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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27016137 - 11/01/20 05:54 PM (3 years, 2 months ago) |
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Quote:
Mateah said: Practically every type of bacteria that's in our home/substrates divide every 20-30min on average in good conditions. They look wimpy as hell on agar but in liquids they talk a whole lot of smack, really fast as well and they back that shit up. Reinforcements are always right around the corner and you're already fucked 
There's an unfathomable amount of bacteria out there. Even in beer there's a few dozen common contaminants let alone rare ones. And beer is a environment with a pH that's rough, hop acids that are antibacterial to most bacteria, co2, no air, pressure, yeast has some defenses, and low leftover nutrition. Some bacteria may only grow to a few hundred cells per mL which is hard to detect with all but PCR
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sonoramo
Contaminant



Registered: 02/27/19
Posts: 851
Loc: California, baby!
Last seen: 2 days, 4 hours
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: bodhisatta]
#27042223 - 11/16/20 01:14 PM (3 years, 2 months ago) |
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Quote:
bodhisatta said: There's an unfathomable amount of bacteria out there. ... Some bacteria may only grow to a few hundred cells per mL which is hard to detect with all but PCR
So,... looking for bacteria in the LC with a microscope isn't likely to help?
Would it be a worthwhile "community project" to collect contaminant reports from LC rejects? It would be great if we knew what contaminants really are most common across different locations. For example, I dumped one LC yesterday because a test plate grew trichoderma. It took 3 days to sporulate in the petri, alongside the mycelium.
I've never found bacteria in my LC test plates, but maybe I just don't know how to look for them. When I make a test plate, I try to catch just clumps of growth after spinning the stir bar as fast and hard as I can make it go. Is that the way to get a representative sample? How large a sample do you inject onto the agar? Are there dyes or other ways to make bacteria more visible in the test plate? Maybe I should be testing pH as well?
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: sonoramo]
#27042539 - 11/16/20 04:58 PM (3 years, 2 months ago) |
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Quote:
sonoramo said: Would it be a worthwhile "community project" to collect contaminant reports from LC rejects?
That would be a very worthwhile project in my opinion! I encourage everyone to post their LC's in this thread so we have a variety of different LC 'situations' documented to easier map all the visual indications!
Quote:
I've never found bacteria in my LC test plates, but maybe I just don't know how to look for them.
That's because the standard agar recipe of 2% agar isn't very suitable for detecting bacterial growth. The vast majority of bacteria requires a liquid environment to grow and unless the bacteria is very motile they would in addition need moving water to be able to spread their colonies since most baciera are non motile. That's why microbiologists use mostly liquid and sometimes semi-solid media's like motility agar for propagation. Even tho bacteria have the ability to deal with differences in osmotic pressure, some microbes are better at this some are do not handle lower osmolarity all that well. All in all bacteria as an organism no matter of strain all require extremely high water activity rates compared to for example myc or mold, so the wetter the environment the more suitable it is for bacterial growth. Hope that helps!
BTW you have any LC projects going on right now?
-------------------- Cakes inside Water Tub
Edited by Mateja (11/16/20 05:01 PM)
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sonoramo
Contaminant



Registered: 02/27/19
Posts: 851
Loc: California, baby!
Last seen: 2 days, 4 hours
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27042575 - 11/16/20 05:21 PM (3 years, 2 months ago) |
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Quote:
Mateah said: That's because the standard agar recipe of 2% agar isn't very suitable for detecting bacterial growth. The vast majority of bacteria requires a liquid environment to grow and unless the bacteria is very motile they would in addition need moving water to be able to spread their colonies since most baciera are non motile. That's why microbiologists use mostly liquid and sometimes semi-solid media's like motility agar for propagation. Even tho bacteria have the ability to deal with differences in osmotic pressure, some microbes are better at this some are do not handle lower osmolarity all that well. All in all bacteria as an organism no matter of strain all require extremely high water activity rates compared to for example myc or mold, so the wetter the environment the more suitable it is for bacterial growth. Hope that helps!
Thanks. That is helpful. Most of my experience with bacterial colonies is on food in the refrigerator. Eventually they get big enough to see (and smell!) easily. I'll take a "bottom line" that maybe it isn't so bad if the extra liquid from inoculating a test plate softens the agar.
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sonoramo
Contaminant



Registered: 02/27/19
Posts: 851
Loc: California, baby!
Last seen: 2 days, 4 hours
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27042587 - 11/16/20 05:30 PM (3 years, 2 months ago) |
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Quote:
BTW you have any LC projects going on right now?
I tried to answer by PM so the rest of the community doesn't get sidetracked, but your account doesn't accept PMs. So,...
Yes:
- GT's (now fruiting and so done with it, but saving LC in the refrigerator)
- Unknown cubes from deep in my spore-print collection but they are old friends

- Cambodians, almost ready to inoculate
- Quilla, very ready, will probably inoculate this week
- Oyster culture that's previously "worked" from agar and cardboard, now ready with LC
- Tarragon oyster from commercial syringe, trying to expand before inoculating
In all cases (other than the commercial syringe) it's taken some effort to go from spores to agar to LC, but I seem to get better at it as I go.
My current interest is growing oysters in larger quantities. I just can't consume cubes as fast as I could grow them now that I have a working LC (or four...).
I'm using 16ga dispensing (blunt) needles to sample from agar culture into squat 4oz mason jars. If a culture seems to be clean, then move up to a pint mason jar. Finally, inoculate oats with 20ml of LC, and I've seen full colonization in about 1.5 to 2 weeks. In all cases, lids with SHIP and Tyvek filters. Newer lids I've used two layers of Tyvek in case the broth splashes.
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: sonoramo]
#27042630 - 11/16/20 05:55 PM (3 years, 2 months ago) |
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My account doesn't accept PM's? You mean the automatic message said to "ask cultivation related questions in the public forum"? I guess it's aight to send me links or pics if you want but in general it's much more productive for everyone if we conduct research here instead of PM's 
Do you have pics of any of the LC's you have going right now?
-------------------- Cakes inside Water Tub
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sonoramo
Contaminant



Registered: 02/27/19
Posts: 851
Loc: California, baby!
Last seen: 2 days, 4 hours
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27042744 - 11/16/20 07:05 PM (3 years, 2 months ago) |
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Quote:
Mateah said: My account doesn't accept PM's? You mean the automatic message said to "ask cultivation related questions in the public forum"?...
More like a message that said "this user doesn't accept PM's."
Quote:
Do you have pics of any of the LC's you have going right now?
OK, here are a few samples of what's cooking.
Mexican strain from Quilla (thanks, CaptainFuture). This has pretty much filled the LC, and if I don't withdraw some of it soon it will form floating pads on the surface.

A well-used GT culture (from a local friend), used to inoculate 7 jars so far. I'm done with this because it's been so prodigious. It has formed substantial mycelial mats on the surface.

Commerical tarragon oyster culture from a syringe, expanding it to a pint jar. It's only been there for a few days. Much of what you see is sediment from the MEA powder.
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maxmush
Always learning...

Registered: 06/13/20
Posts: 440
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: sonoramo]
#27042807 - 11/16/20 07:49 PM (3 years, 2 months ago) |
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Following this.
I have had beautiful looking, clear LC's with strong myc growth that when put to agar are contaminated. I have been lucky with LC's with over 95% success rate, but this has happened more than once. My SOP now includes a mandatory agar test before using.
I would love a visual "test", but i am not sure its possible and/or accurate. There are a multitude of bacteria and molds and each would elicit a different visual outcome IMO.
On a side note: I notice nutrient LC's seem to have a higher affinity to contamination. When i use a simple 4% karo solution, I usually have no issues.
-------------------- Disclaimer: all information presented is intended for educational purposes only. All photos are only representations and not directly from the user.
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MLPismyOPSEC
That One Ponyfucker


Registered: 11/13/18
Posts: 884
Loc: Equestria? Mordor? Wester...
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: maxmush]
#27044391 - 11/17/20 06:43 PM (3 years, 2 months ago) |
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Mostly posting to subscribe, but for some reason my LC broths always come out of the PC as a really nice golden color. Munton's LME like most everyone uses
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Lemgrub


Registered: 04/02/18
Posts: 589
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: MLPismyOPSEC]
#27044514 - 11/17/20 08:00 PM (3 years, 2 months ago) |
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These are both inner tissue LCs. Left one was made 11/14 the right was 11/13. I think I can tell which will be no good at this point. Also think I'll tone down the sugar content next time. They're 4% honey.
-------------------- We getting that pirate bay, alien shishkebab cordycep money

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mushhead
Potato Devourer



Registered: 08/22/14
Posts: 2,215
Loc: Dimension X-124
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Lemgrub]
#27044547 - 11/17/20 08:30 PM (3 years, 2 months ago) |
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   Right is B+ left is APE. LME heat dissolved into distilled water with a 45min PC cycle. Some sediment but no other microbes save mycelia in here.
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D3_Myc
Weeb Trash



Registered: 05/06/18
Posts: 4,399
Loc: Year Zero
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: mushhead]
#27046036 - 11/18/20 06:02 PM (3 years, 2 months ago) |
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Following, will post some pics in a bit
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BrownBear
Warrior-Traveler



Registered: 06/05/20
Posts: 1,539
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: D3_Myc]
#27046219 - 11/18/20 07:19 PM (3 years, 2 months ago) |
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How would you rate these lc's?
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enteogenio
Stranger
Registered: 10/31/20
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: BrownBear]
#27046265 - 11/18/20 07:40 PM (3 years, 2 months ago) |
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What do you think:

*MS Srynge: GT *Water/Honey (500ml/4ml PC:30min/15psi) *Date: 19/10/20
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coversall
إِنْ شَاءَ ٱللَهُ



Registered: 06/06/20
Posts: 2,749
Loc: संसार
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: enteogenio]
#27049841 - 11/21/20 03:47 AM (3 years, 2 months ago) |
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0.4% LME, PC'd for ~20 mins at 15 psi, noc'd with a pan cyan agar wedge.

0.4% LME, PC'd for ~40 mins at 15 psi, noc'd with a pan cyan needle poke biopsy from a pan cyan agar plate.
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verum subsequentis
seeker of truth



Registered: 03/22/16
Posts: 8,732
Last seen: 1 year, 7 months
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: coversall]
#27050583 - 11/21/20 02:59 PM (3 years, 2 months ago) |
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Sexy
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coversall
إِنْ شَاءَ ٱللَهُ



Registered: 06/06/20
Posts: 2,749
Loc: संसार
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: verum subsequentis]
#27051448 - 11/22/20 02:04 AM (3 years, 2 months ago) |
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Forgive me, but I need to check, is that a sarcastic sexy? Or is the LC looking ok? I gather it's near impossible to tell whether an LC is clean just by looking at it?
I'm going to yolo one of the smaller batches today.
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D3_Myc
Weeb Trash



Registered: 05/06/18
Posts: 4,399
Loc: Year Zero
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: coversall]
#27051693 - 11/22/20 08:24 AM (3 years, 2 months ago) |
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Img dump of the cultures I have going atm. One at rest and one swirled

The tat spun for a few days before I let it settle. The dark spots are the individual tissue pieces
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coversall
إِنْ شَاءَ ٱللَهُ



Registered: 06/06/20
Posts: 2,749
Loc: संसार
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: D3_Myc]
#27051725 - 11/22/20 08:36 AM (3 years, 2 months ago) |
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Are these clean/tested LCs?
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D3_Myc
Weeb Trash



Registered: 05/06/18
Posts: 4,399
Loc: Year Zero
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: coversall]
#27051742 - 11/22/20 08:49 AM (3 years, 2 months ago) |
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No, they are “I don’t even know what I’m looking at cultures and could use some interjections”
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