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BExa
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Two years now, unable to get clean spawn. 100+ jars bacterial.
#28441209 - 08/21/23 01:03 AM (1 year, 4 months ago) |
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Been in the hobby for about two years at this point and every single spawn jar I've ever made has turned out bacterial(probably 100 jars at this point), or at least that's what I believe the contam to be. All jars fully/nearly fully colonize but suddenly begin to display signs of bacterial growth(condensation, wet looking uncolonized grain pressed against the glass, smells sour on spawn to bulk). I have been using BODS Monotub/Oat tek and jars with synthetic filter paper stickers from Amazon. For the first few, the contaminated jars would still fruit when spawned. I'd get about 7 oz on the first flush then maybe an oz or two more, then the sub would seem to become overwhelmed by the contam and I'd toss the substrate(the sub would develop the same sour smell as the jars. When first colonizing the coir sub, the tubs smell fine. Its if the myc out runs the contam but the contam eventually catches up.) Most jars have been oats of which I've tried two different suppliers now. Triple crown and Atwoods brand. I have also recently switched to white millet from nrsworld.com as I suspected my grain to be the issue. I used Cracks no prep millet tek. Yet again, the first run with millet has the same exact issue. I've used multiple different cultures. I started out from spores, grown out on agar. I probably grew out 100 or so plates. Id say probably 90-95% of the plates I poured turned out completely clean. No growth unless inoculated. So I believe my sterile technique to be on point. My two most recent runs were from liquid cultures supplied from Myyco. One syringe was use on oats, the other was millet. Same issue. At this point I am completely dumbfounded and not sure what is going wrong. I've changed cultures, changed grains. I've tried SAB and fans with merv 13 filters. Poured plenty of clean plates with both but ended up going back to the SAB the latest two rounds as I got rid of my fans. The only thing I can think is either I have been extremely unlucky with grains or somehow every one of my cultures has had the same exact contam. Including two different syringes from Myyco.
I vent my pressure cooker for 15 minutes and I PC at 15-17 psi for 90minutes with the millet and 2 hours with the oats. I've also tried 2.5 hours with oats to no avail.
Here are some photos from throughout my time growing:
 First Flush(cased the sub with extra coir when it was nearly fully colonized)
 Example of one of the first jars
 A plate from around the same time
 2nd or third tub
 2nd flush from this tub
 Jar from around this time. This one sat colonized for a few weeks before it looked this bad. When I spawned it looked better.
 Partially colonized jar. They don't start looking bad until after a shake or two and around 90% colonization.
 Next tubs start progressively getting worse

 The fruits on this tub were all stubby/stunted
 Last tub I spawned. Sub contam'd before it could pin. Lost hope for a bit. Note: Never noticed any other contam's in any jars. Throughout this time it's all been the same symptoms.
 Latest millet jar. inoculated with liquid culture from Myyco. Straight from the syringe. Same bacterial sour smell. Just spawned to bulk today anyway.
Edited by BExa (08/21/23 01:27 AM)
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Kmacmo
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: BExa]
#28441216 - 08/21/23 01:17 AM (1 year, 4 months ago) |
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The grain tek your using doesn't show you how dry the grains need to be and from your pics it does look like the grain has moisture on them. They should be completely dry on the outside. I don't use millet but Rye grains. Once I've boiled them I let them sit on kitchen paper spread out for a couple hours untill dry on the outside.
The moisture in the jar will settle at the bottom after a couple days and that's where the bacteria will thrive.
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10ftTall
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: Kmacmo]
#28441840 - 08/21/23 02:13 PM (1 year, 4 months ago) |
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What about trying BRF instead of grains?
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Crackatoa
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: 10ftTall] 2
#28441857 - 08/21/23 02:29 PM (1 year, 4 months ago) |
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We're going to get you through this brother. While that millet colonizes. Let's start back at step 1. When you pour your plates, do you let them sit for a week or so before using?
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mistermiyagi
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: BExa] 1
#28441882 - 08/21/23 02:48 PM (1 year, 4 months ago) |
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I believe those filter paper stickers are pretty suspect. I’ve been fighting my way to consistent axenic spawn and I’m pretty sure those filters were one of the contributing factors. For one, try breathing through them- almost impossible unlike polyfill or real sfd’s. That may or may not be related, but I often noticed what looked like signs of bacteria on grains towards the top of my jars.
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Crackatoa
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: mistermiyagi]
#28441884 - 08/21/23 02:53 PM (1 year, 4 months ago) |
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I use SFD stickers but cover them with 2 layers of MP tape.
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tryptkaloids
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: Crackatoa]
#28441889 - 08/21/23 03:09 PM (1 year, 4 months ago) |
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Are you venting your pc?
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Mycolorado
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: tryptkaloids]
#28441901 - 08/21/23 03:20 PM (1 year, 4 months ago) |
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Your first tubs sound pretty successful. Were you working in front of the merv13 like a hepa or just moving air around the room you were working in? Both would cause problems. I’d quit using the vendor LC and get back to using clean cultures produced by you and inoculating jars in the sab with no fans running and change your lid filters.
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phenyl
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: mistermiyagi]
#28441915 - 08/21/23 03:27 PM (1 year, 4 months ago) |
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Those jars all look crazy nasty for A2G unless something else is getting in them. You're not doing LC are you? Testing your SAB works properly with a plate exposure test and observation over a week or two is a good idea if you haven't already - they're not quite as independent from the environment as some people here assume. Mold is easy to settle but aerosols can stick around for a long time.
I'll tell this story a thousand times but after moving to a new apartment literally everything I made that was exposed to air became contaminated with bacteria. Nothing kept sealed developed the same issue. No change in protocols at all. Same equipment. FFU from Amazon completely fixed it. Sometimes it's not you but the shit in the air not settling even after an hour. In retrospect I suspect it was caused by a dryer with a condenser box blowing gross water into the air.
Quote:
mistermiyagi said: I believe those filter paper stickers are pretty suspect. I’ve been fighting my way to consistent axenic spawn and I’m pretty sure those filters were one of the contributing factors. For one, try breathing through them- almost impossible unlike polyfill or real sfd’s. That may or may not be related, but I often noticed what looked like signs of bacteria on grains towards the top of my jars.
The self-adhesive ones? Used several, all successfully. The smooth ones that come on blue plastic backing tend to need replacing more often than the nicely textured ones.
-------------------- The fool who persists in his folly will become wise.
Edited by phenyl (08/21/23 03:51 PM)
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mistermiyagi
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: phenyl]
#28441947 - 08/21/23 03:55 PM (1 year, 4 months ago) |
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Quote:
phenyl said: The self-adhesive ones? Used several, all successfully. The smooth ones that come on blue plastic backing tend to need replacing more often than the nicely textured ones.
Yeah the textured self-adhesive ones. When my jars got to almost full colonization it often looked like there was a patch of grains which refused to fully colonize under the filter. I can’t say definitively whether the filters were at fault though.
I think I’ve seen you recommend transferring to water agar as a regular part of the process, I’ve just done this for the first time and will continue to. I’m starting to think that contaminated vendor LC has also been a factor and it can be really hard for the untrained eye to spot.
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hazyhorse
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: BExa] 1
#28441956 - 08/21/23 04:07 PM (1 year, 4 months ago) |
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been having a mystery bacteria problem myself for awhile now, looks perfect up to 90% then goes to shit at full colonization. never get any other molds or anything on my plates or in my jars. shit is super frustrating to troubleshoot. i hope you get it figured out soon
one tip i’ve got in terms of spawning iffy grains is sticking to lower sub ratios (closer to 1:1 sub:spawn), prepping coir on the drier side of field cap, & giving plenty of FAE from the get go. might help ya get some better flushes out of your bacterial stuff while you’re figuring it out. those early tubs looked pretty banger all things considered, sucks to see your results get worse & worse like that
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phenyl
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: mistermiyagi]
#28441962 - 08/21/23 04:19 PM (1 year, 4 months ago) |
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Quote:
mistermiyagi said:
Quote:
phenyl said: The self-adhesive ones? Used several, all successfully. The smooth ones that come on blue plastic backing tend to need replacing more often than the nicely textured ones.
Yeah the textured self-adhesive ones. When my jars got to almost full colonization it often looked like there was a patch of grains which refused to fully colonize under the filter. I can’t say definitively whether the filters were at fault though.
I think I’ve seen you recommend transferring to water agar as a regular part of the process, I’ve just done this for the first time and will continue to. I’m starting to think that contaminated vendor LC has also been a factor and it can be really hard for the untrained eye to spot.
The top layer of grains drying out can happen. I shake and test recovery for a few days before I spawn so it doesn't bother me anyway, but I've spawned plenty of dried out millet on top of bags with no problem too. There's a thread wherein someone skimmed the dried out top grains off bags and spawned them solo - fruited fine. Not considered an issue as far as I'm aware. Whether that's what you're experiencing is another thing but I'd not expect it to be a huge problem unless something else was wrong.
Bacterial cultures can be hard to spot and so I always do a cleanup. No more "huh I guess this culture was contaminated" and needing to dump master jars or LCs. I'm too evolved to play hide and seek with prokaryotes. In this thread I'd expect at least something to come out reasonably clean from a bunch of different cultures, at least if they're doing A2G. If they're doing LC it's a bit easier to mess up considering the relative growth rate of anything in there resulting from removing water as a limiting factor.
-------------------- The fool who persists in his folly will become wise.
Edited by phenyl (08/21/23 04:28 PM)
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BExa
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: Crackatoa]
#28442290 - 08/21/23 11:33 PM (1 year, 4 months ago) |
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I have used plates that have sat around for a bit for sure. I don't believe dirty plates aren't the issue. I actually have a stack that's about 7 months old that are still clean. Not sure I'll use them though, they're looking a bit dried out.
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BExa
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: mistermiyagi]
#28442293 - 08/21/23 11:37 PM (1 year, 4 months ago) |
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I have suspected the filters as well. That has been the only constant throughout my time growing. I've changed the cultures, spawn, jars and PC time. Have you tried the filter stickers from Microppose? They look a bit different than the ones I got from Amazon. Perhaps they're more like synthetic filter disk material. I think I'm going to get both stickers and SFD's from them.
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BExa
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: Mycolorado]
#28442295 - 08/21/23 11:39 PM (1 year, 4 months ago) |
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I worked in front of the filters like a flow hood. I also tested them using a clean plate exposed to the air for 30 seconds. Came out clean. They worked surprisingly well. Went back to SAB this latest run though because I moved and got rid of my fans.
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JXAllen
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: BExa]
#28442311 - 08/21/23 11:59 PM (1 year, 4 months ago) |
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If you're talking about a box fan with a MERV13 filter, that's creating turbulent flow, not laminar. Laminar flow flows in one direction. This creates a zone of clean filtered air, all flowing in one direction. Turbulent flow will blow air in unpredictable patterns. This can draw in contaminants from the surrounding, unfiltered air, off of your work surface and off of you. If that's what you're using, you're far better off sticking with your SAB. Working in front of a filtered box fan is less reliable than working in still, open air.
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Edited by JXAllen (08/22/23 12:01 AM)
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mistermiyagi
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Re: Two years now, unable to get clean spawn. 100+ jars bacterial. [Re: phenyl] 1
#28675884 - 02/25/24 06:16 PM (10 months, 16 days ago) |
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Quote:
phenyl said:
Quote:
mistermiyagi said:
Quote:
phenyl said: The self-adhesive ones? Used several, all successfully. The smooth ones that come on blue plastic backing tend to need replacing more often than the nicely textured ones.
Yeah the textured self-adhesive ones. When my jars got to almost full colonization it often looked like there was a patch of grains which refused to fully colonize under the filter. I can’t say definitively whether the filters were at fault though.
I think I’ve seen you recommend transferring to water agar as a regular part of the process, I’ve just done this for the first time and will continue to. I’m starting to think that contaminated vendor LC has also been a factor and it can be really hard for the untrained eye to spot.
The top layer of grains drying out can happen. I shake and test recovery for a few days before I spawn so it doesn't bother me anyway, but I've spawned plenty of dried out millet on top of bags with no problem too. There's a thread wherein someone skimmed the dried out top grains off bags and spawned them solo - fruited fine. Not considered an issue as far as I'm aware. Whether that's what you're experiencing is another thing but I'd not expect it to be a huge problem unless something else was wrong.
Bacterial cultures can be hard to spot and so I always do a cleanup. No more "huh I guess this culture was contaminated" and needing to dump master jars or LCs. I'm too evolved to play hide and seek with prokaryotes. In this thread I'd expect at least something to come out reasonably clean from a bunch of different cultures, at least if they're doing A2G. If they're doing LC it's a bit easier to mess up considering the relative growth rate of anything in there resulting from removing water as a limiting factor.
So I don’t spread misinformation, the Amazon adhesive filters weren’t my source of contamination. Compared to jars with more expensive sfd’s they perform the same and have held up fine to several cycles in the dishwasher.
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