The cultivation of Agaricus blazei three-grade strains typically relies on wheat kernels as the primary raw material. However, in actual production, high pollution rates or abnormal growth patterns are frequently observed. These issues often arise from non-standardized strain production practices. To address this, detailed specifications and operational techniques have been developed to guide farmers and agricultural workers.
1. **Raw Material Preparation**: Cow dung is first sifted and mixed with water in a 1:1.2 ratio. The mixture is then fermented and turned daily for about 20 days. Once the cow dung turns brown and emits a normal fermented aroma, it is dried and stored for later use. Wheat grains are thoroughly soaked, drained, and placed in a shaded, well-ventilated area. When the outer layer is dry, they are lightly misted to maintain moisture. After 24 hours, sprouting begins, and when the sprouts reach 1–2 cm in length, the grains are ready for use.
If the wheat has been worm-eaten, it can still be used, but the holes left by insects may cause starch to leak. In such cases, the grains should be washed multiple times and soaked in lime water (pH 8–9) for 6 hours until the kernels are fully cooked. Afterward, they are rinsed with cold water and ready for use.
2. **Basic Formula**:
- Wheat grains (dry weight): 100 kg
- Cow dung powder: 30 kg
- Gypsum: 1 kg
- Calcium carbonate: 1.5 kg
3. **Substrate Mixing**: The wheat kernels are mixed with cow dung powder in a 1:0.2 ratio, along with other additives. It’s crucial that the cow dung is evenly coated on the grains. Any remaining cow dung powder is reserved for later use.
4. **Bottling Process**: For small-scale production, standard bottles are not always used. Instead, 500 ml infusion bottles are commonly used to reduce costs. A layer of cow dung is placed at the bottom of each bottle, approximately 1–1.5 cm thick. Then, the wheat grains are added up to 1 cm below the bottle’s neck. A small amount of cow dung is added on top, and the contents are flattened and compacted using an "L"-shaped tool before being sealed with a tampon.
5. **Sterilization and Cooling**: Autoclaving is used at 0.15 MPa for 2–2.5 hours. Once the pressure drops to zero, the bottles are moved to a sterilized cooling room. The room should be cleaned and disinfected in advance, and the bottles are inoculated once cooled to 30°C.
6. **Inoculation Procedure**: Before inoculation, the room is sprayed with Saibai 09 solution. The original culture is wiped with 75% alcohol, and after 10 minutes, the inoculation process begins. Each 500 ml bottle can receive over 40 inoculations.
7. **Cultivation Conditions**: The culture room must be clean, and the floor should be watered and sprayed with a disinfectant solution. After two days of confinement, lime powder is applied to the floor to transfer the inoculated strains. During the mycelium growth phase, the temperature should be kept around 25°C, not exceeding 28°C. If conditions are not ideal, the temperature can be lowered by placing the bottles in groundwater. Bricks are raised 20 cm above the ground, covered with plastic, and the bottles are submerged vertically. Water is changed 1–2 times daily to maintain a cool environment. A preventive spray with Silly Solution or Saibai 09 is applied every 3 days. If pests like fungus gnats appear, high-efficiency cypermethrin is used for control.
8. **Contamination Management**: On day 3 post-inoculation, check for contamination. Any bottles showing non-Agaricus mycelium should be removed immediately. For Streptomyces contamination, the bottles are carefully extracted and disposed of. General bacterial contamination can be treated by spraying and reusing the bottles after drying. Clostridium contamination requires isolation and proper disposal, such as burning or deep burial. Empty bottles must be scalded with hot water above 70°C before reuse.
9. **Temporary Storage**: After 35–45 days, the cultures will begin to produce spores. Unused strains should be stored at low temperatures, ideally between 2°C and 4°C.
10. **Quality Assessment**: Healthy Agaricus bisporus mycelium is thick, white, and grows in a fan-like pattern with abundant hyphae. The “2796†strain is semi-aerobic, with consistent and vigorous growth. No yellow or white bacteria should be present. If the mycelium appears thin, weak, or shows signs of discoloration, or if there are strange odors or pests, the culture is considered substandard. Abnormal growth patterns—such as uneven development, excessive dryness, or high temperatures—indicate poor quality and should not be used for planting.
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