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Registered: 02/28/05
Posts: 16
Last seen: 11 years, 13 days
consistent contam after innoc
    #4096381 - 04/25/05 07:51 PM (12 years, 8 months ago)

Eight cakes became contaminated right at the inoculation points within days of innoculating. In the first set of 4 cakes, the needle was flamed with a lighter and cleaned off with a paper towel soaked isopropanol. Two different syringes were used (one species per jar), all contams (looks like the beginnings of blue-green mold). Maybe the paper towel wasnt sterile enough. Thus, in the next set of 4 jars the needle was only flamed in a clean-burning alcohol flame. The needle never touched anything but the innoc. holes. The needle was flamed each time and left to cool for a few seconds before inserting into each hole. The area around the holes was wiped down with ethanol to sterilize the lids futher. But it looks like there are still comtams occuring. In all cases the holes have had tape on since PCing them, so contams couldn't really get onto the top of the vermiculite layer and get pushed down into the substrate when the needle is inserted. There are still two of the eight jars left... still waiting to see if EVERY innoc. point gets contaminated to try to figure things out. Is it possible for the syringes to become contaminated, even if one of them hadn't been used before? The only other thing it could be is that the environment is chock full of mold in the air... This contamination spree never happened when inoculation was attempted at another location with one of the same syringes. Could someone tell me why this happened?

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Re: consistent contam after innoc [Re: sharppixy]
    #4096852 - 04/25/05 09:45 PM (12 years, 8 months ago)

Sounds like there is something in the syringe to me, did you happen to ever pull back on the syringe and pull air in? Those sterility procedures seem like enought to prevent contams, so im guessing its what you are shooting in. Good Luck on the last 2.

8 years now! lol

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Registered: 02/28/05
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Last seen: 11 years, 13 days
Re: consistent contam after innoc [Re: jamsandwich]
    #4098916 - 04/26/05 02:56 PM (12 years, 8 months ago)

Crap. The last two are contammed as well. There was some mycellium beginning to develop, but more blue green mold surrounding it. How did both syringes get contaminated? The syringes weren't pulled back after innoc., so it's hard to understand how contams got into the syringes... Next time there'll be new syringes and the old ones will still be around to test once more if they are really contammed (only on one or two cakes). Oh, and a more sterile environment too.

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Re: consistent contam after innoc [Re: sharppixy]
    #4101280 - 04/27/05 02:42 AM (12 years, 8 months ago)

Something to consider trying;  For the next batch, set aside two jars.  Keep one aside uninoculated.  Take the second and fake inoculate:  if you're working with two syringes, stick each needle in two of the four holes but don't push out any liquid.  Flame the needle and do everything the same as the real jars.  If the air is just really filthy [or the needle], the fake inoculated jars would be just as likely to contam as the real jars.  If only the real jars contaminate, the syringe fluid is likely mold-ridden. 
It's not a guaranteed answer, but probably worth trying after a 100% loss :frown:  The syringes can be replaced, but it would be good to know if the environment is actually the problem.

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Mushrooms, Mycology and Psychedelics >> Contamination Forum

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