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Offline6Curi0us.M0nkey9
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Lion’s mane: Blue mycelium on agar
    #26861285 - 08/03/20 05:33 PM (3 years, 5 months ago)



Hello, I am new to posting as well as cultivating; so I apologize in advance if the images were uploaded incorrectly. Shown here are 3 agar dishes that are being colonized by Lion’s Mane (and possibly an unwelcome mold?).

The first one is the parent dish that I inoculated with a liquid culture. I have cut a good deal of contaminants out, as well as cut some out for transfer to the other two dishes. The other two dishes are a result of the transfers. The agar was made with barley malt extract and nutritional yeast.

Question 1: Why does the mycelium on the two transfer dishes have a blue tint?

Question 2: If this is contamination, how do I salvage the growth? It doesn’t look very localized.

Question 3: Why is the mycelium growing so irregularly on the transfer dishes? Is it due to an uneven dispersion of nutrients? How can I prevent this?

Question 4:Why is the thick, blue, uneven growth lacking on the parent dish? It looks evenly dispersed and white.

Question 5: Does the parent dish look dry?

Thanks in advance for any help provided. Please ask questions if additional information is needed to properly advise me.


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“And those who were seen dancing were thought to be insane by those who could not hear the music”
                -Friedrich Nietzsche


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OfflinePhrontist
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9] * 1
    #26861293 - 08/03/20 05:37 PM (3 years, 5 months ago)

Quote:

6Curi0us.M0nkey9 said:





Unless every one of those spots is a place where you transferred agar, I’d say you 100% have bacterial contamination.


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OfflineSHROOMSISAY01M
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Re: Lion’s mane: Blue mycelium on agar [Re: Phrontist]
    #26861299 - 08/03/20 05:41 PM (3 years, 5 months ago)

Start over all of those spots are mold.


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: SHROOMSISAY01]
    #26861345 - 08/03/20 05:59 PM (3 years, 5 months ago)

Thank you for the timely reply, one less thing to learn for me! Thanks a lot.


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Offlinesendmehummus
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26861460 - 08/03/20 07:15 PM (3 years, 5 months ago)

You have a wonderful collection of mold mold and more mold! Beginner tip, every mycelium is pure white. Makes it easy to spot any kind of contamination. My advice? (And this should answer all your questions).. start completely over. Maybe you can somehow pull off the impossible and get a pure dish after a dozen transfers but youre going to waste time, agar, dishes and money. Just get a LM liquid culture syringe, inoculate a ton of grain jars and use the last few drops on a dish or two. Thats how I start my dishes. Usually pure mycelium from the get go, if not Ive had to transfer like twice at the most. Youve just gotta scrap that contamination nation and start over brotha! Good luck! 😊✌️


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Be patient and keep it simple.


Edited by sendmehummus (08/03/20 07:17 PM)


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: sendmehummus]
    #26861642 - 08/03/20 09:01 PM (3 years, 5 months ago)

This is fair advice I appreciate it. But why ditch the parent dish? Is it also contaminated?


Edited by 6Curi0us.M0nkey9 (08/03/20 10:18 PM)


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OfflineSHROOMSISAY01M
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26861692 - 08/03/20 09:36 PM (3 years, 5 months ago)

Don't ditch the parent dish. I would assume it is ok. Sorry I should have been more clear.


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OfflineSHROOMSISAY01M
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Re: Lion’s mane: Blue mycelium on agar [Re: SHROOMSISAY01]
    #26861699 - 08/03/20 09:41 PM (3 years, 5 months ago)

Are you sure there is LM mycelium on that parent dish? I don't see anything on it. The dish should be white.


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OfflineSHROOMSISAY01M
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Re: Lion’s mane: Blue mycelium on agar [Re: SHROOMSISAY01]
    #26861707 - 08/03/20 09:46 PM (3 years, 5 months ago)

Here is my LM Petri dish...



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Re: Lion’s mane: Blue mycelium on agar [Re: SHROOMSISAY01]
    #26861829 - 08/03/20 11:36 PM (3 years, 5 months ago)

I have all white petri dish... but I just cannot transfer it - it doesn't want to grow. Any tips?


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OfflineForresterM
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Re: Lion’s mane: Blue mycelium on agar [Re: Bsdgaou]
    #26862820 - 08/04/20 03:09 PM (3 years, 5 months ago)

OP, are you using a SAB or flow hood? 

That's a LOT of mold spores that got in there :jawdrop:


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26862983 - 08/04/20 04:32 PM (3 years, 5 months ago)

Typical LM, pins generally form (right of center) before full colonization of the plate.



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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: Forrester]
    #26862999 - 08/04/20 04:39 PM (3 years, 5 months ago)

I use a SAB and I thought my sterile technique was sufficient enough to prevent severe contamination; though this is obviously not true.

I always wash my SAB with soap and water, always closing it asap, and keeping closed as much as possible. I wipe everything with isopropyl alcohol before putting it in the SAB, and also give the SAB some sprays of isopropanol. I always heat my syringe red hot. I attempt to keep the dishes closed as much as possible. I always wrap cleanly with parafilm and I store the inoculated dishes on a very high shelf under a small lean-to to prevent excessive airflow. All my poured but “uninoculated”(?) dishes are stored in my refrigerator dedicated to mycology and other sensitive practices (so no food).

Any additional tips or is there anything I’m missing?


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“And those who were seen dancing were thought to be insane by those who could not hear the music”
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863011 - 08/04/20 04:46 PM (3 years, 5 months ago)

Conservation of motion is key within a still air environment. Try to move less with greater efficiency, you may be stirring up turbulence with your arm/hand movements.


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: Stipe-n Cap]
    #26863014 - 08/04/20 04:48 PM (3 years, 5 months ago)

Interesting, thank you so much.


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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863015 - 08/04/20 04:49 PM (3 years, 5 months ago)

Do you wear gloves? Might make a difference. Other than that you may want to try using your SAB in a diff room. If youre still having problems you can clean and place everything youll be working with in the SAB, then seal your arm holes temporarily and let the air settle over a few hours while keeping it in a room with the lowest airflow possible. Cleaning the room that you use your SAB in is as important as cleaning the SAB! Good luck my friend.


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Be patient and keep it simple.


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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863024 - 08/04/20 04:55 PM (3 years, 5 months ago)

I pretty much guarantee that you're going to need to slow down, be more aware of your movements. Try to be aware that every movement will disrupt the stillness of your box, picture it being filled with smoke and how that smoke would swirl and create eddies with each motion, the key is to keep those eddies to an absolute minimum because they're going to carry airborne contaminants. Spray soapy water to trap spores to surfaces, use a dish drying rack to keep your work surface above the contamination on the floor of the box.


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: sendmehummus]
    #26863045 - 08/04/20 05:10 PM (3 years, 5 months ago)

I always wear nitrile gloves in addition to the heavier duty gloves I have secured to the inside of the armholes of my SAB. I always clean the SAB gloves thoroughly. My SAB is completely sealed from its external environment when closed. I also always close the supply air vents in the room. I may move rooms though, I typically do it in my kitchen (most space and best source of water for cleaning).


--------------------
“And those who were seen dancing were thought to be insane by those who could not hear the music”
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InvisibleStipe-n CapMDiscord
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863049 - 08/04/20 05:11 PM (3 years, 5 months ago)

Quote:

6Curi0us.M0nkey9 said:
... in addition to the heavier duty gloves I have secured to the inside of the armholes of my SAB.




Boom, that right there is your issue, get rid of those gloves attached to the arm holes.


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OfflineForresterM
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863050 - 08/04/20 05:12 PM (3 years, 5 months ago)

Agreed.

I never did like that perma-gloves inside the box idea...


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: Stipe-n Cap]
    #26863052 - 08/04/20 05:13 PM (3 years, 5 months ago)

Ahhh, I will certainly have to add the rack to my SAB, that will undoubtedly help. I will minimize my movements. It’s crazy how precise of a practice it is working with agar.


--------------------
“And those who were seen dancing were thought to be insane by those who could not hear the music”
                -Friedrich Nietzsche


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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863055 - 08/04/20 05:13 PM (3 years, 5 months ago)

Definitely get rid of the gloves attached to the SAB. Seems like a good idea in theory but it just leads to more fumbling and more air movement. Also more surface area on the inside of your tub, no good.


--------------------
Be patient and keep it simple.


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: Stipe-n Cap]
    #26863062 - 08/04/20 05:16 PM (3 years, 5 months ago)

Great, I’m glad I now know this. I will remove them asap, they were bulky anyway. You guys are very helpful, thank you.


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“And those who were seen dancing were thought to be insane by those who could not hear the music”
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863066 - 08/04/20 05:18 PM (3 years, 5 months ago)

Literally what we're here for :thumbup:
Good luck on your next round of sterile work.


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Offline6Curi0us.M0nkey9
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Re: Lion’s mane: Blue mycelium on agar [Re: SHROOMSISAY01]
    #26863104 - 08/04/20 05:37 PM (3 years, 5 months ago)

I’m certain of absolutely nothing. It looks like mycelium to me, but this is my first time. It is white, very thread-like, and wispy. It seems to be inside the agar as opposed to on the surface though. I attributed its scattered growth to the fact that is was inoculated with an LC. Therefore the whole dish came into contact with the culture.


--------------------
“And those who were seen dancing were thought to be insane by those who could not hear the music”
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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863115 - 08/04/20 05:41 PM (3 years, 5 months ago)

With LC just put a drop on the plate and streak with an inoculation loop. No need to saturated the entire plate.


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Re: Lion’s mane: Blue mycelium on agar [Re: 6Curi0us.M0nkey9]
    #26863144 - 08/04/20 05:53 PM (3 years, 5 months ago)

Something I did with my SAB which seems to help reduce air flow a little more but while also maintaining easy hand and arm movement is to tape over the front and back of your arm holes with clear gorilla tape. Then cut a cross slit and you have a nice flap that self closes when youre done. I say clear gorilla tape because it has this ideal flexibility that other clear tape (like packaging tape) doesnt have. A cheap and simple thing to do, give it a try :smile: .





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Be patient and keep it simple.


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Re: Lion’s mane: Blue mycelium on agar [Re: sendmehummus]
    #26863152 - 08/04/20 05:58 PM (3 years, 5 months ago)

If you guys plan on using a SAB long term I highly recommend building one, totes are handy but a purpose built SAB is definitely better. You're limited only by your own imagination.  I've always found the tote style SAB design to be tedious.



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Re: Lion’s mane: Blue mycelium on agar [Re: Stipe-n Cap]
    #26863184 - 08/04/20 06:13 PM (3 years, 5 months ago)

Nice! Def seems like a fun project to make a serious SAB. Personally Ive had great success with my tote SAB and Ive strangely come to actually enjoy working with it. Id rather just put any future money towards a cheap flo hood setup (Ive seen some excellent builds for under 700$ on youtube). One advantage the simple tote has for me though is mobility. Ive changed apartments 4x in the last 4 years and taking the tote from place to place (or room to room if need be) is simple as pie. Those pics are cool though 😁👍.


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