What's my next mycelium move? Get Out of Jail Tek

Started by PCCP, March 26, 2011, 12:37:28 AM

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PCCP

Hi everyone, complete noob here.

Have read and re read the PF-Teks and understand fully what I should of done but I had a major error in judgement.

I've inoculated 6 brf jars with 1.3cc of spores each and put them in my incubator that normaly gets to 28C max during the night, unheated through the day. My place is average 15C so extra heat is needed

I put them to bed and thought I would add a layer of tin foil to keep in complete darkness. I then had to leave.

When I got back 5 hours later the air temp was nightmare 35C. I didnt realise the foil would retain so much extra, as 7C deg. I opened and cooled of straight away and only closed for the night when it had dropped to 25C. This morning was down to 18C

I'm not sure how long the heat was at 35C testing the heater I think it was about 2 hours.

Is 2 hours at 35C too much for freshly inoculated jars?

Will they germinate?

If they germinate are they doomed for less than full colonization due to moisture loss in the early over heated hours?

I should wait to see at least  germination and keep the temp low say 20-22C and humid to prevent any more moisture loss?

Sorry for the long post, just wanted to give more experienced people all the facts.

Peace :huh:


PCCP

#1
I think I may have answered my questions here

http://www.shroomery.org/5159/What-is-too-hot-for-incubation-and-what-happens-to-the-jars

My spores are cubensis.


I was half way to thermal death and slowed any germinated growth down to 50%. These are ungerminated jars so may have got away with it but the substrate may have lost too much moisture in that crucial hot 2 hours.

I think I'll wait it out and keep my growth temp close to room temp as I dare to keep any more moisture loss to an absolute min.

:ph34r:

PCCP

No signs of white yet.

I'm using a timer with my incubator. Can't afford the thermostat just yet.

I've manage to stabilize my temp in the 22-26C range, difficult at night as my ambient temp plummets, but I think I can refine the temp band a little more with a few more tweaks.

I've been reading it's possible to salvage a less than fully colonized cake by cutting off uncolonized substrate, then crumble 'n' case.

Is this worth thinking about on my first grow since I only got 2cc of spores left. I wanna spend next months extra cash on a reptile hydro-thermometer, not more spores.

Any input is cool on how to manage my fuck up. :rolleyes:

Cheers Everyone :ph34r:

Vandal_xx

Dude, hang in there. Soon you can make your own spores
Feel good before dying!!!!!

PCCP

Cheers Vandal,

I don't see white yet but I ain't giving up. :mellow:

PCCP

Still no good stuff, but I got no bad stuff either.

I'm beginning to question the spores themselves.

I might test them on a single Liquid Culture Jar

:mellow:

PCCP

After 11 days I have white in 1 of 24 inoc points YEEEES.

Come on you whites

:ph34r:

Vandal_xx

I'm thinking that the syringe you got had very few spores in it. I made a syringe out of a very light print and now my jars are barly showing any white. Hopefully you will get some fruits so you can makeyour own highly sporilated syringes. good luck
Feel good before dying!!!!!

PCCP

As I said in my previous post I was beginning to question the quality of the syringe it self.

It came from a mycology and mushroom company here in the UK. Due to questionable legality of selling spores they ask not to be contacted with regards to growing.

This made me think in retrospect that they could easily supply a very weak syringe and not have any comeback.  :huh:

If I do need more spores I won't be returning for there services.  :angry:

It was a Colombian strain.
It looked clear with slight tone.
I created an air bubble and then shook before each inoculation.

If I can get some LC going from 1 of the 6 Jar then that would be great.

I may use the last of the 2cc of spores to try LC from that angle too.

Thanks for the feed back Vandal

:mellow:

PCCP

So 2 more days and it will be 3 weeks since inoc.

I have 1 out of 24 inoc sites that germinated . Its produced think white myc that has shrunk away from the jar and spread the size of a golf ball.

The bottom of the jars has dried due to the initial over heating.

This myc is very precious as its the only bit I managed to develope and bring this far.

I seriously dont think the jar will achieve 100% colonization.

What can I do from hear? :

1) Make the myc into LC
2) Break of all uncolonized brf and try and grow a single shroom from there.
3) Add sterile water to the cake with a syringe

Please help, any suggestions  :huh: ^_^

-_-

PCCP

#10
24 days since inoculation.

4 inoc points in each jar.
1 out of 24 ionc points germinated.
This 1 point in my only successful jar has grown from the previous golf ball to 8 cm. This is more than 33% of the Jar. Very thick white.
The bottom of the Jar seems too dry to colonize.

My plan is to inject this jar with 20cc of sterile water, shake vigorously, then withdraw 10cc or what I can to start a LC.

With this idea I hope to hydrate my jar and at the same time continue the propagation of my only myc.

Is this a good injectable amount?
Will withdrawing fluid back impede any further colonization?
Should I inject only and leave to colonize more?

Any thoughts?

:huh: :mellow:

Vandal_xx

I have no idea what you should do. You can try the LC tek, and if it fails you will be no worse off. Go for it.
Feel good before dying!!!!!

PCCP

It's day 25 since inoculation but germination didn't happen for 11 days.

What I have is 14 days of myc. growth. I think I can push it a little further but I need to add moisture soon or it may just eat the nutrients and leave nothing for any potential flush.

If I add water to it to help hydration I may also be introducing contams but it could be the thing that helps it to 100% colonization. I may also get an LC to continue propagation.

I need some experienced help on this.

???

Hanging in there... :ph34r:

PCCP

Day 30 since inoculation.

19 days of myc growth after 11 day germination.

11.5cm size across, v. very thick dense white, shrunk away from glass, 25-27C

Didn't think I would get much more growth but I think its possible now. There are still moist areas to colonize but the dry parts are becoming loose and clearly visible mainly the 5mm base.

I'm going to attempt an LC extraction with the idea it will help lost water back into the Jar also with what ever moisture is left behind.

Vandal_xx

You can also let it grow as long as it keeps growing and then cut/scrape off the uncolonized parts and put the colonized peice in the FC. You may have some luck.
Feel good before dying!!!!!

PCCP

Quote from: PCCP on April 13, 2011, 01:50:43 AM

2) Break of all uncolonized brf and try and grow a single shroom from there.


Cheers Vandal,
This option has always been in my armory, but as I wait and try to be patient to see what the best move is gonna be as the myc is still growing this could be the one

It's still growing so I'm still hanging, waiting and thinkin'  :ph34r:

PCCP

Day 45 since inoculation.

34 days of myc growth after 11 day germination.

Fully colonized the main body with v. very thick dense white, shrunk away from glass, only the bottom 5mm that's dry has failed. 25-27C

I'm stocked. Didn't think I would get this far. I've inverted the jar to help clear any CO2 while I prepare my LC jars over a couple of days.

I'm going to attempt an LC extraction with the idea it will help lost water back into the Jar also with what ever moisture is left behind.

I think after the hot start and delayed germination I may have some heat tolerant mycelium on my hands.

  :mellow:

PCCP

Day 47 since inoculation.

36 days of myc growth after 11 day germination.


I used my last 2cc of spores on 2X LC medium (malt dextrose 4% solution) 1cc each. Glass in bottom for agitation and each jar half full.

Injected 30cc malt dextrose 4% solution – extracted 5cc for a 3rd seperate LC from the only cake. It was left pretty wet and bruised.

I figured 3 LC's was a better risk than 1.

It was so dry it soaked the lot up. I really struggled getting any back out.
I wouldn't recomend this tek if your cake is really dry as I had to put more in than I wanted so it would give some up on return.

Everything returned to incubate for a few days, cake to recover etc. Agitating LC daily.

A stressful success so far.

:ph34r:






BabyBlue

Good luck man. I would buy some new spores and get going on another batch. Maybe you shouldn't try so hard to regulate temperature; I just let mine sit in an enclosed sterile box. It's possible the heat could be hotter than you think, and could be negatively effecting germination. But this is simply speculation, since I'm only 2 days into my first grow.  :rolleyes:

Best of luck!

PCCP

Cheers BabyBlue,
You may not of read my first post. I basically cooked the spores by mistake for a couple of hours. I won't fall into that trap again. One learns from mistakes  :rolleyes:

I believe the spore syringe I had was very thin any way but still got 1 out of 6 to germinate. The full colonization was from 1 of 4 inoc sites so I think 36 days aint too bad. If all 4 inoc points were good then it could of been done in 9 days + germination time. I'll aim for 14 days next time round.

The LC tek was to safe guard propagation in case fruiting fails as I can't afford spores every failure.

My temp is now well in the comfort zone (26C tops) but is still essential as my night temp naturally drops to 8C (I live on a boat so insulation isn't house standard)

Thanks for your feed back.

:ph34r: