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AlpineHighs
Thrill Shrieker

Registered: 04/22/20
Posts: 87
Loc: Montucky
Last seen: 1 year, 6 months
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Early Contamination? Again?!?!
#26853394 - 07/30/20 11:43 AM (3 years, 6 months ago) |
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Hey guys,
I inoculated a bunch of oat jars with agar wedges on 7/23, and it looks like my second attempt in two months is probably already contaminated. Or are they??
4 jars of B+ clone => agar ; 6 PE clone => agar ; 9 PE pin clone => agar
And every...single...jar looks like this after 7 days. I was worried about the weak/whipsy growth over the past couple days. This morning I smelled the GE hole of one with an SFD, and it smelled kinda similar to the 45 stank ass sour jars that I had to toss out a few weeks ago. So I took a couple away from my zone and opened them, and I'm 84.333% sure there was a distinct sour smell. There are patches of new rhizo growth on the bottom leading edge in every jar, but all jars have that weak/whispy growth below the grain surface, and fuzzy shit on top.
  
Oats were PCooked for 1hr 45min. Lids have correct size GE hole, with either an SFD or Bod's polyfil lids. I use a still air box. Proper sterile technique. All agar dropped to grain was on its 3rd transfer. All mycelium from each plate I used was this clean and rhizomorphic....
 
Is it worth shaking these jars to see if they recover? Or should I get them out of here ASAP, clean the fuck out of my apartment, and hope for better luck with round #3 of 2020?
I keep shaking and smelling the 2 I opened, and still can't definitively tell if they're sour. But the fuzzy stuff on top sure didn't look like mycelium when I opened them.
What the hell am I doing wrong???
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AlpineHighs
Thrill Shrieker

Registered: 04/22/20
Posts: 87
Loc: Montucky
Last seen: 1 year, 6 months
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Re: Early Contamination? Again?!?! [Re: AlpineHighs]
#26853580 - 07/30/20 01:26 PM (3 years, 6 months ago) |
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Anyone have any suggestions? I guess I'll plan to shake these jars and hope for the best.
I have these 3 PE T2 plates that I'm probably gunna use to inoculate more jars tomorrow. They look clean, but maybe dirty plates are the source of contams in my grain jars?
  
I know I should avoid the edges when cutting wedges from a full plate, but should I also avoid the center where the transfer wedge is? I have been using 1 plate (mostly full) cut into 4 equal segments for each jar. Is it bad practice to inoculate jars with the center bit that has the transfer wedge on it?
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jay.ach
Traveler



Registered: 05/25/19
Posts: 531
Loc: USDA Zone 8
Last seen: 18 days, 11 hours
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Re: Early Contamination? Again?!?! [Re: AlpineHighs]
#26854332 - 07/30/20 08:15 PM (3 years, 6 months ago) |
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-------------------- “One's destination is never a place, but a new way of seeing things.” - Henry Miller LAGM v. 2.023 LAGM2021
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AlpineHighs
Thrill Shrieker

Registered: 04/22/20
Posts: 87
Loc: Montucky
Last seen: 1 year, 6 months
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Re: Early Contamination? Again?!?! [Re: jay.ach]
#26855702 - 07/31/20 03:15 PM (3 years, 6 months ago) |
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Yeah I used Bod's oat prep tek, and even cut down on the boil time to make damn sure they weren't too saturated. As soon as I saw a few grains burst, I strained them and laid them out on a folding table to steam dry for almost 2 hrs. PC @ 15 PSI for 2 hrs.
I'm stumped as to which part of the process all this contamination is coming from. Lately I've lost well over 50 jars to sour rot; I think 4 or 5 different PC runs and SAB inoculation sessions.
I've been setting my SAB directly on a wood table after dousing every sq cm of surface area with 70% iso. Also using 1 or 2 blank petri dishes as the "floor" for all dishes I'm working with. I'll plan to use the lid of my SAB as the floor for this evening's inoculation sesh and see if that solves the issue?
Fingers crossed
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Fuddyduddy
Stranger

Registered: 04/29/20
Posts: 34
Last seen: 3 years, 1 month
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Re: Early Contamination? Again?!?! [Re: AlpineHighs]
#26855743 - 07/31/20 03:50 PM (3 years, 6 months ago) |
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Possibly underhydrated the oats..? If not fully hydrated & part of the PC cycle was used to finish cooking the oat to the center then perhaps not all of the natural bacteria was killed.
I cooked a batch just as you described and it was the only time I had bacteria in my jars. Cooking them until more oats burst has not lead to any problems for me. 40-45 min boiling
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AlpineHighs
Thrill Shrieker

Registered: 04/22/20
Posts: 87
Loc: Montucky
Last seen: 1 year, 6 months
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Re: Early Contamination? Again?!?! [Re: Fuddyduddy]
#26855755 - 07/31/20 03:59 PM (3 years, 6 months ago) |
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Damn. That makes sense. I've got jars in the PC right now...should I maybe let it run for an extra 20-30 min @ 15 psi, for a total of 2.5 hrs, to try and make sure bacteria is done for?
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