OOOOOOOOoooold spores

[Lono]

Hey mycophiles,

Just wondering what the oldest spore print you ever managed to germinate from was?

I just came across a dung lover print that has been sitting in a cool dry place sealed in plastic for more than three years. I heard dry prints can loaded into a syringe and left for a day or two to rehydrate. But three years is pretty dry right?

Any idea if I'm likely to have success germinating these or should I just keep watching swapnsell?

[PD.]

might as well try it man, what have you got to lose?

It should be fine.

[auto]

Any idea if I'm likely to have success germinating these or should I just keep watching swapnsell?

The oldest success I've seen posted is 2.5+ years, but I agree with just try. It costs nothing but time. Most of the fun is to be had developing your technique and equipment and knowledge and appreciation.

Patience is your strongest ally. If you're too keen, the whole affair will frustrate you, and your mushrooms will suffer it and kick your arse when the time comes.

Try raising anything - tomatoes, flowers, children. The joy isn't the end result, but the sum of the nurturing.

A trip lasts a few hours, but love is forever.

Edited November 22, 2006 by auto

[Hyphal]

Hey man, just dug this up -

http://www.shroomery.org/forums/showflat.p...&PHPSESSID=

I say give it a crack! You should be fine.

[BlackDragon]

Couldnt get that link to work for me(brand new comp this week) so wasnt sure what there but very old spores can be a bugger to germinate but eventually some will.... eventually...

Try making up say halfa dozen or small 200ml jars liquid innoc style. Saftey in numbers! Dont forget to punch or double puch the lids, and tape em up or use the air-port kinda system. This time when making your solution use only a tiny bit of malt extract/honey/karo whatever.. say 1% or bit less, thats all. Sterilise as normal, allow to cool, and innoculate the jars with your spore as normal. Put the jars in a dark cpboard, maybe a few in a warmer place. Ive left jars this way for over 6 months to 1.5 years or more now, and eventually a few spores somewhere in the mix will jump to life. Ive resurected a strain from minimum 3 years old, so im sure its possible. Just he luck of the draw and maiking a few extra jars just helps your chances due to losses thru contamination...

Hope that helps a bit, and hope im not repeating that previous link!

Like the others said, give it a bash!

BD

[Lono]

Cheer's

I think I'm going to try innoculating agar dishes from the syringe and then if anything sprouts make the jars up using mycelium. That way I can see if the prints still viable without having to spend a day sterilising jars and substrate and my kitchen, making poo tea (works great, not as weird as it sounds) etc etc.

I've never cultured mycelium for a jar tek before, does anyone know how to innoculate with it. Do you make a mycelium syringe somehow or just lay it on top of the substrate and cap the jar?

Seems to me, with CO2 being heavier than air, it will pool in the bottom of the jar starving the myc of O2 if it starts growing from the top down.

[gerbil]

There are many ways to achieve the same thing;

1) Streak spores on agar / inject spores on agar (I prefer streaking)

2) Monitor and subculture healthy mycelial tissue to another agar plate; do this as many times as needed until you've got a completely clean, semi colonised agar plate (Best not to let it colonise the entire plate for easier visual inspection and you're aiming to keep it's growth flowing, not stop and starts, but it can still work either way.

3) Prepare your grain jars, let them cool in a glovebox or similar

4) In the glovebox... VERY quickly and as sterile as possible (you'll have to practice to get hand movements flowing), use a scalpel to cut wedges of colonised agar and at the same time transfer your desired amount into the grain jar waiting; to do this you need to have the grain lid loosened and quickly drop an agar wedge and screw lid back on immediately; cutting the agar, transferring and capping the grain jar will be the most likely time for contamination...and frustration lol...Shake the jar (not too violently) to get some even distribution relative to the amount of innoculant.

5) Incubate and monitor, immediately treat and throw out contaminated jars

It's best to create as many agar plates of a healthy culture as possible, that way if you fail on one attempt; you've still got some healthy tissue to be able to work from and don't need to start from scratch again.

In all honesty, spend the money and purchase Stamets texts and/or related texts, they are incredibly valuable and you'll use them for life, it's money well spent.

Edited November 22, 2006 by gerbil

[BlackDragon]

I was actually taking about liquid mycelium culture jars, not grain jars. Both grain jars and agar plates/dishes will/may dry out well before some spore may sprout. Like i said, 1% malt etxract/honey/whatever and just plain tapwater, just use baby jars if you have limited space. If you cant sterilize for 20min in a pressure cooker, or pasturise in a pot of boiling water for an hour.

The spores can take many months to germinate when they do germinate, you can just clean up the now mycelium rich innoculation liquid with some hydrogen peroxide(1ml of 4%/200ml jar will do fine) then you can wait a few days for mycelial recovery and then go ahead and use your fresh and contam-less innoculum to bang up your grain jars.

Had far better sucess with liquid than agar for old spores...

Yep as G said, def get a hold of stamets books if you even half keen on following your new found hobby

Bd

Edited November 23, 2006 by BlackDragon

[occidentalis]

4 years.

[hairyplant]

A trip lasts a few hours, but love is forever.

... that made me smile

nice to see our persuits so sweetly placed into perspective

[gerbil]

apologies BD, I missed your post first time 'round; sounds much more suited to this pursuit, esp. if they are stubborn spores hehe

Can visualise your process description well; makes me want to get back into playing with liquid cultures again.

[auto]

Thanks for peeps input on this. I haven't considered liquid innoculation before.

4 years - thats promising. It looks like a few peeps need (have needed) to germinate old spores. Maybe mushroom cultivation is the kind of project people return to after long periods away from it - like home brewing.

This thread is an excellent resource for the archive. Good work everyone!

[AndyAmine.]

Thats assuming the print is fully sterile otherwsie the LC will contam very quickly, it can quite unforgiving with prints;.

[spiders]

I do everything in agar - everything is a pain in the arse in my experience. Just streak them and wait. i cant see why they wont - might just take longer to come to life.

[Lono]

Thanks for peeps input on this. I haven't considered liquid innoculation before.

4 years - thats promising. It looks like a few peeps need (have needed) to germinate old spores. Maybe mushroom cultivation is the kind of project people return to after long periods away from it - like home brewing.

This thread is an excellent resource for the archive. Good work everyone!

Your dead right auto, especially since my work computer blocks shroomery.org .

[BlackDragon]

Liquid innoc can have a higher rate of contam, hence the numbers game. Contam will usually show well before old spores have germinated(first week), just throw out the contaminated ones asap. When there is signs of mycelium growing, introduce a ml or so of 4% perxoide. The peroxide will clean up any left over contaminants, as well as resupplying a little O2 upon its breakdown, kinda freshening the innoculum.

Bd.

[auto]

Liquid innoc can have a higher rate of contam, hence the numbers game. Contam will usually show well before old spores have germinated(first week), just throw out the contaminated ones asap. When there is signs of mycelium growing, introduce a ml or so of 4% perxoide. The peroxide will clean up any left over contaminants, as well as resupplying a little O2 upon its breakdown, kinda freshening the innoculum.

Bd.

Yep. LC is the definitely the way to go.

A print post marked September 2001 (thats 5+ years), germinated into a cottony mycelial ball (around a spore clump) in 7 days. A 5c piece size daub of honey in a 3/4 full baby jar sterilised at 15psi for 20 mins, allowed to cool and spores tapped into jar from sterile scalpel/print scrape without touching the fluid, then incubated in a double tub w/ aquarium heater kit at 26-28'c range (checked with cheapo Bunnings min/max thermomemter).

The print remained sealed in a ziplock inside double enveleopes at "room" temperature, but was stored in a Perth roof space for a couple of seasons (not exactly ideal temperature conditions). I suspect ambient temps between 2'-40'c are survivable, providing there is no UV or light exposure.

Kudos, BD

[incognito]

dumb question- does the mycelium need air exchange during its growth? could u just prick a hole in the baby food lid and pack with polyfill? it would make things easier and not expose to contams while drawing up in a syringe(through the hole with polyfill) as well for inoculation?

does the 5c dollop of honey in normal tap water sterilised for 2 hrs at 15 psi suffice?

and how do u tell if ur pc is at 15psi if it has no gauge? i just run my cooker at its lowest temp(not hissing lol) after ive brought it to full steam before adding the jars.

liquid innoc looks to be the go,

any links about that spells things out fairly basically for preparing liquid culture?

ive only ever had fleeting success with spore syringes.

have some gr8 prints now so dont wanna fuk it up, got atlantis,tamp AND mexicana to go under the microscope, and dont wanna bodge it!

found this tek... some mycology teks i deadset rekon u need a degree in brain-surgery. this seems pretty straight forward.

http://www.shroomtalk.com/forum/index.php?...p;mode=threaded[/url

seems easy enuff http://www.shroomery.org/forums/showflat.p...&PHPSESSID=

Edited December 20, 2007 by jono

[Aya]

dumb question- does the mycelium need air exchange during its growth?

Indeed it does, with out FAE (fresh air exchange) the mycelium will probably stall, or at best take a long time to fully colonise.

could u just prick a hole in the baby food lid and pack with polyfill? it would make things easier and not expose to contams while drawing up in a syringe(through the hole with polyfill) as well for inoculation?

You could do that, would be a good way of having a sterile environment for the spores while making a syringe.

have some gr8 prints now so dont wanna fuk it up, got atlantis,tamp AND mexicana to go under the microscope, and dont wanna bodge it!

You have an Atlantis, Tampenensis and Mexicana Print?! You lucky man Jono. If you ever happen to come across multiple prints of any, or all of those, and are looking for a trade, Please feel free to PM me. Have been looking for those for what seems a life time... Cant wait to check them out under the Mycroscope!

Edited December 20, 2007 by Aya

[incognito]

if im succesful ill put em bak into z community.

they are out there.

[Aya]

if im succesful ill put em bak into z community.

they are out there.

Well i'm wishing you all the best Jono.

Godspeed!

[Hyphal]

dumb question- does the mycelium need air exchange during its growth? could u just prick a hole in the baby food lid and pack with polyfill? it would make things easier and not expose to contams while drawing up in a syringe(through the hole with polyfill) as well for inoculation?

does the 5c dollop of honey in normal tap water sterilised for 2 hrs at 15 psi suffice?

and how do u tell if ur pc is at 15psi if it has no gauge? i just run my cooker at its lowest temp(not hissing lol) after ive brought it to full steam before adding the jars.

liquid innoc looks to be the go,

any links about that spells things out fairly basically for preparing liquid culture?

ive only ever had fleeting success with spore syringes.

have some gr8 prints now so dont wanna fuk it up, got atlantis,tamp AND mexicana to go under the microscope, and dont wanna bodge it!

found this tek... some mycology teks i deadset rekon u need a degree in brain-surgery. this seems pretty straight forward.

http://www.shroomtalk.com/forum/index.php?...p;mode=threaded[/url

seems easy enuff http://www.shroomery.org/forums/showflat.p...&PHPSESSID=://http://www.shroomtalk.com/forum/ind...&PHPSESSID=

Just to clear some things up mate - you dont want too much honey, around a teaspoon per 250mls is fine - and a 50/50 mix of honey and pure maple syrup I heard works absolute wonders....

Also, you definitely dont want to sterilise your LC for 2 hours! 20-30 mins is ample, otherwise you'll caramelise the sugars.

You know when your PC is at 15 PSI when the rocker on the top will starts rocking back and forth - you want to then adjust the stove temp to keep the rocker doing its thing gently. Be careful man - they can be dangerous, it should have instructions, and if not, research proper pressure cooker use - there is a bit of info HERE.

Also, this guide is supposed to be a relatively good way of making a liquid culture that doesnt use a PC - HERE.

Lastly, colonising mycelium needs gas exchange, not freash air exchange. FAE is for fruiting, gas exchange is for colonising, but both equally as important.

Edited December 20, 2007 by Hyphalknot

[ENtiTY]

dumb question- does the mycelium need air exchange during its growth? could u just prick a hole in the baby food lid and pack with polyfill? it would make things easier and not expose to contams while drawing up in a syringe(through the hole with polyfill) as well for inoculation?

does the 5c dollop of honey in normal tap water sterilised for 2 hrs at 15 psi suffice?

and how do u tell if ur pc is at 15psi if it has no gauge? i just run my cooker at its lowest temp(not hissing lol) after ive brought it to full steam before adding the jars.

liquid innoc looks to be the go,

any links about that spells things out fairly basically for preparing liquid culture?

ive only ever had fleeting success with spore syringes.

have some gr8 prints now so dont wanna fuk it up, got atlantis,tamp AND mexicana to go under the microscope, and dont wanna bodge it!

found this tek... some mycology teks i deadset rekon u need a degree in brain-surgery. this seems pretty straight forward.

<a href="http://www.shroomtalk.com/forum/index.php?showtopic=6025&mode=threaded" target="_blank">http://www.shroomtalk.com/forum/index.php?...p;mode=threaded[/url

seems easy enuff http://www.shroomery.org/forums/showflat.p...&PHPSESSID=://http://www.shroomtalk.com/forum/ind...&PHPSESSID=</a>

Jono your mention of baby food jars leads me to think you are refering to LC for the first part of your post as well. If so, firstly the polyfill would be a very bad idea as if you accidently bump, tip or knock the jar at any point (and you will ) you are going to get the culture medium in the polyfill which will lead to contam. Secondly baby food jars were my first choice as well when it came to playing with LC, in short they are crap. The polymer inside the lid melts when you sterilise in the pc and they never seal properly again. Best idea is to get one of those tall slim olive jars and use that. puch a hole in the lid and seal top and bottom with silicon. I use the silicon designed for use with aquariums. This way you get a self healing hole for your needle. For air exchange in LC I squirt a little H2O2 in when the myc growth starts to slow. Helps with sterility and adds oxygen to the LC at the same time.

Re your PC, its hard to tell. Most of the newer PC's I looked at were only designed to go to about 7psi. You need to check your PC's manual or contact the manufacturer to find out for sure.

For the LC I suggest you use Karo corn syrup. Maybe a bit harder to find but worth it especially if your new to LC. Honey and malt etc all drop proteins out of solution after PCing which makes the LC cloudy and leaves a sediment making it much more difficult to spot contam. Karo comes out of the PC crystal clear so you can see what you have clearly. Like Hypha said 20 min in the PC is all you need for LC sterilisation, any more and you will start to caramelise the sugars which the mycelium tend not to like.

LC seems like the way to go but not worth the effort if you just want to grow some mushies every now and then. LC has its uses don't get me wrong, such as if you were innoculating mass amounts of spawn or breeding a specific high quality strain. If you are having little sucess with spore syringes then you are going to have trouble with LC. You still need to make a spore syringe to inocc the LC . Using LC adds another step which adds another possibility for contam to crash the party. If this were me I'd perfect the basics before getting more adventerous.

WOW! Atlantis, Tamp and Mexicana eh. I know some members that have been keen to get them under their microscope since before I signed up here. Half your luck

Gluck Jono!

Edited December 20, 2007 by Harry

[Hyphal]

For air exchange in LC I squirt a little H2O2 in when the myc growth starts to slow. Helps with sterility and adds oxygen to the LC at the same time.

LC seems like the way to go but not worth the effort if you just want to grow some mushies every now and then. LC has its uses don't get me wrong, such as if you were innoculating mass amounts of spawn or breeding a specific high quality strain. If you are having little sucess with spore syringes then you are going to have trouble with LC. You still need to make a spore syringe to inocc the LC . Using LC adds another step which adds another possibility for contam to crash the party. If this were me I'd perfect the basics before getting more adventerous.

Harry, all really good advice man except for those points - there's definitely no need for H2o2, this has shown to do more harm than good... If you want to introduce air to your LC jar, just make a second hole and do the silicone thing again. Then insert a syringe that has the plunger removed and the body stuffed with poly fibre wool to act as a filter in through the second hole. As you suck up Liquid through the first hole, sterile air gets sucked in through the other other poly-stuffed syringe.

Also, you are wrong about needing to make a syringe - you can innoculate straight into your LC by cracking the lid and dropping some spores in off a sterile knife tip - a glove box is handy here, or a very clean kitchen bench, clean gloved hands and a still air environment (shut windows and doors etc.).

IMO, LC should be an essential step in the growing process. Once scrape of spores can give you 250ml of LC ready to use in 2 weeks, and that will last in a dark cupboard for up to 12 months or even longer in the fridge. Whenever you need to, simply suck up a few mls and your good to go.

Jono, cover all bases and use 3/4 of one print for a syringe and the last of the print to make an LC.

[ENtiTY]

Harry, all really good advice man except for those points - there's definitely no need for H2o2, this has shown to do more harm than good... If you want to introduce air to your LC jar, just make a second hole and do the silicone thing again. Then insert a syringe that has the plunger removed and the body stuffed with poly fibre wool to act as a filter in through the second hole. As you suck up Liquid through the first hole, sterile air gets sucked in through the other other poly-stuffed syringe.

Also, you are wrong about needing to make a syringe - you can innoculate straight into your LC by cracking the lid and dropping some spores in off a sterile knife tip - a glove box is handy here, or a very clean kitchen bench, clean gloved hands and a still air environment (shut windows and doors etc.).

IMO, LC should be an essential step in the growing process. Once scrape of spores can give you 250ml of LC ready to use in 2 weeks, and that will last in a dark cupboard for up to 12 months or even longer in the fridge. Whenever you need to, simply suck up a few mls and your good to go.

Jono, cover all bases and use 3/4 of one print for a syringe and the last of the print to make an LC.

H2O2 do more harm than good? Where has this been shown and what kind of harm? H2O2 is simple and involves very little risk of introducing contam. The air sucked in through a syringe stuffed with polyfill is not even close to sterile. You would need at least a .3 micron filter to block all the nasties via that method.

Yeah I know there are other ways of knocking up LC, its just when you compare the simplicity of making a spore syringe against the cumbersome and awkward glove box, there isn't really even a choice. Keeping things as simple as possible makes for a more sucessfull experience I find. I understand the logic behind the use of LC but unless you want to produce heaps of spawn it is a waste of time. In the 2 weeks it took to grow the LC those same spores could have colonised a grain jar and be ready to case or spawn to something. Sure the LC route may give you a couple of days head start the next time around but most of us aren't in a commercial situation where time is money, we are just hobbiests and a couple of days doesn't matter that much. Anyway if your like me the next grow will be a different variety and a print is much more easy to store than LC and will last much longer as well.

I'm not trying to be picky here but 3/4 of a print seems a bit wasteful. Depends on how dark the print is but even on the lightest print I have had I only use 1/4 of it. Its always good management to save some spores to go back to if something goes wrong.

But yeah each to his own, try both Jono and see which suits your methodology better I guess.