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OfflineteladiS
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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25873939 - 03/14/19 03:42 PM (2 months, 9 days ago)

Quote:

Pinback said:
Food dating is for food quality, not safety (see here )





I don't refer to food dating of best before dates, but expiration dates found on many, many canned goods. Expiration dates are a thing, but don't seem to be a thing in your country.

Quote:

Pinback said:

No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.




I don't think you are understand what I am saying. Please re-read the sentence:

Quote:

It's also why you'll never ever find any sterilization method promising 100% sterilization rate. It's just not doable.




But you seem to be contradicting yourself. You first say, "I have only ever seen that notion here" referring to stareatclouds saying you can't sterilize completely, and then say, "will reduce the risk of contamination of a given item to some acceptable low number.". Not sure if you realize this.

Eh, missed an entire conversation in between, sorry guys for throwing the flow out.


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InvisiblebodhisattaM
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Re: PC'd Grain - Life Span? [Re: teladi]
    #25873941 - 03/14/19 03:44 PM (2 months, 9 days ago)

Quote:

teladi said:
Quote:

Pinback said:
Food dating is for food quality, not safety (see here )





I don't refer to food dating of best before dates, but expiration dates found on many, many canned goods. Expiration dates are a thing, but don't seem to be a thing in your country.

Quote:

Pinback said:

No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.




I don't think you are understand what I am saying. Please re-read the sentence:

Quote:

It's also why you'll never ever find any sterilization method promising 100% sterilization rate. It's just not doable.




But you seem to be contradicting yourself. You first say, "I have only ever seen that notion here" referring to stareatclouds saying you can't sterilize completely, and then say, "will reduce the risk of contamination of a given item to some acceptable low number.". Not sure if you realize this.

Eh, missed an entire conversation in between, sorry guys for throwing the flow out.



Expiration dates and best by dates are guidelines put out by the company. They really have nothing to do with actual expiration.


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OfflineteladiS
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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25873952 - 03/14/19 03:54 PM (2 months, 9 days ago)

Best by, for sure, but expiration dates are less than guidelines and have very specific definitions, in many regions. It's still dictated by the manufacturer, and based on when the manufacturer expects the goods to be unsafe for consumption.


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25873968 - 03/14/19 04:12 PM (2 months, 9 days ago)

Most expiration dates are more for the packaging then the food or contents


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25873982 - 03/14/19 04:19 PM (2 months, 9 days ago)

Quote:

teladi said:
Best by, for sure, but expiration dates are less than guidelines and have very specific definitions, in many regions. It's still dictated by the manufacturer, and based on when the manufacturer expects the goods to be unsafe for consumption.



Not really. Like cron said usually its packaging material or quality of food. Not safety.


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InvisibleMateah
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Re: PC'd Grain - Life Span? [Re: ShaperDreaming]
    #25874039 - 03/14/19 04:46 PM (2 months, 9 days ago)

[disclaimer]Just wanna say I really enjoy piecing together this puzzle together with you fellow mushroom people.
I hope my multitude of questions and statements don't cause irritation or nothing I'm just inquiring about this subject because it's interesting.[/disclaimer] :eatingout:


Alright, lets get inflammatory then..  :doggull:


It feels like I'm getting (or interpreting), somewhat inconsistent theories on the process of heat sterilizing endospores. I'm just trying to separate what we KNOW from what we 'have faith in' or 'most likely is so'.


Consensus/authorities around here hint to us that endospores are not just resilient as fuck, but you'd almost be silly to assume that 15min on 15PSI (effective heat cycle) will kill them completely. But then again I don't see anyone stressing about GWA or GWLC that's been sterilized at 15PSI for 15min :shrug: This makes me wonder why are these endospore containing nutrient rich LIQUIDS considered sterilized in 15min but the same endospores in grain (that sustain the same 'effective heat cycle') are only considered 'sufficiently incapacitated'. I have previously asked about if one can not simply just increase the pressure(heat) and the duration of the cycle to be able to effectively sterilize grain, and I was left with the impression that endospores are quite simply too tricky and too unpredictable that we'll ever be able to say that a grain jar has been effectively sterilized. And yet grain is FAR from optimal for propagating these organisms while nutrient rich liquid media's are pretty CLOSE to optimal environments for these organisms.



I get it that a grain jar possibly contains much higher spore count than GWA or GWLC and thus increases the chances of spores surviving. But this then begs the question; are there any statistics on these 'REALLY really thug life' endospores that somehow survive prolonged high temps while 99.99% of their buddies die with their mouths open already even before 15 min? (I understand that conditions can be environmental as well, like dry pockets between/inside grain) but I also know that there is no reason one shouldn't be able to always reach 'desired tepms/duration' within each grain if properly prepared and properly executed sterilization cycle.

(meaning: if your goal is to reach effective heat cycle of 15min/121C when sterilizing liquid media, then all you need to do is boil the liquid media under pressure for 15min. 15PSI makes the boiling point of water will rise to 121C. So you have permiated to every cell with 'desired temps'. But if you're sterilizing hydrated solids like grain inside qt jars then you're also dealing with partial insulation. From what I can compute so far the only practical solution would be to raise the temperature and prolong the heat cycle to compensate for the insulation. Let's say 15PSI allows for 114C to reach into the center of every grain within an hour. So you raise the pressure for example to 20-30PSI and add 30min more to the cycle and you're guaranteed that more than 114C will permeate every grain.


(I don't mean to sound so sure or cocky, I'm neither (I hope) everyone should read  my statements as mere observations/opinions/speculations, I'm just grateful for other takes on it and for clearing up facts for me if I happened to misunderstand something :takingnotes:


Okej here's another question I can't find the answer to online, is Bacillus subtilis fairly homogenous across individual organisms when it comes to heat resistence or can the differences vary greatly? Obviously weaker than average spores are no concern, but how stronger are the 'stronger than average' if they even exist? Yeah this is my question for now :bongload:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25874051 - 03/14/19 04:53 PM (2 months, 9 days ago)

Like ive said everywhere and numerous times liquids are extremely easy to sterilize. Gravity autoclaves have a hard time getting air out they fail the bowie-dick test unlike vacuum pumped autoclaves.

They make spore indicators. They're loaded with 10^7 or more endospores. They get put in the middle of a load in the middle of an autoclave. 15m at 15psi kills them to a sterility assurance level of 10^-7 chance of a single survival. But we are sterilizing grain which is completely different.


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InvisibleMateah
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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25874149 - 03/14/19 05:36 PM (2 months, 9 days ago)

Thanks for the response and so you know you're not wasting your time either lol, I have a private journal notebook where I save useful posts like your last post here, Ive read numerous of your posts about the strips since way back, very appreciative of your patience!

I understand there are different types of Bacillus organisms and different types of grain. I'm mostly asking around if anyone knows about specific numbers and shit  when it comes to the science of killing and incapacitating endospores with heat sterilization?


I've been looking online for quite a bit now, downloaded some PDF's and read also about endospore sterilization and propagation on a forum this week but I can't seem to find this specific information anywhere :shrug:


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25874266 - 03/14/19 06:38 PM (2 months, 9 days ago)

Quote:

teladi said:
But you seem to be contradicting yourself. You first say, "I have only ever seen that notion here" referring to stareatclouds saying you can't sterilize completely, and then say, "will reduce the risk of contamination of a given item to some acceptable low number.". Not sure if you realize this.





Apologies if I was not clear, let me try again. I have only ever seen here the notion that an item cannot be completely sterilized (or that it would take more than 8 or 24 hours at 121 C) to do so. Yes, there are bacteria that will survive long times at 121 C, but they are typically found in hydrothermal vents and are not relevant here.

For practical purposes, an item is sterilized for a time that is calculated or estimated to reduce the risk of contamination to an acceptable level. That level will vary depending on what is sterilized.

Suppose, as an example, that you have a 1 log reduction of some bacterium every minute at 121 C, and that there are 100 viable bacteria in every jar you want to sterilize. The bacterial count is unrealistic, but will show my point.

After 1 minute at 121 C, there are 10 viable bacteria, per jar, on average (1 log reduction).
After 2 minutes, there is 1 viable bacterium.
After 3 minutes, there is 1 bacterium in every 10th jar.
After 4 minutes, there is 1 bacterium in every 100th jar.
After 5 minutes, there is 1 bacterium in every 1000th jar. And so on. This is what the model statistically predicts, and will naturally differ in reality. For example, heating times and heat transfer has to be taken into account.

So, sterilizing 1000 jars for 5 minutes according to the example, 999 of them are sterile (again, statistically and according to the model) and 1 of them has 1 viable bacterium. This is not contradicting what I wrote earlier.


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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25874799 - 03/14/19 11:48 PM (2 months, 9 days ago)

Quote:

Pinback said:
Quote:

And also in microbiology, you program your autoclave cycle against the most common expected contaminants. Every now and then sterilization fails 'cause you have something weird creep in, and you have to throw out your entire experiment, or you sequence something completely different.

For every time period, at temperature you experience a log reduction in the number of CFUs in the thing you are sterilizing.  Basic math tells you that you are unlikely to ever reach zero with a typical CFU load, but rather you take it down to an acceptable level without overly deteriorating the substrate.

It's also why you'll never ever find any sterilization method - beit filters or whatever - promising 100% sterilization rate. It's just not doable.




No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.




This is why I said, "How is that sterilization?"

Sterilization in microbiology is killing ALL forms of life, including currently active, as well as dormant endospores. Something is either considered sterile or not. Nobody would say 95% sterile. My syringes and needles I aspirate LC into come sterile. You're saying they still have bacteria and/or mold spores on them, just at a low enough amount to be called "sterile?" Or are you just splitting hairs that we'll never truly know 100%?

Why would a filter announce a sterilization % in the first place? They don't sterilize anything, they prevent things from passing through.

I don't really understand what your argument is and seems to flip-flop back in forth, but maybe I'm just tired. I DO 2 HOURS 16-20PSI almost every time. I don't know the starting bioburden, but this works for me. :smile:


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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25874804 - 03/14/19 11:51 PM (2 months, 9 days ago)

Quote:

Mateah said:
I understand there are different types of Bacillus organisms and different types of grain. I'm mostly asking around if anyone knows about specific numbers and shit  when it comes to the science of killing and incapacitating endospores with heat sterilization?


I've been looking online for quite a bit now, downloaded some PDF's and read also about endospore sterilization and propagation on a forum this week but I can't seem to find this specific information anywhere :shrug:




Whoa! If only a handful of people had repeatedly said exactly this repeatedly throughout the thread? Come on Mateah, if modern science can't document this conclusively, your coir-water-grain-vessel contraption is needed now more than ever.


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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25874857 - 03/15/19 12:19 AM (2 months, 9 days ago)

Quote:

Pinback said:
Sure. Typically, I weigh out 180 g of rye in a glass and add 180-210 g (depends on the rye) of water at 70-100 C. I put on a lid having a polyfill plug, and aluminium foil. Then, I pressure cook at 15 psi for 60-90 min, counting from when 15 psi is reached. After the pressure has gone down, I take out the jars and shake them while still hot, then allow them to cool in a clean place, which can be the pressure cooker.

I have also done it with pre-hydrated grains, with additives like gypsum, coffee waste and vegetable oil, and in filter bags, though most of my experience is like the paragraph above. Also bigger bags with sawdust, dung and straw, but then I have kept them longer at 15 psi (like 2-4 h) to allow for heat transfer. They last at least for several weeks.

The time I have waited with grain jars is at least several months (as written earlier in the thread). Right now, I have some filter bags with rye and coffee waste sitting that I sterilized February 12th. It is not that long, but they look great, and smell great through the filter.

My usual method of inoculation is with agar wedges.




How are your rye bags?


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25874920 - 03/15/19 01:01 AM (2 months, 9 days ago)

Maybe you think my test jar 'contraption' is ridiculous but I bet that I can get still get away with attaining at least some useful data from experimenting with sterilizing liquids, grain, and other contraptions at different temperatures and durations to later inoculate the test jars with sterile water or LC or agar cultures, depending on which hypothesis I'm trying to confirm or deny..


For example bod mentioned repeatedly that liquids sterilize completely different from grain, I get that.
Of course overhydrated grains submerged in water will conduct heat more like a liquid and less like insulated dry solid.



That's why my idea for another 'contraption' test jar is to sterilize grain in one jar and GW in another one and then add the sterilized GW to the grain jar. That way I won't make the sterilization of the grain more effective but I will still give the surviving endospores on that grain a chance to colonize a nutritious liquid broth if it wants to. See? I worked around that problem cause I'm constantly thinking about this experiment and I'm motivated by curiosity.



I'm way past noob and I have been for a while, but that doest mean that I won't be asking the same question again maybe in two years If I have a two year break from growing. Life throws curve balls you know :bongload: I'm not trying to prove anyone wrong or prove anything about anyone. I can also get frustrated when my long time JJ students ask me to show them basic techniques that I already showed them a bunch of times, but then again it's not my place to question their personal dedication to the art, I'm there just as a friend/guide showing them how it's done (how I do it). But if students ask me to prepare them for a tournament then I'll be more of a general and less of a friendly guide. But out of competition It's always very relaxed setting :bongload:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25874954 - 03/15/19 01:18 AM (2 months, 9 days ago)

posting to read through later


--------------------
Everything i say is completely hypothetical...


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25875071 - 03/15/19 03:09 AM (2 months, 9 days ago)

Quote:

stareatclouds said:
Sterilization in microbiology is killing ALL forms of life, including currently active, as well as dormant endospores. Something is either considered sterile or not. Nobody would say 95% sterile. My syringes and needles I aspirate LC into come sterile. You're saying they still have bacteria and/or mold spores on them, just at a low enough amount to be called "sterile?" Or are you just splitting hairs that we'll never truly know 100%?




No, I completely agree that what we call sterile has no viable organisms. My point is that, for practical purposes, there will always be a small but acceptable risk of something being non-sterile after a sterilization procedure. Not that a sterile item itself has some small but acceptable number of bacteria. Do you see the difference?

My rye bags were subsequently inoculated, with no sign of contamination as far as I can tell.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25880450 - 03/17/19 06:01 PM (2 months, 6 days ago)

Quote:

Mateah said:
Maybe you think my test jar 'contraption' is ridiculous but I bet that I can get still get away with attaining at least some useful data from experimenting with sterilizing liquids, grain, and other contraptions at different temperatures and durations to later inoculate the test jars with sterile water or LC or agar cultures, depending on which hypothesis I'm trying to confirm or deny..


For example bod mentioned repeatedly that liquids sterilize completely different from grain, I get that.
Of course overhydrated grains submerged in water will conduct heat more like a liquid and less like insulated dry solid.



That's why my idea for another 'contraption' test jar is to sterilize grain in one jar and GW in another one and then add the sterilized GW to the grain jar. That way I won't make the sterilization of the grain more effective but I will still give the surviving endospores on that grain a chance to colonize a nutritious liquid broth if it wants to. See? I worked around that problem cause I'm constantly thinking about this experiment and I'm motivated by curiosity.



I'm way past noob and I have been for a while, but that doest mean that I won't be asking the same question again maybe in two years If I have a two year break from growing. Life throws curve balls you know :bongload: I'm not trying to prove anyone wrong or prove anything about anyone. I can also get frustrated when my long time JJ students ask me to show them basic techniques that I already showed them a bunch of times, but then again it's not my place to question their personal dedication to the art, I'm there just as a friend/guide showing them how it's done (how I do it). But if students ask me to prepare them for a tournament then I'll be more of a general and less of a friendly guide. But out of competition It's always very relaxed setting :bongload:



Have you looked into any papers from doc holiday or aloha?
They have done by far the most research and these days do a hydrate and light sterilizing then 24 hours latwr do 4 hours at 60 psi....yes 60


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InvisibleMateah
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Re: PC'd Grain - Life Span? [Re: cronicr]
    #25880491 - 03/17/19 06:25 PM (2 months, 6 days ago)

:takingnotes: cheers!

Edit: OK I tried searching 3 different doc holiday's, nothing and found no Aloha either.. But both sound familiar tho..


Edited by Mateah (03/17/19 07:09 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25881257 - 03/18/19 03:17 AM (2 months, 6 days ago)

You can read about him here, here or in comments here :smirk:


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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25881352 - 03/18/19 06:27 AM (2 months, 6 days ago)

Quote:

Pinback said:
No, I completely agree that what we call sterile has no viable organisms. My point is that, for practical purposes, there will always be a small but acceptable risk of something being non-sterile after a sterilization procedure. Not that a sterile item itself has some small but acceptable number of bacteria. Do you see the difference?

My rye bags were subsequently inoculated, with no sign of contamination as far as I can tell.




Right, and the acceptable risk is that window we have to introduce mycelium to overtake the substrate in case anything is leftover. Either way, everyone's situation is different. You should document these month old grows, for sure. Check the link in my signature.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25881366 - 03/18/19 06:55 AM (2 months, 6 days ago)

John Holliday not doc Holliday. His doctorate is honorary. I wouldn't call him a doctor.

Aloha does some weird stuff like double sterilization.


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