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InvisibleOlgualion
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wild P. mexicana - strange growth on agar?
    #1314333 - 02/17/03 12:45 PM (21 years, 5 months ago)

Dude obtained a wild P. mexicana print a while back. He innoculated agar and transferred the best sections. after 3 transfers the growth was all white. He then transferred wedges to some rgs. A few weeks later he looked at the jars and they had all begun to get this black mold growing on them, then looking at the dishes he noticed that what first appeared to be the beginnings of sclerotia now had the same black growth on them. We didn't now what to think, so he made a few more transfers. Finally he had a plate grow out which was pure white and appeared to be healthy, he transferred this to more peroxidated agar.

Here is the strange part. I'm sure you know how peroxidated agar gets a cloudy appearance around the outside of the perimeter of the growing mycelium. Well, this plate has had that cloudy appearance grow all of the way to the edge of the plate now (bacteria?); The mycelium wedge is beginning to grow over top of it!!

It makes me think that bacteria was hitching a ride with the wedge, it first colonized the peroxidated agar, then the mycelium began eating the bacteria?? I really don't understand it. Has anyone seen anything like this, I am totally confused. If you can't understand my rambling, please ask me to clarify. I feel like we are getting very close with this wild mexicana, but it is just out of reach.

Finaly, what he did was transfer a few pieces to new plates below the agar per Dr. Wayne Rush.

What do you all think? Have you seen growth like this before?


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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1315278 - 02/17/03 06:09 PM (21 years, 5 months ago)

The mexicana I grew out on plates started as a very fine layer of mycelium and then bulked up to more cottony growth. It is not quite what you described though.

Good luck!

Joshua


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InvisibleChampion des Champignons
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1315324 - 02/17/03 06:34 PM (21 years, 5 months ago)

Quote:

it first colonized the peroxidated agar, then the mycelium began eating the bacteria??



more likely the outer edge of the mycelium broke down the h2o2, allowing the bacteria to get a foothold.
unless it's a yeast?
Not sure if yeasts have the ability to break down h2o2, but I suspect so...


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InvisibleOlgualion
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Re: wild P. mexicana - strange growth on agar? [Re: Champion des Champignons]
    #1315991 - 02/18/03 04:57 AM (21 years, 5 months ago)

Hmmm... maybe yeast, I've never seen bacteria like this. But then again maybe it is something indiginous (sp?) to Mexico.

Wish I had a camera. Or even my copy of 'TMC' - I think I've bought the book 3 times and ended up losing it one way or another each time..

The bacteria?, yeast? seems to have no problem decomposing the peroxide. The mycelium has just begun growing over top of it. The contam is very "slimy like". In fact the plate was kept upside down and whatever it is has actually dripped down to the top of the plate (that sounds funny). I guess I'm really just curious about what it is for sure. I really wish I had a camera... Hopefully soon...


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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1318607 - 02/19/03 05:50 AM (21 years, 5 months ago)

"I'm sure you know how peroxidated agar gets a cloudy appearance around the outside of the perimeter of the growing mycelium."

I do not know this. What kind of agar are you using? The clouding of your agar is likely due to the digestion of sugars and unrelated to the peroxide.

Joshua


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InvisibleOlgualion
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Re: wild P. mexicana - strange growth on agar? [Re: Joshua]
    #1320280 - 02/19/03 04:47 PM (21 years, 5 months ago)

hmmm...  I could just be seeing things that aren't really there. :wink:  It has happened before, and im sure will happen again...  But really though, peroxidated agar seems to have a cloudy appearance just before that area begins colonizing *especially on the bottom of the plate*.  I dunno...

Anyway, this nasty stuff doesn't want to go away.  Even more strangely, the mycelium seems to eat it up.  We'll see how this most recent transfer will go shortly.  Do you think adding gentamycin to the agar will help?  Will it keep away a yeast if that is what this is? 


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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1320328 - 02/19/03 05:09 PM (21 years, 5 months ago)

I use DFA which is not transparent. I have worked with bacterial cultures grown on various sugar agars. The plates will cloud or clear in the presence of a bacteria that can metabolise the given sugar. I have some better agar I will pour my next time around to see if I can duplicate what you are seeing. What type of agar do you use?

Yeast are in the fungi family. I think most if not all fungi can break down peroxide. Most bacteria can not. Adding gentamycin is primarily to protect from bacteria.

I am curious as to what you are seeing. Do you have a way to get a pic?

Joshua


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InvisibleOlgualion
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Re: wild P. mexicana - strange growth on agar? [Re: Joshua]
    #1320462 - 02/19/03 06:03 PM (21 years, 5 months ago)

This is the best dude could do with a crappy scanner:


The blobs you see are from the contam dripping while the petri was upside down. It isn't very easy to make out because there is some condensation as well. Hopefully you get the idea enough to make a diagnosis...

If you look really hard, you can see where the agar has actually split, the contam somehow caused this. The agar is homemade peroxidated PDA.


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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1320488 - 02/19/03 06:27 PM (21 years, 5 months ago)

My first thought was that the starch from your mix is seeping out of the agar. Don't take it too seriously though. That is quite a bit of mass for a contam. I have never seen anything like that before. Is there anyway for you to narrow the contam to something other than the culture? Is the contam on all of the plates of this batch? Is the agar you used on this transfer any different than the previous transfers? Does the plate smell funny?

Joshua


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InvisibleOlgualion
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Re: wild P. mexicana - strange growth on agar? [Re: Joshua]
    #1320518 - 02/19/03 06:46 PM (21 years, 5 months ago)

Same exact batch of PDA has B+ and a P. tamp (iso), growing nicely; also unperoxidated from the same batch is P. tamp from spores, and they are just beginning to germinate.  Only this wild P. mex has this nasty snotty stuff.  Some chunks of mycelium were taken from this plate and bottom innoculated into plates with two other batches of agar (PDA and MYA +a bit of poo).  The PDA ones already are showing this contam, while the MEA hasn't done anything yet.  So it is definitely not the agar, and I haven't dared sniff it, and probably won't.. :wink:

I would certainly say it is not a mold!  right?  The original print was terribly contaminated with numerous molds I've never encountered before.  The funny thing is that the plate this one was transferred from appeared clean (and still does), I thought it was clean until this stuff showed up.

--edit--
Now that i think of it, this batch had nutritional yeast added(other than that, it is the same recipe), but like I said the other species are doing fine on it...


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Edited by Olgualion (02/19/03 06:50 PM)

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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1320578 - 02/19/03 07:39 PM (21 years, 5 months ago)

My only other thought is that it is a slime mold. It is indeed perplexing.

Joshua


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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1320595 - 02/19/03 07:52 PM (21 years, 5 months ago)





Joshua


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InvisibleOlgualion
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Re: wild P. mexicana - strange growth on agar? [Re: Joshua]
    #1320943 - 02/20/03 02:41 AM (21 years, 5 months ago)

So mold can actually grow like that, hmm.. I really need to get some mycology/ bilogy books so I can bring this hobby to a new level. Any you recommend?


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Edited by Olgualion (02/20/03 07:26 AM)

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InvisibleJoshua
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Re: wild P. mexicana - strange growth on agar? [Re: Olgualion]
    #1321443 - 02/20/03 06:34 AM (21 years, 5 months ago)

I have learned what I know through my studies of biology. I have read very few mycology books.

Joshua


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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: Joshua]
    #1371727 - 03/13/03 05:46 AM (21 years, 4 months ago)

Dude went and trashed the other cultures.  I am actually thinking the growth was not a contam, and like joshua said, something to do with the agar.  For some reason not all of the plates were affected though.?. 

Anyway, dude went back to his old multispore plate and squirted 2 cc's of straight 3% H2O2 on top.  Then he cut out a few sections and transferred three pieces to two different plates.  Here is the resultant growth from one of them:


The other plate has some green mold starting, but he is out of plates, and will not be getting more for a few weeks :frown:, so he will put it into the fridge to slow growth and transfer when he can get more.  I am soo stoked that he finally has gotten some good growth from this wild print, now will begin the process of taking more isolates and attempting to fruit them.  He also saved the original culture after dumping out the H2O2, and will take a bunch more sections from it once he has more plates. 


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InvisibleJoshua
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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1372076 - 03/13/03 07:52 AM (21 years, 4 months ago)

I had a dream last night or the night before about slime mold. I tried to take a picture for you.

Joshua


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Offlinejarhead
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Re: wild P. mexicana - finally some nice growth! :) [Re: Joshua]
    #1384141 - 03/17/03 11:25 AM (21 years, 4 months ago)

i start keeping my agarcultures upside down for no water dripping on the mycelium and it seems to work fine , i use babyjars covered with alufoil and used it for a tissuetransfer

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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: Joshua]
    #1385225 - 03/17/03 04:18 PM (21 years, 4 months ago)

:smile:

I actually came across a web site just the other day (wish I could remember it now) that had some pics of this blood red colored slime mold.  It looked like a bunch of droplet sized blood red eyes with blood colored stalks attached.  They were all in a line.  It is tough to describe, but nevertheless looked really neat!

Jarhead, that is a good idea.  I used to use half pint mason jars when I didn't have petris, and they worked well.  The one problem I ran into though is that the bottom is convex and I didn't pour enough agar to cover the center. 


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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1385773 - 03/17/03 06:46 PM (21 years, 4 months ago)

Are you sure that is a Psilocybe mexicana? It looks more like a cubensis mycellium to me. All my mexicanas have multicolored zonate myc.

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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: zeronio]
    #1386603 - 03/18/03 02:59 AM (21 years, 4 months ago)

Honestly, there is a possibility that it is not.  My bud can't remember innoc'ing any plates with cubensis in a long time.  He found the donor plate and it was labeled P. mex and dated (along with a ? mark) shortly after he received a few wild P. mex prints.  There is a possibility that it is a cubie, but I really hope not.  Grain will be innoced with wedges, and it will be grown out soon.  Dude would have to go back to the original contam'd print if it is not mex, but that is okay because he is getting a case of 500 petris soon! :smile:

Now that I look back, this would be the 4th or 5th transfer from the original multispore innoc.  He has no other wild P mex plates, and I can't see him losing what he had worked on for so long.  There was a time he was running very low on plates, but I still can't fathom it.

We'll see, and I'll keep you updated. 


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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: zeronio]
    #1386607 - 03/18/03 03:00 AM (21 years, 4 months ago)

Out of curiosity, where did your P. mex come from?


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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1386654 - 03/18/03 03:25 AM (21 years, 4 months ago)

Some came from MushMush. The mexicana, tampanensis & atlantis have a beautiful and very similar looking mycelium. They're pictured in "The mushroom cultivator".

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Re: wild P. mexicana - finally some nice growth! :) [Re: zeronio]
    #1386785 - 03/18/03 04:20 AM (21 years, 4 months ago)

My buddy has experience with tampanensis, and I have posted a few pics.  I love the orangish color that it has, and IMO it is a very beautiful mycelium (my favorite). 

My bud has a few bags of r.g.s. colonising with mex A strain, and all I have noticed is white.  Maybe the other colors come more with age?  This is not the case IME with tampanensis, which upon spore germination quickly shows its colors.

I haven't ever seen any P. atlantis mycelium, but would love to.  Do you have a print? :wink:  Could you post a pic of some your mexicana mycelium on agar? 


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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1386829 - 03/18/03 04:37 AM (21 years, 4 months ago)

P. mexicana on PDYA:


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Re: wild P. mexicana - finally some nice growth! :) [Re: Anno]
    #1387256 - 03/18/03 07:11 AM (21 years, 4 months ago)

P. mexicana on peroxidated DFA. Note sclerotia formation and lack of multi-colored zonation.



The zonate comment was intended for zeronio

Joshua


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Edited by Joshua (03/18/03 07:16 AM)

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Re: wild P. mexicana - finally some nice growth! :) [Re: Anno]
    #1388880 - 03/18/03 04:09 PM (21 years, 4 months ago)

thanks for the pics guys!

Joshua, your pic makes me think of an idea I had in the past when playing with the tamps. Notice how the sclerotia seem to form more readily when they reach a non nutritious barrier ie. the plastic separations of the dish, or the sides of a glass jar... Well this made me think that maybe the addition of pieces of rock, or marbles, or some other such non nutritious additive would help with formation of stones say in the center of a bag of r.g.s. What do you guys think?

What made me think of this was when dude separated through his old tray of tamp substrate he noticed that almost all of the stones that were inside the mycelial block were formed on and sometimes even in sunflower seed shells (a non-nutritious or at least minimally nutritious "barrier").


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Edited by Olgualion (03/18/03 04:13 PM)

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Re: wild P. mexicana - finally some nice growth! :) [Re: Joshua]
    #1389526 - 03/18/03 09:07 PM (21 years, 4 months ago)

Yes, the zones are not always very obvious. I think that they occur because of different speed of growth of mycellium. At least my cultures have periods of faster and slower growth.
I'm going to try to make a time lapse video next time.

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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1390224 - 03/19/03 04:26 AM (21 years, 4 months ago)

I was actually thinking the exact same thing, seeing as stones are usually found in a casing layer (non-nutritious).. it leads me to believe if you added a large ammount of non-nutritious substrate you might have higher stone formation levels. Marbles would probably work well, or if you could add something organic and absorbant that has no nutrition (something similar to coir maybe) to your substrate, that might work too?


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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1390225 - 03/19/03 04:26 AM (21 years, 4 months ago)

I was actually thinking the exact same thing, seeing as stones are usually found in a casing layer (non-nutritious).. it leads me to believe if you added a large ammount of non-nutritious substrate you might have higher stone formation levels. Marbles would probably work well, or if you could add something organic and absorbant that has no nutrition (something similar to coir maybe) to your substrate, that might work too?


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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: Raadt]
    #1390481 - 03/19/03 05:55 AM (21 years, 4 months ago)

The next time dude cooks up some bags, I will ask him to try this. I feel that a bag of r.g.s. has a max potential yield depending upon the substrate (obviously). If the addition of non-nutritious 'barriers' give more points for sclerotia to begin, I can see the turn around time to reach this potential being greatly reduced.

I am calling them barriers, because IMO what the mycelia is doing is colonising substrate, then once the growing mycelia reaches these obstacles, it thinks 'hey, since I can't grow out anymore, I will put my energy into producing stones'


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Edited by Olgualion (03/20/03 02:44 AM)

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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1393897 - 03/20/03 02:44 AM (21 years, 4 months ago)

Thoughts anyone?


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Offlinekayaman9
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Re: wild P. mexicana - finally some nice growth! :) [Re: Olgualion]
    #1396724 - 03/21/03 12:48 AM (21 years, 4 months ago)

Just wondering if that is from the wild print I sent you? .

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InvisibleOlgualion
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Re: wild P. mexicana - finally some nice growth! :) [Re: kayaman9]
    #1396900 - 03/21/03 02:25 AM (21 years, 4 months ago)

No, this came from a nice trade with someone south of the border.

I'm going to PM you.


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Re: wild P. mexicana - finally some nice growth! :) [Re: zeronio]
    #1403555 - 03/23/03 01:39 PM (21 years, 4 months ago)

I was looking through some of my old posts, and came across a post with this pic


I now remember that this plate grew for a while then all of a sudden started getting black contamination that would quickly envelope the mycelium upon sporulation. The pictured plate is (i think) 2 transfers ago. It was an attempt to make an isolate from what dude and I thought to be a stone. If I remember correctly it came from a jar which was innoculated with a wedge. I had thought the culture was clean at that point, but subsequently found out that there was a hiding contam. The stone/ growth was halved, washed in H2O and placed onto agar in two different plates. The one plate contam'ed and this is the other. The only way this could be anything other than P. mexicana is if it is contaminated with P. cub. Which I suppose is possible. But I would think remotely.

Anyway, dude innoced a few jars with wedges, and they are nearing 50% colonisation after 6 days. He will probably shake for the second and final time tomorrow. I am getting all anxious to see how they will fruit.

Watch them be cubies now... lol


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