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 Mateja
 
Registered: 07/14/16
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 LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth   6
#27007467 - 10/28/20 12:35 AM (4 years, 1 month ago) |
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This experiment will demonstrate that bacteria will cause turbidity in a LC and that turbidity is easily recognized.
5 nutrient rich liquid mediums were used for this test. The nutrient density is purposefully kept to a minimum so that the liquid medium will be crystal clear and able to effectively contrast turbidity. Three of these broths contain 0.05% malt extract by weight and the other two were made from diluted rye grain water which was then further diluted by mixing with 19 parts water.
Bacteria and mold colonies used for this test were gathered from these plates
 
1 ME broth was inoculated with a grey/opaque bacterial colony, 1 ME broth was inoculated with a yellow bacterial colony and 1 ME broth was inoculated with a mold colony. 1 GW broth was inoculated with a wedge containing both cube myc and the grey/opaque bacteria and the other GW broth was inoculated with a wedge containing both mold and cube myc. These broths were incubated at 25C (77F) and swirled once a day.
The 5 broths shortly after inoculation:
To inspect for turbidity an object should be placed right behind the LC jar along with a back light that will illuminate both broth and the object. I chose to use a small spore solution vial but any kind of object can be used as long as it has distinct patterns/lines that can be easily observed through the crystal clear LC jar.
Example:
After only 8h of incubation we can already observe noticeable turbidity in LC's that were inoculated with bacteria and the spore vial can not be seen through all the light diffusion inside the liquid media. The light scatters in all directions as it reflects off of countless bacterial cells. LC's containing only mold and/or cube myc are still transparent and the spore vial can be observed through the broth.
Results:
 
 
The LC's in those pics above were inoculated with bacterial agar wedges and regardless how tiny the bacterial colonies were they still contained a huge number of cells which contributed to a very fast colonization of the broth. For the next test I will use just a fraction of that and let's see what will happen. These next jars were both first inoculated with 10ml of mold liquid culture each using a syringe and needle. One of these mold LC's was then inoculated with one drop of bacterial solution and the other mold LC was inoculated by gently touching the broth surface with the needle tip of that same needle.
The broth that was inoculated with 1 drop of bacterial solution went turbid within a few hours and the mold colony appeared to stop growing right away. The other (needle tip) broth offered the mold colony a chance to recover briefly but within 24h it also went turbid and no more growth could be observed in the mold colony.
Results:
 
In the pics below I didn't even use a background object, only light, and the bacterial LC can easily be differentiated from the clean LC and from the control jar which hadn't been inoculated.
Results:
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Cakes inside Water Tub
Edited by Mateja (03/10/22 01:00 AM)
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Baba Yaga
Psychedelic Minion
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 Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27007489 - 10/28/20 01:00 AM (4 years, 1 month ago) |
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Thanks for doing this.
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Mateja
Registered: 07/14/16
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Baba Yaga] 1
#27007624 - 10/28/20 04:38 AM (4 years, 1 month ago) |
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And it seems that bacteria will fuck up a liquid broth in a matter of hours 
I placed a spore solution vial inbetween the light and the LC jar as to be able to gauge the anticipated turbidity.
But I guess I should have takes pics every 30min or so lol these photos were taken 8h efter inoculation and that's all folks for this experiment 
Soooo... If your LC hasn't gone turbid in the last few hours I'd say you can be fairly sure there's not bacterial activity in there. 
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Cakes inside Water Tub
Edited by Mateja (10/28/20 04:39 AM)
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bodhisatta  
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja] 3
#27007627 - 10/28/20 04:42 AM (4 years, 1 month ago) |
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Some bacteria will cause turbidity some wont. Some bacteria form biofilms because of their secretions. Those make a broth way more turbid than say most species of lactobacillus would and some lacto make no turbidity
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Mateja
Registered: 07/14/16
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: bodhisatta]
#27007640 - 10/28/20 04:58 AM (4 years, 1 month ago) |
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I will continue this experiment for the sake of documenting different types of bacteria and mold growth in LC's to learn to decide easier upon future visual inspections. I can understand how some microbes due to secreting make the broth even more turbid. But are you saying that there are types of bacteria that consist of a perfectly transparent cell? And even when those cells are jammed in the broth in astronomical numbers should there really still be no visible turbidity?
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Cakes inside Water Tub
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dfwerydfhg
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27007813 - 10/28/20 08:01 AM (4 years, 1 month ago) |
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Good stuff. Now who has a spectrophotometer?
Would be interesting to see what happens when you add bacteria, either via agar like you did or a drop of your new bacteria broth, into a clean colonized or colonizing LC.
Inoculation size might matter in some situations- I'm guessing your additions were orders of magnitudes larger than the start of a typical contam. More of an issue in the case where myc might otherwise be able to fight it.
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Mateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: dfwerydfhg]
#27008660 - 10/28/20 03:22 PM (4 years, 1 month ago) |
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Quote:
dfwerydfhg said:
Good stuff. Now who has a spectrophotometer?
You have two of them on your face, kind of 
Quote:
Would be interesting to see what happens when you add bacteria, either via agar like you did or a drop of your new bacteria broth, into a clean colonized or colonizing LC.
Yes I will be doing that as well, adding mold and bacterial colonies to clean LC's at various stages to see how growth is affected.
Quote:
Inoculation size might matter in some situations- I'm guessing your additions were orders of magnitudes larger than the start of a typical contam. More of an issue in the case where myc might otherwise be able to fight it.
Yes these bacterial colonies were gigantic in terms of numbers even tho they were tiny specks. Next round i will just dip the tip of the needle or gently swipe a bacterial colony on agar and then just gently touch the surface of the broth with the tip of the needle. Now that I think of it I actually know a couple of ways to dilute bacterial colonies so I end up with just a handful of cells, I might do that as well if dipping the tip of the needle proves too crude as well.
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Cakes inside Water Tub
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 Danger-Dave 1
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27008677 - 10/28/20 03:34 PM (4 years, 1 month ago) |
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Yeah! Thank you for doing this experiment! this needed to be done!
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verum subsequentis
seeker of truth
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Danger-Dave 1]
#27008740 - 10/28/20 04:10 PM (4 years, 1 month ago) |
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Josex
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: verum subsequentis]
#27008754 - 10/28/20 04:21 PM (4 years, 1 month ago) |
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Oh shit dude, was it my penis that gave you that nasty plate? I think I can even see some AIDS there too. Cool experiment tho.
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Mateja
Registered: 07/14/16
Posts: 7,948
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Josex]
#27008813 - 10/28/20 04:55 PM (4 years, 1 month ago) |
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I've seen much worse 
and if I didn't have faith in your work I'd probably put the swabs to more than 2 plates.
I'm pretty sure I found a handful of cube germ spots in this one plate that I transferred, I'll know in a day or two 
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Cakes inside Water Tub
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Josex
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27008834 - 10/28/20 05:11 PM (4 years, 1 month ago) |
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I always assume a swab is dirty though. These fruits were all flopping big dicks bending over with the caps touching the surface, many of them with the gills facing upwards, so some nastiness is to be expected.
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Mateja
Registered: 07/14/16
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Josex]
#27008941 - 10/28/20 06:11 PM (4 years, 1 month ago) |
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At no point did I ever consider that the swabs you sent me were clean lol I just have faith that you're not sending me a catastrophe worth shit.
I also have faith in my ability to get dirty spores to germinate successfully on a grow medium
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Cakes inside Water Tub
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Mateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27015688 - 11/01/20 12:38 PM (4 years, 1 month ago) |
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This time I inoculated two new broths with 10ml of mold each and then I added
one drop of bacterial solution to one of them and I gently dipped the tip on the bacterial needle into the other broth.
The jar that got a drop of bacterial solution immediately colonized the broth within hours and completely stopped the mold in its tracks. The broth that was dabbed with the bacterial needle tip offered the mold a chance to gain some momentum for probably 24h or so but the day after inoculation I already noticed obvious turbidity. That's also why one of the broths has a larger mold colony than the other, as visible in the pics. When it comes to the bacterial activity in these broths I'd say both reached 'stationary phase' within 36h, the onset of turbidity was gradual but extremely fast and hasn't increased since. So far I'm observing maximum cell mass reached within a couple of days in 200-300ml broth.
Here is the 'drop' and the 'needle tip' inoculated on 31st
And just for reference, here are two homogenized mold LC's with crystal clear broths.
One is MEA 0.05% the other is boiled rye broth diluted with 19 parts water.
Lately I've been placing this Pan Cyan vial inbetween the broths and the light source as a means to detect turbidity. I don't have a spectrophotometer so the text on the spore solution vial is my tool for measurements. On the other hand I don't really have much need for precise measurements or estimations using devices since I only need to detect any turbidity whatsoever or the complete lack thereof.
Mold in MEA and rye water
 
The Pan Cyan vial is behind the bacterial LC's as well but the light diffusion inside the solution makes is virtually impossible to see, light scatters all over the place trying to shine through a solution crammed with ("invisible") cells.
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Cakes inside Water Tub
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Josex
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27015704 - 11/01/20 12:50 PM (4 years, 1 month ago) |
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That has been my experience, the few times I got bacteria in an LC it would go to shit fast from just a little needle poke. It got turbid very quickly and the cube myc reacted to the bacteria in a pretty obvious way, it would slow down its growth too. I for one am dying to see this sneaky bacteria that won't cause turbidity in the liquid that all people talk about, the one that only shows it's ugly face on agar, or even an LC that only has 'a little of bacteria' after being colonizing for more than one week. 
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Mateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Josex]
#27015767 - 11/01/20 01:31 PM (4 years, 1 month ago) |
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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
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Cakes inside Water Tub
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tripdawg420
low life with no life
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja] 1
#27015804 - 11/01/20 01:54 PM (4 years, 1 month ago) |
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Josex
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Registered: 11/13/15
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: tripdawg420]
#27015874 - 11/01/20 02:42 PM (4 years, 1 month ago) |
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I would like to see you try inoculating a healthy 100% cube LC with bacteria, that should be interesting too. Now that you're all in with this I'm sure you won't mind. 
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Mateja
Registered: 07/14/16
Posts: 7,948
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Josex]
#27015952 - 11/01/20 03:53 PM (4 years, 1 month ago) |
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Quote:
Josex said:
inoculating a healthy 100% cube LC with bacteria
No sweat man, I was gonna play the villain anyway
Was thinking of squirting moderate/excessive amounts of bacterial solution into a fully or mostly colonized myc LC and then immediately use that to inoculate cakes and grain. Been thinking of doing this for a long time cause I'm curious to see how bacterial gangs go about handling their business on the turfs. I've never (I'm assuming no one has) seen bacteria in action on grain with visible visibility from the eyes only. I'd just pour a 50/50 solution myc/bac on top of rye to see if myc colony will be able to implement their policy and ideology or if they'd all be corrupted resulting in bad guys taking over and ruining the society. And then I'd do the same to cakes. This test will probably have varying results tho depending on the water activity of the grain and cake turf it encounters.
Okej kourrwa I have all the passwords and the keys are activate, I just have to mine some more Mycelium to put inside the warhead together with Bacillus so it's will be weaponize properly kourrwa!
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Cakes inside Water Tub
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verum subsequentis
seeker of truth
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27016056 - 11/01/20 05:00 PM (4 years, 1 month ago) |
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I think it'd be interesting to test an LC on agar and verify it is clean. Then inoculate something. Then spit in it, put it on a stir plate or swirl it and noc something else. Then noc a new jar every day and track the progress.
But aint nobody got time for that.
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