Thanks for the kind words guys!
Most likely, this is something everybody could figure out after months of searching forums and reading books. I did do the forums part, but not the books. Too expensive for this poor student. I combined techniques I found here, and on other fungi-related websites into one. Most likely somebody has done that before me, at least I hope so.
I know there are some.. differences between forums, their users and all that. I am not going to get involved by using names or locations.
Basically I used these modified statements from the people I see as the 'vets of fungi'.
- Cardboard is hard to contaminate
- Wood in agar culture is a great place for a culture to survive tough times in the fridge, even for cubes (so that means they do degrade/metabolize cellulose and/or pectin)
- Going from print -> syringe -> agar -> agar -> growing medium always leaves chance for contaminates; you'll have to expose your media/culture to air at least twice.
- Cardboard agar tek for woodlovers.
- Adding wood flakes to rye jars makes unusual growth happen.
Then, from my own knowledge as biotech student (which shouldn't add any value to my claims!):
- Yeasts and bacteria - from human source, the biggest contaminator around! - grow poorly on most cellulose based material, since they lack the enzymes to break it down. If they do grow on there, they would most likely reside on the inner structure than on the outside, since they have bigger survival rates surrounded by moisture and structures. When a colony gets big enough though.. Biofilms etc. etc.
- Cardboard contains nearly no sodium-salts or other nutrients that are favorable for bacteria. Just like with the husks of rye; the bacterial communities only secrete there, while they feed and thrive on the nutritious insides of the rye. Some contaminated jars didn't show contamination until I broke the cakes apart and found the rye to be eaten from the inside out; no fungal metabolites or signs of contaminations in some jars that were fully eaten by contaminants.
- Cardboard inoculated with agar wedges of cubes do great, are easy to work with - just tear a piece off, instead of cutting, leaving out one more contaminate source; the knife - and growth is vigorous when the cardboard is kept moist.
- Cardboard is easy to rehydrate; just add boiled toilet paper or boiled cardboard to the dish. And it's easy to see when it dries out; the edges turn a lighter color. This makes managing them easier. With agar, you'll just have to trust your gut, knowledge or experience to know when it's drying out (Sometimes agar doesn't shrink horizontally but only vertically, especially when using cling wrap).
- Cardboard is stackable and/or foldable which increases the effective surface of your petri dish with 200-300%; you can have more fungus in the same dish. Which translates into less space used, less materials needed and more jars to be inoculated with one dish.
- Cardboard can be of a lot of colors, adding better visibility to growth, status and furry/fuzzy things.
- Cardboard is CHEAP and a waste material, it can be found everywhere.
- Glue on cardboard is made with modified starch, which bacteria and yeasts CAN break down. Try to avoid glue. Also because it's treated with anti-fungal agents sometimes.
Some findings I thought I'd share:
Proven effective on:
- Trichorderma (which means this could still be a contaminate!), G. Applanatum, Reishi, B+, amazonian, Z-strain (B+ derivate?), Mexicana A, wine cap (strophoria?), pink oyster.
Oddities:
Cubes go very, very rhizomorphic on cardboard. Something that takes at least 3 transfers on agar for me.
Applanatum doesn't, but I guess that's it's thing: staying fuzzy.
Oyster goes the long route, leaving rhizomorphic growth out completely, and ending up stringy. I don't know how that's different on agar, since I haven't been able to get a uncontaminated oyster culture. It can also pin really fast from cardboard, so leave those in the dark or some shrooms might pop out of the petri dish.
Weird things observed when reversing this technique (placing cardboard on old - some contaminated - agar plates for the sake of experimenting):
- In Mexicana, the yeast on agar plates grows along with the fungus, this made me think that they were symbiotic. After some brain crushing, I figured out that Mexicana probably excretes more cellulase than it can take up sugars, thus leaving sugars for yeasts to feed on. Now I think the symbiotic relation is bullshit, why else would Mexicana secrete deadly metabolities when yeast infections occur? This information could prove useful in the need for biofuels; let mexicana break down wood, paper, plant materials, and let yeast make the oil/ethanol from the leftover sugars. Anyhow, mexicana seems to be able to break down either pectin, or cellulose but not both, since I can't dissolve plant tissue with metabolites or fungal secretions from mexicana. It does however weaken the cells structure. I don't own materials to further test this.
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- Seemingly dead agar plates (left untouched for.. ehr.. 6 months? at room temp. dried out, contaminated, pinned, total armageddon) seem to revive in 2-3 days after adding wet cardboard. Whether it's trich, contaminants or actually cubes is something I still need to find out. Not all species/strains of fungi do this of course, I'll report the findings when there is more to tell. I'm just doing this on the side when I have nothing better to do.
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- Sugars/nutrients in dried out LME agar are absorbed by the cardboard, making it a wonderful place for contaminates to grow, I haven't found a single one though.
Edited by JanSteen, 14 April 2016 - 03:17 PM.
