MYCELIAL EXPLORATIONS

By Paige Perillat-Piratoine
Steadily now, research is advancing its knowledge about the central role of fungi in our biosphere. This comes with an incredible surge in mycological projects worldwide. From in depth studies led by universities to small-scale citizen science and industrial farms, the potential of this understanding is being explored; and the applications are as endless as human creativity.
Projects include breaking down toxic waste, crafting a wide range of biomaterials and exploring the medicinal applications of this kingdom. Essentially, these projects contribute to the start of narrative towards organic architecture, remediation and medicine, and many other things that may not have names yet. Here emerge imaginations of a future reality that can go both ways: either depicting symbiotic human materialities or seriously dystopian scenarios.
One of the projects is happening here, at Greenlab:
Turning organic waste into a London based micro-medicinal mushroom farm
i.e.
Growing Cordyceps Militaris on Spent Brewery Grain.
So why an urban Cordyceps micro(micro!) farm?
Possibly because of the story it tells. This is an incredible fungus: an entomopathogenic fungus. In other words, an insect parasite. In Nature, when its spores manage to penetrate a suitable host insect, they weave mycelium in and around its organs, eventually reaching the brain and ultimately hacking its body. It then can direct the insect to move to the most suitable place for the fruit body (the cordyceps mushroom) to sprout out of its host. See David Attenborough narrating the process: https://www.youtube.com/watch?v=XuKjBIBBAL8
Despite this cruel process, it turns out the cordyceps mushroom has various chemical components that benefit human health. There are many types of Cordyceps, but the one I am growing here at Greenlab is Cordyceps Militaris. Somewhere in 1980, someone managed to grow this variety on something else than insects, opening the possibility for much a more industrial processing of this fungus. Now, especially in Thailand, Cordyceps farms are the norm. But not in London; Not in Europe in fact.
Yet cordyceps is an extremely valuable mushroom. It has been proven (after being used ethnobotanically in China for at least a thousand years) to help in a number of ways including increasing endurance and stamina, boosting the immune system, improving respiratory function, improving heart function, etc.

And it can grow on a abundant urban waste product: spent brewery grain. When grown in this way, it seems the cordycepin content (the primary active phytochemical in cordyceps) is higher than on other grain. (see https://www.researchgate.net/publication/274381565_Cordycepin_production_by_Cmilitaris_cultivation_on_spent_brewery_grains ).
This means there is the potential to create an interesting closed loop industry – By growing cordyceps militaris on brewery bi-product we are creating an urban biocycle*:
Accordingly, I intend to demonstrate the feasibility of this urban biocycle (the micro-medicinal mushroom farm) as an instrument of contemplation highlighting the potential of the fungal world to engage with current issues at a much bigger scale, mycelium being nature’s silent and still largely undiscovered decomposer.
This project is also intentionally London Based: it intends to tackle the place specific challenges of farming in the unique area of London;: it will measure the conditions provided by the city of London: what obstacles and potentials exist here? This may help the general region of climate-similar Europe to understand what challenges will present themselves in semi-constant rainy, grey conditions, and perhaps come up with solutions.
*Biocycle – the process whereby a waste material is diverted from the trash and transformed to create a new product which can be reinjected into the economy. “Biocycle” refers as much to the process as to the finished product.
****Disclaimer: I have no background of academically studying mycology/biology or even science. This project is the result of extensive personal research, self-teaching and experimenting – As such, the reader may know more or less than I do – this is of no importance. The style of the article simply intends to be accessible whilst open-sourcing new or existing ideas.
NOW THE PROCESS.
Preliminary Information:
This cordyceps grow is based on methods described by the company Mycosymbiotics in the following resource: https://www.etsy.com/uk/listing/504870723/cordyceps-cultivation-ebook
It involves a relatively beginner friendly process of procuring a liquid culture, inoculating a substrate, incubating the culture and fruiting the culture. This stands in contrast with more complex mushroom cultivation which can involve several more steps of feeding a culture various nutrients before fruiting it – resulting in higher yields.
This entry will be supplemented as time progresses, and results are achieved; keeping in mind that a mushroom growing timescale has its own slow pace, hence results will be released at the same slow pace.
This guide is intended to be accessible and relatively easy to follow – It will eventually boil down to a single method with very specific ( and place-specific) indications.
Having now started experimenting on my own, here is how I understand the process.
****Disclaimer: This research is still ongoing, as a result this document can only be considered as experimental field notes, and not a finished guide of the cultivation process.
1ST STEPS: SETTING UP A SUITABLE SPACE –
Space needs are divided in 3 – and roughly correspond to the different phases of cultivation:
—> The Sterilization and Inoculation space: This is where one pasteurizes and inoculates the substrate – This space will ideally be a closed environment where one can restrict airflow (this limits the possibility of contamination), and will have very easily cleanable surfaces. A kitchen is usually suitable; This is a temporary space: i.e it you will only use it for a few hours and then it will be clear again. Unless you want to store your equipment there.
—> The Incubation space: This is where you will put your inoculated jars to be colonized. It needs to be pitch dark and between 15-23 °C
—> The Fruiting space: Once your jars are fully myceliated, you will transfer them to light in the fruiting space. Here too temperatures need to be constant between 18-21°C. In this phase, if you are successful, you may see fruit bodies (mushrooms) emerging. From tiny pinheads to full grown shapes.
Once you have figured how much space you have and where you will execute the different phases, you can start
PROCURING YOUR MATERIALS.
Your base grain should be organic – Mushrooms are potent bioremediators and have a tendency to take up what is in their soil, if your grains are full of pesticides, it is likely that your cordyceps will be as well
Here is what I have used so far:
Substrate :
-Spent Brewery Grain
-Sucrose
-Potato starch
-Azomite
-Yeast Extract
– |
Materials :
-Cordyceps liquid culture syringe
-Jars with lids
-Pressure cooker
-Food dehydrator
– Polyfill/cotton
– High temp silicone sealant
-Labels + Pen
-( Clear box & black box) |
Sterile requirements :
-Surgical gloves
-Alcohol spray bottle
-Lighter
-Surgical mask/or clean scarf |
One-off tools needed: Drill (for making holes in the jars)
SOURCES USED IN THIS PROJECT
-> Organic Brown rice:
https://www.amazon.co.uk/gp/product/B007CX0BCK/ref=oh_aui_detailpage_o07_s00?ie=UTF8&psc=1
-> Sucrose (Sugar) : As Nature Intended
-> Yeast Extract : https://www.amazon.co.uk/gp/product/B0752WKWRF/ref=oh_aui_detailpage_o01_s00?ie=UTF8&psc=1
– >Potato Starch : Holland & Barrett
Azomite : https://www.ebay.co.uk/itm/AZOMITE-pure-TERRALBA-100g-germination-roots-flowers-fertilizer-natural/253324784559?ssPageName=STRK%3AMEBIDX%3AIT&_trksid=p2057872.m2749.l2649
Jars: https://www.jarsandbottles.co.uk/glass-bonta-jar-314ml-70mm-twist-off-cap
Pressure cooker: http://www.argos.co.uk/product/5550067
Silicone: https://www.amazon.co.uk/gp/product/B071JDRT8P/ref=oh_aui_detailpage_o00_s00?ie=UTF8&psc=1
Clear Box: https://www.amazon.co.uk/gp/product/B01N3RAFYD/ref=oh_aui_detailpage_o04_s00?ie=UTF8&psc=1
Black box: https://www.amazon.co.uk/gp/product/B001O3IE96/ref=oh_aui_detailpage_o05_s00?ie=UTF8&psc=1
Lighter : Tesco
Alcohol Spray: https://www.amazon.co.uk/gp/product/B074PTMZ8X/ref=oh_aui_detailpage_o08_s00?ie=UTF8&psc=1
Gloves: Pharmacy
Labels: Ryman
Cordyceps Liquid Culture from: Ebay : UK Mushrooms supplies.
Polyfill: https://www.amazon.co.uk/Dempsey-Designs-Factory-Minicraft-Filling/dp/B000WNZ1SQ/ref=sr_1_1?ie=UTF8&qid=1524759546&sr=8-1&keywords=polyfill
**** I’ve chosen to use an electric pressure cooker – While it does not get to the 15psi pressure recommended by cultivation practices, I have found it worked for my purposes. Using it is easier as it means I fill it with jars and water, press a few buttons and can relax/do something else for 2 hours while things sterilize/pasteurize.
**** The size of your pressure cooker and size of jars matters : the jars must fit in the pressure cooker (and not be too squeezed together) – do not invest in one or the other unless you know they are compatible.
**** The liquid culture from UK mushroom supplies has not fruited yet and seems to be having trouble. It may not be a good one. I’ve now switched to cultures ordered from the USA @ https://www.etsy.com/uk/shop/Mushroomrevival
MAKING YOUR EQUIPMENT.
TO MAKE YOUR JARS:
Order your jars and take the lids off. Drill two holes in each lids of approx 5 mm wide: one more central one to the side. On the central one you’ll apply the silicone (squeeze out the silicone on to the hole, cover the hole on both sides, and leave to sit for 24hours to dry and settle) : this will be your injection port for the syringe containing the liquid culture.
In the other you’ll fit through some of the polyfill which will serve as an air filter. Cut of anything that sticks out too much.
See following video for visual instructions. https://www.youtube.com/watch?v=PC-o_pWIU1k
TO MAKE A GLOVEBOX:
Gather a clear box, and 2 pieces of approx 10cm diameter PVC pipe (3/4cm long). Gather a sharp hunting knife and a lighter. Trace your holes on the glovebox (use your pvc pipes and trace their outer rims with a sharpie) where you’ll be putting in your arms. Heat your knife till it is red, and start using it to melt the plastic following your traced lines. Slowly melt along the lines till you have created the arm holes, and punch the center piece of plastic out (reheat the knife as many times as needed, as it needs to be scorching so the plastic does not crack). Fit the pipes into the holes and apply sealant around the edges. Smooth the sealant as much as possible and let dry for a day. I have not put gloves on my box as I am praying it with disinfectant before I inoculate and wait for it too settle- I have not gotten contamination yet.. If you want to be more thorough Watch this video: https://www.youtube.com/watch?v=lSwJosoNbZY
***** It’s not clear if a glovebox is really necessary. I’ve done my second batch without and I have no contamination.
PASTEURISATION/STERILISATION.
Once you have chosen your spaces, prepared your jars and have gathered all you materials and equipment, You can inoculate your first batch.
I’ve started experimenting with brown rice, to start simple and get the hang of things;
PASTEURIZE
→ Go to your inoculation space and close all the doors and windows and clean it thoroughly with disinfectant products.
→ Grab your Jars (I do 6 jars in one batch, as only 6 fit in my pressure cooker)
→ Take ¾ cup of brown rice, and divide equal amounts into each jar
→ In a separate (pouring type) container add 1.5 cups of water then add 1 tsp sugar, ½ teaspoon of potato starch, ½ teaspoon of Yeast extract and a pinch of Azomite: this is your nutrient broth.
→ Divide that broth between your 6 jars
→ Close your lids
→ Put in your electric pressure cooker on Stew setting for an hour.
→ release pressure on your pressure cooker by turning the valve (use a spatula) and wait for fumes to be completely expelled
→ Take your jars out, put them on your counter to cool down (with sufficient space between them so they cool faster)
→ In about 2 hours they should be cool.



INNOCULATION
→ During the pasteurization time you will have taken a shower, put on clean clothes as well as put on a mask/scarf in front of your mouth and nose (your breath is Full of competing bacteria)
→ Put on your gloves
→ Wipe gloves with alcohol.
→ Take your liquid culture syringe from your fridge 30 min before planned inoculation,
→ Build it up ie. put the needle in & Shake the mycelium to give it oxygen
→ clean the needle with alcohol
→ use the lighter to scorch the needle red & let the needle cool down (repeat before each jar)
→ punch the needle into your silicone injection port and press out approx 1-2 ml (depending on the number of jars you have)
into a jar in a circular motion (this ensure you distribute the mycelium somewhat across the substrate)
→ remove syringe
→ sterilize needle again and inoculate next jar.
→ repeat until all jars are inoculated
→ Label your Jars (Name of strain and date Inoculated)
→ Put them into their incubation space.



****Note on required State of mind: as a sterile procedure novice there is a need to clear the mind before going about this process. Why? Because there are so many little steps and because of what you are trying to avoid. In the air around you there are millions of spores, bacteria and general potential contaminants. There is a perception, when you are not in a full lab condition , and whether it is true or not, that your movements need to be slow and steady, there needs to be a meditative state of mind really, because any abrupt movement causes to much air movement which in turn affects your sterility levels and so may contaminate your culture- wasting lots of your time in the process (that said contamination can be beautiful)
Bring your attention to the fact there are several tiny things to remember, they might be simple but their accumulation is mind-consuming at the beginning. Note to self : Keep trying it will eventually engrain into body processes.
INCUBATING (Colonisation period)
As soon as you are done inoculating, put your jars in your incubation space. Check in one or two weeks; You should expect your substrate to be white, covered in mycelium and fluffy looking. You can expose it to light as soon as the surface is mostly white and fluffy.
Remember not to disturb this space too much, to keep it dark and at a stable temperature: between 15 and 23°C.
To keep track of my temperature and light levels during the colonization and fruiting periods, I used a smart citizen kit which you can plug in and place next to your cultures to understand precisely the fluctuations in temperature and light your culture will be experiencing. For my first two batches, this was very useful as it helped me understand it had gotten too cold or too warm on some days, indicating I might have damaged my culture. You can find the smart citizen kit here https://smartcitizen.me/


FRUITING.
So once the top of the substrate is fluffy and white – Put it in your fruiting space. It should be in indirect sunlight or under a lamp for about 12 hours a day (I prefer a white type LED lamp on timer as it is more reliable in London where I feel there is not enough light for the culture to be happy- Online cultivators recommend red and blue LED light strips for higher cordycepin production)
Keep a stable temperature between 18 and 21°C. (This is tough. I might have to build an incubator for better results)

TROUBLESHOOTING.
The Mycelium looks scraggly:
It looks like this:



But a healthy cordyceps mycelium should look like this:

The temperature may not have been constant enough or there might not have been enough light. One comment from Cordyceps Cultivation group “high temperature make the filament damaged so you should put on temperature 23, for recovery of healthy mushroom and then shock 18-23 temperature in 7 days. After bringing the temperature to 23, increase the light to 700-1000 lux 12h per day, the moisture increased by 80-90%. Cordyceps mushrooms will sprout. good luck.”
Bad Culture Theory: Antony Gandia “I didn’t find any good EU supplier as well, I got all my strains from India and Vietnam, which most probably come from Thailand suppliers. The Vietnamese strain I got from two awesome guys operating in Quang Ngai rocks like a hurricane (see picture). It’s very important to get a viable strain that comes from a recent spore mating, there’s a lot of in vitro work to do to keep these species going on. If you have the money, the time and the chance, I strongly recommend to travel to South-East Asia to get good strains and to learn the methods directly from operative farms, otherwise you will most likely receive old and senescent cultures from people that just stock them”
LIQUID CULTURE.
When you make a liquid culture you take spores, transfer them to a liquid nutrient solution and let them colonise that liquid solution. This is what you can then inject into a non-liquid substrate for mushroom growth.
For Cordyceps: Just prepare a light ‘beer’ broth using cereals or potato, filter it, add brown sugar, and sterilize in a jar. Inoculate with a proper culture or spores and shake everyday to give in a bit of oxygen.
HARVESTING: AWAITING RESULTS
DRYING: AWAITING RESULTS
PRESERVING: AWAITING RESULTS
STILL IN PROCESS: FINAL SUGGESTED RECIPE ( SPENT BREWERY GRAIN), FINAL MATERIALS USED AND SOURCES- FINAL TIMESCALES AND ESTIMATE ON 6 JARS
RESSOURCES
Pdf explaining mycology language:
MYCOLOGY LANGUAGE ( by Paige PP)
Cultivation Tracking Forms (by Paige PP):
https://docs.google.com/document/d/1p67s48-i8szd_svd0ts9TIP0vhaL8rY6SZP_crhs2V0/edit?usp=sharing
Facebook Cordyceps Cultivation Group:
https://www.facebook.com/groups/1449554748397163/
Research on spent brewery grain:
https://www.researchgate.net/publication/274381565_Cordycepin_production_by_Cmilitaris_cultivation_on_spent_brewery_grains ).
Cordyceps cultivation E-book:
https://www.etsy.com/uk/listing/504870723/cordyceps-cultivation-ebook?ref=shop_home_active_2
FORUM DISCUSSION
“Do you incubate in complete darkness until 100% and what’t your process for fruiting? 12/12 light cycle with a drop in temps?
Terrestrial Fungi I waited until 100% (colonisation) for light exposure this time, but it slowed me down in the long run, this next round I’m exposing to light as son as the top surface is mostly covered.
Sukoom UT We, 12-14 hrs @ ~1200lux fluorescent or day light, after pinning 800 lux with blue or white were enough”
LEFT TO DO
Harvest & use instructions
Left to do after successful trial ? Look into commercial techniques to maximise yields, medicinal contents, and ergonomize processes.
Travel to Thailand, learn from local farmers to understand equipment and materials needed?
Opportunities ? A larger farm in London? Why not ? Need for a mycologist/ microbiologist to truly develop commercial strains.