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InvisibleHippie3
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: micro]
    #1796147 - 08/09/03 07:57 PM (18 years, 5 months ago)

clumped is just another word for concentrated,
thus the principle remains the same.
a concentration [clump] of spores can locally over-whelm the peroxide and germinate.
like i said way back a few posts ago..


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Invisibleafoaf
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: Hippie3]
    #1796168 - 08/09/03 08:05 PM (18 years, 5 months ago)

a friend recently drew up a cc or 2 into a spore
syringe, mistaking it for a culture syringe.

(lesson: no smokey while workey)

afoaf told him, well, good luck, I think you've
killed her.

he came back a few weeks later and said that
he had knocked up some brf jars just for shits
and giggles and that they had germinated and
were showing PHENOMENAL growth.

true story.


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Invisiblemicro
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: afoaf]
    #1799327 - 08/10/03 09:18 PM (18 years, 5 months ago)

Yaeh, Hippie -- thanks for your vague post. You'd get 1/2 cerdit if I were grading you on a test....

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Micro


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Invisiblemah2b1
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: micro]
    #1800002 - 08/11/03 01:08 AM (18 years, 5 months ago)

Hippie3 has a good analogy, and this statement in Dr Rush?s e-mail to Micro backs it:

?All organisms produce the enzymes, but the isolated ones are easily overwhelmed by peroxide, whereas the multicellular ones can defend themselves against it.?

Appaerently there are peroxidase enzymes inside the spores, which will decompose the peroxide, but the spore is destroyed in the process. The spores are not fully alive like the mycelium, which can produce its own peroxidase enzymes as needed.

The peroxidase enzyme can only decompose a certain amount of peroxide before it gets burned out and stops working. Therefore, using a large clump of spores will eventually create an area of non-peroxidated agar media for the unaffected spores (those protected inside spore clumps) to germinate.

Assume each spore had one peroxidase, and that peroxidase could catalyze one molecule of peroxide. You take a spore clump containing 150 spores; 100 spores exposed on the outside of the clump and 50 contained within. You place the spore clump on peroxidated agar. The spore clump occupies a surface area containing 100 molecules of peroxide. The 100 spores on the outside of the clump are destroyed while decomposing the 100 molecules of peroxide present in that area. This leaves that area of agar media without any peroxide, yet there are still 50 spores left that can germinate and mate with each other. Therefore, the spore clump has OVERWHELMED the peroxide in the agar.

The peroxidase enzyme and peroxide are similar to Hippie3?s matter and anti-matter. They both will eventually cancel each other out.


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Invisiblemah2b1
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: mah2b1]
    #1800019 - 08/11/03 01:10 AM (18 years, 5 months ago)

As for AFOAF, the spores syringes made with peroxide probably underwent the same process, leaving behind only a few viable spores that were protected in clumps. Those clumps went on to germinate.

There could be many explanations for the aggressiveness of the mycelium produced. One may be that there were less sub-strains produced (from the viable spores mating) that would compete amongst themselves for available substrate. Or, the use of the peroxide (antibiotic chemical) weeded out the bad spores leaving behind the stronger more viable spores for mating after the peroxide was decomposed. These are only two reasons for your friend?s success, after all your friend botched up the syringes to begin with, but it is an interesting bit of information.

What was the concentration/formula of the peroxide solution used to make these accidental spore syringes (may come in useful when working with contaminated prints)?


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InvisibleHippie3
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: micro]
    #1800937 - 08/11/03 09:36 AM (18 years, 5 months ago)

suit yerself,
you can count on that being the very last time i waste my time trying to educate you.
i teach you something you didn't know,
Quote:

Neat! Thanks, I didn't know that.



and you then repay me with this shit ?
Quote:

Yaeh, Hippie -- thanks for your vague post. You'd get 1/2 cerdit if I were grading you on a test....






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Edited by Hippie3 (08/11/03 03:01 PM)


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Invisiblemycofile
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: Hippie3]
    #1801473 - 08/11/03 01:51 PM (18 years, 5 months ago)

This has gotten silly. The answer to the original question/post, as hippie just pointed out, is that spores can germinate in the presence of peroxide, and we have some practical guidlines for how to make that work.

I really didn't mean to sow the seeds of dissent against hippie, I just like to be pretty careful in describing how I think something works vs the way it's been proven to work. I think hip's original anologies of overwhelming clumps and matter/anti-matter are good anologies for a general understanding of the practical applications. But I've not seen scientific proof that it's exactly right. It very well could be a matter of strains whose spores are more peroxide resistant being selected, like with bacteria and antibiotics. I don't really think so, I think hippie and Wayne are on the right path, but I just haven't seen proof of it, and I only leave the possibility of Wayne being wrong because he stated in his first book that this was impossible.

And yes, this is old news. The method was first brought to my attention back when drooldonkey first opened, so that was years ago.

And yes, it's not necessary, mycophreak talks about using slants, and simply flaming the lip before opening. Not to mention, I know plenty of people who have decent success doing open air agar. Especially when you use peroxidated agar for the 2nd and subsequent plates, the first one doesn't NEED to be peroxidated.

So, unless somebody does have a truly scientific breakdown of the processes involve, then there is no need arguing. It can be done. We know how to make it more likely to be successful (high spore concentration). Why argue?

Oh and to
Quote:

If I have only one print of a species, would you recommend heavy streaking on peroxidated agar or light to normal streaking on regular agar? First time agar user here.



Since a print has plenty of spores to streak hundreds of plates, peroxidated or not, I wouldn't recomend one over the other. It's just a matter of your personal situation. I mean if you've got a flow hood, you wouldn't use the peroxide. Either way you should be succesful if you follow the proper guidlines and principles.


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Invisibleafoaf
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: mah2b1]
    #1810838 - 08/13/03 08:40 PM (18 years, 5 months ago)

it wasn't my intention to imply that growth
was phenomenal because of the peroxide.
moreso, it was phenomenal in spite of the
peroxide in the syringe.

it was a 10cc syringe, he prolly drew up an
additional 2 cc's of 7% peroxide.

it was definitely a dark syringe and the strain
tends to be quite voracious during colonization.

maybe just enough surivived to keep hope alive.


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Invisiblemycofile
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: afoaf]
    #1813012 - 08/14/03 11:58 AM (18 years, 5 months ago)

Yeah, good clarification, in spite of rather than because, I was a little confused about exactly what you meant too.


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PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.


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Invisiblemah2b1
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: afoaf]
    #1814910 - 08/14/03 09:17 PM (18 years, 5 months ago)

Thanks for the feedback AFOAF.

I have some thick prints that I know are contaminated. I am hoping that this spore clump protection idea will work with mild peroxide treatment.

I am figuring that the contams are only on the outsides of the spore clumps, and that if I hydrate them with water and then treat them with peroxide that it will kill off most of the contams. This should give me a better chance to obtain some clean mycelium for a tissue transfer.


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Invisiblemycofile
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: mah2b1]
    #1816790 - 08/15/03 01:49 PM (18 years, 5 months ago)

Yeah, but you could just germinate them on agar, then isolate the mycelium for the transfer. If it's just a matter of not wanting to mess with agar, BRF paste is a good alternative. The flat surface of these substrates just makes the isolation and transfer a lot easier.


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"From a certain point of view"
-Jedi Master Obi Wan Kenobi

PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.


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Offlinemickywilliams2
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: mycofile]
    #2129389 - 11/22/03 06:57 PM (18 years, 2 months ago)

The peroxidase enzymes came from the gills of the mushroom and are just stuck to the spores.





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OfflineAnnoA
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: mickywilliams2]
    #2129867 - 11/23/03 02:15 AM (18 years, 2 months ago)

Fact or guess?


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Offlinemickywilliams2
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: Anno]
    #2130320 - 11/23/03 12:28 PM (18 years, 2 months ago)

guess



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Invisiblemicro
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: mickywilliams2]
    #2130480 - 11/23/03 02:25 PM (18 years, 2 months ago)

Spores produce enzymes -- they clump together and can produce more enzymes for a given volume than something like bacteria (or isolated spores) could because of the fact that they clump together. When it's in a liquid, for example, it obviously can't exhaust all the peroxide in the area around it -- it just neutralizes it.

This confuzed me for a while.... It's the clumping of spores or the organization of the hyphae that enables it to exhaust the peroxide -- it doesn't work for bacteria, because of this.

Oh, and I hate to beat a dead horse, but I forgot about this post. I didn't mean to start an argument, but Hippie -- what you said:

Quote:

and my matter/anti-matter analogy applied only to spores and peroxide, so the enzyme factor wasn't the point since that only applied to living tissue, not spores, which is what afterall, i was talking about.




is implicitly wrong -- that's why I meant your post was vague. The spores do produce peroxidases, and they clump, so they're able to knock out the peroxides in a given area because they produce more enzymes per given area than a bacteria, for example, since they do not clump.

Oh, yeah, and:

Quote:

"clumped is just another word for concentrated,
thus the principle remains the same."




No, it is not. Spores clump -- bacteria does not.

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OfflineElemicin
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: micro]
    #2131419 - 11/24/03 12:28 AM (18 years, 2 months ago)

depends on the concentration :smile:.....

high percentages of H2O2 are known to cause organic matter to explode....(we're talking 60+%)






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Invisiblemicro
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: Elemicin]
    #2134321 - 11/25/03 08:40 AM (18 years, 2 months ago)

I would assume we're talking about the concentration used in agar, since that's what this thread is about....

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Offlinedebianlinux
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Re: Does H2O2 KILL spores or just inhibit germination? [Re: micro]
    #2137195 - 11/26/03 11:24 AM (18 years, 2 months ago)

Quote:

micro said:
Quote:

hippi3 said:"clumped is just another word for concentrated,
thus the principle remains the same."




No, it is not. Spores clump -- bacteria does not.

--
Micro




While I have no wish to enter this argument I do have to agree with micro on that one. Bacteria may be concentrated but are in no way "connected" to one another except during mitosis(?). The spores, OTOH, actually web/knit themselves together with mycelium, as is their nature. That is all.


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