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Inoculation must be performed under sterile conditions in the inoculation room or inoculation hood. The specific procedures are as follows:

1. Prepare High-Quality Cultivated Spawn

Bottled spawn is preferred. The mycelium should be dense, robust, and uniform, free of foreign bacteria and pests, and without fruiting bodies.

2. Disinfection

Scrub the outer surface of the cultivated spawn with a fungicide such as 0.25% Sanisol solution, 0.2% potassium permanganate solution, or 0.1% carbendazim. Burn the bottle mouth over an alcohol burner. Sterilize the inoculation equipment as well. Fumigate the inoculation box or room with an aerosol disinfectant or spray with a fungicide to achieve a sterile inoculation environment. This ensures that no foreign bacteria are present during inoculation.

3. Inoculation

Inoculation should be performed when the temperature inside the bag drops below 25°C. Vaccinators should change clothes, disinfect, and air-shower before entering the inoculation room. Ideally, two people should work together: one to remove the seed and inoculate it, while the other opens and seals the bag. The specific procedure is to heat the inoculation tool over the flame of an alcohol burner. After cooling, scoop out the old culture on the surface of the bottle mouth. Remove the lower layer of culture and transfer it to the bag. Ideally, the added culture should completely and evenly cover the surface of the material. Then seal the bag.

Typically, 750g of culture can be used to inoculate 20-25 bags, and 600-800 bags can be inoculated per hour. After inoculation, the transfer baskets can be directly transported to the culture room using a dedicated cart. This way, from bag sterilization to shelf culture, several intermediate processes are carried out within the baskets, reducing workload, production costs, and minimizing contamination of the culture medium in the bags.