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OfflineMushDog
A Dog's Boy
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Registered: 03/30/11
Posts: 5
Last seen: 12 years, 8 months
Liquid culture question - Contaminants and the prevention of
    #14405964 - 05/05/11 03:57 PM (12 years, 8 months ago)

Alright, ladies and gents. I'm going to Hulk-out and smash my kitchen to bits if I can't figure something out.

I will provide some information on my situation, then I will pose a question. If you have no answer to the question, or you have the answer I'm afraid I'm going to get (which is, 'Yes'), then I'd be happy to get some further advice on the topic.

Now, then, brass tacks.

I had three perfect, little cakes (I thought they were, anyway). I lost two to a vicious and nasty green mold. I quarantined the third, but it was too late.

I then started over with a liquid culture of 100cc water, 3cc honey, and 2cc spores. It grew faster than I thought it would. In less than two days, I had a jar of the foulest smelling mold I've ever put my nose near.
I repeated the process, this time making double-sure I prevented contamination. I boiled the CRAP out of my equipment. I still lost my second culture (same ratios) to mold.
I have settled on the conclusion that I must have contaminated my spore syringe. I have no prints.

The only specimen I have to start another culture now is some of my prized, and now rare, dried babies (I have less than a gram). I have no money that I can spare on this (especially not when considering shipping costs), and I don't expect my wife to start crapping diamonds anytime soon.

Now, I've been made to understand that I can do a LC with fresh or dried mushies. I could start, yet again, with my coveted dried samples, but I have one thing stopping me: No H2O2. This doesn't sound like much, but my samples have been sitting in a cupboard for several days, and I just can't be sure they aren't covered with something sinister.
Finally, my question:
If I boil my mushrooms to assure sanitation, will the samples become useless for the purpose of starting a liquid culture?

Of course, if you have any other remedies to my problem that I've overlooked, I will give the elderly woman living next door to me a night to remember while she lays on her deathbed, with a ghost of a smile haunting her face.


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Note: MushDog is a dog with a terminal case of tomfoolery. No authority, agency, or entity should take the above post as truth. It is, actually, a work of fiction by a mentally inept individual, who has done zero of the things he claims to have done.


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Invisiblewow
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Registered: 07/27/06
Posts: 1,260
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Re: Liquid culture question - Contaminants and the prevention of [Re: MushDog]
    #14405997 - 05/05/11 04:03 PM (12 years, 8 months ago)

starting LC's from spores has always been a crapshoot for me :shrug:  i always use a clone or small piece of agar and usually never use the lc more than once or twice..  idk just a me thing...  it would be a long shot to get a culture started from boiled shrooms actually, yea youll kill anything with hopes to grow by boiling..  if you have agar however you can sometimes revive some inner stem tissue on the agar..


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InvisibleTheMadSeasonV
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Registered: 05/16/05
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Re: Liquid culture question - Contaminants and the prevention of [Re: MushDog]
    #14406019 - 05/05/11 04:09 PM (12 years, 8 months ago)

You need to hydrate the tissue with water and put it on agar. A liquid culture is doomed to failure with dried mushroom tissue unless you somehow picked and dried those mushrooms in sterile conditions, which is very unlikely.

-Mike


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Invisiblebbox244
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Registered: 03/17/11
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Re: Liquid culture question - Contaminants and the prevention of [Re: wow]
    #14406073 - 05/05/11 04:17 PM (12 years, 8 months ago)

On the last contaminated jar, was the entire jar mold?  Is there a chance you could pull of a piece of myc. and do an agar isolate, may take a few isolates, but would this work? Guys, I'm kind of new to agar, but if he sections properly and takes multiple samples wouldn't he inevitably get an isolate of quality for which to make an LC with?  I doubt you'll get anything from dried shrooms, unless they are moist, then you could do a center stem sample.  But you said "dried", and I don't think you can get anything from dried shrooms.  Did you ditch the third jar?  Is it completely fried? Maybe dried shrooms work, that part I'm unsure of, someone more experienced will have to comment on that, but as far as boiling, it kills myc. and spores. So, you can't do that.


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OfflineMushDog
A Dog's Boy
Male
Registered: 03/30/11
Posts: 5
Last seen: 12 years, 8 months
Re: Liquid culture question - Contaminants and the prevention of [Re: TheMadSeason]
    #14406114 - 05/05/11 04:23 PM (12 years, 8 months ago)

Do not boil- roger wilco, Joe.

Now about agar... As I mentioned, I am pitifully poor. I've seen some 'poor man' recipes for agar involving honey, potato flakes, and water. Are these recipes any good?

And what's preventing these same contaminants from growing in the agar? Do I just have to take my sample before the contams get a chance to start?

bbox244 - I still have my very, very moldy LC. I've still got it purely because I'm afraid of smelling it again. When I looked, it seemed it was mostly green mold, but there may have been a bit of mycelium. The entire solution is tinted green, so it's pretty hard to spot something white in it.


EDIT:Mother-Superior jumped the gun! Hold the phones, folks!
I just realised a very basic mistake I made. I've been inspecting my jars in low-light. I checked it out in better lighting, and it's not as disgusting as I thought it was. It still has a very potent and offensive odor, however. It smells like something I picked out of a large, full trash can. But there is definitely spots of healthy-seeming mycelium.


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Note: MushDog is a dog with a terminal case of tomfoolery. No authority, agency, or entity should take the above post as truth. It is, actually, a work of fiction by a mentally inept individual, who has done zero of the things he claims to have done.


Edited by MushDog (05/05/11 04:31 PM)


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Invisiblewow
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Registered: 07/27/06
Posts: 1,260
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Re: Liquid culture question - Contaminants and the prevention of [Re: MushDog]
    #14406212 - 05/05/11 04:42 PM (12 years, 8 months ago)

its gonna be hard to isolate a culture from that being there is mold and contam spores all through it..  your gonna contaminate your work space most likely..  poor man agar isnt gonna work well..


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Invisiblewow
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Registered: 07/27/06
Posts: 1,260
Loc: Cloud 9
Re: Liquid culture question - Contaminants and the prevention of [Re: wow]
    #14406216 - 05/05/11 04:43 PM (12 years, 8 months ago)

you know what man...  im gonna help you out..  its your lucky day possibly...:thumbup:


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OfflineMushDog
A Dog's Boy
Male
Registered: 03/30/11
Posts: 5
Last seen: 12 years, 8 months
Re: Liquid culture question - Contaminants and the prevention of [Re: wow]
    #14406253 - 05/05/11 04:52 PM (12 years, 8 months ago)

Y'know... You guys are my white knights. After inspecting my jars in better lighting, I think I might actually have an uncontaminated jar of LC. I'll be monitoring it VERY closely for the next couple days, and this weekend I'll be testing it. Thanks for all the help, guys! I might actually start up a journal later tonight to chronicle my habits of suspecting the worst.


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Note: MushDog is a dog with a terminal case of tomfoolery. No authority, agency, or entity should take the above post as truth. It is, actually, a work of fiction by a mentally inept individual, who has done zero of the things he claims to have done.


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Invisiblewow
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Male User Gallery


Registered: 07/27/06
Posts: 1,260
Loc: Cloud 9
Re: Liquid culture question - Contaminants and the prevention of [Re: MushDog]
    #14406322 - 05/05/11 05:07 PM (12 years, 8 months ago)

a good way to tell is; if it has globs of myc and the liquid is clear as it was when you started its probably good,  if its cloudy or has dust lookin clouds starting to form its definitely contammed!  check your pms mushdog!


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