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Mycofiltration: fungal filters

 
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I couldn't find too much about this online, but it seems really interesting and I just heard about it a few days ago so now I'm really curious. I just have a few kind of random questions, but it's not urgent or anything.

How would a mycofilter compare to something like a little reedbed for greywater filtration (like a tank filled with gravel or sand and aquatic plants)? Or am I creating a false dichotomy, here? Do they filter out different things? From what I've read mycofilters are more for things like bacteria and pathogens, while using plants would focus more on harmful chemicals than harmful organisms that could hurt you. Would the combined power of both of those purify water ridiculously well? I know some people don't like using greywater for edible plants. Do you think myco+plants would create water clean enough to eat the plants that grow in the filtered water.

This is just a simple mycology question, but how to the mushrooms survive in an environment where they are actually soaked in water, as opposed to just being in a humid area. I would imagine they wouldn't be able to handle being in a bag that's actually soaked with water, or literally inside a stream. Do the fruiting bodies just have to be above the water, and the mycellium just doesn't care or what?

Lastly, do you have to replace them every so often? A synthetic filter of some sort gets everything it's filtering stuck to it so you need to switch it, obviously. Do mycofilters act in the same way, or do they do something like "eat" the stuff that it filters out. I'd imagine that because it's a biological, living filter, that it would somehow interact with the things that get stuck to it as opposed to just kind of being a mesh of mycellium that catches junk. And if so, what does it do to them (probably another simple mycology question)? What is the byproduct of fungal respiration and consumption? Is it 02 like with plants?

Thanks in advance for any answers, permies!
 
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I don't know much about the mycofiltration questions in your second paragraph, but I do know that Paul Stamets has used king stropharia for mycofiltration and oyster mushrooms to clean up oil spills.
Aquatic fungi are rather rare in nature and almost none of them (as far as I know) form mushrooms. (But I may be wrong...) The mycelium of the "regular" (terrestrial) mushrooms I've studied cannot survive being submerged in water for more than a few days, as least as far as I can tell. They all need to breathe in order to survive.
The byproduct of fungal respiration is CO2. In other words, fungi breathe in oxygen and they breathe out carbon dioxide, just like animals.
I hope this helped answer your questions!
 
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Hello!

I have been passionately researching this topic recently.

First place to start is to encompass yourself in Permaculture design techniques, taught by Bill Molison and Geoff Lawton.
As mentioned, Paul Stamets is one of the leading mycologists in the world.
I suggest reading "Mycelium Running" and his other books "Growing Gourmet and Medicinal Mushrooms" along with "The Mushroom Cultivator" for any information on fungi.

I am currently constructing a 55gallon Mycofiltration system which I plan on taking to Burning Man in order to filter the grey water produced by my camp of 30 or so people.
I am also planning on giving a thirty minute talk and demonstration for other participants, on the possibilities of greywater reuse in arid environments, even when it's only for a 10 day camp site.

I have been taking photo's and will make it a point to share here.

What I happen to understand is:
Submerged mycelium will act as a cellular membrane filter, even if it considered "dead or dormant". The substrate used to grow the mycelium, along with the amount of mycelium grown in the substrate, will determine it's overall water capacity and flow.
Commercial greywater systems use Gravel/Sand/Carbon/Plastic and other sedimentiary systems along with Reverse Osmosis.
Living mycelium produces exudates which break hydro-carbon bonds, intended to break lignin in woodyplants. These myco-enzymatic-achohols and such are what have been shown to decompose oils and pesticides in a six month timeframe.

Comprehensive Assessment of Mycofiltration Biotechnology to Remove Pathogens from Urban Storm Water
http://cfpub.epa.gov/ncer_abstracts/index.cfm/fuseaction/display.abstractDetail/abstract/9645/report/0

Application is the main factor.

Can you use mycelium colonized substrates instead of plastic for a greywater system? YES!
Captain Planet always said "You have the power!"
 
Jay Shinn
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Danny Carm wrote:
Lastly, do you have to replace them every so often? A synthetic filter of some sort gets everything it's filtering stuck to it so you need to switch it, obviously. Do mycofilters act in the same way, or do they do something like "eat" the stuff that it filters out. I'd imagine that because it's a biological, living filter, that it would somehow interact with the things that get stuck to it as opposed to just kind of being a mesh of mycellium that catches junk. And if so, what does it do to them (probably another simple mycology question)? What is the byproduct of fungal respiration and consumption? Is it 02 like with plants?

Thanks in advance for any answers, permies!



Depending on the amount of sediment added to the filtration system it would need to be replaced in a relative manner of time. A Septic style baffle system will reduce sediment load on the filter itself. Most substrates are grown on organic materials which will decompose over time, the benefit I see here, is that it can go in your compost bin instead of the trash.

If your intention is to drip feed a garden, you will theoretically be creating a compost tea system which is fed by greywater from your house and or rain catchment system.

Mycelium breathes in O2 and out CO2, the same as us, their growth and fruit development is affected by CO2 levels.

I think I inadvertently answered some of the other questions in my first excited ramble
 
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Jay Shinn : Thanks for posting, could we have a few pictures posted here! This is a very important, serious subject and we need a few more people willing to post to let this
Topic/Thread grow ! Have Fun at Burning Man, There have been other types of grey water handling systems posted at Instructables.comin past years mostly
dehydrating types Again thanks ! Big Al
 
Jay Shinn
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allen lumley wrote: Jay Shinn : Thanks for posting, could we have a few pictures posted here! This is a very important, serious subject and we need a few more people willing to post to let this
Topic/Thread grow ! Have Fun at Burning Man, There have been other types of grey water handling systems posted at Instructables.comin past years mostly
dehydrating types Again thanks ! Big Al



First thing I did was start gathering materials for re-purposing that others were treating like trash.
Then I got at it like McGuyver.

As of this post... I have a 55 gallon drum on it's side with a spigot and a perforated pipe running the bottom, the same as a leechline or a inlet for a hydroelectric system in a ditch.
I plan on wrapping the pipe in screen and then laying in a layer of locally acquired red lava rock, and a whole lot of mycelium covered wheat straw. Sand or gravels could be used also, but I am trying to keep it rather lightweight for my travels.
I also thought about a layer of carbon(charcoal), but I would prefer to make it myself rather than purchase it.
On top I will cover it all inside the drum with a screened evaporation section for compostables to dry them out and segregate. Otherwise your filter is making blackwater while it makes filtered greywater.
Remember there are over 50 shades of greywater... lol

My goal is to at least by able to re-use campsite water for cleaning dishes and such, kind of like a stillsuit for camp, without the biowaste.

I started an album. I will keep uploading and try to take pictures as often as possible.

http://s109.photobucket.com/user/shinndragon/library/Mycofiltration%20GreyWater
 
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Hey Jay

I'm trying to get a visual of your system. So like are you running grey water from a sink drain (or whatever) into that black pvc with holes all down it and and letting it soak into straw which you have filled your blue drum with? Is thats whats up? couldn't tell from your 4 pictures, but that seemed plausible.
 
Jay Shinn
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Landon Sunrich wrote:Hey Jay

I'm trying to get a visual of your system. So like are you running grey water from a sink drain (or whatever) into that black pvc with holes all down it and and letting it soak into straw which you have filled your blue drum with? Is thats whats up? couldn't tell from your 4 pictures, but that seemed plausible.



Pictures are starting phases thus far, More to come when I actually have time from my crazy projects

Greywater or tap water will enter from the top opening via hose or 5 gallon bucket, I could do a pipe from the house but I rent, I do not own.
The water will go through layers of mycelium/straw, sand and gravel, before leeching into the pipe and out a spigot on the outside.
I intend to pre-filter out any large organic kitchen-waste material, but it could be used as an outdoor compost system also.

I am washing gravel, and growing out mycelium on pasteurized wheat straw.

I would like to simply be able to provide a way to re-use my camps dishwater and simple washings while camping in the desert, Dr. Bronners only or something similar obviously.
At first I was thinking "Dune" by Frank Herbert. Water is Life.

This could also be used as a worm-bin, to gravity feed worm-tea through the garden.

Possibilities are endless, but resources are limited.

Imagine your garden is your aquarium and your plants are your fish...
This system could act as a bio-filter for a pond or outdoor water-display.

On a larger scale it could be used to filter urban storm water runoff, and anything that leaves a farm or sewage treatment plant.
 
Landon Sunrich
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Alright I think I'm seeing it.

A portable grey water filter. I like where you're going. I might start playing with this one too. Please keep us all up to date on how it works out.

I'm pretty much in a position where I can work on crazy projects all the time -money being the primary constraint- that means I'm either in front of this #&*$ing screen looking for ideas or out in my yard doing something cheep and productive.

Really glad I have access to this forum as Ive been going it alone thus far and that can be very frustrating and isolating. Pretty sure regular cross pollination is necessary for the germination of ideas.

I've read Mycelium Running and I see the potential too. I have a lot of spawn going - well mostly waiting - you know, for the proper inspiration.

I like the idea of soil, fungi, and transpiring plants being the earths stillsuit.
 
Jay Shinn
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Landon Sunrich wrote:
Really glad I have access to this forum as Ive been going it alone thus far and that can be very frustrating and isolating. Pretty sure regular cross pollination is necessary for the germination of ideas.



I am also very glad to have this forum, and very glad you are appreciating it as much as I am! Planeteers Unite!
I also know the solo frustration of having an idea and not knowing how to explain it to someone else.
A friend recently told me "Do it for yourself, then it's art." - A.T.

If you have read Mycelium Running, I suggest watching every Bill Molison Permaculture design Course Video you can find. In order to find ways to APPLY design concepts.

I completely look forward to your ideas and inspiration, who knows, I may get inspired by them and then we will really be on a roll
 
Jay Shinn
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http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=%2Fnetahtml%2FPTO%2Fsearch-adv.htm&r=2&p=1&f=G&l=50&d=PTXT&S1=mycofiltration&OS=mycofiltration&RS=mycofiltration

Read the "English" parts only if you choose to stay sane, most if it will be in "Lawyers" terms.
 
Jay Shinn
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Right now I'm using my unit to de-gass evaporative chlorine from my tap water, before I irrigate my garden.
I see this as a bonus in many ways, because I am also rationing the amount of water my house uses on the garden.
I'm filling it up the night before with a hose and leaving an air-stone running in it, buried under the gravel.
I have not yet put in Mycelium/WheatStraw, but will be doing so soon for a first trial. I have until mid august, to have a complete unit.

So far, I'm simply applying Indoor Hydroponics concepts to my urban outdoor garden.
The drain spigot can easily lead to an evaporation system when i take it to Burning Man (Leave-No-Trace desert camping for 10 days).

I just uploaded a few more photo's of the air-line concept, which helps promote an aerobic environment.
http://s109.photobucket.com/user/shinndragon/library/Mycofiltration%20GreyWater
 
Jay Shinn
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Photo's !






All my grey water can now be mycofiltered! Removing soaps, fats and oils, before going to the evaporation system, or to be re-used for crude washing. ( for "Leave No Trace" camping)
Remaining water will come home to the garden and everything will be composted.

For a full-time installation, instead of a portable sink unit, one could simply pipe in water from the house.

I wanted to make a completely enclosed system so that nothing could escape during my adventures, this unit will be riding on a trailer.
Proud of myself for only using re-purposed materials
 
Landon Sunrich
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oh wow you really did go all mcguiver on that thing! I didn't realize you where looking to intigrate you're sink into your design. That's cool! Really cool! have you gotten a chance to test it yet? I have still be in the 'foggy intellectual brainstorming' phase but I still have more oyster spawn than I know what to do with!

also how are you getting your water to evenly distribute through the straw?
 
Jay Shinn
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Sink was a last minute idea and I went with it,
When you can get that feeling of "oh I could do this..." It's best to run with it

I also had that phase of 'foggy intellectual brainstorming' and found myself being a little held back by it, even though it gives a sane foundation for being crazy.

I didn't want to get it "dirty" before my adventure, but I did test the system concepts for my garden a while ago while in the 'still foggy trying to get out of fog' phase.

I opted to not worry about even water input distribution, but I totally thought all about that also. My half-assed conclusion, was "I've already made this thing cool enough".

Thoughts were that as the system gets used, areas that get "clogged" will simply cause water to move elsewhere.
I also presume that the filtering will be more "capillary" rather than forced. 5 gallons might be the most extreme amount of greywater to pass through it at any one time.
Also, the way I have it built, there will be a bit of "standing water" in the base before water is able to actually go out the spout.

In the end, prototypes are meant to be tested, but you have to build them first, to test them.

I know I just want to filter grey water for an evaporation system, but I like to go all out with mycofiltration as a means of demonstrating to others that there are possibilities.
 
Landon Sunrich
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Jay. I totally get that go with the flow thing.

I opted here for a really simple design. Two 5 gallon buckets and a drill. I nested the one with the holes drilled into the other. Obviously the top bucket (with the holes) contains the mushroom spawn.

I'm open to any and all suggestions for improving it! Mostly though I'm wondering if anyone can come up with some simple experiments so we can test these out! Have you gotten you're up and into action yet?

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Jay Shinn
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YES! PERFECT! AWESOME!

I have such a big smile right now.


Ways to improve are going to be found through trial and error depending on how you apply it.

I have been told filtration scales with surface area contact.
You could add a layer of gravel/sand or anything else like that to increase filtration if you felt the need to. Septics rely on sedimentation and baffles. Filtering fats and oils takes surface area and contact time.
Too much filtration will cause the system to rely on pressure from gravity. A slow filter which will be more efficient but less practical depending on the amount you pay pass through the system at any one time.
Possibilities are endless if you make a large settling tank prior to the filter acting as a resivoir/gravity feed.
I did a lot of research on septic tanks and how they function prior to researching greywater filtration.

Heres a commercial greywater filter for home use.
http://aqua2use.com/
Large scale units are pressurized tanks filled with sand.

My unit is packed up for Burning Man, I will be testing it intensively with a camp of 30 or so peoples greywater for 10 days.BM is a Leave No Trace event in the desert

The forum below kinda describes why I'm filtering even though I'm evaporating.
http://eplaya.burningman.com/viewtopic.php?p=334329
 
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Landon Sunrich wrote:
I'm open to any and all suggestions for improving it! Mostly though I'm wondering if anyone can come up with some simple experiments so we can test these out!



Oh, now you've gone and ruined a perfectly good bucket drilling holes in the bottom. You could have gone to your local garden center, got some 3-5 gallon size plastic pots from the recycle bin, and used those for the media holders. They usually fit perfectly in the top of a 5 gallon bucket.

Anyway, here's something to think about for a simple experiment. Use some dye or food coloring. Most dyes are organic molecules that will be metabolized by the fungi, and after they pass through the filter media, it should be decolorized. If you have one of the old type hand held light meters, you can put your before and after liquids in a glass and measure how much it was decolorized by the fungi. But be careful, this is getting awful close to real spectroscopy that they do in the chemistry laboratory, and if it's too much fun, you will want to sign up for an analytical chemistry course.

 
Landon Sunrich
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X-) *headsmash* A planter pot! why didn't I think of that!? The wasteful horror! sigh...

John - Food coloring is exactly what I was thinking. I just feared it sounded too childish and unscientific. Now that I know someone else thinks its worth a shot I'll get right on it. I just need to decide if I want red or blue mushrooms. No fancy gadgets here I'm sorry to say. But I'll keep my eyes out at yard-sales and the like. You best believe had I access to lab equipment and someone around who could teach me to use it I'd become a pain in their ass.
 
John Elliott
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For those of you who want to study up on this topic, I've compiled a reading list:

This paper talks about decolorization of azo dyes (like the kind used to dye blue jeans) by white-rot fungus, including the ever popular oyster mushroom.

In this study they went a step further and analyzed for phenolic residues from the dyes and found that the fungi were also metabolizing the possible dye residues.

But here's the jackpot, a whole 221 page book entitled "Biodegredation of Azo Dyes". That should keep you busy for a while.
 
Landon Sunrich
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I can see I'm going to need some brushing up on my 'science'. Chemistry was a first period class in my high school and man was I bored by the thermite at that hour. (seriously though science is almost always presented as dull and using highly specific technical language) I got through The pakistani study though. I'm actually rescanning it now as there were some things about 'glucose' and 'peroxidase' that I wanted to grasp a bit better and mill over.

I have changed my filter design slightly. I have added an inch or so layer of charcoal as center of a mycelial sandwich. I have flushed the bucket with fresh water several times. The natural color of the waste from the mushroom decomposition seems to be a reddish yellow or amber. I have decided to use blue food coloring which I will mix at a set rate per 2 liter of liquid. I will ad 2 liters of dye water at a time once every two hours for 10 hours resulting in 10 liters through the filter. I @ 12 hours I will then measure the amount of liquid that has run through the filter as well as make a subjective comparison of the coloration of that water with the untreated stuff. I have a feeling this will be the first of several experiments I run with this design. I will take some pictures of the process for peer review

Here's a quick look at the added charcoal layer. I put down another 3 inch layer of myciliated straw over it. It will be interesting to see what noticeable effects the biochar will have on the mycelium below it if any. Off to find some dye. Hopefully I'll have some results by tomorrow evening

I am also starting to get quite interesting in turkey tail. As it is another common mushroom around here which seems to favor alder
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John Elliott
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Just one caution about adding the charcoal. It is used in the lab to decolorize solutions of organic compounds that might contain bits of unwanted polymerized molecules. Big organic molecules (and this includes all of your dye molecules) find a nice resting spot on the high surface area charcoal, but the smaller molecule you are trying to isolate usually stays in solution.

I would suggest doing a control experiment with just the charcoal to make sure that it is not soaking up dye molecules. I take it the reason you added the charcoal is to provide surface area for fungal cells to colonize, which is great, but you don't want to introduce another way of having your input liquid getting decolorized that will confuse things.
 
Jay Shinn
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Comprehensive Assessment of Mycofiltration Biotechnology to Remove Pathogens from Urban Storm Water

http://cfpub.epa.gov/ncer_abstracts/index.cfm/fuseaction/display.abstractDetail/abstract/9645/report/0
 
Landon Sunrich
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So, My general thinking in adding the inch of biochar was to increase the effectiveness of the filter with as little change to the design as possible. The specific action I as hoping would be working is precisely what you supposed. I was hoping to use the micro-porous space of the char to soak up and hold more moisture basically as catching and holding medium for the exuded enzymes and fungal fingers to have access to. I am unsure how quickly liquid will pass through my filter and but I would imagine the longer they have contact with the fungus the more effective the filtration. It seemed like a biochar layre would provide for more surface area.

The EPA Final Report on Stamet's test stated that:

The second notable conclusion is that the permeability of mycofiltration media was generally in the range of 0.07 to 0.10 cm/sec —roughly equivalent to medium grain sand, indicating applicability for field-relevant hydraulic loading. Additionally, mycofilters demonstrated some capability to remove dissolved E. coli from flowing water, likely through an antibacterial mechanism. The final conclusion is that, as with other stormwater best management practices (BMPs), mycofiltration may be more effective against sediment-bound bacteria, and possibly achieve 100 percent E. coli removal.

But I am unsure if that ".07 to .1 cm/sec" is through saturated medium or unsaturated medium. Also wouldn't the size (diameter/length) of the filter mater as well as the volume of water? Also how exactly would one test for E. coli? that seems like it must be a common (cheep?) test.

Man now you guys have me wanting to dust of the old 'math' thing, draw cm marking on the side of my buckets. Measure some diameters and get serious with this thing.
 
John Elliott
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Landon Sunrich wrote:

The second notable conclusion is that the permeability of mycofiltration media was generally in the range of 0.07 to 0.10 cm/sec —roughly equivalent to medium grain sand, indicating applicability for field-relevant hydraulic loading. Additionally, mycofilters demonstrated some capability to remove dissolved E. coli from flowing water, likely through an antibacterial mechanism. The final conclusion is that, as with other stormwater best management practices (BMPs), mycofiltration may be more effective against sediment-bound bacteria, and possibly achieve 100 percent E. coli removal.

But I am unsure if that ".07 to .1 cm/sec" is through saturated medium or unsaturated medium. Also wouldn't the size (diameter/length) of the filter mater as well as the volume of water? Also how exactly would one test for E. coli? that seems like it must be a common (cheep?) test.



Here is a student lab that I lifted off the 'net that takes you through the basics of testing for bacteria. Realize though that there is a difference between testing for the amount of total bacteria by a centrifuge or turbidimetric method and doing something more specific for a particular species or strain, such as an immunoassay.

The centrifuge method is probably quicker for you to use since a turbidimeter takes time to build and calibrate, while for a centrifuge, all you need is a rope and a paint bucket. Get yourself a block of styrofoam or other material that will sit in the paint can and prevent your centrifuge tube from moving. Small bottles with screw caps that are taller than they are wide make good centrifuge tubes. Fill it completely to the top with your sample you want to assay for bacteria and carefully put the cap on, trying to minimize any air left under the cap. Now put the bottle in the styrofoam block in the paint can and secure the lid with a few hammer taps. You don't want the lid flying off in the next step. Now tie a secure double knot on the bail of the paint can and you are ready to centrifuge.

This next step is going to give you a workout. Start swinging the paint can in a circle and work up to a comfortable speed, not as fast as you possibly can, because you need to keep this up for a while. It's not the peak amount of "G"s you can get the paint can to pull at one time, but "G"s times time. Ten minutes should be enough time to get a decent centrifugation. Now go to it. Set the timer and keep it up, swing that can at a constant speed.

After your centrifugation, carefully remove the centrifuge tube from the paint can and see if the supernatant (the liquid at the top) is clear. If it is the least bit turbid, you didn't swing long enough or fast enough. If it is clear and you can see crud on the bottom of the tube, that is what you are after. Carefully pour off the supernatant, trying to disturb the crud as little as possible and then you are ready to move on to the weighing step of the lab.

So much for the gravimetric method of bacterial assay. If you want to MacGyver together a turbidimeter, we can do that too, but it involves more of a treasure hunt beforehand.

You are right to question the flow rates that they mention. There are indeed many other variables. So many in fact, that just distance divided by time doesn't tell you very much. It is a heterogeneous chemical reaction and what you really need is some sort of cobbled together measure that tells you the amount of contact time between the liquid phase and the solid phase upon which the catalyst (the fungi) is held. Generally, the smaller the pore size in the solid media, or the higher the surface area (this is why the biochar helps), the longer contact times that you end up with.
 
Landon Sunrich
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John,

Am I following you correctly in that I should be able to make a gross comparison of total bacteria in several wet mass samples using visual approximation all while spinning like a Sufi?

I'll try it.

What am I looking for? "cloudy?" How still are we talking about needing this wet mass needing to be? Like could I just stuff some socks and shirts in with it or do I need to carefully trace a Styrofoam* cutout?

*the red line of knowing suggested I capitalize that last one (TM)
 
John Elliott
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Yes, the technical term for "cloudy" is "turbid" and the turbidimeter measures exactly how cloudy a liquid is from suspended particles or bacteria. Here's what the EPA has to say about turbidity and why it is important. Unfortunately, turbidity is only half of a measurement, as once you have a number for it, the next question is "great, now what's the size of these particles that are making the water turbid, and are they clay or bacteria or something else?" As long as you are pretty clear on what is making the water turbid, like say E. coli in water passing through a mycelial filter, then you are justified in making sample-to-sample comparisons.

Sure you can use cloth wadding to immobilize the tube inside your makeshift centrifuge, but a solid piece of styrofoam/cardboard/wood makes it even less likely to shift around in the can while it is spinning.
 
Jay Shinn
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I'm back from the desert!

11 days of Leave-NoTrace Camping with a camp of around 30 people.

All the camps greywater was passed through my Mycofiltration system prior to evaporation. Equivalent to about 100 gallons of greywater from dishes, footbaths, and general cleaning.
The filter also got to deal with pounds of playa dust.

It never clogged and no particulate came through. Mission success.

Paul Stamets gave me a hug.
 
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very cool!
 
Jay Shinn
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Read Me!

http://fungi.com/pdf/articles/Fungi_Perfecti_Phase_I_Report.pdf
 
Danny Carm
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Jay Shinn wrote:Read Me!

http://fungi.com/pdf/articles/Fungi_Perfecti_Phase_I_Report.pdf



I actually saw that a few days ago! It's awesome that they were able to do such an in depth experiment.

I kinda forgot about this thread but I've been getting back into mycology and finally got around to reading Mycelium Running I just had a few more questions. For your system you took to burning man, what strain of mushroom were you using? And how do you keep a fully colonized bunch of straw from fruiting? Especially if you're regularly running water through it? I know they like sunlight to help them know it's time to fruit, but from what I've heard, it's not 100% necessary. Did you end up with mushrooms in your filter?


ALSO, for everyone in this thread, there's an awesome youtube channel/website called Radical Mycology (http://www.youtube.com/user/radmycology) with a lot of excellent content for growing mushrooms for mycoremediation. I just ran across it the other day, hopefully you guys might find it useful and interesting.
 
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