A composite microbial agent culture device and method

Abstract

The application discloses a kind of compound microbial inoculant culture device and method, and culture device includes culture tank and adjusting mechanism.The top of culture tank is equipped with feeding port and air vent, and the bottom is equipped with discharge port;Adjusting mechanism includes adjusting plate, clamping plate, rectangular frame and moving block, clamping plate is equipped with adjusting plate slot, rectangular frame slot and moving block slot in sequence, adjusting plate is located in adjusting plate slot, rectangular frame is located in rectangular frame slot, and moving block is located in moving block slot;The bottom of adjusting plate is inclined plane, and the outer wall of roller is tightly attached to inclined plane and the top of moving block simultaneously, the left wall of rectangular frame is connected to clamping plate by first spring, the right wall of rectangular frame is equipped with air vent conversion plate, the top of moving block is equipped with horizontal area and inclined area, the bottom of moving block is equipped with push rod, and push rod is connected to sealing plate after penetrating the bottom of clamping plate, and the other end of sealing plate is located at discharge port.It can be simultaneously mixed and symbiotic culture for lactic acid bacteria, photosynthetic bacteria, yeast and nitrogen-fixing bacteria, etc., and a large number of proliferation and keep strain activity.

Description

Technical Field

[0001] This invention relates to the field of compound microbial agent cultivation technology, and in particular to a compound microbial agent cultivation device and method. Background Technology

[0002] Microbial inoculants are commonly used and highly effective green ecological products in agricultural soil improvement, ecological environment management, and aquaculture efficiency enhancement. Among them, photosynthetic bacteria are one of the important bacterial species. However, single photosynthetic bacteria inoculants are not easy to colonize and survive in the application environment in practical applications, making it difficult to achieve the expected results. Compound inoculants, due to the complexity of strain selection, strain compatibility, culture medium selection, and culture conditions, often exhibit unstable population structure, weak synergistic effects between strains, and even mutual antagonism, competition, and inhibition. They cannot coexist and thrive, nor can they form a stable micro-ecosystem, thus failing to achieve the expected goals. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention provides a compound microbial agent cultivation device and method that enables the simultaneous mixed symbiotic cultivation of bacteria such as lactic acid bacteria, photosynthetic bacteria, yeast, and nitrogen-fixing bacteria, resulting in massive proliferation and maintenance of strain activity. After application, these bacteria can colonize and promote growth in the environment, working synergistically to form a stable and long-lasting ecosystem.

[0004] In order to achieve the objective of this invention, the following solution is proposed:

[0005] A compound microbial agent culture device is used to culture one of photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria and yeast. The culture device includes a culture tank and a regulating mechanism.

[0006] The culture tank is equipped with a feeding port and a vent at the top, and a discharge port at the bottom.

[0007] The adjustment mechanism is located on the left side of the culture tank. The adjustment mechanism includes an adjustment plate, a retaining plate, a rectangular frame, and a moving block. The retaining plate has an adjustment plate slot, a rectangular frame slot, and a moving block slot from top to bottom. The adjustment plate slides up and down to engage with the adjustment plate slot, the rectangular frame slides left and right to engage with the rectangular frame slot, and the moving block slides up and down to engage with the moving block slot.

[0008] The bottom of the adjusting plate is an inclined surface facing the culture tank. Rollers are installed inside the rectangular frame. The front and rear sides of the rollers are connected to the inner wall of the rectangular frame through bearings. The outer wall of the rollers is in close contact with the inclined surface and the top of the moving block. The left wall of the rectangular frame is connected to the left wall of the clamping plate through a first spring. The right wall of the rectangular frame is equipped with a venting conversion plate. The venting conversion plate moves left and right to open or close the vent. From left to right, the top of the moving block has a horizontal area and an upward inclined area. The bottom of the moving block is equipped with a push rod. The push rod passes through the bottom of the clamping plate and is connected to a sealing plate. A second spring is installed outside the push rod. The two ends of the second spring are connected to the moving block and the clamping plate. The sealing plate is set horizontally. The other end of the sealing plate is located at the discharge port. The sealing plate moves up and down to open or close the discharge port.

[0009] Furthermore, from right to left, the ventilation conversion plate is provided with a ventilation zone, a sealing zone, and a ventilation area. The ventilation zone has several ventilation holes. When the roller is located at the junction of the horizontal zone and the inclined zone, the sealing zone closes the ventilation holes.

[0010] Furthermore, the top of the culture tank is equipped with two sealing strips, and a sealing plate is located between the two sealing strips.

[0011] Furthermore, when the roller is located at the junction of the horizontal and inclined areas, the first spring is in a compressed state, the second spring is in a natural or compressed state, and the discharge port is in a closed state.

[0012] Furthermore, the top of the adjustment plate is equipped with a lifting rod, and the top of the adjustment plate is equipped with lugs on both sides. The bottom of the lugs is connected to the card plate by a third spring.

[0013] Furthermore, the front wall of the adjusting plate is provided with a through groove, and the rear wall of the adjusting plate groove is provided with a guide rod, which slides up and down within the through groove.

[0014] A method for cultivating compound microbial agents includes the following steps:

[0015] Culture of bacterial solution: Photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria and yeast were cultured using four of the above-described compound bacterial agent culture devices, and the bacterial solutions were prepared by propagation;

[0016] Cultivating compound microbial agents: Using PLA medium, under aseptic conditions, inoculate with lactic acid bacteria at an initial concentration of 0.5~1.2·10⁻⁶. 5 CFU / ml, inoculum size 8-15 ml / L, static culture for 3-6 days, adjust fermentation broth pH to 7.5-8 with NaOH solution, inoculate under aseptic conditions with photosynthetic bacteria, nitrogen-fixing bacteria, and yeast cultures, all with an initial concentration of 0.5-1.2 × 10⁻⁶. 5 CFU / ml, in a ratio of 10:5:8 ml / L, at 20~40℃, 2500~3500 Lux, and incubated statically for 8~15 days.

[0017] Furthermore, the PLA culture medium consists of: yeast extract 1.0~2.5g, mannitol 1.0~3.0g, glucose 1.0~2g, TES stock solution 0.5~1.5ml, distilled water 800~1200ml, pH 7.0~7.5, sterilized at 121℃ for 15min.

[0018] Furthermore, the specific steps for culturing the bacterial solution are as follows:

[0019] Add one strain of photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria, or yeast, along with the ingredients and distilled water, into the culture tank through the feeding port. Seal the feeding port. At this point, the roller is located at the junction of the horizontal and inclined zones.

[0020] After a period of cultivation, the adjustment plate begins to move upward, and the rectangular frame, subjected to the leftward elastic force of the first spring, begins to move to the left, causing the air vent to shift onto the air vent.

[0021] After oxygenation is completed, the adjustment plate moves down to its original position, and the rollers are squeezed by the inclined plane, causing the rectangular frame to move to the right to its original position.

[0022] After a period of further cultivation, the regulating plate continues to move downwards, and the rollers are squeezed by the inclined plane, causing the rectangular frame to continue to move to the right. Another ventilation zone moves to the ventilation port. At the same time, the rollers squeeze the inclined area, causing the moving block and sealing plate to begin to move downwards. The sealing plate separates from the discharge port and begins to release the bacterial liquid.

[0023] The beneficial effects of this invention are as follows:

[0024] 1. Culture Method: Using photosynthetic bacteria as the core strain, compatible strains of nitrogen-fixing bacteria, lactic acid bacteria, and yeast were screened. Through a special culture formula and process, all four strains could grow and proliferate in the same culture medium, achieving a cell concentration of 102. 8 With a concentration of CFU / ml or higher, and a stable community structure for more than 30 days while maintaining vitality, the plant can colonize and promote growth in the environment after application, working synergistically to form a stable and long-lasting ecosystem, thereby achieving the goals of improving soil quality, restoring the ecosystem, and increasing yield and quality.

[0025] 2. Cultivation device: The structure is very ingenious. Through the cooperation of the adjustment plate, the clamping plate, the rectangular frame and the moving block, the vent of the cultivation tank can be opened and sealed by simply moving the adjustment plate up and down. The discharge port can also be opened and sealed. Attached Figure Description

[0026] Figure 1 This is a structural diagram of the culture device for an embodiment;

[0027] Figure 2 This is a plan perspective view of the culture apparatus in the embodiment;

[0028] Figure 3 This is a diagram of the cardboard structure for an embodiment;

[0029] Figure 4 This is a structural diagram of the adjustment plate in an embodiment;

[0030] Figure 5 This is a rectangular frame structure diagram of an embodiment;

[0031] Figure 6 This is a structural diagram of the moving block in an embodiment;

[0032] Figure 7 The following are structural diagrams of four culture devices used in the embodiment;

[0033] Figure 8 This is a flowchart of the cultivation method for an example. Detailed Implementation

[0034] like Figures 1-8 As shown, this embodiment provides a compound microbial agent culture device, which can culture one of photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria and yeast. The culture device includes a culture tank 1 and an adjustment mechanism 2.

[0035] Specifically, culture tank 1 is mounted on a support frame, such as... Figure 2 As shown, the top of the culture tank 1 is provided with a feeding port 11 and a vent 12, and the bottom is provided with a discharge port 13. The culture tank 1 is also provided with a light control system, a temperature control system, and a humidity control system, which are used to regulate the light, temperature and humidity inside the culture tank 1, respectively.

[0036] Specifically, the adjustment mechanism 2 is located on the left side of the culture tank 1. The adjustment mechanism 2 includes an adjustment plate 21, a locking plate 22, a rectangular frame 23, and a moving block 24, as shown below. Figure 3 As shown, the card plate 22 is vertically arranged and fixed on the support frame. From top to bottom, it is provided with an adjustment plate groove 221, a rectangular frame groove 222 and a moving block groove 223 that are interconnected. The adjustment plate 21 slides up and down in the adjustment plate groove 221. The top and bottom surfaces of the rectangular frame 23 are connected. The rectangular frame 23 slides left and right in the rectangular frame groove 222. The moving block 24 slides up and down in the moving block groove 223.

[0037] Specifically, such as Figure 4 As shown, the bottom of the adjusting plate 21 is an inclined surface 211 facing the culture tank 1. The top of the adjusting plate 21 is provided with a lifting rod 214. The power source of the lifting rod 214 can be a cylinder or the like. The top two sides of the adjusting plate 21 are provided with lugs 212. The bottom of the lugs 212 is connected to the clamping plate 22 by a third spring 213. The front wall of the adjusting plate 21 is provided with a through groove 215. The rear wall of the adjusting plate groove 221 is provided with a guide rod 2211. The guide rod 2211 slides up and down in the through groove 215.

[0038] Specifically, such as Figure 5 As shown, a roller 231 is provided inside the rectangular frame 23. The front and rear sides of the roller 231 are connected to the inner wall of the rectangular frame 23 through bearings. The roller 231 can rotate around the bearings. The outer wall of the roller 231 is tangent to the inclined surface 211 and the top of the moving block 24. The left wall of the rectangular frame 23 extends downwards for a section. The extended section is connected to the left wall of the card plate 22 through the first spring 25.

[0039] Specifically, the right wall of the rectangular frame 23 is provided with a ventilating conversion plate 233. The ventilating conversion plate 233 is located at the top of the culture tank 1. From right to left, the ventilating conversion plate 233 is provided with a ventilating area 2331 and a sealing area 2332. The ventilating area 2331 is provided with several vent holes. The ventilating conversion plate 233 can move left and right, and the ventilating area 2331 and the sealing area 2332 can alternately cover the vent 12, thereby opening or closing the vent 12.

[0040] Specifically, such as Figure 6 As shown, from left to right, the top of the moving block 24 is provided with a horizontal area 241 and an upwardly inclined area 242. The angle between the top surface of the horizontal area 241 and the top surface of the inclined area 242 is an obtuse angle. The bottom of the moving block 24 is provided with a push rod 243. The push rod 243 passes through the bottom of the clamping plate 22 and is connected to a sealing plate 26. A second spring 244 is provided outside the push rod 243. The two ends of the second spring 244 are connected to the moving block 24 and the clamping plate 22. The sealing plate 26 is set horizontally. The other end of the sealing plate 26 is located at the discharge port 13. The sealing plate 26 moves up and down to open or close the discharge port 13.

[0041] Specifically, in order to improve the sealing performance, the top of the culture tank 1 is provided with two sealing strips 14, and the sealing plate 26 is located between the two sealing strips 14.

[0042] In this embodiment, in the initial state, the roller 231 is located at the junction of the horizontal area 241 and the inclined area 242, the first spring 25 is in a compressed state, the second spring 244 is in a natural or compressed state, the discharge port 13 is just in a closed state, and the sealing area 2332 just closes the vent 12.

[0043] If the lifting rod 214 rises, the adjusting plate 21 begins to move upward, and the roller 231 loses the pressure of the inclined plane 211. Because the rectangular frame 23 is still subjected to the leftward elastic force of the first spring 25, the rectangular frame 23 and the roller 231 begin to move to the left, thereby causing the ventilation area 2331 to shift onto the ventilation port 12. During this process, the roller 231 always moves in the horizontal area 241, so the moving block 24 remains stationary.

[0044] If, starting from the initial state, the lifting rod 214 descends, the adjusting plate 21 moves downward, and the roller 231 is squeezed by the inclined plane 211, causing the rectangular frame 23 to move to the right, then another venting area 2331 will move to the vent 12. At the same time, the roller 231 squeezes the inclined area 242, causing the moving block 24 and the sealing plate 26 to begin to move downward, and the sealing plate 26 disengages from the discharge port 13.

[0045] Example 2

[0046] like Figure 8 As shown, this embodiment provides a method for cultivating compound microbial agents, including the following steps:

[0047] Culture medium: such as Figure 7 As shown, photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria and yeast were cultured using the compound bacterial agent culture device in four Examples 1, and bacterial solutions were prepared by propagation.

[0048] The specific operation is as follows: one strain of photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria and yeast, ingredients and distilled water are put into the culture tank 1 through the feeding port 11 and the feeding port 11 is closed. At this time, the roller 231 is located at the junction of the horizontal area 241 and the inclined area 242. During the culture of bacterial solution, it is mixed evenly. It is also necessary to adjust the appropriate parameters such as temperature, brightness and humidity, and set the culture time.

[0049] After a period of cultivation, oxygen needs to be introduced into the culture tank 1. The regulating plate 21 begins to move upward, and the rectangular frame 23 is subjected to the leftward elastic force of the first spring 25, and begins to move to the left, so that the venting area 2331 is moved to the venting port 12.

[0050] After oxygenation is completed, the adjusting plate 21 moves downward to its original position, and the roller 231 is squeezed by the inclined surface 211, causing the rectangular frame 23 to move to the right to its original position.

[0051] After a period of further cultivation, the adjusting plate 21 continues to move downwards, and the roller 231 is squeezed by the inclined surface 211, causing the rectangular frame 23 to continue to move to the right. Another venting area 2331 moves to the vent 12. At the same time, the roller 231 squeezes the inclined area 242, causing the moving block 24 and the sealing plate 26 to begin to move downwards. The sealing plate 26 separates from the discharge port 13 and begins to release the bacterial liquid.

[0052] More specifically, when culturing photosynthetic bacteria, you can add 0.5g of dipotassium hydrogen phosphate, 0.5g of potassium dihydrogen phosphate, 1g of ammonium sulfate, 0.5g of magnesium sulfate, 1g of sodium acetate, 0.2g of yeast powder, 1g of beef extract, and 1000ml of distilled water. Cultivate at 30℃ and 3000lx light for a total of 10 days, and then gradually expand the bacteria at an inoculum of 10%.

[0053] When culturing nitrogen-fixing bacteria, add 1.0g yeast extract, 10.0g mannitol, 1ml TES, 15.0g agar, 800ml distilled water, pH 7.2, 30℃, and culture for 2 days. Then, gradually expand the culture at an inoculum of 5%.

[0054] When culturing lactic acid bacteria, you can add 10.0g of peptone, 5.0g of beef meal, 4.0g of yeast powder, 20.0g of glucose, 5.0g of sodium acetate, 2.0g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 2.0g of triammonium citrate, 0.05g of manganese sulfate, and 1ml of Tween 80. Incubate at 34℃ for 2 days, and then gradually propagate at an inoculum of 5%.

[0055] When culturing yeast, you can add 1% yeast extract, 2% peptone, and 2% glucose, and culture at 26°C for a total of 2 days. Then, gradually expand the culture by inoculating at a rate of 5%.

[0056] It should be noted that the culture parameters of the compound microbial agents listed above are not unique and fixed, and can fluctuate by 0.5% or 1% as the upper and lower limits.

[0057] After the bacterial culture was completed, the compound bacterial agent was cultured: PLA medium was used, and under aseptic conditions, the lactic acid bacteria culture was inoculated at an initial concentration of 0.5~1.2·10⁻⁶. 5 CFU / ml, inoculum size 8-15 ml / L, static culture for 3-6 days, adjust fermentation broth pH to 7.5-8 with NaOH solution, inoculate under aseptic conditions with photosynthetic bacteria, nitrogen-fixing bacteria, and yeast cultures, all with an initial concentration of 0.5-1.2 × 10⁻⁶. 5 The compound bacterial agent was obtained by incubating at 20-40℃ and 2500-3500 Lux for 8-15 days with cfu / ml in a ratio of 10:5:8 ml / L, and then incubating for 8-15 days.

[0058] Preferably, PLA medium is used, and lactic acid bacteria culture is inoculated under aseptic conditions with an initial concentration of 10. 5 CFU / ml, inoculum size 10 ml / L, static culture for 5 days, adjust fermentation broth pH to 7.5-8 with NaOH solution, and inoculate under aseptic conditions with photosynthetic bacteria, nitrogen-fixing bacteria, and yeast cultures, all with an initial concentration of 10. 5 The compound bacterial agent was obtained by incubating at 30℃ and 3000 Lux for 10 days with cfu / ml in a ratio of 10:5:8 ml / L.

[0059] The PLA culture medium consisted of 1.0-2.5g yeast extract, 1.0-3.0g mannitol, 1.0-2g glucose, 0.5-1.5ml TES stock solution, 800-1200ml distilled water, pH 7.0-7.5, and sterilized at 121℃ for 15min.

[0060] Preferred PLA culture medium: 1.5g yeast extract, 2.0g mannitol, 1.5g glucose, 1ml TES stock solution, 1000ml distilled water, pH 7.2, sterilized at 121℃ for 15min.

[0061] The above embodiments are only used to illustrate the technical concept and features of the present invention, and are not intended to be unique or to limit the present invention. Those skilled in the art should understand that various changes or equivalent substitutions made to the present invention without departing from its scope are all within the protection scope of the present invention.

Claims

1. A compound microbial agent cultivation device, characterized in that, It includes a culture tank (1) and a regulating mechanism (2) for culturing one of photosynthetic bacteria, nitrogen-fixing bacteria, lactic acid bacteria and yeast; The culture tank (1) has a feeding port (11) and a vent (12) at the top and a discharge port (13) at the bottom. The adjustment mechanism (2) is located on the left side of the culture tank (1). The adjustment mechanism (2) includes an adjustment plate (21), a clamping plate (22), a rectangular frame (23), and a moving block (24). The clamping plate (22) is provided with an adjustment plate groove (221), a rectangular frame groove (222), and a moving block groove (223) from top to bottom. The adjustment plate (21) slides up and down in the adjustment plate groove (221), the rectangular frame (23) slides left and right in the rectangular frame groove (222), and the moving block (24) slides up and down in the moving block groove (223). The bottom of the adjusting plate (21) is an inclined surface (211) facing the culture tank (1). A roller (231) is provided inside the rectangular frame (23). The front and rear sides of the roller (231) are connected to the inner wall of the rectangular frame (23) via bearings. The outer wall of the roller (231) is simultaneously in close contact with the inclined surface (211) and the top of the moving block (24). The left wall of the rectangular frame (23) is connected to the left wall of the clamping plate (22) via a first spring (25). The right wall of the rectangular frame (23) is provided with a venting conversion plate (233). The venting conversion plate (233) moves left and right to open or close the vent (12). From left to right... In terms of direction, the top of the moving block (24) is provided with a horizontal area (241) and an upward inclined area (242). The bottom of the moving block (24) is provided with a push rod (243). The push rod (243) passes through the bottom of the card plate (22) and is connected to a sealing plate (26). A second spring (244) is provided outside the push rod (243). The two ends of the second spring (244) are connected to the moving block (24) and the card plate (22). The sealing plate (26) is set horizontally. The other end of the sealing plate (26) is located at the discharge port (13). The sealing plate (26) moves up and down to open or close the discharge port (13).

2. The compound microbial agent cultivation device according to claim 1, characterized in that, From right to left, the ventilation conversion plate (233) is provided with a ventilation area (2331), a sealing area (2332), and a ventilation area (2331). The ventilation area (2331) is provided with several ventilation holes. When the roller (231) is located at the junction of the horizontal area (241) and the inclined area (242), the sealing area (2332) closes the ventilation hole (12).

3. The compound microbial agent cultivation device according to claim 1, characterized in that, When the roller (231) is located at the junction of the horizontal area (241) and the inclined area (242), the first spring (25) is in a compressed state, the second spring (244) is in a natural state or a compressed state, and the discharge port (13) is in a closed state.

4. The compound microbial agent cultivation device according to any one of claims 1 to 3, characterized in that, The top of the adjustment plate (21) is provided with a lifting rod (214), and the top sides of the adjustment plate (21) are provided with lugs (212). The bottom of the lugs (212) is connected to the card plate (22) by a third spring (213).

5. The compound microbial agent cultivation device according to claim 4, characterized in that, The front wall of the adjusting plate (21) is provided with a through groove (215), and the rear wall of the adjusting plate groove (221) is provided with a guide rod (2211). The guide rod (2211) slides up and down in the through groove (215).

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