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InvisibleWay
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Registered: 01/14/23
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Re: Recognizing and dealing with contamination [Re: rumfor69]
    #28577508 - 12/10/23 05:56 PM (1 month, 17 days ago)

Quote:

Smellyhobbit said:
Quote:

Way said:
Your sterile work is usually spot on. You're not a noob and you know what you're doing. A2A and A2LC working without failure shows your transfers are  fine.

You are running multiple different cultures so it can't be a culture related issue.

That leaves grain prep and sterilization right?

Have you left a control jar running without inoculating to see if it succumbs as well? If it does, it is definitely the PC giving up the ghost or an issue with the grain prep. If it doesn't, you've started doing something different during A2G that is causing the issue I would assume.



I’ll prep some grain today and set aside a couple of control jars. Give them a vigorous shake every week.

No different procedures in A2G. If it’s grain prep, why is the LC working? And I had a round of great success first time I tried this new grain prep followed by total A2G failure in subsequent batches. I’d say 2 batches of grain performed amazingly then nothing after.

Do you think it could be a grain QUALITY issue? I mean 2 hours in the PC should be sufficient, and even then the LC shouldn’t work if it’s a quality or prep issue, right?

I’m about to burn the apartment down and rebuild it out of stainless steel and glass so it can be flame sterilized




Honestly I read wrong as well and didn't realize LC to grain was working. Okay, so we know your cultures are good and your grain is good then. That leaves some gap in your sterile practices causing it that you have somehow been getting away with for a year up until now? I'm stumped as well. I hope you get this resolved soon though because that's a major bummer.


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OfflineSmellyhobbit
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Re: Recognizing and dealing with contamination [Re: Way] * 1
    #28577524 - 12/10/23 06:02 PM (1 month, 17 days ago)

I usually use my mouth to suck some agar off the plate, chew it to form a sort of slurry, and then use a cough to distribute it evenly through the grain jar.

Anything in that process strike you as bad sterile technique I could improve on?


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InvisibleBigwormS
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Re: Recognizing and dealing with contamination [Re: Smellyhobbit]
    #28577552 - 12/10/23 06:18 PM (1 month, 17 days ago)

Are you scraping the agar off the scalpel on the rim of the jar, do you shake it off, or do you tap the scalpel on the rim to knock the agar in to the jar? Are you just taking the colonized portion or do you leave an uncolonized edge on your agar? 
I'm assuming the LC is colonizing faster then the a2g? Do you do all the different inoculation methods in the same sitting? Which one do you do first, if you do them at the same session.


Edited by Bigworm (12/10/23 06:21 PM)


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OnlineSupaThaRipper
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Re: Recognizing and dealing with contamination [Re: Smellyhobbit] * 2
    #28577554 - 12/10/23 06:20 PM (1 month, 17 days ago)

Quote:

Smellyhobbit said:
I usually use my mouth to suck some agar off the plate, chew it to form a sort of slurry, and then use a cough to distribute it evenly through the grain jar.

Anything in that process strike you as bad sterile technique I could improve on?




No that should all work. Unless you’ve been sick lately?


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Offlinehazyhorse
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Re: Recognizing and dealing with contamination [Re: Smellyhobbit] * 1
    #28577571 - 12/10/23 06:32 PM (1 month, 17 days ago)

Quote:

Smellyhobbit said:
I usually use my mouth to suck some agar off the plate, chew it to form a sort of slurry, and then use a cough to distribute it evenly through the grain jar.

Anything in that process strike you as bad sterile technique I could improve on?




spit is sterile, as we all know. but are you brushing your teeth/flossing before hand? you could be adding some residual unsterilized nutrition if you have anything gunked up in your molars

Quote:

rumfor69 said:
Ohhh yeah I see that now. Dunno where my mind was lol I'll just go look at stuff in the corner now




don’t worry bud we’ll get you a bottle of glue to sniff while you’re over there :inlove3:


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OfflineSmellyhobbit
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Re: Recognizing and dealing with contamination [Re: Bigworm]
    #28577574 - 12/10/23 06:36 PM (1 month, 17 days ago)

Quote:

Bigworm said:
Are you scraping the agar off the scalpel on the rim of the jar, do you shake it off, or do you tap the scalpel on the rim to knock the agar in to the jar? Are you just taking the colonized portion or do you leave an uncolonized edge on your agar? 
I'm assuming the LC is colonizing faster then the a2g? Do you do all the different inoculation methods in the same sitting? Which one do you do first, if you do them at the same session.




I usually have the agar wedge on the blade where tilting it will cause it to fall into the jar. If that doesn’t work I just do what I have to. Just give it a little momentum until it falls.

The LC is done in a little less than half the time of a normal A2G jar.

I don’t really discriminate on what I do inside the box. It’s a space issue. If there’s room to do like 2 LC inoculations and 2 A2G inoculations I will, otherwise I split it up between 2 sessions.

I have no set order for these activities. Whatever is closest to me at the moment. Probably LC first because I don’t have to get my blades ready right away.


--------------------
A Love Letter to New Growers
A Guide for New Growers
Growth 2023 - A Year In Review

Grow more shrooms. Eat more ass. :mushroom:



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OfflineSmellyhobbit
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Re: Recognizing and dealing with contamination [Re: SupaThaRipper] * 1
    #28577575 - 12/10/23 06:37 PM (1 month, 17 days ago)

Quote:

SupaThaRipper said:
Quote:

Smellyhobbit said:
I usually use my mouth to suck some agar off the plate, chew it to form a sort of slurry, and then use a cough to distribute it evenly through the grain jar.

Anything in that process strike you as bad sterile technique I could improve on?




No that should all work. Unless you’ve been sick lately?



I recently had a fungal infection in my mouth, but I’m having mold problems not fungus problems. I want fungus. I’m getting mold.


--------------------
A Love Letter to New Growers
A Guide for New Growers
Growth 2023 - A Year In Review

Grow more shrooms. Eat more ass. :mushroom:



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Onlinehuey.bluey
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Re: Recognizing and dealing with contamination [Re: Smellyhobbit] * 2
    #28577582 - 12/10/23 06:45 PM (1 month, 17 days ago)

My procedure is loading my scalpel into my Anus like a sharpie and launching it across my sab , into my open plate , meticulously separating a slither of hyphae as it's released on the receiving dish. I rarely have any problems with this technique.


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OfflineAspectOfTheCreator
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Re: Recognizing and dealing with contamination [Re: Bigworm] * 1
    #28577727 - 12/10/23 07:57 PM (1 month, 17 days ago)

Quote:

Bigworm said:
Are you scraping the agar off the scalpel on the rim of the jar, do you shake it off, or do you tap the scalpel on the rim to knock the agar in to the jar? Are you just taking the colonized portion or do you leave an uncolonized edge on your agar? 
I'm assuming the LC is colonizing faster then the a2g? Do you do all the different inoculation methods in the same sitting? Which one do you do first, if you do them at the same session.





Whats wrong with using the inner rim of the jar to aid in sliding the wedge off? The glass must be sterile. Grains make contact there when shaking the jar. What am I overlooking?


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Offlinerumfor69
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Re: Recognizing and dealing with contamination [Re: AspectOfTheCreator] * 2
    #28577826 - 12/10/23 08:43 PM (1 month, 17 days ago)

I used to use the slide off the blade with a little momentum technique but switched to inner jar glass swipe off cause it means less time getting the wedge on the blade right so the dish is open less longer.


Oooo...this glue is nice :celery:


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Offlinehazyhorse
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Re: Recognizing and dealing with contamination [Re: AspectOfTheCreator]
    #28577884 - 12/10/23 09:50 PM (1 month, 17 days ago)

unless there’s something i’m unaware of i don’t think there’s anything wrong with using the inner edge, like you said it’s sterile.


--------------------
you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================

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InvisibleBigwormS
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Re: Recognizing and dealing with contamination [Re: rumfor69] * 2
    #28577887 - 12/10/23 09:53 PM (1 month, 17 days ago)

I'm just looking for anything that might help Hobbit out. The glass on the inner rim should be fine, if there was no grain stuck in the threads to allow air to get sucked in while shaking it after PC cycles to loosen the grain before inoculation. The shaking agar off would cause air currents. The order would be maybe from moving so much in and out of a SAB might cause contamination to be sucked in to the SAB from air currents as well. This would mean that if the grain was always last it could be dirty air in the SAB. I was just trying to rule things out.

Now the LC is done in half the time which might be why it's not contaminating. If the agar takes double the time bacteria/mold might get a better foothold before the healthy myc does. The amount of agar that is uncolonized could be giving bacteria/mold, that's hiding in the grain, something really nutritional to grow on when the agar is introduced to the grain. Which might explain why the a2grain fails but LC is good. I'm no expert and I fail a lot too so take what I say with a grain of salt. It is just something to consider.
I had to sit and really think about my failures and looked at every possible contamination vector possible. These were just my thoughts when I did that. Figured it might help out a bit.


Edited by Bigworm (12/10/23 09:56 PM)


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Offlinerokafogtacsuka
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Re: Recognizing and dealing with contamination [Re: Bigworm]
    #28577982 - 12/11/23 01:28 AM (1 month, 17 days ago)

Hey there,

I have some greyish/blueish stuff in my brown rice bags. Is that mold? Thanks for checking!





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Offlinehazyhorse
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Re: Recognizing and dealing with contamination [Re: Bigworm]
    #28578002 - 12/11/23 02:23 AM (1 month, 17 days ago)

Quote:

rokafogtacsuka said:
Hey there,

I have some greyish/blueish stuff in my brown rice bags. Is that mold? Thanks for checking!








that bag is bad, toss it

Quote:

Bigworm said:
I'm just looking for anything that might help Hobbit out. The glass on the inner rim should be fine, if there was no grain stuck in the threads to allow air to get sucked in while shaking it after PC cycles to loosen the grain before inoculation. The shaking agar off would cause air currents. The order would be maybe from moving so much in and out of a SAB might cause contamination to be sucked in to the SAB from air currents as well. This would mean that if the grain was always last it could be dirty air in the SAB. I was just trying to rule things out.

Now the LC is done in half the time which might be why it's not contaminating. If the agar takes double the time bacteria/mold might get a better foothold before the healthy myc does. The amount of agar that is uncolonized could be giving bacteria/mold, that's hiding in the grain, something really nutritional to grow on when the agar is introduced to the grain. Which might explain why the a2grain fails but LC is good. I'm no expert and I fail a lot too so take what I say with a grain of salt. It is just something to consider.
I had to sit and really think about my failures and looked at every possible contamination vector possible. These were just my thoughts when I did that. Figured it might help out a bit.




for sure, it's good to know the intricate details of someone's process to help troubleshoot, & it's gonna take thinking from a lot of different angles.

that being said, his agar plates don't ever get this mold on them & you'd think mold would still grow in the LC or get in the jars during an LC inoculation. like even a little mold would still cause failure if spores germinated in the grains right? i doubt the mycelium would colonize fast enough to avoid the other mold germinating. i guess that is a good question- when does the mold pop up after inoculating with an agar wedge?


--------------------
you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================

🅃 🄴 🄰 🄼    🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿


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OfflinePineappleBrains
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Re: Recognizing and dealing with contamination [Re: hazyhorse]
    #28579431 - 12/11/23 11:59 PM (1 month, 16 days ago)

I've got some concerns about both this monotub and a smaller shoebox I have with some contamination and looking for some confirmation.

This one in the center has a bit of green goin on.

It didn't really show up on camera but the left hand side of this pic has a slight green hue to it.

Here is an overall shot of the tub.

This is from the smaller shoebox. I took the picture because of the bit in the center but also see it to the left as well.

Also this bit in the center.

Here is an overall shot of the shoebox.


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Offlinerumfor69
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Re: Recognizing and dealing with contamination [Re: PineappleBrains]
    #28579606 - 12/12/23 06:43 AM (1 month, 16 days ago)

Looks like blue bruising to me


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InvisibleBigwormS
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Re: Recognizing and dealing with contamination [Re: rumfor69]
    #28579638 - 12/12/23 07:07 AM (1 month, 16 days ago)

:whathesaid:


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Offlinerumfor69
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Re: Recognizing and dealing with contamination [Re: Bigworm]
    #28579667 - 12/12/23 07:45 AM (1 month, 16 days ago)

That little center slimey blob might be some bacteria but it's nothing to worry about now. You're fruiting, you won fam, good job
:highfive:


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OfflineAspectOfTheCreator
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Re: Recognizing and dealing with contamination [Re: PineappleBrains]
    #28579686 - 12/12/23 08:14 AM (1 month, 15 days ago)

It wouldn't be flushing so well if it was contaminated.


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Offlinekengken
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Re: Recognizing and dealing with contamination [Re: AspectOfTheCreator]
    #28580504 - 12/12/23 08:22 PM (1 month, 15 days ago)

I have a couple of monotubs (27qt) with Penis Envy growing. Well...I'm still figuring things out so I ended up letting the substrate get way too dry and also was leaving the grow lights on 24/7, which I'm sure contributed to the dryness.

Both of the bins stalled out after getting a few pins. Tub #1 had some very dense pin "mounds" in the corners where there was probably more moisture. Once I figured out my mistakes I did some bottom watering, misted a bit, and in just the past few days have been doing only 12 hours of light. One bin really took off and got to the point where I harvested about 2oz once dried.

Question is, did I end up with some contamination or is what I'm seeing in the below pictures explained by the poor growing conditions? If contam do I need to toss everything I've harvested, and is there any salvaging them?

I'm thinking bin #1 could just be showing some bruising on the myc...but that might just be wishful thinking. And the orange/yellow myc in the second bin could be metabolites.

Bin #1




Bin #2




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