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1. **Sterilization**     - Before sterilization (and closing autoclave door), ensure the pH of the first culture material bag entering the autoclave is >6.     - Adjust the timeline (water addition → material mixing → autoclave door closing) based on production factors (space, temperature, material temperature, pH).     - Confirm correct sterilization parameter settings before starting the autoclave.   2. **Cooling**     - No negative pressure or reverse airflow in the cooling clean area; positive pressure airflow direction: Cooling room → Autoclave rear door buffer room → Outside air.     - Positive pressure check methods: ① Light a lighter at the junction of areas with different purification levels (flame drifting from Area A to B means A is positive pressure); ② Use a differential pressure gauge (value >10 Pa).     - After 24 hours of cooling, bag core temperature should drop to 23°C (ideal for inoculation). Avoid post-inoculation condensation on bag walls in summer; keep temperature near upper limits in winter to promote germination.   3. **Inoculation**     - **Clean Area Entry**: Follow the manual for inoculation staff entering the clean area.     - **Key Precautions**:       ① Bag core temperature: 20–25°C pre-inoculation; inoculation room temp ≥23°C in winter (for rapid germination) & no condensation on bag walls in summer (to avoid temp difference issues).       ② Cleanroom standards: Inoculation room meets national Class 10,000 hygiene standards (positive pressure to outside); FFU meets Class 100 standards; inoculation buffer room & post-inoculation transfer room maintain positive pressure (entire cleanroom is positive pressure to outside).       ③ Liquid culture: Shake flask culture age ≤9 days, fermentation tank culture age ≤10 days; no foreign bacteria (confirmed by microscopic exam) pre-inoculation.       ④ Disinfection: Inoculation room disinfected with 75% alcohol or chlorine-containing disinfectants (used alternately).     - **Liquid Inoculation Process**:       Disinfect tools/hands with 75% alcohol; flame-sterilize inoculation gun tip. After inoculation, drain 1–2L liquid from front pipe.       Under FFU-purified air: Open bag lid → activate switch to transfer culture → close lid quickly.       Ensure full inoculation (avoid over/under/missed inoculation); prevent liquid spillage on bags (to avoid contamination). Inoculation volume: 35–50 ml per bag.   4. **Discussion**     - Deer antler mushroom mycelium grows slowly; even with liquid inoculation, it’s hard to quickly cover the medium surface. Thus, primary cultivation (colonization) must be in a highly clean room (designed/modified as clean room, managed per clean area rules).     - Besides space cleanliness, companies also focus on screening robust strains and developing supporting technologies.