to no maintenance before they can go to work; therefore, reducing lag time.  The number one bogeyman with normal gravity fermentation is house infection.  Shortening the lag phase shortens the time that bacteria have to multiply (as does reducing the number of replication periods), and bacteria multiply three times faster than yeast.  These growth rates are what we are up against in a real world brewery.

https://www.homebrewersassociation.org/forum/index.php?topic=19850.msg277460#msg277460

"A small amount of bacteria can overtake a much larger amount of yeast because the bacteria cell population increases 8-fold every time the yeast cell population doubles.  If we were to normalize the propagation period between yeast and bacteria (bacteria multiplies three times faster than yeast), the growth equations would be:

yeast_cell_count = initial_cell_count * 2n, where n = elapsed time in minutes since the end of the lag phase / 90

bacteria_cell_count = initial_cell_count * 8n, where n = elapsed time in minutes since the end of the lag phase / 90


If we run the numbers, it should become crystal clear why one wants to pitch a large, healthy yeast culture while doing everything possible to minimize the opportunity for bacteria to catch a ride into one's yeast crop, starter, or fermentation vessel.  It should also become clear why the growth phase is called the exponential phase.


Cell counts at 90 minutes

yeast_cell_count = initial_yeast_cell_count * 21 =  initial_cell_count * 2
bacteria_cell_count = initial_bacteria_cell_count * 81 = initial_cell_count * 8


Cell counts at 180 minutes

yeast_cell_count = initial_yeast_cell_count * 22 =  initial_cell_count * 4
bacteria_cell_count = initial_bacteria_cell_count * 82 = initial_cell_count * 64


Cell counts at 270 minutes

yeast_cell_count = initial_yeast_cell_count * 23 =  initial_cell_count * 8
bacteria_cell_count = initial_bacteria_cell_count * 83 = initial_cell_count * 512


Cell counts at 360 minutes

yeast_cell_count = initial_yeast_cell_count * 24 =  initial_cell_count * 16
bacteria_cell_count = initial_bacteria_cell_count * 84 = initial_cell_count * 4096


Cell counts at 450 minutes

yeast_cell_count = initial_yeast_cell_count * 25 =  initial_cell_count * 32
bacteria_cell_count = initial_bacteria_cell_count * 85 = initial_cell_count * 32768


Cell counts at 540 minutes

yeast_cell_count = initial_yeast_cell_count * 26 =  initial_cell_count * 64
bacteria_cell_count = initial_bacteria_cell_count * 86 = initial_cell_count * 262,144


Cell counts at 630 minutes

yeast_cell_count = initial_yeast_cell_count * 27 =  initial_cell_count * 128
bacteria_cell_count = initial_bacteria_cell_count * 87 = initial_cell_count * 2,097,152


Cell counts at 720 minutes

yeast_cell_count = initial_yeast_cell_count * 28 =  initial_cell_count * 256
bacteria_cell_count = initial_bacteria_cell_count * 88 = initial_cell_count * 16,777,216"


Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 11, 2015, 05:47:32 AM
OK, light bulbs are turning on all over.  "as much foam as is humanly possible"  makes big time sense because foam has huge surface area. 

Exactly!
Title: Re: Shaken not Stirred
Post by: Philbrew on February 11, 2015, 07:43:09 PM

I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.

For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.
[/quote]
Sorry, I didn't mean to infer that the Mr. Malty calculator is inaccurate.  In fact it's probably the best.  It's just that all the yeast calculators seem to be overly cautious on yeast production rates except for stir plates. I realize that it would be difficult to add to the calculator the Shake-it-until-it's-all-foam method or combine O2 at start plus shaking.   But, as it is, the calculators tend to push one into buying a stir plate, especially if you want to do lagers or big beers.  I want to brew lagers and the $200 for a heavy duty stir plate and 5L flask could be better spent on a wort chiller or a temp controller for my fridge and an upgrade of my brew pot.
Title: Re: Shaken not Stirred
Post by: TMX on February 11, 2015, 07:48:25 PM

I have trouble trusting the yeast calculators that call for incredibly huge pitching rates without factoring in the state of yeast vitality.

For what  it's worth, I believe the Mr. Malty calculator uses the fairly standard rate of .75 million cells per milliliter of wort per degree Plato (and twice that for lagers). 

I'm not saying that's the be all and end all of pitching rates or that it isn't worth experimenting with different rates for different styles of beer, but it's not like Jamil just arbitrarily made it up.
Sorry, I didn't mean to infer that the Mr. Malty calculator is inaccurate.  In fact it's probably the best.  It's just that all the yeast calculators seem to be overly cautious on yeast production rates except for stir plates. I realize that it would be difficult to add to the calculator the Shake-it-until-it's-all-foam method or combine O2 at start plus shaking.   But, as it is, the calculators tend to push one into buying a stir plate, especially if you want to do lagers or big beers.  I want to brew lagers and the $200 for a heavy duty stir plate and 5L flask could be better spent on a wort chiller or a temp controller for my fridge and an upgrade of my brew pot.
[/quote]

on uline.com, you can get a 4 pack of 1 gallon glass jugs for less than $20 (plus shipping), split a single vile 4 ways, and have a 1 gallon starter.....no stirplate needed.

T
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 11, 2015, 08:16:33 PM
on uline.com, you can get a 4 pack of 1 gallon glass jugs for less than $20 (plus shipping), split a single vile 4 ways, and have a 1 gallon starter.....no stirplate needed.

Just remember to shake the jugs like they owe you money (think mafia enforcer)! :)
Title: Re: Shaken not Stirred
Post by: narcout on February 11, 2015, 08:20:27 PM
It's just that all the yeast calculators seem to be overly cautious on yeast production rates except for stir plates.

Yes, that could very well be true (or perhaps they are overly optimistic on the effects of stir plates). 

I believe at least some of the predicted results from the Mr. Malty calculator are based on real world observations, but I do not know to what extent (though this info may be out there on the internet somewhere).

Fortunately, as has been pointed out, you do not need to be exact in your pitching rates.  They are just another tool that you can use to achieve your intended flavor profile.

By the way, another good source for inexpensive one gallon glass jugs is your local supermarket (and they come filled with delicious juice).

Title: Re: Shaken not Stirred
Post by: Philbrew on February 11, 2015, 08:44:38 PM
on uline.com, you can get a 4 pack of 1 gallon glass jugs for less than $20 (plus shipping), split a single vile 4 ways, and have a 1 gallon starter.....no stirplate needed.

Just remember to shake the jugs like they owe you money (think mafia enforcer)! :)
Yup, that's my plan.  Hmmm...I wonder if my local Ace Hardware would let me put them on their paint shaker? ;)
Title: Re: Shaken not Stirred
Post by: TMX on February 11, 2015, 09:39:48 PM
I need to take a photo what the amount of yeast that I pitch into 600 milliliters of wort (my primary volume is 3.5 gallons), but it is a ridiculously small amount of yeast compared to what is in a White Labs vial.  You will laugh at the thought of ever thinking that half of a White Labs vial is not enough yeast to pitch into a 1L starter when you see how little yeast I pitch.

With that said, if my math is not failing me, 2 tablespoons is just shy of 30ml. Thirty milliliters of truly thick slurry can have up to 3 billion yeast cells per milliliter, or up 90 billion yeast cells in total.  I kid you not when I say that you can pitch five gallons of ale with 90 billion fresh viable cells with no off-flavors or loss of attenuation as long as the wort is properly aerated.

you keep threating us with this....lol

Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 11, 2015, 10:07:22 PM
you keep threating us with this....lol

I inoculate one of these media bottles with one or more 4mm loop scrapes taken from a slant. 

(http://i699.photobucket.com/albums/vv356/tonestack/Brewing/MediaBottle_zpsdff03f83.jpg)

The yeast that is grown in 40 milliliters of absolutely sterile (autoclaved) wort is pitched into 600 milliliters of 10% w/v (1.040) wort.  That's a one to fifteen step media-wise, which is living on the edge when dealing with such a small amount of yeast.   A White Labs vial contains almost 35ml of yeast cells.  I am pitching at best 1/10th that amount.
Title: Re: Shaken not Stirred
Post by: TMX on February 11, 2015, 10:22:47 PM
i need to see if they have autoclaves on Craigslist
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 12, 2015, 01:29:49 AM
i need to see if they have autoclaves on Craigslist

You do not need to purchase a laboratory autoclave.  A pressure cooker/canner that is capable of 15 PSI above normal atmospheric pressure will get the job done.  You need to be careful when purchasing because many modern pressure cookers/canners are not capable of achieving 15 PSI above normal atmospheric pressure pressure.  All of the "All American" pressure cookers/canners are capable of achieving this pressure level.  A few of the lower-priced pressure cookers are capable of achieving 15 PSI above normal atmospheric pressure as well.
Title: Re: Shaken not Stirred
Post by: TMX on February 12, 2015, 04:49:18 AM
This has been sitting at 66 since Sunday night, I have to leave town Fri-Mon, would there be any benefit it raising the temp up to 70ish while I am out of town
Title: Re: Shaken not Stirred
Post by: S. cerevisiae on February 12, 2015, 06:37:02 PM
If you want to raise the temperature, go ahead and do it.  I am a "steady state" brewer when it comes to ale.  However, then again, I brew mostly British-style ale when I brew ale.
Title: Re: Shaken not Stirred
Post by: TMX on March 06, 2015, 01:20:12 PM
On my 3rd set of starters using the "James Bond" method.

At some point a detailed write up should be a sticky.

here is my 1056 at 10 hours!

(http://i42.photobucket.com/albums/e346/TXAleWorks/Yeast/IMG_1174_zpsqua0ocvj.jpg)
Title: Re: Shaken not Stirred
Post by: quattlebaum on March 17, 2015, 03:41:08 AM
Do you think there is any discernible flavor impact from that extra replication period?

With proper aeration, one should not experience any discernible flavor impact from a single replication period. 

With that said, one should experience a discernible flavor impact from pitching at high krausen; namely, a cleaner tasting product.

I know this is an older thread but i have been trying to get a cleaner and less phenolic character from Wyeast 3068 and have been fermenting at 62F with a suggested starter from one the yeast cal.  Wondering if i would get a "cleaner" profile from this strain if i pitched at high krausen? Interesting for sure maybe i will try it.
Title: Re: Shaken not Stirred
Post by: TMX on March 17, 2015, 09:29:38 PM
best way to find out is to try!
Title: Re: Shaken not Stirred
Post by: JT on March 17, 2015, 10:01:10 PM
Do you think there is any discernible flavor impact from that extra replication period?

With proper aeration, one should not experience any discernible flavor impact from a single replication period. 

With that said, one should experience a discernible flavor impact from pitching at high krausen; namely, a cleaner tasting product.

I know this is an older thread but i have been trying to get a cleaner and less phenolic character from Wyeast 3068 and have been fermenting at 62F with a suggested starter from one the yeast cal.  Wondering if i would get a "cleaner" profile from this strain if i pitched at high krausen? Interesting for sure maybe i will try it.
I think hefe phenolics have more to do with pitch rate than pitching at peak activity... unless the transition from being pitched at high krausen also causes less stress - that indeed may be beneficial.  I think wyeast recommends over-pitching to cut down on banana.  Ensuring it is well aerated/oxygenated may also limit ester production and if you can ferment under more pressure,  that would likely help as well. 
Title: Re: Shaken not Stirred
Post by: johnnyb on July 07, 2015, 07:43:12 PM
I'm going to try this (with oxygen instead of shake n' bake) tomorrow for a beer I'm hoping to brew Sunday and have a couple of questions.

How many seconds of pure O2 with a 2 micron stone into 1 liter?

How do you know when you're completely at high krausen without going past it? Is it better to crash a little sooner or a little later?

On brew day, should I decant and then let the slurry warm up prior to pitching? Or take out of fridge, decant, and pitch still cold? (In the past, I've always done the latter but not sure it's correct.)

Thanks for any comments!


Edit: actually my stone might be 0.5 micron -- I can't remember. It's stainless on a wand. I got it at Williams several years ago, but they no longer have a record of the transaction in my account. Don't know if it matters for the bubbling time? In 5.5 gallons I always bubble for 45 to 60 seconds.
Title: Re: Shaken not Stirred
Post by: JT on July 08, 2015, 10:46:36 AM
I'm going to try this (with oxygen instead of shake n' bake) tomorrow for a beer I'm hoping to brew Sunday and have a couple of questions.

How many seconds of pure O2 with a 2 micron stone into 1 liter?

How do you know when you're completely at high krausen without going past it? Is it better to crash a little sooner or a little later?

On brew day, should I decant and then let the slurry warm up prior to pitching? Or take out of fridge, decant, and pitch still cold? (In the past, I've always done the latter but not sure it's correct.)

Thanks for any comments!


Edit: actually my stone might be 0.5 micron -- I can't remember. It's stainless on a wand. I got it at Williams several years ago, but they no longer have a record of the transaction in my account. Don't know if it matters for the bubbling time? In 5.5 gallons I always bubble for 45 to 60 seconds.
One problem pitching cold can be getting the yeast off the bottom of your container, certain strains are harder than others.  I let it warm up a bit first.  Don't stress about crashing right at high krausen, if it goes over IME you'll be just fine.  There are many people let their yeast sit on a stir plate for days (I'm not recommending) and make good beer - decanting the starter becomes more important though. 
As for your o2 time, mathematically it is around 3 seconds per liter.  I did mine for 10.  No worries.  You'll still want to shake the hell out of it. 

Sent from my SCH-I545 using Tapatalk
Title: Re: Shaken not Stirred
Post by: johnnyb on July 08, 2015, 10:45:00 PM
Thanks!

I picked up a pack of Denny's 50 this morning with a June 25 manufacture date. Smacked it about noon. Pitched into the starter at 2pm. It started bubbling around 5:30 and is getting more frequent now. No krausen yet, but I can tell it's close to forming.


Edit: times are EST.
Title: Re: Shaken not Stirred
Post by: johnnyb on July 09, 2015, 04:44:58 AM
Well it's been almost 11 hours and I never got a krausen, but it seems to obviously be past peak and yeast is starting to settle out on the bottom so I think I'm going to crash it now.
Title: Re: Shaken not Stirred
Post by: johnnyb on July 12, 2015, 01:42:46 AM
Not quite 5 hours after pitching this starter in my wort and I'm seeing the first signs of take off!

Edit: Full blast off with 2" krausen in sometime less than 13 hours. (Not sure exactly when because I was sleeping.)