Auto-SummaryThe post outlines a new substrate method for growing coprophilous species, combining elements from various tek recipes. The basic substrate recipe consists of coir, BRF, vermiculite, and a tea solution. The casing recipe uses a mixture of peat and vermiculite. In-vitro jars are prepared with grey, leakproof lids and sterilized using steam. The jars are loaded with substrate, tamped down, and inoculated with liquid culture. The straw boss recipe is also provided, utilizing straw pellets for an all-in-one tray system. This method aims to minimize contamination by eliminating manure and grain spawn, and by using steam treatments to sterilize the substrate.
I have not grown using these recipes yet, I don't have the space available to do so at the moment; however, the concept is sound and should work as is but may need tweaking.
I had purchased the following spores to run this Manure/Coir/Verm/Brf substrate:
- Panaeolus Bisporus
- Pan Cambo var Thailand
- Pan Cyan var Estero
- Ps Subtropicalis var Xico (wild)
I'll leave this here for any interested members of the community to play with. I'm aware the following steam treatments were overkill but, I had to do my due diligence. The basic coir recipe should be sufficient but, I think the straw amendment recipe at the bottom of the post will be the better choice.
The substrate as it's currently conceptualized:
The following is a combination of features adapted from Baba Yaga's Manure Water Tek, Mateja's coir/BRF recipe, PF-Tek, V.L's EF-Tek, Violet's In-Vitro Bottle Tek, and Fahtsters Ziploc Tek.
Manure water will be used to hydrate the coir/verm/brf recipe to avoid the requirement for grain spawn and to take advantage of all ingredient's ability to be steamed at atmospheric pressure in the same way we would process PF-Tek jars in a pot.
The intention is to produce a new yet familiar method for fruiting intermediate to advanced coprophilous species using available materials and methods. Utilization of an "all in one" substrate will translate to less time producing sterile media by removing the necessity for spawn while simultaneously avoiding the inevitable disruption/weakening of the colony by breakage during the shaking/spawning phases.
Liquid culture will be the preferred inoculum for this method, with LI being second, and plate culture being last (in-vitro jars only). Inoculate with multispore solution at your own risk, which will be significant unless the cultivator has taken pains to produce axenic spore stock.
Basic Substrate recipe:
- 2 parts Coir
- 1 part BRF
- 1/2 part vermiculite
- 2.5 parts tea: by volume.
1 part = 1 cup, scale up as required. I used lots of water to produce the tea, adjust the ingredients to the amount of substrate being made.
Break up manure into the water, allow to "steep" for 1 hour to create manure "tea", strain with mesh screen, then use to hydrate the CVB substrate. Add the manure tea to the coir/verm first, and stir in the brown rice flour last.
Casing recipe:
- 50/50: peat/vermiculite
- Hydrated lime @ .5-1tsp/cup of peat
- Hydrate to field capacity with tap water.
In-Vitro Jars:
I prefer using the grey, Ball No-Leak lids with 1/4" hole and 29mm .3um synthetic filter disc combo but, polyfill stuffed lids should work just fine. Ball leakproof lids have a little more height than thinner metal lid rings or white plastic lids. The difference in thickness allows for a bit more room to dial gas exchange for both unmodified spawn jar applications and for FAE control of in vitro fruiting jars.
[1] Load substrate into wide mouth 750ml jars (without shoulders)(or glad rounds/4oz PF jars to top fruit);
[2] Lightly tamp substrate down to roughly 1.5-2" depth;
[3] Apply a thin 1/4" casing layer, tamp to level;
[4] Apply foil caps, steam for 60-90 minutes in a pot with a tight-fitting lid, or use your Presto without
the jiggle weight (you may sterilize at 15psi for 60 minutes if you prefer, but I wouldn't);
[5] Inoculate with your choice of liquid culture, liquid inoculant, or agar wedge once cooled;
[6] Colonize on a shelf;
[7] Back lids off to increase airflow to fruit;
Utilize 6500k light throughout. The resulting fruits will produce sterile fruits and spores for cloning and printing. Feel free to fill standard PF tek jars to top fruit in your choice of fruiting chamber.
These jars have been sitting since May 8th, with no sign of molds.
Bags:
Bags will be run using sufficient sub to be shaped into place in your preferred size of tray, much like what Faht does with Ziplocs.
[1] Before being fit into a tray the bagged steam pasteurized/sterilized sub will be inoculated with LC,
sealed, and then thoroughly mixed;
[2] Bags will then be formed into trays, leveled at 1.5-2" depth, then allowed to colonize;
[3] Once fully colonized the bags will be cut open to case, re-seal bags, and then allowed to colonize and
fruit in-vitro.
Straw Boss recipe:
 
I've tested these straw pellets in the past, they're not dense like other types of pelletized straw, sort of a looser compression.
This material hydrates to a similar consistency as expanded hardwood fuel pellets and can be hydrated in the same manner. Previous tests were to hydrate by Lipa tek, seal with an impulse sealer, then leave without inoculation on the shelf for 1 month to test for mold. SB pellets can be Lipa tek'd successfully so, pasteurization/steaming with MCVB will be no problem.
This is where the reader may ask: "Why not just use the manure".
Manure is "shitty" to work with, if it's not handled correctly it becomes an easy target for molds due to trace nitrogen combined with heavy bioload and complex structure. Coir, vermiculite, BRF, and straw pellets are far less susceptible to mold and bacterial blooms due to the manufacturing process that produces them. Using manure tea to hydrate the above materials produces a pseudo-manure substrate with beneficial microbes but without the added nitrogen in the biomass; The integral supplementation also removes the necessity for spawn or colony breakage.
Cereal grain spawn is subject to the same susceptibility to contaminations as manure because both substrates are nutritious organic materials with heavy bioloads and complex substrate structures.
By eliminating grain spawn, manure, and the requirement to weaken the sensitive colony through multiple rounds of breakage and recovery, both during the shake, and at spawning, the substrate will be overall less susceptible to contamination due to limited exposure to contaminant biolad, and breakage.
The manure tea introduces beneficial microbes while limiting places for pathogenic bacteria or molds to hide during steam exposure, making the substrate far less likely to contaminate.
Substrate recipe with added straw:
- 1 parts Coir
- 1 part Straw pellets
- 1 part BRF
- 1/2 part vermiculite
- 2.5 parts tea: by volume (to be adjusted).
1 part is equal to 1 cup. Scale up the recipe as required.
The above coprophilous species do not require excessive carbohydrate supplementation, so dial as necessary, the above quantities of brown rice flour should be more than sufficient; if anything you may need to dial it back rather than add more.
All-in-One Trays:
Fill with the above substrate to 1.5-2" depth, tamp/level, add peat casing 1/4"; steam, inoculate.
 
Gasketed PP5 container, flat lid for fixing hardware. I drilled 3 x 1/4" holes, 1 on each end for .2um filter stickers, one in the middle for a 3/16 nylon female Luer fitting:
  
A bulkhead luer adapter would probably be a better choice but I don't have any lying around. This will make the all-in-one (AIO) tray a closed system. I just need to get some male luer caps to close up the adapter:
Inoculating with LC from the center ensures that growth radiates outward from the middle. When I get around to trying this out I'll be drawing from a DIY bioreactor closed system with a check valve:
No spawn, no breakage, no shaking, no recovery. AIO pan tray can be steamed in a pot without sterilizing anything other than agar, LC, and syringe. Fruit in an appropriate chamber once finished colonizing.
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