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Offlinepjeterschorstein
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Bacillus / yeasts?
    #25351894 - 07/28/18 10:59 AM (2 years, 9 months ago)

Hey all,

I made 33 jars agar to rye. unfortunately, after 10 days half of them showing slimy signs and it's smelly which I guess is Bacillus or yeasts.
http://

Some I made from loop inoculation to agar to grain and some MS to agar to grain and I didn'T see anything unusual on the agar.
I PCed the jars for 90min and inoculated them in a glovebox. I had this contam just once before and I am not sure what happened.
One thing is that I used more agar in an attempt to make it faster and maybe the extra  moisture through it off.. Second is that I used smaller holes on this jars (4mm) maybe there was not enough gas exchange? another thing is that they were placed in a room without much ventilation ( I moved the spawn to my fruiting area now).. Also, I used less gypsum.
Still, I'm not sure how the spores got into the jars.
can I use the remaining jars as masters or is there a chance they'll cary it also?


Edited by pjeterschorstein (07/28/18 01:02 PM)


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Re: Bacillus / yeasts? [Re: pjeterschorstein]
    #25352093 - 07/28/18 01:20 PM (2 years, 9 months ago)

They obviously got in during inoculation.


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Offlinepjeterschorstein
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Re: Bacillus / yeasts? [Re: bodhisatta]
    #25400113 - 08/20/18 09:33 AM (2 years, 8 months ago)

Hey Bod can you expand on that? you sure it's not from the grains from before?
I tried again. Seems like I'm gonna have some problem again (not as much but still)
I use a mask, closed room. I put gloves, sanitize them, , spray sanitizer in the air, open pc, then wipe the jars, also I clean the grain master or agar plates before putting them in glovebox .
Yesterday I scrubed the glovebox with soap , then with alcohol because I thought it might be the source.
It's frustrating. Just had to deal with trich last two months, managed to get rid of it and now this..


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Re: Bacillus / yeasts? [Re: pjeterschorstein]
    #25400196 - 08/20/18 10:30 AM (2 years, 8 months ago)

There's contaminants in your jar that would have died even if you did a half assed sterilization


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Re: Bacillus / yeasts? [Re: bodhisatta]
    #25402288 - 08/21/18 08:27 AM (2 years, 8 months ago)

Was just reading some post you wrote about glove box (which I've been using recently) vs SAB. Feeling like a moron now how I've went through half of this site posts already and this basic thing escaped me..
Also, Ive been using much less gypsum lately so maybe the acidity contributed to how easy the contam took over. would try to up it now.
Hope this solves this annoying episode :thumbup:


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Re: Bacillus / yeasts? [Re: bodhisatta]
    #25432776 - 09/03/18 01:17 PM (2 years, 7 months ago)

Well, now I'm really not sure. I got tyvak shirt and hoody, sprayed the air with lots of lysol. cleaned everything like in RR video. close room, STB. showered and brushed my teeth before + trying not to breath heavily. I inoculated 8 rye spawn jars grain to grain on the 23/8, shook it at 40% on the 30/8 . now 2 jars showing signs of wetspot.
Can it be from the grain master somehow or maybe this rye I bought is no good?
I can see they'll turn bacterial because some of the grains are shiny looking before they make white ring.
Please help.. I din't have this problem before and I'm loosing too much spawn already...
Also, I think it got to my oysters because I'm smelling two buckets that are slow to colonize and I think they don't smell right..



Edited by pjeterschorstein (09/03/18 02:42 PM)


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Re: Bacillus / yeasts? [Re: pjeterschorstein]
    #25433104 - 09/03/18 03:52 PM (2 years, 7 months ago)

Spawn bad? Grains in recieving jars not prepped properly? Bad sterile technique during transfer?

I'd mentally go through each step and find where you can improve.


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Offlinepjeterschorstein
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Re: Bacillus / yeasts? [Re: Wing]
    #25434272 - 09/04/18 04:25 AM (2 years, 7 months ago)

Wing, I am over it mentally all day and made changes as I wrote above but still it happens.

My question is if it could be from the spawn master I used on all of them?

Bodhisatta said it must be from inoculation.. but I can't see how (maybe a few contams but not 50%+..)
I prepare the grains just like in RR video and inoculate them also accordingly.
btw. I use the same jars but soak them with plumbing powder 12h then clean with soap and bleach.

:sad:

Think the only upside is if I learn something from that.


Edited by pjeterschorstein (09/04/18 04:30 AM)


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Re: Bacillus / yeasts? [Re: pjeterschorstein]
    #25434778 - 09/04/18 12:57 PM (2 years, 7 months ago)

It would seem your master jars are contaminated. Do you have pics of the agar plates before you put it to grain?


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Re: Bacillus / yeasts? [Re: Wing]
    #25434976 - 09/04/18 02:22 PM (2 years, 7 months ago)

Wing, no I don't. It was a few different plates: clone, MS and loop inoculation but it all comes from 13/7 and before this I never had wetspot. I know I used a lot of agar that time and it made it worse.
I think I'll start over with agar now, make new master spawn and get rid of all the agar from before. Also, I think whatever spawn survive from before, I'll give it a shake to see if it recover before I spawn it (if it's survive it's good, right??


How do you spot wetspot on agar anyway?


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Re: Bacillus / yeasts? [Re: pjeterschorstein]
    #25435030 - 09/04/18 02:49 PM (2 years, 7 months ago)

Wetspot is such a shit generic term. You spot contamination. Usually its obvious sometimes its a little harder. You post your agar and get critique that's how you learn more


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Re: Bacillus / yeasts? [Re: bodhisatta]
    #25435079 - 09/04/18 03:09 PM (2 years, 7 months ago)

Ok. Thanks. It's hard to see but that's what I thought of tossing (2 different strains, all MS) I made some more I can post in a few days.


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Re: Bacillus / yeasts? [Re: pjeterschorstein]
    #25435206 - 09/04/18 04:34 PM (2 years, 7 months ago)

Cant tell much but i personally wouldn't use the pictured dishes. Try to do transfers before the mycelium consumes the entire plate


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Re: Bacillus / yeasts? [Re: bodhisatta]
    #25436924 - 09/05/18 09:50 AM (2 years, 7 months ago)

Yes, I thought so too. I threw them anyway. I made these on the 29/8 and I made a few more some days after. One of these had trich. That's first time I had trich on agar. It happened also recently on a plate with wild mushroom clone. both happened after I switched to SAB from GB but I think it's because I wiped it down instead of leaving it wet.
Sorry for the long post I really appreciate your help :sun:


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