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Offlinemanlord
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Massive Mold problems...Any1 experienced w/ Ozone?
    #27748347 - 04/23/22 04:23 PM (2 years, 8 months ago)

Hey guys, I've been having problems with my agar work for months now. EVERY strain I work on ends up with the exact same intertwined white mold, I suspect it is cobweb and/or verticillium (fungicola whatever).

Last night I checked in on my still air box, and noticed I'd left my Erlenmeyer flask sitting in it for a couple of days, open. All I saw was the white mold, and a couple of brown spots. The contamination (and the ratios of white mold vs brown), was exactly dead-on when compared to what I've been experiencing with my plates. They ALL end up getting the white mold, and some get brown, but way more white.

So I'm 100% convinced that by running a few rounds of tubs that had white mold from the get-go, I pretty much infested my house with mold. To the point that every plate I work on ends up with it. Whether I use a flowhood, an SAB, it doesn't matter. The problem actually gets WORSE with the transfers, even when I get a decent looking piece of growth on a germ plate, it ends up looking less rhizo and more moldy as I work on the culture.

The mold problem seems to be so bad that it gets into every plate I work on. No-pours work better, but still contam when I try to xfer from germ plates.

SO, I've purchased an Ozone generator. It's a cheap on off Amazon, about $100 and produces 20,000mg/hr. I ran it a couple weeks ago in my sterile room that I do culture work in. Turned it on for about 4-5hours, then aired the room out. That did not work. Probably because the rest of the house is still heavily coated in mold spores.

I've decided that I'm going to rent an airbnb or a hotel room for 2-3 nights, and run the ozone generator to kill all mold spores in the house. It's a 3bedroom house with a living and dining room and kitchen. All 1 floor though.

^^^^^Why I'm using Ozone ^^^^^


TLDR;

Anyone have experience using Ozone to rid a house of mold? What do you suggest, run it for 3-4 hours per room (That would amount to like 24 hours total)? Then once I'm done turn on fans and open windows for 6 hours or so? Should I have the central air on? And if so, should I put the generator near the air intake for a while?

I've seen conflicting info on whether to have AC on or not, how long is necessary (per room, and overall) to kill ALL mold spores, etc. Has anyone ever done this successfully, after having been unable to escape mold problems on agar?

Also I move out in a few weeks, I don't want to have that super heavy "clean"/bleachy smell to be too strong in the house. Is running it for 24 hours (3-4hours per room) going to be too much? I want to kill ALL mold, but I don't want the house to be smelling crazy and lose my security deposit etc.

Edited by manlord (04/23/22 04:24 PM)

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OfflineWeavieWonder
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: manlord] * 2
    #27748373 - 04/23/22 04:47 PM (2 years, 8 months ago)

Quote:

Man lord said:
SO, I've purchased an Ozone generator. It's a cheap on off Amazon, about $100 and produces 20,000mg/hr. I ran it a couple weeks ago in my sterile room that I do culture work in. Turned it on for about 4-5hours, then aired the room out. That did not work. Probably because the rest of the house is still heavily coated in mold spores.




Yeah that’s not gonna work. The world is covered in mold and bacteria spores. You come in the house everyday covered in them. General upkeep and cleaning of your home is more than sufficient. It’s possible you’ve got some serious deficiencies in your sterile technique. Even in a dingy dungeon, the proper use of a flow hood or SAB will give success.

Questions

1. Is the flowhood homemade? Pics?
2. When’s the last time you changed the filters?
3. Describe your technique when working with agar and grains.

Quote:

man lord said:
So I'm 100% convinced that by running a few rounds of tubs that had white mold from the get-go, I pretty much infested my house with mold. To the point that every plate I work on ends up with it. Whether I use a flowhood, an SAB, it doesn't matter. The problem actually gets WORSE with the transfers, even when I get a decent looking piece of growth on a germ plate, it ends up looking less rhizo and more moldy as I work on the culture.




Mold can be very difficult if not impossible to clean up on agar if you don’t catch it early. Sometimes prints and syringes are dirty af and are unworkable. Try getting some new prints to work with. Buy a few at a time in hopes that at least one is clean enough to work with.

If I’ve got mold on plates and can’t shake it after a couple of transfers, then I throw them away and start over. It sucks, but it’s better to do that then go through all the motions with dirty inoculant and have your projects shit the bed.

Edited by WeavieWonder (04/23/22 04:55 PM)

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OfflinePBJ710
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: WeavieWonder] * 1
    #27748391 - 04/23/22 05:06 PM (2 years, 8 months ago)

:whathesaid:

Ozone won't help you.  It's likely something in your sterile technique causing the root issues.  How about some pictures of your agar plates/jars/tubs?

Have you ever tested your FH with a lighter test to make sure they are functioning correctly?  Do control plates contam after being open if you don't make transfers to them?

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Offlinehazyhorse
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: WeavieWonder]
    #27748398 - 04/23/22 05:10 PM (2 years, 8 months ago)

are you using your SAB in a room with still air? an SAB won’t do shit for you if there are air currents in the room. like weavie said, SABs work with 100% success if used properly. you need to make sure there aren’t drafts, open vents, fans, AC etc. running. i promise you you don’t have some crazy spore load that no one has ever encountered before. there are trillions of spores in the air at all times. running ozone or whatever & then opening the windows isn’t gonna do anything to change that. if the air is truly still, spores don’t get blown around. either there’s some sort of draft running when you use your SAB, or your technique is off. you can’t really just leave your SAB up in the living room & expect it to work; you need to guarantee the air is still. more details of your actual process & pictures of your set up & plates would probably help


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Offlinedyel
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: PBJ710]
    #27748400 - 04/23/22 05:12 PM (2 years, 8 months ago)

post pics of seid mold good sir

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Invisibleredcrow

Registered: 03/25/22
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: manlord]
    #27748401 - 04/23/22 05:13 PM (2 years, 8 months ago)

Forget the ozone. Way back in '68 when I was just staring out as an RN, ozone was commonly used in hospitals to "sterilize" rooms that had been used by TB patients. Everyone and their uncle swore by it.

Then in '73, a study came out of NIH tracking hospital acquires cases of TB. The conclusion was that the ozonizers in use at the time (which put out far  more O3 than the paltry 20K mg/hr your unit does) didn't even kill vegetative staph aureus, let alone anything capable of producing spores.

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Offlinemanlord
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: manlord]
    #27748932 - 04/23/22 11:13 PM (2 years, 8 months ago)

I turn the central air off and have the vent to that room blocked. Some air might still leak under the door, which I have towels blocking, but other than that the air is still.

Test jars and plates left out in front of the flowhood didn't contam. But I may have been dealing with a mycoparasite at that point, that didn't germinate by itself on my agar.

But I ran like 20 tubs in a room right down the hall that all had mold. And that room had a fan going in it the entire time. I did 2 cycles like this. And since then I've been getting this mold in all my plates once I work on them.

My no-pour germ plates often show some decent growth, but as I transfer, and expose the plates to the environment, the rhizo growth goes away and gets overtaken by mold. I spray everything so heavy with iso, I never move above a plate I work right in front of the flowhood, etc etc. I shower and brush teeth, mouthwash, new clothes, all that.

Flowhood and SAB both have the issue. And Ive done hundreds of plates. Even if my sterile tek was off, I still don't think EVERY single plate would contam with the exact same mold, which happens to be growing just down the hall being sprayed all over the house with a fan. 

Here's pics of a bunch of plates. Some recent and some older. I've had some better looking plates at some points (not shown here), but they all end up moldy. Ever since I ran a big grow with mold and let it go to fruiting multiple flushes, I've had these issues.



And my flowhood is one that I purchased. I haven't checked the filter, but once I considered how moldy my house probably was, I switched to an SAB to just try to keep things still and not pull in dirty air from the hallway into the room I'm working in.

I understand that mold and bacteria are everywhere, but I'm not in a dirty dungeon, I'm literally within 10 steps of a room where there have been like 60 monotubs over a 3-4month period running with 100% mold intertwined with the myc. And a fan running in that room. I mean I'm not saying outside isn't dirty, houses arent dirty, etc. But I've been blasting my whole house with vert/cobweb for months on end...and that's the contam I seem to always get on agar. I've already gone down the sterile tek, clean the house, no-pours, warm-pours, I've already tried all that. So I'm going to work on this theory because it seems to be connected. Same contam, showed up right when I started running all those mold tubs.

I just don't think my sterile tek is causing every single plate to contam with mold. Everything sprayed with iso, I have gloves and arm sleeves, torch for sterilization, alcohol pads, etc. Why would I just constantly have this 1 type of mold on my gloves/sleeves that were just iso'd, over months of agar work? Of course my work space is clean and wiped with iso, I don't think I'm missing 1 type of mold every single time.

Edited by manlord (04/23/22 11:24 PM)

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OfflinePBJ710
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: manlord]
    #27748950 - 04/23/22 11:32 PM (2 years, 8 months ago)

Sorry, but most of those cultures look pretty bad IMO.  Plate 3 is the only one I would take a transfer from out of the entire group and then only that little cluster of rhizo growth on the very bottom edge. 

Once organisms get intermingled, it becomes much more difficult to seperate them.  It can be done, but its often alot easier/quicker to just start over from spores and get a 100% axenic culture to start with.

What recipe are you using for your agar?

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Offlinemanlord
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: PBJ710]
    #27748968 - 04/23/22 11:50 PM (2 years, 8 months ago)

I've used all different recipes. BRF agar, Malt extract, Malt/yeast, PDA. I've done all different ratios, even water agar to see if I could outrun the mold.

Yeah I don't even have any of those plates rn they are all old pics. But that's what I'm dealing with, every time. Not trich, or bacteria, like specifically that cobweb or verticillium whichever it is. I really don't think it's my sterile tek because that seems way too specific to have the same thing in my tubs and my agar, every single time and never anything different. My environment has to have a heavy sporeload for this mold.

That's why I'm asking about ozone/ any other way to lessen the sporeload in a home.

Edited by manlord (04/23/22 11:50 PM)

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Offlinehazyhorse
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: manlord]
    #27748992 - 04/24/22 12:22 AM (2 years, 8 months ago)

damn man, that sucks. i hope you get an answer soon. i'd be frustrated with those plates too.


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

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OfflinePBJ710
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: hazyhorse]
    #27749203 - 04/24/22 07:03 AM (2 years, 8 months ago)

I feel your pain.  I lost a TWC culture to similar intermeshed shit like that once.  It might have been savable, but since the culture wasn't incredible I didn't even bother and just snagged a new one from spores.

Unfortunately Ozone is like chemo - it damages every living thing it comes in contact with, not just the undesirable contams.  It's already too late if you are relying on 'sterile' air to maintain the health of your cultures. 

Usually I've seen stuff like that when I insist on using a dirty plate to make a transfer from (ya know that logic - it's only a tiny mold colony on the side of the plate :wink:).  Sometimes those little colonies dump enough spores ON the mycelium in the plate that they start growing together.  I know I have black mold in the wall behind where I keep my tubs yet I don't have problems with it meshing with my cultures.  I do occasionally see a bag of grain fail at inoculation, but that's normally due to being a clutz when sealing bags and breaking my sterile technique.

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InvisibleLanguidNoodle
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Re: Massive Mold problems...Any1 experienced w/ Ozone? [Re: manlord]
    #27807472 - 06/05/22 08:00 PM (2 years, 7 months ago)

Dealing with the same thing. Will try to amend and keep posted

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