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OfflineShamanWarrior
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Inoculation FAIL or Incubation FAIL?
    #19241313 - 12/07/13 02:45 PM (10 years, 1 month ago)

Ordered 4 syringes from a trusted source, first syringe and round of 12 jars went wonderfully. Used alcohol to sterilize syringe between jars, no flame, and great success. Jars were kept in dark at 82-85 degrees during incubation.

I stored the 3 other syringes in the fridge.

A month later, I inoculated another 12 jars with one of the syringes, same process as first round, only this time I used the flame sterilization technique on the syringe between jars. Let it cool for a few seconds and then gave the squirt. I stored these jars the same as first round, 82-85 degrees during incubation. After several weeks, these jars had ZERO sign of growth. No mycelium, no contams.

About 2-3 weeks later, I did a final round with syringe 3 and 4 on 24 jars, each 12 at different times a couple days apart. On these jars I sterilized half with alcohol swab and half using flame technique. I stored most of the jars in 82-85 degrees in dark, and another 6 jars in my closet, where temperatures are much lower, 60-70 degrees. After several weeks out of the 2 syringes and 24 jars, I have only 2 jars actually showing any signs of mycelium growth, growing very, very slowly. 1 of those jars is in the dark 82-85 degree spot, the other in the closet at 60-70 degrees.

At first I was suspicious whether or not the syringes were bunk, or hurt by storing in the fridge for too long, but now that I'm seeing signs of growth, I'm concluding there is something else wrong happening here.

My substrate since round 1 is the same, and after seeing such success the first round, can't figure out the trouble with the others.

Am I killing spores by toasting the syringe prior to injection? I'm letting out just a trickle of solution each time to cool the syringe before injecting, so really wondering what the problem is and if it has anything to do with this?

Does anyone have any idea what could be happening?

Also, if this matters, round #1 was golden teacher, round #2 costa rica, and round #3 and #4 were B+.


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"Be the change you wish to see in the world." - Gandhi


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OfflinePussyFart
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Re: Inoculation FAIL or Incubation FAIL? [Re: ShamanWarrior]
    #19241356 - 12/07/13 02:54 PM (10 years, 1 month ago)

Quote:

ShamanWarrior said:
Used alcohol to sterilize syringe between jars



No you didn't, you just sanitized it....you didn't sterilize anything.

Chemicals sanitize, heat sterilizes.

If you want to be sure you have a clean tool sterilize it with heat, to kill everything.

Alcohol does not kill all mold spores but when it does it takes time....like 10 minutes.

Wiping a flame sterilized tool with an unsterile substance like alcohol is like wiping shit on your face after a shower.

Quote:

ShamanWarrior said:
A month later, I inoculated another 12 jars with one of the syringes, same process as first round, only this time I used the flame sterilization technique on the syringe between jars. Let it cool for a few seconds and then gave the squirt. I stored these jars the same as first round, 82-85 degrees during incubation. After several weeks, these jars had ZERO sign of growth. No mycelium, no contams.



82F is too high....bring it down to 80F or below.


Jars/bags/tubs/trays should colonize @ room temperature getting ambient/indirect light.

Main pinning triggers are full colonization, FAE and Evaporation off of the substrate.

Light is a secondary pinning trigger. For tropical species temperature is not a pinning factor.

P. Cubensis are a tropical species. You could colonize at 70F and fruit at 80F with great results.

Light has been proven beneficial during all stages of mycellium growth. Mushrooms like mammals have a circadian rhythm.

You want ambient/indirect light(on a 12/12 schedule preferably) for colonization and consolidation.

You want direct/intense 6500K light on a 12/12 schedule for fruiting.

Optimal temps are mid 70s throughout the whole grow, but anywhere from 65F-80F is acceptable.

Incubation is outdated/uneeded unless temps in the range stated above cannot be kept.

The inside of the jar is always a few degrees warmer than the outside because the mycellium produces heat..mycellium tends to stall at temps above 86F , and contams thrive.

Fruiting at cooler temps tends to produce denser, meatier fruits, while fruiting at higher temps will often produce hollow, less dense stems.


Mycelium should be exposed to ambient room light from day of inoculation as has been known for many years.  Light is not a pinning trigger until after full colonization and an increase in air is given, and even then it's a secondary pinning trigger.
RR

Quote:

ShamanWarrior said:
Am I killing spores by toasting the syringe prior to injection? I'm letting out just a trickle of solution each time to cool the syringe before injecting, so really wondering what the problem is and if it has anything to do with this?



No....or flame sterilizing would not be recommended.....it is something else....

Quote:

ShamanWarrior said:
Does anyone have any idea what could be happening?



Well I think you incubated the first set too high......not sure about the rest....giving them ambient light might help the slow ones a bit.....not really sure tho....are you shaking the syringe between jars?


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OfflineShamanWarrior
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Registered: 06/26/11
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Re: Inoculation FAIL or Incubation FAIL? [Re: PussyFart]
    #19241466 - 12/07/13 03:22 PM (10 years, 1 month ago)

Hey,

Thank you for the info.

I'm going to relocate the jars to open ambient light in the room and see if that helps.

Even after 2-3 weeks of no action, are these jars still capable of producing mycelium if conditions are changed to something more suitable?

Also, finally got the heat going in my house, so temps will be kept at around 70-74 throughout the nights and days in the new location.


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"Be the change you wish to see in the world." - Gandhi


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