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InvisibleYidakiMan
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black pin mold on peroxidated MEYA
    #1851890 - 08/26/03 05:37 PM (13 years, 5 months ago)

A dozen petris had slices of mycelium transferred onto them. A day later they had all lept off. Another day later over half had gotton black pin mold. Always seal the edges of your petris.


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OfflinePaid
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1852102 - 08/26/03 06:35 PM (13 years, 5 months ago)

Agreed, sealing = key.
Choose the correct sealing tape as well.
For me cling film (saran wrap(sp?) works best.
I have had complete faliures with selotape and marsking tape
:-(
What taps or seals have others used to success or failiure?

?


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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: Paid]
    #1852536 - 08/26/03 08:50 PM (13 years, 5 months ago)

The remainder I sealed up with regular cellophane tape. I really wonder if they are still stuck.


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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1856472 - 08/27/03 10:42 PM (13 years, 5 months ago)

Three more contams. Two black pin, the other is a speckling of some type of white mold. How many different ways can you say shit?


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OfflineTheSwazi
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1863319 - 08/29/03 06:25 PM (13 years, 5 months ago)

okay the speckling white mold might just be the pinning of the new mushrooms. The black pins are definitely a mold of the top of the pins. I wouldn't worry too much about the white stuff though.


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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: TheSwazi]
    #1863429 - 08/29/03 07:11 PM (13 years, 5 months ago)

I took transfers anyway. The new dishes are all wrapped in saran wrap.


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OfflineMAIA
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1870720 - 09/01/03 10:21 AM (13 years, 5 months ago)

Where and how did you make the transfer ?
That's a key point. You should be using a glove box or a HEPA equipped room for such procedure.

MAIA


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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: MAIA]
    #1871437 - 09/01/03 04:27 PM (13 years, 5 months ago)

Well this is interesting. That last batch of agar I accidentally doubled the peroxide. It was 10ml peroxide for 500ml of agar. Immediately after transfer each dish was wrapped in saran wrap. All but 2 contamed with black pin mold. Each black pin bloom is eminating from the edge. Neither of my two blanks contamed.


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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1874444 - 09/02/03 04:37 PM (13 years, 5 months ago)

Any body got any idea on how to stop this? I have lost so many dishes my hopes of isolating a strain is basically cached. I have lost 75% of 30 dishes. Each contam is black pin mold eminating from the edge of the dish.

Here's my procedure:

1. Prepared 500ml MEYA using the following recipe:
10g agar
10g malt extract
2 grams yeast.

2. Sterilized for 40 minutes

3. Moved to a cleaner room than a kitchen, partly opened to allow thermometer.

4. Cools to 130 F. and 10ml H2O2 added.

5. In the cleaner room, agar is pored into reused plastic petris. Allowed to cool.

6. Same room, established cultures are cut with a flame sterilized knife and moved onto new agar plate.

7. Petri is sealed with Saran wrap and placed in incubator.


Do I have to pour and inoculate peroxidated agar in my glove box? I tried it once, it was a disaster! The bottom of my glovebox is not flat. It is difficult to say the least.


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OfflineMAIA
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1877350 - 09/03/03 02:24 PM (13 years, 5 months ago)

When the Petris are opened, they should be opened inside the glove box, anything called "cleaner room than a kitchen" is no good for agar work. Are you using lids for the Petris ? How and where do you place the lids when you open the Petri ? For how long are the Petris exposed to open air ? This procedures all also very important.
Other problem could be the mycelium, even if you use H2O2, some molds can survive it if the mycelium is infected.
My recomendation is to do everything inside the glove box. Clean every material going inside with a towel soaked with alcohol and do your stuff.

MAIA


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Use, do not abuse; neither abstinence nor excess ever renders man happy.
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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: MAIA]
    #1877815 - 09/03/03 04:26 PM (13 years, 5 months ago)

Thanks for your help! I used the Peroxidated Agar Tek available here at the shroomery. IMHO, it misleads the reader to believe that aseptic technique is not necessary.


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Offlinebonsai
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Re: black pin mold on peroxidated MEYA [Re: YidakiMan]
    #1916992 - 09/15/03 01:15 AM (13 years, 5 months ago)

I had the same problems that you are talking about and the glove box with the rounded bottoms doesn't help  :frown:.  Everything now is being done in front of a laminar flow box.  So far no contams.  So how you do it and where are major issues.  Even in the lysoled glove box I would pick up contams.  Where they come from is beyond me.  So far with the HEPA flow box everything is contam free.  I seal the sides with Parafilm.
bonsai   


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InvisibleYidakiMan
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Re: black pin mold on peroxidated MEYA [Re: bonsai]
    #1919872 - 09/15/03 11:41 PM (13 years, 5 months ago)

I am just about to finish my flow hood. =)


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