The methods described in this manual require very little in the way of equipment for growing your own mushrooms at home. You will NOT need a glove box, HEPA filters, ultraviolet lights, a sterile laboratory, laminar flow hoods, air locks, foot washes, etc. etc.
As in the first volume, most of the methods in the second volume have been designed primarily for small scale and home cultivation. But I designed the first two methods for preparing bulk substrate specifically for growers who want to work at commercial scales. And nearly all of the remaining techniques presented here could also prove useful in a commercial context.
The peroxide-based methods presented in this volume are all my original inventions. In general, the non-peroxide methods have been worked out by others and have made there way into the public domain. I am presenting them here because of their obvious value to mushroom growers who use the peroxide method.
I've noticed a few people lately asking about agar. I find it very useful and recommend that everyone give it a try. Most people are turned off of agar because of fears of unavoidable contamination problems. I've found that with the use of hydrogen peroxide in my agar mix, I can do transfers in the open, unfiltered air with little worry of contamination.
Many people seem a bit confused as to what peroxide can and can not do. Hydrogen peroxide will kill bacteria and fungal spores and inhibit the growth of yeasts. Unfortunately, mushroom spores fall under the category of fungal spores, so they will not germinate on peroxidated agar because they are killed on contact. The good part about this is that mold spores, such as spores from the dreaded green mold, fall under this category as well and are also killed upon contact with the agar surface. What will grow on peroxidated agar is existing fungal colonies, such as your living mushroom mycelium, because their cells can produce enzymes that will cause the peroxide to be broken down into water and oxygen. The downside, however, is that existing colonies of some molds, such as green mold, are also capable of producing these enzymes, so peroxide will not guard against mold spores that have already germinated. But perhaps the best attribute of peroxide is that it will kill all bacteria, so you will never have a problem with bacterial contamination on peroxide enriched agar.
... mix three grams of malt extract and three grams of agar with 149 ml of water (you should use 149 ml instead of 150 because you want to account for an extra ml of peroxide that you will be adding later)...
A range of H2O2 concentrations is acceptable. Cubensis seems to tolerate amounts ranging from 1 ml to 7ml for this mix. The more you use, the longer "adjustment" period for mycelium to adapt to the peroxidated agar. As a general rule, the least amount necessary is best. But since your mileage may vary, you may modify as to what works for you.
I have personally used the original formula to make plates that were left unsealed and unfiltered in a nasty NASTY kitchen for up to 2 weeks until they dried out. The agar never contaminated!
Bulk Neglect Tek
Cloning: As such, a chunk is cut from inside [the mushroom's] stem near the base. The chunk is rinsed in 3% H2O2, and placed in a small jar containing sterile distilled water and peroxide (roughly .0015% H2O2). A filter disc in the lid of the jar allows the gasses produced by the peroxide to be vented. The jar is left at room temperature until the initial harvest, drying and bio assaying are completed. A period of roughly 2 weeks passes. Peroxidated MEA is prepared in the standard manner. Peroxide concentration is on the higher end of the recommended levels. The chunk is removed from the jar and placed on the agar. The plate is mostly colonized in a week and a half with temps hovering near 70 deg F.
Spawn: A quart of birdseed spawn is selected from several prepared in the following manner. Birdseed rinsed clean several times in cold water (which btw removes a lot of sunflower seeds if you arenít careful, or want it to, sunflower seeds float and are easily poured off from the rest of the BS). Boil an excess of water first. Reduce heat once boiling. Seed is simmered on low for 30 minutes (no more since we will be adding peroxide later and donít want too much moisture). The grain is strained, rinsed and dried thoroughly before being loaded into the quart jar, half full. The lid has a filter disc in it, but polyfill, or even coffee filters or cardboard can be substituted since we will be adding peroxide later. The jar is pressure cooked to sterilize and also destroy peroxide decomposing enzymes. When cool, the quart is injected with 12 ccís peroxide, shaken and inoculated in front of a flow hood with a piece of the peroxidated agar. Colonization is complete in a week with temps around 80 deg F. A huge filter patch bag is prepared with birdseed as above. Peroxide amount is proportional to that listed above. In the flow hood, the bag is inoculated with the colonized quart jar and sealed with an impulse sealer. The bag is colonized in 11 days at room temps.
Sterile Tissue Cloning Procedure
1) Create a paste of brown rice flour & water (or your choice of substrate), spread evenly in the bottom of a canning jar, seal, pressure cook at 15 psi for 30 minutes.
2) Pour about 1/3 cup of 3% h2o2 into a jar.
3) Obtain a clean coffee grinder; fill with water to the brim of the metal cup, insert a fresh and cleaned mushroom stem (no cap) and proceed to grind for 10-15 seconds.
4) Use a clean spoon to transfer some of the resulting mixture into the h2o2 jar.
5) Let it fizzle for a couple minutes, then draw into a syringe.
6) Inoculate jar created in step #1, or simply inoculate pf-style jars.
Peroxide Mushroom Cultivation Method (edible mushrooms)
Fill a cooking pot with 3-1/2 cups of water. Cover, bring to a rolling boil. Reduce heat, remove the pot after 1 minute.
When the pot of boiled water has cooled to room temperature, lift the cover and stir in 2 tablespoons of 3% hydrogen peroxide solution, available at all drug stores and markets (the brand name is not important). Now, pour the water/hydrogen peroxide mixture into the bag. Again, close the bag loosely and allow it to cool to room temperature.
Beating Green Mold (shroomery forum)
these casings had the green really bad when i birthed it =)
i beat the green-
all these casings had it bad when birthed-sorry no flash-batteries was low-kinda dark-
nothing but h202,and water was used- =)
Peroxide Mushroom Growing FAQs
Peroxide can do away with costly filter-patch culture bags for bulk substrate. Grow cultures in ordinary trashbags (placed inside boxes) right out of the package, or in reusable plastic buckets with lids
Peroxide kills mushroom spores, so you can grow agar cultures in the same building you use to fruit your mushrooms, even if the mushrooms produce a high spore load.
A small quote from Rush Wayne's Peroxi Manual Volume II, It involves using Grey Cardboard instead of agar -- detailed steps for making cardboard plates. Note that you can also use small jars in place of Petri dishes.
Peroxide and Straw
Despite this complicated explanation, the protocol for preparing straw with peroxide is extremely simple. It goes like this:
1) Place your straw in a large soak vessel.
2) Fill the vessel with the appropriate cold solution (see article) to immerse the straw.
3) For chopped straw, soak about 4 hours at room temperature. For unchopped straw, soak for at least 28 hrs, or until the leachate takes on the color of a good tea.
4) Drain the straw thoroughly, until it is no longer drippy.
5) Remove the straw to your culture containers, mixing in spawn as you go.
I inoculate the straw by filling a plastic trash bag, draped inside a large cardboard apple box. I put in a layer of straw, then a sprinkling of spawn, then another layer of straw, then more spawn, and so forth. Finally I close up the bag and compress the straw by pushing the bag and its contents down into the box as far as I can get it to go. A quarter of a bale of straw fills three apple boxes worth. The bags get loosely twisted closed and covered with old bed sheets to incubate for about a month.
Cloning Live Mushrooms to Peroxide Agar
Take the shroom and clean it with the iodine, then slice it open and remove a chunk of tissue from the inside, then quickly tranfer it to the peroxide agar.
Hippy3's Bleach Experiment (shroomery forums)
So far as trich goes, bleach solutions can work, but getting the level where you can knock out the trich without harming the mycelium is tricky. It's a pretty fine line. But then again, damaged mycelium is better than trich infestation. I first used a bleach solution (10%) on trich on accident. It killed the trich, but seriously affected the casing as well. Later experiments with more dilute solutions worked better, but always had an impact on the myc at strengths great enough to battle the trich. This was on casings of course, it may be easier on cakes.
Q: i thought the ratio is 100/1. i haven't dunked in bleach yet but i'm going to try with my current batch. is it 200/1 ir 100/1 for dunking?
A: I have had great results with 100:1 but most of the posts I have seen on the subject are from people who had problems with that concentration. I only use 100:1 after the first flush when I start seeing little contams poking their nasty little heads up. Although I haven't tried it more people have claimed overall success from a 200:1 ratio. Use the 200:1 for now and if it doesn't kill the meanie greenies then bump it up slowly.
A: exactly, for most purposes it makes little difference, but some did report that the higher concentration, esp. at warmer temperatures, damaged and stunned the mycellia so much that contams easily returned even worse. so i recommend the lower concentration instead, it's still more than enough to kill germs. besides, the best way to kill contams is not stronger concentrations, but instead multiple dips in fresh solution.
Bleach Dunk Tek Results (Mycotopia forums)
ok so this guy i know tried the bleach tek on his last bunch of pf cakes....and heres the astounding results: here's PF stropharia's that got the bleach tek for approximately 5 minutes after a 6 hour spring water dunk. the bleach solution was a very dilute mix, 2 capfuls of bleach in a gallon of tap water
Quick Bleach Questions (mycotopia forums)
If you're talking about bleached cakes, I've noticed that my cakes after bleaching take longer to pin, they grow a thick layer of white dense mycelium - then begin to pin.. but I've also noticed some of my un-bleach-dunked cakes have pulled the same stunt - taking several (7+ days) to pin..
You might want to back your solution down to 1:200 as well. And if you see trich, cut it out and get rid of it quick. It will spread to the other cakes it you don't kill it quick.
"Bleach dunk killed my cakes" (mycotopia forum)
I have been using about 8 mml of bleach per gallon and it disinfects perfectly for dunking 10 to 20 minutes after plain water dunk for 24 hours in the fridge. For those who are skittish about using the 100:1 solution this solution is less that 200:1 and works fine.