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OfflineScarg
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How old is too old for viable spawn?
    #19305167 - 12/20/13 09:06 PM (10 years, 2 months ago)

Posted this at the end of another thread I have going about a bacteria problem Im having.  Figured it could use its own thread.  Apologies if Ive gone against basic ettiquette.

Thought my problem might be improper cleaning of the tupperware that Im colonizing my properly pasteurized coir/verm/gypsum sub in.  Using rye grain spawn prepared just like in RR's vids, except I PC for two hours to be safe.  When containers are colonized, I fruit the 'cakes' into a dialed in monotub, and have been having tremendous success...until the bacteria problem a couple of weeks ago.  The other thread I mentioned is all about the containers, so I wont go into that..but a few people mentioned that it might be my spawn (it ALWAYS smelled mushroomy and sweet..not the rank smells that would normally ID jars as bad.  But its a massive outbreak, so i have to look at all angles....so.....

Just contemplating how it might be the spawn thats the problem.  Ive had the MS culture going since early September.  I started from print->agar->jar>g2g 10jars...and from there spawned some jars, and g2g'd more...and so on and so on and so on.  So its been about 3 months now.

Im fairly confident with my sterile technique, and having the flowhood has really put my success rate way up.  So yes, im still considering that the problem is improper cleaning of the tupperwares..but......

Is it possible that the culture is just worn out due to senescence, and is simply more susceptible to bacterial contamination?

How old is too old for viable spawn?


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OfflineCMOS
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Re: How old is too old for viable spawn? [Re: Scarg]
    #19305241 - 12/20/13 09:25 PM (10 years, 2 months ago)

My problem with using older spawn is that its always a pain in the ass to break up into individual grains.  I believe its more likely to have bacteria growth after sitting around for a long time at room temp too. 

I usually try to break up old/questionable grain spawn jars a couple days before expanding, and checking to see if it recovers fairly quickly.  (like a day or two) If I do expand them to other grain jars, I give the empty jar a smell afterwards to check for obvious bacteria smells.

If it doesn't recover quickly, or break up easy enough, I just spawn them to some individual containers and let them do there thing.  (of course still do a smell/visual test before spawning.)


Ninja Edit : Just realized you have done a bunch of transfers from your original master spawn.  I would say your likelihood of a bacterial infection hitching along would definitely increase as you continue to expand.  More opportunity for bacteria to find entry, along with decreasing vigor of your original culture.  I usually try to start a new master line after about four transfers, but I don't know what the standard is.


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Edited by CMOS (12/20/13 09:27 PM)


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OfflineGuest F
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Re: How old is too old for viable spawn? [Re: CMOS]
    #19305288 - 12/20/13 09:36 PM (10 years, 2 months ago)

Quote:

CMOS said:
My problem with using older spawn is that its always a pain in the ass to break up into individual grains.  I believe its more likely to have bacteria growth after sitting around for a long time at room temp too. 

I usually try to break up old/questionable grain spawn jars a couple days before expanding, and checking to see if it recovers fairly quickly.  (like a day or two) If I do expand them to other grain jars, I give the empty jar a smell afterwards to check for obvious bacteria smells.

If it doesn't recover quickly, or break up easy enough, I just spawn them to some individual containers and let them do there thing.  (of course still do a smell/visual test before spawning.)


Ninja Edit : Just realized you have done a bunch of transfers from your original master spawn.  I would say your likelihood of a bacterial infection hitching along would definitely increase as you continue to expand.  More opportunity for bacteria to find entry, along with decreasing vigor of your original culture.  I usually try to start a new master line after about four transfers, but I don't know what the standard is.




I don't know for a fact, but I thought I remembered reading that 4 transfers was about the max; after then the spawn just becomes too weak to be practical.


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All cultivation posts are what I believe to be accurate. If I give ANY misinformation, please do not hesitate to correct me.


Edited by Guest F (12/20/13 09:36 PM)


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Invisibleazur
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Re: How old is too old for viable spawn? [Re: Guest F]
    #19305307 - 12/20/13 09:40 PM (10 years, 2 months ago)

Three transfers max. I use old spawn, like over 2 months old.  It's typically fine.


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OfflineCMOS
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Re: How old is too old for viable spawn? [Re: azur]
    #19305334 - 12/20/13 09:47 PM (10 years, 2 months ago)

I think its variable on max transfers.  Age of spawn, size of grain jars, and number of transfers all come into play.  For me I just generally won't go past four or so, and I try not to let them sit out fully colonized for long.

I end up throwing alot away (or making individual projects) and just starting a new line cause I never have room in my fridge to store them and they usually sit out for awhile.


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InvisiblePirateSwazey
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Re: How old is too old for viable spawn? [Re: CMOS]
    #19305442 - 12/20/13 10:20 PM (10 years, 2 months ago)

Hey dude.

I recently went through a similar situation back in September.

I believe at some point I used a master that had sneaky bacteria, and further spread the contamination to about 50 more jars from my 2nd generation of G2G transfers :thumbdown:

It sounds like this is what happened to you as well... especially since you feel like your sterile technique has improved w/ your new FH.

I just ended up tossing everything from that variety that I was working with and starting a couple new ones from scratch :shrug:


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OfflinecronicrFacebook
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Re: How old is too old for viable spawn? [Re: PirateSwazey]
    #19305774 - 12/20/13 11:58 PM (10 years, 2 months ago)

Drop a grain on agar you will your growth rate and or any contams, if you still got the og culture make a xfer and compare. No room for so on and so on how many jars did you do lol


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OfflineScarg
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Re: How old is too old for viable spawn? [Re: cronicr]
    #19305859 - 12/21/13 12:27 AM (10 years, 2 months ago)

Quote:

cronicr said:
Drop a grain on agar you will your growth rate and or any contams, if you still got the og culture make a xfer and compare. No room for so on and so on how many jars did you do lol




Tough to say how many I did.  300-400 after it was all said and done maybe?  From the first, I g2gd 10.  Then spawned 6 and made 40 from the remaining 4.  Then took 12 of those and did a batch of 120, while spawning the other 28..then just kept doing batches of 40 when the jars were ready.  So not sure if the contam came from unidentified bacteria in one or a couple of jars that just got propogated...or if the culture finally lost its gusto from being expanded too much.  It was a good 2 and a half months of success though.  Lots and lots of fruit..even just being multispore.

The part that confuses me is that I never smelled any bad smells.  Everytime I transfered anything, the smell test said I was good to go.

Luckily i started a new culture from spore->agar about two weeks ago, so hopefully Ill have another 100 jars or so relatively quick.  I suppose Ill just have to keep going back to spores sooner than I thought.


Another thing that confuses me is how the hell are you supposed to isolate strains from clones if it only takes a few months for a culture to get weak?


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OfflinecronicrFacebook
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Re: How old is too old for viable spawn? [Re: Scarg]
    #19305922 - 12/21/13 12:46 AM (10 years, 2 months ago)

Just keep your original culture so you dont have to keep going back to spores
And with cloning its the same thing, dont expand too much and save your culture so u have something to fall back on
And its not a time thing its how many cell divisions take place


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It doesn't matter what i think of you...all that matters is clean spawn

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Edited by cronicr (12/21/13 12:51 AM)


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OfflineStromriderM
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Re: How old is too old for viable spawn? [Re: cronicr]
    #19306443 - 12/21/13 05:01 AM (10 years, 2 months ago)

Quote:

cronicr said:
Just keep your original culture so you dont have to keep going back to spores
And with cloning its the same thing, dont expand too much and save your culture so u have something to fall back on
And its not a time thing its how many cell divisions take place




:whathesaid:


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