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Mateja


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LC Testing + Grow Log 1
#26987240 - 10/15/20 02:50 PM (3 years, 4 months ago) |
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I'll try to update this grow log frequently with pics of different LC's and pics of the colonization process of grain and cakes and finally I'll be posting pics of the fruiting results to see if there can be established a clear connection between successful grows and the 'clean look' appearance of the LC broths used.
I started off with this broth made from 0.1% LME and inoculated with an agar wedge on Oct 1st. I will call this the 'Master LC' and here's the looks of it as it was finishing on Oct 10th.

On October 12th this Master was aspired into syringes and used to inoculate 12 new broths via Ship's with 1ml each. (except one qt har that was injected with 5ml) I used nine 250ml jars containing 100ml broth and three qt jars containing 300ml broth. These new broths were made of 0.05% LME and this is of the qt jars 48h after being inoculated with 1ml Master. Before and a after a swirl.
 
On Oct 12th 9 rye jars we're also each one inoculated with 5ml through the SHIP. This is the look of a 72h recovery in 3 of the jars that weren't shook upon inoculation.
  
For this experiment I'm incubating both LC's and jars at around 28C (82f). The reson for this is that I want as prolific growth of potential nasties as possible to easier/sooner be able to detect their presence. Idk what anyone else thinks of this approach I'm open to hearing opinions about perceived pros/cons but for now this is how I'm going about it.
-------------------- Cakes inside Water Tub
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Forrester
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Re: LC Testing + Grow Log [Re: Mateja]
#26987245 - 10/15/20 02:52 PM (3 years, 4 months ago) |
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-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Josex
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Re: LC Testing + Grow Log [Re: Forrester]
#26987279 - 10/15/20 03:14 PM (3 years, 4 months ago) |
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That's a nice recovery on grain man. I'll be monitoring this shit!
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verum subsequentis
seeker of truth



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Re: LC Testing + Grow Log [Re: Josex]
#26987301 - 10/15/20 03:33 PM (3 years, 4 months ago) |
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Mateja


Registered: 07/14/16
Posts: 7,948
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Day 5 offers some interesting observations. I notice turbidity in some of the jars and to varying degrees. Upon inspecting these two jars I observed some turbidity in both, and IMO broth on the left seems slightly more turbid.

Here I compare both of those LC's to a sterilized broth that wasn't inoculated and that doesnt display any turbidity.
 
Here is another couple of LC's that appear to show the same thing, namely the broth on the left being slightly more turbid.

And here they are compared to the sterile broth
 
Here is a broth in which I couldn't notice any turbidity. Pics of that LC before and after a swirl.
 
And here is that LC compared to the sterile broth

The coming days will reveal more clearly what's going on with these LC's and I believe that when I start testing these on agar and grain in a few days it will both be easier to make predictions about the outcome as well as results ending in more definite conclusions. (Or not lol)
-------------------- Cakes inside Water Tub
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Forrester
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Re: LC Testing + Grow Log [Re: Mateja]
#26989532 - 10/17/20 04:05 AM (3 years, 3 months ago) |
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So just to make sure I understand correctly what's going on here...
The (inoculated) jars that you're showing next to each other where one is more clear than the other - those were all inoculated the same and there's no (known) difference other than their appearance at this time, correct?
And you're eventually going to run them across agar to see if there's any contamination showing?
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Mateja


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Re: LC Testing + Grow Log [Re: Forrester]
#26989580 - 10/17/20 04:56 AM (3 years, 3 months ago) |
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Quote:
Forrester said: So just to make sure I understand correctly what's going on here... The (inoculated) jars that you're showing next to each other where one is more clear than the other - those were all inoculated the same and there's no (known) difference other than their appearance at this time, correct?
All 12 broths are the same 0.05% LME broth. 9 of these are 250ml jars and 3 of them are 'qt' jars. All 9 of the 250ml jars and 2 of the qt jars were inoculated with 1ml of Master except for one of the qt jars which was inoculated with 5ml of Master to see how growth rate would be affected. After only a couple of days of incubation there was no longer any noticeable difference in myc mass between that one qt jar that was inoculated with 5ml and the other two qt jars that were injected with 1ml. The 250ml jars are these with the flat, angled, segmented body

And the qt jars in the pics are the ones with a round body (two of the three qt jars are identical with the one uninoculated jar (the sterile broth) these jars have the writing 'mason' on them) and the one 'odd' qt jar is identical to the grain qt jars in these pics. I hope this provides clarity!
So to answer your question fully, in this one here comparison pic that I posted earlier depicts one of the qt jar LC's that was inoculated with 5ml and the other one that was inoculated with 1ml. This is the only variable that breaks the consistency of the parameters but for the purpose of this experiment this variable shouldn't really affect the assessment of this experiment since we're mainly focusing on identifying turbidity, the varience of turbidity between LC's and comparison to the sterile broth which acts as the 'control' jar in this case. These are the two qt jars mentioned that were inoculated with 1ml and 5ml tho from this pic I don't remember which is which, but that shouldn't matter.

Quote:
And you're eventually going to run them across agar to see if there's any contamination showing?
Yes, I will run tests on two types of agar, LME and GWA as well as on rye qt jars and 1/2 pint Oatmeal-Coir cakes.
-------------------- Cakes inside Water Tub
Edited by Mateja (10/17/20 05:01 AM)
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Forrester
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Re: LC Testing + Grow Log [Re: Mateja]
#26989582 - 10/17/20 04:59 AM (3 years, 3 months ago) |
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Ok cool that clears it up a bit. I was mostly just making sure there was no difference in method that made the more and less clear jars
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Mateja


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Re: LC Testing + Grow Log [Re: Forrester] 1
#26989591 - 10/17/20 05:08 AM (3 years, 3 months ago) |
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Well actually not all of the jars were inoculated with the same method. All of the qt jars and 6 of the 250ml jars were inoculated through the SHIP and 3 of the 250ml jars were inoculated by lifting the lid and injecting through the opening. I also purposefully inoculated some jars more sloppy than others. All of this was done to try to create some variance that would hopefully result in some jars getting clean inoculation and some jars getting contaminated upon inoculation so that I'm actually able to observe contaminating broths and compare them to clean ones
-------------------- Cakes inside Water Tub
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Forrester
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Re: LC Testing + Grow Log [Re: Mateja]
#26989594 - 10/17/20 05:12 AM (3 years, 3 months ago) |
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Ah, ok, now that makes a difference. That's what I was looking for. Will be interested to see the results!
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Mateja


Registered: 07/14/16
Posts: 7,948
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Re: LC Testing + Grow Log [Re: Forrester]
#26989616 - 10/17/20 05:58 AM (3 years, 3 months ago) |
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Btw thanks for asking for clarifications, that helps this thread to be expressed more clearly, and also helps me to remember to include all the data from all the different tests going on in this experiment  So with that said here comes an update on the colonizing rye jars that were inoculated with the Master as well, and to further clarify all 9 jars were inoculated through the ship and 5 of them were shaken upon inoculation and 4 weren't. This also as an attempt to determine how possible contamination can be detected on grain that was shook upon inoculation vs grain that wasnt.
So on day 5 this is the look of two of the rye jars that weren't shaken upon inoculation.
 
This is the only one out of the 5 'shaken upon inoculation' jars that shows recovery at this moment.

And these are the three Oatmeal-Coir cakes that were inoculated with 5ml Master each with the standard PFTek inoculation procedure. They were inoculated on Oct 14th, 2 days after the broths and rye jars were inoculated. So this is how recovery looks on cakes on day 3.
 
-------------------- Cakes inside Water Tub
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eLeSDenes
Mycelium Expander




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Re: LC Testing + Grow Log [Re: Mateja]
#26989665 - 10/17/20 07:09 AM (3 years, 3 months ago) |
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what is the point of master LC? why not just make a new LC from wedges? I think it carries way more risk and does not take less time than dropping a wedge.
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Mateja


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Re: LC Testing + Grow Log [Re: eLeSDenes]
#26989697 - 10/17/20 07:51 AM (3 years, 3 months ago) |
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Quote:
eLeSDenes said: what is the point of?
Basically is just an experiment testing if/how visual cues can be interpreted when it comes to determining quality of colonizing broths and substrates. In other words looking to see if there's a sure way to tell the quality of the sterile processes at all times and especially during production of inoculate. But to adress your question yes I could have used agar cultures as well to inoculate the broths, it wouldn't matter so much in this case, tho I probably will continue this grow log indefinitely for now and will make many more broths which will even be inoculated with specific bacterial colonies that I have saved on plates. This will be an unusual grow log I guess with no clear plan of path, the insentive at least is to possibly debunk or confirm some widely debated topics. Like visual inspection of a LC for example.
-------------------- Cakes inside Water Tub
Edited by Mateja (10/17/20 07:53 AM)
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BrownBear
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Re: LC Testing + Grow Log [Re: eLeSDenes]
#26989706 - 10/17/20 08:02 AM (3 years, 3 months ago) |
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I am really digging this thread as I have always been a LC enthusiast. The grain test on shake vs no shake is really intersting also. I am excited to see how it all progresses.
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eLeSDenes
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Re: LC Testing + Grow Log [Re: Mateja]
#26989735 - 10/17/20 08:37 AM (3 years, 3 months ago) |
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Quote:
Mateah said:
Quote:
eLeSDenes said: what is the point of?
Basically is just an experiment testing if/how visual cues can be interpreted when it comes to determining quality of colonizing broths and substrates. In other words looking to see if there's a sure way to tell the quality of the sterile processes at all times and especially during production of inoculate. But to adress your question yes I could have used agar cultures as well to inoculate the broths, it wouldn't matter so much in this case, tho I probably will continue this grow log indefinitely for now and will make many more broths which will even be inoculated with specific bacterial colonies that I have saved on plates. This will be an unusual grow log I guess with no clear plan of path, the insentive at least is to possibly debunk or confirm some widely debated topics. Like visual inspection of a LC for example.
Ahh cool man interesting. I am also using 0.1% LME to get really clear looking LC. I think clearness of LC is not an indication of being clean ( there is no bacteria in it). I had lots of super clean looking LC which somehow seemed to stall on grains after shake.
I am happy that someone else as well investigate this topic as LC's are super easy and fast. Interested to see your results and conclusions.
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verum subsequentis
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Re: LC Testing + Grow Log [Re: eLeSDenes]
#26989956 - 10/17/20 11:30 AM (3 years, 3 months ago) |
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I've been out of the game for a quick minute but I'm also really into Liquids. Me rikey this thread. It's maturing rather nicely. When i get back up and rainbow farming i'd love to do more exploring and documenting Liquids.
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A.k.a
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That rye growth is awesome.
I’m curious how the shaken vs not jars end up. When I shake mine up it seems to take forever to see recovery but then the whole jar goes off. By the looks of it the not shaken ones here are so far ahead they’ll definitely finish before the others.
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LAGM2020     
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Re: LC Testing + Grow Log [Re: A.k.a] 1
#26990010 - 10/17/20 12:11 PM (3 years, 3 months ago) |
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I've found that shaking the grain jars nocced with LC really slows down the bounce back. I theorize that this is due to it being very vulnerable because it hasn't colonized anything that can protect it. I believe that you can shake grain and see it bounce back in a day or two because the myc INSIDE the grain remains undamaged and therefor bounces back. Where as the myc from the LC just gets beat to shit and has nowhere to hide.
To test this I did a batch in which i nocced up a bunch of masters with the same LC. Some i shook vigorously, some i rolled gently, and some i didn't shake at all. The shaken jars chilled with no growth for quite some time while the gently rolled jars exploded with growth all over in a few days. The jars left alone did what LC jars always do when everything is done right. The rolled and non shaken jars (which got shaken once the myc had colonized a good portion of the jar) colonized in about the same time.
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A.k.a
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I definitely agree about the internal colonization thing. Next time I’m gonna let it sit for 2-3 days and then shake it. There’s gotta be a sweet spot where the myc gets established enough that shaking will speed things up.
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LAGM2020     
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Forrester
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Re: LC Testing + Grow Log [Re: A.k.a]
#26990117 - 10/17/20 01:47 PM (3 years, 3 months ago) |
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That is very interesting about the shaking w/LC. I always shook my jars, looks like I'll have a new way to do it now
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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