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WBS failure (Trying again soon)

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H

hixidom

Rising Star
I'm going to try growing mushrooms again. This will be my second attempt. I want to start within the next few weeks, but I realized that I never really found out why my first attempt failed, so I hope that someone here can clarify my mistake for me so that I will not make it again.

I don't remember the tek I used. I boiled the bird seed to soften it, and then I sifted out the hard chaff and strained and rinsed the soft/plump seeds. Then I put the seeds in jars and pressure cooked for several hours. I let the jars cool and inoculated each jar before quickly putting it in an airtight trashbag (I did this in a bathroom that I had thoroughly cleaned beforehand). I put the trashbag and a thermometer on one side of a cabinet above my fridge, and I put a lamp on the other side. Over the next few hours, I moved the lamp closer to or further away from the bag until the temperature on the thermometer was correct and stable (I can't remember what temperature I used, but it was a supposedly optimal temperature that I had found in the tek I was using; like 80 degrees or something like that).

Anyways, days and then weeks and then months passed but nothing every grew in the jars. I thought that I would either get mushroom mycelium or mold, but I didn't think it was possible for nothing to grow at all. I never got a definitive answer on why this happened, but I always assumed that the WBS I used must have contained some sort of fungicide that I failed to wash off completely.

Has this happened to anyone else here before? Should I have boiled/rinsed the WBS more or should I have not used WBS in the first place? I think I will just use the standard vermiculite and brown rice flour next time, though I do have plenty of bird seed left.

Thanks for reading
 
If you put your jars in a sealed trash bag, there probably wasn't any gas exchange. 80 degrees is also too hot (the old optimal temp given was like 82F, but that was shown to be too high, iirc). As long as your house isn't cold, there's no need to heat the jars either. I've had jars grow just fine in 65F temperatures and other people have reported success in lower temps. The colonization time increases, but it also becomes harder for contaminants to grow.

I'd go with WBS over verm and brf...I've only used WBS to grow mushrooms and it has worked like a charm every time I've had it in proper conditions.
 
How long did you let the jars cool?

Was this a spore syringe or LC? Spores may be impeded by residual fungicide but live mycelium will not be.
 
SnozzleBerry said:
If you put your jars in a sealed trash bag, there probably wasn't any gas exchange...
Click to expand...

I don´t think this could have been the main reason for the failure. There are teks out there that advise growing mycelium in fully closed jars without any gas exchange mechanism, and it seems to work (I am actually in the middle of an experiment with this)

What I was thinking about was the light from the lamp. I think it is generally advised to grow mycelium in darkness (light being a bigger problem than slightly lower temperature). Was the bag really light-tight? (you can test it by puting it directly in front of the same lamp...)?

Also - are you sure about the content of your syringes? Do they come from a reliable source? (They Cubensis, right? :))
 
non-viable spores perhaps. It has happened to me before.

Even with no gas exchange you would get some growth in the beginning, I would think.

fungicides could be another problem.
 
Michal_R said:
What I was thinking about was the light from the lamp. I think it is generally advised to grow mycelium in darkness (light being a bigger problem than slightly lower temperature). Was the bag really light-tight? (you can test it by puting it directly in front of the same lamp...)?
Click to expand...
This shouldn't matter...I routinely colonize jars out in the open. Even light from a full spectrum CMH bulb does not impede colonization to any meaningful degree. This has also been observed by reliable individuals on several mycology boards.

EDIT: I just saw that the initial post stated that the grain was PC'd for several hours (does that mean 3+?). Depending on the type of pressure cooker used, pressure cooking jars for hours on end could have dried out the spawn.
 
I believe it is slightly harder for grain and seed to be colonized from spore. I would use a generous amount of liquid culture for inoculation (if you didn't already) no matter what you want to colonize. I've had so many simple grows fail from spores because they just would not take hold, but I've never had a single failure with liquid culture.
 
What type of seed was it? Was it a blend or straight millet.

When I've used birdseed in the past I try to get a pure millet blend as this seems to work the best. Mycelium doesn't seem to take to seed like sunflower seed very well; not sure why but I've noticed this to be the case.

I used a millet seed by Pennington bought from Wal-Mart for my spawn jars/bags and it worked like a charm.

Also, several hours in a pcooker seems like an AWFULLY long time for them to sterilize. I never cooked grain for more than an hour in the pcooker and never had a problem like you state here. Perhaps the grain was scorched or somehow the nutrients were broken down by the excessive heat?

Another reason could be, as somebody previously mentioned, that you didn't wait long enough for the grain to cool before innoculating. If this is the case the spores would have died from the heat and of course would not germinate.

More information could certainly help narrow down the cause of the problem. Hope this helps...
 
SnozzleBerry said:
...Even light from a full spectrum CMH bulb does not impede colonization to any meaningful degree...
Click to expand...

Thats interesting (most teks I´ve read advise darkness). Good to know - thanks for the info.

SnozzleBerry said:
...the initial post stated that the grain was PC'd for several hours ...
Click to expand...

Yes, several hours seems to be unnecessary. "My tek" says that 45 minutes is enough...
 
I think the reason you want darkness when growing myc is that light is one of the triggers for fruiting and you don't want your myc starting to fruit before it has full colonized the substrate. I could be mistaken.

grain teks usually involve 24 hour soaking to germinate all the endospores, cooking for an hour or so to moisturize and then PC for 45 minutes to sterilize.
 
How long did you wait for the jars?

Just last week, I found a jar I knocked up in January. It was in a drawer, and I had given up on it, but I left it alone. When I looked at it the other day, sure enough, mycelium!!

I'm pretty excited!

So, keep in mind it could take a looooong time.

From what I've read, a few things can cause slow germination. Mine was likely all three of the following:

Dry grain (the grain on mine turned out a bit drier than I wanted)

Older spores (mine were 2 years old)

Dry spores (mine were very dry, and I made the syringe not long before I shot up the jar)
 
Thank you very much for all the sound advice. To answer some questions, I was using a spore syringe, and the trash bag I used was black opaque. I don't remember checking its transparency, but I may have used 2 bags just to be sure that no light was getting it. It was a seed mix, but the sunflower seeds floated to the top when boiling/soaking, so I scooped them out. Otherwise, I don't remember much about this because it happened at least 2 years ago.

Something I just remembered:
After I gave up on the jars, I put them on a shelf in the garage. I planned on cleaning them out later. I never got around to it, and after a few weeks, the jars were filled with green mold. I guess something got in the jars from the air in the garage, but this might indicate that fungicide was not the problem. Maybe it really was bad spores, as Enoon suggested.

So far, I've learned:

1. 80+ degrees is too warm for mycelium growth.
2. More than an hour in the PC can dry out the substrate.
3. Pure millet is better than mixed seeds.
4. I need to learn what gas exchange is (I presume it means that the inoculated jars should be able to breath)
5. The jars should be completely cooled before inoculation.
6. Darkness may not be necessary for mycelium growth (I'll shoot for it just in case).

Thanks for all the advice. You have all been very helpful, and I'm sure I will succeed at this in the future (hopefully on my next attempt).
 
Orion said:
I would use a generous amount of liquid culture for inoculation (if you didn't already) no matter what you want to colonize. I've had so many simple grows fail from spores because they just would not take hold, but I've never had a single failure with liquid culture.
Click to expand...
seconded
 
I have tried to make LC's in the past and had no luck. I've only tried a few times, but after the first grow, spore prints were abundant and I felt like grain and multi-spore injections were easier.

Would anyone care to share a recipe they have used with success?
 
liquid cultures really are awesome. once you have LC you can innoculate grains and it speeds up the process a lot, plus success rates are very high. For LC make sure you use a sugar that does not caramelize. I.e. dextrose and malt work well; honey should work and works but not quite as good.

For gas exchange I always used several layers of protection - you want the jar to be able to breathe but you don't want dirt or bacterias etc. to get in. I used tyvek envelopes cut up to fit below the lid of my jars as the first and most fine-layer. In the lid there would be at least one hole filled with polyfill through which you can inject or use a self-healing injection port made of silicone (I've used bathroom silicone and it worked just fine for one round of PC, the second time I'd use it it would start to deteriorate, so renew every time you re-use the jar). Over the lid I put a coffee filter to keep out dust and stuff falling onto it. It's a bit overkill, but I'm rather safe than sorry with this kind of stuff.

For preparing grains this tek: Shroomery - From syringe to print using rye is really good. you can use it for WBS as well. There's also a lot of useful information here: http://fet.ezin.cz/files/roger_rabbit-mushroom_cultivation.pdf though it reads quite choppy. RR says tyvek should be on the outside of the metal lid (using two piece lids of mason jars) but I've had good results with one-piece lids and putting the tyvek on the inside and the polyfill etc. described above and never had a single contamination.

Hope you have fun with your grows!
 
That's good, there's a beer brewing store just down the street from me!

I think my problem is I have been trying with corn syrup. I think I'll also rig up my own stir plate, too.

Thanks for the good advice and links, Enoon!
 
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