Mackload said:

hey, Bunny.12! welcome. you can do a few plates at a time, more the merrier. and if possible figure out how to take a picture of the surface through the lid. autofocus often settles on the lid...

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Bunny.12 said:

Mackload said:

hey, Bunny.12! welcome. you can do a few plates at a time, more the merrier. and if possible figure out how to take a picture of the surface through the lid. autofocus often settles on the lid...

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Hi Mackload, thank you for the welcome. I did do another plate but nothing seemed to show up yet. This is the best I can do with my camera as it is not a macro lens. I did take the picture in manual focus, cropped it and I got what I got. This is the third time I made up plates, the first time something grew, other than the one plate that had obvious contamination. I will try again tomorrow doing a few more plates. And then...  ...waiting again....

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that's the way. soon you'll have myc, and then you'll have mushrooms!

mushboy said:

Synack said:

Hard to imagine I'd get a vendor LC syringe of pure mold, no?

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its easy for me to imagine.

ive never done nats/orch or whatever but that just looks like straight mold to me. but i could be wrong.

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So I decided to go back to the original LC syringe that was kept sealed in my fridge, and start up two new plates with it. I do see what looks like a normal cloud of mycelium floating around in the syringe, and no other untoward characteristics, but I know that's hardly evidence of anything when it comes to LC. I'll come back in a couple days with fresh pix of what those plates look like and maybe they'll tell a different story.

Meanwhile, definitely don't take what I said as any kind of argument. If it's mold, it's mold. I have zero confidence in my own judgement here, which is why I asked in the first place. Trying my best to just do what folks here say. I just have this LC and no other way to make nats currently so I'm willing to beat my head against that wall a bit more before giving up.

I also held out hope that I could get my bacterial-as-fuck rusty whyte grain spawn (also from vendor LC) to top fruit, and I got one today! So, I submit as evidence of my reform and adoption of the Shroomery golden path for mush cult: three clone plates of tissue from that RW fruit. First time cloning, wish me luck!

Bunny.12 said:

This is the third time I made up plates, the first time something grew, other than the one plate that had obvious contamination. I will try again tomorrow doing a few more plates. And then...  ...waiting again....

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PuffM4gicDr4gon said:

How do you know you didn't spread mold spores all over your plate and healthy mycelium when you picked up the cut out piece? Spores are literally designed to be spread by air movement.

That's the main reason we cut out healthy mycelium, so it is removed from contamination, not the other way around.

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PuffM4gicDr4gon said:

If you're short on plates, look into reusable petri dishes or HG cups.

I have about 100 dishes that I just cycle through. I do no pours so I don't have to resterilize them before pouring. I prepare 30-50 at a time.

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PuffM4gicDr4gon said:

I don't often get mold or bacteria on my plates, but it does happen occasionally. I just make sure that I always have sterilized plates ready to go. If something looks fucky, I just cut out the healthiest bit and move on.

It only takes a couple weeks to grow out a plate to the size where it can be put to grain. Having peace of mind that I'm using 100% clean innoculant is worth the extra month to do a couple transfers. If you do a handful of transfers a week, you'll eventually get to the point where you always have a clean plate ready to inoculate.

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PuffM4gicDr4gon said:

I've seen enough circumstantial evidence to convince me that you are Doc Smoke.  I don't expect you to confirm or deny it, but either way, you being here is proof that you want to be a part of this community. So become a part of this community and listen to the people trying to help you. You'll fit in a lot better, have a less frustrating time, and get a fuck tonne of mushrooms in the process. This site has been here for 25+ years, there isn't really much that hasn't been tried yet. No one cuts contamination out of agar because it doesn't work. It's not some novel idea, just a bad one.

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Puffin said:

How many spores?

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A quick update on this one

About 32 hours after inoculation and there is some germination observed under the microscope



There is also an area with bacterial contamination



It's not a surprise that there is some bacterial contamination after putting so many spores onto agar from a spore print. If I had have waited another day I could have used antibiotic agar since those supplies just arrived yesterday. I'll store the plate vertically and see what happens.

This can be a good opportunity for experimenting with the antibiotics.



I'll document that here: Antibiotic Agar

PhenoDreamers said:

I've never had good success with antibiotics. Cabins and sandwiches are so much better

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anders said:

PuffM4gicDr4gon said:

Why did you cut pieces out of those plates that aren't MIB and Semp?

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I guess you are referring to me?  Some incel earlier said he is not looking at plates with "bites out of them".  Glad to not have his input.

Sometimes a plate in which you've invested four wedges develops trich or mold.  Rather than move four wedges, I cut out the bad.  Sometimes more comes so I cut that out.  I realize that this is not de rigueur but it is the way I do it, just like many things I do are not 'according to Hoyle'. (ancient reference)  Hell, putting a wedge into four quadrants is 'just not done'.  But I do it.

Back in the '70's Japanese cars were just crap and everyone knew it.  But the makers zeroed in on the actual problems with clarity and focus, and fixed them one-by-one, and I'll bet you drive a Japanese car now.  It's called the Scientific Process.

I was aware of trich and how it looks, but was fooled by mould.  Now I am aware of that and am mitigating.  Sometimes the only option is to remove the last good wedge and put it on a fresh plate, which is what I've done with MiB.  Today I did a mass innoc to grain, and I know that at least 4 jars are going to be pure mould.  But I will let them grow because I want to see what that actually looks like.  Of my 14 jars (all catalogued) I hope to have at least some rip nicely.  But time will tell.  Have I introduced dlean myc?  Have I hydrated the grain to the correct level, etc?

What I know though is the UV sterilization flood is a good decision, as were several others I've learned.  Bit-by-bit.

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PhenoDreamers said:

Yeah they didn't do shit for me dude. Gentamicin

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Puffin said:

You added the antibiotics to the agar after pressure cooking?

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RoscoeReturns said:

Puffin said:

You added the antibiotics to the agar after pressure cooking?

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Gentamicin is heat stable and will survive autoclaving just fine. You can add that to your broth before you sterilize.

Chloramphenicol is NOT heat stable. After you autoclave your media you must wait until it cools before adding chloramphenicol. It must be added aseptically to your liquid media just before you pour. Chloramphenicol is also degraded by hydrolysis in an aqueous vehicle. Use those plates soon after you pour them, they don’t retain activity for very long. Refrigeration can slow that process.

Malt extract agar already inhibits the growth of most bacterial species due to its acidic pH. Key words being inhibits growth, not kills off. I’m currently experimenting with using tryptic soy agar media to see if this growth inhibition is causing “false negatives.” If bacteria is present yet not able to grow am I seeing a clean plate that is not really clean?

Having said all that, I have used antibiotic agar in the past to clean up very dirty cultures that I couldn’t otherwise replace. Other than that very rare (for me) instance, I don’t see the utility of it in cultivation. I’ve had equal success using cabin sequestering, so I don’t know that I’ll go through the added steps of making antibiotic agar again.

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anders said:

Bunny.12 said:

This is the third time I made up plates, the first time something grew, other than the one plate that had obvious contamination. I will try again tomorrow doing a few more plates. And then...  ...waiting again....

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If you're using syringes (not reommended) rather than spore prints you're much more likely to get contamination.  Any little bit has all it needs to multiply in that syringe.  Whereas you take a small sample from a spore print, reducing the chance of contamination.

In either case it's a good idea to use a cell spreader to spread out your spores so they can germinate with some room around them and you can see what you have.

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Hi, thank you for the information. I just bought the syringe of spores as it was one of the few places selling material in my country. I had no luck germinating spores on previous plates so I just left the drop of solution in the corner hoping two spores meet up. Just for kicks I will be trying to isolate a few of these sites and transfer them onto new plates. Of course I can not find any stainless steel wire (it is in a box here somewhere) but then I had a thought. I could use the end of a guitar string as one.

ThurmanMurman said:

My brother in Christ...

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