B.3.1. TIMELINE

B.3.1.
Transparent pot with a draining system

Run through: 9. 08 –
Total batch weight: 2.4kg;
Bokashi fermentation proportions = 1:3 → 600g Bokashi + 1.8kg HOW;


HOW chopped into small pieces with a knife; aerobic fermentation.

On 21.08.2019 half of this batch was put to dry and incubate at 72 degrees Celsius. The other half was left to ferment longer in aerobic conditions. After developing mould, this second half was thrown away.

Experiment 28. 08. 2019: a repetition experiment


⌕ Will repeating the same experiment lead the same results?

⌕ Does HOW material structure change when HOW and the mycelium spawn are homogeneously mixed or layered?

In the weeks leading up to this experiment, HOW samples have not been developing very well. This experiment focuses on verifying if repeating the conditions of successful samples will give the same results. Additionally, mycelium forming direction and structure are tested.

Day 01 into the experiment. All samples. Left column: homogeneously mixed. Right column: layered samples. In all the pictures, the samples are arranged in the same way.
Day 05 into the experiment.
Day 13 into the experiment.
Day 15 into the experiment. Samples ready to be dried. Samples B.3.2.1A (mixed), B.3.1.1B & B.3.2.1B (layered) failed.
Day 15 into the experiment. Example of one of the samples.
8 medium Petri dishes were prepared, using HOW from all dried batches laying around at this point, in pairs. All 'A' samples were mixed, meaning that the ganoderma mycelium spawn is mixed homogeneously with the dried HOW. All 'B' samples were layered, which means that the mycelium spawn is laid at the bottom of the dish separate from the HOW substrate:



The batch samples from B.0.1 and B.1.1 grew the same as before. B.3.1 and B.3.2 were newly introduced and behaved differently in the repetition conditions. This could have to do with the composition of the dried HOW.

Mixed samples became slightly curved towards the top, while the layered samples formed flat. A cause of this could have been that mycelium always forms its net upwards, thereby constructing a denser mass and tension in the upper part of the dish.

Experiment 16. 09. 2019: colouring attempt 03


⌕ Can the HOW material be coloured?

⌕ Which pigments can be used without being decomposed by the fungi enzymes during mycelium forming?

This experiment focuses on testing if different colour dyes preserve during the mycelium forming stage, and if colour dye hinders mycelium forming when the HOW substrate is present.

15 medium Petri dishes were prepared; 10 dishes containing HOW substrates from all batches laying around at the time, and 5 control dishes. The red and blue silk dyes were poured on top of the dishes after the mycelium spawn and the HOW were mixed. As yellow dye, turmeric powder was mixed homogeneously in the mycelium control dishes.



Day 01. Red control & all samples
Day 01. Blue & yellow controls
Day 04. Blue & yellow controls
Day 09. Red control & all samples
Day 16 of the experiment. Examples (left to right): samples B.3.1.2B, B.3.2.2A, B.3.3.1B & Yellow B
The red silk dye preserved during mycelium forming in most situations.
The blue silk dye was decomposed during mycelium forming, the colour dissapeared until the end of the process.
Of the 3 yellow dye control dishes, the C sample didn't grow.

This experiment concluded that colouring the HOW material is possible but the exact conditions in which this must happen are yet to be researched.

Experiment 9. 10. 2019: 3rd pH experiment


⌕ Does raising the pH of fermented HOW influence the course of mycelium forming?

Previous to this experiment we tried to see how raising the pH of the fermented (and acidic) HOW substrate will influence the mycelium forming phase. All previous pH experiments were inconclusive: we couldn't assess the extent to which (1) the substances used to raise the pH (baking soda and dishwashing detergent) or (2) the default substance composition of the HOW substrate inhibit mycelium forming. Dried fermented HOW substrate has a pH of approximately 4.

In this experiment two new substances are used to raise this pH gradually: disodium phosphate (Na2HPO4) and sodium hydroxide (NaOH), both capable to significantly raise pH levels. 4 solutions were prepared using these substances:
Solution pH=8.44 → Na2HPO4 + water
Solution pH=9.43 → Na2HPO4 + NaOH + water
Solution pH=10.89 → Na2HPO4 + NaOH + water
Solution pH=11.81 → Na2HPO4 + NaOH + water

To assess if the default substances in the composition of dried fermented HOW also play a role in mycelium forming, we chose 2 batches to experiment with: B.0.1. (best performing batch), and B.3.1. (one of the worst performing batches).

26 medium Petri dishes were prepared. Dried HOW was soaked in water or the solutions indicated above. We expected that the pH would keep changing over time, however these initial measurements are used to indicate the gradual pH difference between the samples. Samples 'C' and 'D' are layered, which means a layer of ganoderma mycelium spawn is laid at the bottom of the dish and another layer of HOW substrate (with varying pH values) is laid on top:



Day 01. All samples
Day 10 of the experiment. Serie B.0.1. Top: front of the samples; bottom: back of the samples. Positioning (left to right): [top line] B.0.1.9A, B.0.1.10A, B.0.1.11A; [bottom line] B.0.1.9B, B.0.1.10B, B.0.1.11B
Day 10 of the experiment. Serie B.3.1.3. Top: front of the samples; bottom: back of the samples.Positioning (left to right & top to bottom): B.3.1.3A, B.3.1.3B, B.3.1.3C & B.3.1.3D
Day 10 of the experiment. Serie B.3.1.4. Top: front of the samples; bottom: back of the samples.Positioning (left to right & top to bottom): B.3.1.4A, B.3.1.4B, B.3.1.4C & B.3.1.4D
Day 10 of the experiment. Serie B.3.1.5. Top: front of the samples; bottom: back of the samples.Positioning (left to right & top to bottom): B.3.1.5A, B.3.1.5B, B.3.1.5C & B.3.1.5D
Day 10 of the experiment. Serie B.3.1.6. Top: front of the samples; bottom: back of the samples.Positioning (left to right & top to bottom): B.3.1.6A, B.3.1.6B, B.3.1.6C & B.3.1.6D
Day 10 of the experiment. Serie B.3.1.7. Top: front of the samples; bottom: back of the samples.Positioning (left to right & top to bottom): B.3.1.7A, B.3.1.7B, B.3.1.7C & B.3.1.7D
This experiment tried to assess mycelium forming in correlation with the pH value of HOW, the substances in the composition of HOW, and the type of growing (mixed or layered). All samples were left to grow at 25 – 30 degrees Celsius, followed by being dried and incubated at 72 degrees Celsius. All samples in this experiment were successful.
Generally, samples with higher pH values tended to grow better and slightly faster. Layered samples grew slower than homogeneously mixed samples. Notably, B.0.1. didn't perform better than B.3.1., which may indicate that the HOW composition doesn't matter in the mycelium forming stage.
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