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sk33mz
Mycoaddict

Registered: 12/15/10
Posts: 31
Last seen: 12 years, 8 days
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Spore death in route?
#14755253 - 07/12/11 03:57 AM (13 years, 6 months ago) |
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I ordered four different syringes from -----, and they threw in the free one for being a member. TC, GT, APE, PR, and CR. I injected four jars of WBS for each strain a month ago today using Doc's WBS tek which I have had great success with multiple times in the past. They show zero growth.
After thinking my procedure over the only thing I could come up with was maybe I may had gotten the moisture content wrong the first time around, so I prepared more WBS allowing some to dry longer and some to remain a little more moist. I then varied these jars so that each strain got two of the drier jars, and two of the more moist and injected four jars for each strain again. These jars are on 10 days today with zero signs of growth in any of the jars. I know they could still show but I would expect a spot or two on one of the twenty jars by now.
---- has always been great and I have had great success with them in the past, so I don't want to believe the spores are bad. However I have always had decent to great results using Doc's WBS tek and even if it was my error, I would think one of the 40 jars would show at least something at this point. Then I thought about the temperatures of the spores being shipped. It has been steadily above 100 degrees here for the past month or two. The house is kept at 75F or so, so that is not the issue. However I am sure the back of the delivery trucks that brought the spores could hit very much above 100F and I have read that spore death can occur at 106F (not sure if that is accurate). Is there a possibility that the spores were ruined in transit?
I started some LCs with all of them to see if the spores would even germinate that, however it is still to early to tell from them. I went from about a 98% success rate to straight zero and I can't figure out for the life of me why. 
*edited to follow rules*
Edited by sk33mz (07/25/12 11:53 PM)
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Rewindicus
Silly Goose



Registered: 06/05/11
Posts: 5,491
Last seen: 3 years, 2 months
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Re: Spore death in route? [Re: sk33mz]
#14755279 - 07/12/11 04:13 AM (13 years, 6 months ago) |
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my syringes from sporeworks have all worked fine. whats the temp of the area your keeping them in for incubation? is it possibly really cold? like 60? i had mine in a 74 degree closet and had growth with my last 2 batches after 3-5 days. thats the only thing i can think of in my limited knowledge or is it possible you live somewhere so hot that in transport there was an over 106 degree temperature situation and maybe the spores died?.....because like you say even if moisture content was jacked you should have had SOME growth somewhere.
-------------------- “Be who you are and say what you feel because those who mind don't matter and those who matter don't mind.”- Dr. Seuss
"Too much of a good thing, can be wonderful!" - Mae West
"If you have nothing nice to say about anyone, come sit next to me."
- Alice Roosevelt Longworth
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sk33mz
Mycoaddict

Registered: 12/15/10
Posts: 31
Last seen: 12 years, 8 days
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Re: Spore death in route? [Re: Rewindicus]
#14755351 - 07/12/11 04:56 AM (13 years, 6 months ago) |
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Thanks for the quick reply. I should have included more background info.
The area they are kept in stays at about 75F. Very sterile procedure, including glovebox. Soak and very quick simmer, rinsed, etc. PC'ed at 15 psi for 90 minutes. Cooled overnight. Filtered with tyvek from the painting coveralls. No incubation chamber. Ambient light 12/12. I usually have great success with everything. Spores always from Sporeworks and WBS always done the same. Was just now about to venture into LC and agar, but that is off topic and now I don't have many spores left to do it. 
I really don't think its bad spores from sporeworks, but the procedure is exactly the same and I have ZERO growth in 40 jars. 20 a month old and 20 ten days old. Like I said 98% success to zip.
It has been steadily 100+F here daily for at least a month. I get in the car and the thermometer says 107F all the time just from sitting in the parking lot for a few hours. So I know the back of the delivery trucks must be AT LEAST 10 degrees warmer than outside. I hope I didn't get 5 syringes of toasted spores.
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Rewindicus
Silly Goose



Registered: 06/05/11
Posts: 5,491
Last seen: 3 years, 2 months
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Re: Spore death in route? [Re: sk33mz] 1
#14755367 - 07/12/11 05:06 AM (13 years, 6 months ago) |
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this is actually a post from sporeworks from 3 years ago about heat and shipping...
http://www.shroomery.org/forums/showflat.php/Number/7265596#7265596
"Howdy folks,
It looks like summer is in full force this week, with Heat Advisories posted for a good portion of the country. I thought it might be a good idea to take a second and remind people that special considerations should be taken when ordering sensitive biological material during periods of especially high heat.
Spores will suffer thermal death if exposed to temperatures above 120F. Sporeworks highly recommends taking advantage of expedited shipping, express mail, or air services to minimize the possibility of high heat exposure during transit.
We find that utilizing services such as Priority Mail or Express Mail from the postal service will greatly reduce the potential for exposure. Both these services are available from our site by selecting the "Other Carriers" option during chekcout.
For those of you who prefer UPS, we offer Next Day Air, Second Day Air, and 3 Day Select options as expedited shipping. This will help minimize the time your package spends in over the road trucks during transit.
Keep in mind that Sporeworks guarantees all our material, even if it is rendered unstable from heat exposure. We wanted to alert folks to the available shipping methods available, as the old saying goes "an ounce of prevention is worth a pound of cure." So while we will send you replacements if your samples arrive unstable, it's worth considering these special circumstances during really hot spells and preventing the damage in the first place. "
--------------------------------------------------
BUT! they do say they guarantee all materials so maybe you should shoot them and e-mail and see about replacements?
-------------------- “Be who you are and say what you feel because those who mind don't matter and those who matter don't mind.”- Dr. Seuss
"Too much of a good thing, can be wonderful!" - Mae West
"If you have nothing nice to say about anyone, come sit next to me."
- Alice Roosevelt Longworth
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sk33mz
Mycoaddict

Registered: 12/15/10
Posts: 31
Last seen: 12 years, 8 days
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Re: Spore death in route? [Re: Rewindicus]
#14755417 - 07/12/11 05:33 AM (13 years, 6 months ago) |
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Wow, nice find. I can't believe I didn't find that while searching for a week. Thanks.
I could easily believe that it would be over 120 in those trucks out here after a long ride. Its good to know Sporeworks still guarantees them, I didn't consider the heat being an issue until after nothing worked or I would have gone express shipping. I'll give the second batch of jars another week and if I see nothing in those or the LCs, I'll hit up sporeworks and hope for the best. Sucky learning experience but eh, something had to go wrong sooner or later. Oh well, it made me stop lurking and start posting.
Thanks again. If I could give you rep or shrooms or whatnot I would but I think I need more posts or I'm just to baked to figure it out.
Edited by sk33mz (07/12/11 05:36 AM)
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Rewindicus
Silly Goose



Registered: 06/05/11
Posts: 5,491
Last seen: 3 years, 2 months
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Re: Spore death in route? [Re: sk33mz]
#14755436 - 07/12/11 05:40 AM (13 years, 6 months ago) |
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im sure you already know but if you have experience from before....no spore prints? should prob get some made next time you can that way you never get stuck with a hot summer heatwave situation again. =)
-------------------- “Be who you are and say what you feel because those who mind don't matter and those who matter don't mind.”- Dr. Seuss
"Too much of a good thing, can be wonderful!" - Mae West
"If you have nothing nice to say about anyone, come sit next to me."
- Alice Roosevelt Longworth
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sk33mz
Mycoaddict

Registered: 12/15/10
Posts: 31
Last seen: 12 years, 8 days
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Re: Spore death in route? [Re: Rewindicus]
#14755460 - 07/12/11 05:54 AM (13 years, 6 months ago) |
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Yeah, I've got some EQs and Hawaiian prints tucked away. Didn't want to have to go out and buy new syringes and needles cause I hate doing it, but now I have some empty ones to re-sterilize and reuse. 
Was excited to get all the new strains going especially the Albino PEs. Was gonna venture into agar and LCs with these, but I suppose I could bust the prints out to hold me over. Hopefully I was sterile enough. It was done correctly, I believe. But my only other attempt to make a syringe didn't work. That was long long ago though.
Edited by sk33mz (07/12/11 05:56 AM)
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