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InvisibleDoc_T
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Paul Stamets, Endospores, and You
    #14053216 - 03/02/11 08:09 AM (12 years, 10 months ago)

I'm here to explain endospores in the context of mushroom cultivation, and why they are not a concern.
I've said before that endospores are like bigfoots- they are real, but they won't cause you any trouble.

Here's why:

A bacterial endospore is not a spore or seed, it's an encrusted form of a bacterium.
Not all bacteria even can produce endospores, btw. It's a specialized gimmick. But some do.
The endospore forms when the live bacterium encounters dry conditions. Endosporulation is a physical response to the chemical changes which come with dehydration.
Bacteria multiply rapidly, we know this. But always the relationship with endospores is one-to-one. A bacterium becomes an endospore, it does not produce them.

Stamets mentions endospores because his grain prep method may allow some endospores to survive.
"May" and "some" are important disclaimers- but there's a much more relevant observation to be made here.

Ok then- endosporulation is a response to dehydration. What happens upon rehydration?
The endospore quickly reverts to the form it had before, a live active bacterium and gets on with business.
Endospores are tough, and some few may survive a casual sterilization.
Live active bacteria are not tough and are easily killed by the sterilization methods we use.

Here's the key point: if you soak, simmer, strain your grain, there are no endospores. None.
No endospores go into your jars, no endospores go into your PC.
There are no endospores in a jar of hydrated grain.

I'm off to work, have a good day!


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InvisibleGerman Kahuna
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Re: Paul Stamets, Endospores, and You [Re: Doc_T]
    #14053262 - 03/02/11 08:24 AM (12 years, 10 months ago)

Quote:

Doc_T said:
The endospore quickly reverts to the form it had before, a live active bacterium and gets on with business.



Define "quickly".

And I agree, especially with your last paragraph. That's why I advertise the soaking period as the safest way to make the endospores revert from dormant to active state before the grains are being pressure cooked and not after when they are suspended in your moist substrate.


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OfflineRogerRabbitM
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Re: Paul Stamets, Endospores, and You [Re: Doc_T]
    #14053344 - 03/02/11 08:51 AM (12 years, 10 months ago)

Doc, if there's something you don't understand, just ask, or do some searching, but you can't just make shit up.

Endospore forming bacteria, which comprise nearly all of the genus Bacillus, do not need to dry out to form endospores.  They can be easily observed with a cheap microscope on petri dishes.  Some can form endospores in as short a period of an hour, while others take 24 hours.  This is why long soaks cause more harm than good.  In addition, bacteria multiply every 20 minutes or so, so you can see how a long soak is going to leave one with more endospores than he started with, and there's no drying out going on in soak water.

These bacteria also don't just 'change state' back and forth, but they actually produce sporangia and free spores.

For some more reading, here's a couple links.  You can find many papers online with a simple search, or take a biology course at a local community college.

http://www.textbookofbacteriology.net/Bacillus_2.html
http://www.bioinf.uni-leipzig.de/~ilozada/papers/paper_Bacillus/Taxonomy_Genus_Bacillus.pdf


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Re: Paul Stamets, Endospores, and You [Re: German Kahuna]
    #14053345 - 03/02/11 08:51 AM (12 years, 10 months ago)

Quote:

German Kahuna said:
Define "quickly".



:thumbup:

Also 121C for 90 minutes will kill anything, including endospores (but except prions, as far as I know). If your substrate reaches those temperatures you don't need to worry about endospores (even if you do not soak your grains). And that's why you need to PC a spawn bag for 2 hours or more: more substrate=lower internal temperature.


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OfflineRogerRabbitM
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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14053407 - 03/02/11 09:04 AM (12 years, 10 months ago)

Quote:

Also 121C for 90 minutes will kill anything, including endospores




This is also not correct.
RR


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14053505 - 03/02/11 09:26 AM (12 years, 10 months ago)

Quote:

RogerRabbit said:
Quote:

Also 121C for 90 minutes will kill anything, including endospores




This is also not correct.
RR




121C for 90 minutes destroys all microorganisms, including endospores. If you are referring to extremophiles, that's another story. You won't find them in your house or in your grains. But I am always eager to learn new things from you.


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Offlinei GrOw StUFF
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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14053525 - 03/02/11 09:31 AM (12 years, 10 months ago)

Quote:

LeopardMan said:
Quote:

RogerRabbit said:
Quote:

Also 121C for 90 minutes will kill anything, including endospores




This is also not correct.
RR




121C for 90 minutes destroys all microorganisms, including endospores. If you are referring to extremophiles, that's another story. You won't find them in your house or in your grains. But I am always eager to learn new things from you.




I was under the impression that some mold and bacteria can survive very high temps for an extended period of time....They are just "kicked in the dick" and will take weeks to months to recover.....


--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.


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Re: Paul Stamets, Endospores, and You [Re: i GrOw StUFF]
    #14053577 - 03/02/11 09:42 AM (12 years, 10 months ago)

Mold? I would be very surprised. Some rare extremophilic bacteria maybe. Let's say: 121C for 90 minutes will kill anything (except prions and some extremophilc bacteria). Is that correct RR?


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Edited by LeopardMan (03/02/11 10:47 AM)


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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14053665 - 03/02/11 09:57 AM (12 years, 10 months ago)

Nope.  Some bacterial endospores will survive.  Mold will be nuked at temps below boiling. 

Grains that are properly prepared and 'sterilized' at 121 C for 20 minutes(measured at the center of the grains) will be fine for the two weeks or so it takes for the mushroom mycelium to gain the upper hand. 

Quote:

Doc_T said:

Here's the key point: if you soak, simmer, strain your grain, there are no endospores. None.
No endospores go into your jars, no endospores go into your PC.
There are no endospores in a jar of hydrated grain.





I just couldn't let this notion that endospores only form from dehydration and are gone when moistened go without challenge. It's just flat-out wrong.  Bad info like this causes many new growers to fail, and we need accurate info to help people succeed.  Otherwise, we may as well close the site down and let each person figure it all out for himself like we had to do 30 years ago when there were very few books and no internet.
RR


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14053730 - 03/02/11 10:10 AM (12 years, 10 months ago)

ok good to know...Didn't know there wasn't any "tough" mold spores....


--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.


Edited by i GrOw StUFF (03/02/11 10:11 AM)


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InvisibleLeopardMan
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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14053755 - 03/02/11 10:15 AM (12 years, 10 months ago)

I don't follow you :rolleyes: I have different information I guess. This is a well-known biology textbook. But I've found the same thing in several books and websites :shrug:


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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14053900 - 03/02/11 10:51 AM (12 years, 10 months ago)

:popcorn:


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Invisible13shroomsM
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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14054101 - 03/02/11 11:31 AM (12 years, 10 months ago)

Quote:

LeopardMan said:
I don't follow you :rolleyes: I have different information I guess. This is a well-known biology textbook. But I've found the same thing in several books and websites :shrug:





biology and mycology are 2 very different creatures. 

:nothingtoadd:


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InvisibleGerman Kahuna
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Re: Paul Stamets, Endospores, and You [Re: 13shrooms]
    #14054137 - 03/02/11 11:37 AM (12 years, 10 months ago)

Erm... since when are "biological bacteria" different from "myclogical bacteria"? I don't get it.
Mycology is still a subarea of biology as far as I am concerned.


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Re: Paul Stamets, Endospores, and You [Re: 13shrooms]
    #14054140 - 03/02/11 11:37 AM (12 years, 10 months ago)

If you want to kill all endospores you can do it with Tyndall's "Fractional Sterilisation".
But it will cost you at least 2 day's.


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Re: Paul Stamets, Endospores, and You [Re: Luger0815]
    #14054143 - 03/02/11 11:38 AM (12 years, 10 months ago)

Tyndallisation doesn't kill endospores any more effectively than proper soaking and sterilizing.


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Re: Paul Stamets, Endospores, and You [Re: German Kahuna]
    #14054157 - 03/02/11 11:41 AM (12 years, 10 months ago)

You may be right, i did not noticed any difference, in success, between these.


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Invisible13shroomsM
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Re: Paul Stamets, Endospores, and You [Re: German Kahuna]
    #14054160 - 03/02/11 11:42 AM (12 years, 10 months ago)

Quote:

German Kahuna said:
Erm... since when are "biological bacteria" different from "myclogical bacteria"? I don't get it.
Mycology is still a subarea of biology as far as I am concerned.





Im just sayin I dont go to KFC for fish or vice versa. :shrug:

Im not schooled in any type of science Im mearly sayin alot of myco related things dont always go by the books.  :2cents:


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Re: Paul Stamets, Endospores, and You [Re: German Kahuna]
    #14054161 - 03/02/11 11:42 AM (12 years, 10 months ago)

Quote:

German Kahuna said:
Mycology is still a subarea of biology as far as I am concerned.




:whathesaid: besides a bacterium is a bacterium 13 :wink:


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Re: Paul Stamets, Endospores, and You [Re: 13shrooms]
    #14054183 - 03/02/11 11:46 AM (12 years, 10 months ago)

LeapordMan, this is how i understand it: try sterilizing a grain jar and not inoculating for > 2 weeks. After the two week point eventually it will start to grow some other forms of life. If you have made proper lids and have not disturbed the jar much than its safe to assume the life has come from within. i.e. when soaking/sterilizing some microscopic life remains and eventually will make itself visible. 

Sterilizing under pressure only kills most of the microorganisms giving the cultivator a window of opportunity to grow mushroom mycelium :smile:


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Invisible13shroomsM
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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14054193 - 03/02/11 11:48 AM (12 years, 10 months ago)

Quote:

LeopardMan said:
Quote:

German Kahuna said:
Mycology is still a subarea of biology as far as I am concerned.




:whathesaid: besides a bacterium is a bacterium 13 :wink:





yeah, yeah, :dumblol:


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Re: Paul Stamets, Endospores, and You [Re: deadmandave]
    #14054216 - 03/02/11 11:54 AM (12 years, 10 months ago)

Of course given the presence of billions of germs, spores, microorganisms you will never achieve 100% sterility in your jars by PCing them for 30, 60 or even 90 minutes. It's simply a statistical equation. Even 99.999999999% sterility means that statistically only 1 in every 100 billion possible contaminants will survive, but it is still not 100%.


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Re: Paul Stamets, Endospores, and You [Re: deadmandave]
    #14054289 - 03/02/11 12:12 PM (12 years, 10 months ago)

Quote:

deadmandave said:
some microscopic life remains and eventually will make itself visible.




What kind of microscopic life form survives after 90 minutes at 121C (except those mentioned above)?

GK, I totally agree with you. If you we don't achieve 100% sterility it is because of improper (or not adequate) sterilization equipments and/or procedures (hey, we have to work in a kitchen :wink:). But in theory there are no endospores which can survive those temperatures :shrug:


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Re: Paul Stamets, Endospores, and You [Re: 13shrooms]
    #14054638 - 03/02/11 01:31 PM (12 years, 10 months ago)

Quote:

13shrooms said:
Quote:

German Kahuna said:
Erm... since when are "biological bacteria" different from "myclogical bacteria"? I don't get it.
Mycology is still a subarea of biology as far as I am concerned.





Im just sayin I dont go to KFC for fish or vice versa. :shrug:

Im not schooled in any type of science Im mearly sayin alot of myco related things dont always go by the books.  :2cents:





in my next of the woods KFC and Long John Silver are in the same building..just saying :-x

:nothingtoadd:


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Re: Paul Stamets, Endospores, and You [Re: biologys]
    #14054663 - 03/02/11 01:34 PM (12 years, 10 months ago)

Quote:

biologys said:
Quote:

13shrooms said:
Quote:

German Kahuna said:
Erm... since when are "biological bacteria" different from "myclogical bacteria"? I don't get it.
Mycology is still a subarea of biology as far as I am concerned.





Im just sayin I dont go to KFC for fish or vice verse. :shrug:

Im not schooled in any type of science Im mearly sayin alot of myco related things dont always go by the books.  :2cents:





in my next of the woods KFC and Long John Silver are in the same building..just saying :-x

:nothingtoadd:




corporations dont count :tongue2:  what are we talkin about again...


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Re: Paul Stamets, Endospores, and You [Re: LeopardMan]
    #14054736 - 03/02/11 01:44 PM (12 years, 10 months ago)

Bear in mind, the 121C for 20 minutes figure often sited in the literature is for organisms subjected directly to the steam.  A grain jar or bag isn't the same thing, and the heat doesn't travel through the loosely packed contents of a grain jar the way it would if it was a scalpel or inoculating loop sitting on a tray in the autoclave.  The center of a grain jar or spawn bag is heated by conduction, and this process takes time.

You guys can argue sterilization theory all you want, but the bottom line is that experience shows otherwise.  Those of us who used to get a lot of bacterial contamination, but now we don't, try to teach what we've learned.

However, it was the original post with bad information stated as absolute fact that got sand in my panties earlier.  It's the kind of bad information we have worked many years to overcome and was the reason the trusted cultivator tag was developed in the first place.
RR


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14054780 - 03/02/11 01:53 PM (12 years, 10 months ago)

Quote:

RogerRabbit said:


However, it was the original post with bad information stated as absolute fact that got sand in my panties earlier.
RR




:lol:


--------------------
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Re: Paul Stamets, Endospores, and You [Re: i GrOw StUFF]
    #14055081 - 03/02/11 02:46 PM (12 years, 10 months ago)

Guys i have the answer. You should shoot you're jars into outer space. The radiation will surely kill them all, except Janibacter Hoylei and 2 or 3 others.
But be sure to wear gloves if you want to catch them upon re entry. They can get quite hot.
See thats 2 ways of sterilizing in one package, im a friggin genius.

where's my trusted cultivator tag?


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14055114 - 03/02/11 02:50 PM (12 years, 10 months ago)

Quote:

RogerRabbit said:
Bear in mind, the 121C for 20 minutes figure often sited in the literature is for organisms subjected directly to the steam.  A grain jar or bag isn't the same thing, and the heat doesn't travel through the loosely packed contents of a grain jar the way it would if it was a scalpel or inoculating loop sitting on a tray in the autoclave.  The center of a grain jar or spawn bag is heated by conduction, and this process takes time.




Yes, and that's why we sterilize our jars for 90 minutes and not for 20 minutes. Anyway thanks for your answer RR. I am not trying to be "right": I like to debate with people who know more than I do. :hatsoff:


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14055398 - 03/02/11 03:48 PM (12 years, 10 months ago)

Quote:

RogerRabbit said:
Doc,  you can't just make shit up.

take a biology course at a local community college.

http://www.textbookofbacteriology.net/Bacillus_2.html
http://www.bioinf.uni-leipzig.de/~ilozada/papers/paper_Bacillus/Taxonomy_Genus_Bacillus.pdf




I've taken biology and microbiology, and audited micro a few times too.
http://en.wikipedia.org/wiki/Endospore

Quote:

In contrast to eukaryotic spores, which are produced by many eukaryotes for reproductive purposes, bacteria will produce a single endospore internally.




There's the one-to-one correspondence. I'm not going to go through the rest of your bombast line by line.
I posted what I posted after careful consideration. I hope people will examine the question and decide for themselves-
not take your word nor mine as proof of anything.


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Re: Paul Stamets, Endospores, and You [Re: Doc_T]
    #14055454 - 03/02/11 04:00 PM (12 years, 10 months ago)

Quote:

Doc_T said:

I posted what I posted after careful consideration.




But have you experimented with it, maybe RR believes what he does cause he has hands-on experimented with it enough to know.


Edited by SmokedShroom (03/02/11 04:01 PM)


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Re: Paul Stamets, Endospores, and You [Re: SmokedShroom]
    #14055484 - 03/02/11 04:06 PM (12 years, 10 months ago)

RR claims to have seen endospores reproducing directly.
I wasn't there and I don't know what he saw- but I can say with certainty what he did not see.

I've taken biology and I've taken micro. This is my understanding of endospores.
If you understand it differently, that's great. Rock on.

I'm moving on, y'all have fun with the thread. I hope everybody has learned something.
I've learned how bitter and personal RR can be. "You'll never be a TC! :oldman: ". Like I care- I've never been one so far. :shrug:


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Re: Paul Stamets, Endospores, and You [Re: Doc_T]
    #14055533 - 03/02/11 04:19 PM (12 years, 10 months ago)

yeah my penis is thissssssssssssssssssssssssssssssss big


nice hypothesis Doc_T


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14055950 - 03/02/11 05:31 PM (12 years, 10 months ago)

Quote:

RogerRabbit said:
Doc, you can't just make shit up.

LOL!


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Re: Paul Stamets, Endospores, and You [Re: Doc_T]
    #14056549 - 03/02/11 07:30 PM (12 years, 10 months ago)

Quote:

Doc_T said:
I've learned how bitter and personal RR can be. "You'll never be a TC! :oldman: ". Like I care- I've never been one so far. :shrug:




Not bitter, but when you say Paul or I don't know what we're talking about, and even call Paul down in the thread title, it can't go without being contested.  I'm also pretty sure I've not said you'll never be a TC.

http://www.textbookofbacteriology.net/Bacillus_2.html
Quote:

Kenneth Todar, PhD said:Although many species contain sporangia and free spores within 24 hours, some cultures must be incubated 5-7 days before mature sporangia, and the size and shape of the endospore contained therein, can be observed.




http://en.wikipedia.org/wiki/Endospore ; (Doc's link)
Quote:

When a bacterium detects environmental conditions are becoming unfavourable it may start the process of endosporulation, which takes about eight hours. The DNA is replicated and a membrane wall known as a spore septum begins to form between it and the rest of the cell. The plasma membrane of the cell surrounds this wall and pinches off to leave a double membrane around the DNA, and the developing structure is now known as a forespore. Calcium dipicolinate is incorporated into the forespore during this time. Next the peptidoglycan cortex forms between the two layers and the bacterium adds a spore coat to the outside of the forespore. Sporulation is now complete, and the mature endospore will be released when the surrounding vegetative cell is degraded.




Quote:

Doc_T said:
A bacterial endospore is not a spore or seed, it's an encrusted form of a bacterium. . . .The endospore forms when the live bacterium encounters dry conditions . . .A bacterium becomes an endospore, it does not produce them.




We all make mistakes, but we're here to learn.  It's not an attack on you, rather on the incorrect information posted as fact.  It would be less vitriolic to ask about that which you don't understand, rather than to attack Paul and I for what we've personally observed. 

It's hard to see inside an endospore with a light microscope, but the endospore itself can be easily seen and observed as it forms. They are produced by the cell, not a change of state which will simply revert back to 'the form it had before, a live active bacterium and gets on with business.'. If I had more time, I'd set up my video camera equipped microscope and film it.  Perhaps in the future. . .

Hopefully this clears up any misunderstanding.
RR


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Re: Paul Stamets, Endospores, and You [Re: RogerRabbit]
    #14068107 - 03/04/11 08:12 PM (12 years, 10 months ago)

Quote:

When a bacterium detects environmental conditions are becoming unfavourable it may start the process of endosporulation, which takes about eight hours. The DNA is replicated and a membrane wall known as a spore septum begins to form between it and the rest of the cell. The plasma membrane of the cell surrounds this wall and pinches off to leave a double membrane around the DNA, and the developing structure is now known as a forespore. Calcium dipicolinate is incorporated into the forespore during this time. Next the peptidoglycan cortex forms between the two layers and the bacterium adds a spore coat to the outside of the forespore. Sporulation is now complete, and the mature endospore will be released when the surrounding vegetative cell is degraded.




I had no idea what this means so i found a video:


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