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Laboratory propegation and Brewery propegation of yeast (SOPs)

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  • Yeast
    replied
    Originally posted by Swags View Post
    Thanks for your response.

    Our tank has a 6" TC opening on the top so my plan is to pour the 4 liters in through that opening. We'll try the flame method you showed in your photos.

    Also now considering running a small march pump as a recirculation method for a day or two after aerating for 24 hours.
    best of luck. It wouldn't be a bad idea to wear some gloves and a "I have the flu" face mask for this task. having a torch running in the area is a good idea just be careful about its location, don't melt anything. Even plastic bottles can have their openings flamed briefly.

    Leave a comment:


  • Swags
    replied
    Propagating from Yeast Lab Pitch

    Thanks for your response.

    Our tank has a 6" TC opening on the top so my plan is to pour the 4 liters in through that opening. We'll try the flame method you showed in your photos.

    Also now considering running a small march pump as a recirculation method for a day or two after aerating for 24 hours.

    Leave a comment:


  • Yeast
    replied
    Originally posted by Swags View Post
    Yeast,

    Fabulous post!

    We are a smallish brewery with fairly large fermentation tanks so we want to grow up our pitches that we buy locally. We do not have a lab nor time available to propagate using flasks. Basically, we want to skip the beginning steps of your SOPs and get right into the 3 bbl propagator.

    We just acquired a 200 gal. tank that we want to dedicate as a yeast propagation vessel. We have HEPA air filters and will attach an aeration stone to the bottom outlet.

    Question: If we buy industry standard pitches from our local yeast lab at 7 mill. cells / ml and pitch 4 liters of that into 100 gallons of 12 P. wort (with aeration), can we expect the same 10 fold increase in cell count that you got from your higher density 4 L pitches? If not, what kind of cell density can we expect using these types of pitches?

    Thanks for any input!
    Two issues right off the bat.
    Can you trust the standard pitches you buy? Propagation was brought to the table to get away from problems with bought pitches.

    The other problem is inoculation of the prop tank with the bought pitch. Good luck achieving a completely aseptic transfer of all the bought yeast into the prop.
    At least the prop is aerobic so you won't necessarily grow anaerobic organisms, but wild yeast or even other brewery yeast can become an issue.
    This is why I went through the hassle of the lab side prop steps from dishes to flasks.

    Otherwise your plan checks out.

    Leave a comment:


  • Swags
    replied
    Propagating from Yeast Lab Pitch industry standard

    Yeast,

    Fabulous post!

    We are a smallish brewery with fairly large fermentation tanks so we want to grow up our pitches that we buy locally. We do not have a lab nor time available to propagate using flasks. Basically, we want to skip the beginning steps of your SOPs and get right into the 3 bbl propagator.

    We just acquired a 200 gal. tank that we want to dedicate as a yeast propagation vessel. We have HEPA air filters and will attach an aeration stone to the bottom outlet.

    Question: If we buy industry standard pitches from our local yeast lab at 7 mill. cells / ml and pitch 4 liters of that into 100 gallons of 12 P. wort (with aeration), can we expect the same 10 fold increase in cell count that you got from your higher density 4 L pitches? If not, what kind of cell density can we expect using these types of pitches?

    Thanks for any input!

    Leave a comment:


  • Yeast
    replied
    Originally posted by gagarin View Post
    Why don't you autoclave wort in Erlenmayer flasks directly?
    To leave the trub behind in the media bottles

    Leave a comment:


  • gagarin
    replied
    Originally posted by Yeast View Post
    500mL media bottles were used to collect and autoclave the wort. the flasks were autoclaved separately with their stoppers on a dry cycle. then the sterilized wort is transferred into the flasks leave the trub behind
    Why don't you autoclave wort in Erlenmayer flasks directly?

    Leave a comment:


  • Yeast
    replied
    Originally posted by mobcraftbeer View Post
    I am looking at bringing yeast propagation in house, and have been doing quite a bit of research over the last 2 weeks, so i appreciate the post, but have a few questions:

    1. At what point(s) do you collect prop samples for micro analysis before pitching?

    2. What Media do you use to look for spoilage organisms during propogation phase and what are typical incubation times?

    3. I am considering supplementing olive oil (in addition to shaker tables) into wort to help with yeast building up cell mass during early stage of prop (10ml-10L), but was wondering if that would cause the yeast to loose some of the genes early on for aerobic respiration (not familiar with yeast genetics enough to know unfortunately) and was wondering if you or anyone else on this forum had experimented around with it. Link to article explaining this here:

    https://byo.com/malt/item/1206-olive-oil-aeration

    4. How many generations are you able to get out of your yeast propogating it this way and how often do you reculture your slants/yeast sources?
    1. really any time possible after the first day.

    2. BMB(mike barney miller agar) and UBA as well as LWYM (3711 french saison grows on this BTW)

    tests take 7 days. PCR is expensive then you need good primers anyway. So it would be known after the fact if something went wrong, but at least you can stop it before packaging which is a huge loss

    3. I've heard of the sterol oil treatment actually saw it first in a paper by New Belgium. From what I've read on the oil/aeration thing it's best to continue to aerate and avoid the oil. The patco anti-foam that's hydrogenated oil based not silicone apparently helps with the cell wall sterols and I suggest the use of anti foam in the prop anyway.

    4. realistically about 10 the big problem was brew scheduling and being able to juggle it into active fermentations then harvest it without it sitting around waiting for more than a day or two. What killed the yeast being used 10+ times wasn't related to quality control most of the time.
    honestly I would get LHBS vials/smack packs and make my own streak plates then isolate colonies that way. You would be surprised what contaminants grow on the plates you streak from a home brew vial on. I also didn't have access to a flow hood so sterile technique was a bit relying on ancient technology

    The other brewery I worked at would push yeast 20-30+ generations and didn't propagate at all. But they also had a very efficient and predictable brew schedule and was able to harvest yeast completely aseptic with no exposure to the atmosphere at any point.

    Leave a comment:


  • UnFermentable
    replied
    Originally posted by mobcraftbeer View Post
    At what point(s) do you collect prop samples for micro analysis before pitching?

    What Media do you use to look for spoilage organisms during propogation phase and what are typical incubation times?

    How many generations are you able to get out of your yeast propagating it this way and how often do you re-culture your slants/yeast sources?
    These were the only points I would suggest to include, but a very good write up overall.

    I would suggest number of generations is limited only by viability, and fermentation characteristics? It sounds like you use a lot of strains, so I would guess you typically don't stretch the generations to their full potential? Probably make slurry more often, since you have the ability?

    Again, nice write up.

    Leave a comment:


  • mobcraftbeer
    replied
    A few questions

    I am looking at bringing yeast propagation in house, and have been doing quite a bit of research over the last 2 weeks, so i appreciate the post, but have a few questions:

    At what point(s) do you collect prop samples for micro analysis before pitching?

    What Media do you use to look for spoilage organisms during propogation phase and what are typical incubation times?

    I am considering supplementing olive oil (in addition to shaker tables) into wort to help with yeast building up cell mass during early stage of prop (10ml-10L), but was wondering if that would cause the yeast to loose some of the genes early on for aerobic respiration (not familiar with yeast genetics enough to know unfortunately) and was wondering if you or anyone else on this forum had experimented around with it. Link to article explaining this here:

    https://byo.com/malt/item/1206-olive-oil-aeration

    How many generations are you able to get out of your yeast propogating it this way and how often do you reculture your slants/yeast sources?

    Leave a comment:


  • gculhane
    replied
    Thanks for Sharing it !!!!!

    Very Well Written, Good Information, Thanks for Sharing it !!!!!
    Gregg
    Stockton

    Leave a comment:


  • Yeast
    replied
    Originally posted by chinsi View Post
    Thank you so much for taking the time out to put this together! We're looking to start propagating in house for cost/availability reasons and I'm tasked with it. We're only looking to do a 10 HL pitch so even a smaller vessel would suffice. I am fairly comfortable doing propagation with flasks so one way we can achieve this right now is to ferment 'half' a batch and then use the harvested yeast from that for successive batches. However from a production stand point it's a little inefficient and it might make more sense for us to get a smaller propagation vessel instead of locking up a large conical. I would like to get a smaller ~2 HL dedicated vessel for yeast prop but I am still trying to wrap my head around the logistics of it, aeration being one of my biggest concerns. Does simply bubbling pure, filtered O2 through the dump port work well or will a sintered diffusion stone be necessary? I was thinking of using one of these and modifying them.
    I had simply bubbled filtered AIR through the bottom. I wouldn't use O2 unless you can measure it very accurately
    "The regulator goes to an airline that pumps through the bottom cone for agitation and air to keep the yeast aerobic. I find that 10PSI works well for this.

    I also keep the regulator above the level of the liquid in the propegator, even though there's an inline check valve, this is an additional measure to prevent any backflow to the filter or regulator"

    Leave a comment:


  • chinsi
    replied
    Thank you so much for taking the time out to put this together! We're looking to start propagating in house for cost/availability reasons and I'm tasked with it. We're only looking to do a 10 HL pitch so even a smaller vessel would suffice. I am fairly comfortable doing propagation with flasks so one way we can achieve this right now is to ferment 'half' a batch and then use the harvested yeast from that for successive batches. However from a production stand point it's a little inefficient and it might make more sense for us to get a smaller propagation vessel instead of locking up a large conical. I would like to get a smaller ~2 HL dedicated vessel for yeast prop but I am still trying to wrap my head around the logistics of it, aeration being one of my biggest concerns. Does simply bubbling pure, filtered O2 through the dump port work well or will a sintered diffusion stone be necessary? I was thinking of using one of these and modifying them.

    Leave a comment:


  • Yeast
    replied
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    here's all 3 sizes of flasks 250ml 1L and 4L

    the 250ml are ready to be transferred to the 1L

    and I'm going to combine the four 4L into just two 4L flasks since I don't have the room in the incubator shaker.

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    the 250mL flasks have lager yeast in them right now it's extremely flocculant and as soon as you stop agitation it starts to floc out.

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    all 3 250mL flasks transferred into the 1L flasks


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    the four 4L flasks had 1L each in them. There's no room on the shaker for all these flasks so the big ones were consolidates so there is now two 4L flasks with 2L each in them of Wyeast 3711 which grows to 250 million cells in propegation easily. the lager yeast usually hits 175-200.

    Leave a comment:


  • Yeast
    replied
    Originally posted by Fausto Yu-Shan View Post
    Attached is the wort agar description, sorry it is in German.

    [ATTACH]31720[/ATTACH]



    In the USA you can obtain the wort agar from the Brewing Science Institute.

    http://www.brewingscience.com/
    Mein Deutsch ist sehr scheiss zu sprechen.. Aber wenn es lesen ist, kann ich ein bisschen besser.

    Maybe that made no sense. It's been years...

    I've seen BSI's wort agar before. I guess it didn't cross my mind you were talking about the product rather than the home made stuff

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    this GW kent 3bbl set up as a propegator can achieve some pretty thick slurries when crashed to the mid 40F range. 1272 recently yielded 80L of 8.85x10^8th slurry with the propegator filled to 3.5bbl (with anti foam) of ~10 plato wort
    Last edited by Yeast; 03-20-2016, 09:56 PM.

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  • Fausto Yu-Shan
    replied
    Wort Agar (W├╝rzeagar) description

    Attached is the wort agar description, sorry it is in German.

    Wurze Agar 356e.pdf



    In the USA you can obtain the wort agar from the Brewing Science Institute.

    http://www.brewingscience.com/
    Last edited by Fausto Yu-Shan; 03-20-2016, 09:19 PM.

    Leave a comment:

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