Anti-pollution inoculation method for edible mushroom sticks

Abstract

The invention discloses an edible mushroom stick anti-pollution inoculation method, which comprises the following steps: S00: firstly, carrying out primary disinfection on mushroom sticks by utilizing normal pressure sterilization (keeping at 100 DEG C for 24 hours), and transporting the mushroom sticks to a mushroom shed under a covering condition; s10, stacking in a fungus shed to form a fungus stick pile, and placing an aerial fog disinfectant in the closed fungus pile for fumigation and disinfection; s20, transferring the fumigated mushroom sticks to an inoculation box under a closed condition; s30, through the operation hole, the automatic punching device is used for punching the mushroom sticks in the inoculation box, then the strain inoculation device is used for inoculating strains into the hole, and an outer tying opening is arranged in a sleeving mode; s40, moving the inoculated and sealed mushroom sticks out of the inoculation box under a closed condition, and transferring the mushroom sticks to a mushroom cultivation greenhouse; anti-pollution measures are taken in the whole process of stick manufacturing, sterilization, stacking and inoculation, and the risk of infectious microbe invasion is systematically reduced; compared with natural cooling (especially #-shaped stacking), the mushroom stick pollution rate in the inoculation process can be effectively reduced.

Description

Technical Field

[0001] This invention relates to the field of mushroom substrate inoculation technology, and in particular to a method for preventing contamination during inoculation of edible mushroom substrates. Background Technology

[0002] Black fungus spawn bags are cylindrical culture media used for the artificial cultivation of black fungus. They serve as a carrier for the growth of black fungus mycelium and the emergence of fruiting bodies. Their main components are agricultural by-products such as sawdust and wheat bran. After being mixed with water, bagged, and sterilized at high temperature, black fungus spawn is inoculated. The mycelium grows and spreads inside the bag, absorbing nutrients, and eventually, under suitable conditions, black fungus grows from the bag. Making spawn bags is an important step in achieving large-scale artificial cultivation of black fungus. Then, a device similar to the sterile inoculation box for spawn bags disclosed in patent document CN213847982U can be used to inoculate the spawn bags.

[0003] During the preparation of black fungus spawn, southern my country experiences a hot and humid season, which is highly conducive to the growth and reproduction of microorganisms. However, the spawn bags are packed tightly, and the substrate is primarily composed of coarse sawdust with numerous thorns, easily puncturing the bags and creating pinholes. These pinholes become entry points for contaminating the spawn. Since the optimal inoculation period for black fungus spawn is very short—generally only about 15 days at the same altitude—production companies need to produce large quantities of spawn in a short time to meet farmers' demands. Under these conditions and demands, the risk of contamination increases significantly if the spawn is not properly controlled. Currently, the crisscross stacking method is commonly used after spawn preparation, which has poor resistance to contamination. Once the spawn is contaminated, it not only affects the normal growth of the edible fungi, leading to reduced yield and poor quality, but in severe cases, it can even cause the entire batch of spawn to be scrapped, resulting in huge losses for producers and farmers. Therefore, a method for spawn inoculation that can effectively control contamination throughout the entire process of black fungus spawn stacking and inoculation is needed. Summary of the Invention

[0004] In order to overcome the shortcomings of the prior art, the technical problem to be solved by the present invention is to propose a method for preventing contamination during the inoculation of edible fungus spawn. Through a whole-process control system from spawn preparation, sterilization, stacking to inoculation, the contamination rate of black fungus spawn can be significantly reduced.

[0005] To achieve this objective, the present invention adopts the following technical solution: The present invention provides a method for preventing contamination during inoculation of edible mushroom substrate, comprising the following steps: S00: The mushroom sticks are first sterilized at 100℃ for 24 hours under normal pressure for preliminary disinfection. The sterilized mushroom sticks are then removed from the sterilization furnace while still hot at a temperature not lower than 85℃ and transported to the mushroom shed under covered conditions. S10: A first sealing film is laid on the ground inside the mushroom shed. Several rows of first mushroom sticks are stacked on the first sealing film in a linear arrangement. Based on the first mushroom stick layers, several rows of mushroom stick piles are stacked up one by one. The row spacing A between two adjacent rows of mushroom stick piles is 1 to 1.5 times the length L of the mushroom sticks. The height of the mushroom stick pile includes 9 to 12 layers of mushroom sticks. A second sealing film is laid on top of the several rows of mushroom stick piles. The periphery of the first sealing film and the second sealing film are sealed to form a closed mushroom pile. An aerosol disinfectant is placed in a container and placed between two adjacent rows of mushroom stick piles for fumigation disinfection. The amount of aerosol disinfectant used is 1 to 4 kg per 10,000 mushroom sticks. S20: The fumigated and disinfected mushroom logs are transferred to the inoculation box under sealed conditions. The steps of transferring the mushroom logs to the inoculation box are as follows: the transfer is carried out through a transfer chamber equipped with a disinfection function; the outer door of the transfer chamber is opened, the mushroom logs are placed in, and then closed. Subsequently, aerosol and / or ultraviolet disinfection is carried out in the chamber. After disinfection, the inner door is opened and the mushroom logs are sent into the inoculation box. The inside of the inoculation box is a sterile environment that has been sterilized. The inoculation box is equipped with at least one operating hole with an operating sleeve, an automatic punching device, and a mushroom inoculation device. S30: Through the operating hole, the automatic punching device punches holes in the inoculation box, then the inoculation device inoculates the inoculation into the holes, and the inoculation holes are sealed by covering them with a mushroom bag. S40: After inoculation and sealing, remove the inoculation box under sealed conditions and transfer it to the mushroom cultivation shed.

[0006] In step S20, the automatic punching device includes a punching head driven by a servo motor, and the punching depth, hole diameter, and hole spacing are automatically adjusted according to the length and diameter of the mushroom stick; wherein, the punching depth H satisfies:

[0007] in, The diameter of the mushroom log; To optimize the constant, the range is 1~2 cm; The aperture ranges from 3.6 to 4 cm; The hole spacing S satisfies:

[0008] Where L is the length of the mushroom log; E is the end distance, which is the distance between the holes at the outermost two ends and the two ends of the mushroom log, which is 3~5 cm; and N is the number of holes.

[0009] In step S20, the inoculation device is integrated inside the automatic punching device, and the inoculum is injected into the bottom of the hole after punching is completed; the punching device also includes a visual positioning sensor for identifying the position of the mushroom stick; the box is also equipped with a humidity sensor, whose data is connected to an external controller for display and alarm.

[0010] In step S30, the inoculation box is provided with two oppositely arranged operating holes and corresponding operating sleeves, forming a two-person collaborative workstation; the top of the box is provided with a transparent observation window and a lighting lamp, the bottom of the box is provided with casters, and the side of the box is provided with a closed conveyor pipe interface for outputting the inoculated mushroom sticks. It also includes a closed conveyor belt or mushroom cultivation cart that cooperates with the conveyor pipe interface, for transferring the inoculated mushroom sticks in a closed manner to the mushroom cultivation shed.

[0011] In step S20, the inoculation box is also equipped with a sensing control unit. After the mushroom stick is placed in the inoculation box, the sensing control unit automatically identifies the specifications of the mushroom stick and, based on the specifications and the mycelial growth model preset in the sensing control unit, automatically calculates and outputs the optimal number of inoculation holes, spatial layout, and depth information; drives the automatic punching device and the mycelial inoculation device connected to the control unit to perform synchronous punching and inoculation operations on the mushroom stick according to the output information.

[0012] The process of automatically calculating the optimal inoculation parameters based on specification parameters and a preset mycelial growth model includes: S21: Obtain the effective length (L) and diameter (D) of the mushroom log; S22: Based on the radial average growth rate of the mycelium of the target strain in the culture medium. Compared with the average longitudinal growth rate , and the preset time T for complete occupation of the target hyphae; S23: Calculate the number of inoculation points required for uniform distribution across the cross-section. Among them, the theoretical distance from any point on the cross-section to the nearest inoculation point is less than ,in, ; S24: Calculate the required number of inoculation rows in the longitudinal direction. Where the row spacing A satisfies: ; S25: Outputs the total number of inoculation wells N. .

[0013] The beneficial effects of this invention are as follows: (1) In this case, anti-contamination measures were implemented throughout the entire process from stick preparation, sterilization, stacking to inoculation, systematically reducing the risk of contamination by miscellaneous bacteria. Specifically, high-temperature isolation and fumigation disinfection were adopted in the sterilization and disinfection stage, that is, normal pressure sterilization (100℃ for 24 hours) combined with high temperature above 85℃ for cooking and covered transportation, which effectively killed pathogens. Then, sealed stacking (in a straight line) combined with fumigation with aerosol disinfectant significantly reduced the number of miscellaneous bacteria. Compared with natural cooling (especially well-shaped stacking), the contamination rate of the mushroom sticks can be effectively reduced. (2) During the sterilization process, a double-layer sealing film (plastic sheet) is used to isolate the outside air and maintain the high temperature and cleanliness inside the mushroom pile, that is, the sealed stacking creates a sterile environment; a protective film is also formed on the surface of the mushroom bag by aerosol disinfectant to reduce the risk of exposure and contamination during handling and transportation. (3) In addition to manual operation, the inoculation process can be further automated and precise. Inside the inoculation box, the automatic punching device is equipped with visual positioning and active drive to ensure that the punching depth, hole diameter and hole spacing are consistent. At the same time, the inoculation of the fungus and the punching are completed simultaneously. Then the fungus bag is put on to prevent subsequent contamination. Moreover, the mycelial growth model can be further added. The optimal punching depth and layout are automatically calculated according to the size of the fungus stick, so that the fungus is inoculated into the core area of ​​the fungus stick, so that the mycelium grows evenly and quickly. (4) By controlling pollution throughout the entire process and automating and intelligentizing the inoculation process, waste rods and repetitive labor caused by pollution can be effectively reduced, thereby reducing production costs and losses for farmers. Attached Figure Description

[0014] Figure 1 This is a schematic diagram illustrating the principle and flow of a method for preventing contamination during inoculation of edible mushroom substrate provided in a specific embodiment of the present invention; Figure 2 This is a schematic diagram of the mushroom sticks being stacked (in a straight line) in a method for preventing contamination of edible mushroom sticks provided in a specific embodiment of the present invention. Figure 3 This is a schematic diagram of the traditional method of stacking mushroom sticks (in a grid shape) according to a specific embodiment of the present invention; Figure 4 This is a schematic diagram illustrating the principle of a method for preventing contamination during inoculation of edible mushroom substrate in a specific embodiment of the present invention, where the substrate is stacked (in a straight line).

[0015] In the picture: 1. Mushroom logs; 2. First layer of mushroom logs; 3. Mushroom log pile; 4. First sealing film; 5. Second sealing film; 6. Container; 7. Mushroom shed; Detailed Implementation The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0016] In order to control contamination of the manufactured mushroom substrate in the early stage, reduce the risk of contamination, ensure the normal growth of edible fungi after inoculation, improve their yield and quality, and reduce losses for producers and farmers, this invention provides a method for preventing contamination inoculation of edible fungi substrate, based on the process of pretreatment and inoculation of mushroom substrate.

[0017] Example 1: This case focuses on "contamination prevention" and constructs a complete process control system from rod preparation, sterilization, stacking to inoculation, including the following steps: First, the prepared mushroom sticks 1 are sterilized after being covered with mushroom bags, i.e., step S00: the mushroom sticks 1 are first sterilized at 100℃ for 24 hours under normal pressure for preliminary sterilization. After sterilization, multiple mushroom sticks 1 are removed from the sterilization furnace while still hot at a temperature not lower than 85℃ and transported to the mushroom shed 7 under covered conditions. Specifically, the principle used here is pasteurization, which is to maintain a temperature of 62-65℃ for 30 minutes to kill various types of pathogenic bacteria, with a sterilization efficiency of 97.3%-99.9%. Maintaining a high temperature for a long time is an effective method derived from pasteurization to control the contamination of mushroom sticks 1 (such as black fungus mushroom sticks 1 in this case). However, in this case, the mushroom sticks 1 are first sterilized at 100℃ for 24 hours under normal pressure, and then the mushroom sticks 1 are removed from the sterilization furnace while still hot at a high temperature of 85℃. When transporting them to the mushroom shed 7 for stacking, the mushroom sticks 1 are still kept at a high temperature of 80℃. During transportation, they need to be covered with isolation cloth to prevent contamination.

[0018] To further achieve sterilization, after initial disinfection, fumigation disinfection is carried out using S10. Specifically, a first sealing film 4 is laid on the ground inside the mushroom shed 7. Several rows of first mushroom sticks 2 are stacked on the first sealing film 4 in a linear arrangement. Based on these first mushroom stick layers 2, several rows of mushroom stick piles 3 are formed, each consisting of several layers of stacked mushroom sticks 1, each layer composed of multiple mushroom sticks 1 arranged in a linear arrangement. The row spacing A between adjacent rows of mushroom stick piles 3 is 1 to 1.5 times the length L of the mushroom stick 1. The height of each mushroom stick pile 3 includes 9 to 12 layers of mushroom sticks 1. A second sealing film 5 is laid on top of the mushroom pile 3, and the periphery of the first sealing film 4 and the second sealing film 5 are sealed to form a closed mushroom pile. Preferably, the first sealing film 4 and the second sealing film 5 can be made of plastic sheeting, which is rolled up and sealed with clips to isolate air. This method utilizes the airtightness and isolation properties of the plastic sheeting to create a relatively closed and sterile environment for the mushroom pile 3. The first sealing film 4 and the second sealing film 5 significantly reduce the entry of airborne bacteria into the mushroom pile by preventing direct airflow, effectively reducing the risk of contamination of the mushroom sticks 1. After the pile is built, the temperature of the mushroom sticks 1 can be reduced from 80°C to 62°C for 15 hours, which can achieve sterilization conditions and achieve the sterilization purpose. Further, on the basis of the above, the piled mushroom sticks 3 is fumigated for disinfection. There are two methods: the first is as follows Figure 4The first method involves placing an aerosol disinfectant in a container 6 between two adjacent rows of mushroom log piles 3 for fumigation disinfection. This method primarily utilizes the aerosol disinfectant within the container 6, which disperses to sterilize the enclosed mushroom pile, providing a long-lasting disinfection effect. The second method involves pre-installing air slits at the front and rear ends of the enclosed mushroom pile, allowing the aerosol disinfectant to be actively introduced (by an air pump or blower) into the enclosed pile for fumigation disinfection. Preferably, the dosage of the aerosol disinfectant is 1-4 times per 10,000 mushroom logs 1. kg; The components of the aerosol disinfectant have strong oxidizing properties. These components can react with the cell membranes or cell walls of bacteria, viruses and other microorganisms in the air, destroying their structure and causing the microorganisms to lose their activity and die. Both of the above methods can allow the aerosol disinfectant to diffuse and cover the entire space area (i.e., the area enclosed by the mushroom stick pile 3 after the first sealing film 4 and the second sealing film 5 are sealed; the first method is slower, and the second is relatively faster), thereby achieving comprehensive disinfection of the air and object surfaces. It can also directly cover and act on the target area, such as the perforation of the mushroom bag, forming a mucous membrane, thereby achieving rapid and effective disinfection and sterilization, reducing the probability of the mushroom stick 1 being directly exposed to the air and causing contamination by miscellaneous bacteria. Due to the protection of the mucous membrane formed by the aerosol disinfectant on the surface of the mushroom stick 1, the risk of direct exposure to the air during the handling of the mushroom stick 1 is also reduced, thus reducing contamination by miscellaneous bacteria.

[0019] To verify the anti-contamination effect of the above sterilization and disinfection steps, the following experiment was conducted: (1) Experimental formula TSA medium formula: 5g tryptone, 2.5g yeast extract, 1g glucose, 15g agar, pH 7.0±0.2 (per liter); Formula for mushroom substrate 1: 93.1% hardwood sawdust, 6% wheat bran, and 0.9% light calcium carbonate. Mushroom bag dimensions: 14*55cm; Mushroom substrate 1: 42cm long; Weight: 1.4-1.5kg. Mushroom sticks 1 are bagged and sterilized according to standard procedures to produce mushroom stick 1.

[0020] (2) High-temperature pile building of mushroom sticks 1 Following the steps above, place the black fungus spawn 1 into a sterilizer for high-temperature sterilization. After sterilization, wait for the temperature inside the sterilizer to drop to 85℃ and remove the spawn while still hot. Transport the spawn to the mushroom shed 7 for stacking. During transportation, cover the spawn with plastic sheeting (to isolate it from the air). Before stacking the spawn 1, lay a layer of plastic film (i.e., the first sealing film 4) on the ground to prevent ground bacteria from contaminating the spawn 1. When stacking, arrange the sterilized spawn 1 in a straight line with a certain row spacing and stack height. Each stack contains 12,500-3,000 spawn spawn spawn spawn 3, and the volume of the stack is 5.04 m³. After stacking, cover the top of the stack with another layer of plastic film (i.e., the second sealing film 5). Fold the top and bottom plastic films together and clamp them tightly to seal them. When sealing the stack, leave a gap of about 30 cm at the front and back ends. Place the aerosol disinfectant in a container 6 between the stacks of spawn 3 for sterilization. Correspondingly, natural cooling eliminates the above-mentioned steps after pile building, and the mushroom sticks 1 are directly exposed to the air for natural cooling; (3) Different treatments were set up after the mushroom logs were piled up. This experiment set up five treatment methods after the mycelium log 1 was established: Treatment 1: High-temperature isolation, stacked in a straight line, use 4 kg of aerosol disinfectant per 10,000 mushroom sticks; Treatment 2: High-temperature isolation, stacked in a straight line, use 2 kg of aerosol disinfectant per 10,000 mushroom sticks; Treatment 3: High-temperature isolation, stacked in a straight line, use 1 kg of aerosol disinfectant per 10,000 mushroom sticks; Process 4 (CK): Natural cooling, stacked in a straight line; Process 5: Natural cooling, stack in a grid pattern.

[0021] The experiment was conducted in 3 replicates, with 500 sticks per replicate, corresponding to treatments one through five above, requiring a total of 7500 sticks; the number of colonies in different treatment spaces, the contamination status of stick 1 before inoculation, and the contamination status of stick 1 when removing the bag were recorded.

[0022] (4) Cleanliness test of the microbial stack space On days 8, 10, and 12 after the mycelium pile was established, two TSA culture media (culture medium ① and culture medium ②) were placed in the mycelium piles of treatments 1, 2, and 3 and in the outside air, respectively, and left to stand for 3 hours. The TSA culture media with attached bacteria were cultured in a constant temperature incubator at 28±1℃ and 75% humidity for 3 days (72 hours), and the number of colonies in the TSA culture media was observed.

[0023] (5) Observe the contamination status of mushroom stick 1 On days 5, 8, and 10 after the establishment of the mushroom substrate pile, the contamination status of mushroom substrate 1 under treatments 1, 2, 3, 4, and 5 was observed and recorded. When the mycelium of mushroom substrate 1 filled the bag and it was necessary to remove the bag and puncture the hole, the contamination status of mushroom substrate 1 was observed and recorded.

[0024] (6) Experimental results 6.1) Results of cleanliness test of the microbial contamination space After culturing TSA in a constant temperature incubator at 28±1℃ and 75% humidity for 3 days, the number of colonies was observed and recorded, as shown in the table below (contamination of culture medium under different amounts of aerosol disinfectant treatment):

[0025] 6.2) Observation of contamination status of mushroom stick 1 No contamination was found in the three treatments: high-temperature isolation fumigation, high-temperature isolation of the mushroom piles, stacking them in a straight line, and using aerosol disinfectant (corresponding to treatments one, two, and three). The mushroom sticks were naturally cooled and placed in a straight line. No aerosol disinfectant was used for fumigation of the mushroom sticks 1. On the 5th day after the mushroom sticks 1 were placed in a straight line, no contamination was found when the pile was turned over. On the 8th day, the pile was turned over again and some mushroom sticks 1 were found to be contaminated. The contamination was mainly on the surface of the mushroom bags, and the contamination diameter was 1-4cm. No contamination was found at the tied end. A total of 373 mushroom sticks 1 were investigated in the experiment, of which 25 mushroom sticks 1 were contaminated. The mushroom sticks were arranged in a grid pattern with natural cooling. No aerosol disinfectant was used to fumigate the sticks. After 10 days of arrangement, it was found that most of the mushroom sticks were contaminated, mainly at the needle holes of the mushroom bags on the surface of the sticks. The contamination at the tied ends was less. The contamination of the mushroom sticks was visible to the naked eye. The largest sticks were 8 cm in diameter and the smallest were 1 cm. A total of 162 mushroom sticks were investigated in the experiment. Only 12 sticks were uncontaminated at the time of inoculation. (7) Experimental conclusions Compared to air, the cleanliness of the black fungus spawn pile space was significantly improved after high-temperature isolation fumigation. The sterilization effect of different doses of aerosol disinfectant was: 4 kg > 2 kg > 1 kg. The sterilization effect of 4 kg and 2 kg aerosol disinfectant was not significantly different. After the spawn piles were exposed to the air and cooled naturally, and arranged in a straight line and a grid pattern for 10 days, the contamination rate of the spawn piles arranged in a straight line was 6.7%, while the contamination rate of the spawn piles arranged in a grid pattern was 92.6%. The contamination rate of the spawn piles arranged in a straight line was significantly lower than that of the spawn piles arranged in a grid pattern. Therefore, high-temperature isolation fumigation significantly reduced the number of miscellaneous bacteria in the air of the spawn pile and significantly reduced the contamination rate of the spawn piles.

[0026] After the pasteurization and hot handling in step S00, and the stacking and fumigation sterilization of the mushroom sticks 1 in step S10, the mushroom sticks 1 can be inoculated in step S20: the fumigated mushroom sticks 1 are transferred to the inoculation box under sealed conditions; the step of transferring the mushroom sticks 1 to the inoculation box is as follows: the transfer is carried out through a transfer chamber equipped with a disinfection function; the outer door of the transfer chamber is opened, the mushroom sticks 1 are placed in, and then closed, followed by aerosol and / or ultraviolet disinfection inside the chamber. After disinfection, the inner door is opened and the mushroom sticks 1 are sent into the inoculation box; the sealed conditions are usually achieved using a device equipped with The inoculation box uses a self-cleaning transfer container 6 for high-efficiency particulate air (HEPA) filters, or can be transferred as a whole using a sterile bag / box. Preferably, the interior of the inoculation box is a sterilized environment, which can be further configured as a sterile positive pressure environment. This positive pressure airflow prevents the infiltration of humid and hot air, inhibits mold growth, and effectively keeps dust, bacteria, odors, moisture, and contaminants outside the inoculation box. In addition, the inoculation box is equipped with at least one operating hole with an operating sleeve, an automatic punching device, and a microbial inoculation device; specifically, the microbial inoculation device is integrated into the automatic punching device. Inside, after punching, the inoculum is injected into the bottom of the hole; that is, the punching and inoculation operations are performed simultaneously. For example, this can be achieved using a punching and inoculation composite needle, which includes a hollow needle tube and a retractable spike. During operation, the spike extends first to pierce the bag and form an inoculation hole. After the spike reaches a preset depth or during retraction, a quantitative amount of inoculum is injected into the hole through the needle tube. Then, the entire composite needle is withdrawn from the substrate 1. Alternatively, other manual operation methods can be used through the operating hole, which will not be elaborated here. The punching device also includes a visual positioning sensor for identifying the substrate 1. The position is set to assist the automatic punching device in punching; the box is also equipped with a humidity sensor, whose data is connected to an external controller for display and alarm, so as to monitor the status inside the inoculation box. Furthermore, to achieve optimal matching of the entire process from the size of the mushroom stick 1 to the inoculation parameters to the punching execution, the automatic punching device includes a punching head driven by a servo motor (or cylinder, air pump). The punching depth, hole diameter, and hole spacing are automatically adjusted according to the length and diameter of the mushroom stick 1 based on the automatic calculation module of the external controller; wherein, in the automatic calculation module, the punching depth H satisfies:

[0027] in, The diameter of the mushroom log is 1. To optimize the constant, the range is 1~2 cm; When inoculating directly into the radial center of the substrate 1, the spawn can be directly inoculated into the core area of ​​the substrate 1. This area has the most stable substrate compaction, humidity, and temperature, and is least affected by external environmental fluctuations. This is conducive to the mycelium radiating outwards along the shortest path and in the most uniform way, quickly occupying nutrient space and forming a growth space advantage over other miscellaneous fungi. It is necessary to consider that there may be errors due to the deviation of the punching head during mechanical punching. At the same time, in order to better facilitate the initial germination of the spawn block and facilitate its initial respiration, the Δ value makes the spawn block inoculated in the hole slightly suspended. That is, the small space reserved between the bottom of the hole and the core of the substrate 1 introduces a small amount of air during inoculation, which can form a local microenvironment conducive to the colonization and germination of the spawn block.

[0028] Preferably, the aperture ranges from 3.6 to 4 cm; After determining the drilling depth H on the substrate 1, another parameter is the hole spacing S. Firstly, the ends of substrate 1 are usually the areas most prone to dehydration, compression, or contamination. To protect the structural integrity of substrate 1, drilling should be avoided in these areas. Furthermore, contaminants can easily invade through the gaps at the bag opening of substrate 1; inoculating away from the ends reduces the risk of direct contamination. Secondly, the mycelia germinating from each point after drilling should almost simultaneously and at a uniform rate cover the entire substrate 1, without significant growth retardation zones, thus allowing the mycelia to occupy the entire substrate in the shortest possible time. This requires the hole spacing S to meet the following conditions:

[0029] Where L is the length of the mushroom stick 1; E is the end distance, which is the distance between the holes at the outermost two ends and the two ends of the mushroom stick 1, which is 3~5 cm; N is the number of holes; Through the above methods, the driving device (servo motor or cylinder, air pump) in the automatic punching device drives the punching head in combination with preset algorithms and parameter adjustments, ensuring that every hole on each mushroom stick 1 has a consistent depth and precise spacing. At the same time, it can also be adjusted manually through the operation hole to completely solve the problem of inconsistent punching depth and random distribution, so that the growth process of the entire batch of mushroom sticks 1 is highly synchronized. At the same time, the precise hole depth and hole position data can be linked to control the subsequent inoculation amount (the amount of inoculum injected into each hole can be more precise) and sealing operation to improve the quality of mycelial cultivation.

[0030] S30: Through the operating hole, the automatic punching device punches holes in the inoculation box using the operating device, and then inoculates the spawn into the holes using the spawn inoculation device. The inoculation holes are then sealed by covering them with a spawn bag (i.e., covering with an outer tie, and sealing the spawn bag after covering with the outer tie; this solution is simple and efficient for black fungus spawn). Preferably, the inoculation box has two operating holes arranged opposite to each other and corresponding operating covers, forming a two-person collaborative workstation. The top of the box is equipped with a transparent observation window and a light, the bottom of the box is equipped with casters, and the side of the box is equipped with a sealed conveyor pipe interface for outputting the inoculated spawn bag 1. It also includes a sealed conveyor belt or trolley that cooperates with the conveyor pipe interface to transport the inoculated spawn bag 1 in a sealed manner to the culturing greenhouse for cultivation, i.e., step S40: the inoculated and sealed spawn bag 1 is removed from the inoculation box under sealed conditions and transported to the culturing greenhouse.

[0031] Example 2: The standardized execution procedure for punching holes in Example 1 is measurement-setting-execution. This involves manually or by sampling to measure the specifications of the mushroom sticks 1, pre-setting the average parameters into the system, and then executing the procedure using various execution devices. The advantages of this setup are low overall system cost, high reliability, simple debugging and maintenance, and low operator requirements. However, its disadvantage is rigid production; manual parameter adjustments and auxiliary operations may be needed to handle different situations. Therefore, to improve production flexibility and automatically adapt to different specifications and even different varieties of mushroom sticks 1, a sensing control unit is also installed in the inoculation box. After the mushroom sticks 1 are placed in the inoculation box, the sensing control unit automatically identifies the specifications of the mushroom sticks 1. The system automatically calculates and outputs the optimal number of inoculation holes, spatial layout, and depth information based on the specified parameters and the preset mycelial growth model in the sensing control unit. It then drives the automatic punching device and the mycelial inoculation device connected to the control unit to perform synchronous punching and inoculation operations on the mycelial sticks 1 according to the output information. This transforms the standardized execution procedure into a perception-decision-execution process, real-time sensing of the non-standard characteristics (such as diameter, length, and even density) of each mycelial stick 1, calling the mycelial growth model for calculation. This allows for handling complex material conditions and drives execution to achieve a more consistent mycelial growth rate. Therefore, the process of automatically calculating the optimal inoculation parameters based on the specified parameters and the preset mycelial growth model includes: S21: Obtain the effective length and diameter D of mushroom stick 1; S22: Based on the radial average growth rate of the mycelium of the target strain in the culture medium. Compared with the average longitudinal growth rate , and the preset time T for complete occupation of the target hyphae; S23: Calculate the number of inoculation points required for uniform distribution across the cross-section. Among them, the theoretical distance from any point on the cross-section to the nearest inoculation point is less than ,in, ; S24: Calculate the required number of inoculation rows in the longitudinal direction. Where the row spacing A satisfies: ; S25: Outputs the total number of inoculation wells N. .

[0032] In this way, it is ensured that after inoculation, within a preset time (i.e., occupation time T), the mycelium can grow evenly and synchronously from each inoculation point to the interior of the substrate 1. The purpose of step S23 is to ensure that when the mycelium grows from the evenly distributed points on the circumference towards the center of the substrate 1, the leading edge of the mycelium can converge at the center point within time T, thus ensuring that the mycelium can cover any blank area on the radius within a specified time. The purpose of step S24 is to ensure that when the mycelium grows from each row of inoculation points in the longitudinal distribution towards both ends, the leading edge of the mycelium can converge with the mycelium of the adjacent row within time T. The mycelium grows from each inoculation point in both forward and backward directions at the same time. Thus, steps S23 and S24 are used to pursue the highest utilization efficiency of the substrate 1.

[0033] This invention has been described through preferred embodiments. Those skilled in the art will understand that various changes or equivalent substitutions can be made to these features and embodiments without departing from the spirit and scope of the invention. This invention is not limited to the specific embodiments disclosed herein; other embodiments falling within the scope of the claims are also within the protection scope of this invention.

Claims

1. A method for preventing contamination during inoculation of edible mushroom substrate, characterized in that, Includes the following steps: S00: The mushroom sticks are first sterilized at 100℃ for 24 hours under normal pressure for preliminary disinfection. The sterilized mushroom sticks are then removed from the sterilization furnace while still hot at a temperature not lower than 85℃ and transported to the mushroom shed under covered conditions. S10: A first sealing film is laid on the ground inside the mushroom shed. Several rows of first mushroom sticks are stacked on the first sealing film in a linear arrangement. Based on the first mushroom sticks, several rows of mushroom stick piles are stacked up in sequence. A second sealing film is laid on top of the several rows of mushroom stick piles. The periphery of the first sealing film and the second sealing film is sealed to form a closed mushroom pile. An aerosol disinfectant is placed in a container and placed between two adjacent rows of mushroom stick piles for fumigation disinfection. S20: The fumigated and disinfected mushroom sticks are transferred to the inoculation box under sealed conditions; the inoculation box is a sterile environment after sterilization treatment, and the inoculation box is equipped with at least one operating hole with an operating sleeve, an automatic punching device, and a mushroom inoculation device; S30: Through the operating hole, the automatic punching device punches holes in the inoculation box, then the inoculation device inoculates the inoculation into the holes, and the inoculation holes are sealed by covering them with a mushroom bag. S40: After inoculation and sealing, remove the inoculation box under sealed conditions and transfer it to the mushroom cultivation shed.

2. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 1, characterized in that, In step S10, the row spacing A between two adjacent rows of mushroom stick piles is 1 to 1.5 times the length L of the mushroom sticks, and the height of the mushroom stick pile includes 9 to 12 layers of mushroom sticks.

3. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 2, characterized in that, In step S10, the amount of aerosol disinfectant used is 1 to 4 kg per 10,000 mushroom sticks.

4. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 3, characterized in that, In step S20, the step of transferring the mushroom sticks to the inoculation box is as follows: the transfer is carried out through a transfer chamber equipped with a disinfection function; the outer door of the transfer chamber is opened, the mushroom sticks are placed in, and then closed; subsequently, aerosol and / or ultraviolet disinfection is performed inside the chamber; after disinfection, the inner door is opened and the mushroom sticks are sent into the inoculation box.

5. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 4, characterized in that, In step S20, the automatic punching device includes a punching head driven by a servo motor, and the punching depth, hole diameter, and hole spacing are automatically adjusted according to the length and diameter of the mushroom stick; wherein, the punching depth H satisfies: in, The diameter of the mushroom log; To optimize the constant, the range is 1~2 cm; The aperture ranges from 3.6 to 4 cm; The hole spacing S satisfies: Where L is the length of the mushroom log; E is the end distance, which is the distance between the holes at the outermost two ends and the two ends of the mushroom log, which is 3~5 cm; and N is the number of holes.

6. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 5, characterized in that, In step S20, the inoculation device is integrated inside the automatic punching device, and the inoculum is injected into the bottom of the hole after punching is completed.

7. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 6, characterized in that, In step S20, the punching device also includes a visual positioning sensor for identifying the position of the mushroom sticks; the box is also equipped with a humidity sensor, whose data is connected to an external controller for display and alarm.

8. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 7, characterized in that, In step S30, the inoculation box is provided with two oppositely arranged operating holes and corresponding operating sleeves, forming a two-person collaborative workstation; the top of the box is provided with a transparent observation window and a lighting lamp, the bottom of the box is provided with casters, and the side of the box is provided with a closed conveyor pipe interface for outputting the inoculated mushroom sticks. It also includes a closed conveyor belt or mushroom cultivation cart that cooperates with the conveyor pipe interface, for transferring the inoculated mushroom sticks in a closed manner to the mushroom cultivation shed.

9. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 1, characterized in that, In step S20, the inoculation box is also equipped with a sensing control unit. After the mushroom stick is placed in the inoculation box, the sensing control unit automatically identifies the specifications of the mushroom stick and, based on the specifications and the mycelial growth model preset in the sensing control unit, automatically calculates and outputs the optimal number of inoculation holes, spatial layout, and depth information; drives the automatic punching device and the mycelial inoculation device connected to the control unit to perform synchronous punching and inoculation operations on the mushroom stick according to the output information.

10. The method for preventing contamination during inoculation of edible mushroom substrate according to claim 9, characterized in that, The process of automatically calculating the optimal inoculation parameters based on specification parameters and a preset mycelial growth model includes: S21: Obtain the effective length (L) and diameter (D) of the mushroom log; S22: Based on the radial average growth rate of the mycelium of the target strain in the culture medium. Compared with the average longitudinal growth rate , and the preset time T for complete occupation of the target hyphae; S23: Calculate the number of inoculation points required for uniform distribution across the cross-section. Among them, the theoretical distance from any point on the cross-section to the nearest inoculation point is less than ,in, ; S24: Calculate the required number of inoculation rows in the longitudinal direction. Where the row spacing A satisfies: ; S25: Outputs the total number of inoculation wells N. .

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