By Admin (Admin) on Friday, June 08, 2001 - 05:17 pm: Edit

posted 03-17-2000 08:43 AM
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Simple Cubensis Growing Technique
Version
3-17-00

Table of Contents
Overview
Necessary Equipment
Procedure
Substrate Preparation
Inoculation
Mycelial Growth
Fruiting and Harvesting
Preservation Methods
Credits and Notes

Overview
A quick description of the procedure
A substrate consisting of brown rice flour, vermiculite, and water is sealed in ½ pint jars and sterilized in a pressure cooker. Spores are added to the substrate using a syringe full of spore solution. The spores germinate, creating a living mycelium that colonizes the entire jar full of substrate. The resulting "cakes" are removed from the jars when fully colonized, and placed in a terrarium until mushrooms begin to grow from the cakes.

Necessary Equipment
Equipment for Substrate Preparation
Hammer and Nail - For poking holes in the metal jar lids. The nail should be about 2 mm (3/32") thick.
Mixing Bowl - Fairly large, plastic or metal mixing bowl for mixing up your substrate in.
Mixing Spoon - A long, sturdy kitchen spoon for mixing up the substrate. Use a plastic or metal spoon, not wood.
Measuring Cups - In 1 Cup, ½ Cup, 1/3 Cup, and ¼ Cup Sizes, for measuring out substrate ingredients.
Vermiculite - A soft, spongy volcanic gravel used in potting soil. A good-sized (1-3 cubic feet) bag can be purchased for about $3.00 at most hardware/garden stores.
Vermiculite
Brown Rice Flour - A flour made from long grain brown rice. An ample bag (1-2 lbs.) can be purchased from some health food stores for $3-4.00.
Bottled Spring water - Can be purchased at most grocery stores.
½ Pint Tapered Wide Mouth Canning Jars - They must be half pint size and they must be tapered (sides of the jar slant outward so that the top is widest part of the jar) so that the substrate cakes can easily drop out of the jar. These can be found in grocery stores, hardware stores, and drug stores for much of the summer and during the fall, and cost about $7.00 for a box of 12 jars. During other parts of the year they may be very difficult to find, especially in this odd size. Good places to check during the off season are hardware stores, especially in small towns.

Aluminum Foil - Enough to make a 6" by 6" (15cm by 15cm) square for each jar.
Pressure Cooker or Pressure Canner - The larger the better. Make absolutely sure that you know how to use it, and that all seals, valves, and safety plugs are in working order. For most people this is the most difficult item to obtain. New pressure cookers start at about $60.00 each, and new pressure canners are start at about $250-300.00 each. Pressure canners are much easier to use and hold many more jars, but can be difficult to find. Both can be found at garage sales, thrift stores, etc. Most people who are into canning will have one. This item is essential for preventing bacteria from contaminating your cakes.
Pressure CookerPressure Canner
Sterile Syringe of Psilocybe Cubensis Spores - Available from various sources for about $5-20.00 each. See http://www.drooldonkey.org for a list of spore suppliers. .
Equipment for Fruiting Chamber
Fruiting Chamber - Aquariums, camping coolers, and large plastic Rubbermaid storage containers are examples of suitable fruiting chambers. See Fruiting and Harvesting for a more complete description.
Perlite - Another volcanic gravel used in potting soil, perlite is white and porous. Also can be purchased for about $3.00 at most hardware/garden stores.

Colander - For draining water out of the perlite. Suitable substitutes include a piece of screen or cloth, or anything that will hold perlite but let water drain out.
Dust mask (optional) - Perlite is puffed volcanic glass. When you work with perlite, it tends to be dusty, creating a cloud of glass dust that can be harmful to the lungs, especially if you are asthmatic. Disposable dust masks can be found in hardware stores, often in the Paint section. They are very inexpensive. They can also be used to prevent breathing our germ filled air all over our your cakes and terrariums when working with them.

Equipment for Glove Box (Optional)
Large cardboard box - Preferably unused and clean, at least 12" tall and at least 16" wide by 16" deep.
Saran Wrap (Kitchen cling-wrap) or Transparent Plastic Tarp - Large enough to cover the top of the box. Cling wrap can be placed in overlapping rows if it is not as wide as the top of the box.
Tape - To hold the box together and to attach the plastic onto the box.
Pair of Dishwashing Gloves - Preferably new and unused


Procedure
Substrate Preparation
Poke Holes in the Canning Lids
Take the lids from the ½ pint jars and use the hammer and nail to poke holes in them. One hole per lid is sufficient, however some people prefer to use 2 or even 4 holes, so they can inoculate the cake in more than one spot. This has the advantage of faster colonization of the cake, but is unnecessary. The disadvantage is that every time the syringe needle goes into the cake, there is a chance for contamination. I recommend putting only two holes in your lid. Make sure that the holes are not too close to the edge of the lid. About ¾" (2 cm) in from the edge is sufficient.

Mix Up the Substrate
Using the appropriate quantities as outlined in the table below, mix the vermiculite and brown rice flour together in the mixing bowl, using the large spoon. Distilled water is best for making the cakes, as the chlorine in most tap water can kill or inhibit the growth of your mycelium. However, I have used tap water before with no harm done to the cakes. Add the water and mix thoroughly. This will take some patience and will probably wear out your arm a bit.

PF Max Fruiting Formula
1/2 c. verm.
1/4 c. brown rice flour
1/4 c. spring water

Add Substrate to the Jars
Using the mixing spoon, fill each jar to about 1/2" (2 cm) of the top of the jar. It is very important that the substrate mix be open and airy, not packed into the jar. Dump it in, shake any excess back into the bowl, but don't pack it down at all. Mycelium will grow best in all those little open spaces.

Seal the Jars
Fill the jar the rest of the way to the top with clean, dry vermiculite.
Tighen lid and cover with foil.
Another way to seal the jars is a method that has come to be known as the Ramsey Seal. For each jar, cut a 6" by 6" (15cm by 15cm) square of aluminum foil. Place the square on top of the jar of substrate mix, and carefully press the middle of the foil square down into the jar until it is touching the top surface of the substrate mix. Don't press any farther or you'll compact the substrate mix. The foil will form a little "pond" in the jar, with the corners and edges of the square hanging out of the top of the jar. Fold and press these down so they are flat against the threads on the outside of the jar. Now, fill the "pond" of each jar with clean dry vermiculite, right up to the top of the jar. Use something flat, like a butter knife or a ruler to scrape off the excess. Put the jar lid and ring on, and carefully tighten the ring, making sure that you don't tear the aluminum foil pressed against the threads of the jar. Your jar is sealed and ready for sterilization.

Sterilizing the Jars
Place the finished jars in your pressure cooker or pressure canner, and cook them at 15psi for 45-60 minutes, according to the instructions for your pressure cooker/canner. Let the entire apparatus cool completely. Do not try to open the pressure cooker before it is cool to the touch, and do not try to speed the cooling process, as a quick change in temperature could crack the jars. Be sure to let the jars cool for several hours, because heat is often trapped in the center of the cake, even if the jar feels cool. This heat can kill your spores if you try to inoculate too soon after cooking. It's best if you can let your jars cool overnight. The jars are now sealed containers of sterile substrate. If you have done everything correctly, they can be stored indefinitely until you are ready to use them.
Optional but not recommended:
Since a pressure cooker can often be difficult to obtain, a common substitute used is simply boiling the jars in a pot of steaming water. The pot is filled until the water is about halfway up the side of the jars, and then a lid is put on the pot to hold the steam in. Boil for at least an hour. This method works to some degree, and I have heard of people having a 100% success rate using it. More common, however, is that some jars will get contaminated with bacteria, since many bacteria can withstand normal boiling temperatures. I have also heard of people getting 100% contamination using this method.

Inoculation

Cleanliness Precautions
Inoculating your jars is the main step where contamination is possible, and thus must be done in as clean of an environment as possible. If the room you're working in is clean enough, you can get away with inoculating them in open air. The needle of the syringe, if not absolutely sterile, can carry bacteria and spores from other molds into your cake, contaminating and ruining the cake. Wash your hands and face with antibacterial soap. Wear clean clothes. Anything in the area of the syringe and jars could contaminate your cakes if it is not clean.

Glove Box (Optional)
If you're concerned about sterility, a good way to accomplish this is to make a "glove box," an enclosed, semi-sealed box with holes for gloves to go through and a see-through top. A cheap, halfway decent one can be built for only a couple bucks worth of stuff. All you need is a large cardboard box, some tape and saran wrap to go over the top of the box, and a pair of new, unused dishwashing gloves. Tape saran wrap over the top and cut two holes big enough for your arms in the sides. Disinfect the gloves and the inside of the box with Lysol spray disinfectant. A small gate can be cut into the side of the box for getting the syringe and jars into the box, or they can be put through one of the arm holes (if you choose not to attach the gloves to the holes).

Oven Inoculation
(Optional-Somewhat Risky)

If you have clean kitchen and a clean oven at your disposal, the easiest way to assure cleanliness during inoculation is to do it on an oven rack. Turn your oven on at the lowest possible setting, and let it heat up. Once it's preheated, pull one of the racks out as far as it will go without falling out of the oven. (Use a rack near the bottom) Have your jars and spore syringe all nearby, ready to go. Place 3 or 4 jars at a time on the edge of the oven rack, and begin carefully innoculating them with the syringe. It's a good idea to have a lighter handy as well to sterilize the needle as you go. Flame the needle until it gets very hot, then carefully squirt a little bit of spore solution (if you can spare it) to cool down the needle before sticking it in the cake. Putting a hot needle into the cake will get burnt-on rice flour all over the needle.

Spore Injection
Once you're ready to inoculate, shake up the spore syringe to get as many spores as possible off the sides of the syringe and into the water. Carefully remove the cap over the syringe needle and slide the needle into one of the holes in the jar lid. Shove it all the way in, so that the needle goes thru the cake itself, yet the tip is visible . Gently squeeze out about .5-1.0 cc of spore solution into each jar, splitting up the amount if you inject through more than one hole (highly recommended). Tilt the jar and swish the solution around to cover maximum area.
Most people suggest using an entire cc of solution per jar.
Be careful that nothing but the jar and substrate touch the needle, and re-cap it immediately after using it to avoid contaminating the needle. Also be careful of using too much spore solution. With spore syringes it can be easy to accidentally push the plunger on the syringe too forcefully and dump out way too much solution. Once each jar is inoculated, it is ready for incubation. There is not need to put tape over the holes in the lid, because the dry vermiculite will keep out any contaminants.

Mycelial (Vegetative) Growth
Incubation
Now the jars are incubated at about 75-85 degrees F, in a DARK place, for several weeks. If you have a room that is constantly kept in this general range, this is a good place to incubate your jars. If not, you will need to find some other source of heat to keep them in that temperature range. Be careful not to use any heat source that could cause fires; a heating pad will usually work, some people have used fish tank heaters submerged in a warm water bath. A good investment here is a thermometer that keeps track of highest and lowest temperatures, so you can see how hot or cold your cakes are getting. If they get too cold, their growth will slow considerably, and if they get too hot, they will lose water and eventually die. (They will usually die if they ever get above 95 degrees F)

Mycelial Growth
The first signs of mycelial growth should appear within 5-7 days. If none appear within two weeks, something went wrong. (Perhaps the cake was not cooled completely before inoculation, and the heat killed the spores, or the spores simply did not make it into the cake.) This type of mushroom mycelium will always be a brilliant white fuzz, often growing in ropy strands. This ropy type of growth is called rhizomorphic growth, and is a sign that the mycelium will probably fruit very well. Any other color of mold, including some less brilliantly white molds (cobweb mold, for example, is white but not so thick, and it does look a lot like cobwebs.), is a sign of contamination. A contaminated cake will not recover and, except in very rare instances, will never produce mushrooms.
A colonizing cake displaying rhizomorphic mycelial growthA completely colonized cake in a ½ pint jarA cake, contaminated with a green mold, in a ½ pint jar
Fruiting (Producing Mushrooms) and
Harvesting
The Fruiting Chamber (Terrarium)
Many different things can be used for a fruiting chamber, including camping coolers, aquariums, and large plastic containers (Rubbermaid brand or similar containers work great). The fruiting chamber must be at least 6-8" (15-20cm) tall, and have enough floor space for the cakes to be arranged with at least 1" (2.5cm) of free space on all sides. Spread the cakes out as much as possible so that the mushrooms have room to grow. If the chamber is much too tall or too large, it may be difficult to keep the humidity high enough. The bottom of the chamber must be able to contain water, and the lid must be somewhat airtight in order to keep the humidity inside high. Light must be able to shine into the terrarium. If you are using a cooler or non-transparent plastic container, you will need to cut a window into the top of it and seal it with some sort of transparent material so that light can get in but humidity can't get out. For this reason, glass aquariums make very nice fruiting chambers if they are kept at the right temperature range.

Birthing the Cakes
Once a cake is completely covered in white mycelium, wait at least 1-2 more weeks before taking the cake out of the jar. When you are ready, and in a fairly clean room, begin transferring the cakes from their jars into their fruiting chamber (described in the next step). Remove the lid of each jar, and dump out the loose dry vermiculite on top. Then, put the lid back over the top of the jar. Slowly turn the jar upside down, so that the cake is resting on the jar lid. You may need to gently tap the jar to knock the cake loose. Take the jar off the top of the cake and then carefully pick up the cake and place the cake on the jar lid, put them both into the fruiting chamber. Once all the cakes have been transferred, you're ready to induce fruiting.

Inducing Fruiting (Producing Mushrooms)

In order to initiate fruiting, three main conditions must be met for the cakes.
First, they need light. Only a dim light is needed. A fluorescent lamp or indirect sunlight is plenty of light. Mushrooms do not gain energy from the light like plants do, but in this particular species of mushroom light sends a signal to the mycelium that it is time to produce mushrooms. A source with a wide spectrum of light, especially containing lots of bluish light (daylight and fluorescent plant lights are very good examples of light with lots of blue) is best, but a low wattage (15 watts is plenty) incandescent light bulb will supply enough light.
Second, they need a fairly high humidity. 90-95% humidity is a good range for fruiting. The best and easiest way to do this is by lining the bottom of the fruiting chamber with damp perlite. A common mistake is to get the perlite too wet, and end up with a swamp of water and perlite that is very difficult to clean up, and will drown the cakes. Get enough perlite to make a 1" (2.5 cm) thick layer on the bottom of the fruiting chamber, and put it into a colander, strainer, or cloth enclosure that it can't slip out of. Wet it thoroughly with normal tap water, and let the water drain out. Then move the perlite into the fruiting chamber and smooth out the surface. You now have a layer of damp perlite that the cakes/lids can be set directly on, and which will keep the humidity in the chamber high enough for the cakes to fruit. By the time your cakes have stopped producing mushrooms, the perlite might start getting a little bit skunky smelling. If you want to reuse it, put it in a baking pan and cook it at 350 degrees in your oven until it is dry. Let it cool, and it's ready to be used again. You can also add some Hydrogen Peroxide to the wet perlite to help it stay clean a bit longer.
Lastly, it is a good idea to lower the temperature range a bit, to about 70-80 degrees F. Like the light, this signals the cakes to begin fruiting. However, most strains of Psilocybe Cubensis fruit so easily that lowering the temperature is not really necessary.

Pinning, Fruiting, and Harvesting
For the first week or two, the cakes will generally not do anything. Then, very small bumps, called "pins," "pinheads," or "primordia" will begin to grow out of the surface of the cake. These are the beginnings of mushrooms. Many will never grow any larger. However, some will grow until they are full-grown mushrooms. A mushroom is ready to be picked when the edge of the cap tears away from the "stem" (the stem of a mushroom is called the stipe). Often, there will be a thin veil between the cap and stipe. If this is present, you can wait until the veil tears before picking the mushroom. To pick a mushroom, grasp it near the base where it is joined to the cake, and gently twist it until it comes off. Immediately begin the process of preserving it, either by refrigerating it or by drying it, mushrooms will begin to rot immediately. Each cake will produce about 1-3 waves or "flushes" of mushrooms, normally with 2-5 days of dormancy between flushes. After about a month or so of fruiting, most cakes will be spent, and will not produce any more mushrooms.


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"I dream the same thing every night. I see our freedom in my sight."

[This message has been edited by Hippie3 (edited 03-17-2000).]

[This message has been edited by Hippie3 (edited 03-17-2000).]