Author Topic: Mushroom surface sterilization  (Read 12555 times)

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Yachaj

  • Guest
Mycro is the way to go, Bujinkan
« Reply #20 on: September 01, 2002, 04:35:00 AM »
Potency does not decline as a result of more nutrients.

Potency declines as a result of storage. Which is necessary if you do not use the harvest immediately. Bulksubstrate means 'a lot of substrate' and a big harvest.

A pressure cooker sterilizes more thoroughly.

Not important. 'Effective' is good enough for me. Especially if it can be achieved by a more simple approach.

bulksubstrates show an alkaloid content of 0.7 at best.
as compared to what? cakes?

Example assays:

Cubensis on bulksubstrate, potency 0.5%

http://diseyes.lycaeum.org/teo/cntc.txt



Cubensis in the wild:
see Stamets, PSILOCYBIN MUSHROOMS OF THE WORLD (I do not have it by hand right now but it gives a list of assays, none of which comes above 0.7 percent)

Cubensis on rye:

http://jeremybigwood.net/JBsPUBS/JBScientific/VariationOfPsi/pages/Variation3.htm



Cubensis on brown rice:

Jochen Gartz, patent No. 88-09773, Akad. Wiss. DDR, noted over 1 percent alkaloid content. If I find back the abstract I will post it.

-daily maintanance is required with any shroom setup except sclerotia and invitro.

Exactly! Invitro ('mycro tek'] is the way to go. Finally a way to cultivate mushrooms on the road in your backpack! (tested last year while traveling through Europe - works wonders).

-not everyone wants shrooms for 'experiments'

I know. Some people have still not drawn any conclusions from Spitball, Strike etc. And of course there are always some DEA members of the HIVE who are very interested in larger scale methods and the people who use those. It is just not my cup of tea. I am of the opinion that illegal substances need to be destroyed in some way as soon as they are indentified as such. No distribution - no storage.

all techniques that ive ever heard of are contained here, for free. all you need to do is read all this stuff.

http://www.shroomery.com/findorgrowthem.php%5B/blue

]

That is the problem exactly ('read ALL this stuff'). It is too long and useful methods are mixed up with not useful ones. The manuals I mentioned contain just what you need to know to be successfull.

this i am baffled by...who told you this? most of the time the growing surface is completely covered with mycelium by the time pinning starts...and even if it isnt, by the time the mature mushroom drops spores and those germinate it will be totally overtaken with mycelium.


The germination of the new spores on top of mature mushroom caps happens in the stages as depicted on the first and the last picture you posted. In the first it is in the blue tray at the left. Spores often also germinate on the gills.

I dont know where you get your info from, but you need to check your sources..seriously.

I get my information of careful observation, followed by experimentation.

Yachaj

bibliopharmacophile

bujinkan

  • Guest
myc
« Reply #21 on: September 01, 2002, 05:08:00 AM »
The germination of the new spores on top of mature mushroom caps happens in the stages as depicted on the first and the last picture you posted. In the first it is in the blue tray at the left. Spores often also germinate on the gills
I think you are confused about terminology or something....the first and last pics show open caps depositing spores on other caps under them. however, the spores there are not germinating...why would they? And the last place the spores would germinate is in the gills. this is where spores are created by the organism to be released into nature to find other nutrients to colonize.

Potency declines as a result of storage. Which is necessary if you do not use the harvest immediately. Bulksubstrate means 'a lot of substrate' and a big harvest.
Properly dried shrooms can last for months (and usually longer) without losing much potency. as long as you dont dry them with heat they lose little in the way of alkaloids and stay that way for a long time.
more to come later.

If I'm reading this correctly, and I like to think that I am.....

Yachaj

  • Guest
Applications of fungal autocannibalism
« Reply #22 on: September 01, 2002, 06:21:00 AM »
the spores there are not germinating...why would they? And the last place the spores would germinate is in the gills. this is where spores are created by the organism to be released into nature to find other nutrients to colonize.

Fungal biomass actually is the best possible substrate for spore germination. Fungi are superb autorecyclers (it is the reason that yeast extracts as Marmite (tm) work so well as agar additive). Fungi use this quality to grow new organs by using their old organs as food. With bacteria, fungi share the feature that death is not a part of their normal life cycle. A part of the organism produces new spores, which often germinate on older parts of the mycelial mat (or mushrooms), which then are recycled. Spores have a much better chance for survival when they germinate on the mushrooms of their own species compared by a lonely germination on a cow pie in the midst of millions of contaminants.

If spores fall on the mycelial mat it is also possible that the hyphae which sprout from the spores 'invade' the mycelial network and take it over.

See for a full explanation of all the evolutionary advantages of fungal selfrecycling and genetic takeover strategies for instance the book of the British mycologist Alan Rayner:

Alan D. M. Rayner - DEGREES OF FREEDOM, Living in Dynamic Boundaries (especially chapter 5 p. 155-179)

The far majority of the germinated spores of cubensis and Pleurotus come to life on the gills.

Properly dried shrooms can last for months (and usually longer) without losing much potency.

Exactly. They loose some potency and are a legal hazard. Nothing beats fresh harvested mushrooms. Mushrooms which are not harvested and not grown for drugs but for instance to to study mushrooms or as ornamental are not illegal.

A good overview of reasons to choose Psilocybe cubensis as model-organism to study the growth of agaricales is in:

Edmond R. Badham - CULTURAL STUDIES ON THE MUSHROOM PSILOCYBE CUBENSIS (Ph.D. dissertation 1983, University Microfilms, Ann Arbor, Michigan)  

bibliopharmacophile

bujinkan

  • Guest
Exactly! Invitro ('mycro tek'] is the way to go.
« Reply #23 on: September 01, 2002, 12:17:00 PM »
Exactly! Invitro ('mycro tek'] is the way to go. Finally a way to cultivate mushrooms on the road in your backpack! (tested last year while traveling through Europe - works wonders).
if thats what your needs or wants are then invitro is fine.
Not important. 'Effective' is good enough for me. Especially if it can be achieved by a more simple approach.

It is important. it reduces the likelyhood of contamination considerably. And with certain substrates its essential...like when you want to use birdeed for spawn.

as for your links on alkaloid content, all i see is two studies showing alkaloid contents at about .7...where is the average alk. content for mycro tek? all you allude to is cubes on brown rice, which were probably grown in a terrarium on rice cakes. Either i just dont get your point or its some kind of smokescreen.
I am of the opinion that illegal substances need to be destroyed in some way as soon as they are indentified as such. No distribution - no storage
my local DEA office feels the same way. :)

Spores have a much better chance for survival when they germinate on the mushrooms of their own species compared by a lonely germination on a cow pie in the midst of millions of contaminants
this paragraph can be interpreted many different ways...how old is the tissue that the spores are geminating on? how much, if any does this lend to the strength of the organism? logic dictates that the amount of building materials is the variable upon which the size, potency, and longevity of the organism depends the most..
and if you try to let shrooms rot in an indoor setup in order to let the mushroom recycle itself, youre going to have a contaminated mess, especially if you dont wash your hands.
and furthermore, what does any of this matter if you are doing an invitro tek? theres going to be very little spore dropping invitro.



If I'm reading this correctly, and I like to think that I am.....

paranoid

  • Guest
a welcome clarification
« Reply #24 on: September 02, 2002, 09:40:00 AM »
"I see and agree that I should have formulated my statement better. I should have written that a mycro (or should I write 'nano' here?) sterile environment is sufficient. You do not need a sterile environment which is big enough to live in (a sterile room), nor do you need a sterile environment which is big enough to stick your arms/hands in (glove box, flowhood). An environment which you can hold in your hands while standing in the kitchen is good enough."

This clarifies your intent immensely.  My interpretation was that you were implying a sterile environment wasn't necessary period.  My apologies for jumping on you so quickly, but it seemed that you were either completely out to lunch or about to launch into an advertisement for your own shroom grow setup.
::)

Yachaj

  • Guest
Short answers
« Reply #25 on: September 04, 2002, 03:18:00 AM »
To paranoid: I understand. But do not worry I am not about to sell anything. Not on this site. About nano-sterile environments - the difference between no sterile environment and a miniature sterile environment evaporates at the moment that the size of the sterile environment is not much bigger than the cell wall of the spore/hyphe itself. That is what happens in nature and I am convinced that it will created artificially eventually. We just need to look&think deeper.

To bujinkan: You are right, I have no actual figures of the potency of cubensis on 'PF Substrate'. But PF Substrate is just powdered brown rice, vermiculite and water. I think it is OK to compare this to just brown rice, which resulted in cubensis of a potency of 1 percent in Gartz' publications.

with certain substrates [a pc] is essential...like when you want to use birdeed for spawn

Not if you mix the birdseed with something which makes the substrate more airy and use containers which do not exceed 250ml. But who needs birdseed?.

how old is the tissue that the spores are geminating on?

Usually the spores germinate on over-ripe mushrooms which are still alive enough to defend themselves (and the spores) against contaminants. The tops of the mushroom caps, where the spores land&germinate, are for the spores what egg yolk is for the embryo of an animal.

Wind&rain break off the over-ripe mushroom and bring it to new substrates - in this way the old mushroom cap becomes a platform from which new mycelium can grow out. BTW - one of the first underground cubensis cultivation techniques, titled FIELD GUIDE TO THE PSILOCYBIN MUSHROOM (1972), promoted the use of cubensis caps as spawn for compost outdoors!

To answer your question more completely I have done a little test this week with 5+ years old cubie tissue. Spores have no difficulty using it as substrate. Old mushrooms are a primo substrate ingredient. I have no figures yet of a PF-like substrate of just water, vermicuklite and cubensis powder but you made me curious. I will look into that!

if you try to let shrooms rot in an indoor setup in order to let the mushroom recycle itself, youre going to have a contaminated mess, especially if you dont wash your hands.

I think you misunderstand me. I do not promote to use the same substrate more than once. But I do advocate to allow spores to germinate on gills and over-ripe mushrooms, then transfer it to 'R. Wayne' substrates and cultivate without PC/hepa blower etc. But that is for oysters and the like, not cubensis. For cubensis the invitro PF TEK is easier.

And you will probably be surprised to learn that in 'invitro' setups, the mycelium often keeps recycling itself (&produce mushrooms!) until the momen that little more than just vermiculite remains.

and furthermore, what does any of this matter if you are doing an invitro tek? theres going to be very little spore dropping invitro.

On the contrary. But for a decent print you need to give the mushrooms enough headspace. A favorite approach of me is to use a longdrink glass as cultivation container, with a double thick contaminant barrier. When the substrate is colonized, most of the layer is decanted and the glass put upside down. Now there is plenty of space in the glass to allow a mushroom to mature and sporulate.

Yachaj

bibliopharmacophile

GOD

  • Guest
Not if you mix the birdseed with something which ...
« Reply #26 on: September 04, 2002, 04:09:00 AM »
Not if you mix the birdseed with something which makes the substrate more airy and use containers which do not exceed 250ml. But who needs birdseed?.

someone who is looking to maximise yeilds beecause birdseed is WAY more nutrient dense, this equals more flushes and stronger shrooms.

with certain substrates [a pc] is essential...like when you want to use birdeed for spawn

Not if you mix the birdseed with something which makes the substrate more airy and use containers which do not exceed 250ml


You have obviously not worked with birdseed.  Although you have done alot of research, nothing beats actual experiance.  Open your mind, and give it a try, you might find that you can formulate an even better plan.

the difference between no sterile environment and a miniature sterile environment evaporates at the moment that the size of the sterile environment is not much bigger than the cell wall of the spore/hyphe itself. That is what happens in nature and I am convinced that it will created artificially eventually.

What exactly happens in nature?  I dont understand what your trying to say, but there are way too many environmental factors in nature to reproduce in an artificial setting, too many organisms, systems etc.. that come into play out in the wild.  Chaos theory applies here.  Indoor cultivation needs sterile environment because indoors breed way way too many competitors in such an enclosed environment.

Go ahead and give your method a try.  Please dont misunderstand whats happening here, there appears to bee a few among us who have a fair amount of experiance.  I know swims motivation for replying in such a way is to save both swiy and any other inexperianced bees time money energy and frustration. 

Sterility is the number one stumbling block for beginners IMO, and trying to shoot for shortcuts right off the bat WILL lead to frustration.  It would bee a shame if some newbees read this thread without seeing the conflicting responces your proposal deserves. 

"All that we are is the result of all that we have thought."
-Buddha

bujinkan

  • Guest
Open your mind, and give it a try, you might find ...
« Reply #27 on: September 04, 2002, 09:07:00 AM »
Open your mind, and give it a try, you might find that you can formulate an even better plan
Hear here! good advice. I think, yachaj, that you are being resistant to bulk techniques prematurely....of course to each his own, but birdseed, dung and the like are really not that complicated. THEY DO, require however, experience in being sterile and how to know what is contaminated and what is not. i would go the route all us newb growers go..PFtek, cakes in fishtank terrarium or something, to simple casings or outdoor spawn, to more complicated agar involved strain isolation casing techniques. birdseed and grains will give you many more flushes due to higher nutrient density..period.
the only excuses for using invitro are: experimentation, lack of space, or security. it is effective, but only to a certain degree.
as far as the mushroom spores germinating on caps...well lets just say you should solidify your readings on the matter. i can see where you are assuming things you shouldnt about what is said in studies.  

If I'm reading this correctly, and I like to think that I am.....

goiterjoe

  • Guest
birdseed?
« Reply #28 on: September 04, 2002, 01:05:00 PM »
what's so great about birdseed?  I've seen better results from brown rice flour than from birdseed, although they weren't grown under identical conditions.  Is everyone here recommending birdseed over other grains?

All paths are the same: they lead nowhere

bujinkan

  • Guest
finch seed substrate
« Reply #29 on: September 04, 2002, 01:31:00 PM »
actually mixtures work best: for example finch seed, brown rice and flax seed meal.

http://www.shroomery.org/findorgrowthem.php?View=docs&doc=33


This one aims at complete nutritional balancing using quinoa, flax, finch seed, brown rice and rye.
 

http://www.shroomery.org/findorgrowthem.php?View=docs&doc=117



Birdseed is most useful for spawn, but makes a great substrate if you have a pressure cooker.

http://www.shroomery.org/findorgrowthem.php?View=docs&doc=117



If I'm reading this correctly, and I like to think that I am.....

GOD

  • Guest
swim is, most definatly.
« Reply #30 on: September 04, 2002, 01:34:00 PM »
swim is, most definatly.
Thing is with birdseed, is that the seeds have that thick, hard shell- ya gotta get the shell soaked, so that moisture can peirce through it, and thus make it so that the insides are exposed to the moisture so that when its pressurecooked/sterilized, the insides get hit properly.  Too much moisture, and the seed becomes mush making it clumpy and hard to airate, also making it more prone to bacterial contams.  Too little moisture, the inside stays dry and doesnt get sterilized.  Throughout the whole process, its vitally important to make sure theres an even distribution of moisture for said reasons, that means bee anal mixing.  Swim stopped his mycology experiments a while back, and no longer has his notes regarding amounts used- but from what he remembers, he used pennin#$@n brand finch seed from wallys whacky fun place, he didnt feel the need to remove the sunflowerseeds, although they could bee skimmed off the top during the presoak (remember to stir).  He initially got his seed, did a presoak, keeping track of how much water was used for the presoak (simmering ~45min to an hour keeping the seeds covered in simmering water) swiy will probably have to add h2o as it soaks- the seeds will expand quite a bit- strained the seed, and rinsed it with cold water (or else itd bee too sticky with sugar all over it and itd bee more likely to clump up- also added a small amount of crushed gypsum, about two table spoons to 9L of substrate as a pH buffer and to help clumpyness) then he p.cooked ~1 hour 15 min to 1 hour 30 min.  He then took 100 or 1000 grams of this, and baked it in the oven until absolute dryness.  He subtracted the weight and got the moisture content.  He used the moisture content required that was listed in stamets book- and then added coarse vermic to adjust and achieve proper moisture levels.  Then, when he went back to start making up the substrate for real, hed add a little less water to the pot for the presoak, and add that quantity to the vermic so that it wouldnt bee dry when it went through the p.cooker- just a small amount is needed for this.  Bee careful when pre-soaking, if swiy sees exploded kernals, its a good indication that he isnt stirring enough, remove them if swiy sees them as these make for clumpyness and bacterial contams.  Remember not to pack the substrate.

and thats all I have to say about that...

"All that we are is the result of all that we have thought."
-Buddha

bujinkan

  • Guest
grain spawn 101
« Reply #31 on: September 04, 2002, 01:42:00 PM »

http://www.shroomery.org/findorgrowthem.php?View=docs&doc=123


grain spawn 101.


If I'm reading this correctly, and I like to think that I am.....

Chicken

  • Guest
Substrate
« Reply #32 on: September 04, 2002, 03:20:00 PM »
A rye grain mixed with verticulite (sp?), has yeilded the best results, VS a birdseed, rice, or soy based substrate.  Liquid innoculation was utilized as well as an autoclave, open flame sterilization of tools, all growth was contained within an inncubator made of a converted chest freezer, with HEPA filtration.  All items were easily obtained at industrial surplus stores.  Rye grain is much superior.  Add some erythromyacin (sp?) (available on internet)to the substrate and you will have better luck yet. All substrate was also autoclaved.   Rye flour doesn't work as well as using whole rye grains.  Try it and you will see.  Always was hands 10 minutes with antibacterial soap, and brush nails.  Always use HEPA filter, quatranary ammonia, and UV light when innoculating.  Always use liquid innoculation.  Growth on AGAR plates is possible and viable way to do a non-clone liquid innoculation.  Grown on A Typic soy broth /w erythoromyacin, and then transfered to liquid inocuilation technique.  You will be suprise with results.  Rye grain leaves al others in dust.


GOD

  • Guest
hows the tan?.... UV sterilization BEFORE ...
« Reply #33 on: September 04, 2002, 03:54:00 PM »
hows the tan?....

UV sterilization BEFORE innoculation, not during.
Im gonna bow outta this thread now, Ive already given my 2 cents and dont feel the need to continually repeat myself.  Hopefully, Im not coming across as my way is the right way- but please folks... its all good and well when you put alot of research into a project like this, but please dont claim to know what your talking about- or offer techniques that you havent first tried yourself.
  Its like the blind leading the blind. 8)  ::)  :(  >:(

ps- regarding storage, get yo ass a container, and dump in the shrooms, then perform a co2 releasing reaction over the container (co2 sinks) (forget how to do it...baking soda and vinegar?) -obviously, shrooms should bee bone dry first.  Test to make certain the container is filled w/co2 by lighting a match and lowering it into the box- if it is full, the match will extinguish due to lack of oxygen.  Seal it air-tight and put 'em in a cool environment- whala, they will store INDEFINATLY.

"All that we are is the result of all that we have thought."
-Buddha
edit last point before I leave:

But the agar can be used to clean (molecular filter) the mycelium after the transfer! The technique is called bottom inoculation, it is very useful in cloning and it goes like this. Cut a small square of agar out of the uninoculated, fresh dish/ jar and place it next to the hole it leaves behind. Now place the piece of transferred mycelium (or a primordium) in the hole, and the square of virgin agar right on top of it. Make sure that the mycelium/primordium still has an entry to some air.

This technique may work for bacteria, but only if said mycelium has already been exposed to the peroxide and developed the proper enzymes.  A sterile environment is nessisary to transfer the culture to the awaiting peroxide treated agar.  At the point of transfer, if a foreighn fungus decides to attach itself to the culture, it can piggyback and grow through right along with the cubensis mycelium.  Your proposed method would work IF the peroxide treated agar specifically killed off all other forms of mycelium save cubensis, but it is not, it only kills off the spores etc...

goiterjoe

  • Guest
ok, next question
« Reply #34 on: September 04, 2002, 09:19:00 PM »
Is there anyone here supporting setting up an operation for cultivating mycelium on agar for extraction?  I attempted this once, but had to disband due to a lack of sterile space to operate with.  It seems like you could run a large setup using this method very efficiently, and the only problem that might occur is with keeping sterility.  This eliminates the need for a fruiting chamber, and the extraction would be very efficient compared to fruiting.

Has anyone tried bulk agar cultivation and extraction?  If so, did you use homemade potato dextrose or was it storebought?

All paths are the same: they lead nowhere

bujinkan

  • Guest
i doubt you could get enough for an extraction..
« Reply #35 on: September 04, 2002, 09:51:00 PM »
i doubt you could get enough for an extraction..its always been that fruiting them is the only way to get an acceptable amount for extraction...especially since psilocin/psilocybin contents are low even in late stage mycelium.

If I'm reading this correctly, and I like to think that I am.....

Vibrating_Lights

  • Guest
links
« Reply #36 on: September 04, 2002, 10:19:00 PM »
Can any one give a link that describes the psilobin production in Mushrooms.  How it happens What are the building blocks the fungus uses.  And especially any gene research reguarding using a biosynth pathway making use of a living orginism and altering it's genes.
VL_

So much game I could sell a hooker some pussy
Vl_

bujinkan

  • Guest
Post 184209 offhand i dont know where to find ...
« Reply #37 on: September 05, 2002, 12:00:00 AM »

Post 184209

(PolytheneSam: "Enhanced Psilocybin Production", Tryptamine Chemistry)

offhand i dont know where to find what youre looking for, but you might find it in this thread, or at least an alternative. :)

If I'm reading this correctly, and I like to think that I am.....

Yachaj

  • Guest
GOD on birdseed
« Reply #38 on: September 05, 2002, 04:32:00 AM »
You have obviously not worked with birdseed.  Although you have done alot of research, nothing beats actual experiance.  Open your mind, and give it a try, you might find that you can formulate an even better plan.

GOD, from someone who is forwarding 2nd hand knowledge about mushroom cultivation and does not really have read the manuals he says he quotes I would expect a less arrogant tone.

I have used birdseed&millet many times as spawn. But as cubensis fruiting substrate it is not superb.

I mean - mix 2 volume parts of vermiculite, 1 volume part of powdered brown rice, 1 volume part of tapwater until it is moist and airy. Put everyting in a 250ml shotglass. Top it off with a layer of dry vermiculite of two fingers deep&cover with aluminium foil. Boil for 1 hour in milkpan, cool down & inject sporewater. After a couple of weeks 15-20 percent of the dry weight of the rice is converted in dried cubensis with unopened caps. Potency = 0.5g of dried mushrooms yield a +2 Shulgin Scale.

Besides the inoculation and harvesting it is all completely maintenance free. Drying is not even needed since the mushrooms can be preserved for 3 months at 4 centigrade in unharvested, not-illegal fresh condition (put the glass in th fridge until you are ready for them). After the glass is put at roomtemp, harvest the mushrooms within 48 hours and leave the glass alone. New shrooms come up soon.

I really do no think that a millet (main ingredient of birdseed) technique can improve all that in simplicity and performance (as is evidenced by your own write-up).

This is especially the case when cubensis genotypes are used which are selected for 'PF Substrate' and grown on it, from spores (not tissueculture), for generation after generation after generation for over a decade.

In short - GOD you have no idea what you are talking about. Keep your subjects to the things you are good at.

Yachaj

bibliopharmacophile

bujinkan

  • Guest
have used birdseed&millet many times as spawn.
« Reply #39 on: September 05, 2002, 04:47:00 AM »
have used birdseed&millet many times as spawn. But as cubensis fruiting substrate it is not superb.
How did you use it as spawn? did you sterilize with a pressure cooker?

If I'm reading this correctly, and I like to think that I am.....