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Offlinedimitri8
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Antibiotic resistance???
    #2830198 - 06/26/04 11:19 AM (13 years, 4 months ago)

Hey.. has anyone ever thought about mutagenizing a fungal strain and testing it for newly acquired antibiotic resistances? I think (being a geneticist) that schouldn`t be too difficult and furthermore it would be amazing protection aganist contams!

anyone got any experience on the field?


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Offlinecharvo
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Re: Antibiotic resistance??? [Re: dimitri8]
    #2830857 - 06/26/04 02:52 PM (13 years, 4 months ago)

whats a geneticist


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Offlinesublimesubmind
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Re: Antibiotic resistance??? [Re: dimitri8]
    #2831016 - 06/26/04 03:48 PM (13 years, 4 months ago)

i thought mycelium was naturally protected against contams, problem is it needs to establish itself before mold or whatever takes up all of the food


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OfflineaNewPerception
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Re: Antibiotic resistance??? [Re: sublimesubmind]
    #2831108 - 06/26/04 04:35 PM (13 years, 4 months ago)

advanced forum??


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Offlinedimitri8
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no... [Re: dimitri8]
    #2839375 - 06/29/04 12:26 PM (13 years, 4 months ago)

No, the idea would be to make the cubensis strain antibiotic resistant and then mixing an antibiotic into the Rice cake... the fungus grows and all contams are killed by the antibiotic....

X-Ray mutagenesis could possibly do that.. anyone got any experience with it?


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InvisibleYidakiMan
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Re: no... [Re: dimitri8]
    #2839687 - 06/29/04 02:01 PM (13 years, 4 months ago)

I've only ever used antibiotic to combat bacteria never a fungi imperfecti contaminant.


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OfflineWebLiZaRD
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Re: no... [Re: dimitri8]
    #2840015 - 06/29/04 03:38 PM (13 years, 4 months ago)

Just don't forget that using antibiotics are really dangerous. The world tries to end the usage of antibiotics. It just make the bacterias or contams stronger. Maybe it works for sometime but mother nature will find another way to contam our cakes. It is the way of nature, you can't save every living thing from death. If you make 100 cakes mother nature will kill 5 or 10 of them and it is not a loss. It is just how nature works.

And using chemical substances for growing something is not a good idea in my opinion. Why don't you decide 100% natural shrooms? Maybe you will get more shrooms with chemicals but it will cause illness in future. When i go to bazaar i prefer chemical-free grown fruits. They are smaller but natural.

I think you should think natural ways to war with contams.
Antibiotics are the worst idea to fight with bacterias because of its usage we have stronger bacterias around.
X-ray usage is another problem in my opinion. It can cause mutants and weird shrooms.

P.S: Just for information i am a veterinary medicine student and we talk a lot about using or not using chemicals. Because of this i can tell you to not to use any chemicals or drugs for growing anything.

Don't forget that there is always nature's way to war with contams. Or better not to war with them. At least they are another living organisms and they need to live like us or shrooms or fruits. If they caught your cakes just give them a feast :=) they will be happy :=)


--------------------
"Come, come again, whoever you are, come!
Heathen, fire worshipper or idolatrous, come!
Come even if you broke your penitence a hundred times,
Ours is the portal of hope, come as you are."

Mevlana Celaleddin Rumi - Alternative spellings: Mawlana, or Mowlana, and Jalal Al-Din, or Jalaluddin
The great Anatolian philosopher, poet and the father of the Mevlevi sect.


Edited by WebLiZaRD (06/29/04 03:40 PM)


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Offlinedimitri8
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Well... [Re: WebLiZaRD]
    #2840674 - 06/29/04 06:44 PM (13 years, 4 months ago)

then you shurely know that antibiotics are in fact being produced (initially) by a fungus in the first place.. so it is natural...

but anyway... Molecular biologists (and genetecists) such as myself use antibiotic resistances every day in the labs. using them to select for the wanted strain (as done in the lab) doesnt make the shrooms "chemical or out of plastic!"

I think I`ll try to produce a resistant strain.... anyone know a readily available antibiotic out there? most I think need a perscription from a doctor....


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OfflineWebLiZaRD
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Re: Well... [Re: dimitri8]
    #2840733 - 06/29/04 07:00 PM (13 years, 4 months ago)

i know it seems natural. I just want to mean that it is not what the nature did. It is human-made thing.
I just think another way to fight with contams. Maybe there is another fungus or any living organism eats green mold (for example) but never hurts magic shrooms. These kind of usage is better. I will search for this kind of usage. It is like this "When you see rats at home, don't try to poison them. some of them will get poisoned but some of them will survive and births stronger baby rats. Instead of this just feed a cat and it will eat all of rats :smile:"


--------------------
"Come, come again, whoever you are, come!
Heathen, fire worshipper or idolatrous, come!
Come even if you broke your penitence a hundred times,
Ours is the portal of hope, come as you are."

Mevlana Celaleddin Rumi - Alternative spellings: Mawlana, or Mowlana, and Jalal Al-Din, or Jalaluddin
The great Anatolian philosopher, poet and the father of the Mevlevi sect.


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Offlinedimitri8
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Re: Well... [Re: WebLiZaRD]
    #2842610 - 06/30/04 06:30 AM (13 years, 4 months ago)

I know of the accumulation of antibiotic resistances... they are a problem... but humanity will use them untill they are useless and then find an alternative....

and antibiotics are not a man-made thing! ever heard of penicillin? It originates from a natural mold that secretes the antibiotic to kill bacteria/contaminants.... if only the penicillium fungus were to produce psilocybin all this contamination would not be a problem... and thats the idea of this thread.... make cubensis antibiotic resistant and add penicillin as the penicilum mold would...


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OfflineWebLiZaRD
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Re: Well... [Re: dimitri8]
    #2842656 - 06/30/04 07:00 AM (13 years, 4 months ago)

ok i think i got the point. Sorry for my poor english, i hope i didn't make you angry.
I know about penicillin and that is what i exactly want to explain. Such a thing like penicillin would work and it will be really great.
We are on summer holiday and after holiday i will start to look for your aim at university. In my university we have a prof. he is one of the best mycologist in department of plant protection. i will go and talk with him. maybe get some help.

i really like researches like you do. You know how penicillin discovered. Hope you will discover a new method to prevent from contams. My only thought is it would be a natural way. I never try to make your thoughts change. Just want to share thoughts. :=)
I hope i can explain my thoughts truely.

Good luck and hope you will win the battle :smile:


--------------------
"Come, come again, whoever you are, come!
Heathen, fire worshipper or idolatrous, come!
Come even if you broke your penitence a hundred times,
Ours is the portal of hope, come as you are."

Mevlana Celaleddin Rumi - Alternative spellings: Mawlana, or Mowlana, and Jalal Al-Din, or Jalaluddin
The great Anatolian philosopher, poet and the father of the Mevlevi sect.


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Invisiblemicro
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Re: Well... [Re: dimitri8]
    #2844320 - 06/30/04 06:54 PM (13 years, 4 months ago)

Quote:

if only the penicillium fungus were to produce psilocybin all this contamination would not be a problem... and thats the idea of this thread....




Fungi are already resistant to bacteriostatics -- I'm suprised you didn't know this, "being a geneticist" and all.... Myself, I've lost many antibiotic plates to fungi....

The only thing that might help would be fungicide resistance -- I know I posted something in this forum before.... If you are really serious, look up some genes for fungicide resistance in Saccharomyces cerevisiae or Neurospora crassa in PubMed. Random mutations would not work with a lot of fungicides because of their physiology (i.e. inhibition of the electron transport chain) but maybe with some.... You'd have to look it up in a mycology text and decide for yourself.

Some ref's maybe:

http://www.ncbi.nlm.nih.gov/entrez/query...t_uids=12619694
http://www.ncbi.nlm.nih.gov/entrez/query...t_uids=12612806
http://www.ncbi.nlm.nih.gov/entrez/query...t_uids=12450134
http://www.ncbi.nlm.nih.gov/entrez/query...t_uids=12233175
http://www.ncbi.nlm.nih.gov/entrez/query...t_uids=12135574
http://www.ncbi.nlm.nih.gov/entrez/query...t_uids=12061897

etc., etc. These were all from a search for fungicide resistance gene @ Pubmed.org

--
Micro


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Offlinedimitri8
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Re: Well... [Re: micro]
    #2846651 - 07/01/04 11:01 AM (13 years, 4 months ago)

interesting replies! :smile:

well... actually introducing a plasmid with fungicide/antibiotic resistance would be a solution of course... if also somewhat more complex.... I know that many fungi and bacteria are allready resistant to antibiotics.... and antibiotic agar is often contaminated by such resistant fungi or bacs, especially in labs where resistant strains are made and then escape into the surroundings to wait of an unprotected plate...

well.... fungicide would possibly protect against other fungi, but not other bacteria! while antibiotics (as long as no resistances have developed) would combat every kind of contamination, so I thing that would be a more interesting candidate for this... a cocktail of several substances in high concentrations would of course be the best case, as no resistances would develop in contams.... in theory this could go as far as not needing to use sterile techniques or pressure cooking sterilization!

I`ll see what i can achieve... :smile: and then let you know! :smile:


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Invisiblemicro
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Re: Well... [Re: dimitri8]
    #2847291 - 07/01/04 02:16 PM (13 years, 4 months ago)

Quote:

well.... fungicide would possibly protect against other fungi, but not other bacteria! while antibiotics (as long as no resistances have developed) would combat every kind of contamination, so I thing that would be a more interesting candidate for this...




I think you are not understanding me, or the way that antibiotics work. Bacteriostatics (like pen, strep, tetra, etc, etc.) effect bacteria. They don't effect fungi, anyway, because of the way they work -- not because of a specific resistance. You can add a bacteriostat to a petri and grow fungi off of it, already -- you don't need to have a specific resistant strain.

For example penicillin works by inhibiting the formation of bacterial cell walls by interfering with their transpeptidase enzymes that form the cross links between peptidoglycan strands of the cell walls. Since fungi lack these enzymes (in addition to the same type of cell wall -- fungal cell walls are something like 50% chitin) it does not effect them. Other antibiotics are similar -- the bacteriostatics are not antifungals -- the antifungals is a different class of antibiotic, altogether.

Now, antifungals are a different story. Mycelium should not easily grow on a plate with a general type of antifunal -- a specific gene must be either inserted to effect resistance or some other method must effect resistance -- such as selective pressure, but, like I said, it would have to be likely to happen because of a random mutation, so you'd have to look up different fungicides and their methods of action. Two antifungals to which fungi easily develop a resistance are flucytosine and griseofulvin -- take it for what you will.

--
Micro


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(Avatar is Maxxy, a character by Mizzyam, RIP)


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Offlinetubervegan
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Re: Antibiotic resistance??? [Re: dimitri8]
    #2852001 - 07/02/04 05:46 PM (13 years, 4 months ago)

What you're are doing sounds interesting and everything, but it also seems kind of pointless. You could make cakes with a anti-biotic/anti-fungal cocktail + the anti-fungal resistant strain of your mushroom, but you'll notice that is pretty much exactly the same as the peroxide tek. Remember spores cannot produce superoxide disumutase and catalase so peroxise will kill them, by using already spawned mycelium in a peroxide cake you can accomplish the same thing you are trying to do... in theory. I've found this technique doesn't work.

So back to anti-fungal resistance. The only clean way to do this would be with some sort of plasmid vector (I'm not sure how plasmids work with fungus), using x-ray mutagenesis would be difficult to use on mushroom I think because the sexes aren't well defined. If you can separate the positive and negative types then you might be able to go somewhere with it, otherwise even if you happen to get a resistant strain you wouldn't really be able to trace your results, but I mean pragmatically it would work.

That said, good f***ing luck. For one, if you are going to try to make fungicide resistant plasmids or mushrooms, do it for regular buttom mushroom because you'll make a fortune selling the patent, instead of a moderate amount of convienence when growing cubensis. and second I don't see how you are going to beat the fungicides out there nystatin attacks the chitin walls, flucytosine attacks DNA/RNA transcription/translation and griseofulvin inhibits growth (not too sure on the mechanisms) they are all really generic and the only real way I could see this being done is via gene splicing with entirely different species, like I don't even mean mushrooms, and I don't know if thats even possible due to that nature of mushrooms being like pseudo-eukaryotics. SO anyways i'm now just ranting but my advice is either perfect the peroxide tek or prepare to dedicate your lifes work to this.


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Offlinefelixhigh
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Re: Well... [Re: dimitri8]
    #2852407 - 07/02/04 09:02 PM (13 years, 4 months ago)

Quote:

dimitri8 said:
in theory this could go as far as not needing to use sterile techniques or pressure cooking sterilization!





whats the problem with sterilization/pasteurization? it's boring and worksome? yeah, but as a geneticist you should know thats part of it... perhaps you could work up your sterilization technique?
anyways, the concept is interesting. i mean, taking off the 'non sterile substrate' part. as a geneticis you should have already seen that some batches and strains are more or less resistent to contams than others and eventually find one that comes out to 'eat' the contams... i believe one could isolate such sectors (yuck) and then test them for fruiting habilities, reisolate and so on and on... but i wouldn't bet on a raw substrate eating mushroom... anyways watch out, don't go make a glass eating mycelium that will burrow through the earth out of control...


FH :mushroom2:


Edited by felixhigh (07/02/04 09:09 PM)


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Offlinetubervegan
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Re: Well... [Re: felixhigh]
    #2854061 - 07/03/04 03:18 PM (13 years, 4 months ago)

It all just seems pointless to me, if your jars are contaminated you're going to toss them (I'd hope), regardless of whether or not what you are growing can eat the other mold or resist them. You're still going to get a mushroom full of potentially and probably toxic metabolites. You'd be better of trying to find a remedy to cure people when they get toxic shock from eating the mushrooms.


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Invisiblefastfred
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Re: Well... [Re: tubervegan]
    #2904639 - 07/19/04 08:30 AM (13 years, 4 months ago)

> For one, if you are going to try to make fungicide
> resistant plasmids or mushrooms, do it for regular
> buttom mushroom because you'll make a fortune
> selling the patent, instead of a moderate amount
> of convienence when growing cubensis.

Too late, it's already been done. Of course, that would make it real easy for a top-notch geneticizst like yourself.

Here is a link on Oyster resistance...
http://tinyurl.com/4kmf3

It's also been done with A. bisporus


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Offlinedimitri8
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Very very interesting.... [Re: fastfred]
    #2985411 - 08/10/04 12:05 PM (13 years, 3 months ago)

hey... I am surprised about the high standard of all the replies.... thank you all.. it has been very interssting.

To the practicality of it all.... Well.. sure it is possible to culture the shrooms without all this antibiotic stuff, but wouldnt it be nice to just have to mix up your substrate, pour the mycelium culture onto it and see 100% grow without contamination? No syringes, no lighters, no peroxide and 100% success? Of course it is not a necessity, but in my opinion it would make it all a little easier... hence my interest in improving things.

Of course there are some antibiotics that loose their function bec ause the protein it atacks has randomly mutated enough to become resistant.. and others that have such a complex mode of action that random mutagenesis would only very unlikely produce a resistant strain.... and that of course has to be taken into account.
There are loads of resistance genes available in molecular biology laboratories and a plasmid vector carrying this gene could probably easily be inserted into the fungus. the more complex the resistance is, the more difficult it is for bacs to develop resistances against it.

Ok.. I was not aware, that antibiotics only harm bacteria, and I was under the impression that they for example also harm plant cells, as they are (to my knowledge) also sometimes used in biolistics. But it doesnt matter. all i have to do is find an anti fungal agent, an antibiotic and the corresponding resistance genes, put them in a vector and transform the strain. Select for transformants by the action of the antibiotic and anti fungal agent and voila. a resistant strain.


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OfflineKonkeyDong
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Re: Very very interesting.... [Re: dimitri8]
    #2985603 - 08/10/04 01:01 PM (13 years, 3 months ago)

Dr. Rush Wayne wrote a couple books about using peroxide as an anti-contam. You have to use live myc. cultures and it is pretty easy to do. But, I just use a pc and do it the old school way.


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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