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OfflineHighHarles
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Registered: 07/07/18
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Jar Opinions Please
    #26471829 - 02/06/20 10:59 AM (4 years, 11 days ago)

I had a halfway decent write up of my knowledge of the flaws in my methods however my browser froze and it was cleared. Here's the summary

1 of 20 controlled jars (not inoculated) developed a bacteria after the pc cycle. It's my own dumbassary with the lids. Along with only using 1 layer of mp tape. Would this bacteria be the most prevalent in my environment being that it was the only one to develop?



Now these jars I've considered questionable for some time
Inoculated 12-20 w/ agar in a SAB (FH build in the nearish future, promise)
I numbered them to make it a little easier to keep organized an communicable.
Herrrreee we gooo
*mario noises*




Jars 6, 7, 8, 9, and 10 clearly producing metabolites. The majority of these look a little dull imo and have some similarity to that spotty bacteria developed in the control group.

What would you do with these jars? What's your opinion off looks?
I can take closer shots if need be, just ask


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InvisibleZifozonke
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Re: Jar Opinions Please [Re: HighHarles]
    #26471848 - 02/06/20 11:19 AM (4 years, 11 days ago)

Top fruit those jars...or..group the good jars together and the bad jars together do up a couple tubs and roll the dice...


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Invisiblealaskappalachian
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Registered: 10/22/19
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Re: Jar Opinions Please [Re: Zifozonke]
    #26471857 - 02/06/20 11:31 AM (4 years, 11 days ago)

looks like yeast contaminating the jar in your first pic.  That's a no-go.  Jar labled #2 looks a little spotty too, but maybe it's just the photo/my eyes playing tricks on me.  The rest I'd fruit, but I'd segregate as was suggested above (shoeboxes are a godsend for that very reason), in case you have problems that arise after fruiting.  Don't worry too much about metabolites, but segregate the jars that are presenting metabolites.


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OfflineTheApprentice
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Re: Jar Opinions Please [Re: alaskappalachian]
    #26471897 - 02/06/20 11:53 AM (4 years, 11 days ago)

if its bacterial, it most likely came from your sterile tech.

how long did you PC the jars for?

did you soak your grain over night?  did you rinse your grain and then dry it thouroghly before loading into the jars to PC?


--------------------


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OfflineHighHarles
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Re: Jar Opinions Please [Re: TheApprentice]
    #26473313 - 02/07/20 05:53 AM (4 years, 10 days ago)

Quote:

TheApprentice said:
if its bacterial, it most likely came from your sterile tech.

how long did you PC the jars for?

did you soak your grain over night?  did you rinse your grain and then dry it thouroghly before loading into the jars to PC?




I followed Bods easy af eat prep so it was a 2 hour cycle and 15 min vent at 15-18psi
-did I overlook the write up? I haven't seen any advisories stating I should rinse the grain, just boil/soak

-these grains dried over night in colanders and were removed the next day and stirred left on the counter for a few hours to rid of remaining excess

I havent opened these jars since the pc cycle. However caps are pretty loose coming out. Did reading and "shouldn't be an issue" so I'm 90% sure it's the original single layer of mp tape causing the issue.


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OfflineMrBovineJoni
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Registered: 11/28/19
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Re: Jar Opinions Please [Re: HighHarles]
    #26473341 - 02/07/20 06:45 AM (4 years, 10 days ago)

Some smaller amounts of metabolites are normal when jars reach full colonisation (especially if they have been colonised for a while) and does not always mean there is contamination.

Look at this post from RR explaining that metabolites can be fine : https://www.shroomery.org/forums/showflat.php/Number/20002949#20002949


Control jar can be a good point of reference but something growing on the grains is just a matter of time if you keep it long enough - even if the jar was never opened (but never say never - it can always happen!). When we  inoculate we want mycelium to take hold of all the "food" (grains) before other competing organisms do that first so we just try to give it a head start with grain sterilisation and sterile technique.

The other thing to consider is oats - people here have polarised love/hate relationship with that grain. Ones who have successes love it - ones who have had a failure or two consider them the worst (most contaminant prone, especially bacterial) grains ever; and there can be many factors that can cause this. Never touched them myself so can't provide personal experience.

In general I would say that there is something wrong with your jars if you used agar on them on 20th of December (so... 7 weeks ago) and they were not fully colonised within 4 weeks(unless you did not shake the jars at all at 20-30% colonisation; that would result in much slower colonisation times). However, you did not specify when exactly the jars were fully colonised so they might have been looking like this for weeks already? If that was the case in your place I would have spawned them a long ago already.

Just recently I put my first agar in my first ever rye jars (x10) on 22nd of January; shook about 8 days in and  3/10 are at 99% colonisation already and the rest should be there in the next 3-4 days; so definitely yours might have taken a while.


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OfflineHighHarles
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Re: Jar Opinions Please [Re: MrBovineJoni]
    #26473355 - 02/07/20 07:01 AM (4 years, 10 days ago)

There really hasn't been much trouble ime from the past. Just make em dryer. Only when using a syringe to grain. Too wet. Shit shoot. Less than 1/3 success rate.
(Just bought a whole ass fridge to streamline my process(like drying grains overnight, holding onto master jars etc))
I'd definitely recommend oats to starters cause it worked for me
They've just been hanging around for a while a couple grains have a bare spot, myc seems a little dull. I'll probably just empty out my 66 qts and fit them to a few shoeboxes ea
I plan on building a flow hood over the next month or two here so should make my work environment a little more ergonomic n less prone to contam
Plan on getting some grain bags going and using master jars to make the myc go a little further
Good luck on those transfers. This is my first a2g that turned out to be a shit shoot


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Offlinefootpath
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Registered: 07/16/19
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Re: Jar Opinions Please [Re: HighHarles]
    #26473369 - 02/07/20 07:14 AM (4 years, 10 days ago)

Quote:

stonesun said:
Well I didn't measure the the pore sizes,
instead took a piece of tape and mounted it on a slide.
Hydrated some P. cubensis spores and with an inoculation loop
mounted them on the tape.
The gaps are huge comparing to cubensis spores, which are ~12-17µm.
I circled the fibers with individual spores sticked to them.
Sorry for the kinda crappy images, didn't put a whole lotta effort in it...

100X


400X


400X





I've never used mp tape, but this kinda solidified that I don't ever want to.
Although tons and tons of people do get away with the double layer, I still prefer to put the little bit of extra effort and coin into modifying lids with SFDs


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OfflineHighHarles
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Re: Jar Opinions Please [Re: footpath]
    #26473417 - 02/07/20 08:07 AM (4 years, 10 days ago)

Quote:

footpath said:
Quote:

stonesun said:
Well I didn't measure the the pore sizes,
instead took a piece of tape and mounted it on a slide.
Hydrated some P. cubensis spores and with an inoculation loop
mounted them on the tape.
The gaps are huge comparing to cubensis spores, which are ~12-17µm.
I circled the fibers with individual spores sticked to them.
Sorry for the kinda crappy images, didn't put a whole lotta effort in it...

100X


400X


400X





I've never used mp tape, but this kinda solidified that I don't ever want to.
Although tons and tons of people do get away with the double layer, I still prefer to put the little bit of extra effort and coin into modifying lids with SFDs




Meh I had pretty good success with a single layer for sometime but I stumbled across this before. It's just easier to slap on a couple strips of tape as opposed to cutting tyvek discs n sealing them
I'll put the effort in in due time just doesnt seem worth a 5% contam rate battle at this moment

Next year I want to invest into a microscope so I can look at my cultures closer and play around observing different things like this


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