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OfflineZsuzsi
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Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
14 days, no mycellium in all jars but one
    #25889912 - 03/22/19 10:22 AM (5 years, 9 months ago)

Hello,

after reading tons on this excellent forum, I decided to start my own cultivation. I went full on pf tek / BRF cakes (and plan to use a shotgun fruiting chamber), firstly because I don't have a pressure cooker and secondly because it seemed the most likely way to succeed for a first culture and to familiarize myself with mushroom growth.

I inoculated 8 230ml (1/2 pint) jars almost exactly 14 days ago; they were sterilized using one of those vegetable steamers (used to cook vegetables or meat using steam), for two hours. I inoculated about 0.25cc of the spore suspension from the spore syringe in each of the four holes of every jars (so 1cc per jar), and the holes were afterwards covered with micropore tape. Inoculation was performed in a DIY still air box. The spore syringe is B+, and it's fresh (it had just arrived) and from a reliable source.

The exception is for two of the jars; which received almost 3cc each: the first one because I didn't measure the sensitivity of the syringe properly at first, the second because it was the very last jar and I pushed all of what was left in the syringe. Incidentally, the jar where there is mycelium growth is one of those two.

It has been 14 days; I tried to disturb the jars as little as possible. Only one jar so far shows signs of mycelium growth; I'd say it's about 20% colonized. The first signs of mycelium appeared at about 7 days after inoculation. The mycelium looks healthy, and there is no obvious contamination visible.

The other seven jars don't show any sign of contam; but they don't show signs of mycelium growth either. I have a few questions:

  • On the jar where there is growth, I have observed some condensation on the inside of the jar, specifically above where the mycelium is against the glass, and only where there is mycelium. I guessed this may be due to the slight heat produced by the mycelium itself growing; would that be correct ? should I be worried about the condensation ?

  • Is there a chance for the other jars to show some growth by now ? I think the main factor that could impact growth here is temperature; they're just sitting in a closet and I'd guess the average temperature is maybe around 2 or 3C less than ideal. Would lower temperature prevent growth at all ? delay it ? if so, by how much ? could attempting to move them to a slightly hotter place help, or hinder things at this point ?

  • Am I correct in assuming that the fact that there is no contam in any jar means I followed a proper procedure in terms of sterility ? I'd like to know because I have two syringes left, and I may plan on trying a grain/monotub based approach next, with a pressure cooker for sterilization.

  • How should I store the currently-unused spore syringes for long term viability ? there's sitting in the fridge inside a small carton box, is that OK ?


Thanks in advance !

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OfflineFun-G
Dr gonzo
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Registered: 03/05/19
Posts: 195
Last seen: 3 years, 11 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25889960 - 03/22/19 10:51 AM (5 years, 9 months ago)

What is your temperature at where you are incubating the jars?

Older spores can take up to 21 days to mate. The condensation is normal. Small evaporation can be expected.

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Offlineteladi
FUNKSOULBROTHER

Registered: 06/27/17
Posts: 1,189
Loc: South Africa Flag
Last seen: 1 year, 5 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25889974 - 03/22/19 10:56 AM (5 years, 9 months ago)

Quote:

Zsuzsi said:

  • On the jar where there is growth, I have observed some condensation on the inside of the jar, specifically above where the mycelium is against the glass, and only where there is mycelium. I guessed this may be due to the slight heat produced by the mycelium itself growing; would that be correct ? should I be worried about the condensation ?






A bit of condensation isn't much worry. You're correct in the reason why.

Quote:

Zsuzsi said:

  • Is there a chance for the other jars to show some growth by now ? I think the main factor that could impact growth here is temperature; they're just sitting in a closet and I'd guess the average temperature is maybe around 2 or 3C less than ideal. Would lower temperature prevent growth at all ? delay it ? if so, by how much ? could attempting to move them to a slightly hotter place help, or hinder things at this point ?






Either they haven't germinated yet, or there's nothing to germinate. The latter happens from time-to-time. You could re-inoculate now, or wait. Don't stress about the temperature too much.

Quote:

Zsuzsi said:

  • Am I correct in assuming that the fact that there is no contam in any jar means I followed a proper procedure in terms of sterility ? I'd like to know because I have two syringes left, and I may plan on trying a grain/monotub based approach next, with a pressure cooker for sterilization.






I wouldn't say you have completely sterilized those jars. The process you took was good enough for PF Tek, as it knocks contamination down enough so that mycelium can colonize and beat out any bad stuff. I'd suggest starting to learn how to do agar. Once you have some culture cleaned up on agar, you can keep that going for a very long time, without having to go back to spores.

Quote:

Zsuzsi said:

  • How should I store the currently-unused spore syringes for long term viability ? there's sitting in the fridge inside a small carton box, is that OK ?







Somewhere cool and dark. In the fridge is fine.

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OfflineZsuzsi
Stranger
Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
Re: 14 days, no mycellium in all jars but one [Re: teladi]
    #25891873 - 03/23/19 10:44 AM (5 years, 9 months ago)

Thanks a lot to both of you for your answers.

I actually went and bought a thermometer to make absolutely sure of the temperature; and it turns out the closet the jars are resting in is at 23.8C (74.8F). So in fact I believe the temperature is close to optimal (I read 24 to 27C is best).

Would being at 24C/75F - the lower bound of optimal temperature - possibly explain such delay in growth ? in any case, I guess I'll just have to wait and pray.

Teladi (is that nickname a X universe reference, by the way :-) ?) : thank you; I'll probably follow the Agar suggestion. When you say "You could re-inoculate now", do you mean I could realistically re-inoculate the same jars using one of my leftover syringes in my still air box ? wouldn't using the same jars increase the risk of failure, as they've been sitting there for two weeks and some undesirable stuff could have had some time to grow ahead of the newly introduced spores (even if I don't see contam) ?

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Offlinepoisoned
untitled
I'm a teapot User Gallery


Registered: 04/17/13
Posts: 1,738
Loc: Yurop
Last seen: 2 years, 2 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25891876 - 03/23/19 10:47 AM (5 years, 9 months ago)

24-27ºC is old information. 24 is perfectly fine and you can go way below that.


--------------------
How I do glass dishes

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OfflineZsuzsi
Stranger
Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25891892 - 03/23/19 10:55 AM (5 years, 9 months ago)

Thank you, so I'm indeed at a loss to understand why almost none of them are growing. Hopefully the spores were just old when they were sent, or I didn't realize it but I injected less close to the jar surface than I thought, and some growth will show up eventually.

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OfflineFun-G
Dr gonzo
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Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25891924 - 03/23/19 11:17 AM (5 years, 9 months ago)

I wouldn't re-inoculate the jars. Best to make up fresh pf cakes.

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Offlineteladi
FUNKSOULBROTHER

Registered: 06/27/17
Posts: 1,189
Loc: South Africa Flag
Last seen: 1 year, 5 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi] * 1
    #25893314 - 03/24/19 07:17 AM (5 years, 9 months ago)

Quote:

Zsuzsi said:

Teladi (is that nickname a X universe reference, by the way :-) ?) : thank you; I'll probably follow the Agar suggestion. When you say "You could re-inoculate now", do you mean I could realistically re-inoculate the same jars using one of my leftover syringes in my still air box ? wouldn't using the same jars increase the risk of failure, as they've been sitting there for two weeks and some undesirable stuff could have had some time to grow ahead of the newly introduced spores (even if I don't see contam) ?




Yep, it is from the X universe. I was obsessed with X3 at the time I created this account.

It might increase the risk. Theres a slight chance that the spores are still germinating. Theres also a slight chance that bad stuff is as well. It's really up to you, 3 choices:

1. Leave it for another 2 weeks. But, spores from syringes are usually quicker to germinate.
2. Sterilize and inoculate.
3  Inoculate again.

Spores sometimes do take a while to wake up.

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OfflineZsuzsi
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Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
Re: 14 days, no mycellium in all jars but one [Re: teladi]
    #25901605 - 03/28/19 08:18 AM (5 years, 9 months ago)

Thank you, I'm not short on jars anyway so I figured I'd just let them sit for another ten days or so (it's now been close to three weeks and still no mycelium, or contam, visible in the 7 inactive jars), and probably start another batch with another syringe in the meantime.

I'm almost tempted to go liquid culture this time but it seems highly prone to contamination so I'll probably follow the same procedure I did initially and hope for the best.

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Offlinepoisoned
untitled
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Registered: 04/17/13
Posts: 1,738
Loc: Yurop
Last seen: 2 years, 2 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25901613 - 03/28/19 08:23 AM (5 years, 9 months ago)

Start with agar first, then proceed to grains and then you'll see if you need LC at all.


--------------------
How I do glass dishes

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OfflineZsuzsi
Stranger
Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
Re: 14 days, no mycellium in all jars but one [Re: poisoned]
    #25901675 - 03/28/19 09:01 AM (5 years, 9 months ago)

You're right, I read a lot more on it and not only does agar seem simpler and less risk-prone, it also seems extremely convenient. I'll have to order petri dishes and the like though; so I'll probably still attempt another PF tek first (and then do agar wedges to brf in a third attempt).

I've taken a few pictures of my jars; both one of the seven uncolonized one and the one that is showing growth. Would anyone knowledgeable kindly tell me if the substrate looks OK, and if the growth itself looks healthy as well ? I'm hoping to point out any mistake I may have done in order to avoid failure during my second attempt.

Here are the pictures:






Thanks in advance !

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Offlinepoisoned
untitled
I'm a teapot User Gallery


Registered: 04/17/13
Posts: 1,738
Loc: Yurop
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Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25901947 - 03/28/19 11:43 AM (5 years, 9 months ago)

Looks really good to me, but wait for someone more experienced to chime in.


--------------------
How I do glass dishes

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OfflineZsuzsi
Stranger
Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25901976 - 03/28/19 12:00 PM (5 years, 9 months ago)

Thanks. I'm especially interested in whether or not the growth medium consistency (verm + water + BRF) appears OK, as I'm at a loss to understand what went wrong.

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OfflineZsuzsi
Stranger
Registered: 03/05/19
Posts: 86
Last seen: 1 year, 6 months
Re: 14 days, no mycellium in all jars but one [Re: Zsuzsi]
    #25952422 - 04/24/19 08:42 AM (5 years, 8 months ago)

Well, an update: it's been more than six weeks and the 7 other jars still haven't shown any sign of mycelium, nor contam for that matter. The good news is the single colonized one seems perfect; it's now dunking.

I've been raking my brain trying to get what I did wrong and I realized I barely shook the syringe before inoculation; my theory is that all the spores clumped together and ended up in the single active jar due to this.

I've already prepped 8 new jars to try again and will make sure to shake the syringe a lot beforehand this time.

I guess at least the single successful cake will allow me to make sure my fruiting chamber is up to par.

Edited by Zsuzsi (04/24/19 08:54 AM)

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Shop: Bridgetown Botanicals CBD Concentrates   PhytoExtractum Buy Bali Kratom Powder   North Spore Bulk Substrate   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Left Coast Kratom Buy Kratom Extract   OlympusMyco.com No Unicorns Here—Just Quality Bags That Work   Kraken Kratom Red Vein Kratom   MagicBag.co All-In-One Bags That Don't Suck   Original Sensible Seeds Bulk Cannabis Seeds   Myyco.com Golden Teacher Liquid Culture For Sale   Mushroom-Hut Liquid Cultures


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