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Offlineagricola69
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Registered: 05/09/16
Posts: 4
Last seen: 6 years, 8 months
Contamination time after time
    #23922182 - 12/13/16 10:31 AM (7 years, 1 month ago)

Hello guys,
I've got a problem with my jars.
I've tried many times to grow my mycelium but every time I have this contamination.
It looks kind of similar to wet-spot and it also smells bad.

I followed step by step with PF-tek tutorials.
Jars were covered with 2x foil, holes in the lid were taped.
2 hours in PC.
injection in glovebox, everything was sterile.

Question 1.
From my observation it might be have something with spore prints, because I left 2 jars after sterilization for 2 weeks and nothing happened inside, but after I inject spores (from syringe) contamination came again.
I have spore print from FSRE, is it bad?

Question 2.
If it is wet-spot, how it is possible that bacteria (Bacillus sp.) is inside syringe (considering that nothing happened to 2 jars) as normally they appears on grain/rice?

Question 3.
What kind of procedure I should use to get rid off this bacteria, throw out jars, sterilization in PC then throw out "cakes", something else?

Photos of my jars:







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OfflineHuskies
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Registered: 03/22/16
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Re: Contamination time after time [Re: agricola69]
    #23922196 - 12/13/16 10:36 AM (7 years, 1 month ago)

Spore prints have a tendency to not be perfectly clean, wet spots and bad smell would indicate bacteria, rotten fruit right? If so the culprit is the spores themselves, which have bacteria hidden in them.

Search for pastyplates on here, follow those instructions, use a tiny drop of the spores that you have and grown on the agar, transferring out good growth away from any bacteria pops up.

When you have a second or third transfer plate, let it grow out, then pressure cook a jar of water, then put the clean agar in there. SHake it a bunch, and you will have Liquid Inoculant, which has live mycelium in it, and hopefully free of contaminants. Use that for your jars.


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Offlineagricola69
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Re: Contamination time after time [Re: Huskies]
    #23922225 - 12/13/16 10:44 AM (7 years, 1 month ago)

Thank you for your replay. I'll do that! :smile:
Could you also answer to my third question? What should I do with jars?


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OfflineHuskies
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Re: Contamination time after time [Re: agricola69]
    #23922253 - 12/13/16 10:50 AM (7 years, 1 month ago)

It's been a while since I have done cakes, there is growth going on them, you are sure they are contaminated? The wet rice stuff doesn't look good, but was it like that before?

Are you following a good tek? Are you using enough vermiculite?

If you are sure they are bad, then toss them, although bacteria is somewhat salvageable compared to green mold. Empty them outside of the house, clean the jars well with a sponge and dishsoap and you should be good.


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I call them Huskies cause you tell them to go "Mush! Mush""


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InvisiblePirateSwazey
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Re: Contamination time after time [Re: agricola69]
    #23922258 - 12/13/16 10:51 AM (7 years, 1 month ago)

You can just empty them in the trash or outside. I run a compost pile for spent substrates and contaminated grains the yard that I give to the neighbors for their garden once a year. You can sterilize them before hand if you want... Although I wouldn't worry about it with bacteria, just pink or black molds.

Wash the jars with hot water and dish soap.


If you want to see less contamination in the future you have to start working with agar! I would advise you to be moving to grains and coir substrates, starting everything you grow on petri dishes.


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Offlineagricola69
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Re: Contamination time after time [Re: PirateSwazey]
    #23922318 - 12/13/16 11:13 AM (7 years, 1 month ago)

Thank you guys for answers,

@Huskies
Well, As you said it smells like rotten fruit and with everyday the growth of mycelium is harder to notice that's is why I guess it is a contamination.


@Huskies, PirateSwazey

I am going to start with agar, but to make sure.
Are you talking about this (with attachments in article)? https://www.shroomery.org/forums/showflat.php/Number/19208976


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OfflineHuskies
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Re: Contamination time after time [Re: agricola69]
    #23922339 - 12/13/16 11:19 AM (7 years, 1 month ago)

Yup, that's the one!

follow it to a T and you will be golden.

And yeah, in that case toss them. I was under the impression you thought the white mycelium was something else because it wasn't heavy in the rhizo area, but if it smells bad start over. The agar will put you back a few weeks but it will be worth it.

Maybe even pressure cook a few q-tips, wrapped individually in tinfoil those wrapped in a larger piece. Instead of using spores directly on the plate, inject some of the spores on the sterile q-tip, and swab that on the agar plates.

final edit: some people have problems with agar in the beginning, i haven't and that was because I went for agar powder instead of the bars. I imagine it's easier getting the measurements right with it.

This is the stuff I use, I get it at asian markets

https://images-na.ssl-images-amazon.com/images/I/41fwTH8avRL.jpg


Edited by Huskies (12/13/16 11:27 AM)


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Offlineagricola69
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Registered: 05/09/16
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Re: Contamination time after time [Re: Huskies]
    #23922390 - 12/13/16 11:41 AM (7 years, 1 month ago)

I've been in the shop and bought powder agar :smile:
Hope that everything will be ok and in the next few weeks I will grow health mycelium.
Thanks for your help! :smile:


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OfflineHuskies
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Re: Contamination time after time [Re: agricola69]
    #23922411 - 12/13/16 11:46 AM (7 years, 1 month ago)

Godspeed!


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I call them Huskies cause you tell them to go "Mush! Mush""


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