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bigfootscreepyuncl
Stranger


Registered: 11/15/20
Posts: 3,717
Loc: Gamehenge
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28056096 - 11/17/22 12:42 PM (1 year, 2 months ago) |
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Ah, thank you for that clarification. Either way I'm excited for those results. If it works for a liter it stands to reason that it will work for a pint
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DharmaForKarma
Tub monkey


Registered: 04/24/20
Posts: 565
Last seen: 1 day, 5 hours
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Darn it. I put a shiny new Presto in my cart. Now that itβs confirmed that the IP sterilizes grains I donβt need it.
Thank you P9. Your work matters.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: DharmaForKarma] 1
#28056171 - 11/17/22 01:22 PM (1 year, 2 months ago) |
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I'd still get the presto, it's a valuable piece of kit that every cultivator should own.
Quote:
bigfootscreepyuncl said: Ah, thank you for that clarification. Either way I'm excited for those results. If it works for a liter it stands to reason that it will work for a pint
Precisely, if it will run a L jar the pint will surely be covered.
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hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,820
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 2
#28056197 - 11/17/22 01:33 PM (1 year, 2 months ago) |
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oh yeah man. even for just doing small scale personal cultivation of actives itβs so killer. when i started i had a 6qt presto & doing the 4 pints was more than enough for a few ounces out of some shoeboxes. if youβre not trying to supply a small country with mushrooms you end up with more than enough. plus the ability to run agar & LC through it is huge. great for g2g pints as well like you said
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schpat
psychesomadelic



Registered: 09/26/21
Posts: 1,039
Loc: South Africa
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Re: Autoclave Validation and Load/Cycle Verification [Re: hazyhorse] 1
#28056228 - 11/17/22 01:47 PM (1 year, 2 months ago) |
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Awesome work
From experience with the IP you don't need the trivot. Also 6l version can hold 3 narrow mouth quart jars and the 8l version can hold 3 wide mouth jars.
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DharmaForKarma
Tub monkey


Registered: 04/24/20
Posts: 565
Last seen: 1 day, 5 hours
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Re: Autoclave Validation and Load/Cycle Verification [Re: schpat] 1
#28056241 - 11/17/22 01:59 PM (1 year, 2 months ago) |
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I run 4 wide mouth quart jars upright in my 8 quart IP. Itβs a perfect fit.
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offworld
Lurker


Registered: 08/07/21
Posts: 71
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Re: Autoclave Validation and Load/Cycle Verification [Re: schpat]
#28057223 - 11/17/22 09:42 PM (1 year, 2 months ago) |
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Quote:
schpat said: Awesome work
From experience with the IP you don't need the trivot. Also 6l version can hold 3 narrow mouth quart jars and the 8l version can hold 3 wide mouth jars.
I ran 3 regular (narrow) mouth quarts for a long time but found I could fit 4 in my 6l IP if I lined up the protrusion on the underside of the lid in the gap between jars. Just rotate the tub until the lid can engage.
I also use a silicon trivet that has a low profile, for reasons I'm no longer sure. Glad to hear I can probably ditch it.
...but now I'm questioning if I have the 6l or the 8l, going to have to double check that.
Edited by offworld (11/17/22 09:52 PM)
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UkaUka
Cpt



Registered: 05/12/22
Posts: 28
Loc: Eastern Europe
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Re: Autoclave Validation and Load/Cycle Verification [Re: offworld]
#28057735 - 11/18/22 08:06 AM (1 year, 2 months ago) |
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That's legendary. Cheers for all the effort and money you've put into this. I would just love some clarification because honestly I am still quite lost, does the time required for sterilization increase as the volume of grains increases? I got a PC designed to work at 11 PSI and I keep pirate pushing her rocket down, only done agar and pf, a fair success but I am worried about grains which is my next step. If I can run her at the designed psi would be an ease of mind. Thanks!
-------------------- Writing is good, thinking is better. Cleverness is good, patience is better. looking for any give-away spore prints.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: offworld] 4
#28057758 - 11/18/22 08:16 AM (1 year, 2 months ago) |
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Right so I have some pretty interesting preliminary observations that have my gears turning.

On the left you'll notice that the integral chemical indicator strip is black, this is the cycled ampoule, the one on the right with the blue strip is the control.
I have been keeping a close eye on the control amps to determine how long it takes to observe colour change from purple to yellow which indicates the presence of bacteria. This picture was taken at the 5 hour mark.
I know that the incubator is functioning, this is why a control is required every cycle to ensure that the incubator is actually capable of reactivating the endospores.

This picture was taken this morning at the 14 hour mark, the control amp has already been thoroughly colonized by bacteria, which means that if the cycled amp was not successfully sterilized it should have began to turn by now, there's still time however.
So, every cycle and every load so far has passed the verification process. What does this tell me.
Inferences can be made by the results regarding missing data.
If we can be assured that endospores are destroyed regardless of grain selection, size, etc, then we know that there are other factors to consider.
It appears as though our pressure cookers easily penetrate the common loads and thoroughly defeat thermophilic endospores, which means that if we have any remaining issues with grains post cycle (so long as the cycle was performed correctly) those issues will not be due to reactivated endospores. The chance of single CFU survival is 10-6
What then remains is user error and or blis compounds or some other similar product produced by either the thermal death of bacteria/other microorganisms, and/or chemical compounds from the agro industry including mycotoxins.
So yeah, there are likely to be other variables that aren't factored into our sterilization process that aren't easily solved by simple heat exposure.
Perhaps it's the case that relatively short exposures are effective against endospores while longer exposures serve to breakdown chemicals, toxins, or other inhibitory compounds.
Our grain jars contain a microscopic world of complex ineractions and organisms completely alien to our preconceptions.
I have a feeling that the 90 minute cycle for the IP will pass verification, I'm also willing to bet that the 30 minute reduction to one hour will still be sufficient for the presto.
The jury is still out so I won't jump to conclusions just yet, but the fact that all cycles have passed verification....
Very interesting.
Edited by Stipe-n Cap (11/18/22 12:07 PM)
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SirPsycho
Purple Belt in Google-Fu



Registered: 01/01/20
Posts: 6,901
Loc: Rent free in your head
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 2
#28057777 - 11/18/22 08:23 AM (1 year, 2 months ago) |
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There's two kinds of people in this world, those that draw conclusions from incomplete data sets.
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Ask me about free Ps tampanesis, Ps subtropicalis and Ps cubensis (ESS) prints Balance in life is like running on ice.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: UkaUka]
#28057778 - 11/18/22 08:24 AM (1 year, 2 months ago) |
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Quote:
UkaUka said: does the time required for sterilization increase as the volume of grains increases? I got a PC designed to work at 11 PSI and I keep pirate pushing her rocket down, only done agar and pf, a fair success but I am worried about grains which is my next step. If I can run her at the designed psi would be an ease of mind. Thanks!
If you're running standard grain jars it would appear as though you're in the clear and should be fine, the IP runs well below 15psi.
As far as volume of grain is concerned that is still yet to be determined but I would imagine that the volume of the vessel will play a role when considering cycle duration, like I said this is yet to be determined.
I will find out once I start testing spawn bags.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: SirPsycho] 1
#28057783 - 11/18/22 08:25 AM (1 year, 2 months ago) |
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Quote:
SirPsycho said: There's two kinds of people in this world, those that draw conclusions from incomplete data sets. 
Well that's not quite what I'm doing, as I've already run 5 out of 6 tests, 5 have passed and the 6th is looking promising.
Even if the 6th fails the observation remains unchanged.
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rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
Posts: 6,767
Loc: In the Gills
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28057795 - 11/18/22 08:30 AM (1 year, 2 months ago) |
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I think the volume of grain just means you may need a longer vent time if you're seeing bacteria. I would recommend trying different vent times with your max load of grains. The more evenly hot and more vented the load is the better your cycle will be. Leaving jars blank after the cycle testing different vent times shaking the blank jars every 2 weeks should get you dialed in perfectly.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: rumfor69] 1
#28057804 - 11/18/22 08:36 AM (1 year, 2 months ago) |
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Definitely.
I think that user error due to gaps in understanding are mostly to blame. When cycled correctly, and by correctly I mean attention has been payed to proper grain preparation/hydration, ample vent time including ample time to heat surfaces to working temperatures will result in sterile grains.
Jars have lot's of gaps between them which allow for the free movement of steam, packing dense bags into a presto will likely require longer vent times and longer cycle exposures to penetrate the load.
This set of tests will be coming soon, I promise ; )
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UkaUka
Cpt



Registered: 05/12/22
Posts: 28
Loc: Eastern Europe
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28057826 - 11/18/22 08:55 AM (1 year, 2 months ago) |
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That's some quick replies right there. Thank you kind sir(s).
-------------------- Writing is good, thinking is better. Cleverness is good, patience is better. looking for any give-away spore prints.
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SirPsycho
Purple Belt in Google-Fu



Registered: 01/01/20
Posts: 6,901
Loc: Rent free in your head
Last seen: 2 hours, 49 minutes
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28057861 - 11/18/22 09:18 AM (1 year, 2 months ago) |
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Quote:
p9hu7 said:
Quote:
SirPsycho said: There's two kinds of people in this world, those that draw conclusions from incomplete data sets. 
Well that's not quite what I'm doing, as I've already run 5 out of 6 tests, 5 have passed and the 6th is looking promising.
Even if the 6th fails the observation remains unchanged.
Then you must be the second kind. I don't know, I don't have a complete data set
--------------------
Ask me about free Ps tampanesis, Ps subtropicalis and Ps cubensis (ESS) prints Balance in life is like running on ice.
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bakedbeings
orbiter of truth


Registered: 09/01/20
Posts: 4,218
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28057918 - 11/18/22 09:39 AM (1 year, 2 months ago) |
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Quote:

This picture was taken this morning at the 14 hour mark, the control amp has already been thoroughly colonized by bacteria, which means that if the cycled amp was not successfully sterilized it should have began to turn by now, there's still time however.
interesting...how long are you keeping these around? and is there certainty that its long enough to rule out even a single surviving bacterium?
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ReverendMyc

Registered: 03/29/19
Posts: 1,494
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 2
#28057948 - 11/18/22 09:46 AM (1 year, 2 months ago) |
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First, I just want to get on this thread. Been following but want to keep up.
Second, P9, you are the man. I really appreciate good rigor and admire anyone who can change an opinion in the face of evidence.
Third, I trained in nuclear propulsion and operated all steam powered engines for years. That may not seem relevant, but everything I learned in Heat Transfer and Fluid Flow, which is the foundation of most of the equipment in a ship's engine room, confirms what you are finding and contradicts much of the common PC wisdom on the forum. True, it is not about sterilization for steam powered machines. But the thermo-goddamics still apply.
Glad to see what you are doing here and the way you are going about it. The world needs more proper science and people who believe in it.
-------------------- LAGM 2.024Stoned Gummys tek (Gummies from sclerotia or mushrooms) *Not just for stones any moreHow to succeed in mycology (and life) - know nothing, read everything, try something, and accept advice. Don't Panic   
Edited by ReverendMyc (11/18/22 09:50 AM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: bakedbeings] 1
#28057972 - 11/18/22 09:57 AM (1 year, 2 months ago) |
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I get rid of them after the 24 hour incubation period.
Yes the process is designed to ensure accuracy, these products are used to save lives. The likelihood of 1 CFU survival is 10-6.
@ReverendMyc, thanks.
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ruawakeyet


Registered: 05/03/21
Posts: 1,862
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 2
#28058005 - 11/18/22 10:15 AM (1 year, 2 months ago) |
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I know others have mentioned this, but at some point I would like to donate a little to this project. I don't have much, but if everyone pitched in what they could afford, a portion of the expenses could be repaid to P9.
Would you care to let us know how much coin you've dropped on these experiments so far? I'm guessing it's well over a grand already, but that might be a low guess. $2K? (Obviously you don't have to if you don't want to.)
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