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OfflineFonzee
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Constant contaminations
    #26909090 - 08/31/20 05:09 AM (3 years, 5 months ago)

Hi shroomerites,

I'm having constant problems with contams. I work with a good flow hood, clean my cultures on agar etc. Even got an AC recently.
But still, I'm getting a lot of contams. Now its mainly coweb mold (I guess), and I see it starting at random grains after I put the agar \ G2G in the sterilized jars, rarely before. I would have guessed its my hood teks but the agar work is pretty good.

Any idea how to avoid these contams?



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OfflineA.k.aM
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Re: Constant contaminations [Re: Fonzee]
    #26909163 - 08/31/20 06:36 AM (3 years, 5 months ago)

If it’s showing up all over the jar at the same time it’s probably from whatever you use to inoculate. I’d guess something got in the original jar and then blew up when it got g2g.


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OfflineFonzee
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Re: Constant contaminations [Re: A.k.a]
    #26909169 - 08/31/20 06:45 AM (3 years, 5 months ago)

The jar in the picture was inoculated with an agar slice


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Offline3SIXTY5
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Re: Constant contaminations [Re: Fonzee]
    #26909173 - 08/31/20 06:46 AM (3 years, 5 months ago)

What's your process and prep of grain spawn consist of?

How did you go about getting your culture? MS --> Agar --> Agar --> Grain? How many transfers, etc?

What are the specs on your blower a d filter, can you post pics of your flow hood too?


Edited by 3SIXTY5 (08/31/20 06:47 AM)


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OfflineA.k.aM
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Re: Constant contaminations [Re: 3SIXTY5]
    #26909178 - 08/31/20 06:48 AM (3 years, 5 months ago)

So it’s on the agar or got in when you dropped the wedge.

Basically if it’s not on the very top of the grain then it’s usually been in there the whole time and didn’t just sneak in through a faulty filter or something like that.


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Offline3SIXTY5
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Re: Constant contaminations [Re: A.k.a]
    #26909184 - 08/31/20 06:51 AM (3 years, 5 months ago)

Quote:

A.k.a said:
So it’s on the agar or got in when you dropped the wedge.

Basically if it’s not on the very top of the grain then it’s usually been in there the whole time and didn’t just sneak in through a faulty filter or something like that.




This is most likely the case, let's focus on your flow hood first, specs of blower/filter, and a picture of your hood would be ideal.


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OfflineFonzee
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Re: Constant contaminations [Re: 3SIXTY5]
    #26909395 - 08/31/20 10:02 AM (3 years, 5 months ago)

Quote:

3SIXTY5 said:
What's your process and prep of grain spawn consist of?

How did you go about getting your culture? MS --> Agar --> Agar --> Grain? How many transfers, etc?

What are the specs on your blower a d filter, can you post pics of your flow hood too?



I use wheat grain, no soak, cook to a rolling boil and let it cook until the grain are kind of soft but not mushy.
I then steam dry them on a drying rack with a fan, load jar 1/2 to 3/4 way and PC for 90 minutes.

I'm working from spore \ live culture to agar, and usually go for a minimum of 5 transfers, but most cultures I have went through more transfers. Then I drop quarter to half a plate on the grain and shake.

I'm not really sure about the specs, I bought the whole thing as a complete unit from a reputable seller. I got a feeling it is too strong for laminar flow though.
I have the hepa specs, but not the fan specs.

Thanks for helping!

Quote:

A.k.a said:
So it’s on the agar or got in when you dropped the wedge.

Basically if it’s not on the very top of the grain then it’s usually been in there the whole time and didn’t just sneak in through a faulty filter or something like that.



Quote:

3SIXTY5 said:
That makes sense. Maybe I should clean the outer side of my jars with alcohol before working with them.
The contams could have got from the outer side of the jars and spread around when I shook them.

Thanks for helping!

Quote:

A.k.a said:
So it’s on the agar or got in when you dropped the wedge.

Basically if it’s not on the very top of the grain then it’s usually been in there the whole time and didn’t just sneak in through a faulty filter or something like that.




This is most likely the case, let's focus on your flow hood first, specs of blower/filter, and a picture of your hood would be ideal.



If needed I would gladly send a pic.


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Offline3SIXTY5
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Re: Constant contaminations [Re: Fonzee]
    #26911242 - 09/01/20 06:31 AM (3 years, 5 months ago)

Quote:

Fonzee said:
If needed I would gladly send a pic.




How close do you keep your dishes to the filter face while you're doing you culture work?

Might as well send throw a pic up if it's not too much of a hassle.

I'd be willing to bet your issue is coming from your dishes based on what you've told us so far, one thing you could do that would eliminate the flow hood as the issue would be to build an SAB and just do a few jars in the SAB, if they still contaminate the same way then you know it's your agar.

If you're using wheat kernels, you should do a soak, for wheat kernels you shouldn't do longer than a 14 hour soak, and make sure you soak in cold water, if you soak in warm or hot water they will start to ferment within like 8 hours. I had much higher contamination rates when I wasnt soaking, Soaking isn't just to help hydrate the grains, it's to give the endospore a chance to germinate before pressure cooking, if they haven't germinated before the PC there's a higher chance that they will survive the PC cycle.

A couple years ago, I was having constant issues with trich, but for me it wouldn't contaminate until my substrates were nearly done colonizing. I thought I wasn't pasteurizing properly, then I thought it was my grain prep, I thought my house just had a really high trich spore load, lol I was driving myself nuts, I made probably 30-40 monotubs before I realized what the issue was, and it was my culture getting too old so the mycelium was just too weak to fight off Contamination, not saying that's whats happening with you, but just something to think about.


Edited by 3SIXTY5 (09/01/20 06:33 AM)


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OfflineFonzee
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Re: Constant contaminations [Re: 3SIXTY5]
    #26913014 - 09/02/20 12:17 AM (3 years, 5 months ago)

Quote:

3SIXTY5 said:
How close do you keep your dishes to the filter face while you're doing you culture work?

Might as well send throw a pic up if it's not too much of a hassle.

I'd be willing to bet your issue is coming from your dishes based on what you've told us so far, one thing you could do that would eliminate the flow hood as the issue would be to build an SAB and just do a few jars in the SAB, if they still contaminate the same way then you know it's your agar.

If you're using wheat kernels, you should do a soak, for wheat kernels you shouldn't do longer than a 14 hour soak, and make sure you soak in cold water, if you soak in warm or hot water they will start to ferment within like 8 hours. I had much higher contamination rates when I wasnt soaking, Soaking isn't just to help hydrate the grains, it's to give the endospore a chance to germinate before pressure cooking, if they haven't germinated before the PC there's a higher chance that they will survive the PC cycle.

A couple years ago, I was having constant issues with trich, but for me it wouldn't contaminate until my substrates were nearly done colonizing. I thought I wasn't pasteurizing properly, then I thought it was my grain prep, I thought my house just had a really high trich spore load, lol I was driving myself nuts, I made probably 30-40 monotubs before I realized what the issue was, and it was my culture getting too old so the mycelium was just too weak to fight off Contamination, not saying that's whats happening with you, but just something to think about.



I keep my cultures as close as possible to the hood, without touching it, while working.
I'll get some dish pics soon, hope that would be helpful for the diagnosis.

I have an issue with soaking. I can't really set my times right and a lot of times I end up not cooking my soaked grains on time, like 2 days later, and I lost a LOT of grain to that. I used to soak for a while, but I didn't notice any real difference once I stopped to.
At our weather, 8-10 hours soak is plenty. We get around 32°C constantly. After 16-18 hours they already start to germinate.

I had a lot of faulty tubs too lately, but I thought it was the weather so I got an AC in the room. Again, I'll post some agar pictures. Might even start a thread for it as I have quite a few of them...

Thanks for helping!



(Sorry for the mass, I still need to inoculate the bags and jars so they are clogging up the space. Got another 7 bags still in the PCs...)


Edited by Fonzee (09/02/20 12:18 AM)


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