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OfflineTimeToHarvest
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Registered: 09/26/23
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Agar transfer -> Inert Jars
    #28484061 - 09/26/23 06:33 PM (3 months, 30 days ago)

Disheartened and confused by the total lack of growth in my grain jars.

For my second attempt at growing Yellow Oysters & Lion's Mane, I did the following:
1. Built a simple still air box: plastic box with arm holes, wiped with alcohol, bottom lined with a towel soaked in dilute bleach, and the inside sprayed with Ozium.
2. Inoculated agar trays with spores; trays showed strong growth and no contamination.
3. Sterilized 32oz jars of millet using a pressure cooker - 15psi for 90 minutes.
4. With a sterile scalpel, cut generous slices of colonized agar and added them to the jars.

Now, 10 days later, there is NO growth in the jars of any kind. I opened one of the jars and found only millet stuck to a dead agar cutlet.  Can anyone suggest where I went wrong?

Thanks for the help


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Offlinedeadmandave
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Re: Agar transfer -> Inert Jars [Re: TimeToHarvest]
    #28484195 - 09/26/23 08:21 PM (3 months, 30 days ago)

What is ozium? Why not use alcohol?

Lm mycelium is very fine, perhaps your eye missed it.

Did your jars have gas exchange?

If all else fails, try again.


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OfflineTimeToHarvest
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Re: Agar transfer -> Inert Jars [Re: deadmandave]
    #28484229 - 09/26/23 08:42 PM (3 months, 30 days ago)

Ozium is a brand of air sanitizer; I sprayed it through the arm holes after wiping the sides down with alcohol.

I'm certain I didn't miss mycelium - the jars contained loose grain only.

Each of the jars had two holes in the lid, each covered with a micropore filter sticker. The lids were covered with tin foil.

I'm not giving up, but I need to identify what I've done that's killed 20 out of 20 jars across 2 fast-growing species.


Edited by TimeToHarvest (09/26/23 08:44 PM)


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OnlineRockinRobot
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Re: Agar transfer -> Inert Jars [Re: TimeToHarvest]
    #28484383 - 09/27/23 04:37 AM (3 months, 30 days ago)

Ozium is NOT an air sanitizer. It is simply an air freshener. Save it for covering up the smell of your weed. SMH


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OfflineOm Namo Shivaya


Registered: 05/03/15
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Re: Agar transfer -> Inert Jars [Re: RockinRobot]
    #28484403 - 09/27/23 05:29 AM (3 months, 30 days ago)

Whats the temperature in the room? Have any pictures? Pictures can always be helpful on the shroomery, if you have any pics if the agar plates and the jars.

Did you let the jars cool to room temp before innoculating? If the grains are too hot that can kill the mycelium.


Edited by Om Namo Shivaya (09/27/23 05:30 AM)


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OfflineTimeToHarvest
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Re: Agar transfer -> Inert Jars [Re: RockinRobot]
    #28484438 - 09/27/23 07:01 AM (3 months, 30 days ago)

Thanks for the replies.

Perhaps there are different varieties of Ozium, but the can I used has "Air Sanitizer" in big red letters and "Reduces Airborne Bacteria" in big white ones.

The jars had all cooled to room temperature; I'd sterilized them over the previous 5 days. The jars were stored in a box like the one pictured below with temperature controlled by an aquarium heater, set to 76f. They were in an un-climate controlled part of the house when exterior temp dropped to ~50f (likely 10-15 degrees warmer where they were).

Here is the inoculation set-up.  With supervised help, jars were un-foiled, wiped with alcohol wipes, and passed through the arm holes. Using the sterile scalpel, I put slices of cultivated agar into each jar, then resealed and passed them back out to be covered and labeled.


Edited by TimeToHarvest (09/27/23 07:16 AM)


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OfflineOm Namo Shivaya


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Re: Agar transfer -> Inert Jars [Re: TimeToHarvest]
    #28484570 - 09/27/23 09:47 AM (3 months, 30 days ago)

When you say it's dropped to 50F, do you mean just at night? or has it been 50 the whole time? If your chamber is only 10 degrees warmer, at 60F, that could explain no growth. Any pictures of the jars ? preferably a picture showing the piece of agar in the jar.


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OfflineOm Namo Shivaya


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Re: Agar transfer -> Inert Jars [Re: Om Namo Shivaya]
    #28484602 - 09/27/23 10:38 AM (3 months, 30 days ago)

I've had this happen before with oats. I use to have no problem with organic oats, but starting a couple years ago I started having issues where nothing would grow on them.

I side-by-side have the same culture on oats and wheat and the oats do nothing and the wheat colonize fine. The only guess I have is that the company started using a fungicide or something on their crops and it's persisting and killing the fungus, not sure what else it could be.


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Offlinedeadmandave
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Re: Agar transfer -> Inert Jars [Re: Om Namo Shivaya]
    #28484709 - 09/27/23 12:27 PM (3 months, 30 days ago)

Could be fungicide on the grains. Could be lack of ge from the micropore tape, perhaps the tape doesn't breathe as advertised.

No need for foil on lids during colonization, only exacerbates problems.

Try different grains, try different filter like polyfil, try not using an incubator. They will grow at 50f, just a touch slower.


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OfflineOm Namo Shivaya


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Re: Agar transfer -> Inert Jars [Re: deadmandave]
    #28484728 - 09/27/23 12:56 PM (3 months, 30 days ago)

Ah, yes iforget the species were talking about. When I mentioned no growth at 50-60 I meant for warm weather species


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OnlineRockinRobot
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Re: Agar transfer -> Inert Jars [Re: TimeToHarvest]
    #28484766 - 09/27/23 01:38 PM (3 months, 30 days ago)

Quote:

TimeToHarvest said:
Thanks for the replies.

Perhaps there are different varieties of Ozium, but the can I used has "Air Sanitizer" in big red letters and "Reduces Airborne Bacteria" in big white ones.

The jars had all cooled to room temperature; I'd sterilized them over the previous 5 days. The jars were stored in a box like the one pictured below with temperature controlled by an aquarium heater, set to 76f. They were in an un-climate controlled part of the house when exterior temp dropped to ~50f (likely 10-15 degrees warmer where they were).

Here is the inoculation set-up.  With supervised help, jars were un-foiled, wiped with alcohol wipes, and passed through the arm holes. Using the sterile scalpel, I put slices of cultivated agar into each jar, then resealed and passed them back out to be covered and labeled.





Why are you only wearing a glove on 1 hand?


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Offlinedeadmandave
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Re: Agar transfer -> Inert Jars [Re: RockinRobot]
    #28484819 - 09/27/23 02:26 PM (3 months, 30 days ago)

doesnt look like any gloves in the pic. gloves aren't necessary if you have clean hands and trim finger nails.


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OfflineTimeToHarvest
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Re: Agar transfer -> Inert Jars [Re: deadmandave]
    #28485059 - 09/27/23 06:04 PM (3 months, 29 days ago)

Thanks, all - really appreciate the ideas.

Temp-wise, the jars were in a box with an aquarium heater set to 76f; the box was in a room that got to 60f overnight.

I'll try again with different grain and without foil; given the oysters' total failure to jump from agar to millet, fungicide seems plausible. I have enough jars to try some with different filters and some outside the incubator.

This time, too, I'll remember to take pictures of the agar before inoculation.


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