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Amanita virosa
botanist by day


Registered: 12/04/11
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Bench top myco filtration experiment. Complete with control
#18878176 - 09/22/13 09:55 PM (10 years, 4 months ago) |
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So, here the idea is to do test the stropharia for its ability to filter ecoli in a lab environment. I grew the ecoli in lb broth. I then used a sep funnel to slowly drip the broth thru a bag of sawdust and a bag of stropharia. Next, I captures the effluent, and I did serial dilutions from 1 to .0000001 ( very time consuming). Then I plated one microliter from each serial dilution onto agar and counted colonies of bacteria. I will post more pix with results soon.


Edited by Amanita virosa (09/26/13 07:40 AM)
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leschampignons
Biochemistry + Mycology


Registered: 08/30/13
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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18880148 - 09/23/13 11:16 AM (10 years, 4 months ago) |
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cool setup. I would think it would be easier to show this in liquid culture though - liquid+bacteria+myc=nomnomnom. might take a bit longer though.
Curious to see results. The stropharia doesn't mind have liquid dripped through it?
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Jeff
Addict



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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18882137 - 09/23/13 08:17 PM (10 years, 4 months ago) |
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Holy crap bro. You have a hell of a lot more patience than I have. Nice work and waiting the results.
-------------------- Myco-tek
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cubenpete
Aminita good excuse



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Re: Bench top myco filtration experiment. Complete! [Re: Jeff]
#18882165 - 09/23/13 08:26 PM (10 years, 4 months ago) |
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SCIENCE! How very cool..
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Forrester
aspiring sociopath


Registered: 02/05/13
Posts: 9,351
Loc: Northeast USA
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Re: Bench top myco filtration experiment. Complete! [Re: cubenpete]
#18882337 - 09/23/13 09:01 PM (10 years, 4 months ago) |
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Awesome! Just curious, why stropharia? Better than oysters/reishi? (I know nothing of this stuff, but they seem to like to kill bacteria)
-------------------- 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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drake89
Mushroom Magnate



Registered: 06/26/11
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Re: Bench top myco filtration experiment. Complete! [Re: Forrester]
#18882361 - 09/23/13 09:05 PM (10 years, 4 months ago) |
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Quote:
Forrester said:

Awesome! Just curious, why stropharia? Better than oysters/reishi? (I know nothing of this stuff, but they seem to like to kill bacteria)
That's what stamets talks about in mycelium running, where he figured his giant patch was eating e.coli.
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Forrester
aspiring sociopath


Registered: 02/05/13
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Re: Bench top myco filtration experiment. Complete! [Re: drake89]
#18882391 - 09/23/13 09:08 PM (10 years, 4 months ago) |
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Quote:
drake89 said: That's what stamets talks about in mycelium running, where he figured his giant patch was eating e.coli.
Nice, figured there was a reason. Haven't been able to check out the book yet.
-------------------- 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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Amanita virosa
botanist by day


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Re: Bench top myco filtration experiment. Complete! [Re: Forrester]
#18884130 - 09/24/13 09:56 AM (10 years, 4 months ago) |
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Quote:
Forrester said:
Quote:
drake89 said: That's what stamets talks about in mycelium running, where he figured his giant patch was eating e.coli.
Nice, figured there was a reason. Haven't been able to check out the book yet.
Actually I am testing four species. Stropharia p. pulmonarius p. columbines h. Ulmarius (real elm not pseudo) this is a trial run
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generalsherman55
MF BOOGNISH


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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18884500 - 09/24/13 11:23 AM (10 years, 4 months ago) |
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the amount of science that went into this is beyond my reckoning ....
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Amanita virosa
botanist by day


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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18885174 - 09/24/13 02:13 PM (10 years, 4 months ago) |
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Here are the results! Pretty exciting. Each circle origionated from one bacterial colony. The These are the .000001, .00001, and .0001 dilutions. Clearly the stropharia had an negative effect on the number of bacteria. It seems to act as a filter that is better than substrate alone.
Edited by Amanita virosa (09/25/13 10:38 PM)
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liamtheloser
Advanced Idiot

Registered: 06/07/06
Posts: 1,453
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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18885189 - 09/24/13 02:18 PM (10 years, 4 months ago) |
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Where's the control
--------------------
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Amanita virosa
botanist by day


Registered: 12/04/11
Posts: 2,458
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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18885214 - 09/24/13 02:28 PM (10 years, 4 months ago) |
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Good question! Still in incubator when I took these pix.. There was a three hour lag time between when I did these two dilutions and when I did the dilutions of the broth. I had to teach a lab in between. Will post more pix tomorrow but it's looking like the sawdust did act as a filter as well. Just not as good of one as the myc.
Edited by Amanita virosa (09/25/13 10:56 PM)
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liamtheloser
Advanced Idiot

Registered: 06/07/06
Posts: 1,453
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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18885228 - 09/24/13 02:33 PM (10 years, 4 months ago) |
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Cool, I figured 2 bags of sawdust would probably do some pretty good filtering by themselves, was just curious what it would look like.
Thanks!
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drake89
Mushroom Magnate



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Re: Bench top myco filtration experiment. Complete! [Re: liamtheloser]
#18885760 - 09/24/13 04:44 PM (10 years, 4 months ago) |
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looks like ya got some statistical significance there
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pseudotsuga


Registered: 06/29/11
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Re: Bench top myco filtration experiment. Complete! [Re: drake89]
#18885983 - 09/24/13 05:40 PM (10 years, 4 months ago) |
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Amazing work AV! This is one of the most convincing experiments in mycoremediation I've ever seen. Thank your for taking the time to do some solid experimentation.
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liamtheloser
Advanced Idiot

Registered: 06/07/06
Posts: 1,453
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Re: Bench top myco filtration experiment. Complete! [Re: pseudotsuga]
#18890722 - 09/25/13 06:51 PM (10 years, 4 months ago) |
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Where's that control!
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drake89
Mushroom Magnate



Registered: 06/26/11
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Re: Bench top myco filtration experiment. Complete! [Re: liamtheloser]
#18890733 - 09/25/13 06:53 PM (10 years, 4 months ago) |
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I don't know if you're trying to get published, joe, but if you are, the only paper on mycofiltration I could find last year was some kid at evergreen's senior research project. Spread the word.
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Amanita virosa
botanist by day


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Re: Bench top myco filtration experiment. Complete! [Re: liamtheloser]
#18890758 - 09/25/13 06:59 PM (10 years, 4 months ago) |
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Damn son. It's comin'. Forgot my iPad today (camera). Will post tomorrow am after my lab. IQuote:
liamtheloser said: Where's that control!
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RogerRabbit
Bans for Pleasure



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Re: Bench top myco filtration experiment. Complete! [Re: Amanita virosa]
#18891585 - 09/25/13 10:18 PM (10 years, 4 months ago) |
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That doesn't look like E coli. Did you have a specific antibiotic to allow for only gram negative bacteria? RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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Amanita virosa
botanist by day


Registered: 12/04/11
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Re: Bench top myco filtration experiment. Complete! [Re: RogerRabbit]
#18891703 - 09/25/13 10:34 PM (10 years, 4 months ago) |
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Quote:
RogerRabbit said: That doesn't look like E coli. Did you have a specific antibiotic to allow for only gram negative bacteria? RR
I used a pure ecoli strain from the micro lab at the college where I work. I made the culture by pippetting one ml of sterile water into a agar plate of pure ecoli, agitating it then sucking it back out. I then added it to 2000ml of sterilized lysogeny broth (lb). Lb is ecolis favorite food. Incubated at 100 degrees for 24 hours and it clouds up with a gazillion ecoli. They go anaerobic quickly and have a very distinctive aweful smell. No antibiotics were involved. I can take some microscopy photos if I have time tomorrow.
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