PF Jar inoculation and prep

INOCULATION OF THE PF SUBSTRATE JARS

Any jar to be inoculated must be cool to the touch before proceeding. Make sure the lid is tight. Shake the syringe well and remove the tape from the syringe needle guard. This shaking of the syringe is important as to redistribute the spores in the water. Take off the tape covering the needle holes. Remove the needle guard and insert the needle through the lid hole. Tilt the syringe body back towards the center of the lid with the needle tip touching the glass. This distributes the spore water down the side of the jar, giving a good inoculation down the side of the substrate cake. Inoculate a few drops down each needle hole. As the syringe plunger is pressed, observe the needle tip against the inside of the glass. As soon as water appears around the needle tip, release the syringe plunger pressure. In between each hole inoculation, shake the syringe a little to keep the spores distributed. Use 1 cc per jar. This will allow the syringe to inoculate 10 jars. More spore solution per jar can be used (speeds colonization), but fewer jars can be inoculated . If the syringe needle plugs up as it is inserted into the substrate, draw the needle back a little and it will unplug.

In this photo, the needle tip can be seen resting against the inside surface of the jar. Then, when the solution is injected, it will run down the side of glass, giving an even inoculation. It is also important to add, that the vermiculite in this jar photo is very course. This makes the needle more visible for the demo. This type of vermiculite is best avoided.

ALCOHOL FLAMING TECHNIQUE

If the syringe needle is touched, flame the needle to sterilize it. An alcohol flame is a clean flame whereas a butain cigarette lighter leaves behind an undesirable soot residue. To produce a short burning alcohol flame, place a tequila shotglass upside down. Using an eyedropper, put a few drops of denatured alcohol fuel (hardware store) on the hollow bottom of the glass and touch it with a match or lighter. The blue flame will cleanly and safely sterilize small stainless steel tools. Heat the needle in the flame for a few seconds to resterilize it. There might be a few "pops" of boiling water spurt out of the needle, but the spores within the syringe are safe. If there is some left over spore solution, replace the needle guard and store the syringe for later use. Resterilize the needle immediately before re-use. Store the syringe in a dark, cool place.

INOCULATION OF PF JARS WITHOUT THE LIDS

This technique can also be used if canning jars are not available (1/2 pint wide mouth canning jars are perfect and should be used at all cost). If regular drinking glasses are to be used - use regular tapered sided drinking glasses (8 ounce - 250ml)

Jars can be inoculated without using a lid with holes punched. Before trying this technique, inoculate with the punched lid first. That will show how it works without any problems (almost fail proof).

The only precaution to observe is to disturb the dry top vermiculite layer as little as possible, especially when removing the needle after the inoculation. The underlying substrate must not be exposed to the air. Carefully move any disturbed vermiculite back into place. If using a drinking glass or alternate container, cover the mouth with tin foil. Replace the tin foil cover after inoculation.

INCUBATION OF INOCULATED JARS

After inoculation of the jars, tighten the lid bands and retape the needle holes. Place the jars in a safe place out of direct sunlight. Indirect light is all that is required. If the temperature is kept around 70 degrees, germination will begin within 3 to 5 days. Germinating spores appear as small white fuzzy spots, quickly growing and spreading with cottony white growth and strandy "rhizomorphs". Any room temperature is O.K. If it gets cold indoors, over head light shinning down on the tops of the jars is a perfect heating technique for this culturing stage. A clamping type light with a reflector works well for this. If this is done, keep the temperature around 70 degrees (don't overheat the jars - monitor the temperature with a thermometer). A warm overall house temperature is fine. But in the overall view, cool temperatures are never a problem. The rule is to not overheat.

THE CANNING JAR LID (loose or tight)

There are two choices with the lids during incubation - tight or loose. With a very high moisture content (good for fruiting), a tight lid can cause water to collect in the bottom of the jar. This is to be avoided. If it happens, the lid should be kept on loose during incubation. Tape the canning jar lid to the band to make the lid act as a one piece lid for raising and lowering. If the substrate is on the dry side, a tight lid will preserve the moisture content. It is all a matter of the balance between the water needs of the mycelium, the size of the jar, the available air space in the jar and the type of vermiculite used. Only by simple experimenting and comparison can the right balance be found for a given set of conditions. Take notes and go with what fruits the best.

After the substrate turns white with the mycelium (2 or 3 weeks after inoculation), the jars are left to sit in indirect light. The mycelium will continue to infiltrate the substrate until it gets enough food to trigger the fruiting cycle. In less than a week to a few weeks after surface colonization of the cake, tiny white "pin" like structures begin to appear. This is called pinning. This is the beginning of the fruiting cycle. Soon after that, within the week, small round fungus growths appear that soon begin to turn yellow.

Lastly, "primordia" start to grow. These are tiny worm like structures with tiny reddish heads. These are the first mushrooms.