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DT2K
asdsadwvwv
Registered: 10/21/05
Posts: 69
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LC Expansion
#5818256 - 07/03/06 11:40 PM (17 years, 6 months ago) |
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Imagine I have a syringe of spores and a 500ml jar of malt. I inoculate the jar of malt and allow about a week and a bit for it to colonized thoroughly. Now imagine I take 30cc's of that LC and inoculate it into another 500ml jar of malt and wait for it to colonize in about a week as well. Now imaine I again take 30cc's of LC from the most recent LC and inoculate it into yet one more jar and wait for it to colonize. Graphically put: Syringe--->Malt--->Malt--->Malt
Is there any reason why my mycelium may be dead or somehow weak by the third jar? That is, does mycelium become weaker or can I technically do that for as long as I like?
Also, what happens to a LC if the mycelium eats up all or most of that nutrients in the malt (the malt becomes very light)? Does it die or does it just stop growing but lives?
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Omnicracker
Crusted Trolltivator

Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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Re: LC Expansion [Re: DT2K]
#5818296 - 07/03/06 11:51 PM (17 years, 6 months ago) |
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the best way to do this is to have a master jar, kept in a fridge. this is the first LC you make. if you need more, you pull a cc or so out and make another batch. when thats gone, start a new one with the master jar.
if you keep myc. in a constantly growing state, it will eventually show signs of senescence
Edited by Omnicracker (07/03/06 11:51 PM)
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DT2K
asdsadwvwv
Registered: 10/21/05
Posts: 69
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But is it likely that that would happen in 3 transfers as I have said in my example?
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Re: LC Expansion [Re: DT2K]
#5819272 - 07/04/06 09:45 AM (17 years, 6 months ago) |
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No.
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Blue Helix
bold hand


Registered: 02/02/03
Posts: 1,565
Last seen: 6 months, 18 days
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Re: LC Expansion [Re: DT2K]
#5821938 - 07/04/06 11:36 PM (17 years, 6 months ago) |
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When one expands mass mycelium out (i.e. LC or grain) on or both of two things usually happen:
1) Competing bacteria can catch up with the mycelium. This happens because bacteria always are faster than mycelium. In a healthy LC, there is always bacteria introduced from the spores or otherwise but overwhelming mycelium growth will stop the bacteria, push it into an endospore state. What happens when you expand out the LC? The weakened grasp of the mycelium network opens the playing field for bacterial growth, so initially both mycelium and the bacteria spring to life and grow. Given that the mycelium is still much more abundant, it will overtake the bacteria and the bacteria can go back into an endospore state again, but by the time the mycelium has captured the liquid culture the second time around, the bacteria will be more abundant relative to mycelium than it was the first time (because most bacteria grows a heck of a lot faster than mycelium). And what happens if you expand this new LC out again? Bacteria and mycelium spring to life yet again, but this time there is more bacteria relative to mycelium than ever. Can the mycelium overtake it yet another time? Maybe it can and maybe it cannot. Unless you are doing your work in a semi-conductor-level clean room and with state of the art equipment and had a perfectly pure monoculture of mycelium to begin with, at one level of expansion, be it the third or the thirtieth, the mycelium will lose vigor because it'll be spending more energy combating the ever increasing load of bacteria than growing. When dealing with LCs where you can't cut away from the bacteria like you can with agar (and you can't even see the bacteria), I try to keep my expansions down to less than 5 before going back to spores, using a piece of pure mycelium, or adding a tiny bit of hydrogen peroxide to the solution. If you keep trying to expand out without scouring the solution of bacteria in some way, you’ll likely run into a slowed growth from bacterial overload.
2) Another thing that _might_ happen after dozens if not hundreds of expansions and something that is often confused with number 1, is senescence or the natural aging of mycelium. Although this effect seems to vary vastly from species to species of fungi, it may happen to cubensis although it's never been studied in relation to cubensis to my knowledge. Keep in mind that some mycelium beds are the size of many hundreds of acres of land and they don’t seem to be suffering from senescence at all, so this effect might not even exist in cubensis. It’s just unknown at this time.
Edited by Blue Helix (07/04/06 11:42 PM)
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Omnicracker
Crusted Trolltivator

Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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i have experienced senescence with cubensis. i kept expanding grain to grain, but without keeping a masterjar. by the 5th generation, at least 50% of the fruits that came up were mutants, and the colonization/yield was slow and low.
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Shampioenier
Storm in aTeaCup


Registered: 07/29/05
Posts: 260
Loc: Milky Way Galaxy
Last seen: 17 years, 6 months
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it may have been that you had got your mushies into contact with cleaning chemicals of anti bacterial soap or any other chemical product maybe that caused the mutation and not, as you say, senescence. although its quite likely that u r right, i mean old retarded shrooms u know. like people. look pretty fucked up. even if their nice... mmm
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Shampioenier
Storm in aTeaCup


Registered: 07/29/05
Posts: 260
Loc: Milky Way Galaxy
Last seen: 17 years, 6 months
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i dont want to be rude but that picture with those two germans is revolting.
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