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Amazon Shop for: Agar, Autoclave, HEPA Filter, Laminar Flow Hood, Paul Stamets, Petri Dish, Pressure Cooker, Rye Grain

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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73307 - 01/30/00 10:09 PM (16 years, 10 months ago)

i think you spilled cheese puffs in your agar


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Anonymous

Re: Contaminated Agar...what is this shit! (pics) * 1
    #73309 - 01/30/00 10:51 PM (16 years, 10 months ago)

The yellow blotch I have seen before. It's pretty common, smells like rotting apples. You must have some problem with something, and my guess is media. How long do you sterlize your agar for? You should do it for 30-40 mins, in a flask with a cotton swab stuffed in the neck and tin foil on top. You should let you pressure cooker cool down normally and then put a paper towel soaked with alcohol or Lysol over the stopcock/neck to prevent contaminents from entering as it cools. Other than that, just wipe your tools, surfaces and gloves with alcohol or 15% bleach/water mix.


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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73310 - 01/31/00 05:32 AM (16 years, 10 months ago)

Those are bacterial contams
I have trouble with those.

1. Sterilise the media properly
2. Alcohol isn't good enough for bacteria
3. Use Lysol

No 1 is the most important since depending on the media some pretty heat resistant bact. contams exist.

Do many dishes and that way u are assured of a few clean ones.



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InvisibleRyche HawkV
A Messenger

Registered: 03/01/01
Posts: 2,062
Re: Contaminated Agar...what is this shit! (pics)
    #73311 - 02/04/00 01:09 PM (16 years, 9 months ago)

I run into this problem sometimes as well,, and I use a very large high dollar flow hood.

I find this problem ususally occurs from condenstaion on the top of the agar plate, the water droplets drop on the agar and usually start this.

From my recent experiments with agar, I always wipe all the dishes down with alcohol first, let them dry of course. I wait as long as possible before pouring my agar, when its cool enough to barely leave condensation, but also still warm enough that its still thin to pour. When some dishes still condensate, I go back with a lint free cloth, presoaked in alchol, then allowed to dry, then I dry the top of the petri dish lids. This has cut down my contam rates with petri dishes considerably. Its a bit of xtra work but has paid off.

-peace-



--------------------
-Peace-
www.MushroomS.com
Formerly
www.thehawkseye.com
Sacred Mushroom Spores


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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73312 - 02/04/00 02:26 PM (16 years, 9 months ago)

Does adding gentamycin (an anti-biotic additive) to agar ala Smart Botanics work well?


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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73313 - 02/04/00 03:25 PM (16 years, 9 months ago)

That's an intersting idea with the drying Ryche Hawk. I don't have a flow hood, but I did a lot of investigation on this condensation issue, and here's what i have found.

Stacking your dishes helps a hell of a lot. The uppermost dish (heat rises of course) will have the worst effects, so just sacrifice this one. Also leaving warm water in the top dish helps too. I usually incubate my petris upside down, this is a fairly common lab practise and helps a lot.
Also like everyone has said, leaving the agar untill it is nearly gelled before pouring will help.



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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73314 - 02/04/00 03:47 PM (16 years, 9 months ago)

I defintely remember some post about 3 months ago concerning using Peroxide in agar. I tried the search engine but gave up after 5 mins. of waiting. Good luck!


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Invisibleshane67
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Registered: 12/01/00
Posts: 113
Re: Contaminated Agar...what is this shit! (pics)
    #73315 - 02/04/00 04:26 PM (16 years, 9 months ago)

Thanks to everyone that responded to my post.

Ryche Hawk...that is it!

A lot of water has gotten into my petri dishes. I also have been pouring the Agar when it is just cool enough to handle (very hot). I will try again tonight (just recently got a flask w/glass stopper).

I am going to pick-up some yeast for a mycelium brew tonight?wish me luck!



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InvisibleRyche HawkV
A Messenger

Registered: 03/01/01
Posts: 2,062
Re: Contaminated Agar...what is this shit! (pics)
    #73316 - 02/05/00 01:54 AM (16 years, 9 months ago)

Yeah.... I've noticed a tremendous decrease in contaminated dishes... especially with this yellow cheesy looking mold.. since I cut down the condensation in the agar dishes.

And as DirtyDog mentioned.... I stack mine as well .. and only the top ones condensate and I have to retry the first 3 or 4 in a stack of 10 using my new mentioned methods above.
DirtyDog.. you also mention storing them upside down... are you talking about AFTER the mycelium is growing?
I dont have much problems with them later.
Because I look real close at the dishes before I even inoculate. If I see any dishes with those little yellow blemishes or cheesy or any other kind of suspicous mold growing.. i just throw them out now. They'll contaminate later.. so why bother.

No joke... i use to loose at leat 1/2 of my dishes to this cheesy mold before I relized it was being caused by the condensation.
Yet.. when heavy mycelium is growing... and condenstation develops later and drops on they mycelium.. it doenst seem to hurt anything. The mycelium just absorbs it... as it does when its in cake or casing form.

Loosing 1/2 of my dishes turned me off from agar work for a long time.... its hard enough work as it is... only to throw away 1/2 of waht you made. Try my suggestion above...it should help with some of these common petri dish molds.

StarRover,
I have never used the anti-biotic in my petri dishes. It supposedly damages the mycelium and can weaken the strain. And I've read lots of reports of ppl still haveing major contam problems even when they used it.
If my memory serves me correct, Stamets only recommends using it when one is haveing lots of problems with contams on agar regularly.
But if used continasly, it weakens the strain.

I use the agar sold by Fungi Perfecti, without the anti-biotic, and since I've eliminated this condensation problem, I only loose about 1 out of 12 dishes or so to agar contams.... much better then 1/2.

Much happier now and actually using agar again (but only when i have to.. still hate the suff.. multi-spore inoculation rocks !!! )

-peace-



--------------------
-Peace-
www.MushroomS.com
Formerly
www.thehawkseye.com
Sacred Mushroom Spores


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Invisibleshane67
Keeper
Registered: 12/01/00
Posts: 113
Re: Contaminated Agar...what is this shit! (pics)
    #73317 - 02/05/00 09:55 AM (16 years, 9 months ago)

Guys & Gals

When I first started growing I had almost 100% non-contamination rate (glove box & foil to wrap my dishes). Now that I have a flow hood I feel I didn't need to use foil (which is rightly so). Well, I am going back to the foil. I recall no extra moisture build-up after autoclave and had great results.



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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73318 - 02/06/00 12:47 AM (16 years, 9 months ago)

What kind of HEPA are you using? is htis the furnace HEPA or the LAB HEPA?


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Invisibleshane67
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Registered: 12/01/00
Posts: 113
Re: Contaminated Agar...what is this shit! (pics)
    #73319 - 02/09/00 02:12 PM (16 years, 9 months ago)

Gainen,

I am using the Lab HEPA. A matter-a-fact I wrapped my petri dishes in foil and allowed the Agar to cool...not a single contamination! Thank you Ryche Hawk and everyone that helped me solve my problem!



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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73320 - 02/09/00 03:57 PM (16 years, 9 months ago)

Good ideas Rhyche. I am having some serious probs with contams right now. I don't have any good prints either which doesn't help (ordered one from you though). I might try using the tin foil, I was using plastic wrap but it doesn't seem to be doing the job. I agree with Rhyche though, for some reason once you have mycelium condensation doesn't seem to be a problem. I am getting to the point where I might just do a multi spore into rye just to get a healthy mycelium. ARG!


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Anonymous

Re: Contaminated Agar...what is this shit! (pics)
    #73321 - 02/09/00 03:59 PM (16 years, 9 months ago)

Just noticed that I spelled Ryche wrong. : )


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InvisibleRyche HawkV
A Messenger

Registered: 03/01/01
Posts: 2,062
Re: Contaminated Agar...what is this shit! (pics)
    #73322 - 02/12/00 11:08 PM (16 years, 9 months ago)

You guys are using tin foil to wrap your dishes. Let me save you a lot of time.
Spend the $35 for a roll of para film from fungi perfecti. This stretchy plastic ROCKS !!!! And it goes a long long way, that roll will last you for years.

-peace-



--------------------
-Peace-
www.MushroomS.com
Formerly
www.thehawkseye.com
Sacred Mushroom Spores


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Amazon Shop for: Agar, Autoclave, HEPA Filter, Laminar Flow Hood, Paul Stamets, Petri Dish, Pressure Cooker, Rye Grain

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