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Hiei
8======><=====8


Registered: 01/23/08
Posts: 2,648
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No colonization AT ALL in 8 jars at day 19
#26680343 - 05/19/20 05:13 AM (3 years, 8 months ago) |
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Hello everybody!
I PF tekked 8 jars like I usually have done in the past 4 or 5 times, always with decent results.
It was 2 verm: 1 BRF ratio and full of water (if I squeezed the verm in my hand some very few drops would come out)
then I covered the jars with two foil layers tight with a rubber band and sterilized them.
then I inoculated only in the center and left the hole open to let gas exchange.
there was a dry verm layer on top of every substrate.
Now it's been 19 days at right temp (25-27 degrees)
and NONE of them has any trace of colonization yet.
the syringe was kept loaded in the fridge for a while.
did this ever happen to you? what can be the cause?
Thankssss
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Edited by Hiei (05/19/20 05:14 AM)
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Hiei]
#26680351 - 05/19/20 05:16 AM (3 years, 8 months ago) |
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It is a too long time to have something different from now on. No hope I believe. The problem is with the syringe probably, or you inoculate when it is too hot.
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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Sockadin



Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680357 - 05/19/20 05:19 AM (3 years, 8 months ago) |
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Yeah seems like a long time, but if you inoculated in the middle of the cakes you may still have a week before you see any results.
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680358 - 05/19/20 05:20 AM (3 years, 8 months ago) |
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If you freeze the syringe it dies generally, under 0 Celsius is not okay. But fridge is fine. Even 1 year is also not a long time, but I do not know how long you mean you keep them there.
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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Hiei
8======><=====8


Registered: 01/23/08
Posts: 2,648
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Hiei]
#26680362 - 05/19/20 05:22 AM (3 years, 8 months ago) |
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meh, I could see the black spores inside and shook it before inoculating, also I waited some time after warming the needle... 
it stayed for some months in the fridge loaded...
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Edited by Hiei (05/19/20 05:23 AM)
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Sockadin]
#26680364 - 05/19/20 05:23 AM (3 years, 8 months ago) |
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Quote:
Sockadin said: Yeah seems like a long time, but if you inoculated in the middle of the cakes you may still have a week before you see any results.
Yeah but even if the jars are too large, 5-6 days at most for spores to germinate, and after 13 days they have still not reached to the edges? I hope you are correct but I do not believe so.
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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Sockadin



Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680366 - 05/19/20 05:24 AM (3 years, 8 months ago) |
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I do. Spores can take weeks to germinate and when they do you might not even tell because they are microscopic.
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Hiei]
#26680368 - 05/19/20 05:26 AM (3 years, 8 months ago) |
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Quote:
Hiei said: meh, I could see the black spores inside and shook it before inoculating, also I waited some time after warming the needle... 
it stayed for some months in the fridge loaded...
You can see spores does not mean anything I believe. You cannot say either they are alive by just looking. Maybe as I stated, you inoculate while jars are still hot? no?
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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Apples in Mono
Not a puppet


Registered: 09/21/17
Posts: 3,240
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680369 - 05/19/20 05:26 AM (3 years, 8 months ago) |
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Quote:
WhiteWalker said:
Quote:
Sockadin said: Yeah seems like a long time, but if you inoculated in the middle of the cakes you may still have a week before you see any results.
Yeah but even if the jars are too large, 5-6 days at most for spores to germinate, and after 13 days they have still not reached to the edges? I hope you are correct but I do not believe so.
It isn't unprecedented for germination to take a month
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Sockadin]
#26680378 - 05/19/20 05:30 AM (3 years, 8 months ago) |
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Quote:
Sockadin said: I do. Spores can take weeks to germinate and when they do you might not even tell because they are microscopic.
I see your point, and as stated before, I hope you are right, so that our buddy can have some outcome from his/her effort.
My 1 year spores still took 5-6 days to germinate. The duration you mentioned are correct for the dry spores, but for spores in liquid I do not remember such a long time. For mushrooms, everything is possible so maybe, maybe.
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Apples in Mono]
#26680385 - 05/19/20 05:33 AM (3 years, 8 months ago) |
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Quote:
Apples in Mono said:
It isn't unprecedented for germination to take a month
So you believe, it is the case also for syringes? or just for dry spores?
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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Hiei
8======><=====8


Registered: 01/23/08
Posts: 2,648
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Hiei]
#26680387 - 05/19/20 05:35 AM (3 years, 8 months ago) |
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Ok then: I'm waiting for a week more, then if I still see nothing I will try to inoculate them again, as I think a possibility may be that the jars were still too hot, even if I dont think so. I let them cool down overnight, but in the morning I might have still given a go to the cookers to be sure. Then I waited some time and also while inoculating the last jars would have cooled down more... but I'm thinking maybe the water inside was still quite hot and I didnt notice by touching them on the outside?
we'll se.... thank you guys
PS: I also just tried to shake them and put one more foil over it without rubber band.
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Edited by Hiei (05/19/20 05:37 AM)
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Sockadin



Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680390 - 05/19/20 05:36 AM (3 years, 8 months ago) |
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Yeah. Especially if they were kept in the fridge. There are so many variables that we do not have control over. I would advise either start over or wait longer. Either is up to the OP.
The only real way to test the spore syringe is on agar.
Edit: I would not inoculate them again. You can rePc or make new jars and allow them to cool for 12 Hrs. Also keep your syringe out of the fridge for a good 12 hrs to come up to room temp.
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Hiei
8======><=====8


Registered: 01/23/08
Posts: 2,648
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Sockadin]
#26680396 - 05/19/20 05:38 AM (3 years, 8 months ago) |
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Yeah the spores were fridge temperature when I inoculated, as the syringe was taken out just before inoculating... may that be a factor?
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Sockadin



Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Hiei]
#26680400 - 05/19/20 05:40 AM (3 years, 8 months ago) |
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Could be.
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: Hiei]
#26680406 - 05/19/20 05:44 AM (3 years, 8 months ago) |
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Quote:
Hiei said: Yeah the spores were fridge temperature when I inoculated, as the syringe was taken out just before inoculating... may that be a factor?
You mean a heat shock? No, I don't think it is a problem.
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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WhiteWalker
Dr.



Registered: 04/28/19
Posts: 67
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680407 - 05/19/20 05:45 AM (3 years, 8 months ago) |
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I never wait for syringe to come to room temp.
-------------------- "A diamond is a chunk of coal that did well under pressure." -Henry Kissinger
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Apples in Mono
Not a puppet


Registered: 09/21/17
Posts: 3,240
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Re: No colonization AT ALL in 8 jars at day 19 [Re: WhiteWalker]
#26680417 - 05/19/20 05:53 AM (3 years, 8 months ago) |
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Quote:
WhiteWalker said:
Quote:
Apples in Mono said:
It isn't unprecedented for germination to take a month
So you believe, it is the case also for syringes? or just for dry spores?
Either way. I'd go ahead and start something else in the meantime anyway, but give those guys another week or so
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