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OfflineEp1429
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Bacterial contams on agar? * 1
    #25619114 - 11/16/18 02:58 PM (6 years, 2 months ago)

I got four syringes of B+ from an online vendor, and after a 50%-65% success rate with BRF cakes and WBS using three of them, I decided to test the fourth on agar to see if the syringes were dirty, since I’ve never had that low a success rate in the past.

I poured several MEA plates, inoculated 6 with the syringe, 6 with spores from a print I made, and four with tissue from a biopsy for cloning. I have nine poured plates that I was going to use for transfers, but which are acting as my controls for this test.

All six of the syringe plates look like this, which I am assuming is bacterial growth, the same as what I’ve seen in the past in a vet lab:





In that one, the bacterial slime actually flows across the surface of the plate.



That one is the only one of the six that looks like it has any mycelium at all.

For comparison, one of the spore print plates. None of them have any contams yet, nor mycelium. They’re clean from the white slime, as well as the control plates I’ve set aside.



Am I correct in saying that is bacterial contamination?
Six for six on the syringe plates, nothing like that on the clone plates, print plates, or controls. Am I safe in assuming my syringes were FUBAR on arrival?

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OfflineCzM8
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Re: Bacterial contams on agar? [Re: Ep1429]
    #25619300 - 11/16/18 04:26 PM (6 years, 2 months ago)

It could be sterile technique. My first syringe to agar contamed like that, but I also put them on grain and they were okay.

For agar and syringe, I don't even use a needle.


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OfflineCzM8
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Re: Bacterial contams on agar? [Re: CzM8]
    #25619301 - 11/16/18 04:27 PM (6 years, 2 months ago)

Also, if your going to do agar, just get a print. Really, its much easier ime.


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OfflineEp1429
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Re: Bacterial contams on agar? [Re: CzM8]
    #25619334 - 11/16/18 04:40 PM (6 years, 2 months ago)

Quote:

CzM8 said:
Also, if your going to do agar, just get a print. Really, its much easier ime.




I have three prints made from my first group of shoeboxes. I’ve got my eye on two subjects in the current batch of shoeboxes I have going. Prints won’t be a problem. Like I said, I’ve got six plates with spores on them already, and no contams yet.

But I wanted to learn how to work with agar and to test my hypothesis about the syringes, and I’ve already learned a lot about the process in my first attempt.

If I can assume the syringe is the culprit, I’ll inoc some more BRF cakes and see what I can get out of it, then switch entirely to agar and my prints.

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InvisibleShroomspective
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Re: Bacterial contams on agar? [Re: Ep1429]
    #25619441 - 11/16/18 05:29 PM (6 years, 2 months ago)

Top 3 pics all look bacterial, and the last one doesn't look like mycelium either

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OfflineEp1429
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Re: Bacterial contams on agar? [Re: Shroomspective]
    #25619559 - 11/16/18 06:39 PM (6 years, 2 months ago)

Correct!  The last one has no mycelium visible yet, though you can see the zigzag from my loop.  That is a spore print plate, and they were knocked up 49 hours ago with the rest of them.

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Re: Bacterial contams on agar? [Re: Ep1429]
    #25620078 - 11/17/18 02:51 AM (6 years, 2 months ago)

In that case the last one is looking good!

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