Author Topic: Mushroom surface sterilization  (Read 12569 times)

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SaintCyril

  • Guest
Mushroom surface sterilization
« on: August 23, 2002, 03:17:00 PM »
In growing mushrooms the most difficult part is reduction of bateria and fungus wich might over colonize the mushroom growth in the early stages.  Obviously washing hands for 10 minutes with antibacterial soap and using an autoclave for the media and all tools, and glassware, ect.  But what I have noticed is a many use bleach or somesort of other chemical to sterilize work surfaces and the inside of the incubator.  I used to use bleach, but switched to quatranary ammonia a few years ago, and the stuff works wonders.  In a bio-med lab we just called it quat, its blue and works very well on surfaces to eliminate unwanted bacteria and fungus.  I suggest it in addition to the above methods to anyone, and to a newbee it will reduce the unwanteds much better on tools if you happen to not have an autoclave.

Cy

We are the people that your parents warned you about.

GOD

  • Guest
if there is no access to an autoclave, a pressure ...
« Reply #1 on: August 23, 2002, 03:56:00 PM »
if there is no access to an autoclave, a pressure cooker and Al foil are your best friends, dont leave anything to chance.

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

SaintCyril

  • Guest
Autoclaves
« Reply #2 on: August 23, 2002, 04:11:00 PM »
If you go to an industrial surplus store in your town they will almost for sure have a medical grade autoclave in stock for very inexpensive, less even then the nicer stereoclaves.  Industrial surplus stores are my best friend, and even though you don't know it if you are in an area with more then 100,000 population there is probably one in your town, and some of them are like junk yards, they are linked up to other industrial surplus stores, and can get whatever you want in a couple of days.  Ussual security precautions, fake name, fake corporation, fake mailing address, fake ID, would probabnly be in order if you are the paranoid type, but seriously I have never had them ask for any of that info unless it was something I was ordering, and I had to put $$ down.

Cy

We are the people that your parents warned you about.

Yachaj

  • Guest
obsolete mushroom cult techniques
« Reply #3 on: August 29, 2002, 04:48:00 AM »
I hope there is no need to post this statement more than once:

YOU DO NOT NEED A PRESSURE CANNER OR STERILE ENVIRONMENT FOR THE CULTIVATION OF FUNGI. Period. Nor do you need to wash your hands or clean surfaces. Do not believe anyone who states otherwise for they are twenty years behind in knowledge or (most likely) they want to sell you expensive equipment (laminar flow hood, pressure canner, surface sterilization liquids, complete sterile rooms etc. etc.)

You only need to do that if you are too tight assed to invest in 3 manuals: the Psylocybe Fanaticus TEchniques (PF TEK) and 'Growing Mushrooms with Hydrogen Peroxide' vol. I and II.

Yachaj

bibliopharmacophile

bujinkan

  • Guest
YOU DO NOT NEED A PRESSURE CANNER OR STERILE ...
« Reply #4 on: August 29, 2002, 12:19:00 PM »
YOU DO NOT NEED A PRESSURE CANNER OR STERILE ENVIRONMENT FOR THE CULTIVATION OF FUNGI. Period. Nor do you need to wash your hands or clean surfaces. Do not believe anyone who states otherwise for they are twenty years behind in knowledge or (most likely) they want to sell you expensive equipment (laminar flow hood, pressure canner, surface sterilization liquids, complete sterile rooms etc. etc.)

Im not sure what youre getting at here. sterility is key with fungus my friend..sure you can do PF tek with stovetop pastuerization, but pressure canners are always better.
as far as your statement about cleanliness and *not washing your hands*, thats pretty silly. the only way this is so is if you are doing outdoor growing, and even then you need to get sterile spawn.

Im not trying to be contrary, but ive seen lots of growing setups, lots of substrates and techniques. by far the highest yeilds come from casings.  usually 5050 (verm/peat) spawned with finch seed. Im not sure what technique youre getting at...im assuming some invitro pinning stuff. Thats fine...but if you  really want to grow shrooms, and i mean lots of them, learn how to use birdseed spawn....and yes for that you do need a pressure canner(finch seed by nature is next to impossible to sterilize without a canner).

ok, so you dont have to take my word for it. heres some examples, courtesy of my friend ryche hawk.
thai cubensis

http://www.thehawkseye.com/thai_lipa/thaily1_1fl.jpg


cambodian cubes

malaysian cubes

http://www.thehawkseye.com/malaysia/mal_bs3.jpg


panaeolus cyanescens and a few 'b' strain PF cubes

golden teachers

http://www.thehawkseye.com/eq/ecu_od2_4.jpg



all these casings are from this technique:

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


except for the pan cyans, which im not too sure about. Ryche does have a grow room with hepa filters, but this isnt totally neccessary. the important thing is the substrate, the sterilization, and the growth environment.


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

bujinkan

  • Guest
yes,
« Reply #5 on: August 29, 2002, 12:24:00 PM »
yes you have to wash your hands. cleanliness is key. Humidity, temperature, air exchange are also key for eliminating and controlling contaminants, as well as optimizing growth.
'Plantasia Mystery' cubensis


BTW: this probably doesnt belong in Tryp Chem.
Thai (lamai strain) cubensis


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

paranoid

  • Guest
Yachaj
« Reply #6 on: August 29, 2002, 03:56:00 PM »
=RATED AS - Idiodic=


"obsolete mushroom cult techniques    

I hope there is no need to post this statement more than once:

YOU DO NOT NEED A PRESSURE CANNER OR STERILE ENVIRONMENT FOR THE CULTIVATION OF FUNGI. Period. Nor do you need to wash your hands or clean surfaces. Do not believe anyone who states otherwise for they are twenty years behind in knowledge or (most likely) they want to sell you expensive equipment (laminar flow hood, pressure canner, surface sterilization liquids, complete sterile rooms etc. etc.)"


Any newbees reading this please disregard, this guy is obviously either a moron or fucking with you.
 

GOD

  • Guest
yach- tell me this then, with the peroxide...
« Reply #7 on: August 29, 2002, 05:10:00 PM »
yach- tell me this then, with the peroxide... How the hell are you going to get the spores to germinate?  Either way, there is no way around sterile procedure.  Swims seen that manual too- read the thing before spreading misinformation!
How the hell are ya going to do the clean, sterile work on them dishes without means to sterilise PROPERLY.  Newbees, shortcuts=contamination.  There is no avoiding having to acquire the proper tools and the use of sterile technique.

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

goiterjoe

  • Guest
bujinkan's photos
« Reply #8 on: August 29, 2002, 10:10:00 PM »
are those amanitas I see growing in the second photo with the attached veils and huge stalks, or are they just extremely large cubensis mushrooms in really bad lighting (I should sober up and look at these tomorrow)?

All paths are the same: they lead nowhere

ClearLight

  • Guest
He's waiting too long...
« Reply #9 on: August 29, 2002, 10:50:00 PM »

  The caps are already open... he'd have stronger shrooms if he harvested earlier...


Infinite Radiant Light - THKRA

GOD

  • Guest
Those are cubensis, not amantias.
« Reply #10 on: August 30, 2002, 09:13:00 AM »
Those are cubensis, not amantias.

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

Topologist

  • Guest
Pine trees
« Reply #11 on: August 30, 2002, 10:24:00 AM »
Unless you have a pine tree in your living room aminitas are 100% impossible to grow in a lab condidtion, or indoors for that mater.  Yest thats right i said 100%, meaning no one has ever done it or ever will, because the aminita mushrooms are symbiotic with pines, and need the pines root system to live, and yeas the tree has to be healthy and alive.  Heard of some guy with a christmas tree farm growing them before. . .


bujinkan

  • Guest
goiterJ, those are "b+" strain cubensis.
« Reply #12 on: August 30, 2002, 01:41:00 PM »
goiterJ, those are "b+" strain cubensis. theres no known  way to get amanitas to grow indoors...and even if there was it probably wouldnt be worth the effort.

clearlight, he's a spore vendor so he is probably going to print all those caps...and if not he's probably just letting them open for the pictures sake.

btw, yachaj..i dont understand how or why you posted this. you seem knowledgable in other things. you recommend PF tek, which is where everyone learns about the absolute necessity for sterility. it doesnt need to be 'ebola lockdown' sterile, but you still have to be very very careful not to introduce microbial competitors to your substrates.

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

foxy2

  • Guest
nope
« Reply #13 on: August 30, 2002, 01:55:00 PM »
buj
Those are his friends pics and pics sent to him.  He doesn't grow the mushies.  Didn't you ever read the bust story he had up a year or two ago?  He got raided and they stole tons of his shit but he only had a tiny amount of mushies, like 5grams or something.  They still took his laminar flow hood tho. :(

Those who give up essential liberties for temporary safety deserve neither liberty nor safety

bujinkan

  • Guest
hawk
« Reply #14 on: August 30, 2002, 08:30:00 PM »
i heard something about that...

My impression was that him and friends had secret growrooms and that they worked together on. who knows. in any case, the technique used here is overwhelmingly the 5050+ casing tek..and id be willing to bet that most if not all of the indoor pics are direct from ryche and friends using his techniques. (this, because of videos of growrooms ive seen on the shroomery...pink floyd in the backround, the whole 9 yards)

i didnt know that they actually raided his place though...i thought that they were trying to shut him down on legalities that didnt exist. In any case someone is producing well for him: i got a few packed syringes a month or two bfore i cut out.

and if youre trying to keep his name out of the light...its a little late for that...and in this corner of the web it matters even less.

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

Yachaj

  • Guest
The 10 bad karma points bet!
« Reply #15 on: August 31, 2002, 07:02:00 AM »
In reply to paranoid and bujinkan:

Bujinkan wrote:
>Im not sure what youre getting at here. sterility is key >with fungus my friend..sure you can do PF tek with >stovetop pastuerization, but pressure canners are always >better.

No. Not if you grow only small batches of mushrooms. True, a pressure canner can sterilize a halfpint jar of pelletized substrate in 20 minutes. A milkpan can do it in 60 (at sealevel). But a pc needs 20 mins to heat up and 20 to cool down to the level that the lid can be opened.

The pics you show are obviously of bulksubstrate grown mushrooms. Lemme tell you some things about bulksubstrate:

- it is sensitive to contaminations
- it is a lot of work
- daily maintainance is required (watering!)
- you end up with far more mushrooms than needed for an experiment (legally risky and it usually means that a harvest needs to be stored for a longer time. Potency declines)
- biological efficiency is *low*. Where 16-20 percent of the dry weight of a mixture of powdered brown rice, vermiculite and water will be converted in dry cubensis mushrooms (with an alkaloid content of of 0.7-1.0 percent), bulk substrates show an efficiency which is half in harvested weight and an alkaloid content of 0.7 at best.

Bulksubstrate cultivation is for shroom dealers (who get their money by selling mushrooms by their weight), not psychonauts (who grow their mushrooms for alkaloid content).

A pc is indeed handy for the sterilization of whole grain spawn, especially in amounds which exceed a half pint. But then again, only bulk growers like RHawk do need that. I do not and neither do you. Growing mushrooms in bulk means a pile of legally risky endproduct, with a potency which is both too low and declining.

To paranoid:

Believe me - you do not need a pc or HEPA blower for the preparation/inoculation/incubation of media. I know because I do it all the time. Also in bulk (not cubensis or another psychoactive species but, at this moment, the delicious Hericium erinaceus.

All the necessary techniques are in the manuals I mentioned but I do not give details here because I know it took the authors of those manuals a loong time to design the techniques. I do not want to hurt the sales they deserve but I hereby state again that for germinatin spores, preparation of media, spawn, bulksubstrate no pc or hepa blower or clean lab is necessary. And you do not ned to wash your hands.

But hey, mr. Paranoid: I do no want to go along with you and make this thread into a flame war, but since you called me a moron I bet for 10 bad karma points that I am right. OK?
 

bibliopharmacophile

Yachaj

  • Guest
Spore germination without pc, peroxide or flowhood
« Reply #16 on: August 31, 2002, 07:28:00 AM »
yach- tell me this then, with the peroxide... How the hell are you going to get the spores to germinate?  Either way, there is no way around sterile procedure.  Swims seen that manual too- read the thing before spreading misinformation!

GOD (or SWIG), I suggest to read the manuals more closely before you accuse me of spreading misinformation. The spore germination technique (no peroxide, no hepa/glovebox) is at page 7 of volume II of the peroxide manual. The technique suggests a pressure cooker but from personal experience I know that it is not needed.

A spore germination 'technique' which is not mentioned in any of the manuals I mentioned is the following.

Grow mushrooms with the PF TEK and do not harvest. When the mushrooms become over-ripe lots of spores will collect on the top surface of the caps - and germinate!

From there, Waynes' peroxide techniques can be used. But the technique for making spore syringes in the PF TEK itself is even easier. Spores are germinated on the fruiting substrate!

A bulk version of the germination-in-endsubstrate technique (for outdoor purposes) is detailed in Growing Gourmet and Medicinal Mushrooms of Paul Stamets.

bibliopharmacophile

GOD

  • Guest
YOU DO NOT NEED A PRESSURE CANNER OR STERILE ...
« Reply #17 on: August 31, 2002, 09:48:00 AM »
YOU DO NOT NEED A PRESSURE CANNER OR STERILE ENVIRONMENT FOR THE CULTIVATION OF FUNGI. Period. Nor do you need to wash your hands or clean surfaces. Do not believe anyone who states otherwise for they are twenty years behind in knowledge or (most likely) they want to sell you expensive equipment (laminar flow hood, pressure canner, surface sterilization liquids, complete sterile rooms etc. etc.)

Yachaj-
  First off, no offense was intended by my post.  The main thing that I find misleading about the above statement is where it says that no sterile environment is need to grow mushrooms.  Having known people who consider themselves BOTH psychonaughts AND have grown bulk for quite a long period of time, I cant help but assume that they know what they are talking about.  What it boils down two are 2 different stages in the growing process, well one actually- and that is where the substrate is innoculated.  If it is through spores, peroxide is gonna kill them off- thats why peroxide is used.  Youd have to bee super lucky to bee able to take a sporeprint, make a syringe etc... and not get anything else in there thats gonna fuck up the process (other fungus/bacteria piggybacking).  Same holds true when it comes time to clone. 
  Those are the only 2 ways- to date that your gonna bee able to innoculate substrate.  Substrate as well- unless your working outside, for indoor cultivation, which the methods you proposed are methods of- substrate needs to bee sterile, cultures need to bee sterile, spore syringe needs to bee sterile until it is holding UNCONTAMINATED spore solution.  The only way to introduce uncontaminated spores to a sterile substrate is through sterilisation of the substrate, and using peroxide to do this (with spores) will not work- the peroxide method can bee used in this manner if one is using cloned mycelium that has been allowed to grow on peroxide treated agar-  BUT in order for one to ensure that nothing is piggybacking the mycelium, the clone needs to bee taken in a sterile environment, and allowed to grow out on the peroxide treated agar in a sterile environment or else it WILL pick up other forms of fungus that float around in the air. 
  A sterile environment is nessisary UNLESS you are going to make 10,000 attempts and you plan on growing out that one attempt that just happens not to pick up a foreighn species.
  The bane of mushroom cultivation is contamination, and the only way to avoid it is through a sterile environment.  It cannot bee avoided.  And no, I am not trying to sell any expensive labware  :P .  Just speaking from second hand experiance  ;) .

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

bujinkan

  • Guest
, yachaj? /
« Reply #18 on: August 31, 2002, 01:18:00 PM »
like RHawk do need that. I do not and neither do you. Growing mushrooms in bulk means a pile of legally risky endproduct, with a potency which is both too low and declining
Potency does not decline as a result of more nutrients. not only does your statement defy common sense, it also defies the experiences of many many growers.
No. Not if you grow only small batches of mushrooms. True, a pressure canner can sterilize a halfpint jar of pelletized substrate in 20 minutes. A milkpan can do it in 60 (at sealevel). But a pc needs 20 mins to heat up and 20 to cool down to the level that the lid can be opened.

A pressure cooker sterilizes more thoroughly. period. No one is trying to be higha nd mighty in telling you this...i myself have used stovetop pastuerization manytimes and it works pretty well, but id still rather have a pressuer cooker.
Lemme tell you some things about bulksubstrate:

- it is sensitive to contaminations
- it is a lot of work
- daily maintainance is required (watering!)
- you end up with far more mushrooms than needed for an experiment (legally risky and it usually means that a harvest needs to be stored for a longer time. Potency declines)
- biological efficiency is *low*. Where 16-20 percent of the dry weight of a mixture of powdered brown rice, vermiculite and water will be converted in dry cubensis mushrooms (with an alkaloid content of of 0.7-1.0 percent), bulk substrates show an efficiency which is half in harvested weight and an alkaloid content of 0.7 at best.

-as compared to what? cakes? this is where the cleanliness factor comes in.
-daily maintanance is required with any shroom setup except sclerotia and invitro. You need air exchange. as far as watering goes, well you need humidity...theres automated setups.
-not everyone wants shrooms for 'experiments'
-THis last one..=)... you need a source to back this up. this is a bold statement...goes against logic and reasoning as well.
 Bulksubstrate cultivation is for shroom dealers (who get their money by selling mushrooms by their weight), not psychonauts (who grow their mushrooms for alkaloid content).
not really. those shrooms i posted are probably better than any shroom either of us have ever had. I fail to see how a shroom can produce maximum alkaloids with very little growth medium.
All the necessary techniques are in the manuals I mentioned but I do not give details here because I know it took the authors of those manuals a loong time to design the techniques. I do not want to hurt the sales they deserve but I hereby state again that for germinatin spores, preparation of media, spawn, bulksubstrate no pc or hepa blower or clean lab is necessary. And you do not ned to wash your hands
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


Grow mushrooms with the PF TEK and do not harvest. When the mushrooms become over-ripe lots of spores will collect on the top surface of the caps - and germinate
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.
I dont know where you get your info from, but you need to check your sources..seriously. :-[

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

Yachaj

  • Guest
Mycrosterile puzzle
« Reply #19 on: September 01, 2002, 03:36:00 AM »
First off, no offense was intended by my post.

No Problem. I didn't take it as such. And GOD I like your contributions to the MHRB purification technology a lot. But non-lab mushroom cultivation 'in the wild' is my favorite way to spend time. If I have any expertise it is on this subject. I want to share what I know with this community if I am allowed to ask dumb questions about biochemistry now and then.

But enough of this. Back to the zroomz.

The main thing that I find misleading about the above statement is where it says that no sterile environment is need to grow mushrooms

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.

The desired mycelium needs to grow in a container where no other life forms are present (petri dish, sauce jar, test tube, etc.). To be able to introduce the mycelium or spores in that container, the inside of it must have some permeatable entry which is big enough to let through both the spores/mycelium and the carrying utensil but preferably not contaminants (if contaminants come in then they need to die before they can multiply). Furthermore, the mycelium needs to have access to fresh oxygen during incubation.

That is the puzzle. And it is solved in the manuals I named.

That is to say... I see no possibility to introduce a wedge of agar in a new saucejar/petri without introducing unicellular contaminants.

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.

The mycelial hyphae are able to penetrate the agar while growing upwards. Bacteria can not. Result: on top of the agar appears clean white mycelium in a week or so. Ready for transfer.

In the few days that it takes for the mycelium to grow through all unicellular contaminants in the jar/petri have hit the peroxidated agar at least once and have died as a result of it. So the inside of the container is cleaned too, in despite of the fact that it has been opened during inoculation!

In the PF TEK, the filtering is done by the dry vermiculite contaminant barrier. Which is permeatable by the syringe needle (or even a pipette or eye dropper if the layer is thick enough) without exposing the moist substrate to unsterile air. The reason that PF's simple approach for making spore syringes works (print mushroom cap in baked or boiled jar, inject sterile water/suck sporewater in syringe/inoculate new substrate cake) is because of the overkill of spores compared to the contaminants.

If a sporeprint is dark, fresh and used for just a handfull of syringes and th inoculant is spread well over the substrate (4 inoculation hols or more) the contaminants just can not take over fast enough.

The idea which is proposed by the author of the peroxide book to germinate spores (no flowhood, no water, no vermiculite AND no peroxide) is alo uniquely original. It is absolutely amazing that so few people have recognized the potential of it.

Yachaj

bibliopharmacophile