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Offlinefrankwhite85
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Registered: 04/18/16
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vectors for bacteria
    #23933791 - 12/16/16 09:48 PM (7 years, 1 month ago)

Have been experimenting with putting ms agar wedges to grain but the jar either stalls after a shake or fully colonizes but smells off when I go to do a g2g.  I pc my rye jars for at least 90 min and all innoculations are done in front of a flowhood.  My jars dont have outright contamination like spots of green or mushey ice cream looking mycelium they look great it just always has that off smell.  My cultures that I inoculate with look great too, never any visible mold or bacteria spots So I'm not sure what's going wrong.  Is my sterile technique/hand movements in front of the hood to blame or is it something else?


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InvisiblePirateSwazey
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Re: vectors for bacteria [Re: frankwhite85]
    #23933798 - 12/16/16 09:52 PM (7 years, 1 month ago)

Take some pics.

Sounds like paranoia to me...

Are you sure they're stalling?

How much growth before you shake?

Colonization temps?


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OfflinePsilosoulful

Registered: 09/05/14
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Re: vectors for bacteria [Re: PirateSwazey]
    #23933802 - 12/16/16 09:54 PM (7 years, 1 month ago)

Take enough clean transfers from plates to others before deciding to drop it in a grain jar


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InvisiblemushboyMDiscord
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Re: vectors for bacteria [Re: Psilosoulful]
    #23933806 - 12/16/16 09:56 PM (7 years, 1 month ago)

ive had a few cultures that would not 'jump' onto the grain. but only a few out
of a whole lot. how many are you doing?


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OfflinePsilosoulful

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Re: vectors for bacteria [Re: mushboy]
    #23933820 - 12/16/16 09:59 PM (7 years, 1 month ago)

Same, the wedges seem to dry out completely before getting a chance to jump to the grains.
Then again I was using oats, which never worked for me anyways, the moisture content was always off


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InvisiblemushboyMDiscord
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Re: vectors for bacteria [Re: Psilosoulful]
    #23933839 - 12/16/16 10:09 PM (7 years, 1 month ago)

OP should try munches blenderless LI.

i havent yet but it might help you out.


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Offlinefrankwhite85
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Re: vectors for bacteria [Re: PirateSwazey]
    #23933924 - 12/16/16 11:02 PM (7 years, 1 month ago)

Here are some pics of a B+ culture from a few transfers back.  This B+ culture is the only one that looks decent (I tried a GT one that didn't work out so well).  Each time I take a round of transfers from the best looking cultures I put the remaining culture I took a transfer from to grain too.  On the cultures I've put to grain they would look like this after about a week from inoculation in a room set at 75 degress. I would let it colonize probably 2 layers of grain deep surrounding the wedge then give it a shake wait probably another week for it to get about 30% colonized then give it another shake. When I've been doing that most of the jars just don't even look close to colonizing within a month and the ones that do usually take 3-3 and 1/2 weeks and end up smelling funky.  I actually just tried blenderless LI a few days ago I'm excited to see how much faster colonization times can be.











Edited by frankwhite85 (12/16/16 11:06 PM)


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OfflineDactylium
Don't touch me I'm sterile


Registered: 08/12/16
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Re: vectors for bacteria [Re: frankwhite85]
    #23933932 - 12/16/16 11:11 PM (7 years, 1 month ago)

Maybe try bumping your PC time up to 2 hours and continue with the blenderless LI. It could be that the endospores are bouncing back while your lack of inoculation points takes ages to colonize. Maybe shine a light through your cultures to see if there's anything hiding in there. Have you been wrapping your dishes in plastic film? Something could have come through the vents.  :shrug:

Sneaky contaminants


Edited by Dactylium (12/16/16 11:17 PM)


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InvisiblePirateSwazey
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Re: vectors for bacteria [Re: Dactylium]
    #23933949 - 12/16/16 11:19 PM (7 years, 1 month ago)

Your cultures look good man.

You need to shuffle the grains after you drop your wedges.

They will colonize a lot better if the wedges are spread out throughout the jar.

Maybe you're shaking too early.

I shake once at about 20% colonization.

Fully colonized masters in about 3 weeks from inoculation.

Are you working with clones or MS?


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Edited by PirateSwazey (12/16/16 11:46 PM)


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Offlinefrankwhite85
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Re: vectors for bacteria [Re: PirateSwazey]
    #23934060 - 12/17/16 12:03 AM (7 years, 1 month ago)

MS I'm hoping to get some tubs going to clone some fruits soon.  I really think getting LI down will help speed up colonization times I just think it's taking so long that it my window of sterility is running out so bacteria is starting to rear its ugly head. and I use parafilm every time.


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OfflineDactylium
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Re: vectors for bacteria [Re: frankwhite85]
    #23934087 - 12/17/16 12:14 AM (7 years, 1 month ago)

Give that a go and I don't see how you could fail. Your technique is on point judging by the looks of those cultures. I was just throwing everything out there when I suggested looking for hidden contams.


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InvisiblePirateSwazey
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Re: vectors for bacteria [Re: frankwhite85]
    #23934103 - 12/17/16 12:18 AM (7 years, 1 month ago)

Liquid was never any faster for me... I just see it as an unnecessary vector for contamination. Wouldn't recommend myself :shrug:


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Offlinefrankwhite85
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Re: vectors for bacteria [Re: PirateSwazey]
    #23934171 - 12/17/16 12:49 AM (7 years, 1 month ago)

Did you do liquid inoculant or liquid culture? I think if its done right it would go way faster, that's going from a few inoculation points to a couple hundred.


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OfflineDactylium
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Re: vectors for bacteria [Re: frankwhite85]
    #23934191 - 12/17/16 12:57 AM (7 years, 1 month ago)

Well what Pirate is suggesting is shaking the wedge around in the grain which would break off microscopic pieces of mycelium, giving you a bunch of inoculation points. That could probably work just as well. I don't know that anyone has ever done an A/B with an isolate to see if LI is any faster. I'd recommend shaking the wedge and seeing if that works first. If it does I don't see any reason to mess with LI if you don't have to.


Edited by Dactylium (12/17/16 01:00 AM)


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InvisibleAdden
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Re: vectors for bacteria [Re: frankwhite85]
    #23934194 - 12/17/16 12:59 AM (7 years, 1 month ago)

Use clean LI, better sterile tek, maybe check your grain prep.

What happens to a jar when you just leave it laying around?


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Offlinefrankwhite85
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Re: vectors for bacteria [Re: Adden]
    #23934201 - 12/17/16 01:06 AM (7 years, 1 month ago)

Like Pc'd but uninnoculated? haven't tried it but I'd assume nothing I cant remember the last time I had outright visible contamination its always invisible bacteria which I'm guessing would only come from dirty cultures or bad sterile technique in front of flowhood.


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InvisiblePirateSwazey
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Re: vectors for bacteria [Re: frankwhite85]
    #23934211 - 12/17/16 01:10 AM (7 years, 1 month ago)

Quote:

frankwhite85 said:
Did you do liquid inoculant or liquid culture? I think if its done right it would go way faster, that's going from a few inoculation points to a couple hundred.



I have used both LI and LC plenty. Nothing has been as successful as plain old agar to grain though. Like Dactylium said shaking the wedges knocks off microscopic pieces of mycelium and it gets spread all around the jar.


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Invisiblevan hattonFacebookDiscord
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Re: vectors for bacteria [Re: PirateSwazey]
    #23934302 - 12/17/16 02:12 AM (7 years, 1 month ago)

Quote:

I would let it colonize probably 2 layers of grain deep surrounding the wedge then give it a shake wait probably another week for it to get about 30% colonized then give it another shake.




Just shake it when you drop along with what everyone else is saying.

My a2g jars generally take 10-14 days to be 100℅ my g2gs now take like 8 days but I only do it too 7 at a time.

I've been loving the quick colonization times. Less time for anything else to take hold.

:cheers:
Van


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Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

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OfflineNDStepp84
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Re: vectors for bacteria [Re: van hatton]
    #23934505 - 12/17/16 04:49 AM (7 years, 1 month ago)

Upping your PC time as mentioned couldn't hurt, the minimum should be 2 hours IMO and check your filters.


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:takingnotes: Links and teks:takingnotes:
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Offlinewtfcrazymofo
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Re: vectors for bacteria [Re: NDStepp84]
    #23934549 - 12/17/16 05:44 AM (7 years, 1 month ago)

Shake your dropped wedges almost as hard as a blenderless innoc tek, and Use bigger wedges.  Get that wedge to touch every grain in the jar, and It won't take much longer to get to 100% than A LI.


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If you want to eat->https://www.shroomery.org/forums/showflat.php/Number/8553541

Bag sealers are to bulky (my hood isn't that big)
https://www.shroomery.org/forums/showflat.php/Number/28622922




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Offlineglowdiver
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Re: vectors for bacteria [Re: wtfcrazymofo]
    #23934816 - 12/17/16 08:55 AM (7 years, 1 month ago)

I'm still pretty much a noob, but I wonder about your lids.  Could the stalling be due to lack of ge? None of your pics show your lids.  What are you doing for gas exchange, 1/4" holes with filter discs, tyvek, loosening, etc.?  Or maybe I'm way off base.


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