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Invisiblewind_wraith
veteran
Registered: 05/16/01
Posts: 1,200
bacteria
    #318775 - 05/16/01 06:00 PM (23 years, 6 months ago)

Hi shroomers

This is my first post here; I'm very new to the art of shroom farming (I ordered my first spore syringe about a month ago) and haven't been having much luck so far. I've started four different mycelium cultures now and have lost them all (except for my latest attempt, hopefully) to bacteria. The first time it happened I thought it was because the "vermiculite" I was using was all wrong (resulting in a lumpy, gooey sort of substrate), but my second culture, using proper vermiculite bought from a garden store, went sour as well. The third time I tried I obsessed about sterility, but to no avail -- somehow a bacterium always manages to infect my substrate, turning it sour before I can even see any mycelium growth. I've been using the standard PF Tek method with brown rice flour, and following the instructions exactly. Now I'm exploring alternative methods because, try as I might, I just can't seem to get around the bacteria problem. Any advice?



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OfflineSuntzu
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Registered: 10/14/99
Posts: 1,396
Last seen: 4 months, 11 days
Re: bacteria [Re: wind_wraith]
    #318802 - 05/16/01 06:33 PM (23 years, 6 months ago)

Bacteria are likely the result of incomplete sterilization, though it is possible to have it 'wicked' from the outside if there is nutrient connecting inside and outside--make sure the jars are wiped clean.
Be sure to sterilize as close to 15 PSI as possible, for a hour or a bit more. Make sure your pressure cooker has built up a sufficient head. Aunt Jemima lets hers vent until the pressure needle just starts to move.
If it were mold, I'd say to be careful of how, where you cool your cooker. Since it's bacteria, and assuming your inoculum is clean [implement a negative control, an uninoculated jar that is incubated with the others] this should cover the vectors as far as I can tell.
Try, try again,and welcome to the contaminant club.

The above information is complete fiction.

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Invisiblewind_wraith
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Registered: 05/16/01
Posts: 1,200
Re: bacteria [Re: Suntzu]
    #319021 - 05/16/01 11:48 PM (23 years, 6 months ago)

Well, of course I don't have a pressure cooker (yet?), maybe simply boiling my jars in a pot isn't enough. I'm going to try the control experiment you mentioned just to make sure it isn't my spore syringes which are contaminated, though. On the subject of alternative substrate materials, has anyone tried boiled grass and leaves? I got this idea from one of the grow methods in the Grow/Find section, and it occurred to me that this might be closer in composition to Cubensis' favorite food, cow shit (= partially digested grass). So I cooked up a big pot of grass clippings and dead leaves and used that as a substrate -- I'm hoping it might be a little less vulnerable to the bacteria which have been ruining all of my brown rice cultures.



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I am a figment of my own imagination
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OfflineTaz
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Registered: 03/18/01
Posts: 1,090
Last seen: 23 years, 2 months
Re: bacteria [Re: wind_wraith]
    #319132 - 05/17/01 04:02 AM (23 years, 6 months ago)

ok, first of all U need to make sure that when you boil your jars that the lids are on loose and that you cover the pot and boil for 1 hour, then when it's cool enough to handle, tighten lids and take them out of the pot and place the jars somewhere out of direct sunlight and leave them there for a couple of days, if no strange colors apear then it's time to inject the spores, don't touch the needle are let it touch anything else, if it does then flame sterilize the needle. since your haveing such a problem I would flame sterilize inbetween each jar, (the needle) and then keep the lids on tight and make sure the needle hours are taped up, It may also be a good ideal to wear surgical gloves while injecting spores. only use 1 or 2cc per jar.

also another tip for U to try...when you make up the substrate with BRF/verm, wash all surfaces that your working on really good, then wipe the counter with 91% rubbing alcohol. then place a clean perferably fresh laudry dry towel and work on top of that. wash all utensiuls in really hot water using dish soap, i.e jars, lids spoons mixing bowls, wash all measuring cups to. before you do all this make sure you wash your hands and arms really good with as hot of water as U can. ok try this.

the leaves and grass your talking about I wouldn't do that at all, that is not a good substrate to use. another thing to keep in mind when injecting spores is to make sure your doing it in a draft free area, turn off heater/ac if air is moving around so is all the stuff that will contaminate your jars, it will get on the needle and will then be injected right in the cakes as well, always make sure when U do make up substarte and put it in jars that you wipe the 1/2 inch from the top with tissue and fill up with dry verm. hope this helps.

"Out of chaos...comes order...." AND THE FSR..


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Invisiblewind_wraith
veteran
Registered: 05/16/01
Posts: 1,200
Re: bacteria [Re: Taz]
    #320333 - 05/18/01 05:20 PM (23 years, 6 months ago)

Mmkay. I've now inoculated a new jar using rye flour and prepared an identical, non-inoculated second jar to see whether it might be my syringe which is contaminated. No sign of growth on the grass/leaves jars yet, but I opened one up and it smells alright, so at least I've eliminated the bacteria problem. Now it's just a question of whether my spores are going to germinate on this stuff.



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I am a figment of my own imagination
http://go.to/FreeSporeRing

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