Method for on-line correction of inoculation ratio of multi-strains for special equipment for fermented food production

Abstract

The present application relates to a multi-strain inoculation ratio online correction method for special equipment for fermented food production, and belongs to the technical field of fermented food production equipment and inoculation control in fermentation process, comprising: the equipment comprises a main fermentation cavity, a bypass test supplement cavity, a sterile sampling channel, at least two sterile inoculum channels respectively communicating different strain storage liquid units, and a waste liquid sterilization channel; when the inoculation ratio of any target strain in the main fermentation cavity is lower than the set lower limit of the corresponding fermentation stage, a preset sampling amount of fermentation material is sent into the bypass test supplement cavity through the sterile sampling channel, and the target strain liquid is added to the bypass test supplement cavity for test supplement reaction; the release judgment result is formed according to the pH change rate, the gas production rate, the oxidation-reduction potential change rate and the target metabolite concentration change rate, and the corresponding sterile inoculum channel is opened or closed accordingly. The method can realize online correction of the low limit deviation of the inoculation ratio of the target strain, improve the stability, controllability and product batch consistency of the fermentation process.

Description

Technical Field

[0001] This invention relates to the field of fermented food production equipment and fermentation process inoculation control technology, specifically to a method for online correction of multi-strain inoculation ratio for specialized fermented food production equipment. Background Technology

[0002] In the industrial production of fermented foods, existing technologies typically involve centralized control of raw materials and microbial strains through fermentation tanks, inoculation pipelines, and temperature and pH monitoring units. The focus is on ensuring a stable fermentation environment and achieving predetermined batch process parameters. For example, Chinese patent publication CN115851437A relates to fermentation equipment or control structures related to fermented food production, which can improve the continuity of the fermentation process by addressing aspects such as temperature, stirring, aeration, and sampling. Chinese patent publication CN116445296A relates to the regulation of fermentation conditions using multiple or mixed microbial strains, which can improve the quality of fermented products through strain combinations, culture conditions, or process monitoring. These technologies indicate that in recent years, fermented food production has gradually shifted from experience-based inoculation to equipment-based, parameterized, and automated control, and has begun to focus on the impact of microbial community composition on acidity, gas generation, and flavor metabolism.

[0003] However, in fermentation systems involving multiple microbial species, different species exhibit variations in growth rates, substrate utilization pathways, acid and gas production capabilities, and metabolite generation directions. Simply relying on initial inoculum levels or overall fermenter parameters such as pH, temperature, pressure, and dissolved oxygen is insufficient to accurately reflect the low actual proportion of a particular target species at a specific fermentation stage. When localized nutrient changes, decreased microbial activity, increased interspecies competition, or sampling delays occur during production, directly introducing the target species into the main fermentation system may lead to incompatibility with the existing microbial community or further deviate from the original inoculum ratio, resulting in rapid acidification, abnormal gas production, deviations in flavor compound generation, and increased risk of contamination. Especially in continuous or semi-continuous fermentation systems, the materials within the main fermentation chamber are in a dynamic state, making it difficult to directly determine the inoculum quantity, timing, and safety using static process tables.

[0004] Furthermore, existing fermentation control schemes mostly focus on closed-loop regulation of overall environmental parameters within the main fermentation chamber, lacking a mechanism for small-scale, isolated, and waste-discharge-enabled trial addition verification of the same batch of fermented materials before actual addition of bacteria. They also lack technical means to comprehensively determine release based on pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate. Therefore, how to perform online correction for target bacteria below a set lower limit without compromising the stability of the main fermentation system, and how to eliminate the risk of unsuitable or excessive addition of bacteria before addition, have become technical problems that specialized equipment for fermented food production needs to solve. Summary of the Invention

[0005] The purpose of this invention is to provide an online correction method for the inoculation ratio of multiple microorganisms in specialized equipment for fermented food production, thereby addressing some of the drawbacks and shortcomings pointed out in the background art.

[0006] The present invention adopts the following technical solution to solve the above-mentioned technical problems:

[0007] The equipment includes a main fermentation chamber, a bypass test chamber connected to the main fermentation chamber, a sterile sampling channel, at least two sterile replenishment channels respectively connected to different strains of liquid storage units, and a waste liquid sterilization channel.

[0008] When the inoculation ratio of any target strain in the main fermentation chamber is lower than the set lower limit of the corresponding fermentation stage, the fermentation material of the preset sampling amount is sent into the bypass test chamber through the aseptic sampling channel, and the bacterial solution of the target strain is added to the bypass test chamber for test reaction.

[0009] Release determination results are formed based on the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate in the bypass test chamber. When the release determination results are within the preset release range, the corresponding aseptic replenishment channel is opened to replenish the bacterial solution of the target strain into the main fermentation chamber. When the release determination results are not within the preset release range, the corresponding aseptic replenishment channel is closed, and the material in the bypass test chamber is discharged through the waste liquid sterilization channel.

[0010] Furthermore, the bypass test replenishment chamber is divided into a baseline zone, a test replenishment zone, and an excess zone. Fermentation materials from the same batch, drawn through the aseptic sampling channel, enter the baseline zone, the test replenishment zone, and the excess zone, respectively. No bacterial solution of the target strain is added to the baseline zone. A first preset inoculation amount of bacterial solution of the target strain is added to the test replenishment zone. A second preset inoculation amount of bacterial solution of the target strain is added to the excess zone. The second preset inoculation amount is greater than the first preset inoculation amount. When the release determination result of the test replenishment zone is within the preset release range, and the excess zone does not show a pH change rate exceeding a preset mutation threshold, a gas production rate exceeding a preset gas production upper limit, or a target metabolite concentration change direction opposite to the preset target direction relative to the baseline zone, the corresponding aseptic replenishment channel is opened.

[0011] Furthermore, the bypass test chamber is equipped with a cell retention component, which allows soluble substrates and metabolites to pass through while blocking the target strain from entering the aseptic replenishment channel from the bypass test chamber. The bacterial solution of the target strain first contacts the fermentation material in the cell retention component. When the release determination result is within the preset release range, the corresponding aseptic replenishment channel is opened. When the release determination result is not within the preset release range, the material in the cell retention component is discharged into the waste liquid sterilization channel.

[0012] Furthermore, when the inoculation ratio of at least two target strains is lower than the set lower limit of the corresponding fermentation stage, the aseptic replenishment channel is opened one by one according to the bypass replenishment judgment result; wherein, before the bacterial solution of one target strain is replenished into the main fermentation chamber and verified by the bypass replenishment chamber, the aseptic replenishment channel corresponding to other target strains remains closed, so as to avoid the secondary deviation of the inoculation ratio caused by the simultaneous replenishment of multiple strains.

[0013] Furthermore, the bacterial cell retention component has a retention state and a release state; in the retention state, the target bacterial cells are confined within the bacterial cell retention component, and the exchange of soluble substrates and metabolites within the bypass test chamber is allowed; when the release determination result is within the preset release range, the bacterial cell retention component switches to the release state, allowing the retained target bacterial cells to enter the aseptic replenishment channel along with the corresponding target bacterial solution.

[0014] Furthermore, the microbial retention component is connected to a backwash branch; when the release determination result is not within the preset release range, the corresponding aseptic replenishment channel is first closed, and then the target microbial strain and residual fermentation material in the microbial retention component are pushed to the waste liquid sterilization channel through the backwash branch.

[0015] Furthermore, the bacterial retention component is divided into a proximal retention zone and a distal retention zone in the same replenishment reaction. The proximal retention zone is located upstream of the direction in which the target bacterial culture enters, and the distal retention zone is located downstream of the direction in which the target bacterial culture enters. The target bacterial culture first enters the proximal retention zone. When the release determination result is within the preset release range, the distal retention zone is released first, and the proximal retention zone is released after a preset delay time, so that the target bacterial culture entering the aseptic replenishment channel is replenished in segments.

[0016] Furthermore, when the aseptic replenishment channels are opened one by one, the aseptic replenishment channel corresponding to the target strain whose time from the addition of the target strain solution to the first time meeting the preset release range is opened first. After the target strain solution is replenished into the main fermentation chamber, if the verification result of the bypass test replenishment channel is still within the preset release range, the aseptic replenishment channel corresponding to the next target strain is opened.

[0017] Furthermore, during the process of opening the aseptic replenishment channels one by one, if the verification result of the bypass test replenishment channel after the bacterial solution of the current target strain is replenished into the main fermentation chamber is not within the preset release range, the aseptic replenishment channel corresponding to the subsequent target strain remains closed, and the aseptic replenishment channel corresponding to the current target strain is switched to an intermittent replenishment state executed according to a preset single replenishment amount and a preset replenishment interval, or switched to a stopped replenishment state.

[0018] Furthermore, before opening the aseptic replenishment channel corresponding to the next target strain, the main fermentation chamber material sample containing the target strain and the bacterial solution of the next target strain are added together to the bypass test replenishment chamber for sequential compatibility verification; when the verification result of the bypass test replenishment chamber is still within the preset release range, the aseptic replenishment channel corresponding to the next target strain is opened; when the verification result of the bypass test replenishment chamber is not within the preset release range, the aseptic replenishment channel corresponding to the next target strain remains closed.

[0019] This invention establishes a bypass test chamber outside the main fermentation chamber and utilizes a sterile sampling channel to introduce the same batch of fermentation material into the bypass test chamber for target microbial supplementation. This allows the supplementation determination to be completed in an isolated, small-scale environment. The target microbial strain is only introduced into the main fermentation chamber through the corresponding sterile supplementation channel when the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate within the bypass test chamber all meet the preset release range. This avoids imbalances in the main system caused by directly supplementing microorganisms based on a single detection parameter or manual experience.

[0020] When the release determination result is not within the preset release range, this invention closes the corresponding aseptic replenishment channel and discharges the material in the bypass test replenishment chamber through the waste liquid sterilization channel. This isolates and eliminates unsuitable bacterial solutions and fermentation materials before replenishment, reducing the risk of contamination, metabolic mutation, and over-replenishment. For equipment structures with at least two bacterial storage units and corresponding aseptic replenishment channels, this method can achieve online correction of the lower limit deviation of the target bacterial inoculation ratio, improving the stability, controllability, and batch consistency of multi-species fermentation processes. Attached Figure Description

[0021] Figure 1This is a flowchart of the online correction method for the inoculation ratio of multiple bacterial strains according to the present invention.

[0022] Figure 2 This is a diagram showing the connection between the bypass test area and the channel in Embodiment 1 of the present invention.

[0023] Figure 3 This is a diagram showing the evaluation and threshold determination of the trial release area in Embodiment 1 of the present invention.

[0024] Figure 4 This is a diagram showing the segmented release volume of the bacterial cell retention component in Embodiment 1 of the present invention.

[0025] Figure 5 This is a comparison chart of the lower limits of inoculation ratios for multiple target bacterial strains in Example 2 of the present invention.

[0026] Figure 6 This is a diagram showing the order of adding multiple target bacterial strains in Example 2 of the present invention.

[0027] Figure 7 This is a flowchart of intermittent supplementation and sequential compatibility verification in Embodiment 2 of the present invention. Detailed Implementation

[0028] As attached Figure 1 As shown, in one specific embodiment, the online correction method for the multi-strain inoculation ratio of specialized equipment for fermented food production can be implemented using a main fermentation chamber, a bypass test chamber, a sterile sampling channel, a sterile inoculation channel, and a waste liquid sterilization channel. The main fermentation chamber contains the fermentation materials during the fermentation process. The bypass test chamber is connected to the main fermentation chamber and is used for small-scale test inoculation verification without directly interfering with the fermentation state of the main fermentation chamber. The sterile sampling channel is used to introduce the fermentation materials from the main fermentation chamber into the bypass test chamber. At least two sterile inoculation channels are provided, each connected to a different strain storage unit, to allow for independent inoculation control for different target strains. Sterile valves, transfer pumps, and online sterilization interfaces can be installed on the sterile sampling channel and the sterile inoculation channel respectively to keep the channels closed during sampling and inoculation. The waste liquid sterilization channel is used to receive materials that fail the release judgment in the bypass test chamber and to sterilize the discharged materials. The waste liquid sterilization channel can use one or more of the following methods to treat the discharged materials: heat sterilization, ultraviolet sterilization, or chemical sterilization.

[0029] During fermentation, the inoculation ratio of each target strain in the main fermentation chamber can be detected in real time or at preset intervals. The inoculation ratio can be obtained through an online detection unit installed in the main fermentation chamber. This online detection unit may include an optical detection module, a fluorescence recognition module, a rapid nucleic acid detection module, or a microbial image recognition module. The system determines the inoculation ratio of each target strain at the current fermentation stage based on the detection signal and pre-stored strain identification information. When the inoculation ratio of any target strain falls below the set lower limit for the corresponding fermentation stage, the system sends a preset sample amount of fermentation material to the bypass test chamber through a sterile sampling channel. Simultaneously, the bacterial solution of the corresponding target strain is added to the bypass test chamber, allowing the bacterial solution of the target strain and the fermentation material to undergo a test reaction in the bypass environment. This method allows for pre-judgmentation of the impact of adding the target strain to the fermentation system before formally adding bacteria to the main fermentation chamber, reducing the risk of fermentation state fluctuations caused by direct bacterial addition.

[0030] During the trial replenishment reaction, the system collects the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate within the bypass trial replenishment chamber, and generates a release judgment result based on these parameters. The preset release ranges for these parameters can be pre-stored in the control unit according to the type of fermented food, fermentation stage, and target microbial strain. The system compares the collected pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate with their corresponding preset release ranges. When all parameters are within their respective preset release ranges, a release judgment result is generated indicating that the fermentation system is within the preset release range; when any parameter is outside the preset release range, a release judgment result is generated indicating that the fermentation system is outside the preset release range. When the release judgment result is within the preset release range, it indicates that the addition of the target microbial strain's culture can maintain the fermentation system in an acceptable state. The system then opens the corresponding aseptic replenishment channel and adds the target microbial strain's culture into the main fermentation chamber. When the release judgment result is not within the preset release range, it indicates that the trial replenishment may cause an abnormal fermentation state. The system closes the corresponding aseptic replenishment channel to prevent the target strain of bacteria from entering the main fermentation chamber, and discharges the material in the bypass trial replenishment chamber through the waste liquid sterilization channel.

[0031] In a further specific implementation, the bypass test chamber can be divided into a baseline zone, a test chamber, and an excess zone according to the test chamber function. Fermentation materials from the same batch, drawn through a sterile sampling channel, enter the baseline zone, test chamber, and excess zone respectively, ensuring consistency in the source of fermentation materials in the three zones and reducing the impact of batch differences on the judgment results. The baseline zone contains no target strain culture and reflects the basic changes in the fermentation material before the target strain is added. The test chamber contains a first preset inoculum amount of target strain culture and simulates the normal correction state after adding the target strain to the main fermentation chamber. The excess zone contains a second preset inoculum amount of target strain culture, set higher than the first preset inoculum amount, to observe whether an increase in the target strain addition will cause abnormal fluctuations in the fermentation system. A pH sensor, a gas generation detector, a redox potential detector, and a target metabolite detector can be installed in the baseline zone, test chamber, and excess zone respectively, to obtain test chamber reaction data for each zone under the same sampling source. When the release judgment result of the test replenishment zone is within the preset release range, and the excess zone does not show a pH change rate exceeding the preset mutation threshold, a gas production rate exceeding the preset gas production upper limit, or a target metabolite concentration change direction opposite to the preset target direction, the system opens the corresponding aseptic replenishment channel to replenish the target bacterial culture into the main fermentation chamber.

[0032] In another specific embodiment, a cell retention component is provided within the bypass replenishment chamber. This component forms a localized constraint area during the target bacterial culture replenishment process, allowing soluble substrates and metabolites to exchange between the retention component and the bypass replenishment chamber, while simultaneously preventing the target bacterial culture from entering the aseptic replenishment channel from the bypass chamber. The retention component can be a container structure with a microporous membrane, filter, or hollow fiber membrane. The microporous membrane, filter, or hollow fiber membrane restricts the passage of the target bacterial culture while allowing the passage of soluble substrates and metabolites. The target bacterial culture first enters the retention component and contacts the fermentation material within it, allowing for the acquisition of replenishment reaction data while restricting bacterial escape. When the release determination result is within a preset release range, the system opens the corresponding aseptic replenishment channel to perform subsequent replenishment operations. When the release determination result is not within the preset release range, the system will not allow the bacterial liquid of the target strain to enter the main fermentation chamber. Instead, it will discharge the material retained in the bacterial cell component into the waste liquid sterilization channel, thereby reducing the risk of bacterial cells and materials that have not passed the trial and replenishment verification affecting the main fermentation system.

[0033] In specific implementations requiring correction for multiple microbial strains, when the inoculation ratios of at least two target microbial strains are both below the set lower limit for the corresponding fermentation stage, the system does not simultaneously open multiple aseptic replenishment channels. Instead, it opens the corresponding aseptic replenishment channels one by one based on the bypass replenishment judgment results. After the bacterial solution of one target microbial strain is replenished into the main fermentation chamber, it needs to be verified through the bypass replenishment channel. During verification, the system obtains the replenished fermentation material from the main fermentation chamber and sends it to the bypass replenishment channel, forming a verification judgment result according to the same release judgment parameters and judgment rules as the trial replenishment reaction. Before completing the replenishment and verification of the target microbial strain, the aseptic replenishment channels corresponding to other target microbial strains remain closed. This control method can avoid mutual interference after multiple target microbial strains are replenished simultaneously, reducing the possibility of secondary deviations in the inoculation ratio within the main fermentation chamber.

[0034] In a further specific implementation, the bypass test chamber can be divided into a baseline zone, a test zone, and an excess zone. Fermentation materials from the same batch, drawn from the main fermentation chamber via a sterile sampling channel, enter the baseline zone, test zone, and excess zone respectively, ensuring that the fermentation materials in the three zones have the same source and reducing the impact of batch differences on the test zone determination. The baseline zone does not contain the target strain's bacterial solution and is used to obtain the basic changes in the fermentation material before the target strain is added. The test zone contains a first preset inoculum amount of the target strain's bacterial solution to simulate the reaction state after the target strain is added according to the corrected dosage. The excess zone contains a second preset inoculum amount of the target strain's bacterial solution, which is an increase relative to the first preset inoculum amount, and is used to verify whether increasing the target strain addition will cause adverse fluctuations in the fermentation system. The detection parameters from the baseline zone, test zone, and excess zone are all transmitted to the control unit, which generates the corresponding release determination results. When the release judgment result of the test replenishment zone is within the preset release range, and the excess zone does not show a pH change rate exceeding the preset mutation threshold, a gas production rate exceeding the preset gas production upper limit, or a target metabolite concentration change direction opposite to the preset target direction, the system opens the corresponding aseptic replenishment channel, allowing the target bacterial culture to be replenished into the main fermentation chamber.

[0035] In another specific embodiment, a cell retention component is provided within the bypass test replenishment chamber. This component creates a controlled contact area within the chamber, confining the target bacterial strain within it during the replenishment phase while allowing exchange of soluble substrates and metabolites between the component and the material within the chamber. The retention component can be a detachable retention box, membrane bag, or retention tube, with an isolation structure between it and the bypass test replenishment chamber facilitating the exchange of soluble substances. The target bacterial culture first enters the retention component and contacts the fermentation material within it, thus obtaining replenishment reaction data without the target bacterial strain directly entering the aseptic replenishment channel. When the release determination result falls within a preset release range, the system opens the corresponding aseptic replenishment channel to execute subsequent replenishment operations. When the release judgment result is not within the preset release range, the system does not allow the target strain of bacterial solution to be added to the main fermentation chamber. Instead, it discharges the material retained in the bacterial component into the waste liquid sterilization channel, and the waste liquid sterilization channel discharges the material that has not passed the trial replenishment verification.

[0036] In a specific implementation where simultaneous correction and judgment are required for multiple microbial strains, when the inoculation ratio of at least two target microbial strains is lower than the set lower limit for the corresponding fermentation stage, the system controls the opening of the corresponding aseptic inoculation channel one by one based on the bypass test inoculation judgment results for each target microbial strain. After the bacterial solution of one target microbial strain is added to the main fermentation chamber, the system verifies the fermentation system in the main fermentation chamber through the bypass test inoculation channel. The verification judgment result is formed in the same way as the release judgment result, both based on whether the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate are within the corresponding preset release range. Before the target microbial strain is added and the verification judgment result is obtained, the aseptic inoculation channels corresponding to other target microbial strains remain closed. Through the above-mentioned control method of adding and verifying one by one, the mutual influence caused by the simultaneous addition of multiple target microbial strains can be avoided, and the risk of secondary deviation of the inoculation ratio in the main fermentation chamber can be reduced.

[0037] In a further specific implementation, when multiple aseptic replenishment channels need to be opened one by one, the system determines the replenishment order based on the time required for each target strain to reach the preset release range in the bypass test replenishment chamber. Specifically, from the moment the bacterial solution of the target strain is added to the bypass test replenishment chamber, the system continuously acquires the release judgment parameters in the bypass test replenishment chamber and records the reaction time corresponding to the first time each target strain meets the preset release range. The system prioritizes opening the aseptic replenishment channel corresponding to the target strain with the shorter reaction time, allowing the bacterial solution of that target strain to be replenished into the main fermentation chamber first. When the reaction times corresponding to two target strains are the same or difficult to distinguish, the system opens the corresponding aseptic replenishment channel according to the pre-stored priority order of the target strains. After the bacterial solution of the target strain is replenished into the main fermentation chamber, the system again verifies the fermentation material in the main fermentation chamber through the bypass test replenishment chamber. When the verification judgment result is still within the preset release range, it indicates that the replenishment has not caused the fermentation system to deviate from the allowable state, and the system then opens the aseptic replenishment channel corresponding to the next target strain.

[0038] During the sequential opening of the aseptic replenishment channels, the system continuously assesses the fermentation status after the current target strain is added. If the verification result of the bypass test replenishment channel is not within the preset allowable range after the current target strain's culture is added to the main fermentation chamber, it indicates that the current addition has caused a deviation risk in the fermentation system. In this case, the system keeps the aseptic replenishment channel corresponding to subsequent target strains closed to prevent further fluctuations in the fermentation status caused by adding other target strains. Simultaneously, the system switches the aseptic replenishment channel corresponding to the current target strain to intermittent replenishment mode, making minor adjustments according to the preset single replenishment amount and preset replenishment interval, or switches the aseptic replenishment channel corresponding to the current target strain to stop replenishment mode to restore the strain ratio and fermentation parameters in the main fermentation chamber to the allowable range. In intermittent replenishment mode, the system verifies the result of each replenishment through the bypass test replenishment channel, and only allows the next replenishment to proceed if the verification result is within the preset allowable range.

[0039] Before opening the aseptic replenishment channel for the next target strain, the system can perform a sequential compatibility check. Specifically, a fermentation material sample containing the target strain has been added is taken from the main fermentation chamber, and this sample is added together with the bacterial solution of the next target strain to the bypass test replenishment chamber. This allows the bacterial solution of the next target strain to undergo a test replenishment reaction in a material environment that already contains the influence of the previous target strain. The system determines whether the next target strain is suitable for further addition based on the check result in the bypass test replenishment chamber. The sequential compatibility check uses the same judgment parameters and preset release range as the release judgment, and the check result serves as the basis for whether to open the aseptic replenishment channel for the next target strain. When the check result is still within the preset release range, the system opens the aseptic replenishment channel for the next target strain. When the check result is not within the preset release range, the system keeps the aseptic replenishment channel for the next target strain closed, thereby avoiding incompatible effects when different target strains are added sequentially.

[0040] Example 1:

[0041] This embodiment provides a method for online correction of multi-strain inoculation ratios in specialized equipment for fermented food production. This method can be executed by specialized equipment for fermented food production installed by a specific unit. The equipment includes a main fermentation chamber, a bypass test chamber, an aseptic sampling channel, at least two aseptic inoculation channels, a waste liquid sterilization channel, a strain storage unit, and a control unit. The main fermentation chamber contains the fermentation material. The bypass test chamber is connected to the main fermentation chamber and is used for small-scale test inoculation verification before the target strain is officially added to the main fermentation chamber. The aseptic sampling channel is used to introduce the fermentation material from the main fermentation chamber into the bypass test chamber. The aseptic inoculation channels are connected to different strain storage units. The waste liquid sterilization channel is used to receive bypass material that fails the release criteria and sterilize and discharge it.

[0042] In this embodiment, the fermentation material in the main fermentation chamber is 1200L, and the fermentation material is a mixed fermentation substrate used in the production process of a certain fermented food. The control unit detects the inoculation ratio of each target strain in the main fermentation chamber through an online detection unit. The online detection unit may include an optical detection module, a fluorescence recognition module, a rapid nucleic acid detection module, or a microbial image recognition module. When Lactobacillus plantarum is used as the target strain, the current inoculation ratio of Lactobacillus plantarum is detected to be 17.8%, which corresponds to a lower limit of 20.0% for the fermentation stage. The control unit determines that the inoculation ratio of Lactobacillus plantarum is lower than the lower limit and a bypass test is required for verification.

[0043] like Figure 2 As shown, a closed bypass test replenishment path is formed between the main fermentation chamber, the bypass test replenishment chamber, the Lactobacillus plantarum storage unit, the cell retention component, and the waste liquid sterilization channel. Figure 2By describing the sampling flow, inoculum replenishment flow, retention flow, and backwash flow, the process is illustrated: fermentation material first enters the bypass test replenishment chamber from the main fermentation chamber, then the target bacterial culture is supplied to the bypass test replenishment chamber by the *Lactobacillus plantarum* storage unit, and after passing the release judgment, it enters the corresponding aseptic inoculum replenishment channel through the bacterial retention component; when the release judgment fails... Figure 2 The reverse flushing path is used to guide the material retained in the bacterial cell component to the waste liquid sterilization channel, thereby preventing unvalidated bacterial cells from entering the main fermentation chamber.

[0044] The control unit activates the aseptic sampling channel, transferring 900 mL of fermentation material from the main fermentation chamber to the bypass test replenishment chamber. The bypass test replenishment chamber is divided into a baseline zone, a test replenishment zone, and an excess zone according to the test replenishment function. Each zone receives 300 mL of the same batch of fermentation material to minimize the impact of batch differences on the judgment results. No *Lactobacillus plantarum* culture is added to the baseline zone to obtain the basic changes in the fermentation material without *Lactobacillus plantarum* addition. 12 mL of *Lactobacillus plantarum* culture is added to the test replenishment zone as the first preset inoculum amount to simulate a normal correction state. 18 mL of *Lactobacillus plantarum* culture is added to the excess zone as the second preset inoculum amount to verify whether increasing the *Lactobacillus plantarum* addition causes adverse fluctuations in the fermentation system.

[0045] During the bypass replenishment reaction, pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate were collected in the baseline zone, replenishment zone, and excess zone, respectively. The control unit first converted the above four types of online detection parameters into dimensionless evaluation values ​​between 0 and 1, then set weights according to the degree of influence of each parameter on fermentation stability and performed weighted summation to obtain the release evaluation value for the replenishment zone. This release evaluation value is used to determine whether the addition of *Lactobacillus plantarum* culture will maintain the fermentation system in an acceptable state.

[0046] In this embodiment, the release evaluation value of the trial replenishment area is determined by the following formula:

[0047]

[0048] Wherein, B represents the release evaluation value of the test supplementation zone; Uph represents the pH change rate evaluation value; Ug represents the gas production rate evaluation value; Uo represents the redox potential change rate evaluation value; Um represents the target metabolite concentration change rate evaluation value; 0.35, 0.25, 0.20, and 0.20 represent the preset weights of pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate in the release evaluation, respectively. The above weighting settings ensure that pH change rate and gas production rate have a relatively high weight in the fermentation stability assessment, while also taking into account the indicative role of redox potential change rate and target metabolite concentration change rate in the fermentation state.

[0049] like Figure 3As shown, all four individual evaluation values ​​are above the release threshold of 0.80, and the overall evaluation B is also above the release threshold. Figure 3 By placing the evaluation of pH change rate, gas production rate, potential change rate, metabolite change rate, and comprehensive evaluation B within the same evaluation coordinate system, the release logic of the control unit is reflected by both the range of individual parameters and the threshold of comprehensive evaluation, thus avoiding the decision to release bacteria based solely on a single fermentation parameter.

[0050] In a single trial reaction, the control unit obtained the following evaluation values ​​for the trial reaction zone: pH change rate 0.88, gas production rate 0.82, redox potential change rate 0.90, and target metabolite concentration change rate 0.85. The preset release threshold was 0.80. Substituting these data into the calculation formula, we obtain:

[0051]

[0052] Since 0.863 is greater than 0.80, the control unit determines that the sample is within the preset release range. Simultaneously, the control unit compares the detection results of the excess zone with those of the baseline zone, confirming that the excess zone, relative to the baseline zone, does not exhibit a pH change rate exceeding the preset mutation threshold, a gas production rate exceeding the preset upper limit of gas production, or a target metabolite concentration change direction opposite to the preset target direction. Therefore, it allows the *Lactobacillus plantarum* culture to be added to the main fermentation chamber via the corresponding aseptic replenishment channel.

[0053] In this embodiment, a cell retention component is provided within the bypass test replenishment chamber. This cell retention component can be a retention box, membrane bag, or retention tube with a microporous isolation structure. The cell retention component allows the exchange of soluble substrates and metabolites between the bypass test replenishment chamber and the cell retention component, while preventing *Lactobacillus plantarum* cells from directly entering the aseptic replenishment channel during the test replenishment stage. The *Lactobacillus plantarum* bacterial solution first enters the cell retention component and comes into contact with the fermentation material within it, enabling the control unit to obtain test replenishment reaction data while restricting cell escape.

[0054] The cell retention component has a retention state and a release state. In the retention state, *Lactobacillus plantarum* cells are confined within the cell retention component, allowing for the exchange of soluble substrates and metabolites. When the release determination result is within the preset release range, the control unit switches the cell retention component from the retention state to the release state, allowing the retained *Lactobacillus plantarum* cells to enter the aseptic replenishment channel along with the corresponding *Lactobacillus plantarum* bacterial solution. If the release determination result is not within the preset release range, the control unit first closes the corresponding aseptic replenishment channel, and then pushes the *Lactobacillus plantarum* cells and residual fermentation material in the cell retention component to the waste liquid sterilization channel via a backwashing branch, where they are sterilized and discharged.

[0055] To reduce the risk of fluctuations in the fermentation state of the main fermentation chamber caused by a single release, the cell retention component is divided into a proximal retention zone and a distal retention zone in the same replenishment reaction. The proximal retention zone is located upstream of the Lactobacillus plantarum culture in the direction of entry, and the distal retention zone is located downstream. The Lactobacillus plantarum culture first enters the proximal retention zone and then the distal retention zone. When the release determination result is within the preset release range, the control unit first releases the distal retention zone, and the proximal retention zone is released after a preset delay time, so that the Lactobacillus plantarum culture entering the aseptic replenishment channel is replenished in stages.

[0056] like Figure 4 As shown, the distal retention zone is the first release zone, with a release time of 0 min and a release volume of 72 mL; the proximal retention zone is the delayed release zone, with a release time of 3 min and a release volume of 48 mL. Figure 4 The timing difference of the segmented replenishment is expressed by the correspondence between the release segment and the release volume, so that the total liquid holding capacity of the bacterial cell retention component does not enter the main fermentation chamber all at once, but is divided into two stages, namely, early release and delayed release, after the release judgment is passed, thereby weakening the instantaneous impact of bacterial replenishment.

[0057] In this embodiment, the total liquid holding capacity of the bacterial cell retention component is 120 mL. The volume distribution ratio of the distal retention zone in the bacterial cell retention component is 60%, and the volume distribution ratio of the proximal retention zone is 40%. The delayed release time of the proximal retention zone is 3 min. The volume of bacterial solution released first from the distal retention zone is determined by the following formula:

[0058]

[0059] Where Vf represents the volume of bacterial solution released first in the distal retention zone; Vh represents the total liquid holding capacity of the bacterial retention component; and Fd represents the volume distribution ratio of the distal retention zone within the bacterial retention component. The derivation logic of this formula is that the total amount of bacterial solution within the bacterial retention component is distributed in the proximal and distal retention zones according to a preset volume distribution ratio. The distal retention zone releases first, therefore the volume released first is determined by the total liquid holding capacity and the distal volume distribution ratio.

[0060] Substituting the total liquid holding capacity of the bacterial cell retention component (120 mL) and the volume distribution ratio of the distal retention zone (60%) into the above formula, we get:

[0061]

[0062] Therefore, it was determined that 72 mL of *Lactobacillus plantarum* culture would be released first from the distal retention zone, and the remaining 48 mL would be released from the proximal retention zone 3 minutes later. Through this segmented release method, the *Lactobacillus plantarum* culture does not enter the aseptic replenishment channel all at once, but rather enters the main fermentation chamber in the order of distal to proximal after passing the release check, thereby reducing the impact of the instantaneous inoculum release shock on the fermentation system.

[0063] After the *Lactobacillus plantarum* culture is added to the main fermentation chamber, the control unit can again obtain the fermentation material in the main fermentation chamber through the aseptic sampling channel and send it to the bypass test chamber for verification. When the verification result is still within the preset release range, the control unit maintains the allowable addition state of the corresponding aseptic addition channel; when the verification result is not within the preset release range, the control unit closes the corresponding aseptic addition channel and discharges the material that failed the verification in the bypass test chamber through the waste liquid sterilization channel. Through the above settings, this embodiment can verify the impact of *Lactobacillus plantarum* addition on the fermentation system online without directly interfering with the fermentation state of the main fermentation chamber, and achieve stable correction of the target strain inoculation ratio through cell retention, release, backwashing, and segmented addition.

[0064] Example 2:

[0065] This embodiment provides a method for online correction of multi-strain inoculation ratios in specialized equipment for fermented food production. This method can be executed by specialized equipment for fermented food production installed by a specific unit. The equipment includes a main fermentation chamber, a bypass test chamber, a sterile sampling channel, at least two sterile inoculation channels respectively connected to storage units of different strains, a waste liquid sterilization channel, and a control unit. The main fermentation chamber is used to contain fermentation materials. The bypass test chamber is used to perform test inoculation reactions and verification before the target strain is added to the main fermentation chamber. The sterile inoculation channels are used to add bacterial solutions of the corresponding target strains to the main fermentation chamber.

[0066] In this embodiment, the fermentation material in the main fermentation chamber is 1500L, and the fermentation material is a mixed fermentation substrate used in the production process of a certain fermented food. The control unit detects the inoculation ratio of each target strain in the main fermentation chamber through an online detection unit. The target strains include Lactobacillus plantarum, Saccharomyces cerevisiae, and Acetic Acid Bacillus. The current inoculation ratio of Lactobacillus plantarum is 18.6%, corresponding to a lower limit of 21.0% for the fermentation stage; the current inoculation ratio of Saccharomyces cerevisiae is 12.4%, corresponding to a lower limit of 14.0% for the fermentation stage; and the current inoculation ratio of Acetic Acid Bacillus is 6.8%, corresponding to a lower limit of 8.0% for the fermentation stage. The control unit determines that the inoculation ratios of the above three target strains are all lower than the corresponding lower limits, and online correction for multiple strains is required.

[0067] like Figure 5 As shown, the current inoculation rates of the three target bacterial species are all below the corresponding set lower limits. Figure 5 By comparing the current inoculation ratio with the set lower limit in parallel columns, the preconditions for the control unit to trigger multi-species correction can be intuitively mapped to each target species. Among them, Lactobacillus plantarum, Saccharomyces cerevisiae and Acetic Acid Bacillus all have a difference range from the current ratio to the set lower limit, indicating that it is not appropriate to activate the single species addition logic at this stage, but to enter the multi-target species addition judgment one by one.

[0068] When the inoculation ratios of at least two target microbial strains are both below the lower limit set for the corresponding fermentation stage, the control unit does not simultaneously open multiple aseptic replenishment channels. Instead, it first sends the fermentation material from the main fermentation chamber to the bypass replenishment chamber through the aseptic sampling channel, and then adds Lactobacillus plantarum, Saccharomyces cerevisiae, and Acetobacter acetic acid bacteria solutions to the bypass replenishment chamber for independent replenishment reactions. For each target microbial strain, the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate are collected for each replenishment reaction. A release determination result is formed based on whether the above parameters are within the preset release range.

[0069] The control unit records the time taken for each target microbial strain from the addition of the bacterial solution to the bypass test chamber until it first meets the preset release range. The time taken for *Lactobacillus plantarum* to first meet the preset release range was 18 minutes, for *Saccharomyces cerevisiae* it was 26 minutes, and for *Acetobacter acetic acid* it was 22 minutes. The control unit also determines the verification risk coefficient based on historical verification results, the current fermentation stage, and the characteristics of the target microbial strain. The verification risk coefficient for *Lactobacillus plantarum* is 0.05, for *Saccharomyces cerevisiae* it is 0.03, and for *Acetobacter acetic acid* it is 0.08. The time weighting correction factor is 8 minutes.

[0070] In this embodiment, the order of insertion is determined by the following formula:

[0071]

[0072] in, Indicates the first The determination value for the order of addition of each target bacterial species. The smaller the value, the higher the priority for filling it in; Indicates the first The time taken for a target bacterial strain from the addition of bacterial solution to the bypass test cavity until the preset release range is met for the first time; Indicates the first The calculation formula is based on the re-verification risk coefficient of each target strain; 8 represents the time weighting correction coefficient, which is used to convert the impact of re-verification risk on the replenishment order to the order of minutes. The derivation logic of this formula is that the replenishment order is mainly determined by the time when the target strain reaches the release conditions. At the same time, the impact of re-verification risk on the stability of the main fermentation chamber is considered. A risk correction term is added to the reaction time to give target strains with shorter reaction time and lower re-verification risk a higher replenishment priority.

[0073] Substituting the above data into the order determination formula one by one, we get:

[0074]

[0075]

[0076]

[0077] like Figure 6 As shown, the addition order determination value for Lactobacillus plantarum was 18.40, for Acetic Acid Bacillus it was 22.64, and for Saccharomyces cerevisiae it was 26.24. Figure 6 Presenting the initial release time and the replenishment order determination value simultaneously shows that the replenishment order is not only based on the initial release time, but also incorporates the risk correction factor for review. Since Lactobacillus plantarum has the lowest determination value, its area is identified as the priority replenishment area, and the control unit determines the replenishment order as Lactobacillus plantarum, Acetobacter, and Saccharomyces cerevisiae.

[0078] The control unit first opens the aseptic replenishment channel corresponding to *Lactobacillus plantarum*, allowing the *Lactobacillus plantarum* culture to be added to the main fermentation chamber. Before the *Lactobacillus plantarum* is fully added and a verification result is obtained, the aseptic replenishment channels corresponding to *Acetobacter* and *Saccharomyces cerevisiae* remain closed. After the *Lactobacillus plantarum* culture is added to the main fermentation chamber, the control unit obtains a sample of the material from the main fermentation chamber containing the added *Lactobacillus plantarum* through the aseptic sampling channel and sends this sample to the bypass test replenishment chamber for verification. During the verification process, the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate are still collected, and a verification result is generated according to the same preset release range as the release judgment. If the verification result is still within the preset release range, the control unit allows the next target strain to be added; if the verification result is not within the preset release range, the control unit keeps the aseptic replenishment channel corresponding to the subsequent target strain closed.

[0079] When the verification result after adding *Lactobacillus plantarum* is not within the preset release range, the control unit switches the aseptic inoculation channel corresponding to *Lactobacillus plantarum* to intermittent inoculation mode, performing low-amplitude corrections according to the preset single inoculation amount and preset inoculation interval. At this time, the target inoculation ratio of *Lactobacillus plantarum* is 21.0%, the current inoculation ratio is 18.6%, the low-amplitude correction ratio is 0.8% each time, and the preset inoculation interval is 10 minutes.

[0080] In this embodiment, the number of intermittent replenishments is determined by the following formula:

[0081]

[0082] in, Indicates the first The number of intermittent replenishment operations required for each target bacterial species; Indicates rounding up; This indicates the target inoculation ratio of the target strain at the current fermentation stage; This indicates the current inoculation percentage of the target bacterial species; This represents the low-amplitude correction ratio corresponding to each intermittent supplementation. The derivation logic of this formula is that when the review and judgment result is not within the preset release range, the system will no longer perform a one-time supplementation, but will split the difference between the target vaccination ratio and the current vaccination ratio into several low-amplitude corrections. Each correction does not exceed the preset single correction range, so it is necessary to round up the ratio of the difference to the single correction range.

[0083] Substituting the target inoculation rate of Lactobacillus plantarum (21.0%), the current inoculation rate (18.6%), and the small-amplitude correction rate (0.8%) for each inoculation into the formula for calculating the number of intermittent replenishments, we obtain:

[0084]

[0085] like Figure 7 As shown, the intermittent addition of Lactobacillus plantarum was divided into 3 additions, with an interval of 10 minutes between each addition, and the addition was followed by a checkpoint. Figure 7 The process also illustrates the control relationship between the verification results and the status of subsequent channels: when the verification after supplementation fails, the subsequent aseptic supplementation channel remains closed; when the verification passes and the process moves to the next target strain control, both Acetic Acid Bacillus and Saccharomyces cerevisiae must first undergo sequential compatibility verification, and only after the verification passes can the corresponding aseptic supplementation channel be opened. This process establishes a continuous control relationship between intermittent supplementation, post-supplementation verification, channel closure, and sequential compatibility verification.

[0086] Based on this, the control unit determines that *Lactobacillus plantarum* needs to be added three times intermittently, with a 10-minute interval between each addition, and each intermittent addition is verified by a bypass test chamber. If the verification result after any intermittent addition is not within the preset release range, the control unit stops subsequent intermittent additions and keeps the aseptic addition channels for *Acetobacter* and *Saccharomyces cerevisiae* closed; if the verification results are all within the preset release range, the control unit allows the next target strain to undergo sequence compatibility verification.

[0087] Before activating the aseptic replenishment channel for Acetic Acid Bacillus, the control unit obtains a fermentation material sample containing *Lactobacillus plantarum* that has already been added from the main fermentation chamber. This sample, along with the Acetic Acid Bacillus culture, is then added to the bypass test replenishment chamber, allowing the Acetic Acid Bacillus culture to undergo a test replenishment reaction in a material environment already containing the influence of *Lactobacillus plantarum*. The control unit generates a sequence compatibility verification result based on the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate within the bypass test replenishment chamber. If the sequence compatibility verification result is still within the preset release range, the control unit activates the aseptic replenishment channel for Acetic Acid Bacillus; otherwise, the control unit keeps the aseptic replenishment channel for Acetic Acid Bacillus closed.

[0088] After acetic acid bacteria are added to the main fermentation chamber and a verification result is obtained, the control unit performs a sequential compatibility verification on the brewing yeast in the same manner. Specifically, the control unit obtains a sample of the main fermentation chamber containing the added Lactobacillus plantarum and acetic acid bacteria, and adds this sample and the brewing yeast culture solution together to the bypass test chamber for a test reaction. The control unit only opens the aseptic inoculation channel for the brewing yeast if the sequential compatibility verification result is still within the preset release range; if the sequential compatibility verification result is not within the preset release range, the aseptic inoculation channel for the brewing yeast remains closed.

[0089] In this embodiment, when multiple target strains are below their corresponding set lower limits, the order of adding each target strain is first determined by using a bypass test chamber. Then, a verification is performed after each target strain is added. If the verification is abnormal, the system switches to intermittent addition or stops adding the strain. At the same time, a sequence compatibility verification is performed before adding the next target strain, thereby reducing the risk of secondary deviation of the inoculation ratio and fluctuation of fermentation state caused by the simultaneous addition of multiple strains.

Claims

1. A method for online correction of multi-strain inoculation ratio in specialized equipment for fermented food production, characterized in that, The equipment includes a main fermentation chamber, a bypass test chamber connected to the main fermentation chamber, a sterile sampling channel, at least two sterile replenishment channels respectively connected to different strains of liquid storage units, and a waste liquid sterilization channel. When the inoculation ratio of any target strain in the main fermentation chamber is lower than the set lower limit of the corresponding fermentation stage, the fermentation material of the preset sampling amount is sent into the bypass test chamber through the aseptic sampling channel, and the bacterial solution of the target strain is added to the bypass test chamber for test reaction. Release determination results are formed based on the pH change rate, gas production rate, redox potential change rate, and target metabolite concentration change rate in the bypass test chamber. When the release determination results are within the preset release range, the corresponding aseptic replenishment channel is opened to replenish the bacterial solution of the target strain into the main fermentation chamber. When the release determination results are not within the preset release range, the corresponding aseptic replenishment channel is closed, and the material in the bypass test chamber is discharged through the waste liquid sterilization channel.

2. The correction method according to claim 1, characterized in that, The bypass test chamber is divided into a baseline zone, a test zone, and an excess zone. Fermentation materials from the same batch, drawn through the aseptic sampling channel, enter the baseline zone, the test zone, and the excess zone, respectively. No bacterial solution of the target strain is added to the baseline zone, a first preset inoculation amount of bacterial solution of the target strain is added to the test zone, and a second preset inoculation amount of bacterial solution of the target strain is added to the excess zone. The second preset inoculation amount is greater than the first preset inoculation amount. When the release determination result of the test replenishment area is within the preset release range, and the excess area does not show a pH change rate exceeding the preset mutation threshold, a gas production rate exceeding the preset gas production upper limit, or a target metabolite concentration change direction opposite to the preset target direction, the corresponding aseptic replenishment channel is opened.

3. The correction method according to claim 1, characterized in that, The bypass test chamber is equipped with a bacterial cell retention component, which allows soluble substrates and metabolites to pass through, while blocking the target bacterial species from entering the sterile bacterial replenishment channel from the bypass test chamber. The bacterial solution of the target strain first comes into contact with the fermentation material in the bacterial retention component. When the release determination result is within the preset release range, the corresponding aseptic replenishment channel is opened. When the release determination result is not within the preset release range, the material in the bacterial retention component is discharged into the waste liquid sterilization channel.

4. The correction method according to claim 1, characterized in that, When the inoculation ratio of at least two target strains is lower than the set lower limit of the corresponding fermentation stage, the aseptic replenishment channel is opened one by one according to the bypass replenishment judgment result; wherein, before the bacterial solution of one target strain is replenished into the main fermentation chamber and verified by the bypass replenishment chamber, the aseptic replenishment channel corresponding to other target strains remains closed.

5. The correction method according to claim 3, characterized in that, The cell retention component has a retention state and a release state; in the retention state, the target bacterial cells are confined within the cell retention component, and the exchange of soluble substrates and metabolites within the bypass test chamber is allowed. When the release determination result is within the preset release range, the bacterial cell retention component switches to the release state, so that the retained target bacterial cells enter the sterile replenishment channel along with the corresponding target bacterial solution.

6. The correction method according to claim 3, characterized in that, The bacterial cell retention component is connected to a backwash branch; when the release determination result is not within the preset release range, the corresponding aseptic replenishment channel is closed first, and then the target bacterial cells and residual fermentation materials in the bacterial cell retention component are pushed to the waste liquid sterilization channel through the backwash branch.

7. The correction method according to claim 3, characterized in that, The bacterial retention component is divided into a proximal retention zone and a distal retention zone in the same test reaction. The proximal retention zone is located upstream of the direction in which the target bacterial culture enters, and the distal retention zone is located downstream of the direction in which the target bacterial culture enters. The target bacterial culture enters the proximal retention zone first. When the release determination result is within the preset release range, the distal retention zone is released first, and the proximal retention zone is released after a preset delay time.

8. The correction method according to claim 4, characterized in that, First, open the aseptic inoculation channel corresponding to the target bacterial strain in the bypass test chamber that takes the shortest time from the addition of the target bacterial culture to the first time the preset release range is met. After the bacterial culture of the target strain is added to the main fermentation chamber, if the verification result of the bypass test chamber is still within the preset release range, the aseptic replenishment channel corresponding to the next target strain will be opened.

9. The correction method according to claim 4, characterized in that, After the bacterial culture of the current target strain is added to the main fermentation chamber, if the verification result of the bypass test chamber is not within the preset release range, the aseptic replenishment channel corresponding to the subsequent target strain remains closed, and the aseptic replenishment channel corresponding to the current target strain is switched to an intermittent replenishment state that is executed according to a preset single replenishment amount and a preset replenishment interval, or switched to a stopped replenishment state.

10. The correction method according to claim 8, characterized in that, Before the aseptic replenishment channel corresponding to the next target strain is opened, the material sample of the main fermentation chamber containing the target strain and the bacterial solution of the next target strain are added together to the bypass test replenishment chamber for sequential compatibility verification. When the verification result of the bypass test cavity is still within the preset release range, the aseptic replenishment channel corresponding to the next target bacterial species is opened; when the verification result of the bypass test cavity is not within the preset release range, the aseptic replenishment channel corresponding to the next target bacterial species remains closed.

Images