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InvisibleSky Walker
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Registered: 02/11/12
Posts: 605
Thoughts on Contam in BRF Jar (with pic)
    #23616294 - 09/05/16 09:01 PM (7 years, 4 months ago)

First time noob here.  Followed the PF Tek as closely as possible and was thoroughly sanitary throughout the entire process, especially inoculation. Only thing I did differently was I actually PCed my jars. All jars have been sitting in my closet at about 78-79 degrees for about 12 days now.  Four out of eleven jars are finally showing decent growth and the rest have no signs of germination.  The four with growth were definitely a bit stalled.

Getting to the point, 1 of the 4 jars with growth is showing a sign of contamination and it is the only jar thus far with any sign of contam.  The white spot it clearly visible in the picture below and has not grown much or changed colors since I first noticed it about four days ago.  Thought I'd watch to see what happens and at first was not sure if it was a contam, but now it is clear as the myc has converged and started to grow around it.

I've searched the forums and haven't found anything that looks quite like this, at this stage of growth, and I'm not sure if it's salvageable. I know cobweb is super rare in BRF jars, but it may be something similar.

I figured some may be interested and any thoughts and/or advice is much appreciated.  Also the only thing I can conclude went wrong with my low germination rate is that I didn't shake the syringe enough during inoc. I shook the hell out of it before placing it inside of my SAB, but should I be shaking during the inoc process as well?
THANKS!!



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OfflinePsilicon
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Sky Walker]
    #23616426 - 09/05/16 09:47 PM (7 years, 4 months ago)

Definitely not cobweb.  It's probably a pin mold or something, and likely just the cost of business working with spore syringes.  Your prep looks good and I have confidence you'll have better luck with your other jars.


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InvisibleSky Walker
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Psilicon]
    #23617790 - 09/06/16 11:03 AM (7 years, 4 months ago)

Thanks Van.  Any thought on my very low germination rate.  I believe it was a little less than 14% of all ports on my jars actually germinated.

Do you think this was a consequence of not shaking the syringe enough during inoculation?  I will say that I ordered my spores from a sponsored vendor with a good reputation, so it must have been something during my procedure.


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InvisibleMad Season
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Psilicon]
    #23617974 - 09/06/16 12:09 PM (7 years, 4 months ago)

Quote:

van der griegen said:
Definitely not cobweb.  It's probably a pin mold or something, and likely just the cost of business working with spore syringes.  Your prep looks good and I have confidence you'll have better luck with your other jars.



Where is this pin mold? :confused:. I mean I'm zooming in a ton here and that just looks like tomentose myc to me. :/


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InvisibleSky Walker
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Mad Season]
    #23618103 - 09/06/16 12:46 PM (7 years, 4 months ago)

Ah that's interesting. Hadn't considered this as a possibility.  Not to immediately assume its a contam, but I was suspect because of the very different appearance in the texture of the two growths and difference in texture.  They also appear to be competing, or at least to my untrained eye.  I have circled the area of concern in the image below.  See how the obvious myc growth appears to be avoiding the lighter/smaller growth above it?  Is this not indicative of a competing contam?  That said, its been about five days since I noticed the different spot and two days since they began to converge and no color change in either one.

Thanks for y'alls responses, really just more curious and interested in learning how to identify these things.  If its good, GREAT, if not, oh well I'll toss it.



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InvisibleMad Season
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Sky Walker]
    #23618284 - 09/06/16 01:32 PM (7 years, 4 months ago)

I guess it looks similar to aspergillus or something before sporulating. I personally would just let it sit and see what happens. I see no reason to toss yet tho. It doesn't look at all like a pin mold or cobweb.


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Improve your sterile techniques! (A comprehensive guide to agar)
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OfflineThe Mycologist
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Mad Season]
    #23618292 - 09/06/16 01:36 PM (7 years, 4 months ago)

Yeah might just be mycelium


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InvisibleSky Walker
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Mad Season]
    #23618423 - 09/06/16 02:15 PM (7 years, 4 months ago)

Wicked!  Thank you guys.  Had no idea the mycelium could even take variant forms.  Like I said, even if it turns out to be contaminated, I appreciate the knowledge and guidance.


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Offlinemrmazdarx9
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Sky Walker]
    #23618441 - 09/06/16 02:19 PM (7 years, 4 months ago)

If it changes colour then worry


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OfflineMushierage
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Sky Walker]
    #23618567 - 09/06/16 02:57 PM (7 years, 4 months ago)

Quote:

skywalker444 said:
Wicked!  Thank you guys.  Had no idea the mycelium could even take variant forms.  Like I said, even if it turns out to be contaminated, I appreciate the knowledge and guidance.




Yeah, just let it go.  It's entirely possible that it's just Myc and it's fine.


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OfflinePsilicon
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Mushierage]
    #23619932 - 09/06/16 09:36 PM (7 years, 4 months ago)

Still looks sketch to me, especially when considering how it looks like the more rhizomorphic growth is avoiding it.  But I agree to hang on to it and keep an eye on it.


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Agar - what, why and how?  Everything a beginner needs to know.
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InvisibleSky Walker
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Re: Thoughts on Contam in BRF Jar (with pic) [Re: Psilicon]
    #23633309 - 09/10/16 05:51 PM (7 years, 4 months ago)

UPDATE:

So the jar from my OP has continued to thrive, with no progressing sign of contam.  The rhizo myc has mostly grown around the spot of concern and somewhat through it.  The rhizo myc is definitely growing faster. I'm starting to think this is in fact a small spot of tomentose myc of some varying genetics that were present in the MS inoc.

Here is an updated picture of the jar in OP


So that's the good news. The bad news is that I had a variety of other contams in three different jars.  I noticed that the cakes with myc growth had more of a tendency to shrink than the other cakes with no growth of any kind.  Some condensation appeared a few days ago in all of the jars with growth.  One day later some obvious contams popped up in three of those jars, some of which were extremely aggressive and seemingly ran through the jar in a single day.

These are those three contam jars.  The contams varying in color from white to black to green.


Out of 11 jars I am now left with only 2 that continue to appear viable. This has definitely been a learning curve, but one that I have learned a lot from.  I have many ideas of where I went wrong and hope to have better luck next go around.


Edited by Sky Walker (09/10/16 07:53 PM)


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