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Offlineflipsidetrue
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Petri contams? Updated 11/06/13
    #19054643 - 10/29/13 10:20 PM (10 years, 3 months ago)

is it normal for mycelium to look this hairy and is it worth using?




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Edited by flipsidetrue (11/06/13 12:43 PM)


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OfflineEveniftheskyfalls
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Re: hairy mycelium? [Re: flipsidetrue]
    #19054676 - 10/29/13 10:24 PM (10 years, 3 months ago)

That looks like cobweb to me, but im a noob so dont take it to heart


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Re: hairy mycelium? [Re: flipsidetrue]
    #19054682 - 10/29/13 10:26 PM (10 years, 3 months ago)

It looks like the pictures of cobweb mold I've seen. How long did it take to grow that much? Cobweb will grow a lot faster and could take over a petri dish in a few days.


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Re: hairy mycelium? [Re: Stupendous-Yappi]
    #19054761 - 10/29/13 10:39 PM (10 years, 3 months ago)

Looks like cobweb to me as well. Tough break.


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Re: hairy mycelium? [Re: Giggle_Grower]
    #19054788 - 10/29/13 10:44 PM (10 years, 3 months ago)

Looks like you are performing a recrystalization.. And mycelium should not look like a recrystalization ha.


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Re: hairy mycelium? [Re: flipsidetrue]
    #19054840 - 10/29/13 10:55 PM (10 years, 3 months ago)

hm.  I'm a bit of a noob, but i've had a run-in with cobweb...  it didn't get that tall nor was it that white.  Luckily cobweb is the only contam you can really take control of.  Spray some peroxide on it.  If it melts, it's cobweb and keep spraying regularly throughout the day until it's gone...  If not, it's just the branchy kind of mycelium.


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Re: hairy mycelium? [Re: flipsidetrue]
    #19055267 - 10/30/13 12:07 AM (10 years, 3 months ago)

This is 100% definitely cobweb, and no, it is not worth using :sad:

garbage time!


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Re: hairy mycelium? [Re: PsiLisaBin]
    #19056617 - 10/30/13 08:58 AM (10 years, 3 months ago)

dat mold:cookiemonster:


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Re: hairy mycelium? [Re: PirateSwazey]
    #19057019 - 10/30/13 11:03 AM (10 years, 3 months ago)



Looks like cobweb to me.

Here's some cobweb I cultivated for reference.


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Re: hairy mycelium? [Re: bodhisatta]
    #19060354 - 10/30/13 09:22 PM (10 years, 2 months ago)

thanks guys i needed some back up opinions you all know how it is

thanks for the reference bodhisatta.

i went ahead and threw some fresh tissue on some agar, its been almost 48 hours since and i'm already seeing signs of growth. :rockon:


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Re: hairy mycelium? [Re: flipsidetrue]
    #19062315 - 10/31/13 09:06 AM (10 years, 2 months ago)

:thumbup:


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Re: hairy mycelium? [Re: PirateSwazey]
    #19066612 - 10/31/13 10:18 PM (10 years, 2 months ago)

so the fresh tissue i threw on some agar hoping for some decent mycelium is looking like cobweb mold also dammit!


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Edited by flipsidetrue (10/31/13 10:38 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19066683 - 10/31/13 10:26 PM (10 years, 2 months ago)

are you cloning a fruit and putting it on agar?
if so, was there any cobweb that you noticed present on your substrate at any time?

if I were you I would clean and lysol-bomb the hell out of the room you're working in, wash out your still air box (if you're using that and not a flowhood) with hot soapy water, and either PC your scalpel to sterilize it or replace your scalpel altogether.  (that happened to me, i had a few dishes in a row get mold and my dishes have historically always been clean - i replaced the scalpel i was using with a new one and the mold hasn't returned)  so just throwing that out there for ideas to try


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Re: hairy mycelium? [Re: PsiLisaBin]
    #19066694 - 10/31/13 10:29 PM (10 years, 2 months ago)



I am using a still air box and not a flow hood, and i am trying to clone from a fruit that i grew outside because the cake i was doing had green mold on it. 4 months ago i got some spores from a vendor i was doing the pf tek and i got some fruits off of it, then i made some spore sprints and kept making syringes, and kept using them to knock up some more until i moved to grains, then when i moved to grains all the cakes i kept doing all got contaminated so i threw em outside still got a decent flush, but i made spore prints from those also and kept knocking up wbs jars, thinking everything was fine but the cakes just kept getting contamed  and i kept throwing them outside until it got to a point to where i only got a few fruits from it so i used those few fruits i got to try and clone and here's where im at now im sure there had to be plenty of cobweb mold associated with those and plenty of other contams


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Edited by flipsidetrue (10/31/13 10:36 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19066785 - 10/31/13 10:41 PM (10 years, 2 months ago)

Man it must be in the air or on your samples.

Cobweb is rare and I suppose you are dealing with the same cobweb spores from before.

You could spray your sample with some h202


Edited by MUSH HEAD420 (10/31/13 10:42 PM)


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Re: hairy mycelium? [Re: MUSH HEAD420]
    #19066876 - 10/31/13 10:56 PM (10 years, 2 months ago)

ah, ok.

i do know it's hard to get a clean print off a mushroom from the great outdoors.  am assuming it is also hard to get a piece of clean tissue from a mushroom grown outdoors as well :wink:

i know you're taking it from inside the stem, so it should be relatively more protected, but the whole fruit itself is going to be "dirtier" since it's been outside versus one from inside your indoor fc.  from the looks of it, you may just not be able to get a clean sample from your outdoor cake fruits.  if you could get even a little mushroom myc going on your dish, you could transfer out away from the contam, but cobweb takes over so fast this may well be impossible to accomplish here.  i'd love to see someone more experienced with outdoor growing chime in here - if it were me, i'd probably scrap this particular endeavor and start again with an entirely new thing (clean spore syringe to agar).


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Re: hairy mycelium? [Re: PsiLisaBin]
    #19066933 - 10/31/13 11:05 PM (10 years, 2 months ago)

He would use a spore syringe if he wanted, I think he wants to try and isolate characteristics from the wild specimen.

The best way imo would be to use h2o2 to combat the cobweb mold, then transfer at least once.

There will be a chance of cobweb ending up on your cakes or grains though.


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Re: hairy mycelium? [Re: MUSH HEAD420]
    #19067005 - 10/31/13 11:20 PM (10 years, 2 months ago)

at first i did use a spore syringe on some agar and i had the same problem. I took some advice from someone that said instead of using a spore syringe to just use actual tissue and clone



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Re: hairy mycelium? [Re: flipsidetrue]
    #19067012 - 10/31/13 11:21 PM (10 years, 2 months ago)

where the fudge is it coming from?!?


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Re: hairy mycelium? [Re: flipsidetrue]
    #19067016 - 10/31/13 11:22 PM (10 years, 2 months ago)

i really didn't want to order more spores because to do so consists of me having to get a p.o box in another state and picking them up from there :mad2:


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Re: hairy mycelium? [Re: MUSH HEAD420]
    #19067026 - 10/31/13 11:24 PM (10 years, 2 months ago)

That's what I'm saying! I'm over here growing grey hairs about to look like Albert Einstein over here


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Re: hairy mycelium? [Re: flipsidetrue]
    #19067040 - 10/31/13 11:27 PM (10 years, 2 months ago)

Well if it happened with the spores and the sample so it did not come from the syringe or the sample.. unless one introduced it to the other.

I am guessing it is in the environment, try and do it outside or something and then seal your agar plates with mircopore tape.


Edited by MUSH HEAD420 (10/31/13 11:29 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19067045 - 10/31/13 11:29 PM (10 years, 2 months ago)

Do u have more fruits/tissue samples? Did you take prints?


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Re: hairy mycelium? [Re: cronicr]
    #19067054 - 10/31/13 11:30 PM (10 years, 2 months ago)

yes i have a couple more prints and I'm down to 1 more fruit and I'm not to confident about the prints being clean


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Edited by flipsidetrue (10/31/13 11:31 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19067140 - 10/31/13 11:50 PM (10 years, 2 months ago)

Sometimes prints can be easier to clean with wild fruits  but with tissue i use to use a method rr wrote with a series of rinses in sterile water in half pints where u just sterilize a few jars of water drop your fruit in and shake, u can add laundry bleach to it if you want but I never did, if I still had problems I turn to cardboard to get some growth and clean that on agar, just my two cents but just be more persistent then the contam lol


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Re: hairy mycelium? [Re: cronicr]
    #19067230 - 11/01/13 12:07 AM (10 years, 2 months ago)

hmmmmm interesting... i think i might try to mix up some h202 solution to help fight the contaminants and then keep transferring until i get a solid healthy growth.. what kind of h202 mixture would you recommend?


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Re: hairy mycelium? [Re: flipsidetrue]
    #19067272 - 11/01/13 12:14 AM (10 years, 2 months ago)

1:10 - 2:10 of 3% peroxide, you can use 1/2 strength 3% peroxide at first to kill the stuff. do not over or under do it.


Edited by MUSH HEAD420 (11/01/13 12:17 AM)


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Re: hairy mycelium? [Re: MUSH HEAD420]
    #19068848 - 11/01/13 08:45 AM (10 years, 2 months ago)

I've been told over and over by the likes of RR and a few trusted cultivators to not shock the mycellium with H2O2 when using clone tissue to agar. You want the healthy myc to out run the contams. Cobweb is tricky but if you're seeing it with spores and your clone tissue I have a feeling it's your sterile technique. It's probably getting in there from some part of pouring your plates or when you open them to put the clone on. Double check your SAB/sterile techniques.

If you want to use the clone tissue from that outdoor fruit. Rip the fruit in half inside your SAB from the base. You should have virgin never seen air tissue on the inside of the stem that you can carefully transfer to the agar. You want to be extra careful not to let any part of your tool touch the outside surface of the fruit just get inside the stem tissue.

Unfortunately I dont think h2o2 is going to help you. It never helped me out and actually made the cobweb run the dishes even more vigorously since it's pretty hard to catch every last drop of cobweb with an h2o2 dip.

You're going to want to make as many dishes as you can from the clone tissue or spore print to start. That way if you make 6 and only 1 is success you can transfer that one to 6 more and toss the 5 that sucked. When starting from such dirty original samples you're going to want probability and numbers on your side so do 10 dishes if you can.

Use parafilm or saran wrap to wrap the dishes.


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Re: hairy mycelium? [Re: bodhisatta]
    #19068864 - 11/01/13 08:53 AM (10 years, 2 months ago)

Have you tried this:

Go through the whole agar process as normal, except don't actually inoculate.

Do the open and close the dish, parafilm, the whole nine. Let the dish sit and see if it contaminates.


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Edited by SpitballJedi (11/01/13 08:55 AM)


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Re: hairy mycelium? [Re: SpitballJedi]
    #19069448 - 11/01/13 11:33 AM (10 years, 2 months ago)

@spitballjedi: I've done something very similar to that actually, when i first got my MEA i went ahead and poured into several dishes so all i would have to do was inoculate or place tissue into, and the ones that i poured and left alone actually had no contaminants

this is the SAB i'm using



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Edited by flipsidetrue (11/01/13 12:11 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19069488 - 11/01/13 11:43 AM (10 years, 2 months ago)

Btw let me add i live in a household with 4 dogs 2 cats that are constantly running around :crazy:


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Re: hairy mycelium? [Re: flipsidetrue]
    #19069841 - 11/01/13 01:05 PM (10 years, 2 months ago)

Quote:

flipsidetrue said:
i really didn't want to order more spores because to do so consists of me having to get a p.o box in another state and picking them up from there :mad2:





that does suck


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Re: hairy mycelium? [Re: flipsidetrue]
    #19069937 - 11/01/13 01:31 PM (10 years, 2 months ago)

Just for the sake of being thorough, When you poured your plates before and just let them sit, were they wrapped in the same tape as the above pictures? Were they stored/kept exactly the same way as your inoculated dishes.

I'm just trying to help see if there is some subtle detail that's different.

In my experience, most people don't have the exact same contams when using spores AND tissue unless there is something wrong about the technique. Sometimes it's very subtle.


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Re: hairy mycelium? [Re: SpitballJedi]
    #19069991 - 11/01/13 01:44 PM (10 years, 2 months ago)

the plates that i did pour and let sit i didn't wrap them in any tape or Saran wrap because i read somewhere that it wasn't necessary if  tissue/ spores weren't added to them, i actually just sat them in my closet and they had no signs of contams until about 48 hours after i added the tissue/ spores


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Edited by flipsidetrue (11/01/13 01:46 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19070177 - 11/01/13 02:23 PM (10 years, 2 months ago)

Did you leave the poured/blank ones in the original bag or something?


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Edited by SpitballJedi (11/01/13 02:25 PM)


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Re: hairy mycelium? [Re: SpitballJedi]
    #19070279 - 11/01/13 02:50 PM (10 years, 2 months ago)

nope i didnt even do that much i just sat them on a shelf in my closet


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Re: hairy mycelium? [Re: flipsidetrue]
    #19070358 - 11/01/13 03:10 PM (10 years, 2 months ago)

:utterconfusion:


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Re: hairy mycelium? [Re: SpitballJedi]
    #19070483 - 11/01/13 03:36 PM (10 years, 2 months ago)

i know right? so i think what I'm going to do is spray the inside with some bleach/water solution,then go to town with my brand new x ac-to knife after flame sterilization of course. But before i do anything i have a concern about something which is i had a brand new sleeve of Pre-sterilized petri dishes that i opened inside the SAB but i didnt use all 20 of them, I'm thinking i might have messed up by doing so? :eek:


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Edited by flipsidetrue (11/01/13 05:42 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19072546 - 11/01/13 10:15 PM (10 years, 2 months ago)

use h2o2 not bleach.. imo


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Re: hairy mycelium? [Re: MUSH HEAD420]
    #19074527 - 11/02/13 10:41 AM (10 years, 2 months ago)

alright note taken i wanna thank everyone that chimed in on this post for your advice and help! MUCH LOVE!:dawerp:


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Re: hairy mycelium? [Re: flipsidetrue]
    #19095197 - 11/06/13 12:41 PM (10 years, 2 months ago)

Alright so this is my 2nd attempt and this time i was more sterile with the procedure.

The first 3 are from a spore print and the last 2 are tissue.





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Re: hairy mycelium? [Re: flipsidetrue]
    #19095224 - 11/06/13 12:45 PM (10 years, 2 months ago)

ya idk man, this looks pretty bad. 

maybe the last one is good :shrug:


Edited by MUSH HEAD420 (11/06/13 12:46 PM)


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Re: hairy mycelium? [Re: MUSH HEAD420]
    #19095230 - 11/06/13 12:47 PM (10 years, 2 months ago)

yea I'm not having the best luck with my agar adventures, i thought the last one looked the best because it looks like transparent mycelium so far with no fuzzy hairs, i think i'll try and transfer part of that one tonight


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Edited by flipsidetrue (11/06/13 01:32 PM)


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Re: hairy mycelium? [Re: flipsidetrue]
    #19095596 - 11/06/13 02:13 PM (10 years, 2 months ago)

The first three could be pin mold.
I thing the last one is just bacteria on the agar. Does the piece of the mushroom have myc growing on it or is that just glare from the camera?


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Re: hairy mycelium? [Re: Stupendous-Yappi]
    #19096371 - 11/06/13 04:54 PM (10 years, 2 months ago)

first 3 are cobweb again. bottom left looks lke bacteria, but the one on the right looks like weak myc growth. see how it looks after another day or so.


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Re: hairy mycelium? [Re: Stupendous-Yappi]
    #19096540 - 11/06/13 05:24 PM (10 years, 2 months ago)

the bottom left it looks like some milky shit its bacteria probably and the bottom left one thats not a glare it has stuff growing from the tissue i was thinking its cobweb because of how hairy/stringy and how fast its growing


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Re: hairy mycelium? [Re: PirateSwazey]
    #19096550 - 11/06/13 05:25 PM (10 years, 2 months ago)

sweet i'll wait a day or so and post some pic of what seems to be weak mycelium growth, this is the break i've been hoping for!


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