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Amazon Shop for: ½ Pint Jars, Gypsum, Laminar Flow Hood, Wild Bird Seed

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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 11 years, 8 months
black dot contam in 100% colonized WBS?
    #2008633 - 10/14/03 05:05 PM (13 years, 1 month ago)

been getting this black dot contam recently in the WBS. G2G seemed to eliminate the problem then the two jars that were furthest along both have a tiny little start of contam towards the bottom. Everything on top is 100% pretty white. Is this top layer of the quart jar okay or should they be tossed and wait for a few more better jars to do more transfers?


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InvisibleShamanorrhea
Stranger
Registered: 06/20/03
Posts: 304
Re: black dot contam in 100% colonized WBS? [Re: thisone]
    #2008813 - 10/14/03 05:59 PM (13 years, 1 month ago)

If you're doing grain to grain transfers and not using peroxide then you are risking contamination. I'll include a few links for you to use at your disposal.

http://mv.lycaeum.org/peroxide
http://mv.lycaeum.org/peroxide/_peroxide_summary.html
http://www.mycomasters.com/Try-it.html
http://www.wildmushrooms.org/frames/oms_peroxide.htm
http://members.aol.com/PeroxyMan/


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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 11 years, 8 months
Re: black dot contam in 100% colonized WBS? [Re: Shamanorrhea]
    #2008922 - 10/14/03 06:28 PM (13 years, 1 month ago)

Sweet! Thanks for including those. so does H202 really help that much? So you inject 12 mL (is that 12cc's) after pressure cooking after it's cooled?


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OfflineDMJ
creepyatmosphere

Registered: 10/10/03
Posts: 233
Loc: Dungeon
Last seen: 7 years, 4 months
Re: black dot contam in 100% colonized WBS? [Re: thisone]
    #2009000 - 10/14/03 07:00 PM (13 years, 1 month ago)

I'm quite sure that Milliliters is less than cubic centimeters.


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Offlinejumpship
lawn boy

Registered: 08/16/03
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Re: black dot contam in 100% colonized WBS? [Re: DMJ]
    #2009069 - 10/14/03 07:35 PM (13 years, 1 month ago)

negative, a ml is equal to a cc.


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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
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Re: black dot contam in 100% colonized WBS? [Re: jumpship]
    #2009131 - 10/14/03 07:53 PM (13 years, 1 month ago)

so one whole syringe of h202? Will that throw off the water ratio?


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Offlinestoner2002
Stoner

Registered: 11/15/02
Posts: 183
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Re: black dot contam in 100% colonized WBS? [Re: thisone]
    #2009372 - 10/14/03 09:25 PM (13 years, 1 month ago)

that cant be right, 12 ccs?! thats a lot of excess water


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Invisiblemicro
bunbun has a gungun
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Re: black dot contam in 100% colonized WBS? [Re: stoner2002]
    #2009430 - 10/14/03 09:44 PM (13 years, 1 month ago)

H202 is used at 5-20cc's (ml's) per liter water or media or whatever when *taken out of a 3% solution.* This is the concentration in almost all H2O2 you'll buy in a store. Find details at www.mycomasters.com.

You must add the H2O2 when the solution into which you're putting it is cool (or only a little warm) to the touch.

If you see black dots growing from the mycelium it's probably spores -- P. cubensis mycelium doesn't sporulate, so I would think there's something wrong -- as long as you've seen it growing.

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Offlinestoner2002
Stoner

Registered: 11/15/02
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Re: black dot contam in 100% colonized WBS? [Re: micro]
    #2009684 - 10/14/03 11:04 PM (13 years, 1 month ago)

Does anyone do grain to grain transfers WITHOUT peroxide?


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Invisiblemicro
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Re: black dot contam in 100% colonized WBS? [Re: stoner2002]
    #2009695 - 10/14/03 11:08 PM (13 years, 1 month ago)

Yes, but in a laminar flow hood.

Instant ~ 500 cc's inoculum:

Take 500 cc's water and add ~ 50 cc's 1mm glass beads. Sterilize. Sterilly throw in several chunks of mycelium, close and shake. Draw up with sterile syringes in a laminar flow hood.

--
Micro


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Offlinethisone
the one theydon't see

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This sucks big time. Half grain jars contam at 100% [Re: micro]
    #2011216 - 10/15/03 12:54 PM (13 years, 1 month ago)

Those f*cking black dot contams got in there again, those little bastards. bleached the jars, tried experimenting with extra pc time aroun 90 min to 120 min at 16 psi to make sure everything was history. Was all super exited about this grain to grain thing because everything took off so quickly and now once they reached 100% they have the blak dot too around the mycelium so I know it's contams not spores. What's up with the H202, someone has to know the answer? What about gypsum? Would that serve the same purpose without having to worry about the excess moisture issue? Please, Please help. I'm ready to say fuck grain, thrown out over 60 pounds of birdseed because of contams. Want to stay vigilant in this battle.


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Invisiblemicro
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Re: This sucks big time. Half grain jars contam at 100% [Re: thisone]
    #2011245 - 10/15/03 01:15 PM (13 years, 1 month ago)

I posted the concentration of H2O2 to use above; read it. I probably didn't explain the spore thing well -- it's likely a contaminant that's sporulating -- maybe Rhizopus. It has a white mycelium, and can look like cubensis mycelium, but the black dots form when it sporulates. If the black dots THEMSELVES are growing, though, it's some contaminant.

I guess you just have to be more sterile. Are you doing this in a glovebox? If you tell me the setup and how you do things I could comment....

--
Micro


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(Avatar is Maxxy, a character by Mizzyam, RIP)


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Offlinethisone
the one theydon't see

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Posts: 818
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Re: This sucks big time. Half grain jars contam at 100% [Re: micro]
    #2011254 - 10/15/03 01:30 PM (13 years, 1 month ago)

first time tried a glovebox, black dot contam. Second time sterile bathroom, black dot contam. Another time spore syringe, black dot contam. Another time G2G, black dot contam. Yes I am 100% sure it is contmaniation. I understand what you mean about sporulating. This is contamination for sure. What's weird is that with the G2G it only occured at the 100% colonization level and was fine the whole time. Even now it is just starting to contam, not spread all over like crazy. I don't understand what you wrote about adding the H202. Could you explain that more. They way I heard was to inject it in the grain before transfering colonized WBS. then shake it up so the colonized WBS is distribute evenly along with the H202. Is this right? What about the gypsum. There has to be more people who do grain. Anyone else struggled with it like this. I don't know maybe grain is not for me. I am being very, very, very sterile so I don't know what's going on.


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Offlinethisone
the one theydon't see

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Posts: 818
Last seen: 11 years, 8 months
Re: This sucks big time. Half grain jars contam at 100% [Re: thisone]
    #2011261 - 10/15/03 02:37 PM (13 years, 1 month ago)

been reading a bit on steeping and soaking. Never did the steep just the soak. Is the steep supposed to be first? Also read a lot a people have sucess with the birdseed pcing at 2 hours+?


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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 11 years, 8 months
Re: This sucks big time. Half grain jars contam at 100% [Re: thisone]
    #2011993 - 10/15/03 06:43 PM (13 years, 1 month ago)

please Help?


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Amazon Shop for: ½ Pint Jars, Gypsum, Laminar Flow Hood, Wild Bird Seed

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