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EntheoGod
Entheo



Registered: 03/06/15
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Last seen: 7 months, 27 days
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Is this normal for agar ? (New to agar)
#26799597 - 07/02/20 02:52 PM (3 years, 6 months ago) |
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I could spend a ton of time typing up a thread but the simple question is does this look normal for pasty plates or agar in general? Or is it bacteria? The last image is what it looks like before pressure cooking and the other two are after sitting a week after inoculating. Some look like this right after cooling inside the pressure cooker.

I’ll post more about the agar in this thread. I’ve been working to try finally getting clean spawn. I noticed none of the spores I’ve been using are growing at all on the agar even after about a week or a little longer. I know it can take weeks for germination to occur. Just wanna know if what I’m seeing here is expected and is something simple like the agar accumulated on the edges but it almost looks like what I’ve seen in pics of bacterial agar. There’s absolutely nothing showing on the surface of the agar just you can see this along the edges in nearly every agar plate I make.
I use the 123 agar recipe btw pressure cooking for 30 minutes after venting the pc for 10 before placing the weight on top and I allow them to completely cool in the pressure cooker before moving to an SAB. The black clump in the first plate is a group of spores that were shot inside it via MS Syringe.
I may have answered my own question by opening one of them and taking a picture. Don’t wanna show this because it’s embarrassing but here ya go. We live and learn!

Hmm gonna check a new batch without inoculating just to see if it’s related to the spores or my sterile tek.
Edited by EntheoGod (07/02/20 03:23 PM)
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Edmunter
Mr



Registered: 05/01/13
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Re: Is this normal for agar ? (New to agar) [Re: EntheoGod]
#26799648 - 07/02/20 03:17 PM (3 years, 6 months ago) |
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Come join us here.
https://www.shroomery.org/forums/showflat.php/Number/25662166
Your photos tell no story at all.
You usually have to wait at least 4 days till something happens
After a few days usually the condensation goes away but if it doesnt put a cup full of hot water on top and it clears it up
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EntheoGod
Entheo



Registered: 03/06/15
Posts: 648
Last seen: 7 months, 27 days
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Re: Is this normal for agar ? (New to agar) [Re: Edmunter]
#26799656 - 07/02/20 03:21 PM (3 years, 6 months ago) |
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Quote:
Edmunter said: Come join us here.
https://www.shroomery.org/forums/showflat.php/Number/25662166
Your photos tell no story at all.
You usually have to wait at least 4 days till something happens
After a few days usually the condensation goes away but if it doesnt put a cup full of hot water on top and it clears it up
Well these were inoculated around 7 days ago like I said. I figured it out. Definitely bacterial and will be scrapped. But I’ll be stopping in that thread for future troubles. Thanks for the response!
EDIT I originally said those were two days after inoculating but I misspoke I meant 7 days
Edited by EntheoGod (07/02/20 03:23 PM)
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Edmunter
Mr



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Re: Is this normal for agar ? (New to agar) [Re: EntheoGod]
#26799664 - 07/02/20 03:24 PM (3 years, 6 months ago) |
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Bacterial doesnt mean its toast. Let it grow out and see if you can see any decent growth. Then take that to a new tub, its the point of agar. In the meantime tart something new and get your sab work down.
Whats your understanding of how a SAB works?
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EntheoGod
Entheo



Registered: 03/06/15
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Last seen: 7 months, 27 days
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Re: Is this normal for agar ? (New to agar) [Re: Edmunter]
#26799699 - 07/02/20 03:39 PM (3 years, 6 months ago) |
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My understanding is an SAB Is a still air box. You want to keep air movement to a minimum inside it once you’ve sprayed the air with soapy water inside. The spores that are in the air will be dragged down to the floor of the SAB and they will stick to the walls as well. Over time that dries and allows the air to pick them back up again. The point is to move as precise as possible and never move your hand over the agar plate when preventable. Don’t touch the agar lid on the inside ever. I think that about sums it up am I right?
The thing with the agar you see above is it’s been about 8 days since inoculating and it’s not changed a single bit other than more of the yellow buildup around the edges and what you see in the last pic above. Is it normal to see a line almost like the surface of the agar appears different than the inside and bottom. I thought it was something to expect but got to wondering why I see no growth at all even when applying a pin to agar. Seems like something would have taken off. I decided to touch one of the plates a few days ago just to see what would grow and it started that night. Needless to say that agar was tossed anyway. I know that contams and bacteria germinate quicker than spores from cubes.
I’ll allow them to grow out more to see but I don’t expect to see any growth tbh I noticed the agar looking sweaty as well so I believe it would be best to scrap em eventually and start over but I would like to know what the error is in my process. From what I can tell it is related to using a spore syringe put to agar. Every spore syringe I’ve gotten recently from the same vendor has been not working at all or appears very weak on grains and won’t even germinate on agar.
Like I said though I’ll prepare a few more plates and leave a few Untampered with to see if something is getting contaminated in my process of inoculating or if it would have contaminated without me inoculating. I’ll also do a blank inoculation meaning I’ll take a transfer from an empty plate to another empty one to test if it’s maybe my actual process that’s causing it. Gotta narrow it down somehow!
Btw here’s my ghetto SAB
Edited by EntheoGod (07/02/20 03:44 PM)
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Edmunter
Mr



Registered: 05/01/13
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Re: Is this normal for agar ? (New to agar) [Re: EntheoGod]
#26799712 - 07/02/20 03:47 PM (3 years, 6 months ago) |
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Spores can take 4 weeks so as long as you flame slightly lift the lid and drop 1 drop in the middle it should tell you
OR make the tub with a hole tht you seal with heat resistant silicon and inject through that without opening the tub at all.
Loads of option bro. Where u from?
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EntheoGod
Entheo



Registered: 03/06/15
Posts: 648
Last seen: 7 months, 27 days
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Re: Is this normal for agar ? (New to agar) [Re: Edmunter]
#26799732 - 07/02/20 04:00 PM (3 years, 6 months ago) |
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I never have tried silicone injection ports but on agar with pasty plates I’m not sure that would be a great idea. I was thinking of inoculating through the GE hope in the lid of the plate and taping back over that with micropore tape but that feels unsafe to me. I never feel sterile opening the agar plates to inoculate them with anything. Most recent attempt started last night and I dropped some colonized grains on the fresh plates hoping to clean up some weaker growth that was on the grains before using them as spawn.
I’m from Kentucky the good ole Appalachian mountains haha
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