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Invisiblemicro
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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22341024 - 10/06/15 01:03 PM (8 years, 7 months ago)

*nods*

Since they are supposedly able to utilize it as a sole carbon source that would be a good way to isolate it if needed, of course.

I'm sure you knew that but figured I'd point it out in case it helps anyone.


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Re: Pestalotiopsis microspora [Re: micro]
    #22341894 - 10/06/15 05:39 PM (8 years, 7 months ago)

Awesome Cloudy! Can't wait to see results. I haven't planned on testing the P. microspora until you get things figured out. Got too much else going on...

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Re: Pestalotiopsis microspora [Re: ghiajake]
    #22436759 - 10/26/15 04:42 PM (8 years, 6 months ago)

My P. microspora (from the Co-op buy) separated into two distinct looking cultures-- one i assume is a contaminant. Could I get some help on culture ID? Much Appreciated !



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Re: Pestalotiopsis microspora [Re: KingSloth]
    #22436857 - 10/26/15 05:16 PM (8 years, 6 months ago)

Mine is similar in appearance to the top photograph.

That being said the bottom photo is definitely the right one, the black spots are the spore masses.


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Re: Pestalotiopsis microspora [Re: KingSloth]
    #22437046 - 10/26/15 06:12 PM (8 years, 6 months ago)

Quote:

KingSloth said:
My P. microspora (from the Co-op buy) separated into two distinct looking cultures-- one i assume is a contaminant. Could I get some help on culture ID? Much Appreciated !




A good way to isolate it might be a non-nutritive polyurethane based medium like in the study:

Quote:

Initial PUR clearance screen.

Endophytes were first assayed for their ability to degrade PUR by growing them in the presence of Impranil DLF an anionic aliphatic aqueous PUR dispersion with 4% N-methyl pyrrolidone (NMP) (Bayer MaterialScience). Fifty-nine fungal endophytes were grown on solid PUR medium (PUR-A) containing 19 mM NaH2PO4, 33.5 mM K2HPO4, 7.6 mM (NH4)2SO4, 2.5 mM Na citrate, 250 μM MgSO4, 19 μM thiamine, 0.05% Casamino Acids, 147 μM FeCl3·6H2O, 14 μM ZnCl2·4H2O, 12 μM CoCl2·6H2O, 12 μM Na2MoO4·2H2O, 10 μM CaCl2·2H2O, 11 μM CuCl2, 12 μM MnCl2, 12 μM H3BO3, and 1.8 mM HCl. To 1 liter of this mixture was added 10 ml Impranil DLF and 15 g of agar. The polymer was added after autoclaving the medium to prevent deformation.




Impranil DLF Dispersion:

Quote:

Form supplied:
White low-viscosity dispersion in water with a solids content of
approx. 40 %; contains 4 % N-methyl pyrrolidone




https://www.pricefalls.com/product/n-methyl-2-pyrrolidinone-nmp-aka-n-methyl-pyrrolidone-paint-stripper-one-gallon/68815529

http://www.lowes.com/Stains-Sealers/Stain-Finishes-Sealers/Polyurethanes/_/N-1z0uavg/pl

I'm guessing the dipropylene glycol is the pylol and M2P counts as the isocyanate? (it's possible the electrons in the 5-member ring act aromatically which would account for the missing double-bond. i think they do on two of the sides). Regardless, seems like the same stuff:

http://www.rustoleum.com/MSDS/ENGLISH/260170.pdf

Quote:

Chemical Name                                      CAS-No.                Weight %
          Dipropylene Glycol Monomethyl Ether    34590-94-8          5.0
          Methyl-n 2-Pyrrolidone                          872-50-4              5.0




http://www.ncimb.com/Files/GrowthMediaRecipes.pdf


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OfflineMrcloudy
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Re: Pestalotiopsis microspora [Re: micro]
    #22437191 - 10/26/15 06:50 PM (8 years, 6 months ago)

I don't know if I posted it in this thread, but I am using Minwax Polycrylic, a water based polyurethane. I have not figured out exactly how to add it to the agar yet. I did learn that you can add it to the agar post sterilization without it being a significant contamination vector. But I still had separation. Perhaps the agar needs to cool further.

I saw no significant clearing of the agar in either of the three strains I used. I either used too much polyurethane, or it was because it broke up into chunks that were harder to digest than a fine dispersion. So my next round I will let the agar cool till it is quite thick, then add the polyurethane.


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MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.

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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22437341 - 10/26/15 07:24 PM (8 years, 6 months ago)

Hmm,

Maybe from the heating? How are you stirring it and for how long?

Also, how much is it diluted?

The methyl-n 2-pyrrolidone should be sol. in water but it seems the dipropylene glycol ether might not not: http://msdssearch.dow.com/PublishedLiteratureDOWCOM/dh_012d/0901b8038012d976.pdf?filepath=oxysolvents/pdfs/noreg/110-00977.pdf&fromPage=GetDoc

That's rather strange if it is water-based.

If that were the case, you could possibly get it to work by adding something like DMF.

Another option might be to simply buy the M2P and use the dimethyl ether, instead.


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Re: Pestalotiopsis microspora [Re: KingSloth]
    #22440437 - 10/27/15 02:09 PM (8 years, 6 months ago)

Quote:

KingSloth said:
My P. microspora (from the Co-op buy) separated into two distinct looking cultures-- one i assume is a contaminant. Could I get some help on culture ID? Much Appreciated !








My culture looks like neither of these...lol


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OfflineAlan RockefellerM
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Re: Pestalotiopsis microspora [Re: leschampignons]
    #22441957 - 10/27/15 08:26 PM (8 years, 6 months ago)

Quote:

leschampignons said:
My culture looks like neither of these...lol





What does your culture look like?

It should look like the bottom one.  I think the top dish in the above pic is some kind of contamination.

One fun thing to do is scope the black dots at 400x or 1000x, they look really neat.

I wonder if it actually eats plastic?  Has anyone tried this?

I am not sure which things have polyurethane in them.  I know they sell polyurethane condoms for people who are allergic to latex.  I wonder what else has polyurethane.  There is this:  https://en.wikipedia.org/wiki/List_of_polyurethane_applications.&nbsp; However I am not sure how to tell for sure that something is polyurethane and not some other plastic.

One possible thing to do is let a dish colonize, then cut a little square from a polyurethane condom and put it on the top of the dish and see if it gets eaten.

It would also be interesting to try some other plastics.

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Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22442079 - 10/27/15 08:48 PM (8 years, 6 months ago)

Quote:

Alan Rockefeller said:
I am not sure which things have polyurethane in them.  I know they sell polyurethane condoms for people who are allergic to latex.  I wonder what else has polyurethane.  There is this:  [url=https://en.wikipedia.org/wiki/List_of_polyurethane_applications. ]https://en.wikipedia.org/wiki/List_of_polyurethane_applications. [/url];  However I am not sure how to tell for sure that something is polyurethane and not some other plastic.

One possible thing to do is let a dish colonize, then cut a little square from a polyurethane condom and put it on the top of the dish and see if it gets eaten.

It would also be interesting to try some other plastics.




They used a water-based dispersion in the reference article.

I can't find out where to get Impranil DLF, but I posted a link for N-metyl-2-pyrollidone above.a

I also posted a link for polyurethane from Lowe's, but something like this might be better:

http://www.ebay.com/itm/like/310219352208?ul_noapp=true&chn=ps&lpid=82

You don't even need the lactone according to the article; the stuff grew off of only PUR.

The lactone will probably be digested by a number of things (I'm guessing a bit but it would obviously be easily oxidized and then probably broken down further).


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Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22443176 - 10/28/15 02:40 AM (8 years, 6 months ago)

Quote:

Alan Rockefeller said:I am not sure which things have polyurethane in them.  I know they sell polyurethane condoms for people who are allergic to latex.  I wonder what else has polyurethane.




Skateboard wheels are usually made from polyurethane. One wheel would provide plenty of material for testing. Just shave off thin slices.


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Re: Pestalotiopsis microspora [Re: BlimeyGrimey]
    #22443188 - 10/28/15 02:45 AM (8 years, 6 months ago)

The problem there... is no water.

Why not use a water-based dispersion?

You know, like... in the reference article :V


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OfflineKingSloth
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Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22447479 - 10/29/15 12:40 AM (8 years, 6 months ago)

Quote:

Alan Rockefeller said:
I wonder if it actually eats plastic?  Has anyone tried this?






I'm trying it with different concentrations of store bought water-based PUR on agar plates. It looks like it will grow in concentrations of about 12.5% but not 25%...so somewhere in there is the cutoff point. As for it's plastic eating qualities the article says that it will show a "zone of clearance" -- turning the mixture from opaque to clear around the growth area. It looks like this might be happening on the right plate, but its hard to tell... I think what i'm going to do next is try it out with a control fungus so I can compare... and maybe try mixing the PUR into the Agar.

Also, I've I've done some trials with jars of garden soil and it looks like it will establish...so it potentially has the ability to clean up PUR residues in soils!




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Re: Pestalotiopsis microspora [Re: KingSloth]
    #22447500 - 10/29/15 12:51 AM (8 years, 6 months ago)

I have been trying it with the water based polyurethane as well, ran three plates each of my Pestalotiopsis cultures.

None showed any clearance zone for me not even the ATCC group buy culture, I also still get a small amount of separation with the polyurethane leaving little white specks in the agar. Not as bad as when I full on pressure cooked it though. Not going to bother posting photos because the cultures have filled the plates now, nothing to see anyways. I will continue some more.

The good news is though that if your culture produces those little black liquid balls... you now have tons of spores to make more cultures with and see if you can't find one that does a better job at eating the plastic than the parent culture.


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AMU

MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.

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Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22447511 - 10/29/15 12:56 AM (8 years, 6 months ago)

Quote:

Alan Rockefeller said:


One fun thing to do is scope the black dots at 400x or 1000x, they look really neat.






I second this, these species have some of the most interesting spores I have seen yet.

From my culture isolated from Alabama.


From my culture isolated from Florida


I don't have a photo of the ATCC culture but it looks pretty much exactly like my Florida spores. The Alabama spores are larger (The images above are not to scale though)  and have a greenish hue.


--------------------

10 different Ganoderma species from across the USA

AMU

MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.

Edited by Mrcloudy (10/29/15 12:56 AM)

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Re: Pestalotiopsis microspora [Re: leschampignons]
    #22447978 - 10/29/15 06:42 AM (8 years, 6 months ago)

Quote:

leschampignons said:
My culture looks like neither of these...lol



probably just because it is on activated charcoal agar


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Re: Pestalotiopsis microspora [Re: nomendubium]
    #22449895 - 10/29/15 04:32 PM (8 years, 6 months ago)

activated charcoal? o.O

... what is trying to be absorbed?


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Re: Pestalotiopsis microspora [Re: nomendubium]
    #22455734 - 10/30/15 09:49 PM (8 years, 6 months ago)

I def see the black dots on the one you sent me but I don't think I see any on the transfers I did. How long do they take to appear? My transfers definitely show filamentous white growth with maybe some clear droplets on top? I can try to upload a photo but I use jars for petris so it's hard to get a good shot

Edit: I oughta scope em too to check


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Edited by leschampignons (10/30/15 09:50 PM)

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Re: Pestalotiopsis microspora [Re: leschampignons]
    #22470067 - 11/03/15 08:51 AM (8 years, 6 months ago)

I didn't keep track, but not long, maybe 2 weeks. The charcoal wasn't really for anything, i just had some extra dishes of it left over from some side by side comparisons. In my tests, the MEA with charcoal grew faster than regular MEA with 2 different oysters, Cyclocybe aegerita and the present species. H. erinceus was a little slower, initially taking over a week to jump on to the agar. When it finally did it grew at least as fast though


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Re: Pestalotiopsis microspora [Re: nomendubium]
    #22470142 - 11/03/15 09:16 AM (8 years, 6 months ago)

huh, weird... generally it is to absorb some unwanted compound


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