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OfflineBlue Helix
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Re: Some notes about LCs [Re: verum subsequentis]
    #26984526 - 10/14/20 05:37 AM (3 years, 3 months ago)

So I have a confession: I found out that the tiny bit of mycelium dying first spin might not have been what was happening.  Instead I think it was bacteria and I tricked myself into believing it was only the spinner.  I found out about this weakness of agar to hide many types of bacteria in this thread.  I am finally getting a handle on why it is that agar people need to make so many transfers and why their spawn made from an agar wedge bulk yields aren't that great a lot of times. 

And it boils down to this: it can be VERY tricky to spot bacteria on a plate, and the plate can easily fully-colonize with bacteria riding on the mycelium.  There are techniques to get rid of it, but if you don't do that and create spawn with mycelium that had bacteria riding it, your substrate might look fine but not yield well.  I hadn't known that agar can so well hide bacteria.  LCs, on the other hand, let you know if there is bacteria easily because (a) they will be clear if totally clean and (b) they will both be cloudy and smell off when dirty.  There is less of this no-man's land in an LC where things are unknown than there is on an agar dish, so you are given very powerful signal telling you if the LC has resolved all its bacterial issues or not, even if it had them earlier. On the other hand, if an LC does have bacteria all you can do is wait to see if it resolves by itself, and if you use it cloudy, it might colonize the bulk and even fruit but will have a poor pin set or no mushrooms at all.  I'm learning in this hobby all the time!

So I am once again amazed by the superiority of LCs over agar in identifying a strong, clean culture, so taking so much of the guess work away from the grower to make really big yields!  Now I am starting to understand why I've had so much luck with LCs; they help the grower know when things are just right.

Below is a picture set of how an LC can often clear up. In the last picture, it is ready to use, and the yield was great.



to



to




Edited by Blue Helix (10/14/20 05:51 AM)


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OfflineMycfunkd
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Re: Some notes about LCs [Re: Blue Helix]
    #26984676 - 10/14/20 08:17 AM (3 years, 3 months ago)

:takingnotes:


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Offlinetrippleblack
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Re: Some notes about LCs [Re: Mycfunkd]
    #26984694 - 10/14/20 08:30 AM (3 years, 3 months ago)

^are you saying the lc ate the bacteria?

almost makes we want to break out the microscope and get into antibacterial agar..


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Re: Some notes about LCs [Re: trippleblack]
    #26984914 - 10/14/20 11:15 AM (3 years, 3 months ago)

I’ve been saying for years that bacteria is the cause of lower yields. It won’t always result in a moldy tub but it often will cut your yields.


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Offlineverum subsequentis
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Re: Some notes about LCs [Re: Pastywhyte]
    #26984921 - 10/14/20 11:21 AM (3 years, 3 months ago)

Quote:

and I tricked myself into believing...




That'll happen.


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OfflineBlue Helix
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Re: Some notes about LCs [Re: trippleblack]
    #26984938 - 10/14/20 11:26 AM (3 years, 3 months ago)

Quote:

trippleblack said:
^are you saying the lc ate the bacteria?

almost makes we want to break out the microscope and get into antibacterial agar..




Mycelium and bacteria competing for resources happens all the time, LC or agar or bulk substrate.  The two are locked in an eternal battle in nature as well.  The questions become which substrates offer a competitive advantage to bacteria and which medium make it clear that either bacteria or mycelium have turned out victorious?  A great example of all this is the old example PF Tek.  We know that PF Tek works, but in my experience, most spore syringes are so filthy with bacteria that they will not even colonize an agar plate without not only a battle ensuing but with one that the bacteria wins.  And if you use them in an LC - usually the mycelium ends up victorious.  On a PK Tek brick most of the time promotes mycelium to be victorious and eventual casing-less fruiting does happen.

What I'm saying here are two things.  One, some medium promote one over the other to win, and two, some mediums make it clear when one has won the other whereas others it's harder to tell.  If you take the case of agar versus LC, a trained eye can tell on both, but with an LC it's a whole lot easier to tell that bacteria has won out because the LC goes clear.  On agar, it's a matter of the uniformity of the growth like that thread I posted here talks about, and it's much harder to tell.  It doesn't surprise me then why LCs perform so well and why people are often complaining about them.  Many people, for example, are spinning their LCs way too fast because they are using junk spinners from China, and those don't allow one to easily see when the mycelium wins out.  They blend the whole mess up too fast for that.  Then someone uses the LC before its all mycelium and wonder why they either don't get fruiting or get weak fruiting.


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Blue Helix]
    #26984968 - 10/14/20 11:42 AM (3 years, 3 months ago)

One medium I am not very familiar with are pure grain jars like what is used for spawning.  My guess is that they cannot hide bacteria well, but I don't know that for sure since I don't use them.  The bulk bags I use here, though, certainly can hide it well.  They hide it as well as those PF Tek bricks, but if you case them you won't get great results from a bag that has not resolved its growth.  So in a way, I think grain jars (for spawn) are a kind of check against the problems I am seeing, but the bulk bags I use here aren't as much.


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Re: Some notes about LCs [Re: Blue Helix]
    #26985027 - 10/14/20 12:17 PM (3 years, 3 months ago)

Bacteria can be sneaky in grains for sure. They have so much more R value than something like a bulk substrate media like manure, that it takes a lot longer for the heat to penetrate the centres of the vessels. Water has a super low R value and so any media saturated with it will come up to temp far quicker.


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Offlinetrippleblack
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Re: Some notes about LCs [Re: Blue Helix]
    #26985037 - 10/14/20 12:21 PM (3 years, 3 months ago)

Great insight Lots of good info.. understood..

i spin my jars as fast as she can go.. i assume thats one of the reasons i don't see contamination in lc; but have seen it appear once tested on agar.


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OfflineBlue Helix
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Re: Some notes about LCs [Re: trippleblack]
    #26985753 - 10/14/20 06:26 PM (3 years, 3 months ago)

Quote:

trippleblack said:
Great insight Lots of good info.. understood..

i spin my jars as fast as she can go.. i assume thats one of the reasons i don't see contamination in lc; but have seen it appear once tested on agar.




You won't see it because you CANNOT see it.  If you blend your LCs so fast that they homogenize, you'll never see the cloudiness unless you let it settle.  That is a hazard too.  It's one reason why I don't think spinning them as fast as they can go is a great idea, and I've gone into length about this in other threads.

I recommend spinning them under 200 RPM.  This is impossible with cheap ones or ones made from PC fans because those always spin up super fast.  Spinning nice and slower does a couple things.  First, it allows you to still see the cloudiness if you have bacteria contamination.  Secondly, it allows one to see the shape of the mycelium blobs in the case of mold.  Mold mycelium looks nothing like what we want in a slowly-spun LC and mold will not form discrete larger balls (it can't).  So by spinning it fast you basically can't see mold or bacteria because everything will just look the same.  If you don't have either one, then I guess it doesn't matter.  But since everyone seems to complain endlessly how LCs hide contamination, I know the reason is they cannot see it is that they spin them too fast to see it.  If you spin them slowly, you can see contamination easily.


Edited by Blue Helix (10/14/20 06:35 PM)


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Pastywhyte]
    #26985790 - 10/14/20 06:49 PM (3 years, 3 months ago)

Quote:

Pastywhyte said:
I’ve been saying for years that bacteria is the cause of lower yields. It won’t always result in a moldy tub but it often will cut your yields.




Yes, and I'm slowly learning that too.  It's taken me awhile, but I'm starting to get that bacteria doesn't work like I thought.  I always used to think it's so cut-and-dry with bacteria.  Either you get mushrooms or you get bacteria.  Then I started to question that when I noticed that if I made 10-pound bags with a known-good LC that the bags would stall.  If I inoculated 7-ound bags with the exact same LC, they'd go great.  The reason was that the cooker was too packed and 5 hours just wasn't enough time for 20-pounds(2 x 10-pound bags).  The whacky thing was, though, that when you'd open the bags after they stalled, there was no off smell.  It just smelled like earth.  So that made me realize how bacteria can hide in bulk substrates.

Then I started to see this problem with the LCs.  The agar plate looked fine, so I couldn't figure it out.  It's just that bacteria can be sneaky.  It can lay dormant on mycelium in a perfectly decent looking plate and when the conditions are right, it can cloud up your LC and ruin  it. 

The point is that it is not black-or-white with bacteria.  There is a lot of sneaky stuff that can happen that can really mess up a grow's potential.


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Re: Some notes about LCs [Re: Blue Helix]
    #26986777 - 10/15/20 10:27 AM (3 years, 3 months ago)

guys I need more notes.


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OfflineBlue Helix
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Re: Some notes about LCs [Re: mushhead] * 1
    #26988730 - 10/16/20 03:55 PM (3 years, 3 months ago)

Quote:

mushhead said:
guys I need more notes.




You mean with reference to the dead spots in the trays?  I sometimes have that too, and I found it was because of the poor casing colonization (usually over colonized but under too).  I'd recommend that you work on your casing colonization.  It's too far colonized and bordering on overlaid, so the flush is sparse much like a tray without a casing.  With proper casing colonization, the flush will be about twice that dense.  Here is an example of a flush with proper casing colonization versus two without:

Ideal casing colonization:


Over casing colonization (and typical of no casing at all):


Under casing colonization:


All trays were LC trays, but in both cases of over and under casing colonization, you can see the reduced pin set.  The only problem with perfect casing colonization is that it is kind of hard for me to get consistently.  Even now I usually don't get it right, and I don't know why.  It seems to have something to do with the timeframe that the substrate transitions into rhizomorphic growth and could also be related to concurrent bacteria levels in the substrate. There might be a genetic component too.


Edited by Blue Helix (10/16/20 04:03 PM)


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InvisibleMateja
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Re: Some notes about LCs [Re: Blue Helix]
    #26989521 - 10/17/20 03:36 AM (3 years, 3 months ago)

Quote:

Blue Helix said:
in my experience, most spore syringes are so filthy with bacteria that they will not even colonize an agar plate without not only a battle ensuing but with one that the bacteria wins.  And if you use them in an LC - usually the mycelium ends up victorious.


 
Are you saying that filthy spore solutions that are unable to germinate healthy myc on plates  will usually result in healthy LC's?


Quote:

with an LC it's a whole lot easier to tell that bacteria has won out because the LC goes clear.



English is not my first language, does "won out" mean the same thing as "won"? If so, then was that a typo or did you actually mean to say that when a LC finishes in a clear broth that it means that the LC is bacterial?


Quote:

Many people, for example, are spinning their LCs way too fast because they are using junk spinners from China, and those don't allow one to easily see when the mycelium wins out.



I have a hard time making sense of this as well. Regardless if you're swirling the LC by hand or by magnetic stirrer or don't swirl at all for that matter, how would you not be able to notice bacterial activity in the uncolonized parts of the broth? As long as the broth isn't yet 100% colonized by myc and there is a bacterial colony present, then surely the bacterial colony would be growing prolifically in the sterile broth not occupied by myc and thus the contaminated broth will become more and more turbid and therefore easy to spot.


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Re: Some notes about LCs [Re: Blue Helix]
    #26991988 - 10/18/20 08:29 PM (3 years, 3 months ago)

Quote:

Blue Helix said:
Quote:

mushhead said:
guys I need more notes.




You mean with reference to the dead spots in the trays?  I sometimes have that too, and I found it was because of the poor casing colonization (usually over colonized but under too).  I'd recommend that you work on your casing colonization.  It's too far colonized and bordering on overlaid, so the flush is sparse much like a tray without a casing.  With proper casing colonization, the flush will be about twice that dense.  Here is an example of a flush with proper casing colonization versus two without:

Ideal casing colonization:


Over casing colonization (and typical of no casing at all):


Under casing colonization:


All trays were LC trays, but in both cases of over and under casing colonization, you can see the reduced pin set.  The only problem with perfect casing colonization is that it is kind of hard for me to get consistently.  Even now I usually don't get it right, and I don't know why.  It seems to have something to do with the timeframe that the substrate transitions into rhizomorphic growth and could also be related to concurrent bacteria levels in the substrate. There might be a genetic component too.



:takingnotes:


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InvisibleD3_Myc
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Re: Some notes about LCs [Re: mushhead]
    #26992058 - 10/18/20 09:39 PM (3 years, 3 months ago)

I don’t have anything of substance to add other than my curiosity on LC and commenting so this continues to show
Up in my feed.


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Re: Some notes about LCs [Re: D3_Myc] * 1
    #26992093 - 10/18/20 10:15 PM (3 years, 3 months ago)

Quote:

D3monic said:
feed.



Yes, hungry lizards get fed


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Mateja]
    #26996384 - 10/21/20 01:15 PM (3 years, 3 months ago)

I wanted to update this thread with new information: I think the transferred tiny pieces to the LC were dying because of a bacterial problem, not because they were too small.  I made a mistake.  Sorry.


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OfflineCamera93
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Re: Some notes about LCs [Re: Blue Helix]
    #26996393 - 10/21/20 01:21 PM (3 years, 3 months ago)

:popcorn:


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Re: Some notes about LCs [Re: Camera93]
    #26996431 - 10/21/20 01:37 PM (3 years, 3 months ago)

Here we are, not quite 72 hours after inoculating this liquid culture into half gallon jars.  So much faster colonization than agar wedges.  This is why I use LC or at least LI.  With half gallon jars you need the speed.



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