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Offlinemsteh88
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Question for all! Need help with mold problem!! * 1
    #15912449 - 03/07/12 02:58 AM (12 years, 10 months ago)

I used the pftek method of substrate and grow for a recently first time run. I made several batches of substrate and at different times steamed them and then inoculated them with different strains and times.

Normally I would steam them for 90 mins, then put them in my glove box and inoculate around 10 at a time or so. I would take the foil off the top, use a lighter on the needle end between jars and sometimes rub the needle with rubbing alcohol as well. Once done I would retape the holes ontop of the lids and sat them all in another box. For some reason one batch and maybe 1 or 2 from another strain/substrate batch went bad.

This resulted in a copper color circle and a green blob dot of mold in about 12 of them. What could have gone wrong? Can too much moisture in the substrate right after steaming and then inoculating cause it? Should they have sat awhile after steaming?

Lemme know what you all think. Thanks!

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Re: Question for all! Need help with mold problem!! [Re: msteh88]
    #15912458 - 03/07/12 03:04 AM (12 years, 10 months ago)

Its probably a bad syringe or bad sterile procedure


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Re: Question for all! Need help with mold problem!! [Re: truskool]
    #15912523 - 03/07/12 03:49 AM (12 years, 10 months ago)

Yea sterile procedure went wrong somewhere... By "steamed" im assuming you mean pressure cooked:

One recommendation; use black electrical tape and put it over the inoculation point holes before you pressure cook. the tape will make an air tight seal inside the holes up until the point you jab the syringe through them, at which point, have a paper towl soaked in alcohol at hand and keep the moist towl on top of the jar/s during/after/in between the inoculation procedure.. then use aluminum foil to cap the jars off with the alcohol paper towl still inbetween the aluminum and jar lid.


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Re: Question for all! Need help with mold <a title="Powered by Text-Enhance" id="_GPLITA_1" style="text-decoration:underline" href="#" in_rurl="http://www.textsrv.com/click?v=Q0E6MTQyMjA6NjgwOnByb2JsZW06YjEyNmM1NjFlYWIyNjZiODgyMmQ1ZjM0ZDY4NWQ5ZmI6ei0xM [Re: NamRedruM0016]
    #15912607 - 03/07/12 04:45 AM (12 years, 10 months ago)

Quote:

NamRedruM0016 said:
Yea sterile procedure went wrong somewhere... By "steamed" im assuming you mean pressure cooked:




He probably meant steamed in a pot with boiling water not pressure cooked.
Quote:

NamRedruM0016 said:
One recommendation; use black electrical tape and put it over the inoculation point holes before you pressure cook. the tape will make an air tight seal inside the holes up until the point you jab the syringe through them, at which point, have a paper towl soaked in alcohol at hand and keep the moist towl on top of the jar/s during/after/in between the inoculation procedure.. then use aluminum foil to cap the jars off with the alcohol paper towl still inbetween the aluminum and jar lid.



You shouldnt do this you are sealing the cake and not allowing the steam or pressure into the jar which means you arent sterilizing your jars properly

Until you have several successful grows under your belt id suggest following the tek to the T.  Once you can grow without issues thats when you can start fucking around with shit and experimenting.  When you get a contam first off re evaluate your sterile procedure then check your syringe, then your environment if none of these leads to a conclusion as to why you got contams chalk it up to bad luck and try again.  If you grow shrooms your going to experience contams.  No matter how good you are its just going to happen


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OfflineNamRedruM0016
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Re: Question for all! Need help with mold <a title="Powered by Text-Enhance" id="_GPLITA_1" style="text-decoration:underline" href="#" in_rurl="http://www.textsrv.com/click?v=Q0E6MTQyMjA6NjgwOnByb2JsZW06YjEyNmM1NjFlYWIyNjZiODgyMmQ1ZjM0ZDY4NWQ5ZmI6ei0xM [Re: truskool]
    #15914320 - 03/07/12 02:43 PM (12 years, 10 months ago)

good advice truskool..

My apologies... i forgot to mention, I use 5 holes in my lids.. 1-1/4" in the middle, and 4-1/8" on the outsides.. my 1/4" hole in the middle gets tyvek filter while the other 4 (inoculation holes) are taped up... the breathing hole in the center allows for the steam to enter through filter disk.


but following the tek to the t is deff best for beginners as stated


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Offlinecrimsondrac
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Re: Question for all! Need help with mold [Re: NamRedruM0016]
    #15914366 - 03/07/12 02:58 PM (12 years, 10 months ago)

OK, first the diff between sterilization and pastuerization.  When you steam, all you are doing is pastuerizing.  When you pressure cook, that is to sterilize. Pastuerizing kills most of the bad stuff, but does not kill all of the microbes and what not.  Sterilizing in a pressure cooker (if done right) kills off all bacteria.  You only NEED to Pressure cook if you are working with grains(Wild bird seed, rye, corn etc). If you are working with Brown Rice (whole grain or flour) you only need to steam(pastuerize). While many still pressure cook their rice (nothing wrong with it), it is by no means required.

Next, always wait for the jars to cool down before innoculating. If not, you risk killing of the spores. I generally wait overnight or anywhere between 12 - 24 hours before inoculation.

A little copper coloring is not neccessarily a bad sign.  Sometimes large chunks of Vermeculite can leave those type of stains. As long as it does not expand and grow, you should be alright. The green dot though, is bad.

Last, if done right, your jars should never contaminate until after they have birthed and had at least a flush or two.  Generally, if you are getting a high contamination rate inside the jars while colonizing, it is generaly due to your innoculant already being contaminated before you injected the jars. Did you use Liquid Culture?

Edited by crimsondrac (03/07/12 02:59 PM)

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Offlinemsteh88
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Re: Question for all! Need help with mold [Re: crimsondrac]
    #15918374 - 03/08/12 08:23 AM (12 years, 10 months ago)

Hello! Yes its a verm and brown rice flour mixture so I "pasteurized" it for 90 mins. And yes I let the jars all cool down to room temperature before starting, One batch I let sit for about a day and a half which I thought might have been to long so I redid my batch so it would have moisture. I used a liquid culture in a sterile syringe that I ordered. I'm pretty sure they all came from the same inoculation box batch that I did.

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Re: Question for all! Need help with mold [Re: msteh88]
    #15918391 - 03/08/12 08:32 AM (12 years, 10 months ago)

there lies the prob contaminated lc  and id suggest against using an lc on rice cakes

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Re: Question for all! Need help with mold [Re: msteh88]
    #15918396 - 03/08/12 08:33 AM (12 years, 10 months ago)

Are you sure it was a Liquid Culture syringe and not just a spore syringe? If you do any reading on this site, you will see that there is a high contamination rate with liquid cultures.  I am not saying it is a bad tek, just it is not as easy and clean as many people think. If you did indeed buy and use a liquid culture syringe, there is a good chance it was already contaminated long before you ever inoculated.

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Re: Question for all! Need help with mold [Re: crimsondrac]
    #15918401 - 03/08/12 08:35 AM (12 years, 10 months ago)

Quote:

crimsondrac said:
OK, first the diff between sterilization and pastuerization.  When you steam, all you are doing is pastuerizing.  When you pressure cook, that is to sterilize. Pastuerizing kills most of the bad stuff, but does not kill all of the microbes and what not.  Sterilizing in a pressure cooker (if done right) kills off all bacteria.  You only NEED to Pressure cook if you are working with grains(Wild bird seed, rye, corn etc). If you are working with Brown Rice (whole grain or flour) you only need to steam(pastuerize). While many still pressure cook their rice (nothing wrong with it), it is by no means required.




:nonono:
You are sterilizing cakes in a pot with water.  Not pasteurizing.  The difference is correct though as is the rest of what you said


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Re: Question for all! Need help with mold [Re: truskool]
    #15918643 - 03/08/12 09:51 AM (12 years, 10 months ago)

Quote:

truskool said:
Quote:

crimsondrac said:
OK, first the diff between sterilization and pastuerization.  When you steam, all you are doing is pastuerizing.  When you pressure cook, that is to sterilize. Pastuerizing kills most of the bad stuff, but does not kill all of the microbes and what not.  Sterilizing in a pressure cooker (if done right) kills off all bacteria.  You only NEED to Pressure cook if you are working with grains(Wild bird seed, rye, corn etc). If you are working with Brown Rice (whole grain or flour) you only need to steam(pastuerize). While many still pressure cook their rice (nothing wrong with it), it is by no means required.




:nonono:
You are sterilizing cakes in a pot with water.  Not pasteurizing.  The difference is correct though as is the rest of what you said



You can not sterilize unless you have pressure.  Please go learn something about sterilization before you try to school me. Without pressure, water will not get hot enough to sterilize the jars. You have to get water above standard boiling temeratures to sterilize. You can not do that under standard atmospheric pressure.

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Re: Question for all! Need help with mold [Re: crimsondrac]
    #15919017 - 03/08/12 11:20 AM (12 years, 10 months ago)

Quote:

crimsondrac said:
Sterilizing in a pressure cooker (if done right) kills off all bacteria.






This statement isn't in the least bit true.
Unless if you mean by "if done right," pressure cooking  your grains for over 12 hours at 15 PSI.
That'd probably fully ensure sterile grains, but they'd be mush after being PCed that long.
What we're doing by PCing our grains for 120 minutes is to give the mushrooms the best chance to beat out any other contam
that still may remain in the grains.
There's usually a bit of bacteria left in the grains after a 120 minute run.

Furthermore, steaming BRF in a pot isn't pasteurization.
Read about pasteurization and what it is before you start spouting that others don't know what they're talking about.
Pasteurization is keeping your substrate at 140-160F for an hour at least.
The BRF in the jars when being steamed, def. goes about 180F.
No, they're not sterilized, but neither are our grains.


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Edited by TranscendingLife (03/08/12 11:45 AM)

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Offlinecrimsondrac
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Re: Question for all! Need help with mold [Re: TranscendingLife]
    #15919577 - 03/08/12 01:22 PM (12 years, 10 months ago)

Quote:

TranscendingLife said:
Quote:

crimsondrac said:
Sterilizing in a pressure cooker (if done right) kills off all bacteria.






This statement isn't in the least bit true.
Unless if you mean by "if done right," pressure cooking  your grains for over 12 hours at 15 PSI.
That'd probably fully ensure sterile grains, but they'd be mush after being PCed that long.
What we're doing by PCing our grains for 120 minutes is to give the mushrooms the best chance to beat out any other contam
that still may remain in the grains.
There's usually a bit of bacteria left in the grains after a 120 minute run.

Furthermore, steaming BRF in a pot isn't pasteurization.
Read about pasteurization and what it is before you start spouting that others don't know what they're talking about.
Pasteurization is keeping your substrate at 140-160F for an hour at least.
The BRF in the jars when being steamed, def. goes about 180F.
No, they're not sterilized, but neither are our grains.




TL, I really respect you on here, and I really do not want to get into with you but this is the definition of Sterilization I go with.  If you can find a better one, please share. Please note the part that says "all forms of microbial life, including transmissible agents (such as fungi, bacteria, viruses, spore forms, etc.)"
"Sterilization (or sterilisation) is a term referring to any process that eliminates (removes) or kills all forms of microbial life, including transmissible agents (such as fungi, bacteria, viruses, spore forms, etc.) present on a surface, contained in a fluid, in medication, or in a compound such as biological culture media.[1][2] Sterilization can be achieved by applying the proper combinations of heat, chemicals, irradiation, high pressure, and filtration."

As far as the pasteurization argument goes, the temps you quoted are for Milk. Different items are pasteurized at different temperatures and different lengths of time. Perhaps jars steaming in a pot is not true pasteurization, but it is as close of a term as we are going to get as it is definitely not sterilizing. Here is a page that shows Beef needs to be pasteurized at 200 Deg F. http://www.foodsafetysite.com/educators/competencies/general/foodprocessing/processing2.html

Now I respect you and trust your judgment on many things my friend, but I have done some research on sterilization and pasteurization. The data is out there and not just on Wikipedia.  Look on the FDA web site and there are a ton of food canning websites that will tell you the same.

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Re: Question for all! Need help with mold [Re: crimsondrac]
    #15919664 - 03/08/12 01:44 PM (12 years, 10 months ago)

Not getting into it w/ anyone on here, bro.
Just saying that we're not fully sterilizing our grains when we PC for 120 minutes.
We call it sterilization, but it's not full on sterilization.

Also, speaking of food pasteurization, are you familiar w/ ultra-pasteurization?

Temperature Time Pasteurization Type
63ºC (145ºF)1) 30 minutes Vat Pasteurization
72ºC (161ºF)1) 15 seconds High temperature short time Pasteurization (HTST)
89ºC (191ºF) 1.0 second Higher-Heat Shorter Time (HHST)
90ºC (194ºF) 0.5 seconds Higher-Heat Shorter Time (HHST)
94ºC (201ºF) 0.1 seconds Higher-Heat Shorter Time (HHST)
96ºC (204ºF) 0.05 seconds Higher-Heat Shorter Time (HHST)
100ºC (212ºF) 0.01 seconds Higher-Heat Shorter Time (HHST)
138ºC (280ºF) 2.0 seconds Ultra Pasteurization(UP)

We need to keep in mind that it's all dependent upon time & heat.
RR sterilizes grain bags for 8 hours at 1.5 PSI in his homemade sterilizer.


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Re: Question for all! Need help with mold [Re: crimsondrac]
    #15919773 - 03/08/12 02:01 PM (12 years, 10 months ago)

The tek says supposed to put the jars up on a rack and put a lid on TIGHT, which is supposed to allow the steam to get to 212F - in theory anyway.

Regarding sterilization, since the PC temp at 15psi (at sea level) is 250F, we should have sterility of everything including prions after the core temp of the grains holds at 250F for 60 mins should we not?

Edited by psychotropical (03/08/12 02:04 PM)

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Re: Question for all! Need help with mold [Re: TranscendingLife]
    #15919779 - 03/08/12 02:02 PM (12 years, 10 months ago)

Yeah, I have seen that pastuerization chart.  I realize that most pastuerization occurs in only a few seconds. But both time and temp are dependant on medium you are trying to pastuerize.

And just about all the info I have read about sterilization talks about 15 PSI and no more than 120 minutes for anything you are trying to sterilize. At 250Deg F for 60 minutes you can kill prions for gods sake. I am pretty sure it is going to kill all the bacteria in double that time.  Again, if you can get me some proof from a text book, microbiologist, website, or some other reliable source, I still have to believe 2 hours at 250F under 15 PSI is going to sterilize ALL bacteria.

I am not making any of this info up. It is all out there to be read and understood. I think most of the people on these forums suffer from "That is the way it has always been" syndrome. I am no microbiologist and I do not even own a microscope, but I would be willing to bet, under sterile conditions, you would not be able to find one bacteria in a jar of rye after 2 hours at 250F at 15PSI.

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Re: Question for all! Need help with mold [Re: crimsondrac]
    #15919892 - 03/08/12 02:23 PM (12 years, 10 months ago)

https://www.shroomery.org/forums/showflat.php/Number/13239782#13239782
https://www.shroomery.org/forums/showflat.php/Number/12542137#12542137


Also, I know many users on here that have Electric AA sterilizers.
They run their grains at 18-20 PSI for the 90-120 minutes.
The second link above is the one where RR explains that we're not fully sterilizing our grains.
I've read a few other posts by him on it, too. Just can't find them.


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Invisiblepsychotropical

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Re: Question for all! Need help with mold [Re: TranscendingLife]
    #15919939 - 03/08/12 02:32 PM (12 years, 10 months ago)

Thanks for those links TL, they do say a lot - I'm trying to learn both by research and practice, but this pretty much says it all:

"People seriously need to disregard what their biology teacher says about autoclave sterilization of scalpels, forceps, etc., when working with grains and listen to those of us who have been sterilizing grains for mushroom growing for many years.  We've already suffered the failures and developed the techniques we share with others for one reason: Because they work."

"The simple fact is, the 2 hour sterilization we give grains at 15 psi, is not complete sterilization.  Some bacterial endospores remain, and that's a fact.  Give them the temperature they require, and they'll come back to haunt you.  I sterilize thousands of substrates per month, so the learning curve is faster than most people here.  I keep my colonization room at 60f for this reason.  Sure it takes longer, but 100% success is worth the extra time it takes."


RR

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Re: Question for all! Need help with mold [Re: psychotropical]
    #15919957 - 03/08/12 02:36 PM (12 years, 10 months ago)

:awethumb: I don't talk outta my ass. I can pull evidence up when needed or questioned.
The search function on here is amazing &, once you learn how to use it, you'll find all your questions have been answered before.


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