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OfflineMagicFingers
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Registered: 04/30/15
Posts: 214
Last seen: 7 years, 5 days
Contamination Issues....again
    #23812534 - 11/08/16 12:12 PM (7 years, 3 months ago)

I've had a number of projects in the past and recently I've been having a lot of failures due to contamination.  The most common issue I've seen has been bacteria. I do the normal 12-24 hr soak and rinse the grain very good before I let sit covered and load the jars and PC for 90m.

I know my challenge is I have to use a plug in  burner. This has caused issues with getting the PC hot enough. But if I PC too long the grain starts turning to Mush. I work around this I tried getting a larger for efficient plug in burner. Ends up that induction doesn't work on my PC....

Here is an example of some WBS I recently inoculated. As soon as I saw some growth I shook the jars. This is a bout a week after that. I've had this issue a lot. Obviously, the jars that had super fast colonization is looking at maybe a cotton like mold? I also see bacteria as well. The only jar that looks half decent is the middle one. Which is starting to show Mycelium growth. But my guess is bacteria will win that one out too.

I use a SAB. I cover everything with either an isopropal alcohol or bleach water.


Thoughts?



My lids are made with High Temp RTV silicone and synthetic filter discs cut and siliconed on to the lid. Maybe the filter discs are getting wet causing bacteria to seep through? *just a thought* Maybe when I am doing my inoculation in the SAB I should be careful how much alcohol or bleach water I get on the SFD?

Either way, I've been getting a bit irritated with these issues. So I've decided to get a few rye berry spawn bags from a local vendor.  I hope these can help me get some good projects.

Please no hate. Just trying to get some constructive criticism.

Thanks


--------------------
Live the Life you Love!
-MagicFingers:mushroom2:

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InvisibleMad Season
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Registered: 09/16/12
Posts: 12,666
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Re: Contamination Issues....again [Re: MagicFingers]
    #23812570 - 11/08/16 12:20 PM (7 years, 3 months ago)

And you use agar? It will tell you everything you need to know. How good your sterile techniques are, and if your syringes are contaminated.


--------------------
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Improve your sterile techniques! (A comprehensive guide to agar)
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OfflinePNW_Shroomer
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Registered: 11/08/16
Posts: 42
Loc: Pacific Northwest
Last seen: 7 years, 3 months
Re: Contamination Issues....again *DELETED* [Re: MagicFingers]
    #23812583 - 11/08/16 12:24 PM (7 years, 3 months ago)

Post deleted by PNW_Shroomer

Reason for deletion: ...


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InvisibleMad Season
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Registered: 09/16/12
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Re: Contamination Issues....again [Re: PNW_Shroomer]
    #23812592 - 11/08/16 12:27 PM (7 years, 3 months ago)

That's funny because I've had more issues with larger grains like rye and oats than I have had with smaller grains like wbs and Milo. With wbs I just soak for 12 hours, then load in the PC. No simmering at all. They come out beautifully.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.

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OfflinePNW_Shroomer
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Registered: 11/08/16
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Re: Contamination Issues....again *DELETED* [Re: Mad Season]
    #23812625 - 11/08/16 12:33 PM (7 years, 3 months ago)

Post deleted by PNW_Shroomer

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InvisibleMad Season
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Re: Contamination Issues....again [Re: PNW_Shroomer]
    #23812645 - 11/08/16 12:39 PM (7 years, 3 months ago)

Larger grains are much harder to sterilize because of the surface area/insulated air pockets. This means that if there's a high number of endospores before your PC cycle, you'll undoubtedly see some leftover CFUs after the cycle.

But considering op is having every jar contaminate on him, this means theres 2 options. Terrible prep, or a contaminated syringe. My money's on the latter, and by cleaning up cultures on agar, you can not only ensure your sterile techniques are flawless, you'll also have 100% clean inoculant, even if your syringe is almost pure bacteria


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.

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OfflineMagicFingers
Stranger


Registered: 04/30/15
Posts: 214
Last seen: 7 years, 5 days
Re: Contamination Issues....again [Re: Mad Season]
    #23814409 - 11/08/16 10:19 PM (7 years, 3 months ago)

In all honesty I wanted to work with rye berries but haven't found a whole food store local that doesn't charge an arm and a leg. WBS is great for spawn however its a pain to work with. It can get messy and a pain to clean up. Either way, the issue is still present likely due to an issue with my PC method. and/or inoculation.

The reason I am thinking that is because I have had a dirty spore syringe and you could see the contaminants that grew in the jar was directly in the path of the inoculation port. You could see Mycelium on the top and bottom and little blackish green dots all the way in-between. You could see the shape of where the water came down. All tests in the future showed the solution was contaminated right off the bat.


I know my savior is going to be Agar. I really have no good excuse for not doing it already. There are endless reasons to go to agar. #1 being it is impossible for anyone to promise zero contaminants in a syringe. #2 you can isolate genetics to find strong growing mycelium. It would be neat to see a culture strong enough to colonize spawn within 10 days! I seriously just need to bite the bullet and order the items needed to do agar. I know pasty had a nice easy tek.



Another quick question. I have noticed some other jars of mine have something I would call Mucky feet? The bottom of the jars show some wetness. Likely due to gravity pulling down the water content to the bottom of the jar. I'm not 100% sure if it is mucky looking grain or actual bacterial growth. What do you experts think? I cannot exclude the option that I didn't clean the grain enough. I also used a little bit of gypsum...  Thoughts?



--------------------
Live the Life you Love!
-MagicFingers:mushroom2:

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OfflinePNW_Shroomer
Cultivator

Registered: 11/08/16
Posts: 42
Loc: Pacific Northwest
Last seen: 7 years, 3 months
Re: Contamination Issues....again *DELETED* [Re: MagicFingers]
    #23815166 - 11/09/16 04:41 AM (7 years, 3 months ago)

Post deleted by PNW_Shroomer

Reason for deletion: ...


Edited by PNW_Shroomer (11/09/16 04:45 AM)

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OfflineMagicFingers
Stranger


Registered: 04/30/15
Posts: 214
Last seen: 7 years, 5 days
Re: Contamination Issues....again [Re: PNW_Shroomer]
    #23815506 - 11/09/16 08:27 AM (7 years, 3 months ago)

Just purchased some agar agar bars. I'm going to follow pasty's agar tek and do this shit right! No More Excuses! :grin:


For reference:  https://www.shroomery.org/forums/showflat.php/Number/19208976/fpart/1/vc/1


--------------------
Live the Life you Love!
-MagicFingers:mushroom2:

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OfflineMagicFingers
Stranger


Registered: 04/30/15
Posts: 214
Last seen: 7 years, 5 days
Re: Contamination Issues....again [Re: MagicFingers]
    #23830770 - 11/13/16 10:40 PM (7 years, 3 months ago)

I just wanted to follow up on this for a couple reasons. The main reason is for anyone that stumbles on this post and would like to see how these turned out in the end.

As expected 4 of the 5 jars are completely contaminated with some kind of white cotton like mold. If one is not familiar with this you may think it looks like mycelium. Usually in the past I will shake a jar to see if it recovers. If it recovers it will likely be ok. However, in some instances I was 99% sure it looked ok. But it was not. Which brings me to the second reason I am posting.

The 5th jar in my opinion actually looks... ok. I would still expect the mycelium to recover faster. But, I am not sure I am seeing any major issues. In the past I wouldn't even consider a G2G with a subject like this. I would just spawn it if it recovers. But mycelium sometimes looks like fuzzy mold. But will usually colonize the spawn quickly. Like, within days.

Again, these were all done at the same time with the same LC.

But I would like to get the opinion from THE EXPERTS! :wink:


Here are the 4 bar jars. You can see they are almost 100% colonized with the nasty nast! After I post this I'll be going outside and disposing of these and leaving the jars soak in some cleaner outside. (which also makes me question how others clean their nasty jars? What I do is hand wash with some heavy duty cleaner and then put them in the dishwasher.)




Here is the ....ok  Looking jar. But what do you think?


--------------------
Live the Life you Love!
-MagicFingers:mushroom2:

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