Moss General Lab Notebook

hi all,

starting this as a general public log of my myco endeavors. nothing special at the moment, just some general notes.

04/04/2021
made transfers of shiitake strain, plates CA3 and CB3 ( ≈ P3 in Stamets’ system). agar wedge to honey liquid culture, 350 mL, jars CA3 and CB3 respectively. additional transfer of CB3 wedge direct to supplemented sawdust/wood dowels, jar CB3. sterilized 2 monotubs and transferred bulk pasteurized supplemented sawdust substrate from night prior to monotubs; transferred pearl oyster (CA3) colonized oat spawn to monotubs, 1 qt jar per.

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04/11/2021
transfer of shiitake CB3 liquid culture via 22G syringe to two supplemented sawdust/wood dowel half pint jars, 3 mL each. next transfer will be same process one week from now.

transferred additional 1 qt jar of pearl oyster CA3 colonized grain to 1 monotub. why? because it had to be used and that tub was lagging the other. will i introduce contamination pointlessly and spoil the batch? we shall see.

composted additional 1qt jar pearl CA3. overhydrated and half colonized, as is most of the batch due to reconstituting and re-sterilizing an old batch of unused grain jars gone dry.

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04/20/2021
transfer of shiitake CA3 liquid culture via 22G syringe to two more supplemented sawdust/wood dowel half pint jars, 6 mL each.

growth evident on two CB3 jars from liquid transfer on 04/11; growth from CB3 agar wedge ~3/4 through half pint jar from 04/04 transfer–slow growth but thick rhizomorphic threads.

spawn runs for pearl CA3 from 04/04 are likely a bust. contamination seems likely. better pasteurization methods might help but growth in the revived strain was sluggish from plates through grain. might just need a more vigorous strain. note–don’t be afraid to terminate at earlier stages in the future.

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Missed some updates! Reconstructing below:

05/10/2021
New culture started from spore syringe. Pleurotus Columbinus (Blue Oyster) transferred to 3 blank agar plates in SAB. Droplet transferred to flame-sterilized inoculation loop and spread in zig-zag pattern. Labelled C1.

Some Mysterious Day in Late May
Lentinula Edodes (Shiitake): Transfer of half of colonized wood dowels to old logs gathered last season. Do they have another fungus which has already visibly colonized? Yes. Are these the least visibly colonized of the logs. Yes. Do I dare to dream? Yes. Logs drilled and dowels hammered in. Plugs covered with candlewax. Set aside in the corner of garden storage space standing upright together. Let’s just see what happens in a year or whenever.

05/31/2021
Pleurotus Columbinus: Two C1 plates remained blank–no growth whatsoever. Third C1 plate fully colonized (and then a few extra days of my own delay). No contamination. Transfer of agar wedges from two most vigorous growth zones from third C1 plate to two new blank plates, labelled C2 in SAB. Flame-sterilized scalpel used for transfer; not a new blade. C1 plate placed in cold storage.

06/08/2021
Pleurotus Columbinus: C2 plates fully grown. No contamination. Transfer of scraped growth from most vigorous zone in each plate transferred to 1 new blank plate each in SAB, labeled C3. Flame-sterilized inoculation loop used for transfers. C2 plates placed in cold storage.

06/15/2021
General Lab Procedure: Two jars of honey liquid culture mixed. 350 mL, with addition of ~.025 g gypsum, ~2 g sawdust, glass shard in each. Sterilized in pressure cooker for a little less than 1.5 hours above 15 psi. (Previous sterilization over 1.5 hours lead to some carmelization). Moxed supplemented sawdust, 10:2:1/8::sawdust:wheat bran:gypsum. Can’t remember specific amounts mixed dried, but hydrated to feel and separated into four jars. Sterilized for ~3 hours above 15 psi.

Pleurotus Columbinus: Both C3 plates nearly fully grown. 1 plate presenting possible sectoring. Uniform growth plate chosen for transfer to liquid culture. Relabeled P1. Agar wedge transfer in SAB from P1 to two jars of honey liquid culture via flame sterilized scalpel (not a new blade). P1 and remaining C3 placed in cold storage.

Lentinula Edodes (Shiitake): 2nd batch of wood dowels transferred to 2 recently cut logs. Logs are clear of fungi growth, but came from a dead tree killed by insect infestation. Drilled and hammered in; plugs covered with candlewax. No major investment in this project. Doused in remaining liquid culture because: ¯_ (ツ)_/¯

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