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SirPsycho
Purple Belt in Google-Fu



Registered: 01/01/20
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28053532 - 11/15/22 07:54 PM (1 year, 2 months ago) |
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Rusty2096
rah rah raw in Lady gaga



Registered: 08/23/22
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 2
#28053539 - 11/15/22 07:56 PM (1 year, 2 months ago) |
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Very interesting.
Thanks for all your hard work and financial investments. I mean that
Are you going to re-attempt with the IP and decrease the sterilization time?
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!. We don't own things - things own us. Semi-solid liquid culture (SSLC)
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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: Rusty2096] 4
#28053556 - 11/15/22 08:01 PM (1 year, 2 months ago) |
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I'll reduce the cycle for the IP if it passes, I'll run reduced cycles for the IP and presto to find the minimum effective cycle. If the IP fails then there won't be any need for cycle reduction, obviously.
Time will tell, the biological indicator has the final word.
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bigfootscreepyuncl
Stranger


Registered: 11/15/20
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 1
#28053713 - 11/15/22 09:24 PM (1 year, 2 months ago) |
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Well, fuck..I just told someone a day or two ago they couldn't use a IP to sterilize grains and they said they do it regularly Can't find the thread now so if they see this I'll say 'Sorry!'. Very interesting. I wonder if there are different 'max pressures' across a variety of models Either way, you're doing Gods works here p9! This is an invaluable experiment. Thanks again
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I 5318008 NOT a virgin!
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DharmaForKarma
Tub monkey


Registered: 04/24/20
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Bigfoot, I am one of those IP users. Exclusively oats and it works. LC goes in to the jars and fruits come out of the tubs. I have never lost an inoculated jar to contamination. I have even left cooked jars on deck for many (many!) months with no visible contamination.
And yet, no matter how I tweak the variables, my jars just donβt colonize as beautifully as we often see here. My yields are not quite as high. P9βs opus on identifying contamination was illuminating for me and it pointed in one direction: Are my oats hosting bacteria that impede (but donβt prohibit) mycelial development?
Very appreciative of the work here. Looking forward to the results.
Edited by DharmaForKarma (11/15/22 09:44 PM)
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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
#28053775 - 11/15/22 10:17 PM (1 year, 2 months ago) |
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I've argued against the use of instant pots for grains many times, if this IP passes the autoclave validation protocol then I will go where the evidence leads me, until then the jury is still out with 20 hours remaining until the BI ampoule has been sufficiently incubated.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,808
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28054149 - 11/16/22 08:51 AM (1 year, 2 months ago) |
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DERRAYLD
Constructus


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Re: Autoclave Validation and Load/Cycle Verification [Re: Pastywhyte] 1
#28054152 - 11/16/22 08:54 AM (1 year, 2 months ago) |
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This is some fine work.
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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: DERRAYLD]
#28054199 - 11/16/22 09:20 AM (1 year, 2 months ago) |
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maybe this is a dumb question but is it certain that the ampoule is a good analog for large grain like an oat? or is there a possibility that the interior of a grain is more insulated? i ask because i have had to increase my venting and cook times in order to get acceptable spawn. or course altitude could be my problem
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,808
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: bakedbeings] 4
#28054239 - 11/16/22 09:55 AM (1 year, 2 months ago) |
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No one vents long enough. If youβre not venting right now itβs not been long enough.
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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] 2
#28054258 - 11/16/22 10:07 AM (1 year, 2 months ago) |
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Altitude is a consideration for sure.
Yes the ampoule is the perfect device for load verification, it's designed for that exact purpose.
Think of it like this, the center of the grain jar will be the zone of maximum insulation from heat penetration; the ampoule exterior is plastic, the interior is glass filled with endospore solution. If the ampoule is sterilized when exposed to x cycle then any grain at the center or any other location of the load would also be sterile because the ampoule is insulated by the mass of grains surrounding it and the body of the amp itself. The thermal energy will have to not only penetrate the grain mass but also the ampoule which also implies that the trapped gasses have been purged to allow for penetration of the load. If endospores are destroyed in the central mass of the load they will be destroyed throughout.
The biological indicator ampoule is the benchmark verification process for medical and pharmaceutical sterility assurance, lives depend on it.
This is why I've spent this much time and money on this, it will render an indisputable result either way.
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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] 3
#28054273 - 11/16/22 10:16 AM (1 year, 2 months ago) |
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dope. i had to ask 
its fascinating that you are the first one to actually check this. i guess i assumed this kind of experiment had been done long ago
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,808
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: bakedbeings] 1
#28054275 - 11/16/22 10:21 AM (1 year, 2 months ago) |
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Still a pretty young discipline all things considered
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SirPsycho
Purple Belt in Google-Fu



Registered: 01/01/20
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap]
#28054276 - 11/16/22 10:21 AM (1 year, 2 months ago) |
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Quote:
Pastywhyte said: No one vents long enough. If youβre not venting right now itβs not been long enough.
If you still have water in your PC, you haven't vented long enough.
Quote:
p9hu7 said:
This is why I've spent this much time and money on this, it will render an indisputable result either way.
We all appreciate your time, effort and expense.
--------------------
Ask me about free Ps tampanesis, Ps subtropicalis and Ps cubensis (ESS) prints Balance in life is like running on ice.
π
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    "Mist your balls and fan your asshole" - Pandaskis, 2023
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rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
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Re: Autoclave Validation and Load/Cycle Verification [Re: Pastywhyte]
#28054286 - 11/16/22 10:28 AM (1 year, 2 months ago) |
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Quote:
Pastywhyte said: No one vents long enough. If youβre not venting right now itβs not been long enough.
I agree with this completely
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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]
#28054301 - 11/16/22 10:41 AM (1 year, 2 months ago) |
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More is more. Thermophilic microorganisms though Thermophilic have limitations to their resistance.
Longer venting ensures trapped gasses are purged and that surfaces have ample time to evenly heat, uneven heating leads to "cold" spots, cold relative to working temperature.
Longer cycle exposure ensures that microorganisms have had sufficient exposure to heat and serve as a redundancy failsafe for some inadvertent failure somewhere in the process.
Running temperature/pressure in excess of 15psi or 121Β°C adds another layer of redundancy.
Redundancy measures are integral to successful planning, failsafe measures are a cheap insurance policy. This is why many of us always recommend more than the standard 90 minute cycle at 15psi with a 10 minute purge.
More is more. (Within reason)
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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]
#28054308 - 11/16/22 10:50 AM (1 year, 2 months ago) |
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from time to time i run into (usually older) teks that mention "not overcooking" things. i always ignored this and have run LC, agar, and brf in the same PC with my grain jars and never perceived any detrimental effect. i assume by saying more is more you are confirming that the danger of overcooking things isnt real? (within reason )
ps. it wasnt a warning about running the pot dry i mean overcooking the actual material
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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
#28054323 - 11/16/22 11:06 AM (1 year, 2 months ago) |
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Caramelized liquid media isn't something I've ever experienced and have serious doubts regarding tales of this effect.
Overcooked grains will turn to mush, this is more of an issue for grains sterilized with atmospheric steamers due to the long exposure times (24 hours or so).
I run my 75x for 4 hours after a 30 minute vent, all in my grains are exposed to very high heat for a pretty long period of time and still require cooling once removed from the unit the following day so I'd say that overcooked grains are pretty difficult to achieve without running out of water in your pc or sterilizer.
So yeah I basically made that "within reason" statement for anyone about to ask if they can vent for 2 hours and then cycle for 6, or something equally deserving of an old fashioned face palm 
I mean, if that person is out there I'm sure you'd run your presto dry long before, but lest us know if you do make that attempt
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,808
Loc: Canada
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Re: Autoclave Validation and Load/Cycle Verification [Re: Stipe-n Cap] 1
#28054339 - 11/16/22 11:14 AM (1 year, 2 months ago) |
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Quote:
p9hu7 said: More is more. (Within reason)
Couldnβt have said it better myself
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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: Pastywhyte] 5
#28054590 - 11/16/22 02:33 PM (1 year, 2 months ago) |
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20.5 hours in and the IP amp remains sterile, the control amp has incubated bacteria:

The same for the oat amps.
Surprised by the IP results so far, I'd be further surprised if it failed by the 24 hour mark.
I think I'll try the IP with the standard 90 min cycle next to see how it goes.
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