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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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How can this syringe be saved?
#19185572 - 11/25/13 06:23 AM (10 years, 2 months ago) |
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There is a spore syringe that was purchased that repeatedly produces what looks like a strange contam when using on agar. Within a day, the areas where the water has touched/rolled on the agar will turn a bright pinkish red almost resembling dilute blood, then it seems to gelatinize(is that a word?) and take on a beige color. After that it seems to stop progressing. Every plate made with the syringe turns out this way no matter how sterile the technique making me think it's a problem with the syringe itself. So, wanting to be a proper agartista, how would someone go about growing anything and then separating in this environment? There are several plates new plates with this now and there are visible spores present. Whether or not the spores will germinate in the first place is the big question since they all are in this gook. What to do if they manage to grow is another.
Perhaps a hot agar bath? But do the spores need to germinate first? Or maybe a antibiotic agar will need to be made? Maybe there is a way to remove this pinking fluid before it gets a foothold atop the spores and agar?
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SamhainJ
I wanna rock out, in my dreams
Registered: 05/16/04
Posts: 1,002
Loc: USA
Last seen: 8 years, 7 months
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Re: How can this syringe be saved? [Re: NOS4A2]
#19185595 - 11/25/13 06:35 AM (10 years, 2 months ago) |
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Wouldnt it just be worth it to toss the spike if its contamed aand make or buy a new one?
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SupaThaRipper
Genetics Hoarder



Registered: 09/02/13
Posts: 1,504
Loc: USA
Last seen: 1 hour, 2 minutes
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Re: How can this syringe be saved? [Re: SamhainJ]
#19185648 - 11/25/13 07:03 AM (10 years, 2 months ago) |
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I'm still a noob so this may not be the best way to go about it, but what I would do is inoculated a grain jar and hope that some decent mycelium forms out of it. Then, grain to agar and try to isolate from contamination. What type of mushroom is it btw?
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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You may have a good idea Supa. Maybe if the water were allowed to drain away from the spores they'd have a better chance. I was told it was B+. The first time it was opened it got stuck into the nail bed of a finger accidentally, so I don't want to discredit this well known vendor for sending a bad sample. However, a different vendor will be chosen next time. The less that has to be ordered the safer, so if something can at least be gained from this syringe the better.
Not to mention as stated by more experienced cultivators before, a good cultivator would be able to transfer the contam away. I like the grain idea which might be tried but agar is made for this sort of situation. Maybe if I can get a pic of the sample it will spark some ideas...
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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Re: How can this syringe be saved? [Re: NOS4A2]
#19186366 - 11/25/13 10:49 AM (10 years, 2 months ago) |
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Can a syringe be PC'd to a certain temp killing bacteria but sparing the spores?
Or perhaps the contents could be removed, treated, quickly dried out in a shot glass, then rehydrated with a peroxidated solution?
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Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 3 years, 1 month
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Re: How can this syringe be saved? [Re: NOS4A2]
#19189306 - 11/25/13 10:03 PM (10 years, 2 months ago) |
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What you *can* do is sterilize a shot glass full of water, shake the syringe thoroughly and inject a single drop of spore solution into the glass. Swirl to mix. Then, with a loop or other inoculating device, remove a tiny quantity of that water and streak it in a zigzag pattern across your agar.
What you *should* do is call up the provider of the syringe and tell him that under a microscope it appears the syringe is contaminated with some sort of bacteria, and that it's unsuitable for microscopy.
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,855
Last seen: 14 hours, 39 minutes
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Re: How can this syringe be saved? [Re: NOS4A2]
#19189554 - 11/25/13 10:47 PM (10 years, 2 months ago) |
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You could try antibiotic agar. Don't know how much it would help though if it's that heavily contaminated. Be sure you're only a tiny drop to incoulate the agar. It's easy to over do it and no mycelium if there's too much moisture.
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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Re: How can this syringe be saved? [Re: Kizzle]
#19191142 - 11/26/13 10:00 AM (10 years, 2 months ago) |
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'What you *can* do is sterilize a shot glass full of water, shake the syringe thoroughly and inject a single drop of spore solution into the glass. Swirl to mix. Then, with a loop or other inoculating device, remove a tiny quantity of that water and streak it in a zigzag pattern across your agar.'
good one
'What you *should* do is call up the provider of the syringe and tell him that under a microscope it appears the syringe is contaminated with some sort of bacteria, and that it's unsuitable for microscopy.'
like I said after sticking it into a finger before even the first innoc with that syringe, it can't be honestly argued that it was contammed before that. Saving a buck just isn't that important in this case. Lesson learned.
'You could try antibiotic agar. Don't know how much it would help though if it's that heavily contaminated. Be sure you're only a tiny drop to incoulate the agar. It's easy to over do it and no mycelium if there's too much moisture. '
I guess this is as good a time as any to learn to make antibiotic agar. And yes, scraping some spores onto agar is a lot cleaner process than dripping them from a syringe. I really don't think using a syringe for agar will happen in the future...
I was kinda hoping someone like FrankHorrigan with a considerable more experience woulda chimed in by now. Especially when he's chastising people for not working with contammed cultures and just tossing them.
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