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 Dr_Smegma
impersonates a guy who's a TV Dr
Registered: 01/15/18
Posts: 87
Last seen: 2 years, 2 months
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 Help figure out where contaminant came from 
#25162772 - 04/24/18 04:41 PM (5 years, 11 months ago) |
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Hey guys I'm pretty sure I know the source of the contaminant but I'm hoping you guys can either agree or help point me in the right direction.
I inoculated 7 grain jars via agar with GT clone from a second flush. I shook each grain jar and then opened and smelled each one of the jars before hydrating for 10-15 minutes or so with tap water. They all smelled like fresh mushrooms before spawning to the mono tub.
During my pasteurization of the substrate the temps reached 170F for 7 or 8 minutes before starting to drop. This is where I think I ran into a problem. My guess is I partially sterilized the substrate and made it easy for bacteria in the air or on myself to grab hold of the substrate before the mycelium. There was absolutely no growth of mycelium on any part of the substrate (from what I could see from the top) after 6 days but a horrible rotten garbage smell emitting from the tub.
Does this seem logical that I just partially sterilized the substrate and thus during the open air spawn to a tub bacteria got ahold of the substrate first? Other than being more careful with pasteurization temps in the future is there anything I can do to combat bacterial contams?
After emptying the contamed substrate into a garbage bag (x3) I soaked the tub in bleach water for an hour, then washed out with a soapy rag, then dried it partially to give it a final rub down with 70% iso. Is this tub good to go again? There was a liner around the substrate.
Below is a copy of my offline grow log so you can see all my steps for yourself if that helps, no pics, sorry:
Grow Log GT Clone from 2nd flush 3/28
Spawn: Yes - Rye Berries
Substrate: Coir, Hpoo, Gypsum, Verm, Water - Amounts below
Inoculated via: Agar Wedge
?/?
Cloned biggest fruit of second flush from GT cake - 1.8g dried.
?/?
Transferred from clone agar to other agar dishes to ensure clean growth - 1 dish contained 3 look amazing.
?/?
Tested dried fruit by itself - level 2 trip on 1.8 grams, 1/8th should be a fun time
3/28
Prepped 7 quart grain jars in PC 2 hours at 15 PSI.
3/29
Did Agar to Grain transfer in still airbox. Cloned GT fruit to 7 quarts rye berries.
4/17
Prepped bulk substrate as follows:
1/3 brick of coir + 7/8 quarts water
1&1/2 cups gypsum
1&1/2 quarts Verm
1.5lb hpoo
~ 2 quarts H20 (1.5 quarts first then another .5 added to field capacity)
5 quart jars full
Pasteurized between 140F and 170F for 1 hour — Temp hit 170F for 7 minutes
4/18
Shook grains and smelled - Soaked grains in jars for ~15 minutes
Spawned 7 jars (~ 5+ quarts) of grains to 5 quarts of substrate
4/23
No signs of any growth on substrate  WTF happened?
Strong garbage smell noticed coming from tub, removed from grow room and put in garbage bag for the night until I can discard
4/24
Disposed of substrate, cleaned tub - bleach water soak, rubbed down with soapy cloth, rinsed, rubbed down with 70% ISO
Edited by Dr_Smegma (04/24/18 04:44 PM)
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P.Avgvst
Registered: 03/13/16
Posts: 56
Last seen: 4 months, 24 days
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Re: Help figure out where contaminant came from [Re: Dr_Smegma]
#25163017 - 04/24/18 06:31 PM (5 years, 11 months ago) |
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Yeah, sounds like a badly done pasteurization. Try keeping the temperature high for longer, and to not let water accumulate in the tub, or it will rot.
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Enkidu
"No-Such-Person"
Registered: 07/09/16
Posts: 10,698
Last seen: 8 months, 27 days
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Re: Help figure out where contaminant came from [Re: Dr_Smegma]
#25163029 - 04/24/18 06:35 PM (5 years, 11 months ago) |
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--------------------
Within You , Without You
      
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Dr_Smegma
impersonates a guy who's a TV Dr
Registered: 01/15/18
Posts: 87
Last seen: 2 years, 2 months
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Re: Help figure out where contaminant came from [Re: P.Avgvst]
#25163031 - 04/24/18 06:36 PM (5 years, 11 months ago) |
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Quote:
Yeah, sounds like a badly done pasteurization. Try keeping the temperature high for longer, and to not let water accumulate in the tub, or it will rot.
I thought the idea was to pasteurize your substrate in the 140F - 160F range for an hour. Wont higher temps for longer times partially sterilize the substrate letting bad bacteria and other contams get a foot hold on the substrate before the mycelium?
I was guessing I partially sterilize the substrate from just being at or over 170F or 7 minutes.
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Enkidu
"No-Such-Person"
Registered: 07/09/16
Posts: 10,698
Last seen: 8 months, 27 days
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Re: Help figure out where contaminant came from [Re: Dr_Smegma]
#25163049 - 04/24/18 06:42 PM (5 years, 11 months ago) |
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Read the link I posted
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Within You , Without You
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Dr_Smegma
impersonates a guy who's a TV Dr
Registered: 01/15/18
Posts: 87
Last seen: 2 years, 2 months
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Re: Help figure out where contaminant came from [Re: Enkidu]
#25163072 - 04/24/18 06:51 PM (5 years, 11 months ago) |
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Quote:
Maybe it was the soak of grain jars in water before spawning but idk
https://www.shroomery.org/forums/showflat.php/Number/24077162/fpart/all/vc/1
I was thinking the same thing. It had to either be the slightly high temps during pasteurization or the grain soak after they were fully colonized. Since those were the only things I did differently from the Mono tub that was started 2 days prior to this one that looks beautiful.
But when I searched I couldn't find anything about the grain soak causing contams for anyone so I was doubtful.
I'm using Hpoo so i'm not sure the method of pasteurization you linked will work for me, I've been using this one: https://www.shroomery.org/forums/showflat.php/Number/17246844#17246844
From what I've gathered, since I am using Horse poo, any partial sterilization (more than a minute or two above 170F) will be just as bad as having an unpasteurized substrate.
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P.Avgvst
Registered: 03/13/16
Posts: 56
Last seen: 4 months, 24 days
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Re: Help figure out where contaminant came from [Re: Dr_Smegma]
#25163074 - 04/24/18 06:52 PM (5 years, 11 months ago) |
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Quote:
Dr_Smegma said:
Quote:
Yeah, sounds like a badly done pasteurization. Try keeping the temperature high for longer, and to not let water accumulate in the tub, or it will rot.
I thought the idea was to pasteurize your substrate in the 140F - 160F range for an hour. Wont higher temps for longer times partially sterilize the substrate letting bad bacteria and other contams get a foot hold on the substrate before the mycelium?
I was guessing I partially sterilize the substrate from just being at or over 170F or 7 minutes.
No, man. Sterilizing is way harder. You should go to around 170 F for a good pasteurization and to kill most of the fungus cells, yeast and most mesophilic bacteria. Less and you kill them only partially, and in a nutritious media as a bunch of cooked rye, they’ll be multiplying in minutes.
The link Enkidu posted is REALLY good.
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Dr_Smegma
impersonates a guy who's a TV Dr
Registered: 01/15/18
Posts: 87
Last seen: 2 years, 2 months
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Re: Help figure out where contaminant came from [Re: P.Avgvst]
#25163080 - 04/24/18 06:55 PM (5 years, 11 months ago) |
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Cool so when I pasteurize my coir/hpoo/verm/gypsum substrate I want to make sure I'm getting the temps up to 170F-ish for a longer amount of the time?
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Enkidu
"No-Such-Person"
Registered: 07/09/16
Posts: 10,698
Last seen: 8 months, 27 days
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Re: Help figure out where contaminant came from [Re: P.Avgvst]
#25163082 - 04/24/18 06:56 PM (5 years, 11 months ago) |
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Oh my bad I didn't realize you used poo know nothing about poo
--------------------
Within You , Without You
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Dr_Smegma
impersonates a guy who's a TV Dr
Registered: 01/15/18
Posts: 87
Last seen: 2 years, 2 months
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Re: Help figure out where contaminant came from [Re: Enkidu]
#25163299 - 04/24/18 08:41 PM (5 years, 11 months ago) |
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Quote:
Oh my bad I didn't realize you used poo know nothing about poo
From what I have read Coir + verm substrates you can partially sterilize because they're extremely resistant to contams. With Hpoo you only want to pasteurize it because the partially sterilized substrate with poo in it is super easy for contams to grow on.
Hopefully someone with more knowledge of working with poo will chime in on this. I might also just try a 50/50 substrate of coir and verm too.
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Enkidu
"No-Such-Person"
Registered: 07/09/16
Posts: 10,698
Last seen: 8 months, 27 days
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Re: Help figure out where contaminant came from [Re: Dr_Smegma]
#25163322 - 04/24/18 08:49 PM (5 years, 11 months ago) |
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Yeah I would wait for someone with knowledge on poo
Find a poo tek and it should say but sounds like you already did.
If that wasn't it I would guess maybe the water you used unless you boiled it or something before using to hydrate,maybe that introduced some contams or fucked with the grain ? But idk much about hydrating grains before spawning either, wish I could help more
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Within You , Without You
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