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Offlinepatzee
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Registered: 05/26/01
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Endospores? where, what., in rye?
    #381471 - 08/29/01 03:30 PM (15 years, 9 months ago)

Some teks recommend steeping bird seed for example (hawk tek) I vaguely remember the
reason is not only to hydrate the seed but something about possible endospores. How and why would they
be a possible contam? ( no biology book available at the moment)
I thought I also recalled that
endospores (being inside____) may not be killed by autoclaving? Does that mean that certain grains
must always be ground? I finally have all the ingredients for Alien shroom recipe, which
calls for grinding rye berries. I also wanted to experiment mixing rye berries, seeds, millet flour and such
but wanted a little more science before I approach more grain teks. Thanks everyone for reading.
Any references appreciated.



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Invisibledimitri211
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Registered: 04/27/01
Posts: 2,248
Re: Endospores? where, what., in rye? [Re: patzee]
    #381494 - 08/29/01 04:07 PM (15 years, 9 months ago)

Here you go this is an easy tek for beginners who want to try grain it is not mine it can be found at the following url with pics and come feed back.
I have never liked rye until this tek came about,
and it's works for me.
I have been told that is you let the rye sit over night in h2o then drain rinse and sterilize you can beat the icky icky's

http://www.theforestfloor.org/cgi-bin/topic.cgi?forum=12&topic=122



Ok, here it is folks, A detailed strain report on these great Puerto P. cubs. I will try to make my friend's tek as clear as possible, but feel free to ask questions, ill answer them the best I can.

What my friend did to kick the whole thing off was to germinate spores on 4 petris filled with MEA. As soon as germination was observed, transfers were made to peroxidated MEA.

These were allowed to colonize within about 1/8" of the edge of the pertis. Rhyzomorphic sectors were transfered to new plated of peroxidated MEA, and allowed to colonize the same as above.

The day before transfer to grain spawn, 1 cup of rye and 3/4 cup water was added to six jars. This was allowed to sit at room temperature, allowing all the nasties in the grain to germinate so they would be killed easily in the p. cooker. My friend recommends this procedure anytime your working with rye.

After soaking for 24 hours, plastic lids fitted with filter disks were put on the jars and tightend, then the jars lids were covered with foil in the usual manner. The jars were p. cooked @ 15 psi for 75 minutes. As soon as the jars could be handled they were removed from the cooker and shaken vigorously. These jars were allowed to cool overnite.

The next day the lids were removed from the jars and 5 ml of peroxide was added to each one, and the lid replaced. Again the jars were shaken vigorously. The jars were allowed to stand for about an hour before transfer of colonized agar.

After the hour to let the peroxide soak in, the agar dishes were opened and placed on the corner. My friend was not worried about contams because everything has peroxide in it. My friend took a scalpel and diced the agar, three plates worth, into small cubes, really tiny. Then he added a spoon of these cubes to each jar, replaced the lids and shook them again.

These jars were placed on top of the fridg and were totally colonized in 8 days, having been shaken once at about 30-40% colonized.

Now it was time for my friend to prepare worm castings. to be mixed with the colonized grain, making spawn that would eventually be mixed with pastuerized straw.

10 lb of worm castings were hydrated by misting until moist. Special care was taken not to turn the castings into mud. The castings were loaded into a foil turkey pan and baked in the oven for 2 hours at 155 degrees. The oven was turned off and the castings were allowed to cool overnite.

The next day the castings were removed from the oven and dumped into a medium sized rubbermaid container. The six jars of spawn were shaken to break up the grain and were dumped into the rubbermaid and mixed with the castings by hand. The lid was put on the container and it was put under my friend's bed til it was completely white.

When the worm castings/rye spawn was completely colonized it was time to pastuerize the oat hay. This is just basically straw with the grain heads still intact. A pillow case of this stuff was chopped into 3-4 inch pieces with tin snips then pastuerized in a water bath at 140-155 degrees for 2 hours. A brick wwas put on top of the pillow case to keep it submerged. After two hours the pillow case was removed and allowed to drain outside overnite.

The next day a 20 gallon clear rubbermaid, lid was clear also, was taped with duct tape, starting at the bottom and expending up 4 inches, to keep light from reaching the bulk substrate except on top. The straw was dumped into the rubbermaid, spread out by hand, then the tub od castings/rye was dumped on top of it and mixed in thoroughly. The lid was put on, and the container was covered with a sleeping bag til the substrate was completely colonized. When it was observed to be colonized, 1 bag of jiffy mix, unpastuerized, was dumped on top of the substrate and smoothed out by hand. It was misted til optimal moisture content was achieved. It was again covered with the sleeping bag til it looked like this.

Now came the fun part. The sleeping bag was removed and the container was fanned at least twice a day, more if my friend thought to do it.

There was no need to mist, as the straw contained enough moisture to keep the container
humid, since the lid was kept on except at fanning time.


Dr Bluethumbs Old School Toadstools
----------------------------------------------
Want great service,clean spores,fast delivery,lilshop is the place 2 be


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OfflineAnnoA
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Re: Endospores? where, what., in rye? [Re: patzee]
    #381529 - 08/29/01 05:28 PM (15 years, 9 months ago)

>How and why would they be a possible contam?
http://www.wellesley.edu/Biology/Courses/209/PowerPoint/Endospores/tsld001.htm

---------------------------------------
kickme.to/mushrooms

Edited by Anno on 08/29/01 04:30 PM.



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Offlinepatzee
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Registered: 05/26/01
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Re: Endospores? where, what., in rye? [Re: dimitri211]
    #381633 - 08/29/01 07:48 PM (15 years, 9 months ago)

Yikes Dimitri! That's beginner tek? (agar, mea? don't know squat about agar" YET",maybe I should enter
your contest) anyways.. this was helpful
ahaa!: > 1 cup of rye and 3/4 cup water was added
to six jars. This was allowed to sit at room temperature, allowing all the nasties in
the grain to germinate so they would be killed easily in the p. cooker.<

So letting them absorb the water gets them out of a vegetative state which is the potential precursor to
bacterial formation. (Thanks Anno)
I wonder if partially grinding/cracking the grain will expose them-"the nasties", even more? Or would exposing the insides make the grain protein too vulnerable to heat and therefore destroy more of the nutrients during sterlization? Or would the nutrients actually become more readily available to the mycelium? Maybe I will use same ratios & conditions, crack and soak grain, and just soak and see if there is a difference in colonization and fruiting.
Thank you Professors!

Edited by patzee on 08/29/01 06:51 PM.



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Offlinejonnyshaggs420
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Registered: 08/09/00
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Re: Endospores? where, what., in rye? [Re: patzee]
    #381696 - 08/29/01 09:23 PM (15 years, 9 months ago)

Endospores are formed by bacteria during harsh conditions in order to survive. Many can survive extremely high temperatures as well as being frozen in ice for long periods of time. Breaking the grain will not do anything to the endospores, its not that they are inside the grain but that they are resiliant. If they germinate, however, they are easy to kill.

Novelty


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InvisibleNuShroomPharmerII
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Registered: 11/03/99
Posts: 453
Post deleted by users_request [Re: patzee]
    #381746 - 08/29/01 10:40 PM (15 years, 9 months ago)



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Offlinepatzee
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Re: Endospores? where, what., in rye? [Re: NuShroomPharmerII]
    #382085 - 08/30/01 01:02 PM (15 years, 9 months ago)

NUShroom.. thanks I will try that tek for the mentally challenged.
I just got a couple pounds of rye.. will be having fun soon.

thanks everyone.. I won't mash up the rye now.




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OfflineSuntzu
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Re: Endospores? where, what., in rye? [Re: patzee]
    #382215 - 08/30/01 05:57 PM (15 years, 9 months ago)

I still think there's a great possibility that germinating endospores successfully will only lead to subsequent endospore-formation while the pressure cooker is heating up [like an SOS reaction to the slow heating-up]. I didn't read anything about how LONG it takes for Bacillus to form an endospore, that would be good information.
Just thoroughly rinsing birdseed really helps, can't speak for rye. The first wash is always murky and brown :(



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Offlinepatzee
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Re: Endospores? where, what., in rye? [Re: jonnyshaggs420]
    #382226 - 08/30/01 06:23 PM (15 years, 9 months ago)

There are spore strip indicators that are available for medical professionals
using autoclave sterilizers. Although I am quite certain that my autoclave is functioning
properly. So, perhaps some of my concerns with grains isn't a problem at all.
I'm using temps as high as 260degrees for my jars and pressures up I think around
80psi.. or it could be higher.( cant remember at the moment) That pressure should
kill em. I've run temp indicator strips inside quart jars with rings tightened and they have hit sterilization temps.Thanks everyone again. Max nutrition is my goal.




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OfflineSuntzu
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Re: Endospores? where, what., in rye? [Re: patzee]
    #382464 - 08/31/01 12:35 AM (15 years, 9 months ago)

80 psi?? Jesus, we autoclaved for scrapie, a prion disease and only went up to 30 [maybe 35] and 270 degrees.
Not saying you're wrong, but might want to check that again!



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