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OfflineSharpstuff


Registered: 05/04/11
Posts: 510
Loc: Australia Flag
Last seen: 7 years, 10 months
Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22173040 - 09/01/15 08:51 AM (8 years, 8 months ago)

Yeh or a wedge would be great if anyone has one. Surely there are some auzzies with this culture?

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Offlinegreenelephant
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Registered: 10/31/12
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Last seen: 6 years, 6 months
Re: Pestalotiopsis microspora [Re: Sharpstuff]
    #22177027 - 09/02/15 04:50 AM (8 years, 8 months ago)

Hello. Does anyone still have a sample of pestalotiopsis microspora available to send to the uk? will negotiate the necessary prerequisites. thanks :smile:

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OfflineButhoscorpio
Registered: 01/05/11
Posts: 96
Last seen: 2 years, 8 months
Re: Pestalotiopsis microspora [Re: greenelephant]
    #22177121 - 09/02/15 06:04 AM (8 years, 8 months ago)

I sent you a message. I can send the cultures to european mycologists.

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Offlineaktar
nerf herder

Registered: 02/27/11
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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22194039 - 09/05/15 06:56 PM (8 years, 8 months ago)

Quote:

Mrcloudy said:
Water based polyurethane, it must be added after sterilization. If you mix it in before pressurecooking it precipitates out into strings and chunks of plastic. I was able to confirm that there does not seem to be anything in the water based polyurethane that inhibits the fungal growth, though I did confirm that my strain will not grow on straight polyurethane agar alone. 

The water based polyurethane is opaque, you can see the fungus eating it because it will clear up around the mycelium on the plate.

I will be continuing my experiments here soon, I will try sterilizing the polyurethane separately then adding it, if it can survive pressure cooking on its own.





This is cool. Keep us posted :thumbup:

Where did you get water-based polyurethane? is it in solution? I'm interested in this

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Offlinebeendobarna
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Registered: 09/14/15
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Re: Pestalotiopsis microspora [Re: aktar]
    #22238303 - 09/14/15 07:42 PM (8 years, 8 months ago)

Hi I'm now of the donators who never received anything.
Are there any cultures available still?

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OfflineMrcloudy
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Re: Pestalotiopsis microspora [Re: aktar]
    #22238591 - 09/14/15 08:50 PM (8 years, 8 months ago)

Quote:

aktar said:



Where did you get water-based polyurethane? is it in solution? I'm interested in this







Basically this, it is polyurethane held in suspension, it is opaque due to all the small particles, which is the key to its use to screen for plastic eating abilities. It makes the agar cloudy, and if the fungus can eat plastic the agar will clear up around the mycelium.

I have tested it and there seems to be nothing in it that inhibits fungal growth, which I was concerned about because it is not entirely pure water and polyurethane, there are a few added solvents. But there was no problem with fungal growth.

My main goal for it at this time is figuring out how to incorporate it into the agar, cannot be added prior to pressure cooking.

I have been busy with other projects and haven't gotten to the polyurethane agar yet. But am getting there. Got a nice new flowhood set up so I can do a lot more agar work.


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

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.

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Offlineforrest
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Registered: 11/16/12
Posts: 1,011
Loc: The Netherlands
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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22251465 - 09/17/15 02:22 PM (8 years, 7 months ago)

that's a great idea! what if you first let it colonise the plate, and then just drip some paint on the myc?


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My Trade List

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Offlinenomendubium
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Registered: 05/16/14
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Re: Pestalotiopsis microspora [Re: beendobarna] * 2
    #22278141 - 09/23/15 10:25 AM (8 years, 7 months ago)

Quote:

beendobarna said:
Hi I'm now of the donators who never received anything.
Are there any cultures available still?



I still have some. Pm me the name you used when you contributed and I will check the list and send you one if you are in the US, if you are in Another country, I will refer you to the right person


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OfflineAlan RockefellerM
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Re: Pestalotiopsis microspora [Re: nomendubium]
    #22278215 - 09/23/15 10:48 AM (8 years, 7 months ago)

Quote:

nomendubium said:
I still have some. Pm me the name you used when you contributed and I will check the list and send you one if you are in the US, if you are in Another country, I will refer you to the right person





You are doing great work!

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Offlinepintas
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Registered: 09/24/15
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Last seen: 8 years, 7 months
Re: Pestalotiopsis microspora [Re: nomendubium]
    #22287400 - 09/25/15 07:51 AM (8 years, 7 months ago)

Quote:


I still have some. Pm me the name you used when you contributed and I will check the list and send you one if you are in the US, if you are in Another country, I will refer you to the right person




I did not contribute (didn't know about it then), but i really need some... Sent you a PM.

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OfflineSharpstuff


Registered: 05/04/11
Posts: 510
Loc: Australia Flag
Last seen: 7 years, 10 months
Re: Pestalotiopsis microspora [Re: nomendubium]
    #22326636 - 10/03/15 01:18 AM (8 years, 7 months ago)

Quote:

beendobarna said:

I still have some. Pm me the name you used when you contributed and I will check the list and send you one if you are in the US, if you are in Another country, I will refer you to the right person




Will you send a culture to Australia?

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OfflineAlan RockefellerM
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Re: Pestalotiopsis microspora [Re: Sharpstuff]
    #22327967 - 10/03/15 12:25 PM (8 years, 7 months ago)

Cultures are really hard to get into Australia - a really thin agar wedge might make it, but it is likely to be intercepted and burned if the envelope is significantly bumpy.

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Offlinemorbiddoctor
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Registered: 07/05/09
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Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22328088 - 10/03/15 12:56 PM (8 years, 7 months ago)

Just got confirmation that I'm getting my hood back! Looking forward to working with this fungus. If anyone can spare a wedge I'd be forever grateful.


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I'm looking for eyecatching, colorful, unique and interesting fungi. Not limited to edible or medicinal. Print them when you're on foray and pm me please!

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Invisiblemicro
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Registered: 05/09/03
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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22330988 - 10/04/15 12:18 AM (8 years, 7 months ago)

Quote:

Mrcloudy said:
My main goal for it at this time is figuring out how to incorporate it into the agar, cannot be added prior to pressure cooking.




Here is a reference, in case it helps:

Biodegradation of Polyester Polyurethane by Endophytic Fungi

Quote:

Initial PUR clearance screen.

Impranil DLN, a polyester polyurethane (PUR), is an opaque milky suspension that becomes transparent upon degradation. Organisms capable of degrading this polymer display a zone of clearance around the growing culture (5). A collection of 59 fungal endophytic organisms isolated from plant samples in the Ecuadorian Amazon were screened for their ability to grow on and degrade polyester polyurethane using the PUR halo assay as the initial screen. Of the organisms screened, 18 organisms produced a halo of clearance such as that shown in Fig. 1. Two other organisms, identified by ITS sequencing as Guignardia mangiferae (E2702C) and Zopfiella karachiensis (E2719A), could grow on but not degrade PUR-A medium. These were used as negative controls in the subsequent studies. The host plants, isolation media, and identities of the 18 active fungi and two inactive control fungi are listed in Table 1.




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.

The solid medium screening assay followed the general method of Crabbe et al. (5). The PUR-A solid medium was added to sterile culture tubes in 10-ml aliquots. A 0.5-cm3 plug of fungus grown on PDA was added to each test tube using aseptic technique and allowed to grow undisturbed at 23°C. The bacterium Pseudomonas chlororaphis (ATCC 55729, from Gary Howard) and the fungus Aspergillus niger (from Gary Strobel, Montana State University) were used as positive controls for polyurethane degradation (6, 7, 13). PUR degradation was evidenced by a change in medium appearance from opaque to translucent. After 2 weeks of incubation, the depth of polyurethane clearance was measured from the top of the medium to the lowest point of visible clearance.




Quote:

Organisms identified as having PUR-degrading activity were tested for their ability to use PUR as the sole carbon source. For these studies, the substrate was Impranil DLN, which contains PUR suspended in only water (no N-methyl pyrrolidone is present). This isolates the Impranil as the sole source of carbon for metabolism and growth. The top five organisms from the initial activity screens were grown on Impranil DLN with no other carbon sources (PUR-Lmin). The fungal samples were washed prior to inoculation to remove all residual medium. These two considerations—the wash and the DLN substrate—ensure that the polymer is the sole source of carbon for fungal metabolism and growth. PUR-Lmin was prepared in a manner similar to that for the PUR-L medium but in the absence of sodium citrate, thiamine, Casamino Acids, or agar. The organism Aspergillus niger was tested as a basis for comparison.

Fungal cultures were grown for 1 week in potato dextrose broth (PDB). Stock cultures were homogenized by vigorous shaking, and 1 ml of each culture was centrifuged at 12,100 × g for 1 min. The supernatant was removed, and the fungal pellet was resuspended in 1 ml of the PUR-Lmin liquid medium. Samples were centrifuged and resuspended a second time to ensure removal of all residual PDB. The 1-ml sample of washed fungal material was added to sterile culture tubes to a final volume of 10 ml PUR-Lmin. The cultures were monitored for visual clearance of the opaque medium. Samples were measured every 2 days for 2 weeks for optical absorbance at 600 nm to determine an approximate rate of clearance. An increase in fungal mass correlating to PUR degradation was measured by lyophilizing mycelial mass from triplicate cultures containing minimal medium with and without PUR.






In case anyone is curious, here is the table with the positive fungi identified:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3165411/table/T1/


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Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)

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OfflineMrcloudy
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Re: Pestalotiopsis microspora [Re: micro]
    #22331790 - 10/04/15 08:53 AM (8 years, 7 months ago)

Quote:

micro said:


Here is a reference, in case it helps:

Biodegradation of Polyester Polyurethane by Endophytic Fungi

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.
[/url]





This is actually the study I used, the water based polyurethane from the shop is essentially what they used, but they used a more pure lab grade form that lacks one or two of the added solvents. I apparently totally missed that last bit there where it was added after autoclaving and learned that little bit on my own. :facepalm:


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

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.

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OfflineMrcloudy
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Re: Pestalotiopsis microspora [Re: Mrcloudy] * 1
    #22331802 - 10/04/15 08:55 AM (8 years, 7 months ago)

I also recently acquired as a contaminant yet another Pestalotiopsis culture, this one is interesting because it has greenish spores under the microscope and looks slightly different than the P.microspora. I will try all three cultures on the special agar soon.




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

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.

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OfflineAlan RockefellerM
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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22332083 - 10/04/15 10:29 AM (8 years, 7 months ago)

Cool!  Apparently the only way to even get to genus with this group is DNA sequencing.  If you have an extra $20 you can spare, send it to http://alvalab.es and ask for the ITS region.  Or ITS+LSU if you have $30.

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OfflineMrcloudy
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Re: Pestalotiopsis microspora [Re: Alan Rockefeller]
    #22340939 - 10/06/15 12:32 PM (8 years, 7 months ago)

Just made up 9 test plates. I think I added a bit more polyurethane than the recipe called for. 3 plates of Pestalotiopsis sensu lato from Floria, 3 plates of Pestalotiopsis sensu lato from Alabama, and 3 plates of the ATCC Pestalotiopsis microspora from the co op buy.

Here's to hoping that adding the plastic post sterilization did not add a contamination.  :toast:


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

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.

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Invisiblemicro
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Re: Pestalotiopsis microspora [Re: Mrcloudy]
    #22340974 - 10/06/15 12:48 PM (8 years, 7 months ago)

If you are using the media from that journal article it looks really minimal.

Your chances of growing anything else on it would be pretty slim.


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)

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OfflineMrcloudy
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Re: Pestalotiopsis microspora [Re: micro]
    #22340991 - 10/06/15 12:52 PM (8 years, 7 months ago)

I am only using the study as a guide for how much plastic to use the rest of the agar is typical MEA.


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

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.

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