Home | Community | Message Board


This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: North Spore Injection Grain Bag   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Left Coast Kratom Buy Kratom Extract   Kraken Kratom Red Vein Kratom   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Buy Bali Kratom Powder   Mushroom-Hut Substrate Bags

Jump to first unread post Pages: 1
InvisibleTFI
Stranger
 User Gallery

Registered: 04/06/14
Posts: 344
What’s wrong with my agar
    #28319362 - 05/14/23 10:13 AM (8 months, 10 days ago)

Hello all back again to seek some help.

Backstory: a year ago had some incredible Cambodian genetics rocking along but had to destroy the process due to moving, I let a friend save some wbs jars for me until I could start again. I took the wbs and put it to agar and let that colonize out and restarted. I was not paying attention to every detail and now my current jars have a slight bacterial/mold contamination. I didn’t vent my pc long enough and had bad sterile technique in my SAB. Still got some fruits but far from what it needs to be.

Soooo recently I took a wbs master jar and a clone from one of the fruits and put that to agar hoping to save what I have going. I was reading a lot and discovered that bacteria can embed itself into the mycelium and it’s hard to tell if it is clean or not.

I have never had this many problems with contamination and I want to order some more spores to completely restart everything but in the meantime I wanna see what’s happening with my agar as of now.

I’m using 5gs of agar agar powder and 5gs of light malt extract and 250ml of water. I vented the pc for 10 min and then in my SAB I wipe down with alcohol and spray everything with soapy water and wait for 5 min before doing my work. Flame sterilize the tools and do my best to be quick and clean here’s the best pictures I can get so far

This is from the clone, you can see tight growth in the center then it gets very very light and idk what that is.



Clone transfer#2 from same original clone jar (notice the bacteria)




This is from the wbs master jar transfers.

Transfer#1



Transfer #2


As you can see it looks much thicker and healthier to me.

Sorry for not having better pics.


Extras: Filter Print Post Top
OfflineB Traven
Stranger
Male User Gallery


Registered: 03/10/20
Posts: 2,478
Loc: Central Megalopolis
Last seen: 1 hour, 55 seconds
Re: What’s wrong with my agar [Re: TFI] * 1
    #28319403 - 05/14/23 10:53 AM (8 months, 10 days ago)

I've got a few cultures like this. With the sort of scenario you describe, I think it's pretty easy for bacteria to sneak into a culture in the grain shuffle and then hitch a ride right back onto the agar.

Lately I've been trying to hold onto clone lines that have gone onto grain and proven themselves, basically just keeping the flame alive by going back and forth between agar and grain. I treat each G2A plate as genetics that I like, but presumably in a dirty culture. I really look hard for clean growth before deciding that I'm ready to go back to grain. There's lots of things I want to try in this process, but haven't gotten around to yet. I'm saving most of them for the most recalcitrant lines, the ones that I can't seem to clean up with just sequential transfers.

One thing I've learned is that some of these cultures really don't "look" off. It's only by noticing the unusually high contamination rates and general fussiness in those lines that I detect the problem.

I think the most important thing to try to do whenever possible is take growth from the leading edge of the mycelium.

Another thing I think a lot about is where we are in the materials chain. One of the issues with sequential grain transfers is that the old grains are still hanging around. Any bacteria in the culture are going to have an easier time multiplying on a four-month-old grain with senesced myc that they've been hanging out on the whole time. I think the best-case scenario for a really clean culture on really clean grain is that you end up with a few old, dried, shrivelled-up grains in the mix. This, in my view, is the main reason to go back to agar. You're separating the living culture from old materials. So, let's say you drop a grain onto a plate. Let's assume the grain is just covered in bacteria. The myc hits the new plate and tries to outrun it. You cut a bit from the leading edge and transfer it. That new wedge is comprised of 100% new material, but it was in the same room as that nasty bacterial grain. It got a bit of bacteria just from being in that environment, the same way one might smell like cigar smoke after hanging out with their drunk uncle for a few hours. So run another transfer. That transfer was only in the same room as the wedge that came from the grain plate, so its overall exposure was less. But maybe still significant. Do that a few more times, and, from my point of view, you can breathe a little easier.

If you let a plate get completely colonized before transferring, and can't take transfers from the leading edge, and there are still bacteria hanging around, then they get a chance to catch up. And that whole process is negated, at least to some extent.

Other things I have yet to try but that you should definitely read up on are water agar, the sandwich method, hot pours, and the JoseX poke. I've got a few cultures that are definitely getting the JoseX poke treatment if they give me any more trouble.

Also, remember that no spore print is 100% sterile. If you grow out plate pins and take swabs from them in a sterile environment, the resulting spores aren't likely to be any dirtier than what you'd be ordering. Even if your spawn is bacterial as fuck.


--------------------
Beware of advice- even this.


Edited by B Traven (05/14/23 10:56 AM)


Extras: Filter Print Post Top
InvisibleTFI
Stranger
 User Gallery

Registered: 04/06/14
Posts: 344
Re: What’s wrong with my agar [Re: B Traven]
    #28319483 - 05/14/23 12:08 PM (8 months, 10 days ago)

Ahhhhh very interesting. Makes since as to why everyone says take the leading edge. In my little pea brain I assumed take a sample from just the inside of the leading edge because it’s already established and growing stronger than the leading edge.

Dumb lol

It’s nice to get information on other techniques too like the sandwich. Have never heard anything about the joseX poke either so I’m about to do some research.

How many transfers would one recommend doing? I’m sure it depends on what you’re working with and how you work but is there a general rule? I usually do 3-4 transfers before adding agar to grains. Perhaps I should do 5-6.


Extras: Filter Print Post Top
OfflineB Traven
Stranger
Male User Gallery


Registered: 03/10/20
Posts: 2,478
Loc: Central Megalopolis
Last seen: 1 hour, 55 seconds
Re: What’s wrong with my agar [Re: TFI]
    #28319512 - 05/14/23 12:36 PM (8 months, 10 days ago)

Yeah, my minimum is 3-4 times, but I often feel more comfortable with 5-6. It's also a chance to expand out to more plates and just observe the growth a bit more. Sometimes I'll intentionally take multiple transfers from one plate, but then on the next round only select a couple of the best-looking new plates and discard the rest.

I used to let my plates completely colonize and then take transfers from them. Couldn't figure out why I never outran anything lol


--------------------
Beware of advice- even this.


Extras: Filter Print Post Top
InvisibleTFI
Stranger
 User Gallery

Registered: 04/06/14
Posts: 344
Re: What’s wrong with my agar [Re: B Traven]
    #28319610 - 05/14/23 02:33 PM (8 months, 10 days ago)

Hahaha sounds like me. I have probably never done it right to be honest. I just see what I think is healthy growth and go from there. At times I think I have it figured out and then boom reality hits like a ton of bricks.

I have a hard time throwing things away at times. I will often let the agar go till it pins especially on plates I have already taken from.

I love being able to talk to people about it too, I have learned so much. So many little nuances that can easily be overlooked, idk how many times I get depressed thinking I have lost my touch only to realize that I made simple little mistakes in the beginning.
Like having what I think are quality colonized jars only to find out they were contaminated. The smell test is ok but it’s far from perfect.


Extras: Filter Print Post Top
Jump to top Pages: 1

Shop: North Spore Injection Grain Bag   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Left Coast Kratom Buy Kratom Extract   Kraken Kratom Red Vein Kratom   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Buy Bali Kratom Powder   Mushroom-Hut Substrate Bags


Similar ThreadsPosterViewsRepliesLast post
* Re: mold in agar plate Anonymous 12,180 3 12/04/99 12:35 AM
by Anonymous
* Fruit on Agar?? Yarry 5,049 17 01/22/04 09:44 PM
by Anonymous
* Agar to Rice flour substrates foxxybrown 2,136 3 09/01/03 05:00 AM
by eggy
* Agar Anonymous 1,538 8 04/15/04 10:15 AM
by Bi0TeK
* golden teacher agar asking fugu 3,052 6 01/31/03 12:59 PM
by debianlinux
* agar/peroxide conc. iudexk 1,325 1 08/27/01 01:17 PM
by jonnyshaggs420
* Why grow on agar? stcore 2,012 1 07/29/02 05:03 PM
by bluhoney
* no signs of colonization after 3.5 days - what went wrong?
( 1 2 all )
laughingbuddha 2,389 20 12/04/04 11:10 PM
by agar

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, LogicaL Chaos, 13shrooms, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a
269 topic views. 17 members, 188 guests and 63 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.018 seconds spending 0.005 seconds on 12 queries.