Not sure what should I call this project:
- TLV2023 MlmcMztpc
- IL2023-FNG mlmc mztpc
- IL23-FNG01 #mlmc #mztpc o #PC-MEL #PC-MAZ
Brief
First attempt at germinating the spores I brought with me. They are “non-conventional” spore prints, not done in a sterile environment.
- Mztpc: printed on drawing journal
- Mlmc: spores wiped up from print with masking tape…
Will attempt germinating them on a liquid culture and directly on grain (brown rice). I have done this before as a proof of concept. myo mztpc proof of concept
Also, don’t have proper instrumentation with me so will McGyver it. I have a jar marked at 1cup (250ml), ¼, ⅓, ½, and I know 1 teaspoon of sugar si approx 1.2g . Although am able to get alcohol, or alcohol wipes, I haven’t.
Also didn’t build my still air box yet, so… lots of “ifs” 😉
Glucose Solution 4%
1/2 cup water (125ml) + 1 tspn sugar (4.18g)
Grain: Brown Rice
Soaked overnight. Boiled 6-10 minutes
Get water to a boil then add the rice. Remove when rice still firm, not burst open. Air dry the rice and wait for it to cool down. Hummidity must remain within the grain. You don’t want (or as least as possible) condensation droplets within the jar. Bacteria like this starchy water conditions. Harder for them to grow on the “dry” surface of rice.
Mold will grow on the rice, no problem.
Jar Prep
Distributed the glucose solution into both jars. Distributed the rice into both (wide mouth) jars. Covered with aluminum foil caps.
You don’t need (nor want) it to be air tight. If air tight, it will create a vaccum when jars cool down, that will violently suction air from the surroundings when you open, pretty much contaminating everything.
Some people make this holes in the cap, and cover one with cotton and seal the other with high temp silicon. Helpfull if you will be doing this constantly, and only really worth it if everything else is just as sterile. Not really worth it if the spore prints are dirty. Chain will be as strong as the weakest link 😉
Sterilization
Default presure cooker cooks at about 110-120ºC
Mine is set 1.5 bar so (110ºC meh! Good enough)
Pressure-Temperature calculator:
Recommended to sterilize for 60-90 minutes.
Again, I didn’t have the time so 30 minutes. I’m banking on the cleanliness of the apartment to keep things clean.
Something I have done in the past to make sure everything is dead, is leave the prepared jars for a day at room temperature, before sterilizing. So any spores that might have contaminated my jars will be tempted to germinate.
LAZY PRO TIP
And then after hyphae/mycelium or bacteria (in non spore form) die off very easily. Spores are the tough ones.
Innoculation
Again, not ideal conditions, nor tools, but whatever. Life is tough
Basically a paper triangle snipped of the spore print into the liquid culture, and another into the grain jars, trying to get the spore side down in contact with the rice. I dipped the paper triangles in water before throwing in.
The spore-maskingtape strip was more of a pain in the ass, because it just wouldn’t let go of the cutter, so got my fingers involved. It’s fiiiine.
Next steps…
Covered everything up and put into zip-lock bags to limit/reduce evaporation and not have the rice dry up too fast. The weather here is very warm and dry.
Now we wait…
Reviewing photo log from the past. Last time I tried this drawing journal spore print was 2021. It took 10 days to get a visible mycelium growth. And 3-4 for contamination to show up jaja.
Also it was done on boiled roasted barley (with husk). I believe the husk might have helped fend of contamination. That inoculation was done in middle of the jungle with very precary conditions and so many spores and bacteria everywhere. Benefitfull bacteria I must add, just everything is SO alive there. You breathe and something grows.