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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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slant issues
#24879789 - 12/30/17 09:49 PM (7 years, 18 days ago) |
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I have been collecting cultures for about a year and a half now and i have amassed a pretty decent collection at this point ( probably 20 species and several different strains of some). i decided i better start making slants so i dont lose them. I bought a huge bag of 15ml pp5 centrifuge tubes to use. for some reason around 15% of them dont seem to grow at all, the transfer will get fuzzy like it should , but it never seems to expand onto the agar, i am using the same recipe as i use for my dishes. 2% MEA im using good agar from fungi perfecti. the dishes have great growth. i cannot see any kind of bacterial growth on the surface or any off color, they look exactly the same as the ones that do grow. i made multiples of each culture and in most cases it was just 1 of three that didnt grow, so its not the cultures either.im not screwing the caps down tight i back them off a bit and wrap with parafilm. has anyone else experienced anything like this before? i just did a surface scrape of one that hadn't moved for 2 weeks now and rubbed the loop on a clean agar plate to see if maybe i am just missing a very light colored bacterial or yeast infection but i really dont think that is my problem. i just picked up 30 big 25x200 glass test tubes i am going to make some more masters on of all my really good cultures but i was hoping for some insight into this before i made anymore slants and waste more time and materials. thanks in advance for your thoughts and ideas.
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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So you have good growth on 2% MEA on petri dishes but then you don´t have any growth or recovery on 2% MEA in slant tubes? That sound really weird.. Did I understand correctly?
-------------------- Cakes inside Water Tub
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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Re: slant issues [Re: Mateja]
#24879922 - 12/30/17 10:00 PM (7 years, 18 days ago) |
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that is correct
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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That makes no sense at all. Can´t be GE issues, can't be the agar media. Can you post some pics of the problematic tubes?
-------------------- Cakes inside Water Tub
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
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Re: slant issues [Re: Mateja]
#24879938 - 12/30/17 10:05 PM (7 years, 18 days ago) |
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sure, let me see if i can get this to work on my phone, it will probly take a few mins at least.
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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Shu
Vote for Humanity



Registered: 11/20/11
Posts: 885
Loc: PA, USA
Last seen: 6 days, 19 hours
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Quote:
mattthemushroomman said: the transfer will get fuzzy like it should , but it never seems to expand onto the agar,
Most likely insufficient gas exchange imo.
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
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First pick is pe done 11/17 no growth. Second is H. Americanum from 12/17 and third is also H.a done same day same everything
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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hamloaf
Q-dood ®©™√



Registered: 12/23/09
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The culture isn't going to last/store for nearly as long with out the popsicle stick.
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
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Here is what it looks like on a plate, def growing well
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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Yeah, I realize now I should have a coffee sitter in there, I will with my next batch. Glad I bought 100 lol
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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How did you mix your agar for the slants? I use premix and had troubles in the past where I didn't get a good mixture.
For my slants I measure normally and mix in a measuring cup, stirring with a spoon like a cup of ovaltine between filling up tubes.
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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Re: slant issues [Re: Shu]
#24880055 - 12/30/17 10:28 PM (7 years, 18 days ago) |
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i leave the caps partially unscrewed so they can get GE, here is a thought on that note though, i did these no pour, i put 7 ml in each then filled jars with them pced and then layed in front of the hood to cool. does the pc cook all the O2 in the slant? maybe i just need to leave them all open fora bit in the flow of the hood?
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
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Last seen: 4 years, 6 months
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1 l of water, 20g malt extract and 20g of agar in a pot on the stove, once all well mixed i use a baster to fill each slant
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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Quote:
mattthemushroomman said: Here is what it looks like on a plate, def growing well
What culture is this?
And no the PC does not "cook" the O2, even if it did, you´d still have to let in new air when you inoculate the slants. Anyway that´s surely not the problem. GE is not a problem either so early in the transfer. This is really weird. And it doesn't seem to be unevenly mixed agar media either so this is a mystery to me
-------------------- Cakes inside Water Tub
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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Re: slant issues [Re: Mateja]
#24880070 - 12/30/17 10:36 PM (7 years, 18 days ago) |
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The plate is Hericium americanum i believe. it could possibly be coralidies. i have some growing out right now but i dont have my air exchanges dialed in right yet and i dont have a co2 meter to see how bad it is in there. i need to get a good waterproof fan i will post a pic of the fruits in a few mins
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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Re: slant issues [Re: Mateja]
#24880077 - 12/30/17 10:41 PM (7 years, 18 days ago) |
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Here is the fruits
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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Quote:
mattthemushroomman said: The plate is Hericium americanum i believe.
On the dish the culture didn't seem to grow unilaterally at all so it´s hard to imagine it would fill out the surface of the media in the tub completely. If it was cube mycelium I would give you better answer but I don´t have experience with Hericium..
-------------------- Cakes inside Water Tub
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mattthemushroomman
mycoculturist


Registered: 04/27/16
Posts: 56
Loc: USA
Last seen: 4 years, 6 months
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Re: slant issues [Re: Mateja]
#24880092 - 12/30/17 10:53 PM (7 years, 18 days ago) |
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if you lo at the second of those three pics of tubes you can see the hericium has spread across the surface pretty decent, the thrid pic is the same culture but it hasnt moved. the first pic is a cube but its PE and that one is a month and a half old now. Thank you either way for your thoughts.
-------------------- working on a business growing gourmet mushrooms and supplies for growing.
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Shu
Vote for Humanity



Registered: 11/20/11
Posts: 885
Loc: PA, USA
Last seen: 6 days, 19 hours
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I cannot see the type of cap you have but often they have a conical compression lip inside (vs a flat cap with a gasket) that even though you may think it's loose may still be making contact with the tube. I suggest simply giving them another quarter turn open.
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mushboy
modboy



Registered: 04/24/05
Posts: 35,378
Loc: eating the cats
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Re: slant issues [Re: Shu]
#24880550 - 12/31/17 08:09 AM (7 years, 17 days ago) |
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Are slants media supposed to be less nutritious or more or doesnt matter?
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hamloaf
Q-dood ®©™√



Registered: 12/23/09
Posts: 24,389
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Re: slant issues [Re: mushboy]
#24880556 - 12/31/17 08:11 AM (7 years, 17 days ago) |
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Slant media is the same.
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mushboy
modboy



Registered: 04/24/05
Posts: 35,378
Loc: eating the cats
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Re: slant issues [Re: hamloaf]
#24880580 - 12/31/17 08:27 AM (7 years, 17 days ago) |
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Thanks brotradamus
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