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OfflineIdoThisforResearch
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old spore syringes failed! *PICS*
    #21944379 - 07/14/15 10:42 PM (8 years, 6 months ago)

So a guy from the job tried to knock up some BRF Jars with spore syringes that had been in the fridge for 18 months....failed miserably! Judging by the pics, does it look like the syringes just went bad? He had heard of others having success with old syringes, so was understandably disappointed when ALL of his jars failed!


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Offlinematsc
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21944442 - 07/14/15 10:58 PM (8 years, 6 months ago)

Thats contamination alright. However, its difficult to tell if the contamination was in the syringes all along, or if the inoculation technique was at fault. Its possible they were poorly capped and got some gunk in them, or the needle wasnt thoroughly flamed before hand.

In any case, theyre toast. Either toss them in a bucket of bleach water to soak, or take them outside to clean them out. Or both. Just dont open them indoors as they are now.


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21944555 - 07/14/15 11:13 PM (8 years, 6 months ago)

The fridge is a bad place to store spores or cultures. Completely ridden with contams, the syringes could have been made in a nonsterile environment too.

People have a lot of issues with contaminated prints and syringes :shrug:


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OfflineIdoThisforResearch
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21945958 - 07/15/15 06:04 AM (8 years, 6 months ago)

Don't worry, they have already been tossed! Lol
They were securely capped the entire time that they were stored (in an airtight double sealed ziplock bag).
Needle was wiped with alcohol + flamed sterilized according to tek procedures between each jar.
The spores were from a reputable source and the guy had success on two other occasions prior (with spores from the same supplier). Thats why he's thinking that maybe they just died in the fridge over time?
Thanks for the quick responses...any other thoughts?


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OfflineFreeWorldOrder
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21946391 - 07/15/15 08:46 AM (8 years, 6 months ago)

Syringes should keep for quite a while when properly stored. Sounds like they were stored okay. I keep mine in the fridge in a ziplock and have never run into any problems so far. Prints should keep for years.


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Re: old spore syringes failed! *PICS* [Re: FreeWorldOrder]
    #21946481 - 07/15/15 09:13 AM (8 years, 6 months ago)

the syringes were dirty, you can tell because contamination begun right where you inoculated.
if it was indeed from a reputable vendor or even better a shroomery sponsor
just contact them and say the spores werent viable under a scope. decent vendors will replace your syringe.


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Offlinematsc
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Re: old spore syringes failed! *PICS* [Re: spacechildo]
    #21946498 - 07/15/15 09:18 AM (8 years, 6 months ago)

Even if the spores died in the fridge, that wouldnt have changed them into the various molds you see in those jars. You would have just seen nothing happening if that were the case. Some nastyness got into them at some point, it happens.


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OfflineIdoThisforResearch
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Re: old spore syringes failed! *PICS* [Re: matsc]
    #21946591 - 07/15/15 09:58 AM (8 years, 6 months ago)

gypsum was also used for the first time on these jars and wasn't sure if it was the right stuff to use, but it was the only thing at the store that said "gypsum" on it....any thoughts on this stuff? It was like rock hard pellets that had to be crushed with a hammer down into dust to mix with the sub.



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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21947194 - 07/15/15 12:44 PM (8 years, 6 months ago)

Quote:

Even if the spores died in the fridge, that wouldnt have changed them into the various molds you see in those jars. You would have just seen nothing happening if that were the case. Some nastyness got into them at some point, it happens.




Yep, i also have doubts that bacteria can not permeate through pores in plastic after such a large amount of time. Microscopic dust permeates the bags when you put cannabis in them so dogs eventually smell it, just a matter of time. Why couldnt bacteria or spores enter?

Theres so many different types of contams its hard to believe for one second that they were clean from the start. A fridge is normally all bacteria, you look like you have lots of different molds and bacteria. It also all started at the injection site so it wasnt fom the substrate.

Quote:

Prints should keep for years.




Yeah but only retain like 1/10,000th of the viability.

Gypsum pellets wont cause issues if you sterilize properly. Those do look a little dark and large though, is it supplemented with something?


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Offlinespacechildo
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21947214 - 07/15/15 12:47 PM (8 years, 6 months ago)

1/10000th?

2-3 yrs old prints germinate just fine for me, 5 yrs old take a cpl weeks but grow fine still.


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: spacechildo]
    #21947392 - 07/15/15 01:28 PM (8 years, 6 months ago)

Quote:

I've ran several tests with the microscopes to determine spore viability. A brand new print will give about 1 spore in 100 that will germinate. After two weeks, that drops to about 1 spore in 500. At one month since taking the print, less than 1 spore in 1000 will germinate, and after one year, you're doing good to get 1 spore in 10,000 that is still viable and it drops even faster after one year.

The thing to remember is take clean prints and store them properly. An old print will still work, but remember that you'll need to use many more spores to get the few that will germinate. Remember, when you use more spores, you also risk using more contaminant spores that may be hitch hiking along for the ride. Thus, fresh prints are better.
RR




If you have good technique it will still work but not nearly the same as spores less than a year old.

If you can germ a 4-5year old print or syringe, you should be proud of your technique :thumbup:


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Offlinespacechildo
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21947536 - 07/15/15 02:05 PM (8 years, 6 months ago)

IDK how he counted that, but even if its true 1/10000 still alive is probably way more than I need still. if you can see a clump of spores you have an insane amount of spores.

I thought for a second you meant 1/10000th the chance to get anything to germinate.

It has nothing to do with technique, just scrape and wait :shrug:

he also says spores survives the longest in a syringe, 10 yrs easily, but survives shorter on a print.
My exp is the opposite. I dont quite remember but he may have gone back on that
and said prints die because some people store them in books...


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: spacechildo]
    #21948321 - 07/15/15 04:52 PM (8 years, 6 months ago)

I was interested in how he determined the viability of the spores as well. IME and my friends experience we always seem to have issues when they hit the 4 year mark. :shrug:

Maybe it depends on storage conditions and the  water quality of the spore water. I always had prints last longer too :confused:


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Offlinematsc
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21949140 - 07/15/15 07:59 PM (8 years, 6 months ago)

I've germinated 16 year old spores, though of a different species and store very carefully (Aspergillus flavus, stored on desiccant in a freezer). I've also grown successfully from spores stored in liquid that were 12 years old. A normal spore print on foil, though, that could depend on a ton of variables (humidity, temperature, spore density, etc etc).


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: matsc]
    #21949206 - 07/15/15 08:16 PM (8 years, 6 months ago)

It must be storage issues rather than spore longevity.

:themoreyouknow:


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OfflineIdoThisforResearch
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21949357 - 07/15/15 09:02 PM (8 years, 6 months ago)

To answer your question from earlier Toadstool, I don't think the gypsum was supplemented with anything. Heres what the bag said...kinda hard to read though:


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21949378 - 07/15/15 09:08 PM (8 years, 6 months ago)

I think it is gypsum but processed differently than the stuff i am used to. Heres a picture of what i use:



Maybe mine is drier?


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InvisibleDr.Satan
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21949406 - 07/15/15 09:16 PM (8 years, 6 months ago)

What is the point of adding gypsum? Is it suppose to be added into the verm and BRF or is it suppose to replace one of them?


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: Dr.Satan]
    #21949448 - 07/15/15 09:28 PM (8 years, 6 months ago)

It supplies much needed calcium and sulfur.

The added calcium increases the strength and durability of vacuoles as they swell inside the aging hyphae. I forget what paper i had read it in but supplementing with gypsum can have a significant effect on yield. As much as about 10-20% with agaricus bisporus.


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InvisibleDr.Satan
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21949454 - 07/15/15 09:31 PM (8 years, 6 months ago)

Quote:

Toadstool5 said:
It supplies much needed calcium and sulfur.

The added calcium increases the strength and durability of vacuoles as they swell inside the aging hyphae. I forget what paper i had read it in but supplementing with gypsum can have a significant effect on yield. As much as about 10-20% with agaricus bisporus.




I'm definitely going to look into that, it sounds like it's worth a shot. Thanks for the info. I learn something new everyday on here xD


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: Dr.Satan]
    #21949567 - 07/15/15 09:58 PM (8 years, 6 months ago)

I know how that is :lol:

Just this thread has taught me so much on spore aging :thumbup:


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OfflineIdoThisforResearch
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21949599 - 07/15/15 10:06 PM (8 years, 6 months ago)

Hmm, yea maybe it was a different kind of processing.
Is yours really easy to crush or crumble to mix with? Bc this stuff was HARD AS ROCKS! Literally was beating on it with a hammer for a couple minutes, and truth be told most of the pellets were still intact! Lol
I ended up just using the dust that had collected in the bottom of the bag.


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OfflineIdoThisforResearch
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21949605 - 07/15/15 10:07 PM (8 years, 6 months ago)

And also Toad, where do you get your gypsum? Bc this was the only stuff that was at the store....


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21949679 - 07/15/15 10:31 PM (8 years, 6 months ago)

Same with mine, i dont even attempt to grind it up.

I got it from a friend who i believe got it at Lowes in the gardening section.


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OfflineIdoThisforResearch
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Re: old spore syringes failed! *PICS* [Re: Toadstool5]
    #21949756 - 07/15/15 10:56 PM (8 years, 6 months ago)

Ok cool. So do you just mix it straight up as it is? Or do you also try to pound it with a hammer? Just curious, bc I thought I had read somewhere that gypsum was really easy to crumble and mix, but maybe its just different depending on where you get it?

And anyways, back to the original topic, Im pretty sure thanks to yours and everyone else's input, that the gypsum had nothing to do with the contam....but just bad/old syringes! Thanks for all the help!


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InvisibleToadstool5
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Re: old spore syringes failed! *PICS* [Re: IdoThisforResearch]
    #21950026 - 07/15/15 11:41 PM (8 years, 6 months ago)

I just mix as is because the stuff i have is too hard to break up. I have seen a few people dissolve it in water and use the gypsum water to hydrate their brf/verm or grains. Usually in my experience gypsum is almost always very hard.

No problem, we like to help :thumbup:


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