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OfflineRrrr266
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Honey tek liquid culture with lots of dust-like particles
    #27281726 - 04/25/21 04:17 PM (3 years, 8 months ago)

I am trying to grow liquid culture from spores. The water was always muddy since I used a poorly filtered honey bought straight from the producer. After inoculation tons of tiny particles appeared on day 2. It is 9 days old now. Could it be contaminated?


Video https://streamable.com/j680rb

I also started this jar 5 days ago because the first one looks suspicious. I used the same spores but a different honey bought from a supermarket. It is crystal clear and mycelium looks the same as the first one.


Edited by Rrrr266 (04/25/21 04:20 PM)

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Offlineverum subsequentis
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Re: Honey tek liquid culture with lots of dust-like particles [Re: Rrrr266]
    #27281764 - 04/25/21 04:57 PM (3 years, 8 months ago)

"Could it be contaminated?"

Yes. Of course. Spores to LC is generally a risky game because spores are generally not perfectly clean. It can work though.

From what is presently growing, I see nothing to lead me to believe it's contaminated. Post pics as it grows out and I'll gladly let you know what I think.

PS: I'm going to advise you to test it on agar before sploogin into a bunch of grain jars.

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OfflineRrrr266
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Re: Honey tek liquid culture with lots of dust-like particles [Re: verum subsequentis]
    #27281810 - 04/25/21 05:53 PM (3 years, 8 months ago)

Thanks a lot. Will post more pics as they grow.

I should have started with agar but liquid culture seemed very simple at first glance, turned out I was wrong. Definitely gonna test them on agar before going all in.

Edited by Rrrr266 (04/25/21 05:54 PM)

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Re: Honey tek liquid culture with lots of dust-like particles [Re: Rrrr266]
    #27281950 - 04/25/21 08:41 PM (3 years, 8 months ago)

Looks like Staphylococcus bacteria first pic. Video has a biofilm on top? 3rd pic Bacillus? Shake it if bubbles form on top and chunks flow its aBacillus. Just noticed you used spores to lc. Do agar, 1 spec is all you need to start from multipore to "single". Be able to isolate agar to agar to be sure you're clean. Easier to tell if the problem is your environment and method of sterilaztion.

The jar doesn't look like its meant for canning either, I bet you're using no sealant liner or foam liner. Jars like those which I'm currently using got contamed until I made my own sealant on the jar itself with high temp silicon caulking and parafilm. Always leave 1 jar untouched so you know if the problem is with the jars itself, caramilization of the protien and unfermented sugars or your method.

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OfflineRrrr266
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Re: Honey tek liquid culture with lots of dust-like particles [Re: PieHole]
    #27283248 - 04/26/21 10:15 PM (3 years, 8 months ago)

Quote:


Looks like Staphylococcus bacteria first pic. Video has a biofilm on top? 3rd pic Bacillus? Shake it if bubbles form on top and chunks flow its aBacillus. Just noticed you used spores to lc. Do agar, 1 spec is all you need to start from multipore to "single". Be able to isolate agar to agar to be sure you're clean. Easier to tell if the problem is your environment and method of sterilaztion.




The first one forms bubbles and chunks float on the surfacefor an hour or so  when I shake it. It doesn't happen in the second jar tho. I will test both and the spores on agar to make sure what is going on. Waiting for delivery.

Quote:


The jar doesn't look like its meant for canning either, I bet you're using no sealant liner or foam liner. Jars like those which I'm currently using got contamed until I made my own sealant on the jar itself with high temp silicon caulking and parafilm. Always leave 1 jar untouched so you know if the problem is with the jars itself, caramilization of the protien and unfermented sugars or your method.




They are pretty good. The lids have an elastic layer on the inner side sealing the rims perfectly.

Actually I let the second jar sit for three days before inoculating. Couldn't think of making an extra jar for control lol, will definitely do it next time. Thank you.

Edited by Rrrr266 (04/26/21 10:22 PM)

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Offlineverum subsequentis
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Re: Honey tek liquid culture with lots of dust-like particles [Re: Rrrr266]
    #27283256 - 04/26/21 10:26 PM (3 years, 8 months ago)

I wouldn't bother with the control jar. I'd ditch honey and use malt extract. Doesn't really matter what type of jar you use if it's got a proper filter for breathing and you wrap the lip with glad or parafilm.

I've PCed Malt LC for hours without "caramelization" problems.

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OfflineRrrr266
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Re: Honey tek liquid culture with lots of dust-like particles [Re: verum subsequentis]
    #27299876 - 05/08/21 08:47 AM (3 years, 8 months ago)

I transfered all to agar and finally have some growth identifiable enough(i guess) at day 5. Sorry abut the flash, it is hard to photograph the dishes with lids on.


Spore syringe to agar.


Liquid culture, the muddy one.


Liquid culture, the clear one. There is a single white blob on the top side that looks different to the rest.

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Offlineverum subsequentis
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Re: Honey tek liquid culture with lots of dust-like particles [Re: Rrrr266]
    #27300904 - 05/08/21 10:48 PM (3 years, 8 months ago)

The blob is the only concerning thing i see. I pic tomorrow or the next day and I'll let you know how I feel.

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OfflineRrrr266
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Re: Honey tek liquid culture with lots of dust-like particles [Re: verum subsequentis]
    #27301660 - 05/09/21 01:49 PM (3 years, 8 months ago)




It doubled in size in approx. 30 hours. Gotta test that jar again to make sure if that thing was an airborne contaminant.

The dirty jar surprisingly turned out to be viable for use. Thanks a lot for the help.

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