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Inoculation of Hericium erinaceus

Inoculation is critical in bag production, requiring strict aseptic techniques. It’s best done early morning or night (to reduce contamination).

 1. Site Disinfection    - Inoculation can be done in a box, room, or plastic tent (must be sterile).     - Two-step disinfection:       - **First**: Clean the site 2–4 days in advance; fumigate with formaldehyde/sulfur 1 day prior, or spray with formaldehyde/phenol. Seal doors/windows for 12–24 hours.       - **Second**: After moving bags in, use aerosol kits (50g/kit; 4–6g per cubic meter). Light 30–60 minutes before inoculation to produce smoke.     - Note: Sterilization rooms with good sealing can double as inoculation rooms; poor sealing ones need a plastic tent inside.  

2. Strain Treatment    - Strain bags may have contaminants, so disinfection is required during inoculation (done with material bags).     - Steps:       - Operators wear medical gloves, disinfect hands with 75% alcohol.       - Wet strain bag cotton with alcohol; wipe the bag surface twice with 75% alcohol (or use 0.1% potassium permanganate: soak strain in solution, dry upside down).       - Use a blade to ring-cut 1cm deep, 1cm below the upper material surface; remove culture material and cotton plug.       - Cut the bag longitudinally, open it, and take out the strain.  

3. Inoculation Procedures & Methods   - **Operator/Equipment Disinfection**:       - Operators: Wash head/hands, change clothes; wear special workwear, hat, mask, and ultra-thin latex gloves.       - Equipment: Wipe with 75% alcohol, then flame-sterilize with an alcohol lamp.     - Inoculation Methods:       - **Tape-sealed ports**: Open one corner of the tape over the hole; insert a strain block, press slightly if protruding, and re-seal the tape. (1 bottle/750ml inoculates ~30 bags.)       - **Non-tape ports**: Wipe bag surface with 75% alcohol; use a smooth punch to make 3 holes (1.5cm diameter, 2–2.5cm deep), rotating clockwise when withdrawing to avoid air entry. Immediately insert a triangular cone-shaped strain block (matching hole size), press firmly to create a slight bulge (aids germination and seals the hole).     - **Post-inoculation**: Extinguish the alcohol lamp; move inoculated bags to a disinfected culture room; clean the inoculation site.