Secondary fermentation preparation method of cultivation material for agaricus blazei murill and agaricus blazei murill cultivation method

Abstract

The invention discloses a secondary fermentation preparation method of a cultivation material for agaricus blazei murill and an agaricus blazei murill cultivation method. The method comprises the following steps: sequentially carrying out primary tunnel fermentation and secondary tunnel sealing fermentation on pretreated raw materials of the cultivation material to obtain the cultivation material; the method comprises the following steps: 1) primary tunnel fermentation: mixing soaked and drained corncobs, bagasse and cow dung to obtain a fermentation material, filling the fermentation material into a first primary fermentation tunnel for primary fermentation, and after the ninth day when the temperature of a material pile subjected to the primary fermentation reaches 70-80 DEG C, adding ammonium bicarbonate and calcium superphosphate after the temperature of the pile core is reduced; and 2) secondary tunnel sealing fermentation: filling the primary fermentation material into a secondary sealing fermentation tunnel, and respectively carrying out a temperature equalization period, a heating period, a pasteurization period, a cooling period, a temperature control period and a cooling discharging period to obtain the fermentation material. The preparation and production time of more than 5 tons of fermentation materials can be effectively shortened to 26 days, and meanwhile, after the fermentation materials are inoculated with agaricus blazei murill strains, the proportion of first-grade products of agaricus blazei murill dried products obtained through cultivation is high and can reach 90% or above.

Description

Technical Field

[0001] The present application relates to the technical field of Agaricus blazei Murrill cultivation, and in particular to a secondary fermentation preparation method for Agaricus blazei Murrill cultivation material and an Agaricus blazei Murrill cultivation method. Background Art

[0002] Agaricus blazei is a rare edible and medicinal mushroom. It belongs to the phylum Basidiomycota, order Agaricales, family Agariciaceae, and genus Agaricus. It is native to the United States, Brazil, and other regions. It was first discovered by American mycologist W.A. Murrill. Artificial cultivation began in Japan, where it was named "Agaricus blazei." In 1992, this species was introduced to my country, where it was successfully cultivated for the first time and subsequently spread throughout the country.

[0003] Agaricus blazei Murrill has a smooth, crisp, and tender mushroom body, offering an excellent texture, a pure, fresh, and fragrant almond aroma. It is nutritious, rich in protein, amino acids, and minerals, and boasts a high nutritional value. It is also rich in polysaccharides, active nucleic acids, and other active substances, which have therapeutic benefits for stroke, myocardial infarction, renal insufficiency, neuralgia, and cancer. Consequently, Agaricus blazei Murrill has become a favorite among consumers, and in recent years, it has been widely cultivated nationwide, with both the area and scale of cultivation continuously expanding, demonstrating enormous potential for development.

[0004] Currently, domestic cultivation of Agaricus blazei Murrill primarily relies on simple agricultural methods, characterized by rudimentary facilities, outdated technology, small-scale production, low yields, and poor quality. The formula and nutrient ratios are irrational, and operations are non-standard. The pre-fermentation (primary fermentation) of the compost is affected by external climate factors (temperature, rain, strong winds, etc.), resulting in uneven fermentation. Inadequate pasteurization and temperature maintenance during the post-fermentation (secondary fermentation) lead to poor compost maturity and severe contamination with bacteria and insect eggs, severely impacting yields. Furthermore, the production process is labor-intensive and labor costs are high.

[0005] Existing fermentation methods for Agaricus blazei Murrill cultivation primarily rely on steam and dry heat. Existing steam fermentation methods, however, require heat generated by simple boiling pots, resulting in low thermal energy utilization and cumbersome operation, making efficient and convenient operation impossible. While dry heat disinfection simplifies operation and improves sterilization efficiency, it doesn't allow wet steam to penetrate the fermentation medium, resulting in incomplete elimination of insect eggs and pathogens. This only accelerates heating, but fails to fully and thoroughly heat the fermentation medium.

[0006] CN202210695980.6 discloses an Agaricus blazei strain ZJJSR001 and a cultivation method thereof. After the strain is cultivated and harvested using the existing cultivation method and the fungus is dried, the proportion of first-class mushrooms obtained according to the GH / T 1463-2024 classification standard is low.

[0007] The information disclosed in the background section is only intended to enhance understanding of the overall background of the invention and should not be considered as an admission or any form of suggestion that the information constitutes the prior art already known to a person of ordinary skill in the art. Summary of the Invention

[0008] In response to the above-mentioned technical problems, the present application provides a secondary fermentation preparation method for cultivation material for Agaricus blazei Murrill and an Agaricus blazei Murrill cultivation method, which can effectively shorten the preparation and production time of more than 5 tons of fermentation material to 26 days. At the same time, after the fermentation material is inoculated with Agaricus blazei Murrill strains, the proportion of first-class dried Agaricus blazei Murrill products obtained by cultivation can reach as high as over 90%.

[0009] The present application provides a secondary fermentation preparation method for agaricus blazei murill cultivation material and an agaricus blazei murill cultivation method, comprising the following steps: pre-treated cultivation material raw materials are sequentially subjected to primary tunnel fermentation and secondary tunnel sealed fermentation to obtain the cultivation material;

[0010] 1) Primary tunnel fermentation: soaking and draining corn cobs, sugarcane bagasse and cow dung to obtain a fermentation material, and loading the fermentation material into the first primary fermentation tunnel for the first fermentation. When the temperature of the pile of the first fermentation reaches 70-80°C on the 9th day, when the temperature of the pile core drops, ammonium bicarbonate and superphosphate are added, stirred evenly, and transferred to the second primary fermentation tunnel for the first water replenishment. On the second day, the material is turned over and gypsum powder and compound fertilizer are added and stirred. After the material is transferred to the third primary fermentation tunnel, the second water replenishment is at least 1 hour later. On the 17th day after the transfer to the third primary fermentation tunnel, limestone is added and stirred evenly. The third water replenishment and ventilation are carried out. On the 24th day after the fermentation material enters the third primary fermentation tunnel, the fourth water replenishment is carried out to obtain a fermentation material.

[0011] The obtained primary fermentation material has a moisture content of 70% and a pH value of 8.2. It has a dark brown appearance, no obvious ammonia smell, no odor, and no sour odor. It contains no fecal matter and is not sticky. The corn cob particles are softened and no lumps are found when pressed. The sugarcane bagasse breaks when pulled hard, and one or two drops of water drip from between the fingers when the material is squeezed hard. The water inlet flow rate during the water replenishment process is 1.0 to 3.0 meters per second.

[0012] The procedure for the first water replenishment is: replenish water for 30 minutes on the second day after the material enters the second primary fermentation tunnel;

[0013] Add water for 30 minutes on the 5th day after the material enters the second fermentation tunnel;

[0014] On the 8th day after the material enters the second fermentation tunnel, add water for 50 minutes;

[0015] 2) Secondary sealed fermentation in a tunnel: The primary fermentation material is loaded into a secondary sealed fermentation tunnel and subjected to a temperature equalization period, a temperature rise period, a pasteurization period, a cooling period, a temperature control period, and a temperature drop and discharge period to obtain a fermentation material; the obtained fermentation material has a moisture content of 68% and a pH value of 7.5-8;

[0016] Temperature equalization period: hot air of 55-57℃ is introduced to raise the temperature of the pile to 48-50℃ within 12 hours;

[0017] Heating period: hot air at 60-63℃ is introduced to raise the temperature of the pile to 55-58℃ within 10-12 hours;

[0018] Pasteurization period: hot air at 60-63℃ is introduced to maintain the temperature of the pile at 55-58℃ for 8 hours;

[0019] Cooling period: The inlet air temperature is set at 40°C to reduce the fermentation temperature from 55-58°C to 44-46°C within 12-14 hours;

[0020] Temperature control period: maintain the fermentation material temperature at 45-48°C for 4-6 days;

[0021] Cooling and discharging period: reduce the material temperature to 24-26℃ within 4 hours;

[0022] The planting material is composed of 25-35 parts by weight of corn cobs, 30-40 parts by weight of bagasse, 3-8 parts by weight of cow dung, 2-3 parts by weight of ammonium bicarbonate, 2-3 parts by weight of superphosphate, 2-3 parts by weight of gypsum powder, 2-3 parts by weight of lime powder, and 0.5-1 part by weight of compound fertilizer;

[0023] The preservation certificate number of Agaricus blazei is CGMCC NO.40129, and its classification name is: Agaricus blazei.

[0024] Preferably, the compound fertilizer used has an effective calcium content of ≥1.0%, an effective magnesium content of ≥1.0%, a total sulfur content of ≥2.0%, and the contents of trace elements copper, iron, manganese, zinc, boron, and molybdenum are all ≥0.02%.

[0025] Preferably, the material pile height in the first primary fermentation tunnel, the second primary fermentation tunnel, and the third primary fermentation tunnel is 3 m and the width is 5.7 m.

[0026] Preferably, the first primary fermentation tunnel, the second primary fermentation tunnel, and the third primary fermentation tunnel are primary fermentation tunnels;

[0027] The primary fermentation tunnel includes: a tunnel area, multiple spray heads, a curved top cover, and a fan; one end of the tunnel area is closed and the other end is open; the top of the tunnel area is open; the top of the tunnel area is covered with a curved top cover; multiple spray heads are installed under the curved top cover; the spray heads are installed on the water inlet pipe, and the water inlet pipe is connected to the water storage tank;

[0028] An air outlet is provided on the closed end of the tunnel area, and the air outlet is connected to the air outlet pipeline of the fan.

[0029] Preferably, it comprises: a ventilation pipe; a plurality of grooves are arranged at intervals on the bottom surface of the tunnel area; a mezzanine is arranged at the bottom of the tunnel area, a plurality of air outlets are opened on the grooves, and the air outlets connect the mezzanine and the tunnel area;

[0030] A plurality of ventilation pipes are laid in the interlayer; one end of the ventilation pipe is connected with the air outlet pipe of the fan, and a plurality of through holes are arranged on the other end.

[0031] Preferably, the secondary sealed fermentation tunnel comprises: a sealed and insulated tunnel area, a sealed door, a low-pressure and high-air-volume system, multiple temperature and humidity sensors, multiple carbon dioxide and oxygen content detectors, and a touch-operated display; a feed port is provided at one end of the sealed and insulated tunnel area, and a sealed door is provided on the feed port for opening and closing; a low-pressure and high-air-volume system is provided on the outer side of the other end of the sealed and insulated tunnel area; an air outlet of the low-pressure and high-air-volume system is connected to a pipeline of the sealed and insulated tunnel area;

[0032] Temperature and humidity sensors, and carbon dioxide and oxygen content detectors are cross-spaced and arranged on the inner wall of the sealed and heat-insulated tunnel area;

[0033] The temperature and humidity sensors, the low-pressure and high-air-volume system, and the carbon dioxide and oxygen content detectors are electrically connected to the touch-operated display respectively.

[0034] Preferably, it comprises: a ventilation pipe; a plurality of grooves are arranged at intervals on the bottom surface of the sealed and heat-insulating tunnel area; a sandwich is arranged at the bottom of the sealed and heat-insulating tunnel area, a plurality of air outlets are opened on the grooves, and the air outlets are connected to the sandwich and the sealed and heat-insulating tunnel area;

[0035] A plurality of ventilation pipes are laid in the interlayer; one end of the ventilation pipe is connected with the air outlet pipe of the fan, and a plurality of through holes are arranged on the other end.

[0036] Preferably, during the secondary fermentation process, the oxygen concentration in the secondary sealed fermentation tunnel is controlled at 15% to 20%, and the carbon dioxide concentration in ppm is controlled at 400 to 800 ppm.

[0037] Another aspect of the present application provides a method for cultivating Agaricus blazei Murrill, comprising sowing Agaricus blazei Murrill wheat spawn at a sowing rate of 0.6 kg per square meter on the Agaricus blazei Murrill cultivation material obtained by the above method, and then incubating the mushrooms in a closed shed for one week before entering the mycelial germination stage. During the mycelial germination stage, the temperature of the mushroom house is controlled at 20°C to 25°C.

[0038] When the mycelium grows to 1 / 2 of the thickness of the material, loosen the material once with a rake. When the mycelium grows to 2 / 3 of the material layer, or when the material layer is fully eaten, cover the material with soil. Before covering the soil, spray water on the material surface several times to adjust the moisture. Cover the soil and then carry out soil covering management and mushroom fruiting management to obtain Agaricus blazei fruiting bodies and harvest.

[0039] Preferably, the soil covering management includes: spreading 1% quicklime on the soil, adjusting the soil pH to 7.0-7.5 and then stirring evenly, covering with plastic film and then stacking for one week, first covering with coarse soil with a thickness of 2 cm-3 cm and then spraying the soil particles with a sprayer;

[0040] When the hyphae climb up the soil layer and begin to twist, cover it with 0.5cm to 1.0cm thick fine soil. Within 2 days of covering the soil, moisten the soil with 1% lime water or clean water and keep the moisture content of the soil layer at 60%.

[0041] Fruiting management includes: When the fuzzy hyphae in the covering soil gradually transform into cords, some cords begin to kink and swell at their intersections, and white, millet-like primordia appear, spray thoroughly with water and increase ventilation after spraying. Thereafter, lightly spray the soil once or twice daily to keep it moist; ventilate the shed two or three times. Maintain the temperature in the shed at 20°C to 25°C and maintain indoor humidity at 85% to 90%. Adjust the diffuse light during the fruiting period until the mushrooms meet harvest standards.

[0042] The beneficial effects of this application include:

[0043] 1) The secondary fermentation preparation method for the cultivation material for Agaricus blazei provided in this application works together with the equipment and operating steps. Through secondary fermentation, after adding the raw materials in batches during the primary fermentation, multiple water replenishment is performed after adding ammonium bicarbonate and superphosphate to improve the penetration uniformity of the added raw materials in the pile. At the same time, combined with multiple water replenishment and batch addition during the primary fermentation process, a good post-fermentation effect is achieved for materials with different characteristics in the cultivation material. In addition, the temperature of the pile at each stage during the secondary fermentation process and the carbon dioxide and oxygen content during the secondary fermentation process are effectively controlled to improve the fermentation degree of raw materials with different characteristics in the obtained fermentation material. This is conducive to the full utilization of the fermentation material by the strain after subsequent colonization of the strain.

[0044] 2) The secondary fermentation method for preparing a culture medium for Agaricus blazei Murrill provided in this application further matures the culture medium after the second fermentation, promoting the growth and development of a large number of beneficial microorganisms, promoting nutrient decomposition and accumulation of effective nutrients, and killing pests and harmful microorganisms in the culture medium. This allows the Agaricus blazei Murrill strains to germinate faster, allowing the germinated mycelium to adapt to the nutritional environment in the shortest possible time and grow and develop smoothly, thereby increasing the proportion of first-grade Agaricus blazei Murrill obtained.

[0045] 3) The secondary fermentation method for preparing cultivation materials for Agaricus blazei Murrill provided in this application offers a shorter fermentation time of only 26 days compared to traditional pile fermentation. It also provides excellent fermentation quality, with a moisture content of 70% and a pH of 8.2. It is energy-efficient and uses natural microbial fermentation. It is highly efficient, with a single fermentation yield of up to 80 tons, making it suitable for large-scale industrial production. It integrates wastewater recycling technology, ensuring precise water replenishment during the primary tunnel and transfer period throughout the production process. Excess water flows back to the soaking tank through a drainage ditch and is reused for soaking the raw materials, eliminating wastewater discharge.

[0046] 4) The secondary fermentation method for preparing Agaricus blazei Murrill culture medium provided in this application takes only 32 days, from pre-wetting the raw materials to the final high-quality raw materials shipped, compared to the 45 days typically required for conventional culture medium fermentation. Microbial fermentation heat is utilized to complete the pasteurization and composting processes. Compared to the large-scale steam-consuming bed fermentation process, tunnel fermentation offers high throughput, low energy consumption, high efficiency, and no environmental pollution. The integrated primary and secondary tunnels provide centralized, efficient fermentation technology, ensuring stable and reliable quality and low production costs.

[0047] 5) The cultivation method of Agaricus blazei Murrill provided in this application, using the cultivation material obtained by the above method, and using this cultivation material in the cultivation of Agaricus blazei Murrill with a preservation certificate number of CGMCC NO.40129, can effectively increase the first-grade proportion of the obtained dried Agaricus blazei Murrill to more than 90%, while also increasing the yield. The Agaricus blazei ZJJSR001 used was deposited at the China General Microbiological Culture Collection Center (CGMCC) of the China Culture Collection Administration on April 18, 2022. The address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101. The deposit number is CGMCC NO.40129, and the recommended classification name is: Agaricus blazei. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 A schematic diagram of a secondary fermentation method for preparing a cultivation material for Agaricus blazei Murrill and a method for cultivating Agaricus blazei Murrill in at least one embodiment provided in the present application;

[0049] Figure 2 A schematic diagram of a main cross-sectional view of a primary fermentation tunnel in at least one embodiment provided in this application;

[0050] Figure 3 A schematic diagram of a side end view of the opening of a primary fermentation tunnel in at least one embodiment provided in the present application;

[0051] Figure 4A physical photo of the primary fermentation tunnel in at least one embodiment provided in this application;

[0052] Figure 5 A schematic diagram of a front and cross-sectional view of a secondary sealed fermentation tunnel in at least one embodiment provided in this application;

[0053] Figure 6 Photos of the secondary sealed fermentation tunnel in at least one embodiment provided in this application; a) is an end view; b) is the internal structure of the sealed and heat-insulating tunnel area;

[0054] Figure 7 A schematic diagram of the connection structure of the secondary sealed fermentation tunnel module in at least one embodiment provided in this application;

[0055] Figure 8 A schematic diagram of a partially enlarged structure of a ventilation duct in at least one embodiment provided in this application;

[0056] Legend:

[0057] First primary fermentation tunnel 131, second primary fermentation tunnel 132, third primary fermentation tunnel 133, tunnel area 32, spray head 22, arc-shaped top cover 321, fan 1, groove 231, ventilation pipe 23, through hole 232, sealed and heat-insulated tunnel area 211, sealed door 212, low-pressure and high-air-volume system 2, temperature and humidity sensor 13, carbon dioxide and oxygen content detector 141, touch-operated display 12, air inlet 11, and secondary sealed fermentation tunnel 14. DETAILED DESCRIPTION

[0058] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments, but the present invention is not limited in any way. Any changes or improvements made based on the teachings of the present invention fall within the scope of protection of the present invention.

[0059] The controller used in this embodiment is an existing structure, and the control circuit can be implemented through simple programming by technicians in this field. It is common knowledge in this field and is only used without modification. Therefore, the control method and circuit connection will not be described in detail.

[0060] Example

[0061] Unless otherwise specified, the materials and instruments used in the following examples were obtained from commercial channels; the detection methods used were all existing methods unless otherwise specified.

[0062] Example 1 Agaricus blazei cultivation material centralized secondary fermentation

[0063] 1. Raw materials

[0064] 1. The raw materials of the culture medium are mixed and prepared by mixing the following ingredients: 30 tons of corn cobs, 36 tons of bagasse, 5 tons of cow dung, 2.3 tons of ammonium bicarbonate, 2.3 tons of superphosphate, 2.3 tons of gypsum powder, 2.3 tons of lime powder, and 0.6 tons of compound fertilizer.

[0065] Corncobs: Corncobs are not limited to any length and must be crushed into granules with a diameter of 5-8 cm. They must be free of mold and have a moisture content of 15-18%.

[0066] Sugarcane bagasse: It needs to be crushed to remove large pieces of bagasse. It should be free of mold and have a moisture content of 15-18%.

[0067] Cow dung: The transported cow dung is placed in a storage tank for more than 15 days to allow it to dry naturally, with a moisture content of 80-85%.

[0068] Compound fertilizer: effective calcium ≥ 1.0%, effective magnesium ≥ 1.0%, total sulfur ≥ 2.0%, trace elements copper, iron, manganese, zinc, boron, molybdenum ≥ 0.02%

[0069] Ammonium bicarbonate: white or light-colored crystals, total nitrogen ≥17.1%, moisture content ≤7.2%

[0070] Superphosphate: available phosphorus ≥18.0%, water-soluble phosphorus ≥13.0%, free acid ≤5.5%, particle size ≥80%.

[0071] Gypsum powder: non-crystalline calcium sulfate, purity ≥98%, fineness ≥100 mesh, calcium content 20-28%, sulfur content 16-22%.

[0072] Lime powder: alkali content ≤0.4%, activity ≥80%, impurity content ≤8%, calcium content ≥75%.

[0073] 2. Pre-wetting of raw materials

[0074] Use a loader to mix 30 tons of corn cobs and 36 tons of sugarcane bagasse evenly and place them in a soaking tank for at least 24 hours. As the corn cobs and sugarcane bagasse absorb water, their texture begins to soften, which is beneficial for the subsequent fermentation process.

[0075] 3. Mixing of main raw materials

[0076] Remove the soaked corncobs and sugarcane bagasse from the soaking tank and drain for 3-5 hours to remove any excess water. Then, add cow dung and stir. Mix the cow dung evenly and stir until the cow dung, corncobs, and sugarcane bagasse are fully mixed, allowing the cow dung to fully absorb the water. This creates a fermentation material, which is then used in the subsequent fermentation.

[0077] 2. Primary Tunnel Fermentation

[0078] The fermentation material is loaded into the first fermentation tunnel for fermentation, and a large amount of heat is generated under the action of microorganisms. The temperature of the fermentation material is required to be controlled between 70 and 80°C during one fermentation. The operator regularly measures the temperature inside the pile core. When the temperature is too high, the blower 21 is turned on to ventilate the pile to achieve cooling. When the temperature of the fermentation material exceeds 500°C, the carbon source in the form of cellulose and hemicellulose in corn cobs and sugarcane bagasse is decomposed under the action of high-temperature thermophilic microorganisms.

[0079] On the 9th day after the start of the primary fermentation, the core temperature is measured every 2 hours. When the temperature here reaches 70-80°C and begins to drop, the ammonium bicarbonate and superphosphate weighed as above are added. After stirring evenly, a throwing machine is used to transfer the first fermentation material from the first primary fermentation tunnel 131 to the second primary fermentation tunnel 132.

[0080] The first fermentation material after the first tunnel fermentation is filled into the ventilation floor of the second primary fermentation tunnel 132 by a throwing machine, and is evenly spread to fill the tunnel. The filling width is 5.7m and the filling height is 3m.

[0081] Turn on the fan 1 to deliver fresh air to the fermentation material in the second primary fermentation tunnel 132 through the fan 1. Set the frequency converter to 42 Hz, ventilate for 5 minutes, and stop for 20 minutes.

[0082] On the second day after the first fermentation material enters the second fermentation tunnel 132, the water replenishing device is turned on and the water replenishing operation is carried out for 30 minutes; tap water is used as the inlet water, and the flow rate of the inlet water is 1.0 to 3.0 m / s. The inlet water passes through multiple sprinkler heads 22 arranged above the material pile to water the material pile. The following water replenishing operations are carried out in this way.

[0083] On the fifth day after the first fermentation material enters the second first fermentation tunnel 132, the water replenishing device is turned on and the water replenishing operation is performed for 30 minutes;

[0084] On the eighth day after the first fermentation material enters the second primary fermentation tunnel 132, the water replenishing device is turned on and the water replenishing operation is performed for 50 minutes;

[0085] On the 9th day after the first fermentation material enters the second fermentation tunnel 132, gypsum powder and compound fertilizer are added to the material for stirring and mixing to obtain the second fermentation material.

[0086] The obtained second fermentation material is transferred to the third primary fermentation tunnel 133, the fan parameters are set unchanged, and water is added for 1 hour;

[0087] On the 17th day after the second fermentation material enters the third primary fermentation tunnel 133, lime powder is added, mixed and stirred evenly, and water is added for 30 minutes; the frequency converter of fan 1 is set to 28, ventilation is carried out for 20 minutes, and stop for 15 minutes.

[0088] On the 24th day after the second fermentation material enters the third primary fermentation tunnel 133, the water replenishing device is turned on and the water replenishing operation is performed for 30 minutes;

[0089] On the 25th day after the second fermentation material enters the third primary fermentation tunnel 133, the primary fermentation is completed during the material turning and discharging to obtain the fermentation material.

[0090] After the third fermentation, the fermented material was tested for quality: 70% moisture, pH 8.2. Appearance quality criteria included: dark brown, no noticeable ammonia odor, no odor, and no sour or peculiar smell. It also contained no lumps of feces and was non-sticky. The corncob particles were softened and easily pressed. The sugarcane bagasse broke with a firm pull, and one or two drops of water dripped between the fingers when the material was squeezed.

[0091] See also Figures 1 to 8 The primary fermentation tunnels used are as follows: Figures 2-4 As shown, the tunnel area 32 measures 24m long x 5.7m wide x 4m high. It has an open sidewall at one end and an open top covered by a curved roof hood 321. A fan 1 is installed behind the fermentation tunnel. This fan 1 is connected to multiple ventilation pipes 23 installed in the ventilation floor. Through multiple through-holes 232 uniformly distributed throughout the ventilation pipes 23, it ventilates the piles of material laid within the tunnel area 32, achieving cooling and maintaining oxygen levels. The ventilation floor is provided with multiple grooves 231; the ventilation pipes 23 are installed in the floor below the grooves 231 and ventilate through the through-holes 232.

[0092] The ventilation pipe 23 used is a PVC pipe, and the longitudinal spacing of the through holes 232 is 300 mm. The ground and the wall are concrete reinforced structures.

[0093] 3. Secondary Tunnel Sealed Fermentation

[0094] See also Figures 5-7The secondary sealed fermentation tunnel 14 includes: a sealed and heat-insulating tunnel area 211, a sealed door 212, a low-pressure and high-air volume system 2, multiple temperature and humidity sensors 13, and multiple carbon dioxide and oxygen content detectors 141; the sealed and heat-insulating tunnel area 211 is a completely enclosed tunnel structure with heat-insulating function; an insulation layer structure is arranged on the outer wall of the sealed and heat-insulating tunnel area 211, and the top and closed end of the sealed and heat-insulating tunnel area 211 are both in a closed state, and a sealed door 212 is arranged on the other end, and the sealed door 212 is hinged to the sealed and heat-insulating tunnel area 211 for easy opening and closing. An air inlet 11 is provided at one end of the low-pressure, high-volume system 2. A touchscreen display 12 is provided on the outer wall of the low-pressure, high-volume system 2 for easy control. The touchscreen display 12 is electrically connected to multiple temperature and humidity sensors 13 and a carbon dioxide and oxygen content detector 141 provided on the inner wall of the sealed, insulated tunnel area 211, thereby enabling the temperature, humidity, carbon dioxide, and oxygen content of each area of the fermentation material within the sealed, insulated tunnel area 211 to be monitored. This allows the operator to perform various operations based on changes in temperature, humidity, oxygen, and carbon dioxide content within the sealed, insulated tunnel area 211. During the secondary fermentation process, the oxygen concentration is controlled at 15% to 20%, and the carbon dioxide concentration is controlled at 400 to 800 ppm.

[0095] A low-pressure, high-volume air system 2 is provided on the outside of the closed end of the sealed and insulated tunnel area 211, which can accurately control the material temperature. The most important pasteurization stage and the cultivation stage of beneficial microorganisms in the fermentation are both completed in the second stage of fermentation. The use of this fermentation method and equipment is particularly suitable for the aforementioned culture material combination.

[0096] The secondary fermentation process is controlled according to the following control process. The control system computer automatically controls the fermentation material temperature according to the curve, which can be unmanned. The entire secondary fermentation time is 6 to 8 days.

[0097] ① Temperature Equalization Phase: Maintained for 12 hours from the start of fermentation, the temperature at each measurement point is read. The fermentation material temperature range is 48-50°C. When the pile temperature measured by the temperature and humidity sensor 13 is below this temperature range, the inlet temperature of the low-pressure, high-volume air system 2 is 55-57°C. This phase primarily involves the operation of the low-pressure, high-volume air system 2 to achieve temperature uniformity across the fermentation material, laying the foundation for the subsequent temperature increase phase.

[0098] ② Heating period: When the temperature of the pile is raised to 55-58°C within 10-12 hours, the inlet temperature of the low-pressure and high-volume system 2 is raised to 60-63°C. The fresh air volume in this stage is reduced to 2 / 3 of the original inlet air volume, so that the fermentation temperature increases.

[0099] ③ Pasteurization period: The air temperature is 60-63°C to maintain the fermentation temperature at 55-58°C and maintain this temperature for 8 hours. This stage is one of the most important stages in the fermentation process and can effectively kill some bacteria and insect eggs.

[0100] ④ Cooling period: The inlet air temperature is set at 40°C to reduce the fermentation material temperature from 55-58°C to 44-46°C within 12-14 hours. During this stage, the control system increases the fan frequency and increases the fresh air volume to the original inlet air volume to achieve the purpose of cooling.

[0101] ⑤ Temperature Control Period: The processing time is 4 to 6 days. The fermentation temperature is 45 to 48°C, and the inlet air temperature is set at 49 to 53°C. During this stage, the control system keeps the temperature of the fermentation medium stable at 45 to 48°C. At a constant temperature of 46°C, actinomycetes multiply and decompose the culture medium. This is one of the key stages that affects the quality of the fermentation medium.

[0102] ⑥ Cooling and discharging period: The processing time is 4 hours, and the temperature of the fermented material drops from 45-48℃ to 24-26℃. When the temperature of the fermented material drops to 24-26℃, it can be discharged and the Agaricus blazei Murrill can be sown.

[0103] The secondary sealed fermentation tunnel has a length x width x height of 24mx4mx4m. A filtration and fresh air system is installed on the top of the fermentation tunnel ( Figure 2 ).

[0104] The sealed and insulated tunnel area 211 has multiple grooves 231 formed on its inner bottom surface. A ventilation duct 23 is installed beneath these grooves. Multiple through-holes 232 are spaced apart in the ventilation duct 23, and multiple exhaust holes are located at the bottom of the grooves 231. Air passes through the through-holes 232 and into the exhaust holes, eventually entering the sealed and insulated tunnel area 211. One end of the ventilation duct 23 is sealed, while the other end is connected to the low-pressure, high-volume airflow system 2. An outlet for the low-pressure, high-volume airflow system 2 is located above the closed end of the sealed and insulated tunnel area 211, allowing for comprehensive ventilation of the fermentation material within the sealed and insulated tunnel area 211. Air enters the sealed and insulated tunnel area 211 through the upper outlet and simultaneously enters all parts of the fermentation material through the through-holes 232 in the ventilation floor, ensuring comprehensive ventilation of the fermentation material. The ventilation duct 23 is made of PVC pipe, with the through-holes 232 spaced 200 mm apart. The floor of the sealed and insulated tunnel area 211 is a concrete pipe reinforced concrete structure. The exterior wall panels are constructed of an aluminum-zinc-coated substrate with an internal foam insulation layer.

[0105] After secondary fermentation, the technical specifications for the fermentation material are: 68% moisture content and a pH of 7.5-8. Because the secondary fermentation is pasteurized, the fermentation material used for Agaricus blazei Murrill cultivation is a very pure carrier. Therefore, strict disinfection measures must be implemented in the secondary fermentation discharge area. The ground in the discharge area, and vehicles and personnel entering the discharge area must be sprayed with Lysol disinfectant to prevent contamination of the fermentation material with foreign bacteria.

[0106] The fermentation medium emerging from the secondary tunnel is the culture medium needed for production and can be used directly for inoculation. This is significantly different from culture medium produced by traditional methods. After the secondary tunnel's centralized fermentation, the culture medium contains abundant actinomycetes, is gray-brown in color, non-sticky, and elastic. The extract is transparent, has a pH of 7.5-8, a water content of 68%, and no ammonia odor.

[0107] Example 2 Secondary concentrated fermentation cultivation material Agaricus blazei

[0108] In this example, Agaricus blazei Murrill was sown by broadcasting. When the fermentation medium temperature was stable at 24-26°C, Agaricus blazei Murrill spawn was broadcasted into the fermentation medium obtained in Example 1. The inoculum size was 0.6 kg per square meter. To sow, the spawn was first loosened and then evenly broadcast over the surface of the culture medium. After sowing, the surface was flattened and compacted with a wooden board or the palm of your hand.

[0109] 1. Management of fungal culture

[0110] After sowing, close the doors and windows and keep the spawning house closed for a week. Control the humidity and ventilation of the mushroom house to achieve the optimal conditions for mycelial germination and growth. During the mycelial germination period, the temperature in the mushroom house should be kept between 20℃ and 25℃. Ventilate the cultivation house for one hour a day. The humidity should be between 50% and 60%. Mycelial germination will begin 2 days after sowing, and the mycelium will cover the surface of the material on the 7th day.

[0111] Ten days after sowing, when the mycelium has grown to half the thickness of the compost, loosen the compost with a rake to increase ventilation and encourage rapid mycelial growth to the lower layers. 25-30 days after sowing, when the mycelium has fully grown, cover the compost with soil when it has grown to two-thirds of the compost layer, or has completely consumed the entire layer. Cover the compost with soil promptly. Before covering, spray the compost surface with water several times to adjust the moisture level. This will allow the mycelium to quickly rise to the surface.

[0112] 2. Soil cover management

[0113] When covering the soil, use a mixture of coarse and fine soil. Before use, sprinkle 1% quicklime on the soil to be used, mix evenly, cover with plastic film, and pile it for a week. Use lime water to adjust the soil pH to 7.0-7.5 before use.

[0114] When covering the soil, start with coarse soil. Spread the treated coarse soil evenly over the surface to a thickness of 2-3 cm. Use a sprayer to wet the soil particles until there are no white cores and the soil can be squeezed flat without being sticky. Once the mycelium has climbed up the soil layer and begun to tangle (generally after 8-12 days of covering the coarse soil), cover with fine soil to a thickness of 0.5-1.0 cm, ideally filling the gaps between the coarse soil particles. Within 2 days of covering the soil, moisten the soil several times with 1% limewater or clean water. The principle is to spray sparingly and frequently to keep the soil moist, with a moisture content of 60%.

[0115] 3. Mushroom production management

[0116] Under normal circumstances, 15 to 20 days after covering with soil, when the fuzzy hyphae in the covering layer gradually transform into cords, some cords begin to kink and swell at their intersections, and white, millet-like primordia appear, spray thoroughly with water. Spray evenly and gently, and increase ventilation after spraying. Thereafter, lightly spray water once or twice daily to keep the soil moist and free of white cores. Ventilate the shed two or three times, maintaining indoor humidity at 85% to 90%, and controlling the temperature at 20°C to 25°C. Diffuse light is required during the fruiting period; excessive darkness or intense light is detrimental to growth.

[0117] The principle of watering is to water thoroughly once for each crop of mushrooms and spray lightly several times. Spray more on sunny days, less on cloudy days, and stop spraying on rainy days. When the white granular mushroom buds on the mushroom bed grow to a diameter of 2cm to 3cm, stop spraying.

[0118] After each crop of mushrooms is harvested, the soil should be filled in time to keep the bed level, and the remaining mushroom stems and dead mushrooms should be removed in time. Water should be stopped for 3 to 4 days, and then heavy water should be sprayed to promote mushroom production. The entire fruiting period can last for 2 to 3 months, and generally 5 to 8 crops can be harvested.

[0119] 4. Harvesting and processing

[0120] It takes 5 to 7 days for the bud to mature into a mature mushroom. Harvest when the fruiting body of Agaricus blazei is 5 to 8 cm tall, the cap is light brown with fibrous scales, the cap is 3.5 to 5 cm in diameter, and the stipe is 4 to 7 cm long. It should not have opened or broken the mycelium. Harvesting can be done 2 to 3 times a day in warm weather, and once or twice a day in cool weather.

[0121] Comparative Example 1

[0122] The difference from Example 2 is that the fermentation material inoculated was a compost pile grown outdoors for 45 to 60 days, turned every 7 days, and fermented once. The amount of actinomycetes in the fermentation material and the condition of the mycelial growth in the culture medium showed that the conventionally produced culture medium had fewer actinomycetes and dense hairy mycelium, indicating that the culture medium had a low degree of decomposition, making it difficult for the mycelial cells to effectively utilize the nutrients contained therein, and the quality of the culture medium was poor.

[0123] After harvesting the fruiting bodies obtained in Example 2 and Comparative Example 1, dried Agaricus blazei Murrill products were prepared according to GH / T 1463-2024. The first-grade product quality (first-grade product quality / total yield) and the proportion of second-grade products (second-grade product quality / total yield) of the resulting dried products were determined according to GH / T 1463-2024. The fruiting time, yield, and proportion of high-quality mushrooms (high-quality mushroom quality / total yield) for Example 2 and Comparative Example 1 were also statistically analyzed and listed in Table 1.

[0124] Table 1 Comparison between traditional fermentation materials and centralized secondary fermentation materials

[0125]

[0126]

[0127] As can be seen from Table 1, the fermentation material obtained by the method provided in this application can be used to ferment Agaricus blazei Murrill, which can effectively increase the proportion of first-class products in the obtained dried Agaricus blazei Murrill products, improve product quality and yield, reduce losses, and shorten the fruiting time.

[0128] Example 3

[0129] The difference from Example 1 is that the composition of the planting material is 25 parts by weight of corn cobs, 30 parts by weight of bagasse, 3 parts by weight of cow dung, 2 parts by weight of ammonium bicarbonate, 2 parts by weight of superphosphate, 2 parts by weight of gypsum powder, 2 parts by weight of lime powder, and 0.5 parts by weight of compound fertilizer.

[0130] Example 4

[0131] The difference from Example 1 is that the composition of the planting material is 35 parts by weight of corn cobs, 40 parts by weight of bagasse, 8 parts by weight of cow dung, 3 parts by weight of ammonium bicarbonate, 3 parts by weight of superphosphate, 3 parts by weight of gypsum powder, 3 parts by weight of lime powder, and 1 part by weight of compound fertilizer.

[0132] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A secondary fermentation method for preparing a cultivation material for Agaricus blazei Murrill, characterized in that: The following steps are involved: The pre-treated cultivation material raw materials are sequentially subjected to a primary tunnel fermentation and a secondary tunnel sealed fermentation to obtain the cultivation material; 1) Primary tunnel fermentation: corn cobs, bagasse and cow dung that have been soaked and drained are mixed to obtain fermentation material, and the fermentation material is loaded into the first primary fermentation tunnel (131) for the first fermentation. When the temperature of the pile of the first fermentation reaches 70-80°C on the 9th day, ammonium bicarbonate and superphosphate are added after the pile core temperature drops, and the mixture is evenly stirred. The mixture is transferred to the second primary fermentation tunnel (132) for the first water replenishment. On the second day, the mixture is turned over and gypsum powder and compound fertilizer are added and stirred. The mixture is transferred to the third primary fermentation tunnel (133). After the second water replenishment for at least 1 hour, limestone is added on the 17th day after the mixture is transferred to the third primary fermentation tunnel (133) and the mixture is evenly stirred. The mixture is watered for the third time and ventilated. The fermentation material is watered for the fourth time on the 24th day after entering the third primary fermentation tunnel (133) to obtain the primary fermentation material. The obtained primary fermentation material has a moisture content of 70% and a pH value of 8.

2. It has a dark brown appearance, no obvious ammonia smell, no odor, and no sour odor. It contains no fecal matter and is not sticky. The corn cob particles are softened and no lumps are found when pressed. The sugarcane bagasse breaks when pulled hard, and one or two drops of water drip from between the fingers when the material is squeezed hard. The water inlet flow rate during the water replenishment process is 1.0 to 3.0 meters per second. The procedure for the first water replenishment is: replenish water for 30 minutes on the second day after the material enters the second primary fermentation tunnel (132); On the fifth day after the material enters the second primary fermentation tunnel (132), water is added for 30 minutes; On the 8th day after the material enters the second primary fermentation tunnel (132), water is added for 50 minutes; 2) Secondary tunnel sealed fermentation: After the primary fermentation material is loaded into the secondary sealed fermentation tunnel (14), it undergoes a temperature equalization period, a temperature rise period, a pasteurization period, a cooling period, a temperature control period, and a temperature drop and discharge period to obtain a fermentation material; the obtained fermentation material has a water content of 68% and a pH value of 7.5-8; Temperature equalization period: hot air of 55-57℃ is introduced to raise the temperature of the pile to 48-50℃ within 12 hours; Heating period: hot air at 60-63℃ is introduced to raise the temperature of the pile to 55-58℃ within 10-12 hours; Pasteurization period: hot air at 60-63℃ is introduced to maintain the temperature of the pile at 55-58℃ for 8 hours; Cooling period: The inlet air temperature is set at 40°C to reduce the fermentation temperature from 55-58°C to 44-46°C within 12-14 hours; Temperature control period: maintain the fermentation material temperature at 45-48°C for 4-6 days; Cooling and discharging period: reduce the material temperature to 24-26℃ within 4 hours; The planting material is composed of 25-35 parts by weight of corn cobs, 30-40 parts by weight of bagasse, 3-8 parts by weight of cow dung, 2-3 parts by weight of ammonium bicarbonate, 2-3 parts by weight of superphosphate, 2-3 parts by weight of gypsum powder, 2-3 parts by weight of lime powder, and 0.5-1 part by weight of compound fertilizer; The preservation certificate number of Agaricus blazei is CGMCC NO.40129, and its classification name is: Agaricus blazei.

2. The method for preparing a cultivation material for Agaricus blazei Murrill by secondary fermentation according to claim 1, wherein: The effective calcium content of the compound fertilizer used is ≥1.0%, the effective magnesium content is ≥1.0%, the total sulfur content is ≥2.0%, and the contents of trace elements copper, iron, manganese, zinc, boron and molybdenum are all ≥0.02%.

3. The method for preparing a cultivation material for Agaricus blazei by secondary fermentation according to claim 1, wherein: The material piles in the first primary fermentation tunnel (131), the second primary fermentation tunnel (132), and the third primary fermentation tunnel (133) have a height of 3 m and a width of 5.7 m.

4. The method for preparing a cultivation material for Agaricus blazei Murrill by secondary fermentation according to claim 1, wherein: The first primary fermentation tunnel (131), the second primary fermentation tunnel (132), and the third primary fermentation tunnel (133) are primary fermentation tunnels; The primary fermentation tunnel comprises: a tunnel area (32), a plurality of spray heads (22), an arc-shaped top cover (321), and a fan (1); one end of the tunnel area (32) is closed and the other end is open; the top of the tunnel area (32) is open; the arc-shaped top cover (321) is provided on the open top of the tunnel area (32); a plurality of spray heads (22) are provided below the arc-shaped top cover (321); the spray heads (22) are installed on a water inlet pipe, and the water inlet pipe is connected to a water storage tank; An air outlet is provided on the closed end of the tunnel area (32), and the air outlet is communicated with the air outlet pipeline of the fan (1).

5. The method for preparing a cultivation material for Agaricus blazei Murrill by secondary fermentation according to claim 4, characterized in that: include: A ventilation pipe (23); a plurality of grooves (231) are arranged at intervals on the bottom surface of the tunnel area (32); a sandwich is arranged at the bottom of the tunnel area (32); a plurality of air outlets are opened on the grooves (231), and the air outlets communicate with the sandwich and the tunnel area (32); A plurality of ventilation pipes (23) are laid in the interlayer; one end of the ventilation pipe (23) is connected to the air outlet pipe of the fan (1), and a plurality of through holes (232) are provided on the other end.

6. The method for preparing a cultivation material for Agaricus blazei Murrill by secondary fermentation according to claim 1, characterized in that: The secondary sealed fermentation tunnel (14) comprises: a sealed heat preservation tunnel area (211), a sealed door (212), a low-pressure and high-air-volume system (2), a plurality of temperature and humidity sensors (13), a plurality of carbon dioxide and oxygen content detectors (141), and a touch-operated display (12); a feed port is provided at one end of the sealed heat preservation tunnel area (211), and a sealed door (212) is provided on the feed port for opening and closing; a low-pressure and high-air-volume system (2) is provided on the outer side of the other end of the sealed heat preservation tunnel area (211); an air outlet of the low-pressure and high-air-volume system (2) is connected to a pipeline of the sealed heat preservation tunnel area (211); The temperature and humidity sensors (13) and the carbon dioxide and oxygen content detectors (141) are arranged in a cross-spaced manner on the inner wall of the sealed and heat-insulating tunnel area (211); The temperature and humidity sensor (13), the low-pressure and high-air volume system (2), and the carbon dioxide and oxygen content detector (141) are electrically connected to the touch-operated display (12) respectively.

7. The method for preparing a cultivation material for Agaricus blazei Murrill by secondary fermentation according to claim 6, characterized in that: include: ventilation pipe (23); A plurality of grooves (231) are arranged at intervals on the bottom surface of the sealed and heat-insulating tunnel area (211); An interlayer is provided at the bottom of the sealed and heat-insulating tunnel area (211), and a plurality of air outlets are provided on the groove (231), wherein the air outlets communicate with the interlayer and the sealed and heat-insulating tunnel area (211); A plurality of ventilation pipes (23) are laid in the interlayer; one end of the ventilation pipe (23) is connected to the air outlet pipe of the fan (1), and a plurality of through holes (232) are provided on the other end.

8. The method for preparing a cultivation material for Agaricus blazei Murrill by secondary fermentation according to claim 1, characterized in that: During the secondary fermentation process, the oxygen concentration in the secondary sealed fermentation tunnel (14) is controlled at 15% to 20%, and the carbon dioxide concentration in ppm is controlled at 400 to 800 ppm.

9. A method for cultivating Agaricus blazei, characterized in that: Sowing the wheat spawn of Agaricus blazei Murrill at a sowing rate of 0.6 kg per square meter on the cultivation material for Agaricus blazei Murrill obtained by the method according to any one of claims 1 to 8, and then entering the mycelial germination period after a week of germination in a closed shed. The temperature of the mushroom house during the mycelial germination period is controlled at 20° C. to 25° C.; When the mycelium grows to 1 / 2 of the thickness of the material, loosen the material once with a rake. When the mycelium grows to 2 / 3 of the material layer, or when the material layer is fully eaten, cover the material with soil. Before covering the soil, spray water on the material surface several times to adjust the moisture. Cover the soil and then carry out soil covering management and mushroom fruiting management to obtain Agaricus blazei fruiting bodies and harvest.

10. The Agaricus blazei Murrill cultivation material method according to claim 9, characterized in that: Soil cover management includes: spreading 1% quicklime on the soil, adjusting the soil pH to 7.0-7.5 and stirring evenly, covering with plastic film and piled for a week, then covering with coarse soil with a thickness of 2-3 cm and then spraying the soil particles with a sprayer to wet them; When the hyphae climb up the soil layer and begin to twist, cover it with 0.5cm to 1.0cm thick fine soil. Within 2 days of covering the soil, moisten the soil with 1% lime water or clean water and keep the moisture content of the soil layer at 60%. Fruiting management includes: When the fuzzy hyphae in the covering soil gradually transform into cords, some cords begin to kink and swell at their intersections, and white, millet-like primordia appear, spray thoroughly with water and increase ventilation after spraying. Thereafter, lightly spray the soil once or twice daily to keep it moist; ventilate the shed two or three times. Maintain the temperature in the shed at 20°C to 25°C and maintain indoor humidity at 85% to 90%. Adjust the diffuse light during the fruiting period until the mushrooms meet harvest standards.

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