Protocol

Tanahashi et al. – To obtain the basic artificial diets, agar powder (450 mg) (Nacalai Tesque, Kyoto, Japan), mycelial powder (100 mg), and deionised water containing the preservatives (15 ml) were placed in a 50-ml glass test-tube. Test tubes were autoclaved at 121 °C for 15 min.
Agar with wood only may serve as control.

To obtain mycelia of each fungal species, 5-mm diameter disks of fungal mat were removed from PDA plates, floated singly on 15 ml of aqueous potato dextrose broth (PD broth) (24 g/L) in 50-ml flasks (Sigma Aldrich, St. Louis, MO, USA), and incubated at 25 °C for 10 days. Residues (=mycelia) of fungal suspension by suction filtration were rinsed with deionized water several times and freeze-dried for 24 hours. Dried mycelia were ground with mortar and pestle to 0.5 mm mesh powder and then stored in a desiccator Tanahashi et al., 2009.
Sorbic acid can be used as anti-microbial agent.

• Sorbic acid (0.828 g)
• L(+)-ascorbic acid (1.0 g)
• 133 and sodium hydrocarbonate (1.1 g) in deionized water (1000 ml) (Tanahashi et al., 2009).

Antibacterial additives:
streptomycin sulphate and chlortetracycline hydrochloride, at three different contents of 30, 60, and 300 ppm.Tanahashi et al., 2009 Use no more than 5˚C when using antibiotics.
In Autoclave
Add ascorbic acid to prevent oxygenation of diets in the autoclave. Can add sodium hydrocarbonate to pre-autoclaved diets to raise the pH to ca 5.5.