is there a way to rescue my "colony" ?

[junk1]

Hello there Mycotopians .
This is one of my first posts, as i recently came to idea of my own "champignons". Like ten days ago a friend of mine was coming back from holland and brought me a box of fresh champignons. It was early at the morning, or better late at night so i put it on the fridge.
The very next day, searching some info about how could they "work" etc. i spotted some easy LC, and cloning teks. As it was sunday, my only way was to buy some honey, distilled water and make Honey LC. I did six jars of LC, and put fresh tissues into them, from inside the steam, as courious about sterility as i could. Well as far as now, all jars are contaminated, due to big flow in my thinking... I sterilised all of the shit except... distilled water as i thought it should be allready sterille. Well, there's the situation. In three jars i can see some mycelium grown around tissue bits, they start to grow after like 3 days, but then stalled and LC become a bit cloudy, so i suppose it's contamination . Is there a way to "rescue" those ? I also preserved some dried shrooms "in a case". They were far to small to get spore prints so i got what i could. Well as i dig into shrooms cultivation, i find it quite a fascinating hobby But there's virtually my last chance to revive those. Thanks in advance, for any help.
Best regards

[limeade]

The bits with mycelium growign on them need to be transered to agar. Then transered across at least 10 plates, you'll want to be mindful to transfer away from any contaminates you may have, while selecting the most rhizomorphic mycelium possible.
Your cloudiness may either be just from the honey, which honey usually does this, or it's yeast, possibly even bacteria, all 3 things make LC's go cloudy.

You can do simple agar
or you can use petri dishes with antibiotic agar.

I would say, use a simple agar tek utilizing 1/2pints as "petri dishes" and PDA as your grow medium, potato dextrose agar, u can even peroxidate it at 3% when the agar reaches 135*F, just before solidifying.

hope this gives you more information to search on, the archives have this information.

champignons aka Agaricus Bisporous, grow much like psilocybe cubensis do, agaricus are SUPER picky about their casing layer though, you will want to use a peat based casing layer, I have used a 5 inch manure/straw substrate in the past also with great results.

When i lived in a favorable climate, i made these beds right along side my garden. I don't live in a favorable environment now.

Your dried tissue can be reanimated also!

dry tissue reanimation:

Here's a process that has been 100% successful in reviving dry mushroom tissue for me so far. Don't use peroxide, whatever you do. It's toxic to fungi cells, and the last thing you want to do when reviving tissue is harm it further.

Sterilize several jars of distilled water, one of which should be prepared with small air exchange holes in the lid and a method of filtering, such as tyvek or synthetic filter disk, polyfill, etc.

Break up the dry tissue into grain sized chunks and put a dozen or so into one of your sterilized jars of water. Shake well to wash the pieces of tissue, and then transfer them to the next jar of sterile water. (you can pick up the pieces with a syringe needle or scalpel) Shake again to wash and repeat.

When you place them into the final jar, which is filtered, swirl them around in the water and then set the jar away and forget about it for a few weeks. You'll begin to see mycelium growing on any chunks that are viable. Don't use nutrients of any kind, just plain sterile distilled water. Once mycelium begins to form, transfer to agar to isolate away from any contaminants that may be present.

[junk1]

Thank You very much for your input, i just ordered some agar but i am waiting for order to come. Should i perform some "quick action" to save still living mycelium, or just wait a few days more for agar to come ? I've heard of bleach, in 1:200 measurements. If so, how many cc's should i drop into jars ?

Best regards

[limeade]

I wouldn't suggest the bleach, i think what you have in there will be ok, make a pint of sterile distilled water to wash it in, then pluck it out and put on agar, do all of this after the plate is poured.

if you don't wash the mushroom material off, whatever is growing in your LC with the shroom, will surely take over the agar first,

the quoted section i posted, you can start doing that, so that if your first attempt fails, your tissue will be reanimated in 4 weeks.

If you have enough to pour 20 dishes, i'd suggest only pouring 10, and saving the other 10 for your second attempt after reanimating the tissue.

[junk1]

Thanks to limeade advices, i apparently rescued myc.

Transfered to "agar substitute" or simply thin layer of brown rice flour in a small jar, then inoculated two jars. Effect on the pic This is six days after inoculation, 24 hrs after first shake. Contamination was leftover within one transfer. Next six jars are waiting for G2G but i am quite sure it will work this time! Thanks again Limeade!

[junk1]

Hello again, a few of the jars processed as stated above look like this one:
Do You think it is ready for spawn, or should i wait some more to get myc thicker? it's been 17 days since innoculation, i am planning to rez-effect this in fahtster type monotube, with verm casing. Any advices?

[sagiboy]

i would love to see those "champignons" growing.... lol.....
if those would be contaminated, (and they won`t), ill help ya with some "champignons" ... lol.....

[junk1]

Originally Posted by sagiboy

i would to see those "champignons" growing.... lol.....
if those would be contaminated, (and they won`t), ill help ya with some "champignons" ... lol.....

Thank You for proposal i'll post results for sure.

[crazy1]

yup that is ready to be used as spawn.
Be sure to break the myc up the day before you transfer it though.
This gives it a chance to recover before spawning.

[junk1]

By the way, that's how looks the small jar i used to transfer, quite a rhizomorfic myc i guess . Is it safe to assume, that if i cloned tissue from commercial grown shroom i shouldn't need to isolate as it should be allready best fruiting ?

[junk1]

Hello again, i guess i am too lazy to get it growing so i just tossed all my agaricus mycelium :P Anyway, strange coincidence, some of my FOAOnlineF
asked me some question and from now on i'll be quoting his phrases

"Well since it's my very first grow, i need some advice. This tray is corn with verm (rez-effect) not cased, incubated for 3 days, then put into FC, where it sits for 5 days. It's been misted once a day, but now i am seeing a lot of little white balls , as on picture wchich i am suspecting of being early forming knots,

The greenish tint in the center isn't really greenish, its my camera's flash
Should i stop misting for some days or should i just hold on my imagination ?
Does it whatsoever look allright ?
When to late case this ? "
regards

[junk1]

Anyone?

[Beastmaster]

don't stop now, if its starting to pin then continue with your schedule, maybe decrease misting just a bit and increase the fanning or air exchange rate if you can.

[junk1]

My FOAF sent me an update:
"
it's pinning baby ! after 7 days in FC i've seen this at the morning:

Counted eight of them till now and a loads of knots. Well since those are my first pins ever ... .
Thanks to everybody that cared .
"

Originally Posted by Beastmaster

don't stop now, if its starting to pin then continue with your schedule, maybe decrease misting just a bit and increase the fanning or air exchange rate if you can.

Since this is standing near vent shaft, it has good FAE i guess, my only concern is watering, since it dries really fast.

[crazy1]

How are you keeping the RH up?
If you're using perilite then just keep adding water as you see fit.
And you could also keep misting the sides of the FC

Lookin good

[limeade]

Originally Posted by limeade

I wouldn't suggest the bleach, i think what you have in there will be ok, make a pint of sterile distilled water to wash it in, then pluck it out and put on agar, do all of this after the plate is poured.

if you don't wash the mushroom material off, whatever is growing in your LC with the shroom, will surely take over the agar first,

the quoted section i posted, you can start doing that, so that if your first attempt fails, your tissue will be reanimated in 4 weeks.

If you have enough to pour 20 dishes, i'd suggest only pouring 10, and saving the other 10 for your second attempt after reanimating the tissue.

WOW!!!

NICE save!

I apologize for not responding i really thought i subscribed to this thread!
I'm very very happy that worked.

If you found that "mini-petri" to be easy, you can go further with transering the more desirable looking mycelium, or even clone with it. I really strongly urge 5 or more transfer so that you get about 4 "petri" to actually work with, make "slants" for long term storage, and have plenty to cut wedges for bad ass grain to grain!

very good work man.

Originally Posted by junk1

By the way, that's how looks the small jar i used to transfer, quite a rhizomorfic myc i guess . Is it safe to assume, that if i cloned tissue from commercial grown shroom i shouldn't need to isolate as it should be allready best fruiting ?

that looks pretty freekin rhizo to me.

I would absolutely hope so! Each commercial grower should be taking the best steps possible to maximize their yield and i'm sure they do, 4 or 5 transers wouldn't hurt to get away from any bacteria that may be present though, at the same time, collecting from the utmost rhizomorphic looking "tendrils" of mycelium present. You'll be sure to have a commercial grade winner at that point.

yesterday you saw pins how they lookin now?

Oh, and your colonized jar pic, i've noticed that when your myc looks somewhat "powdery" like that, it's pretty aggressive.

[junk1]

Originally Posted by crazy1

How are you keeping the RH up?
If you're using perilite then just keep adding water as you see fit.
And you could also keep misting the sides of the FC

Lookin good

"
Well this is my "setup" :

It's just cheap washing powder box from any supermarket.
Holes covered with 3M micropore tape i were misting its wall and slightly surface, till first knots, now i am just misting walls.
I've got two similar boxes, one is standing near vent :

And the other just on some shelf. They are both watered equally, but the one near vent is FAAAAR ahead the other. I guess finding some draught area is good advice. Both boxes contain same (pint / half liter) jars of corn mixed with equal amount of moist Vermiculite. The one on the shelf doesn't even show a sign of knots, while the other is pinning everywhere.

Originally Posted by limeade

WOW!!!

NICE save!

I apologize for not responding i really thought i subscribed to this thread!
I'm very very happy that worked.

If you found that "mini-petri" to be easy, you can go further with transering the more desirable looking mycelium, or even clone with it. I really strongly urge 5 or more transfer so that you get about 4 "petri" to actually work with, make "slants" for long term storage, and have plenty to cut wedges for bad ass grain to grain!

very good work man.

Thanks for appreciating my efforts

Originally Posted by limeade

that looks pretty freekin rhizo to me.

I would absolutely hope so! Each commercial grower should be taking the best steps possible to maximize their yield and i'm sure they do, 4 or 5 transers wouldn't hurt to get away from any bacteria that may be present though, at the same time, collecting from the utmost rhizomorphic looking "tendrils" of mycelium present. You'll be sure to have a commercial grade winner at that point.

yesterday you saw pins how they lookin now?

Oh, and your colonized jar pic, i've noticed that when your myc looks somewhat "powdery" like that, it's pretty aggressive.

I live in europe, so that's a bit different time zone (six hours)
Here's developing on 24 hours time span:
Yesterday's pic again:


Today(24 hrs later):


There's more and more of them every few hours if they won't abort, i'll be quite surprised since i didn't even dream about that kind of pinning

And well, i did as You said those are only two jars, from first transfer,
there are more to come, at various stages of colonization.
At first my "agar plate" wchich i consider just "agar substitute" as on shroomery ( brf with some water in small jars) went sour smelling and kinda yellowish. I just took some myc that developed as well with small sterilised pliers, and moved onto new two plates. Both were clear myc after a few days. Then i put some bits of it into two jars, the next were transfered 4 times more, tried to get as "ropy" myc as i could. Well i hope that bin wouldn't dissapoint me and it's Thanks to You mycotopians! .

And.. well sorry for that much blah blah and poor english.

"

[limeade]

looking great!~


I cannot urge one to fan enough , fan fan fan.

[TastyBeverage]

Your english is great! Very nice grow, looking forward to more pictures.

[limeade]

hahah thought those "champignons" looked familiar, nice "agaricus"

[junk1]

Originally Posted by limeade

hahah thought those "champignons" looked familiar, nice "agaricus"

Well i guess it must be Agaricus silvaticus since it has those strange brown caps

[ernestro]

looks very nice! Fan as much as you can and you may get a supadupa pinset, looks promising

[junk1]

"Well it's nothing much to talk now so that's today's update, whole bin this time:


It kinda slowed down, since temperatures at my apartment dropped to like 65 deg. Counted like 70 pins, hope it's maybe not great, but just OK as for first grow . Guess it became kinda grow log or sth Every pic was taken at the day it's been posted.

[limeade]

lookin bad ass mate!