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bodhisatta 
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Blue Helix]
#26999654 - 10/23/20 12:34 PM (3 years, 3 months ago) |
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Liquid cultures are not battle zones. Many different organisms will live together and not outright kill each other off. Anytime beer gets contaminated bacteria have to cope with yeast's defenses, the low pH, the hop acids. And yet they still persist along side the intended microorganisms if they managed to get in
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Blue Helix
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: bodhisatta]
#27000137 - 10/23/20 05:35 PM (3 years, 3 months ago) |
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Quote:
bodhisatta said: Liquid cultures are not battle zones. Many different organisms will live together and not outright kill each other off. Anytime beer gets contaminated bacteria have to cope with yeast's defenses, the low pH, the hop acids. And yet they still persist along side the intended microorganisms if they managed to get in
I have opened LCs that smell obviously of bacteria, and I even fruited them out. But the yields suffer. If you open one that only smells of malt sugar and mycelium, the grows are usually great. I'm not saying there always is a victor, but when there is, it works a lot better. I generally find that the more times I propagate an LC, the faster it develops and the less likely it'll smell of anything but light malt and mycelium after opening the LC jar. I sometimes wish I could just smell the LC without contaminating it because you can tell so much from the smell, but I don't know how to do that. Let me know if you do.
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Blue Helix
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Blue Helix] 2
#27000367 - 10/23/20 07:49 PM (3 years, 3 months ago) |
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Let me just put it this way: I believe most LCs do not establish an equilibrium with bacteria. Here are the three types I've seen settle out:
Contamination 1) The LC will become more cloudy every day until you can barely see in it. Allowing the LC to settle overnight does not resolve the cloudiness. 2) The mycelium, if you get any at all, will start to seem constricted in growth (tight spheres with no obvious growth in the number of size of them) 3) The mycelium will eventually die, the blobs collapse becoming dead strings in the LC 4) Upon opening the jar, it'll smell very bad
Ideal (just mycelium) 1) The mycelium will grow exponentially at first sometimes even becoming like a slush before stopping 2) The LC will become crystal clear between the mycelium blobs when spinning or if you let it settle overnight 3) When you open it, you smell mostly just malt (if you use that) and mycelium. A very faint bad smell is still possible but it should not be overpowering
Some kind of equilibrium between mycelium, bacteria, and/or yeast 1) My mycelium will grow but not very fast 2) The LC will stay cloudy but not get more cloudy 3) When you open it, the LC will smell funky but not overpoweringly so. In fact sometimes it just smells weird more than bad
The first type is a complete failure and should be disposed. The second is what you want and usually will get.
The third type of LC is very rare (<1 in 100). I believe the mycelium is in some kind of equilibrium with the bacteria and/or yeast, but that does not help with yield. Also in my experience per-flush yield practically flatlines after the first flush. One way to clean it up would be this agar sandwich technique. Another way is just to expand the LC a time or two. Take say a ml of it and expand it out to 500ml one or two times. When I've done that in the past, usually the mycelium tends to take over, and the cloudiness vanish.
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Blue Helix
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Blue Helix]
#27001248 - 10/24/20 01:03 PM (3 years, 3 months ago) |
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So the scales of bacteria versus mycelium in the LC finally tipped last night toward mycelium. This morning, the LC has flocculated the malt solids into small mycelium balls. When this happens, the LC never goes back to bacterial in my experience. I might expand it out one more time just to give it a totally fresh start, but that's not necessary. This is what an LC looks like the day it flips:

The size of the balls is so small because there were a lot of malt solids in this LC. They had been ground up through the ordeal, so this is typical of that. Although it's kind of hard to tell from the animated GIF, the cloudiness between the mycelium pieces is gone. That's because the mycelium sequestered all the malt solids and bacteria into the balls.
If I open this, it might still have some bacterial smell, but that's because it was on the edge and fighting just days ago. You cannot expect all that bacterial smell to just vanish, but the bacteria doesn't have the upper hand here. A fresh LC inoculated with this will not have a bacterial smell and the balls will be larger.
So this is proof that a culture that I failed to expand 4 times previously due to bacteria could be saved using an agar sandwich technique as outlined elsewhere on the site. I had to see it for my own eyes to believe it, but this LC makes it undeniable. The agar sandwich process did clean up the culture.
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bodhisatta 
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Blue Helix]
#27001375 - 10/24/20 02:42 PM (3 years, 3 months ago) |
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Dude have you ever heard of kombucha. Look up kombucha and scobys
You can have your beliefs but even louis pasteur could tell you your LC theory is ridiculous at best. A cheapo microscope is like $120 you'll see what you call very rare is the most common
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Blue Helix
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: bodhisatta]
#27001434 - 10/24/20 03:16 PM (3 years, 3 months ago) |
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bodhisatta said: Dude have you ever heard of kombucha. Look up kombucha and scobys
You can have your beliefs but even louis pasteur could tell you your LC theory is ridiculous at best. A cheapo microscope is like $120 you'll see what you call very rare is the most common
You are kidding me, right? Please tell me this is a joke. I'm an LC guy for 20 years and you wonder if I've heard of the stable relationship between bacteria and yeast we call kombucha? Come on, man! Get real. And do YOU know why it's unusual and something to take note? That would be because it is stable and doesn't kill you when you drink it. Also what's missing in the kombucha? Oh, yeah, mycelium.
My guess is that you are one of these beer brewer guys who thinks because he can brew a batch of beer that he knows all there is to know about liquid cultures. The only thing I know about beer brewing is that having known people who make them, they are ridiculously easy because you can be a slob and not really worry about infecting them (kind of like kombucha which is also easy for any slob to make). Brewing is a different world - one where sloppy work hardly matters because everything is trivial to keep clean.
Look you are obviously trolling me, which is kind of shocking for a guy with 58,000 posts. Back when I was the moderator for the advanced cultivation forum, we didn't have these trolling issues. The massive trolling seems relatively new - as in the last decade.
Anyway, I promised the moderators to stop entertaining trolls, so I have to shut up. However, anyone reading this who does have information that suggests that an LC does in fact establish an equilibrium of bacteria and mycelium typically, please write me. I'll be most interested in having the mycologist such as Alan Rockefeller show me where those sneaky bacterium are hiding in my LC. I just don't seem to be able to find them. Maybe they are hiding... Nah, I bet they aren't there.
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bodhisatta 
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Blue Helix]
#27001437 - 10/24/20 03:18 PM (3 years, 3 months ago) |
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 You're clueless. Beer is retarded easy to infect. I can see your demeanor in calling me a troll for pointing something out to you so it's absolutely no use trying to have any logical or fruitful conversation so peace out and good luck
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Blue Helix
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: bodhisatta]
#27003787 - 10/25/20 10:37 PM (3 years, 3 months ago) |
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Just to emphasize what a LC that is already going is like, this LC was started about 36 hours ago from the 25ml of the clean one above. As you can see, it's within a day of being plenty thick to use (actually it could be used now really even while it still needs a good 6 to 12 hours to flocculate the malt solids which will clear it up).

Also when I open this one, there will be no bacterial smell, and that's because once you expand a good clear LC, bacteria is completely gone. There is zero bacteria as can easily be established via microscope or agar plate (I thank Alan Rockefeller for his tips about using a syringe filter to make sure). It'll never smell bacterial because it's not there, which was the whole point of this exercise.
So I expanded 25 to 500ml within a day and a half, but you could just as easily expand and 500ml LC into several gallons in two days if you wanted. It's really endless, which is one reason I like LCs; they are fast.
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Kizzle
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Blue Helix]
#27003961 - 10/26/20 02:43 AM (3 years, 3 months ago) |
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I have a couple questions: first, have others had luck with these agar sandwich techniques and do you have any pointers for doing it? And, secondly, why would bacteria not overwhelm mycelium on an agar plate yet do so in a liquid culture? What is it about liquid cultures that tip that balance so much toward bacteria? Microscopically what is going on?
This is agar seen with an electron microscope. It's essentially an LC except the agar structure holds everything in place so it's more like a solid. Molecules can diffuse in it but bacteria are too large and grow mostly on the surface. Without the agar the bacteria in a LC can quickly spread to every bit of the liquid medium which is something mycelium can't do as well because the cells are connected.

I'm not sure if the mycelium can actually digest the agar but the way mycelium grows it can force it's way through it.
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Edited by Kizzle (10/26/20 02:48 AM)
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Blue Helix
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Re: Relationship of bacteria and mycelium in/on various nutrient media [Re: Kizzle]
#27004013 - 10/26/20 05:09 AM (3 years, 3 months ago) |
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Quote:
Kizzle said:
Quote:
I have a couple questions: first, have others had luck with these agar sandwich techniques and do you have any pointers for doing it? And, secondly, why would bacteria not overwhelm mycelium on an agar plate yet do so in a liquid culture? What is it about liquid cultures that tip that balance so much toward bacteria? Microscopically what is going on?
This is agar seen with an electron microscope. It's essentially an LC except the agar structure holds everything in place so it's more like a solid. Molecules can diffuse in it but bacteria are too large and grow mostly on the surface. Without the agar the bacteria in a LC can quickly spread to every bit of the liquid medium which is something mycelium can't do as well because the cells are connected.
I'm not sure if the mycelium can actually digest the agar but the way mycelium grows it can force it's way through it.
Sounds exactly like what I said - that bacteria's motility in LC is basically infinite. It also provides the solid footing of evidence that an LC eventually scours itself of bacteria when it clears up. If it didn't do that, it would continue to smell bad, be cloudy, etc. Lastly, it describes how it is that bacteria can be removed by forcing the mycelium to grow through the agar. Thank you!
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